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"The corrected version of"} +{"text": "This has now been corrected online."} +{"text": "The images for Fig In In"} +{"text": "In Zuurbier et al,"} +{"text": "The images for Figs"} +{"text": "The images for Figs"} +{"text": "The original article has been updated."} +{"text": "In Nalairndran et al,"} +{"text": "The correct Supplementary Information now accompanies the Article."} +{"text": "Following publication of the original article the auth"} +{"text": "The image for"} +{"text": "The original article containsThe correct presentation of Fig."} +{"text": "The image for"} +{"text": "The images for Figs"} +{"text": "The image for"} +{"text": "The original article mistaken"} +{"text": "The original article has been updated."} +{"text": "The Editor is retracting this article because"} +{"text": "The images for Figs"} +{"text": "The images for Figs"} +{"text": "The image for"} +{"text": "The original article has been updated."} +{"text": "The original article has been updated."} +{"text": "They thank the Editor of"} +{"text": "This article has been corrected."} +{"text": "The authors have retracted this article for lega"} +{"text": "The corrected version of"} +{"text": "The images for Figs"} +{"text": "In the original article,"} +{"text": "There are formatting errors in Tables"} +{"text": "These have now been added."} +{"text": "In Lei et al,"} +{"text": "The paper by Santucci et al. gave us"} +{"text": "The caption for"} +{"text": "In Shi et al,"} +{"text": "The revised version of"} +{"text": "We appreciate Zins and Abraham commenti"} +{"text": "In Hou et al,"} +{"text": "In Lina Xuan et al,"} +{"text": "The images for Figs"} +{"text": "The image for"} +{"text": "The images for Figs"} +{"text": "We present the spinal atomic"} +{"text": "The article has been corrected online ("} +{"text": "The images for Figs"} +{"text": "The captions for Figs"} +{"text": "The images for Figs"} +{"text": "The original article has been updated."} +{"text": "The bulleted item should read:"} +{"text": "In Zhi et al,"} +{"text": "The original article has been updated."} +{"text": "In Li et al,"} +{"text": "The use"} +{"text": "We extend the"} +{"text": "In Xiong et al,"} +{"text": "The revised version of"} +{"text": "In Li et al,"} +{"text": "In Xue W et al.,"} +{"text": "The original article has been corrected."} +{"text": "The revised version of"} +{"text": "The original article has been corrected."} +{"text": "The original article has been updated."} +{"text": "The original article has been corrected."} +{"text": "The original article has been corrected."} +{"text": "The original article has been corrected."} +{"text": "The original article has been corrected."} +{"text": "In Wang H et al,"} +{"text": "The captions for Figs"} +{"text": "The original article has been updated."} +{"text": "In Li Yarui et al.,"} +{"text": "In Ying Luo et al.,"} +{"text": "The original article has been updated."} +{"text": "In Chi et al.,"} +{"text": "In Zhonghui Chen et al.,"} +{"text": "The original article has been corrected."} +{"text": "The corrected version of"} +{"text": "The corrected version of"} +{"text": "This article has been corrected."} +{"text": "The PDF has been corrected."} +{"text": "The corrected version of"} +{"text": "The original article has been corrected."} +{"text": "The original article has been updated."} +{"text": "The original article has been updated."} +{"text": "This Perspective was published withan incorrect absoluteconfiguration of compounds"} +{"text": "Details ofthe correction onlyto the Acknowledgments:"} +{"text": "The original article has been corrected."} +{"text": "The original article has been updated."} +{"text": "PLOS ONE has withdrawn this article [ article and remo"} +{"text": "The original article has been updated."} +{"text": "The original article has been corrected."} +{"text": "The revised version of"} +{"text": "The original article has been corrected.\u201d"} +{"text": "In The correct table is shown below."} +{"text": "The revised version of"} +{"text": "In Hu et al.,"} +{"text": "The correct link is:"} +{"text": "The original article has been corrected."} +{"text": "The original article has been corrected."} +{"text": "In Tang et al.,"} +{"text": "In Bofang Zhang et al.,"} +{"text": "The original article has been corrected."} +{"text": "The original article has been updated."} +{"text": "The author Replies:,,,In this issue Tsuboi and BertramS2"} +{"text": "The original article has been updated."} +{"text": "The authors agree wi"} +{"text": "The original article has been corrected.Supplementary information"} +{"text": "The original article has been corrected."} +{"text": "The original article has been corrected."} +{"text": "The original article has been corrected."} +{"text": "The original article has been corrected."} +{"text": "The original publication has also been updated."} +{"text": "The original article contains"} +{"text": "The original article has been corrected.\u201c"} +{"text": "In Song X et al.,"} +{"text": "The original article has been updated."} +{"text": "When I invited Drs Cesar and Margaret Boguszewski as gues"} +{"text": "The original article has been updated."} +{"text": "The article has been corrected online ("} +{"text": "Consult PDF for full article"} +{"text": "The article has also been corrected online ("} +{"text": "The legends for"} +{"text": "This additional information is outlined here:"} +{"text": "After publication of the original article it came"} +{"text": "The corrections are outlined here:"} +{"text": "The article has been corrected online ("} +{"text": "Abstract not submitted for online publication"} +{"text": "Please see the corrected"} +{"text": "The article by Aktipis and Nesse containe"} +{"text": "Abstract not submitted for online publication"} +{"text": "Abstract not submitted for online publication"} +{"text": "Following the publication of this work we becam"} +{"text": "The legends for"} +{"text": "Ian Lipkin. The article has been corrected online"} +{"text": "Abstract not submitted for online publication"} +{"text": "Abstract not submitted for online publication"} +{"text": "Abstract not submitted for online publication"} +{"text": "We congratulate Mensa et al."} +{"text": "Abstract not submitted for online publication"} +{"text": "The article has been corrected online ("} +{"text": "The correct link is:"} +{"text": "The editors regretfully retract the article by Jabba"} +{"text": "The caption of"} +{"text": "Abstract not submitted for publication"} +{"text": "In panel C of"} +{"text": "A mistake regarding the exemplary weights given in Table"} +{"text": "Our pet al. ."} +{"text": "After publication of the original article it came"} +{"text": "Abstract not submitted for online publication"} +{"text": "The article has been corrected online ("} +{"text": "Abstract not submitted for publication"} +{"text": "The correct email addressis:"} +{"text": "Abstract not submitted for online publication"} +{"text": "The correct file can be viewed here:"} +{"text": "After publication of this work we notic"} +{"text": "Please see the corrected"} +{"text": "The article has been corrected online ("} +{"text": "After publication of the original article it came"} +{"text": "The authors have detected a typographical error in"} +{"text": "Chemical retention calculations have been corrected in Table"} +{"text": "Abstract not submitted for publication"} +{"text": "The aut"} +{"text": "An error occurred in"} +{"text": "Abstract not submitted for online publication"} +{"text": "Please consider the following changes in"} +{"text": "Answer to rhythm puzzle"} +{"text": "Please see the correct References section here:"} +{"text": "The publisher has retracted this article because"} +{"text": "Abstract not submitted for publication"} +{"text": "Please view the corrected table here:"} +{"text": "Abstract not submitted for online publication"} +{"text": "The Figure"} +{"text": "The correct text can be viewed here:"} +{"text": "Dear Sir:We thank Cohen and others"} +{"text": "Abstract not submitted for online publication"} +{"text": "Abstract not submitted for online publication"} +{"text": "The legend for"} +{"text": "Abstract not submitted for publication"} +{"text": "The UCM"} +{"text": "Following publication of this article it came"} +{"text": "The mai"} +{"text": "The article has been corrected online ("} +{"text": "The regulation of I"} +{"text": "The corrected text can be found here:"} +{"text": "The legend for"} +{"text": "Abstract not submitted for online publication"} +{"text": "Abstract not submitted for online publication"} +{"text": "Abstract not submitted for online publication"} +{"text": "The revised manuscript can be viewed here:"} +{"text": "Abstract not submitted for online publication"} +{"text": "This General Commentary provides a corrected version of Figure"} +{"text": "Abstract not submitted for online publication"} +{"text": "During the proofing 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other by halogen bonds.The title compound was synthesized by a new type of reaction using Mg2 as a new catalyst and a possible mechanism for this reaction is proposed. The six-membered ring adopts a half-chair conformation. In the crystal, hydrogen bonds connect the mol\u00adecules into double layers, which are connected to each other by halogen bonds. The Hirshfeld surface analysis revealed that the most important contributions for the crystal packing are from O\u22efH/H\u22efO (35.8%), Cl\u22efCl (19.6%), Cl\u22efH/H\u22efCl (17.0%), H\u22efH (8.3%), C\u22efO/O\u22efC (4.3%), Cl\u22efO/O\u22efCl (4.2%) and O\u22efO (4.1%) contacts.The mol\u00adecular and crystal structures of the title compound, C The reaction scheme is shown in Fig.\u00a015,5-Di\u00adchloro-6-hy\u00addroxy\u00addihydro\u00adpyrimidine-2,4\u2005\u00c5 x, y\u00a0+\u00a01, z; (ii) x, \u2212y\u00a0+\u00a02, z\u00a0+\u00a0A\u22efO1i hydrogen bond also involves O1i with a H4A\u22efO1i distance of 3.058\u2005(2)\u2005\u00c5 \u2212x, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01; (v) x, \u2212y\u00a0+\u00a02, z\u00a0\u2212\u00a0A\u22efO2iii with an N1\u22efO2iii distance of 2.793\u2005(2)\u2005\u00c5. The hydrogen bonds connect the mol\u00adecules into double layers parallel to the (100) plane, as shown in Fig.\u00a04vi [3.3670\u2005(9)\u2005\u00c5] and Cl2\u22efCl2vii connect the layers, forming a three-dimensional framework.The hydrogen-bond system is shown in Fig.\u00a03\u00c5 Table\u00a02. In the Crystal Explorer 17.5 to 0.986 (blue) a.u., as illustrated in Fig.\u00a05dnorm plot indicate inter\u00admolecular contacts involving the hydrogen and halogen bonds. The brightest red spots correspond to the strongest hydrogen bonds, N1\u2014H1A\u22efO2 and O3\u2014H3\u22efO2 until the components were completely dissolved. Crystallization occurred with isothermal evaporation of the resulting solution at room temperature for several weeks, giving colourless needle-shaped crystals, composition according to chemical analysis : C, 24.12/24.14; H, 2.04/2.03; Cl, 35.64/35.63; N, 14.07/14.08; O, 24.13/24.12. Crystals suitable for a X-ray structural analysis were extracted manually from this batch.The title compound was synthesized by adding 5\u2005mg of uracil (Sigma Aldrich) to 1\u2005ml of 1\u2005mol\u2005l4\u2212 does not react with HCl while TcO4\u2212 is actively reduced and possibly through radical substitution.We suggest a possible mechanism of the observed reaction. Typically, ReOUiso(H) = 1.2Ueq(C)]. O- and N-bound H atoms were refined isotropically [Uiso(H) = 1.2Ueq].Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989020011809/zq2257sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020011809/zq2257Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020011809/zq2257Isup3.cmlSupporting information file. DOI: 2025758CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A linear gold-atom geometry defined by phosphane-P and thiol\u00adate-S atoms is found in the title compound. The packing is stabilized by a combination of fluoro\u00adbenzene-C\u2014H\u22efO(meth\u00adoxy), phenyl-C\u2014H\u22efF, phenyl-C\u2014H\u22efS(thiol\u00adate) and phenyl-C\u2014H\u22ef\u03c0 inter\u00adactions to generate a three-dimensional network. 26H22AuFNOPS or [Au(C8H7FNOS)(C18H15P)], has the AuI centre coordinated by phosphane-P [2.2494\u2005(8)\u2005\u00c5] and thiol\u00adate-S [2.3007\u2005(8)\u2005\u00c5] atoms to define a close to linear geometry [P\u2014Au\u2014S = 176.10\u2005(3)\u00b0]. The thiol\u00adate ligand is orientated so that the meth\u00adoxy-O atom is directed towards the Au atom, forming an Au\u22efO close contact of 2.986\u2005(2)\u2005\u00c5. In the crystal, a variety of inter\u00admolecular contacts are discerned with fluoro\u00adbenzene-C\u2014H\u22efO(meth\u00adoxy) and phenyl-C\u2014H\u22efF inter\u00adactions leading to dimeric aggregates. These are assembled into a three-dimensional architecture by phenyl-C\u2014H\u22efS(thiol\u00adate) and phenyl-C\u2014H\u22ef\u03c0 inter\u00adactions. Accordingly, the analysis of the calculated Hirshfeld surface shows 30.8% of all contacts are of the type C\u22efH/H\u22efC but this is less than the H\u22efH contacts, at 44.9%. Other significant contributions to the surface come from H\u22efF/F\u22efH [8.1%], H\u22efS/S\u22efH [6.9%] and H\u22efO/O\u22efH [3.2%] contacts. Two major stabilization energies have contributions from the phenyl-C\u2014H\u22ef\u03c0(fluoro\u00adbenzene) and fluoro\u00adbenzene-C\u2014H\u22efC(imine) inter\u00adactions , and from the fluoro\u00adbenzene-C\u2014H\u22efF and phenyl-C\u2014H\u22efO inter\u00adactions , the latter leading to the dimeric aggregate.The title phosphanegold(I) thiol\u00adate, C The deviation of the P1\u2014Au\u2014S1 angle of 176.10\u2005(3)\u00b0 from the ideal 180\u00b0 is related to the close approach of the O1 atom, i.e. Au\u22efO1 = 2.986\u2005(2)\u2005\u00c5, as the O1 atom is directed towards the gold atom. The elongation of the C1\u2014S1 bond to 1.762\u2005(3)\u2005\u00c5 and the shortening of the C1\u2014N1 bond to 1.262\u2005(4)\u2005\u00c5 with respect to the comparable bonds in the neutral thio\u00adcarbamide mol\u00adecules, i.e. S=C(OMe)N(H)Ar R3PAu[SC(OR\u2032)=NAr]. However, a less common form is known whereby the N-bound aryl ring is orientated towards the gold atom rather than the alk\u00adoxy-oxygen atom =NPh], i.e. with Au\u22efO via pairwise fluoro\u00adbenzene-C\u2014H\u22efO1 and phenyl-C\u2014H\u22efF1 contacts Fig.\u00a02a). The dimeric aggregates are connected into a three-dimensional architecture by phenyl-C\u2014H\u22efS1 inter\u00adactions, with the phenyl-H atom involved in the latter inter\u00adaction, i.e. H13, also participating in a C\u2014H\u22ef\u03c0(fluoro\u00adbenzene) inter\u00adaction and so may be considered bifurcated. The two remaining contacts are of the type phenyl-C\u2014H\u22ef\u03c0 so the fluoro\u00adbenzene ring accepts two contacts, one to either side of the ring. A view of the unit-cell contents is shown in Fig.\u00a02b).Several directional inter\u00admolecular points of contact between mol\u00adecules are noted in the extended structure of (I)dnorm , where the cooperative phenyl-C13\u2014H13\u22ef\u03c0(C2\u2013C7) inter\u00adaction is shown as a distinctive orange \u2018pothole\u2019 on the shape-index-mapped Hirshfeld surface in Fig.\u00a04b). Although the phenyl-C22\u2014H22\u22ef\u03c0(C2\u2013C7) inter\u00adaction was not manifested on the Hirshfeld surface mapped over dnorm, this inter\u00adaction shows up as blue \u2018bump\u2019 and orange \u2018pothole\u2019 near the H22 atom and Cg1(C2\u2013C7) centroid, respectively, in Fig.\u00a05a). Simultaneously, a fluoro\u00adbenzene-C7\u2014H7\u22efC1(imine) contact, Table\u00a02dnorm surface in Fig.\u00a05b). The presence of the phenyl-C24\u2014H24\u22ef\u03c0(C11\u2013C16) contact is evidenced through faint red spots in Fig.\u00a06a) and the orange \u2018pothole\u2019 in Fig.\u00a06b) on the dnorm and shape-index mapped Hirshfeld surface, respectively. In addition to the C\u2014H\u22ef\u03c0 contacts listed in Table\u00a01In order to understand further the inter\u00adactions operating in the mol\u00adecular packing of (I)a), and those delineated into H\u22efH, H\u22efC/C\u22efH, H\u22efF/F\u22efH, H\u22efS/S\u22efH and H\u22efO/O\u22efH contacts are shown in Fig.\u00a08b)\u2013(f), respectively. The percentage contributions for the different inter\u00adatomic contacts to the Hirshfeld surface are summarized in Table\u00a03b) features a beak-shaped peak tipped at de + di \u223c2.3\u2005\u00c5. This tip corresponds to a methyl-H8C\u22efH33(phen\u00adyl) contact and has a distance 0.1\u2005\u00c5 shorter than the sum of their van de Waals radii, Table\u00a02c) with two symmetric spikes at de + di \u223c2.4\u2005\u00c5. The tips of pseudo-mirrored sharp spikes at de + di \u223c2.4\u2005\u00c5 represent the shortest H\u22efF/F\u22efH contacts (8.1%), Fig.\u00a08d), and correspond to the phenyl-C36\u2014H36\u22efF1 contact in Table\u00a01de + di \u223c2.7 and \u223c2.5\u2005\u00c5, respectively, Fig.\u00a08e) and (f), these types of contacts only contribute 6.9 and 3.2%, respectively, to the total inter\u00adatomic contacts. The accumulated contribution of the remaining six different inter\u00adatomic contacts is around 6.0% and these do not have a significant influence on the mol\u00adecular packing.The overall two-dimensional fingerprint plot of (I)et al., 2017via the long-range corrected \u03c9B97XD functional combining the D2 version of Grimme\u2019s dispersion model are listed in Table\u00a04The inter\u00adaction energies in the crystal of (I)\u22121). This is followed by the phenyl-C36\u2014H36\u22efF1 and phenyl-C3\u2014H3\u22efO1 inter\u00adactions , which lead to the dimeric aggregate in Fig.\u00a02a). The other directional contacts outlined in Supra\u00admolecular features contribute minor stabilization energies to the mol\u00adecular packing while the pairwise weak phenyl-C15\u2014H15\u22ef\u03c0(C21\u2013C26) and phenyl-C32\u2014H32\u22ef\u03c0(C11\u2013C16) inter\u00adactions, which were identified through the Hirshfeld surface analysis, have a greater stabilization energy .The greatest stabilization energy arises from the phenyl-C22\u2014H22\u22ef\u03c0(C2\u2013C7) and fluoro\u00adbenzene-C7\u2014H7\u22efC1(imine) inter\u00adactions =NC6H4Y-3]. Selected geometric parameters for these are given in Table\u00a056 residues, which vary by up to nearly 15\u00b0. Finally, there is an isostructural relationship between (I)Y = Cl compound 1H and 13C{1H} NMR spectra were recorded in CDCl3 solution on a Bruker Ascend 400\u2005MHz NMR spectrometer with chemical shifts relative to tetra\u00admethyl\u00adsilane; the 31P{1H} NMR spectrum was recorded in CDCl3 solution on the same instrument but with the chemical shift recorded relative to 85% aqueous H3PO4 as the external reference. IR spectra were measured on a Bruker Vertex 70v FTIR spectrophotometer from 4000 to 400\u2005cm\u22121. Elemental analyses were performed on a Leco TruSpec MicroCHN Elemental Analyser .All chemicals and solvents were used as sourced without further purification. Melting points were determined on a Biobase automatic melting point apparatus MP450 . M HCl. The resulting mixture was extracted using chloro\u00adform, yielding colourless crystals after 3 weeks standing. Yield: 0.421\u2005g (91%), m.p. 334.0\u2013334.5\u2005K. Analysis calculated for C8H8FNOS: C, 51.88; H, 4.35; N, 7.56%. Found: C, 51.49; H, 4.46; N, 7.42%. IR (cm\u22121): 3241 (br) \u03bd(N\u2014H), 1438 (s) \u03bd(C\u2014N), 1150 (s) \u03bd(C\u2014O), 1048 (s) \u03bd (C=S). 1H NMR : \u03b4 8.88 , 7.29\u20136.87 , 4.15 ppm. 13C{1H} NMR : \u03b4 189.4 (Cq), 162.8 , 138.5 (aryl-C1), 130.2 , 116.8 (aryl-C6), 112.2 , 109.1 (aryl-C2), 58.9 (OCH3) ppm.The thiol precursor, LH, was prepared from the reaction of 3-fluoro\u00adphenyl iso\u00adthio\u00adcyanate and MeOH in the presence of NaOH followed by the addition of excess 1 3PAuCl precursor was prepared from the reduction of KAuCl4 using sodium sulfite, followed by the addition of a stoichiometric amount of tri\u00adphenyl\u00adphosphane. The precipitate was used as isolated.The Ph3PAuCl in aceto\u00adnitrile (20\u2005ml), LH in aceto\u00adnitrile (20\u2005ml) was added and the solution was stirred for 3\u2005h. The solution was left for slow evaporation at room temperature, yielding colourless crystals after 2 weeks. Yield: 0.273\u2005g (85%), m.p. 408.0\u2013408.5\u2005K. Analysis calculated for C26H22AuFNOPS: C, 48.53; H, 3.45; N, 2.18%. Found: C, 48.73; H, 3.56; N, 1.97%. IR (cm\u22121): 1575 (s) \u03bd(C=N), 1122 (s) \u03bd(C\u2014O), 1100 (s) \u03bd(C\u2014S). 1H NMR : \u03b4 7.55\u20137.43 , 6.95\u20136.89 , 6.63\u20136.61 , 6.39\u20136.36 , 3.90 ppm. 13C{1H} NMR : \u03b4 165.4 (Cq), 163.2 , 152.8 , 134.3 , 131.7 , 129.7 , 129.3 , 129.1 , 117.8 , 109.3 , 109.1 , 55.5 (OCH3) ppm. 31P{1H} NMR : \u03b4 38.8 ppm.NaOH in water (5\u2005ml) was added to a suspension of PhUiso(H) set to 1.2\u20131.5Ueq(C). The maximum and minimum electron density peaks of 1.17 and 1.22\u2005e\u2005\u00c5\u22123, respectively, are located 0.97 and 0.69\u2005\u00c5, respectively, from the Au atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989020009469/hb7932sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020009469/hb7932Isup2.hklStructure factors: contains datablock(s) I. DOI: 2015568CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "PThe crystal structure of 4-allyl-2-meth\u00adoxy-6-nitro\u00adphenol, which crystallizes in the centrosymmetric space group 10H11NO4, which was synthesized via nitration reaction of eugenol with a mixture of nitric acid and sulfuric acid, consists of three independent mol\u00adecules of similar geometry. Each mol\u00adecule displays an intra\u00admolecular hydrogen bond involving the hydroxide and the nitro group forming an S(6) motif. The crystal cohesion is ensured by inter\u00admolecular C\u2014H\u22efO hydrogen bonds in addition to \u03c0\u2013\u03c0 stacking inter\u00adactions between the aromatic rings [centroid\u2013centroid distances = 3.6583\u2005(17)\u20134.0624\u2005(16)\u2005\u00c5]. The Hirshfeld surface analysis and the two-dimensional fingerprint plots show that H\u22efH (39.6%), O\u22efH/H\u22efO (37.7%), C\u22efH/H\u22efC (12.5%) and C\u22efC (4%) are the most important contributors towards the crystal packing.The asymmetric unit of the title compound, C The benzene rings of mol\u00adecules Mol-N2 and Mol-N3 are approximately parallel to each other [dihedral angle 10.60\u2005(7)\u00b0], and roughly perpendicular to that of Mol-N1 . A strong intra\u00admolecular O\u2014H\u22efO hydrogen bond involving a nitro O atom and the H atom of the hydroxide group forming an S(6) motif is observed in each mol\u00adecule \u2005\u00c5; Cg2\u22efCg3ii = 3.6613\u2005(18)\u2005\u00c5; Cg3\u22efCg3iii = 4.0624\u2005(16)\u2005\u00c5; symmetry codes: (i) 2\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z; (ii) 1\u00a0+\u00a0x, y, z; (iii) \u2212x, \u2212y, 1\u00a0\u2212\u00a0z].In the crystal, the mol\u00adecules are connected by inter\u00admolecular C12s Table\u00a01 and 3 \u25b8.et al., 2007Crystal Explorer 17.5 . The shape-index of the Hirshfeld surface is a tool to visualize the \u03c0\u2013\u03c0 stacking inter\u00adactions . The red spots in Fig.\u00a04a correspond to the strong C\u2014H\u22efO hydrogen-bond inter\u00adactions in the crystal structure; in Mol-N1 two of them involve the O atoms of the meth\u00adoxy (O1) and nitro (O3) groups as acceptors with allyl H atoms (C22B\u2013 H22B\u22efO1 and C12B\u2014H12B\u22efO3), while the other is due to the inter\u00adatomic inter\u00adaction between the aromatic H9 donor atom and the nitro O7 oxygen atom (C9\u2014H9\u22efO7). The longer O\u2014H\u22efO hydrogen bonds and O\u22efO inter\u00adactions are characterized by smaller red spots close to each other on the surface, where the faint red spot indicating the O\u2014H\u22efO inter\u00adactions is associated with the longest O\u22efO contact of 2.96\u2005(3)\u2005\u00c5 in Mol-N1 and Mol-N3. In Mol-N2, the red spots correspond to C\u2014H\u22efO (C9\u2014H9\u22efO7 and C12A\u2014H12A\u22efO12) and C\u2014H\u22efC (C20\u2014H20B\u22efC11A) hydrogen-bond inter\u00adactions. The corresponding fingerprint plots for each of the independent mol\u00adecules and for the entire asymmetric unit, showing characteristic pseudo-symmetric wings in the de and di diagonal axes, and those delineated into H\u22efH, O\u22efH/H\u22efO, C\u22efH/H\u22efC and C\u22efC contacts are illustrated in Fig.\u00a06b), which is reflected in the widely scattered points of high density due to the large hydrogen-atom content of the mol\u00adecule. The contribution from the O\u22efH/H\u22efO contacts (31.7%), corresponding to C\u2014H\u22efO and O\u2014H\u22efO inter\u00adactions, is represented by a pair of sharp spikes characteristic of a strong hydrogen-bond inter\u00adaction with de + di \u2243 2.5\u00c5 . In the absence of weak C\u2014H\u22ef\u03c0 inter\u00adactions in the crystal, the pair of characteristic wings in the fingerprint plot delineated into H\u22efC/C\u22efH contacts (7.7% contribution) have a symmetrical distribution of points , with the tips at de + di \u2243 2.65\u2005\u00c5. The distribution of points in the de = di \u2243 1.6\u2005\u00c5 range in the fingerprint plot delineated into C\u22efC contacts indicates the existence of weak \u03c0\u2013\u03c0 stacking inter\u00adactions between the phenyl rings, which are indicated by adjacent red and blue triangles in the shape-index map . The small contribution of the other weak inter\u00admolecular O\u22efO, N\u22efH/H\u22efN, C\u22efO/O\u22efC, C\u22efN/N\u22efC and N\u22efO/O\u22efN contacts has a negligible effect on the packing.In order to explore the nature of the inter\u00admolecular contacts and their role in the crystal packing, Hirshfeld surfaces as eluent. The reaction mixture was diluted with di\u00adchloro\u00admethane, washed with brine (3 \u00d7 10\u2005mL), dried over anhydrous Na2SO4 and concentrated under vacuum. The crude product was subjected to chromatography on a silica-gel column with n-hexa\u00adne/AcOEt (9:1 v/v) as eluent to afford the title compound as a reddish-orange liquid. Reddish-orange crystals formed spontaneously with a yield of 56%. Good quality crystals suitable for single crystal X-ray diffraction analysis were obtained by slow evaporation of an n-hexa\u00adne:AcOEt solution, m.p. = 317\u2013319\u2005K.In a 250\u2005mL flat-bottom flask containing a stirred solution of eugenol and di\u00adchloro\u00admethane (60\u2005mL), a mixture of concentrated sulfuric acid (0.78\u2005mL) and concentrated nitric acid (0.80\u2005mL) was added dropwise for 30\u2005min at 273\u2005K. The complete disappearance of the starting product was confirmed by means of thin layer chromatography using \u22121): 3235, 3080, 3016, 2971, 2910, 1638, 1537, 1392, 1331, 1262, 1128, 1059, 909, 763. The FT\u2013IR spectrum \u03b4 10.7 , 7.53 , 6.99 , 6.01\u20135.88 , 5.19\u20135.13 , 3.96 , 3.39\u20133.37 . 13C NMR \u03b4 149.87, 144.90, 135.95, 133.66, 131.24, 118.63, 117.16, 115.11, 114.29, 56.72, 39.41. FTMS\u2013ESI, m/z: 208.04616 (100%) [C10H11NO4].Uiso(H) = 1.2 Ueq(C) or 1.5Ueq(C) for methyl H atoms. A rotating model was used for the methyl groups. The hydroxyl H atoms were located in a difference-Fourier map and refined freely. The two allyl groups of Mol-N2 and Mol-N3 are disordered over two sets of sites with refined occupancy ratios of 0.648\u2005(8):0.352\u2005(8) and 0.668\u2005(9):0.332\u2005(9) respectively. One outlier (100) was omitted in the cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020002601/rz5270sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020002601/rz5270Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020002601/rz5270Isup3.cmlSupporting information file. DOI: 1986157CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Its crystal structure was determined at 100\u2005K. Both weak intra\u00admolecular O\u2014H\u22efO and inter\u00admolecular O\u2014H\u22efS hydrogen bonding can be observed.The title compound is an example of an 22H22O2S2, 1, represents an example of an ortho-vanillin-based functionalized di\u00adthio\u00adether, which could be useful as a potential chelating ligand or bridging ligand for coordination chemistry. This di\u00adthio\u00adacetal 1 crystallizes in the ortho\u00adrhom\u00adbic space group Pbca. The phenyl rings of the benzyl groups and that of the vanillin unit form dihedral angles of 35.38\u2005(6) and 79.77\u2005(6)\u00b0, respectively. The crystal structure, recorded at 100\u2005K, displays both weak intra\u00admolecular O\u2014H\u22efO and inter\u00admolecular O\u2014H\u22efS hydrogen bonding.The title compound, C R)(H)\u2013S\u2013 motif are synthesized by nucleophilic substitution of geminal dihalides X\u2013C(R)(H)\u2013X in the presence of thiol\u00adate RS\u2212 nSH , yielding geminal di\u00adthio ethers, also called acyclic and cyclic thio\u00adacetals Fe(CO)2(\u03ba1-BzSCH2SBz)]+, the 1:1 adduct [Hg2(NO3)2\u00b7BzSCH2SBz)], the dinuclear PdI complex [ClPd(\u03bc2-BzSCH2SBz)2PdCl], and the monodimensional coordination polymer [Ag2(BzSCH2SBz)2](ClO4)2 built upon dinuclear [Ag(\u03bc2-BzSCH2SBz)2Ag]2+ units as the starting material. This hy\u00addroxy\u00adlated aldehyde is present in the extracts and essential oils of many plants. Several papers describe also its use (in its deprotonated vanillinato form or as a Schiff base-derived ligand) in coordination chemistry and 1.8189\u2005(12)\u2005\u00c5 are comparable with those of [BzSC(H)(C6H4NO2-p)SBz] [1.823\u2005(3) and 1.8262\u2005(19)\u2005\u00c5], but are elongated compared with those of bis\u00ad(benzyl\u00adsulfan\u00adyl)methane (CSD TUQPAX) , but considerably more acute than in [BzSCH2SBz] [117.33\u2005(7)\u00b0]. There is a weak intra\u00admolecular O1\u22efH2 contact of 2.17\u2005(2)\u2005\u00c5 between the H atom of the phenolic hydroxyl group and the O-atom of the meth\u00adoxy group SBz] , O2\u22efS1 = 3.1315\u2005(13)\u2005\u00c5] similar to those reported for 4--1,2-benzene\u00addiol , while the O\u2014H\u22efS angle is more acute [139.0\u2005(17) \u00b0] Fig.\u00a03. This O2The benzylic methyl\u00adene group on sulfur atom S2 inter\u00adacts with the \u03c0-cloud of the phenyl part of the vanillin unit through a C\u2014H\u22ef\u03c0 inter\u00adaction Table\u00a01. The secet al., 2016et al., 2003et al., 2005et al., 2013et al., 2013et al., 2018et al., 20131.There are several other examples of structurally characterized related di\u00adthioethers bearing hy\u00addroxy substituents that give rise to the formation of supra\u00admolecular networks. Selected examples found in the Cambridge Structural Database -1,3-di\u00adthiane (WADROY) and 2-(3-hy\u00addroxy\u00adphen\u00adyl)-1,3-di\u00adthiane (KALJUD) exhibit, like 1, only inter\u00admolecular O\u2014H\u22efS hydrogen bonding, the para-derivative 2-(4-hy\u00addroxy\u00adphen\u00adyl)-1,3-di\u00adthiane (KALKAK) features solely inter\u00admolecular phenolic O\u2014H\u22efH bonding .Note that in di\u00adthio\u00adether compounds with phenolic aryl groups as encountered in The reaction scheme for the synthesis of the title compound is illustrated in Fig.\u00a043 (10\u2005mL) and extracted with di\u00adchloro\u00admethane (3 \u00d7 10\u2005mL). The combined extracts were washed with H2O (3 \u00d7 20\u2005mL) and dried over Na2SO4. Evaporation of the solvent in vacuo gave a solid product, which was further purified by column chromatography. The product was obtained as a white solid, Yield: 83% (430\u2005mg). X-ray quality crystals were obtained by keeping a di\u00adchloro\u00admethane:hexane (1:1) mixture of 1 at 278\u2005K for 3\u20134\u2005d. 1H NMR \u03b4 7.26\u20137.19 , 6.86 , 6.78 , 5.83 , 5.13 , 3.88 , 3.79 , 3.64 .13C{1H} NMR \u03b4 146.5 (CqOH), 142.8 (CqOCH3), 137.8 (SCH2Cq), 129.1 (SCH2CCH), 128.4 (SCH2CCHCH), 126.9 (SCH2CCHCHCH), 125.3 (S2CHCq), 120.9 (S2CHCqCH), 119.9 (S2CHCqCHCH), 110.0 (CHCqOCH3), 56.1 (OCH3), 44.8 (S2CH), 36.7 (S2CH2). IR (ATR) cm\u22121: 3419 (O\u2014H), 1430-1612 (C=C). 1054 and 1264 (C\u2014O), 766 (C\u2014S). HRMS: (ESI) m/z calculated for C22H22O2S2Na [M + Na]+ 405.0953, found 405.0965.3-Meth\u00adoxy\u00adsalicyl\u00adaldehyde , benzyl mercaptan , and conc. HCl (2\u2005mL) were added to a flask at 273\u2005K. The mixture was stirred for 60\u2005min at room temperature. After the reaction was complete, the resulting mixture was neutralized with 10% aq NaHCOUiso(H) = 1.2Ueq(C) for CH2 and CH hydrogen atoms and Uiso(H) = 1.5Ueq(C-meth\u00adyl). The phenolic proton H2 was refined independently.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020002091/vm2228sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020002091/vm2228Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020002091/vm2228Isup3.cmlSupporting information file. DOI: 1983985CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular and crystal structure as well as the Hirshfeld surface were analysed for this benzo\u00adthia\u00adzine derivative with potential analgesic activity. 10H8ClNO5S, which has potential analgesic activity, crystallizes in space group P21/n. The benzo\u00adthia\u00adzine ring system adopts an inter\u00admediate form between sofa and twist-boat conformations. The coplanarity of the ester substituent to the bicyclic fragment is stabilized by an O\u2014H\u22efO intra\u00admolecular hydrogen bond. In the crystal, hydrogen bonds of type N\u2014H\u22efO(SO2) link the mol\u00adecules into zigzag chains extending along the b-axis direction. Neighbouring chains are linked by both O\u2014H\u22efCl and C\u2014H\u22efCl inter\u00adactions. A Hirshfeld surface analysis was used to compare different types of inter\u00admolecular inter\u00adactions, giving contributions of O\u22efH/H\u22efO = 42.0%, C\u22efH/H\u22efC = 17.3%, Cl\u22efH/H\u22efCl = 14.2%, H\u22efH = 11.1%.The title compound, C H-2\u03bb6,1-benzo\u00adthia\u00adzine-3-carbox\u00adyl\u00adates are known to be highly active analgesics \u2005\u00c5, \u0398 = 111.6\u2005(5)\u00b0, \u03a8 = 192.1\u2005(6)\u00b0. The ester substit\u00aduent is essentially coplanar to the C7\u2014C8 endocyclic double bond [the C7\u2014C8\u2014C9\u2014O2 torsion angle is 3.0\u2005(7)\u00b0] as a result of the stabilizing influence of the O1\u2014H1O\u22efO2 intra\u00admolecular hydrogen bond (Table\u00a01S(6) in terms of graph-set theory since the six atoms comprise a intra\u00admolecular hydrogen-bonded motif. The formation of the O\u2014H\u22efO hydrogen bond causes some elongation of the C9\u2014O2 and C7\u2014C8 bonds as compared with typical values of 1.210\u2005\u00c5 (Csp2=O bond) and 1.326\u2005\u00c5 (Csp2\u2014Csp2 bond), respectively for the same reason. The methyl group is located in an anti-periplanar position to the C8\u2014C9 bond [the C10\u2014O3\u2014C9\u2014C8 torsion angle is \u2212178.4\u2005(4)\u00b0]. The noticeable steric repulsion between chlorine and the hydroxyl group . The crs Table\u00a01 in the was added, the mixture was boiled and stored for 15\u2005h at room temperature. The reaction mixture was diluted with cold water and acidified with 1\u2005N HCl to pH = 4. The solid methyl 5-chloro-4-hy\u00addroxy-2,2-dioxo-1H-2\u03bb6,1-benzo\u00adthia\u00adzine-3-carboxyl\u00adate was filtered, washed with water, and dried in air. Yield 2.43g (84%); colourless crystals; m.p. 464\u2013466\u2005K.Methyl (chloro\u00adsulfon\u00adyl)acetate was added dropwise under stirring to a solution of methyl 6-chloro\u00adanthranilate and tri\u00adethyl\u00adamine in CHK) Fig.\u00a05. After 1Uiso(H) = 1.5Ueq for methyl and hydroxyl groups and with C\u2014H = 0.93\u2005\u00c5, N\u2014H = 0.88\u2005\u00c5, Uiso(H) = 1.2Ueq for all other hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020012566/pk2647sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020012566/pk2647Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020012566/pk2647Isup3.cmlSupporting information file. DOI: 2032151CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Bnz\u22efOThz (Bnz = benzene and Thz = thia\u00adzine) hydrogen bonds form chains of mol\u00adecules extending along the a-axis direction which are connected to their inversion-related counterparts by C\u2014HBnz\u22efClDchlphy hydrogen bonds and C\u2014HDchlphy\u22ef\u03c0 (ring) inter\u00adactions. These double chains are further linked by C\u2014HDchlphy\u22efOThz hydrogen bonds to form stepped layers approximately parallel to (012).The title compound contains 1,4-benzo\u00adthia\u00adzine and 2,4-di\u00adchloro\u00adphenyl\u00admethyl\u00adidene units in which the di\u00adhydro\u00adthia\u00adzine ring adopts a screw-boat conformation. In the crystal, inter\u00admolecular C\u2014H 24H27Cl2NOS, contains 1,4-benzo\u00adthia\u00adzine and 2,4-di\u00adchloro\u00adphenyl\u00admethyl\u00adidene units in which the di\u00adhydro\u00adthia\u00adzine ring adopts a screw-boat conformation. In the crystal, inter\u00admolecular C\u2014HBnz\u22efOThz (Bnz = benzene and Thz = thia\u00adzine) hydrogen bonds form chains of mol\u00adecules extending along the a-axis direction, which are connected to their inversion-related counterparts by C\u2014HBnz\u22efClDchlphy hydrogen bonds and C\u2014HDchlphy\u22ef\u03c0 (ring) inter\u00adactions. These double chains are further linked by C\u2014HDchlphy\u22efOThz hydrogen bonds, forming stepped layers approximately parallel to (012). The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (44.7%), C\u22efH/H\u22efC (23.7%), Cl\u22efH/H\u22efCl (18.9%), O\u22efH/H\u22efO (5.0%) and S\u22efH/H\u22efS (4.8%) inter\u00adactions. Hydrogen-bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Computational chemistry indicates that in the crystal, C\u2014HDchlphy\u22efOThz, C\u2014HBnz\u22efOThz and C\u2014HBnz\u22efClDchlphy hydrogen-bond energies are 134.3, 71.2 and 34.4\u2005kJ\u2005mol\u22121, respectively. Density functional theory (DFT) optimized structures at the B3LYP/6\u2013311\u2005G level are compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap. The two carbon atoms at the end of the nonyl chain are disordered in a 0.562\u2005(4)/0.438\u2005(4) ratio.The title compound, C These mol\u00adecules exhibit a wide range of biological applications, indicating that the 1,4-benzo\u00adthia\u00adzine moiety is a potentially useful template in medicinal chemistry research with therapeutic applications in the anti\u00admicrobial (Armenise B (S1/N1/C1/C6\u2013C8), adopts a screw-boat conformation with puckering parameters QT = 0.5581\u2005(16)\u2005\u00c5, \u03b8 = 69.76\u2005(18)\u00b0 and \u03c6 = 334.3\u2005(2)\u00b0. The planar rings, A (C1\u2013C6) and C (C10\u2013C15) are oriented at a dihedral angle of 88.45\u2005(7)\u00b0. Atoms Cl1, Cl2 and C9 are almost co-planar with ring C being displaced by 0.0247\u2005(6), \u22120.0732\u2005(9) and \u22120.0274\u2005(2)\u2005\u00c5, respectively.The title compound contains 1,4-benzo\u00adthia\u00adzine and 2,4-di\u00adchloro\u00adphenyl\u00admethyl\u00adidene units Fig.\u00a01, in whicBnz\u22efOThz (Bnz = benzene and Thz = thia\u00adzine) hydrogen bonds link the mol\u00adecules, forming chains extending along the a-axis direction, which are connected to their inversion-related counterparts by C\u2014HBnz\u22efClDchlphy hydrogen bonds and C\u2014HDchlphy\u22ef\u03c0 (ring) inter\u00adactions Table\u00a01 and 3 \u25b8.Crystal Explorer 17.5 . The two pairs of wings in the fingerprint plot delineated into Cl\u22efHl/H\u22efCl contacts have an unsymmetrical distribution of points due to a third wing, with the edges at de + di = 2.74\u2005\u00c5 (for the long wing), de + di = 2.92\u2005\u00c5 (for the short wing) and de + di = 3.53\u2005\u00c5 . The pair of wings in the fingerprint plot delineated into O\u22efH/H\u22efO contacts has a pair of spikes with the tips at de + di = 2.22\u2005\u00c5. Finally, the wings in the fingerprint plot delineated into S\u22efH/H\u22efS contacts have the tips at de + di = 2.99\u2005\u00c5.In order to visualize the inter\u00admolecular inter\u00adactions in the crystal of the title compound, a Hirshfeld surface (HS) analysis is the sum of electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies were calculated to be \u221253.7 (Eele), \u221213.6 (Epol), \u2212161.9 (Edis), 119.0 (Erep) and \u2212134.3 (Etot) for C\u2014HDchlphy\u22efOThz, 25.6 (Eele), \u22125.7 (Epol), \u221262.1 (Edis), 23.1 (Erep) and \u221271.2 (Etot) -4-nonyl-3,4-di\u00adhydro-2H-1,4-benzo\u00adthia\u00adzin-3-one ring.The optimized structure of the title compound, (I)et al., 2016II gave 14 hits.A search in the Cambridge Structural Database , 29\u00b0 (APAJUY), 28\u00b0 (SAVTUH), 26\u00b0 (WOCFUS) and 25\u00b0 (COGRUN). By contrast, in both EVIYIT and OMEGEU, the benzo\u00adthia\u00adzine unit is nearly planar with the corresponding dihedral angle being about 4\u00b0.The largest set contains Z)-2--2H-1,4-benzo\u00adthia\u00adzin-3(4H)-one (1.5\u2005mmol), potassium carbonate (2.7\u2005mmol) and tetra-n-butyl ammonium bromide (0.14\u2005mmol) in DMF (20\u2005mL) was added 1-bromo\u00adnonane (2.6\u2005mmol). Stirring was continued at room temperature for 24\u2005h. The mixture was filtered and the solvent removed. The residue obtained was washed with water. The organic compound was chromatographed on a column of silica gel with ethyl acetate\u2013hexane (9/1) as eluent. Colourless crystals were isolated when the solvent was allowed to evaporate (yield = 79%).To a solution of (Uiso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989020001036/lh5943sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020001036/lh5943Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020001036/lh5943Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020001036/lh5943Isup4.cmlSupporting information file. DOI: 1980073CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dihedral angle between the mean planes of the benzene ring and that of the twisted piperidine ring is 39.89\u2005(7)\u00b0. In the crystal, weak C\u2014H\u22efO inter\u00adactions link the mol\u00adecules into infinite chains along the 13H16ClNO, contains a methyl\u00adpiperidine ring in the stable chair conformation. The mean plane of the twisted piperidine ring subtends a dihedral angle of 39.89\u2005(7)\u00b0 with that of the benzene ring. In the crystal, weak C\u2014H\u22efO inter\u00adactions link the mol\u00adecules along the a-axis direction to form infinite mol\u00adecular chains. H\u22efH inter\u00adatomic inter\u00adactions, C\u2014H\u22efO inter\u00admolecular inter\u00adactions and weak dispersive forces stabilize mol\u00adecular packing and form a supra\u00admolecular network, as established by Hirshfeld surface analysis.The title compound, C The C1\u2014N1\u2014C6\u2014O1 and O1\u2014C6\u2014C7\u2014C8 torsion angles are \u2212167.4\u2005(2) and 50.7\u2005(3)\u00b0, respectively. The C1\u2014C2\u2014C3\u2014C13 torsion angle [177.7\u2005(2)\u00b0] reveals the anti-periplanar (+ap) orientation of the methyl group with respect to the piperidine ring.The mol\u00adecular structure of the title compound, which features a chloro\u00adbenzene ring and a methyl\u00adpiperidine ring, is shown in Fig.\u00a01x\u00a0\u2212\u00a01, y, z), forming chains parallel to the a axis (Table\u00a01a). Weak C\u2014H\u22ef\u03c0 close contacts between H5A and the benzene ring of an adjacent mol\u00adecule provide linkage between inversion-related chains (Table\u00a01b). Analysis of the Hirshfeld surface = 1.5Ueq(C-meth\u00adyl) or 1.2Ueq(C) for all other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020001930/jj2221sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020001930/jj2221Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020001930/jj2221Isup3.cmlSupporting information file. DOI: 1973841CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The packing of the title mol\u00adecular salt, in which the tin atom lies on a crystallographic inversion centre, is dominated by N\u2014H\u22efCl hydrogen bonds. 5H7N2)2[SnCl6], the cation is protonated at the pyridine N atom and the complete dianion is generated by a crystallographic centre of symmetry. In the crystal, N\u2014H\u22efCl hydrogen bonds link the components into a three-dimensional network built up from the stacking of alternate cationic and anionic layers. The nature of the inter\u00admolecular inter\u00adactions has been analysed in terms of the Hirshfeld surfaces of the cations and the anions. The thermal behaviour and the Raman spectrum of the title compound are reported.In the title mol\u00adecular salt, (C For example, in these hybrid materials, the organic part can have non-linear optical properties and an organic cation +2\u00b7[SnCl6]2\u2212, crystallizes in the triclinic space group PCompound (I)P Fig.\u00a01.et al., 2018i.e. none of the chloride ions are bridging although they do accept N\u2014H\u22efCl hydrogen bonds from the organic cations, which ensures charge balance.In the synthesis, the oxidation number of tin changes from +2 to +4 such that the resultant tin(IV) atom is hexa\u00adcoordinated by chlorine atoms, generating a weakly distorted octa\u00adhedron in which the metal ion lies on a crystallographic inversion centre: the length of the Sn\u2014Cl bonds varies from 2.4216\u2005(4) to 2.4474\u2005(5)\u2005\u00c5. As for the Cl\u2014Sn\u2014Cl angles, the discrepancy of about \u00b1 1\u00b0 [89.109\u2005(18)\u201390.805\u2005(16)\u00b0] compared to the 90\u00b0 value angle of a regular octa\u00adhedron shows that the angular distortion is very small. These values are comparable to those of the same anion associated with other types of cations to 1.405\u2005(3)\u2005\u00c5 and the C\u2014N bond lengths are 1.341\u2005(3)\u2005\u00c5 and 1.344\u2005(2)\u2005\u00c5. The values of the C\u2014C\u2014C angles in the pyridinium ring vary from 118.9\u2005(2) to 120.9\u2005(2)\u00b0 whereas the C\u2014N\u2014C angle is 124.30\u2005(18)\u00b0: the larger angle can be attributed to the protonation of the N atom. These values are comparable with those of the same cation associated with other types of anions (Rao a). The inter\u00admolecular inter\u00adactions in (I)PLATON in the crystal of (I)A\u22efCl1 and N2\u2014H2B\u22efCl2 hydrogen bonds generates a chain of rings propagating along the [001] direction with a graph-set pattern of et al., 1990b). The cohesion between chains is ensured by \u03c0-stacking inter\u00adactions between centrosymmetrically related aromatic rings of the cations: Cg1\u22efCg1 = 3.552\u2005(13)\u2005\u00c5; inter-planar angle \u03b1 = 0.0\u2005(11)\u00b0; slippage = 1.246\u2005\u00c5. We also note the presence of a Y\u2014X\u22efCg1 type inter\u00adaction between Sn1\u2014Cl2 and Cg1 at an X\u22efCg distance of 3.6581\u2005(11)\u2005\u00c5 [Fig.\u00a02a)].The combination of N2\u2014H2Crystal Explorer 17 a)] shows red spots corresponding to H\u22efCl/Cl\u22efH close contacts, which are due to the N\u2014H\u22efCl hydrogen bonds. The presence of the adjacent red and blue triangles in Fig.\u00a03b) demonstrates the presence of the Cg1\u22efCg1 and Sn\u2014Cl2\u22efCg1 inter\u00adactions. The contribution of different kinds of inter\u00adatomic contacts to the Hirshfeld surfaces of the individual cations and anions is shown in the fingerprint plots in Fig.\u00a04Y\u2014X\u22ef type (N\u22efCl and C\u22efCl), 6.6% of \u03c0\u2013\u03c0 stacking type (C\u22efC and C\u22efN/N\u22efC), 15.6% of C\u2014H\u22ef\u03c0 type (C\u22efH/H\u22efC), 6.2% of N\u2014H\u22ef\u03c0 type (N\u22efH/H\u22efN) and 21.1% of H\u22efH van der Waals inter\u00adactions. The two-dimensional fingerprint analysis for the anionic moieties reveals that hydrogen bonds (Cl\u22efH) represent 93.8%, Y\u2014X\u22ef\u03c0 type inter\u00adactions represent 4.4% (Cl\u22efN and Cl\u22efC) and van der Waals inter\u00adactions of the Cl\u22efCl type represent 1.8% of the surface contacts.The Hirshfeld surface 2\u2212 anions are associated with special positions and an organic\u2013inorganic layered structure lying parallel to the (001) plane results.A search of the Cambridge Structural Database .Crystalline cohesion in RIGDER and (I)2 atmosphere at a heating rate of 10\u00b0C min\u22121 in the temperature range from 25 to 500\u00b0C. The thermogram of (I)supporting information) shows that the compound loses 64.4% of its mass in the temperature range of 270\u2013304\u00b0C. The mass loss can be attributed to the degradation of the organic entity and two chlorine atoms 1H NMR (\u03b4 ppm), 400\u2005MHz, CDCl3): 8.16 , 7.95\u20137.89 , 7.03 , 6.84 . 13C NMR (\u03b4 ppm), 125\u2005MHz, CDCl3): 154.6 (quat C Py), 144.4 (CH Py), 136.1 (CH Py), 113.8 (CH Py), 112.5 (CH Py).Tin(II) chloride dihydrate (2.25\u2005mmol) was mixed with 2-amino\u00adpyridine (0.94\u2005mmol) and a few drops of hydro\u00adchloric acid in an aliquot of distilled water in 1:1 molar ratio was added. After stirring, the mixture was poured into a vial that was put in an oven for three days at 393\u2005K. Upon cooling, prism-shaped crystals of (I)\u22121. The Py, \u03bd, \u03b4, \u03b3 and \u03c4 are: pyridine ring, stretching, in-plane bending, out-of-plane bending and torsion, respectively. RS (cm\u22121): 3334 \u03bd(N\u2014H), 3215 \u03bd (N\u2014H), 3106 \u03bd(C\u2014H), 1657 \u03bd(py)+\u03b4(N\u2014H)+\u03b4(NH2), 1620 \u03bd(py), 1542 \u03bd(py), 1472 \u03bd(py)+\u03b4(C\u2014H), 1412 \u03bd(py)+\u03b4(C\u2014H), 1378 \u03bd(py)+\u03b4(C\u2014H), 1324 \u03bd(py)+\u03b4(C\u2014H), 1239 \u03bd(py)+\u03b4(C\u2014H), 1164 \u03b4(py)+\u03b4(C\u2014H), 1120 \u03b4(py)+\u03b4(C\u2014H), 996 \u03b4(py), 846 Pyridine ring breathing+\u03b3(C\u2014H), 623 \u03b3(py), 551 \u03b3(py), 406 \u03b3(py), 384 \u03b3(py), 305 \u03bd1 (Sn\u2014Cl), 216 \u03bd2 (Sn\u2014Cl), 106 \u03c4(py)+ \u03bd(N\u2014H\u22efCl) Uiso(H) = 1.2Ueq(C)]; the pyridine N\u2014H atom was located in a difference map and its position was freely refined.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698902000941X/hb7922sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902000941X/hb7922Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902000941X/hb7922sup3.jpgSupporting information file. DOI: Click here for additional data file.10.1107/S205698902000941X/hb7922sup4.docxSupporting information file. DOI: 1904730CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Theoretical calculations suggested that the new bis\u00ad(hy\u00addroxy\u00adimine) will exhibit histone de\u00adacetyl\u00adase SIRT2, histone de\u00adacetyl\u00adase class III and histone de\u00adacetyl\u00adase SIRT1 activities, and will act as inhibitor to aspulvinone di\u00admethyl\u00adallyl\u00adtransferase, de\u00adhydro- 47H32N2O2\u00b7C6H4Cl2, contains two anil fragments in the enol\u2013enol form, exhibiting intra\u00admolecular O\u2014H\u22efN hydrogen bonds. The two hy\u00addroxy\u00adnaphthalene ring systems are approximately parallel to each other with a dihedral angle of 4.67\u2005(8)\u00b0 between them, and each ring system makes a large dihedral angle [55.11\u2005(11) and 48.50\u2005(10)\u00b0] with the adjacent benzene ring. In the crystal, the bis\u00ad(anil) mol\u00adecules form an inversion dimer by a pair of weak C\u2014H\u22efO inter\u00adactions. The dimers arrange in a one-dimensional column along the b axis via another C\u2014H\u22efO inter\u00adaction and a \u03c0\u2013\u03c0 stacking inter\u00adaction between the hy\u00addroxy\u00adnaphthalene ring system with a centroid\u2013centroid distance of 3.6562\u2005(16)\u2005\u00c5. The solvent 1,2-di\u00adchloro\u00adbenzene mol\u00adecules are located between the dimers and bind neighbouring columns by weak C\u2014H\u22efCl inter\u00adactions. Theoretical prediction of potential biological activities was performed, which suggested that the title anil compound can exhibit histone de\u00adacetyl\u00adase SIRT2, histone de\u00adacetyl\u00adase class III and histone de\u00adacetyl\u00adase SIRT1 activities, and will act as inhibitor to aspulvinone di\u00admethyl\u00adallyl\u00adtransferase, de\u00adhydro-l-gulonate deca\u00adrboxylase and gluta\u00adthione thiol\u00adesterase.The bis\u00ad(anil) mol\u00adecule of the title compound, C According to the PASS program \u2013 computer prediction of biological activities and gluta\u00adthione thiol\u00adesterase (71%).Schiff bases formed by the condensation of salicyl\u00adaldehydes with amines are also known as anils. They often exhibit potent anti\u00adbacterial, anti\u00adproliferative and anti\u00adtoxic properties forms large dihedral angles of 78.80\u2005(10) and 61.41\u2005(9)\u00b0, respectively, with the benzene C14\u2013C19 and C31\u2013C36 rings. Each hy\u00addroxy\u00adnaphthalene ring system also forms a large dihedral angle with the adjacent benzene ring . Both fragments of the hy\u00addroxy\u00adnaphthalene Schiff bases are in the enol form, forming intra\u00admolecular O\u2014H\u22efN hydrogen bonds (Table\u00a01In the title bis\u00ad(anil) mol\u00adecule, two hy\u00addroxy\u00adnaphthalene ring systems are approximately parallel to each other with a dihedral angle of 4.67\u2005(8)\u00b0 between them Fig.\u00a01. The 9H-s Table\u00a01.via a pair of weak C\u2014H\u22efO inter\u00adactions . Di\u00adchloro\u00adbenzene mol\u00adecules are located between the dimers and bind the neighboring columns by weak C\u2014H\u22efCl inter\u00adactions mol\u00adecules form an inversion dimer s Table\u00a01.et al., 2016via the condensation of the corresponding di\u00adamine and an appropriate hy\u00addroxy\u00adaldehyde dianiline according to the literature = 1.2Ueq(C)], while the H atoms of the OH groups were localized in a difference-Fourier map and refined with Uiso(H) = 1.5Ueq(O).Crystal data, details of data collection, and results of structure refinement are summarized in Table\u00a0210.1107/S2056989020012104/is5554sup1.cifCrystal structure: contains datablock(s) general, I. DOI: 10.1107/S2056989020012104/is5554Isup2.hklStructure factors: contains datablock(s) I. DOI: 2019265CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Compounds 1 and 2 crystallize, respectively, in space group P21/c with Z\u2032 = 2 and in space group Pbca with Z\u2032 = 1. The crystals of compound 1 contain neutral and anionic Bmphfp mol\u00adecules, and form a one-dimensional hydrogen-bonded chain motif. The crystals of compound 2 contain anionic Bmphfp mol\u00adecules, which form a complex three-dimensional hydrogen-bonded network with the ethyl\u00adenedi\u00adamine and water mol\u00adecules.The crystal structures of two salt crystals of 2,2-bis\u00ad(4-methyl\u00adphen\u00adyl)hexa\u00adfluoro\u00adpropane (Bmphfp) with amines, namely, dipyridinium 4,4\u2032-dibenzoate 4,4\u2032-di\u00adbenzoic acid, 2C Inter\u00adestingly, one of the two Bmphfp mol\u00adecules is neutral and the other is anionic. The C\u2014O bond lengths of the carb\u00adoxy groups are summarized in Table\u00a01A and B are in the neutral and divalent anionic forms, respectively. The two benzene rings are twisted with respect to each other, forming dihedral angles of 72.19\u2005(6) and 69.98\u2005(6)\u00b0. On the other hand, compound 2 crystallizes in the ortho\u00adrhom\u00adbic space group Pbcn. The asymmetric unit comprises one Bmphfp anion, one ethyl\u00adenedi\u00adammonium cation and one water mol\u00adecule . The C\u2014O bond lengths shown in Table\u00a01Compound it Fig.\u00a01a. Interle Fig.\u00a01b. The C1, the pyridine mol\u00adecules form strong N\u2014H\u22efO hydrogen bonds with the carboxyl groups of Bmphfp mol\u00adecule B (Table\u00a02a). The neutral (A) and anionic (B) Bmphfp mol\u00adecules form a one-dimensional hydrogen-bonded chain motif along the a-axis direction . The lengths of the negative charge-assisted O\u2014H\u22efO hydrogen bonds, 2.5732\u2005(13) and 2.5125\u2005(13)\u2005\u00c5, are shorter than in the common carboxyl dimer [2.643\u2005\u00c5; the mean value calculated from 505 research hits in the Cambridge Structural Database \u2005\u00c5 (C\u22efF) and 2.696\u2005(1)\u2005\u00c5 (F\u22efF), respectively.In the crystal of compound B Table\u00a02a. The non Fig.\u00a02b. The l2, one carb\u00adoxy\u00adlic group of the Bmphfp mol\u00adecule is linked to an ethyl\u00adenedi\u00adammonium cation by two N\u2014H\u22efO hydrogen bonds. The N\u22efO inter\u00adatomic distances are 2.7749\u2005(14) and 2.8015\u2005(14)\u2005\u00c5, respectively (Table\u00a03a). Therefore, five of the six hydrogen-atom donors of the ethyl\u00adenedi\u00adammonium cations are connected to Bmphfp mol\u00adecules, resulting in a complex three-dimensional hydrogen-bonding network. The water mol\u00adecule is linked to both Bmphfp and ethyl\u00adenedi\u00adamine mol\u00adecules via two O\u2014H\u22efO and one N\u2014H\u22efO hydrogen bonds. Thus, the water mol\u00adecules are highly stabilized by these inter\u00admolecular inter\u00adactions in the crystal structure . Weak C\u2014H\u22efF and F\u22efF inter\u00adactions are observed between Bmphfp mol\u00adecules, resulting inter\u00adatomic distances of 3.493\u2005(1)\u2005\u00c5 (C\u22efF) and 2.890\u2005(1)\u2005\u00c5 (F\u22efF), respectively. In compound 2, the Bmphfp mol\u00adecules do not form a discrete 1-D hydrogen bond chain motif as observed in compound 1 because the one carboxyl group is terminated by an ethyl\u00adenedi\u00adamine mol\u00adecule.In the crystal of compound y Table\u00a03. The othy Table\u00a03a. Therere Fig.\u00a03b. Weak et al., 2007CrystalExplorer17 compared to mol\u00adecule A. Thus, mol\u00adecule B is more closely packed with the surrounding mol\u00adecules in the crystal than mol\u00adecule A. This may be due to the difference in the ionic state between neutral mol\u00adecule A and anionic mol\u00adecule B. Compound 1 (mol\u00adecule A and B) has strong hydrogen-bonding inter\u00adactions, with similar but slightly weaker inter\u00adactions for compound 2. The contributions to the Hirshfeld surface for 2 are listed in Table\u00a06Hirshfeld surfaces benzene hexa\u00adfluoro\u00adpropane, pyridine and ethyl\u00adenedi\u00adamine were purchased from TCI Co., Ltd. (Japan). 2,2-Bis(4-methyl\u00adphen\u00adyl)hexa\u00adfluoro\u00adpropane was dissolved in methanol 10\u2005mL. The Bmphfp solution was mixed into 5\u2005mL of a 1.0 sp2\u2014H = 0.86, Nsp3\u2014H = 0.89\u2005\u00c5 with Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S2056989020005575/lh5959sup1.cifCrystal structure: contains datablock(s) 1, 2, global. DOI: 10.1107/S2056989020005575/lh59591sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989020005575/lh59592sup3.hklStructure factors: contains datablock(s) 2. DOI: 1998170, 1998169CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The PtII atoms in both mol\u00adecules adopt distorted square-planar geometries, coordinated by one C and two N atoms from the tridentate 2\u2032,6\u2032-di\u00adfluoro-6-[3-(pyridin-2-yl\u00adoxy)phen\u00adyl]-2,3\u2032-bi\u00adpyridine ligand and a chloride anion: the C and Cl atoms are trans. In the crystal, C\u2014H\u22efCl/F hydrogen bonds, F\u22ef\u03c0 and weak \u03c0\u2013\u03c0 stacking inter\u00adactions between adjacent A and B mol\u00adecules and between pairs of inversion-related B mol\u00adecules lead to the formation of a two-dimensional supra\u00admolecular network lying parallel to the ab plane. The sheets are stacked along the c-axis direction and linked by F\u22ef\u03c0 and weak \u03c0\u2013\u03c0 stacking inter\u00adactions between pairs of inversion-related A mol\u00adecules, forming a three-dimensional supra\u00admolecular network. The photoluminescence quantum efficiency of the title compound in the blue\u2013green region of the visible region (\u03bbmax = 517 and 544\u2005nm) is estimated to be \u223c0.2\u20130.3, indicating that the title compound could be a suitable candidate as the emitting material in organic light-emitting diode (OLED) applications.The title compound, [Pt(C The average length [1.949\u2005(4)\u2005\u00c5] of the Pt\u2014C bonds is slightly shorter than that [2.042\u2005(3)\u2005\u00c5] of the Pt\u2014N bonds because of back bonding between the metal and the anionic C atom of the ligand. The Cl1 and Cl2 atoms deviate from the mean plane consisting of the Pt and coordinated N/C atoms [r.m.s. deviations = 0.013\u2005(1) (A) and 0.017\u2005(1)\u2005\u00c5 (B)] with deviations of 0.700\u2005(6)\u2005\u00c5 for A and 0.720\u2005(6)\u2005\u00c5 for B.The asymmetric unit of the title compound, Pt\u2005\u00c5, Cl2\u22efCg8 = 3.455\u2005(2)\u2005\u00c5; green dashed lines in Fig.\u00a01Cg4 and Cg8 are the centroids of the N3/C17\u2013C21 and N6/C38\u2013C42 rings, respectively] between the coordinated chloride ion and the pyridine ring with fluorine substituents are also observed. Mol\u00adecules A and B are inter\u00adlinked by a C\u2014H\u22efCl inter\u00adaction (Table\u00a02A and 9.64\u2005(11)\u00b0 for B. However, the terminal di\u00adfluoro-pyridine ring is tilted by 46.08\u2005(9) for A and 46.96\u2005(8)\u00b0 for B with respect to phenyl\u00adpyridine ring plane. This distortion may be caused by the intra\u00admolecular Cl\u22ef\u03c0 inter\u00adaction described above. The pyridine ring of the pyridine-2-yl\u00adoxy group is slightly tilted by 22.09\u2005(13) for A and 19.70\u2005(13)\u00b0 for B relative to the phenyl\u00adpyridine ring plane.In each mol\u00adecule, there are intra\u00admolecular C\u2014H\u22efCl/F inter\u00adactions, contributing to the stabilization of the mol\u00adecular structure Table\u00a02. Moreoven Table\u00a02. In the A and B mol\u00adecules and between pairs of inversion-related B mol\u00adecules lead to the formation of a two-dimensional supra\u00admolecular network lying parallel to the ab plane. In addition, this network is consolidated by halogen\u22ef\u03c0 and weak \u03c0\u2013\u03c0 stacking inter\u00adactions between pairs of inversion-related B mol\u00adecules. These sheets are stacked along the c-axis direction and connected by F\u22ef\u03c0 and weak \u03c0\u2013\u03c0 stacking inter\u00adactions between pairs of inversion-related A mol\u00adecules, resulting in the formation of a three-dimensional supra\u00admolecular network.In the crystal structure, inter\u00admolecular C\u2013H\u22efCl/F hydrogen bonds Table\u00a02 and 2 \u25b8 The bright blueish-green emission of the title compound in solution is dominated by phospho\u00adrescence as supported by an excited-state lifetime of more than 1\u2005ms. Emission maxima appear at 517 and 544\u2005nm at room temperature, as shown in Fig.\u00a04SciFinder , gave two hits. These are the reports of the crystal structures and photophysical properties of the free ligands for 2\u2032,6\u2032-di\u00adfluoro-6-[3-(pyridin-2-yl\u00adoxy)phen\u00adyl]-2,3\u2032-bi\u00adpyridine and 2\u2032,6\u2032-dimeth\u00adoxy-6-[3-(pyridin-2-yl\u00adoxy)phen\u00adyl]-2,3\u2032-bi\u00adpyridine phen\u00adyl]-2,3\u2032-bi\u00adpyridine 2, 2 and xylene (10\u2005ml) was refluxed (433\u2005K) for 48\u2005h under an N2 flow. The xylene was removed by distillation and the crude product was purified by silica gel column chromatography to give the title compound as a yellow solid in 40% yield. Orange\u2013red crystals suitable for X-ray crystallography analysis were obtained from a CH2Cl2/hexane solution by slow evaporation. 1H NMR \u03b4 9.91 , 8.20, 7.97 , 7.90\u20137.84 , 7.50\u20137.42 , 7.30\u20137.22 , 7.06 , 6.97-6.91 . 13C NMR \u03b4 206.5, 167.3, 156.4, 155.9, 151.0, 148.1, 146.6, 146.5, 140.1, 139.0, 125.7, 125.0, 121.0, 118.4, 118.3, 117.6, 115.9, 106.0, 105.9, 105.7, 105.6. Analysis calculated for C21H12ClF2N3OPt: C 42.69; H 2.05; N 7.11%; found: C 42.70, H 2.06, N 7.09%.The title compound was synthesized as follows: A mixture of the ligand , PtClUiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021000128/hb7960sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989021000128/hb7960Isup2.hklStructure factors: contains datablock(s) I. DOI: 2053861CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-019-47062-2, published online 22 July 2019Correction to: This Article contains errors in the Results section where,\u201cIt was noted that the vessel diameter of the untreated veins was (on average?) 132% higher than that of the treated veins , indicating that the treated veins were thinner.\u201dshould read:\u201cIt was noted that the vessel diameter of the untreated veins was 132% higher than that of the treated veins , indicating that the treated veins were thinner.\u201d\u201cAdditionally, the (average?) RNV vessel diameter was estimated to be 29.83\u2009\u00b1\u20094.17\u2009\u00b5m, which was 97.77% thinner than choroidal vessels (59.00\u2009\u00b1\u200911.47\u2009\u00b5m), p\u2009<\u20090.05.\u201dshould read:\u201cAdditionally, the RNV vessel diameter was estimated to be 29.83\u2009\u00b1\u20094.17\u2009\u00b5m, which was 97.77% thinner than choroidal vessels (59.00\u2009\u00b1\u200911.47\u2009\u00b5m), p\u2009<\u20090.05.\u201d"} +{"text": "II ion is coordinated in a distorted octa\u00adhedral [FeN4O2] environment by two di\u00adcyano\u00adaurate anions, two water mol\u00adecules and two partially protonated 1,2-di(4-pyrid\u00adyl)ethyl\u00adene mol\u00adecules. Di\u00adcyano\u00adaurate anions bridge the FeII cations, forming infinite chains, which propagate along the a-axis direction. The chains are connected via aurophillic inter\u00adactions with two non-coordinated di\u00adcyano\u00adaurate anions for each FeII ion.In the title compound, the Fe 2][Au(CN)2]2\u00b7bpe\u00b72H2O}n , the FeII ion is coordinated in a distorted octa\u00adhedral [FeN4O2] environment by two di\u00adcyano\u00adaurate anions, two water mol\u00adecules and two partially protonated 1,2-di(4-pyrid\u00adyl)ethyl\u00adene mol\u00adecules. Di\u00adcyano\u00adaurate anions bridge the FeII cations, forming infinite chains, which propagate along the a-axis direction. The chains are connected via aurophilic inter\u00adactions with two non-coordinated di\u00adcyano\u00adaurate anions for each FeII ion. The polymeric chains inter\u00adact with each other via \u03c0\u2013\u03c0 stacking between the guest bpe mol\u00adecules and multiple hydrogen bonds.In the title compound [Fe(bpe)(Hbpe)Au(CN) These layers are supported by N-donor aromatic ligands . Di-, tetra- and octa\u00adcyano\u00admetallic anions have been introduced to develop coordination compounds of this kind. It has been shown that the inclusion of guest mol\u00adecules can significantly influence the temperature, completeness and step character of spin transition in complexes belonging to this class (Hbpe)Au(CN)2](Au(CN)2)2\u00b7bpe\u00b72H2O in which the FeII ions are stabilized in the high-spin (HS) state.Here we describe the crystal structure of a new cyano\u00admetallic FeP4O2] octa\u00adhedral environment \u2005\u00c5, N5\u2014H5A\u22efN5i = 176\u00b0; Table\u00a01The title compound crystallizes in the triclinic nt Fig.\u00a01 formed bII coordination environment is \u03a3|90\u00a0\u2212\u00a0 \u03b8| = 6.8\u00b0, where \u03b8 are cis-N\u2014Fe\u2014N or cis-N\u2014Fe\u2014O angles. Two CN\u2212 anions bridge the Fe2+ and Au+ cations [Fe1\u22efAu1 = 5.280\u2005(3)\u2005\u00c5], creating a one-dimensional polymer, Fe1\u2014N1\u2014C1 = 172.8\u2005(7)\u00b0, N1\u2014C1\u2014Au1 = 179.1\u2005(8)\u00b0 and C1\u2014Au1\u2014C1 = 180.0\u00b0, leading to a very slight deviation from linearity of the chains. This chain binds one guest bpe and two guest water mol\u00adecules per FeII centre.The deviation from the ideal octa\u00adhedral geometry of the Fe2]\u2212 counter-ions are connected with the polymeric chains by aurophilic inter\u00adactions and C7\u22efN2B hydrogen bonds . These free counter-ions are disordered over two positions with Au1\u2014Au2A = 3.324\u2005(1)\u2005\u00c5 and Au1\u2014Au2B = 3.101\u2005(5)\u2005\u00c5. The polymeric chains are connected to each other via \u03c0\u2013\u03c0 inter\u00adactions \u2005\u00c5, \u03b1 = 10.3\u00b0, offset = 1.043\u2005\u00c5, where Cg1 and Cg2 are the centroids of the N4/C4\u2013C8 and N6/C16\u2013C20 rings, respectively, and Cg3\u22efCg4 = 3.794\u2005(6)\u2005\u00c5, \u03b1 = 6.8\u00b0, offset = 1.835\u2005\u00c5, where Cg3 and Cg4 are the centroids of N5/C11\u2013C15 and N6ii/ Cii16\u2013C20 rings, respectively]. Guest bpe mol\u00adecules are additionally linked to the polymeric chains by hydrogen bonds with coordinated water mol\u00adecules . One of the guest water mol\u00adecules forms hydrogen bonds with the coord\u00adinated water [Fig.\u00a03A\u22efO2 = 156\u00b0] and weak hydrogen bonds with free di\u00adcyano\u00adaurate counter-ions and free di\u00adcyano\u00adaurate counter-ions . Hydrogen-bonding parameters are summarized in Table\u00a01The structure is characterized by the presence of several different kinds of weak inter\u00adactions that create a three-dimensional supra\u00admolecular framework. Two free \u00adbis\u00ad(2-phenyl\u00adpyrazine)\u00adbis\u00ad(aqua)\u00adiron(II)gold(I) bis\u00ad(cyano)\u00adgold(I)] in water (0.5\u2005ml), the second was a mixture of water/ethanol and the third layer was a solution of 1,2-di(4-pyrid\u00adyl)ethyl\u00adene (0.05\u2005mmol) and [Fe(OTs)2]\u00b76H2O in ethanol (0.5\u2005ml) with 0.2\u2005ml of water. After two weeks, red crystals grew in the second layer; the crystals were collected and kept in the mother solution prior to measurement.Crystals of the title compound were prepared by the slow diffusion method between three layers in a 3\u2005ml tube. The first layer was a solution of K[Au(CN)iUso(H) = 1.2Uiso(C), Uiso(H) = 1.2Uiso(N), Uiso(H) = 1.5Uiso(O). Uaniso values for all C and N atoms in the guest di\u00adcyano\u00adaurate anions and the O2 and O3 water mol\u00adecules were constrained to be equal using the EADP command. Distances N3A\u2014C3A and N2A\u2014C2A were restrained to a target of 1.15\u2005\u00c5 and distances Au2A\u2014C3A and Au2A\u2014C2A were restrained to a target of 1.99\u2005\u00c5 using the DFIX command. The following distances were restrained to be equal using the SADI command: C2A\u2014N2A and C2B\u2014N2B; Au1\u2014C2A and Au1\u2014C2B; C3A\u2014N3A and C3B\u2014N3B; Au1\u2014C3A and Au1\u2014C3B; C2A\u2014C3A and C2B\u2014C3B.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020006738/tx2022sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020006738/tx2022Isup2.hklStructure factors: contains datablock(s) I. DOI: 2004716CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dihedral angle between the mean plane of the anthra\u00adquinone ring system and the dioxepine ring in the title compound is 16.29\u2005(8)\u00b0. The packing is consolidated by C\u2014H\u22efO, \u03c0\u2013\u03c0 and C=O\u22ef\u03c0 inter\u00adactions. 17H12O4, was synthesized from the dye alizarin. The dihedral angle between the mean plane of the anthra\u00adquinone ring system (r.m.s. deviation = 0.039\u2005\u00c5) and the dioxepine ring is 16.29\u2005(8)\u00b0. In the crystal, the mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, forming sheets lying parallel to the ab plane. The sheets are connected through \u03c0\u2013\u03c0 and C=O\u22ef\u03c0 inter\u00adactions to generate a three-dimensional supra\u00admolecular network. Hirshfeld surface analysis was used to investigate inter\u00admolecular inter\u00adactions in the solid-state: the most important contributions are from H\u22efH (43.0%), H\u22efO/O\u22efH (27%), H\u22efC/C\u22efH (13.8%) and C\u22efC (12.4%) contacts.The title compound, C The colour of anthra\u00adquinone-based compounds can be modified by the type and position of the substituents attached to the anthra\u00adquinone nucleus P21/n with one mol\u00adecule in the asymmetric unit: it consists of three fused six-membered rings and one seven-membered ring as shown in Fig.\u00a01Compound (I)q2 = 0.896\u2005(2)\u2005\u00c5, \u03c62 = 113.50\u2005(12)\u00b0, q3 = 0.358\u2005(2)\u2005\u00c5, and \u03c63 = 217.8\u2005(3)\u00b0. These metrics indicate that the ring adopts a screw boat conformation. The C\u2014O and C=O bond lengths lie within the ranges 1.355\u2005(2)\u20131.457\u2005(2)\u2005\u00c5 and 1.216\u2005(2)\u20131.226\u2005(2)\u2005\u00c5, respectively, confirming their single and double-bond character.A puckering analysis of the seven-membered ring yielded the parameters B\u22efO1 hydrogen bonds form inversion dimers with an A\u22efO3 contacts, thereby generating corrugated chains of mol\u00adecules . A C17\u2014H17B\u22efO2 hydrogen bond links the chains together (Table\u00a01b and 2c), forming sheets propagating in the ab plane. These sheets are supported by extensive \u03c0\u2013\u03c0 contacts between adjacent rings, with centroid-to-centroid distances Cg1\u22efCg2 = 3.599\u2005(2) and Cg2\u22efCg3 = 3.683\u2005(2)\u2005\u00c5 and weak C12=O1\u22ef\u03c0 [oxygen\u2013centroid distance = 3.734\u2005(2)\u2005\u00c5] inter\u00adactions es Fig.\u00a02a. A C17r Table\u00a01b and 2cns Fig.\u00a03, linkinget al., 2016viz. 1-hy\u00addroxy-2-meth\u00adoxy-6-methyl and two-dimensional fingerprint plots shows the mol\u00adecular electrostatic potential surface generated using TONTO with a STO-3G basis set in the range \u22120.050 to 0.050 a.u. within the Hartree\u2013Fock level of theory. Mol\u00adecular sites evidenced in red correspond to positive potential energy and in blue to negative potential energy de and di diagonal axes. The most important inter\u00adaction is H\u22efH, contributing 43% to the overall crystal packing, which is reflected in Fig.\u00a05b as widely scattered points of high density due to the large hydrogen content of the mol\u00adecule, with small split tips at de \u2243 di \u2243 1.2\u2005\u00c5. The contribution from the O\u22efH/H\u22efO contacts (27%) [note that the O\u22efH interactions make a larger contribution (14.6%) than the H\u22efO interactions (12.4%)], corresponding to C\u2014H\u22efO inter\u00adactions, is represented by a pair of sharp spikes characteristic of a strong hydrogen-bond inter\u00adaction, de + di \u2243 2.35\u2005\u00c5 . The significant contribution from C\u22efH/H\u22efC contacts (13.8%) to the Hirshfeld surface of (I)d, with de + di \u22432.55\u2005\u00c5. The distribution of points in the de = di \u2243 1.75\u2005\u00c5 range in the fingerprint plot delineated into C\u22efC contacts indicates the existence of weak \u03c0\u2013\u03c0 stacking inter\u00adactions between the central anthracene ring and the C6\u2013C11 and C1\u2013C4/C13\u2013C14 rings . Aromatic \u03c0\u2013\u03c0 inter\u00adactions are indicated by adjacent red and blue triangles in the shape-index map and also by the flat region around these rings in the Hirshfeld surfaces mapped over curvedness in Fig. S1c.As illustrated in Fig.\u00a05\u2005\u00c5 Fig.\u00a05c. The sgs Fig.\u00a04b and 5ede + di \u2243 3.2\u2005\u00c5 in Fig.\u00a05f.The contribution of 3.2% from C\u22efO/O\u22efC contacts is due to the presence of short inter\u00adatomic C=O\u22ef\u03c0 contacts, and is apparent as the pair of parabolic tips at N hydro\u00adchloric acid, extracted with chloro\u00adform (3 \u00d7 30\u2005ml) and then chromatographed on a silica gel column with di\u00adchloro\u00admethane/petroleum ether (1/1) as eluent, which yielded 200\u2005mg (35%) of 1,2-propyl\u00adene\u00addioxy\u00adanthra\u00adquinone as a yellow compound was treated with 1,3-di\u00adbromo-propane in di\u00admethyl\u00adformamide (30\u2005ml) in the presence of anhydrous potassium carbonate with stirring and heated to 393\u2005K for 24\u2005h. The reaction mixture was evaporated to dryness under vacuum and the resulting crude product was acidified with 12\u2005nd Fig.\u00a06. Colourl1H NMR : \u03b4 (ppm): 8.21 , 7.95 , 7.72 , 7.26 , 4.48 , 4.43 , 2.34 ; 13C NMR : \u03b4 (ppm): 182.9, 182.5, 157.3, 151.3, 135.2, 133.9, 133.4, 132.6, 129.6, 127.1, 126.5, 126.0, 125.9, 123.3, 70.5, 70.2, 30.0. Analysis calculated for C17H12O4: C, 72.85%; H, 4.32%; found: C, 72.82%; H, 4.29%.Uiso(H) = 1.2Ueq(C). The reflection (011), affected by the beam-stop, was removed during refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020003965/hb7899sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020003965/hb7899Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020003965/hb7899Isup3.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020003965/hb7899sup4.docxSupplementary figures: Hirshfeld surface analysis. DOI: 1991271CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal of the title compound, the packing is driven by O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efN hydrogen-bond inter\u00adactions along with \u03c0\u2013\u03c0 stacking inter\u00adactions. 10H8N8\u00b72H2O or H2bmtz\u00b72H2O , the asymmetric unit consists of one-half mol\u00adecule of H2bmtz and one water mol\u00adecule, the whole H2bmtz mol\u00adecule being generated by a crystallographic twofold rotation axis passing through the middle point of the 1,4-di\u00adhydro-1,2,4,5-tetra\u00adzine moiety. In the crystal, N\u2014H\u22efO, N\u2014H\u22efN, O\u2014H\u22efO hydrogen bonds and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions link the components into a three-dimensional supra\u00admolecular network. Hirshfeld surface analysis was used to further investigate the inter\u00admolecular inter\u00adactions in the crystal structure.In the title compound, C Many compounds of this class are bioactive (Jubeen I) is shown in Fig.\u00a012bmtz and one water mol\u00adecule, in which the whole mol\u00adecule of the H2bmtz is generated by a crystallographic twofold rotation axis passing through the middle point of the 1,4-di\u00adhydro-1,2,4,5-tetra\u00adzine moiety. The H2bmtz mol\u00adecule is therefore not planar (r.m.s. deviation from planarity = 0.598\u2005\u00c5) with a C4\u2014C5\u2014N3\u2014N4i torsion angle of 178.46\u2005(14)\u00b0 . The pyrimidine rings are twisted with respect to each other, making a dihedral angle of 43.67\u2005(9)\u00b0. The 1,4-di\u00adhydro-1,2,4,5-tetra\u00adzine moiety adopts a twist-boat conformation with a C5\u2014N3\u2014N4i\u2014C5i torsion angle of \u221241.17\u2005(17)\u00b0. The N3\u2014N4i and C5\u2014N4 bond lengths of 1.423\u2005(2) and 1.395\u2005(2)\u2005\u00c5, confirm their single-bond character, while the C3\u2014N5 bond length of 1.278\u2005(2)\u2005\u00c5, is consistent with a double bond \u2005\u00c5]. At the same time, the water mol\u00adecules are connected by O\u2014H\u22efO hydrogen bonds make a less significant contribution. The contribution of the H\u22efO/O\u22efH (9.0%) contacts (i.e. C\u2014H\u22efO and O\u2014H\u22efO) and other contacts such as C\u22efC (7.1%) (i.e. \u03c0\u2013\u03c0 stacking), H\u22efC/C\u22efH (6.1%) and N\u22efN (4.7%) make a small contribution to the entire Hirshfeld surface.To further qu\u00adantify the nature of the inter\u00admolecular inter\u00adactions present in the crystal structure, Hirshfeld surfaces -1,2,4,5-tetra\u00adzine]bis\u00ad[bis\u00ad(tri\u00adphenyl\u00adphosphine)cop\u00adper(I)] bis\u00ad(tetra\u00adfluorido\u00adborate) di\u00adchloro\u00admethane solvate -1,4-di\u00adhydro-1,2,4,5-tetra\u00adzine][\u03bc2-(di\u00adcyano\u00adethen\u00adyl\u00ad\u00adidene)amido][(di\u00adcyano\u00adethenyl\u00adidene)amido]\u00adaceto\u00adnitrile\u00addisilver(I)] .All commercially available chemicals and solvents were of reagent grade and were used as received without further purification. HUiso(H) = 1.2Ueq(C). The 1,4-di\u00adhydro-1,2,4,5-tetra\u00adzine and water H atoms were located in difference-Fourier maps and were constrained to N\u2014H = 0.86 \u00b1 0.01\u2005\u00c5 with Uiso(H) = 1.2Ueq(N) and O\u2014H = 0.84 \u00b1 0.01\u2005\u00c5 with Uiso(H) = 1.5Ueq(O), respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020002765/hb7895sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020002765/hb7895Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020002765/hb7895Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020002765/hb7895Isup4.cmlSupporting information file. DOI: 1986751CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the title compound contains two independent mol\u00adecules, consisting of perimidine and phenol units, which are linked through an N\u2014H\u22efO hydrogen bond. Intra\u00admolecular O\u2014H\u22efN hydrogen bonds are observed in both independent mol\u00adecules. 17H14N2O, contains two independent mol\u00adecules each consisting of perimidine and phenol units. The tricyclic perimidine units contain naphthalene ring systems and non-planar C4N2 rings adopting envelope conformations with the C atoms of the NCN groups hinged by 44.11\u2005(7) and 48.50\u2005(6)\u00b0 with respect to the best planes of the other five atoms. Intra\u00admolecular O\u2014H\u22efN hydrogen bonds may help to consolidate the mol\u00adecular conformations. The two independent mol\u00adecules are linked through an N\u2014H\u22efO hydrogen bond. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (52.9%) and H\u22efC/C\u22efH (39.5%) inter\u00adactions. Hydrogen bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Density functional theory (DFT) optimized structures at the B3LYP/ 6\u2013311\u2005G level are compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.The asymmetric unit of the title compound, C H Perimidines are defined as peri-naphtho-fused pyrimidines , carried out at the B3LYP/6-311G level, are compared with the experimentally determined mol\u00adecular structure in the solid state.I, contains two crystallographically independent mol\u00adecules each consisting of perimidine and phenol units, where the tricyclic perimidine units contain naphthalene ring systems and non-planar C4N2 rings and B\u2032 (N1A/N2A/C1A/C9B\u2013C11B) rings gave the parameters q2 = 0.9280\u2005(12)\u2005\u00c5, q3 = 0.1829\u2005(12)\u2005\u00c5, QT = 0.9459\u2005(13)\u2005\u00c5, \u03b82 = 75.85\u2005(15)\u00b0 and \u03c6 2= 134.47\u2005(18)\u00b0 for B and q2 = 0.5320\u2005(11)\u2005\u00c5, q3 = 0.3791\u2005(11)\u2005\u00c5, QT = 0.6533\u2005(14)\u2005\u00c5, \u03b82 = 54.33\u2005(12)\u00b0 and \u03c6 2= \u22125.47\u2005(13)\u00b0 for B\u2032; both rings adopt envelope conformations, where atoms C1A and C1B are at the flap positions and at distances of 0.6044\u2005(12) and \u22120.6590\u2005(13)\u2005\u00c5, respectively, from the best planes through the other five atoms. The C4N2 rings may alternatively be described as being hinged about the N\u22efN vectors with the N1A/C1A/N2A and N1B/C1B/N2B planes being inclined by 44.11\u2005(7) and 48.50\u2005(6)\u00b0, respectively, to the best planes through the other five atoms (N1A/N2A/C9A\u2013C11A) and (N1B/N2B/C9B\u2013C11B). Rings A (C2A\u2013C7A), C (C10A\u2013C15A), D (C9A/C10A/C15A\u2013C18A) and A\u2032 (C2B\u2013C7B), C\u2032 (C10B\u2013C15B), D\u2032 (C9B/C10B/C15B\u2013C18B) are oriented at dihedral angles of A/C = 76.78\u2005(4), A/D = 78.49\u2005(4), C/D = 2.09\u2005(3)\u00b0 and A\u2032/C\u2032 = 88.43\u2005(3), A\u2032/D\u2032 = 88.31\u2005(3), C\u2032/D\u2032 = 3.26\u2005(4)\u00b0. Intra\u00admolecular O\u2014H\u22efN hydrogen bonds analysis have the tips at de + di = 2.50\u2005\u00c5. The scattered points in the pair of spikes in the fingerprint plot delineated into H\u22efO/O\u22efH have a symmetrical distribution with the tips at de + di = 2.49\u2005\u00c5. The H\u22efN/N\u22efH contacts have a distribution of points with the tips at de + di = 2.72\u2005\u00c5. Finally, the C\u22efC inter\u00adactions are reflected in Fig.\u00a04f as low density wings with the tips at de + di = 3.60\u2005\u00c5.The shape-index of the HS is a tool to visualize the \u03c0\u2013\u03c0 stacking by the presence of adjacent red and blue triangles; if there are no adjacent red and/or blue triangles, then there are no \u03c0\u2013\u03c0 inter\u00adactions. Fig.\u00a03dnorm plotted onto the surface are shown for the H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions in Fig.\u00a05a\u2013c, respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH and H\u22efC/C\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing using standard B3LYP functional and 6\u2013311\u2005G basis-set calculations , hardness (\u03b7), potential (\u03bc), electrophilicity (\u03c9) and softness (\u03c3) are recorded in Table\u00a03\u03c3 is for the evaluation of both the reactivity and stability. The electron transition from the HOMO to the LUMO energy level is shown in Fig.\u00a06H-perimidin-2-yl)phenol ring. The energy band gap [\u0394E = ELUMO - EHOMO] of the mol\u00adecule is 1.4933\u2005eV, the frontier mol\u00adecular orbital energies EHOMO and ELUMO being \u22123.2606 and \u22121.7673\u2005eV, respectively.The optimized structure of the title compound, I, are II phenol product (Rf = 0.70 in hexa\u00adne/ethyl acetate (1:0.5), yield: 97% A significant qu\u00adantity of the colourless monocrystalline product was obtained by the slow evaporation of the solvent after 15 days.0.35\u2005mol (1.48\u2005g) of 1,8-di\u00adaminona\u00adphthalene and 18.8\u2005mmol (2\u2005ml) of salicyl\u00adaldehyde were introduced into a 250\u2005ml flask and 30\u2005ml of ether were added thereto. The mixture was stirred magnetically for 3 days. The grey precipitate that formed was recovered by filtration, washed with ether, rinsed with ethanol and dried under B\u00fcchner. The resulting brownish powder was recrystallized several times from ethanol to obtain colourless 2- = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in are summarized in Table\u00a0410.1107/S2056989020005939/lh5957sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020005939/lh5957Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020005939/lh5957Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020005939/lh5957Isup4.cmlSupporting information file. DOI: 1976884CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The metalated species were characterized in situ by NMR spectroscopy and in two cases isolated as crystalline solids. Single\u2010crystal XRD studies revealed the presence of salt\u2010like structures with strongly interacting ions. Synthetic applications of K[(R2N)2P(BH3)] were studied in reactions with a 1,2\u2010dichlorodisilane and CS2, which afforded either mono\u2010 or difunctional phosphine boranes with a rare combination of electronegative amino and electropositive functional disilanyl groups on phosphorus, or a phosphinodithioformate. Spectroscopic studies gave a first hint that removal of the borane fragment may be feasible.Metalation of secondary diaminophosphine boranes by alkali metal amides provides a robust and selective access route to a range of metal diaminophosphide boranes M[(R It\u2032s just so plain borane: Alkali metal diaminophosphide boranes, which can be selectively prepared by metalation of PH\u2010substituted precursors with non\u2010nucleophilic amides, were for the first time isolated and characterized crystallographically. Their synthetic application as highly functional nucleophilic building blocks is illustrated in metatheses with a dichlorodisilane and the formation of a stable carbon disulfide adduct. I or diorganophosphide boranes II ,Ib,c/IIb,c)3Si, IdIb/IIb) parent (Ic/IIc) and silylated phosphides (Id) are special because their ability to undergo electrophilic post\u2010functionalization of reactive P\u2212H and P\u2212Si bonds after the initial metathesis step makes them essentially polyfunctional building blocks.Typical phosphide reagents carry usually chemically inert alkyls or aryls 2P(Fe(CO)4]KIII, R=Et, Ph) and [(Me2N)(Ph3C)P(W(CO)5]KIV) containing metal\u2010stabilized mono\u2010 or diamino\u2010phosphido ligands were generated as spectroscopically detectable entities by deprotonation of PH\u2010substituted precursor complexes and shown to react as P\u2010centered nucleophiles. The significance of diaminophosphide boranes as synthetic intermediates was first recognized by the group of Knochel,1\u2009a] that had been generated in situ by lithium reduction of a chlorophosphine borane precursor but was neither positively identified nor further characterized (Scheme\u20051\u2009a]) and lithium diethylaminophosphide borane (Li[1\u2009a]) can be alternatively accessed through metalation of a secondary diaminophosphine borane precursor 2PCl and LiBH42N)2PCl with Me2S\u22c5BH3 furnished only inseparable mixtures of 2\u2009b with Me2NH\u22c5BH3 (10\u201320\u2005mol\u2009%). As the by\u2010product did not interfere with the subsequent metalation reactions, the mixtures were used without further purification. Reactions aiming at the generation of phosphides were carried out in THF, diethyl ether, or mixtures thereof, and reaction monitoring and product identification was generally achieved in situ by multinuclear NMR spectroscopy. We found that reliable and highly selective generation of phosphide boranes M[1\u2009b\u2013f] was feasible by treating the PH\u2010substituted precursors 2\u2009b\u2013e with a slight excess of a non\u2010nucleophilic alkali amide such as lithium di(isopropyl)amide (LDA) or a metal hexamethyldisilazide (MHMDS=MN(SiMe3)2, M=Li, Na, K, Scheme\u20052\u2009b with NaHDMS in a THF/ether mixture and with KHMDS in the presence of a crown ether permitted also the isolation of first crystalline diaminophosphide boranes of composition Na[1\u2009b]\u22c5THF and K[1\u2009b] , respectively. Both products were characterized by single\u2010crystal X\u2010ray diffraction studies (see further below).To deepen our insight into the formation of diaminophosphide boranes, we tested the routes reported for accessing Li[1\u2009c] and heterocyclic derivatives like K[1\u2009d], K[1\u2009e], while selective lithiation of heterocyclic secondary diaminophosphine borane 2\u2009e was best accomplished with LDA. Metalation of the secondary diazaphospholene borane 2\u2009f turned out a borderline case. Treatment with KHMDS produced detectable amounts of the expected, highly moisture and air sensitive phosphide K[1\u2009f], which was identified by spectroscopic data and chemical trapping with a proton source. However, both deprotonation and recovery of the secondary phosphine borane were in this case unselective and produced side\u2010products that could neither be successfully identified nor separated. We relate the difficulties encountered in the generation of K[1\u2009f] to the unusual electronic structure of unprotected secondary diazaphospholenes, which display a hydride reactivity that contrasts the behavior of alkyl and arylphosphines and is associated with an \u201cUmpolung\u201d of the P\u2212H bond.Moreover, using KHMDS for deprotonation enabled both the generation of a sterically congested diaminophosphide reagent like K,1\u2009c] but proceeded unselectively and irreproducibly with heterocyclic substrates. The use of Grignard reagents was tested with sterically uncongested diaminophosphine boranes 2\u2009a,b. While MeMgCl converted both substrates cleanly into the corresponding phosphides, EtMgBr was found to be unreactive.Application of other deprotonating reagents than amides gave erratic results. Metalation with 3\u2009a,b3\u2009b, which afforded a low yield of the metalation product K[1\u2009b] along with a main product later identified4\u2009b 4\u2009b and substantial amounts of unidentified by\u2010products. We have currently no concise explanation for the varying selectivity, even if it is tempting to relate the observed changes to different sterics of the substrates.Exploring the reduction of chlorophosphine boranes 1\u2009b\u2013f] comes from 31P and 11B\u2005NMR spectra. The signals display the expected multiplet structures arising from spin\u2010coupling between 11B (I=3/2) and 31P (I=1/2) nuclei and lack a characteristic splitting due to 1JPH coupling across a P\u2212H bond. As previously noted1\u2009a] and K[1\u2009a], the 31P\u2005NMR chemical shifts of specimens with identical diaminophosphide units show metal\u2010dependent changes, and the presence of a cation sequestering reagent like dibenzo\u201018\u2010crown\u20106 induces increases in \u03b431P of potassium phosphide boranes K by 10 to 15\u2005ppm. Both effects suggest the presence of tight contact ion pairs in solution. The non\u2010uniform trends observed discredit a simplistic assumption that declining covalency in the phosphorus\u2010metal bonding with growing size of the metal ion induces a deshielding of the 31P nucleus. The 11B\u2005NMR signals of M[1\u2009a\u2013f] display positive \u201cmetalation shifts\u201d (\u0394\u03b411B +5 to +11\u2005ppm) and numerically reduced 1JP11B coupling constants relative to the corresponding secondary phosphine borane precursors 2\u2009a\u2013f. The same trends hold for dialkyl and diarylphosphide boranes and an unusually low shift for sterically congested M[1\u2009c] . A concise explanation for these effects is yet lacking.The diagnostically most valuable information for the identification of metalated diaminophosphide boranes M[ 11B I=3/ and 31P 11B I=3/ and 31P 1\u2009b] (space group P1\u203e) and monoclinic K[1\u2009b] (space group P21/c) and important metrical parameters are displayed in Figures\u2005Graphical representations of the results of single\u2010crystal X\u2010ray diffraction studies on triclinic Na(THF)\u2009b (space1\u2009b] contain dimeric units assembled from two anionic diaminophosphide borane and two cationic Na(THF) fragments to 2.39(2)\u2005\u00c5) and a weaker contact to a B\u2212H bond of the adjacent borane in the same unit (Na\u2010H 2.69(2)\u2005\u00c5). Interactions of this type have precedence in the structures of alkyl/arylphosphide boranes[1\u2009b] the dimeric units to form one\u2010dimensionally infinite strands aligned parallel to the crystallographic a\u2010axis. If we count the \u03b72\u2010bound borane as one ligand, the metal ions exhibit an irregular pseudo\u2010[4+1] coordination geometry in which the weak \u201cintramolecular\u201d B\u2212H interaction is aligned roughly opposite to the oxygen atom of the THF ligand. The phosphorus atom displays a distorted tetrahedral coordination with bond angles from 103.1(1)\u00b0 (N1\u2212P1\u2212B1) to 118.8(1)\u00b0 (Na1\u2212P1\u2212B1). The P\u2212B and P\u2212N distances are in the range of single bonds, with the deviation of 0.053(2)\u2005\u00c5 between the P\u2212N distances reflecting the different coordination numbers of the nitrogen atoms. While the Na1\u2212O1 distance of 2.311(1)\u2005\u00c5 is a close match to the sum of covalent radii (2.32(11)\u2005\u00c5Crystals of Na(THF)\u2009b contaie boranes15, 171\u2009b] contains two independent anionic units and two cations which aggregate, as in the sodium analogue, via K\u22c5\u22c5\u22c5N and K\u22c5\u22c5\u22c5P interactions to form strongly puckered six\u2010membered cyclic arrays . The coordination sphere of the linking metal ion is completed by additional contacts to a \u03b73\u2010bound borane unit (H\u22c5\u22c5\u22c5K2 2.645(27) to 2.762(31)\u2005\u00c5) from the second next dimeric unit in the row, which reinforce the supramolecular assembly and give rise to the formation of a double stranded array of heterocyclic units extending along the crystallographic b\u2010axis. Completion of the coordination sphere of the N2\u2010coordinated potassium ion is accomplished by weaker BH\u22c5\u22c5\u22c5K interactions (H\u22c5\u22c5\u22c5K1 2.69(3) to 3.04(3)\u2005\u00c5) with borane units from adjacent, parallel strands. These secondary interactions result ultimately in the formation of two\u2010dimensional layers, which extend in the crystallographic a,b\u2010plane and are held together by van\u2010der\u2010Waals interactions. The P\u2212N (1.697(2) to 1.741(2)\u2005\u00c5) and P\u2212B distances (1.935(4), 1.947(3)\u2005\u00c5 are similar as in Na(THF)[1\u2009b] and correspond to single bonds. The P\u22c5\u22c5\u22c5K (3.301(1), 3.352(1)\u2005\u00c5) and N\u22c5\u22c5\u22c5K distances (2.849(2) to 2.961(2)\u2005\u00c5) exceed the reported sum of covalent radii (P\u2212K 3.10(15), N\u2212K 2.74(13)\u2005\u00c5Crystalline K and K[1\u2009e] afforded exclusively monosubstitution products , even when the nucleophile was employed in excess or crystalline solids and characterized by analytical and spectroscopic data. The 1H, 11B, 13C and 31P\u2005NMR spectra are unremarkable and confirm the proposed constitution. The 29Si NMR spectrum of 7 is peculiar as the expected multiplet (representing the X\u2010part of an ABX spin system) degenerates accidentally to a simple doublet. One of the two expected 29Si NMR signals of 6\u2009e stayed undetected for unknown reasons.To explore the feasibility of this approach, we studied reactions of diaminophosphide boranes with commercially available 1,2\u2010dichloro\u20101,1,2,2\u2010tetramethyldisilane s Scheme\u2005. In cont6\u2009e and 7 were further confirmed by single\u2010crystal X\u2010ray diffraction studies. Crystals of 7 \u00b0 and 180\u00b0, Figures\u20056\u2009e holds likewise two independent molecules, both of which exhibit, however, a nearly eclipsed conformation around the Si\u2212Si bond. The chlorine and phosphorus atoms in one molecule adopt a gauche\u2010orientation . The N\u2010heterocyclic rings exhibit similar twist conformations as in borane\u2010free diazaphospholidines7 are unremarkable and suggest the presence of normal single bonds. The P\u2212N distances in both complexes are shorter than in free acyclic or N\u2010heterocyclic phosphines,octane (DABCO).11B\u2005NMR spectroscopy as the expected bis\u2010borane adduct of DABCO which lacks the characteristic broadening arising from spin\u2010coupling to a quadrupolar 11B nucleus and is tentatively assigned to the targeted borane\u2010free diphosphino\u2010disilane. In addition, signals attributable to unknown borane\u2010free phosphorus\u2010containing species and bis(diethylamino)phosphine were detected, and the 11B\u2005NMR spectrum revealed further the presence of residual 7 . The product distribution observed can be explained by assuming that de\u2010protection is in principle feasible but remains incomplete and is accompanied by P\u2212Si bond cleavage arising from adventitious hydrolysis. Attempts to isolate any of the de\u2010protected phosphorus\u2010containing products remained unsuccessful.The 1,2\u2010diphosphino\u2010disilane representing the backbone of 1\u2009a] with carbon dioxide and carbon disulfide. While the former gave rise to an intractable colorless solid that could as yet not be unambiguously characterized, the reaction with CS2 proceeded cleanly to afford a soluble species which was isolated as a red, air and moisture stable crystalline solid after work\u2010up and recrystallization from ether/pentane. Analytical and MS data are in accordance with the formation of phosphinodithioformate borane 8 to 3.435(2)\u2005\u00c5, K\u22c5\u22c5\u22c5O (2.820(2)\u2005\u00c5), and weaker contacts to a remote sulfur and two hydrogen atoms of a \u03b72\u2010coordinated borane unit (K\u22c5\u22c5\u22c5S 3.806(2)\u2005\u00c5, K\u22c5\u22c5\u22c5H 2.62(6), 2.85(6)\u2005\u00c5). The anion is built around a phosphorus atom with distorted tetrahedral and a carbon atom with planar coordination. The P\u2212N (1.653(4), 1.663(3)\u2005\u00c5) and P\u2212B (1.915(6)\u2005\u00c5) distances are still shorter than in 6\u2009e, 7 but remain in the range of single bonds. The P\u2212C (1.849(4)\u2005\u00c5) and C\u2212S (1.680(54), 1.687(4)\u2005\u00c5) distances match those reported for zwitterionic triaminophosphonio dithioformates (R2N)3P+\u2013CS2\u2212 (P\u2212C 1.848(4), 1.849(2)\u2005\u00c5; C\u2212S 1.651(4) to 1.675(3)\u2005\u00c52N)3P+\u2013CS2Me (P\u2212C 1.852(5)\u2005\u00c52N)2P\u2013CS2\u2013P(NR2)2 containing a tri\u2010coordinate P atom (P\u2212C 1.895(4)\u2005\u00c52 fragment are rather insensitive to the nature of the P substituents.Considering all interactions together, the potassium ions can be assigned a [5+3] coordination sphere arising from close contacts to the sulfur atoms of two \u03bcIn this work, we established the metalation of secondary diaminophosphine boranes with alkali metal amides as broadly applicable and robust access route to nucleophilic diaminophosphide building blocks based on both acyclic and N\u2010heterocyclic molecular frameworks. Hexamethyldisilazides emerged as reagents of choice because of their ability to react as universally applicable bases which allow free selection of alkali metals. Alternative approaches to the target molecules, such as metalation of the diaminophosphine boranes with organometallic reagents or reductive metalation of P\u2010chloro\u2010substituted precursors, worked well in selected cases but gave generally erratic results. Moreover, borane complexes of diazaphospholenes materialized as critical substrates, which reacted unselectively to afford low yields of spectroscopically detectable metalation products beside unidentified by\u2010products. We presume that this behavior reflects the pertinent hydride character of the P\u2212H bonds in the free phosphines.Beyond previous findings, sodium and potassium dimethylaminosphosphide boranes were for the first time isolated as room\u2010temperature\u2010stable solids. Their crystallographic characterization reveals the presence of salt\u2010like structures that are distinguished by intimate interactions between ions of opposite charge via a combination of dative M\u22c5\u22c5\u22c5N and M\u22c5\u22c5\u22c5P and agostic M\u22c5\u22c5\u22c5H\u2212B contacts. While both types of interactions are not unprecedented for aryl and alkyl phosphide boranes, the availability of additional nitrogen donor centers increases the density of the interaction network.Last, but not least, the potential of diaminophosphide boranes to act as synthetic building blocks is illustrated by their conversion into phosphine boranes that are distinguished by a combination of electronegative amino and electropositive functional silyl groups. Initial studies indicate that removal of the borane units is in principle feasible, and we are currently striving to develop reliable synthetic protocols to accomplish this task.2O, toluene) or by using a solvent purification system . NMR spectra were recorded on Bruker Avance 250 or Avance 400 instruments. Chemical shifts in 1H\u2005NMR spectra were referenced to TMS using the residual signals of the deuterated solvent =1.73\u2005ppm; \u03b4(C6D6)=7.15\u2005ppm) as secondary reference. NMR Spectra of heteronuclei were referenced using the \u039e\u2010scale\u039e=25.145020\u2005MHz, 13C; \u039e=19.867187\u2005MHz, 29Si), BF3Et2O and 85\u2009% H3PO4 as secondary references. Coupling constants involving boron refer to 11B isotopomers, with 1:1:1:1 multiplets being denoted as q. The product composition in metalation experiments was derived from analysis of the 1H\u2005NMR signals of the N\u2010substituents of 1\u2009b\u2013f. Elemental analyses were carried out with an Elementar Micro Cube elemental analyzer. Mass spectra were obtained with a Bruker Daltonics Mikrotof\u2010Q\u2010Mass spectrometer. Phosphine boranes 2\u2009a,2\u2009e3\u2009aAll reactions were carried out under an atmosphere of inert Argon and in flame\u2010dried glassware if not mentioned otherwise. Solvents were dried by distillation from alkali metals at T=130(2) K. The structures were solved by direct methods [1\u2009b], K[1\u2009b], 6\u2009e, 7, 8) contain the supplementary crystallographic data for this paper. These data are provided free of charge by the joint Cambridge Crystallographic Data Centre and Fachinformationszentrum Karlsruhe Access Structures service.Deposition numbers\u2005Bis(dimethylamino)phosphine borane (2\u2009b): Method A: PCl3 (0.1\u2005mL 1.14\u2005mmol) was added under stirring to a cooled (\u221278\u2009\u00b0C) solution of P(NMe2)3 (0.42\u2005mL 2.29\u2005mmol) in Et2O (50\u2005mL). Stirring was continued while the solution was allowed to warm to ambient temperature. The solution was cooled again to \u221278\u2009\u00b0C and BH3\u22c5SMe2 was added. The mixture was stirred for 1\u2005h at \u221278\u2009\u00b0C and allowed to warm to RT. After cooling once more to \u221278\u2009\u00b0C, LiAlH4 was added. The mixture was stirred for 1\u2005h at \u221278\u2009\u00b0C, allowed to warm to RT, and stirred for further 24\u2005h. Water (0.5\u2005mL) was added and volatiles were evaporated under reduced pressure. The solid residue was extracted with hexane (20\u2005mL). The extract was dried (Na2SO4) and solids were removed by filtration. Evaporation of the solvent under reduced pressure afforded 2\u2009b as colorless solid which melted around RT . Method B: PCl3 (0.49\u2005mL 5.6\u2005mmol), P(NMe2)3 (2.00\u2005mL 11.2\u2005mmol) and BH3\u22c5SMe2 were reacted in Et2O (30\u2005mL) as described under A. Li[BEt3H] was added. The mixture was allowed to warm to RT and stirred for further 18\u2005h. Volatiles were evaporated under reduced pressure and the residue was taken up in pentane (150\u2005mL). The resulting mixture was filtered over Celite. Evaporation of the solvent under reduced pressure gave 2\u2009b as colorless oil , which contained according to NMR measurements 10 to 20\u2009% dimethylamine borane. 31P{1H} NMR (C6D6): \u03b4=89.2\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221240.2\u2005ppm ; 1H\u2005NMR (C6D6): \u03b4=5.96 , 2.27 , 1.42\u2005ppm ; 13C{1H} NMR (C6D6): \u03b4=38.5\u2005ppm ; MS(ESI): m/z=119.08 .Bis(diisopropylamino)phosphine borane (2\u2009c): LiBH4 was added to a stirred solution of (iPr2N)2PCl in Et2O (25\u2005mL). The mixture was stirred for 1\u2005h. Volatiles were evaporated under reduced pressure and the residue extracted with hexane (4\u00d720\u2005mL). The combined extracts were filtered, the filtrate concentrated under reduced pressure, and the product isolated by crystallization at \u221224\u2009\u00b0C . 31P{1H} NMR (C6D6): \u03b4=46.3\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221235.6\u2005ppm ; 1H\u2005NMR (C6D6): \u03b4=6.70 , 3.43 , 1.61 , 1.10 , 0.98\u2005ppm ; 13C{1H} NMR (C6D6): \u03b4=46.4\u2005ppm , 22.6 ; MS(ESI): m/z=246.2 [M+]; C12H32BN2P (246.19\u2005g\u2009mol\u22121): calcd C 58.55 H 13.10 N 11.38, found C 58.28 H 12.84 N 11.13.1,3\u2010Bis\u20101,3,2\u2010diazaphospholidine borane (2\u2009d): NaBH4 was added under stirring to a cooled (0\u2009\u00b0C) solution of 2\u2010chloro\u20101,3\u2010bis1,3,2\u2010diazaphospholidine in MeCN (10\u2005mL). The mixture was stirred for 24\u2005h and during this time allowed to warm slowly to RT. Water (0.5\u2005mL) was added, volatiles evaporated under reduced pressure and the residue extracted with toluene (75\u2005mL). The extract was dried (Na2SO4) and filtered. Evaporation of the filtrated gave 2\u2009d as yellow solid . 31P{1H} NMR (C6D6): \u03b4=75.3\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221239.8\u2005ppm ; 1H\u2005NMR (C6D6): \u03b4=7.35 , 6.98 , 6.91 , 3.21\u20132.88 , 2.48 , 2.10 , 1.2\u2005ppm ; 13C{1H} NMR (C6D6): \u03b4=139.3 , 138.0 , 129.1 , 128.6 , 49.1 , 18.7 , 18.5\u2005ppm ; MS ((+)\u2010ESI): m/z=337.14 [MK+\u2013BH3]; C18H26BN2P (312.19\u2005g\u2009mol\u22121): calcd C 69.25 H 8.39 N 8.97, found C 69.00 H 8.51 N 8.68.1,3\u2010Bis\u20101,3,2\u2010diazaphospholene borane (2\u2009f): NaBH4 was added to a suspension of 2\u2010bromo\u20101,3\u2010bis\u20101,3,2\u2010diazaphospholene in MeCN (50\u2005mL). The mixture was stirred for 18\u2005h, filtered, and evaporated to dryness. The residue was washed with pentane and dried in vacuum to afford 477\u2005mg (77\u2009%) of crude 2\u2009f. Further purification was feasible by extracting the crude product with benzene, filtration, and evaporation of the filtrate to dryness. 31P{1H} NMR (C6D6): \u03b4=79.8\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221240.2\u2005ppm ; 1H\u2005NMR (C6D6): \u03b4=8.10 , 7.01\u20136.82 , 5.37 , 2.43 , 2.10 , 1.6\u2005ppm ; 13C{1H} (C6D6): \u03b4=138.7 , 137.4 , 136.4 , 129.0 , 128.4 , 127.8 , 118.8 , 19.3 , 18.5\u2005ppm ; MS ((+)\u2010ESI): m/z=310.19 [M+], 311.19 [M+H+]; C18H24BN2P (310.19\u2005g\u2009mol\u22121): calcd C 69.70 H 7.80 N 9.03, found C 68.45 H 7.88 N 9.06. The reason for the low carbon content is not known.Chloro\u2010bis(dimethylamino)phosphine borane (3\u2009b): BH3\u22c5SMe2 was added under stirring to a cooled (0\u2009\u00b0C) solution of (Et2N)2PCl in Et2O (10\u2005mL). After 30\u2005min, the solution was allowed to warm to RT and stirred for further 18\u2005h. The solvent was then removed under reduced pressure and the residue dissolved in hexane (10\u2005mL). After reaction control by 31P\u2005NMR, enough BH3\u22c5SMe2 to convert unreacted starting material and more hexane (10\u2005mL) were added. Filtering the mixture over Celite and removal of the solvent under reduced pressure gave 3\u2009b as yellow oil . The product is thermally unstable and decomposes even below room temperature; it must be used immediately and cannot be stored.31P{1H} NMR (C6D6): \u03b4=139.7\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221236.0\u2005ppm ; 1H\u2005NMR (C6D6): \u03b4=2.27 , 1.59\u2005ppm ; 13C{1H} (C6D6): \u03b4=36.7\u2005ppm . The thermal instability precluded obtaining a meaningful elemental analysis.General procedure for the metalation of secondary diaminophosphine boranes 2\u2009b,d,f with alkali metal bis(trimethylsilyl)amides: MHMDS was added to a 0.06\u20130.2\u2009m solution of the secondary diaminophosphine borane in THF or [D8]THF (1 to 2\u2005mL). The solution was stirred for 10\u2005min at RT. An aliquot of the reaction mixture was then transferred to an NMR tube and characterized by multinuclear NMR spectroscopy. Conversion to the metalation product was derived from evaluation of the integrals in 1H\u2005NMR spectra. Deprotonation at phosphorus was in all cases confirmed by the disappearance of the characteristic signal splitting due to 1JPH.1\u2009b]: Conversion 86\u2009%. 31P{1H} NMR (C6D6): \u03b4=145.9\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221234.6\u2005ppm ; 1H\u2005NMR ([D8]THF): \u03b4=2.73 , 0.64\u2005ppm ; 13C{1H} NMR ([D8]THF): \u03b4=43.8\u2005ppm .Li[1\u2009b]: Conversion 90\u2009%. 31P{1H} NMR (C6D6): \u03b4=145.0\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221234.3\u2005ppm ; 1H\u2005NMR ([D8]THF): \u03b4=2.76 , 0.66\u2005ppm ; 13C{1H} NMR ([D8]THF): \u03b4=44.9\u2005ppm .Na[1\u2009b]: Conversion 84\u2009%. 31P{1H} NMR (C6D6): \u03b4=142.9\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221232.9\u2005ppm ; 1H\u2005NMR ([D8]THF): \u03b4=2.7 , 0.70\u2005ppm ; 13C{1H} NMR ([D8]THF): \u03b4=44.3\u2005ppm .K[1\u2009f]: conversion 70\u2009%. 31P{1H} NMR (C6D6): \u03b4=201.6\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221229.5\u2005ppm ; 1H\u2005NMR ([D8]THF): \u03b4=7.00\u20136.63 , 5.54 , 2.46\u2005ppm ; 13C{1H} NMR ([D8]THF): \u03b4=127.6 , 121.9 , 127.8 , 117.6 , 19.5\u2005ppm .K[Isolation of sodium bis(dimethylamino)phosphide borane Na[1\u2009b]: A solution of Na[1\u2009b] was prepared as described above from NaHMDS and 2\u2009b in THF (2\u2005mL). Storage at \u221224\u2009\u00b0C produced colorless, highly air and moisture sensitive crystals, which were separated by decantation (no yield determined) and characterized by NMR spectroscopy (see Table\u2005Isolation of potassium bis(dimethylamino)phosphine borane K[1\u2009b]: KHMDS and dibenzo\u201018\u2010crown\u20106 were added to a solution of 2\u2009b in a mixture of Et2O (3\u2005mL) and THF (7\u2005mL) was carried out as described above. The mixture was stirred for 5\u2005min. Storage at \u221224\u2009\u00b0C produced colorless, highly air and moisture sensitive crystals, which were separated by decantation (no yield determined) and characterized by NMR spectroscopy was evaporated to dryness. A solution of 2\u2009b in [D8]THF (1\u2005mL) was added. The solution was stirred for 5\u2005min and then analyzed by multinuclear NMR spectroscopy : \u03b4=142.4\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221233.0\u2005ppm ; 1H\u2005NMR ([D8]THF): \u03b4=2.58 , 0.45\u2005ppm ; 13C{1H} NMR ([D8]THF): \u03b4=43.4\u2005ppm .General procedure for the metalation of secondary diaminophosphine boranes 2\u2009c,e: The metalation agent ][1\u2009c]: 1.2\u2005equiv. KHMDS and 1.0\u2005equiv. dibenzo\u201018\u2010crown\u20106; for Li[1\u2009e]: 1.33\u2005equiv. LDA; for [K(dibenzo\u201018\u2010crown\u20106)][1\u2009e]: 2.0\u2005equiv. KHMDS and 1.0\u2005equiv. dibenzo\u201018\u2010crown\u20106) was added to a 3\u20136\u2005mm solution of the secondary diaminophosphine borane in Et2O (20 to 25\u2005mL). After stirring for 1\u2005h, volatiles were removed under reduced pressure. The residue was dissolved in [D8]THF or C6D6 and characterized by multinuclear NMR spectroscopy. 31P and 11B\u2005NMR data are included in Table\u200531P{1H} NMR spectra. Deprotonation at phosphorus was in all cases confirmed by the disappearance of the characteristic signal splitting due to 1JPH.1\u2009c]: Conversion >98\u2009%. 31P{1H} NMR (C6D6): \u03b4=61.8\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221232.9\u2005ppm ; 1H\u2005NMR ([D8]THF): \u03b4=3.56 , 1.12 , 1.02 , 0.33\u2005ppm ; 13C{1H} NMR ([D8]THF): \u03b4=47.3 , 23.9\u2005ppm : \u03b4=\u22120.39\u2005ppm (s).Li[1\u2009c]: 31P{1H} NMR (C6D6): \u03b4=53.4\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221229.5\u2005ppm ; 1H\u2005NMR (C6D6): \u03b4=3.75 , 1.43 , 1.23 , 0.47\u2005ppm ; 13C{1H} NMR (C6D6): \u03b4=47.9 , 24.7 , 24.2\u2005ppm .K[1\u2009c]: Conversion 94\u2009%. 31P{1H} NMR (C6D6): \u03b4=68.5\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221230.4\u2005ppm ; 1H\u2005NMR ([D8]THF): \u03b4=6.97\u20136.84 , 4.22\u20134.14 , 3.61 , 1.13 , 1.04 , 0.76\u2005ppm ; 13C{1H} NMR ([D8]THF): \u03b4=120.7 , 110.7 , 68.6 , 67.3 , 47.3 , 24.1\u2005ppm .[K(dibenzo\u201018\u2010crown\u20106)][1\u2009e]: 31P{1H} NMR (C6D6): \u03b4=185.7\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221232.2\u2005ppm ; 1H\u2005NMR ([D8]THF): \u03b4=6.97 , 4.1 , 3.99 , 3.26 , 1.22 , 1.18\u2005ppm ; 13C{1H} NMR ([D8]THF): \u03b4=150.6 , 149.1 , 143.1 , 124.1 , 122.9 , 54.2 , 27.4 , 25.0\u201324.5\u2005ppm .Li[1\u2009e]: 31P{1H} NMR (C6D6): \u03b4=187.7\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221229.9\u2005ppm ; 1H\u2005NMR ([D8]THF): \u03b4=6.94 , 4.22\u20133.80 , 3.16 , 1.25\u20131.12\u2005ppm ; 13C{1H} NMR ([D8]THF): \u03b4=150.5 , 149.4 , 143.3 , 124.3 , 123.1 , 122.7 , 54.4 , 27.3 , 25.5 , 25.2 , 24.1 , 23.1\u2005ppm .K[1\u2009e]: 31P{1H} NMR (C6D6): \u03b4=197.3\u2005ppm ; 11B{1H} NMR (C6D6): \u03b4=\u221229.8\u2005ppm ; 1H\u2005NMR ([D8]THF): \u03b4=6.95 , 6.82 , 4.27\u20134.04 ,3.98 , 3.84 , 3.22 , 1.20 , 1.15 , 0.67\u2005ppm ; 13C{1H} NMR ([D8]THF): \u03b4=142.2 , 122.1 , 118.8 , 108.8 , 66.6 , 65.3 , 52.6 , 25.6 , 22.9 , 22.7\u2005ppm .[K(dibenzo\u201018\u2010crown\u20106)] was prepared as described above from 2\u2009a , KHMDS and Et2O (15\u2005mL). CS2 was added through a microliter syringe. The solution was stirred for 18\u2005h. The solvent was evaporated under reduced pressure. The remaining work\u2010up was carried out in air. The residue extracted with Et2O (10\u2005mL). The extract was filtered in air over Celite, and pentane (10\u2005mL) was added. Storing the solution at \u221224\u2009\u00b0C afforded 8 as red crystalline solid . 31P{1H} NMR (C6D6): \u03b4=93.4\u2005ppm (broad); 11B{1H} NMR \u03b4=\u221234.0\u2005ppm (broad); 1H\u2005NMR (C6D6): \u03b4=3.35 , 3.20 , 1.19 , 1.07\u2005ppm ; 13C{1H} NMR (C6D6): \u03b4=42.0 (broad), 14.4 , detection of the CS2\u2010carbon atom was precluded by low S/N; C9H22BKN2PS2\u22c50.5 H2O (313.3\u2005g\u2009mol\u22121): calcd C 34.50 H 7.72 N 8.94 S 20.47; found C 33.85 H 7.49 N 8.67 S 19.93. The deviation between calculated and found analytical data is presumably due to a higher water content of the hygroscopic sample ; MS ((\u2212)\u2010ESI): m/z=264.11 [(M\u2212K)\u2212].The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "The title compound exists in the crystal as a dimer of ion pairs. Hydrogen bonding and weak \u03c0\u2013\u03c0 inter\u00adactions along with N\u2014H\u22ef\u03c0 inter\u00adactions are involved in consolidating this cluster. The three-dimensional crystal structure consists of stepped stacks of dimers of ion pairs associated by C\u2014H\u22ef\u03c0(ring) and slipped \u03c0-stacking inter\u00adactions. 21H15N2+\u00b7C7H5O2\u2212, 2-phenyl-1H-phenanthroimidazole and benzoic acid form an ion pair complex. The system is consolidated by hydrogen bonds along with \u03c0\u2013\u03c0 inter\u00adactions and N\u2014H\u22ef\u03c0 inter\u00adactions between the constituent units. For a better understanding of the crystal structure and inter\u00admolecular inter\u00adactions, a Hirshfeld surface analysis was performed.In the title compound, C The space group is monoclinic, P21/n and two asymmetric units, two M1+ ions and two benzoate ions, are combined in an inversion dimer of ion pairs \u00b0] to one another and the torsional angle C1\u2014O1\u2014N1\u2014C22 is 78.24 (su?)\u00b0. Unit A is stabilized by hydrogen bonds \u2005\u00c5].The structure of the title compound is shown in Fig.\u00a01A units are associated through weak, slipped, \u03c0-stacking inter\u00adactions between the C9\u2013C14 benzene rings and N1/C22/N2/C21/C8 imidazole rings across inversion centers .Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020005344/mw2157sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020005344/mw2157Isup2.hklStructure factors: contains datablock(s) I. DOI: 1997348CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "C(6) chains propgagating along [001].The title compound consists of two fluoro\u00adphenyl groups and one butyl group equatorially oriented on a piperidine ring, which adopts a chair conformation. The dihedral angle between the mean planes of the phenyl rings is 72.1\u2005(1)\u00b0. In the crystal, weak N\u2014H\u22efO and C\u2014H\u22efF inter\u00adactions, which form 21H23F2NO, consists of two fluoro\u00adphenyl groups and one butyl group equatorially oriented on a piperidine ring, which adopts a chair conformation. The dihedral angle between the mean planes of the phenyl rings is 72.1\u2005(1)\u00b0. In the crystal, N\u2014H\u22efO and weak C\u2014H\u22efF inter\u00adactions, which form R22[14] motifs, link the mol\u00adecules into infinite C(6) chains propagating along [001]. A weak C\u2014H\u22ef\u03c0 inter\u00adaction is also observed. A Hirshfeld surface analysis of the crystal structure indicates that the most significant contributions to the crystal packing are from H\u22efH (53.3%), H\u22efC/C\u22efH (19.1%), H\u22efF/F\u22efH (15.7%) and H\u22efO/O\u22efH (7.7%) contacts. Density functional theory geometry-optimized calculations were compared to the experimentally determined structure in the solid state and used to determine the HOMO\u2013LUMO energy gap and compare it to the UV\u2013vis experimental spectrum.The title compound, C Ramachandran et al., 2007et al., 2020Piperidin-4-one compounds have various biological properties and have applications as anti-viral, antitumor, and antihistaminic agents As part of our studies in this area, we now describe the synthesis and structure of the title compound, C1/cP2 with one mol\u00adecule in the asymmetric unit \u2005\u00c5, \u03b8 = 6.56\u2005(15)\u00b0, \u03c6 = 356.9\u2005(14)\u00b0. The dihedral angles for the C1\u2013C5/N1 piperidine (A), C6\u2013C11 fluorophenyl (B) and C12\u2013C17 fluorophenyl (C) rings are A/B = 65.50\u2005(8), A/C = 73.87\u2005(8) and B/C = 72.11\u2005(8)\u00b0. The substituents on the piperidine ring adopt equatorial orientations with the keto oxygen atom being anti-clinal [O1\u2014C3\u2014C2\u2014C1 = \u2212124.44\u2005(16)\u00b0]. The butyl group lies in a syn-periplanar orientation [O1\u2014C3\u2014C2\u2014C18 = 0.7\u2005(2)] while the fluoro\u00adphenyl groups are both anti-clinal [N1\u2014C5\u2014C6\u2014C7 = \u2212148.28\u2005(13) and N1\u2014C1\u2014C12\u2014C17 = \u221275.42\u2005(16)\u00b0]. The sum of the bond angles around N1 is 336.8\u00b0, which is consistent with sp3 hybridization for this atom it Fig.\u00a01. In the C(6) chains (via the N\u2014H\u22efO bond) along [001]. Some longer C\u2014H\u22efO and C\u2014H\u22efF contacts are also present as well as a single weak C\u2014H\u22ef\u03c0 inter\u00adaction n Table\u00a01.CrystalExplorer17.5 . The curvedness of the HS can be used to divide the mol\u00adecular surface into contact patches with each neighbouring mol\u00adecule thereby using it to define a coordination number in the crystal .A Hirshfeld surface (HS) analysis are the most important inter\u00adactions , presumably because of the large hydrogen content of (I)de + di) < 1.19\u2005\u00c5. The pair of wings for the H\u22efC/C\u22efH contacts is in the region 1.04\u2005\u00c5 < (de + di) < 1.58\u2005\u00c5 and includes the weak C\u2014H\u22ef\u03c0 inter\u00adaction. The H\u22efF/F\u22efH contacts are seen as a pair of wings in the region 1.04\u2005\u00c5 < (de + di) < 1.38\u2005\u00c5. The wings for the H\u22efO/O\u22efH contacts are in the region of 0.88\u2005\u00c5 < (de + di) < 1.20\u2005\u00c5 while the blunt wings in the plot for F\u22efF contacts are in the region 1.60\u2005\u00c5 < (de + di) < 1.70\u2005\u00c5. The C\u22efC contacts make a negligible 0.1% contribution and are viewed as a dash pattern pointing diagonally left. The O\u22efO contacts make no contribution to the HS. The most significant of these contributions to the overall Hirshfeld surface are shown in Figure S3 in the supporting information.Two-dimensional fingerprint plots show the relative contributions of the various types of contacts to the Hirshfeld surface for (I)ns Fig.\u00a04b, presuts Fig.\u00a04f; 2.6% ts Fig.\u00a04g make ats Fig.\u00a04h make nWebMo Pro \u00b0 with respect to each other.A density functional theory (DFT) geometry-optimized calculation for (I)supplementary Table S2. Both the HOMO and HOMO\u22121 are localized largely on the piperidine ring. For the LUMO, LUMO+1 and LUMO+2, the orbitals are delocalized over the piperidine ring as well as both phenyl rings. The observed UV/vis absorption spectrum . The molar extinction coefficients, \u220a, are 1.12 and 2.50\u2005l\u2005mol\u22121 cm\u22121, respectively. We tentatively assign the first absorption band envelope at 256\u2005nm to overlapping contributions from HOMO \u2192 LUMO (energy gap 5.71\u2005eV), HOMO \u2192 LUMO+1 (5.83\u2005eV) and HOMO\u22121 \u2192 LUMO (5.82\u2005eV). The band at 216\u2005nm is assigned to overlapping contributions from HOMO \u2192 LUMO+2 (5.89\u2005eV), HOMO\u22121 \u2192 LUMO+1 (5.95\u2005eV) and HOMO\u22121 \u2192 LUMO+2 (6.01\u2005eV).The calculated energies (eV) for the frontier mol\u00adecular orbitals are shown in Fig.\u00a05um Fig.\u00a06 shows twet al.. 2016et al., 2001t(3)-pentyl-r(2),c6)-di\u00adphenyl\u00adpiperidin-4-one ,c(6)-di\u00adphenyl\u00adpiperidin-4-one ,c(6)-bis\u00ad(4-fluoro\u00adphen\u00adyl)piperidin-4-one : found C 74.24, H 6.16, N 4.03; calculated C 73.45, H 6.75, N 4.08; melting point 381.5\u2005K.A mixture of ammonium acetate , 4-fluoro\u00adbenzaldehyde and 2-hepta\u00adnone in distilled ethanol was heated first to boiling. After cooling, the viscous liquid obtained was dissolved in ether (200\u2005ml) and shaken with 100\u2005ml concentrated hydro\u00adchloric acid. The precipitated hydro\u00adchloride of 3-butyl-2,6-bis\u00ad(4-fluoro\u00adphen\u00adyl)piperidin-4-one was removed by filtration and washed first with a 50\u2005ml mixture of ethanol and ether (1:1) and then with ether to remove most of the coloured impurities. The resulting yellowish base was liberated from an alcoholic solution by adding aqueous ammonia (15\u2005ml) and then diluted with water (200\u2005ml). Then, 1.0\u2005g of the crude sample was dissolved in 100\u2005ml of absolute alcohol, warmed until the sample dissolved, and 2.0\u2005g of animal charcoal added in the resulting solution. The hot solution was filtered and the procedure repeated again. The filtered solution was left for 48\u2005h and colourless prisms of (I)\u22121) (KBr): 3287 (\u03bdN\u2014H), 3134, 2929, 2866 (\u03bdC\u2014H), 1702 (\u03bdC=O), 1605, 1508 (\u03bdC=C), 793 (\u03bdC\u2014Cl); 1H NMR : \u03b4 7.01\u20137.45 , 4.04 , 3.68 , 2.67 , 2.56 , 2.0 (NH proton), 0.95\u20131.0 CH2(3), 1.09\u20131.15 CH2(2), 1.59\u20131.63 CH2(1), 0.74, ; 13C NMR : \u03b4 129.16, 129.38, 128.18, 128.10, 115.64, 115.56, 115.43, 115.35 (aromatic carbon atoms), 138.52 and 137.64 (aromatic ipso carbon atoms), 66.33 (C2), 57.50 (C3), 208.7 (C4), 51.63 (C5), 61.08 (C6), 24.30 C18H2, 29.71 C19H2, 22.75 C20H2, 13.81 C21H3.FT\u2013IR (cmUiso(H) = 1.2Ueq(carrier) or 1.5Ueq(methyl carrier) was applied in all cases.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020004636/hb7882sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989020004636/hb7882Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020004636/hb7882sup3.docxTheoretical chemistry data and Hirshfeld figures. DOI: 1994539CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Hirshfeld surface and simultaneous TGA\u2013DTA analyses are also describedThe mol\u00adecular and crystal structure of bis\u00ad, crystallizes with one half mol\u00adecule in the asymmetric unit of the monoclinic unit cell. The complex adopts an octa\u00adhedral coordination geometry with two mutually trans benzyl-2-(heptan-4-yl\u00adidene)hydrazine-1-carboxyl\u00adate ligands in the equatorial plane with the axial positions occupied by N-bound thio\u00adcyanato ligands. The overall conformation of the mol\u00adecule is also affected by two, inversion-related, intra\u00admolecular C\u2014H\u22efO hydrogen bonds. The crystal structure features N\u2014H\u22efS, C\u2014H\u22efS and C\u2014H\u22efN hydrogen bonds together with C\u2014H\u22ef\u03c0 contacts that stack the complexes along the b-axis direction. The packing was further explored by Hirshfeld surface analysis. The thermal properties of the complex were also investigated by simultaneous TGA\u2013DTA analyses.The title centrosymmetric Ni The coordination chemistry of benzyl carbazate Schiff base complexes has gained importance not only from the inorganic point of view, but also because of their biological and thermal properties. In the course of our recent studies on such complexes, we reported the cobalt(II) complex of a Schiff base derived from benzyl carbazate and heptan-4-one with thio\u00adcyanates as the charge-compensating ligands \u00b0. Bond lengths and angles in the closely related Ni and Co complexes are generally similar, although the Ni1\u2014N1 bond [2.1332\u2005(12)\u2005\u00c5] is significantly shorter here than the corresponding Co1\u2014N11 and Co2\u2014N21 vectors [2.206\u2005(5) and 2.248\u2005(6)\u2005\u00c5 respectively].The title compound, N\u22efS1 and weaker C8\u2014H8\u22efS1 and C10\u2014H10A\u22efS1 hydrogen bonds, Table\u00a01bc diagonal, Fig.\u00a02B\u22efS1 hydrogen bonds link adjacent mol\u00adecules into rows along the b-axis direction, Fig.\u00a03a, through C2\u2014H2A\u22efCg3, C\u2014H\u22ef\u03c0 contacts, Fig.\u00a04Cg3 is the centroid of the C3\u2013C8 phenyl ring. These contacts combine to stack mol\u00adecules of the complex in a regular fashion along the b-axis direction, Fig.\u00a05In the crystal structure, atom S1 acts as a trifurcated acceptor forming N2\u2014H21 were obtained using Hirshfeld surface analysis and (b). Bold red circles on the Hirshfeld surfaces correspond to the N\u2014H\u22efS hydrogen bonds while the weaker C\u2014H\u22efS and C\u2014H\u22ef\u03c0 contacts appear as faint red circles. Fingerprint plots, Fig.\u00a07Further details of the inter\u00admolecular inter\u00adactions in 1. Simultaneous TGA\u2013DTA analyses were recorded in air on a Perkin\u2013Elmer SII Thermal Analyser over the temperature range 50\u2013800\u00b0C. With the equipment used here, the TGA curve shows the temperature range but not the individual peak temperatures. However, peak temperatures can be seen in the DTA curve. In the first step of decomposition, the weight loss of 74% occurs over the temperature range 115\u2013260\u00b0C (TGA). This corresponds to the loss of the Schiff base ligands to form NiII thio\u00adcyanate as an inter\u00admediate. This was marked by both endothermic (170\u00b0C) and exothermic peaks (190 and 210\u00b0C) in the DTA curve. As the thermal analysis was carried out under a dynamic flowing air atmosphere, the S and N atoms are oxidized to SO2 and NO2, while nickel ultimately forms nickel oxide. Similar decomposition processes have been observed in our recent wok on numerous similar complexes, see for example NHN=C(CH2)2 unit produced only two hits. One was our own report of the ligand benzyl 2-cyclo\u00adpentyl\u00adidenehydrazine\u00adcarboxyl\u00adate -1-ethyl 8-methyl 7-(2-(benzyl\u00adoxycarbon\u00adyl)hydrazono)oct-2-enedioate, 2\u00b76H2O, dissolved in 10\u2005mL of doubly distilled water was added to this solution. The resulting blue solution was layered with heptan-4-one (dipropyl ketone) and the solution changed to a green colour. The final solution was left to evaporate at room temperature. After slow evaporation, bluish\u2013green rhombus-shaped crystals suitable for X-ray diffraction analysis were collected, washed with doubly distilled water and air-dried.Equimolar amounts of ammonium thio\u00adcyanate and benzyl carbazate were dissolved in methanol (10\u2005mL). Nickel nitrate, Ni and 3Ag2(F) \u21923Tg1(P) transitions, respectively, supporting the six-coordinate octa\u00adhedral geometry around the NiII cation spectrometer using tetra\u00admethyl\u00adsilane as an inter\u00adnal reference. Chemical shifts are expressed in parts per million (ppm): 0.84\u20130.88 and 1.33\u20132.20 ppm: CH3 and CH2 groups, respectively; \u2013OCH2 proton: 5.08 ppm; aromatic protons multiplets 7.29\u20137.34 ppm; NH: 9.882 ppm.The Simultaneous TGA\u2013DTA analyses were recorded in air on a PerkinElmer SII Thermal Analyser over the temperature range 50-800\u00b0C.Uiso(H) = 1.2Ueq(N). All C-bound H atoms were refined using a riding model with d(C\u2014H) = 0.95\u2005\u00c5, Uiso = 1.2Ueq(C) for aromatic 0.99\u2005\u00c5, Uiso = 1.2Ueq(C) for CH2 and 0.98\u2005\u00c5, Uiso = 1.5Ueq(C) for CH3 H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989020004260/vm2230sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020004260/vm2230Isup2.hklStructure factors: contains datablock(s) I. DOI: 1993291CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the complex mol\u00adecule, the tetra\u00addentate ligand N1,N3-bis\u00ad[methyl\u00adene]-2,2-di\u00admethyl\u00adpropane-1,3-di\u00adamine coordinates to the FeII ion through the N atoms of the 1,2,3-triazole moieties and aldimine groups. Two thio\u00adcyanate anions, coordinating through their N atoms, complete the coordination sphere of the central ion. In the crystal, neighbouring mol\u00adecules are linked through weak C\u2014H\u22ef\u03c0, C\u2014H\u22efS and C\u2014H\u22efN inter\u00adactions into a two-dimensional network extending parallel to (011). The inter\u00admolecular contacts were qu\u00adanti\u00adfied using Hirshfeld surface analysis and two-dimensional fingerprint plots, revealing the relative contributions of the contacts to the crystal packing to be H\u22efH (35.2%), H\u22efC/C\u22efH (26.4%), H\u22efS/S\u22efH (19.3%) and H\u22efN/N\u22efH (13.9%).The unit cell of the title compound, [Fe The average trigonal distortion parameters \u03a3 = \u03a3112(|90\u00a0\u2212\u00a0\u03c6i|), where \u03c6i is the angle N\u2014Fe\u2014N\u2032 , where \u03b8i is the angle generated by superposition of two opposite faces of an octa\u00adhedron . The volume of the [FeN6] coordination polyhedron is 12.4\u2005\u00c53.The Fens Fig.\u00a01. The aveB\u22efCgi of 3.330\u2005(3)\u2005\u00c5 \u2005\u00c5] with a thio\u00adcyanate anion. This, together with the C4\u2014H4B\u22efC27ii and C4\u2014H4B\u22efN10ii inter\u00adactions [3.709\u2005(3) and 3.617\u2005(3)\u2005\u00c5] involving the C\u2261N group of the anion, links the dimers into a supra\u00admolecular chain propagating parallel to [01iii = 3.603\u2005(3)\u2005\u00c5] and triazole groups [C19\u2014H19A\u22efN7iii = 3.311\u2005(3)\u2005\u00c5] of neighbouring complex mol\u00adecules, forming a two-dimensional supra\u00admolecular array extending parallel to (011).Neighbouring complex mol\u00adecules form dimers through double weak contacts C18\u2014H181] Fig.\u00a02. These cCrystal Explorer to 1.8236 (blue) a.u. The pale-red spots symbolize short contacts and negative dnorm values on the surface correspond to the inter\u00adactions described above. The overall two-dimensional fingerprint plot is illus\u00adtrated in Fig.\u00a03dnorm are shown for the H\u22efH, H\u22efC/C\u22efH, H\u22efS/S\u22efH, and H\u22efN/N\u22efH contacts, and the two-dimensional fingerprint plots are presented in Fig.\u00a04Hirshfeld surface analysis was performed and the associated two-dimensional fingerprint plots were generated using et al., 2016II thio\u00adcyanate complexes, derivatives of a 1,3-di\u00adamino\u00adpropanes and N-substituted 1,2,3-triazole aldehydes, viz. DURXEV, ADAQUU, ADAREF and solvatomorphs ADAROP and ADARUV 4(NCS)2] dissolved in a minimum amount of boiling methanol with a minimum amount of ascorbic acid. The formed yellow solution was slowly cooled to ambient temperature. The formed orange crystals were subsequently filtered off. Elemental analysis calculated (%) for C27H28FeN10S2: C, 52.94; H, 4.61; N, 22.87; S, 10.47; found: C, 52.88; H, 4.37; N, 22.40; S, 10.35. IR vKBr (cm\u22121): 1615 (C=N), 2071, 2115 (NCS).The ligand of the title compound was obtained Uiso(H) = 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989020012608/wm5580sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020012608/wm5580Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020012608/wm5580Isup3.cdxSupporting information file. DOI: 2032292CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, centrosymmetric, eight-membered {\u22efHNCS}2 synthons are formed by thio\u00adamide-N\u2014H\u22efS(thio\u00adamide) hydrogen bonds.The title hydrazine carbodi\u00adthio\u00adate features an almost planar C 13H18N2OS2, is constructed about a central and almost planar C2N2S2 chromophore (r.m.s. deviation = 0.0263\u2005\u00c5); the terminal meth\u00adoxy\u00adbenzene group is close to coplanar with this plane [dihedral angle = 3.92\u2005(11)\u00b0]. The n-butyl group has an extended all-trans conformation . The most prominent feature of the mol\u00adecular packing is the formation of centrosymmetric eight-membered {\u22efHNCS}2 synthons, as a result of thio\u00adamide-N\u2014H\u22efS(thio\u00adamide) hydrogen bonds; these are linked via meth\u00adoxy-C\u2013H\u22ef\u03c0(meth\u00adoxy\u00adbenzene) inter\u00adactions to form a linear supra\u00admolecular chain propagating along the a-axis direction. An analysis of the calculated Hirshfeld surfaces and two-dimensional fingerprint plots point to the significance of H\u22efH (58.4%), S\u22efH/H\u22efS (17.1%), C\u22efH/H\u22efC (8.2%) and O\u22efH/H\u22efO (4.9%) contacts in the packing. The energies of the most significant inter\u00adactions, i.e. the N\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions have their most significant contributions from electrostatic and dispersive components, respectively. The energies of two other identified close contacts at close to van der Waals distances, i.e. a thione\u2013sulfur and meth\u00adoxy\u00adbenzene\u2013hydrogen contact and between methyl\u00adene-H atoms , are largely dispersive in nature.The title hydrazine carbodi\u00adthio\u00adate, C The meth\u00adoxy\u00adbenzene ring forms a dihedral angle of 3.92\u2005(11)\u00b0 with the central residue indicating a close to co-planar relationship. The C9\u2014O1\u2014C6 \u2014C7 dihedral angle of 176.9\u2005(3)\u00b0 indicates that the meth\u00adoxy substituent lies almost in the plane of the benzene ring to which it is connected. The configuration about the C2=N2 imine bond [1.278\u2005(3)\u2005\u00c5] is E and this bond length is significantly shorter than the C1\u2014N1 bond [1.330\u2005(3)\u2005\u00c5]; the N1\u2014N2 bond length is 1.378\u2005(3)\u2005\u00c5. There is a large disparity in the C1\u2014S1 [1.662\u2005(3)\u2005\u00c5] and C1\u2014S2 [1.745\u2005(3)\u2005\u00c5] bond lengths, which correlate with significant double-bond character in the former; the C10\u2014S2 bond length at 1.793\u2005(3)\u2005\u00c5 is longer than each of these. The thione character of the C1\u2014S1 bond is also reflected in the range of angles subtended at the C1 atom, which are systematically wider for those involving the thione-S1 atom, i.e. S1\u2014C1\u2014S2 [126.35\u2005(16)\u00b0] and S1\u2014C1\u2014N1 [120.9\u2005(2)\u00b0], cf. S2\u2014C1\u2014N1 [112.76\u2005(19)\u00b0]. The thio\u00adamide-N\u2014H and thio\u00adamide-S atoms have a syn disposition. Finally, the n-butyl group has an extended, all-trans conformation as seen in the S2\u2014C10\u2014C11\u2014C12 [\u2212173.2\u2005(3)\u00b0] and C10\u2014C11\u2014C12\u2014C13 [180.0\u2005(4)\u00b0] torsion angles.The mol\u00adecular structure of (I)via an eight-membered {\u22efHNCS}2 synthon, the mol\u00adecular packing is largely devoid of directional inter\u00adactions inter\u00adactions, Fig.\u00a02a), being the only other identified supra\u00admolecular association. Globally, chains pack without specific inter\u00adactions between them, Fig.\u00a02b). An analysis of the weak non-covalent contacts within and connecting chains is given in the Analysis of the Hirshfeld surfaces.With the exception of thio\u00adamide-N\u2014H\u22efS(thio\u00adamide) hydrogen bonding between centrosymmetrically related mol\u00adecules, Table\u00a01et al., 2019Crystal Explorer 17 a)\u2013(e), respectively; the percentage contributions from the different inter\u00adatomic contacts are summarized in Table\u00a02b), a short inter\u00adatomic H\u22efH contact involving methyl\u00adene-H10B with a symmetry-related mate and occurring between supra\u00admolecular chains aligned along the a axis, is observed as a single peak at de + di \u223c 2.2\u2005\u00c5. In the fingerprint delineated into S\u22efH/H\u22efS contacts, shown in Fig.\u00a06c), the pair of well-defined spikes at de + di \u223c 2.5\u2005\u00c5 arise as a result of the prominent inter\u00admolecular N\u2014H\u22efS inter\u00adaction. The points corresponding to S\u22efH/H\u22efS contacts involving the thione-S1 and meth\u00adoxy\u00adbenzene-H4 atoms, occurring within the supra\u00admolecular chain shown in Fig.\u00a02a), albeit at nearly van der Waals separations , and reflected as an electrostatic inter\u00adaction in the Hirshfeld surface plotted over the electrostatic potential of Fig.\u00a04d) are at distances equal to or greater than the sum of van der Waals radii, the presence of characteristic wings is the result of the inter\u00admolecular methy\u00adoxy-C\u2014H\u22ef\u03c0(meth\u00adoxy\u00adbenzene) contact. The points corresponding to inter\u00adatomic O\u22efH/H\u22efO contacts illustrated in the corres\u00adponding fingerprint plot of Fig.\u00a06e), also show a pair of forceps-like tips at de + di \u223c 2.8\u2005\u00c5, i.e. at van der Waals distances. The contribution from the other inter\u00adatomic contacts summarized in Table\u00a02The overall two-dimensional fingerprint plot for (I)Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) level of theory. The nature and strength of the inter\u00admolecular inter\u00adactions in terms of their energies are qu\u00adanti\u00adtatively summarized in Table\u00a03Eele, Edisp and Etot components, respectively; the radius of the cylinder is proportional to the magnitude of the inter\u00adaction energies. This is reflected in the relatively thick red cylinders corresponding to the electrostatic inter\u00adactions via the N\u2014H\u22efS hydrogen bonding in Fig.\u00a07a) and the thick green cylinders corresponding to the strong dispersive inter\u00adactions provided by the methy\u00adoxy-C\u2014H\u22ef\u03c0(meth\u00adoxy\u00adbenzene) inter\u00adactions in Fig.\u00a07b).The pairwise inter\u00adaction energies between mol\u00adecules in the crystal of (I)6H4C(H)=NN(H)C(=S)SR. These are of the R = Me Reflecting the inter\u00adest in the chemistry of hydrazine carbodi\u00adthio\u00adates related to (I)S-butyl\u00addithio\u00adcarbazate (SBuDTC). An ethano\u00adlic solution (28\u2005ml) of 4-meth\u00adoxy\u00adbenzaldehyde was added directly to the SBuDTC in situ. This mixture was heated to 323\u2005K with continuous stirring for 30\u2005min. The yellow product (I)13H18N2OS2: C, 55.3; H, 6.4; N, 9.9; S, 22.7. Found: C, 55.9; H, 6.6; N, 9.8; S, 23.2. FT\u2013IR (cm\u22121): 3120 \u03bd(NH), 2927 \u03bd(CH), 1600 \u03bd(C=N), 1248 and 1107 \u03bd(COC), 1017 \u03bd(NN), 861 \u03bd(CSS). MS: calculated m/z = 282; Found m/z = 282. 1H NMR : \u03b4 13.11 , 8.15 , 6.97, 7.02, 7.61, 7.78 (ArH), 3.76 , 3.14 , 1.58 , 1.36 , 0.86 . 13C{1H} NMR : \u03b4 196.96 (C=S), 146.87 (C=N), 161.90, 129.65, 126.36, 115.00 (ArC), 55.86 (OCH3), 33.19, 31.05, 22.10 (CH2), 14.07 (CH3). NMR data were measured on a JOEL ECX500 FT NMR spectrometer.In an ice-bath, carbon di\u00adsulfide was added dropwise to an absolute ethanol (35\u2005ml) solution comprising KOH and hydrazine hydrate . After 30\u2005min, 1-bromo\u00adbutane was added. The solution was stirred at 278\u2005K for 1\u2005h to form Uiso(H) set to 1.2Ueq(C). The N-bound H atom was located in a difference-Fourier map but was refined with a N\u2014H distance restraint of 0.86\u2005(1)\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989020000328/hb7887sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020000328/hb7887Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020000328/hb7887Isup3.cmlSupporting information file. DOI: 1977066CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "V with the dihedral angle between the phthalazin-1-one and mesityl residues being 83.26\u2005(4)\u00b0. Mol\u00adecules assemble into a linear, supra\u00admolecular tape by phthalazinone-C6-C\u2014H\u22efO(sulfoxide) and \u03c0\u2013\u03c0 stacking inter\u00adactions.The mol\u00adecule in the title crystal has the shape of the letter 17H16N2O3S {systematic name: 2-[\u00adsulfon\u00adyl]-1,2-di\u00adhydro\u00adphthalazin-1-one}, features a tetra\u00adhedral sulfoxide-S atom, connected to phthalazin-1-one and mesityl residues. The dihedral angle [83.26\u2005(4)\u00b0] between the organic substituents is consistent with the mol\u00adecule having the shape of the letter V. In the crystal, phthalazinone-C6-C\u2014H\u22efO(sulfoxide) and \u03c0\u2013\u03c0 stacking [inter-centroid distance = 3.5474\u2005(9)\u2005\u00c5] contacts lead to a linear supra\u00admolecular tape along the a-axis direction; tapes assemble without directional inter\u00adactions between them. The analysis of the calculated Hirshfeld surfaces confirm the importance of the C\u2014H\u22efO and \u03c0-stacking inter\u00adactions but, also H\u22efH and C\u2014H\u22efC contacts. The calculation of the inter\u00adaction energies indicate the importance of dispersion terms with the greatest energies calculated for the C\u2014H\u22efO and \u03c0-stacking inter\u00adactions.The X-ray crystal structure of the title phthalazin-1-one derivative, C H)-one derivatives are a group of di\u00adaza\u00adheterobicycles that are noteworthy for their inter\u00adesting medicinal applications. Thus, this class of compound has been reported to possess a wide variety of biological properties such as anti-diabetic , anti-inflammatory and analgesic -one-based drugs are in use -ones, have also been reported -one was described. In the present study, the title compound, 2-[\u00adsulfon\u00adyl]-1,2-di\u00adhydro\u00adphthalazin-1-one, (I)Phthalazin-1\u00b0 for N1\u2014S1\u2014C1, involving the singly-bonded N1 and C1 atoms, to a wide 118.39\u2005(6)\u00b0, for O1\u2014S1\u2014O2, involving the doubly-bonded sulfoxide-O1, O2 atoms. The organic residues lie to the opposite side of the mol\u00adecule to the SO2 residue, forming dihedral angles of 67.35\u2005(4)\u00b0 [phthalazin-1-one with r.m.s. deviation = 0.0105\u2005\u00c5] and 49.79\u2005(6)\u00b0 [mesit\u00adyl]. The dihedral angle between the organic residues of 83.26\u2005(4)\u00b0 indicates a close to orthogonal relationship. The N2\u2014N1\u2014C10\u2014O3 torsion angle of \u2212179.88\u2005(12)\u00b0 indicates a co-planar arrangement for these atoms, which allows for the close approach of the N2 and O3 atoms, i.e. 2.6631\u2005(15)\u2005\u00c5, suggestive of a stabilizing contact \u2005\u00c5 and C10\u2014N1 = 1.4003\u2005(17)\u2005\u00c5. In each of the C17=N2 [1.2911\u2005(18)\u2005\u00c5] and C10=O3 [1.2175\u2005(15)\u2005\u00c5] bonds, double-bond character is noted. The bond angles about the N1 atom are non-symmetric, with the endocyclic N2\u2014N1\u2014C10 angle of 126.97\u2005(11)\u2005\u00c5 being significantly wider than the exocyclic N2\u2014N1\u2014S1 [113.93\u2005(9)\u2005\u00c5] and C10\u2014N1\u2014S1 [118.89\u2005(8)\u2005\u00c5] angles.The mol\u00adecule of (I)6-C\u2014H\u22efO(sulfoxide) contacts, Table\u00a01a). The \u03c0-stacking occurs between centrosymmetrically related phthalazinone rings, i.e. between the N2C4 and C6i rings with an inter-centroid distance = 3.5474\u2005(9)\u2005\u00c5, angle of inclination = 1.17\u2005(7)\u00b0 for symmetry operation (i) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 2\u00a0\u2212\u00a0z. As shown in Fig.\u00a02b), the tapes inter-digitate along the c-axis direction allowing for putative \u03c0-stacking between mesityl rings but, the inter-centroid separation is long at 4.1963\u2005(8)\u2005\u00c5. The assemblies shown in Fig.\u00a02b) stack along the a-axis direction, again without directional inter\u00adactions between them, Fig.\u00a02c).The formation of a supra\u00admolecular tape sustained by phthal\u00ada\u00adzinone-CCrystal Explorer 17 ,(b), the presence of diminutive red spots near methyl-C7 and benzene-H5 are indicative of inter\u00admolecular C\u2014H\u22efC contacts as C\u2014H\u22ef\u03c0 contacts are not preferred because of the V-shaped mol\u00adecular geometry of (I)6 ring on the dnorm-mapped Hirshfeld surface in Fig.\u00a03b) is indicative of short intra-chain C\u22efC contacts [Table\u00a02a)] and is consistent with the significant contribution from \u03c0\u2013\u03c0 stacking between centrosymmetrically related phthalazinone-N2C4 and C6 rings, encompassing connections between phthalazinone-C6 rings [3.6657\u2005(9)\u2005\u00c5 with angle of inclination = 0.03\u2005(7)\u00b0]. The involvement of the methyl-C8 atom in C\u2014H\u22efO [to provide links between the chains shown in Fig.\u00a02b)] and C\u2014H\u22efC contacts, Table\u00a02c). The blue and red regions corres\u00adponding to positive and negative electrostatic potentials, respectively, on the Hirshfeld surface mapped over electrostatic potential shown in Fig.\u00a04In order to probe the inter\u00adactions between mol\u00adecules of (I) al. 2019. In addia)\u2013(e), respectively; the percentage contributions from the different inter\u00adatomic contacts to the Hirshfeld surfaces are summarized in Table\u00a03A atoms, Table\u00a02de + di \u223c2.2\u2005\u00c5 in the fingerprint plot delineated into H\u22efH contacts, Fig.\u00a05b). In the fingerprint plot delineated into O\u22efH/H\u22efO contacts illustrated in Fig.\u00a05c), a pair of forceps-like tips at de + di \u223c2.3\u2005\u00c5, indicate the inter\u00admolecular C\u2014H\u22efO inter\u00adaction involving the phthalazinone-H12 and sulfoxide-O2 atoms, whereas the other inter\u00adatomic O\u22efH/H\u22efO contacts are merged within the plot and appear as a pair of intense blue spikes at de + di \u223c2.8\u2005\u00c5. Despite the observation that inter\u00admolecular C\u2014H\u22ef\u03c0 contacts are usually preferred by methyl groups, none are found involving those substituted at (C1\u2013C6) benzene ring in the crystal due to the V-shaped geometry. Rather, the involvement of methyl-C7 and H5A atoms, and benzene-C5 and H7C atoms [to provide links between the chains shown in Fig.\u00a02b)] in C\u2014H\u22efC inter\u00adactions, Table\u00a02de + di \u223c2.8\u2005\u00c5 in the fingerprint plot delineated into C\u22efH/H\u22efC contacts, Fig.\u00a05d). The presence of \u03c0\u2013\u03c0 stacking inter\u00adactions between symmetry-related phthalazinone-N2C4 and C6 rings is also evident as the arrow-shaped distribution of points around de, di \u223c1.8\u2005\u00c5 in the fingerprint plot delineated into C\u22efC contacts, Fig.\u00a05e). The contribution from other inter\u00adatomic contacts, summarized in Table\u00a02The overall two-dimensional fingerprint plots for (I)Eele), polarization (Epol), dispersion (Edis) and exchange\u2013repulsion (Erep) following Turner et al. level of theory. The nature and strength of the inter\u00admolecular inter\u00adactions in terms of their energies are qu\u00adanti\u00adtatively summarized in Table\u00a042C4 and C6 rings and the short inter\u00adatomic O1\u22efH14 contact have the greatest energy. The short inter\u00adatomic C5\u22efH7C, O3\u22efH8A and C10\u22efH8A contacts also have significant inter\u00adaction energies due to their participation in inversion-related contacts. Lower energies, compared to above inter\u00adactions, are calculated for the H12\u22efH9A, C7\u22efH5 and O1\u22efH9C contacts.The pairwise inter\u00adaction energies between the mol\u00adecules within the crystal of (I) al. 2017. The eneEele, Edisp and Etot, respectively. The images emphasize the importance of dispersion inter\u00adactions in the mol\u00adecular packing.Fig.\u00a06H)-one et al., 20182CCH3)3\u00b74H2O was added with constant stirring to an ethanol solution (20\u2005ml) of (III) . The resulting mixture was refluxed for 3\u2005h in an oil bath. The obtained colourless solution was concentrated to afford a colourless precipitate, which was filtered, dried under suction and further dried in vacuo over CaCl2. The precipitates were dissolved in ethanol, the resultant colourless solution was filtered and left at room temperature for 48\u2005h to obtain colourless crystals of (I)2-{[2-hydrazinyl\u00adidene]meth\u00adyl}benzoic acid (III) was obtained by a method reported earlier (Asegbeloyin Uiso(H) set to 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989020005101/lh5956sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020005101/lh5956Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020005101/lh5956Isup3.cmlSupporting information file. DOI: 1996401CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound belongs to the class of neutral pyridine aroylhydrazone complexes. In the crystal, \u03c0\u2013\u03c0 stacking leads to dimerization of the complex mol\u00adecules, which, in turn, are connected by weak C\u22efO and C\u22efC inter\u00adactions, thus giving rise to a supra\u00admolecular layered architecture. 17H18N3O4)2]\u00b7CH4O\u00b7C2H6O, contains two complex mol\u00adecules related by an inversion centre, plus one methanol and one ethanol solvent molecule per complex molecule. In each complex, two deprotonated pyridine aroylhydrazone ligands {3,4,5-trimeth\u00adoxy-N\u2032-[1-(pyridin-2-yl)ethyl\u00adidene]benzohydrazide} coordinate to the ZnII ion through the N atoms of the pyridine group and the ketamine, and, additionally, through the O atom of the enolate group. In the crystal, dimers are formed by \u03c0\u2013\u03c0 inter\u00adactions between the planar ligand moieties, which are further connected by C\u22efO and C\u22efC inter\u00adactions. The inter\u00admolecular inter\u00adactions were investigated using Hirshfeld surface analysis and two-dimensional fingerprint plots, revealing that the most important contributions for the crystal packing are from H\u22efH (44.8%), H\u22efC/C\u22efH (22.2%), H\u22efO/O\u22efH (18.7%) and C\u22efC (3.9%) inter\u00adactions.The unit cell of the title compound, [Zn(C These dynamic reversible properties have led to the exploration of charged and neutral spin-crossover iron(II) and iron(III) complexes, some with multifunctional properties /24, where \u03b8i is the angle generated by superposition of two opposite faces of the octa\u00adhedron /12, where \u03c6i is the deviation from 90\u00b0 of the cis-N\u2014Zn\u2014N angles in the coordination sphere \u2005\u00c5. In the dimer, the Zn\u22efZni separation is 7.612\u2005(2)\u2005\u00c5 \u2212x, \u2212y, \u2212z\u00a0+\u00a01] = 3.100\u2005(5)\u2005\u00c5 (Table\u00a01ii inter\u00addimer separation of 6.965\u2005(5)\u2005\u00c5. It is worth noting that a related FeII pyridine-based complex with butyl substituents consisting of uniform supra\u00admolecular chains with Fe\u22efFe separation of 7.676\u2005\u00c5 has previously been described \u2005\u00c5 and b = 13.8056\u2005(8)\u2005\u00c5. There are inter\u00adchain contacts C33\u22efC34iii/C34\u22efC33iii , below the sum of the van der Waals radii, between the meth\u00adoxy groups of neighbouring supra\u00admolecular chains at 3.385\u2005(5)\u2005\u00c5.The ligand mol\u00adecules exhibit slipped parallel \u03c0\u2013\u03c0 stacking between coplanar ligands of neighbouring mol\u00adecules, thus forming a dimeric structure; the closest C4\u22efC61] Fig.\u00a02. Neighbo\u00c5 Table\u00a01, with thiv between the ethanol methyl group and a meth\u00adoxy methyl group is 3.300\u2005(5)\u00c5. Additionally, neighbouring mol\u00adecules of ethanol are mutually bound forming dimers with C36\u22efC37v and O10\u22efC37v contacts with distances of 3.227\u2005(5) and 2.751\u2005(2)\u2005\u00c5, respectively. Furthermore, the co-crystallized mol\u00adecules of methanol form O\u2014H\u22efO hydrogen bonds with the meth\u00adoxy group of the ligand, with an O9\u22efO2 separation between the O atoms of 2.776\u2005(4)\u2005\u00c5.The neutral nature of the complex mol\u00adecule and therefore the absence of anions and, on the other hand, the relatively large size of the planar rigid substituents prevent the formation of a tightly packed lattice. Therefore, inter\u00admolecular voids are filled by the co-crystallized mol\u00adecules of ethanol, which act as bridges connecting the closest complex mol\u00adecules by O\u2014H\u22efN hydrogen bonding, with the distance between the donor and acceptor atoms O10\u22efN6 equal to 2.825\u2005(5)\u2005\u00c5. The contact C15\u22efC37CrystalExplorer17.5 software to 2.2951 (blue) a.u. The pale-red spots symbolize short contacts and negative dnorm values on the surface correspond to the inter\u00adactions described above. The overall two-dimensional fingerprint plot is illustrated in Fig.\u00a03dnorm are shown for the H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH, and C\u22efC contacts, and the two-dimensional fingerprint plots are presented in Fig.\u00a04The Hirshfeld surface analysis and the associated two-dimensional fingerprint plots were undertaken using et al., 2016N\u2032-[1-(pyridin-2-yl)ethyl\u00adidene]benzohydrazide ethyl\u00adidene]benzohydrazide ethyl\u00adidene]benzohydrazide ethyl\u00adidene]benzohydrazide to 177.76\u00b0 (MAKLES), while inter\u00admediate values of 168.09 and 170.56\u00b0 are observed for PATXAK and HIGPOD, respectively.A search of the Cambridge Structural Database to give a colourless complex. A pale-yellow solution was obtained after deprotonation with NEt3 (1\u2005mmol). The neutral complex was isolated by slow cooling the solution to ambient temperature and subsequently by filtering off the yellowish crystals. Elemental analysis calculated (%) for C37H46N6O10Zn: C 55.54, H 5.79, N 10.50; found: C 55.86, H 5.31, N 10.84. IR \u03bdKBr (cm\u22121): 1617 (N=C\u2014O), 1588, 1461 (C=Npy + C=CAr), 1252 (C\u2014O). MS ESI m/z (relative intensity): theoretically calculated 721.19 [M + H+] (100.0%). Found 721.21 [M + H+] (100.0%). TGA (up to 400\u2005K) expected weight loss for EtOH + MeOH: 9.8%; found: 9.5%.The complex was obtained by condensation of 3,4,5-tri\u00admeth\u00adoxy\u00adbenzohydrazide (1\u2005mmol) and acetyl pyridine (1.1\u2005mmol) in a mixture of absolute MeOH and EtOH (1:1) overnight in the presence of two drops of glacial acetic acid. The ligand obtained Uiso(H) = 1.2\u20131.5Ueq(C). None of the hydrogen atoms of the methanol or ethanol molecules could be located.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020000857/tx2017sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020000857/tx2017Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020000857/tx2017Isup3.cdxSupporting information file. DOI: 1979477CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(6) ring motif.The title mol\u00adecule is planar with an r.m.s. deviation for all non-hydrogen atoms of 0.018\u2005\u00c5. An intra\u00admolecular O1\u2014H1\u22efO5 hydrogen bond involving the adjacent hydroxyl and nitro groups closes an 8H7NO5, is planar with an r.m.s. deviation for all non-hydrogen atoms of 0.018\u2005\u00c5. An intra\u00admolecular O\u2014H\u22efO hydrogen bond involving the adjacent hy\u00addroxy and nitro groups forms an S(6) ring motif. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds, forming chains propagating along the b-axis direction. The chains are linked by C\u2014H\u22efO hydrogen bonds, forming layers parallel to the bc plane. The layers are linked by a further C\u2014H\u22efO hydrogen bond, forming slabs, which are linked by C=O\u22ef\u03c0 inter\u00adactions, forming a three-dimensional supra\u00admolecular structure. Hirshfeld surface analysis was used to investigate inter\u00admolecular inter\u00adactions in the solid state. The mol\u00adecule was also characterized spectroscopically and its thermal stability investigated by differential scanning calorimetry and by thermogravimetric analysis.The title compound, C Several synthetic routes for the synthesis of entacapone have been reported , was synthesized and its spectroscopic and structural features were studied. There are two reasons for this study, one is connected with the utilization of crystal structures in the identification of materials in the solid state, and the other is to build a library of structurally related compounds of entacapone that will be utilized in a future crystallization study.The title compound is a key starting material in the preparation of entacapone ring motif. The mol\u00adecule is planar with the maximum deviation from the mean plane being 0.038\u2005(1)\u2005\u00c5 for atom O5. The bonds lengths and bond angles are close to those found for similar structures (see \u00a74. Database survey).The mol\u00adecular structure of the title compound I, mol\u00adecules are linked by inter\u00admolecular bifurcated hydrogen bonds involving the hydroxyl group. Details of the hydrogen bonding together with the symmetry codes are given in Table\u00a01ialdehyde hydrogen bonds [2.6989\u2005(12)\u2005\u00c5] link the mol\u00adecules into chains with a C(8) motif. Each oxygen atom is involved in one or more inter\u00admolecular hydrogen bonds, except for the O5nitro atom, which is involved only in the intra\u00admolecular hydrogen bond. Each mol\u00adecule is connected by six adjacent mol\u00adecules through strong O1\u2014H1\u22efO3aldehyde and weak C7\u2014H7\u22efO4iinitro and C8\u2014H8A\u22efO3iiialdehyde hydrogen bonds, forming undulating layers parallel to the bc plane, enclosing two type of ring motifs \u2013 B\u22efO4ivnitro, forming slabs \u2005\u00c5] and C2\u2014O1\u22ef\u03c0 [3.353 (su?) \u00c5] close contacts Fig.\u00a02. The laybs Fig.\u00a03. Moreovets Fig.\u00a04 are preset al., 2016viz. a tin complex of the 4-hy\u00addroxy-3-meth\u00adoxy-5-nitro\u00adbenzaldehyde with a deprotonated hydroxyl group and benzyl anions -2-cyano-3--N,N-di\u00adethyl\u00adprop-2-enamide 1,3-di\u00admethyl-3,7-di\u00adhydro-1H-purine-2,6-dione monohydrate -2-cyano-3--N,N-di\u00adethyl\u00adprop-2-enamide pyridine-4-carboxamide -2-cyano-3--N,N-di\u00adethyl\u00adprop-2-enamide pyrazine-2-carboxamide -2-cyano-3--N,N-di\u00adethyl\u00adprop-2-enamide acetamide , while the most negative region is around the carbonyl oxygen atom . Those two atoms are involved in the shortest inter\u00admolecular hydrogen bond in the crystal structure (O1\u2014H1\u22efO3i), where O1\u22efO3i = 2.6989\u2005(12)\u2005\u00c5; see Table\u00a01Electrostatic potentials were calculated using ce Fig.\u00a07 in the ece Fig.\u00a07a, whileom Fig.\u00a07b. Those3 (10.5\u2005ml) was added dropwise over 30\u2005min. The reaction mixture was stirred for 30\u2005min at 283\u2013288\u2005K and 30\u2005min at 293\u2013298\u2005K. The suspension was then filtered and the crystals obtained were washed with water (3 \u00d7 20\u2005ml). The crystals were dried in a vacuum dryer to obtain pure yellow compound I . Yellow block-like crystals, suitable for X-ray diffraction analysis, were obtained by slow evaporation of a solution in acetone after 10\u2005d at room temperature.4-Hy\u00addroxy-3-meth\u00adoxy\u00adbenzaldehyde was dissolved in acetic acid (200\u2005ml) and the solution was cooled to 283\u2013288\u2005K and 65% HNOSpectroscopic analysis:I and 125.8\u2005MHz (13C) in CD3OD ; see Tables 2The structure of compound I Fig.\u00a08 was confI was investigated in the solid state by differential scanning calorimetry (DSC) and by thermogravimetric analysis (TGA). DSC analysis was performed on a TA Instruments Discovery DSC in a closed aluminium pan (40\u2005\u00b5L) under nitro\u00adgen flow (50\u2005ml\u2005min\u22121) and a heating rate of 10\u00b0C min\u22121 in the temperature range 25\u2013300 \u00b0C and a heating rate of 10\u00b0C min\u22121 in the temperature range 25\u2013300\u00b0C technique. It shows a broad band at about 3200\u2005cm\u22121, which corresponds to the O\u2014H stretching vibrations. Strong stretching vibrations of C=O and C\u2014O (aromatic ether) appear at 1683 and 1266\u2005cm\u22121, respectively. Bands corresponding to N\u2014O asymmetric and symmetric stretching modes can be found at 1547 and 1366\u2005cm\u22121, respectively. Characteristic weak overtones of the aromatic ring can be seen at 1800\u20131700\u2005cm\u22121.The IR spectrum Fig.\u00a012 of compoUiso(H) = 1.5Ueq(O) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989020000225/su5534sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989020000225/su5534Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020000225/su5534Isup3.cmlSupporting information file. DOI: 1957893CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I tricarbonyl complexes bearing ester-substituted bi\u00adpyridine or bi\u00adquinoline ligands is reported.The structural comparison of two Mn fac-bromido\u00adtricarbon\u00adylmangan\u00adese(I), [MnBr(C16H16N2O4)(CO)3], I, and fac-bromido\u00adtricarbon\u00adylmanganese(I), [MnBr(C24H20N2O4)(CO)3], II. In both complexes, the manganese(I) atom adopts a distorted octa\u00adhedral coordination sphere defined by three carbonyl C atoms, a Br\u2212 anion and two N atoms from the chelating \u03b1-di\u00adimine ligand. Both complexes show fac configurations of the carbonyl ligands. In I, the complex mol\u00adecules are linked by C\u2014H\u22efBr hydrogen bonds and aromatic \u03c0\u2013\u03c0 contacts. In II, intra\u00admolecular C\u2014H\u22efO hydrogen bonds are present as well as inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efBr hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions.The crystal structures of two manganese(I) complexes with ester-substituted bi\u00adpyridine or bi\u00adquinoline supporting ligands are reported, namely, We successfully compared their crystal structures and photophysical properties. As expected, a CT band shift in the visible region was confirmed, depending on the size of the \u03c0-conjugation system in \u03b1-di\u00adimine ligands. This finding will provide information in the future design of suitable complexes for a variety of photoreactions and 2.047\u2005(2)\u2005\u00c5, while in II, the Mn\u2014N bond lengths are 2.063\u2005(2) and 2.068\u2005(2)\u2005\u00c5. In I and II, the fac configuration of three CO ligands around the central manganese(I) atom is in agreement with their IR data. On the basis of their bond parameters, all CO ligands have typical triple-bond characters.The mol\u00adecular structures of compounds I (C3\u2014Mn1\u2014N1\u2014C8 and C2\u2014Mn1\u2014N2\u2014C9) are \u2212169.17\u2005(15) and 168.81\u2005(14)\u00b0, respectively; the corresponding torsion angles in II (C3\u2014Mn1\u2014N1\u2014C12 and C2\u2014Mn1\u2014N2\u2014C13) are \u2212147.52\u2005(16) and 147.08\u2005(17)\u00b0, respectively in debqn, and the equatorial CO ligands (C3\u2261O3 and C2\u2261O2). On the basis of similar steric hindrance, comparable torsion angles [150.4\u2005(15) and \u2212150.7\u2005(5)\u00b0] have been also observed in the related ReI complex \u2005\u00c5 and C10\u2014C11 = 1.392\u2005(3)\u2005\u00c5] are considerably longer than the corresponding one in II [C10\u2014C11 = 1.364\u2005(4)\u2005\u00c5 and C14\u2014C15 = 1.368\u2005(4)\u2005\u00c5]. This difference in structural parameters may eventually affect the intra\u00admolecular hydrogen-bond formation.Despite similar mol\u00adecular skeletons, only g Table\u00a02. The C\u2014CI, complex mol\u00adecules are linked by pairs of weak C\u2014H\u22efBr hydrogen bonds (Table\u00a01Cg1\u22efCg2iii = 3.683\u2005(1)\u2005\u00c5; Cg1 and Cg2 are the centroids of the N1/C4\u2013C8 and N2/C9\u2013C13 rings, respectively; symmetry code: (iii) 1\u00a0\u2212\u00a0x, \u2212y, 1\u00a0\u2212\u00a0z], forming a three-dimensional supra\u00admolecular structure \u2005\u00c5 and Cg5\u22efCg6iv = 4.002\u2005(2)\u2005\u00c5; Cg3, Cg4, Cg5 and Cg6 are the centroids of the C4\u2013C9, C16\u2013C21, N1/C8\u2013C12 and N2/C13\u2013C17 rings, respectively; symmetry code: (iv) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, \u2212z]. These inter\u00adactions lead to the formation of a three-dimensional network structure 3(N-N)], some structures have been reported complexes with a bidentate bi\u00adpyridine derivative ligand (N-N) of the form et al. .The ligands, debpy and debqn, were prepared as described by Chandrasekharam al. 2011 and Hoer al. 2006. The ligSynthesis of I and II: Compounds I and II were handled and stored in the dark to minimize exposure to light. For the synthesis of I, [MnBr(CO)5] and debpy were dissolved in CHCl3 (10\u2005ml). The reaction mixture was stirred at 313\u2005K for 14\u2005h under N2. After the solvent was evaporated under reduced pressure, an excess of Et2O (30\u2005ml) was added to the solution; then, the solution was allowed to stand at 253\u2005K overnight. The resultant precipitate was collected by filtration, washed with Et2O, and then dried under vacuum . Red crystals, suitable for the X-ray diffraction experiment, were grown by diffusion of n-hexane into an acetone solution of I for one week. FTIR (KBr pellet): \u03bdCO /cm\u22121 = 2028, 1918 (br) (C\u2261O), 1730 (C=O). UV\u2013vis (CHCl3): \u03bb /nm (\u220a /M\u22121 cm\u22121) = 483\u2005(3700), 367\u2005(4100), 318\u2005(21000), 247\u2005(24000).5] and debqn for 20\u2005h afforded II . Purple crystals, suitable for the X-ray diffraction experiment, were grown by diffusion of n-hexane into an acetone solution of II for one week. FTIR (KBr pellet): \u03bdCO /cm\u22121 = 2016, 1942, 1926 (C\u2261O), 1725 (C=O). UV\u2013vis (CHCl3): \u03bb /nm (\u220a /M\u22121 cm\u22121) = 548\u2005(3200), 383\u2005(19000), 276\u2005(37000).A similar reaction between [MnBr(CO)Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989020010750/dj2012sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989020010750/dj2012Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020010750/dj2012Isup4.molSupporting information file. DOI: 10.1107/S2056989020010750/dj2012IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989020010750/dj2012IIsup5.molSupporting information file. DOI: 2021226, 2021225CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II complex, a newly synthesized dye, the metal atom is coordinated by two N atoms and two O atoms from two bidentate (E)-1-[diazen\u00adyl]naphthalen-2-olate ligands.In the title Cu 16H8Br3N2O)2]\u00b7C2H6OS, the CuII atom is tetra\u00adcoordinated in a square-planar coordination, being surrounded by two N atoms and two O atoms from two N,O-bidentate (E)-1-[diazen\u00adyl]naphthalen-2-olate ligands. The two N atoms and two O atoms around the metal center are trans to each other, with an O\u2014Cu\u2014O bond angle of 177.90\u2005(16)\u00b0 and a N\u2014Cu\u2014N bond angle of 177.8\u2005(2)\u00b0. The average distances between the CuII atom and the coordinated O and N atoms are 1.892\u2005(4) and 1.976\u2005(4)\u2005\u00c5, respectively. In the crystal, complexes are linked by C\u2014H\u22efO hydrogen bonds and by \u03c0\u2013\u03c0 inter\u00adactions involving adjacent naphthalene ring systems [centroid\u2013centroid distance = 3.679\u2005(4)\u2005\u00c5]. The disordered DMSO mol\u00adecules inter\u00adact weakly with the complex mol\u00adecules, being positioned in the voids left by the packing arrangement of the square-planar complexes. The DMSO solvent mol\u00adecule is disordered over two positions with occupancies of 0.70 and 0.30.In the title compound, [Cu(C The N\u2014Cu\u2014N bond angle is 177.8\u2005(2)\u00b0. The two Cu\u2014O distances are 1.882\u2005(4) and 1.892\u2005(4)\u2005\u00c5. All bond lengths are similar to those observed in similar crystal structures shows a multiplet around 7 and 8 ppm attributed to the aromatic protons. The IR spectrum of the complex shows the vibration bands: \u03bd(N=N); 1360\u2005cm\u22121, \u03bd(C\u2014N): 1149\u2005cm \u22121, \u03bd(C\u2014Br): 645\u2005cm\u22121, \u03bd(C\u2014O): 1207\u2005cm\u22121 (aromatic), \u03bd(C=C): 1498\u2005cm\u22121 (aromatic), \u03bd(C\u2014H): 2945\u2005cm\u22121 (aromatic), \u03bd(Cu\u2014N): 417\u2005cm\u22121, \u03bd(Cu-O): 558\u2005cm\u22121. The UV\u2013Vis spectrum measured in CH2Cl2 (10 \u22125 M), shows three absorption bands: an intense band at 268\u2005nm (\u220a = 29.94 108 M\u22121 cm\u22121) attributed to intra-ligand charge-transfer transition, a band at 382\u2005nm (\u220a = 79.21 107 M\u22121 cm\u22121) associated with the azo form of the ligand and a band at 462\u2005nm (\u220a = 63.84 107 M\u22121 cm\u22121) attributed to metal\u2013ligand charge transfer.The complex, bis-1--2-naphtho\u00adlatecopper(II), was obtained by mixing 1\u2005mmol of 1--2-naphthol dissolved in 20\u2005ml of THF with 0.5\u2005mmol of Cu(OAc)Uiso(H) = 1.2 Ueq(C). An absorption correction was not applied in view of the very small size of the crystal [0.1 \u00d7 0.09 \u00d7 0.08\u2005mm]. The DMSO solvent mol\u00adecule shows disorder over two positions with final occupancies of 0.70 and 0.30. The disordered atoms were modelled as anisotropic using EADP restraints. H atoms of the disordered DMSO were omitted.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020001863/tx2018sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989020001863/tx2018Isup2.hklStructure factors: contains datablock(s) I. DOI: 1983017CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The cationic layers are made up from two types of [Na(H2O)6] octa\u00adhedra that form linear 1\u221e[Na(H2O)4/2(H2O)2/1] chains linked by dimeric [Na(H2O)2/2(H2O)4/1]2 units on both sides of the chains. The isolated trigonal\u2013pyramidal sulfite anions are connected to the cationic layers through an intricate network of O\u2014H\u22efO hydrogen bonds, together with a remarkable O\u2014H\u22efS hydrogen bond, with an O\u22efS donor\u2013acceptor distance of 3.2582\u2005(6)\u2005\u00c5, which is about 0.05\u2005\u00c5 shorter than the average for O\u2014H\u22efS hydrogen bonds in thio\u00adsalt hydrates and organic sulfur com\u00adpounds of the type Y\u2014S\u2014Z . Structural relationships between monoclinic Na2SO3(H2O)7 and ortho\u00adrhom\u00adbic Na2CO3(H2O)7 are discussed in detail.The monoclinic crystal structure of Na Indeed, the two hepta\u00adhydrates show not only a close metrical relationship , as a corrosion inhibitor in aqueous media, as a bleaching agent, as a solubilizing agent for cellulose, straw and wood in the pulp and paper industry, or as an additive in dying processes. In the USA alone, the production of sodium sulfite reached 150 000 tons in 2002 7 were grown by recrystallization of a commercial anhydrous sample from an aqueous solution at room tem\u00adper\u00adature by slow evaporation over the course of several days. In order to remove adherent mother liquor, the crystals were placed on filter paper and subsequently immersed in Paratone oil. The crystal under investigation was cleaved from a larger specimen.Colourless prismatic crystals of Na2SO3(H2O)7 was originally solved and refined in the space group P121/n1 (No. 14), with lattice parameters a = 11.8576\u2005(8), b = 7.2197\u2005(5), c = 12.6965\u2005(9)\u2005\u00c5 and \u03b2 = 106.7938\u2005(17)\u00b0 at 100\u2005K . The values for the lattice parameters at 100\u2005K are in good agreement with the previous study, with values of a = 11.922, b = 7.260, c = 12.765\u2005\u00c5 and \u03b2 = 107.22\u00b0 obtained at room temperature from Weissenberg film data 7 7, using the matrix 7 and \u03b2-Na2CO3(H2O)7. All H atoms present in the crystal structure of Na2SO3(H2O)7 were located in a difference Fourier map and were refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a022SO3(H2O)7, all atoms are located on general sites. The two sodium cations are surrounded by six water mol\u00adecules, defining a distorted octa\u00adhedral coordination polyhedron in each case. The Na\u2014O distances range from 2.3690\u2005(6) to 2.4952\u2005(6)\u2005\u00c5 (Table\u00a03[6]\u2014O of 2.44\u2005(11)\u2005\u00c5 calculated for 5520 individual bonds \u2013176.42\u2005(2)\u00b0 for Na1 and 165.81\u2005(2)\u2013174.23\u2005(2)\u00b0 for Na2, and for cis O atoms in the range 81.464\u2005(19)\u2013101.74\u2005(2)\u00b0 for Na1 and 81.51\u2005(2)\u2013103.23\u2005(2)\u00b0 for Na2. The two types of [Na(H2O)6] octa\u00adhedra show a different linkage pattern. Octa\u00adhedra centred by Na1 share common edges 4/2(H2O)2/1] chains running parallel to [001], whereas octa\u00adhedra centred by Na2 make up dimeric [Na2(H2O)2/2(H2O)4/1]2 units by sharing an edge (O5 and O5iii). In both cases, the corresponding Na\u2014O bonds to the shared O atoms at the edges are the shortest in the respective octa\u00adhedron. The dimeric units connect adjacent chains by sharing the terminal water mol\u00adecules (O9 and O7) on both sides of the chains (corner-sharing links). This way, the sodium\u2013water octa\u00adhedra are assembled by edge- and corner-sharing into an infinite layer extending parallel to (100) .In the crystal structure of Na\u00c5 Table\u00a03, with me0) Fig.\u00a01a.IV atom occupying the pyramidal position. Atom S1 is 0.5912\u2005(4)\u2005\u00c5 above the basal plane formed by atoms O1, O2 and O3. The S\u2014O bond lengths are in a narrow range 1.5224\u2005(5)\u20131.5338\u2005(5)\u2005\u00c5 [mean 1.527\u2005(6)\u2005\u00c5], just like the O\u2014S\u2014O angles . Again, these values are in good agreement with the grand mean SIV\u2014O bond length of 1.529\u2005(15)\u2005\u00c5 calculated for 90 bonds and with the O\u2014SIV\u2014O angles in the range \u223c99\u2013107\u00b0 with a mean value of \u223c104\u00b0 4/2(H2O)2/1] chains .The sulfite anion has the characteristic trigonal\u2013pyramidal configuration, with the Sns Fig.\u00a02a.a). Based on the donor\u2013acceptor distances between 2.7204\u2005(7) and 2.9110\u2005(8)\u2005\u00c5 (Table\u00a04a). Thereby, atom O1 is the acceptor of three, O2 of four and O3 of three hydrogen bonds. It is worth noting that the S\u2014O bond lengths reflect this situation nicely, with S1\u2014O2 = 1.5338\u2005(5)\u2005\u00c5 being about 0.01\u2005\u00c5 longer than the remaining two. The O9 water mol\u00adecule, bonded to Na1, Na2 and via H9A to O2, lacks a clearcut hydrogen bond for its second H atom (H9B), which points to H6B of the O6\u2014H6B\u22efO3 hydrogen bond \u00b7H2O 2]\u00b72.25H2O (S2O5) 7 7 and ortho\u00adrhom\u00adbic Na2CO3(H2O)7 (Table\u00a012O)6] octa\u00adhedra is present in the carbonate, likewise situated at x \u2243 0, b). The carbonate groups do not show pyramidalization 2/2(H2O)4/1]2 dimers.The close structural relationship between monoclinic Na7 Table\u00a01 becomes ll Fig.\u00a02b. The c2O)2/2(H2O)4/1]2 dimers in the layers. Whereas in the sulfite structure, the dimers at y \u2243 0 and y \u2243 c-glide plane 7, to NH4SO3(H2O) 6 6, which is built up from [Mg(H2O)6] octa\u00adhedra and isolated SO3 pyramids within a lattice of the space group type R3, and with Mg and S atoms both located on threefold rotation axes, there are two independent water mol\u00adecules that donate, apart from one water\u2013water hydrogen bond, three water\u2013Osulfite hydrogen bonds to each sulfite O atom, com\u00adparable to O1 and O3 in Na2SO3(H2O)7, but with shorter O\u22efO distances than in the latter. An electron deformation density study of MgSO3(H2O)6 6 nor NH4SO3(H2O) contain O\u2014H\u22efS or N\u2014H\u22efS hydrogen bonds.Crystal structures with sulfite groups anchored exclusively by hydrogen bonds are at present restricted to the title com\u00adpound Na2SO4(H2O)7 has a com\u00adpletely different arrangement of the principal building units. Its crystal structure is com\u00adprised of [Na(H2O)]6 octa\u00adhedra concatenated by edge- and corner-sharing into a three-dimensional network with isolated tetra\u00adhedral sulfate anions hydrogen bonded to the chains. Also, for sodium com\u00adpounds with analogous trigonal\u2013pyramidal oxoanions and the same charge, i.e.XO32\u2212, with X = Se and Te, no phases related structurally or com\u00adpositionally to Na2SO3(H2O)7 are known. For Na2SeO3, the anhydrous form is made up from [NaO6] octa\u00adhedra and trigonal\u2013pyramidal SeO32\u2212 anions 5 5 . These structures are based on two- or three-dimensional assemblies of [NaO5] polyhedra (Se) and [NaO6] octa\u00adhedra (Se and Te), to which SeO3/TeO3 groups are bonded via two (Se) or one (Te) O atom. The [NaO6] octa\u00adhedra in these two salts share common faces and edges but no vertices. As pointed out by Philippot et al. (19792TeO3(H2O)5 and confirmed also for Na2SeO3(H2O)5 7 structure and vice versa. A further com\u00adparison with other hydrated sodium com\u00adpounds com\u00adprised of related oxo anions shows no close structural relationship to the title hepta\u00adhydrate. For example, Na al. 1979 for Na2T10.1107/S2053229620004404/ep3004sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2053229620004404/ep3004Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2053229620004404/ep3004sup3.txtCIF with full numerical data (setting P121/n1). DOI: Click here for additional data file.10.1107/S2053229620004404/ep3004Isup4.cmlSupporting information file. DOI: 1993827CCDC reference:"} +{"text": "Within the sheets, very weak \u03c0\u2013\u03c0 stacking inter\u00adactions occur. The Hirshfeld surface analysis and fingerprint plots reveal that the crystal structure is dominated by H\u22efH (37.1%) and C\u22efH (30.1%) contacts.In the crystal, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, resulting in the formation of sheets along the 14H12INO, was synthesized by condensation of 2-hy\u00addroxy-3-methyl\u00adbenzaldehyde and 2-iodo\u00adaniline, and crystallizes in the ortho\u00adrhom\u00adbic space group P212121. The 2-iodo\u00adphenyl and benzene rings are twisted with respect to each other, making a dihedral angle of 31.38\u2005(2)\u00b0. The mol\u00adecular structure is stabilized by an O\u2014H\u22efN hydrogen bond, forming an S(6) ring motif. In the crystal, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, resulting in the formation of sheets along the a-axis direction. Within the sheets, very weak \u03c0\u2013\u03c0 stacking inter\u00adactions lead to additional stabilization. The Hirshfeld surface analysis and fingerprint plots reveal that the crystal structure is dominated by H\u22efH (37.1%) and C\u22efH (30.1%) contacts. Hydrogen bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. The crystal studied was refined as a two-component inversion twin.The title compound, C We report here the synthesis and the crystal and mol\u00adecular structures of the title compound, along with the results of a Hirshfeld surface analysis.In the present study, a new Schiff base, S(6) ring motif, which stabilizes the mol\u00adecular structure and induces the Schiff base atoms to be coplanar with the methyl\u00adphenol moiety. Of this planar unit (r.m.s deviation = 0.0274\u2005\u00c5), atoms O1 and N1 show the largest deviations from planarity in positive and negative directions [O1 = 0.035\u2005(4)\u2005\u00c5 and N1 = \u22120.060\u2005(4)\u2005\u00c5]. The C7\u2014N1 and C13\u2014O1 bonds of the title compound are the most important indicators of the tautomeric type. The C13\u2014O1 bond is of double-bond character for the keto\u2013amine tautomer, whereas this bond displays single-bond character in the enol\u2013imine tautomer. In addition, the C7\u2014N1 bond is also a double bond in the enol\u2013imine tautomer and a single bond length in the keto\u2013amine tautomer. In the title compound, the enol\u2013imine form is favored over the keto-amine form, as indicated by the C13\u2014O1 [1.352\u2005(6)\u2005\u00c5] and C7\u2014N1 [1.286\u2005(8)\u2005\u00c5] bonds, whose lengths indicate a high degree of single-bond and double-bond character, respectively. The shortest C\u2014C distance (C3\u2014C4) is 1.344\u2005(11)\u2005\u00c5 in the C1\u2013C6 ring with the weighted average ring bond distance being 1.376\u2005(11)\u2005\u00c5 for this ring.Depending on the tautomers, two types of intra\u00admolecular hydrogen bonds are observed in Schiff bases: O\u2014H\u22efN in enol\u2013imine and N\u2014H\u22efO in keto\u2013amine tautomers. Most of these compounds are non-planar. The title compound, (I)a-axis direction by C2\u2014H2\u22efCg2i inter\u00adactions (Table\u00a01Cg1\u22efCg2ii of 4.093\u2005(2)\u2005\u00c5 of the title compound are illustrated in Fig.\u00a04a and red areas on phenyl rings mapped with shape-index correspond to the H\u22ef\u03c0 contacts resulting from hydrogen bond C\u2014H\u22ef\u03c0(ring) Table\u00a01 and \u03c0\u2013\u03c0 et al., 2016E)-2-[(2-iodo\u00adphenyl\u00adimino)\u00admeth\u00adyl]phenol gave six hits: bis\u00adcopper(II) bis\u00adimino]\u00admeth\u00adyl}phenolato)bis\u00ad(iso\u00adthio\u00adcyan\u00adato)\u00addiiron(III) methanol solvate bis\u00ad(m-methano\u00adlato)tetra\u00adkis\u00adimino]\u00admeth\u00adyl}phenolato)bis\u00ad(iso\u00adthio\u00adcyanato)\u00adtetra\u00adiron(III) di\u00adchloro\u00admethane solvate in ethanol (20\u2005ml) and 2-iodo\u00adaniline in ethanol (20\u2005ml). The reaction mixture was stirred for 4\u2005h under reflux. Single crystals of the title compound for X-ray analysis were obtained by slow evaporation of an ethanol solution .Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. Hydroxyl H atoms were placed according to a difference-Fourier map and were freely refined. The crystal studied was refined as a two-component inversion twin. This reflection file contains the non-overlapping reflections of the two twin components as well as the overlapping reflections. The BASF parameter for this two-component twin refined to \u22120.03242\u2005(8).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020011974/zl2795sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020011974/zl2795Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020011974/zl2795Isup3.cmlSupporting information file. DOI: 2026445CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Alzheimer\u2019s disease (AD)-related tauopathy can be measured with CSF phosphorylated tau (pTau) and tau PET. We aim to investigate the associations between these measurements and their relative ability to predict subsequent disease progression.18F-florbetapir or 18F-florbetaben), 18F-flortaucipir (FTP) PET, CSF measurements, structural MRI, and cognition, we examined inter-relationships between these biomarkers and their predictions of subsequent FTP and cognition changes.In 219 cognitively unimpaired and 122 impaired Alzheimer\u2019s Disease Neuroimaging Initiative participants with concurrent amyloid-\u03b2 (A\u03b2) PET were 4 times more prevalent (p\u2009<\u2009\u00a00.001) than abnormal FTP only individuals (6.8%). Furthermore, among individuals on the AD pathway, CSF pTau/A\u03b240 mediates the association between A\u03b2 PET and FTP PET accumulation, but FTP PET is more closely linked to subsequent cognitive decline than CSF pTau/A\u03b240.The use of a CSF pTau/A\u03b240 may be a superior measure of tauopathy compared to CSF pTau alone, and CSF pTau/A\u03b240 enables detection of tau accumulation at an earlier stage than FTP among A\u03b2+ individuals.Together, these findings suggest that CSF pTau/A\u03b2 Extracellular amyloid-\u03b2 (A\u03b2) peptides in cortical A\u03b2 plaques and intracellular phosphorylated tau protein as neurofibrillary tangles are key hallmarks of Alzheimer\u2019s disease (AD) that can be measured in vivo with positron emission tomography (PET) imaging and biofluid markers including plasma and cerebrospinal fluid (CSF) assays. The relationship between CSF A\u03b2 and A\u03b2 PET in AD has been widely reported \u20138, but r42 in the A\u03b2\u2212 range ) associated with CSF A\u03b242 in A\u03b2 PET\u2212 participants, whereas no association was found in A\u03b2+ participants sample of 384 A\u03b2\u2212 participants and A\u03b2+ participants.CSF A\u03b2tus Fig.\u00a0a, b. Bef40 , CSF pTau/A\u03b240 , and tau PET in Temporal-metaROI and entorhinal in A\u03b2+ participants. Notably, the association with A\u03b2 PET increased from rho value 0.38 when using CSF pTau alone to 0.60 using the CSF pTau/A\u03b240 .We found A\u03b2 PET was significantly associated with CSF and PET tau measurements, which were driven by A\u03b2+ individuals. Baseline A\u03b2 PET was positively associated with CSF pTau , A\u03b2\u2212 and A\u03b2+ participants were classified as tau normal (T\u2212)/abnormal (T+) using CSF pTau or CSF pTau/A\u03b240 or tau PET thresholds, dividing the whole cohort into A\u2212/T\u2212, A\u2212/T+, A+/T\u2212, and A+/T+ groups. Few A\u03b2\u2212 participants had abnormal CSF pTau/A\u03b240 (7.6%) and temporal tau PET (5.3%), whereas A\u03b2+ participants showed a 3.0\u20134.5 times higher percentage of abnormal CSF pTau/A\u03b240 (32.6%) and temporal tau PET (24.0%) than A\u03b2\u2212 participants /abnormal (+) on CSF pTau or CSF pTau/A\u03b240 (PTAU+/\u2212) and entorhinal or Temporal-metaROI FTP SUVR (FTP+/\u2212). Abnormal CSF pTau only had higher prevalence than abnormal CSF pTau/A\u03b240 only in A\u03b2\u2212 participants, whereas abnormal CSF pTau/A\u03b240 only had marginally higher prevalence than abnormal CSF pTau only in A\u03b2+ participants. CSF pTau/A\u03b240 were positively associated with temporal tau PET across all participants. A\u03b2+ participants were responsible for this relationship because no association was found in A\u03b2\u2212 participants . The results were similar for entorhinal tau PET , CSF pTau/A\u03b240 , and Temporal-metaROI tau PET were all associated with subsequent tau PET increase (\u0394FTP) in A\u03b2+ participants . CSF pTau/A\u03b240-involved pathways explained 70% of the association between A\u03b2 PET and \u0394FTP increase in A\u03b2+ participants.The latent variable model demonstrated that the direct association between A\u03b2 and \u0394FTP increase in A\u03b2+ participants was not significant after including the CSF pTau/A\u03b2FTP Fig.\u00a0d, reduci\u03b2std\u2009=\u2009\u2212\u20090.41 ), CSF pTau/A\u03b240 , and Temporal-metaROI tau PET were all associated with subsequent cognitive decline in A\u03b2+ participants remained predictive when all variables were added into one multivariate model. The results were similar for entorhinal tau PET. In contrast, only CSF pTau/A\u03b240 was associated with subsequent cognitive decline in A\u03b2\u2212 participants.Baseline A\u03b2 PET Tau PET associations with CSF pTau/A\u03b240 were highest in medial and lateral temporal regions. (3) Associations between A\u03b2 PET, CSF pTau/A\u03b240, and tau PET were substantially driven by A\u03b2 PET-positive individuals. (4) Among these A\u03b2+ individuals, most participants (66%) were concordant on CSF pTau/A\u03b240 and Temporal-metaROI tau PET, but among discordant individuals, those with abnormal CSF pTau/A\u03b240 and normal tau PET were 4 times more prevalent (26.7%) than those with abnormal tau PET and normal CSF pTau/A\u03b240 (6.8%). (5) Among these A\u03b2+ individuals, baseline A\u03b2 PET, CSF pTau/A\u03b240, and tau PET were all associated with subsequent tau PET increase, while CSF pTau/A\u03b240 significantly mediates the association between A\u03b2 PET and tau PET . (6) Only tau PET was predictive of longitudinal cognitive decline when baseline A\u03b2 PET, CSF pTau/A\u03b240, and\u00a0tau PET were put in one multivariate model.This study had several primary findings: (1) use of a CSF pTau/A\u03b242 , but it is possible that early and late changes in CSF A\u03b240 may contribute to the tau-related effects we observed.Our motivation to adjust CSF pTau measurements was based on our observation that A\u03b2 PET-negative individuals had abnormal (\u201cpositive\u201d) CSF pTau that correlated positively with high CSF A\u03b242 Fig.\u00a0c, suggesiversity , 16, and42 alone , 17\u201319, 42 alone . However studies \u201351 have al study observed40 and tau PET were stronger in ROIs in the temporal lobe than other areas such as frontal and occipital lobes that accumulate tau in later stages of disease [40 and tau PET was primarily driven by A\u03b2 PET positivity and less influenced by clinical diagnosis in the A\u03b2\u2212 individuals, which was in line with previous reports [40 or tau PET) was rarely (5.3\u20137.9%) abnormal in the A\u03b2\u2212 range and40 ratio improves the sensitivity to detect CSF tau by adjusting for individual differences in CSF production. Furthermore, although PET and CSF measures of tau are broadly concordant in the majority (76%) of individuals when measured dichotomously, our findings support recent work [In summary, we found that the use of a CSF pTau/A\u03b2ent work indicatient work . Among aAdditional file 1:Figure\u00a0S1. The ROC analysis using the Youden index classifying 280 A\u03b2- ADNI cognitively unimpaired (CU) participants and 183 A\u03b2\u2009+\u2009ADNI MCI and AD patients as the endpoint to define the cutoff \u22651.25 for Temporal-metaROI FTP SUVR. AUC: 0.876 . Among these 463 ADNI participants, 217 (47%) participants were included in the analyses of the manuscript. Figure\u00a0S2. Histograms of Temporal-metaROI FTP SUVRs of (A) all 775 ADNI participants, (B) 280 A\u03b2- ADNI CU participants and (C) 183 A\u03b2\u2009+\u2009ADNI MCI and AD patients with tau PET scan. Red dotted line is the cutoff of Temporal-metaROI FTP SUVR 1.25. Figure\u00a0S3. The ROC analysis using the Youden index classifying 280 A\u03b2- ADNI CU participants and 183 A\u03b2\u2009+\u2009ADNI MCI and AD patients as the endpoint to define the cutoff \u22651.21 for entorhinal FTP SUVR. AUC: 0.891 . Figure\u00a0S4. Histograms of entorhinal FTP SUVRs of (A) all 775 ADNI participants, (B) 280 A\u03b2- ADNI CU participants and (C) 183 A\u03b2\u2009+\u2009ADNI MCI and AD patients with tau PET scan. Red dotted line is the cutoff of entorhinal FTP SUVR 1.21. Figure\u00a0S5. The ROC analysis using the Youden index classifying 320 A\u03b2- ADNI CU participants and 429 A\u03b2\u2009+\u2009ADNI MCI and AD patients as the endpoint to define the cutoff \u226522 for CSF p-Tau. AUC: 0.865 . Among these 749 ADNI participants, 212 (28%) participants were included in the analyses of the manuscript. Figure\u00a0S6. Histograms of CSF p-Tau of (A) all 1534 ADNI participants, (B) 320 A\u03b2- ADNI CU participants and (C) 429 A\u03b2\u2009+\u2009ADNI MCI and AD patients with CSF p-Tau measurement. Red dotted line is the cutoff of CSF p-Tau 22. Figure\u00a0S7. The ROC analysis using the Youden index classifying 169 A\u03b2- ADNI CU participants and 160 A\u03b2\u2009+\u2009ADNI MCI and AD patients as the endpoint to define the cutoff \u22650.0012 for CSF p-Tau/A\u03b240 ratio. AUC: 0.976 . Among these 329 ADNI participants, 201 (61%) participants were included in the analyses of the manuscript. Figure\u00a0S8. Histograms of CSF p-Tau/A\u03b240 for (A) all 447 ADNI participants, (B) 169 A\u03b2- ADNI CU participants and (C) 160 A\u03b2\u2009+\u2009ADNI MCI and AD patients with CSF p-Tau/A\u03b240. Red dotted line is the 0.0012 cutoff for the CSF p-Tau/A\u03b240 ratio. Figure\u00a0S9. Cross-sectional associations between CSF MASS A\u03b242 and CSF p-Tau. The vertical gray dashed line reflects the abnormal threshold of CSF p-Tau. Abbreviations: p-Tau\u2009=\u2009phosphorylated tau; A\u03b2\u2009=\u2009amyloid-\u03b2; CU\u2009=\u2009cognitively unimpaired; MCI\u2009=\u2009mild cognitive impairment; AD\u2009=\u2009Alzheimer\u2019s disease. Figure\u00a0S10. Regions with significant association between CSF P-tau and FTP tau in (A) A\u03b2+, (B) CU and (C) non-demented participants. Abbreviations: Spearman rho\u2009=\u2009Spearman\u2019s correlation coefficient; p-Tau\u2009=\u2009phosphorylated tau; A\u03b2\u2009=\u2009amyloid-\u03b2; FTP\u2009=\u200918F-flortaucipir; SUVR\u2009=\u2009standardized uptake value ratio; CU\u2009=\u2009cognitively unimpaired; MCI\u2009=\u2009mild cognitive impairment; AD\u2009=\u2009Alzheimer\u2019s disease. Figure\u00a0S11. Cross-sectional associations between A\u03b2 PET, CSF p-Tau/A\u03b240 and entorhinal tau PET. (A). Associations between baseline entorhinal tau PET and A\u03b2 PET. Associations between baseline CSF p-Tau/A\u03b240 and entorhinal tau PET in the whole cohort (B), A\u03b2- (C) and A\u03b2\u2009+\u2009(D) participants. The vertical and horizontal gray dashed lines reflect the abnormal thresholds of corresponding biomarkers in x-axis and y-axis respectively. Abbreviations: A\u03b2\u2009=\u2009amyloid-\u03b2; A\u2009=\u2009A\u03b2 PET; \u2212\u2009=\u2009negative; +\u2009=\u2009positive; AD\u2009=\u2009Alzheimer\u2019s disease; CU\u2009=\u2009cognitively unimpaired; FTP\u2009=\u200918F-flortaucipir; MCI\u2009=\u2009mild cognitive impairment. Figure\u00a0S12. Cross-sectional associations between A\u03b2 PET, CSF pTau/A\u03b240 and tau PET using alternative cutoffs. Associations between baseline A\u03b2 PET and (A) CSF pTau, (B) CSF pTau/A\u03b240 and (C) temporal tau PET. Associations between baseline CSF pTau and CSF pTau/A\u03b240 in the whole cohort (D), A\u03b2- (E) and A\u03b2\u2009+\u2009(F) participants. Associations between baseline CSF pTau/A\u03b240 and Temporal-metaROI tau PET in the whole cohort (G), A\u03b2- (H) and A\u03b2\u2009+\u2009(I) participants. The vertical and horizontal gray dashed lines reflect the abnormal thresholds of corresponding biomarkers in x-axis and y-axis respectively. Abbreviations: A\u03b2\u2009=\u2009amyloid-\u03b2; A\u2009=\u2009A\u03b2 PET; \u2212\u2009=\u2009negative; +\u2009=\u2009positive; AD\u2009=\u2009Alzheimer\u2019s disease; CU\u2009=\u2009cognitively unimpaired; FTP\u2009=\u200918F-flortaucipir; MCI\u2009=\u2009mild cognitive impairment; pTau\u2009=\u2009phosphorylated tau; PTAU\u2009=\u2009CSF pTau or CSF pTau/A\u03b240 ratio; SUVR\u2009=\u2009standardized uptake value ratio; T\u2009=\u2009CSF pTau or CSF pTau/A\u03b240 or tau PET. Figure\u00a0S13. Cross-sectional associations between A\u03b2 PET, CSF p-Tau/A\u03b240 and entorhinal tau PET using alternative cutoffs. (A). Associations between baseline entorhinal tau PET and A\u03b2 PET. Associations between baseline CSF p-Tau/A\u03b240 and entorhinal tau PET in the whole cohort (B), A\u03b2- (C) and A\u03b2\u2009+\u2009(D) participants. The vertical and horizontal gray dashed lines reflect the abnormal thresholds of corresponding biomarkers in x-axis and y-axis respectively. Abbreviations: A\u03b2\u2009=\u2009amyloid-\u03b2; A\u2009=\u2009A\u03b2 PET; \u2212\u2009=\u2009negative; +\u2009=\u2009positive; AD\u2009=\u2009Alzheimer\u2019s disease; CU\u2009=\u2009cognitively unimpaired; FTP\u2009=\u200918F-flortaucipir; MCI\u2009=\u2009mild cognitive impairment."} +{"text": "The three-mol\u00adecule aggregates are connected into a supra\u00admolecular tape by amide-N\u2014H\u22efO(amide) hydrogen bonding.In the title 1:2 co-crystal, 14H14N4O2\u00b72C7H5ClO2, comprises two half mol\u00adecules of oxalamide (4LH2), as each is disposed about a centre of inversion, and two mol\u00adecules of 4-chloro\u00adbenzoic acid (CBA), each in general positions. Each 4LH2 mol\u00adecule has a (+)anti\u00adperiplanar conformation with the pyridin-4-yl residues lying to either side of the central, planar C2N2O2 chromophore with the dihedral angles between the respective central core and the pyridyl rings being 68.65\u2005(3) and 86.25\u2005(3)\u00b0, respectively, representing the major difference between the independent 4LH2 mol\u00adecules. The anti conformation of the carbonyl groups enables the formation of intra\u00admolecular amide-N\u2014H\u22efO(amide) hydrogen bonds, each completing an S(5) loop. The two independent CBA mol\u00adecules are similar and exhibit C6/CO2 dihedral angles of 8.06\u2005(10) and 17.24\u2005(8)\u00b0, indicating twisted conformations. In the crystal, two independent, three-mol\u00adecule aggregates are formed via carb\u00adoxy\u00adlic acid-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonding. These are connected into a supra\u00admolecular tape propagating parallel to [100] through amide-N\u2014H\u22efO(amide) hydrogen bonding between the independent aggregates and ten-membered {\u22efHNC2O}2 synthons. The tapes assemble into a three-dimensional architecture through pyridyl- and methyl\u00adene-C\u2014H\u22efO(carbon\u00adyl) and CBA-C\u2014H\u22efO(amide) inter\u00adactions. As revealed by a more detailed analysis of the mol\u00adecular packing by calculating the Hirshfeld surfaces and computational chemistry, are the presence of attractive and dispersive Cl\u22efC=O inter\u00adactions which provide inter\u00adaction energies approximately one-quarter of those provided by the amide-N\u2014H\u22efO(amide) hydrogen bonding sustaining the supra\u00admolecular tape.The asymmetric unit of the title 1:2 co-crystal, C The formation of such O\u2014H\u22efN hydrogen bonding is consistent with literature precedent, which indicates a very high propensity for these hydrogen-bonding patterns between carb\u00adoxy\u00adlic acids and pyridyl entities, at least in the absence of competing supra\u00admolecular synthons hydrogen bonds are formed instead , each in a general position. Pairs of 4LH2 and CBA mol\u00adecules are connected via carb\u00adoxy\u00adlic acid-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonding, Table\u00a01i.e. 4LH2(CBA)2, as shown in Fig.\u00a01The crystallographic asymmetric unit of (I)4LH2 mol\u00adecule is centrosymmetric, the central C2N2O2 chromophore in each is strictly planar. As is usually found in these mol\u00adecules \u2005\u00c5] and C14\u2014C14ii [1.539\u2005(2)\u2005\u00c5] bond lengths are longer than usual owing to the electronegative substituents connected to both carbon atoms . The conformation of each 4LH2 mol\u00adecule is (+)anti\u00adperiplanar whereby the pyridin-4-yl residues lie to either side of the planar region of the mol\u00adecule. The dihedral angles between the respective central core and the N1- and N3-pyridyl rings are 68.65\u2005(3) and 86.25\u2005(3)\u00b0, respectively. This represents the greatest conformational difference between the 4LH2 mol\u00adecules and is emphasized in the overlay diagram of Fig.\u00a02anti, enabling the formation of intra\u00admolecular amide-N\u2014H\u22efO(amide) hydrogen bonds that complete S(5) loops, Table\u00a01As each 2 group is 8.06\u2005(10)\u00b0 for the O3-mol\u00adecule indicating a closer to co-planar mol\u00adecule than for the O5-mol\u00adecule for which the equivalent dihedral angle is 17.24\u2005(8)\u00b0. Consistent with the carb\u00adoxy\u00adlic acid assignment, the C15\u2014O3(carbon\u00adyl) bond length of 1.2172\u2005(17)\u2005\u00c5 is considerably shorter than the C15\u2014O4(hy\u00addroxy) bond of 1.3196\u2005(16)\u2005\u00c5; the bonds of the O5-benzoic acid follow the same trend with C22\u2014O5 of 1.2173\u2005(17)\u2005\u00c5 compared with C22\u2014O6 of 1.3181\u2005(16)\u2005\u00c5. As seen from Fig.\u00a02To a first approximation, the two independent CBA mol\u00adecules in (I)a axis by amide-N\u2014H\u22efO(amide) hydrogen bonding and concatenated, centrosymmetric 10-membered {\u22efHNC2O}2 synthons, Fig.\u00a03a). The tapes are consolidated into a three-dimensional architecture by pyridyl- and methyl\u00adene-C\u2014H\u22efO(carbon\u00adyl) and CBA-C\u2014H\u22efO(amide) inter\u00adactions, Fig.\u00a03b).The formation of two independent, three-mol\u00adecule aggregates has already been noted above in the crystal of (I)Crystal Explorer 17 hydrogen bonds. Analogous calculations were also performed on the symmetry expanded N1- and N3-oxalamide mol\u00adecules, hereafter 4LH2-I and 4LH2-II, respectively, and on the independent O3- and O5-chloro\u00adbenzoic acid mol\u00adecules, hereafter CBA-I and CBA-II, respectively. The dnorm distances for short contacts identified through the Hirshfeld surface analysis are given in Table\u00a02The calculation of the Hirshfeld surfaces and two-dimensional fingerprint plots were accomplished with the program dnorm maps showing red spots ranging from moderate to strong intensity are illustrated in Fig.\u00a04O\u22efN1(pyrid\u00adyl) in 3M-I, carb\u00adoxy\u00adlic-O6\u2013H6O\u22efN3(pyrid\u00adyl) in 3M-II as well as the inter\u00adactions between amide-N2\u2013H2N\u22efO2(amide) and amide-N4\u2014H4N\u22efO1(amide) in 3M-I and 3M-II, respectively, while relatively weaker inter\u00adactions with moderately to weakly intense red spots between amide-C7\u22efCl1, pyridyl-C1\u2014H1\u22efO3(carboxy\u00adlic acid), pyridyl-C2\u2014H2\u22efO3(carb\u00adoxy\u00adlic acid), methyl\u00adene-C\u2014H6A\u22efO3(carb\u00adoxy\u00adlic acid), amide-O1\u22efCl1 in 3M-I, and Cl2\u22efC14(amide), methyl\u00adene-C13\u2014H13A\u22efO5(carboxylic acid), pyridyl-C8\u2014H8\u22efO5(carb\u00adoxy\u00adlic acid), pyridyl-C9\u2014H9\u22efO5(carb\u00adoxy\u00adlic acid), Cl2\u22efO2(amide) in 3M-II are observed. As well, spots due to benzene-C27\u2014H27\u22efO1(amide) are seen, i.e. providing connections between 3M-I and 3M-II.Several dnorm maps for 3M-I and 3M-II exhibit similarity for the corresponding 4LH2 and CBA mol\u00adecules with the exception of CBA-II. Pairs of CBA-II are aligned around an inversion centre with Cl2 and H25 being directly opposite each other, ostensibly forming an eight-membered heterosynthon despite the distance being longer than the cut-off value of 2.84\u2005\u00c5 for the independent 4LH2 and CBA mol\u00adecules in (I)A\u22efO3, C7\u22efCl1, Cl2\u22efC14, O1\u22efCl1 and Cl2\u22efO2 contacts are indeed electrostatic in nature, as shown from the red (electronegative) and blue (electropositive) regions on the ESP maps despite being relatively less intense when compared to those arising from the classical hydrogen bonds.To establish the nature of the inter\u00admolecular inter\u00adactions, particularly for the weaker contacts, a mapping of the electrostatic potential (ESP) was performed over the Hirshfeld surfaces through DFT-B3LYP/6-31G basis set are comparable to the data obtained from Gaussian 16, in which Cl1, O1, Cl2 and O2 possess charges of +0.0054, \u22120.0147, +0.0054 and \u22120.0125 atomic units (a.u.), respectively; while the C7 and C14 atoms each exhibit a weak electrostatic potential charge of +0.0251 and +0.0263 a.u., respectively. Therefore, the C7\u22efCl1 and C14\u22efCl2 inter\u00adactions are dispersive in nature. On the other hand, the apparent charge complementarity between the Cl2 and H25 atoms, which align around a centre of inversion as described above, indicate the existence of an electrostatic inter\u00adaction between two CBA-II mol\u00adecules, Fig.\u00a05d).Of particular inter\u00adest is the observation that the chlorine atom inter\u00adacts with the amide-C=O residue through an electron-deficient \u03c3-hole region. To complement the ESP findings on these O\u22efCl and C\u22efCl contacts, non-covalent inter\u00adaction plots were generated for the relevant pairwise mol\u00adecules using 4LH2-I, 4LH2-II, CBA-I, CBA-II, 3M-I and 3M-II. The overall fingerprint plots for the specified mol\u00adecules/aggregates are shown in Fig.\u00a08a) and those decomposed into H\u22efO/O\u22efH/ H\u22efC/C\u22efH, H\u22efN/N\u22efH and H\u22efCl/Cl\u22efH plots are shown in Fig.\u00a08b)-(e).The two-dimensional fingerprint plots were generated in order to qu\u00adantify the close contacts for di + de distances shorter than the sum of the respective van der Waals radii of 2.61, 2.64 and 2.84\u2005\u00c5 . For 3M-I, the di + de values for the H\u22efO/O\u22efH and H\u22efN contacts are, respectively, tipped at \u223c1.98, \u223c1.95 and \u223c1.68\u2005\u00c5, and are attributed to -N2\u2014H2N\u22efO2-, -O1\u22efH4N- and -N1\u22efH4O- contacts, respectively. The analogous contacts for 3M-II are tipped at 1.95\u2005\u00c5 for -H4N\u22efO1-, \u223c1.98\u2005\u00c5 for -O2\u22efH2N- and \u223c1.64\u2005\u00c5 for -N3\u22efH6O-. For H\u22efCl/Cl\u22efH in 3M-II, the contacts are each tipped at \u223c2.80\u2005\u00c5 owing to the pair of -H25\u22efCl2- and -Cl2\u22efH25- inter\u00adactions. As for the H\u22efH and H\u22efC/ C\u22efH contacts, their di + de distances are longer than the sum of their respective van der Waals radii of 2.18 and 2.79\u2005\u00c5, and hence contribute little to the overall packing of the crystal despite providing the predominant surface contacts.The overall fingerprint plot of the individual components and the corresponding three-mol\u00adecule aggregates exhibit a paw-like profile with asymmetric spikes indicating the inter-dependency of the inter\u00admolecular inter\u00adactions between mol\u00adecules to sustain the packing. The 3M-I and 3M-II aggregates display almost identical fingerprint profiles which, upon decomposition, can be delineated into H\u22efH , H\u22efC/C\u22efH , H\u22efO/O\u22efH [21.2 and 20.7%], H\u22efCl/Cl\u22efH [7.5 and 10.8%], H\u22efN/N\u22efH [6.4 and 3.8%] and other minor contacts [10.0 and 10.7%]. A detailed analysis on the corresponding decomposed fingerprint plots shows that only the H\u22efO/O\u22efH and H\u22efN/N\u22efH contacts for both 3M-I and 3M-II as well as H\u22efCl/Cl\u22efH for 3M-II have 4LH2-I and 4LH2-II mol\u00adecules exhibit similar fingerprint profiles with only slight differences in the contact distributions. In order of dominance, these are H\u22efH (36.3% for 4LH2-I and 33.8% for 4LH2-II), H\u22efO/O\u22efH , H\u22efC/C\u22efH (21.4 and 21.2%), H\u22efN/N\u22efH (11.0 and 8.3%), H\u22efCl (1.7 and 6.1%) and other minor contacts (6.0 and 7.8%). There is no major deviations in the di + de distances cf. 3M-1 and 3M-II, with only the H\u22efO/O\u22efH as well as N\u22efH contacts being shorter than the sums of their respective van der Waals radii. Each of 4LH2-I and 4LH2-II have di + de of about 1.98\u2005\u00c5 for H\u22efO/O\u22efH and \u223c1.64\u2005\u00c5 for N\u22efH contacts.The individual X\u22efH- rather than -H\u22efX-, as evidenced most notably from the distribution for O\u22efH versus H\u22efO and Cl\u22efH vs H\u22efCl . The inclination arises due to the lack of hydrogen-bond donor atoms in the CBA-I and CBA-II mol\u00adecules, other than the carb\u00adoxy\u00adlic acid groups, so they act primarily as hydrogen-bond acceptors. Among the contacts, O\u22efH and H\u22efN for CBA-I have di + de distances of \u223c2.40 and \u223c1.64\u2005\u00c5, respectively, each being shorter than the sum of the respective van der Waals radii, while the same is true for H\u22efO/O\u22efH, H\u22efN and H\u22efCl/Cl\u22efH contact for CBA-II with di + de distances of \u223c2.38, \u223c1.62 and \u223c2.82\u2005\u00c5, respectively.As for the individual CBA-I and CBA-II mol\u00adecules, major contacts comprise H\u22efH (23.7% for CBA-I and 22.1% for CBA-II), H\u22efC/C\u22efH , H\u22efO/O\u22efH (17.7 and 17.9%), H\u22efCl/Cl\u22efH (16.8 and 17.5%), H\u22efN (5.3 and 4.4%) and other minor contacts (15.7 and 13.9%). A detailed analysis of the corresponding contacts shows all major inter\u00adactions for CBA-I and CBA-II are more inclined toward -Crystal Explorer 17 and amide-N4\u2014H4N\u22efO1(amide) hydrogen bonds has the greatest energy among all close contacts present in the crystal with an inter\u00adaction energy (Eint) of \u221261.9\u2005kJ\u2005mol\u22121. This is followed by the seven-membered heterosynthon formed between 4LH2-II and CBA-II through the carb\u00adoxy\u00adlic acid-O4\u2014H4O\u22efN1(pyrid\u00adyl) hydrogen bond with the supporting pyridyl-C\u2014H8\u22efO5(carbon\u00adyl) contact so that Eint = \u221252.0\u2005kJ\u2005mol\u22121. For the analogous contact between 4LH2-I and CBA-I but lacking the supporting pyridyl-C\u2014H\u22efO5(carbon\u00adyl) contact, it is gratifying to note the inter\u00adaction energy is correspondingly less, i.e. Eint = \u221249.4\u2005kJ\u2005mol\u22121. The inter\u00adactions between amide-C7\u22efCl1 and amide-O1\u22efCl1, summing to Eint of \u221216.6\u2005kJ\u2005mol\u22121, are also significant, as are the inter\u00adactions between methyl\u00adene-C\u2013H6A\u22efO3(amide) and pyridyl-C2\u2013H2\u22efO3(amide) with Eint = \u221215.8\u2005kJ\u2005mol\u22121. The equivalent inter\u00adactions surrounding the 4LH2-II mol\u00adecule follow the same trends and give similar energies, Table\u00a03Eint of \u22128.7\u2005kJ\u2005mol\u22121. Finally, the C27\u2014H27\u22efO1(amide) inter\u00adaction exhibits an Eint of \u221220.4\u2005kJ\u2005mol\u22121.The calculation of the inter\u00adaction energies for all pairwise inter\u00adacting mol\u00adecules was performed through Eele) as highlighted by the rod-shaped energy framework with a zigzag topology due to the combination of several strong inter\u00adactions, Fig.\u00a09a). Specifically, the combination of inter\u00adactions between 4LH2-I and CBA-I through the terminal O4\u2014H4O\u22efN1 hydrogen bonding as well as between 4LH2-II and CBA-II via O6\u2014H6O\u22efN3 and C8\u2014H8\u22efO5 inter\u00adactions leads to the formation of the core framework parallel to (101). The overall Eele of these inter\u00adactions is much greater than that associated with the ten-membered synthons formed by a combination of N2\u2014H2N\u22efO2 and N4\u2014H4N\u22efO1 hydrogen bonds as evidenced from the relatively small rod radius in the energy model of the latter inter\u00adactions, which align in a parallel fashion along the b axis, Fig.\u00a09a).The crystal of (I)2O}2 synthon along with the peripheral C7\u22efCl1/O1\u22efCl1 and C14\u22efCl2/O2\u22efCl2 inter\u00adactions which lead to a ladder-like topology, Fig.\u00a09b). The combination of the electrostatic and dispersion forces results in an enhancement of the influence of the ten-membered synthons which supersedes the energy force for the terminal carb\u00adoxy\u00adlic acid-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonds as seen in the total energy framework, Fig.\u00a09c).Apart from the electrostatic forces, the crystal is also sustained by substantial dispersion forces, which are mainly associated with the ten-membered {\u22efHNC4LH2 and mono-functional carb\u00adoxy\u00adlic acids; one exception was noted in the Chemical context. A different situation pertains when bi-functional carb\u00adoxy\u00adlic acids are employed in co-crystal formation. In these circumstances, e.g. when the carb\u00adoxy\u00adlic acid is bis\u00ad(carb\u00adoxy\u00admeth\u00adyl)urea and diglycineoxamide urea hydrogen bonds, involving both pyridyl rings, leading to three-mol\u00adecule aggregates, is an almost universal trait when co-crystals are formed between N,N\u2032-bis\u00ad(pyridin-4-ylmeth\u00adyl)oxalamide (4LH2) was prepared according to a literature procedure: m.p.: 486.3\u2013487.6\u2005K; lit. 486\u2013487\u2005K was mixed with 4-chloro\u00adbenzoic acid and the mixture was then ground for 15\u2005min in the presence of a few drops of methanol. The procedure was repeated twice. Colourless blocks were obtained through careful layering of toluene (1\u2005ml) on an N,N-di\u00admethyl\u00adformamide (1\u2005ml) solution of the ground mixture. M.p.: 456.9\u2013458.6\u2005K. IR (cm\u22121): 3211 \u03bd(N\u2014H), 3052\u20142935 \u03bd(C\u2014H), 1669\u20131604 \u03bd(C=O), 1492 \u03bd(C=C), 1419 \u03bd(C\u2014N), 794 \u03bd(C\u2014Cl).The precursor, Uiso(H) set to 1.2Ueq(C). The oxygen- and nitro\u00adgen-bound H atoms were located from a difference-Fourier map and refined with O\u2014H = 0.84\u00b10.01\u2005\u00c5 and N\u2014H = 0.88\u00b10.01\u2005\u00c5, respectively, and with Uiso(H) set to 1.5Ueq(O) or 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989020000572/hb7889sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020000572/hb7889Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020000572/hb7889Isup3.cmlSupporting information file. DOI: 1978104CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The inter\u00admolecular inter\u00adactions at play were characterized via Hirshfeld surface analysis and fingerprint plots, highlighting the evident role of C\u2014H\u22efO, C\u2014H\u22efF and C\u2014H\u22ef\u03c0 inter\u00adactions in the formation of the observed crystal structure.The structural analysis of a phenyl\u00adindolizine-based drug, namely methyl 3-(4-flurobenzo\u00adyl)-7-methyl-2-phenyl\u00adindolizine-1-carboxyl\u00adate (I) was carried out; this drug shows an inhibitory action towards 24H18FNO3, crystallizes in the monoclinic centrosymmetric space group 1/nP2 and its mol\u00adecular conformation is stabilized via C\u2014H\u22efO intra\u00admolecular inter\u00adactions. The supra\u00admolecular network mainly comprises C\u2014H\u22efO, C\u2014H\u22efF and C\u2014H\u22ef\u03c0 inter\u00adactions, which contribute towards the formation of the crystal structure. The different inter\u00admolecular inter\u00adactions have been further analysed via Hirshfeld surface analysis and fingerprint plots.The title compound, C The heterocycle is substituted at the carbon atoms C6, C7 and C8 with a meth\u00adoxy carbonyl group, a phenyl ring , and a fluoro\u00adbenzoyl ring , respectively \u2005\u00c5]. The dihedral angle between the mean plane through ring Cg3 \u2005\u00c5, 157\u00b0; C15\u22efO3iii = 3.519\u2005(4)\u2005\u00c5, 137\u00b0; symmetry codes: (ii) x, y\u00a0\u2212\u00a01, z; (iii) x, y\u00a0+\u00a01, z] and C1\u2014H1\u22ef\u03c0(C15)ii , forming ribbons along the [010] direction, as shown by the green shading in Fig.\u00a03via C11\u2014H11B\u22efF1v , C11\u2014H11A\u22ef\u03c0(C5) , C11\u2014H11C\u22ef\u03c0(C8) (CN), etc.Structural details of compounds such as CAJTAI , 4-methyl\u00adpyridine (2) , 2-bromo-1-(4-fluoro\u00adphen\u00adyl)ethan-1-one (3) , and tri\u00adethyl\u00adamine in 4.5\u2005mL of aceto\u00adnitrile were added to a 10\u2005mL microwave tube under a nitro\u00adgen atmosphere of the title compound =1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020003837/xi2021sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020003837/xi2021Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020003837/xi2021Isup3.cmlSupporting information file. DOI: 1865697CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the mol\u00adecular packing, mol\u00adecules are assembled into supra\u00admolecular layers in the 32H30N4O2S4, is generated by a crystallographic twofold axis that bis\u00adects the di\u00adsulfide bond. The mol\u00adecule is twisted about this bond with the C\u2014S\u2014S\u2014C torsion angle of 90.70\u2005(8)\u00b0 indicating an orthogonal relationship between the symmetry-related halves of the mol\u00adecule. The conformation about the imine bond [1.282\u2005(2)\u2005\u00c5] is E and there is limited delocalization of \u03c0-electron density over the CN2C residue as there is a twist about the N\u2014N bond [C\u2014N\u2014N\u2014C torsion angle = \u2212166.57\u2005(15)\u00b0]. An intra\u00admolecular hydroxyl-O\u2014H\u22efN(imine) hydrogen bond closes an S(6) loop. In the crystal, methyl\u00adene-C\u2014H\u22ef\u03c0(tol\u00adyl) contacts assemble mol\u00adecules into a supra\u00admolecular layer propagating in the ab plane: the layers stack without directional inter\u00adactions between them. The analysis of the calculated Hirshfeld surfaces confirm the importance of H\u22efH contacts, which contribute 46.7% of all contacts followed by H\u22efC/C\u22efH contacts [25.5%] reflecting, in part, the C\u2014H\u22ef\u03c0(tol\u00adyl) contacts. The calculation of the inter\u00adaction energies confirm the importance of the dispersion term and the influence of the stabilizing H\u22efH contacts in the inter-layer region.The complete mol\u00adecule of the title hydrazine carbodi\u00adthio\u00adate derivative, C S-alkyl-di\u00adthio\u00adcarbazate derivatives with heterocyclic aldehydes and ketones to form mol\u00adecules of the general formula RSC(=S)N(H)N=C(R\u2032)R\u2032\u2032, where R\u2032, R\u2032\u2032 = alkyl and aryl. These mol\u00adecules are effective ligands for a variety of metals and the motivation for complexation largely stems from the promising biological activity exhibited by the derived metal complexes \u2005\u00c5 to the other side; symmetry operation (i): 1\u00a0\u2212\u00a0x, y, z. The C1\u2014S1 bond length of 1.7921\u2005(17)\u2005\u00c5 is significantly longer than the C1\u2014S2 bond of 1.7463\u2005(17)\u2005\u00c5, which is ascribed to the S1 atom participating in the S1\u2014S1i bond of 2.0439\u2005(8)\u2005\u00c5; each C1\u2014S bond is shorter than the C9\u2014S2 bond length of 1.8308\u2005(18)\u2005\u00c5.The crystallographic asymmetric unit of (I)E-conformation), N1\u2014N2 and C2=N2 bond lengths is 1.282\u2005(2), 1.409\u2005(2) and 1.286\u2005(2)\u2005\u00c5, respectively, and suggests limited delocalization of \u03c0-electron density over this residue which is consistent with a twist about the N1\u2014N2 bond as seen in the C1\u2014N1\u2014N2\u2014C2 torsion angle of \u2212166.57\u2005(15)\u00b0. The presence of an intra\u00admolecular hydroxyl-O\u2014H\u22efN(imine) hydrogen bond, Table\u00a01i\u2014C1i torsion angle being 90.70\u2005(8)\u00b0 and the dihedral angle between the two CNS2 planes being 88.22\u2005(3)\u00b0.The sequence of C1=N1 . Layers stack along the c axis without directional inter\u00adactions between them, Fig.\u00a02b).In the crystal, the only directional contact identified in the geometric analysis of the mol\u00adecular packing employing dnorm surface, electrostatic potential and two-dimensional fingerprint plot calculations were performed for (I)Crystal Explorer 17 , i.e. near the imine-C2 and tolyl ring, centroid designated Cg1, correspond to the C2\u22efO1, C2\u22efC4 short contacts inter\u00adaction, Table\u00a01A\u22ef\u03c0(tol\u00adyl) inter\u00adaction shows up as a distinctive orange \u2018pothole\u2019 on the shape-index-mapped Hirshfeld surface, Fig.\u00a03b).The Hirshfeld surface analysis comprising a), the faint red spots appearing near the tolyl-H12, methyl\u00adene-H9B and phenol-H8 atoms correlate with the faint red spots near the sulfanyl-S1, hydrazine-N1 and tolyl-C11 atoms, and correspond to the intra-layer tolyl-C12\u2014H12\u22efS1(sulfan\u00adyl), methyl\u00adene-C9\u2014H9B\u22efN1(hydrazine) and phenol-C8\u2014H8\u22efC11(tol\u00adyl) inter\u00adactions, Table\u00a02b), with the blue and red regions corresponding to positive and negative electrostatic potentials, respectively.In the views of Fig.\u00a04de and di diagonal axes for the overall fingerprint plot, Fig.\u00a05a); those delineated into H\u22efH, H\u22efC/C\u22efH, H\u22efS/S\u22efH, H\u22efO/O\u22efH, N\u22efC/C\u22efN and H\u22efN/N\u22efH contacts are illustrated in Fig.\u00a05b)\u2013(g), respectively. The percentage contributions for the different inter\u00adatomic contacts to the Hirshfeld surface are summarized in Table\u00a03de = di \u223c2.4\u2005\u00c5, Fig.\u00a05b). The tip of this H\u22efH contact corresponds to an inter-layer H6\u22efH14 contact with a distance of 2.39\u2005\u00c5, Table\u00a02de + di \u223c2.7\u2005\u00c5 in Fig.\u00a05c). The H\u22efS/S\u22efH contacts contribute 13.6% and appear as two sharp-symmetric wings at de + di \u223c2.8\u2005\u00c5, Fig.\u00a05d). This feature reflects the intra-layer tolyl-C12\u2014H12\u22efS1(sulfan\u00adyl) inter\u00adaction, Table\u00a02de + di \u223c2.8\u2005\u00c5, Fig.\u00a05e); this separation is \u223c0.08\u2005\u00c5 longer than the sum of their van der Waals radii. Although both N\u22efC/C\u22efN and H\u22efN/N\u22efH contacts appear at de + di \u223c2.6\u20132.8\u2005\u00c5 in the respective fingerprint plots, Fig.\u00a05f) and (g), their contributions to the overall Hirshfeld surface are only 3.6 and 3.4%, respectively. The contributions from the other inter\u00adatomic contacts summarized in Table\u00a03The corresponding two-dimensional fingerprint plots for the calculated Hirshfeld surface of (I)d,p) basis set with the B3LYP function. The total energy comprises four terms: i.e. the electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies and these were calculated in Crystal Explorer 17 occurs within the intra-layer region and arises from the combination of C\u2014H\u22ef\u03c0, C\u22efO and C\u22efC short contacts as well as weak C\u2014H\u22efN/C inter\u00adactions. The second most significant energy of stabilization within the intra-layer region involves a major contribution from the tolyl-C12\u2014H12\u22efS1(sulfan\u00adyl) inter\u00adaction (dominated by Edis) with a total energy of \u221229.7\u2005kJ\u2005mol\u22121. In addition, a long-range H6\u22efH16B contact is observed within the intra-layer region with a H\u22efH separation of 2.44\u2005\u00c5.In the present analysis, the pairwise inter\u00adaction energies between the mol\u00adecules in the crystal of (I)Edis energy term also makes the major contribution to the energies of stabilization in the inter-layer region, with the separation between mol\u00adecules in the inter-layer region being H\u22efH contacts. The maximum energy is not found for the shortest H6\u22efH14 contact (\u20139.5\u2005kJ\u2005mol\u22121), Table\u00a02\u22121), each with a distance of 2.51\u2005\u00c5. Views of the energy framework diagrams down the b axis are shown in Fig.\u00a06Edis in the stabilization of the crystal.The R group, i.e. R = Me S-4-methyl\u00adbenzyl\u00addithio\u00adcarbazate (10\u2005mmol) and salicyl\u00adaldehyde (10\u2005mmol) in \u223c30\u2005ml of aceto\u00adnitrile for about 2\u2005h Uiso(H) set to 1.2Ueq(C). The O-bound H atom was located in a difference-Fourier map, but was refined with an O\u2014H = 0.84\u00b10.01\u2005\u00c5 distance restraint, and with Uiso(H) set to 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989020008762/hb7929sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020008762/hb7929Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020008762/hb7929Isup3.cmlSupporting information file. DOI: 2013050CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The nickel(II) ion in 1 is octa\u00adhedrally coordinated by the O atoms of two water mol\u00adecules, two O atoms from O-monodentate 6-fluoro\u00adnicotinate ligands and two N atoms from bridging 4,4\u2032-bi\u00adpyridine ligands.A one-dimensional nickel(II) coordination polymer with the mixed ligands 6-fluoro\u00adnicotinate (6-Fnic) and 4,4\u2032-bi\u00adpyridine , namely, [Ni(H catena-poly[[di\u00adaqua\u00adbis\u00ad(6-fluoro\u00adpyridine-3-carboxyl\u00adato-\u03baO)nickel(II)]-\u03bc-4,4\u2032-bi\u00adpyri\u00addine-\u03ba2N:N\u2032] trihydrate], {[Ni(6-Fnic)2(H2O)2]\u00b73H2O}n, (1), was prepared by the reaction of nickel(II) sulfate hepta\u00adhydrate, 6-fluoro\u00adnicotinic acid (C6H4FNO2) and 4,4\u2032-bi\u00adpyridine (C10H8N2) in a mixture of water and ethanol. The nickel(II) ion in 1 is octa\u00adhedrally coordinated by the O atoms of two water mol\u00adecules, two O atoms from O-monodentate 6-fluoro\u00adnicotinate ligands and two N atoms from bridging 4,4\u2032-bi\u00adpyridine ligands, forming a trans isomer. The bridging 4,4\u2032-bi\u00adpyridine ligands connect symmetry-related nickel(II) ions into infinite one-dimensional polymeric chains running in the [11, the polymeric chains and lattice water mol\u00adecules are connected into a three-dimensional hydrogen-bonded network via strong O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds, leading to the formation of distinct hydrogen-bond ring motifs: octa\u00admeric R88(24) and hexa\u00admeric R86(16) loops.A one-dimensional nickel(II) coordination polymer with the mixed ligands 6-fluoro\u00adnicotinate (6-Fnic) and 4,4\u2032-bi\u00adpyridine , namely, On the other hand, no metal complexes of other fluorinated analogues of nicotinate have been reported so far.Although metal complexes with nicotinate have been well-studied and documented \u00b73H2O} (1). The synthesis was carried out in a mixture of water and ethanol in the hope that the coord\u00adinated water mol\u00adecules would complete the coordination sphere around the nickel(II) ion and participate in the formation of various hydrogen-bond motifs within the hydrogen-bonded framework, along with the anti\u00adcipated lattice water mol\u00adecules. Furthermore, we wanted to explore the effect of the probable weak inter\u00admolecular inter\u00adactions involving the aromatic F atoms (for example C\u2014H\u22efF inter\u00adactions) on the assembly of the polymeric chains in the crystal packing.In this work, we report the synthesis and characterization of the first metal complex with 6-fluoro\u00adnicotinate \u2013 the one-dimensional nickel(II) coordination polymer {. In this way, a trans isomer is formed (N2i\u2014Ni1\u2014N2 = 180\u00b0) ions into an infinite one-dimensional polymeric chain extending in the [12(H2O)2]\u00b73H2O}.As the nickel(II) ion and the lattice water mol\u00adecule (atom O4) are situated on an inversion center and a twofold axis, respectively, the asymmetric unit of \u00b0) Fig.\u00a01. The 6-fon Fig.\u00a02. There acis pairs of the ligating atoms [89.65\u2005(6)\u201390.87\u2005(6)\u00b0]. The Ni1\u2014O1 bond length [2.1067\u2005(16)\u2005\u00c5] is somewhat longer than the Ni1\u2014O2 and Ni1\u2014N2 bond lengths , which is in agreement with the fact that the water mol\u00adecules are bound in the axial positions of the octa\u00adhedron. The Ni\u2014Oc (c = carboxyl\u00adate) bond lengths in 1 are comparable to those seen in the related nickel(II) complexes with 6-chloro\u00adnicotinate ligands ion is slightly distorted, as indicated by the angles for the The 4,4\u2032-bypyridine ring is not coplanar with the coordinated water mol\u00adecule atom O1, but it is rotated slightly (approximately 4\u00b0) about the Ni1\u2014N2 bond, as is evident from the torsion angles Ni1\u2014N2\u2014C7\u2014C8 [176.35\u2005(19)\u00b0] and Ni1\u2014N2\u2014C11\u2014C10 [\u2212176.03\u2005(18)\u00b0]. The values of these torsion angles ought to be 180\u00b0 in the case of coplanarity of the 4,4\u2032-bi\u00adpyridine ring and the O1 atom of the coordinated water mol\u00adecule.1 mainly features strong O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds (Table\u00a01Cg1\u22efCg1(\u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a01) = 3.8148\u2005(16)\u2005\u00c5; dihedral angle between the planes = 0.00\u2005(14)\u00b0; slippage = 1.792\u2005\u00c5 and Cg1\u22efCg2 = 3.8798\u2005(16)\u2005\u00c5; dihedral angle between the planes = 11.68\u2005(13)\u00b0; slippage = 1.917\u2005\u00c5; Cg1 and Cg2 are the centroids of the 6-fluoro\u00adnicotinate pyridine (N1/C1\u2013C5) and 4,4\u2032-bi\u00adpyridine (N2/C7\u2013C11) rings, respectively]. The strong hydrogen bonds link the polymeric chains and the lattice water mol\u00adecules into an infinite three-dimensional network. The structure can be better analysed if viewed down the [1Cg1\u22efCg1].The extended structure of s Table\u00a01 and \u03c0\u2013\u03c0 rk Fig.\u00a03. While b1 coordination polymers containing bridging 4,4\u2032-bi\u00adpyridine and pyridinedi\u00adcarboxyl\u00adate ligands, there are only two structurally similar one-dimensional nickel(II) polymers with 4,4\u2032-bi\u00adpyridine and pyridine\u00adcarboxyl\u00adate . Both the coordinated (two) and lattice (three) water mol\u00adecules were released in the same step . The two small endothermic peaks in the DSC curve (63 and 115\u00b0C) suggest that the process of the water evolution is not straightforward and that the water mol\u00adecules are differently bound in 1 (coordinated vs lattice). Indeed, the polymeric chains and lattice water mol\u00adecules are assembled into a hydrogen-bonded three-dimensional structure (see Supra\u00admolecular features). It is therefore not surprising that the release of some water mol\u00adecules affects the whole hydrogen-bonded structure and leads to its complete collapse in a single, not well-resolved thermal step. The thermal decomposition of 1 continues in a broad step (observed mass loss 56.7%) in the wide temperature range of 150\u2013570\u00b0C (without any well-defined peaks in the DSC curve), which probably corresponds to the complete degradation of 1. The remaining residue at 600\u00b0C is most probably NiO.The thermal stability of \u22121 on a Perkin\u2013Elmer Spectrum TwoTM FTIR spectrometer in the ATR mode. The PXRD trace was recorded on a Philips PW 1850 diffractometer, Cu K\u03b1 radiation, voltage 40\u2005kV, current 40\u2005mA, in the angle range 5\u201350\u00b0 (2\u03b8) with a step size of 0.02\u00b0. Simultaneous TGA/DSC measurements were performed at a heating rate of 10\u00b0C min\u22121 in the temperature range 25\u2013600\u00b0C, under a nitro\u00adgen flow of 50\u2005mL\u2005min\u22121 on an Mettler-Toledo TGA/DSC 3+ instrument. Approximately 2\u2005mg of the sample were placed in a standard alumina crucible (70\u2005\u00b5l).All chemicals for the synthesis were purchased from commercial sources and used as received without further purification. The IR spectrum was obtained in the range 4000\u2013400\u2005cm1, suitable for X-ray diffraction measurements, were obtained. These were collected by filtration, washed with their mother liquor and dried in vacuo. Yield: 0.0483\u2005g (45%). Selected IR bands (ATR) : 3351 [\u03bd(O\u2014H)], 3088 [\u03bd(C\u2014H)], 1607 [\u03bd(C=O)], 1558, 1475, 1415, 1392, 1368 .6-Fluoro\u00adnicotinic acid was dissolved in distilled water (5\u2005ml), 4,4\u2032-bi\u00adpyridine was dissolved in ethanol (2\u2005mL) and nickel(II) sulfate hepta\u00adhydrate was dissolved in distilled water (2\u2005mL). The solutions of the two ligands were first mixed together under stirring. The resulting solution was then slowly added to the nickel(II) sulfate solution under stirring. The pH of the final solution was adjusted to 7 by adding an ammonia solution dropwise. The obtained, clear solution was left to evaporate slowly at room temperature for approximately three weeks until light\u2013blue crystals of Uiso(H) = 1.2Ueq(C) for aromatic H atoms]. Water H atoms were found in difference-Fourier maps, O\u2014H distances were restrained to an average value of 0.82\u2005\u00c5 using DFIX and DANG instructions and they were refined isotropically [Uiso(H) = 1.2Ueq(O)].Crystal data, data collection and structure refinement details are summarized in Table\u00a02The highest difference peak is 0.92\u2005\u00c5 away from the O3 atom and the deepest difference hole is 0.50\u2005\u00c5 away from the Ni1 atom.10.1107/S2056989020003023/xi2024sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020003023/xi2024Isup4.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020003023/xi2024sup3.docxTGA, DSC and IR data. DOI: 1988000CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Five-minute epoxy serves as a waterproof case enabling the operation of our supercapacitors while submerged underwater and a gel electrolyte extends cycling stability to 10,000 cycles with ~90% capacitance retention.Fired brick is a universal building material, produced by thousand-year-old technology, that throughout history has seldom served any other purpose. Here, we develop a scalable, cost-effective and versatile chemical synthesis using a fired brick to control oxidative radical polymerization and deposition of a nanofibrillar coating of the conducting polymer poly (PEDOT). A fired brick\u2019s open microstructure, mechanical robustness and ~8\u2009wt% \u03b1-Fe Fired brick is a universal building material, produced by thousand-year-old technology, which throughout history has seldom served any other purpose. Here, the authors show that bricks can store energy after chemical treatment to convert their iron oxide content into conducting polymer nanofibers. This masonry building block is commonly found in various red tones and mostly comprised of fused particles of silica (SiO2), alumina (Al2O3) and hematite (\u03b1-Fe2O3)2. The red color of a brick originates from hematite, a pigment first utilized by humans 73,000 years ago4 and serving today as a low-cost naturally abundant inorganic precursor for catalysts5, magnets6, and alloys7. State-of-the-art energy storage materials are also produced from hematite. For example, FeNx, FeP, and Li5FeO4 are synthesized via anionic or cationic exchange for potassium-ion batteries, Zn\u2013air batteries, pseudocapacitors, and lithium-ion batteries11; electrochemical transformation of hematite leads to FeOOH supercapacitor anodes12.Fired brick, typically used for construction and architectural esthetics, is one of the most durable materials with a 5000-year history dating back to Neolithic China13. Chemistries enabled by hematite provide an opportunity for developing cutting-edge functionalities on a fired brick where 8\u2009wt% \u03b1-Fe2O3 content and a 3D porous microstructure afford an ideal substrate for engineering a mechanically robust electrode. Here, we develop a supercapacitor using a brick\u2019s hematite microstructure as reactant to vapor-deposit a nanofibrillar coating of the conducting polymer poly (PEDOT). Vapor-phase synthesis leads to PEDOT coatings exhibiting a high electronic conductivity14 and facile charge transfer, making it an ideal route for producing electrodes15. This synthesis utilizes a brick\u2019s open microstructure and thermal stability to permeate acid and monomer vapor through its pores at 160\u2009\u00b0C to control \u03b1-Fe2O3 dissolution and Fe3+ hydrolysis with concomitant oxidative radical polymerization.This work is inspired by our recently published rust-assisted vapor phase polymerization\u22122 and energy density of 222\u2009\u00b5Wh\u2009cm\u22122 at a current density of 0.5\u2009mA\u2009cm\u22122. This two-electrode-based measurement is collected using 1\u2009M H2SO4 aqueous electrolyte under 1\u2009V operating voltage window. To mimic a \u201cbrick-mortar-brick\u201d structure, a supercapacitor is modified using a quasi-solid-state electrolyte /1\u2009M H2SO4) that also plays the role of binder and separator. Our devices are water-resistant because they are coated with an epoxy encapsulating layer that protects them enabling charge storage at temperatures between \u221220 and 60\u2009\u00b0C. A supercapacitor is stable in ambient conditions undergoing 10,000 charge\u2013discharge cycles with ~100% coulombic efficiency and ~90% capacitance retention. Moreover, a supercapacitor brick module is produced reaching a 3.6\u2009V voltage window by connecting three devices in series.A symmetric brick-based supercapacitor shows an areal capacitance of 1.60\u2009F\u2009cm2O3 at 160\u2009\u00b0C with HCl vapor; this process liberates Fe3+ ions, promotes hydrolysis and initiates precipitation of colloidal 1D FeOOH nuclei and acid (HCl) where the former leads to oxidative radical polymerization and the latter, to acid-catalyzed polymerization . Using a HCl vapor concentration less than 4.8\u2009mM leads to an incomplete reaction because both oxidative radical polymerization and acid-catalyzed polymerization are impeded exhibiting 2\u2126 two-point probe electrical resistance and nanofibers characterized by a ~30\u2009\u00b5m length and ~190\u2009nm diameter evaporation, (2) dissolution, (3) hydrolysis, and (4) polymerization. A blue PEDOT coating is visible on a brick 4\u2009h after initiating a reaction and its thickness increases inversely proportional with electrical resistance until the end of reaction at 14\u2009h. An extended polymerization time increases the polymer coating\u2019s two-point probe electrical resistance sizes and porosities are investigated with different gravel SiO sizes anted Fig.\u00a0 that fac\u20133 with dses Fig.\u00a0. These tses Fig.\u00a0. Porositsly Fig.\u00a0. Our polsly Fig.\u00a0 and pattsly Fig.\u00a0 as demonsly Fig.\u00a0. Note th2O3 particles enables deposition of PEDOT coatings on customized substrates such as concrete road pavers weighing 249\u2009mg and carrying 6.97\u2009mg PEDOT is fabricated into an electrode with one 1\u2009cm\u2009\u00d7\u20090.5\u2009cm face exposed . The electrode exhibits a quasi-rectangular shaped three-electrode cyclic voltammogram and a capacitance of 2.61\u2009F\u2009cm\u22122 (187\u2009F\u2009g\u22121 based on PEDOT\u2019s mass) at 2\u2009mV\u2009s\u22121 in 1\u2009M H2SO4 arise due to iron species in brick and disappears as scan rate increases to 25\u2009mV\u2009s\u22121 because Faradaic processes in PEDOT are faster than those occurring from solid Fe2O3 than sodium sulfate (0.878\u2009F\u2009cm\u22122) at 25\u2009mV\u2009s\u22121 vs. Na+ (5.19\u2009\u00d7\u200910\u22128\u2009m2\u2009V\u22121\u2009s\u22121) or a lower electrical resistance caused by doping at low pH14. We probed this behavior further using electrochemical impedance spectroscopy. Nyquist plots and an equivalent circuit diagram demonstrate a significantly lower ion diffusion resistance for H2SO4 (1.7\u03a9) vs. Na2SO4 (4.6\u03a9) and minimal change in electrode material electrical resistance at low pH . Diffraction patterns remain unchanged demonstrating that most inorganic species in a brick remain unaffected by synthesis and electrochemical cycling and Al (1.97\u2009\u00b5g\u2009mL\u22121) in the electrolyte after cycling, these concentrations are equivalent to mass losses (based on the entire brick) of 0.0127 and 0.0075\u2009wt% for \u03b1-Fe2O3 and Al2O3, respectively. These results confirm that inorganic species in a brick are preserved after electrochemical cycling.To quantify dissolution of a brick\u2019s \u03b1-Fe2SO4 aqueous electrolyte encompassing two PEDOT-coated bricks and a separator and its total mass (499\u2009mg) includes 13.94\u2009mg of PEDOT. Nyquist plot shows an aggregated internal resistance of 3\u03a9 and a line with a ~45\u00b0 slope between semicircle and low-frequency domain (Warburg region) leading to a device areal capacitance of 1.59\u2009F\u2009cm\u22122 calculated using the electrode area directly in contact with separator (0.5\u2009cm2) serve as electrodes in a symmetric supercapacitor (1\u2009cm\u2009\u00d7\u20090.5\u2009cm\u2009\u00d7\u20090.5625\u2009cm) using 1\u2009M Hyte Fig.\u00a0. A devicm2) Fig.\u00a0. In our \u22122 current density in a 1\u2009V window as well as areal energy and power densities of 222 and 0.25\u2009mW\u2009cm\u22122, respectively (394\u2009\u00b5Wh\u2009cm\u22123 and 0.44\u2009mW\u2009cm\u22123 for volumetric). High power density (12.5\u2009mW\u2009cm\u22122) is obtained at a current density of 25\u2009mA\u2009cm\u22122 albeit with lowered capacitance (0.706\u2009F\u2009cm\u22122) and energy density (98\u2009\u00b5Wh\u2009cm\u22122) because ion transport in our thick electrode is limited (shown by high IR drop of 0.4\u2009V). Our device works in an extended voltage window (1.2\u2009V) resulting in cyclic voltammograms and galvanostatic charge\u2013discharge curves that retain shape during galvanostatic charge-discharge experiments at 0.5\u2009mA\u2009cmitance 0.06\u2009F\u2009cm\u22122\u2009V during2SO4 gel that binds PEDOT-coated bricks and serves as electrolyte and separator containing 13.94\u2009mg of PEDOT. In a tensile test, our electrode\u2013gel\u2013electrode structure withstands a shearing force equal to 1000 times the device\u2019s weight20 and areal energy density (121\u2009\u00b5Wh\u2009cm\u22122) originating from PEDOT-coated brick electrodes are calculated from galvanostatic charge\u2013discharge curves ; cyclic voltammograms also show capacitive behavior using a poly/1\u2009M Htor Fig.\u00a0. The gelior Fig.\u00a0. The absces Fig.\u00a0.\u22122) with ~87% capacitance retention , methanol (\u226599.8%), and hydrochloric acid (37%) are purchased from Sigma-Aldrich; sulfuric acid (AR) is purchased from Macron. The PEDOT:PSS solution (Clevios PH 1000) is purchased from Heraeus company. All chemicals are used without further purification. Platinum foil is purchased from Alfa Aesar and utilized for engineering electrode leads and Celgard 3501 membrane is used as a separator. Fired bricks are purchased from local hardware stores: The Home Depot Inc. , Lowe\u2019s Inc. and Menards Inc. . Road pavers are purchased from Menards Inc. and all construction materials used for developing electrodes are cut using a diamond saw. Red cement color (\u03b1-Fe2O3 particles) produced by NewLook Inc. is purchased from The Home Depot Inc. and serves as an oxidant source for developing chemical syntheses. Materials for making concrete include commercial-grade Quikrete Portland cement (Type I/II) , Quikrete all-purpose sand (Internet #100318450Model # 115251Store SKU #137263) and Pavestone multi-purpose patio/paver base .Chlorobenzene (99%), 3,4-ethylenedioxythiophene (97%), poly (Rs), electrode material resistance (Rm), material capacitance (Cm), double-layer capacitance (Cdl), and constant phase element (CPE). Here, Rs reflects the electrolyte ionic mobility and Rm represents the electrical resistance of the electrode. Powder X-ray diffraction spectra of brick powders pulverized by mortar and pestle are obtained in a Bruker d8 Advance X-ray diffractometer at room temperature, with a Cu K\u03b1 radiation source (\u03bb\u2009=\u20091.5406\u2009\u00c5) and LynxEyeXe detector. The sample holder is a background-free silicon disk rotating at 30\u2009rpm when collecting data with a 0.02\u00b0 scan step at 40\u2009kV and 40\u2009mA. Current\u2013voltage tests are performed on a 3D printed two-point probe station with two gold probes separated by 2\u2009mm23. Water absorption experiments are performed as described in ASTM C67/C67M-18 except brick samples are 1\u2009cm\u2009\u00d7\u20090.5\u2009cm\u2009\u00d7\u20090.28\u2009cm in size. Inductively coupled plasma mass spectrometry is performed on a Perkin Elmer ELAN DRC II ICP\u2013MS. Samples for testing are obtained from electrolytes (5\u2009mL 1\u2009M H2SO4) after three-electrode cyclic voltammetry experiments and are diluted to 1/100 with Mili-Q water before analyses. External calibration curves are obtained with IV-ICPMS-71A standard solution purchased from Inorganic Ventures, Inc.Scanning electron micrographs and energy-dispersive X-ray spectra are collected with a JEOL 7001LVF FE-SEM. Two-point probe resistance measurements are carried out using a Fluke 177 True RMS digital multimeter with 3\u2009mm distance between two probes. Thermogravimetric analysis is conducted on a Discovery TGA (TA Instruments). Cyclic voltammetry, galvanostatic charge\u2013discharge measurements and electrochemical impedance spectroscopy are performed in a BioLogic VMP3 multipotentiostat. For electrochemical impedance spectroscopy, the sinusoidal disturbance is 10\u2009mV with frequencies scanned between 100\u2009kHz and 0.1\u2009Hz. A Nyquist plot shows real impedance Z\u2032 vs. imaginary impedance \u2212Z\u2032\u2032 under a sinusoidal disturbance at the open circuit potential. Fitting of Nyquist plot using an equivalent circuit diagram contains solution resistance (2SO4 (1\u2009M) is then carried out by pipetting acid on the inner wall under vigorous stirring to prevent carbonization of poly. Stirring minimizes localized heating and is carried out for 1\u2009h resulting in a homogeneous, translucent, and colorless solution.The gel electrolyte is formulated using 1\u2009g of poly powder dissolved in 10\u2009mL deionized water under vigorous stirring at 90\u2009\u00b0C and cooled to around 50\u2009\u00b0C. Dropwise addition of 1\u2009g of concentrated HBrick is cut using a diamond saw (\u00b10.03\u2009cm error) into the following 4 different sizes: 1.00\u2009cm\u2009\u2a2f\u20090.50\u2009cm\u2009\u2a2f\u20090.28\u2009cm (for studying synthesis and electrochemistry), 1.27\u2009cm\u2009\u2a2f\u20091.27\u2009cm\u2009\u2a2f\u20090.20\u2009cm (for patterning), 2.00\u2009cm\u2009\u2a2f\u20091.00\u2009cm\u2009\u2a2f\u20091.00\u2009cm and 10.16\u2009cm\u2009\u2a2f\u20096.77\u2009cm\u2009\u2a2f\u20095.72\u2009cm . A brick is thrice washed with deionized water to remove surface dust then dried at 160\u2009\u00b0C for 1\u2009h and cooled to room temperature.The syntheses of all types of 1\u2009cm\u2009\u2a2f\u20090.5\u2009cm\u2009\u2a2f\u20090.28\u2009cm brick are performed in a 25\u2009mL Teflon-lined stainless-steel autoclave as shown in Fig.\u00a0A brick (1.27\u2009cm\u2009\u2a2f\u20091.27\u2009cm\u2009\u2a2f\u20090.20\u2009cm) is patterned using a polyimide tape mask. Synthesis is carried out at 150\u2009\u00b0C for 14\u2009h in a 125\u2009mL Teflon-lined stainless-steel autoclave containing 1\u2009mL of a 0.85\u2009M EDOT solution in chlorobenzene and 0.6\u2009mL of 12\u2009M HCl with a large brick (10.16\u2009cm\u2009\u2a2f\u20096.77\u2009cm\u2009\u2a2f\u20095.72\u2009cm). The reaction is carried out using 15\u2009mL of 12\u2009M HCl and 15\u2009mL of a 0.85\u2009M EDOT in chlorobenzene solution at 150\u2009\u00b0C for 6\u2009h Fig.\u00a0.2O3 particles to a concrete surface for 3\u2009s, then dried in air. Synthesis is performed at 150\u2009\u00b0C for 14\u2009h in a 125\u2009mL Teflon-lined stainless-steel autoclave loaded with 1\u2009mL of a 0.45\u2009M EDOT in chlorobenzene solution and 0.1\u2009mL of 12\u2009M HCl.We produce a PEDOT coating on concrete by applying \u03b1-Fe\u22121 poly/1\u2009M H2SO4) is pipetted onto two 1\u2009cm\u2009\u2a2f\u20090.5\u2009cm\u2009\u2a2f\u20090.28\u2009cm PEDOT-coated bricks (100\u2009\u00b5L each brick on the 1\u2009cm\u2009\u2a2f\u20090.5\u2009cm face). This electrolyte is allowed to impregnate for 12\u2009h at ambient conditions forming a semidry gel layer. An additional 25\u2009\u00b5L of gel electrolyte is added serving as a binder between two bricks to assemble a sandwich-type supercapacitor. A device is dried in ambient conditions for 1\u2009h before sealing with epoxy . This electrolyte is allowed to stabilize for 12\u2009h at ambient conditions ; this process is repeated for all brick electrodes resulting in a thick gel layer in 1\u2009mL of 1\u2009M HSupplementary InformationPeer Review FileDescription of Additional Supplementary FilesSupplementary Movie 1"} +{"text": "The complex has hexa\u00adgonal layers of Cs3Yb3 with THF ligands and Me3Si groups in between the layers.Reduction of (C 5:\u03b75-1-(tri\u00admethyl\u00adsil\u00adyl)cyclo\u00adpenta\u00addien\u00adyl]caesium(I)ytterbium(II)], [CsYb(C8H13Si)3(C4H8O)]n or [(THF)Cs(\u03bc-\u03b75:\u03b75-Cp\u2032)3YbII]n was synthesized by reduction of a red THF solution of (C5H4SiMe3)3YbIII with excess Cs metal and identified by X-ray diffraction. The compound crystallizes as a two-dimensional array of hexa\u00adgons with alternating CsI and YbII ions at the vertices and cyclo\u00adpenta\u00addienyl groups bridging each edge. This, based off the six-electron cyclo\u00adpenta\u00addienyl rings occupying three coordination positions, gives a formally nine-coordinate tris\u00ad(cyclo\u00adpenta\u00addien\u00adyl) coordination environment to Yb and the Cs is ten-coordinate due to the three cyclo\u00adpenta\u00addienyl rings and a coordinated mol\u00adecule of THF. The complex comprises layers of Cs3Yb3 hexa\u00adgons with THF ligands and Me3Si groups in between the layers. The Yb\u2014C metrical parameters are consistent with a 4f14 YbII electron configuration.The green compound poly[(tetra\u00adhydro\u00adfuran)\u00adtris\u00ad[\u03bc-\u03b7 However, the Cs reductions yielded polymeric complexes of general formula [Cp\u2032\u2032M(\u03bc-Cp\u2032\u2032)2Cs(THF)2]n where the Cs cation has coord\u00adin\u00adated THF and cyclo\u00adpenta\u00addienide ligands. Attempts to extend this chemistry to smaller rare-earth metals by reduction of Cp\u20323Ln showed evidence of LnII in solution; however, the reduction products were highly unstable and decomposed even at 238\u2005K.The new +2 oxidation states for the rare-earth metals Y, La, Ce, Pr, Gd, Tb, Ho, Er, and Lu were recently discovered by reduction of CpC8 Fig.\u00a01 3Yb]n, 1 (Cp\u2032 = C5H4SiMe3) was isolated by reduction of the Cp\u20323YbIII complex ][Cp\u20323YbII] (crypt = 2.2.2-cryptand), which was fully characterized as a 4f14 YbII complex, Table\u00a023Ln reduction chemistry, the difference in Ln\u22efCp(centroid) distances between the LnIII and LnII complexes provides important information on the electronic configuration of the lanthanide ion distances for reduction of 4fnLnIII ions to 4fn+1LnII ions range from 0.1 to 0.2\u2005\u00c5 distance is characteristic of a 4f13 YbIII reduction to a 4f14 YbII ion. In contrast, LnII ions with 4nf5d1 configurations where the additional electron populates a d-orbital instead of the an f-orbital have differences of only 0.02\u20130.05\u2005\u00c5 \u22efYb\u22efCp\u2032(centroid) and 109.0\u2013121.4\u00b0 Cp\u2032(centroid)\u22efCs\u22efCp\u2032(centroid) angles generate the undulation of the hexa\u00adgons shown in Fig.\u00a06The side view of these layers in Fig.\u00a061 are shorter than the 3.278 and 3.435\u2005\u00c5 Cs\u22efCp\u2032\u2032(centroid) distances in [(THF)2Cs][(\u03bc-\u03b75:\u03b75-Cp\u2032\u2032)2UII(\u03b75-Cp\u2032\u2032)]n, distances in {[(Me3Si)2NCs]2\u00b7[(C5H5)2Fe)] 0.5\u00b7(C6H5Me)}n, distances in [(THF)2Cs(\u03bc3-O)3{[Ti(C5Me5)]3-(\u03bc3-CCH2)}] 2UII(\u03b75-Cp\u2032\u2032)]n 3{[Ti(C5Me5)]3(\u03bc3-CCH2)}] , 3.197\u2005(1), and 3.268\u2005(2)\u2005\u00c5 Cs\u22efCp\u2032(centroid) distances in 1 differs from that of the [(THF)2Cs][(\u03bc-\u03b75:\u03b75-Cp\u2032\u2032)2MII(\u03b75-Cp\u2032\u2032)]n, complexes , which comprise zigzag chains of \u2013M\u2013(\u03bc-Cp\u2032\u2032)\u2013Cs\u2013(\u03bc-Cp\u2032\u2032)\u2013 repeat units with a terminal Cp\u2032\u2032 attached to M and two terminal THF ligands attached to Cs coordination like Yb in 1, but the Cs is coordinated by only two cyclo\u00adpenta\u00addienyl ligands to give a bent metallocene Cp\u2032\u20322Cs(THF)2 sub-structure with these larger rings.The extended structure of et al., 2016x moieties with various types of cyclo\u00adpenta\u00addienyl rings (Cpx): [Na(\u03bc-\u03b75:\u03b75-C5H5)3YbII]n 2YbII2(\u03bc-\u03b75:\u03b75-Cp\u2032\u2032)2]n Yb(\u03bc-I)(\u03bc-\u03b75:\u03b75-C5Me5)Yb(C5Me5)]n (Ph2Pz)(THF)]n 3YbII]n 3SmII] is similar 2UII(\u03b75-Cp\u2032\u2032)]n 2NCs]2[(C5H5)2Fe)]\u00b70.5(C6H5Me)}n 3{[Ti(C5Me5)]3(\u03bc3-CCH2)}] Li]n in THF (2\u2005mL) to excess Cs as a smear produced a green solution. This was stirred for 15\u2005min at room temperature and then layered at the bottom of a vial below an Et2O (10\u2005mL) layer for crystallization at \u221235\u00b0C. After 1\u2005d, X-ray quality dark-green crystals of [(THF)Cs(\u03bc-\u03b75:\u03b75-Cp\u2032)3YbII]n were isolated. A small number of crystals were obtained and used for crystallographic analysis. Too little sample was available for other characterization.In an argon-filled glovebox, addition of a red solution of Cp\u20325:\u03b75-Cp\u2032)3YbII]n, 1 are summarized in Table\u00a03Uiso(H) values of 1.2Ueq(C) for CH2 and aromatic hydrogens and 1.5Ueq(C) for CH3 hydrogens with C\u2014H distances of 0.99 (CH2), 0.95 (aromatic), and 0.98\u2005\u00c5 (CH3).Crystal data and structure refinement for [(THF)Cs I. DOI: 10.1107/S2056989020008051/zl2785Isup2.hklStructure factors: contains datablock(s) I. DOI: 2010185CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Lolium multiflorum L.) including a cultivar , bred to accumulate larger concentrations of Mg.Magnesium (Mg) deficiency (known as grass tetany) is a serious metabolic disorder that affects grazing ruminants. We tested whether Mg-fertiliser can increase Mg concentration of Italian ryegrasses (2.6H2O). Under field conditions, the three cultivars in the CE condition and cv. Alamo were grown at two sites, and four rates of MgSO4 fertiliser application rates (0\u2013200\u00a0kg\u00a0ha\u22121 MgO). Multiple grass cuts were taken over two-years.Under controlled environment (CE) conditions, three cultivars were grown in low-nutrient compost at six fertiliser rates (0\u20131500\u00a0\u03bcM MgClp\u2009<\u20090.05) smaller than cv. Alamo. The effect of Mg-fertiliser rate on DM yield was not significant (p\u2009\u2265\u20090.05).Grass Mg concentration increased with increasing Mg-fertiliser application rates in all cultivars and conditions. Under field conditions, cv. Bb2067 had 11\u201373% greater grass Mg concentration and smaller forage tetany index (FTI) than other cultivars across the Mg-fertiliser application rates, sites and cuts. Grass dry matter (DM) yield of cv. Bb2067 was significantly contains supplementary material, which is available to authorized users. Grazing landscapes and ruminant livestock have a dominant role in the environmental, economic and food security of many countries, especially in temperate regions. For example, in 2017/18, 72% of the UK\u2019s land area (17.5\u00a0M\u00a0ha) was utilised for agriculture; of that area the proportion of grazing land was 35% permanent grass, 22% sole rights rough grazing, 6.9% common land rough grazing, and 6.5% temporary grass DEFRA . These g\u22121 DM for cattle, and 900 and 1200\u00a0mg\u00a0kg\u22121 DM for sheep and potassium (K+) ions, which is termed as the forage tetany index (FTI) is divided by the atomic weight and multiplied by the valence of the respective elements).In livestock, ~70% of Mg is stored in the skeleton and this pool is not easily mobilised when dietary Mg intake is reduced (Martens and Stumpff FTI) Eq. (Kemp anFTI) Eq. . The risFTI) Eq. . AnnuallFTI) Eq. .1\\docume+ in the soil solution suppresses the absorption of Mg2+ by plant roots digestibility which accelerates rumen passage and reducing the ruminal absorption of Mg Suttle . This iss Elliot .\u22121 DM\u2009\u00b1\u2009SD) reported include: grass hay 1400\u2009\u00b1\u2009520, grass silage 1700\u2009\u00b1\u2009540, clover silage 2300\u2009\u00b1\u2009750, lucerne hay 1700\u2009\u00b1\u2009270, maize silage 2200\u2009\u00b1\u2009690 (Suttle \u22121 DM) in New Zealand synthetic variety called Magnet with increased grass Mg concentration was bred by the Welsh Plant Breeding Station in the 1970s containing low nutrient media . Plants were watered every other day with Hortimix standard. In total there were 18 trays, each containing 8 plants of each line, randomly assigned to the central part of the tray. These plants were surrounded by cv. RvP plants to act as guard rows.The CE experiment was conducted at the Sutton Bonington Campus of the University of Nottingham in 2016. Three cultivars, cv. Bb2067, cv. Bb2068 and cv. RvP were sown on 09 August 2016 in 576 cell trays in compost with a topping of fine horticultural grade silver sand (Dejex Supplies Ltd). The trays were transferred to a controlled environment room and watered every other day with HortiMix standard (Dejex Supplies Ltd) at 10\u00a0ml\u00a0L2.6H2O. These were applied as 1\u00a0L per tray of a liquid feed based on Hoagland\u2019s media, containing 0.25\u00a0mM KH2P04, 0.5\u00a0mM KOH, 0.75\u00a0mM MgCl2.6H2O, 0.75\u00a0mM H2SO4, 0.1\u00a0mM FeNaEDTA, 2\u00a0mM Ca(NO3)2.4H2O, 2\u00a0mM NH4NO3, 30\u00a0\u03bcM H3BO3, 10\u00a0\u03bcM MnSO4.4H2O, 1\u00a0\u03bcM ZnSO4.7H2O, 3\u00a0\u03bcM CuSO4.5H2O, and 0.5\u00a0\u03bcM Na2MoO4.2H2O. The pH was maintained at 6.4.Thirty seven days after transplanting, the plants were cut leaving ~1\u00a0cm of aerial tissue. The cut plants were then watered every other day with 6\u00a0Mg treatments at 0, 75, 188, 375, 750 and 1500\u00a0\u03bcM MgCl3 and analysed by ICP-MS as described by Thomas et al. and Edinburgh, Scotland across two years (2017\u20132018). The soil type at Aberystwyth is well drained loam over gravel in the Eutric Endoskeleti-Eutric Cambisols was added at a rate of 60\u00a0kg\u00a0ha\u22121 prior to the first cut (March 2017), and then at 100, 100, and 60\u00a0kg\u00a0ha\u22121 after cuts 1, 2, 3, respectively,\u00a0and then 35\u00a0kg\u00a0ha\u22121 after all subsequent cuts. No fertiliser was added after the final cut. The Mg fertiliser was applied in April 2017 and March 2018 as magnesium sulphate (MgSO4) at MgO equivalent rates of 0, 50, 100, and 200\u00a0kg\u00a0ha\u22121. Reagent grade\u2009\u2265\u200997% anhydrous MgSO4 was dissolved in warm water and applied with a calibrated knap sack sprayer after the first sward management cutting. Magnesium fertiliser application rates were scaled in relation to a recommendation of 50\u2013200\u00a0kg\u00a0ha\u22121 of MgO application every 3\u20134\u00a0years when exchangeable soil Mg\u2009is\u00a0<\u200926\u00a0mg\u00a0L\u22121 HNO3 and analysed using ICP-MS at the University of Nottingham were collected with an auger, using a \u201cW\u201d transect across each site to determine baseline soil physico-chemical properties. Soil samples were also collected from the 16 treatments (samples from the centre of each of four replicate plots were composited) at the beginning of June 2018, after the second Mg-fertiliser application. The baseline soil pH (in water), and exchangeable Mg, Ca and K were analysed at Lancrop Laboratory while the second-year soil pH, and exchangeable Mg, Ca and K concentrations were\u00a0analysed at the University of Nottingham. At both laboratories, a similar procedure was followed. Thus, 5\u00a0mg of <2\u00a0mm sieved soil was dissolved in 25\u00a0mL of 1\u00a0Data were compiled in MS Excel and Access. For field experiments where repeated observations were made, statistical analyses were conducted using R was addressed by the use of a linear mixed model. Two models were considered. In the first (sphericity assumption) where the correlation between the residuals for any two measures on the same unit were treated as uniform. In the second an exponential autocorrelation for successive measurements was assumed. The two models were fitted using the nlme package as shown in the Online Resource (Sup Table 2.6H2O), the grass Mg concentration was 4966\u2009\u00b1\u2009880 (cv. Bb2067), 3115\u2009\u00b1\u20091018 (cv. Bb2068), and 3889\u2009\u00b1\u2009878 (cv. RvP). At the zero Mg application rate, the grass Mg concentration was 2366\u2009\u00b1\u2009381 (cv. Bb2067), 1382\u2009\u00b1\u2009343 (cv. Bb2068), and 1629\u2009\u00b1\u2009320 (cv. RvP). There was no significant interaction effect of cultivar \u00d7 Mg application rate on grass Mg concentration (p\u2009\u2265\u20090.05) (Table Grass Mg concentration increased with increasing Mg application rate (p\u2009\u2265\u20090.05) decrease with increasing Mg application rate (Table p\u2009<\u20090.01) as shown in the Online Resource Sup Table p\u2009\u2265\u20090.05, Table The FTI did not significantly (te Table . Cultivate Table . At the \u22121 at Aberystwyth and 193\u00a0mg\u00a0L\u22121 at Edinburgh (Table p\u2009<\u20090.05) due to the application of MgSO4 (Table \u22121) for control plots was 80 at Aberystwyth and 177 at Edinburgh. The increase in the exchangeable soil Mg concentration at Aberystwyth was 20%, 39%, and 74%, and at Edinburgh was 10%, 12% and 32%, at Mg-fertiliser application rates of 50, 100 and 200\u00a0kg MgO ha\u22121, respectively. Baseline exchangeable soil Ca and K concentration, and soil pH were below the recommended optimal for forage cultivation at both sites except the one between cv. Alamo and cv. RvP as shown in the Online Resource (Sup Table \u22121), the grass Mg concentrations were 2637\u2009\u00b1\u2009506 , 1674\u2009\u00b1\u2009314 , 2009\u2009\u00b1\u2009454 , and 1907\u2009\u00b1\u2009431 . At the zero Mg-fertiliser application rate, the grass Mg concentrations were 2104\u2009\u00b1\u2009495 , 1543\u2009\u00b1\u2009423 , 1786\u2009\u00b1\u2009457 , and 1787\u2009\u00b1\u2009490 . In 2018, at the largest Mg-fertiliser application rate, the grass Mg concentrations were 2990\u2009\u00b1\u2009628 (cv. Bb2067), 2414\u2009\u00b1\u2009671 (cv. Bb2068), 2691\u2009\u00b1\u2009624 (cv. Alamo), and 2563\u2009\u00b1\u2009643 (cv. RvP). At the zero Mg-fertiliser application rate, the grass Mg concentrations were 2545\u2009\u00b1\u2009589 (cv. Bb2067), 2201\u2009\u00b1\u2009694 (cv. Bb2068), 2101\u2009\u00b1\u2009572 (cv. Alamo) and 2050\u2009\u00b1\u2009553 (cv. RVP) cultivar \u00d7 Mg-fertiliser application rate, cultivar \u00d7 cutting date, and Mg-fertiliser rate \u00d7 cutting date interaction effect on grass Mg concentration. In 2017, there was cultivar \u00d7 cutting date, and Mg-fertiliser rate \u00d7 cutting date interaction effect on grass Mg concentration. There was no significant (p\u2009\u2265\u20090.05) cultivar \u00d7 Mg-fertiliser application rate \u00d7 cutting date interaction effect on the grass Mg concentration in either year except the one between cv. Alamo and cv. RvP as shown in the Online Resource (Sup Table p\u2009<\u20090.05) affected by cultivar, cutting date, and cultivar \u00d7 cutting date interaction in 2017 and 2018, but the effect of Mg-fertiliser rate on FTI was only significant in 2018. There was no statistically significant (p\u2009\u2265\u20090.05) cultivar \u00d7 Mg-fertiliser rate, Mg-fertiliser rate \u00d7 cutting date, or cultivar \u00d7 Mg-fertiliser rate \u00d7 cutting date, interaction effect on the FTI except the one between cv. Alamo and cv. RvP as shown in the Online Resource (Sup Table \u22121), the grass Mg concentrations were 2801\u2009\u00b1\u2009521 , 1698\u2009\u00b1\u2009410 , 2122\u2009\u00b1\u2009514 and 1945\u2009\u00b1\u2009433 . At the zero Mg-fertiliser application rate the grass Mg concentrations were 2337\u2009\u00b1\u2009478 , 1354\u2009\u00b1\u2009304 (cv. Bb2068), 1732\u2009\u00b1\u2009438 and 1632\u2009\u00b1\u2009386 . In 2018, at the largest Mg-fertiliser application rate, the grass Mg concentrations were 4205\u2009\u00b1\u2009746 (cv. Bb2067), 2578\u2009\u00b1\u2009503 (cv. Bb2068), 3094\u2009\u00b1\u2009726 (cv. Alamo) and 2965\u2009\u00b1\u2009707 (cv. RvP). At the zero Mg-fertiliser application rate, the grass Mg concentrations were 3239\u2009\u00b1\u2009531 , 2103\u2009\u00b1\u2009395 (cv. Bb2068), 2526\u2009\u00b1\u2009498 (cv. Alamo) and 2437\u2009\u00b1\u2009480 (cv. RVP) affected by cultivar, Mg-fertiliser application rate and cutting date. The cultivar \u00d7 cutting date, and Mg-fertiliser application rate \u00d7 cutting date interaction effect on grass Mg concentration was significant (p\u2009<\u20090.05) in both years. There was significant (p\u2009<\u00a00.05) cultivar \u00d7 Mg-fertiliser rate \u00d7 cutting date interaction effect on grass Mg concentration in 2018 but not in 2017 except the one between cv. Alamo and cv. RvP in 2017 as shown in the Online Resource (Sup Table p\u2009<\u20090.05) affected by cultivar, Mg-fertiliser application rate and cutting date. Cultivar \u00d7 cutting date, and Mg-fertiliser rate \u00d7 cutting date interaction effect was significant (p\u2009<\u20090.05) on FTI in 2017 and 2018. There was no significant (p\u2009\u2265\u20090.05) cultivar \u00d7 Mg-fertiliser, and cultivar \u00d7 Mg-fertiliser rate \u00d7 cutting date interaction effect on FTI in both years were 21.0\u2009\u00b1\u20090.7 (cv. Bb2067), 21.0\u2009\u00b1\u20090.7 (cv. Bb2068), 24.5\u2009\u00b1\u20092 (cv. Alamo), and 22.9\u2009\u00b1\u20091.0 (cv. RvP) in 2017. At the largest Mg-fertiliser application rate (200\u00a0kg\u00a0ha\u22121), total dry matter yields were 21.0\u2009\u00b1\u20090.9 (cv. Bb2067), 21.2\u2009\u00b1\u20090.9 (cv. Bb2068), 24.1\u2009\u00b1\u20090.5 (cv. Alamo), and 23.3\u2009\u00b1\u20090.9 (cv. RvP). In 2018, at the zero Mg-fertiliser application rate, total dry matter yields were 6.6\u2009\u00b1\u20090.6 (cv. Bb2067), 6.4\u2009\u00b1\u20090.5 (cv. Bb2068), 8.7\u2009\u00b1\u20090.7 (cv. Alamo), and 8.2\u2009\u00b1\u20090.8 (cv. RvP). At the largest Mg-fertiliser application rate, total dry matter yields were 6.9\u2009\u00b1\u20090.3 (cv. Bb2067), 6.7\u2009\u00b1\u20090.5 (cv. Bb2068), 9\u2009\u00b1\u20090.6 (cv. Alamo), and 9.1\u2009\u00b1\u20090.3 (cv. RvP) were 19.8\u2009\u00b1\u20090.5 (cv. Bb2067), 19.6\u2009\u00b1\u20090.9 (cv. Bb2068), 21.8\u2009\u00b1\u20091.6 (cv. Alamo), and 19.9\u2009\u00b1\u20091.5 (cv. RvP). At the largest Mg-fertiliser application rate, total dry matter yields were 19.6\u2009\u00b1\u20091.4 (cv. Bb2067), 19.7\u2009\u00b1\u20090.7 (cv. Bb2068), 20.4\u2009\u00b1\u20090.6 (cv. Alamo), and 20.2\u2009\u00b1\u20091.6 (cv. RvP). In 2018, at the zero Mg-fertiliser application rate, total dry matter yields were 7.8\u2009\u00b1\u20090.2 (cv. Bb2067), 8\u2009\u00b1\u20090.1 (cv. Bb2068), 9.4\u2009\u00b1\u20090.8 (cv. Alamo), and 9.3\u2009\u00b1\u20090.3 (cv. RvP). At the largest Mg-fertiliser application rate, dry matter yields were 7.5\u2009\u00b1\u20091 (cv. Bb2067), 8.3\u2009\u00b1\u20090.2 (cv. Bb2068), 9.1\u2009\u00b1\u20090.1 (cv. Alamo), and 8.4\u2009\u00b1\u20090.4 (cv. RvP) interaction effects of cultivar \u00d7 Mg-fertiliser application rate, cultivar \u00d7 cutting date, cultivar \u00d7 Mg-fertiliser or cultivar \u00d7 Mg-fertiliser rate \u00d7 cutting date on the grass biomass yield at both sites in both years. There was a highly significant (p\u2009<\u20090.01) negative correlation between grass biomass yield and Mg concentration. The correlation between grass DM yield and Mg concentration at Aberystwyth was , and at Edinburgh was .There were no significant (4) application rate for all the cultivars in 2017 and 2018 more S in its biomass than the other three Italian ryegrass cultivars. Planned contrasts among the cultivars mean S concentration were significant (p\u2009<\u20090.01) except the one between cv. Alamo and cv. RvP as shown in the Online Resource (Sup Table p\u2009<\u20090.05) affected by cultivar, Mg-fertiliser application rate, cutting date, and by the interaction effect of cultivar \u00d7 cutting date and Mg-fertiliser rate \u00d7 cutting date, in 2017 and 2018 (Table p\u2009<\u20090.05) affected by cultivar, Mg-fertiliser application rate and cutting date. The cultivar \u00d7 Mg-fertiliser application rate, and cultivar \u00d7 cutting date interaction effect on grass S concentration was significant (p\u2009<\u20090.05) in 2018 but not in 2017 (Table p\u2009<\u20090.05) in both years. There was no significant (p\u2009\u2265\u20090.05) cultivar \u00d7 Mg-fertiliser application rate \u00d7 cutting date interaction effect on grass S concentration , the grass S concentrations were 3257\u2009\u00b1\u2009703 (cv. Bb2067), 2738\u2009\u00b1\u2009476 , 2790\u2009\u00b1\u2009697 (cv. Alamo) and 2838\u2009\u00b1\u2009595 (cv. RvP). At the zero Mg-fertiliser application rate the grass S concentrations were 2814\u2009\u00b1\u2009588 , 2540\u2009\u00b1\u2009512 (cv. Bb2068), 2599\u2009\u00b1\u2009561 (cv. Alamo) and 2662\u2009\u00b1\u2009573 (cv. RvP). In 2018, at the largest Mg-fertiliser application rate, the grass S concentrations were 3970\u2009\u00b1\u2009845 (cv. Bb2067), 3266\u2009\u00b1\u2009592 (cv. Bb2068), 3345\u2009\u00b1\u2009429 (cv. Alamo) and 3382\u2009\u00b1\u2009604 (cv. RvP). At the zero Mg-fertiliser application rate, the grass S concentrations were 3309\u2009\u00b1\u2009601 , 3155\u2009\u00b1\u2009575 (cv. Bb2068), 2861\u2009\u00b1\u2009637 (cv. Alamo) and 2830\u2009\u00b1\u2009639 (cv. RVP) , the grass S concentrations were 4063\u2009\u00b1\u2009844 , 3153\u2009\u00b1\u2009600 , 3510\u2009\u00b1\u2009860 and 3372\u2009\u00b1\u2009782 . At the zero Mg-fertiliser application rate the grass S concentrations were 3393\u2009\u00b1\u20091033 , 2638\u2009\u00b1\u2009819 (cv. Bb2068), 2973\u2009\u00b1\u2009898 and 2931\u2009\u00b1\u2009904 (cv. RVP). In 2018, at the largest Mg-fertiliser application rate, the grass S concentrations were 4303\u2009\u00b1\u2009908 (cv. Bb2067), 3292\u2009\u00b1\u2009813 (cv. Bb2068), 3526\u2009\u00b1\u2009909 (cv. Alamo) and 3502\u2009\u00b1\u2009820 (cv. RVP). At the zero Mg-fertiliser application rate, the grass S concentrations were 3154\u2009\u00b1\u2009747 , 2885\u2009\u00b1\u2009645 (cv. Bb2068), 2835\u2009\u00b1\u2009627 (cv. Alamo) and 2742\u2009\u00b1\u2009614 (cv. RVP) for the field experiment. There are a range of potential Mg resources including dolomitic limestone of cv. Bb2067 in these trials in 2017 was well above that perennial ryegrass under conservation sward management and comparable to cv. Alamo under farmer management. There is a need to transfer cv. Bb2067 Mg-accumulating trait into the cultivars that yield larger biomass ratio in lush grasses with large N in spring during lambing season when animal requirement for S increases ESM 2(XLSX 149\u00a0kb)ESM 3(XLSX 14\u00a0kb)ESM 4(XLSX 12\u00a0kb)ESM 5(XLSX 9\u00a0kb)ESM 6(XLSX 12\u00a0kb)"} +{"text": "The structure of the title compound is stabilized by the presence of N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. Other inter\u00adactions such as C\u2014H\u22ef\u03c0 are also important in the analysis of the Hirshfeld surface. Mol\u00adecular docking studies show this compound to be a potential anti\u00adcoagulant agent. 21H17N3O5, consists of three rings, A, B and C, linked by amide bonds with the benzene rings A and C being inclined to the mean plane of the central benzene ring B by 2.99\u2005(18) and 4.57\u2005(18)\u00b0, respectively. In the crystal, mol\u00adecules are linked via N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming fused R22(18), R34(30), R44(38) rings running along [R33(37) and R33(15) rings along [001]. Hirshfeld analysis was undertaken to study the inter\u00admolecular contacts in the crystal, showing that the most significant contacts are H\u22efO/O\u22efH (30.5%), H\u22efC/C\u22efH (28.2%) and H\u22efH (29.0%). Two zones with positive potentials and two zones with negative potentials promote the N\u2014H\u22efO inter\u00adactions in the crystal. An evaluation of the mol\u00adecular coupling of the title compound and the protein with enzymatic properties known as human coagulation factor Xa (hfXa) showed the potential for coupling in three arrangements with a similar minimum binding energy, which differs by approximately 3 kcal mol\u22121 from the value for the mol\u00adecule Apixaban, which was used as a positive control inhibitor. This suggests the title compound exhibits inhibitory activity.The title compound, C Some amino\u00adbenzamide derivatives present high efficiencies in their non-linear optical properties and C form dihedral angles of of 2.99\u2005(18) and 4.57\u2005(18)\u00b0, respectively, with the mean plane of the central ring B . In turn, the amide groups C8/N2/C7(O3)/C1 (r.m.s deviation = 0.0081\u2005\u00c5) and C15/N3/C14(O4)/C10 (r.m.s deviation = 0.0101\u2005\u00c5), which link the rings, are inclined by 37.72\u2005(15) and 29.35\u2005(16)\u00b0, respectively, to ring B. The nitro group forms an angle of 5.3\u2005(2)\u00b0 with ring A while the meth\u00adoxy group is approximately coplanar with ring C, forming an angle of 1.1\u2005(5)\u00b0. The mol\u00adecule is formed by three main planes resulting from the planes that form the rings with the B ring resembling a fallen step between A and C. All bond lengths \u2212x\u00a0+\u00a01, y\u00a0+\u00a0z\u00a0+\u00a01; (ii) \u2212x, y\u00a0+\u00a0z\u00a0+\u00a01] form fused a). In turn, playing the role of complementary inter\u00adactions in crystal growth, other C\u2014H\u22efO-type inter\u00adactions link the mol\u00adecules at their ends, ensuring their stability in the growth process. Together with the N\u2014H\u22efO inter\u00adactions, they contribute to the formation of additional fused b). Weak C21\u2014H21B\u22efCg1 inter\u00adactions occur, where Cg1 is the centroid of the C1\u2013C6 benzene ring, with a H21B\u22efCg1 distance of 2.83\u2005\u00c5.The crystalline packing in compound (I)CrystalExplorer program . The overall two-dimensional fingerprint plot , and those delineated into by H\u22efO/O\u22efH, H\u22efH, H\u22efC/C\u22efH, C\u22efC, N\u22efC/C\u22efN, H\u22efN/N\u22efH, O\u22efO, O\u22efC/C\u22efO, and N\u22efO/O\u22efN contacts are illustrated in Fig.\u00a04b\u2013j, respectively, together with their relative contributions to the Hirshfeld surface. The most important inter\u00adaction corresponds to H\u22efO/O\u22efH contributing 30.5% to the overall crystal packing, as shown in Fig.\u00a04b. The pair of spikes in the fingerprint plot have a symmetrical distribution of high-density points with the tip at de + di = 1.98\u2005\u00c5. The H\u22efH inter\u00adactions, shown in Fig.\u00a04c, contribute 29.0% to the total crystal packing and appear as widely dispersed points of high density due to the large hydrogen-atom content of the mol\u00adecule with the rounded tip at de + di = 1.20\u2005\u00c5. The presence of C\u2014H\u22ef\u03c0 inter\u00adactions is shown as a pair of characteristic wings on the fingerprint plot , delineated into H\u22efC/C\u22efH contacts (28.2% contribution to the HS) having the tip at de + di = 2.84\u2005\u00c5. These results reveal the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play major roles in the crystal packing using the ot Fig.\u00a04a, and tot Fig.\u00a04d, delinGauss09W level of theory, to obtain a qualitative analysis and the Multiwfn 3.6 program for a qu\u00adanti\u00adtative analysis of (I)Autodock Vina 4.2.6 d Table\u00a02. Compound Table\u00a02, which det al., 20162-azan\u00adyl)phen\u00adyl](phenyl-\u03bb2-azan\u00adyl)methanone as the main skeleton gave 76 hits. Seven structures containing the [3-(benzoyl-\u03bb2-azan\u00adyl)phen\u00adyl](phenyl-\u03bb2-azan\u00adyl)methanone framework with nitro and meth\u00adoxy groups as substituents similar to the title compound were found, viz., N-{3-[N\u2032-(2-meth\u00adoxy\u00adphen\u00adyl)carbamo\u00adyl]-5-methyl-2-meth\u00adoxy\u00adphen\u00adyl}-2-meth\u00adoxy-5-methyl-3-nitro\u00adbenzamide carbamo\u00adyl]-5-methyl-2-meth\u00adoxy\u00adphen\u00adyl}-2-hy\u00addroxy-5-methyl-3-nitro\u00adbenzamide amino]\u00adbenzo\u00adyl}amino)-2-meth\u00adoxy\u00adbenzoate]aqua\u00adsodium methanol solvate] benzamide (ii) , previously synthesized, was subjected to an acyl\u00adation reaction in 3-nitro\u00adbenzoyl chloride (a) under chloro\u00adform reflux conditions to obtain compound (I)3-Amino-\u22121) = 3338, 3285 (N\u2014H), 3076, 2969, 1650, 1637 (C=O), 1585 (C=C), 1526, 1510, 1434, 1353 (NO2), 1316, 1301, 1241, 1137, 1026, 898, 829, 819, 721, 683\u2005cm\u22121. The UV\u2013Vis spectrum for (I)M) was obtained in DMF with a maximum of 273\u2005nm (0.62 absorption). MS , m/z [M+] calculated for C21H17N3O5+: 391.12 (100%) and 392.12 (23.1%); found: 391.10 and 392.10 (25.1%).FT\u2013IR (ATR): \u03c5 (cmUiso(H) = 1.2Ueq(parent atom) or 1.5Ueq(C). After refinement, ROTAX suggested twinning by a 180\u00b0 about [100]. This law was used to generate an hklf5 format reflection file. Refinement against this file improved R factors and residual Q peaks\u00b7BASF refined to 0.090\u2005(4)Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989020013730/dj2013sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020013730/dj2013Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989020013730/dj2013sup3.pdfSynthesis and characterization of (I). DOI: Click here for additional data file.10.1107/S2056989020013730/dj2013Isup4.cmlSupporting information file. DOI: 2016019CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The phenyl rings in the [C24H38N4O2Te2]2+ cation are in a cis arrangement to enable this species to participate in Te\u22efCl cation\u2013anion inter\u00adactions. There are also C\u2014H\u22efO inter\u00adactions involving the dimethyl sulfoxide ligands and numerous cation\u2013anion and anion\u2013anion C\u2014H\u22efCl inter\u00adactions, which link the ions into a complex three-dimensional array.In the title salt, di-\u03bc-oxido-bis\u00ad{2,6-bis\u00ad[(di\u00admethyl\u00adamino)\u00admeth\u00adyl]phenyl-\u03ba This might be due to the formation of the highly stable tellurenium(II) cation where the Te is T-shaped and involved in a three-centered, four-electron bond. While checking the reproduc\u00adibility of the reaction, in one instance, because of the adventitious uptake of oxygen, the reaction unexpectedly resulted in the isolation of the title compound, which contains the ditelluroxonium(IV) cation 2, 2 with the PdCl3(DMSO) anion. It is worth noting that Furukawa and coworkers have reported a similar diorganotelluroxonium(IV) cation namely, 2\u00b7PF6, by the reaction of the diorganotelluride 2Te with the oxidizing agent NOPF6 ]\u22122, 3, crystallizes in the triclinic space group P2 there is a cis arrangement of the aryl rings of the attached 2,6-[(di\u00admethyl\u00adamino)\u00admeth\u00adyl)]phenyl substituents 2C6H3Te(\u03bc-O)]2+\u00b7PF6\u2212, wherein the cation lies on a center of inversion and thus the aryl groups are in a trans configuration \u20132.495\u2005(2)\u2005\u00c5 for 2, are in good agreement with the values observed in 2+\u00b7PF6\u2212 [2.475\u2005(5)\u20132.486\u2005(5)\u2005\u00c5] \u00b0 and those between the Te2O2 plane and the aryl rings are 88.77\u2005(8) and 85.00\u2005(8)\u00b0, indicating that the two aryl groups are not coplanar, and are too far apart to form \u03c0\u2013\u03c0 stacking inter\u00adactions (the closest contact is between C1 and C1A at 3.672\u2005\u00c5). Thus, the driving force for the adoption of this sterically unfavorable cis conformation appears to be the formation of Te\u22efCl cation\u2013anion inter\u00adactions, which would not be possible if the trans conformation were adopted. In this case, there is a short Te2\u22efCl3 contact of 3.386\u2005(1)\u2005\u00c5 and longer contacts of 3.833\u2005(1)\u2005\u00c5 (Te2\u22efCl2) and 3.991\u2005(1)\u2005\u00c5 (Te1\u22efCl5) 2C6H3Te(\u03bc-O)]2+\u00b7PF6\u2212, no such cation\u2013anion inter\u00adactions are present and hence the more sterically favorable trans conformation is adopted. In the other two related structures containing the Te2O22+ core dication, the same cis configuration is adopted to allow the formation of inter\u00adionic Te\u22efO inter\u00adactions cation and key geometrical data are listed in Table\u00a01ts Fig.\u00a01. This isIn addition to the Te\u22efCl cation\u2013anion inter\u00adactions mentioned above, there are also C\u2014H\u22efO inter\u00adactions involving the DMSO ligands and numerous cation\u2013anion and anion\u2013anion C\u2014H\u22efCl inter\u00adactions Table\u00a02, which l2O22+ core. The first report on the mol\u00adecular structure of a diorganotelluroxonium(IV) cation was made by Furukawa and co-workers 2C6H3Te(\u03bc-O)]2+ charge-balanced as the PF6\u2212 salt. Beckmann and coworkers reported the mol\u00adecular structure of [(6-Ph2P(O)-Ace-5-) Te(\u03bc-O)]2\u00b72OTf +: 336.0545, found: 336.0541.Yield: 59%; m.p. 444\u2013446\u2005K; Ueq(C) [1.5Ueq(CH3)] and C\u2014H distances ranging from 0.95 to 0.99\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989020011482/hb7941sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020011482/hb7941Isup2.hklStructure factors: contains datablock(s) I. DOI: 1563166CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Complexes 1 and 2 both contain the \u2018diamond core\u2019 motif found previously in a number of iron, copper, and manganese high-valent bis-oxo compounds. The flexibility in the propyl linker in the ligand scaffold of 2, as compared to that of the ethyl linker in 1, results in more elongated Mn\u2014N bonds, as one would expect. The Mn\u2014Mn distances and Mn\u2014O bond lengths support an MnIV oxidation state assignment for the Mn ions in both 1 and 2. The angles around the Mn centers are consistent with the local pseudo-octa\u00adhedral geometry.The addition of The isolation and crystallographic characterization of the bis-oxo complexes 1 and 2 . The Mn ion of 1 is in a pseudo-octa\u00adhedral environment, with small deviations in the O\u2014Mn\u2014N angles relative to an ideal octa\u00adhedral geometry: O1\u2014Mn1\u2014N1 = 93.76\u2005(12), O1\u2014Mn1\u2014N2 = 92.13\u2005(12), O1\u2014Mn1\u2014N3 = 174.90\u2005(12), and O1\u2014Mn1\u2014N4 = 95.77\u2005(12)\u00b0. As is true for all diamond cores, the O1\u2014Mn1\u2014O1\u2032 angle is slightly compressed at 85.53\u2005(12)\u00b0. Metrical parameters, Mn1\u2014O1 = 1.829\u2005(3)\u2005\u00c5 and Mn1\u2014O1\u2032= 1.835\u2005(3)\u2005\u00c5 \u2005\u00c5] is slightly longer than that involving the secondary amine [Mn1\u2014N3 = 2.111\u2005(4)\u2005\u00c5]. The Mn1\u22efMn1\u2032 separation of 2.6899\u2005(15)\u2005\u00c5, falls within the normal range (2.6 to 2.8\u2005\u00c5) for bis-oxo-bridged MnIVMnIV dimers containing a diamond core. Complex 1 crystallizes with two crystallographically equivalent tetra\u00adphenyl\u00adborate counter-ions and two crystallographically equivalent water mol\u00adecules. The water mol\u00adecule is disordered over two sites with site occupancies refined to 0.870\u2005(12) and 0.130\u2005(12) for O2 and O2B respectively, with the applied constraint that both together give 100% occupancy.Complex \u00c5 Table\u00a01 fall wit2 also sits on an inversion center , making the two Mn atoms crystallographically equivalent. There is disorder in the position of the propyl linker carbon atoms . The site occupancies of N3, C1\u2013C3 and N3B, C1B\u2013C3B refined to 0.804\u2005(5) and 0.196\u2005(5), respectively, with the constraint of both together giving 100% occupancy. The Mn ion of 2 is again in a pseudo-octa\u00adhedral environment, with small deviations in O\u2014Mn\u2014N angles relative to ideal octa\u00adhedral geometry: O1\u2014Mn1\u2014N1 = 106.39\u2005(7), O1\u2014Mn1\u2014N2 = 174.90\u2005(7), O1\u2014Mn1\u2014N3 = 89.11\u2005(13), and O1\u2014Mn1\u2014N4 = 103.70\u2005(6)\u00b0. Again, as is true for all diamond cores, the O1\u2014Mn1\u2014O1\u2032 angle of 2 is slightly compressed at 85.98\u2005(7)\u00b0, and is similar to that in 1. Metrical parameters, Mn\u2014O1 = 1.8325\u2005(15) and Mn\u2014O1\u2032 = 1.8349\u2005(15)\u2005\u00c5, are also similar to those found in 1, and fall within the reported range (1.8 to 1.9\u2005\u00c5) for oxo-bridged MnIV complexes. The pyridine nitro\u00adgen atoms are once again further from the Mn ions than expected for a formal Mn\u2014N bond, but are oriented towards Mn at distances of Mn1\u2014N1 = 2.3251\u2005(18)\u2005\u00c5 and Mn1\u2014N4 = 2.3522\u2005(18)\u2005\u00c5. This bond elongation is likely to be due to steric inter\u00adference from the methyl groups at the 6-position of the pyridine rings. The nitro\u00adgens on the amine arms are much closer to the Mn center, and fall within the normal Mn\u2014N range (1.9 to 2.1\u2005\u00c5) for MnIV. The Mn\u2014N distance involving the tertiary amine [Mn1\u2014N2 = 2.1828\u2005(18)\u2005\u00c5] is noticeably longer than that involving the secondary amine [Mn1\u2014N3= 2.133\u2005(6)\u2005\u00c5]. The large difference between these bond lengths in 2, relative to those of 1, likely reflects the increased flexibility of the propyl linker in 2. The Mn1\u2014Mn1\u2032 distance [2.6825\u2005(7)\u2005\u00c5] in 2 is essentially the same as that found in 1, and falls within the normal range (2.6 to 2.8\u2005\u00c5) for bis-oxo-bridged MnIVMnIV dimers containing a diamond core. Complex 2 crystallizes with two tetra\u00adphenyl\u00adborate counter-ions and two diethyl ether mol\u00adecules per cation.Complex 1 and 2 are analogous to other reported MnIVMnIV(\u03bc-O)2 dimers. The Mn1\u2014Mn1\u2032 distances of 2.6899\u2005(15)\u2005\u00c5 in 1 and 2.6825\u2005(7)\u2005\u00c5 in 2 are comparable to other literature examples and 1.8325\u2005(15)\u2005\u00c5 for 2 are also similar to literature reported values for MnIVMnIV(\u03bc-O)2 dimers , MeCN (CaH2), and CH2Cl2 (CaH2) were dried and distilled prior to use. Et2O was rigorously degassed and purified using solvent purification columns housed in a custom stainless steel cabinet and dispensed by a stainless steel Schlenk-line (GlassContour). Complexes 3 and 4 were synthesized as described by Coggins et al. or 1.5Ueq(C-meth\u00adyl). For the disordered water mol\u00adecule in complex 1, the water was set-up as a rigid group free to rotate and move during refinement, with DFIX restraints between O and H and between both H per water. The displacement parameters of O2 and O2B were made the same with the EADP constraint. Hydrogen-atom isotropic displacement parameters were fixed at 1.5 times that of the water oxygen atoms. For the disorder in complex 2, the geometry of both groups was set to be similar with the \u2018SAME\u2019 option. Displacement parameters of N3-N3B, C1-C1B, C2-C2B, and C3-C3B were restrained with the SIMU command at 0.005 strength.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020004557/cq2034sup1.cifCrystal structure: contains datablock(s) global, Complex1, Complex2. DOI: 10.1107/S2056989020004557/cq2034Complex1sup4.hklStructure factors: contains datablock(s) Complex1. DOI: 10.1107/S2056989020004557/cq2034Complex2sup5.hklStructure factors: contains datablock(s) Complex2. DOI: 1994292, 1994291CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Enzyme promiscuity has important implications in the field of biocatalysis. In some cases, structural analogues of simple metabolic building blocks can be processed through entire pathways to give natural product derivatives that are not readily accessible by chemical means. In this study, we explored the plasticity of the aurachin biosynthesis pathway with regard to using fluoro\u2010 and chloroanthranilic acids, which are not abundant in the bacterial producers of these quinolone antibiotics. The incorporation rates of the tested precursor molecules disclosed a regiopreference for halogen substitution as well as steric limitations of enzymatic substrate tolerance. Three previously undescribed fluorinated aurachin derivatives were produced in preparative amounts by fermentation and structurally characterized. Furthermore, their antibacterial activities were evaluated in comparison to their natural congener aurachin D. Room for a halogen: The plasticity of the aurachin biosynthetic pathway in the myxobacterium S. erecta was explored for the use of fluoro\u2010 and chloroanthranilic acids. Incorporation of the unnatural precursors was quantified. Three fluorinated aurachin analogues were produced in sufficient quantity to enable their antibacterial activities to be assessed. Nature is a rich source of bioactive compounds, which have been optimized during evolution regarding their affinity to cellular targets.Stigmatella aurantiaca and later also found in other Stigmatella spp. as well as members of the actinomycete genera Rhodococcus and Streptomyces.[11]\u00a0Their structural relatedness to the known respiratory chain inhibitor 2\u2010heptyl\u20104\u2010hydroxyquinoline\u2010N\u2010oxide (HQNO) suggested early on that the aurachins might interfere with electron transport processes.6fb complex.1) and C (2).The aurachins are a large family of prenylated quinolone antibiotics, which were first discovered in cultures of the myxobacterium ptomyces.\u00a0Their st19F NMR spectroscopy.To explore this yet underexplored chemical space, we considered precursor\u2010directed biosynthesis as a viable option to introduce functional groups at these positions. Isotopic labelling studiesStigmatella erecta strain, which had been confirmed as an authentic aurachin producer.S. erecta in the presence of halogenated anthranilic acids, irrespective of the fluorine or chlorine substitution pattern. This may hint at interferences with tryptophan metabolism, as recently reported for mycobacteria.S. erecta, like many other myxobacteria, does not grow homogeneously in liquid media, but is known to form cellular aggregates.S. erecta, the amounts of biosynthesized aurachins were normalized to the production of myxothiazol A. This known secondary metabolite from S. erectaThe feeding studies were conducted in shaken Erlenmeyer flasks with a In accordance with previous studies, the addition of anthranilic acid was found to have a positive effect on aurachin production.19F NMR guided fractionation . The recovery of 6\u2010chloroaurachin D remained unsuccessful despite repeated attempts.In order to enable full NMR\u2010based structural characterization, feeding studies were repeated on multilitre scale to produce sufficient quantities of some aurachin analogues. Owing to the very low production titres that were achieved and a laborious chromatographic separation process Figure\u2005, only th1\u2009a (0.5\u2005mg) originated from a 10\u2010L culture of S. erecta grown in the presence of 3\u2010fluoroanthranilic acid. The [M+H]+ ion peak of 1\u2009a possesses a m/z value of 382.2546, corresponding to a molecular formula of C25H32NOF and 10 double bond equivalents. UV maxima at 241, 321 and 333\u2005nm as well as NMR data of the purified compound are almost consistent with those of 1,1H NMR spectrum of 1\u2009a features only three aromatic signals with a distinctive coupling pattern characteristic for a 3\u2010fluoroanthranilate\u2010derived moiety. COSY and HMBC experiments confirmed the presence of a farnesyl chain and a methyl group, which are connected to the assumed quinolone core of 1\u2009a at C\u20103 and C\u20102, respectively. Thus, 1\u2009a was identified as 8\u2010fluoroaurachin D.Compound 1\u2009b after multi\u2010step HPLC. The high\u2010resolution ESI\u2010MS spectrum confirmed the same molecular formula as 1\u2009a as well as similar UV maxima at 249, 321 and 331\u2005nm . A comparison of 1H NMR spectra corroborated that 1\u2009a and 1\u2009b are indeed structurally similar, but not identical. The differences pertained to the aromatic region. Following an analysis of their multiplicities and coupling constants, the aromatic proton signals of 1\u2009b could be assigned to a spin system for a 1,2,4\u2010trisubstituted benzene ring. Further interpretation of 1D and 2D NMR data then led to the conclusion that 1\u2009b represents 7\u2010fluoroaurachin D.The extract from the 4\u2010fluoroanthranilic acid culture yielded 0.3\u2005mg of compound 1\u2009c was purified from a culture that had been supplied with 5\u2010fluoroanthranilic acid, yielding 0.3\u2005mg of material. Again, high\u2010resolution ESI\u2010MS and UV data pointed to an aurachin D derivative bearing a single fluorine substituent . The substitution pattern with the halogen at C\u20106 position of the quinolone ring was deduced from the heteronuclear coupling observed in the 1H NMR spectrum .Compound 1\u2009a\u2013c are equipotent to 1 was grown in modified Zein medium . To facilitate the recovery of secreted aurachins, adsorber resin XAD7HP (2\u2009% w/v) was added prior to sterilization. The medium was inoculated with seed culture (10\u2009% v/v inoculum) and the fermentation was conducted in an incubator shaker at 30\u2009\u00b0C and 130\u2005rpm for 7\u2005days. Anthranilic acid or its halogenated derivatives were supplemented as filter\u2010sterilized aqueous solutions (33\u2005mg\u2009L\u22121) after inoculation of the medium. For extraction the adsorber resin was separated from the culture broth by filtration, washed with water and retained compounds eluted three times with acetone and methanol. The extract was dried and used for relative quantification or compound purification.Quantitative analysis: For relative quantification experiments, the dried extracts of 1.5\u2005L cultures were resuspended in methanol, filtered and supplied to LC\u2010ESI\u2010MS in positive mode. HPLC flow rate was 0.4\u2005mL\u2009min\u22121 on a Nucleodur RP 18 ec column and a gradient from 5 to 98\u2009% acetonitrile in water supplemented with 0.1\u2009% formic acid over 10\u2005min, followed by 5\u2005min at 98\u2009% acetonitrile. All analyses were carried out at a capillary voltage of 4.5\u2005kV, desolvation gas (N2) temperature of 220\u2009\u00b0C, with a dry gas (N2) flow rate of 12\u2005L\u2009min\u22121.Compound purification: Dried raw extracts were resuspended in 60\u2009% aqueous methanol (100\u2005mL) and extracted three times with dichloromethane (60\u2005mL). Aurachins were exclusively present in the dichloromethane fraction. The purification continued on a Shimadzu HPLC system (LC\u201020AD) equipped with a diode array detector (SPD\u2010M20A). The initial separation was carried out on a VarioPrep C18 Gravity column by two consecutive isocratic elutions with pure methanol and 90\u2009% aqueous methanol, respectively. The aqueous eluent was supplemented with 0.1\u2009% (v/v) trifluoroacetic acid and the flow rates were set to 4\u2005mL\u2009min\u22121. For 1\u2009c, an additional purification on a Nucleodur gravity column was performed (60\u2009% to 100\u2009% acetonitrile in water supplemented with 0.1\u2009% (v/v) trifluoroacetic acid within 10\u2005min, followed by 100\u2009% acetonitrile for additional 10\u2005min), at a flow rate of 2\u00a0mL\u2009min\u22121. NMR spectra were acquired at 300\u2005K on a Bruker AV 700 Avance III HD (CryoProbe) or on a Bruker AV 600 Avance III HD (CryoProbe). Compounds were dissolved in methanol\u20104d, which also served as internal standard to calibrate spectra to \u03b4H=3.31\u2005ppm and \u03b4C=49.0\u2005ppm. 19F NMR spectra were referenced to residual trifluoroacetic acid at \u03b4F=\u221276.5\u2005ppm.8\u2010Fluoroaurachin D (1\u2009a): 1H NMR : \u03b4=8.02 , 7.43 , 7.30 , 5.09 , 5.04 , 4.98 , 3.40 , 2.51 , 2.10 , 2.03 , 1.92 , 1.87 , 1.81 , 1.60 , 1.55 , 1.49\u2005ppm ; 13C NMR : \u03b4=177.8 (C\u20104), 153.3 , 150.4 (C\u20102), 136.1 (C\u20103\u2032), 135.9 (C\u20107\u2032), 132.0 (C\u201011\u2032), 130.1 (C\u20108a), 127.1 (C\u20104a), 125.3 (C\u20106\u2032), 125.3 (C\u201010\u2032), 123.9 , 123.5 (C\u20102\u2032), 121.7 (C\u20103), 122.0 , 116.7 , 40.8 (C\u20108\u2032), 40.7 (C\u20104\u2032), 27.8 (C\u20109\u2032), 27.2 (C\u20105\u2032), 25.8 (C\u201012\u2032), 24.8 (C\u20101\u2032), 18.0 (C\u20109), 17.6 (C\u201015\u2032), 16.3 (C\u201013\u2032), 16.2\u2005ppm (C\u201014\u2032); 19F NMR : \u03b4=133.5\u2005ppm ; HRMS (ESI): m/z calcd for C25H32FNO: 382.2541 &bk:[M+H]+; found: 382.2546.7\u2010Fluoroaurachin D (1\u2009b): 1H NMR : \u03b4=8.25 , 7.16 , 7.11 , 5.09 , 5.05 , 4.99 , 3.38 , 2.45 , 2.10 , 2.02 , 1.94 , 1.87 , 1.80 , 1.61 , 1.55 , 1.51\u2005ppm ; 13C NMR : \u03b4=177.9 (C\u20104), 165.8 , 150.0 (C\u20102), 141.4 (C\u20108a), 135.9 (C\u20107\u2032), 135.7 (C\u20103\u2032), 132.0 (C\u201011\u2032), 129.5 (C\u20105),125.4 (C\u201010\u2032), 125.2 (C\u20106\u2032), 123.9 (C\u20102\u2032), 121.9 (C\u20104a), 120.8 (C\u20103), 113.3 (C\u20106), 103.4 (C\u20108), 40.8 (C\u20104\u2032), 40.8 (C\u20108\u2032), 27.8 (C\u20109\u2032), 27.2 (C\u20105\u2032), 25.7 (C\u201012\u2032), 24.4 (C\u20101\u2032), 18.3 (C\u20109), 17.7 (C\u201015\u2032), 16.1 (C\u201013\u2032), 16.0\u2005ppm (C\u201014\u2032); 19F NMR : \u03b4=109.3\u2005ppm ; HRMS (ESI): m/z calcd for C25H32FNO: 382.2541 [M+H]+; found: 382.2549.6\u2010Fluoroaurachin D (1\u2009c): 1H NMR : \u03b4=7.84 , 7.56 , 7.45 , 5.09 , 5.05 , 4.99 , 3.40 , 2.47 , 2.09 , 2.02 , 1.94 , 1.88 , 1.81 , 1.61 , 1.55 , 1.51\u2005ppm ; 13C NMR : \u03b4=177.7 (C\u20104), 160.5 , 150.0 (C\u20102), 137.2 (C\u20108a), 136.1 (C\u20103\u2032), 135.9 (C\u20107\u2032), 132.1 (C\u201011\u2032), 125.4 (C\u20106\u2032), 125.3 (C\u201010\u2032), 126.2 (C\u20104a), 123.6 (C\u20102\u2032), 121.6 (C\u20108), 121.3 (C\u20107), 120.8 (C\u20103), 110.1 (C\u20105), 40.8 (C\u20104\u2032), 40.7 (C\u20108\u2032), 27.8 (C\u20109\u2032), 27.3 (C\u20105\u2032), 25.8 (C\u201012\u2032), 24.8 (C\u20101\u2032), 18.3 (C\u20109), 17.6 (C\u201015\u2032), 16.3 (C\u201013\u2032), 16.1\u2005ppm (C\u201014\u2032); 19F NMR : \u03b4=119.2\u2005ppm ; HRMS (ESI): m/z calcd for C25H32FNO: 382.2541 [M+H]+; found: 382.2541.Antibacterial activity screening: An agar diffusion assay was performed with the purified compounds 1 and 1\u2009a\u2013c to test their antimicrobial activities against Escherichia coli SG458, Pseudomonas aeruginosa SG137, Staphylococcus aureus (MRSA) 134/94 and Mycobacterium vaccae IMET 10670. For this purpose, 50\u2005\u03bcL compound test solution (500\u2005\u03bcg\u2009mL\u22121 in DMSO) was pipetted into agar plates with pre\u2010punched 9\u2005mm holes. Ciprofloxacin (5\u2005\u03bcg\u2009L\u22121) was used as positive control.The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "Scientific Reports 10.1038/srep40064, published online 06 January 2017Correction to: This Article contains errors in Figure\u00a0A) A total of 40 mice were subcutaneously inoculated with H460 and H460/MX20 cells (\u22485\u2009\u00d7\u2009106) in the right flank, respectively. (B) The changes in tumor volume and body weight over time following the implantation. Data points represented the mean\u2009\u00b1\u2009SD of tumor volumes and body weight from each group. n\u2009=\u200920. (C) Solid tumor formation rate of H460 and H460/MX20 cells (100%). (D) The selected cell xenografts were cut into about 5\u00a0mm\u2009\u00d7\u20095\u00a0mm and fixed with 10% neutral formalin. (E) ABCG2 expression analysis by immunohistochemistry in tumor tissues collected from H460 and H460/MX20 cell xenografts.\u201d\u201c(should read:A) A total of 40 mice were subcutaneously inoculated with H460 and H460/MX20 cells (\u2248 5\u2009\u00d7\u2009106) in the right flank, respectively. The changes in tumor volume and body weight over time following the implantation. Data points represented the mean\u2009\u00b1\u2009SD of tumor volumes and body weight from each group. n\u2009=\u200920. (D) Solid tumor formation rate of H460 and H460/MX20 cells (100%). (E) The selected cell xenografts were cut into about 5\u00a0mm\u2009\u00d7\u20095\u00a0mm and fixed with 10% neutral formalin. (F) ABCG2 expression analysis by immunohistochemistry in tumor tissues collected from H460 and H460/MX20 cell xenografts.\u201d\u201c(The correct Figure"} +{"text": "Slc17a9, P2x7r, and P2y2r was increased concomitant with an increase in extracellular ATP levels. Furthermore, compressive force decreased the osteoblast differentiation capacity of MC3T3\u2010E1 cells. shRNA knockdown of Slc17a9 in MC3T3\u2010E1 cells reduced levels of extracellular ATP and also led to increased osteoblast differentiation after the application of compressive force as assessed by qPCR analysis of osteoblast markers such as Runx2, Osterix, and alkaline phosphatase (ALP) as well as ALP activity. Consistent with these observations, knockdown of P2x7r or P2y2r by siRNA partially rescued the downregulation of osteoblast differentiation markers, caused by mechanical loading. In conclusion, our results demonstrate that VNUT is expressed in osteoblasts and that VNUT inhibits osteoblast differentiation in response to compressive force by mechanisms related to ATP release and P2X7R and/or P2Y2R activity.Osteoblasts release adenosine triphosphate (ATP) out of the cell following mechanical stress. Although it is well established that extracellular ATP affects bone metabolism via P2 receptors [such as purinergic receptor P2X7 (P2X7R) and purinergic receptor P2Y2 (P2Y2R)], the mechanism of ATP release from osteoblasts remains unknown. Recently, a vesicular nucleotide transporter that preserves ATP in vesicles has been identified. The purpose of this study was to elucidate the role of VNUT in osteoblast bone metabolism. mRNA and protein expression of VNUT were confirmed in mouse bone and in osteoblasts by quantitative real\u2010time PCR (qPCR) and immunohistochemistry. Next, when compressive force was applied to MC3T3\u2010E1 cells by centrifugation, the expression of In this study, we show that VNUT (SLC17A9), a vesicular nucleotide transporter, regulates osteoblast differentiation in response to mechanical loading. Mechanical loading promotes adenosine triphosphate (ATP) exocytosis via VNUT. Extracellular ATP subsequently inhibits osteoblast differentiation via activation of P2X7 and/or P2Y2 receptors. Thus, VNUT modulation of purinergic signaling contributes to the regulation of bone metabolism by mechanical loading. ALPalkaline phosphataseATPadenosine triphosphate P2X1Rpurinergic receptor P2X1P2X7Rpurinergic receptor P2X7P2Y2Rpurinergic receptor P2Y2qPCRquantitative real\u2010time PCRSLC17A9solute carrier family 17 member 9VNUTvesicular nucleotide transporterBone homeostasis is maintained by the balanced activity of bone\u2010forming osteoblasts and bone\u2010resorbing osteoclasts. Mechanical stress and loading are key factors for the regulation of bone homeostasis . OsteoblMechanical loading induces multiple cellular events in osteoblasts. A major effect of mechanical loading on osteoblasts is the efflux of adenosine triphosphate (ATP) into the extracellular space , 13, 14.Solute carrier family 17 member 9 (SLC17A9) was recently identified as a vesicular nucleotide transporter (VNUT), essential for ATP secretion from adrenal PC12 cells . ATP canHere, we report that VNUT is expressed in osteoblasts where it plays a role in ATP secretion as well as osteoblast differentiation.\u22121 penicillin\u2013streptomycin . Osteoblast differentiation was induced by culturing cells in an osteogenic medium containing 50\u00a0\u03bcg\u00b7mL\u22121 ascorbic acid and 10\u00a0mm \u03b2\u2010glycerophosphate for 7\u00a0days. Cells were treated with 1, or 10\u00a0\u03bcm clodronate (Sigma\u2010Aldrich). Ionomycin treatments were performed at 1\u00a0\u03bcm. Brefeldin A was used at 10\u00a0\u03bcm. HEK293T cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS (Biosera Nuaill\u00e9) and 100\u00a0U\u00b7mL\u22121 penicillin\u2013streptomycin .MC3T3\u2010E1 cells, a mouse calvarial osteoblast cell line, were obtained from Riken Bio Resource Center . MC3T3\u2010E1 cells were maintained in \u03b1\u2010minimal essential medium supplemented with 10% FBS and 100\u00a0U\u00b7mL5 cells per well in 12\u2010well plates. Medium was changed into 4\u2010(2\u2010hydroxyethyl)\u20101\u2010piperazineethanesulfonic acid (HEPES)\u2010buffered DMEM without bicarbonate , supplemented with 18\u00a0\u00b5m of hydrochloric acid to stabilize pH and then cells placed in an incubator at 37\u00a0\u00b0C for 1\u00a0h. The plates were subsequently centrifuged at 4.4\u00a0\u00d7\u00a010\u22122\u00a0N\u00b7cm\u22122 (5.0\u00a0g\u00b7cm\u22122) or 8.8\u00a0\u00d7\u00a010\u22122\u00a0N\u00b7cm\u22122 (9.0\u00a0g\u00b7cm\u22122) using a PlateSpin II centrifuge in the incubator at 37\u00a0\u00b0C for 12\u00a0h. Compressive force by weights was also previously described . Boxed areas in the left panel are shown as magnified images in the right panel. Scale bars indicate 20 \u00b5m and 100 \u00b5m respectively.Click here for additional data file.Fig. S2. Immunocytochemistry with VNUT antibodies. (A) MC3T3\u2010E1 cells are immunostained with Rabbit VNUT antibody preabsorbed with VNUT blocking antigen peptide (Control) or VNUT antibody (Rabbit). (B) MC3T3\u2010E1 cells stably expressing control scrambled shRNA or shRNA against murine Slc17a9 were stained with VNUT antibody (Rabbit), rhodamine phalloidin, or DAPI. (C) MC3T3\u2010E1 cells were transfected with V5\u2010tagged VNUT and immunostained with VNUT antibody (Rabbit or Guinea pig), and anti\u2010V5 antibody and DAPI. Scale bars indicate 100 \u00b5m, scrambled shRNA, Scr; shRNA against murine Slc17a9, Sh.Click here for additional data file.Fig. S3. Knock down of VNUT does not alter extracellular ATP levels in the absence of mechanical force. Extracellular ATP levels from MC3T3\u2010E1 cells stably expressing control scrambled shRNA or shRNA against Slc17a9. Scrambled shRNA, Scr; shRNA against murine Slc17a9, sh. Data are expressed as the mean\u00a0\u00b1\u00a0SD (n\u00a0=\u00a03). Statistical analysis was performed with unpaired t\u2010test. *P\u00a0< 0.05 or **P\u00a0<\u00a00.01 versus control.Click here for additional data file.Fig. S4. Inhibition of Vnut function does not affect cell proliferation of MC3T3\u2010E1 cells. MC3T3\u2010E1 cells were plated into 96\u2010well plates and incubated with 0, 1.0, or 10 \u03bcM clodronate. Cell proliferation was assessed on day 1, 2, or 3 using a Cell Counting Kit\u20108 (DOJINDO).Click here for additional data file."} +{"text": "IVX5}\u2212 synthon to a pocket\u2010based ligand to provide supramolecular design using halogen\u22c5\u22c5\u22c5halogen interactions within an FeIII system that has the potential to undergo spin crossover (SCO). By removing the solvent from the crystal lattice, we \u201cswitch on\u201d halogen\u22c5\u22c5\u22c5halogen interactions between neighboring molecules, providing a supramolecular cooperative pathway for SCO. Furthermore, changes to the halogen\u2010based interaction allow us to modify the temperature and nature of the SCO event.We have added the {Re Inorganic synthon controls SCO behavior: Incorporating an {ReIVX5}\u2212 synthon into an FeIII system provides a highly directional supramolecular design using halogen\u22c5\u22c5\u22c5halogen interactions. Removal of solvent from the crystal lattice \u201cswitches on\u201d these interactions, providing a long\u2010range cooperative pathway ideal for an SCO event. In practice we found that the transition metal preferentially occupies pocket 2, giving a mer\u2010coordinated system with the general formula [ReIVX5(\u03bc\u2010Rpch)Fe(R\u2019\u2010Im)3] where H2Rpch has the formula (E)\u2010N\u2019\u2010(2\u2010hydroxy\u20103\u2010R\u2010benzylidene)pyrazine\u20102\u2010carbohydrazide, and R\u2019\u2010Im has the formula 1\u2010R\u2019\u2010imidazole \u2010(8) (see ESI), showed that the SCO properties of the FeIII center could be readily modified. Crystal structures are labelled with the general name \u201c(Compound)w/d_TEMP\u201d, where w or d denotes \u201cwet\u201d (solvated) or \u201cdry\u201d (desolvated) crystals, and TEMP is the temperature of the structural determination.Whilst the use of the {ReX1)w_100 shows that the complex crystallizes in the space group C2/c with Z=8. Complex (1) was chosen as a representative of the series with structural details of complexes (1)\u2013(4) including halogen bond parameters in the ESI. The asymmetric unit contains a [ReBr5(\u03bc\u2010MeOpch)Fe(Me\u2010Im)3] complex with the {ReIVBr5} synthon bound to a nitrogen atom of the pyrazine ring of the deprotonated MeOpch ligand. The central FeIII ion is coordinated to the ligand through an azine nitrogen (N1), a carbonyl oxygen (O3), and a hydroxy oxygen (O1), whilst three additional Me\u2010Im co\u2010ligands complete the coordination sphere through nitrogen atoms N5, N9, and N7. In addition to this there were two interstitial MeCN molecules within the lattice connected through nonclassical hydrogen bonds. The shortest intermolecular Br\u22c5\u22c5\u22c5Br distance was 4.663\u2005\u00c5 and is therefore too long to be considered a halogen bond, and is unlikely to contribute to information exchange.4O2 coordination sphere of the FeIII ion is often observed in SCO active compounds,1)w_280. FeIII\u2212N bond lengths are characteristic of the spin state (LS\u22481.95\u2005\u00c5 and HS\u22482.15\u2005\u00c5)1) using a SQUID magnetometer revealed an SCO active system with an incomplete transition occurring between 220\u2005K and 90\u2005K, from fully HS to only 22\u2009% of molecules in the HS electronic configuration with T1/2=155\u2005K d_100 crystallized in the same space group, C2/c, with the same neutral metal complex in the asymmetric unit. However, the average Fe\u2212N bond length (1.982\u2005\u00c5) indicated the complex was now at least partially in the LS state; it was also observed that the sample had desolvated (between measurements) with loss of the interstitial MeCN molecules whilst retaining crystallinity. Crystallographic data were also collected at 140, 180, 230 and 280\u2005K and by plotting the adjusted change in unit cell volume (see ESI for details) against the temperature the SCO event clearly correlates with the changes in the crystal structure , however the loss of the solvent leads to reorientation of molecules in the crystal resulting in a significant shortening of the Br\u22c5\u22c5\u22c5Br distances, \u201cswitching on\u201d a type I halogen bond .A more detailed investigation revealed that, by removing the lattice solvent and therefore the hydrogen bonding, halogen\u22c5\u22c5\u22c5halogen interactions become the dominant supramolecular effect Figure\u2005. In the 2)\u2013(4) were then synthesized to characterize the importance of these halogen\u22c5\u22c5\u22c5halogen interactions. The modifications are summarized in Table\u20052). Complex (2)w_100 also crystallized in the space group C2/c with the neutral metal complex now [ReCl5(\u03bc\u2010MeOpch)Fe(Me\u2010Im)3] together with two MeCN molecules in the asymmetric unit. Analysis of the Fe\u2212N bond lengths (average: 2.111\u2005\u00c5) shows the iron(III) center to be in the HS state at 100\u2005K. In this solvated sample there is again no appreciable halogen\u22c5\u22c5\u22c5halogen interaction, with the shortest Cl\u22c5\u22c5\u22c5Cl distance at 4.539\u2005\u00c5. Despite the apparent HS structure, magnetic investigation revealed that at 100\u2005K the FeIII is mostly LS (74\u2009% LS) with the halogen bond \u201cswitching on\u201d upon loss of solvent. Since this sample lost crystal quality during desolvation, the exact nature of the halogen\u22c5\u22c5\u22c5halogen interaction could not be clearly defined. Previous work has ordered the strength of halogen bonds as F < Cl
\u20090.95 and (700\u2013800\u00a0\u00b0C) \u03b7\u2009=\u20090 for the (B) samples and \u03b7\u2009=\u20090.95\u20130.8 for the (A) samples in the specified temperature ranges. In the temperature interval 200\u2013300\u00a0\u00b0C, the equality \u03b7\u2009=\u20090 is determined by the formation of a non-magnetic nfm-B2 phase. Although the \u03b1l\u02b9 (001) layer is located between the Rh and Fe (001) layers and grows epitaxial on the surface of Fe (001) in the (400\u2013600\u00a0\u00b0C) interval it has a reversible \u03b1l\u02b9\u2009\u2194\u2009\u03b1\u02ba transition with a low residual magnetization samples. We hypothesize that the \u03b1\u02b9 phase has two B2-ordered polymorphic modifications having similar lattice parameters, but different magnetic properties. In the first modification the \u03b1l\u02b9 phase has a low magnetization about 825\u00a0emu/cm3, which is defined only by the Fe atoms with the Rh atoms not contributing to the magnetization, and the \u03b1l\u02b9 undergoes a complete reversible \u03b1l\u02b9\u2009\u2194\u2009\u03b1\u02ba transition samples, the Fe atoms supposedly polarize the Rh and the Rh atoms make the contribution to the saturation magnetization 1,220\u00a0emu/cm3 and in the \u03b1h\u02b9 phase the \u03b1h\u02b9\u2009\u2194\u2009\u03b1\u02ba transition is completely suppressed and with an increase in annealing temperature the phase sequence 52Rh/48Fe\u2009\u2192\u2009(~\u2009100\u00a0\u00b0C) nfm-B2\u2009\u2192\u2009(300\u00a0\u00b0C) \u03b1l\u02b9\u2009\u2192\u2009(600\u00a0\u00b0C) \u03b1l\u02b9 or \u03b1h\u02b9 is formed as shown in Fig.\u00a0In experiments, all 52Rh/48Fe001) samples, after annealing each in a temperature range of 100\u2013800\u00a0\u00b0C, were subjected to an investigation of the \u03b1\u02b9\u2009\u2194\u2009\u03b1\u02ba transition, consisting of measurements of the magnetic moment m layers on the MgO(001) surface. The K4(Ta)/K40 and MS/MS0 dependencies (where for 45Rh/55Fe(001) samples K40\u2009=\u20092.5\u00a0\u00d7\u00a0105\u00a0erg\u00a0cm3 and MS0\u2009=\u2009875\u00a0emu/cm3, see \u201cl\u02b9(001) with\u00a0low magnetization (MS/MS0\u2009~\u20091.0) grows into the high quality epitaxial phase \u03b1h\u02b9(001) with\u00a0high magnetization (MS/MS0\u2009~\u20091.47) above 450\u00a0\u00b0C. The high quality of the chemical ordering of the \u03b1h\u02b9(001) films after annealing at 500\u00a0\u00b0C and 800\u00a0\u00b0C supports the order parameter S\u2009=\u20090.96\u2009\u00b1\u20090.02, which is more than the S\u2009=\u20090.90\u2009\u00b1\u20090.02 for \u03b1l\u02b9 in 52Rh/48Fe films. This result is unexpected, since the \u03b1l\u02b9 exists in a narrow composition range of nearly equiatomic concentration and must have a more complete B2 order than the Fe-rich \u03b1h\u02b9 phase. The \u03b1h\u02b9(001) and \u03b1l\u02b9(001) films have the same orientation relationship with the substrate MgO(001) /K40 dependence and the AFM-FM transition has a relatively low residual magnetization \u03b1h\u02b9. Finally, we proved that even a slight Fe doping of \u03b1l\u02b9 causes a chemical reaction between Fe and \u03b1l\u02b9 and the start of the synthesis of the \u03b1h\u02b9 phase. This suggests that the high magnetization \u03b1h\u02b9 phase occurring in the 52Rh/48Fe(001) bilayer (B samples) also has more Fe content than \u03b1l\u02b9 and explains the compositional heterogeneity arising as a result of the nonequilibrium reaction processes. Thus, we show the phase evolution 45Rh/55Fe\u2009\u2192\u2009(~\u2009100\u00a0\u00b0C) nfm-B2\u2009\u2192\u2009(300\u00a0\u00b0C) \u03b1l\u02b9\u2009\u2192\u2009(450\u00a0\u00b0C) \u03b1h\u02b9 is induced by the solid-state-reaction method and the final reaction product is a highly B2-ordered phase \u03b1h\u02b9, which has a high magnetization of 1,270\u00a0emu/cm3 and in which the reversible \u03b1h\u02b9\u2009\u2194\u2009\u03b1\u02ba transition is completely suppressed.The schematic diagram in Fig.\u00a0Fig.\u00a0h\u02b900 layers ol\u02b9 phases during annealing up to 500\u00a0\u00b0C in the 68Rh/32Fe, 52Rh/48Fe interface layer in the Rh/\u03b1l\u02b9(001) bilayer system surface originating from the solid-state reaction Rh\u2009+\u2009\u03b1l\u02b9\u2009\u2192\u2009(500\u00a0\u00b0C) \u03b3. As shown in Fig.\u00a0S(Ta)/MS0, and the K4(Ta)/K40 dependence presented in Fig.\u00a0l\u02b9 above 300\u00a0\u00b0C and the paramagnetic \u03b3 phases above 500\u00a0\u00b0C. As follows from Fig.\u00a0l\u02b9 phase exists and undergoes a reversible AFM-FM transition (\u03b7\u2009~\u20090.9) in the 300\u2013500\u00a0\u00b0C temperature range nfm-B2\u2009\u2192\u2009(300\u00a0\u00b0C) \u03b1l\u02b9\u2009\u2192\u2009(500\u00a0\u00b0C)\u03b3 during thermal annealing up to 800\u00a0\u00b0C and the final reaction product contains only an epitaxial \u03b3(001) layer on the MgO(001) surface.Figure\u00a08Fe Fig.\u00a0a and 55Rl\u02b9\u2009\u2192\u2009\u03b1\u02ba transition, the synthesis kinetics of the nfm-B2 layer during isothermal aging at 110\u00a0\u00b0C was investigated. As stated above, deposition of Rh on Fe(001) at room temperature at any ratio of thicknesses between Rh and Fe leads to the formation of a thin interfacial nanocrystalline nfm-B2 layer. However, slight heating to 100\u00a0\u00b0C during sputtering cannot be excluded, therefore, aging of as-deposited samples was investigated at 110\u00a0\u00b0C. XRD data from the as-deposited Rh/Fe(001) bilayer and after aging up to 360\u00a0h shows a small broadened (001) superlattice and the fundamental (002) peaks of the nfm-B2 phase which did not change from aging time to 360\u00a0h layer, which entered into a reaction with Rh at 110\u00a0\u00b0C, as a function of aging time. There is a large experimental error in the thickness dFe measurements and therefore the dependence dFe(t) is impossible to describe by an unambiguous kinetic equation, which suggests a possible growth mechanism. As shown in Fig.\u00a0Fe after aging for 360\u00a0h at 110\u00a0\u00b0C is\u2009~\u20098\u00a0nm, which corresponds to the thickness dnfm-B2\u2009~\u200917\u00a0nm of the nfm-B2 layer interface at the temperature of the \u03b10\u00a0h Fig.\u00a0a. This m\u00a0nm Fig.\u00a0b. Figurel\u02b9 around 300\u00a0\u00b0C, we investigated the evolution of the temperature dependencies of the magnetization and the thickness of the interfacial hB2-FeRh layer in the 52Rh/B2-FeRh/48Fe(001) trilayer after annealing at 280\u00a0\u00b0C, 300\u00a0\u00b0C and 350\u00a0\u00b0C. Figure\u00a0nfm-B2 interlayer in the 48Rh/nfm-B2/52Fe(001) trilayer increases to a thickness around dnfm-B2 ~ 30\u00a0nm after annealing at 280\u00a0\u00b0C. \u2009~\u200920\u00a0nm of the \u03b1l\u02b9 layer in the B2-FeRh interlayer. This clearly proves that after annealing at 300\u00a0\u00b0C the B2-FeRh interlayer contains a mixture of 50% nfm-B2 and 50% \u03b1l\u02b9 phases /K40 dependence /K40 \u2009<\u2009150\u00a0\u00b0C49, Tin(Ni/Al)\u2009~\u2009180\u00a0\u00b0C50, Tin(Cd/Au)\u2009=\u200967\u00a0\u00b0C51 in Ti/Ni, Ni/Al, Cd/Au bilayers, respectively. These temperatures are close to or coincide with the reverse\u00a0martensitic transformation starting temperatures\u00a0As(B2-TiNi)\u2009~\u2009100\u00a0\u00b0C, As(B2-NiAl)\u2009~\u2009180\u00a0\u00b0C, As(B2-CdAu)\u2009=\u200967\u00a0\u00b0C. The reversible \u03b1l\u02b9\u2009\u2194\u2009\u03b1\u02ba transition in B2-FeRh possesses all the characteristics of a martensitic transformation , because it can pass at high speeds53, has isotropic volume changes at the transition1, can be induced by the application of stress40 and magnetic field10, has martensitic instabilities54 and the \u03b1l\u02b9 and \u03b1\u02ba lattices have a cube-on-cube orientation relationship. According to the phase diagram, the transition \u03b1l\u02b9\u2009\u2194\u2009\u03b1\u02ba has a minimum temperature Tk\u2009~\u2009100\u00a0\u00b0C among other structural transformations in the Fe-Rh system. From the above, we have concluded that the initiation temperature of the reaction Tin(Rh/Fe) in the Rh/Fe bilayer coincides with the martensitic-like transition temperature TK\u2009~\u2009100\u00a0\u00b0C in B2-FeRh. The coincidence of the starting temperature of the reaction between Fe and Rh and the temperature of the magnetostructural AFM-FM transition suggests common chemical mechanisms behind both phenomena, but that connection remains to be confirmed by additional experimentation.It is well established that ordered B2 alloys, such as NiTi, AuCd, NiAl have reversible low-temperature martensitic transformations, in which the high-temperature austenite B2-phase develops into a low-temperature martensitic phase through a complex process of the55 and found in the experimental study36. Competition between the \u03b1l\u02b9 and \u03b1\u02ba phases during the partial crystallization suppresses grain growth and stabilizes the nano-grained B2 structures in an amorphous matrix. The formation of an amorphous phase is a quite common phenomenon in the initial stage of solid-state reactions in bilayers and multilayers, although the nature of this phenomenon is still a subject of dispute58. It is interesting to note the general features of the initial stage of the synthesis of B2 phases in Ti/Ni and Rh/Fe thin films. In Ti/Ni multilayers the amorphous phase starts near the martensitic transition temperature (~\u2009100\u00a0\u00b0C), which turns into B2-NiTi60 at annealing temperatures above 350\u00a0\u00b0C. Similar to Ti/Ni, the reaction in the Rh/Fe(001) bilayer starts at\u2009~\u2009100\u00a0\u00b0C with the formation of the interfacial amorphous phase, which partially crystallizes. From this point the as-deposited films consist of nanocrystalline B2 grains dispersed in the amorphous matrix. This strongly suggests that both amorphous phases are amorphous martensite, which may be a universal phenomena of the solid-state synthesis of martensitic phases61. This scenario is different from the synthesis of B2-NiAl and B2-AuCd, which begin to form in Al/Ni50 and Cd/Au51 bilayers at martensitic transformation temperatures without the formation of an intermediate amorphous martensite.As mentioned above, the formation of compounds at the Rh/Fe interface starts in the temperature range 100\u2013300\u00a0\u00b0C from the synthesis of the non-ferromagnetic nfm-B2 phase. Since this phase has only (001) and (002) B2-FeRh reflections, this means that nfm-B2 is either a martensitic-like antiferromagnetic \u03b1\u02ba-phase or a non-ferromagnetic martensitic variant which is stabilized by strains resulting from the non-equilibrium synthesis of the nfm-B2 phase. B2-FeRh, similar to other B2 phases of NiTi, AuCd, NiAl alloys, experiences premartensitic instabilities with the subsequent formation of the different\u00a0structural variants of martensite. Our hypothesis is the amorphous phase appears above 100\u00a0\u00b0C at the Rh/Fe interface, which is then transformed into a\u00a0nanocrystalline state containing B2 nanograins of martensite variants with lattice parameters close to \u03b1\u02ba martensite.\u00a0This is consistent with the possible existence of various structural phases of FeRh, predicted by ab initio calculationsin(\u03b1h\u02b9)\u2009=\u2009\u2009~\u2009450\u00a0\u00b0C and Tin(\u03b3)\u2009=\u2009\u2009~\u2009500\u00a0\u00b0C of the \u03b1h\u02b9 and \u03b3 phases coincide with the phase transition temperatures in the Fe-rich and Rh-rich regions of the Fe\u2013Rh system, respectively. Such an approach is justified by us for the well-studied Fe\u2013Ni system62 and made it possible to predict phase transformations in other binary metallic systems48. Therefore, further study of solid-state reactions in Rh/Fe films, depending on the composition, will make it possible to specify the low-temperature part of the Fe\u2013Rh phase diagram, which still remains unknown63.Analysis of the general thermodynamic characteristics and features of B2-FeRh and B2-NiTi suggests the possibility of a fabrication of the B2-FeRh compound by self-propagating synthesis samples and the \u03b1l\u02b9 reacted with Ph and formed the non-ferromagnetic \u03b3 phase in the 68Rh/32Fe(001)samples. Magnetic analysis has revealed that only the \u03b1l\u02b9 undergoes the complete reversible \u03b1l\u02b9\u2009\u2194\u2009\u03b1\u02ba transition and there\u2019s no transition in the \u03b1h\u02b9 samples. Thus, our work not only provides an idea of how phase sequences start and develop depending on the composition of the Fe/Rh bilayers, but also suggests the interesting possibility that a similar chemical mechanism may be at play behind the low-temperature reaction of Fe and Rh and the AFM-FM transition in B2-FeRh.In conclusion, we have uncovered that regardless of the Rh and Fe thicknesses the thin nonmagnetic nanocrystalline B2-FeRh layer starts to form at\u2009~\u2009100\u00a0\u00b0C and grows up to 300\u00a0\u00b0C on the Rh/Fe interface. Above 300\u00a0\u00b0C the nonmagnetic phase is converted into a low-ferromagnetic B2 \u03b14, which coincided with the value of bulk iron K1\u2009=\u20094.9\u2009\u00d7\u2009105\u00a0erg/cm3. The constant K4 in the (001) plane was determined by a torque magnetometer in a magnetic field H\u2009=\u200912 kOe. The torque curve L\u01c0\u01c0(\u03c6) in the (001) plane was determined according to the equation 2L\u01c0\u01c0(\u03c6)\u2009=\u2009K4Vsin4\u03c6\u2009+\u20092 Ku VSin(2\u03c6\u2009+\u2009\u03b3), in which, in addition to the dominant 4\u03c6-term, there is a minor 2\u03c6-term Ku term due to the surface roughness of the MgO(001) substrate and a slight misorientation of the Fe(001) grains. In the equation Ku is uniaxial anisotropy constant, V is the volume of the film, \u03c6 is the angle between the easy axis of fourfold anisotropy and the magnetization MS, \u03b3 is the angle between the easy axis of fourfold anisotropy and the axis of the uniaxial anisotropy. The K4V value was calculated from the torque curve L\u01c0\u01c0(\u03c6) at the maximum of the 4\u03c6-term: 2Lmax\u2009=\u2009K4V. High quality epitaxial Fe(001)/MgO(001) films also can be obtained by various other methods as reported in the literature.At first the epitaxial Fe(001)/MgO(001) films were grown on single-crystal MgO(001) substrates\u00a0by a\u00a0thermal\u00a0evaporation\u00a0method in a vacuum chamber at a pressure of 10\u20136\u00a0mbar. To obtain high-quality Fe(001) films, the substrates was previously outgassed at 300\u00a0\u00b0C for 1\u00a0h and the Fe layers were deposited at 250\u00a0\u00b0C. Epitaxial Fe (001) films had the orientation ratio Fe(001),[100]||MgO(001),[110] with the MgO(001) substrate and the magnetocrystalline\u00a0anisotropy constant K\u20136\u00a0mbar, and a working pressure of\u2009~\u20091 mTorr Ar was used during sputtering. To prevent a reaction between Rh and Fe, the Rh layer was deposited at room temperature. Under such deposition conditions the polycrystalline Rh layer was formed on the Fe(001) surface. Three different types of samples were prepared for the experiments, namely Rh/Fe (001) bilayers with Rh-rich, approximately 1Fe:1Rh and Fe-rich atomic ratios, each with a total thickness about 300\u00a0nm. The elemental chemical composition determined by energy dispersion X-ray (EDX) analysis showed sample compositions of Rh68Fe32, Rh52Fe48 and Rh45Fe55, respectively. The saturation magnetization MS0 and the magnetic fourfold anisotropy constants K40 were determined for the total volume of the 52Rh/48Fe, 45Rh/55Fe and 68Rh/32Fe bilayers, which turned out to be MS0\u2009=\u2009825 emu/cm3, K40\u2009=\u20092.2\u00a0\u00d7\u00a0105\u00a0erg/cm3 for the 52Rh/48Fe bilayers, MS0\u2009=\u2009875 emu/cm3, K40\u2009=\u20092.4\u00a0\u00d7\u00a0105\u00a0erg/cm3 for the 45Rh/55Fe bilayers and MS0\u2009=\u2009450 emu/cm3, K40\u2009=\u20091.25\u00a0\u00d7\u00a0105\u00a0erg/cm3 for the 68Rh/32Fe bilayers.The starting Rh/Fe(001) bilayers were obtained by the evaporation of the Rh layers on Fe(001)/MgO(001) samples using dc sputtering in a magnetron sputtering system. The base pressure of the chamber was less than\u2009~\u20091\u00a0\u00d7\u00a010S and the coercivity HC were measured with a vibration magnetometer in magnetic fields up to 22 kOe. All saturation magnetization measurements were monitored using the torque method64.The reversible AFM-FM phase transitions were checked using a superconducting quantum interference device (SQUID) magnetometer. Magnetic fields of H\u2009=\u20091.0 kOe were applied along the in-plane [100] MgO direction, which coincides with the easy axis of the Rh/Fe (001) bilayers, at all measurements in the 77\u2013400\u00a0K temperature interval. The saturation magnetization M001/I002)1/2/1.07, where I001 and I002 are experimental integrated intensities of superstructural (001) and fundamental (002) reflections65.The formed phases were identified with a DRON-4-07 diffractometer (CuKa radiation). The epitaxial relationships between MgO(001) and the B2, \u03b3 layers that formed in the reaction products were X-ray studied with a PANalytikal X'Pert PRO diffractometer with a PIXctl detector. CuKa radiation monochromatized by a secondary graphite monochromator was used in the instrument. The order parameter for the synthesized samples at temperatures in the 600\u2013800\u00a0\u00b0C range was estimated using the equation S\u2009=\u2009 at 40\u00a0kV. In order to protect the surface of interest from milling by the Ga\u2009+\u2009ion beam during sample preparation, a Ge layer was deposited onto the Fe-Rh film before cross-sectional sample preparation by FIB. TEM studies were carried out using a Hitachi HT7700 TEM equipped with a STEM system and a Bruker Nano XFlash 6T/60 energy dispersive X-ray (EDX) spectrometer. The imaging and EDX spectroscopy line scans and mapping were carried out in STEM mode with an electron probe of diameter\u2009~\u200930\u00a0nm.\u22126\u00a0Torr for 1\u00a0h. To characterize the phase transformations the crystal structure, the magnetic moments m0(Ta) (where m0(300\u00a0K)\u2009=\u2009MS0V), the magnetic fourfold anisotropy constants K4(Ta) and the degree of the \u03b1\u02b9\u2009\u2192\u2009\u03b1\u02ba transition \u03b7(Ta) were determined for all bilayers after annealing at each temperature Ta. In order to fully characterize the phase transformations, cross-sectional SEM images and an elemental analysis of the phases of several samples using EDX were carried out.The starting 52Rh/48Fe, 45Rh/55Fe and 68Rh/32Fe bilayers were annealed at temperatures ranging from 50 to 800\u00a0\u00b0C in increments of 50\u00a0\u00b0C. The samples were held at each temperature at a pressure of 10a) of the FM-AFM phase transition was determined for all 52Rh/48Fe, 45Rh/55Fe and68Rh/32Fe samples after annealing at each temperature in the range of 100\u2013800\u00a0\u00b0C . The magnetic moments m0(Ta) were measured by the torque method64, after annealing at temperature Ta and cooling in liquid nitrogen mN(Ta), to find the value\u00a0of the degree \u03b7(Ta). The magnetic moment m(Ta) of the synthesized \u03b1l\u02b9\u2014FeRh layer in the Rh/\u03b1l\u02b9\u2014FeRh/Fe(001) trilayer after annealing at temperature Ta is equal to the difference m(Ta)\u2009=\u2009m0(Ta)\u2014K4(Ta)m0/K40 of the magnetic moments of the m0(Ta) film and the K4(Ta)m0/K40 unreacted layer of the Fe(001) layer, where K4(Ta)m0/K40 is the magnetic moments of the unreacted Fe(001) layer. After placing the sample in liquid nitrogen, only the ferromagnetic \u03b1l\u02b9\u2014FeRh phase in the Rh/\u03b1l\u02b9\u2014FeRh/Fe(001) trilayer is subjected to the transition into the antiferromagnetic \u03b1\u02ba phase and the magnetic moment m(Ta) is reduced to mN(Ta). The quantity \u03b7\u2009=\u20091\u00a0\u2212\u00a0mN(Ta)/m(Ta) is a quantitative characteristic of the degree of the \u03b1l\u02b9\u2009\u2192\u2009\u03b1\u02ba transition, where \u03b7\u2009=\u20090 and \u03b7\u2009=\u20091 mean the absence of and the complete FM\u2009\u2192\u2009AFM transition , respectively.In experiments, the degree \u03b7(TSupplementary file1 (DOCX 4798 kb)"} +{"text": "The crystal packing features N\u2014H\u22efN hydrogen bonds, which generate [101] chains.The title structure consists of [Ni(CN) 10H10N3)2[Ni(CN)4], the dihedral angle between the pyridine rings in the cation is 1.92\u2005(13)\u00b0 and the complete anion is generated by a crystallographic centre of symmetry. An intra\u00admolecular N\u2014H\u22efN hydrogen bond occurs in the cation, which closes an S(6) ring. In the crystal, the components are linked by N\u2014H\u22efN and weak C\u2014H\u22efN hydrogen bonds, which generate chains propagating in the [101] direction. Weak aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions are also observed. A Hirshfeld surface analysis and two-dimensional fingerprint plots indicate that the most important contact types in the crystal packing are N\u22efH/H\u22efN, C\u22efH/H\u22efC and H\u22efH with contributions of 37.2, 28.3 and 21.9%, respectively.In the title mol\u00adecular salt, (C The C\u2014Non Fig.\u00a01. The cyanide ligands, exhibiting a square-planar geometry. The bond lengths and angles in the anion are in good agreement with those found in other [Ni(CN)4]2\u2212 salts \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01) link (Table\u00a01H) forms a strong N3\u2014H3A\u22efN2 hydrogen bond with a cyano group acceptor and the H3A\u22efN2 distance is 2.0\u2005\u00c5. Fig.\u00a03S(6) centroids and pyridyl groups [centroid\u2013centroid distance of 3.487\u2005(2)\u2005\u00c5].Fig.\u00a03k Table\u00a01. The secet al., 2007CrystalExplorer 17.5 . Areas on the Hirshfeld surface with high curvedness can be related to the planar packing arrangement of the cations. The most abundant inter\u00admolecular inter\u00adactions in the crystal packing \u00b7[CuCl4] . The mixture was sealed in a Teflon-lined autoclave and held at 423\u2005K for 3\u2005d, and then cooled to room temperature at a rate of 10\u2005K per hour (yield 27%). Pale-yellow plates of (I)The title compound was synthesized solvothermally under autogenous pressure using a mixture of iron(II) sulfate hepta\u00adhydrate , 2,2\u2032-di\u00adpyridyl\u00adamine and potassium tetra\u00adcyano\u00adnickelate(II) in mixed solvents of water/ethanol (3:1 Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698902001419X/hb7948sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902001419X/hb7948Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902001419X/hb7948sup3.tifFigure S1 Hirshfeld surface of (C10H10N3)2[Ni(CN)4] mapped with shape index (a) and curvedness (b). DOI: 2040378CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(6) graph-set motif. Inter\u00admolecular \u03c0\u2013\u03c0 stacking and C\u2014O\u22ef\u03c0 inter\u00adactions dominate the crystal packing.A short [2.592\u2005(3)\u2005\u00c5] intra\u00admolecular N\u2014H\u22efO hydrogen bond leads to an 13H12N2O2, crystallizes in the ortho\u00adrhom\u00adbic space group Pna21 with two very similar mol\u00adecules in the asymmetric unit. An intra\u00admolecular N\u2014H\u22efO hydrogen bond leads to an S(6) graph-set motif in each of the mol\u00adecules. Inter\u00admolecular \u03c0\u2013\u03c0 stacking and C=O\u22ef\u03c0 inter\u00adactions involving the aldehyde O atoms link mol\u00adecules into stacks parallel to [100]. A Hirshfeld surface analysis indicates that the most important contributions to the crystal packing stem from H\u22efH (49.4%) and H\u22efO/O\u22efH (21.5%) inter\u00adactions. Energy framework calculations reveal a significant contribution of dispersion energy. The crystal studied was refined as a two-component inversion twin.At 100\u2005K, the title compound, C Two major aspects contribute to the inter\u00adest in modified structural analogues of quinazoline alkaloids. On the one hand, they are attractive targets for the development of methods in organic synthesis; reactions sufficiently general to target a wide range of derivatives of a given lead structure should be easy to carry out and warrant high yields. On the other hand, substituted quinazolines allow the study of structure\u2013property relationships with respect to their biological activities was first isolated from the plant Mackinlaya subulata Philipson may be the reason for this selectivity towards primary amines.The title compound, 6-formyl-7,8,9,11-tetra\u00adhydro-51) Fig.\u00a01, does re1) Fig.\u00a01, 2016 \u25b8,1H NMR data and quantum-chemical calculations, Zhurakulov et al. (20161) in the solid state, we studied its mol\u00adecular and crystal structure. We also report the analysis of the Hirshfeld surface and the energy framework of crystalline (1).Based on al. 2016 confirmeA and B . In contrast to the quinazolinone moiety, the alkyl ring is not planar. The maximum deviation from the least-squares plane through each of the mol\u00adecules is encountered for the atoms C2A and C2B and amounts to 0.515\u2005(3) and 0.521\u2005(3)\u2005\u00c5, respectively. The almost coplanar arrangement of the aldehyde group and the pyrimidine ring in either mol\u00adecule A and B enables an intra\u00admolecular N\u2014H\u22efO inter\u00adaction (Table\u00a01S(6) graph-set motif.The asymmetric unit of the title compound contains two mol\u00adecules B Fig.\u00a02. They ar B Fig.\u00a02; an overn Table\u00a01 and form1) stack into columns parallel to [100] in an equidistant series of coplanar moieties; the independent mol\u00adecules A and B segregate into different stacks of 1.5.Mol\u00adecules of crystallizes in the non-centrosymmetric achiral space group Pna21, and its absolute structure deserves a comment. The absolute structure is linked to the direction of the polar screw axis along [001]. In the absence of heavy atoms, resonant scattering in (1) is minor, with Friedif were associated with rather large standard uncertainties: the Flack parameter and benzene rings and involve contact distances of Cg1\u22efCg3 = 3.5154\u2005(18)\u2005\u00c5 (slippage 0.954\u2005\u00c5) and of Cg7\u22efCg9 = 3.5159\u2005(19)\u2005\u00c5 (slippage 1.054\u2005\u00c5).Consecutive mol\u00adecules in each column along [100] inter\u00adact ts Fig.\u00a04. \u03c0\u2013\u03c0 stavia C=O\u22ef\u03c0 contacts; they amount to C11A=O2A\u22efCg1(x\u00a0+\u00a0y\u00a0+\u00a0z) = 3.212\u2005(2)\u2005\u00c5 and C11B=O2B\u22efCg7(x\u00a0\u2212\u00a0y\u00a0+\u00a0z) = 3.215\u2005(2)\u2005\u00c5. Perpendicular to the stacking direction, non-classical C\u2014H\u22efO hydrogen bonds , the Hirshfeld surface (HS) or longer than the sum of the van der Waals radii, respectively.In order to visualize inter\u00admolecular inter\u00adactions in (a. H\u22efH contacts are responsible for the largest contribution (49.4%) to the Hirshfeld surface . Besides these contacts, H\u22efO/O\u22efH (21.5%), H\u22efC/C\u22efH (14.9%), C\u22efC (6.7%) and N\u22efC/C\u22efN (4.0%) inter\u00adactions contribute significantly to the total Hirshfeld surface; their decomposed fingerprint plots are shown in Fig.\u00a06c\u2013f. The contributions of further contacts are only minor and amount to N\u22efO/O\u22efN (1.4%), C\u22efO/O\u22efC (1.4%), N\u22efH/H\u22efN (0.5%) and O\u22efO (0.1%).The two-dimensional fingerprint plot for all contacts is depicted in Fig.\u00a06ce Fig.\u00a06b. BesidCrystal Explorer 17 is the sum of electrostatic (Eelec), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies -one moiety with a similar planar conformation to that in the title structure: 3-(2-methyl\u00adphen\u00adyl)-2-(2-oxo\u00adphenyl\u00adeth\u00adyl)-4(3H)-quinazol\u00adinone -quinazolinone quinazolinone -quinazolinone -quinazolinone -2-[2-oxo-2-(thio\u00adphen-2-yl)ethyl\u00adidene]-3-phenyl-2,3-di\u00adhydro\u00adquin\u00adazolin-4(1H)-one , all compounds mentioned above exist as the enamine tautomer in the crystalline state, and their intra\u00admolecular N\u2014H\u22efO hydrogen bond between the ethanone and the amine N atom results in an S(6) graph set motif.A search in the Cambridge Structural Database was synthesized according to the method of Oripov et al. (1979Rf 0.78 (C6H6: MeOH 4:1). A detailed report on the synthesis of (1) and its characterization by 1H NMR is available in Zhurakulov et al. = 1.2Ueq(C). The enamine H atoms H5A and H5B were refined with a common isotropic displacement parameter; N\u2014H distances were restrained to similarity.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020016059/wm5590sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020016059/wm5590Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020016059/wm5590sup3.tifFigure S1. DOI: Click here for additional data file.10.1107/S2056989020016059/wm5590Isup4.cmlSupporting information file. DOI: 2049242CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the mol\u00adecules are linked by C\u2014H\u22efN and weak C\u2014H\u22ef\u03c0 hydrogen bonds and very weak \u03c0\u2013\u03c0 stacking inter\u00adactions. Two-dimensional fingerprint plots show that the largest contributions to the crystal stability come from H\u22efH and C\u22efH/H\u22efC inter\u00adactions. 17H16N2OS, was synthesized by a condensation reaction between 2-amino benzo\u00adthia\u00adzole and 4-N-propoxybenzaldehyde. The benzo[d]thia\u00adzole ring system is nearly planar (r.m.s. deviation 0.0088\u2005\u00c5) and makes a dihedral angle of 3.804\u2005(12)\u00b0 with the phenyl ring. The configuration about the C=N double bond is E. In the crystal structure, pairs of C\u2014H\u22efN hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions link the mol\u00adecules into inversion dimers with an R22(16) ring motif. These dimers are additionally linked by weak \u03c0\u2013\u03c0 stacking inter\u00adactions between the phenyl rings, leading to a layered arrangement parallel to (010). Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the packing arrangement are from H\u22efH (47.9%) and C\u22efH/H\u22efC (25.6%) inter\u00adactions.The title compound, C Benzo\u00adthia\u00adzole is a weak base, and is widely found in bioorganic and medicinal chemistry with application in drug discovery as a pharmacologically and biologically active compound \u2005\u00c5 for atom C4]. The benzo[d]thia\u00adzole ring system and the phenyl ring (C9\u2013C14) are slightly twisted with respect to each other, making a dihedral angle of 3.804\u2005(12)\u00b0. In the thia\u00adzole ring, the C6\u2014N1 [1.379\u2005(3)\u2005\u00c5] and C7\u2014N1 [1.288\u2005(3)\u2005\u00c5] distances indicate substantial electronic delocalization. The C8=N2 double bond has a length of 1.272\u2005(3)\u2005\u00c5, and thus is slightly longer than comparable bonds found in other Schiff base structures and the centroid of the C1\u2013C6 phenyl ring (Cg2) of an adjacent mol\u00adecule : \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a02, \u2212z) where Cg3 is the centroid of the C9\u2013C14 phenyl ring. The resulting supra\u00admolecular network is layered and expands parallel to (010).In the crystal structure, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 hydrogen bonds Table\u00a01 between le Fig.\u00a02. Pairs oif Fig.\u00a02. The dimCrystalExplorer -N-benzyl\u00adidenebenzo[d]thia\u00adzol-2-amine skeleton gave 20 hits. Of these 20, the most similar to the title compound are 2-[carbonoimido\u00adyl]phenol -2-[(6-eth\u00adoxy\u00adbenzo\u00adthia\u00adzol-2-yl)imino\u00admeth\u00adyl]-6-meth\u00adoxy\u00adphenol was then added dropwise into the mixture under stirring, in the presence of a catalytic amount of glacial acetic acid. The reaction mixture was then placed inside an unmodified household microwave oven and was irradiated for 32\u2005min (eight pulses each of 4\u2005min) at 540\u2005W power, with short inter\u00adruptions of one minute. The progress of the reaction was monitored by thin-layer chromatography using ethyl acetate and n-hexane (3:7 v:v) as eluent (Rf = 0.69). The formed precipitate was filtered off, washed with 1-propanol, and dried. The resulting solid was further purified by recrystallization from n-hexane to give the pure imine as a crystalline solid .2-Amino benzo\u00adthia\u00adzole was dissolved in 10\u2005ml of 1-propanol in a 50\u2005ml borosilicate glass beaker. 4-Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other C\u2013bound H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020012128/wm5582sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020012128/wm5582Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020012128/wm5582Isup3.cmlSupporting information file. DOI: 1979807CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Diels\u2013Alder cyclo\u00adaddition of cyclo\u00adhepta\u00adtriene and maleic anhydride produces the title carb\u00adoxy\u00adlic anhydride; reaction of this anhydride with 4-bromo\u00adphenyl\u00adaniline forms the corresponding tetra\u00adcyclic imide. The anhydride features C\u2014H\u22efO hydrogen bonds in the solid state, while the imide also features C\u2014H\u22efO hydrogen bonds as well as C\u2014H\u22ef\u03c0 and lone pair\u2013\u03c0 inter\u00adactions. 11H10O3 (I) and C17H14BrNO2 (II), both containing the bi\u00adcyclo\u00ad[2.2.2]octene ring system, are reported here octene ring system. Non-covalent inter\u00adactions present in the crystal structure of II include a number of C\u2014H\u22efO inter\u00adactions. The extended structure of II also features C\u2014H\u22efO hydrogen bonds as well as C\u2014H\u22ef\u03c0 and lone pair\u2013\u03c0 inter\u00adactions, which combine together to create supra\u00admolecular sheets.The syntheses and crystal structures of the two title compounds, C Goh 2014. Private The bond lengths of the carbonyl groups of the anhydride are shorter than the imide, as expected, with C1=O1 = 1.1943\u2005(18), C2=O2 = 1.1904\u2005(17), C1\u2014O3 = 1.3868\u2005(17) and C2\u2014O3 = 1.3978\u2005(16)\u2005\u00c5. The corresponding data for the C1a mol\u00adecule are 1.1913\u2005(17), 1.1871\u2005(18), 1.3855\u2005(17) and 1.3905\u2005(18)\u2005\u00c5, respectively. The configurations of the stereogenic centres in the arbitrarily chosen asymmetric mol\u00adecules are: C3 S, C4 R, C5 R, C8 S, C9 S, C10 R and C3a R, C4a S, C5a S, C8a R, C9a R, C10a S: crystal symmetry generates a racemic mixture in the bulk.The structure of the title anhydride II was solved in the monoclinic space group P21/n, and its atom labeling scheme is shown in Fig.\u00a04II are C3 R, C4 S, C5 S, C8 R, C9 R and C10 S; again, crystal symmetry generates a racemic mixture.The structure of the imide I is dominated by C\u2014H\u22efO hydrogen bonds to 3.4882\u2005(17)\u2005\u00c5 with D\u2014H\u22efA angles ranging from 119 to 159\u00b0; the C9 bond is likely very weak based on its H\u22efA distance of 2.73\u2005\u00c5. Combined together, these inter\u00adactions create supra\u00admolecular sheets that lie in the ab plane.The extended structure of the anhydride II, the mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds as well as C\u2014H\u22ef\u03c0 and C\u2014Br\u22ef\u03c0 inter\u00adactions (Table\u00a02D\u22efA distance of 3.175\u2005(2)\u2005\u00c5 with a D\u2014H\u22efA angle of 139\u00b0. The C\u2014H\u22ef\u03c0 inter\u00adaction is between C3\u2014H3, which is \u03b1 to the carbonyl group C1(O1), and the aromatic ring C12\u2013C17. This inter\u00adaction has a H\u22efCg distance of 3.801\u2005(2)\u2005\u00c5 (where Cg is the centroid of the C12\u2013C17 ring), with a C\u2014H\u22efCg angle of 165\u00b0. The aromatic ring C12\u2013C17 bears an electron-withdrawing bromine atom, and accepts a lone pair(LP)\u2013\u03c0 inter\u00adaction from the bromine atom of a nearby mol\u00adecule \u2005\u00c5 with a C15\u2014Br1\u22efCg angle of 87.43\u2005(6)\u00b0. Dimers of imide II are formed via the Br\u22ef\u03c0 inter\u00adactions, and these dimers are linked into supra\u00admolecular sheets that lie along (010) by the C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions via short-path distillation and the reaction mixture was left to cool at room temperature. The round-bottom flask was fitted with a stopper and left to recrystallize for 48\u2005h to afford large, cream-colored needles. The product was recrystallized once more by dissolving in 8\u2005ml of xylene: after a week at room temperature, the pure product I was obtained in the form of large colorless crystals . 1H NMR \u03b4 5.88 , 3.46 , 3.23 , 1.17\u20131.04 . 13C NMR \u03b4 172.45, 128.55, 45.88, 33.65, 9.56, 5.24.Cyclo\u00adhepta\u00adtriene and maleic anhydride were added to an oven-dried round-bottom flask containing 10\u2005ml of xylene and the mixture was refluxed for 1.5\u2005h. Approximately half of the xylenes were distilled off Synthesis of the imide (II)I and p-bromo\u00adaniline were added to a vial containing 5\u2005ml of xylene and the mixture was refluxed for 5\u2005min. The mixture was then cooled to room temperature and left for 5 days in a sealed vial. The precipitate was recrystallized from ethanol solution to yield colorless needle-like crystals of II . 1H NMR \u03b4 7.54 , 7.06 , 5.84 , 3.48 , 3.12 , 1.14 , 0.38\u20130.21 . 13C NMR \u03b4 177.42, 132.34, 130.88, 128.11, 127.92, 122.47, 45.40, 33.90, 9.97, 4.80.Compound Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989020009512/hb7931sup1.cifCrystal structure: contains datablock(s) global, II, I. DOI: 10.1107/S2056989020009512/hb7931Isup3.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989020009512/hb7931IIsup4.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989020009512/hb7931Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020009512/hb7931IIsup5.cmlSupporting information file. DOI: 2015807, 2015806CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "It should read as \u201cbuffer A (10 mM Tris\u2013HCl (pH 8.0), 50 mM NaCl, 1 mM NaThe authors would like to apologize for any inconvenience caused."} +{"text": "The title di-substituted thio\u00adurea has hy\u00addroxy\u00adlethyl and ethyl benzoate substituents bound to the same amine-N atom; overall the mol\u00adecule is twisted. Supra\u00admolecular layers are formed in the crystal, with the mol\u00adecules connected by O\u2014H\u22efS and N\u2014H\u22efO hydrogen bonds. 12H16N2O3S, has the hy\u00addroxy\u00adlethyl and ethyl benzoate substituents bound to the same amine-N atom, and is twisted, having a (+)syn-clinal conformation with the Namine\u2014C\u2014C\u2014O torsion angles of 49.39\u2005(13) and 59.09\u2005(12)\u00b0, respectively; the dihedral angle between the almost planar CN2S core and the pendent benzene ring is 69.26\u2005(4)\u00b0. In the crystal, supra\u00admolecular layers propagating in the ac plane are formed via a combination of hydroxyl-O\u2014H\u22efS(thione), amine-N\u2014H\u22efO hydrogen-bonds. The layers stack along the b axis with inter-digitation of the benzene rings allowing the formation of \u03c0\u2013\u03c0 stacking [inter-centroid separation = 3.8722\u2005(7)\u2005\u00c5] and parallel C=O\u22ef\u03c0 inter\u00adactions. A computational chemistry study shows the conventional hydrogen bonding in the crystal leads to significant electrostatic stabilization but dispersion terms are also apparent, notably through the inter\u00adactions involving the benzene residue.The title di-substituted thio\u00adurea, C In an experiment with R = CH2CH2OH and Ar = C6H5, the solution was also heated resulting in an apparent rearrangement with deprotonation of one hy\u00addroxy\u00adethyl group followed by nucleophilic attachment at the carbonyl-C atom along with protonation of the primary amine and cleavage of the original N\u2014C(=O) bond to yield (I)2NC(=S)N(CH2CH2OH)CH2CH2OC(=O)C6H5. The mol\u00adecular structure of (I)The title compound, (I)2S atoms of the thio\u00adurea core are almost planar, exhibiting a r.m.s. deviation = 0.0054\u2005\u00c5, with the appended C2 and C4 atoms lying 0.0236\u2005(18) and 0.0216\u2005(16)\u2005\u00c5 to either side of the plane. The conformation of the C2-hy\u00addroxy\u00adlethyl residue is (+)syn-clinal as indicated by the N2\u2014C2\u2014C3\u2014O1 torsion angle of 49.39\u2005(13)\u00b0. The CO2 residue is close to co-planar with the (C7\u2013C12)-benzene ring to which it is connected, forming a dihedral angle of 4.83\u2005(9)\u00b0. The dihedral angle between the least-squares planes through the CN2S core and the benzene ring is 69.26\u2005(4)\u00b0, indicating the mol\u00adecule is highly twisted. Finally, the N2\u2014C4\u2014C5\u2014O2 torsion angle of 59.09\u2005(12)\u00b0 is indicative of a (+)syn-clinal configuration about the C\u2014C bond, thereby confirming the twisted nature of the mol\u00adecule.The mol\u00adecule of (I)a-axis direction, Fig.\u00a02a). These hydrogen bonds also lead to the formation of 12-membered {\u22efHO\u22efHNCS}2 and 14-membered {\u22efOC2NCNH}2 synthons, each disposed about a centre of inversion, and linked via the edges defined by the amine-N\u2014H\u22efO(hydrox\u00adyl) hydrogen bonds. The tape has a step-ladder topology and projecting laterally to either side of the tape are the remaining amine-H and carbonyl-O atoms, which form the donors and acceptors of amine-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds to link the tapes into a layer in the ac plane, Fig.\u00a02b). The directional links between layers are twofold, namely \u03c0\u2013\u03c0 stacking between the centrosymmetrically related benzene rings and parallel C=O\u22ef\u03c0 inter\u00adactions, Table\u00a01c). These inter\u00adactions are possible owing to the inter-digitation of the benzene rings along the b-axis direction, as highlighted in Fig.\u00a02d).As anti\u00adcipated, hydrogen bonding plays a key role in the supra\u00admolecular assembly of (I)Crystal Explorer 17 hydrogen bond with a dnorm distance of 1.92\u2005\u00c5, which is significantly shorter, by 0.69\u2005\u00c5 , the ester-C6\u22ef\u03c0(benzene), benzene-C9\u2014H9\u22efS1(thione) and benzene-C9\u2013H9\u22efC1(thione) contacts with a combined Eint of \u221248.3\u2005kJ\u2005mol\u22121 and hydroxyl-O1\u2013H1O\u22efS1(thione) [\u221228.8\u2005kJ\u2005mol\u22121]. Close in energy to latter is that due to \u03c0\u2013\u03c0 [Cg1\u22efCg1 = 3.8722\u2005(7)\u2005\u00c5] with Eint = \u221228.3\u2005kJ\u2005mol\u22121. Next most significant are the pairwise ethyl\u00adene-C2\u2014H2A\u22efS1(thione) inter\u00adactions (Eint = \u221223.1\u2005kJ\u2005mol\u22121) then methyl\u00adene-C3\u2014H3B\u22efH8(benzene) (Eint = \u22124.3\u2005kJ\u2005mol\u22121).Among all the inter\u00adactions, it is the amine-N1\u2014H2viz. amine-N1\u2014H1N\u22efO3(carbon\u00adyl) that propagates along the c axis together with amine-N1\u2014H2N\u22efO1(hydrox\u00adyl) and hydroxyl-O1\u2014H1O\u22efS1(thione), which extend along the a axis, thereby forming a step-ladder framework as shown in Fig.\u00a07a). On the other hand, significant dispersion force is also present as evidenced from the wire mesh-like dispersion energy framework predominantly governed by the \u03c0\u2013\u03c0 inter\u00adactions, with contributions from the inter\u00adactions involving the benzene-C9 atom, Fig.\u00a07b). Overall, the combination of electrostatic and dispersion forces leads to a cuboid-like framework shown in Fig.\u00a07c).The crystal of (I)R(R\u2032)NC(=S)NH2 are comparatively rare with the simplest derivative being the R = R\u2032 = Me species, the almost planar mol\u00adecule being first reported in 1994 : 3419 \u03bd(OH), 3323 \u03bd(NH2)asym, 3222 \u03bd(NH2)sym, 3058 \u03bd(CH)arom, 3002\u20132881 \u03bd(CH), 1706 \u03bd(COO), 1647 \u03bd(C=O), 1600 \u03b4(NH), 1523 \u03bd(C=C), 1270 \u03bd(CN), 1053 \u03bd(C=S), 711 \u03b4(CH).Compound (I)Uiso(H) set to 1.2Ueq(C). The oxygen- and nitro\u00adgen-bound H atoms were located from a difference-Fourier map and refined with O\u2014H = 0.84\u00b10.01\u2005\u00c5 and N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.5Ueq(O) or 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989020006829/hb7918sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020006829/hb7918Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020006829/hb7918Isup3.cmlSupporting information file. DOI: 2004940CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A study of their luminescence properties indicates that co-crystals 1 and 2 exhibit distinctly different luminescence in the solid state at room temperature.Two new co-crystals of tetra\u00adiodo\u00adethyl\u00adene with two aza\u00adphenanthrenes were successfully synthesized. In the crystals, C\u2014I\u22ef 2I4\u00b7C13H9N (1) and tetra\u00adiodo\u00adethyl\u00adene\u2013benzo[f]quinoline (1/2), 0.5C2I4\u00b7C13H9N (2), were obtained from tetra\u00adiodo\u00adethyl\u00adene and aza\u00adphenanthrenes, and characterized by IR and fluorescence spectroscopy, elemental analysis and X-ray crystallography. In the crystal structures, C\u2014I\u22ef\u03c0 and C\u2014I\u22efN halogen bonds link the independent mol\u00adecules into one-dimensional chains and two-dimensional networks with subloops. In addition, the planar aza\u00adphenanthrenes lend themselves to \u03c0\u2013\u03c0 stacking and C\u2014H\u22ef\u03c0 inter\u00adactions, leading to a diversity of supra\u00admolecular three-dimensional structural motifs being formed by these inter\u00adactions. Luminescence studies show that co-crystals 1 and 2 exhibit distinctly different luminescence properties in the solid state at room temperature.Two new co-crystals, tetra\u00adiodo\u00adethyl\u00adene\u2013phenanthridine (1/2), 0.5C Halogen bonding (XB) is a powerful tool to assemble supra\u00admolecular materials and to promote chemical or biological mol\u00adecular recognition , which is expected to tune their luminescence behaviour via a change of the co-crystal structures. Single crystal X-ray diffraction (XRD) data reveal that the two co-crystals of TIE with PHN and BfQ reported here have inter\u00adesting structural properties and exhibit different luminescence behaviour from previous reports. TIE as a quadridentate XB donor allows the formation of three-dimensional halogen-bonded networks with XB acceptors, PHN and BfQ. Using the conventional solution-based method, yellow co-crystals suitable for XRD measurement were obtained. The crystal structures of the co-crystals are mainly constructed by C\u2014I\u22ef\u03c0 and C\u2014I\u22efN halogen bonds. Other multiple inter\u00admolecular inter\u00adactions, such as \u03c0\u2013\u03c0 stacking, C\u2014H\u22ef\u03c0, C\u2014H\u22efI as well as C\u2014H\u22efH\u2014C inter\u00adactions, are also observed in the co-crystals.Nitro\u00adgen heteroaromatic rings are a common type of luminescence or luminescent precursor materials. However, in general, it is difficult to use them to generate phospho\u00adrescence or delayed fluorescence. Haloperfluoro\u00adbenzenes, as XB donors, have been used in attempts to assemble luminescence co-crystals with aza\u00adphenanthrenes . The I1\u22efN1i distance is 2.864\u2005(7)\u2005\u00c5 and the corresponding C14\u2013I1\u22efN1i angle is 172.8\u2005(2)\u00b0 . The strong C14\u2014I1\u22efN1 halogen bond results in a I1\u22efC13i distance [3.553\u2005(8)\u2005\u00c5] shorter than the sum of the van der Waals radii, which indicates a C1\u2014I1\u22efC13 halogen inter\u00adaction. In addition, the C14\u2014I2\u22efC9ii/C10ii C\u2014I\u22ef\u03c0 separations are 3.432\u2005(9) and 3.612\u2005(8)\u2005\u00c5, and the corresponding bond angles are 165.6\u2005(2) and 156.7\u2005(2)\u00b0, respectively. Furthermore, \u03c0\u2013\u03c0 stacking and C\u2014H\u22efH\u2014C inter\u00adactions between two adjacent PHN mol\u00adecules contribute to the extension of the two-dimensional network into a three-dimensional supra\u00admolecular structure , I1\u22efC1 = 3.641\u2005(5), I2\u22efC13 = 3.436\u2005(5) and I2\u22efC8 = 3.733\u2005(4)\u2005\u00c5, respectively] in co-crystal 2 are all a little longer (0.004\u20130.121\u2005\u00c5) than in 1 . In addition, the two-dimensional network \u2005\u00c5, Cg1\u22efCg2ii = 3.963\u2005(2)\u2005\u00c5, Cg1\u22efCg3ii = 3.746\u2005(3)\u2005\u00c5, Cg2\u22efCg2ii = 3.768\u2005(2)\u2005\u00c5; Cg1, Cg2 and Cg3 are the centroids of rings N1/C1\u2013C5, C4\u2013C9 and C8\u2013C13, respectively; symmetry codes: (i) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z; (ii) 1\u00a0\u2212\u00a0x, \u2212y, 1\u00a0\u2212\u00a0z] and C\u2014H\u22efI hydrogen bonds in the 2\u03b8 range of 5 to 50\u00b0. As shown in Fig.\u00a061 and 2 match well with the spectra simulated from the XRD data, which confirms the purity of 1 and 2.The powder X-ray diffraction (PXRD) experiments were carried out for the title co-crystals using a Bruker D8-ADVANCE X-ray diffractometer state to drop below that of the 3 state. It is supposed that for the singlet states the 0\u20130 transition of emitters in co-crystals is localized at 375\u2005nm and 450\u2005nm, respectively, and for triplet states the 0\u20130 transition is at about 600\u2005nm. The energy gap between S1 and T1 is largely greater than 20\u2005kJ\u2005mol\u22121, so the delayed fluorescence most likely originates from the triplet\u2013triplet annihil\u00adation process, named P-type delayed fluorescence (P-DF). Both delayed fluorescence and phospho\u00adrescence are relative to triplet states, so they should be significant for improving the exciton emission efficiency of luminescence materials are about 10\u2005ns, while the delayed fluorescence lifetime (4.36\u2005\u00b5s for 1 and 6.45\u2005\u00b5s for 2) is less than the 10\u2005\u00b5s level because of the strong heavy-atom effect leading to a faster decay of the triplet state. Additionally, the phospho\u00adrescence is too weak to measure its decay lifetime. However, the phospho\u00adrescence lifetime can be estimated to be about 20\u2005\u00b5s based on the relationship between P-DF and the accompanying phospho\u00adrescence measurements were obtained by slow evaporation of the solvent at room temperature after about two weeks. Elemental analysis calculated for C14H9NI2 (445.02): C 37.78, H 2.04, N 3.15. Found: C 37.54, H 2.31, N 3.26. For co-crystal 1, and C 37.85, H 2.16, N 3.04 for co-crystal 2. IR For 1: 3048(w), 1603(w), 1572(w), 1494(m), 1446(w), 1382(m), 1293(m), 1267(m), 1189(m), 1089(m), 948(m), 870(s), 832(s), 802(s), 749(s), 707(s), 615(m), 538(m), 487(m), 435(m). For 2: 3048(w), 1611(w), 1576(s), 1522(w), 1486(w), 1458(m), 1440(m), 1238(m), 1132(m), 1032(m), 953(m), 924(m), 889(s), 745(s), 714(s), 610(m), 552(m), 448(m), 423(m).0.1\u2005mmol of PHN/BfQ and 0.05\u2005mmol of TIE were dissolved in an acetone/chloro\u00adform (2:1) mixture in a glass vial. Well\u2013formed co-crystals Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989020002182/vm2225sup1.cifCrystal structure: contains datablock(s) 1, 2. DOI: 10.1107/S2056989020002182/vm22251sup4.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989020002182/vm22252sup5.hklStructure factors: contains datablock(s) 2. DOI: 1899078, 1899077CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The three para-phenyl\u00adene rings bonded to the N atom are in a propeller form. All of the naphthalene ring systems are slightly bent. In the crystal, mol\u00adecules form an inversion dimer, through two pairs of C\u2014H\u22ef\u03c0 inter\u00adactions, which further inter\u00adacts with the adjacent dimer via another two pairs of C\u2014H\u22ef\u03c0 inter\u00adactions, forming a column structure along the a axis. There are no significant inter\u00adactions between these column structures.In the title mol\u00adecule, C The torsion angles C3\u2014C2\u2014N1\u2014C34, C19\u2014C18\u2014N1\u2014C2 and C35\u2014C34\u2014N1\u2014C18 are \u221235.0\u2005(2), \u221260.6\u2005(2) and \u221230.3\u2005(2)\u00b0, respectively. The para-phenyl\u00adene ring and the mean plane of the neighboring naphthalene ring system are inclined to each other by 54.66\u2005(7)\u00b0 for (C2\u2013C7)/(C8\u2013C17), 48.80\u2005(7)\u00b0 for (C18\u2013C23)/(C24\u2013C33) and 56.21\u2005(7)\u00b0 for (C34\u2013C39)/(C40\u2013C49).The mol\u00adecular structure of the title compound is shown in Fig.\u00a01via four inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions \u2212x\u00a0+\u00a01, \u2212y, \u2212z\u00a0+\u00a02], forming an inversion dimer \u2212x, \u2212y, \u2212z\u00a0+\u00a02] , which included eleven hits (nine compounds) with heteroaromatic rings and eight hits (seven compounds) with phenyl rings. The seven compounds with phenyl rings at the three para-position of the tri\u00adphenyl\u00adamine skeleton include tris\u00ad(biphenyl-4-yl)amine [WEHLIE phen\u00adyl]amine , toluene (42\u2005mL) and water (10.4\u2005mL) were placed in a 100\u2005mL round-bottom flask. After the solution was purged with nitro\u00adgen for 10 minutes, it was heated at 373\u2005K under nitro\u00adgen for 24\u2005h. The reaction mixture was extracted with ethyl acetate. After drying over anhydrous Na2SO4, the organic layer was evaporated. The residue was redissolved in a small amount of ethyl acetate. The addition of a large amount of methanol gave the pure product as a white precipitate . Colorless single crystals suitable for X-ray diffraction were obtained by means of the vapor diffusion method from chloro\u00adform as a rich solvent and n-hexane as a poor solvent after standing for one week.The title compound was prepared by a modification of the previously reported Suzuki\u2013Miyaura coupling reaction (Kwon Uiso(H) = 1.2Ueq(C). One outlier (011) was omitted from the refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020012529/is5556sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989020012529/is5556Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020012529/is5556Isup3.cmlSupporting information file. DOI: 2031765CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Biological screening showed these compounds as good inhibitors for several glycosidases. Especially Glycosidases are involved in many important biological processes, and inhibitors of glycosidases \u20133 are diAn extended search in literature as of 2020, October, revealed there are some 30,600 structures containing the substructure of a 6-amino-6-deoxy-hexopyranose (or -pyranoside). Monofluorinated analogs, however, have scarcely been prepared \u201337\u2014but t1 D\u2009=\u2009+39.9\u00b0 ]; R\u0192 \u2009=\u20090.53; analysis calcd for C23H26O6 (398.45): C 69.33, H 6.58; found: C 69.09, H 6.63.Data for C ; R\u0192 \u2009=\u20090.63; analysis calcd for C23H26O6 (398.45): C 69.33, H 6.58; found: C 69.17, H 6.71.Data for C (lit.: 124\u2009\u00b0C);1 in dry DCM (40.0\u2009mL) while maintaining the temperature at \u221278\u2009\u00b0C. The mixture was stirred for 2\u2009h at \u221278\u2009\u00b0C, then a solution of Et3N in dry DCM (40.0\u2009mL) was added dropwise. The mixture was stirred at \u221278\u2009\u00b0C for another 30\u2009min and at 25\u2009\u00b0C for 12\u2009h. Usual aqueous workup followed by chromatography gave 2 as a white solid; MP 130\u2013132\u2009\u00b0C; [\u03b1]D\u2009=\u2009\u221247.95\u00b0 ; R\u0192 \u2009=\u20090.40 (ketone); R\u0192 \u2009=\u20090.19 (hydrate); analysis calcd for C23H24O6 (396.43): C 69.68, H 6.10; found: C 69.43, H 6.27.To a solution of dry DMSO in dry DCM (40.0\u2009mL) at \u221278\u2009\u00b0C, a solution of trifluoroacetic anhydride in dry DCM (10.0\u2009mL) was slowly added dropwise, and the mixture was stirred at this temperature for 45\u2009min followed by adding a solution of 2 in dry DCM (6.0\u2009mL), DAST (417\u2009\u03bcL 3.03\u2009mmol) was added dropwise under argon, and the mixture was stirred at 25\u2009\u00b0C for 5 days. Methanol (1.0\u2009mL) was carefully added, and the solvents were removed under diminished pressure. The oily residue was dissolved in DCM (90\u2009mL) and washed with water (50\u2009mL). The aqueous phase was re-extracted with DCM (3\u2009\u00d7\u2009100\u2009L); the organic layers were combined, and the solvent was evaporated under reduced pressure. The remaining residue was subjected to chromatography to afford 3 as a white solid; MP 79\u201381\u2009\u00b0C; [\u03b1]D\u2009=\u2009\u221226.10\u00b0 R\u0192 \u2009=\u20090.75; analysis calcd for C23H24O5F2 (418.43): C 66.02, H 5.78; found: C 65.85, H 5.92.To a solution of 3 in dry ether (30\u2009mL) and THF (30\u2009mL), lithium aluminum hydride was added in several portions, and the suspension was stirred for 15\u2009min at 25\u2009\u00b0C. Then, the suspension was heated to reflux, and a solution of dry aluminum chloride in dry ether (30\u2009mL) was added dropwise followed by stirring under reflux for another 48\u2009h. The suspension was cooled to 25\u2009\u00b0C, methanol (30\u2009mL) was carefully added, and stirring was continued for another 30\u2009min. After usual aqueous workup followed by chromatography 4 and 5 were obtained each as a colorless oil.To an ice-cold solution of 4: [\u03b1]D\u2009=\u2009\u221248.23\u00b0 ; R\u0192 \u2009=\u20090.51; analysis calcd for C23H26O5F2 (420.45): C 65.70, H 6.23; found: C 65.55, H 6.40.Data for 5: [\u03b1]D\u2009=\u2009\u221218.27\u00b0 ; R\u0192 \u2009=\u20090.41; analysis calcd for C23H26O5F2 (420.45): C 65.70, H 6.23; found: C 65.47, H 6.39.Data for 5 in toluene (30\u2009mL) containing triphenylphosphane and imidazole , iodine was added in several portions. After stirring at 90\u2009\u00b0C for 2\u2009h, the reaction mixture was decanted, and the remaining oil was washed with ether (3\u2009\u00d7\u2009100\u2009mL). The combined organic layers were evaporated, and the remaining residue was subjected to chromatography to afford 6 as a colorless oil; [\u03b1]D\u2009=\u2009\u221225.32\u00b0 ; R\u0192 \u2009=\u20090.56; analysis calcd for C23H25O4F2I (530.34): C 52.09, H 4.75; found: C 51.84, H 4.92.To a solution of 6 in dry DMF (19\u2009mL) a solution of lithium azide was added at 25\u2009\u00b0C, and the solution was stirred at this temperature for 4 days. The solvents were removed under diminished pressure, and the remaining residue was dissolved in DCM (100\u2009mL) and water (50\u2009mL). The aq. phase was extracted with DCM (3\u2009\u00d7\u2009100\u2009mL), and the combined organic layers were dried (Na2SO4). The solvent was removed under reduced pressure, and the remaining residue was subjected to chromatography to afford 7 as a colorless oil; [\u03b1]D\u2009=\u2009\u221233.01\u00b0 ; R\u0192 \u2009=\u20090.39; analysis calcd for C23H25O4F2N3 (445.44): C 62.01, H 5.66, N 9.43; found: C 61.80, H 5.81, N 9.25.To a solution of 7 in dry MeOH (20.0\u2009mL) containing palladium on charcoal was hydrogenated . The solution was filtered through a pad of Celite; the pad was rinsed with methanol (4\u2009\u00d7\u200950\u2009mL). The combined organic layers were dried (MgSO4), the solvent was removed under reduced pressure, and the residue was subjected to chromatography to afford 8 as a white foam; [\u03b1]D\u2009=\u2009\u221220.26\u00b0 ; R\u0192 \u2009=\u20090.14; analysis calcd for C9H17O4F2N (241.23): C 44.81, H 7.10, N 5.81; found: C 44.69, H 7.32, N 5.69.A solution of 9 in dry dichloromethane (40.0\u2009mL) was added dropwise, maintaining the temperature at \u221278\u2009\u00b0C during this addition. The mixture was stirred for 2\u2009h at \u221278\u2009\u00b0C, then a solution of Et3N in dry DCM (40.0\u2009ml) was added. The mixture was stirred at \u221278\u2009\u00b0C for another 30\u2009min and at 25\u2009\u00b0C for 12\u2009h. Usual aqueous workup followed by chromatography gave 10 as a white solid; MP 125\u2013127\u2009\u00b0C; [\u03b1]D\u2009=\u2009\u221264.84\u00b0 ; R\u0192 \u2009=\u20090.73; analysis calcd for C23H24O6 (396.43): C 69.68, H 6.10; found: C 69.50, H 6.37.To a mixture of dry DMSO and dry DCM (72.0\u2009ml) at \u221278\u2009\u00b0C, a solution of trifluoroacetic anhydride in dry DCM (18.0\u2009mL) was slowly added, and the mixture was stirred at this temperature for 45\u2009min. Then a solution of 10 in dry DCM (6.0\u2009mL) DAST was added dropwise, and the mixture was stirred at 25\u2009\u00b0C for 5 days. Methanol (2.0\u2009mL) was carefully added, and the solvents were removed under diminished pressure. Usual aqueous workup followed by chromatography gave 11 as a white amorphous solid; [\u03b1]D\u2009=\u2009\u221227.89\u00b0 ; R\u0192 \u2009=\u20090.67; analysis calcd for C23H24O5F2 (418.43): C 66.02, H 5.78; H 65.79, H 5.93.To a solution of 11 in dry ether (40\u2009mL) and THF (40\u2009mL) lithium aluminum hydride was added in several portions, and the suspension was stirred for 15\u2009min at 25\u2009\u00b0C. Then, the suspension was heated under reflux, and a solution of dry aluminum chloride in dry ether (30\u2009mL) was added followed by stirring under reflux for 72\u2009h. The suspension was cooled to 25\u2009\u00b0C, methanol (30\u2009mL) was carefully added, and stirring was continued for another 45\u2009min. Usual aqueous workup followed by chromatography gave 12 and 13 each as a viscous oil;To an ice-cold solution of 12: [\u03b1]D\u2009=\u2009+14.57\u00b0 ; R\u0192 \u2009=\u20090.47; analysis calcd for C23H26O5F2 (420.45): C 65.70, H 6.23; found: C 65.59, H 6.46.Data for 13: [\u03b1]D\u2009=\u2009\u221217.26\u00b0 ; R\u0192 \u2009=\u20090.61; analysis calcd for C23H26O5F2 (420.45): C 65.70, H 6.23; found: C 65.56, H 6.41.Data for 12 in toluene (40\u2009mL) containing triphenylphosphane and imidazole iodine was added in several portions. After stirring at 90\u2009\u00b0C for 1\u2009h, the reaction mixture was decanted and the remaining oil was washed with ether (4\u2009\u00d7\u2009100\u2009mL). The combined organic layers were evaporated, and the remaining residue was subjected to chromatography to afford 14 as a colorless oil; [\u03b1]D\u2009=\u2009+23.79\u00b0 ; R\u0192 \u2009=\u20090.51; analysis calcd for C23H25O4F2I (530.34): C 52.09, H 4.75; found: C 51.84, H 4.93.To a solution of 14 in dry DMF (29\u2009mL) a solution of lithium azide was added at 25\u2009\u00b0C, and the solution was stirred at this temperature for 4 days. The solvents were removed under diminished pressure, and the remaining residue was dissolved in DCM (100\u2009mL) and water (50\u2009mL). The aq. phase was extracted with DCM (3\u2009\u00d7\u2009100\u2009ml), and the combined organic layers were dried (Na2SO4). The solvent was removed under reduced pressure, and the remaining residue was subjected to chromatography to afford 15 as a colorless oil; [\u03b1]D\u2009=\u2009+33.95\u00b0 ; R\u0192 \u2009=\u20090.48; analysis calcd for C23H25O4F2N3 (445.44): C 62.01, H 5.66, N 9.43; found: C 61.86, H 5.71, N 9.21.To a solution of 15 in dry methanol (20.0\u2009mL) containing palladium on charcoal was hydrogenated . Workup as described above followed by chromatography gave 16 as a white foam; [\u03b1]D\u2009=\u2009\u221227.95\u00b0 ; R\u0192 \u2009=\u20090.13; analysis calcd for C9H17O4F2N (241.23): C 44.81, H 7.10, N 5.81; found: C 44.65, H 7.34, N 5.68.A solution of Supplementary Materials"} +{"text": "Scientific Reports 10.1038/srep45179, published online 23 March 2017Correction to: 3 NFs synthesized at 210\u2009\u00b0C for 12\u2009h shown in panels d-f were incorrectly given as SEM images of P(VDF-HFP) based nanocomposites with 5.0\u2009vol%, 10.0\u2009vol% and 20.0\u2009vol% BaTiO3 NFs synthesized at 210\u2009\u00b0C for 24\u2009h. The correct Figure 5 appears below as Fig.\u00a0This article contains errors in Figure 5. The SEM images of the nanocomposites with 2.5\u2009vol%, 5.0\u2009vol% and 7.5\u2009vol% BaTiO"} +{"text": "Corrections to the article, \u201cMultilabel segmentation of cancer cell culture on vascular structures with deep neural networks,\u201d by S. Rahkonen et\u00a0al. J. Med. Imag.7(2), 024001 (2020) doi: 10.1117/1.JMI.7.2.024001] was originally published online on 7 April 2020 with errors in the panel labels for Fig.\u00a06.This article [The following corrections were made to the originally published version:1.in Fig 6(h), \u201cPrediction (O)\u201d was corrected to read \u201cPrediction (S)\u201d;2.in Fig 6(i), \u201cPrediction (I)\u201d was corrected to read \u201cPrediction (IP)\u201d;3.in Fig 6(j), \u201cErrors (R)\u201d was corrected to read \u201cErrors (P)\u201d;4.in Fig 6(k), \u201cErrors (O)\u201d was corrected to read \u201cErrors (S)\u201d;5.in Fig 6(l), \u201cErrors (I)\u201d was corrected to read \u201cErrors (IP)\u201d;6.in Fig 6(m), \u201cAbs. errors (R)\u201d was corrected to read \u201cAbs. errors (P)\u201d;7.in Fig 6(n), \u201cAbs. errors (O)\u201d was corrected to read \u201cAbs. errors (S)\u201d; and8.in Fig 6(o), \u201cAbs. errors (I)\u201d was corrected to read \u201cAbs. errors (IP).\u201dThis article was corrected on 13 April 2020."} +{"text": "MF6 compounds and a related coordination polymer are reported.The crystal structures of three copper(II)-bi\u00adpyridine\u2013 2N,N\u2032)copper(II) hexa\u00adfluorido\u00adsilicate tetra\u00adhydrate, [Cu(bpy)2(H2O)][SiF6]\u00b74H2O , (I), bis\u00ad-di-\u03bc-fluorido-1:3\u03ba2F:F;2:3\u03ba2F:F-deca\u00adfluorido-1\u03ba5F,2\u03ba5F-ditantalum(V)copper(II), [Cu(bpy)2(TaF6)2], (II), tris\u00adcopper(II) bis[hexa\u00adfluorido\u00adtantalate(V)], [Cu(bpy)3][TaF6]2, (III), and catena-poly[[di\u00adaqua\u00adcopper(II)]-\u03bc-fluorido-tetra\u00adfluorido\u00adtin-\u03bc-fluorido], [Cu(bpy)(H2O)2SnF6]n, (IV). Compounds (I), (II) and (III) contain locally chiral copper coordination complexes with C2, D2, and D3 symmetry, respectively. The extended structures of (I) and (IV) are consolidated by O\u2014H\u22efF and O\u2014H\u22efO hydrogen bonds. The structure of (III) was found to be a merohedral (racemic) twin.We report the hydro\u00adthermal syntheses and crystal structures of aqua\u00adbis\u00ad(2,2\u2032-bi\u00adpyridine-\u03ba The structure features isolated C2-symmetric \u0394- and \u039b-[Cu(bpy)2(H2O)]2+ cations and octa\u00adhedral SiF62\u2013 anions , as described by the parameter \u03c4 = (\u03b2\u00a0\u2212\u00a0\u03b1)/60, where \u03b2 and \u03b1 are the two largest angles of the complex 2(H2O)]2+ complexes ns Fig.\u00a01. The fiv2(TaF6)2 and crystallizes in space group P2(TaF6)2 complexes with local D2 symmetry. Each CuII center is equatorially coordinated by two bpy ligands and axially coordinated by two TaF6\u2212 groups. Two independent Cu(bpy)2(TaF6)2 units with the same handedness are present within the arbitrarily chosen asymmetric unit it Fig.\u00a02. These c3][TaF6]2 and crystallizes in the enanti\u00adomorphous space group P32. The structure of (III)D3-symmetric \u039b-Cu(bpy)32+ cations with CuII in an octa\u00adhedral CuN6 coordination environment. The Cu\u2014N distances are in agreement with those of the Cu(bpy)32+ cations in [Cu(bpy)3][PF6]2 it Fig.\u00a03.2O)2SnF6 and crystallizes in space group P2/n. The structure is composed of one-dimensional coordination chains propagating in the [101] direction that can be described as alternating Cu(bpy)(H2O)22+ cations (Cu site symmetry 2) and SnF62\u2212 anions catenated through bridging Cu\u2014F\u2014Sn linkages. The Sn4+ ion occupies a crystallographic inversion center. Intra\u00admolecular hydrogen bonding is present along the chains via O1\u2014H1A\u22efF2 and O1\u2014H1B\u22efF3 contacts ts Fig.\u00a04. The Cu\u20142(H2O)2+ and SiF62\u2212 groups are linked via O\u2014H\u22efF hydrogen bonding between the apical water mol\u00adecule and two SiF62\u2212 ions (Table\u00a012(H2O)2+ units participate in displaced heterochiral \u03c0\u2013\u03c0 stacking inter\u00adactions between the N1/C1\u2013C5 and N2/C6\u2013C10 rings with an inter\u00adplanar angle of 1.11\u2005(11)\u00b0, centroid\u2013centroid distance of 3.8774\u2005(12)\u2005\u00c5, and a slippage distance of 1.490\u2005\u00c5 to form \u0394up\u2013\u039bdown\u2013\u0394up\u2013\u039bdown and \u0394down\u2013\u039bup\u2013\u0394down\u2013\u039bup chains (up/down refers to the orientation of the Cu\u2014O bond vector in the +a or \u2013a direction). The water mol\u00adecules of hydration are involved in O\u2014H\u22efF hydrogen bonding inter\u00adactions with the SiF62\u2212 anion as well as O\u2014H\u22efO bonds with other water mol\u00adecules s Table\u00a01. The \u0394/\u039bes Fig.\u00a05.2(TaF6)2 complexes in (II)a + b or b \u2013 a directions, as shown in Fig.\u00a06c-axis direction, each chain is neighbored by a chain with the opposite chirality and same orientation on one side and a chain with the same chirality and opposite orientation on the other.The neutral Cu(bpy)32+ complexes participate in displaced \u03c0\u2013\u03c0 stacking inter\u00adactions propagating along the 32 screw axes with an inter\u00adplanar angle of 13.9\u2005(2)\u00b0, centroid\u2013centroid distance of 3.933\u2005(2)\u2005\u00c5 between adjacent N1/C1\u2013C5 and N5/C21\u2013C25 pyridine rings, and a horizontal shift distance of 1.970\u2005\u00c5. Each \u039b-Cu(bpy)32+ cation is surrounded by six TaF6\u2212 anions ns Fig.\u00a07.via parallel displaced \u03c0\u2013\u03c0 stacking inter\u00adactions ns Fig.\u00a08. One of et al., 20162(H2O)]2+ complexes and fluorinated inorganic anions: [Cu(bpy)2(H2O)][BF4]2 ][PF6]2 ][MF6] MF62\u2212 anions hydrogen bonded to the [Cu(bpy)2(H2O)]2+ complex are both hydrogen bonded to the same pair of [Cu(bpy)2)(H2O)]2+ complexes in the ETM case, whereas they are bound to two different complexes in the SiF62\u2212 case. Further, while the [Cu(bpy)2(H2O)][MF6] compounds display both face-to-face and displaced \u03c0\u2013\u03c0 stacking inter\u00adactions, (I)A survey of structures related to (I)et al., 2017a-axis direction. Additionally, these complexes participate in heterochiral \u03c0\u2013\u03c0 stacking inter\u00adactions.A search of the CSD for structures related to (II)A(bpy)3][PF6] 32+ complexes.Compound (III)2O)HfF6 (H2O)2MOxFx6\u2013 compounds , which display polar zigzag chains et al., 1993\u22121 to 150\u00b0C and held at 150\u00b0C for 24\u2005h. The autoclaves were allowed to cool to room temperature at a rate of 6\u00b0C h\u22121 and the solid products were recovered by vacuum filtration. Compound (I)3)2\u00b7H2O, 5\u2005mmol of 2,2\u2032-bi\u00adpyridine, 1.5\u2005mmol of (NH4)2SiF6 and 1ml of deionized H2O. Compound (II)2O5, 0.8\u2005ml HF(aq), and 0.3\u2005ml of deionized H2O. Compound (III)2O5, 1\u2005ml of HF(aq) and 0.1\u2005ml of deionized H2O. Compound (IV)3)2\u00b7H2O, 1.3\u2005mmol of 2,2\u2032-bi\u00adpyridine, 1.7\u2005mmol of (NH4)2SnF6 and 1\u2005ml of deionized H2O.The compounds reported here were synthesized by the hydro\u00adthermal pouch method (Harrison Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a03m1. The twinning occurs with a BASF of 0.5, suggesting that both the P31 and P32 configurations are present in equal proportions within the sample.The measured crystal of (III)10.1107/S2056989021000633/hb7957sup1.cifCrystal structure: contains datablock(s) global, II, III, IV, I. DOI: 2048914, 2048918, 2048917, 2048915CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Completion of the overall layer structure occurs through weak C\u2014H\u22efO, C\u2014H\u22ef\u03c0 (ring) and head-to-tail slipped \u03c0-stacking inter\u00adactions.The title mol\u00adecule is only a few degrees out of planarity except for the 2-hy\u00addroxy\u00adethyl substituent. In the crystal, O\u2014H\u22efN hydrogen bonds form stepped chains along the 12H13N3O2S, the benzo\u00adthia\u00adzine moiety is slightly non-planar, with the imidazolidine portion twisted only a few degrees out of the mean plane of the former. In the crystal, a layer structure parallel to the bc plane is formed by a combination of O\u2014HHydethy\u22efNThz hydrogen bonds and weak C\u2014HImdz\u22efOImdz and C\u2014HBnz\u22efOImdz inter\u00adactions, together with C\u2014HImdz\u22ef\u03c0(ring) and head-to-tail slipped \u03c0-stacking [centroid-to-centroid distances = 3.6507\u2005(7) and 3.6866\u2005(7)\u2005\u00c5] inter\u00adactions between thia\u00adzole rings. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (47.0%), H\u22efO/O\u22efH (16.9%), H\u22efC/C\u22efH (8.0%) and H\u22efS/S\u22efH (7.6%) inter\u00adactions. Hydrogen bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Computational chemistry indicates that in the crystal, C\u2014H\u22efN and C\u2014H\u22efO hydrogen-bond energies are 68.5 (for O\u2014HHydethy\u22efNThz), 60.1 (for C\u2014HBnz\u22efOImdz) and 41.8\u2005kJ\u2005mol\u22121 (for C\u2014HImdz\u22efOImdz). Density functional theory (DFT) optimized structures at the B3LYP/6\u2013311\u2005G level are compared with the experimentally determined mol\u00adecular structure in the solid state.In the title mol\u00adecule, C A plausible mechanism for the formationof the product, 1--3-(2-hy\u00addroxy\u00adeth\u00adyl)imid\u00adazolidin-2-one (I), is given in the reaction scheme.Compounds containing the benzo\u00adthia\u00adzole backbone have been studied extensively in both academic and industrial laboratories , carried out at the B3LYP/6-311G level, are compared with the experimentally determined mol\u00adecular structure in the solid state.I) and B (S1/N1/C1/C6/C7)]. A puckering analysis of the conformation of the imidazolidine ring C (N2/N3/C8-C10) gave the parameters Q(2) = 0.0767\u2005(14)\u2005\u00c5 and \u03c6(2) = 66.5\u2005(10)\u00b0. The conformation is described as an \u2018envelope on C9\u2032. This ring is almost coplanar with the thia\u00adzole ring B with a dihedral angle of 3.61\u2005(4)\u00b0 between their mean planes.In the title mol\u00adecule (I) Fig.\u00a01, the benHydethy\u22efNThz (Hydethy = hy\u00addroxy\u00adethyl and Thz = thia\u00adzole) hydrogen bonds and C\u2014HImdz\u22ef\u03c0(ring) inter\u00adactions (Table\u00a01Bnz\u22efOImdz (Bnz = benzene) inter\u00adactions. Both the layer formation and stacking are also assisted by head-to-tail slipped \u03c0-stacking inter\u00adactions and 3.6866\u2005(7)\u2005\u00c5, respectively; symmetry codes: (i) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01; (ii) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a0z\u00a0+\u00a0Cg2 is the centroid of ring B].In the crystal, O\u2014Hs Table\u00a01 form steon Fig.\u00a02. These as Table\u00a01. The lays Figs. 3 and 4 \u25b8 I, a Hirshfeld surface (HS) analysis has a symmetrical distribution of points with the edges at de + di = 2.40\u2005\u00c5. The presence of C\u2014H\u22ef\u03c0 inter\u00adactions is indicated by the characteristic wings with a spikes with the tips at de + di = 2.63\u2005\u00c5 in the fingerprint plot delineated into H\u22efC/C\u22efH contacts . The H\u22efS/S\u22efH contacts contribute 7.6% to the overall crystal packing and are seen in Fig.\u00a08e as widely scattered points with the tips at de + di = 3.03\u2005\u00c5. The pair of spikes in the fingerprint plot delineated into H\u22efN/N\u22efH contacts has a symmetrical distribution of points with the tips at de + di = 1.88\u2005\u00c5. The C\u22efC contacts have an arrow-shaped distribution of points with the tip at de = di = 1.70\u2005\u00c5. The N\u22efC/C\u22efN inter\u00adactions give rise to tiny wings with the tips at de + di = 3.41\u2005\u00c5. The O\u22efC/C\u22efO contacts give widely scattered points with the tips at de + di = 3.56\u2005\u00c5. Finally, the S\u22efC/C\u22efS and S\u22efN/N\u22efS inter\u00adactions, contributing 2.2% and 1.3% to the overall crystal packing give rise to tiny wings with the tips at de + di = 3.63\u2005\u00c5 and de + di = 3.63\u2005\u00c5, respectively.The overall two-dimensional fingerprint plot, Fig.\u00a08n Table\u00a02 is H\u22efH, ts Fig.\u00a08f, 5.3% ng Fig.\u00a08j and k dnorm plotted onto the surface are shown for the H\u22efH, H\u22efO/O\u22efH, H\u22efC/C\u22efH, H \u22ef S/S\u22efH, H\u22efN/N\u22efH and C\u22efC inter\u00adactions in Fig.\u00a09a--f, respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efO/O\u22efH and H\u22efC/C\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing is the sum of electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies were calculated to be \u221267.2 (Eele), \u221218.0 (Epol), \u221235.4 (Edis), 75.7 (Erep) and \u221268.5 (Etot) for O2\u2014H2A\u22efN1, \u221221.5 (Eele), \u22126.1 (Epol), \u221282.0 (Edis), 62.3 (Erep) and \u221260.1 (Etot) for C5\u2014H5\u22efO1 and \u22121.2 (Eele), \u22126.3 (Epol), \u221273.7 (Edis), 45.7 (Erep) and \u221241.8 (Etot) for C8\u2014H8B\u22efO1.The inter\u00admolecular inter\u00adaction energies were calculated by the CE\u2013B3LYP/6\u2013311G energy model available in I in the gas phase have been computed using the B3LYP functional level of theory and the 6-31G basis set \u00b0 while the corresponding dihedral angle in the others vary from 13.64\u00b0 in III to 0.61\u00b0 in V.A search of the Cambridge Structural Database . The mixture was stirred at 353\u2005K for 24\u2005h. The solid material was removed by filtration and the solvent evaporated in vacuo. The solid product was purified by recrystallization from ethanol to give colourless crystals (yield: 70%).To a mixture of 2-amino\u00adbenzo\u00adthia\u00adzole (2.22\u2005mmol), bis(2-chloro\u00adeth\u00adyl)amine (1.11\u2005mmol) and potassium carbonate (3.21\u2005mmol) in DMF (25\u2005mL) was added a catalytic amount of tetra-Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989020001723/jj2219sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020001723/jj2219Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020001723/jj2219Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020001723/jj2219Isup4.cmlSupporting information file. DOI: 1982595CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Correction to: BMC Public Health (2019) 19:1599https://doi.org/10.1186/s12889-019-7985-5It was highlighted that in the original article the dataIncorrect statementEven though the prevalence rate of hypertension in male (29.1%) was mildly lower than female (32.7%), our finding showed a significantly higher PRAL in the male (25.3\u2009\u00b1\u200920.2\u2009mEq/d) comparing to the female (19.4\u2009\u00b1\u200916.7\u2009mEq/d).Correct statementPrevalence rate of hypertension in male (32.7%) was mildly higher than female (29.1%) and a significantly higher PRAL in the male (25.3\u2009\u00b1\u200920.2\u2009mEq/d) was also identified comparing to the female (19.4\u2009\u00b1\u200916.7\u2009mEq/d)."} +{"text": "Febuxostat and ethanol mol\u00adecules are linked into an O\u2014H\u22efO and O\u2014H\u22efN bonded chain structure. 16H16N2O3S\u00b7C2H6O, (I), displays inter\u00admolecular O\u2014H\u22efO and O\u2014H\u22efN bonds in which the carboxyl group of the febuxostat mol\u00adecule and the hydroxyl group of the ethanol mol\u00adecule serve as hydrogen-bond donor sites. These inter\u00adactions result in a helical hydrogen-bonded chain structure. The title structure is isostructural with a previously reported methanol analogue.The title compound, 2-(3-cyano-4-iso\u00adbut\u00adoxyphen\u00adyl)-4-methyl-1,3-thia\u00adzole-5-car\u00adb\u00adoxy\u00adlic acid ethanol monosolvate, C The hy\u00addroxy group of the solvent additionally serves as a hydrogen-bond donor group for an O14\u2014H14\u22efO23 bond to a second febuxostat mol\u00adecule typically adopts a value close to 0\u00b0. However, an opposite geometry with \u03c4 values close to 180\u00b0 has been reported for the polymorphs Q and H1, a co-crystal with 4-amino\u00adbenzoic acid and a 2-(pyridin-2-yl\u00adamino)\u00adpyridinium salt.Table\u00a02The preparation of febuxostat was carried out according to the scheme in Fig.\u00a032, 10.0\u2005g) and hexa\u00admethyl\u00adene\u00adtetra\u00admine (5.86\u2005g) were added to tri\u00adfluoro\u00adacetic acid (100\u2005ml). The reaction mixture was heated to reflux under stirring for 40\u2005h, and tri\u00adfluoro\u00adacetic acid was distilled out. The obtained residue was cooled to 298\u2005K, water (200\u2005ml) was added slowly, and the slurry was stirred for 4\u2005h. After filtration, the product was washed and dried under vacuum to give 9.60\u2005g of 3.Ethyl 2-(4-hy\u00addroxy\u00adphen\u00adyl)-4-methyl-5-thia\u00adzole carboxyl\u00adate , potassium carbonate (332\u2005g) and isobutyl bromide (330\u2005g) were added to DMF (1.75 1). The reaction mixture was heated to 383\u00b13\u2005K and stirred for 4\u2005h. The reaction mixture was cooled to 298\u2005K, and water (0.50\u2005l) was added slowly. The slurry was stirred for 2\u2005h. After filtration, the product was washed and dried under vacuum to give 389\u2005g of 4. 1H NMR (CDCl3), 400\u2005MHz): \u03b4 = 1.079\u20131.101 , 1.366\u20131.413 , 2.185\u20132.230 , 2.769 , 3.914\u20133.935 , 4.316\u20134.387 , 7.045\u20137.074 , 8.188\u20138.225 , 8.353\u20138.361 .Ethyl 2-(3-formyl-4-hy\u00addroxy\u00adphen\u00adyl)-4-methyl-5-thia\u00adzole\u00adcarboxyl\u00adate , sodium formate (123\u2005g) and hydroxyl\u00adamine hydro\u00adchloride (84\u2005g) were successively added to formic acid (1.4\u2005l). The reaction mixture was heated to reflux and stirred for 5\u2005h to complete the reaction. The reaction solution was cooled to 298\u2005K, and water (2.8\u2005l) was slowly added. After stirring for approximately 1\u2005h, the slurry was filtered, the product was washed with water and dried under vacuum to give 321\u2005g of 5. 1H NMR (CDCl3), 400\u2005MHz): \u03b4 = 1.053\u20131.104 , 1.368\u20131.463 , 2.164\u20132.225 , 2.768 , 3.890\u20133.911 , 4.324\u20134.395 , 6.998\u20137.027 , 8 8.188\u20138.225 , 8.353\u20138.361 .Ethyl 2-(3-formyl-4-iso\u00adbut\u00adoxy\u00adphen\u00adyl)-4-methyl-5-thia\u00adzole\u00adcarboxyl\u00adate and potassium carbonate (200\u2005g) were successively added to a mixture of MeOH (7.5\u2005l) and water (250\u2005ml). To complete the reaction, the solution was heated to reflux for 3\u2005h under stirring. The clear solution was cooled, and vacuum was applied to distil out the solvent below 313\u2005K. Water (5\u2005l) was added to the residue. After stirring, EtOAc (2.5\u2005l) was added. The solution was stirred, and the layers were separated. The pH of the aqueous solution was adjusted to 2.5\u00b10.2 by adding diluted hydro\u00adchloric acid solution at 313\u2005K. After stirring for 1\u2005h, the slurry was filtered, and the product was washed with water and dried under vacuum to give 215\u2005g of 1.Ethyl 2-(3-cyano-4-iso\u00adbut\u00adoxy\u00adphen\u00adyl)-4-methyl-5-thia\u00adzole\u00adcarboxyl\u00adate (Febuxostat (1\u2005g) was dissolved in ethanol (10\u2005ml), which yielded a clear solution upon heating to 338\u2005K. After filtration, the solution was allowed to cool to room temperature, and the subsequent crystallization resulted in febuxostat ethanol solvate.Uiso parameters were set to 1.5Ueq(C) of the parent carbon atom. H atoms bonded to secondary CH2 (C\u2014H = 0.99\u2005\u00c5) or tertiary CH (C\u2014H = 0.99\u2005\u00c5) carbon atoms and H atoms bonded to C atoms in aromatic rings (C\u2014H = 0.95\u2005\u00c5) were positioned geometrically and refined with Uiso set to 1.2Ueq(C) of the parent carbon atom. H atoms in OH groups were identified in difference maps, refined with a distance restraint [O\u2014H = 0.84\u2005(2)\u2005\u00c5] and a free Uiso parameter. Two outliers I. DOI: 10.1107/S2056989020006076/fy2145Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020006076/fy2145Isup3.cmlSupporting information file. DOI: 2000973CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The structures of both compounds are dominated by hydrogen bonds involving water coordination ligands, with an underlying 3.6 \u2212 8H4N4O2)(H2O)4]n, (I), and [Sr(C8H4N4O2)(H2O)3]n, (II), from the one-pot hydrolysis transformation of benzoyl chloride and the in situ self-assembled [2\u00a0+\u00a03] cyclo\u00adaddition of nitrile are presented. These coordination compounds are prepared by reacting 4-cyano\u00adbenzoyl chloride with divalent alkaline-earth salts (BaCl2 and SrCl2) in aqueous solution under hydro\u00adthermal conditions. The mononuclear coordination compounds (I) and (II) show the same mode of coordination of the organic ligands. The cohesion of the crystalline structures is provided by hydrogen bonds and \u03c0-stacking inter\u00adactions, thus forming three-dimensional supra\u00admolecular networks. The two compounds have a three-dimensional -connected topology, and the structural differences between them is in the number of water mol\u00adecules around the alkaline earth metals. Having the same emission frequencies, the compounds exhibit photoluminescence properties with a downward absorption value from (I) to (II).Two alkaline-earth coordination compounds, [Ba(C In these two coordination compounds, the asymmetric unit comprises half of a crystallographically independent alkaline-earth metal ion, half of a deprotonated 4-(tetrrazol-5-yl)benzoate anion (ttzbenz), and two halves of water mol\u00adecules in compound (I)et al., 2009et al., 2007et al., 2009Compound (I)10 sphenocorona coordination geometry . In (I)2+ ion is eightfold coordinated, being surrounded by four bridging water mol\u00adecules and by four oxygen atoms from three symmetry-related ttzbenz ligands .The crystal structures of compounds (I)ds Fig.\u00a02, thus geAe2+ ion (Ae2+ = Ba2+ and Sr2+) range between 42.49\u2005(6) and 142.50\u2005(2)\u00b0 in compound (I)et al., 2010Ae2+ one-dimensional coordination polymers: Ba\u2014O = 2.647\u20133.179\u2005\u00c5, Sr\u2014O = 2.486\u20132.843\u2005\u00c5 in [C24H28N2O13Cl2CuSr]n and [C24H28N2O13Cl2CuBa]n 6(H2O)4]n, Ba\u2014O = 2.801\u2005(2)\u20133.6143\u2005(2)\u2005\u00c5 for M = Ni, Ba\u2014O = 2.797\u2005(2)\u20132.999\u2005(2)\u2005\u00c5 for M = Zn, and Ba\u2014O = 2.801\u2005(2)\u20133.004\u2005(2)\u2005\u00c5 for M = Co \u2005\u00c5 for compound (I)The ttzbenz ligand can adopt several coordination modes by involving the tetra\u00adzole ring ed Fig.\u00a04 with a cCg1) and the phenyl rings (centroid Cg2) with centroid\u2013centroid distances Cg1\u22efCg2 = 3.622\u2005(3)\u2005\u00c5 and Cg2\u22efCg2 = 3.897\u2005(3)\u2005\u00c5 \u2005\u00c5 Fig.\u00a04.et al., 2011c net with stoichiometry (3-C)2(6-C), which can be represented by the point symbol {43}2{46.66.83}. Thus the two structures consist of planar layers running parallel to (100) to obtain the underlying network. In such models, only metal centers and the centroids of organic ligands are considered as structural units (Alexandrov 0) Fig.\u00a05.et al., 2016et al., 2009et al., 2012et al., 2012et al., 2007et al., 2009et al., 2013et al., 2016et al., 2009et al., 2011et al., 2017et al., 2016et al., 2016et al., 2015et al., 2016N,N-di\u00admethyl\u00adacetamide benzoate in the Cambridge Structural Database for (I)2\u00b76H2O for (II)Colorless crystals suitable for X-ray diffraction were obtained by hydro\u00adthermal synthesis in an aqueous solution according to a literature procedure \u22121): 3300 \u03bd(O\u2014H)water, 3100 \u03bd(C\u2014H)Ph, 1435 \u03bdsym (C\u2014C), 1523 \u03bd(N\u2014N)ring, 1603 \u03bd(C\u2014N)ring, 628\u20131050 \u03b3,\u03b4 (tetra\u00adzole).FT\u2013IR of (I)\u22121): 3600 \u03bd(O\u2014H)water, 3200 \u03bd(C\u2014H)Ph, 1408 \u03bdsym (C\u2014C), 1530 \u03bd(N\u2014N)ring, 1585 \u03bd(C\u2014N)ring, 654\u20131009 \u03b3, \u03b4 (tetra\u00adzole) .FT\u2013IR of (II)The thermogravimetric analysis (TGA) was performed in the range 25\u2013600\u00b0C under air atmosphere at a flow rate of 5\u00b0C/min Fig.\u00a06. The pyr\u22121 was performed in the range 25\u2013600\u00b0C under an air atmosphere at a flow rate of 5\u00b0C min\u22121 Fig.\u00a06. The pyreX = 322\u2005nm) on an Agilent Cary Eclipse Fluorescence Spectrophotometer at room temperature. Excitation of the two compounds after dissolution in DMSO leads to similar fluorescence emission spectra. The emission maximum of (I)supporting information), probably corresponding to \u03c0* \u2192 \u03c0 or \u03c0*\u2192n electronic transition of the aromatic ring ttzbenz ligands Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989020006386/tx2021sup1.cifCrystal structure: contains datablock(s) TTZBENZ_AE, I, II. DOI: 10.1107/S2056989020006386/tx2021Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989020006386/tx2021IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989020006386/tx2021sup4.jpgInfrared spectra. DOI: Click here for additional data file.10.1107/S2056989020006386/tx2021sup5.jpgPhotoluminescent spectra. DOI: Click here for additional data file.10.1107/S2056989020006386/tx2021sup6.docTable S1. Comparative shape analysis of two polymers. DOI: 2003538, 2003537CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecule forms a centrosymmetric water-bridged hydrogen-bonding dimer with graph-set notation via two longer inter\u00admolecular hydrogen-bonding inter\u00adactions between the second hydrogen atom on the bridging water mol\u00adecule and both a pyridine nitro\u00adgen atom and carbonyl oxygen. The tri\u00adfluoro\u00admethyl groups extend out the faces of the sheet providing for F\u22efF and C\u2014H\u22efF contacts between the sheets, completing the three-dimensional packing.The title compound, [systematic name: 5-(tri\u00adfluoro\u00admeth\u00adyl)pyridine-2-carb\u00adoxy\u00adlic acid monohydrate], C 7H4F3NO2\u00b7H2O, is the acid hydrate of a pyridine with a carb\u00adoxy\u00adlic acid group and a tri\u00adfluoro\u00admethyl substituent situated para to one another on the aromatic ring. The mol\u00adecule forms a centrosymmetric water-bridged hydrogen-bonding dimer with graph-set notation R44 (12). Hydrogen-bonding distances of 2.5219\u2005(11) and 2.8213\u2005(11)\u2005\u00c5 are observed between the donor carb\u00adoxy\u00adlic acid and the bridging water acceptor, and the donor water and carbonyl oxygen acceptor, respectively. The dimers are further linked into a two-dimensional sheet via two longer inter\u00admolecular hydrogen-bonding inter\u00adactions between the second hydrogen atom on the bridging water mol\u00adecule and both a pyridine nitro\u00adgen atom and carbonyl oxygen with distances of 3.1769\u2005(11) and 2.8455\u2005(11)\u2005\u00c5, respectively. The tri\u00adfluoro\u00admethyl groups extend out the faces of the sheet providing for F\u22efF and C\u2014H\u22efF contacts between the sheets, completing the three-dimensional packing.The title compound [systematic name: 5-(tri\u00adfluoro\u00admeth\u00adyl)pyridine-2-carb\u00adoxy\u00adlic acid monohydrate], C The twoThe structure of 5-(tri\u00adfluoro\u00admeth\u00adyl)picolinic acid (I)W distance of 2.5219\u2005(11)\u2005\u00c5 characterizing the O1\u2014H1\u22efO1W hydrogen bond, while the water hydrogen atom H2W bonds to the carbonyl oxygen atom with an O1W\u22efO2i distance of 2.8213\u2005(11)\u2005\u00c5 charaterizing the O1W-\u2013H2W\u22efO2i hydrogen bond .The mol\u00adecular packing in the solid state can be characterized by the 5-(tri\u00adfluoro\u00admeth\u00adyl)picolinic acid (I)2) Fig.\u00a02. The carW, further bridge the dimer units together to form a pleated strip or tape motif that propagates along the crystallographic [010] direction and 2.8455\u2005(11)\u2005\u00c5, respectively, characterize the O1W\u2014H1W\u22efNii and O1W\u2014H1W\u22efO2ii hydrogen bonds NO3\u00b7H2O hydrogen bonds into thick two-dimensional sheets with the tri\u00adfluoro\u00admethyl\u00adaromatic groups extending to the faces of the sheets -2-pyridine\u00adcarb\u00adoxy\u00adlic acid has been used as a monoanionic ligand in several metal complexes, including with Co5-(Tri\u00adfluoro\u00admeth\u00adyl)-2-pyridine\u00adcarb\u00adoxy\u00adlic acid was purchased from Aldrich Chemical Company, USA, and was used as received.Uiso(H) = 1.2Ueq(C) of the aryl C-atoms. The position of the carb\u00adoxy\u00adlic acid and water hydrogen atoms were found in the difference map and refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a021H NMR : \u03b4 3.70 , 8.21 , 8.37 , 9.07 . 13C NMR : \u03b4 123.23 , 124.73 , 127.30 , 135.12 , 146.24 , 151.98 , 165.13 . 19F NMR : \u03b4 \u221261.35 . IR : 3469 , 3050 , 2849 (w), 2571 (w), 1961 (m), 1707 , 1606 (s), 1582 (s), 1493 (s), 1440 (s), 1392 (s), 1328 (m), 1290 (m), 1251 (s), 1163 (m), 1126 (s), 1075 (s), 1023 (s), 948 (s), 878 (m), 864 (s), 806 (s), 760 (s), 704 (s), 643 (s), 524 (s). GC\u2013MS (Agilent Technologies 7890A GC/5975C MS): +M = 191 amu, corresponding to the anhydrous form, 5-(tri\u00adfluoro\u00admeth\u00adyl)pyridine-2-carb\u00adoxy\u00adlic acid (I)10.1107/S2056989020013523/dj2015sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989020013523/dj2015Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020013523/dj2015Isup3.cmlSupporting information file. DOI: 2036131CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-018-22971-w, published online 16 March 2018Correction to: In Figure 4, the panels r and s were presented as s and r respectively. The correct Figure 4 appears below as Figure\u00a0In addition, this Article contains an error in the Methods section under subheading \u2018Collagen gel preparation\u2019 where\u201cRat tail type 1 collagen gels (Merck 08\u2013115) were prepared at 1\u2009mg/ml in 10\u2009\u00d7\u2009M199 (Life Technologies 11825\u2013015) and the pH was adjusted to 7.5 with 2.2% (w/v) NaHCO3. 10\u2009\u00d7\u2009M199 and NaHCO3 were both measured at 1/10 of the final volume.\u201dshould read:\u201cRat tail type 1 collagen gels (Merck 08\u2013115) were prepared at 1\u2009mg/ml in 10\u2009\u00d7\u2009M199 (Life Technologies 11825\u2013015) and the pH was adjusted with 2.2% (w/v) NaHCO3. 10\u2009\u00d7\u2009M199 and NaHCO3 were both measured at 1/10 of the final volume.\u201d"} +{"text": "The asymmetric unit of the title compound comprises three independent mol\u00adecules of similar geometry. The crystal structure is stabilized by inter\u00admolecular C\u2014H\u22efN and C\u2014H\u22efCl hydrogen bonds in addition to C\u2014Cl\u22ef\u03c0 inter\u00adactions. 16H14Cl3N3, comprises three mol\u00adecules of similar shape in the asymmetric unit. The crystal cohesion is ensured by inter\u00admolecular C\u2014H\u22efN and C\u2014H\u22efCl hydrogen bonds in addition to C\u2014Cl\u22ef\u03c0 inter\u00adactions. Hirshfeld surface analysis and two-dimensional fingerprint plots reveal that Cl\u22efH/H\u22efCl (33.6%), H\u22efH (27.9%) and C\u22efH/H\u22efC (17.6%) are the most important contributors towards the crystal packing.The title compound, C Nevertheless, they can control the reactivity of mol\u00adecules, the crystal packing, tautomerization and other properties -(4-chloro\u00adphen\u00adyl)diazen\u00adyl]ethen\u00adyl}-N,N-di\u00admethyl\u00adaniline, which features C\u2014H\u22efN, C\u2014H\u22ef\u03c0 and C\u2014Cl\u22efCl types of weak inter\u00admolecular inter\u00adactions.In a previous study we have attached resonance-assisted hydrogen-bonded synthons or chlorine atoms to dye mol\u00adecules, which leads to inter\u00admolecular weak inter\u00adactions for the resulting products with inter\u00adesting analytical and solvatochromic properties (Maharramov A (C1A\u2013C16A/N1A\u2013N3A/Cl1A\u2013Cl3A) and Mol-N1B (C1B\u2013C16B/N1B\u2013N3B/Cl1B\u2013Cl3B). The conformational differences between mol\u00adecules Mol-N1, Mol-N1A and Mol-N1B are highlighted in an overlay diagram shown in Fig.\u00a02A\u2013C6A and C8A\u2013C13A (mol\u00adecule Mol-N1A), and C1B\u2013C6B and C8B\u2013C13B (mol\u00adecule Mol-N1B)] of the 4-chloro\u00adphenyl and N,N-di\u00admethyl\u00adaniline groups are 69.94\u2005(10), 79.68\u2005(12) and 88.08\u2005(13)\u00b0, respectively. In mol\u00adecule Mol-N1, the N1\u2014N2\u2014C7\u2014C14, N2\u2014C7\u2014C14\u2014Cl2, N2\u2014C7\u2014C14\u2014Cl3 and C8\u2014C7\u2014C14\u2014Cl3 torsion angles are \u2212178.7\u2005(2), 3.1\u2005(3), \u2212176.21\u2005(16) and 4.1\u2005(3)\u00b0, respectively. The corres\u00adponding angles are 178.4\u2005(2), 3.8\u2005(3), \u2212175.1\u2005(2) and 2.5\u2005(3)\u00b0 for mol\u00adecule Mol-N1A, and \u2212175.0\u2005(2), 0.3\u2005(3), 179.71\u2005(18) and \u22120.1\u2005(4) for mol\u00adecule Mol-N1B.The asymmetric unit of the title compound Fig.\u00a01 containsIn the crystal, the mol\u00adecules are connected by inter\u00admolecular C\u2014H\u22efN and C\u2014H\u22efCl hydrogen bonds and C\u2014Cl\u22ef\u03c0 inter\u00adactions, which contribute to the overall packing, forming a three-dimensional network Table\u00a01.et al., 2007Crystal Explorer 17.5 . The shape-index of the Hirshfeld surface is a tool to visualize \u03c0\u2013\u03c0 stacking inter\u00adactions; Fig.\u00a04b clearly suggest that there are no \u03c0\u2013\u03c0 inter\u00adactions in the title compound. The red spots in Fig.\u00a04a correspond to the relatively strong C\u2014H\u22efN hydrogen-bonding inter\u00adactions in the crystal structure; in Mol-N1A it involves the N3A atoms of the N,N-di\u00admethyl\u00adaniline group as acceptors with the aromatic H2A donor atom of the chloro\u00adbenzene ring in Mol-N1 (C2\u2014H2A\u22efN3A).Hirshfeld surface analysis was used to investigate the presence of hydrogen bonds and inter\u00admolecular inter\u00adactions in the crystal structure. The Hirshfeld surfaces . Another significant reciprocal inter\u00adaction (H\u22efH) with a contribution of 27.9% is present as broad symmetrical spikes at diagonal axes de + di \u2243 2.2\u2005\u00c5 . The pair of characteristic wings in the fingerprint plot delineated into C\u22efH/H\u22efC contacts , have tips at de + di \u2243 2.80\u2005\u00c5. The Cl\u22efCl contacts, Fig.\u00a05e (5.7% contribution), have an arrow-shaped distribution of points with the tip at de = di = 3.50\u2005\u00c5.Two-dimensional fingerprint plots are presented in Fig.\u00a05ce Fig.\u00a05b. Anoth\u2005\u00c5 Fig.\u00a05c. The pviz. Cl\u22efC/C\u22efCl (5.4%), N\u22efH/H\u22efN (4.7%), C\u22efC (1.7%), Cl\u22efN/N\u22efCl (1.6%), N\u22efC/C\u22efN (1.0%) and N\u22efN (0.8%) contacts, show only small contributions and thus have a negligible effect on the packing.The other weak inter\u00admolecular inter\u00adactions, et al., 2016E)-1--2-phenyl\u00addiazene skeleton gave 25 hits, of which six closely resemble the title compound, viz. 1-(4-bromo\u00adphen\u00adyl)-2-diazene . Additional van der Waals inter\u00adactions consolidate the three-dimensional packing. In the crystal of HODQAV, mol\u00adecules are stacked in columns along [100] via weak C\u2014H\u22efCl hydrogen bonds and face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions. The crystal packing is further stabilized by short Cl\u22efCl contacts. In XIZREG, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds into zigzag chains running along [001]. The crystal packing is further stabilized by C\u2014Cl\u22ef\u03c0, C\u2014F\u22ef\u03c0 and N\u2014O\u22ef\u03c0 inter\u00adactions. In the crystal of LEQXIR, C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and Cl\u22efO contacts were found, and in LEQXOX, C\u2014H\u22efN and Cl\u22efCl contacts are observed.In the crystal structures of HONBOE and HONBUK, the aromatic rings form dihedral angles of 60.9\u2005(2) and 64.1\u2005(2)\u00b0, respectively. Mol\u00adecules are linked through weak et al., 2018E)-4-[(2-(4-chloro\u00adphen\u00adyl)hydrazineyl\u00adidene]meth\u00adyl)-N,N-di\u00admethyl\u00adaniline , tetra\u00admethyl\u00adethylenedi\u00adamine (TMEDA) , CuCl and CCl4 . After 1\u20133\u2005h , the reaction mixture was poured into a \u223c0.01 M solution of HCl and extracted with di\u00adchloro\u00admethane (3 \u00d7 20\u2005ml). The combined organic phase was washed with water (3 x 50\u2005ml), brine (30\u2005ml), dried over anhydrous Na2SO4 and concentrated in vacuo in a rotary evaporator. The residue was purified by column chromatography on silica gel using appropriate mixtures of hexane and di\u00adchloro\u00admethane (3/1\u20131/1) to give an orange solid. Yield: 72%; mp 408\u2005K. Analysis: calculated for C16H14Cl3N3: C 54.19, H 3.98, N 11.85; found: C 54.08, H 3.91, N 11.82%. 1H NMR \u03b4 3.05 , 6.79\u20137.79 . 13C NMR \u03b4 152.41, 151.45, 150.29, 137.26, 135.11, 131.08, 129.27, 124.50, 119.11, 111.48, 40.29. ESI\u2013MS: m/z: 355.48 [M + H]+.The title compound was synthesized according to a reported literature protocol = 0.93\u2005\u00c5, iUso(H) = 1.2Ueq(C) for aromatic H atoms, and 0.96\u2005\u00c5, Uiso(H) = 1.5Ueq(C) for methyl H atoms. Owing to poor agreement between observed and calculated intensities, five outliers I. DOI: 10.1107/S2056989020007549/wm5554Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020007549/wm5554Isup3.cmlSupporting information file. DOI: 2007970CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The phenol ring makes a dihedral angle of 44.77\u2005(3)\u00b0 with the benzene ring of the tri\u00adfluoro\u00admethyl group. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO inter\u00adactions, forming polymeric chain along the b-axis direction.The title compound, C 15H12F3NO, crystallizes with one mol\u00adecule in the asymmetric unit. The configuration of the C=N bond is E and there is an intra\u00admolecular O\u2014H\u22efN hydrogen bond present, forming an S(6) ring motif. The dihedral angle between the mean planes of the phenol and the 4-tri\u00adfluoro\u00admethyl\u00adphenyl rings is 44.77\u2005(3)\u00b0. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO inter\u00adactions, forming polymeric chains extending along the a-axis direction. The Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from C\u22efH/H\u22efC (29.2%), H\u22efH (28.6%), F\u22efH/H\u22efF (25.6%), O\u22efH/H\u22efO (5.7%) and F\u22efF (4.6%) inter\u00adactions. The density functional theory (DFT) optimized structure at the B3LYP/6-311\u2005G level is compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap. The crystal studied was refined as an inversion twin.The title compound, C These compounds show biological activities including anti\u00adbacterial, anti\u00adfungal, anti\u00adcancer and herbicidal activities carried out at the B3LYP/6-311\u2005G level are compared with the experimentally determined mol\u00adecular structure of (I)S(6) ring motif; this is a common feature in related imine\u2013phenol compounds. The imine group displays a C9\u2014C8\u2014N1\u2014C6 torsion angle of 170.1\u2005(4)\u00b0 while the mean plane of the phenol ring (C9\u2013C14) is inclined to that of the 4-tri\u00adfluoro\u00admethyl\u00adphenyl group (C1\u2013C6) by 44.77\u2005(3)\u00b0. The configuration of the C8=N1 bond is E. The C10\u2014O1 bond length [1.357\u2005(8)\u2005\u00c5 and 1.342\u2005\u00c5 ] indicates single-bond character a-axis direction on Fig.\u00a02. The cryet al., 2007CrystalExplorer17.5 and shape-index surfaces of (I)b) inter\u00adactions that involve C1\u2014H1A and the oxygen atom O1 of the phenol group de and di diagonal axes. The presence of C\u2014H\u22efO inter\u00adactions in the crystal is indicated by the pair of characteristic wings in the fingerprint plot delineated into C\u22efH/H\u22efC contacts (29.2% contributions to the Hirshfeld surface). In Fig.\u00a05c, the widely scattered points in the fingerprint plot are related to H\u22efH contacts, which make a contribution of 28.6% to the Hirshfeld surface. There are also F\u22efH/H\u22efF , O\u22efH/H\u22efO and F\u22efF contacts, with smaller contributions from N\u22efH/H.\u00b7N (2.4%), O\u22efC/C\u22efO (2.2%), F\u22efC/C\u22efF (0.8%) and O\u22efN/N\u22efO (0.2%) contacts.In order to visualize the role of weak inter\u00admolecular inter\u00adactions in the crystal, a Hirshfeld surface (HS) analysis using the standard B3LYP functional and the 6-311G basis-set calculations A = \u2212EHOMO), the ionization potential (I = \u2212ELUMO), the HOMO\u2013LUMO energy gap (\u0394E), the chemical hardness (\u03b7) and softness (S) and the lowest-unoccupied mol\u00adecular orbital (LUMO) are very important aspects as many electronic, optical and chemical reactivity properties of compounds can be predicted from these frontier mol\u00adecular orbitals -2-{[(3-chloro\u00adphen\u00adyl)imino]\u00admeth\u00adyl}-6-methyl\u00adphenol -2-[(2-hy\u00addroxy-5-meth\u00adoxy\u00adbenzyl\u00adidene)amino]\u00adbenzo\u00adnitrile -1-phenyl-N-imino}\u00admeth\u00adyl)phenolato]rhodium(III) chlorido\u00ad[2-({[4-(tri\u00adfluoro\u00admeth\u00adyl) phen\u00adyl]imino}\u00admeth\u00adyl)phen\u00adolato]iridium(III) sal\u00adicylaldiminato)titanium(IV) toluene solvate meth\u00adyl]amin\u00adato}copper(II) imino\u00admeth\u00adyl]phenolato}bis(tetra\u00adhydro\u00adfuran)\u00advanadium(III) imino\u00admeth\u00adyl]phenolato}(tetra\u00adhydro\u00adfuran)\u00advanadium(III) phen\u00adyl]-3-meth\u00adoxy\u00adsalicylaldimine -3-{[3-(tri\u00adfluoro\u00admeth\u00adyl)phenyl\u00adimino]\u00admeth\u00adyl}benz\u00adene-1,2-diol -5-(tri\u00adfluoro\u00admeth\u00adyl)aniline -2-meth\u00adyl-6-[3-(tri\u00adfluoro\u00admeth\u00adyl)phenyl\u00adimino\u00admeth\u00adyl]phenol -2-[(4-chloro\u00adphen\u00adyl)imino\u00admeth\u00adyl]-4-(tri\u00adfluoro\u00admeth\u00adoxy)phenol -4-methyl-2-[3-(tri\u00adfluoro\u00admeth\u00adyl)phenyl\u00adimino\u00admeth\u00adyl]phenol in ethanol (15\u2005mL) and 4-tri\u00adfluoro\u00admethyl\u00adphenyl\u00adamine in ethanol (15\u2005mL) and stirring the mixture for 8\u2005h under reflux. Single crystals suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution .Uiso(H) = 1.2\u20131.5Ueq(C). The hydrogen atom of the phenol group was located in a difference map and also included as a riding contributor with O\u2014H = 0.82\u2005\u00c5 and Uiso(H) = 1.5Ueq(O). During refinement, the twin transformation matrix , was used.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989020009615/mw2164sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020009615/mw2164Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020009615/mw2164Isup3.cmlSupporting information file. DOI: 2016363CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Pseudomonas putida and Bacillus subtilis. Notably, the more water\u2010soluble cIPTG derivative proved to be particularly suitable for light\u2010mediated gene expression in these alternative expression hosts.Photolabile protecting groups play a significant role in controlling biological functions and cellular processes in living cells and tissues, as light offers high spatiotemporal control, is non\u2010invasive as well as easily tuneable. In the recent past, photo\u2010responsive inducer molecules such as 6\u2010nitropiperonyl\u2010caged IPTG (NP\u2010cIPTG) have been used as optochemical tools for Lac repressor\u2010controlled microbial expression systems. To further expand the applicability of the versatile optochemical on\u2010switch, we have investigated whether the modulation of cIPTG water solubility can improve the light responsiveness of appropriate expression systems in bacteria. To this end, we developed two new cIPTG derivatives with different hydrophobicity and demonstrated both an easy applicability for the light\u2010mediated control of gene expression and a simple transferability of this optochemical toolbox to the biotechnologically relevant bacteria Extended toolbox: New photocaged IPTG derivatives with various hydrophobicities for light\u2010induced gene expression were synthesised and tested with three different expression host strains. The most water\u2010soluble compound showed superior performance, especially in Bacillus subtilis. A dean\u2010stark trap filled with molecular sieve (3\u2005\u00c5) was utilised for the constant removal of water. After the reaction was completed as indicated by TLC, it was washed with saturated NaHCO3 solution. The aqueous phase was then extracted with CH2Cl2 and the combined organic phase was dried with anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by flash column chromatography on SiO2 (petroleum ether/ethyl acetate 85\u2009:\u200915) to yield a yellow solid . Rf=0.25 (petroleum ether/ethyl acetate 80\u2009:\u200920) m.p. 62.1\u2009\u00b0C; 1H NMR : \u03b4=1.12 , 1.22 , 3.50 , 3.62 , 4.18 , 4.19 , 4.96 , 4.99 , 5.88 , 7.09 , 7.57\u2005ppm ; 13C NMR : \u03b4=14.0 (C\u201011 and C\u201011\u2032), 14.9 (C\u20102\u2032), 60.8 (C\u201010 or C\u201010\u2032), 60.9 (C\u201010 or C\u201010\u2032), 65.5 (C\u20108 or C\u20108\u2032), 65.6 (C\u20108 or C\u20108\u2032), 97.7 (C\u20107), 110.6 (C\u20103), 111.5 (C\u20106), 127.9 (C\u20101), 141.4 (C\u20102), 146.5 (C\u20104), 150.2 (C\u20105), 168.1 (C\u20109 or C\u20109\u2032), 168.1\u2005ppm (C\u20109 or C\u20109\u2032); IR (ATR\u2010film): \u1e7d=2981, 1755, 1692, 1581, 1526, 1446, 1346, 1291, 1196, 1176, 1080, 878, 796\u2005cm\u22121; HRMS (ESI): m/z calcd for C19H27NO10+: 447.1973 [M+NH4]+; found: 447.1972.Synthesis of BEC\u2010cIPTG (10\u2009a): To a solution of 4,5\u2010bis(ethoxycarbonylmethoxy)\u20102\u2010nitrobenzylaldehyde diethyl acetal (12) in dry CH2Cl2 (6\u2005mL) IPTG was added. After 5\u2005min p\u2010TSA was added to the suspension and it was stirred at room temperature for 20\u2005h. After the reaction was completed as indicated by TLC, a small amount of triethylamine was added and the reaction was concentrated under reduced pressure. The residue was purified by flash column chromatography on SiO2 (petroleum ether/ethyl acetate 50\u2009:\u200950 to 20\u2009:\u200980) to yield a white solid . Rf=0.35 (petroleum ether/ethyl acetate 20\u2009:\u200980); m.p. 104.5\u2009\u00b0C; [\u03b1]= \u221268 (c=1.0 in CHCl3); 1H NMR : \u03b4=1.31 , 1.35 , 1.36 , 2.56 , 3.25 , 3.52 , 3.64\u20133.70 , 4.08 , 4.24\u20134.31 , 4.41 , 4.77 , 4.82 , 6.21 , 7.35 , 7.54\u2005ppm ; 13C NMR : \u03b4=14.3 (C\u201011 or C\u201011\u2032), 14.3 (C\u201011 or C\u201011\u2032), 24.1 (CH3\u2010a or CH3\u2010b), 24.3 (CH3\u2010a or CH3\u2010b), 35.5 (SCH), 61.8 (C\u201010 or C\u201010\u2032), 61.9 (C\u201010 or C\u201010\u2032), 66.4 (C\u20108 or C\u20108\u2032), 66.6 (C\u20108 or C\u20108\u2032), 69.8 (C\u20106\u2032\u2032), 70.1 (C\u20105\u2032\u2032), 70.3 (C\u20103\u2032\u2032), 73.9 (C\u20102\u2032\u2032), 76.2 (C\u20104\u2032\u2032), 85.7 (C\u20101\u2032\u2032), 96.6 (C\u20107), 111.5 (C\u20103), 112.8 (C\u20106), 127.7 (C\u20101), 141.3 (C\u20102), 147.6 (C\u20104), 151.7 (C\u20105), 167.9 (C\u20109 or C\u20109\u2032), 167.9\u2005ppm (C\u20109 or C\u20109\u2032); IR (ATR\u2010film): \u1e7d=3478, 2967, 2916, 2866, 1747, 1520, 1287, 1176, 1097, 1077, 1027, 989\u2005cm\u22121; UV/Vis (MeOH): \u03bbmax (\u03f5)=298 nm (8006\u2005dm3 mol\u22121 cm\u22121); HRMS (ESI): m/z calcd for C24H37N2O13S: 593.2011 [M+NH4]+; found: 593.2011.Synthesis of BC\u2010cIPTG (10\u2009b): A solution of BEC\u2010cIPTG (10a) in MeOH (3.5\u2005mL) was cooled to 0\u2009\u00b0C and a 0.2\u2005m solution of LiOH (3.5\u2005mL) was added. The reaction mixture was stirred for 1\u2005h at room temperature. After the reaction was completed as indicated by TLC, the MeOH was evaporated under reduced pressure and the remaining solution was lyophilised overnight. The residue was suspended in THF, sonicated for 15\u2005min and filtrated. After washing with small amounts of cold THF a white solid was obtained. m.p. 190\u2009\u00b0C (decay); [\u03b1]=\u221292 (c=1.0 in H2O); 1H NMR : \u03b4=1.29 , 1.31 , 3.26 , 3.66 , 3.71\u20133.82 , 4.18 , 4.37 , 4.60 , 4.62 , 4.67 , 6.20 , 7.32 , 7.55\u2005ppm ; 13C NMR : \u03b4=22.9 (CH3\u2010a), 23.3 (CH3\u2010b), 35.0 (SCH), 67.3(C\u20108\u2032), 67.4 (C\u20108), 69.1 (C\u20102\u2032\u2032), 69.3 (C\u20106\u2032\u2032), 69.6 (C\u20105\u2032\u2032), 72.8 (C\u20103\u2032\u2032), 76.5 (C\u20104\u2032\u2032), 84.8 (C\u20101\u2032\u2032), 96.4 (C\u20107), 109.2 (C\u20103), 110.8 (C\u20106), 126.3 (C\u20101), 140.0 (C\u20102), 147.2 (C\u20104), 151.5 (C\u20105), 175.1 (C\u20109), 175.4\u2005ppm (C\u20109\u2032); IR (ATR\u2010film): \u1e7d=3124, 3043, 1605, 1522, 1398, 1335, 1277, 1077, 1047, 1024, 824\u2005cm\u22121; UV/Vis (H2O): \u03bbmax (\u03f5)=245 (5008), 342\u2005nm (3191\u2005dm3 mol\u20101 cm\u22121); HRMS (ESI): m/z calcd for C20H29N2O13S+: 537.1385 [M+NH4]+; found: 537.1382.Determination of purity by qNMR: The purity of the photocaged IPTG derivatives 10\u2009a, 10\u2009b and 1 was determined via quantitative NMR. 3,5\u2010bis(trifluoromethyl)bromobenzene was utilised as internal standard for 10\u2009a as well as 1 and (methanesulfonyl)methane for 10\u2009b. The spectra were measured at 20\u2009\u00b0C on a Bruker Avance/DRX 600 spectrometer with 64 scans each and 30\u2005\u03bcs relaxation time between each scan. The results in Table\u2005S3 are means of triplicate measurements.Solubility analysis: The solubility of 10\u2009a, 10\u2009b and 14 was determined photometrically at 25\u2009\u00b0C using a spectrophotometer Shimadzu UV\u20101800 (CPS\u2010240A). The absorbance of a serial dilution in degassed and deionised water was measured at the absorption maximum of the respective compound. A saturated solution was measured under the same conditions. The solubility was calculated using the Beer\u2010Lambert law.Hydrolytic stability: For the determination of the hydrolytic stability, a 1 mm solution of the respective compound in methanol or sodium phosphate buffer was stored in the dark at room temperature. Samples were removed after 0 and 24\u2005h and analysed by reversed\u2010phase HPLC.Quantification of uncaging half\u2010life times: A 1\u2005mm solution of each photocaged compound in methanol or sodium phosphate buffer was prepared. In a cuvette 1 mL of this solution was irradiated at room temperature using the LUMOS 43 (375\u2005nm) for a certain time period. The sample was then analysed by reverse phase HPLC Jasco HPLC system [column: Hyperclone 5\u2005\u03bc ODS (C18) 120 (Phenomenex)]. For each photocaged compound, the procedure was repeated for different irradiation times. The decrease of concentration was measured by an UV detector.Determination of uncaging quantum yields: The quantum yields of 1, 10\u2009a and 10\u2009b were determined by a relative method in comparison to the quantum yield of 2\u2010nitropiperonylacetate (NPA\u2010Ac), as this substrate shows a sufficient similarity to 1, 10\u2009a and 10\u2009b. The procedure was followed as previously described in literature .[Bacterial strains and plasmids: The E.\u2005coli strain DH5\u03b1E.\u2005coli strain S17\u20101E.\u2005coli strains, the P.\u2005putida strain KT2440B.\u2005subtilis strain DB430E.\u2005coli) or 30\u2009\u00b0C . Media were supplemented either with kanamycin (50\u2005\u03bcg\u2009mL\u22121), gentamicin (25\u2005\u03bcg\u2009mL\u22121), irgasan (25\u2005\u03bcg\u2009mL\u22121) or chloramphenicol (5\u2005\u03bcg\u2009mL\u22121), when appropriate.All bacterial strains and plasmids used in this study are listed in Table\u2005S1, Supporting Information.Plasmid construction: All recombinant DNA techniques were carried out as described by Sambrook et al.B.\u2005subtilis expression vector pHT01\u2010sfGFP, the sfGFP\u2010encoding gene was synthesised with flanking NdeI and HindIII restriction sites and subsequently cloned into pET\u201022(b) . The resulting vector pET\u201022(b)\u2010sfGFP was used as template for SLIC cloningsfgfp gene into the B.\u2005subtilis expression vector pHT01 using oligos 3\u20136 . The P.\u2005putida expression vector pVLT33\u2010GFPmut3 was constructed by restriction and ligation. To this end, the gfpmut3 gene was amplified with flanking EcoRI and XbaI restriction sites via PCR using oligos 1\u20132 (Table\u2005S1). Afterwards, the EcoRI/XbaI hydrolysed fragment was ligated into the likewise hydrolysed vector backbone pVLT33, resulting in the final expression vector pVLT33\u2010GFPmut3. Correct nucleotide sequences of all constructs were confirmed by Sanger sequencing (Eurofins Genomics).Cultivation conditions: All E.\u2005coli, P.\u2005putida and B.\u2005subtilis expression cultures were grown in 48\u2010well Flowerplates\u00ae in a BioLector microbioreactor system , inoculated with an optical density at 580\u2005nm of 0.05. During cultivation, the cell density was measured online through the scattered light intensity at 620\u2005nm. In addition, fluorescence of eYFP and GFP variants (GFPmut3 and sfGFP) were continuously determined using a 508/532\u2005nm and 488/520\u2005nm filter, respectively. cIPTG variants 10\u2009a, 10\u2009b or NP\u2010cIPTG (1) were added prior inoculation and expression of reporter genes was induced during the early logarithmic growth phase via UV\u2010A light exposure or by addition of equal amounts of conventional IPTG (14) after illumination.Determination of expression heterogeneity: For measurement of the expression heterogeneity, E.\u2005coli and B.\u2005subtilis cultures were analysed on the single\u2010cell level by flow cytometry regarding their fluorescence intensity and distribution. Expression cultures were grown as described above and were subsequently sampled as soon as they reached the late logarithmic growth phase (after 8\u2005h for E.\u2005coli and after 10\u2005h for B.\u2005subtilis). For this purpose, 40\u2005\u03bcL was taken out of the Flowerplate\u00ae cultures and added to 600\u2005\u03bcL PBS buffer (pH\u20057.4). Subsequently, the cells were harvested by centrifugation , adjusted to an optical density of 0.5 (OD580) in 100\u2005\u03bcL PBS buffer and then transferred into a 96\u2010well microtiter plate . Finally, these samples were analysed with a flow cytometer . The individual cellular fluorescence brightness was measured using a 488\u2010nm laser (15\u2009% intensity for E.\u2005coli and 5\u2009% for B.\u2005subtilis) for excitation and a 528/46\u2005nm bandpass filter for detection. To exclude cell debris and cell aggregates, the cells were also analysed regarding their size and granularity . FSC was measured using an FSC laser (nm) with 80\u2009% of the laser power for E.\u2005coli and 50\u2009% for B.\u2005subtilis and a 456/51\u2005nm bandpass filter for detection. For determination of SSC a nm\u2010light laser with 80\u2009% of the laser power for E.\u2005coli and 50\u2009% for B.\u2005subtilis (773/56\u2005nm bandpass filter) was used. Based on the scatter plots, bacterial cells were gated from irrelevant counts for fluorescence analysis. Flow cytometric data were evaluated with the CellStreamTM Analysis Software .The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "The crystal structure features both intra- and inter\u00admolecular C\u2014H\u22efO hydrogen bonds, as well as inter\u00admolecular C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions, leading to the formation of sheets parallel to the ac plane.The crystal structure of a pyrrolidine-substituted 11H15NO2S, features a sulfonamide group with S=O bond lengths of 1.4357\u2005(16) and 1.4349\u2005(16)\u2005\u00c5, an S\u2014N bond length of 1.625\u2005(2)\u2005\u00c5, and an S\u2014C bond length of 1.770\u2005(2)\u2005\u00c5. When viewing the mol\u00adecule down the S\u2014N bond, both N\u2014C bonds of the pyrrolidine ring are oriented gauche to the S\u2014C bond with torsion angles of \u221265.6\u2005(2)\u00b0 and 76.2\u2005(2)\u00b0. The crystal structure features both intra- and inter\u00admolecular C\u2014H\u22efO hydrogen bonds, as well as inter\u00admolecular C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions, leading to the formation of sheets parallel to the ac plane.The mol\u00adecular structure of the title compound, C Sulfonamides are of significant value in organic chemistry because of their therapeutic properties. These mol\u00adecules are referred to in the pharmaceutical industry as sulfa drugs. This class of drugs has been widely used in various pharmaceutical applications owing to their anti\u00adbacterial, anti\u00adviral, anti\u00admalarial, anti\u00adfungal, anti\u00adcancer, anti\u00addepressant, and other properties \u00b0 and C5\u2014S1\u2014N1\u2014C4 = 76.16\u2005(19)\u00b0. A conformational analysis of the five-membered pyrrolidine ring pucker gives a puckering amplitude (Q2) parameter of 0.352\u2005(3)\u2005\u00c5 and a \u03c62 parameter of 262.2\u2005(4)\u00b0. Consequently, this ring is in a half-chair conformation with a twist along the C2\u2014C3 bond. Lastly, an intra\u00admolecular C\u2014H\u22efO contact and a symmetry-derived ring . The \u03c0\u2013\u03c0 inter\u00adaction has a centroid-to-centroid distance of 3.8162\u2005(15)\u2005\u00c5 with a slippage of 1.307\u2005\u00c5. The result of these inter\u00adactions is the formation of sheets that lie in the ac plane to a stirring mixture of pyrrolidine and 10\u2005ml of tetra\u00adhydro\u00adfuran. This was followed by the dropwise addition of 0.59 M aqueous potassium carbonate and the mixture was stirred at room temperate for 6\u2005h. Upon acidification with 5 M HCl, a white precipitate was isolated by vacuum filtration to give the crude sulfonamide product. The crude product was dissolved in hot ethanol and filtered. The filtrate was transferred to a scintillation vial and crystallized upon standing for 24\u2005h to afford colorless crystals, filtered from the mother liquor .The title compound was prepared by the dropwise addition of Uiso(H) = 1.2Ueq(C) for methyl\u00adene groups and aromatic hydrogen atoms, and Uiso(H) = 1.5Ueq(C) for methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698902000208X/wm5540sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902000208X/wm5540Isup2.hklStructure factors: contains datablock(s) I. DOI: 1983920CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Phnl\u22efNPrmdn and N\u2014HPrmdn\u22efOPhnl (Phnl = phenol and Prmdn = perimidine) hydrogen bonds link the mol\u00adecules into infinite chains along the b-axis direction. C\u2014H\u22ef\u03c0 inter\u00adactions may further stabilize the crystal structure.The title compound consists of perimidin and meth\u00adoxy\u00adphenol units. In the crystal, O\u2014H 18H16N2O2, consists of perimidine and meth\u00adoxy\u00adphenol units, where the tricyclic perimidine unit contains a naphthalene ring system and a non-planar C4N2 ring adopting an envelope conformation with the NCN group hinged by 47.44\u2005(7)\u00b0 with respect to the best plane of the other five atoms. In the crystal, O\u2014HPhnl\u22efNPrmdn and N\u2014HPrmdn\u22efOPhnl (Phnl = phenol and Prmdn = perimidine) hydrogen bonds link the mol\u00adecules into infinite chains along the b-axis direction. Weak C\u2014H\u22ef\u03c0 inter\u00adactions may further stabilize the crystal structure. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (49.0%), H\u22efC/C\u22efH (35.8%) and H\u22efO/O\u22efH (12.0%) inter\u00adactions. Hydrogen bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Computational chemistry indicates that in the crystal, the O\u2014HPhnl\u22efNPrmdn and N\u2014HPrmdn\u22efOPhnl hydrogen-bond energies are 58.4 and 38.0\u2005kJ\u2005mol\u22121, respectively. Density functional theory (DFT) optimized structures at the B3LYP/ 6\u2013311\u2005G level are compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.The title compound, C We report herein the synthesis, the mol\u00adecular and crystal structures along with Hirshfeld surface analysis and computational calculations of the title compound, (I)Six-membered heterocyclic compounds carrying two nitro\u00adgen atoms have been widely studied , ring gave the parameters q2 = 0.3879\u2005(12)\u2005\u00c5, q3 = \u22120.2565\u2005(12)\u2005\u00c5, QT = 0.4650\u2005(13)\u2005\u00c5, \u03b82 = 123.47\u2005(15)\u00b0 and \u03c62 = 235.98\u2005(18)\u00b0]. The ring adopts an envelope conformation, where atom C1 is at the flap position and at a distance of 0.6454\u2005(12)\u2005\u00c5 from the best plane through the other five atoms. The C4N2 ring is hinged about the N\u22efN vector with the N1\u2014C1\u2014N2 plane being inclined by 47.44\u2005(7)\u00b0 to the best plane of the other five atoms (N1/N2/C9\u2013C11). In the meth\u00adoxy\u00adphenol moiety, the C8\u2014O2\u2014C4\u2014C5 and C8\u2014O2\u2014C4\u2014C3 torsion angles are \u22122.9\u2005(2)\u00b0 and 176.72\u2005(12)\u00b0, respectively. Rings A (C2\u2013C7), C (C10\u2013C15) and D (C9/C10/C15\u2013C18) are oriented at dihedral angles of A/C = 65.39\u2005(4)\u00b0, A/D = 69.63\u2005(4)\u00b0 and C/D = 4.31\u2005(3)\u00b0.The title compound, (I)ng Fig.\u00a01. A puckePhnl\u22efNPrmdn and N\u2014HPrmdn\u22efOPhnl (Phnl = phenol and Prmdn = perimidine) hydrogen bonds analysis have a symmetrical distribution of points with the tips at de + di = 3.03\u2005\u00c5. The H\u22efN/N\u22efH contacts have a distribution of points with the tips at de + di = 2.72\u2005\u00c5. The C\u22efC contacts have the tip at de = di = 3.37\u2005\u00c5. Finally, the O\u22efC/C\u22efO inter\u00adactions make only a 0.5% contribution to the overall crystal packing.The overall two-dimensional fingerprint plot, Fig.\u00a06dnorm plotted onto the surface are shown for the H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions in Fig.\u00a07a\u2013c, respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing is the sum of electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies were calculated as \u221237.5 (Eele), \u22127.8 (Epol), \u221252.0 (Edis), 52.4 (Erep) and \u221258.4 (Etot) of the mol\u00adecule is about 2.6162\u2005eV, and the frontier mol\u00adecular orbital energies, EHOMO and ELUMO are \u22123.1985 and \u22120.5823\u2005eV, respectively.The optimized structure of the title compound, (I)et al., 2011et al., 2013et al., 2013bet al., 2014et al., 2011et al., 2013bet al., 2013bet al., 2011et al., 2013et al., 2014Similar compounds of the perimidine derivative have also been reported and 1,8- di\u00adaminona\u00adphthalene (4\u2005mmol) in ether (30\u2005ml) under agitation at room temperature. Brown single crystals were obtained by the slow evaporation of the acetone solvent after 15 days (yield: 75%).The title compound, (I)C, H15A and H15B of the allyl moiety), 0.98\u2005\u00c5 (for methine H atom) and 0.97\u2005\u00c5 (for methyl\u00adene H atoms), and constrained to ride on their parent atoms, with Uiso(H) = 1.2Ueq(C). The hydroxyl H atom was placed in a calculated position with O\u2014H = 0.82\u2005\u00c5 and Uiso(H) = 1.5Ueq(O) while H atoms bonded to N atoms were refined independently with Uiso(H) = 1.2Ueq(N)Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989020004284/lh5952sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020004284/lh5952Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020004284/lh5952Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020004284/lh5952Isup4.cmlSupporting information file. DOI: 1976883CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the PDF version of this Article, the abbreviations for corresponding authors have been reversed\u201cCorrespondence and requests for materials should be addressed to Y.Z. (email: jinrong@bjmu.edu.cn) or R.J. (email: zhangyu007@bjmu.edu.cn)\u201dshould read:\u201cCorrespondence and requests for materials should be addressed to R.J. (email: jinrong@bjmu.edu.cn) or Y.Z. (email: zhangyu007@bjmu.edu.cn)This is correct in the HTML version of the ArticleIn the Materials and Methods section under the subheading \u201cThymic stromal cell isolation\u201d section,+), pDCs (CD11cint/hiCD45RA+), resident conventional DCs (CD11c+CD8\u03b1+CD172\u03b1\u2212), and migratory DCs (CD11c+CD8\u03b1\u2212CD172\u03b1\u2212)\u201d\u201cTo purify thymic DCs, cells were stained with fluorescence-labeled antibodies against CD11c, CD45RA, CD8\u03b1 and CD172\u03b1 and sorted on a BD FACS Aria II into the following fractions: total DCs (CD11cshould read:+), pDCs (CD11cint/hiCD45RA+), resident conventional DCs (CD11c+CD8\u03b1+CD172\u03b1\u2212), and migratory DCs (CD11c+CD8\u03b1\u2212CD172\u03b1+)\u201d\u201cTo purify thymic DCs, cells were stained with fluorescence-labeled antibodies against CD11c, CD45RA, CD8\u03b1 and CD172\u03b1 and sorted on a BD FACS Aria II into the following fractions: total DCs ,+ and CD172\uf061+ cDC was analyzed by flow cytometry following intracellular staining with anti-IL-27\u201d.\u201c(B) IL-27 protein expression in pDC, CD8\uf061Should read+ and CD172\u03b1+ cDC was analyzed by flow cytometry following intracellular staining with anti-IL-27\u201d.\u201c(B) IL-27 protein expression in pDC, CD8\u03b1Figure 6B and 6C were inverted. The correct Figure 6 appears below as"} +{"text": "After publication, an error was noticed in the \"Cell spreading is altered by RGD mutations\" section of the Results. The error occurred in the first paragraph, and the corrected text appears below.RGE and cFN\u0394RGD allows \u03b15\u03b21 and \u03b1v-class integrin\u2013mediated adhesion maturation and spreading, we plated pKO-\u03b1v, pKO-\u03b21, and pKO-\u03b1v/\u03b21 fibroblasts in serum replacement medium (SRM) on purified cFN variants and determined cell spreading as well as morphology and distribution of FAs and pKO-\u03b1v/\u03b21 fibroblasts (79.6 \u00b1 7.9%) spread on cFNRGE; and a neglectable percentage of pKO-\u03b21 fibroblasts spread on cFNRGE. On cFN\u0394RGD, the numbers of spread pKO-\u03b1v and pKO-\u03b1v/\u03b21 fibroblasts dropped to 22.2 \u00b1 15% and 10.4 \u00b1 7.0%, respectively, and of pKO-\u03b21 fibroblasts to <1% , similar to pKO-\u03b1v (583 \u00b1 42 \u00b5m2) and pKO-\u03b1v/\u03b21 fibroblasts (514 \u00b1 49 \u00b5m2) on cFNRGE. 2 (P < 0.05 compared to pKO-\u03b1v cells) on cFNRGE. In RGE cells from P < 0.001 to P < 0.05.The few adherent pKO-\u03b21 fibroblasts covered a reduced area of 419 \u00b1 36 \u00b5m On cFN\u0394RGD, the adherent pKO-\u03b1v, pKO-\u03b21, and pKO-\u03b1v/\u03b21 fibroblasts covered significantly smaller spreading areas when compared with the same cell lines seeded on cFNRGD (\"To test whether full-length cFNn of FAs . At 60 ms to <1% . Moreoven cFNRGD .\"The P values in"} +{"text": "II ion lies on a crystallographic twofold rotation axis and is coordinated by four P atoms from two -1,2-bis\u00ad(bi\u00adnaphthyl\u00adphospho\u00adnito)ethane (BPE) ligands and two Cl ligands in a distorted cis-FeCl2P4 octa\u00adhedral coordination geometry. Weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions occur in the crystal.In the title compound, the Fe 2,11.03,8.018,23]tricosa-1(15),2(11),3(8),4,6,9,16,18(23),19,21-deca\u00aden-13-yl]ethane}\u00addichlorido\u00adiron(II) di\u00adchloro\u00admethane disolvate), [FeCl2(C42H28O4P2)2]\u00b72CH2Cl2, the FeII ion lies on a crystallographic twofold rotation axis and is coordinated by four P atoms from two -1,2-bis\u00ad(bi\u00adnaphthyl\u00adphospho\u00adn\u00adito)ethane (BPE) ligands and two Cl ligands in a distorted cis-FeCl2P4 octa\u00adhedral coordination geometry. In the crystal, weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions link the mol\u00adecules into layers lying parallel to (001). A weak intra\u00admolecular C\u2014H\u22efO hydrogen bond is also observed. The asymmetric unit contains one CH2Cl2 solvent mol\u00adecule, which is disordered over two sets of site with refined occupancies in the ratio 0.700\u2005(6):0.300\u2005(6).In the title compound (systematic name: bis\u00ad{1,2-bis[12,14-dioxa-13-phospha\u00adpenta\u00adcyclo\u00ad[13.8.0.0 R,R)- or -1,2-bis\u00ad(bi\u00adnaphthyl\u00adphospho\u00adnito)ethane prepared from either (R)- or (S)-1,1\u2032-bi(2-naphthol) has been used extensively in asymmetric catalysis, as has the related ligand (R) or (S)-2,2\u2032-bis\u00ad(di\u00adphenyl\u00adphosphino)-1,1\u2032-binaphthyl . For example, the BINAP ligand has been coordinated to ruthenium and used for the asymmetric hydrogenation of ketones 2]. The BPE ligand and similar bidentate and monodentate phospho\u00adnite ligands have been coordinated to rhodium and iridium and used for asymmetric alkene and quinoline hydrogenation reactions, respectively 2, which crystallized as a di\u00adchloro\u00admethane solvate.As an extension of these studies, we now describe the synthesis and crystal structure of the iron(II) complex FeClII ion lies on a crystallographic twofold rotation axis and is coordinated by four P atoms from two BPE ligands and two Cl ligands in a distorted cis-FeCl2P4 octa\u00adhedral coordination geometry. The largest distortion from ideal coordination geometry is the P2\u2014Fe\u2014P2i angle of 108.49\u2005(7)\u00b0 are the same, with values of 54.5\u2005(2)\u00b0. A weak intra\u00admolecular C\u2014H\u22efO hydrogen bond is observed :0.300\u2005(6).The mol\u00adecular structure of the title compound is shown in Fig.\u00a01d Table\u00a02. The asyCg2\u22efCg2 of 4.171\u2005(5)\u2005\u00c5 (where Cg2 is the centroid of the C4\u2013C9 benzene ring) may be a very weak \u03c0-stacking inter\u00adaction.In the crystal, weak C\u2014H\u22efO hydrogen bonds link mol\u00adecules into sheets parallel to (001) Table\u00a02. Within et al., 20162(P2)2 (P2 = a diphosphine) with P\u2014C bonds reported. The majority, 33 complexes, crystallize with the chloride ions trans to each other, while there are three examples where the chloride ions are cis, as in the title complex. The complex trans-FeCl2ethyl\u00adene)2, for example, crystallizes with the chloride ions trans 2 and 2.3423\u2005(11)\u2005\u00c5 in the title complex.A search of the Cambridge Structural Database -BINOL was combined with -BPE in 10\u2005ml THF and stirred in a 20\u2005ml dram vial for 24\u2005h. The THF was vacuumed off to yield a brown powder: 31P{1H} NMR : 257.72 ppm, singlet.The ligand was synthesized according to a literature procedure using = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a032Cl2 solvent mol\u00adecule was refined without restraints while the minor component was restrained to have similar geometry and anisotropic displacement parameters to the major component using the SAME and SADI instructions in SHELXL I. DOI: 10.1107/S2056989020011160/hb7939Isup2.hklStructure factors: contains datablock(s) I. DOI: 2023248CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "An intra\u00admolecular short S\u22efO contact [3.215\u2005(2)\u2005\u00c5] is observed. The crystal packing features C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, the dihedral angle between the fused pyrazole and pyridine rings is 1.76\u2005(7)\u00b0. The benzene and meth\u00adoxy phenyl rings make dihedral angles of 44.8\u2005(5) and 63.86\u2005(5)\u00b0, respectively, with the pyrazolo\u00ad pyridine moiety. An intra\u00admolecular short S\u22efO contact [3.215\u2005(2)\u2005\u00c5] is observed. The crystal packing features C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, C The thio\u00admethyl (\u2013SCH3) group fused to the pyrazolo\u00adpyridine unit exhibits a (+)anti-periplanar conformation [C8\u2014S1\u2014C7\u2014C6 = 175.47\u2005(15)\u00b0]. The torsion angles involving the \u2013SCH3 group differ from those for NUDWOB group. The \u2013COOC2H5 group attached to the pyrazolo\u00adpyridine moiety has a (+)anti-periplanar conformation [N3\u2014C7\u2014C6\u2014C9 = 176.44\u2005(16)\u00b0]. Further, the methyl group attached to the pyrazole subunit is (+)anti-periplanar [C1\u2014N2\u2014N1\u2014C4 = 178.78\u2005(17)\u00b0] and it is attached to the pyridine ring showing a (+)syn-periplanar conformation [C1\u2014N2\u2014C2\u2014N3 = 1.3\u2005(3)\u00b0]. The fused pyrazole and pyridine rings in the title compound are not exactly planar, as in NUDWOB \u00b0 and that between the meth\u00adoxy\u00adphenyl and pyrazolo\u00adpyridine rings is 63.86\u2005(5)\u00b0.In the title compound Fig.\u00a01, the pheCg and C23\u2014H23\u22efCg) inter\u00adactions = 3.291\u2005(2)\u2005\u00c5] may also exist noted by Thomas et al. methanone; Ravi et al., 2017a]pyridine; Wu et al., 2016H-pyrazolo\u00adpyridine-5-carboxyl\u00adate; Rao et al., 20202CH3 substituent in the title compound are comparable with those reported for FIZLEI and NUDWOB. The bond distances for the thio\u00admethyl and aryl moieties in the title compound are comparable with those of NUDWOB. Moreover, the bond lengths in the pyrazolopyridine unit of the title compound are comparable with those in NUDWOB, FIZLEI, ALAFID, DAWKAQ and NADPIU. The pyrazolo\u00adpyridine moiety (N1\u2013N3/C2\u2013C4/C5\u2013C7) of the title compound is approximately planar, as is also observed for NUDWOB, FIZLEI, ALAFID, DAWKAQ and NADPIU. In the title compound, a short intra\u00admolecular S\u22efO contact of 3.215\u2005(2)\u2005\u00c5 occurs, but this is not observed in FIZLEI, ALAFID, DAWKAQ, NADPIU and NUDWOB. Moreover, the inter\u00adaction distance [3.291\u2005(2)\u2005\u00c5] of the short inter\u00admolecular C\u22efO contact in the title compound is comparable with the C\u22efO inter\u00adaction [3.424\u2005(2)\u2005\u00c5] in the structure of NUDWOB. Furthermore, as in the title compound, C\u2014H\u22ef\u03c0 inter\u00adactions are observed in the crystal structures of ALAFID, DAWKAQ and NADPIU. The inter\u00adaction distance of these related structures ranges from 2.89 to 3.23\u2005\u00c5, comparable with the C\u2014H\u22ef\u03c0 inter\u00adactions observed in the title compound.A similarity search of the Cambridge Structural Database and ethyl 2-(4-meth\u00adoxy\u00adbenzo\u00adyl)-3,3-bis\u00ad(methyl\u00adthio)\u00adacrylate in toluene (3\u2005mL), a catalytic amount of TFA (tri\u00adfluoro\u00adacetic acid) 30\u2005mol % in toluene (3\u2005mL) was added under an N2 atmosphere. The reaction mixture was refluxed for 24\u2005h in an oil bath, the progress of the reaction being monitored by TLC using a mixture of hexane and ethyl acetate (9.9:0.1). After completion of the reaction, the mixture was loaded on a silica gel column and eluted with increasing amounts of ethyl acetate in hexa\u00adnes (1% to 5%) to obtain 186\u2005mg (yield = 75%) of ethyl 6-(4-meth\u00adoxy\u00adphen\u00adyl)-1-methyl-4-(methyl\u00adthio)-3-phenyl-1H-pyrazolo\u00adpyridine-5-carboxyl\u00adate as a colourless crystalline solid; m.p. 406\u2005K; Rf = 0.3\u2005cm (hexa\u00adne: ethyl acetate 9.9:0.1). A sample suitable for single-crystal X-ray analysis was obtained by recrystallization from 2\u2005mL of dry methanol.To a solution of 1-methyl-3-phenyl-1Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020008841/dx2029sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020008841/dx2029Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020008841/dx2029Isup3.cmlSupporting information file. DOI: 2005976CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Acetamino\u00adphen assists as a co-former in the crystallization of hexa\u00admethyl\u00adene\u00adtetra\u00admine carboacetamino\u00adphenborane, which may otherwise be challenging to form a crystal itself. 15H22BN5O3\u00b7C8H9NO2. In the first of these mol\u00adecules, both the borate-ester and acetyl\u00adamino groups are considerably twisted away from the plane of the inter\u00advening benzene ring . The extended structure of this co-crystal features N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, which link the components into (100) sheets and weak C\u2014H\u22efO hydrogen bonds help to consolidate the structure.Hexa\u00admethyl\u00adene\u00adtetra\u00admine carboacetamino\u00adphenborane, a mol\u00adecule with two pharmacophores attached to a central carb\u00adoxy\u00adborate moiety, was synthesized and crystals were grown with an acetamino\u00adphen co-crystal former to result in the title 1:1 co-crystal [hexa\u00admethyl\u00adene\u00adtetra\u00admine 4-acetamido\u00adphenyl 2-boranyl\u00adacetate\u20134-acet\u00adamido\u00adphenol (1/1)], C This type of bond has been previously reported in the acetamino\u00adphen co-crystal with citric acid of this hydrogen bond is similar to the N\u2014H\u22efO\u2014H bond (2.09\u2005\u00c5) from the known acetamino\u00adphen crystal form II is identical in length to that of acetamino\u00adphen crystal form II (1.85\u2005\u00c5). Several weak C\u2014H\u22efO hydrogen bonds may help to consolidate the structure.During crystallization, the new difunctionalized mol\u00adecule, CORCB-1-APAP, forms a co-crystal with acetamino\u00adphen at a 1:1 ratio with hydrogen-bonding inter\u00adactions Table\u00a01 between et al., 1998A mol\u00adecular packing projection of (I)et al., 1980et al., 2002et al., 2002et al., 2017et al., 2009et al., 2011et al., 2007et al., 2002The crystal structures of amine carb\u00adoxy\u00adborane have been reported as dimers involves several steps using tri\u00admethyl\u00adamine carb\u00adoxy\u00adborane (CORCB-3) as the starting material. Tri\u00admethyl\u00adamine carb\u00adoxy\u00adborane, synthesized by the previously reported method = 1.2Ueq or 1.2Ueq(Cmeth\u00adyl)].Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020015327/hb7947sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020015327/hb7947Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989020015327/hb7947sup4.pdfCheckCIF. DOI: 1828957CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules of the title compound, a potential ligand containing two 8-quinolinol and one 2-pyridine units, are linked by inter\u00admolecular O\u2014H\u22efN and O\u2014H\u22efO quadruple hydrogen bonds, forming an inversion dimer with two 24H15Cl2N3O2, one quinoline ring system is essentially planar and the other is slightly bent. An intra\u00admolecular O\u2014H\u22efN hydrogen bond involving the hy\u00addroxy group and a pyridine N atom forms an S(5) ring motif. In the crystal, two mol\u00adecules are associated into an inversion dimer with two R22(7) ring motifs through inter\u00admolecular O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds. The dimers are further linked by an inter\u00admolecular C\u2014H\u22efO hydrogen bond and four C\u2014H\u22ef\u03c0 inter\u00adactions, forming a two-dimensional network parallel to (001).In the title compound, C The torsion angles C8\u2014C16\u2014C17\u2014C18, C26\u2014C18\u2014C17\u2014C27, and C28\u2014C27\u2014C17\u2014C16 are \u221288.6\u2005(2), \u2212101.2\u2005(2) and \u221287.8\u2005(2)\u00b0, respectively.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01fs Fig.\u00a01. The arri and O4\u2014H4\u22efN5i; symmetry code: (i) \u2013x\u00a0+\u00a01, \u2013y\u00a0+\u00a02, \u2013z\u00a0+\u00a02], forming an inversion dimer with two et al., 1999Cg1iii and C24\u2014H24\u22efCg3v; Cg1 is the centroid of the C18\u2013C21/C25/C26 ring and Cg3 is the centroid of the N7/C27\u2013C31 ring; symmetry codes: (iii) x\u00a0+\u00a01, y\u00a0+\u00a01, z; (v) \u2013x, \u2013y\u00a0+\u00a01, \u2013z\u00a0+\u00a02], forming a ribbon structure along [110] x \u2013 1, y, z] and a C\u2014H\u22ef\u03c0 inter\u00adaction . The chains are linked by two C\u2014H\u22ef\u03c0 inter\u00adactions , generating a two-dimensional network parallel to (001) Fig.\u00a04.et al., 2016et al., 19992 methane and ethanol (6\u2005mL) were placed in a 15\u2005mL capped pressure tube. It was heated at 353\u2005K for 96\u2005h. The generated pale-white precipitate was filtered to give a pale-white solid . Single crystals of title compound suitable for X-ray diffraction were grown by slow evaporation of a solution in CHCl3/n-hexane at ambient temperature. 1H NMR \u03b4 = 6.63 , 7.21 , 7.33 , 7.52 , 7.52 , 7.67 , 8.48 , 8.64 , 8.81 , 8.84 .The title compound was prepared by a modification of the reported KUiso(H) = 1.2Ueq (C). One outlier global, I. DOI: 10.1107/S2056989020009317/is5545Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020009317/is5545Isup3.cmlSupporting information file. DOI: 2014831CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "These are connected in three-dimensions by benzene-C\u2014H\u22efO(DTBA), DTBA-C=O\u22ef\u03c0(benzene) and benzene-C\u2014H\u22ef\u03c0(benzene) inter\u00adactions.In the title 1:1 solvate, 2,2\u2032-di\u00adthiodi\u00adbenzoic acid (DTBA):di\u00admethyl\u00adformamide (DMF), the DTBA mol\u00adecule is twisted [C\u2014S\u2014S\u2014C = \u221288.57\u2005(6)\u00b0]. Four-mol\u00adecule aggregates are formed in the crystal 14H10O4S2\u00b7C3H7NO, features a twisted mol\u00adecule of 2,2\u2032-di\u00adthiodi\u00adbenzoic acid (DTBA), with the central C\u2014S\u2014S\u2014C torsion angle being \u221288.57\u2005(6)\u00b0, and a mol\u00adecule of di\u00admethyl\u00adformamide (DMF). The carb\u00adoxy\u00adlic acid groups are, respectively, close to co-planar and twisted with respect to the benzene rings to which they are connected as seen in the CO2/C6 torsion angles of 1.03\u2005(19) and 7.4\u2005(2)\u00b0. Intra\u00admolecular, hypervalent S\u2190O inter\u00adactions are noted [S\u22efO = 2.6140\u2005(9) and 2.6827\u2005(9)\u2005\u00c5]. In the crystal, four-mol\u00adecule aggregates are formed via DTBA-O\u2014H\u22efO(DMF) and DTBA-O\u2014H\u22efO(DTBA) hydrogen bonding, the latter via an eight-membered {\u22efOHCO}2 homosynthon. These are linked into supra\u00admolecular layers parallel to (011) via benzene-C\u2014H\u22efO(DTBA) and DTBA-C=O\u22ef\u03c0(benzene) inter\u00adactions, with the connections between these, giving rise to a three-dimensional architecture, being of the type benzene-C\u2014H\u22ef\u03c0(benzene). An analysis of the calculated Hirshfeld surfaces indicates, in addition to the aforementioned inter\u00admolecular contacts, the presence of stabilizing inter\u00adactions between a benzene ring and a quasi-\u03c0-system defined by O\u2014H\u22efO hydrogen bonds between a DTBA dimer, i.e. the eight-membered {\u22efOCOH}2 ring system, and between a benzene ring and a quasi-\u03c0(OCOH\u22efOCH) system arising from the DTBA-O\u2014H\u22efO(DMF) hydrogen bond. The inter-centroid separations are 3.65 and 3.49\u2005\u00c5, respectively.The title 1:1 solvate, C In a recent study, it was found the anti\u00adcipated {\u22efHOCO}2 synthon was not always formed but was usurped by a DTBA-O\u2014H\u22efO(DMF) hydrogen bond for one of the carb\u00adoxy\u00adlic acids, i.e. in the 1:1:1 co-crystal solvate DTBA:2-ClBA:DMF solution also gave the DTBA:2DMF solvate solution yielded the mono-solvate, i.e. the title compound DTBA:DMF, (I)Co-crystal formation with 2-mercapto\u00adbenzoic acid (2-MBA) is fraught as during crystallization, this is usually oxidized to 2,2\u2032-di\u00adthiodi\u00adbenzoic acid (DTBA) solution indicating the acid oxidized to DTBA during crystallization. The observed disparity in the C\u2014O bond lengths in the carb\u00adoxy\u00adlic acid residues confirms the location of the acidic H atoms on the O1 and O3 atoms, respectively. A characteristic twisted conformation is evidenced in the C3\u2014S1\u2014S2\u2014C8 torsion angle of \u221288.57\u2005(6)\u00b0. The dihedral angle between the benzene rings is 87.71\u2005(3)\u00b0, consistent with an orthogonal disposition. The C1-carb\u00adoxy\u00adlic acid group is almost co-planar with the (C2\u2013C7) benzene ring to which it is connected with the dihedral angle between the least-squares planes being 1.03\u2005(19)\u00b0. By contrast, a small twist is noted for the C14-carb\u00adoxy\u00adlic acid residue where the comparable dihedral angle is 7.4\u2005(2)\u00b0. Intra\u00admolecular hypervalent S\u2190O inter\u00adactions via an eight-membered {\u22efOHCO}2 homosynthon. The result is the four-mol\u00adecule aggregate shown in Fig.\u00a02a). For the DTBA\u22efDMF inter\u00adaction, further stabilization is realized through a DMF-C15\u2014H\u22efO2(carbon\u00adyl) contact, Table\u00a01via benzene-C7\u2014H\u22efO(hydrox\u00adyl) inter\u00adactions occurring between centrosymmetrically related mol\u00adecules. The chains are connected by parallel C=O\u22ef\u03c0(benzene) inter\u00adactions as detailed in Fig.\u00a02b) and Table\u00a01c), with connections between them leading to a three-dimensional architecture being benzene-C11\u2014H\u22ef\u03c0(benzene), Fig.\u00a02d).The key feature of the supra\u00admolecular aggregation in the crystal of (I)Mercury Crystal Explorer 17 and hy\u00addroxy-O3\u2014H3O\u22efO4(carbon\u00adyl) hydrogen bonds with the corresponding dnorm contact distances being 1.62 and 1.64\u2005\u00c5, respectively, i.e. significantly shorter by almost 1\u2005\u00c5 compared to the sum of the van der Waals radii of 2.61\u2005\u00c5 , Table\u00a02To better comprehend the supra\u00admolecular features of (I)\u03c0\u2013\u03c0 inter\u00adaction formed between the C8\u2013C13 benzene ring and a quasi-\u03c0-system defined by O3\u2014H3O\u22efO4 hydrogen bonds between a DTBA dimer, i.e. the eight-membered {\u22efO4\u2013C14\u2013O3\u2013H3O}2 ring system. A similar observation is also noted for the C1\u22efC15 contact which is encapsulated within an apparent \u03c0(C2\u2013C7)\u22efquasi-\u03c0(O2\u2013C1\u2013O1\u2013H1O\u22efO5\u2013C15\u2013H15) inter\u00adaction. The separation between the ring centroids of the aforementioned \u03c0\u2013\u03c0 contacts are 3.65 and 3.49\u2005\u00c5, respectively. The stacking arrangement between the relevant aromatic and quasi-aromatic rings is supported by shape complementarity as revealed by the concave (red) and convex (blue) regions in the shape index, Fig.\u00a05a)\u2013(d), as well as curvedness mappings, Fig.\u00a05e) and (f), obtained through the Hirshfeld surface analysis.Of particular inter\u00adest among all close contacts present in (I)d,p) approach to verify the nature of the contacts present in (I)O\u22efO5 followed by H3O\u22efO4 inter\u00adactions which is consistent with their corresponding short contact distances. By contrast, for the H\u22efC and C\u22efO inter\u00adactions relatively smaller charge disparity is noted indicating weaker attractions between the participating atoms,. The exception is found for the C\u22efC contacts which exhibit positive electrostatic charge for both donor and acceptor atoms signifying the dispersive nature of the contacts.The electrostatic potential property was mapped onto the Hirshfeld surface using the DFT-B3LYP/6-31G of 2.89\u2005\u00c5, Fig.\u00a06e). Further delineation of H\u22efO/O\u22efH, H\u22efC/C\u22efH and H\u22efS/S\u22efH shows that those heterogeneous contacts are more inclined towards -X\u22efH- in DTBA, while the opposite is true for DMF indicating the complementary H-bond accepting and donating nature of DTBA and DMF, respectively. The inclination is more towards -X\u22efH- for (I)The qu\u00adanti\u00adfication of the corresponding close contacts on the Hirshfeld surface through fingerprint plot analysis for overall (I)NCIPLOT 2 and \u03c0(C2\u2013C7)\u2013quasi-\u03c0(O2\u2013C1\u2013O1\u2013H1O\u22efO5\u2013C15\u2013H15) inter\u00adactions as detected in the Hirshfeld surface analysis by calculating the electron density derivatives through wavefunction approach. The visualization of the resulting gradient isosurface supported the existence of the \u03c0\u2013quasi-\u03c0 contacts based on the corresponding large green domain sandwiched between the aromatic and quasi-aromatic rings. The overall density is in the range of \u22120.05 < sign(\u03bb2)\u03c1 < 0.03 a.u. indicating a weak but attractive inter\u00adaction hydrogen bond, leading to the eight-membered homosynthon as well as the seven-membered heterosynthon formed between hy\u00addroxy-O1\u2014H1O\u22efO5(carbon\u00adyl) and DMF-C15\u2014H15\u22efO2(carbon\u00adyl) exhibit the greatest inter\u00adaction energies (Eint) of \u221269.8 and \u221258.9\u2005kJ\u2005mol\u22121, respectively. These are relatively stronger than the other supplementary contacts in (I)viz. electrostatic (Eele), polarization (Epol), dispersion (Edis), exchange-repulsion (Erep) together with the total energy are collated in Table\u00a04The strength of each close contact between all pairwise mol\u00adecules in (I)Crystal Explorer 17, the Eint for the pairs of \u03c0\u22efquasi-\u03c0 inter\u00adactions were modelled in Gaussian16 \u22efquasi-\u03c0(\u22efO4\u2013C14\u2013O3\u2013H3O)2 or \u03c0(C2\u2013C7)\u22efquasi-\u03c0(O2\u2013C1\u2013O1\u2013H1O\u22efO5\u2013C15\u2013H15) inter\u00adactions, i.e. \u221259.3 and \u221259.2\u2005kJ\u2005mol\u22121, respectively.Complementing the calculations with Eele topological framework as shown in Fig.\u00a08a). On the other hand, the dispersion force sustained by the specified \u03c0\u2013\u03c0 inter\u00adactions results in a boat-shape topology, Fig.\u00a08b). The combination of the electrostatic and dispersion forces supersedes the strong inter\u00adaction energy from O\u2014H\u22efO contacts and lead to a refined overall energy framework with razor-blade-like topology, Fig.\u00a08c).The crystal of (I)et al., 2006et al., 2013i.e. occupied unit-cell volume = 220.8\u2005\u00c53 = 25.4% for (I)3 and 47.5% for (II).The crystal structure of DTBA\u00b72DMF (II) is also known, being reported four times In term of contact distribution on the Hirshfeld surface for the corresponding individual DTBA mol\u00adecules and overall (I)Chemical Context, DTBA is usually generated during co-crystallization experiments with 2-mercapto\u00adbenzoic acid (2-MBA), implying oxidation of the latter. In addition to oxidation of 2-MBA, other crystallization outcomes have been observed during recent experiments suggesting chemical reactions are occurring. A less common outcome of crystallization experiments with 2-MBA was the sulfur extrusion product, 2,2\u2032-thiodi\u00adbenzoic acid methyl\u00adene]cyclo\u00adhexane-1,4-di\u00adamine, for n = 2, 3 and 4 was co-crystallized with 2-MBA, a salt of composition [DTBA_2H]\u00b7DMF\u00b7H2O was isolated is 2-(4-ammonio\u00adcyclo\u00adhex\u00adyl)-3-(pyridin-2-yl)imidazopyridin-2-ium di-cation, isolated from the co-crystallization of 2-MBA with the n = 2 isomer of (III) C6H10N(+)H3][4-DTBA_2H]\u00b7DMSO\u00b7H2O was the crystallization product : 3072 \u03bd(C\u2014H), 1680 \u03bd(C=O), 1464 \u03bd(C=C), 1410 \u03b4(C\u2014H), 722 \u03bd(C\u2014S).The DMF monosolvate of DTBA, (I)Uiso(H) set to 1.2Ueq(C). The oxygen-bound H atoms were located from a difference-Fourier map and refined with O\u2014H = 0.84\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989020008257/hb7925sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020008257/hb7925Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020008257/hb7925Isup3.cmlSupporting information file. DOI: 2011285CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Log10 Kw>c, values, of the complementary amino acid pairs are strongly correlated to the central (2nd) purine base of the mRNA codon and the complementary pyrimidine base of the tRNA anticodon. Clustering of amino acids is temperature independent with regard to the direction of translation (3\u2032\u202f\u2192\u202f5\u2032 or 5\u2032\u202f\u2192\u202f3\u2032). The Log10 Kw>c discriminate between artificial Hecht \u03b1- and \u03b2-protein datasets at 25\u00a0\u00b0C and 100\u00a0\u00b0C. Interpretation of this data may be found in the research article entitled \u201cDetermining amino acid scores of the genetic code table: complementarity, structure, function and evolution\u201d .We present the data concerning the clustering of sense and antisense amino acid pairs into polar, nonpolar and neutral groups, as measured using hydrophobicity parameter\u2014logarithmic equilibrium constants (Log Specifications table\u202210 Kw>c) discriminates between artificial \u03b1- and \u03b2-protein datasets at 25\u00a0\u00b0C and 100\u00a0\u00b0C.The data are useful since it is shown that the nucleobase coding of amino acid hydrophobicity, specified by the 2nd codon base, is temperature independent at 25\u00a0\u00b0C and 100\u00a0\u00b0C. The hydrophobicity parameter\u2014logarithmic equilibrium constant nucleobases.\u2022The data presented can be used for theoretical analyses of proteins, experiments with sense and antisense peptide binding, and research of biological systems at different temperature conditions.110 Kw>c10 Kw>c are logarithmic equilibrium constants for the transfer of amino acid side-chains from neutral solution to cyclohexane at 25\u00a0\u00b0C and 100\u00a0\u00b0C The data presented here describe the analysis of temperature dependence of amino acid hydrophobicity parameter\u2014Log10 Kw>c, in a temperature independent manner.10 Kw>c values of the complementary, i.e. sense and antisense, amino acid pairs depend strongly on the central (2nd) purine base of the mRNA codon and the complementary pyrimidine of the tRNA anticodon. All calculated correlations are strong (r\u00a0\u2265\u00a00.85). With respect to the Log10 Kw>c measurements observed, temperatures of 25\u00a0\u00b0C and 100\u00a0\u00b0C do not affect the result specify polar-nonpolar and neutral-neutral clusters for all possible complementary codon pairs irrespective of temperature value and direction of sequence translation.In 10 Kw>c) accurately predict the \u03b1- and \u03b2-artificial protein class at 25\u00a0\u00b0C and 100\u00a0\u00b0C and Phase 2 (secondary) amino acids are clearly separated based on temperature independence of LogPART decision list (25\u00a0\u00b0C)2 AND Log10 Kw>c at 25\u00a0\u00b0C <= 2.64 AND Mean Buried Area > 113.9 \u00c52IF Mean Buried Area > 97.8 \u00c5THEN aa Prebiotic Phase 2 1: L,I,V,S,P,T,A,N,D,E,G)ELSE aa Prebiotic Phase 1 (11/\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014PART decision list (100\u00a0\u00b0C)2 AND Log10 Kw>c at 100\u00a0\u00b0C <= 2.60 AND Mean Buried Area > 113.9 \u00c52IF Mean Buried Area > 97.8 \u00c5THEN aa Prebiotic Phase 2 1: L,I,V,S,P,T,A,N,D,E,G)ELSE aa Prebiotic Phase 1 \u202f=\u202f+1,\u2022polar or hydrophilic amino acid (2nd A codon score)\u202f=\u202f-1,\u2022neutral or intermediate amino acid (2nd C or G codon score)\u202f=\u202f0.The agglomerative hierarchical clustering (HAC) method in High agglomerative coefficients of 0.96 were observed for 25\u00a0\u00b0C and 100\u00a0\u00b0C measurements.2.2k\u202f=\u202f3 clusters, Manhattan metric and standardized variables). The first variable involved the amino acid logarithmic equilibrium constants (Log10 Kw>c), and the second variable was amino acid codon scoring according to Davis Section 2.1.). The values of average silhouette widths for 25\u00a0\u00b0C (0.67) and 100\u00a0\u00b0C (0.65) confirm the validity of the model Two-variable fuzzy partitioning was produced using S-Plus 2000 software (2.310 Kw>c) for transfer of amino acid side-chains from neutral solution to cyclohexane at 25\u00a0\u00b0C and at 100\u00a0\u00b0C. The correlations are presented in https://folk.uio.no/ohammer/past/): x\u202f=\u202ffree energy ligandaa, and y\u202f=\u202f|ligandaa\u202f\u2212\u202freceptoraa| free energy absolute difference (aa\u202f=\u202famino acid).Correlations of complementary pairs of polar-nonpolar residues and neutral-neutral residues in a 3\u2032\u202f\u2192\u202f5\u2032 and 5\u2032\u202f\u2192\u202f3\u2032 translation directions, with respect to the logarithmic equilibrium constants Primary amino acid sequences of 15 artificial Hecht \u03b1- and 17 \u03b2-protein folds were converted into a numerical series by assigning the Log2.610 Kw>c values, and Mean Buried Area parameter https://www.cs.waikato.ac.nz/ml/weka/) The prediction of Phase 1 (primary) and Phase 2 (secondary) amino acids based on temperature independence of Log"} +{"text": "This structure features C\u2014H\u22efO hydrogen bonds (both intra- and inter\u00admolecular) and inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions.The synthesis and crystal structure of a diisopropyl-substituted 13H21NO2S, is reported here along with its crystal structure. This compound crystallizes with two mol\u00adecules in the asymmetric unit. The sulfonamide functional group of this structure features S=O bond lengths ranging from 1.433\u2005(3) to 1.439\u2005(3)\u2005\u00c5, S\u2014C bond lengths of 1.777\u2005(3) and 1.773\u2005(4)\u2005\u00c5, and S\u2014N bond lengths of 1.622\u2005(3) and 1.624\u2005(3)\u2005\u00c5. When viewing the mol\u00adecules down the S\u2014N bond, the isopropyl groups are gauche to the aromatic ring. On each mol\u00adecule, two methyl hydrogen atoms of one isopropyl group are engaged in intra\u00admolecular C\u2014H\u22efO hydrogen bonds with a nearby sulfonamide oxygen atom. Inter\u00admolecular C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions link mol\u00adecules of the title compound in the solid state.The synthesis of the title compound, C We attribute this relatively small torsion angle to the presence of intra\u00admolecular C-H\u22efO inter\u00adactions, which are described in more detail below. The torsion angles (O2\u2014S1\u2014N1\u2014C11 and O2A\u2014S1A\u2014N1A\u2014C11A) between the methine carbon atom of the other isopropyl group and the other sulfonamide oxygen are 46.7\u2005(3) and 46.8\u2005(3)\u00b0, respectively. Both sulfur atoms adopt a slightly distorted tetra\u00adhedral geometry with \u03c44 descriptors for fourfold coordination of 0.94 for both S1 and S1A , we describe them here as potential C\u2014H\u22efO hydrogen bonds. Specifically, O1 inter\u00adacts with C9(H9A) and C10(H10B), while the equivalent atom O1a inter\u00adacts with C9A(H9AC) and C10A(H10F). These inter\u00adactions have D\u22efA distances ranging from 3.039\u2005(5) to 3.157\u2005(5)\u2005\u00c5 and D\u2014H\u22efA angles ranging from 117 to 121\u00b0 x, \u22121\u00a0+\u00a0y, z] with a D\u22efA distance of 3.465\u2005(4)\u2005\u00c5 and a D\u2014H\u22efA angle of 150.8\u00b0 to a stirring mixture of diiso\u00adpropyl\u00adamine , pyridine and 10\u2005mL of degassed di\u00adchloro\u00admethane under a nitro\u00adgen atmosphere. The reaction mixture was stirred at room temperature for 24\u2005h under a nitro\u00adgen atmosphere. After acidification with 5 M HCl and dilution with 15\u2005mL of di\u00adchloro\u00admethane, the organic layer was washed with water and brine. The aqueous layers were back extracted with 10\u2005mL of di\u00adchloro\u00admethane. The combined organic layers were then dried over anhydrous sodium sulfate and evaporated to dryness. The resulting solid was dissolved in hot ethanol and filtered. The filtrate was placed in a freezer for two days and the product was isolated via vacuum filtration to give colorless crystals .The title compound was prepared by the dropwise addition of Uiso(H) = 1.2Ueq(C) for methine groups and aromatic hydrogen atoms, and Uiso(H) = 1.5Ueq(C) for methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020007185/pk2628sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020007185/pk2628Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020007185/pk2628Isup3.cmlSupporting information file. DOI: 2006237CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The 4-iodo\u00adbenzene ring is inclined to the phenol ring by 39.1\u2005(2)\u00b0. The configuration about the C=N bonds is E. The crystal structure features C\u2014H\u22efO hydrogen-bonding inter\u00adactions.In the title Schiff base compound, C 13H9IN2O3, was synthesized by a condensation reaction between 2-hy\u00addroxy-5-nitro\u00adbenzaldehyde and 4-iodo\u00adaniline, and crystallizes in the ortho\u00adrhom\u00adbic space group Pna21. The 4-iodo\u00adbenzene ring is inclined to the phenol ring by a dihedral angle of 39.1\u2005(2)\u00b0. The configuration about the C=N double bond is E. The crystal structure features C\u2014H\u22efO hydrogen-bonding inter\u00adactions. A Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the packing arrangement are O\u22efH/H\u22efO (26.9%) and H\u22efH (22.0%) inter\u00adactions.The title compound, C The imine group displays a C8\u2014C7\u2014N1\u2014C4 torsion angle of 174.5\u2005(6)\u00b0. The 4-iodo\u00adbenzene ring (C1\u2013C6) is inclined by a dihedral angle of 39.1\u2005(2)\u00b0 to the phenol ring (C8\u2013C13), which renders the mol\u00adecule non-planar. The configuration of the C7=N1 bond of this Schiff base is E, and the intra\u00admolecular O1\u2014H1\u22efN1 hydrogen bond forms an S(6) ring motif if Fig.\u00a01. The 4-ni hydrogen bonds between screw-related mol\u00adecules, which form helical chains propagating along the crystallographic screw axis parallel to c x\u00a0+\u00a0y, \u22121\u00a0+\u00a0z)], which makes a zigzag tape motif c Fig.\u00a02. The shoif Fig.\u00a03. There aCrystal Explorer 17.5 or longer than the van der Waals radii sum, respectively rm Fig.\u00a04, white set al., 2016E)-2-{[(4-iodo\u00adphen\u00adyl)imino]\u00admeth\u00adyl}-phenol fragment. Of these 26, the most similar to (I)p-iodo-N-(p-cyano\u00adbenzyl\u00adidene)aniline -5-(di\u00adethyl\u00adamino)-2-[(4-iodo\u00adphenyl\u00adimino)\u00admeth\u00adyl]phenol -4-iodo\u00adbenzene; imino]\u00admeth\u00adyl}-6-meth\u00adoxy\u00adphenol -4-iodo\u00adaniline ring motif.A search of the Cambridge Structural Database and 4-iodo\u00adaniline in ethanol (15\u2005ml). After the mixture had refluxed for about 15\u2005h, the orange product was washed with ether and dried at room temperature . Crystals suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution.Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020008191/pk2635sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020008191/pk2635Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020008191/pk2635Isup3.cmlSupporting information file. DOI: 1922980CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound, containing a tetra-substituted pyrrolidine ring, has an N-bound 4-nitro\u00adphen\u00adyl)ethyl\u00adcarboxyl\u00adate group with an adjacent C-bound 4-meth\u00adoxy\u00adphenyl and then two acet\u00adyloxy subtituents in equatorial and axial positions, respectively. The five-membered ring is twisted about the bond bearing the acet\u00adyloxy subtituents. 23H24N2O9, is a tetra-substituted pyrrolidine derivative with a twisted conformation, with the twist evident in the C\u2014C bond bearing the adjacent acet\u00adyloxy substituents. These are flanked on one side by a C-bound 4-meth\u00adoxy\u00adphen\u00adyl group and on the other by a methyl\u00adene group. The almost sp2-N atom [sum of angles = 357\u00b0] bears a 4-nitro\u00adbenzyl\u00adoxycarbonyl substituent. In the crystal, ring-methyl\u00adene-C\u2014H\u22efO(acet\u00adyloxy-carbon\u00adyl) and methyl\u00adene-C\u2014H\u22efO(carbon\u00adyl) inter\u00adactions lead to supra\u00admolecular layers lying parallel to (The title compound, C The conformation of the five-membered ring is twisted about the C2\u2014C3 bond with the C1\u2014C2\u2014C3\u2014C4 torsion angle being 39.70\u2005(16)\u00b0, consistent with a (+)syn-clinal configuration. The sum of the angles about the N1 atom is 356.7\u00b0, indicating an approximate sp2 centre. The N1-bound group occupies an equatorial position with those at the C1\u2013C3 centres being bis\u00adectional, equatorial and axial, respectively \u00b0.The mol\u00adecular structure of (I)With respect to the least-squares plane through the pyrrolidine ring, the nitro\u00adbenzene and meth\u00adoxy\u00adbenzene rings are splayed, as seen in the dihedral angles of 58.58\u2005(8) and 77.65\u2005(6)\u00b0, respectively; the dihedral angle between the benzene rings is 50.56\u2005(5)\u00b0. There is a twist in the nitro\u00adbenzene ring as seen in the value of the C11\u2014C10\u2014N2\u2014O4 torsion angle of 17.7\u2005(3)\u00b0. By contrast, the meth\u00adoxy group is co-planar with the ring to which it is connected, as shown by the C15\u2014C16\u2014O5\u2014C19 torsion angle of 176.2\u2005(2)\u00b0.b-axis direction, being propagated by 21 symmetry. The other inter\u00adactions falling within the distance criteria of PLATON . A view of the unit-cell contents is shown in Fig.\u00a02b), highlighting the stacking of layers, without directional inter\u00adactions between them.The only directional non-covalent inter\u00adactions of note in the crystal of (I)Supra\u00admolecular features were also evaluated by calculating non-covalent inter\u00adaction plots with the green isosurface between the inter\u00adacting atoms and the distinctive blue feature in the reduced density gradient (RDG) versus sign(\u03bb2)\u03c1(r) plot in the lower view, i.e. indicating the density value is less than 0.0 a.u., suggest these inter\u00adactions are weakly attractive. The same is true for the ring-methyl\u00adene-C4\u2014H\u22efO7(acet\u00adyloxy-carbon\u00adyl) inter\u00adactions that lead to the helical chain, Fig.\u00a03b).The aforementioned weak C\u2014H\u22efO contacts identified in dnorm-surface plots, electrostatic potential and two-dimensional fingerprint plots following literature procedures B as well as the O5 and O7 atoms and O2\u22efO5 short contacts, being \u223c0.02\u2005\u00c5 shorter than their respective sums of the van der Waals radii, Table\u00a02The Hirshfeld surface analysis of (I)B), benzyl (C15 and H9), methyl (C21) and nitro (O4) atoms correspond to long-range intra-layer methyl\u00adene-C6\u2014H6B\u22efC15(benz\u00adyl), benzyl-C9\u2014H9\u22efC21(meth\u00adyl) inter\u00adactions and inter-layer O4\u22efO4 short contacts, Table\u00a02In the views of Fig.\u00a05a), the two-dimensional fingerprint plot for the Hirshfeld surface of (I)de and di diagonal axes, and those delineated into H\u22efH, H\u22efO/O\u22efH, H\u22efC/C\u22efH, O\u22efO and O\u22efC/C\u22efO contacts are illustrated in Fig.\u00a07b)\u2013(f), respectively. The percentage contributions from different inter\u00adatomic contacts are summarized in Table\u00a03de = di \u223c2.4\u2005\u00c5, Fig.\u00a07b), corresponding to the H17\u22efH23B inter-layer contact listed in Table\u00a02de + di \u223c2.5\u2005\u00c5 in Fig.\u00a07c). The H\u22efC/C\u22efH contacts that match the long-range C\u2014H\u22efC inter\u00adactions discussed above are shown as a pairs of forceps-like tips at de + di \u223c2.7\u2005\u00c5 in the fingerprint plot delineated into H\u22efC/C\u22efH contacts, Fig.\u00a07d). Although both O\u22efO and O\u22efC/C\u22efO contacts appear at de + di \u223c3.0\u2005\u00c5 in the respective fingerprint plots, Fig.\u00a07e) and (f), their contributions to the overall Hirshfeld surface are only 2.1 and 1.2%, respectively. The other inter\u00adatomic contacts have a negligible effect on the mol\u00adecular packing as their accumulated contribution is about 2.2%.As illustrated in Fig.\u00a07Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies as per the literature basis set with the B3LYP function. The individual energy components as well as the total inter\u00adaction energies are collated in Table\u00a04Hirshfeld surface analysis as well as two additional C\u2014H\u22efO inter\u00adactions, i.e. methyl\u00adene-C4\u2014H4A\u22efO4(nitro) and methyl-C21\u2014H21C\u22efO4(nitro) with H\u22efO separations of 2.63 and 2.77\u2005\u00c5, respectively.The pairwise inter\u00adaction energies between the mol\u00adecules in the crystal of (I)Edis terms associated with the H\u22efH contacts as well as the long-range C\u2014H\u22efO inter\u00adactions (\u221214.4\u2005kJ\u2005mol\u22121). For the former, the maximum energy is not found for the shortest H17\u22efH23B contact (\u22127.1\u2005kJ\u2005mol\u22121), Table\u00a02b), but rather for a pair of benzene-H\u22efH(meth\u00adyl) inter\u00adactions occurring in close proximity in a hydrogen-rich region but at longer separations (\u221234.2\u2005kJ\u2005mol\u22121). For the inter-layer O4\u22efO4 contact mentioned above, there are almost equal contributions from Eele and Edis, Table\u00a04\u22121. The magnitudes of inter\u00admolecular energies are represented graphically in Fig.\u00a08Edis in the mol\u00adecular packing.The stabilization energies in the inter-layer region are also dominated by the et al., 2016et al., 2004There are relatively few related structures having a similar substitution pattern to the tetra-substituted pyrrolidine ring of (I)S,3S,4R)-3,4-dihy\u00addroxy-2-(4-meth\u00adoxy\u00adphen\u00adyl)pyrrolidine-1-carboxyl\u00adate in CH2Cl2 (15\u2005ml) were added pyridine , acetic anhydride and N,N-dimethyl-4-amino\u00adpyridine . The solution was stirred for 2\u2005h at room temperature, concentrated in a rota-evaporator and the residue dissolved in EtOAc (10\u2005ml). The resulting solution was washed with a HCl 5% solution (3 \u00d7 5\u2005ml) and with saturated solutions of NaHCO3 (2 \u00d7 5\u2005ml) and of NaCl (5\u2005ml). The phases were separated and the organic phase was dried with anhydrous Na2SO4, filtered and concentrated in vacuo.To a solution of 4-nitro\u00adbenzyl (2n-hexane elution gradient (1:3 and 1:2). Yield: 716\u2005mg (98%). Colourless irregular crystals for the X-ray analysis were obtained by the slow evaporation of its n-hexane solution. M.p. 409.5\u2013410.5\u2005K. The 1H and 13C{1H} NMR reflect the presence of two conformational rotamers in solution. 1H NMR : \u03b4 = 7.75 ; 7.65 ; 7.18 ; 6.99 ; 6.76 ; 6.72 ; 6.65 ; 6.37 ; 5.42 ; 5.33 ; 5.00 ; 4.92 ; 4.74 ; 4.44 ; 3.89 ; 3.72 ; 3.29 ; 3.35\u20133.23 ; 1.61\u20131.60 . 1H NMR : \u03b4 = 8.23 ; 8.00 ; 7.53 ; 7.16 ; 6.96 ; 6.88 ; 5.45\u20135.32 ; 5.31\u20135.18 ; 5.01\u20134.87 ; 4.13 ; 4.06 ; 3.85\u20133.67 ; 2.12-2.07 . 13C{1H} NMR : \u03b4 = 169.9; 169.8; 159.4; 159.2; 154.2; 154.1; 147.6; 147.2; 143.6; 143.4; 130.6; 129.4; 128.1; 127.5; 126.8; 126.7; 123.7; 123.4; 114.2; 78.2; 69.2; 68.7; 65.7; 65.5; 64.7; 64.1; 55.3; 55.2; 49.0; 48.4; 20.8; 20.7; 20.6.The residue was purified by flash column chromatography in silica gel, using an EtOAc/Uiso(H) set to 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989020007914/hb7923sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020007914/hb7923Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020007914/hb7923Isup3.cmlSupporting information file. DOI: 2009242CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In a palliative situation like metastatic spinal cord compression (MSCC), overall treatment time of radiotherapy should be as short as possible. This study compared 5\u2009\u00d7\u20095\u00a0Gy in 1\u00a0week to 10\u2009\u00d7\u20093\u00a0Gy in 2\u00a0weeks in a prospective cohort.Forty patients receiving 5\u2009\u00d7\u20095\u00a0Gy in a phase II trial were matched 1:2 to 213 patients receiving 10\u2009\u00d7\u20093\u00a0Gy in two previous prospective studies for tumor type, ambulatory status, time developing motor deficits, interval between tumor diagnosis and MSCC and visceral metastases. These factors were consistent in all three patients (triple) used for each 1:2 matching. Groups were compared for local progression-free survival (LPFS), motor function, ambulatory status, and overall survival (OS).p\u2009=\u20090.36), 6-month OS-rates 43% and 35% (p\u2009=\u20090.74). Improvement of motor function was achieved in 59% and 34% of patients (p\u2009=\u20090.028); overall response rates (improvement or no further progression of motor deficits) were 94% and 89% (p\u2009=\u20090.71). Post-treatment ambulatory rates were 81% after 5\u2009\u00d7\u20095\u00a0Gy and 85% after 10\u2009\u00d7\u20093\u00a0Gy (p\u2009=\u20090.61). Of non-ambulatory patients, 50% (6/12) and 46% (11/24) regained the ability to walk (p\u2009=\u20091.00).After matching, 32 triples remained for analyses . Six-month LPFS-rates were 94% after 5\u2009\u00d7\u20095\u00a0Gy and 87% after 10\u2009\u00d7\u20093\u00a0Gy (5\u2009\u00d7\u20095\u00a0Gy in 1\u00a0week appeared similarly effective as 10\u2009\u00d7\u20093\u00a0Gy in 2\u00a0weeks. These results may not be applicable to long-term survivors and should be confirmed in a randomized trial directly comparing 5\u2009\u00d7\u20095\u00a0Gy and 10\u2009\u00d7\u20093\u00a0Gy.Trial registration clinicaltrials.gov NCT03070431. Registered 27 February 2017. Metastatic spinal cord compression (MSCC) occurs in 5\u201310% of patients with malignant diseases . Many ofPatients in the phase II trial received precision radiotherapy with 5\u2009\u00d7\u20095\u00a0Gy in 1\u00a0week between 02/2017 and 03/2018 . ImproveSince the EQD2 of 5\u2009\u00d7\u20095\u00a0Gy (31.3\u00a0Gy) for tumor cell kill is similar to 10\u2009\u00d7\u20093\u00a0Gy (32.5\u00a0Gy), it is assumed that both regimens are similarly effective . PatientThe 40 patients of the PRE-MODE trial were matched 1:2 to the 213 patients receiving 10\u2009\u00d7\u20093\u00a0Gy in a previous trial , 10. MatComparisons for LPFS and OS were performed using Kaplan\u2013Meier method and log-rank test. For the comparison regarding age, the Mann-Whiney U test was used. The comparisons for patient characteristics, improvement of motor function, overall response and ambulatory status were performed with the Fisher\u2019s exact test.z-score was 0.933, and distribution was considered approximately normal.After 1:2 matching, 32 triples remained for analyses, corresponding to a total of 96 patients. The distribution of the patient characteristics was not significantly different (Table p\u2009=\u20090.36), and 6-month OS-rates 43% and 35% . Improvement of motor function was achieved in 59% (19/32) and 34% (22/64) of patients (p\u2009=\u20090.028). Overall response rates were 94% (30/32) and 89% (57/64) (p\u2009=\u20090.71). Deterioration of motor deficits occurred in 6% (2/32) and 11% (7/64) of patients (p\u2009=\u20090.71). Post-treatment ambulatory rates were 81% after 5\u2009\u00d7\u20095\u00a0Gy and 85% after 10\u2009\u00d7\u20093\u00a0Gy (p\u2009=\u20090.61). Of non-ambulatory patients, 50% (6/12) after 5\u2009\u00d7\u20095\u00a0Gy and 46% (11/24) after 10\u2009\u00d7\u20093\u00a0Gy, respectively, regained walking ability (p\u2009=\u20091.00). Of ambulatory patients, 100% (20/20) and 93% (37/40), respectively, remained ambulatory (p\u2009=\u20090.54).Six-month LPFS-rates were 94% after 5\u2009\u00d7\u20095\u00a0Gy and 87% after 10\u2009\u00d7\u20093\u00a0Gy when compared to a historical control group receiving 5\u2009\u00d7\u20094\u00a0Gy [Important endpoints of radiotherapy for MSCC include functional outcome and local control \u20133. In se5\u2009\u00d7\u20094\u00a0Gy . EQD2 fo5\u2009\u00d7\u20094\u00a0Gy . 5\u2009\u00d7\u20095\u00a0GThis study compared 5\u2009\u00d7\u20095\u00a0Gy to 10\u2009\u00d7\u20093\u00a0Gy using data of three prospective studies. Forty patients receiving 5\u2009\u00d7\u20095\u00a0Gy were matched 1:2 to 213 patients receiving 10\u2009\u00d7\u20093\u00a0Gy. Matching criteria were selected according to previous studies , 12\u201314. 5\u2009\u00d7\u20095\u00a0Gy resulted in a significantly higher rate of improvement of motor function than 10\u2009\u00d7\u20093\u00a0Gy. For the other investigated endpoints including the main objective LPFS, no significant differences were found. Thus, 5\u2009\u00d7\u20095\u00a0Gy appeared similarly effective as 10\u2009\u00d7\u20093\u00a0Gy. When interpreting these results, the limitations of the study must be considered including the non-randomized design. To minimize the risk of hidden selection biases, we used only data from prospective studies. Further limitations include the facts that follow-up MRI was not performed at pre-defined time points and that for grading of MSCC radiological criteria were not considered \u201310, 15. 5\u2009\u00d7\u20095\u00a0Gy in 1\u00a0week appeared similarly effective as 10\u2009\u00d7\u20093\u00a0Gy in 2\u00a0weeks for LPFS, functional outcome and OS. These results may not be applicable to long-term survivors and should be confirmed in a randomized trial that directly compares 5\u2009\u00d7\u20095\u00a0Gy and 10\u2009\u00d7\u20093\u00a0Gy."} +{"text": "The aim of the study was to explore genotype distribution thalassemia and G6PD deficiency in Meizhou city, China.G6PD mutations by gene chip analysis. Genotypes and allele frequencies were analyzed.A total of 16\u00a0158 individuals were involved in thalassemia genetic testing. A total of 605 subjects were screened for common Chinese SEA (65.12%), \u2010\u03b13.7 (19.05%), and \u2010\u03b14.2 (8.05%) deletion were the main mutations of \u03b1\u2010thalassemia, while IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T) (40.39%), CD41\u201042(\u2010TCTT) (32.72%), \u201028(A\u00a0\u2192\u00a0G) (10.11%), and CD17(A\u00a0\u2192\u00a0T) (9.32%) mutations were the principal mutations of \u03b2\u2010thalassemia in Meizhou. There were significant differences in allele frequencies in some counties. Genetic testing for G6PD deficiency, six mutation sites, and one polymorphism were detected in our study. A total of 198 alleles with the mutation were detected among 805 alleles (24.6%). G6PD Canton (c.1376 G\u00a0\u2192\u00a0T) (45.96%), G6PD Kaiping (c.1388 G\u00a0\u2192\u00a0A) (39.39%), and G6PD Gaohe (c.95 A\u00a0\u2192\u00a0G) (9.09%) account for 94.44% mutations, followed by G6PD Chinese\u20105 (c.1024 C\u00a0\u2192\u00a0T) (4.04%), G6PD Viangchan (c.871G\u00a0\u2192\u00a0A) (1.01%), and G6PD Maewo (c.1360 C\u00a0\u2192\u00a0T) (0.51%). There were some differences of the distribution of G6PD mutations among eight counties in Meizhou.A total of 5463 cases carried thalassemia mutations were identified, including 3585 cases, 1701 cases, and 177 cases with \u03b1\u2010, \u03b2\u2010, and \u03b1\u00a0+\u00a0\u03b2\u2010thalassemia mutations, respectively. \u2010\u2010SEA, \u2010\u03b13.7, and \u2010\u03b14.2 deletion were the main mutations of \u03b1\u2010thalassemia, while IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T), CD41\u201042(\u2010TCTT), \u201028(A\u00a0\u2192\u00a0G), and CD17(A\u00a0\u2192\u00a0T) mutations were the principal mutations of \u03b2\u2010thalassemia in Meizhou. G6PD c.1376 G\u00a0\u2192\u00a0T, c.1388 G\u00a0\u2192\u00a0A, and c.95 A\u00a0\u2192\u00a0G were the main mutations of G6PD deficiency. There were some differences of the distribution of thalassemia and G6PD mutations among eight counties in Meizhou.The \u2010\u2010 As one of the commonest monogenic disorder in the world, thalassemia assumes diversity in clinical phenotypes varying from almost asymptomatic to lethal hemolytic anemia.G6PD gene is located on chromosome Xq28 which consists of 13 exons and 12 introns, encoding 515 amino acids. The deficiency is widely distributed and occurs in about 400\u00a0million people worldwide.Hereditary G6PD deficiency is one of the most common genetic enzyme deficiency diseases in the world. G6PD deficiency is an X\u2010linked incomplete dominant inherited\u00a0disease. The Meizhou is a city located in the northeast of Guangdong Province, and most of the residents living in this area are Hakka peoples. Hakka is an intriguing Han Chinese population that mainly inhabit in southern China who migrated to south originally from northern China.Population screening and genetic counseling are important to prevent the birth of children with thalassemia major. Using genetic analysis for prenatal diagnosis can diagnose thalassemia major fetuses in early pregnancy and terminate pregnancy in time, so as to avoid the birth of thalassemia major patients, which is an effective method to prevent this disease at present. Precise mutation frequencies studies in different populations will help healthcare programs to control thalassemia.22.1A total of 16\u00a0158 individuals who visited Meizhou People's Hospital from January 2015 to May 2018 were involved in thalassemia genetic testing in this study. From February 2016 to May 2018, 605 subjects were screened for common Chinese G6PD mutations by gene chip analysis. The subjects included patients who went to cardiovascular disease center, prenatal diagnosis center, reproductive medicine center, physical examination center, pediatrics, gynecology, and other professional departments of our hospital, excluding patients with blood diseases. Figure 2.22.2.12 (<2.5%) were considered probable \u03b1\u2010thalassemia carriers, whereas with high HbA2 (>3.5%) were deemed possible \u03b2\u2010thalassemia carriers.Samples were obtained via venipuncture of an antecubital vein, and then, 2\u00a0mL of peripheral blood was collected in EDTA anticoagulant tube. Sysmex XE\u20102100 blood analyzer was used to determine erythrocyte correlative indices following the standard operating procedures. Hemoglobin electrophoresis analysis was performed by Sebia capillary electrophoresis system compiling with standard operating procedures. Subjects detected to low mean corpuscular volume (MCV) values (<82\u00a0fL) and (or) lower mean corpuscular hemoglobin (MCH) values (<27\u00a0pg) were thought as suspicious thalassemia carriers. Subjects with low HbA2.2.2SEA, \u2010\u03b13.7, and \u2010\u03b14.2) and non\u2010deletional mutations (Hb Constant Spring (\u03b1CS\u03b1) , Hb Quong Sze (\u03b1QS\u03b1) , and Hb Westmead (\u03b1WS\u03b1) ). Sixteen common non\u2010deletional mutations in \u03b2\u2010globin gene were detected by PCR and flow\u2010through hybridization technology : CD41\u201042(\u2010TCTT), CD43(G\u00a0\u2192\u00a0T), IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T), CD17(A\u00a0\u2192\u00a0T), CD14\u201015(+G), \u201028(A\u00a0\u2192\u00a0G), \u221229(A\u00a0\u2192\u00a0G), CD71\u201072(+A), CD26(G\u00a0\u2192\u00a0A), IVS\u2010I\u20101(G\u00a0\u2192\u00a0T), IVS\u2010I\u20101(G\u00a0\u2192\u00a0A), CD27\u201028(+C), IVS\u2010I\u20105(G\u00a0\u2192\u00a0C), Cap\u00a0+\u00a040\u201043(\u2010AAAC), initiation codon (T\u00a0\u2192\u00a0G), and CD31(\u2010C).Peripheral blood was collected with EDTA anticoagulant tube, in which genomic DNA was extracted from leukocytes. DNA concentration was quantified using NanoDrop 2000\u2122 Spectrophotometer (Thermo Fisher Scientific). Gap\u2010polymerase chain reaction (gap\u2010PCR) and flow\u2010through hybridization technology were used to detect \u03b1\u2010thalassemia mutations, both deletional mutations (\u2010\u20102.3G6PD Gene Typing Detection Kit (gene chip assay) (Sinochips Bioscience Co.). We detected the six mutation and one polymorphism sites in G6PD gene most commonly seen in the Chinese population by gene chip assay, including G6PD Gaohe , G6PD Viangchan , G6PD Chinese\u20105 , G6PD Maewo , G6PD Canton , G6PD Kaiping , and one polymorphism .Genomic DNA was extracted from EDTA anticoagulant blood of subjects using the QIAamp DNA Blood Mini Kit (Qiagen). Amplification was performed using the 2.4P\u00a0<\u00a0.05 was considered to statistical difference.This study used SPSS statistical software version 20.0 to analyze data, and the results would be displayed with corresponding proportion. Descriptive analysis and Pearson chi\u2010square test were used to compare the frequencies of genotype and allele among different counties in Meizhou region. 3A total of 16\u00a0158 blood samples were obtained and analyzed from eight counties of Meizhou area. A total of 8701 cases with microcytosis (MCV\u00a0<\u00a082\u00a0fL and/or MCH\u00a0<\u00a027\u00a0pg) were found, and corresponding percentages of microcytosis in Xingning, Wuhua, Meixian, Meijiang, Fengshun, Jiaoling, Pingyuan, and Dabu were 56.96% (1792/3146), 56.87% (1750/3077), 51.29% (1407/2743), 55.14% (1025/1859), 48.76% (1023/2098), 48.33% (652/1349), 55.54% (536/966), and 55.98% (515/920), respectively. The corresponding abnormal\u00a0ratio\u00a0of\u00a0hemoglobin in Xingning, Wuhua, Meixian, Meijiang, Fengshun, Jiaoling, Pingyuan, and Dabu were 52.64% (1656/3146), 52.00% (1600/3077), 46.34% (1271/2743), 47.98% (892/1859), 47.47% (996/2098), 44.48% (600/1349), 48.96% (473/966), and 50.98% (469/920), respectively.P\u00a0=\u00a0.043) was lower than other counties in Meizhou area. However, the relative proportions of \u03b1\u2010 and \u03b2\u2010thalassemia did not differ greatly among eight counties. The prevalence of \u03b1\u2010thalassemia in Meijiang was lower than other counties in Meizhou area. The prevalence of Hb H disease in Wuhua was higher than other counties in Meizhou area, while in Jiaoling was lower than other counties.The result of the incidence rate of \u03b1\u2010 and \u03b2\u2010thalassemia of eight counties in Meizhou area is shown in Table P\u00a0=\u00a0.032) and \u2010\u03b13.7 deletion , and higher frequencies in \u03b1WS\u03b1 allele of Xingning , in \u2010\u03b14.2 deletion of Jiaoling , in \u03b1CS\u03b1 allele of Dabu , and in IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T) allele of Pingyuan were showed compared with other counties separately. IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T) mutation was found dominance in most of these counties, while Meixian and Fengshun had a higher percentage in CD41\u201042(\u2010TCTT) allele. The \u03b2\u2010thalassemia mutations in the order of allele frequency between eight regions of Meizhou, the mutations were IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T)\u00a0>\u00a0CD41\u201042(\u2010TCTT)\u00a0>\u00a0\u201028(A\u00a0\u2192\u00a0G)\u00a0>\u00a0CD17(A\u00a0\u2192\u00a0T) in Xingning, Meijiang, Jiaoling and Pingyuan (similar to Jiangxi), IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T)\u00a0>\u00a0CD41\u201042(\u2010TCTT)\u00a0>\u00a0CD17(A\u00a0\u2192\u00a0T)\u00a0>\u00a0\u201028(A\u00a0\u2192\u00a0G) in Wuhua and Dabu (similar to Fujian), CD41\u201042(\u2010TCTT)\u00a0>\u00a0IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T)\u00a0>\u00a0CD17(A\u00a0\u2192\u00a0T)\u00a0>\u00a0\u201028(A\u00a0\u2192\u00a0G) in Meixian and Fengshun.Allele frequencies of \u03b1\u2010 and \u03b2\u2010thalassemia of eight counties in Meizhou area are exhibited in Table 4.2/\u03b1\u03b1, \u03b2N/\u03b2N of Jiaoling , in \u2010\u2010SEA/\u2010\u03b13.7, \u03b2N/\u03b2N and \u03b1\u03b1/\u03b1\u03b1, \u03b2IVS\u2010II\u2010654/\u03b2N of Pingyuan, in \u03b1CS\u03b1/\u03b1\u03b1, \u03b2N/\u03b2N of Meixian , and in \u2010\u2010SEA/\u03b1CS\u03b1, \u03b2N/\u03b2N of Dabu compared with other counties. There are lower frequencies in \u03b1CS\u03b1/\u03b1\u03b1, \u03b2N/\u03b2N of Xingning and in \u03b1\u03b1/\u03b1\u03b1, \u03b2\u201028/\u03b2N of Dabu compared with other counties.The result of genotypes of different counties in Meizhou area is shown in Table G6PD mutations and 19 genotypes were detected, including six kinds of heterozygotes, six kinds of hemizygotes, and seven kinds of homozygotes. The result of allele frequencies of G6PD in different counties of Meizhou area is shown in Table G6PD mutations including G6PD Canton (c.1376 G\u00a0\u2192\u00a0T)(45.96%), G6PD Kaiping (c.1388 G\u00a0\u2192\u00a0A)(39.39%), and G6PD Gaohe (c.95 A\u00a0\u2192\u00a0G)(9.09%) account for 94.44% of mutations, followed by G6PD Chinese\u20105 (c.1024 C\u00a0\u2192\u00a0T)(4.04%), G6PD Viangchan (c.871G\u00a0\u2192\u00a0A)(1.01%), and G6PD Maewo (c.1360 C\u00a0\u2192\u00a0T)(0.51%). There were higher frequency in G6PD c.1376 G\u00a0\u2192\u00a0T allele of Wuhua and lower frequency in G6PD c.1376 G\u00a0\u2192\u00a0T allele of Meijiang compared with other counties. The mutations were c.1376 G\u00a0\u2192\u00a0T\u00a0>\u00a0c.1388 G\u00a0\u2192\u00a0A\u00a0>\u00a0c.95 A\u00a0\u2192\u00a0G in Xingning and Dabu, c.1376 G\u00a0\u2192\u00a0T\u00a0>\u00a0c.1388 G\u00a0\u2192\u00a0A\u00a0>\u00a0c.95 A\u00a0\u2192\u00a0G in Wuhua and Meixian, c.1388 G\u00a0\u2192\u00a0A\u00a0>\u00a0c.1376 G\u00a0\u2192\u00a0T\u00a0>\u00a0c.1024 C\u00a0\u2192\u00a0T in Meijiang, c.1388 G\u00a0\u2192\u00a0A\u00a0>\u00a0c.1376 G\u00a0\u2192\u00a0T\u00a0>\u00a0c.95 A\u00a0\u2192\u00a0G in Fengshun, c.1388 G\u00a0\u2192\u00a0A\u00a0>\u00a0c.1376 G\u00a0\u2192\u00a0T\u00a0>\u00a0c.1024 C\u00a0\u2192\u00a0T in Jiaoling, and c.1376 G\u00a0\u2192\u00a0T\u00a0>\u00a0c.1388 G\u00a0\u2192\u00a0A\u00a0>\u00a0c.1360 C\u00a0\u2192\u00a0T in Pingyuan. In addition, we detected 55 patients (9.09%) with the polymorphism (c.1311 C\u00a0\u2192\u00a0T).For G6PD deficiency, there were six Among these cases, 130 cases were simultaneously performed on genetic testing for thalassemia and G6PD deficiency, among them, 70 cases with G6PD deficiency mutations, 27 cases carried thalassemia mutations were identified, and 13 cases with both thalassemia and G6PD deficiency mutations\u00a0and\u00a046 cases did not carry thalassemia and G6PD deficiency mutations. Due to the small number of cases, we cannot analyze the relationship of thalassemia and G6PD deficiency. Of course, further studies are required to reveal the relationship, which is one of our next major research goals.4Thalassemia is an inherited autosomal recessive disease that is mainly prevalent in Guangdong, Guangxi, and Hainan provinces. Thalassemia major places a heavy financial burden on some families.In this study, the total incidence of thalassemia in Meizhou region was 33.80%, of \u03b1\u2010thalassemia was 22.17%, of \u03b2\u2010thalassemia was 10.53%, and of \u03b1\u2010compound \u03b2\u2010thalassemia was 1.10%. Compared with previous studies, our research presented relatively higher prevalence of thalassemia because our subjects were hospitalized patients.SEA/\u03b1\u03b1 and 39.4% of \u03b2\u2010thalassemia with the main genotype of CD41\u201042(\u2010TCTT) and IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T) (5.8%) ranked the fifth position.4.2, \u2010\u03b13.7 deletion, 3.83% of \u03b2\u2010thalassemia with lead genotype of CD41\u201042(\u2010TCTT) mutation, while the Han people in Hainan Province presented 12.16% of \u03b1\u2010thalassemia with chief genotype of \u2010\u2010SEA, \u2010\u03b13.7 deletion, 6.11% of \u03b2\u2010thalassemia with the main mutation is CD41\u201042(\u2010TCTT).SEA and \u2010\u03b13.7 deletion were the main genotypes with frequencies 46.9% and 36.5%, respectively, while the carrier rate of \u03b2\u2010thalassemia was 2.54% in which CD41\u201042(\u2010TCTT) (36.4%) and IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T) (24.8%) were the principal genotypes.SEA deletion (74.85% of mutations), 1.99% of \u03b2\u2010thalassemia in which CD41\u201042(\u2010TCTT) and IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T) played a dominant role with frequencies 41.25% and 27.50%, respectively.SEA deletion was 3.44%, 3.97% of \u03b2\u2010thalassemia with chief genotype of CD41\u201042(\u2010TCTT).SEA/\u03b1\u03b1 was the most common \u03b1\u2010thalassemia genotype, accounted for 62.93% of \u03b1\u2010thalassemia. \u03b1WS\u03b1 accounted for 0.37% (120/32316) of \u03b1\u2010thalassemia and was relatively lower than the proportion of in Guangdong Province reported in the previous study. The main genotype of \u03b2\u2010thalassemia in Meizhou is IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T), whereas the counterpart in Guangdong is CD41\u201042(\u2010TCTT); however, CD41\u201042(\u2010TCTT) showed higher proportion in Fengshun and Meixian.The statistics and analysis of molecular characteristics and morbidity of thalassemia in different regions have increasingly enriched in many studies. For example, frequency of carriers for \u03b1\u2010thalassemia is 15% and \u03b2\u2010thalassemia carriers comprise 4.8% in Guangxi Province, while 46.7% of \u03b1\u2010thalassemia with the main genotype of \u2010\u2010SEA deletion is potentially at risk of begetting infants with HbH disease or the Hb Bart's hydrops fetalis syndrome, depending upon the \u03b1\u2010globin genotype of his/her partner, which would exhibit range from hypochromic anemia to hydrops fetalis. According to previous researches, most of regions indicated a similar feature with higher genotype of \u2010\u2010SEA deletion. Our results are consistent with those reported in most other regions. Nonetheless, the distributions and proportions in different areas regarding \u03b2\u2010thalassemia genotype seem to be subtly difference. Our research may not fully represent the epidemiology of thalassemia in this region related to the randomization and sample capacity of study subjects possibly as for most of our subjects were hospitalized patients.A person who is a carrier of \u2010\u2010G6PD mutations in Hakka population of Guangdong Province studied by Yu et alG6PD gene mutations in Chinese population, which generally accounts for more than 50%. In this study, c.1376 G\u00a0\u2192\u00a0T, c.1388 G\u00a0\u2192\u00a0A, and c.95 A\u00a0\u2192\u00a0G mutations accounted for 94.44% of the total mutation types, and the mutation distribution was consistent with the results in other places.G6PD deficiency gene carrying incidence rate was 5.41% in Guangdong Province.G6PD gene mutations, and next study may reveal the correlation between thalassemia and G6PD deficiency.It is reported that the G6PD enzyme activity in patients with G6PD deficiency compound thalassemia may be higher than that of G6PD deficiency patients, due to an increase in the number of newborn erythrocytes and increase in the activity of G6PD in the chronic hemolysis of the G6PD deficiency compound thalassemia patients.The genotype distribution and frequencies of thalassemia and G6PD deficiency in Meizhou area have regional characteristics, and there are also significant differences in different counties. Local governments can formulate corresponding measures and detection projects to prevent and control thalassemia major and G6PD deficiency according to the genotype distribution and frequencies, effectively saving costs and enhancing social benefits.5SEA, \u2010\u03b13.7, and \u2010\u03b14.2 deletion were the main mutations of \u03b1\u2010thalassemia, while IVS\u2010II\u2010654(C\u00a0\u2192\u00a0T), CD41\u201042(\u2010TCTT), \u201028(A\u00a0\u2192\u00a0G) and CD17(A\u00a0\u2192\u00a0T) mutations were the principal mutations of \u03b2\u2010thalassemia in Meizhou. Genetic testing for G6PD deficiency, c.1376 G\u00a0\u2192\u00a0T, c.1388 G\u00a0\u2192\u00a0A, and c.95 A\u00a0\u2192\u00a0G were the main mutations of G6PD deficiency in this region. In addition, there were some differences of the distribution of thalassemia and G6PD mutations among eight counties in Meizhou.In conclusion, \u2010\u2010"} +{"text": "N atom generating an S(6) ring motif. The 3-chloro\u00adbenzene ring is inclined to the phenol ring by 9.38\u2005(11)\u00b0. The configuration about the C=N bond is E.In the title Schiff base compound, the hy\u00addroxy group forms a intra\u00admolecular hydrogen bond to the imine 15H14ClNO, was synthesized by condensation reaction of 2-hy\u00addroxy-5-methyl\u00adbenzaldehyde and 3-chloro-4-methyl\u00adaniline, and crystallizes in the monoclinic space group P21/c. The 3-chloro\u00adbenzene ring is inclined to the phenol ring by 9.38\u2005(11)\u00b0. The configuration about the C=N bond is E and an intra\u00admolecular O\u2014H\u22efN hydrogen bond forms an S(6) ring motif. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the packing arrangement are from H\u22efH (43.8%) and C\u22efH/H\u22efC (26.7%) inter\u00adactions. The density functional theory (DFT) optimized structure at the B3LYP/ 6\u2013311\u2005G level is compared with the experimentally determined mol\u00adecular structure and the HOMO\u2013LUMO energy gap is provided.The title compound, C RCH=N\u2013R\u2032) and are prepared by condensation reactions between amines and active carbonyl compounds. Schiff bases are employed as catalyst carriers . The imine group, which displays a C9\u2014C8\u2014 N1\u2014C5 torsion angle of \u2212177.49\u2005(18)\u00b0, contributes to the general non-planarity of the mol\u00adecule. The chloro\u00adbenzene ring (C2\u2013C7) is inclined by 9.38\u2005(11)\u00b0 to the phenol ring (C9\u2013C14). The configuration of the C7=N1 bond of this Schiff base is E, and the intra\u00admolecular O1\u2014H1\u22efN1 hydrogen bond forms an S(6) ring motif A\u22efN1 [H1A\u22efN1 = 2.86\u2005\u00c5] inter\u00adactions, forming sheets propagating along the a-axis direction . Weak C\u2014H\u22ef\u03c0 inter\u00adactions [C1\u2014H1C\u22efCg1 = 2.92\u2005\u00c5] are observed (Table\u00a01b). Notably, weak \u03c0\u2013\u03c0 stacking inter\u00adactions between chloro\u00adbenzene rings along the a axis lead to the formation of a three-dimensional network.In the crystal packing of (I)on Fig.\u00a02a. Weak d Table\u00a01b. NotabCrystal Explorer 17.5 and shape index (e) surface mappings are shown in Fig.\u00a03dnorm surface represent O1\u22efCl1 inter\u00adactions and C1\u2014H1C\u22efCg1 inter\u00adactions . Some additional inter\u00adactions indicated by light-red spots are corresponding to contacts around phenolic and chloro\u00adbenzene rings . The red and blue triangles are absent on the shape-index surface, which indicates there are no strong \u03c0\u2013\u03c0 stacking inter\u00adactions in the crystal structure.The inter\u00admolecular inter\u00adactions were investigated qu\u00adanti\u00adtatively and visualized with ns Fig.\u00a03b and C1ns Fig.\u00a03c. Some gs Fig.\u00a03d. The ra\u2013f) indicates that the H\u22efH (43.8%) inter\u00adactions are the major factor in the crystal packing with C\u22efH/H\u22efC (26.7%) inter\u00adactions making the next highest contribution. The percentage contributions of other weak inter\u00adactions are: Cl\u22efH/H\u22efCl (12.4%), O\u22efH/H\u22efO (6.6%) and N\u22efH/H\u22efN (3.8%).Analysis of the two-dimensional fingerprint plots Fig.\u00a04a\u2013f indivia density functional theory (DFT) using standard B3LYP functional and 6\u2013311\u2005G basis-set calculations , hardness (\u03b7), electrophilicity (\u03c9), softness (\u03c3) and fraction of electron transferred (\u0394N). These data are recorded in Table\u00a03\u03c3 is for the evaluation of both the reactivity and stability. The electron transition from the HOMO to the LUMO energy level is shown in Fig.\u00a05E = ELUMO\u00a0\u2212\u00a0EHOMO] of the mol\u00adecule is 4.0023\u2005eV, the frontier mol\u00adecular orbital energies EHOMO and ELUMO being \u22125.9865\u2005eV and \u22121.9842\u2005eV, respectively. The dipole moment of (I)The optimized structure in the gas phase of compound (I)et al., 2016E)-4-meth\u00adoxy-2-{[(4-methyl\u00adphen\u00adyl)imino]\u00admeth\u00adyl}phenol -(5-chloro-2-methyl\u00adphen\u00adyl)imino\u00admeth\u00adyl]-4-methyl\u00adphenol -[(3-chloro-4-meth\u00adyl\u00adphen\u00adyl)imino]\u00admeth\u00adyl}-4-(tri\u00adfluoro\u00admeth\u00adoxy)phenol ring. In 2-{(E)-[(3-iodo-4-methyl\u00adphen\u00adyl)imino]\u00admeth\u00adyl}-4-(tri\u00adfluoro\u00admeth\u00adoxy)phenol -2,4,6-tri\u00admethyl\u00adaniline in ethanol (15\u2005ml) and 3-chloro-4-methyl\u00adaniline in ethanol (15\u2005ml). The reaction mixture was stirred for 5\u2005h under reflux. Single crystals of the title compound suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution .H atom was located in a difference-Fourier map and its positional parameters were refined freely with Uiso(H) = 1.5Ueq(O). Other H atoms were fixed geometrically and treated as riding with C\u2014H = 0.96\u2005\u00c5 (meth\u00adyl) or 0.93\u2005\u00c5 (aromatic), and Uiso(H) = 1.2Ueq(C) for aromatic H atoms or Uiso(H) = 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989020009421/vm2236sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020009421/vm2236Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020009421/vm2236Isup3.cmlSupporting information file. DOI: 2015356CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Ohy\u00addroxy\u2014H\u22efOwater, Ohy\u00addroxy\u2014H\u22efOhy\u00addroxy, Owater\u2014H\u22efOhy\u00addroxy, and Owater\u2014H\u22efNpyridine, result in the formation of a ribbon structure running along .In the title pyridine derivative, C In this paper, we report the crystal structure of 2,6-bis\u00ad(3-methyl\u00adbutyn-3-ol)pyridine monohydrate, 1\u00b7H2O.Pyridine derivatives with propargyl alcohol groups as substituents in the 2,6-positions are inter\u00adesting compounds that have been used as synthons of many reactive compounds and C\u2261C\u2013C(OH) (C6\u2261C7\u2014C8 and C11\u2261C12\u2014C13) bond angles are 176.0\u2005(2), 176.4\u2005(2), 174.6\u2005(2) and 178.5\u2005(2)\u00b0, respectively. C6\u2261C7\u2014C8 is slightly distorted from a linear structure compared to the other bonds. The two OH groups are oriented in directions opposite to each other with respect to the plane of the pyridine ring, and the pyridine ring makes dihedral angles of 50.50\u2005(17) and 57.58\u2005(15)\u00b0, respectively, with the C7/C8/O1 and C12/C13/O2 planes.The mol\u00adecular structure of the title compound is depicted in Fig.\u00a011\u00b7H2O along the c axis. The water mol\u00adecules present as the crystallization solvent form inter\u00admolecular O\u2014H\u22efO and O\u2014H\u22efN inter\u00adactions with the hydroxyl groups and the N atoms of the pyridine unit of mol\u00adecule 1 \u2212x\u00a0+\u00a0y\u00a0+\u00a01, z\u00a0+\u00a01 have been reported 2 as a catalyst in a THF (50\u2005mL)\u2013NEt3 (150\u2005mL) solvent for 19\u2005h at room temperature. The resulting dark-brown solution was quenched with an aqueous NH4Cl solution and the obtained solid was elimin\u00adated by celite filtration. The solution was extracted by AcOEt, and the organic phase was dried over MgSO4. After filtering off the desiccant, the filtrate was concentrated and subjected to silica-gel chromatography (eluent: AcOEt:hexane 3:2). Single crystals suitable for X-ray diffraction studies were obtained from an ethyl acetate solution via slow evaporation in air.2,6-Bis(3-methyl\u00adbutyn-3-ol)pyridine was prepared by using a modified Potts method (Potts Uiso(H) = 1.2Ueq (aromatic-C) or 1.5Ueq (methyl-C).Crystal data, data collection and refinement details are summarized in Table\u00a0210.1107/S2056989020013304/is5553sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020013304/is5553Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020013304/is5553Isup3.cmlSupporting information file. DOI: 2035321, 2035321CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The bridged bimetallic compounds adopt \u039b-shaped configurations, with the octa\u00adhedrally coordinated copper(II) center linked to the fluorinated early transition metal via a fluoride linkage. The extended structures of these \u039b-shaped compounds are organized through both intra- and inter\u00admolecular hydrogen bonds and inter\u00admolecular \u03c0\u2013\u03c0 stacking. The salt compound [Cu(phen)(H2O)2F]2[HfF6]\u00b7H2O displays an isolated square-pyramidal Cu(phen)(H2O)2F+ complex linked to other cationic complexes and isolated HfF62\u2212 anions through inter\u00admolecular hydrogen-bonding inter\u00adactions.The crystal structures of three bridged bimetallic mol\u00adecular compounds, namely, tri\u00adaqua-2\u03ba Although these compounds crystallize with inversion symmetry, the novel mol\u00adecular building units are potential targets of future studies aimed to perturb their packing arrangement to form NCS structures. The salt compound [Cu(phen)(H2O)2F]2[HfF6]\u00b7H2O provides a point of comparison as an unbridged analogue of [Cu(phen)(H2O)3(HfF6)]\u00b7H2O.Lambda (\u039b)-shaped mol\u00adecules have been demonstrated as efficient building blocks in the synthesis of non-centrosymmetric (NCS) materials 2O)3(TiF6)]\u00b7H2O and crystallizes in the ortho\u00adrhom\u00adbic space group Pbca to 1.9035\u2005(13)\u2005\u00c5. The \u039b-shape, indicated by the Cu1\u2014F1\u2014Ti1 bond angle of 134.93\u2005(6)\u00b0, is enforced by the two intra\u00admolecular O2\u2014H2B\u22efF5 and O3\u2014H3B\u22efF6 hydrogen bonds (Table\u00a01Compound (I)ca Fig.\u00a01. The strs Table\u00a01.2O)3(ZrF6)]\u00b7H2O and crystallizes in the monoclinic space group P21/n to 2.0430\u2005(6)\u2005\u00c5. The \u039b-shape, indicated by the Cu1\u2014F1\u2014Zr1 bond angle of 132.59\u2005(3)\u00b0, is enforced by an intra\u00admolecular O2\u2014H2B\u22efF6 hydrogen bond (Table\u00a0262\u2212 group significantly relative to the TiF62\u2212 group in compound (I)Compound (II)/n Fig.\u00a02. The strd Table\u00a02. The sin2O)3(HfF6)]\u00b7H2O and crystallizes in the monoclinic space group P21/n /n Fig.\u00a03. Compoun2O)2F]2[HfF6]\u00b7H2O and crystallizes in the monoclinic space group P21/n 2F+ cations and octa\u00adhedral HfF62\u2212 anions. The free HfF62\u2212 octa\u00adhedron occupies an inversion center with three distinct bond lengths ranging between 1.9863\u2005(10) and 1.9957\u2005(9)\u2005\u00c5.Compound (IV)/n Fig.\u00a04. The strThe \u039b-shaped building units in compounds (I)\u2013(III) are arranged in head-to-tail chains via inter\u00admolecular hydrogen bonding with multiple hydrogen-bonding inter\u00adactions and \u03c0\u2013\u03c0 stacking contacts to adjacent chains.2O)3(TiF6)] complex in compound (I)2O)3(TiF6)] complexes and three free water mol\u00adecules 3(MF6)] units in compound (II)2O)3(MF6)] complexes and three contacts to hydrating water mol\u00adecules 3(MF6)] complexes participate in parallel displaced \u03c0\u2013\u03c0 stacking inter\u00adactions (Table\u00a05The [Cu(phen)2F+ complexes (H2O)2F+ complexes and O1\u2014H1B\u22efF4 hydrogen bonds with HfF62\u2212 groups (Table\u00a04B\u22efF1 hydrogen bond to an adjacent Cu(phen)(H2O)2F+ complex and a O2\u2014H2A\u22efO3 hydrogen bond with a free water mol\u00adecule (Table\u00a04MF62\u2212 group forms hydrogen bonds with four free water mol\u00adecules and two Cu(phen)(H2O)2F+ complexes. The Cu(phen)(H2O)2F+ complexes pack with both face-to-face and parallel displaced \u03c0\u2013\u03c0 stacking inter\u00adactions (Table\u00a05In compound (IV)es Fig.\u00a08. The equs Table\u00a04. The apie Table\u00a04. Each MFs Table\u00a05.2O)5(VO(H2O)F4)]\u00b7H2O 5(VOF4(H2O))]\u00b7H2O contains a mol\u00adecular \u039b-shaped [Cu(H2O)5(VOF4(H2O))] mol\u00adecule that is bridged via the Cu1\u2014O8\u2014V1 linkage with a bond angle of 142.88\u00b0. The \u039b-shape of this complex is supported by a single intra\u00admolecular hydrogen bond as well as two hydrogen-bonding inter\u00adactions with a free water mol\u00adecule that serves as an inter\u00admolecular \u2018bridging mol\u00adecule\u2019. In contrast, the hydrating water mol\u00adecules in compounds (I)2O)5(VOF4(H2O))]\u00b7H2O is 88.42\u00b0, meaning that the complex has a small tilt similar to compound (I)Aside from compounds (I)2O)5(VO(H2O)F4)]\u00b7H2O are arranged in a polar NCS lattice containing head-to-head/tail-to-tail chains in which the polar moments of the \u039b-shaped complexes are partially aligned perpendicular to the chain direction, with head-to-tail orientations between chains. In contrast, the \u039b-shapes found in compounds (I)The \u039b-shapes in [Cu2, 2.56\u2005mmol of 1,10-phenanthroline, 1.0\u2005mL (27.6\u2005mmol) of HF(aq) (48%), and 0.1\u2005mL (5.5\u2005mmol) of deionized H2O. Compound (II)2, 2.56\u2005mmol of phen, 1.0\u2005mL (27.6\u2005mmol) of HF(aq) (48%), and 0.1\u2005mL (5.5\u2005mmol) of deionized H2O. Compound (III)2, 2.56\u2005mmol of phen, 1.0\u2005mL (27.6\u2005mmol) of HF(aq) (48%), and 0.1\u2005mL (5.5\u2005mmol) of deionized H2O. Compound (IV)2, 2.56\u2005mmol of phen, 0.4\u2005mL (11.03\u2005mmol) of HF(aq) (48%), and 0.7\u2005mL (38.85\u2005mmol) of deionized H2O.The compounds reported here were synthesized by the hydro\u00adthermal pouch method (Harrison Uiso(H) = 1.2Ueq(C) within OLEX2 .Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989021000645/tx2034sup1.cifCrystal structure: contains datablock(s) I, II, III, IV, I, II, III, IV. DOI: 10.1107/S2056989021000645/tx2034Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989021000645/tx2034IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989021000645/tx2034IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 10.1107/S2056989021000645/tx2034IVsup5.hklStructure factors: contains datablock(s) IV. DOI: 2045776, 2045775, 2045774, 2045773CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Retinal ischemia is a medical condition associated with numerous retinal vascular disorders, such as age-related macular degeneration, glaucoma, and diabetic retinopathy. This in vitro cell and in vivo animal study investigated not only the protective effect of S-allyl L-cysteine against retinal ischemia but also its associated protective mechanisms. \u03b1, and vascular endothelium factor (VEGF) in the proposed anti-ischemic mechanism. Lastly, the safety of drug consumption was investigated for changes in the animal's body weight, ERG waves, and blood biochemical parameters . Retinal ischemia was mimicked by raising the intraocular pressure to 120\u2009mmHg for 1 hour in one eye. The effects of pre-/postischemic administration of vehicle vs. SAC 0.18\u2009mg vs. SAC 0.018\u2009mg vs. SAC 0.0018\u2009mg treatments on retina cells were evaluated through cellular viability (MTT assay), flash electroretinograms (ERGs), and fluorogold retrograde labelling counting). Also, protein immunoblot was utilized to assess the role of Wnt, hypoxia inducible factor (HIF)-1\u03b1/VEGF protein was ameliorated significantly by preischemic low dose of SAC. In terms of the animal safety, no significant body weight and electrophysiological differences were observed among defined different concentrations of SAC without following ischemia. In low SAC dosage and vehicle groups, various blood biochemical parameters were normal; however, high and medium concentrations of SAC significantly lowered the levels of uric acid, Hb, and MCHC. The characteristic ischemic changes including significant reduction in ERG b-wave ratio and RGC number were significantly counteracted by pre- and postischemic low dose of SAC. Additionally, ischemia-induced overexpression of Wnt/HIF-1\u03b1/VEGF might presently explain SAC's anti-ischemic mechanism. This study shows that preischemic administration of low SAC dosage has been proved to be safe and most effective against rat retinal ischemia electrophysiologically and/or histopathologically. Moreover, counteracting the ischemia-induced overexpression of Wnt/HIF-1 Common retinal ischemic disorders include central/branch retinal artery occlusion, central/branch retinal vein occlusion, glaucoma, diabetic retinopathy, and neovascular age-related macular degeneration. These diseases are associated with visual impairment and even blindness in more severe cases. Characteristic retinal ischemic findings are known to affect electroretinogram's b-wave amplitude and retinal ganglion cell (RGC) number . There a\u03b2-catenin expression. This is done by binding to frizzled and low-density lipoprotein receptor-related protein 5 or 6, which prevents the breakdown of \u03b2-catenin [\u03b2-catenin can work with lymphoid enhancer-binding factor 1, in order to regulate the target gene expression as vascular endothelial growth factor (VEGF) [\u03b1 (HIF-1\u03b1), which consequently also increases VEGF levels in hypoxia [The current knowledge is that the canonical Wnt pathway is said to stabilize -catenin . Liu et r (VEGF) . Other s hypoxia \u201312. Our hypoxia .\u22128\u2009g/kg\u22483\u223c150\u2009ng of SAC has been proven to protect the human eye against retinal ischemia via its antioxidative and other neuroprotective properties [Therefore, it is important to carry out an investigation into the ischemic alterations associated with the Wnt pathway as well as provide an alternative and complementary treatment to it. Due to the limitations of current treatment , new strategies for retinal vascular disorders driven by persistent ischemia/hypoxia are needed . In thisoperties , 8. Lastoperties makes it\u03b1, or VEGF (identified through western blotting assay). Finally, the safety of fortified SAC in animals was also evaluated through its effects on the animal's body weight (BW), electrophysiology, and biochemical alterations. Overall, it is hypothesized that SAC would be able to nullify ischemic-induced insults in the retina and with little side effects. Specifically, the addition of SAC would downregulate the level of Wnt, HIF-1\u03b1, and VEGF protein expression. Thus, it would lead to less pronounced retinal ischemia associated alterations, namely, decreased RGC number and ERG b-wave amplitude.In the following study, the effects and mechanisms of SAC against retinal ischemia-reperfusion (I/R) injury were explored, via cellular viability analysis, electroretinogram measurement along with retrograde fluorogold labelling RGC number count, and also its regulation on the defined proteins of Wnt, HIF-1\u03bcg (0.00018\u2009mg) in a 0.2\u2009kg rat and 10\u2009\u03bcg of a 70\u2009kg human. This means that the cellular concentration of the fortified (10-fold) SAC is equivalent to 0.0018\u2009mg (1.8\u2009\u03bcg) per day in animal tests. In the present in vitro cellular viability tests, ten (1\u2009mM), hundred (10\u2009mM), and thousand folds (100\u2009mM) of the original dose were prepared. These are equivalent to 0.0018 (low dose), 0.018 (medium dose), and 0.18\u2009mg (high dose) used in the in vivo animal studies.SAC was purchased from Sigma-Aldrich and prepared by dissolving it in distilled water. Based on our previous publication , the maxFor the animal study, defined administration by oral gavage of daily dose of SAC was carried out in these following groups, namely, a preischemic administration (SAC 0.0018\u2009mg\u2009+\u2009I/R and SAC 0.018\u2009mg\u2009+\u2009I/R) for 4 weeks or a postischemic administration (I/R\u2009+\u2009SAC 0.0018\u2009mg) for seven days. In contrast, the rats in the Vehicle\u2009+\u2009I/R group subjected to ischemia were preadministrated with the same volume of vehicle as that of the pre-/postadministrated SAC.The RGC-5 cell lines for the dosing regimen were prepared using the protocol described by Chao et al. but with\u03bcL of experimental cells and allowed to react for a total of 3\u2009h at 37\u00b0C. Then, the reduced MTT was solubilized by the addition of 100\u2009\u03bcL of dimethyl sulfoxide [The MTT assay was carried out, in order to test the efficient concentrations of SAC. When the NAD(P) H-dependent cellular oxidoreductase enzymes reduces MTT, it forms a purple-colored substance called formazan. In this case, a more purple solution reflects a higher number of viable cells. In order to conduct this test, MTT was placed into 96-well plate that contains 100\u2009This animal study followed the regulations, which is set by ARVO Statement for the Use of Animals in Ophthalmology and Vision Research. Also, the permission to conduct the study was obtained from institutional review board of Cheng-Hsin General Hospital . As for the animals, six-week-old Wistar rats (BioLasco) were purchased and reared in an environment set at 40\u223c60% humidity and 19\u223c23\u00b0C temperature. Throughout the experimental days, the animals were kept on a 12\u2009h light/dark setting along with 12\u223c15 air refreshment per hour and also given food and water at their own pleasure.Then, the animals were randomly placed into normal, ischemic (Vehicle\u2009+\u2009I/R), preischemic treatment groups (SAC 0.0018\u2009mg\u2009+\u2009I/R and SAC 0.018\u2009mg\u2009+\u2009I/R) and postischemic treatment group (SAC I/R\u2009+\u20090.0018\u2009mg).An intraperitoneal injection was employed to anesthetize the rats. Specifically, 100\u2009mg/kg ketamine (Pfizer) and 5\u2009mg/kg xylazine (Sigma-Aldrich) are given to the rat, which causes the effect of sedation along with analgesia. Another intraperitoneal injection of 140\u2009mg/kg sodium pentobarbital (SCI Pharmtech) was conducted to humanely kill the rat (Scientific Procedures Acts 1986).Each rat (200\u2013250\u2009g) was anesthetized using the method described above and set onto a stereotaxic frame. As described in Chao et al. , a rat mAfter the rats were euthanized, blood sample is slowly taken from the rat's left ventricle, via technique known as cardiac puncture . Then, t\u22121 was given via a strobe 2\u2009cm before the rat's eyes. In this case, fifteen continuous recordings were done at each 2\u2009s interval, which is set at 10\u2009s\u22121. Afterwards, their amplitudes were maximized and calibrated to give a mean, via usage of an amplifier P511, regulated power supply RPS107, and stimulator PS22 (Grass-Telefactor). The b-wave amplitude ratio of I/R eye to that of the normal eye was measured, in order to compare between different experimental treatments as illustrated by Chao et al. [For the flash ERG measurements, they were carried out on day 0 (nonadministered rats) along with the day after vehicle\u2009+\u2009I/R, SAC\u2009+\u2009I/R, or I/R\u2009+\u2009SAC procedure. Then, the dark adaptation was carried out for a total of 8 hours, and anesthesia was conducted for ERG recording with pupil dilation. Then, a stimulus of 0.5\u2009so et al. .\u03bcl of 5% fluorogold (Sigma-Aldrich) at 3.8, 4.0, and 4.2\u2009mm below its skull. Of note, this step was carried out 3 days prior euthanasia of rat. Then, the retina was dissected and collected, as described by Chao et al. [The anesthetic rat is placed onto the stereotaxic frame, and a 2\u2009cm deep cut is created on the rat's scalp and 2 small holes drilled into its skull . Afterwao et al. . Finallyo et al. .\u03bcg/30\u2009\u03bcl/well, and underwent separation through 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis . Once that is completed, the samples were transferred from the gel onto polyvinylidene fluoride membrane. The membranes itself were soaked with 5% fat-free skimmed milk at 4\u00b0C and blocking buffer for 16 hours. Then, the membranes were incubated with a series of primary antibodies at 25\u00b0C for 1\u2009hour, namely, mouse anti-\u03b2-actin monoclonal antibody , rabbit anti-Wnt3a monoclonal antibody , mouse anti-HIF-1\u03b1 antibody , and rabbit polyclonal anti-VEGF antibody . The blots were then incubated with the relevant secondary antibodies, which were horseradish peroxidase-conjugated goat anti-rabbit IgG and anti-mouse IgG at 25\u00b0C for 1 hour. Lastly, the blots were developed through enhanced chemiluminescent analysis system (HyCell) and exposed to an X-ray film (Fujifilm), and then the proteins levels were analyzed through scanning densitometry.The samples of the retina were isolated and sonicated in lysis buffer after the rat is euthanized. Then, they were divided to similar amount of 30\u2009t test was utilized to compare between two groups, whereas one-way analysis of variance (ANOVA) was carried out for comparison of three or more than 3 independent groups. After the ANOVA tests, Dunnet's test was selected to compare the control versus the rest of the groups . Of note, results were presented as mean\u2009\u00b1\u2009SE, and a P value that is smaller than 0.05 was considered as significant.An unpaired To test the optimal and safe cellular concentration of fortified SAC onOGD-insulted cells, lower and higher concentrations of pre-OGD administration of SAC were tested and then compared against the control group .As demonstrated in P < 0.001; at 1\u2009mM) attenuation of the OGD-induced cellular injury.In contrast, as shown in Overall, the result implies that SAC with concentrations below 1\u2009mM or 0.0018\u2009mg/day should have some form of protective effect against ischemic injury. Thus, this data along with ERG's results (mentioned later) lead to the selection of SAC 0.0018\u2009mg for testing in animal studies, namely, fluorogold labelling and western blot analysis.SAC 0.0018\u2009mg\u201d, which is as follows: baseline (220.2\u2009\u00b1\u200918.1 vs. 217.0\u2009\u00b1\u200918.2 vs. 221.8\u2009\u00b1\u200916.9 vs. 224.0\u2009\u00b1\u200918.1), week 1 (258.2\u2009\u00b1\u200916.0 vs. 257.3\u2009\u00b1\u200914.5 vs. 262.2\u2009\u00b1\u200915.8 vs. 269.5\u2009\u00b1\u200918.2), week 2 (282.2\u2009\u00b1\u200913.8 vs. 280.0\u2009\u00b1\u20098.3 vs. 304.5\u2009\u00b1\u200914.7 vs. 292.0\u2009\u00b1\u200912.1), week 3 (324.0\u2009\u00b1\u200912.3 vs. 310.7\u2009\u00b1\u20096.7 vs. 325.7\u2009\u00b1\u20099.1 vs. 328.2\u2009\u00b1\u20098.2), and week 4 (321.0\u2009\u00b1\u20099.5 vs. 322.3\u2009\u00b1\u20098.7 vs. 323.3\u2009\u00b1\u200912.5 vs. 330.7\u2009\u00b1\u200917.7). Of note, these BW results were represented as mean\u2009\u00b1\u2009SE (g). From the values above, it could be implied that the animals followed a normal growth rate over time (increase of BW), which is not affected by the presence of vehicle or SAC.As shown in SAC 0.0018\u2009mg\u201d across weeks, which enables a comparison between different groups. The ERG a-wave amplitudes in 0.07\u2009\u00b1\u20090.02), week 1 (0.10\u2009\u00b1\u20090.03 vs. 0.09\u2009\u00b1\u20090.02 vs. 0.07\u2009\u00b1\u20090.02 vs. 0.05\u2009\u00b1\u20090.01), week 2 (0.09\u2009\u00b1\u20090.03 vs. 0.08\u2009\u00b1\u20090.02 vs. 0.10\u2009\u00b1\u20090.02 vs. 0.06\u2009\u00b1\u20090.02), week 3 (0.07\u2009\u00b1\u20090.02 vs. 0.07\u2009\u00b1\u20090.02 vs. 0.06\u2009\u00b1\u20090.02 vs. 0.06\u2009\u00b1\u20090.02), and week 4 (0.09\u2009\u00b1\u20090.03 vs. 0.07\u2009\u00b1\u20090.02 vs. 0.09\u2009\u00b1\u20090.02 vs. 0.06\u2009\u00b1\u20090.01). Throughout the 4 weeks of observation, there was no significant difference of a-wave amplitudes between each group and at each week.By means of ERG, preischemia daily doses of vehicle or SAC have been proved to be safe, as shown in Tables 0.41\u2009\u00b1\u20090.07), week 1 (0.49\u2009\u00b1\u20090.05 vs. 0.44\u2009\u00b1\u20090.05 vs. 0.44\u2009\u00b1\u20090.06 vs. 0.45\u2009\u00b1\u20090.06), week 2 (0.43\u2009\u00b1\u20090.07 vs. 0.37\u2009\u00b1\u20090.06 vs. 0.46\u2009\u00b1\u20090.01 vs. 0.47\u2009\u00b1\u20090.08), week 3 (0.41\u2009\u00b1\u20090.07 vs. 0.35\u2009\u00b1\u20090.04 vs. 0.44\u2009\u00b1\u20090.06 vs. 0.40\u2009\u00b1\u20090.01), and week 4 (0.35\u2009\u00b1\u20090.04 vs. 0.38\u2009\u00b1\u20090.05 vs. 0.44\u2009\u00b1\u20090.03 vs. 0.41\u2009\u00b1\u20090.04). Similarly, these data are also not significantly different across different groups and various weeks.As for Of note, the ERG a-wave amplitudes are relatively smaller and sometimes harder to be measured than b-wave amplitudes. Thus, it is usually disregarded for the test of therapeutic properties of drugs; instead, it is presently for safety tests and evidence of retinal normality.https://pubmed.ncbi.nlm.nih.gov/31651183/ [SAC 0.0018\u2009mg\u201d for comparison.As reported in previous publication 1651183/ , blood ln\u2009=\u20094\u223c11): for glucose (136.55\u2009\u00b1\u20094.67 vs. 124.71\u2009\u00b1\u20096.83 vs. 128.76\u2009\u00b1\u20093.73 vs. 130.39\u2009\u00b1\u20098.41), blood urea nitrogen (15.89\u2009\u00b1\u20090.51 vs. 15.39\u2009\u00b1\u20090.42 vs. 15.75\u2009\u00b1\u20090.65 vs.17.14\u2009\u00b1\u20091.25), creatinine (0.41\u2009\u00b1\u20090.01 vs. 0.43\u2009\u00b1\u20090.01 vs. 0.42\u2009\u00b1\u20090.02 vs. 0.47\u2009\u00b1\u20090.02), AST (109.71\u2009\u00b1\u20095.91 vs. 104.25\u2009\u00b1\u20095.46 vs. 112.86\u2009\u00b1\u20099.12 vs. 107.71\u2009\u00b1\u20092.30), ALT (35.67\u2009\u00b1\u20091.62 vs. 31.43\u2009\u00b1\u20090.84 vs. 33.71\u2009\u00b1\u20092.16 vs. 34.86\u2009\u00b1\u20091.72), ALKP (101.40\u2009\u00b1\u20095.22 vs. 113.00\u2009\u00b1\u20094.99 vs. 113.17\u2009\u00b1\u20095.38 vs. 108.14\u2009\u00b1\u20093.94), triglyceride (39.11\u2009\u00b1\u20096.08 vs. 35.57\u2009\u00b1\u20093.47 vs. 38.00\u2009\u00b1\u20097.48 vs. 35.22\u2009\u00b1\u20093.78), cholesterol (41.89\u2009\u00b1\u20092.34 vs. 38.11\u2009\u00b1\u20093.01 vs. 36.00\u2009\u00b1\u20092.16 vs. 40.33\u2009\u00b1\u20091.35), high-density lipoprotein , low-density lipoprotein , red blood cells (8.12\u2009\u00b1\u20090.16 vs. 8.00\u2009\u00b1\u20090.11 vs. 7.79\u2009\u00b1\u20090.18 vs. 8.19\u2009\u00b1\u20090.13), hematocrit (46.03\u2009\u00b1\u20090.65 vs. 44.04\u2009\u00b1\u20090.49 vs. 44.94\u2009\u00b1\u20090.77 vs. 44.24\u2009\u00b1\u20090.60), mean corpuscular volume (MCV: 55.68\u2009\u00b1\u20090.72 vs. 55.46\u2009\u00b1\u20090.50 vs. 55.46\u2009\u00b1\u20090.85 vs. 54.51\u2009\u00b1\u20090.67), mean corpuscular hemoglobin , and platelet (1018.89\u2009\u00b1\u200933.50 vs. 1022.00\u2009\u00b1\u200936.15 vs. 1148.50\u2009\u00b1\u200946.56 vs. 1032.50\u2009\u00b1\u200969.83). In addition, there were also no significant differences among various groups (n\u2009=\u20096\u223c8) for albumin (1.51\u2009\u00b1\u20090.04 vs. 1.68\u2009\u00b1\u20090.04 vs. 1.58\u2009\u00b1\u20090.03) and WBC (6.19\u2009\u00b1\u20090.29 vs. 7.140.39 vs. 6.80\u2009\u00b1\u20090.51).After 4 weeks of treatment, there were no significant differences among various groups (P=0.005) vs. 1.12\u2009\u00b1\u20090.10 (P < 0.001) vs. 1.49\u2009\u00b1\u20090.23. As for P < 0.001) vs. 15.81\u2009\u00b1\u20090.26 vs. 15.41\u2009\u00b1\u20090.22. Finally, the mean corpuscular hemoglobin concentration found in P=0.006) vs. 34.73\u2009\u00b1\u20090.18 vs. 34.15\u2009\u00b1\u20090.35.Of note, the only significant side effects were the following parameters displayed in P < 0.001) resulted in ERG b-wave ratio reduction of 0.20\u2009\u00b1\u20090.04 (n\u2009=\u20098), relative to that of normal (control) retina . The ischemia-induced ERG b-wave reduction was either tended to be alleviated or significantly blunted by pre- or postischemia oral ingestion of SAC, namely, SAC 0.018\u2009mg\u2009+\u2009I/R , SAC 0.0018\u2009mg\u2009+\u2009I/R , and I/R\u2009+\u2009SAC 0.0018\u2009mg .As shown in Figures n\u2009=\u20094), the Vehicle\u2009+\u2009I/R treatment led to significant (P < 0.001) reductions in RGC numbers per field of 185.48\u2009\u00b1\u20097.50 elevated number of RGCs cells, namely, 329.52\u2009\u00b1\u200917.25\u2009cells/field for SAC 0.0018\u2009mg\u2009+\u2009I/R upregulation of target proteins, namely, Wnt3a are target genes of Wnt pathway, which largely correlated with the development of ischemic-related ocular diseases [The results show that I/R significantly affected the retina in a number of ways, via cellular viability Figures and 4, ediseases .\u03b1 and in return increases VEGF [\u03b1 for 4 weeks was proven to be the safest and most effective, via observation of body weight changes , ERG alt\u03b1/VEGF overexpression , relative to conventional treatment methods such as anti-VEGF.Preischemia administration of fortified SAC (low dose: 0.0018\u2009mg) is proven to be experimentally safe as shown by the animal safety experiments and electrophysiological/histopathological data, in terms of the ability of SAC to alleviate retinal ischemia injury. Of medical importance and novelty to this paper, the ischemia-induced elevation of upstream protein Wnt3a and its associated downstream target HIF-1"} +{"text": "Status epilepticus (SE) is a life-threatening neurological disorder. The hippocampus, as an important area of the brain that regulates cognitive function, is usually damaged after SE, and cognitive deficits often result from hippocampal neurons lost after SE. Fyn, a non-receptor Src family of tyrosine kinases, is potentially associated with the onset of seizure. Saracatinib, a Fyn inhibitor, suppresses epileptogenesis and reduces epileptiform spikes. However, whether saracatinib inhibits cognitive deficits after SE is still unknown.In the present study, a pilocarpine-induced SE mouse model was used to answer this question by using the Morris water maze and normal object recognition behavioral tests.We found that saracatinib inhibited the loss in cognitive function following SE. Furthermore, we found that the number of hippocampal neurons in the saracatinib treatment group was increased, when compared to the SE group.These results showed that saracatinib can improve cognitive functions by reducing the loss of hippocampal neurons after SE, suggesting that Fyn dysfunction is involved in cognitive deficits after SE, and that the inhibition of Fyn is a possible treatment to improve cognitive function in SE patients. Epilepsy is a chronic neurological disorder characterized by a persistent occurrence of seizures. Status epilepticus (SE) is characterized by prolonged seizures or intermittent seizures and unconsciousness , which pA receptors and is associated with both excitatory and inhibitory ion channels, which are potentially associated with the onset of seizure .Sarscatinib was administered (25\u00a0mg/kg) orally starting 2\u00a0h after diazepam injection and repeated twice daily for the first 3\u00a0days followed by a single dose each day for the next 11\u00a0days during the 2\u00a0weeks after SE. Same volume of saline was gave to CON mice.Mice that received continuous video-EEG monitoring were implanted with silver wire electrodes (0.125\u00a0mm in diameter) into the hippocampal dentate gyrus (DG) region after anesthesia with pentobarbital . The electrode implantation site used the following coordinates with the bregma as the reference: bregma: \u2212\u20092.3\u00a0mm, lateral: 1.8\u00a0mm and depth: 2.0\u00a0mm. The reference electrode was placed in the frontal cortex. All implanted surgery was performed at 7\u00a0days before the induced SE. EEG activity was recorded 12\u00a0h every day for up to 7\u00a0days at 14\u00a0days after induced SE using PowerLab 8/35 software . Epileptic spikes (sharp (<\u200950\u00a0ms) positive or negative deflections with amplitudes exceeding twice the baseline EEG) were detected and scored by the Gotman spike using PowerLab software. Mice behavior was monitored using video and reviewed by an investigator who was blinded to the identity of the groups (n\u2009=\u20095 each in every group).One month after induced SE, the mice were anesthetized with pentobarbital and transcardially perfused with phosphate-buffered saline (PBS) followed by 4% paraformaldehyde in 0.1\u00a0M PBS to sacrifice the mice. The brain was dissected and fixed in 4% paraformaldehyde overnight at 4\u00a0\u00b0C and then cryoprotected in 30% sucrose in PBS for 72\u00a0h at 4\u00a0\u00b0C. Coronary sections\u00a0(8 \u03bcm) were cut with a cryostat and mounted onto glass slides. Sections were washed in PBS and rehydrated in ethanol of decreasing concentrations. After washing with PBS, the sections were incubated with anti-NeuN antibody overnight at 4\u00a0\u00b0C and washed with PBS. The sections were then incubated with secondary antibody and visualized using 3,3-diaminobenzidine. NeuN-positive neurons in a 100 \u03bcm line long DG of hippocampus were counted in five sections from identical levels in each mouse (n\u2009=\u20095/groups).p\u2009<\u20090.05 was considered statistically significant.Data are expressed as the mean\u2009\u00b1\u2009SEM. One-way or two-way analysis of variance was used to assess differences between two groups and the least significant difference (LSD) or Dunnett T3 post hoc test was used to compare multiple groups after normality tests. A value of Mice often exhibited hypoactivity, curling up, tremors, head bobbing, and myoclonic movement of the limbs after pilocarpine injection. SE usually began by rearing with or without falling and jumping at the onset of SE.t(8)=2.986, p\u2009=0.007). Furthermore, video-EEG showed that saracatinib injection decreased the duration =2.763, p\u2009=0.025) and frequency =3.715, p\u2009=0.006) of spontaneous seizure after SE \u2009=\u20093.68; CON vs SE: 33.59\u2009\u00b1\u20093.8 vs 50.29\u2009\u00b1\u20094.87 on 3\u00a0days, p\u2009=\u20090.13; F\u2009=\u20097.116; CON vs SE vs Sar\u2009+\u2009SE: 22.13\u2009\u00b1\u20093.4 vs 45.83\u2009\u00b1\u20095.8 vs 30.11\u2009\u00b1\u20094.18 on 4\u00a0days, p\u2009=\u20090.01 in CON vs SE, p\u2009=\u20090.025 in SE vs Sar\u2009+\u2009SE; F\u2009=\u200910.307; CON vs SE vs Sar\u2009+\u2009SE: 17.42\u2009\u00b1\u20092.88 vs 41.72\u2009\u00b1\u20093.94 vs 26.45\u2009\u00b1\u20094.69 on 5\u00a0days, p\u2009=\u20090.009 in CON vs SE; p\u2009=\u20090.012 in SE vs Sar\u2009+\u2009SE) and swimming path length from the starting position to the target platform \u2009=\u20093.02; CON vs SE: 1589.37\u2009\u00b1\u2009213.03 vs 2490.87\u2009\u00b1\u2009296.99 on 3\u00a0days, p\u2009=\u20090.23; F\u2009=\u20098.54; CON vs SE: 1138.7\u2009\u00b1\u2009166.62 vs 2381.65\u2009\u00b1\u2009258.46 on 4\u00a0days, p\u2009=\u20090.15; F\u2009=\u200913.81; CON vs SE vs Sar\u2009+\u2009SE: 802.907\u2009\u00b1\u2009103.21 vs 1786.48\u2009\u00b1\u2009155.98 vs 1070.8637\u2009\u00b1\u2009113.52 on 5\u00a0days, p\u2009=\u20090.001 CON vs SE; p\u2009=\u20090.16 SE vs Sar\u2009+\u2009SE) were decreased in Sar\u2009+\u2009SE mice, when compared with the SE mice. However, the swimming speed was similar in all groups mice \u2009=\u20090.54; CON vs SE vs Sar\u2009+\u2009SE: 62.09\u2009\u00b1\u20092.69 vs 61.72\u2009\u00b1\u20093.02 vs 57.85\u2009\u00b1\u20093.79 on 3\u00a0days, p\u2009>\u20090.05; F\u2009=\u20090.475, CON vs SE vs Sar\u2009+\u2009SE: 57.46\u2009\u00b1\u20092.87 vs 55.94\u2009\u00b1\u20093.3 vs 59.62\u2009\u00b1\u20092.6 on 4\u00a0days, p\u2009>\u20090.05; F\u2009=\u20090.11, CON vs SE vs Sar\u2009+\u2009SE: 59.01\u2009\u00b1\u20092.36 vs 57.85\u2009\u00b1\u20092.7 vs 57.43\u2009\u00b1\u20092.46 on 5\u00a0days, p\u2009>\u20090.05). There results suggested that decreased escape latency and swimming path length in the Sar\u2009+\u2009SE group mice were not due to an impaired capacity of swim. When the target platform was removed in the probe trails, the number of crossing target platform areas \u2009=\u20095.917; CON vs SE vs Sar\u2009+\u2009SE: 3.22\u2009\u00b1\u20090.43 vs 1.25\u2009\u00b1\u20090.25 vs 2.5\u2009\u00b1\u20090.5, p\u2009=\u20090.012 in CON vs SE; p\u2009=\u20090.04 in SE vs Sar\u2009+\u2009SE) and staying time in the quadrant \u2009=\u20095.202; CON vs SE vs Sar\u2009+\u2009SE: 20.8\u2009\u00b1\u20091.97 vs 12.84\u2009\u00b1\u20092.14 vs 18.237\u2009\u00b1\u20090.922, p\u2009=\u20090.01 in CON vs SE; p\u2009=\u20090.38 in SE vs Sar\u2009+\u2009SE) where the target platform was previously located were increased in the Sar\u2009+\u2009SE group, when compared with the SE group. Furthermore, when we performed the NOR test, the PI was similar in both SE group mice and Sar\u2009+\u2009SE group mice \u2009=\u20090.326; CON vs SE vs Sar\u2009+\u2009SE: 0.58\u2009\u00b1\u20090.017 vs 0.553\u2009\u00b1\u20090.02 vs 0.545\u2009\u00b1\u20090.049, p\u2009>\u20090.05), but the RI of the Sar\u2009+\u2009SE mice was increased \u2009=\u20096.944; CON vs SE vs Sar\u2009+\u2009SE: 0.64\u2009\u00b1\u20090.029 vs 0.453\u2009\u00b1\u20090.039 vs 0.578\u2009\u00b1\u20090.036, p\u2009=\u20090.015 in CON vs SE; p\u2009=\u20090.023 in SE vs Sar\u2009+\u2009SE). Together, these results showed that saracatinib inhibited deficits in cognitive function after SE.Cognitive function deficits are a common sequelae after SE, so we examined whether saracatinib inhibited deficits in cognitive function after SE. Using the MWM test, escape latency Fig.\u00a0a \u2009=\u200912.32; CON vs SE vs Sar\u2009+\u2009SE: 30.32\u2009\u00b1\u20091.65 vs 20.48\u2009\u00b1\u20091.09 vs 26.44\u2009\u00b1\u20091.42, p\u2009=\u20090.005 in CON vs SE; p\u2009=\u20090.032 in SE vs Sar\u2009+\u2009SE), suggesting that saracatinib relieved the cognitive function deficits by rescuing hippocampal neuronal loss during SE.Because hippocampal neuron loss is an important reason for the deficits in cognitive function after SE, we next determined the number of hippocampal neurons. Neurons in the DG region in the hippocampus were visualized by anti-NeuN antibody using immunohistochemical staining Fig.\u00a0a. The reSE is a neurological condition with high mortality, involving continuous seizure activity, which is often considered as one of the precipitating factors for temporal lobe epilepsy (TLE). The hippocampus and piriform cortex are brain areas commonly damaged by SE. In addition, the amygdala, thalamus, neocortex, and cerebellum are also brain areas that are usually damaged after SE . NeuronaThe Src kinase family is a family of non-receptor tyrosine kinases that plays important roles in regulating signal transduction. Fyn, as a member of the Src family kinases, plays critical roles in regulating cognitive function in AD and in frontotemporal dementia (FTD) patients , 26. FynLoss of hippocampal neurons is the most common reason for cognitive deficits. Neuronal loss frequency occurs in both patients with TLE and in animal models of SE , 22, 32.The Fyn inhibitor, saracatinib, decreased cognitive deficits by attenuating the loss of hippocampal neurons in a pilocarpine-induced SE mouse model, indicating that inhibition of Fyn activity can potentially improve cognitive deficits in patients with TLE. However, further investigation is necessary to identify the mechanism responsible for this neuroprotective effects."} +{"text": "Cyclic GMP\u2010AMP synthase (cGAS) is a cytosolic DNA sensor that catalyzes the synthesis of the cyclic GMP\u2010AMP dinucleotide 2\u20323\u2032\u2010cGAMP. 2\u20323\u2032\u2010cGAMP functions as inducer for the production of type I interferons. Derivatives of this important second messenger are highly valuable for pharmaceutical applications. However, the production of these analogues requires complex, multistep syntheses. Herein, human cGAS is shown to react with a series of unnatural nucleotides, thus leading to novel cyclic dinucleotides. Most substrate derivatives with modifications at the nucleobase, ribose, and the \u03b1\u2010thio phosphate were accepted. These results demonstrate the catalytic promiscuity of human cGAS and its utility for the biocatalytic synthesis of cyclic dinucleotide derivatives. Enzymatic shortcut: Cyclic dinucleotides, which are of great interest to study immunology and immune oncology, can be synthesized in a one\u2010step biotransformation significantly shortening the chemical synthesis route. The enzyme displays a surprisingly large substrate scope. Cyclic dinucleotides are second messengers that can be found in prokaryotes and also in eukaryotes.Vibrio cholerae.During recent years, the structure, regulation, mechanism, and kinetics of human cGAS and other homologues have been described.4b, 5 Herein, we show that the substrate promiscuity of human cGAS can be exploited to produce a series of 2\u20323\u2032\u2010cGAMP analogues in a one\u2010step reaction superior to classical synthesis routes. The substrate derivatives were chosen with modifications at different positions in the nucleobase, ribose or \u03b1\u2010phosphate Scheme\u2005. Each su2\u2010ATP (entry\u20053) and ATP\u2010\u03b1\u2010S (entry\u200516) and only minor amounts for the substrate 8\u2010Cl\u2010ATP (entry\u200513). Higher specific activities between 40 and 43\u2005mU\u2009mg\u22121 were determined for the derivatives 8\u2010Br\u2010dATP (entry\u200511), 8\u2010Br\u2010ATP (entry\u200510) and GTP\u2010\u03b1\u2010S (entry\u200517) with conversions between 56 and 93\u2009%. The specific activities are comparable to those measured for the conversion of the natural substrates ATP and GTP (74\u2005mU\u2009mg\u22121). The highest specific activity of 136\u2005mU\u2009mg\u22121 was measured for the product synthesis from 8\u2010Br\u2010GTP with a yield of 85\u2009%. Thus, the addition of a bromide at the 8\u2010position of the nucleobase seems to be preferred for both, ATP and GTP, substrates. Another interesting substrate derivative, the \u03b1\u2010thio phosphate GTP analogue (entry\u200517), was accepted for the cyclization reaction and 93\u2009% was converted within 24\u2005h. This product has relevant chemical properties and is of particular importance, because the phosphothionate diester linkages are more resistant to hydrolysis than the phosphate diesters.The specific activities and conversions within 24\u2005h were calculated for all reactions Table\u2005. No prodThe cGAS products were additionally analyzed by electrospray ionization\u2010liquid chromatography\u2010mass spectrometry (ESI\u2010LC\u2010MS) and the expected masses were compared with the detected product masses. The synthesis of nine of the 17 products was confirmed with ESI\u2010LC\u2010MS analysis were converted into the corresponding cyclic dinucleotide derivative using cGAS within 24\u2005h. The conversion of 5\u2005\u03bcmol ATP and GTP served as a reference reaction. The reactions resulted in 65\u2009% conversion of GTP with 2\u2032\u2010F\u2010ATP, 47\u2009% GTP with 8\u2010Br\u2010ATP, 35\u2009% GTP with 8\u2010Cl\u2010ATP, 55\u2009% GTP with 8\u2010NH2\u2010ATP, and 58\u2009% GTP with 8\u2010N3\u2010ATP. These conversions were similar, but not fully comparable with the biotransformations on analytical scale, which can only be attributed to the change in the reaction scale. HPLC Purification of 2\u20323\u2032\u2010cGAMP yielded 1.1\u2005mg of product. The derivatives were purified by two\u2010step HPLC to give 0.1\u2005mg cyclic GMP\u20102\u2032\u2010F\u2010AMP, and 4.1\u2005mg cyclic GMP\u20108\u2010NH2\u2010AMP. The syntheses of cGAMP, cyclic GMP\u20102\u2032\u2010F\u2010AMP and cyclic GMP\u20108\u2010NH2\u2010AMP were successfully validated by NMR spectroscopy (see the Supporting Information for details). The products cyclic GMP\u20108\u2010Br\u2010AMP, cyclic GMP\u20108\u2010N3\u2010AMP and cyclic GMP\u20108\u2010Cl\u2010AMP were not purified in quantifiable amounts.Based on our findings mg\u2010scale biotransformations of 2\u2032\u2010F\u2010ATP, 8\u2010Br\u2010ATP, 8\u2010Cl\u2010ATP, 8\u2010NHIn summary, we have reported a short and facile enzymatic synthesis of novel unnatural cyclic dinucleotides based on the promiscuous activity of human cGAS. Remarkably, most tested ATP and GTP substrate derivatives with modifications at the nucleobases, riboses and \u03b1\u2010phosphate were converted into cyclic dinucleotides with their natural counterpart.The relevance of these cyclic dinucleotide derivatives was recently demonstrated by investigating the biological activity towards the STING receptor based on the analysis of the structure\u2010activity relationship and mouse model systems.Analytical scale biotransformation: For enzyme assays, 40\u2005mM HEPES, 10\u2005mM MgCl2, pH\u20057.2 was used with 0.1\u2005mg\u2009mL\u22121 Herring testis DNA and 40\u2005\u03bcg\u2009mL\u22121 human cGAS. The reaction was started by adding 0.5\u2005mM substrate derivative and 0.5\u2005mM substrate, either ATP or GTP. The reaction volume was 1\u2005mL. Negative controls were performed analogously without the addition of enzyme. All samples were incubated at 37\u2009\u00b0C and 300\u2005rpm in an orbital shaker for at least 24\u2005h. Reactions were stopped by heating at 95\u2009\u00b0C for 5\u2005min. The samples were analyzed with HPLC and LC\u2010MS. The activities given in Table\u2005Milligram\u2010scale biotransformation: For milligram\u2010scale biotransformation, 20\u2005mL reaction solution was used in shaking flasks. The reaction was started by adding 40\u2005\u03bcg\u2009mL\u22121 human cGAS. The reaction mixture was incubated at 37\u2009\u00b0C and 200\u2005rpm in an orbital shaker for 24\u2005h. The reaction was stopped by heating at 95\u2009\u00b0C for 5\u2005min. The product solutions were frozen and stored at \u221220\u2009\u00b0C.Fractionation of biotransformation products by HPLC: The frozen samples were lyophilized to dryness, subsequently resolved in 1\u2005mL H2O and centrifuged at 21\u2009000\u2005g for 5\u2005min. The supernatant of the product solution was separated and collected as fractions with a LaChrome Elite HPLC system equipped with an ISAspher 100\u20103C18AQ column, 150\u00d73\u2005mm . The column temperature was set to 30\u2009\u00b0C. The flow rate was set to 1\u2005mL min\u22121. The product solution was repetitively injected with volumes of 99.5\u2005\u03bcL. A gradient of 50\u2005mM triethylamine acetate (TEAA) with 3\u2009% v/v acetonitrile (solvent A) and 100\u2009% acetonitrile (solvent B) was used for the chromatography. The solvent gradient for all products was: 0\u201310\u2005min: 0\u2009% B, 10\u201320\u2005min: 0 to 30\u2009% B, 20\u201322\u2005min: 30\u2009% B, 22\u201325\u2005min 30 to 0\u2009% B, 25\u201330\u2005min 0\u2009% B. Fractions were taken: cGAMP: 12\u201313\u2005min; cyclic GMP\u20102\u2032\u2010F\u2010AMP: 11\u201312\u2005min and 17\u201318\u2005min; cyclic GMP\u20108\u2010Br\u2010AMP: 17\u201319\u2005min; cyclic GMP\u20108\u2010Cl\u2010AMP: 16\u201318\u2005min; cyclic GMP\u20108\u2010NH2\u2010AMP: 11\u201313\u2005min; cyclic GMP\u20108\u2010N3\u2010AMP: 16\u201317\u2005min. Elution of compounds was monitored with a UV detector at 254\u2005nm. The collected fractions were lyophilized and product purification was validated with NMR analysis.2\u2010AMP: 0\u201318\u2005min: 0\u2009% B, 18\u201328\u2005min: 0 to 30\u2009% B, 28\u201330\u2005min: 30 to 90\u2009% B, 30\u201338\u2005min: 90\u2009% B, 38\u201340\u2005min: 90 to 0\u2009% B, 40\u201350\u2005min: 0\u2009% B; Fractions were taken: 25.9\u201327.9\u2005min; cyclic GMP\u20108\u2010N3\u2010AMP: 0\u201315\u2005min: 0 to 22.5\u2009% B, 15\u201317\u2005min: 22.5 to 90\u2009% B, 17\u201325\u2005min: 90\u2009% B, 25\u201327\u2005min: 90 to 0\u2009% B, 27\u201337\u2005min: 0\u2009% B; Fractions were taken: 7.1\u20139.1\u2005min. Elution of compounds was monitored with a UV detector at 254\u2005nm. The collected fractions were lyophilized and product purification was validated with NMR analysis.For further purification of the product derivatives, a second fractionation using chromatography was performed. In the second fractionation, the solvent gradient was individually adapted for each compound: cyclic GMP\u20102\u2032\u2010F\u2010AMP: 0\u201320\u2005min: 0 to 22.5\u2009% B, 20\u201322\u2005min: 22.5 to 90\u2009% B, 22\u201330\u2005min: 90\u2009% B, 30\u201332\u2005min 90 to 0\u2009% B, 32\u201342\u2005min 0\u2009% B; Fractions were taken: 8.7\u201310.2\u2005min; cyclic GMP\u20108\u2010Br\u2010AMP: 0\u201320\u2005min: 0 to 15\u2009% B, 20\u201322\u2005min: 15 to 90\u2009% B, 22\u201330\u2005min: 90\u2009% B, 30\u201332\u2005min: 90 to 0\u2009% B, 32\u201342\u2005min: 0\u2009% B; Fractions were taken: 14.3\u201316.3\u2005min; cyclic GMP\u20108\u2010Cl\u2010AMP: 0\u20135\u2005min: 0\u2009% B, 5\u201325\u2005min: 0 to 30\u2009% B, 25\u201327\u2005min: 30 to 90\u2009% B, 27\u201335\u2005min: 90\u2009% B, 35\u201337\u2005min: 90 to 0\u2009% B, 37\u201347: 0\u2009% B; Fractions were taken:12.7\u201314.1\u2005min and 16.3\u201318.3\u2005min; cyclic GMP\u20108\u2010NHNMR analysis: NMR spectra were recorded on a Bruker AV 600 Avance III HD system with D2O as solvent and 25\u2005\u03bcL [D4]methanol as internal standard. The solvent signals were referenced to \u03b4H 3.31\u2005ppm and \u03b4C 49.0\u2005ppm.2\u2010AMPCyclic GMP\u20108\u2010NH: 1H NMR : \u03b4=8.08 , 7.94 , 6.17 , 5.99 , 5.60 , 5.06 , 4.72 , 4.62 , 4.44 , 4.38 , 4.27 , 4.37, 4.14\u2005ppm ; 13C NMR : \u03b4=160.2 (CCO), 154.5 (CN), 153.2 (CN), 153.1 (CN), 150.1 (CH), 149.7 (CN), 142.3 (CH), 118.1 (CN), 117.1 (CN), 91.5 (CH), 86.8 (CH), 84.6 (CH), 80.6 (CH), 75.6 (CH), 74.7 (CH), 72.4 (CH), 71.7 (CH), 66.7 (CH2), 62.9\u2005ppm (CH2).Cyclic GMP\u20108\u2010F\u2019\u2010AMP: 1H NMR : \u03b4=8.30 , 8.29 , 7.87 , 6.47 , 5.96 , 5.67 , 5.56 , 5.12 , 4.60 , 4.57 , 4.40 , 4.25 , 4.16\u2005ppm ; 13C NMR : \u03b4=n.d. (CCO), n.d. (CN), 156.0 (CN), 155.1 (CH), 152.2 (CN), 148.1 (CN), 142.9 (CH), 140.7 (CH), 119.2 (CN), 118.1 (CN), 87.8 (CH), 87.6 (CH), 87.2 (CH), 84.0 (CH), 74.6 (CH), 72.2 (CH), 71.8 (CH) 69.9 (CH), n.d. (CH), 66.8\u2005ppm (CH). (n.d.=not detected)The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "Developing a universal strategy to improve the specificity and sensitivity of PEGylated nanoaparticles (PEG-NPs) for assisting in the diagnosis of tumors is important in multimodality imaging. Here, we developed the anti-methoxypolyethylene glycol (mPEG) bispecific antibody , which has dual specificity for mPEG and human epidermal growth factor receptor 2 (HER2), with a diverse array of PEG-NPs to confer nanoparticles with HER2 specificity and stronger intensity.++) but not MCF7/neo1 cells (HER2+/\u2212). The \u03b1HER2/Lipo-DiR and \u03b1HER2/SPIO could enhance the sensitivity of untargeted PEG-NPs on MCF7/HER2 (HER2++). In in vivo imaging, \u03b1HER2/Lipo-DiR and \u03b1HER2/SPIO increased the specific targeting and enhanced PEG-NPs accumulation at 175% and 187% on 24\u00a0h, respectively, in HER2-overexpressing tumors.We used a one-step formulation to rapidly modify the nanoprobes with mPEG\u2009\u00d7\u2009HER2 and optimized the modified ratio of BsAbs on several PEG-NPs . The \u03b1HER2/PEG-NPs could specifically target MCF7/HER2 cells (HER2mPEG\u2009\u00d7\u2009HER2, therefore, provided a simple one-step formulation to confer HER2-specific targeting and enhanced sensitivity and contrast intensity on HER2 positive tumors for multimodality imaging. Optical imaging (OI) is relatively inexpensive and robust for all kinds of molecular and cellular processes in small animals, but clinical applications are hindered by limited depth penetration /[\u00b5W/cm2].MCF7/HER2 (5\u2009\u00d7\u2009106\u00a0cell/well) were incubated \u03b1HER2/SPIO, \u03b1DNS/SPIO or SPIO were added to the tubes and incubated at 37\u00a0\u00b0C for 1\u00a0h. After washing with PBS three times, cells precipitated at the bottom of the Eppendorf tube and were then imaged with 7T MRI TR/TE, 3000/65\u00a0ms; echo train length, 10; flip angle, 150o; field of view, 6\u00a0cm\u2009\u00d7\u20096\u00a0cm; slice thickness, 1\u00a0mm; interslice gap, 0.1\u00a0mm (10% of slice thickness); and matrix, 192\u2009\u00d7\u2009192. BALB/c nude mice bearing MCF7/HER2 (HER2++) and MCF7/neo1 (HER2+/\u2212) tumor (~\u2009100\u00a0mm3) were injected intravenously with \u03b1HER2/SPIO and \u03b1DNS/SPIO (10\u00a0mg/kg per mouse) in their mammary fat pad regions, respectively. Isoflurane anesthetized mice were imaged with 7T MRI at 0, 24\u00a0h after injection. TR/TE, 3000/65\u00a0ms; echo train length, 10; flip angle, 150; field of view, 4\u00a0cm; slice thickness, 1.2\u00a0mm; interslice gap, 0.12\u00a0mm (10% of slice thickness); and matrix, 256\u2009\u00d7\u2009210. The negative enhancement was calculated by [pretreated signal intensity (SI0)-treated signal intensity (SI24)]/SI0*100.MCF7/HER2 cells and 500\u00a0units/ml recombinant human granulocyte\u2013macrophage colony-stimulating factor (R&D systems), and then seeded 4\u2009\u00d7\u2009105\u00a0cells/well in 24-well plate. On day 6, 20\u00a0ng/ml recombinant human TNF-\u03b1 (Sigma) and 10\u00a0ng/ml IL-1\u03b2 (R&D systems) were added to the cells to activate DCs for 24\u00a0h. On day\u00a07, the harvested DCs were counted and then incubated 50\u00a0\u03bcg/ml mitomycin C for 30\u00a0min at 37\u00a0\u00b0C at a density of 1\u2009\u00d7\u2009106\u00a0cells/ml, then washed extensively. Autologous CD4+ T cells were isolated on Day 7 by negative selection using CD4+ T Cell Isolation Kit II and LS columns (Miltenyi Biotech). After counting, 2\u2009\u00d7\u2009105 CD4+ T Cells were added to 2\u2009\u00d7\u2009104 mitomycin C-treated DCs and incubated with mPEG\u2009\u00d7\u2009HER2, mPEG\u2009\u00d7\u2009DNS at concentration of 350\u00a0nM in 96-well round bottom plates. Controls included dendritic cells plus CD4+ T cells alone and with concentration of 10\u00a0\u03bcg/ml phytohemagglutinin (PHA). Cells were cultured at 37\u00a0\u00b0C for 5\u00a0days. Proliferation was assessed by ATPlite Luminescence Assay kit (Perkin Elmer). Counts per minute (cpm) for each well were determined by multimode plate reader (Perkin Elmer).To prepare monocyte-derived dendritic cells (DCs), peripheral blood mononuclear cells (PBMCs) from healthy donor blood isolated by Ficoll-Paque and monocytes isolated using Miltenyi Pan Monocyte Isolation Kits and LS columns (Miltenyi Biotech). Monocytes were resuspended in RPMI1640 supplemented with 10% FCS, 2\u00a0mM\u00a0Additional file 1: Table S1. The BsAb-conjugation rate of \u03b1HER2/PEG-NPs. Table S2. The Characterization of BsAb/mPEG-NPs. Figure S1. Immunogenicity of humanized BsAbs. We cocultured dendritic cells differentiated from human PBMCs with autologous CD4+ T cells and stimulated with control medium (represented as DC+T), PHA (as positive control), PEG\u00d7HER2, PEG\u00d7DNS, respectively, for 5 days. Then, we detected the proliferation of CD4+ T cells by ATPlite assay. Bars, SD. CPM, counts per minute; PBMC, peripheral blood mononuclear cell; PHA, phytohemagglutinin."} +{"text": "Cordyceps militaris Improves Chronic Kidney Disease by Affecting TLR4/NF-\u03baB Redox Signaling Pathway\u201d [In the article titled \u201cPathway\u201d , there w2)\u201d.\u201cAll the patients were with the CKD late stage 3 or stage 4 (estimated glomerular filtration rate (eGFR) 25 to 40\u2009mL/min)\u201d should read \u201cAll the patients were with the CKD late stage 3 or stage 4 (estimated glomerular filtration rate (eGFR) 25 to 40\u2009mL/min/1.73\u2009m\u201cAll patients met the following criteria: (1) urine\u2009protein/creatinine\u2009ratio < 5\u201d should read \u201cAll patients met the following criteria: (1) urinal\u2009protein > 1\u2009g/24\u2009h.\u201dP < 0\u200905)\u201d should read \u201cAfter the three-month therapy, the values of eGFR (45.6 \u00b1 8.4\u2009mL/min/1.73\u2009m2) were increased significantly when compared with before therapy .\u201d\u201cAfter the three-month therapy, the values of eGFR (28.3 \u00b1 5.2) were reduced significantly when compared with the CG group (32.8 \u00b1 9.2,"} +{"text": "The cyclo\u00adpenta\u00addienyl rings are eclipsed in both of the title compounds and their packing is dominated by H\u22efH contacts, as determined by Hirshfeld surface analyses. N,N-di\u00admethyl\u00adamino)\u00admeth\u00adyl]ferrocene, [Fe(C5H5)(C8H12N)], (1), is an inter\u00adesting starting material for the synthesis of planar chiral 1,2-disubstituted ferrocenes, as demonstrated by the preparation of -bis\u00ad{2-[(di\u00admethyl\u00adamino)\u00admeth\u00adyl]ferrocen\u00adyl}di\u00admethyl\u00adsilane, [Fe2(C5H5)2(C18H18N2Si)], (2), from the li\u00adthia\u00adted derivative of 1. The configuration of the lithium compound is unchanged after the substitution reaction and the chirality is preserved in space group P212121. In both compounds, the Cp rings adopt eclipsed conformations. Hirshfeld surface analysis was used to investigate the inter\u00admolecular inter\u00adactions, and showed that H\u22efH inter\u00adactions dominate in both structures with contact percentages of 83.9 and 88.4% for 1 and 2, respectively.The title compound [( N,N-Di\u00admethyl\u00adamino\u00admethyl\u00adferrocene was first synthesized by Hauser & Lindsay and can be converted by a further step using an electrophile -N,N-dimethyl-1-ferrocenyl\u00adethyl\u00adamine, or Ugi\u2019s amine with a chiral directing group -tetra\u00admethyl-1,2-cyclo\u00adhexa\u00adnedi\u00adamine (TMCDA) with yields in high stereoselectivity and the mechanistic course of the reaction can be described by quantum-chemical calculations -meso-compound of bis\u00ad[dimeth\u00adyl(amino\u00admeth\u00adyl)ferrocen\u00adyl]di\u00admethyl\u00adsilane was characterized by Roewer and co-workers using X-ray diffraction analysis and formed during the synthesis of di\u00admethyldi\u00adchloro\u00adsilane with two equivalents of the racemic li\u00adthia\u00adted N,N-di\u00admethyl\u00adamino\u00admethyl\u00adferrocene -bis\u00ad[dimeth\u00adyl(amino\u00admeth\u00adyl)ferro\u00adcen\u00adyl]di\u00admethyl\u00adsilane (2) and analyze their inter\u00admolecular inter\u00adactions using Hirshfeld surfaces and two-dimensional fingerprint plots.In this paper, we report the crystal structures of 1 crystallizes from n-pentane at 243\u2005K as orange needles with monoclinic (P21/n) symmetry. There are no noticeable irregularities in the bond lengths or bond angles found: the amino\u00admethyl side chain is oriented above its attached cyclo\u00adpenta\u00addienyl ring, and the Cp rings are eclipsed, the dihedral angle between their mean planes being 1.53\u2005(15)\u00b0. The mol\u00adecular structure of 1 is presented in Fig.\u00a01Compound 2 is an orange\u2013red crystalline solid and occurs in enantiomerically pure form in the ortho\u00adrhom\u00adbic space group P212121. The structure is illustrated in Fig.\u00a022 can be assigned the -configuration; furthermore the cyclo\u00adpenta\u00addienyl rings are also in an eclipsed conformation for both iron atoms . The Si\u2014C bonds span the range of 1.869\u2005(3) to 1.874\u2005(3)\u2005\u00c5, which is consistent with the literature \u00b0 (C14\u2014Si1\u2014C10) as the smallest and 112.17\u2005(13)\u00b0 (C14\u2014Si1\u2014C16) as the largest. This flexibility is often observed for Si\u2014C bonds for N2 and 3.584\u2005(3)\u2005\u00c5 for N1 are too long to be regarded as coordinate bonds to Si from the N lone pairs.Compound 1 is shown in Fig.\u00a03dnorm in the range from \u22120.072 to 1.201 (arbitrary units) and the related fingerprint plots generated by CrystalExplorer and the C\u22efH/H\u22efC contacts (11.6%) contribute to the packing arrangement of the crystal. Inter\u00admolecular inter\u00adactions of the cyclo\u00adpenta\u00addienyl rings with neighbouring mol\u00adecules can also be visualized.The crystal packing of compound 1. Selected examples found in the Cambridge Structural Database -meso-bis\u00ad[dimeth\u00adyl(amino\u00admeth\u00adyl)ferrocen\u00adyl]di\u00admethyl\u00adsilane -meso-bis\u00ad[dimeth\u00adyl(amino\u00admeth\u00adyl)ferrocen\u00adyl]di\u00adchloro\u00adsilane -[2-(di\u00admethyl\u00adamino\u00admeth\u00adyl)ferrocen\u00adyl]lithium in n-pentane (1\u2005ml) was made up and stored at 243\u2005K and compound 1 crystallized in the form of orange needles.2 is illustrated in Fig.\u00a07Sp)-[2-(di\u00admethyl\u00adamino\u00admeth\u00adyl)ferrocen\u00adyl]lithium (4.00\u2005mmol) , the product (46%) could be obtained as yellowish plates.The reaction scheme for the synthesis of compound 1H NMR : \u03b4 = 0.81 , 2.02 {s, 12H; [N(CH3)2]2}, 2.80, 3.64 , 4.08 , 4.12 , 4.19 , 4.35 ppm.1H}13C NMR : \u03b4 = 0.6 , 45.4 {4C; [CH2N(CH3)2]2}, 60.5 , 69.7 , 69.8 , 72.6 , 74.2 , 76.6 , 90.5 ppm.{1H}29Si NMR : \u03b4 = \u22127.07 ppm.{m/z (%): 498\u2005(20) [(M\u2013NMe2)+], 409\u2005(100) [(M\u2013NMe2\u2013CH2NMe2\u2013Me2)+], 299\u2005(50) [(M\u2013FcCH2NMe2)+], 199\u2005(50) [(M\u2013SiMe2FcCH2NMe2\u2013NMe2)+].ESI-(+)-MS: Rf: = 0.20.Uiso(H) = 1.2Ueq(C) for CH2 and CH hydrogen atoms and Uiso(H) = 1.5Ueq(C) for CH3 hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2056989020010397/hb7933sup1.cifCrystal structure: contains datablock(s) 1, 2, global. DOI: 10.1107/S2056989020010397/hb79331sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989020010397/hb79332sup3.hklStructure factors: contains datablock(s) 2. DOI: 2019451, 2019450CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title mol\u00adecule contains a 16-membered macrocycle with the conformation of the C\u2014O\u2014C\u2014C\u2014O\u2014C\u2014C\u2014O\u2014C polyether chain being 27H24N2O5, is a product of the deamination reaction from aza-14-crown-4 ether containing the \u03b3-piperidone subunit. The title mol\u00adecule contains a 16-membered macrocycle with the conformation of the C\u2014O\u2014C\u2014C\u2014O\u2014C\u2014C\u2014O\u2014C polyether chain being t\u2013g(-)\u2013t\u2013t\u2013g(+)\u2013t . The dihedral angle between the planes of the benzene rings fused to the aza-14-crown-4-ether moiety is 31.11\u2005(14)\u00b0. The cavity size inside the macrocycle is 4.72\u2005\u00c5. The macrocycle is significantly flattened as a result of the extended conjugated system. Steric repulsion between the pyridyl\u00adcarboxamide fragment and the benzene ring results in a slight deviation of macrocycle from planarity. The structure also features intra\u00admolecular hydrogen bonding, which results in a deviation of the angle between the planes of amide and pyridyl groups from planarity: this angle is 16.32\u2005(18)\u00b0. In the crystal, the mol\u00adecules are linked into infinite zigzag chains via inter\u00admolecular C\u2014H\u22ef\u03c0 contacts. The chains are bound into layers parallel to (100) by weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds.The title compound, C The mol\u00adecular structure of (3) is presented in Fig.\u00a01t\u2013g(-)\u2013t\u2013t\u2013g(+)\u2013t conformation. The dihedral angle between the mean planes of the benzene rings fused to the aza-14-crown-4-ether moiety is 31.11\u2005(14)\u00b0. The cavity size inside the macrocycle, determined as a double-mean distance between the C16, C19, O5, O8 and O11 atoms and the center of this penta\u00adgon, is 4.72\u2005\u00c5. The macrocycle is significantly flattened because of the extended conjugated system. The steric repulsion between the 17-pyridyl\u00adcarboxamide fragment and the aromatic ring (C11A/C12\u2013C15/C15A) results in a slight deviation of the macrocycle from planarity. The mol\u00adecular structure also features intra\u00admolecular hydrogen bonds are linked into infinite zigzag chains via inter\u00admolecular C26\u2014H\u22ef\u03c0(C22) contacts was synthesized according to the procedure described previously and 0.44\u2005g (4.7\u2005mmol) \u03b1-amino\u00adpyridine (2) in 10\u2005ml o-xylene was refluxed with stirring for 5\u2005h (monitored by TLC until the disappearance of the starting organic compound spots). The solvent was evaporated under vacuum, then the residue was purified by column chromatography (ethyl acetate:n-hexane = 5:1) and recrystallized from ethanol to obtain 1.27\u2005g of pure compound (3) as single crystals in 58% yield. Tmlt = 482\u2013484\u2005K. Rf = 0.66 . IR, \u03bd, cm\u22121: 1687 (C=O), 1638 (HN\u2014C=O), 3317 (NH). 1H NMR : 3.89 , 4.01 , 4.33 , 6.91\u20137.75 , 7.73 , 8.18 , 8.32 , 8.36 . Mass spectrum, m/z : 456 [M]+ (4), 428\u2005(1), 309\u2005(4), 283\u2005(3), 265\u2005(3), 238\u2005(25), 221\u2005(18), 210\u2005(50), 189\u2005(10), 173\u2005(89), 159\u2005(20), 147\u2005(38), 131\u2005(100), 118\u2005(48), 115\u2005(51), 103\u2005(27), 91\u2005(81), 89\u2005(52), 78\u2005(65), 45\u2005(38). Analysis calculated for C27H24N2O5, %: C, 71.04; H, 5.30; N, 6.14. Found: C, 70.82; H, 5.34; N, 6.01.A solution of 2.0\u2005g (4.7\u2005mmol) aza-crown ether (Uiso(H) = 1.2Ueq(N)]. The other hydrogen atoms were placed in calculated positions with C\u2014H = 0.95\u20130.99\u2005\u00c5 and refined as riding with fixed isotropic displacement parameters [Uiso(H) = 1.2Ueq(C)].Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020010968/yk2136sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020010968/yk2136Isup2.hklStructure factors: contains datablock(s) I. DOI: 2022314CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are associated into inversion dimers via short Cl\u22efCl contacts [3.3763\u2005(9)\u2005\u00c5]. A Hirshfeld surface analysis indicates that the most important contact percentages for the different types of inter\u00adactions are H\u22efH (43.9%), Cl\u22efH/H\u22efCl (22.9%), C\u22efH/H\u22efC (20.8%) and N\u22efH/H\u22efN (8.0%).In the title compound, C In order to continue our work in this direction, we now describe the synthesis and structure of the title compound, CDatabase survey.The title compound has a non-planar mol\u00adecular conformation Fig.\u00a01; the dihvia short halogen\u22efhalogen contacts compared to the van der Waals radius sum of 3.50\u2005\u00c5. No other directional contacts could be identified and the shortest aromatic-ring-centroid separation is greater than 5.25\u2005\u00c5. The packing for (I)In the crystal, mol\u00adecules of (I)et al., 2007CrystalExplorer17 The Hirshfeld surface (McKinnon a) and those delineated into H\u22efH, Cl\u22efH/H\u22efCl and C\u22efH/H\u22efC contacts . The pair of characteristic wings in the fingerprint plot delineated into C\u22efH/H\u22efC contacts , have the tips at de + di \u2243 2.80\u2005\u00c5. The remaining contributions from the other different inter\u00adatomic contacts to the Hirshfeld surfaces are listed in Table\u00a01et al., 2015The overall two-dimensional fingerprint plot Fig.\u00a04a and thce Fig.\u00a04c. The pet al., 2016E)-1--2-phenyl\u00addiazene unit resulted in 25 hits. Six compounds are closely related to the title compound, viz. 1-(4-bromo\u00adphen\u00adyl)-2-diazene , C\u2014H\u22efCl and C\u2014Cl\u22ef\u03c0 inter\u00adactions into sheets parallel to the ab plane. Additional van der Waals inter\u00adactions consolidate the three-dimensional packing. In the crystal of HODQAV, the planes of the benzene rings make a dihedral angle of 56.13\u2005(13)\u00b0. Mol\u00adecules are stacked in columns along the a-axis direction via weak C\u2014H\u22efCl hydrogen bonds and face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions. The crystal packing is further consolidated by short Cl\u22efCl contacts. In XIZREG, the benzene rings form a dihedral angle of 63.29\u2005(8)\u00b0. Mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds into zigzag chains running along the c-axis direction. The crystal packing also features C\u2014Cl\u22ef\u03c0, C\u2014F\u22ef\u03c0 and N\u2014O\u22ef\u03c0 inter\u00adactions. In the crystals of LEQXIR and LEQXOX, the dihedral angles between the aromatic rings are 56.18\u2005(12) and 60.31\u2005(14)\u00b0, respectively. In LEQXIR, C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and short Cl\u22efO contacts occur and in LEQXOX C\u2014H\u22efN and short Cl\u22efCl contacts are observed.In the crystals of HONBOE and HONBUK, the aromatic rings form dihedral angles of 60.9\u2005(2) and 64.1\u2005(2)\u00b0, respectively. Mol\u00adecules are linked through weak Z)-4-{[2-hydrazineyl\u00adidene]meth\u00adyl}-N,N-di\u00admethyl\u00adaniline , tetra\u00admethyl\u00adethylenedi\u00adamine (TMEDA) , CuCl and CCl4 . After 1\u20133\u2005h the reaction mixture was poured into \u223c0.01 M solution of HCl and extracted with di\u00adchloro\u00admethane (3 \u00d7 20\u2005ml). The combined organic phase was washed with water (3 \u00d7 50\u2005ml), brine (30\u2005ml), dried over anhydrous Na2SO4 and concentrated in vacuo using a rotary evaporator. The residue was purified by column chromatography on silica gel using appropriate mixtures of hexane and di\u00adchloro\u00admethane (3:1\u20131:1) to form a red solid in 85% yield (m.p. 429\u2005K). Orange plates of (I)18H19Cl2N3: C 62.08, H 5.50, N 12.07; found: C 62.01, H 5.48, N 12.03%. 1H NMR \u03b4 2.38 , 3.05 , 6.88\u20137.43 . 13C NMR \u03b4 155.57, 153.15, 151.94, 147.03, 142.69, 138.64, 137.97, 133.14, 131.20, 127.08, 121.02, 21.20. ESI\u2013MS: m/z: 349.18 [M+H]+.A 20\u2005ml screw-neck vial was charged with DMSO (10\u2005ml), (Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C) was applied in all cases.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020009202/hb7912sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020009202/hb7912Isup2.hklStructure factors: contains datablock(s) I. DOI: 2014419CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The phenol ring makes a dihedral angle of 36.56\u2005(3)\u00b0 with the nitro\u00adbenzene ring. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO inter\u00adactions, forming chains along the 15H14N2O3, was prepared by condensation of 2-hy\u00addroxy-5-methyl-benzaldehyde and 2-methyl-3-nitro-phenyl\u00adamine in ethanol. The configuration of the C=N bond is E. An intra\u00admolecular O\u2014H\u22efN hydrogen bond is present, forming an S(6) ring motif and inducing the phenol ring and the Schiff base to be nearly coplanar [C\u2014C\u2014N\u2014C torsion angle of 178.53\u2005(13)\u00b0]. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO inter\u00adactions, forming chains along the b-axis direction. The Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from H\u22efH (37.2%), C\u22efH (30.7%) and O\u22efH (24.9%) inter\u00adactions. The gas phase density functional theory (DFT) optimized structure at the B3LYP/ 6\u2013311\u2005G level is compared to the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.The title compound, C Depending on the tautomers, two types of intra\u00admolecular hydrogen bonds are generally observed in ortho-hy\u00addroxy Schiff bases, namely, O\u2014H\u22efN in enol\u2013imine and N\u2014H\u22efO in keto\u2013amine tautomers ring motif and also induces the phenol ring and the Schiff base to be nearly coplanar, as indicated by the C3\u2014C8\u2014N1\u2014C9 torsion angle of 178.53\u2005(13)\u00b0. An intra\u00admolecular C15\u2014H15B\u22efO2 inter\u00adaction is also observed. The phenol ring (C1\u2013C8/O1) is inclined to the tolyl ring (C9\u2013C14) by 37.57\u2005(3)\u00b0, and the nitro group (N2/O2/O3) is inclined to the tolyl ring (C9\u2014C14) by 35.05\u2005(2)\u00b0. The configuration of the C8=N1 bond is E. The C4\u2014O1 distance is 1.3455\u2005(18)\u2005\u00c5, which is close to normal values reported for single C\u2014O bonds in phenols and salicyl\u00adidene\u00adamines i and C7\u2014H7C\u22efO1i, resulting in the formation of an infinite chain along the b-axis direction . The two-dimensional fingerprint plots provide information about the percentage contributions of the various inter\u00adatomic contacts. The most important are H\u22efH inter\u00adactions, which contribute 37.2% to the total Hirshfeld surface. Other contributions are from C\u22efH (30.7%), O\u22efH (24.9%), N\u22efH (2.0%) and C\u22efO (1.8%) contacts. There are also smaller contributions using the standard B3LYP functional and 6-311G basis-set calculations , the ionization potential (I = \u2212ELUMO), HOMO\u2013LUMO energy gap (\u0394E), the chemical hardness (\u03b7) and softness (S) of the title compound were predicted based on the EHOMO and ELUMO energies. As a result of the large \u0394E and \u03b7 values (Table\u00a03E)-2-{[(3-chloro\u00adphen\u00adyl)imino]\u00admeth\u00adyl}-6-methyl\u00adphenol -2-[(2-hy\u00addroxy-5-meth\u00adoxy\u00adbenzyl\u00adidene)amino]\u00adbenzo\u00adnitrile -4-methyl-2-[(2-methyl-3-nitro-phenyl\u00adimino)\u00admeth\u00adyl]phenol moiety resulted in no hits when both methyl groups were included in the search. Without the methyl groups, seven related compounds were found. Out of these, few are very similar to the title compound and some are metal complexes such as di\u00adazido-}bis\u00ad(4-methyl\u00adpheno\u00adlato)]man\u00adganese amino]-4,5-di\u00adnitro\u00adphen\u00adyl}imino)\u00admeth\u00adyl]-4,6-di-t-butyl\u00adphenolato}iron(III) meth\u00adanol solvate hemihydrate amino]-4,5-di\u00adnitro\u00adphen\u00adyl}imino)\u00admeth\u00adyl]phenolato}iron(III) bis\u00ad]nickel(II) methanol solvate -4,5-di\u00adnitro\u00adphen\u00adyl]aza\u00adnedi\u00adyl}bis\u00ad(methyl\u00adene)]bis\u00ad}meth\u00adano\u00adlcobalt(III) methanol solvate -3-nitro\u00adaniline in ethanol (15\u2005ml) and 2-methyl-3-nitro-phenyl\u00adamine in ethanol (15\u2005ml). The reaction mixture was stirred for 5\u2005h under reflux. Single crystals of the title compound suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution .Uiso(H) = 1.5Ueq(O). Other H atoms were fixed geometrically and treated as riding with C\u2014H = 0.96\u2005\u00c5 (meth\u00adyl) or 0.93\u2005\u00c5 (aromatic), Uiso(H) = 1.2Ueq(C) or 1.5Ueq(Cmeth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989020011652/zl2794sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020011652/zl2794Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020011652/zl2794Isup3.cmlSupporting information file. DOI: 2025323CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "CENP\u2010A\u2010containing nucleosomes. To preserve centromere identity, CENP\u2010A must be escorted to centromeres by a CENP\u2010A\u2010specific chaperone for deposition. Despite this essential requirement, many eukaryotes differ in the composition of players involved in centromere maintenance, highlighting the plasticity of this process. In humans, CENP\u2010A recognition and centromere targeting are achieved by HJURP and the Mis18 complex, respectively. Using X\u2010ray crystallography, we here show how Drosophila CAL1, an evolutionarily distinct CENP\u2010A histone chaperone, binds both CENP\u2010A and the centromere receptor CENP\u2010C without the requirement for the Mis18 complex. While an N\u2010terminal CAL1 fragment wraps around CENP\u2010A/H4 through multiple physical contacts, a C\u2010terminal CAL1 fragment directly binds a CENP\u2010C cupin domain dimer. Although divergent at the primary structure level, CAL1 thus binds CENP\u2010A/H4 using evolutionarily conserved and adaptive structural principles. The CAL1 binding site on CENP\u2010C is strategically positioned near the cupin dimerisation interface, restricting binding to just one CAL1 molecule per CENP\u2010C dimer. Overall, by demonstrating how CAL1 binds CENP\u2010A/H4 and CENP\u2010C, we provide key insights into the minimalistic principles underlying centromere maintenance.Centromeres are microtubule attachment sites on chromosomes defined by the enrichment of histone variant CAL1 binds both pre\u2010nucleosomal CENP\u2010A/H4 histone dimers and the centromere receptor CENP\u2010C, thus combining the activities of mammalian counterparts HJURP and Mis18 complex.X\u2010ray crystallography reveals how the evolutionary distinct histone chaperone Centromeres are specialised chromosomal regions that act as a platform for the assembly of kinetochores, the microtubule anchoring sites essential for chromosome segregation during mitosis and meiosis , but lack clear homologues of HJURP and the subunits of the Mis18 complex. Instead, fly\u2010specific CAL1 appears to combine the roles of both HJURP and the Mis18 complex: pre\u2010nucleosomal CENP\u2010A recognition and its targeting to the centromere for deposition, respectively is sufficient to interact with CAL1 and H4. Subsequently, using CAL11\u2013160, CENP\u2010A144\u2013255 and H4, we reconstituted a truncated protein complex (His\u2010CAL11\u2013160\u2013CENP\u2010A144\u2013225\u2013H4). The molecular weights (MW) measured for His\u2010CAL11\u2013160\u2013CENP\u2010A101\u2013225\u2013H4 and His\u2010CAL11\u2013160\u2013CENP\u2010A144\u2013225\u2013H4 using size\u2010exclusion chromatography combined multi\u2010angle light scattering (SEC\u2010MALS) are 47.0\u00a0\u00b1\u00a00.9 and 43.4\u00a0\u00b1\u00a00.8\u00a0kDa, respectively residues 1\u2013200 predicted to fold into \u03b1 helices Fig\u00a0A and B. vely Fig\u00a0C. These 1\u2013160\u2013CENP\u2010A101\u2013225\u2013H4 and CAL11\u2013160\u2013CENP\u2010A144\u2013225\u2013H4 yielded two different crystal forms: form I that diffracted X\u2010rays to about 3.5\u00a0\u00c5 and form II that diffracted anisotropically to about 4.4\u00a0\u00c5 (Table\u00a0Drosophila melanogaster (dm) H3/H4 heterodimer . Purified recombinant CAL11\u2013160\u2013CENP\u2010A101\u2013225\u2013H4 complex was cross\u2010linked using EDC (solid lines), a zero\u2010length cross\u2010linker that covalently links carboxylate groups of Asp or Glu residues with primary amines of Lys and N\u2010terminus, or hydroxyl group of Ser, Thr and Tyr, or BS3 (dashed lines), a cross\u2010linker that covalently links amine to amine or hydroxyl group of Ser, Thr and Tyr. The cross\u2010linked peptides were analysed by mass spectrometry to identify intra\u2010 and intermolecular contacts is very similar to human CENP\u2010A/H4 (PDB: 3NQJ) and hydrophobic (involving CAL1\u00a0L11 and M14) interactions, the C\u2010terminal half, mainly aa W22 and F29, is sandwiched between CENP\u2010A \u03b12 and H4 \u03b13 and without \u03b1 helical elements (CAL11\u201350) to bind CENP\u2010A/H4 did not show a noticeable difference under the conditions (buffer containing at least 1M NaCl) needed for CENP\u2010A/H4 solubility. We speculate that in a cellular context post\u2010translational regulation such as phosphorylation or/and other intermolecular interaction involving the downstream helical segments of CAL1 might modulate CENP\u2010A/H4 binding dynamics required for correct CENP\u2010A recruitment at centromeres. This may be a possible explanation for why CAL11\u201350 is not sufficient for CENP\u2010A recruitment in cells and tested their ability to interact with His\u2010CAL11\u2013160 in a nickel\u2010NTA pull\u2010down assay and a four\u2010stranded (\u03b24\u2010\u03b29\u2010\u03b26\u2010\u03b27) and analysed their oligomeric structure by measuring the MW using SEC\u2010MALS , the overall fold conferring the \u03b2 barrel structure is conserved. However, two loop regions show striking conformational variation as compared with their equivalent regions in budding yeast CENP\u2010C, Mif2p . The structure reported here and PDB: 6O2K are nearly identical and superpose well with an RMSD of 0.27\u00a0\u00c5 . Next, samples comprising a SEC purified untagged complex were analysed. Both the mass and sedimentation coefficient are consistent with a 2:1 complex, but not with a 2:2 complex . Thus, the AUC together with the crystal structure shows that CENP\u2010C cupin dimer binds just one copy of CAL1 at any given time.Although CAL1 binding by CENP\u2010C involves just a cupin monomer, only one of the two cupin monomers was observed to interact with CAL1, while the equivalent CAL1 binding site of the dimeric counterpart was empty in the crystal structure. We speculate that the other binding site might be sterically hindered by the remaining residues of CAL1 not seen in the crystal structure, thus not allowing a second monomer of CAL1 to bind. This agrees with our previous observation that CAL1AUC) Fig\u00a0C demonst101\u2013225 and H4\u00a0in a similar manner to CAL11\u2013160. The resulting folded complex was analysed by SEC on its own and mixed with 1.2 molar excess of\u00a0His\u2010CENP\u2010C1264\u20131411 compared to His\u2010CAL11\u2013160\u2010LL\u2010841\u2013979/CENP\u2010A101\u2013225/H4 (14.62\u00a0ml) and His\u2010CENP\u2010C1264\u20131411 (16.05\u00a0ml) on their own results in genome instability, often leading to cell death. To maintain centromere identity defined by the enrichment of CENP\u2010A containing nucleosome, the CENP\u2010A\u2010specific chaperone (HJURP in humans and Scm3 in yeast) escorts CENP\u2010A until its incorporation into the centromeric chromatin and Schizosaccharomyces pombe (Cnp3) CENP\u2010C cupin domains, with striking differences in the mode of dimerisation and used directly in ligation\u2010independent cloning (LIC) into bacterial expression vectors. Smaller fragments were amplified using PCR and then used for LIC. CAL11\u2013160\u2010LL\u2010841\u2013979 was produced using homologous PCR.Codon optimised All mammalian expression vectors used in this study were constructed in a pN2\u2010CMV vector.101\u2013225 was generated in Gold (Agilent) grown in 2XTY media. His\u2010CENP\u2010C1264\u20131411 was expressed in Rosetta (DE3) (Novagen) cells, using 2XTY. His\u2010SUMO\u2010CAL1841\u2013979 was expressed in BL21 (DE3) Gold in LB. After reaching O.D ~\u00a00.6 at 37\u00b0C, cultures were induced at 18\u00b0C using 0.3\u00a0mM IPTG before being purified under native and denaturing conditions. Under native conditions for all protein, pellets were resuspended in a buffer containing 20\u00a0mM Tris\u2013HCl pH 8.0, 100\u00a0mM NaCl, 35\u00a0mM imidazole and 2\u00a0mM \u03b2ME and supplemented with 1\u00a0mM PMSF and cOmplete EDTA\u2010free (Sigma) before lysing by sonication. Clarified lysates were applied onto a HisTrap\u00ae HP column. For His\u2010CAL11\u2013160, His\u2010CAL11\u201350 and His\u2010CAL11\u2013160\u2010LL\u2010841\u2013979, HisTrap\u00ae HP columns were then washed with 60 CV of lysis buffer, 20 CV of 20\u00a0mM Tris\u2013HCl pH 8.0, 1\u00a0M NaCl, 35\u00a0mM imidazole, 50\u00a0mM KCl, 10\u00a0mM MgCl2, 2\u00a0mM ATP and 2\u00a0mM \u03b2ME and then 10 CV of lysis buffer. Proteins were then eluted in 20\u00a0mM Tris\u2013HCl pH 8.0, 100\u00a0mM NaCl, 500\u00a0mM imidazole and 2\u00a0mM \u03b2ME. For His\u2010SUMO\u2010CAL1841\u2013979 and His\u2010CENP\u2010C1264\u20131411, HisTrap\u00ae HP columns were washed with 80 CV of lysis buffer. Protein was eluted using 20\u00a0mM Tris\u2013HCl pH 8.0, 100\u00a0mM NaCl, 500\u00a0mM imidazole and 2\u00a0mM \u03b2ME, and fractions containing protein were dialysed overnight against 20\u00a0mM Tris\u2013HCl pH 8.0, 500\u00a0mM NaCl and 2\u00a0mM DTT for His\u2010CENP\u2010C1264\u20131411 and 20\u00a0mM Tris\u2013HCl pH 8.0, 100\u00a0mM NaCl and 2\u00a0mM DTT for His\u2010SUMO\u2010CAL1841\u2013979. His\u2010SUMO\u2010CAL1841\u2013979 was applied to a HiTrap\u00ae Q HP column and eluted with a gradient of 20\u00a0mM Tris\u2013HCl (pH 8.0), 1\u00a0M NaCl and 2\u00a0mM DTT.His\u2010CAL1Tags were removed by incubation with 3C or TEV overnight followed by a reverse affinity step to remove His\u2010SUMO. Proteins were purified by SEC using either a Superdex 200 increase 10/300 GL, Superdex 75 10/300 GL or Superdex 75 increase 10/300 GL column .1\u2013160, His\u2010CAL11\u201350 and His\u2010CAL11\u2013160\u2010LL\u2010841\u2013979, pellets were suspended in 2\u00a0ml/g of wet pellet of 20\u00a0mM Tris\u2013HCl pH 8.0, 500\u00a0mM NaCl, 25\u00a0mM imidazole, 7\u00a0M urea and 2\u00a0mM \u03b2ME, and incubated for 1\u00a0h at 4\u00b0C with rotation. DNA was sheared by sonication before clarifying by centrifugation. Lysate was then incubated with 10\u00a0ml of HisPur\u2122 Ni\u2010NTA resin (Thermo Fisher Scientific) overnight, before washing with 60 CV of buffer, 20 CV of 20\u00a0mM Tris\u2013HCl (pH 8.0), 1\u00a0M NaCl, 25\u00a0mM imidazole, 7\u00a0M urea and 2\u00a0mM \u03b2ME, 10 CV of 500\u00a0mM NaCl buffer before eluting with 20\u00a0mM Tris\u2013HCl (pH 8.0), 500\u00a0mM NaCl, 500\u00a0mM imidazole, 7\u00a0M guanidine HCl and 2\u00a0mM \u03b2ME.For denatured His\u2010CAL1To refold histones with and without CAL1, histones were resuspended in 20\u00a0mM Tris\u2013HCl pH 7.5, 7\u00a0M guanidine HCl and 2\u00a0mM \u03b2ME and mixed with equimolar amounts of proteins needed. Proteins were then dialysed for 2\u00a0h at 4\u00b0C against 200\u00a0ml of 20\u00a0mM Tris\u2013HCl pH 7.5, 7\u00a0M guanidine HCl and 2\u00a0mM \u03b2ME; then, 2\u00a0l of 10\u00a0mM Tris\u2013HCl pH 7.5, 2\u00a0M NaCl, 1\u00a0mM EDTA and 5\u00a0mM \u03b2ME was slowly added overnight using a peristaltic pump. If needed, refolded protein was further dialysed against a lower salt concentration solvent; if not, complexes were purified by SEC using either a Superdex 200 increase 10/300 GL or Superdex 75 increase 10/300 GL column .1\u2013160\u2010CENP\u2010A101\u2013225\u2010H4, 27\u00a0mg/ml of complex in a buffer of 20\u00a0mM Tris\u2013HCl pH 8.0, 1\u00a0M NaCl and 5\u00a0mM DTT crystallised after about a year in 0.01\u00a0M Cobalt (II) chloride hexahydrate, 0.1\u00a0M MES pH 6.5 and 1.8\u00a0M ammonium sulphate at 18\u00b0C. For CAL11\u2013160\u2010CENP\u2010A144\u2013225\u2010H4, protein in 20\u00a0mM Tris\u2013HCl pH 8.0, 1\u00a0M NaCl and 2\u00a0mM DTT was concentrated to 17\u00a0mg/ml and crystallised in C11, 0.2\u00a0M lithium sulphate, 0.1\u00a0M Tris pH 8.5 and 30% PEG 4000. An optimisation screen was set up in a 24\u2010well format, using half the original concentrations of protein. Tris\u2013HCl pH 8.5 was kept at 0.1\u00a0M, while concentrations of lithium sulphate varied from 0.1 to 0.3\u00a0M and PEG 4000 varied from 24 to 34%.Crystallisation trials were performed using a nanolitre Crystal Gryphon robot (Art Robbins) and grown by vapour diffusion methods. For CAL11264\u20131411 was screened at 15\u00a0mg/ml in 20\u00a0mM Tris\u2013HCl pH 8.0, 500\u00a0mM NaCl and 2\u00a0mM DTT against several commercial and homemade screens at 18\u00b0C. Crystals were obtained in around 13% of all conditions tested. His\u2010CENP\u2010C1264\u20131411\u2010CAL1841\u2013979 complex was made in 20\u00a0mM Tris\u2013HCl pH 8.0, 100\u00a0mM NaCl and 2\u00a0mM DTT and used with Structure 1\u00a0+\u00a02 and JCSG+ (Molecular Dimensions) at 15\u00a0mg/ml at 4\u00b0C. Crystals were briefly transferred to a cryoprotectant solution (either oil or the mother liquor supplemented with 40% peg 3350) before directly flash cooled in liquid nitrogen and analysed on beamlines i03 and i04\u20101 at the Diamond Light Source .Cleaved CENP\u2010C1\u2013160\u2013CENP\u2010A/H4 form I, CENP\u2010C1264\u20131411; CENP\u2010C1264\u20131411\u2010CAL1841\u2013979) and i04\u20101 (CAL11\u2013160\u2013CENP\u2010A/H4 form II), at the Diamond Light Source . Data were processed using the software pipeline available at Diamond Light Source that relies on XDS, CCP4, CCTBX, AutoPROC and STARANISO , CENP\u2010C1264\u20131411 and CENP\u2010C1264\u20131411\u2010CAL1841\u2013979 structures were determined by molecular replacement with the program PHASER . Data collection, phasing and refinement statistics are shown in Table\u00a0Diffraction data were collected on beamlines i03 run at 180\u00a0V for 1\u00a0h in MES buffer. Gels were then stained with Coomassie Blue, and scanned gel images were analysed and quantified with ImageJ.Ni\u2010NTA pull\u2010down assays were performed using His\u2010CAL1Size\u2010exclusion chromatography coupled to UV, static light scattering and refractive index detection was used to determine the molecular mass of proteins and protein complexes in solution. Injections of 100\u00a0\u03bcl of 1\u20135\u00a0mg/ml material were used.1\u2013160\u2010CENP\u2010A101\u2013225\u2010H4 (\u2202A280\u00a0nm/\u2202c\u00a0=\u00a00.67 AU.ml/mg), His\u2010CAL11\u2013160\u2010CENP\u2010A144\u2013225\u2010H4 (\u2202A280\u00a0nm/\u2202c\u00a0= 0.75 AU.ml/mg) and His\u2010CAL11\u201350\u2010CENP\u2010A101\u2013225\u2010H4 (\u2202A280\u00a0nm/\u2202c\u00a0= 0.55 AU.ml/mg) were run on a Superdex 200 increase 10/300 GL size\u2010exclusion column pre\u2010equilibrated in 50\u00a0mM HEPES pH 8.0, 2\u00a0M NaCl and 1\u00a0mM TCEP at 22\u00b0C with a flow rate of 0.5\u00a0ml/min. His\u2010CENP\u2010C1264\u20131411 L1357E/M1407E (\u2202A280\u00a0nm/\u2202c\u00a0=\u00a00.75\u00a0AU.ml/mg) was run on a Superdex 200 increase 10/300 GL size\u2010exclusion column pre\u2010equilibrated in 50\u00a0mM HEPES pH 8.0, 300\u00a0mM NaCl and 1\u00a0mM TCEP at 22\u00b0C with a flow rate of 0.5\u00a0ml/min. CENP\u2010C1264\u20131411 (\u2202A280\u00a0nm/\u2202c\u00a0=\u00a00.84\u00a0AU.ml/mg) was run at 4\u00b0C on a Superdex 75 increase 10/300 GL size\u2010exclusion column pre\u2010equilibrated in 50\u00a0mM HEPES pH 8.0, 100\u00a0mM NaCl and 1\u00a0mM TCEP.For His\u2010CAL1280\u00a0nm were analysed by a homo\u2010polymer model using the parameters stated for each protein, \u2202n/\u2202c\u00a0=\u00a00.185\u00a0ml/g and buffer RI value of 1.335. The mean standard error in the mass accuracy determined for a range of protein\u2013protein complexes spanning the mass range of 6\u2013600\u00a0kDa is \u00b1\u00a01.9%.Light scattering, refractive index (RI) and A3 was used to cross\u2010link 10\u00a0\u03bcg of protein for 2\u00a0h at RT. The reactions were quenched with final concentration 100\u00a0mM Tris\u2013HCl or 5\u00a0mM ammonium bicarbonate, respectively, before separation on Bolt\u2122 4\u201312% Bis\u2010Tris Plus gels (Invitrogen). Following previously established protocol and 66\u00a0\u03bcg sulpho\u2010NHS (Thermo Fisher Scientific) were used to cross\u2010link 10\u00a0\u03bcg of protein for 1.5\u00a0h at RT. 30\u00a0\u03bcg of BSet\u00a0al (5 cells per well. Cells were transfected using X\u2010tremeGENE DNA Transfection Reagent (Roche) according to the manufacturer's instructions, using 200\u00a0ng of plasmid DNA. Transfected cells were analysed by immunofluorescence 72\u00a0h post\u2010transfection.Schneider S2 cells containing the LacO array (L2\u20104_LacO_LexA_Clone11) were generated as described in Mendiburo et\u00a0al . Schneidet\u00a0al, 2 incubator. Cells were seeded in 10\u00a0cm dishes, a day prior to transfection at a density of 2.5\u00a0\u00d7\u00a0106 cells per well. Transfections were performed with Lipofectamine 3000 (Life Technologies) according to the manufacturer's instructions, using 15\u00a0\u03bcg of plasmid DNA and Opti\u2010MEM I reduced serum medium (Life Technologies). Next day, cells were washed once with 1\u00d7DPBS, trypsinised, counted and re\u2010plated on poly\u2010lysine\u2010coated coverslips in 6\u2010well plates at a density of 106 cells per\u00a0well. Downstream experiments were performed 3\u00a0days post\u2010transfection.U2OS cells containing 200 copies of an array of 256 tandem repeats of the 17\u00a0bp LacO sequence on chromosome 1 overnight at 4\u00b0C in a humidified chamber and were used in 1:100 dilution unless otherwise stated: myc (Abcam\u2010ab9106), V5 (Invitrogen\u2010R96025) and HA . Secondary antibodies coupled to Alexa Fluor 555 and 647 (Invitrogen) were used at 1:100 dilutions. Counterstaining of DNA was performed with DAPI (5\u00a0\u03bcg/ml), and coverslips were mounted on the slides with 30\u00a0\u03bcl of SlowFade\u00ae Gold antifade reagent.Cells were washed once in PBS and then fixed with 3.7% formaldehyde in 0.1% Triton X\u2010100 in 1\u00d7 PBS (PBST) for 8\u00a0min at RT. Following fixation, the slides were washed once in PBST and then blocked in Image\u2010iTz\u2010stacks of 0.2\u00a0\u03bcm increments, using a 100\u00d7 oil immersion objective on a DeltaVision RT Elite Microscope and a CoolSNAP HQ Monochrome camera. All images were deconvolved using the aggressive deconvolution mode on a SoftWorx Explorer Suite (Applied Precision) and are shown as quick projections of maximum intensity. For U2OS cells, the mean fluorescence intensity of the protein of interest was measured at the LacO spot, and then the mean fluorescence intensity in the nucleus (background) was subtracted from this value. For S2 cells, the mean fluorescence intensity of the protein of interest was measured at the LacO spot, and then the mean fluorescence intensity of three spots around the LacO spot was subtracted from this value. 25\u201350 cells were analysed per biological replicate, and a minimum of three independent biological replicates were quantified per experiment.All IF images were taken as 50 et\u00a0al, et\u00a0al, Sedimentation velocity and SE experiments were performed using a Beckman Coulter XL\u2010I analytical ultracentrifuge equipped with an An\u201050 Ti eight\u2010hole rotor. Depending on their concentration, samples were loaded into 12 (low concentration) or 3\u00a0mm (high concentration) pathlength charcoal\u2010filled epon double\u2010sector centrepieces, sandwiched between two sapphire windows. For SV, samples were equilibrated at 4\u00b0C in vacuum for 6\u00a0h before running at 49\u00a0k rpm. For SE, data were recorded at 26\u00a0k rpm. The laser delay, brightness and contrast were pre\u2010adjusted at 3\u00a0k rpm to acquire the best quality interference fringes. Data were collected using Rayleigh interference and absorbance optics recording radial intensity or absorbance at 280\u00a0nm. For SV, data were recorded between radial positions of 5.65 and 7.25\u00a0cm, with a radial resolution of 0.005\u00a0cm and a time interval of 7\u00a0min, and analysed with the program SEDFIT Schuck, using a t\u2010test or Mann\u2013Whitney test. For each statistical test, P\u2010value of <\u00a00.05 was considered significant.Data are representative of at least three independent experiments, unless otherwise stated. Statistics were performed using GraphPad Prism, version 7.0e , using an unpaired two\u2010tailed AAJ and PH conceived the project. BM\u2010P, VL, PH and AAJ designed experiments. BM\u2010P,VL, OB, MAA and JZ performed experiments. JR provided expertise and feedback. BM\u2010P and AAJ wrote the manuscript with input from all authors.The authors declare that they have no conflict of interest.AppendixClick here for additional data file.Expanded View Figures PDFClick here for additional data file.Review Process FileClick here for additional data file."} +{"text": "In the crystal structure of the title compound inter\u00admolecular hydrogen-bonding inter\u00adactions and weak C\u2014H\u22ef\u03c0 inter\u00adactions between the constituents lead to the formation of a three-dimensional network. Hirshfeld surface analysis revealed that H\u22efH inter\u00adactions dominate the crystal packing. 22H17NO2\u00b7C3H7NO, was synthesized by condensation of an aromatic aldehyde with a secondary amine and subsequent reduction. It was crystallized from a di\u00admethyl\u00adformamide solution as a monosolvate, C22H17NO2\u00b7C3H7NO. The aromatic mol\u00adecule is non-planar with a dihedral angle between the mean planes of the aniline moiety and the methyl anthracene moiety of 81.36\u2005(8)\u00b0. The torsion angle of the Car\u00adyl\u2014CH2\u2014NH\u2014Car\u00adyl backbone is 175.9\u2005(2)\u00b0. The crystal structure exhibits a three-dimensional supra\u00admolecular network, resulting from hydrogen-bonding inter\u00adactions between the carb\u00adoxy\u00adlic OH group and the solvent O atom as well as between the amine functionality and the O atom of the carb\u00adoxy\u00adlic group and additional C\u2014H\u22ef\u03c0 inter\u00adactions. Hirshfeld surface analysis was performed to qu\u00adantify the inter\u00admolecular inter\u00adactions.The title compound, C Schiff bases belong to a class of organic compounds that are formed by the condensation reaction of a carbonyl carbon with an aliphatic/aromatic amine, resulting in the formation of a characteristic imine bond (\u2013HC=N\u2013). Many Schiff bases exhibit activities of biological and pharmaceutical significance. Moreover, Schiff bases are actively used as organic linkers for building metal complexes with inter\u00adesting properties.et al., 2017et al., 2014et al., 2012), anti\u00admicrobial . The title compound crystallizes with a di\u00admethyl\u00adformamide (DMF) solvent mol\u00adecule in a 1:1: ratio. Both anthraldehyde and PABA have shown anti\u00adcancer (Pavitha ar\u00adyl\u2014CH2\u2014NH\u2014Car\u00adyl backbone (C9\u2014C8\u2014N1\u2014C5) is 175.9\u2005(2)\u00b0. The C8\u2014N1 bond length of 1.452\u2005(3)\u2005\u00c5 is in agreement with the corresponding bond length of 1.457\u2005(3)\u2005\u00c5 in the solvent-free compound \u00adisophthalic acid containing gadolinium = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020005393/wm5548sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020005393/wm5548Isup2.hklStructure factors: contains datablock(s) I. DOI: 1982147CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-heterocyclic carbene precursor, zwitterionic 3,3\u2032-bis\u00ad(propane-1-sulfonate) crystallized as its dihydrate and was characterized by single-crystal X-ray diffraction, revealing point group symmetry A known 16H26N4O6S2\u00b72H2O, a water-soluble di-N-heterocyclic carbene ligand precursor was determined using a single crystal grown by the slow cooling of a hot N,N-di\u00admethyl\u00adformamide solution of the compound. The dihydrate crystallizes in the monoclinic space group P21/c, with half of the zwitterionic mol\u00adecule and one water mol\u00adecule of crystallization in the asymmetric unit. The remaining part of the mol\u00adecule is completed by inversion symmetry. In the mol\u00adecule, the imidazole ring planes are parallel with a plane-to-plane distance of 2.741\u2005(2)\u2005\u00c5. The supra\u00admolecular network is consolidated by hydrogen bonds of medium strength between the zwitterionic mol\u00adecules and the water mol\u00adecules of crystallization, as well as by \u03c0\u2013\u03c0 stacking inter\u00adactions between the imidazole rings of neighbouring mol\u00adecules and C\u2014H\u22efO hydrogen-bonding inter\u00adactions.The crystal structure of the title compound, C To the best of our knowledge, this is the first crystal structure determination of an alkyl\u00adene-bridged di-imidazolium salt with \u03c9-propyl\u00adsulfonate wingtips. Compound 1 is known from the literature bis\u00ad. No molecules of the solvent, DMF, from which the crystals were obtained, are built into the lattice.The di-1 \u2005\u00c5, S1\u2014O1 = 1.446\u2005(2)\u2005\u00c5, S1\u2014O2 = 1.450\u2005(2)\u2005\u00c5, S1\u2014O3 = 1.453\u2005(2)\u2005\u00c5, and angles O1\u2014S1\u2014O2 = 112.16\u2005(14)\u00b0, O1\u2014S1\u2014O3 = 111.80\u2005(14)\u00b0, and O2\u2014S1\u2014O3 = 112.80\u2005(15)\u00b0. As a result of the point group symmetry The zwitterionic mol\u00adecule of 1 Fig.\u00a02 is compo\u2005\u00c5 Fig.\u00a03 from eacx, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z; centroid-to-centroid distance of 3.9541\u2005(17)\u2005\u00c5, slippage 1.622\u2005\u00c5, Fig.\u00a05The water mol\u00adecules bridge adjacent zwitterionic mol\u00adecules through hydrogen bonds of medium strength with sulfonate O atoms as acceptor groups into ribbons aligned parallel to [001] Table\u00a01. \u03c0\u2013\u03c0 staet al., 2016N-heterocyclic carbene ligand precursor mol\u00adecules where two N-\u03c9-sulfonato\u00adpropyl-imidazolium units are connected through an \u03b1,\u03c9-alkyl\u00adene bridge. Crystals of similar sulfoalkyl-imidazolium di-NHC precursors containing aromatic linkers have been grown by Kohmoto et al. were also reported.A search of the Cambridge Structural Database ], refcode: YAVROF, and [Pd(2)2], refcode: YAVXAX, crystallize in space group type PP21/c.The Pd complexes [PdCl1 was synthesized according to the method of Papini et al. . Analytical data: 13C{1H}-NMR \u03b4 [ppm], 135.6, 122.5, 48.8, 47.8, 47.2, 26.2, 25.0; ESI\u2013MS , m/z observed 435.1359, calculated value for C16H27N4O6S2, ([M - H]+): 435.1367). For recrystallization, 1 was suspended in DMF and heated to approximately 373\u2005K, then filtered and left overnight to slowly cool down to room temperature. Single crystals, suitable for X-ray analysis, were obtained as colourless prisms after storing the solution in open glass vials in a refrigerator at 278\u2005K. A possible source of water is the employed DMF, which is hygroscopic and easily adsorbs water from a humid atmos\u00adphere. The same type of prismatic crystals were also grown from hot water, revealing a very similar unit cell. However, these crystals were of poor quality, and the best Rint value was very high, 0.19.Compound al. 2009. 4.4\u2005mmoUiso(H) = 1.5Ueq(O).Crystal data, and details of data collection and structure refinement are summarized in Table\u00a0210.1107/S2056989020009779/wm5571sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020009779/wm5571Isup3.cdxSupporting information file. DOI: 10.1107/S2056989020009779/wm5571Isup4.hklStructure factors: contains datablock(s) I. DOI: 2017141CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Nature Communications 10.1038/s41467-020-14509-4, published online 31 January 2020.Correction to: The original version of this Article contained an error in the information provided in the legend for Supplementary Fig. 4D. The text should read: 1\u2009=\u2009UA-GalNAc4S; 2\u2009=\u2009UA-GalNAc4S incubated with B.theta PBS washed cells for 1h; 3\u2009=\u2009UA-GalNAc4S incubated with B.theta PBS washed cell for 24h; 4\u2009=\u2009UA-GalNAc4S incubated with sonicated B.theta PBS washed cells for 1h; 5\u2009=\u2009UA-GalNAc4S incubated with sonicated B.theta PBS washed cell for 24h; 6\u2009=\u2009GalNAc; 7\u2009=\u2009GalNAc6S; 8\u2009=\u2009GalNAc6S incubated with B.theta PBS washed cells for 1h; 9\u2009=\u2009GalNAc6S incubated with B.theta PBS washed cells for 24h; 10\u2009=\u2009GalNAc6S incubated with sonicated B.theta PBS washed cell for 1h and 11\u2009=\u2009GalNAc6S incubated with sonicated B.theta PBS washed cells for 24.This has been corrected in both the PDF and HTML versions of the Article."} +{"text": "II\u2010mediated base pair than a rigid HgII\u2010mediated base pair within the base stack of a double helix.Oligonucleotides incorporating a central C\u2010nucleoside with either a rigid or flexible benzaldoxime base moiety have been synthesized, and the hybridization properties of their metallacyclic derivatives have been studied by UV melting experiments. In all cases, the metallated duplexes were less stable than their unmetallated counterparts, and the metallacyclic nucleobases did not show a clear preference for any of the canonical nucleobases as a base\u2010pairing partner. With palladated oligonucleotides, increased flexibility translated to less severe destabilization, whereas the opposite was true for the mercurated oligonucleotides; this reflects the greater difficulties in accommodating a rigid Pd Take the strain: A flexible PdII\u2010mediated base pair of a benzaldoxime palladacycle disrupts oligonucleotide hybridization less than its rigid analogue, whereas the opposite is true for HgII\u2010mediated base pairs. With a base pair of an incompatible geometry, increased flexibility alleviates some of the strain; with a strain\u2010free base pair it merely leads to an increased entropic penalty of hybridization. II for the canonical nucleobasesDouble\u2010helical oligonucleotides incorporating metal\u2010mediated base pairsII\u2010mediated base pairs have been reported since 1999,II\u2010mediated base pairing remained elusive for a long time and even today the most compelling results have been obtained on duplexes having the PdII\u2010mediated base pair(s) at a terminal position.II\u2010mediated base pair could stem from suboptimal geometries of the base pairs reported so far. Indeed, a thorough study of PdII\u2010mediated base pairing between terpyridine and various azoles has revealed that a planar base pair was only formed with 1\u2010methyltetrazole, having no substituents (not even hydrogens) facing the PdII\u2010terpyridine complex.II center somewhat more exposed than the PdII\u2010terpyridine complex but achieving a coplanar orientation of the palladacycle and the canonical nucleobase may still be difficult. As the steric requirements for base pairing are less strict at the termini than within the base stack, the stabilization provided by the high binding affinity of PdII is not offset by strain imposed by the unfavorable geometry of the PdII\u2010mediated base pair.Although a number of PdII ion in the same position but the former \u20103\u2010[(tert\u2010butyldimethylsilyl)oxy]\u20102,3\u2010dihydrofuran\u20102\u2010yl}methanol and 2\u2010(4\u2010bromophenyl)\u20101,3\u2010dioxane. Compound 1 was then desilylated to the ketone intermediate 2 which, in turn, was reduced to the protected C\u2010nucleoside 3. 5\u2032\u2010Dimethoxytritylation of compound 3 afforded intermediate 4 and subsequent 3\u2032\u2010phosphitylation the phosphoramidite building block 5. Synthesis of compound 5 has been described previouslySynthesis of the protected benzaldehyde C\u2010nucleoside 8 and its phosphoramidite building block 10 were synthesized following the pathway outlined in Scheme\u20057 was first prepared by a Mitsunobu reaction between 3,5\u2010di\u2010O\u2010benzyl\u2010C\u2010hydroxymethyl\u20102\u2010deoxy\u2010\u03b2\u2010d\u2010ribofuranose (6)N\u2010hydroxhphthalimide. Removal of the benzyl protections by Pd(OH)2/C\u2010catalyzed hydrogenation afforded compound 8. Preparation of compound 8 has been reported previously8 was 5\u2032\u2010dimethoxytritylated to give compound 9 which, in turn, was 3\u2032\u2010phosphitylated to give the phosphoramidite building block 10. These last two steps were carried out as described in the literature.Phthaloyl\u2010protected aminooxymethyl C\u2010nucleoside ON1x, ON1y and ON1b were assembled on an automated DNA/RNA synthesizer by the conventional phosphoramidite strategy, employing an extended coupling time for building blocks 5 and 10. Normal (ca. 99\u2009%) coupling yields were observed throughout the synthesis. The acetal and phthaloyl protections of the benzaldehyde and aminooxymethyl residues were removed on\u2010support by treatment with dichloroacetic acid in wet CH2Cl2 or hydrazine acetate in pyridine, respectively. The newly exposed aminooxy function was immediately allowed to react with either 3\u2010hydroxybenzaldehyde or benzaldehyde. Finally, oligonucleotides ON1x, ON1y and ON1b were released from the solid support and deprotected by conventional ammonolysis and purified by RP\u2010HPLC. ON1x eluted as two barely separable peaks, presumably attributable to E and Z isomers of the oxime bond. The stereoselectivity of oximation of benzaldehydes is known to be highly sensitive to the reaction conditionsortho\u2010mercuration), the oligonucleotide environment probably affects as well. Unfortunately, the isolated amounts of the putative isomers of ON1x were not sufficient for more detailed characterization.Table\u2005ON1b was treated with an aqueous solution of methoxylamine to convert the central benzaldehyde residue into an O\u2010methylbenzaldoxime residue. Oligonucleotide ON1z thus obtained was purified by RP\u2010HPLC and cyclopalladated in an aqueous solution of Li2PdCl4. The product mixture was fractioned on RP\u2010HPLC to afford the palladacyclic oligonucleotide ON1z\u2212Pd. Finally, oligonucleotides ON1b, ON1x, ON1y, ON1z and ON1z\u2212Pd were characterized by ESI\u2010TOF\u2010MS and quantified by UV spectrophotometry. Synthesis of the modified oligonucleotides ON1x\u2212Hg and ON1y\u2212Pd has been described previously.ON1x, ON1x\u2212Hg, ON1y, ON1y\u2212Pd, ON1z and ON1z\u2212Pd were studied by conventional UV melting experiments. Each modified oligonucleotide was mixed with each of the unmodified counterparts ON2a, ON2c, ON2g and ON2t, pairing the artificial residue with adenine, cytosine, guanine or thymine, respectively. The oligonucleotide concentrations of the samples were 1.0\u2005\u03bcM, the pH\u20057.4 (20\u2005mM cacodylate buffer) and the ionic strength 0.10\u2005M (adjusted with sodium perchlorate). Melting profiles for unmodified duplexes of otherwise identical sequence but with cytosine in place of the modified residue have been reported previously.Hybridization properties of the modified oligonucleotides ON1x\u2009\u22c5\u2009ON2t, ON1x\u2212Hg\u2009\u22c5\u2009ON2t, ON1y\u2009\u22c5\u2009ON2t, ON1y\u2212Pd\u2009\u22c5\u2009ON2t, ON1z\u2009\u22c5\u2009ON2t and ON1z\u2212Pd\u2009\u22c5\u2009ON2t as representative examples . With the notable exception of ON1z\u2212Pd, monophasic sigmoidal melting curves were observed in most cases. Melting temperatures of the unmetallated duplexes formed by ON1y, ON1z and the faster\u2010eluting isomer of ON1x ranged from 33 to 36\u2009\u00b0C, typical for 11\u2010mer oligodeoxynucleotides containing a single mismatch . Apparently PdII\u2010mediated base pairing by the benzaldoxime palladacycles of ON1y\u2212Pd and ON1z\u2212Pd causes strain that in the former case is to some extent relieved by the flexible structure of the palladacycle but in the latter case prevents formation of a double helix. Disruption of Watson\u2010Crick base pairing by a neighboring metal\u2010mediated base pairing has been described previously in detail with Ag(I) as the bridging metal ion.Figure\u2005ON1x remained elusive as no consistent pattern of different enthalpies and entropies emerged. The relatively low stability of the duplexes formed by the mercurated oligonucleotide ON1x\u2212Hg compared to respective duplexes incorporating a 5\u2010mercuricytosineON1x\u2212Hg (500\u2013600\u2005J\u2009mol\u22121\u2009K\u22121) than for its unmercurated counterpart ON1x (700\u2013800\u2005J\u2009mol\u22121\u2009K\u22121), it was still considerably higher than the respective value reported for those more stable mercurated duplexes (200\u2013400\u2005J\u2009mol\u22121\u2009K\u22121). HgII\u2010mediated base pairing generally leads to a reduced entropic penalty of hybridization owing to dehydration of the bridging HgII ionON1x\u2212Hg some of this effect is offset by restricted rotation of several \u03c3 bonds. The enthalpies of hybridization were similar for the duplexes formed by ON1x\u2212Hg and those containing 5\u2010mercuricytosine, approximately 200\u2005kJ\u2009mol\u22121.When applicable, thermodynamic parameters of hybridization were determined as described previouslyII\u2010mediated base pairing, detailed thermodynamic data on PdII\u2010mediated base pairing is scanty but reanalysis of UV melting profiles reported previously for octamer duplexes with an unpalladated or palladated benzylamine attached to the 5\u2032 terminus through a flexible linker revealed more negative enthalpies and entropies of hybridization for the palladated oligonucleotides.II\u2010mediated base pairing itself is an enthalpy\u2010driven process, whereas the entropic penalty is consistent with conformational restriction of the flexible linker. In the present case, however, both the enthalpies and the entropies of hybridization were less negative with ON1y\u2212Pd than with ON1y and the differences were actually considerably larger than between ON1x\u2212Hg and ON1x. This result, while in apparent conflict with the data obtained on oligonucleotides featuring terminal palladacyclic modifications, could be understood if formation of the PdII\u2010mediated base prevented formation of some (in the case of ON1y\u2212Pd) or all (in the case of ON1z\u2212Pd) of the canonical Watson\u2010Crick base pairs.Compared to the well\u2010documented HgON1x\u2212Hg and ON1y\u2212Pd were largely similar except for ON1x\u2212Hg\u2009\u22c5\u2009ON2g, in which case the signal at 250\u2005nm was hardly detectable. With ON1z\u2212Pd the spectra were much more distorted and the ellipticity lower, in line with the inability of this oligonucleotide to form a double helix with any of the unmodified counterparts at temperatures above 10\u2009\u00b0C. Gradual thermal diminution of the Cotton effects, consistent with denaturation of the double helix, was observed with all of the duplexes studied.All samples were also analyzed CD spectropolarimetrically for more detailed information on the secondary structures adopted by the oligonucleotides. The spectra were recorded between 10 and 90\u2009\u00b0C at 10\u2009\u00b0C intervals. At the low end of the temperature range, the spectra of the unmetallated duplexes were characteristic of B\u2010type double helices, with negative and positive Cotton effects of nearly equal intensity at 250 and 280\u2005nm, respectively (spectra presented in the Supporting Information). The corresponding spectra of the duplexes formed by II\u2010 or HgII\u2010mediated base pair. The oligonucleotide incorporating a flexible palladacyclic nucleoside formed more stable duplexes than its more rigid counterpart whereas comparison of the present data with those reported previously revealed the opposite to be true for oligonucleotides incorporating organomercury nucleosides. These results can be understood in terms of the different geometries of PdII\u2010 and HgII\u2010mediated base pairs. The former may be unable to assume the coplanar orientation preferred within the base stack of a double helix and in such a case a flexible structure might be able to alleviate some of the resulting strain. The latter, on the other hand, are usually nearly planar and geometrically very similar to canonical Watson\u2010Crick base pairs and in such a case increased flexibility will only result in an increased entropic penalty as the flexible structure is constrained within the base stack.Increased flexibility of the metallated nucleoside analog was found to have profoundly different effects on the melting temperatures of oligonucleotide duplexes containing a central PdGeneral methods: All experiments involving air and/or moisture sensitive compounds were performed using oven\u2010dried glassware under argon atmosphere. For preparation of HPLC elution buffers, freshly distilled triethylamine was used. Other commercially available chemicals were used without further purification unless otherwise stated. The solvents for organic synthesis were of reagent grade and dried over 4\u2005\u00c5 molecular sieves. All reactions were monitored by thin layer chromatography (TLC), performed on Merck 60 (silica gel F254) plates. Chromatographic purification of products was accomplished using flash column chromatography on silica gel (230\u2013400\u2005mesh). 1H, 13C and 31P NMR spectra were recorded in deuterated solvents on a Bruker Biospin 500 or 600\u2005MHz NMR spectrometer. Chemical shifts are quoted relative to the residual solvent peak as internal standard. Mass spectra were recorded on a Bruker Daltonics micrOTOF\u2010Q ESI mass spectrometer.Oligonucleotide synthesis: Synthesis of the modified oligonucleotides ON1x\u2212Hg and ON1y\u2212Pd has been described previously. The other modified oligonucleotides were assembled on an Applied Biosystems 3400 automated DNA/RNA synthesizer by conventional phosphoramidite strategy. For the acetal protected benzaldehyde and the phtahaloyl\u2010protected aminooxymethyl C\u2010nucleoside phosphoramidite building blocks 2 and 8, the coupling time was extended to 300\u2005s. Based on trityl response, all couplings proceeded with normal efficiency. Removal of the acetal protecting group of the benzaldehyde residue was carried out on\u2010support by treatment with 2\u2009% dichloroacetic acid and 1\u2009% H2O in CH2Cl2 at 25\u2009\u00b0C for 2\u2005h.2Cl2 (3.0\u2005mL) or neat benzaldehyde to eventually afford oligonucleotides ON1x and ON1y, respectively. Finally, oligonucleotides ON1x, ON1y and ON1b were released from the solid support and the phosphate and base moieties were deprotected by incubation in 25\u2009% aqueous ammonia at 55\u2009\u00b0C for 12\u2005h and purified by RP\u2010HPLC on a Hypersil ODS C18 column eluting with a linear gradient of MeCN (5\u201330\u2009% over 30\u2005min) in 50\u2005mM aqueous triethylammonium acetate. The flow rate was 1.0\u2005mL\u2009min\u22121 and the detection wavelength 260\u2005nm.ON1b was converted into an O\u2010methylbenzaldoxime residue by treatment with a mixture of methoxylamine hydrochloride (1.0\u2005\u03bcmol) and NaOAc (1.0\u2005\u03bcmol) in H2O (100\u2005\u03bcL) at 25\u2009\u00b0C for 12\u2005h. The crude oligonucleotide ON1z thus obtained was purified by RP\u2010HPLC as described above. For cyclopalladation, ON1z (40\u2005nmol), Li2PdCl4 (60\u2005nmol) and NaOAc (240\u2005nmol) were dissolved in H2O (10\u2005\u03bcL) and the resulting mixture was incubated at 55\u2009\u00b0C for 40\u2005h. The crude product ON1z\u2212Pd was purified by RP\u2010HPLC as described above. As reported previously for other cyclopalladated oligonucleotides,ON1z\u2212Pd eluted as a broad and convoluted peak even after purification, presumably due to the relatively slow ligand\u2010exchange of PdII giving rise to a variety of slow\u2010equilibrating structures involving intra\u2010 as well as interstrand PdII\u2010mediated base pairing. All of the newly synthesized oligonucleotides were characterized by ESI\u2010TOF\u2010MS and quantified UV spectrophotometrically using molar absorptivities calculated by an implementation of the nearest\u2010neighbors method.The central benzaldehyde residue of oligonucleotide UV melting experiments: UV melting profiles were acquired on a PerkinElmer Lambda 35 UV\u2010Vis spectrometer equipped with a Peltier temperature control unit, using quartz glass cuvettes with 10.00\u2005mm optical path length. The samples were prepared by mixing appropriate oligonucleotides (1.0\u2005\u03bcM) in 20\u2005mM cacodylate buffer (pH\u20057.4), the ionic strength of which was adjusted to 0.10\u2005M with NaClO4. Before each experiment, the samples were annealed by heating to 90\u2009\u00b0C for 30\u2005min and then allowing to gradually cool down to room temperature. Three denaturing and renaturing ramps were performed with each sample and absorbance at \u03bb=260\u2005nm was recorded at 0.5\u2009\u00b0C intervals. The melting temperatures were determined as inflection points on the UV melting curves.CD experiments: CD spectra were recorded on an Applied Photophysics Chirascan spectropolarimeter equipped with a Peltier temperature control unit, using quartz glass cuvettes with 10.00\u2005mm optical path length. Sample preparation was identical to the procedures used for the UV melting experiments above. Nine spectra were acquired for each sample at 10\u2009\u00b0C intervals over T=10\u201390\u2009\u00b0C and \u03bb=200\u2013400\u2005nm. Before each measurement, the samples were allowed to equilibrate at the appropriate temperature for either 120\u2005s or 1800\u2005s .O\u2010(tert\u2010Butyldimethylsilyl)\u20102\u2010deoxy\u20102,3\u2010didehydro\u2010\u03b2\u2010d\u2010erythro\u2010pentofuranosyl]phenyl}\u20101,3\u2010dioxane (1)2\u2010{4\u2010\u20102,3\u2010dihydrofuran\u20102\u2010yl}methanol , Bu4NBr , DIPEA and (tBu3P)2Pd(0) were taken in a round\u2010bottomed flask and purged with argon. 1,4\u2010dioxane was added and the mixture stirred at 70\u2009\u00b0C for 16\u2005h. The volatiles were evaporated and the residue was co\u2010evaporated twice from toluene. The crude product was purified by silica gel flash column chromatography affording compound 1 as a colorless foam. 1H NMR : \u03b4=7.48 , 7.39 , 5.74 , 5.50 , 4.83 , 4.62 , 4.25 , 3.97 , 3.74\u20133.66 , 2.22 , 1.95 , 1.43 , 0.96 ), 0.24 , 0.21 . 13C NMR : \u03b4=151.5 (C3\u2032), 143.0 (C1), 138.8 (C4), 127.1 (C3 & C5), 126.3 (C2 & C6), 101.4 (dioxane\u2212C1), 101.3 (C2\u2032), 84.6 (C4\u2032), 83.3 (C1\u2032), 67.3 (dioxane\u2212C3 & C5), 63.0 (C5\u2032), 25.8 (dioxane\u2212C5), 25.6 (SiC(CH3)3), 18.1 (SiC(CH3)3), \u22124.9 (SiCH3), \u22125.0 (SiCH3). HRMS (ESI+): m/z: calcd for C21H32O5SiNa [M+Na]+: 415.1911; found: 415.1894.R,5R)\u20105\u2010(Hydroxymethyl)\u20104\u2010oxotetrahydrofuran\u20102\u2010yl]phenyl}\u20101,3\u2010dioxane (2)2\u2010{4\u2010+: 301.1046; found: 301.1045.d\u2010erythro\u2010pentofuranosyl)phenyl]\u20101,3\u2010dioxane (3)2\u2010+: 303.1203; found: 303.1202.O\u2010\u20102\u2010deoxy\u2010\u03b2\u2010d\u2010erythro\u2010pentofuranosyl]phenyl}\u20101,3\u2010dioxane (4)2\u2010{4\u2010+: 605.2510; found: 605.2511.O\u2010\u20105\u2010: Et3N and 2\u2010cyanoethyl\u2010N,N\u2010diisopropylchlorophosphoramidite were added to a stirred solution of compound 4 in dry CH2Cl2 (3.0\u2005mL) at 25\u2009\u00b0C under N2. The reaction mixture was stirred for 2\u2005h, after which it was diluted with CH2Cl2 (50\u2005mL) and washed with saturated aq. NaHCO3 (50\u2005mL). The organic layer was dried over Na2SO4 and evaporated to dryness. The residue was purified by silica gel flash column chromatography affording compound 5 as a white foam. 1H NMR : \u03b4=7.52\u20137.43 , 7.41\u20137.37 , 7.32\u20137.29 , 7.24 , 6.86\u20136.83 , 5.52 , 5.19 , 4.52 , 4.30\u20134.26 , 4.01 , 3.82 , 3.74\u20133.69 , 3.67\u20133.61 , 3.36 , 3.28 , 2.47 , 2.33 , 2.26 , 2.01 , 1.47 , 1.21 2), 1.18 2). 13C NMR : \u03b4=158.5 (MeOPh\u2212C4), 144.9 (Ph\u2212C1), 142.5 (C1), 138.0 (C4), 136.2 (MeOPh\u2212C1), 136.1 (MeOPh\u2212C1), 130.18 (MeOPh\u2212C2 & C6), 130.15 (MeOPh\u2212C2 & C6), 128.3 (Ph\u2212C3 & C5), 127.8 (Ph\u2212C2 & C6), 126.7 (Ph\u2212C4), 126.0 (C3 & C5), 125.9 (C2 & C6), 117.5 (CN), 113.1 (MeOPh\u2212C3 & C5), 101.5 (dioxane\u2212C1), 86.1 (CAr3), 86.0 , 80.1 (C1\u2032), 75.7 , 67.4 (dioxane\u2212C3 & C5), 64.1 (C5\u2032), 58.3 , 55.2 (OCH3), 43.5 , 43.2 , 25.8 (dioxane\u2212C4), 24.6 2), 24.5 2), 20.2 . 31P : \u03b4=148.1. 1H NMR : \u03b4=7.51\u20137.43 , 7.40\u20137.36 , 7.30\u20137.27 , 7.22 , 6.85\u20136.81 , 5.52 , 5.19 , 4.52 , 4.29 , 4.24 , 4.01 , 3.87\u20133.77 , 3.80 , 3.62\u20133.53 , 3.31 , 3.26 , 2.63 , 2.40 , 2.25 , 1.99 , 1.47 , 1.19 2), 1.10 2). 13C NMR : \u03b4=158.4 (MeOPh\u2212C4), 144.9 (Ph\u2212C1), 142.5 (C1), 138.0 (C4), 136.13 (MeOPh\u2212C1), 136.11 (MeOPh\u2212C1), 130.17 (MeOPh\u2212C2 & C6), 130.13 (MeOPh\u2212C2 & C6), 128.3 (Ph\u2212C3 & C5), 127.8 (Ph\u2212C2 & C6), 126.7 (Ph\u2212C4), 126.0 (C3 & C5), 125.8 (C2 & C6), 117.5 (CN), 113.1 (MeOPh\u2212C3 & C5), 101.5 (dioxane\u2212C1), 86.0 (CAr3), 85.7 , 80.0 (C1\u2032), 76.1 , 67.4 (dioxane\u2212C3 & C5), 64.2 (C5\u2032), 58.4 , 55.2 (OCH3), 43.5 , 43.2 , 25.8 (dioxane\u2212C4), 24.6 2), 24.5 2), 20.4 . 31P : \u03b4=147.8. HRMS (ESI+): m/z: calcd for C45H55N2O8PK [M+K]+: 821.3328; found: 821.3325.O\u2010benzyl\u2010C\u2010phthalimidooxymethyl\u20102\u2010deoxy\u2010\u03b2\u2010d\u2010ribofuranose (7)3,5\u2010Di\u2010: A solution of DIAD in dry THF (8.0\u2005mL) was added dropwise to a stirred solution of compound [613] , N\u2010hydroxyphthalimide and PPh3 in dry THF (65.0\u2005mL) at 0\u2009\u00b0C. The resulting mixture was stirred at 0\u2009\u00b0C for 2\u2005h and then at 25\u2009\u00b0C for 12\u2005h, after which it was evaporated to dryness. The residue was suspended in H2O and extracted with EtOAc (2\u00d750\u2005mL). The combined extracts were dried over Na2SO4 and evaporated to dryness. The residue was purified by silica gel flash column chromatography affording compound 7 as a colorless syrup. 1H NMR : \u03b4=7.83 , 7.74 , 7.37\u20137.27 , 4.58 , 4.55 , 4.52 , 4.35\u20134.29 , 4.19 , 4.11 , 3.54 , 3.47 , 2.21 , 2.04 . 13C NMR : \u03b4=163.3 (C=O), 138.2 (phenyl\u2212C1), 138.1 (phenyl\u2212C1), 134.4 , 129.0 , 128.43 (phenyl\u2212C3 & C5), 128.37 (phenyl\u2212C3 & C5), 127.70 (phenyl\u2212C4), 127.68 (phenyl\u2212C4), 127.60 (phenyl\u2212C2 & C6), 127.57 (phenyl\u2212C2 & C6), 123.5 , 84.1 (C4\u2032), 80.6 (C3\u2032), 79.6 (CH2ON), 76.4 (C1\u2032), 73.4 (PhCH2), 71.2 (PhCH2), 70.9 (C5\u2032), 34.6 (C2\u2032). HRMS (ESI+): m/z: calcd for C28H27O6NNa [M+Na]+: 496.1731; found: 496.1728.Cd\u2010ribofuranose (8)\u2010phthalimidooxymethyl\u20102\u2010deoxy\u2010\u03b2\u2010: Compound 7 was dissolved in EtOAc (50.0\u2005mL). 20\u2009% Pd(OH)2/C (190.0\u2005mg) was added, the reaction vessel was filled with H2, and the reaction mixture was stirred at 25\u2009\u00b0C under H2 atmosphere for 1\u2005h. The mixture was filtered through celite, and the filter was washed with EtOAc (2\u00d720\u2005mL). The combined filtrates were concentrated under vacuum and the residue was purified by silica gel flash column chromatography affording compound 8 as colorless syrup. 1H NMR \u03b4=7.87\u20137.83 , 4.51 , 4.30\u20134.24 , 3.82 , 3.57\u20133.51 , 2.09\u20131.97 . \u03b4=163.5 (C=O), 134.4 , 128.9 , 122.9 , 87.7 (C4\u2032), 79.5 (CH2ON), 76.1 (C1\u2032), 72.1 (C3\u2032), 62.4 (C5\u2032), 36.6 (C2\u2032). m/z: calcd for C14H15O6NNa [M+Na]+: 316.0792; found: 316.0792.O\u2010\u2010C\u2010phthalimidooxymethyl\u20102\u2010deoxy\u2010\u03b2\u2010d\u2010ribofuranose (9)5\u2010: Compound 9 was prepared from compound 8 as described in the literatureO\u2010[(2\u2010Cyanoethoxy)\u2010C\u2010phthalimidooxymethyl\u20102\u2010deoxy\u2010\u03b2\u2010d\u2010ribofuranose (10)\u2010diisopropylamino)phosphinyl]\u20105\u2010: Compound 10 was prepared from compound 9 as described in the literatureThe authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "According to the literature, only small and twinned crystals of this double salt could be synthesised in a concentrated solution of MgCl2. For the crystal structure solution, single-crystal diffraction was carried out at a synchrotron radiation source. The monoclinic crystal structure (space group Cc) exhibits double chains of MgO octa\u00adhedra linked by corners, connected by carbonate units and water mol\u00adecules. The chloride ions are positioned between these double chains parallel to the (100) plane. Dry MgCO3\u00b7MgCl2\u00b77H2O decom\u00adposes in the air to chlorartinite, Mg2(OH)Cl(CO3)\u00b7nH2O (n = 2 or 3). This work includes an extensive char\u00adacterization of the title com\u00adpound by powder X-ray diffraction, thermal analysis, SEM and vibrational spectroscopy.MgCO Therefore, the system MgCl2\u2013MgCO3\u2013H2O\u2013CO2 has been investigated. The only nonbasic salt containing carbonate and chloride ions is MgCO3\u00b7MgCl2\u00b77H2O , b = 11.30\u2005(8), c = 9.22\u2005(7)\u2005\u00c5 and \u03b2\u00a0= 118.2\u2005(6)\u00b0. Due to the low scattering power and the small size of the crystals, a crystal structure analysis of single crystals was not possible until now. Our own investigations should provide a better com\u00adprehension of the synthesis of MgCO3\u00b7MgCl2\u00b77H2O and provide a more detailed characterization, including a crystal structure analysis.Schmittler 1964 conclude3\u00b7MgCl2\u00b77H2O is based on the information of Schmidt . The suspension was stirred for 30\u2005min. Afterwards, the undissolved MgO was filtered off. CO2 was bubbled through the stirred solution for 24\u2005h at room tem\u00adper\u00adature. The product was filtered off for further characterization.The synthesis of MgCOmidt 1960. MgO (1\u20053\u00b7MgCl2\u00b77H2O was recovered from a droplet of its mother liquor and mounted rapidly in the cold (150\u2005K) stream of nitro\u00adgen gas of the diffractometer.Data were collected on beamline SCD at the KIT Synchrotron Radiation Source using a Stoe IPDS diffrac\u00adtom\u00adeter with monochromated radiation of \u03bb = 0.8000\u2005\u00c5. A crystal of MgCOK\u03b11 radiation, Vantec 1 detector). The samples were prepared as flat plates and measured at room temperature.PXRD patterns were taken for phase identification with a laboratory Bruker D8 Discover powder diffractometer in Bragg\u2013Brentano set up .The thermal analysis was performed with a TG/DTA 220 instrument from Seiko Instruments (reference substance: AlThe SEM images were recorded with a TESCAN Vega 5130 SB instrument (20\u2005kV accelerating voltage). The sample was coated with gold.\u22121, 256 scans per measurement) with KBr blanks was used.For the FT\u2013IR spectrum, a Thermo Scientific Nicolet 380 FTIR spectrometer .max/\u03bb = 0.56\u2005\u00c5\u22121 could be considered for the structure refinement. The crystal structure was solved by direct methods. The resulting structure solution exhibits a chemically reasonable atomic arrangement, distances, angles and displacement parameters.Crystal data, data collection and structure refinement details are given in Table\u00a01Uiso values were set at 1.2Ueq(O) using a riding-model approximation.H atoms were placed in the positions indexed by difference Fourier maps and their The crystal exhibits nonmerohedral twinning. The matrix that relates the individual diffraction pattern was determined as . The reflections of both domains were integrated .3\u00b7MgCl2\u00b77H2O Cl(CO3)\u00b73H2O] begins within a few days 4(OH)2\u00b74H2O] and amorphous phases. At 803\u2005K the decom\u00adposition is com\u00adplete and only MgO remains in the residue. The observed mass loss of 74.3\u2005(1)% confirms the theoretical mass loss of 73.6%.The thermal decom\u00adposition of MgCOps Fig.\u00a02. H2O, COmann 1964. A preci3\u00b7MgCl2\u00b77H2O show thin needles (50 \u00d7 5\u2005\u00b5m), which are twinned or even more inter\u00adgrown Cl(CO3)\u00b73H2O plane, are located the chloride ions Cl1 and Cl2 Fig.\u00a08. The poss Table\u00a03. As a co3\u00b7MgCl2\u00b77H2O and MgCO3\u00b73H2O Cl(CO3)\u00b72H2O (chlorartinite) and Mg2(OH)Cl(CO3)\u00b7H2O (dehydrated clorarti\u00adnite), do not exhibit such double chains I. DOI: 10.1107/S2053229620008153/uk3196Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2053229620008153/uk3196Isup3.cmlSupporting information file. DOI: 2010753CCDC reference:"} +{"text": "In the ferrocene unit, the penta\u00addienyl (Cps) rings are in an eclipsed conformation. Strong inter\u00admolecular \u03c0\u2013\u03c0-stacking, CH(Cp)\u2014C(Cp), and O(p-nitro\u00adphen\u00adyl)\u2014HC(Cp) inter\u00adactions consolidate the crystal structure.The title ferrocene derivative including an alkyne and a 5H5)2(C8NO2)], including an alkyne bonded to a para-nitro\u00adphenyl substituent, which was synthesized from a copper-free Sonogashira cross-coupling reaction between ethynylferrocene and 4-bromo-1-nitro\u00adbenzene, crystallizes in the P21/n space group. In the ferrocene unit, the penta\u00addienyl (Cps) rings are in an eclipsed conformation. The angle of rotation between the substituted cyclo\u00adpenta\u00addienyl ring and the p-nitro\u00adphenyl group is 6.19\u2005(10)\u00b0, yielding a quasi-linear extension of the ferrocenyl substitution. Important inter\u00admolecular inter\u00adactions arise from \u03c0\u2013\u03c0 stacking between the Cp rings and the p-nitro\u00adphenyl, from corners of the Cp rings that are perpendicularly aligned, and between the O atoms from the nitro substituent and carbons at the corners of the Cp rings, propagating along all three crystallographic axes.The title ferrocene derivative, [Fe(C The distances of the Fe1 atom from the centroids of the substituted and unsubstituted Cp rings are 1.6461\u2005(8) and 1.6584\u2005(8)\u2005\u00c5, respectively. The Cg1\u2014Fe1\u2014Cg2 angle is 179.27\u00b0, where Cg1 and Cg2 are the centroids of substituted and unsubstituted Cp rings, respectively. The Cp rings in the ferrocene system are thus almost parallel, since the angle between the Cp ring planes is 1.03\u2005(13)\u00b0. In addition, the Cp rings display a nearly eclipsed conformation with a slight deviation, as demonstrated by the average C\u2014Cg1\u2014Cg2\u2014C torsion angle of 12.26\u00b0. The C\u2014C bond distances in the Cp rings range from 1.417\u2005(2) to 1.436\u2005(2)\u2005\u00c5, while the Fe\u2014C bond lengths range between 2.038\u2005(2) and 2.055\u2005(2)\u2005\u00c5.Fig.\u00a01p-nitro\u00adphenyl substituent, allowing the formation of a zigzag structure; atom pairs involved relate C6(Cp) and C7(Cp) to C17(p-nitro\u00adphen\u00adyl) and C18(p-nitro\u00adphen\u00adyl) of a neighboring mol\u00adecule, with short contacts of 3.340\u2005(2) and 3.397\u2005(2)\u2005\u00c5, respectively. This inter\u00adaction can be described as pairs of mol\u00adecules being inter\u00adrupted by two C3(Cp)\u22efH8\u2014C8(Cp) inter\u00adactions from a different inter\u00adconnected pair of perpendicularly oriented Fc moieties with short contact distances of 2.83\u2005\u00c5 each. Short contacts from neighboring mol\u00adecules establishing a distinctive inter\u00adconnected pair between a corner of the Cp ring and one of the oxygen atoms from the p-nitro\u00adphenyl substituent yield a closed arrangement of atoms. Short contacts involve H6\u2014C6(Cp)\u22efO1(p-nitro\u00adphen\u00adyl) at a distance of 3.461\u2005(2)\u2005\u00c5. Another inter\u00adconnection is found between adjacent p-nitro\u00adphenyl groups, yielding a ring arrangement involving pairs from H17\u2014C17(p-nitro\u00adphen\u00adyl)\u22efO2(p-nitro\u00adphen\u00adyl) with a distance of 2.727\u2005(2)\u2005\u00c5 and pairs from O1(p-nitro\u00adphen\u00adyl)\u22efH15\u2014C15(p-nitro\u00adphen\u00adyl) with a distance of 2.716\u2005(2)\u2005\u00c5. In addition, a chain is formed by short contacts from the C17\u2014H17(p-nitro\u00adphen\u00adyl)\u22efO1(p-nitro\u00adphen\u00adyl) inter\u00adaction belonging to the p-nitro\u00adphenyl substituent with a distance of 3.203\u2005(19)\u2005\u00c5. Numerical details of the hydrogen-bonding inter\u00adactions are given in Table\u00a01The title compound exhibits \u03c0\u2013\u03c0 stacking inter\u00adactions between one of the Cp rings from the Fc moiety and the CrystalExplorer17 and those decomposed into nine individual inter\u00adactions: H\u22efH , C\u22efH/ H\u22efC , O\u22efH/ H\u22efO , C\u22efC , C\u22efO/O\u22efC , C\u22efN/N\u22efC , N\u22efO/O\u22efN , O\u22efO and N\u22efH/H\u22efN . The Hirshfeld surface analysis for the title compound indicates that the most significant contributions arise from H\u22efH and C\u22efH contacts 2 (0.01\u2005mmol), Et3N (2\u2005mmol) and 4-bromo-1-nitro\u00adbenzene (1.0\u2005mmol) to a 25\u2005mL round-bottom flask, followed by the addition of DMF (1.0\u2005mL) by syringe. The reaction was stirred for 1\u2005h at 353\u2005K. The reaction was stopped and crashed out with 20\u2005mL of cold distilled water, then the solid was vacuum filtrated, and chromatographed [silica (hepta\u00adne\u2013ethyl acetate/7:3)] to afford the pure compound, 70% yield. Dark-red crystals suitable for X-ray diffraction were obtained by the slow evaporation of CDCl3 solution of the title compound at room temperature. NMR analyses were performed on a Bruker AV-700 spectrometer by using CDCl3 99.9% pure as a solvent and Me4Si as external standard.1H NMR : 4.26 , 4.32 , 4.55 , 7.59 , 8.18 . 13C NMR : 63.6, 69.6, 70.1, 71.8, 84.5, 95.2, 123.6, 131.1, 131.8, 146.4. IR : 2200 (C\u2261C). Electrochemistry: .The title compound was prepared by adding ethynylferrocene (1.0\u2005mmol), PdClUiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989020010336/dj2003sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020010336/dj2003Isup2.hklStructure factors: contains datablock(s) I. DOI: 1991635CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Figure\u00a06 Liver control [1] appears identical to Fig.\u00a010 liver control [2]Figure\u00a06 Lung PEG-NPs [1] appears identical to Fig.\u00a010 lung PEG-CF [2]Figure\u00a06 Spleen control [1] appears identical to Fig.\u00a010 spleen control [2]Figure\u00a06 Spleen treatment [1] appears identical to Fig.\u00a010 spleen treatment [2]Figure 7A [1] appears to be identical but in a different order to Fig.\u00a06A all panels [3]Figure 7B [1] appears identical to Fig.\u00a06B [3]Figure 7D [1] appears identical to Fig.\u00a06D [3] using a different scaleFigure 7C [1] appears identical to Fig.\u00a06E [3] using a different scaleAll authors agree with this retraction.The authors have retracted their article because"} +{"text": "Type 1 diabetes (T1D) and celiac disease (CD) share common genetic loci, mainly within the human leukocyte antigen (HLA) class II complex. Extended genotyping of HLA class II alleles and their potential risk for developing both diseases remains to be studied. The present study compared extended HLA\u2010class II gene polymorphisms in children with T1D, CD, and a subgroup diagnosed with both diseases (T1D w/CD).Next\u2010generation targeted sequencing (NGTS) of HLA\u2010DRB3, DRB4, DRB5, DRB1, DQA1, DQB1, DPA1, and DPB1 alleles from DNA collected from 68 T1D, 219 CD, and seven T1D w/CD patients were compared with 636 HLA\u2010genotyped Swedish children from the general population selected as controls.DRB4*01:03:01 allele occurred more frequently in T1D w/CD (odds ratio (OR)\u2009=\u20097.84; 95% confidence interval (95% CI)\u2009=\u2009, P\u2009=\u20090.0002) and T1D , P\u2009=\u20091.07\u2009\u00d7\u200910\u221214), respectively. The DRB3*01:01:02 allele occurred more frequently in CD as compared to controls , P\u2009=\u20094.24\u2009\u00d7\u200910\u221271), but less frequently in T1D , P\u2009=\u20097.29\u2009\u00d7\u200910\u221207) and T1D w/CD , P\u2009\u2264\u20090.999). The frequency of the DRB4*01:03:01\u2010DRB1*04:01:01\u2010DQA1*03:01:01\u2010DQB1*03:02:01 (DR4\u2010DQ8) haplotype was higher in T1D w/CD P\u2009=\u20093.75\u2009\u00d7\u200910\u22129), and moderately higher in T1D P\u2009=\u20090.01) compared with controls, but comparable in CD , P\u2009=\u20090.08) and controls.In comparison to controls, the DRB4*01:03:01, DRB3*01:01:02, and DRB3*02:02:01 of which DRB4*01:03:01 confers the strongest risk allele for developing T1D w/CD.Children with T1D and CD are associated with HLA\u2010class II allele sequencing was performed using the ScisGo HLA v4 typing kit . Known haplotypes were used to phase the extended haplotypes and to predict the genotypes. Amplicon\u2010based 2\u2010stage polymerase chain reaction (PCR)\u2010based amplification of HLA\u2010class II alleles and sequencing\u2010by\u2010synthesis approach by using fluorescently labeled reversible terminator nucleotides with MiSeq v2 PE500 technology was performed as previously described.2.3\u03c72 tests and Fisher's exact tests (if any cell contained fewer than three observations) were used to test whether the frequencies of a given allele/haplotype differed between cases and noncases. The RPE method was used to identify the disease risk alleles, haplotypes or genotypes with the strongest predisposing or protective effects at each iteration. The selected alleles were then removed from the dataset, and the analysis was repeated until no risk or protective alleles were identified. Comparisons of DR and DQ allele frequencies were performed for both exons 2 and 3 of chromosome 6p21 by performing pairwise comparisons between all study groups and listed in order of increasing P\u2010value, followed by the extended haplotype and genotype frequencies. In the HLA\u2010DR locus, alleles of HLA\u2010DRB1 are in linkage disequilibrium with alleles of either HLA\u2010DRB3 or DRB4 or DRB5. Hence, by treating these highly dependent DR subtypes and their allelic variations as different alleles, analysis of HLA\u2010DRB1 and the secondary DRB3, DRB4 or DRB5 alleles by PRE method produces estimates of frequencies for all haplotypes provided that expectation numbers of corresponding haplotypes are five or more copies in both patients and control subjects. P\u2010values \u2264.05 were considered statistically significant, and alleles with a low frequency (\u22641%) were not shown in the analysis. The P\u2010values presented are nominal and not adjusted for multiple comparisons. Analyses were performed in R (r-project.org) version 3.6.1 and R package epiDisplay version 3.5.0.1.Allelic frequency distribution analysis of HLA\u2010class II genes was performed using the relative predispositional effects (RPE) analysis.33.1DRB345, \u2010DRB1, \u2010DQA1 and \u2010DQB1 allele frequencies in children with CD compared with controls are summarized in Table S1. The allelic distributions of the disease associated DRB alleles differed significantly between all the patient groups and the controls , P\u2009=\u20094.24\u2009\u00d7\u200910\u221271), DRB4*01:03:01 in 32% , P\u2009=\u20090.002) and DRB4*01:03:02 in only 1.4% , P\u2009=\u20090.021). Likewise, DRB3*02:02:01 occurred in 4.6% which was less frequent than in controls , P\u2009=\u20091.23\u2009\u00d7\u200910\u22129). DRB1*03:01:01 occurred in 64% of CD children which was higher than in the controls , P\u2009=\u20093.82\u2009\u00d7\u200910\u2212105). The DRB1*04:01:01 allele was found in 19% P\u2009=\u20091.85\u2009\u00d7\u200910\u22127) and DRB1*04:04:01 found in 12% , P\u2009=\u20095.66\u2009\u00d7\u200910\u22128). The DRB1*08:01:01 allele had a protective effect in 1% , P\u2009=\u20090.0005).HLA\u2010ls Table and 3. ADQA1*05:01:01 occurred in 64% of the CD children , P\u2009=\u20093.82\u2009\u00d7\u200910\u2212105), DQA1*03:01:01 in 31% , P\u2009=\u20092.92\u2009\u00d7\u200910\u221218) and DQA1*03:02:01 in 2.5% , P\u2009=\u20090.002). The DQB1*02:01:01 allele occurred in 54% of the CD children , P\u2009=\u20092.06\u2009\u00d7\u200910\u2212109) and DQB1*03:02:01 in 33% , P\u2009=\u20091.46\u2009\u00d7\u200910\u221222). DQB1*04:02:01 occurred in 1% , P\u2009=\u20090.0001).Among the HLA\u2010DQA1 and DQB1 alleles, DRB3*01:01:02\u2010DRB1*03:01:01\u2010DQA1*05:01:01\u2010DQB1*02:01:01 (DRB3\u2010DR3\u2010DQ2.5) was found in 46% of CD children , P\u2009=\u20091.12\u2009\u00d7\u200910\u221261) and DRB3*02:02:01\u2010DRB1*03:01:01\u2010DQA1*05:01:01\u2010DQB1*02:01:01 in 3% , P\u2009=\u20090.0496). On a genotype level, a total of 29 different haplotype combinations were found in CD children compared with 315 in controls. Being homozygous for DRB3*01:01:02\u2010DRB1*03:01:01\u2010DQA1*05:01:01\u2010DQB1*02:01:01(DR3\u2010DQ2.5/DR3\u2010DQ2.5) was the most significant genotype (43) of CD group when compared to the control group , P\u2009=\u20091.42\u2009\u00d7\u200910\u221257, Data not shown).A complete list of HLA\u2010DRB\u2010DQA1\u2010DQB1 haplotypes is summarized in Table S4. Of these, only two haplotypes were observed more frequently in CD children compared with controls: 3.2DRB345, \u2010DRB1, \u2010DQA1 and \u2010DQB1 allele frequencies in T1D children and controls are summarized in Table S2. Of these alleles, DRB4*01:03:01 was found in 55% of T1D children , P\u2009=\u20091.07\u2009\u00d7\u200910\u221214), DRB3*01:01:02 in 34% , P\u2009=\u20097.29\u2009\u00d7\u200910\u221207), DRB3*02:02:01 in 4.4%, which were less frequent compared with controls , P\u2009=\u20090.0003). The DRB1*04:01:01 allele was the most frequent DRB1 allele and occurred in 39% , P\u2009=\u20094\u2009\u00d7\u200910\u221222), followed by DRB1*03:01:01 in 38% , P\u2009=\u20091.05\u2009\u00d7\u200910\u221215), and DRB1*04:04:01 in 11% , P\u2009=\u20090.002). The DQA1*03:01:01 allele occurred in 52% of T1D children , P\u2009=\u20095.38\u2009\u00d7\u200910\u22123) and DQA1*05:01:01 in 38% , P\u2009=\u20091.05\u2009\u00d7\u200910\u221215). DQB1*03:02:01 was present in 54% , P\u2009=\u20091.29\u2009\u00d7\u200910\u221235) and DQB1*02:01:01 in 37.5% , P\u2009=\u20094.43\u2009\u00d7\u200910\u221217). DPB1*04:02:01 occured in 2.2%, which was less frequent compared with controls , P\u2009=\u20090001). In contrast, DPB1*01:01:01 was associated with increased risk , P\u2009=\u20090007).HLA\u2010DRB4*01:03:01\u2010DRB1*04:01:01\u2010DQA1*03:01:01\u2010DQB1*03:02:01 (DRB4\u2010DR4\u2010DQ8) was more frequently found in T1D children (11%) compared with controls , P\u2009=\u20090.01). On a genotype level, 20 different different haplotype combinations were identified. Among those, DRB3*01:01:02\u2010DRB1*03:01:01\u2010DQA1*05:01:01\u2010DQB1*02:01:01(DR3\u2010DQ2.5/DR3\u2010DQ2.5) was found in 4.4% of T1D children , P\u2009=\u20090.0339, Data not shown).Haplotype frequencies in T1D children are summarized in Table S5. Only 3.3DRB4*01:03:01 was the most frequent occurring allele and found in 71.4% of T1D w/CD children , P\u2009=\u20090.0002), in comparison to 14.3% that carried either DRB3*01:01:02 or DRB3*02:02:01(P \u2264 0.999). DRB1*04:01:01 was present in 64% of T1D w/CD , P\u2009=\u20091.53\u2009\u00d7\u200910\u221206), DRB1*03:01:01 in 29% , P\u2009=\u20090.08). DQA1*03:01:01 was present in 57% of T1D w/CD children , P\u2009=\u20091.07\u2009\u00d7\u200910\u221206) and DQA1*05:01:01 in 29% . DQB1*03:02:01 occurred in 71% of T1D w/CD children , P\u2009=\u20097.95\u2009\u00d7\u200910\u221207) and DQB1*02:01:01 in 29% , P\u2009=\u20090.06). Haplotype distributions are listed in Table S6. Among those, only DRB4*01:03:01\u2010DRB1*04:01:01\u2010DQA1*03:01:01\u2010DQB1*03:02:01 occurred more frequently in T1D w/CD children compared with controls and found in 42.9% , P\u2009=\u20093.75\u2009\u00d7\u200910\u22129). Four different DRB3 DRB4 DRB5\u2010DRB1\u2010DQA1\u2010DQB1 genotypes were found among T1D w/CD children, but there was no difference compared with controls.HLA class II alleles frequencies are summarized in Table S3. 3.4DRB3, \u2010DRB4, \u2010DRB5 alleles using RPE analysis (Table S1\u2010S2\u2010S3), DRB4*01:03:01 rank higher in T1D and T1D w/CD predisposition than DRB3*01:01:02, which rank higher in CD predisposition in comparison to the control group. DRB4*01:03:01 occurred in 55% of T1D children , P \u2009=\u20091.59\u2009\u00d7\u200910\u221206) and 71% of T1D w/ CD children , P\u2009=\u20090.003), which was higher compared with 32% of CD children. The opposite was true for DRB3*01:01:02, which was found in 60% of CD children compared with 33% of T1D children , P\u2009=\u20097.17\u2009\u00d7\u200910\u221209) , P\u2009=\u20096.3\u2009\u00d7\u200910\u201006) and 64% of T1D w/CD children , P\u2009=\u20090.0004) compared with only 19% of CD children, whereas DRB1*03:01:01 was found in 66% of CD children compared with 37% in T1D , P\u2009=\u20093.28\u2009\u00d7\u200910\u221209) and 28% of T1D w/CD , P\u2009=\u20090.009), respectively , P\u2009=\u20093.28\u2009\u00d7\u200910\u221209) and 28% of T1D w/CD , P\u2009=\u20090.009), respectively. In contrast, DQB1*03:02:01 was more frequently found in T1D (54%) but not compared with CD (34%) , P\u2009=\u20092.36\u2009\u00d7\u200910\u221205) compared with 71% of T1D w/CD , P\u2009=\u20090.007).By estimating the hierarchy of risk among all \u2010DRB3*01:01:02\u2010DRB1*03:01:01\u2010DQA1*05:01:01\u2010DQB1*02:01:01 (DRB3\u2010DR3\u2010DQ2.5) was the most common haplotype and occurred in 45% of CD children compared with 5% in T1D (P\u2009=\u20095.89\u2009\u00d7\u200910\u221218), while DRB3*01:01:02\u2010DRB1*03:01:01\u2010DQA1*03:01:01\u2010DQB1*02:01:01 was the most common haplotype in T1D. The DRB4*01:03:01\u2010DRB1*04:01:01\u2010DQA1*03:01:01\u2010DQB1*03:02:01 (DR4\u2010DQ8) haplotype was carried in 42% of T1D w/CD children compared with 11% in T1D , P\u2009=\u20090.001) and 7.8% in CD , P\u2009=\u20095.33\u2009\u00d7\u200910\u221206) children was the most frequent genotype and found in 43% of CD children compared with 4.4% in T1D , P\u2009=\u20097.92\u2009\u00d7\u200910\u221211, Data not shown). In T1D, the DRB3*01:01:02\u2010DRB1*03:01:01\u2010DQA1*03:01:01\u2010DQB1*02:01:01/DRB4*01:03:01\u2010DRB1*04:01:01\u2010DQA1*05:01:01\u2010DQB1*03:02:01 (DR3\u2010DQ2/DR4\u2010DQ8) genotype was found in 38% of T1D children compared with 17% of CD children , P\u2009=\u20090.0002) and 28% of children with T1D w/CD . In T1D w/CD, being homozygous for DRB4*01:03:01\u2010DRB1*04:01:01\u2010DQA1*05:01:01\u2010DQB1*03:02:01 (DR4\u2010DQ8/DR4\u2010DQ8) was the most frequent genotype and found in 42% , P\u2009=\u20090.002) compared with 3.2% in CD and 10% in T1D , P\u2009=\u20090.04, Data not shown) children, respectively.In CD children, the 4DRB4*01:03:01, DRB3*01:01:02, and DRB3*02:02:01 alleles, were found to be associated with T1D w/CD. Of these three alleles DRB4*01:03:01 was associated with T1D only, DRB3*01:01:02 was associated with CD only, but inversely associated with T1D only. The DRB3*02:02:01 allele was associated with a low predisposition in all three groups. The novelty of the study lies in the fact that several alleles in DRB3, DRB4 and DRB5 among T1D w/CD children seem to have an extended HLA polymorphism more similar to that in children with T1D than that in children with CD.The present study aimed to fill the gap of knowledge of why some children with either T1D or CD are at an increased risk for developing both diseases (T1D w/CD) by analyzing extended HLA class II genes. The main findings were that the DRB4*01:03:01\u2010containing haplotypes conferred a positive association with T1D w/CD as well as T1D in comparison to DRB3*01:01:02\u2010containing haplotypes that were positively associated with CD. Around 50% of T1D and 70% of T1D w/CD haplotypes carry the former allele compared with 60% of CD haplotypes which carry the latter allele. Noting that all copies of chromosome 6 have a DRB1 locus, and most, but not all, have a functional second DRB locus, DRB4 was shown to be secondary for DRB1*04 haplotypes. The increase of DRB4*01:03:01 on DRB1*04:01 haplotypes in DRB1*04:01/*04:01 T1D w/CD case subjects versus DRB1*03:01/DRB1*04:01 case subjects and control subjects could reflect linkage of disequilibrium (LD) with alleles at other high\u2010risk loci. The analyses of the extended DRB1\u2010DRB3\u2010DRB4\u2010DRB5 haplotypes suggests that the risk for developing both diseases likely resembles T1D risk.Dissecting the extended DRB1\u2010DRB3\u2010DRB4\u2010DRB5 haplotypes, On a genotype level, differences between T1D w/CD and T1D children were found of whom T1D w/CD children were more likely to be homozygous for DR4\u2010DQ8/DR4\u2010DQ8 compared with T1D children. In line with other studies, the DR3\u2010DQ2 haplotype occurred in over a third of T1D children,trans, in addition to the DQ molecules encoded by alleles in cis, in linkage disequilibrium; DQA1*05, and DQB1*02, encoding the DQ2.5 molecule, and DQA1*03 and DQB1*03, encoding the DQ8 molecule.The suggested hypothesis for the increased susceptibility to CD and T1D coexistence is the putative presence of DQ heterodimers encoded by alleles in The strengths of the present study were the use of high\u2010resolution NGTS for extended HLA genotyping, which enabled examining the disease susceptibility between T1D w/CD and extended HLA\u2010DRB3, DRB4 and DRB5 alleles. The RPE analysis used to estimate the association between HLA alleles (or haplotypes) and each of the outcomes accounts for the fact that a high frequency of a given allele \u201cinduces\u201d a lower frequency of all other alleles, as their total must remain constant.DRB4*01:03:01 and DRB3*01:01:02 of which DRB4*01:03:01 conferred the strongest risk allele for developing T1D w/CD.In conclusion, NGTS for genetic risk profiling of children with T1D and CD showed shared risk associations with The authors have declared no conflicting interests.Table S1 RPE analysis of the estimated frequencies of HLA\u2010DRB3, DRB4, DRB5, DRB1, DQA1 and DQB1 alleles among celiac patients and population controls. (XLSX)Table S2 RPE analysis of the estimated frequencies of HLA\u2010DRB3, DRB4, DRB5, DRB1, DQA1 and DQB1 alleles among type 1 diabetes patients and population controls. (XLSX)Table S3 RPE analysis of the estimated frequencies of HLA\u2010DRB3, DRB4, DRB5, DRB1, DQA1 and DQB1 alleles among type 1 diabetes with celiac disease patients and population controls. (XLSX)Table S4 RPE analysis of estimated frequencies of HLA\u2010DRB3, DRB4, DRB5, DRB1, DQA1 and DQB1 haplotypes among celiac patients and population controls. (XLSX)Table S5 RPE analysis of estimated frequencies of HLA\u2010DRB3, DRB4, DRB5, DRB1, DQA1 and DQB1 haplotypes among type 1 diabetes and population controls. (XLSX)Table S6 RPE analysis of the estimated frequencies of HLA\u2010DRB3, DRB4, DRB5, DRB1, DQA1 and DQB1 haplotypes among type 1 diabetes with celiac disease and population controls. (XLSX)Click here for additional data file."} +{"text": "Kampert et al. express questions regarding (1) the measurement of pulmonary function, (2) listing of the data, (3) and a model of blood flow redistribution at peak exercise described by Harms et al. In addition, (4) they comment on the peak metabolic demand of the three conditions.p = 0.001, tidal volume sm \u22129.9\u2009\u00b1\u200911.3% and ffpm: \u221214.4\u2009\u00b1\u200913.0%, p = 0.016, inhalation time sm: +12\u2009\u00b1\u200915%, ffpm: +19\u2009\u00b1\u200916%, p < 0.001, compared to nm.Ad 1: During CPET, VE sm: \u221212.0\u2009\u00b1\u200912.6%, ffpm: \u221223.1\u2009\u00b1\u200913.6%, Ad 2: The complete results are listed in Tables 2 and 3.Ad 3: Harms et al. 1998) were cited describing possible changes in the distribution of total blood flow with additional work of breathing (e.g. higher breathing resistance) [998 were p\u00a0=\u00a00.596).Ad 4: At maximum load, the metabolic demands were similar in all three conditions. The respiratory exchange ratio (RER) did not differ between the tests (nm 1.13\u00a0\u00b1\u00a00.08, sm 1.15\u00a0\u00b1\u00a00.09, ffpm 1.13\u00a0\u00b1\u00a00.08, one-way ANOVA A straightforward way to get a feeling for the clear results of the study could be to personally try out the effects of sm and ffpm during exercise."} +{"text": "The structure of the title compound exhibits a folded conformation with the three arms all on the same side of the tertiary N atom. The crystal packing features \u03c0\u2013\u03c0 inter\u00adactions. 29H26N4O6, exhibits a folded conformation with the three arms all on the same side of the tertiary N atom. The two phthalimide units make a dihedral angle of 12.18\u2005(12)\u00b0 and the dihedral angles between the benzyl plane and the phthalimide units are 68.08\u2005(7) and 67.71\u2005(7)\u00b0. The crystal packing features \u03c0\u2013\u03c0 inter\u00adactions.The structure of the title compound, C There is also a longer centrosymmetric interaction of the nitro benzyl groups (N4/C24\u2013C29) with a distance of 4.694\u2005(5)\u2005\u00c5.The crystal structure consists of centrosymmetrical dimers with off-set \u03c0\u2013\u03c0 stacking between phthalimide groups (N3/C15\u2013C22) running along the on Fig.\u00a02. The cenet al., 20162CH2CH2N)3 resulted in 149 entries. Similar off-set \u03c0\u2013\u03c0 stacking was seen in another compound with two phthalimide groups . 3,3\u2032-Diphthal\u00adimido\u00addi\u00adpropyl\u00adamine , 2-nitro\u00adbenzyl\u00adchloride , and potassium carbonate were heated at 433\u2005K for one\u2005h to give the title compound. Crystals suitable for X-ray analysis were slowly grown from chloro\u00adform.The title compound was prepared by using a previously reported method (Keypour Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2056989020016771/zn2003sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020016771/zn2003Isup4.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020016771/zn2003Isup3.cmlSupporting information file. DOI: 2052908CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure of the title compound, mol\u00adecules are linked through C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming chains parallel to the [010] direction. The mol\u00adecular geometry in the ground state was been calculated using DFT. Additionally, frontier mol\u00adecular orbital and mol\u00adecular electrostatic potential map analyses were performed. 14H12ClNO, the mol\u00adecules are linked through C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming chains parallel to the [010] direction. \u03c0\u2013\u03c0 inter\u00adactions and intra\u00admolecular hydrogen bonds are also observed. The mol\u00adecular geometry of the title compound in the ground state has been calculated using density functional theory at the B3LYP level with the 6\u2013311++G basis set. Additionally, frontier mol\u00adecular orbital and mol\u00adecular electrostatic potential map analyses were performed.In the crystal structure of the title compound, C The mol\u00adecular structure has two planar rings. The whole mol\u00adecule is approximately planar, with a maximum deviation of \u22120.0236\u2005(12)\u2005\u00c5 from planarity for the C8 atom of Schiff base. The title compound displays an E configuration with respect to the C8=N double bond. The dihedral angle between the two phenyl ring planes is 0.34\u2005(9)\u00b0 and the C5\u2014C8\u2014N1\u2014C9 torsion angle is \u2212179.81\u2005(15)\u00b0. The planar mol\u00adecular conformation is stabilized by the intra\u00admolecular O1\u2014H1\u22efN1 hydrogen bond, which forms an S(6) motif.The structure of the title compound (I)In the crystal, the mol\u00adecules are linked by C10\u2014H10\u22efO1 hydrogen bonds Table\u00a01, generaton Fig.\u00a02. C\u2014H\u22ef\u03c0 ion Fig.\u00a02. \u03c0\u2013\u03c0 staet al., 2016E)-2-{[(3-chloro\u00adphen\u00adyl)imino]\u00admeth\u00adyl}-6-methyl\u00adphen\u00adol structure. With a value of 1.271\u2005(2)\u2005\u00c5, the N1\u2014C8 bond in the title compound (I)E)-(5-chloro-2-methyl\u00adphen\u00adyl)imino\u00admeth\u00adyl]-4-methyl\u00adphenol -imino\u00admeth\u00adyl]phenol salicylaldimine -2-[(3-chloro\u00adphenyl\u00adimino)\u00admeth\u00adyl]-4-meth\u00adoxy\u00adphenol -[imino]\u00admeth\u00adyl}benzene-1,2-diol salicylaldimine level and ionization potential (IP) can be defined as A = \u2212ELUMO and IP = \u2212EHOMO. Additionally, these values can also be used calculate the electronegativity (\u03c7), chemical hardness (\u03b7) and chemical softness (S) method. In Fig.\u00a05The analysis of a three-dimensional plot of the mol\u00adecular electrostatic potential (MEP) surface is a technique for mapping the electrostatic potential onto the isoelectronic density surface, providing information about the reactive sites. The surface simultaneously displays mol\u00adecular size and shape and the electrostatic potential value. In the colour scheme adopted, red indicates an electron-rich region with a partial negative charge and blue an electron-deficient region with partial positive charge, light blue indicates a slightly electron-deficient region, yellow a slightly electron-rich region and green a neutral region and 4-chloro\u00adaniline was stirred with ethanol (30\u2005mL) at 377\u2005K for 4\u2005h, affording the title compound . Single crystals suitable for X-ray measurements were obtained by recrystallization from methanol at room temperature.Uiso(H) = 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019017353/mw2154sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019017353/mw2154Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019017353/mw2154Isup3.cmlSupporting information file. DOI: 1974885CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Pbca. The phenol ring is inclined to the benzo\u00adnitrile ring by 25.65\u2005(3)\u00b0. The configuration about the C=N bond is E, stabilized by a strong intra\u00admolecular O\u2014H\u22efN hydrogen bond that forms an S(6) ring motif.The title compound crystallizes in the ortho\u00adrhom\u00adbic space group 15H12N2O, was synthesized by condensation reaction of 2-hy\u00addroxy-5-methyl\u00adbenzaldehyde and 2-amino\u00adbenzo\u00adnitrile, and crystallizes in the ortho\u00adrhom\u00adbic space group Pbca. The phenol ring is inclined to the benzo\u00adnitrile ring by 25.65\u2005(3)\u00b0. The configuration about the C=N bond is E, stabilized by a strong intra\u00admolecular O\u2014H\u22efN hydrogen bond that forms an S(6) ring motif. In the crystal, C\u2014H\u22efO and C\u2014H\u22efN inter\u00adactions lead to the formation of sheets perpendicular to the a axis. C\u2014H\u22ef\u03c0 inter\u00adactions, forming polymeric chains along the a-axis direction, connect these sheets into a three-dimensional network. A Hirshfeld surface analysis indicates that the most important contributions for the packing arrangement are from H\u22efH and C\u22efH/H\u22efC inter\u00adactions. The density functional theory (DFT) optimized structure at the B3LYP/6\u2013311\u2005G level is compared with the experimentally determined mol\u00adecular structure and the HOMO\u2013LUMO energy gap is given.The title compound, C RCH=N\u2013R\u2032) are prepared by a condensation reaction between amines and reactive carbonyl compounds, such as aldehydes. Schiff bases are employed as catalyst carriers and keto\u2013amine (NH) forms. Depending on the tautomers, two types of intra\u00admolecular hydrogen bonds are generally observed in ortho-hy\u00addroxy Schiff bases, namely O\u2014H\u22efN in enol\u2013imine and N\u2014H\u22efO in keto\u2013amine tautomers ring motif . The C10\u2014O1 bond length [1.3503\u2005(17)\u2005\u00c5 for X-ray and 1.337\u2005\u00c5 for B3LYP] indicates single-bond character, while the imine C8=N2 bond length [1.2795\u2005(17)\u00c5 for X-ray and 1.291\u2005\u00c5 for B3LYP] indicates double-bond character. All bond lengths and bond angles are within normal ranges and are comparable with those in related Schiff base compounds . As shown in Fig.\u00a05b, the most widely scattered points in the fingerprint plot are related to H\u22efH contacts, which make a contribution of 39.2% to the Hirshfeld surface. There are also N\u22efH/H\u22efN , O\u22efH/H\u22efO and C\u22efC contacts, with smaller contributions from C\u22efN/N\u22efC (2.6%), C\u22efO/O\u22efC (0.4%) and N\u22efN (0.3%) contacts.In order to better visualize and analyze the role of weak inter\u00admolecular contacts in the crystal, a Hirshfeld surface (HS) analysis calculations using the standard B3LYP functional and a 6-311G basis-set , the ionization potential (I = -ELUMO), HOMO\u2013LUMO energy gap (\u0394E), the chemical hardness (\u03b7) and softness (S) of the title compound were predicted based on the EHOMO and ELUMO energies and the lowest-unoccupied mol\u00adecular orbital (LUMO) are very important parameters for quantum chemistry. Many electronic, optical and chemical reactivity properties of compounds can be predicted from frontier mol\u00adecular orbitals -2-{[(3-chloro\u00adphen\u00adyl)imino]\u00admeth\u00adyl}-6-methyl\u00adphenol -2-[(2-hy\u00addroxy-5-meth\u00adoxy\u00adbenzyl\u00adidene)amino]\u00adbenzo\u00adnitrile -2-[(5-bromo-2-hy\u00addroxy\u00adbenzyl\u00adidene)amino]\u00adbenzo\u00adnitrile \u00adbenzo\u00adnitrile -2-\u00adbenzo\u00adnitrile -2-(4-di\u00adethyl\u00adamino-2-hy\u00addroxy\u00adbenzyl\u00adidene\u00adamino)\u00adben\u00adzo\u00adnitrile -2-[amino]\u00adbenzo\u00adnitrile ring motif. The dihedral angles between the aromatic rings are generally smaller than the value of 25.65\u2005(3)\u00b0 observed for the title compound, with angles between 1.09\u2005(4)\u00b0 (for FOWXOF and GEJGAE) and 13.84\u2005(13)\u00b0 (for PUJDOO). Only YOVBUH features angles similar to those of (I)A search of the Cambridge Structural Database in ethanol (15\u2005ml) and 2-amino\u00adbenzo\u00adnitrile in ethanol (15\u2005ml) and stirring the mixture for 5\u2005h under reflux . Single crystals of the title compound suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution.Uiso(H) = 1.2\u20131.5Ueq(C). The position of the H1 atom was obtained from a difference map; it was placed in a calculated position with a fixed C\u2014O\u2014H angle, but the O\u2014H distance and the torsion angle were allowed to freely refine.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989020008907/zl2787sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020008907/zl2787Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020008907/zl2787Isup3.cmlSupporting information file. DOI: 2013269CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The structures of the four isomeric hydrogen-bonded 1:1 co-crystals of 6-methyl\u00adquinoline with 2-chloro-4-nitro\u00adbenzoic acid, 2-chloro-5-nitro\u00adbenzoic acid, 3-chloro-2-nitro\u00adbenzoic acid and 4-chloro-2-nitro\u00adbenzoic acid have been determined at 185\u2013190\u2005K. In each crystal, the acid and base mol\u00adecules are linked by a short O\u2014H\u22efN hydrogen bond. 7H4ClNO4\u00b7C10H9N, namely, 2-chloro-4-nitro\u00adbenzoic acid\u20136-methyl\u00adquinoline (1/1), (I), 2-chloro-5-nitro\u00adbenzoic acid\u20136-methyl\u00adquinoline (1/1), (II), 3-chloro-2-nitro\u00adbenzoic acid\u20136-methyl\u00adquinoline (1/1), (III), and 4-chloro-2-nitro\u00adbenzoic acid\u20136-methyl\u00adquinoline (1/1), (IV), have been determined at 185\u2013190\u2005K. In each compound, the acid and base mol\u00adecules are linked by a short hydrogen bond between a carboxyl O atom and an N atom of the base. The O\u22efN distances are 2.5452\u2005(12), 2.6569\u2005(13), 2.5640\u2005(17) and 2.514\u2005(2)\u2005\u00c5, respectively, for compounds (I)\u2013(IV). In the hydrogen-bonded acid\u2013base units of (I), (III) and (IV), the H atoms are each disordered over two positions with O site:N site occupancies of 0.65\u2005(3):0.35\u2005(3), 0.59\u2005(4):0.41\u2005(4) and 0.48\u2005(5):0.52\u2005(5), respectively, for (I), (III) and (IV). The H atom in the hydrogen-bonded unit of (II) is located at the O-atom site. In all of the crystals of (I)\u2013(IV), \u03c0\u2013\u03c0 inter\u00adactions between the quinoline ring system and the benzene ring of the acid mol\u00adecule are observed. In addition, a \u03c0\u2013\u03c0 inter\u00adaction between the benzene rings of adjacent acid mol\u00adecules and a C\u2014H\u22efO hydrogen bond are observed in the crystal of (I), and C\u2014H\u22efO hydrogen bonds and O\u22efCl contacts occur in the crystals of (III) and (IV). These inter\u00admolecular inter\u00adactions connect the acid and base mol\u00adecules, forming a layer structure parallel to the bc plane in (I), a column along the a-axis direction in (II), a layer parallel to the ab plane in (III) and a three-dimensional network in (IV). Hirshfeld surfaces for the title compounds mapped over dnorm and shape index were generated to visualize the weak inter\u00admolecular inter\u00adactions.The structures of the four isomeric compounds of 6-methyl\u00adquinoline with chloro- and nitro-substituted benzoic acids, C Ka values of the acids and bases as well as inter\u00admolecular inter\u00adactions in the crystals. In our ongoing study on crystal structures of the system of quinoline derivatives\u2013chloro- and nitro-substituted benzoic acids, we have shown that three compounds of quinoline with 3-chloro-2-nitro\u00adbenzoic acid, 4-chloro-2-nitro\u00adbenzoic acid and 5-chloro-2-nitro\u00adbenzoic acid, the \u0394pKa [pKa(base)\u00a0\u2212\u00a0pKa(acid)] values of which are 3.08, 2.93 and 3.04, respectively, have a short double-well O\u2014H\u22efN/O\u22efH\u2014N hydrogen bond between the carb\u00adoxy O atom and the aromatic N atom , 8-hy\u00addroxy\u00adquinolinium 3-chloro-2-nitro\u00adbenzoate (\u0394pKa = 3.02) and 3-chloro-2-nitro\u00adbenzoic acid\u20135-nitro\u00adquinoline (1/1) (\u0394pKa = 0.98) , 5-chloro-2-nitro\u00adbenzoic acid\u20135-nitro\u00adquinoline (1/1) (\u0394pKa = 0.94) \u2013(IV).Properties of hydrogen bonds formed between organic acids and organic bases depend on the pThe mol\u00adecular structures of compounds (I)\u2013(IV) are shown in Fig.\u00a01In the hydrogen-bonded acid\u2013base unit of compound (I)Similar to (I)P212121. In the acid\u2013base unit, the quinoline ring system and the benzene ring of the acid are slightly twisted to each other with a dihedral angle of 14.50\u2005(5)\u00b0. The quinoline ring system and the carb\u00adoxy group are also slightly twisted with a dihedral angle of 12.55\u2005(18)\u00b0. The benzene ring makes dihedral angles of 3.14\u2005(18) and 85.04\u2005(11)\u00b0, respectively, with the carb\u00adoxy group and the nitro group.Compound (III)Cc. In the acid\u2013base unit, the quinoline ring system and the benzene ring of the acid are twisted to each other with a dihedral angle of 30.39\u2005(9)\u00b0. The quinoline ring system and the carb\u00adoxy group are also twisted with a dihedral angle of 21.7\u2005(3)\u00b0. The benzene ring makes dihedral angles of 16.4\u2005(3) and 74.4\u2005(3)\u00b0, respectively, with the carb\u00adoxy group and the nitro group.Compound (IV)i; symmetry code as given in Table\u00a01c-axis direction \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01]. The dimeric units are further linked into a column structure stacked along the b-axis direction through a weak \u03c0\u2013\u03c0 inter\u00adaction between the benzene rings with Cg1\u22efCg1iv = 3.9401\u2005(6)\u2005\u00c5 . The mol\u00adecular chains are thus stacked into a layer parallel to the bc plane via these \u03c0\u2013\u03c0 inter\u00adactions.In the crystal of (I)on Fig.\u00a02. The aciit Fig.\u00a03; the cenvia \u03c0\u2013\u03c0 inter\u00adactions between the acid benzene ring and the quinoline ring system, so that the hydrogen-bonded acid\u2013base units related by an inversion center are linked into a column structure along the a-axis direction \u2212x, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01; (ii) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01]. There are no significant inter\u00adactions between the columns.In the crystal of (II)on Fig.\u00a04. The ceni; symmetry code as in Table\u00a03b-axis direction , 3.6126\u2005(8) and 3.6393\u2005(8)\u2005\u00c5, respectively, for Cg1\u22efCg2iii, Cg1\u22efCg3iii and Cg1\u22efCg3iv, where Cg1, Cg2 and Cg3 are the centroids of the C1\u2013C6, N2/C8\u2013C11/C16 and C11\u2013C16 rings, respectively is observed between the layers.In the crystal of (III)on Fig.\u00a05. The aciem Fig.\u00a06, and thuc-axis direction \u2005\u00c5; (ii) x, \u2212y, z\u00a0+\u00a0a-axis direction via \u03c0\u2013\u03c0 inter\u00adactions between the acid ring and the quinoline ring system x\u00a0\u2212\u00a0y\u00a0+\u00a0z\u00a0\u2212\u00a0x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0\u2212\u00a0In the crystal of (IV)on Fig.\u00a07 via C\u2014H\u22efem Fig.\u00a08, and thudnorm and shape index \u2013(IV) mapped over et al., 2016trans-but-2-enedioic acid) \u2013(IV) were obtained by slow evaporation from aceto\u00adnitrile solutions of 6-methyl\u00adquinoline with chloro-nitro\u00adbenzoic acids in a 1:1 molar ratio at room temperature [80\u2005ml aceto\u00adnitrile solution of 6-methyl\u00adquinoline (0.20\u2005g) and chloro-nitro\u00adbenzoic acid (0.28\u2005g for each acid)].Uiso(H) = 1.5Ueq(N or O); the refined distances are given in Tables 1Uiso(H) = 1.2 or 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989020013134/hb7946sup1.cifCrystal structure: contains datablock(s) global, I, II, III, IV. DOI: 10.1107/S2056989020013134/hb7946Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989020013134/hb7946IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989020013134/hb7946IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 10.1107/S2056989020013134/hb7946IVsup5.hklStructure factors: contains datablock(s) IV. DOI: Click here for additional data file.10.1107/S2056989020013134/hb7946Isup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020013134/hb7946IIsup7.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020013134/hb7946IIIsup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020013134/hb7946IVsup9.cmlSupporting information file. DOI: 2034476, 2034475, 2034474, 2034473CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The cobalt core exhibits an approximately square-pyramidal geometry with partially reduced thienyl radical monoanionic ligands. The supra\u00admolecular network is consolidated by hydrogen-bonding inter\u00adactions primarily with nitro\u00adgen, sulfur and chlorine atoms, as well as parallel displaced \u03c0-stacking of the aryl rings. The UV\u2013vis, IR, and CV data are also consistent with monoanionic di\u00adthiol\u00adene ligands and an overall CoII oxidation state.The title compound, bis\u00ad(1,2-diphenyl-2-sulfanyl\u00adidene\u00adethane\u00adthiol\u00adato-\u03ba The cobalt di\u00adthiol\u00adene core is approximately planar, with angles of 89.73\u2005(2)\u00b0 for S1\u2014Co1\u2014S2, 88.93\u2005(2)\u00b0 for S1\u2014Co1\u2014S3, 88.41\u2005(2)\u00b0 for S2\u2014Co1\u2014S4, and 89.90\u2005(2)\u00b0 for S3\u2014Co1\u2014S4. The sum of the angles is 356.97\u2005(4)\u00b0, consistent with only a slight distortion from planarity. The PTA ligand occupies a 5th coordination site with angles of 101.94\u2005(2)\u00b0 for P1\u2014Co1\u2014S1, 98.12\u2005(2)\u00b0 for P1\u2014Co1\u2014S2, 90.97\u2005(2)\u00b0 for P1\u2014Co1\u2014S3, and 97.22\u2005(2)\u00b0 for P1\u2013Co1\u2014S4. Therefore, the overall geometry of [Co(pdt)2(PTA)] is approximately square pyramidal.[Co(pdt)p-anisyl-substituted analogues with PMe3 and PPh3 2(PTA)] is attributed to the small cone angle of 103\u00b0 \u2005\u00c5 for Co1\u2014S1, 2.1669\u2005(6)\u2005\u00c5 for Co1\u2014S2, 2.1685\u2005(5)\u2005\u00c5 for Co1\u2014S3, and 2.1487\u2005(5)\u2005\u00c5 for Co1\u2014S4. These are mostly within the range of Co\u2014S distances of 2.1659\u2005(9)\u20132.1765\u2005(9)\u2005\u00c5 reported for the p-anisyl-substituted analogues with PMe3 and PPh3, respectively \u2005\u00c5 for S1\u2014C7, 1.719\u2005(2)\u2005\u00c5 for S2\u2014C8, 1.729\u2005(2)\u2005\u00c5 for S3\u2014C21, and 1.731\u2005(2)\u2005\u00c5 for S4\u2014C22. These are mostly within the range of S\u2014C distances of 1.721\u2005(2)\u20131.726\u2005(3) and 1.730\u2005(3)\u20131.742\u2005(3)\u2005\u00c5 reported for the 2(PTA)] and one mol\u00adecule of 1,2-dicholorethane are present in the unit cell, as depicted in Fig.\u00a03ii distance of 2.78\u2005\u00c5 \u00c5 Figs. 3, #2 \u25b8 anb axis, parallel displaced \u03c0-stacking of the aryl rings is revealed \u2005\u00c5 apart \u00b0, while the sum of the angles is 356.97\u2005(6)\u00b0 for the PMe3 complex. Similarly, the phosphine in PPh3 is axially distorted because of the steric bulk of the phenyl groups, resulting in two more obtuse bond angles for S2\u2014Co1\u2014P1 and S3\u2014Co1\u2014P1 of 101.31\u2005(3) and 106.6\u2005(3)\u00b0, respectively. The other bond angles of 92.81\u2005(3)\u00b0 for S1\u2014Co1\u2013P1 and 97.13\u2005(3)\u00b0 for S4\u2014Co1\u2014P1 are within the range of S\u2014Co1\u2014P1 angles of 91.19\u2005(3) to 99.65\u2005(3)\u00b0 observed for the PMe3 complex.A survey of the Cambridge Structural Database ] was conducted in di\u00adchloro\u00admethane transition, based on comparison with the related iron complex with PPh3 complex. The IR signal for [Co(pdt)2(PTA)] at 1157.61\u2005cm\u22121 is characteristic of monoanionic di\u00adthiol\u00adenes with a \u03c0-radical when coordinated to metals, and is attributed to \u03bd(C=S\u00b7) ne Fig.\u00a05 and reve2(PTA)] was collected in a solution of dichoro\u00admethane with a platinum working electrode of [Co(pdt)de Fig.\u00a06 and a glde Fig.\u00a06. Both CV2(pdt)4] and PTA under an N2 atmosphere. To this mixture of solids, 20\u2005mL of CH2Cl2 were added and stirred for 4\u2005h at room temperature. The solvent was removed under reduced pressure and the resulting dark-orange solid was washed with 3 \u00d7 5\u2005mL of Et2O and dried under vacuum. The product was stable under reduced pressure and at room temperature. Yield: 92% . Crystals suitable for X-ray diffraction were grown by the vapor diffusion method with diffusion of pentane over a 1,2-di\u00adchloro\u00adethane solution of the compound. UV\u2013Vis spectra were obtained at ambient temperature with a Varian Cary 50 diode array spectrometer, while IR spectra were taken neat with an ALPHA FTIR instrument. Electrochemical measurements were performed with a CHI600E electroanalyzer workstation using an Ag/AgCl reference electrode, a platinum disk working electrode, a platinum wire auxiliary electrode, and [nBu4N][PF6] as the supporting electrolyte in CH2Cl2. Under these conditions, the [Cp2Fe]+/ Cp2Fe couple consistently occurred at +440\u2005mV. UV\u2013vis in CH2Cl2: : 301\u2005nm (17881), 877\u2005nm (6428). IR spectroscopy (cm\u22121): 3366.53 (w), 3054.34 (w), 2928.02 (w), 2869.68 (w), 1592.50 (w), 1440.63 (m), 1415.07 (s), 1275.25 (m), 1240.54 (w), 1157.61 (m), 1091.56 (s), 1012.49 (m), 969.10 (s), 940.65 (vs), 739.50 (s), 693.31 (s).A 50\u2005mL Schlenk flask containing a stir bar was charged with [Co2, respectively, and refined using a riding model with Uiso(H) = 1.2 Ueq(C) for CH and CH2.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989020005447/zl2776sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020005447/zl2776Isup2.hklStructure factors: contains datablock(s) I. DOI: 1986931CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-019-42476-4, published online 11 April 2019Correction to: The original version of this Article contained errors.In the Abstract,\u201cThe comparison of the mean load to failure of all 3 groups did not reveal a significant difference.\u201dnow reads:\u201cThe comparison of the mean load to failure of all 3 groups did not reveal a significant difference.\u201dIn Table 1,2\u201d.\u201cStiffness in N/mmnow reads:\u201cStiffness in N/mm\u201d2\u201d, row r7,Furthermore, in column \u201cStrength in N/mm\u201c-27.1\u201dnow reads:\u201c27.1\u201dAdditionally Table 1 contained errors in the values in columns Stiffness in N/mm, Strength in N/mm\u00b2, Cross sectional area in mm\u00b2. The original Table\u00a0As a result of this, in the Results section,2 . There was no significant difference between mean stiffness and strength . A detailed overview is presented in Table\u00a0\u201cThe mean cross sectional area of ROI (fracture region) was 1.81 mmnow reads:2 .There was no significant difference between mean stiffness and strength . A detailed overview is presented in Table\u00a0\u201cThe mean cross sectional area of ROI (fracture region) was 1.72 mmAnd,\u201cNo significant difference in the load to failure values between the three groups was found (p\u2009\u2009=\u2009\u20090.113). The mean load to failure in group 1 was 28.7\u2009\u2009\u00b1\u2009\u20096.1\u2009N compared to 23.8\u2009\u2009\u00b1\u2009\u20093.8\u2009N in group 2. The mean load to failure in group 3 was 23.7\u2009\u2009\u00b1\u2009\u20095.7\u2009N .\u201dnow reads:\u201cNo significant difference in the load to failure values between the three groups was found (p\u2009=\u20090.113). The mean load to failure in group 1 was 28.7\u2009\u00b1\u20096.4\u2009N compared to 23.7\u2009\u00b1\u20096.0\u2009N in group 2. The mean load to failure in group 3 was 24.0\u2009\u00b1\u20093.9\u2009N .\u201dThese errors have now been corrected in the PDF and HTML versions of the Article."} +{"text": "Hydrogen bonding of the anions is restricted to F-atom acceptors only, with particularly strong N\u2013H\u22efF inter\u00adactions [N\u22efF = 2.5072\u2005(15)\u2005\u00c5] established by axial and cis-positioned equatorial F atoms.In the structure of the title salt, second-order Jahn\u2013Teller distortion of the coordination octa\u00adhedra around V ions is reflected by coexistence of short V\u2014O bonds and 7H12N6)[VOF5], second-order Jahn\u2013Teller distortion of the coordination octa\u00adhedra around V ions is reflected by coexistence of short V\u2014O bonds [1.5767\u2005(12)\u2005\u00c5] and trans-positioned long V\u2014F bonds [2.0981\u2005(9)\u2005\u00c5], with four equatorial V\u2014F distances being inter\u00admediate in magnitude [1.7977\u2005(9)\u20131.8913\u2005(9)\u2005\u00c5]. Hydrogen bonding of the anions is restricted to F-atom acceptors only, with particularly strong N\u2013H\u22efF inter\u00adactions [N\u22efF = 2.5072\u2005(15)\u2005\u00c5] established by axial and cis-positioned equatorial F atoms. Hirshfeld surface analysis indicates that the most important inter\u00adactions are overwhelmingly H\u22efF/F\u22efH, accounting for 74.4 and 36.8% of the contacts for the individual anions and cations, respectively. Weak CH\u22efF and CH\u22efN bonds are essential for generation of three-dimensional structure.In the structure of the title salt, (C MvOF5]2\u2212 series of ions [VOF5] [VOF5] \u00b7[VOF5]2\u2212, which gives insight into the hydrogen-bonding behaviour of [VOF5]2\u2212 anions when combined with the bitopic nitro\u00adgen-rich 4,4\u2032-bis dication. This cation provides different kinds of hydrogen-bond donor sites complemented by triazole-N acceptors, which are relevant to many types of coordination and hydrogen-bonded systems \u2005\u00c5], which define the local polar axis of the anion. Four equatorial V\u2014F bonds 2\u2212 anions in the salts with (H2bipy)2+ 2+ cations = 103.46\u2005(12) and 104.11\u2005(12)\u00b0 \u00b0 and N3\u2014C5\u2014C6\u2014C7 of \u221263.73\u2005(17)\u00b0. A diversity of metal complexes suggest nearlys equal occurrence of trans\u2013gauche and all-trans sequences for the present moiety \u2005\u00c5 .Primary strong N\u2014H\u22efF bonding links the ionic counterparts into chains, which aggregate forming layers parallel to the ks Fig.\u00a02. The firThe packing of the layers extends the structure in the third dimension. For every next layer of the succession, the direction of the primary N\u2014H\u22efF bonded chains is inclined by 56.8\u00b0 to the direction of chains from the preceding layer Fig.\u00a03. Links bet al., 2004CrystalExplorer17 to 1.3445 (blue) a.u. indicates a number of red spots related to hydrogen-bond contacts. Particularly prominent spots are associated with the strongest N\u2014H\u22efF bonds. However, even the C\u2014H\u22efF inter\u00adaction with the weakest of the present donors is reflected by a red spot on the surface . In addition to this very sharp spike, the plot clearly reveals the more subtle feature of anion\u22ef\u03c0 bonding, which appears as a short spike at 2.7\u2005\u00c5 , 0.84\u2005ml of 7% aqueous HF solution (3.0\u2005mmol) and 2\u2005ml of water were placed in a Teflon vessel and heated in a steel bomb at 413\u2005K for 24\u2005h. Cooling to room temperature over a period of 48\u2005h afforded colourless crystals of the title salt, in a yield of 27\u2005mg (40%). Analysis (%) calculated for C7H12F5N6OV: C 24.57, H 3.54, N 24.57; found: C 24.38, H 3.49, N 24.70.The bitriazole was prepared in a yield of 33% by the acid-catalysed condensation of 1,3-di\u00adamino\u00adpropane and 2) = 0.98\u2005\u00c5; Uiso(H) = 1.2Ueq(CH) and 1.5Ueq(NH).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698902000585X/hb7908sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S205698902000585X/hb7908Isup2.hklStructure factors: contains datablock(s) I. DOI: 1999654CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Of these compounds, 10 and 12 form 1D or 2D coordination polymers in the solid state. (Spectro)Electrochemical studies confirmed reversible Fc/Fc+ redox events between \u2212130 and 160\u2005mV. 1,6\u2010 and 2,7\u2010Substitution in 5\u2009a (E\u00b0\u2032=\u2212130\u2005mV) and 7\u2009a (E\u00b0\u2032=50\u2005mV) influences the redox potentials, whereas the ones of 5\u2009b and 7\u2009b (E\u00b0\u2032=20\u2005mV) are independent. Compounds\u20055\u2009b, 7\u2009b, 10, and 12 show single Fc oxidation processes with redox splittings between 70 and 100\u2005mV. UV/Vis/NIR spectroelectrochemistry confirmed a weak electron transfer between FeII/FeIII in mixed\u2010valent [5\u2009b]+ and [12]+. DFT calculations showed that 5\u2009bnon\u2010covalently interacts with the single\u2010walled carbon nanotube (SWCNT) sidewalls as proven by, for example, disentangling experiments. In addition, CV studies of the as\u2010obtained dispersions confirmed exohedral attachment of 5\u2009b at the SWCNTs.The synthesis of 1\u2010Fc\u2010 (\u03b75\u2010C5H5)) is discussed. Two of these compounds form 1D or 2D coordination polymers in the solid state. (Spectro)electrochemical studies confirmed reversible Fc/Fc+ redox events between \u2212130 and 160\u2005mV. With the example of 2,5\u2010Fc2\u2010cC4H2E (Fc=Fe(\u03b75\u2010C5H4)(\u03b75\u2010C5H5); E=O, S, NR), it could be shown that similar geometries and hence comparable electrostatic interactions enable a correlation between electrochemical and spectroscopic properties.Redox\u2010active ferrocenyl\u2010functionalized five\u2010non\u2010covalent interactions, for example, \u03c0\u2013\u03c0 stacking.Furthermore, aromatics are suitable to modify carbon nanotubes (CNTs) by + (NADH=dihydronicotinamide adenine dinucleotide).Ferrocene\u2010functionalized CNT nanohybrids have attracted much attention in the fields of (bio)analytical electrochemistry,N\u2010(pyren\u20101\u2010ylmethyl)propanamide to SWCNTs and tested as glucose sensor electrode materials.6)2.II/RuIII redox potential is stable over 200 cycles. Another hybrid material is CNT/cobalt bis\u2010pentyl]butyramide).Non\u2010covalently functionalized CNTs bearing a ferrocenyl substituent were recently obtained through \u03c0\u2010stacking of 3\u2010ferrocenyl\u2010Lately, we became interested in attaching redox\u2010active Fc\u2010functionalized polyaromatic hydrocarbons to SWCNTs as a way to achieve debundeling as well as to influence the electronic properties of the thus\u2010modified carbon nanotubes and to use them as field\u2010effect sensors.2\u2010, and 2,7\u2010Fc2\u2010pyrenes, 3,6\u2010Fc2\u20109,10\u2010phenanthrenedione, and 3,6\u2010Fc2\u20109,10\u2010dimethoxyphenanthrene compounds is discussed. Their (spectro)electrochemical properties and their molecular structures in the solid state are reported. DFT calculations for 1,6\u2010Fc2\u2010pyrene and semiconducting and metallic SWCNTs are provided, as well as the use of 1,6\u2010Fc2\u2010pyrene to disentangle SWCNTs. The electrochemical properties of the as\u2010obtained dispersions are presented.Hence, within this study, the synthesis and characterization of a series of 1\u2010Fc\u2010, 1,6\u2010Fc3, 5\u2009a,b, 7\u2009a, 10) or Negishi C\u2212C cross\u2010coupling reactions were applied (\u03b75\u2010C5H5)) was treated with the respective 1\u2010Br\u2010 (2), 1,6\u2010Br2\u2010 (4), 2,7\u2010Br2\u2010pyrene (6), or 3,6\u2010Br2\u20109,10\u2010phenanthrenedione (9) in the presence of [PdCl2(dppf)] ferrocene) as the catalyst in the molar ratio of 1:1 or 2:1 in boiling toluene. After appropriate work\u2010up, compounds\u20053, 5\u2009a,b, 7\u2009a, and 10 could be isolated as orange or green (10) solids in a yield of 3\u201365\u2009% . For comparison, compound 9\u2010ferrocenylphenanthren was synthesized according to the Suzuki C\u2212C cross\u2010coupling protocol used within the synthesis of 3, 5\u2009a,b, 7\u2009a, and 10 .Ferrocene boronic acid FcB(OH)1 with 1,6\u2010Br2\u2010C16H8 (4) in the molar ratio of 2:1, a mixture of the respective mono\u2010 and diferrocenyl\u2010functionalized pyrenes\u20055\u2009a,b was formed .In the transmetalation reaction of 1 with 2,7\u2010Br2\u2010C16H8 (6) in a 2:1 ratio in boiling toluene solely gave the monoferrocenyl\u2010substituted species 2\u2010Br\u20107\u2010Fc\u2010pyrene (7\u2009a), which was obtained as an orange solid in a yield of 32\u2009% after column chromatography 6 in the presence of 1.0\u2005mol\u2009% [PdCl2(dppf)] as the catalyst in boiling tetrahydrofuran produced 7\u2009b, however, in a yield as low as 2\u2009% . In this reaction, other than 7\u2009b, ferrocene was formed as the main product. Compound\u20057\u2009b is a very poorly soluble species in common polar organic solvents. Once 7\u2009b was isolated in its solid form, it was almost impossible to redissolve it and hence spectroscopic and electrochemical experimental data on this organometallic compound are limited.However, diferrocenyl pyrene\u200510 and 12 were synthesized by either the Suzuki\u2013Miyaura (10) or the Negishi (12) reaction starting from 3,6\u2010dibromophenanthrene\u20109,10\u2010dione 9 in toluene in the presence of catalytic amounts of [PdCl2(dppf)] led to the formation of green 3,6\u2010diferrocenylphenanthrene\u20109,10\u2010dione (10) in a yield of 28\u2009%. However, the synthetic methodology described by Phulwale et\u2005al. for the reduction of the ketone functionalities in 10 did not lead to the formation of the respective 3,6\u2010diferrocenyl\u20109,10\u2010dimethoxyphenanthrene compound\u200512.9 was performed prior to the C\u2212C cross\u2010coupling reaction. Thus, compound\u200511 could be isolated in virtual quantitative yield. Upon treatment of 11 with two equivalents of FcZnCl (8) in tetrahydrofuran as solvent for 24\u2005h in the presence of [PdCl2(dppf)] gave orange 12 in an overall yield of 13\u2009% .Compounds\u20055\u2009a,b, 7\u2009a,b, 10, and 12 was confirmed by elemental analysis, IR, 1H and 13C{1H} NMR spectroscopy, and high\u2010resolution ESI\u2010TOF mass\u2010spectrometry , whereas the physical and analytical data of 3 are documented in reference\u20057\u2009b).The identity of 1H, 13C{1H} NMR) data of 3, 5\u2009a,b, 7\u2009a,b, 10, and 12 are consistent with their formulations as mono\u2010 or diferrocenyl\u2010functionalized pyrene, 9,10\u2010phenanthrenedione, and 9,10\u2010dimethoxyphenanthrene compounds, showing the characteristic coupling patterns of the appropriate isomers .7\u2009b no reliable 13C{1H} NMR data could be obtained, owing to its low solubility in common polar organic solvents (see above).The spectroscopic , I2/a (5\u2009a), P21/n (5\u2009b), or P21/c and in the orthorhombic space groups Pnn2 (12) and P212121 (9\u2010ferrocenylphenanthrene) with one or two (10) independent molecules in the asymmetric unit. Intermolecular \u03c0\u2013\u03c0 interactions were investigated between the aromatic entities and the ferrocenyl cyclopentadienyl units .The compounds crystallize in the monoclinic space groups ipso\u2010carbon atoms and the aromatic units is similar for all investigated compounds (from 1.476(3)\u2005\u00c5 (7\u2009a) to 1.554(8)\u2005\u00c5 (10)) and agree well with literature values.The distance between the ferrocenyl cyclopentadienyl to 1.554\u2005\u00c5 (10)) In Tables\u2005SI3 and SI4 (in the Supporting Information), the bond lengths of the different aromatic cores are summarized. The substitution pattern does not affect the position of the single and double bonds in the aromatic moieties. This observation is in good agreement with values published for other substituted pyrenyl and phenanthrenyl derivatives.3, 5\u2009a,b, 7\u2009a,b, 12, and 9\u2010ferrocenylphenanthrene. Compounds containing hydrogen atoms in position 8 with regard to the Fc group possess increased values of 31.5(5)\u00b0 (5\u2009b) to 40.05(10)\u00b0 (5\u2009a), owing to a steric interaction with the \u03b1\u2010H atoms of the ferrocenyls. In contrast, the geometric properties of the 2\u2010 or 3\u2010substitution pattern allow for more planar intersections between 0.1(3)\u00b0 (10) and 22.1(9)\u00b0 (12). Hence, these compounds should be preferred to achieve a high degree of electron transfer interaction between the \u03c0\u2010system of the ferrocenyls and the aromatic cores.3, 5\u2009a,b, and 9\u2010ferrocenylphenanthrene argue for a weak interaction. In addition, the rms distortions (root mean square deviations) of the condensed aromatic units were calculated and are summarized in Table\u2005SI5 (in the Supporting Information), where 5\u2009a (0.0497) and 12 (0.0680) showed exceptional high values.The rotation of the ferrocenyls out of the aromatic co\u2010planarity was examined for 5\u2009b, 7\u2009b, 10, and 12 exhibit an anti\u2010positioning of the Fc groups towards each other with regard to the aromatic planes.10) to 12.7288(8)\u2005\u00c5 (7\u2009b). Thereby, the Fe\u22c5\u22c5\u22c5Fe distances in 10 (9.0374(13)\u2005\u00c5 Fe1\u2013Fe2; 9.0390(13)\u2005\u00c5, Fe3\u2013Fe4) and 12 (10.123(6)\u2005\u00c5) point towards a better electronic coupling between the ferrocenyl groups than in 5\u2009b (11.477(2)\u2005\u00c5) and 7\u2009b (12.7288(8)\u2005\u00c5).The diferrocenyl\u2010substituted compounds\u20053, 5\u2009a,b, 7\u2009a,b, 10, 12, and 9\u2010ferrocenylphenanthrene as \u03c0\u2010surfactants on CNTs, the interaction through intermolecular \u03c0/C\u2212H\u22c5\u22c5\u22c5\u03c0 bonding was investigated for the aromatic and ferrocenyl units . Except for 7\u2009b, all compounds show intermolecular \u03c0\u2013\u03c0 interactions , of which 3, 5\u2009b, and 9\u2010ferrocenylphenanthrene show T\u2010shaped \u03c0\u2010interactions between C5H5/C5H4 units and the aromatic moieties with distances of 4.5276(14) (3)\u20134.994(6)\u2005\u00c5 (5\u2009b). The solid\u2010state structures of 5\u2009a and 7\u2009a show T\u2010shaped and parallel displaced \u03c0\u2010interactions with distances between 4.6081(17) (5\u2009a) and 4.6925(14)\u2005\u00c5 (7\u2009a) and angles from 84.68(15) (5\u2009a) to 89.24(14)\u00b0 (7\u2009a). The parallel displaced \u03c0\u2010interactions between the pyrene cores of 5\u2009a and 7\u2009a show distances of approximately 3.6\u2005\u00c5, forming dimers .With regard to the utilization of 10 and 12, the parallel displaced \u03c0\u2010interactions between the arenes led to the formation of 1D (12) and 2D coordination polymers (10) along the a\u2010 (10) or b\u2010axis (12), resulting in the formation of columns . The columnar structure of 12 is based on the parallel displaced \u03c0\u2010interactions involving C11\u2013C16 of the phenanthrene with centroid\u2013centroid distances of 5.165(12)\u2005\u00c5, exceeding the criterion for \u03c0\u2010stacking (3.3\u20133.8\u2005\u00c5).In case of 10 builds a 2D coordination polymer, whereby both molecules of the asymmetric unit are responsible for the columnar structure. Parallel displaced \u03c0\u2010interactions between the ferrocenyl's cyclopentadienyls interconnect the columns. The arrangement along the a\u2010axis is based on the interactions between the arenes of the phenanthrene backbone with distances ranging from 3.589(3) to 4.549(3)\u2005\u00c5. Furthermore, this stacking is supported by \u03c0\u2010interactions involving the Fc\u2010C5H4 units and the aromatic moieties with distances ranging from 3.466(3) to 4.800(3)\u2005\u00c5 .Compound\u2005\u1e7dmax) of compound\u200510 were measured in a set of 23 common organic solvents of different polarities and hydrogen\u2010bonding abilities =658\u2005nm =553\u2005nm). This shift corresponds to a positive solvatochromism with a solvatochromic range of \u0394\u1e7dmax (10)=2880\u2005cm\u22121.Compound\u2005The interactions of solvatochromic dyes with pure solvents or solvent mixtures arise from a combination of many effects.\u03b1, \u03b2, and \u03c0*. In addition, \u1e7dmax,0 corresponds to a standard process, referenced to a nonpolar medium. The parameters a, b, and s represent solvent\u2010independent correlation coefficients, which reflect the relative effect of each of the three parameter \u03b1, \u03b2, and \u03c0*.According to Equation\u2005(1), the effect of hydrogen\u2010bonding donor capacity (HBD),\u03b1, \u03b2, and \u03c0* used for the multiple linear regression analysis are given in Table\u2005SI6 (in the Supporting Information).10, which is qualitatively the best according to the solvent scales of Kamlet\u2013Taft, is given in Table\u2005The solvent parameters r is greater than 0.90 for the LSE relationship, indicating a high validity of the multiparameter equation, allowing significant conclusions to be drawn. Compound\u200510 shows a positive solvatochromism with increasing acidity and dipolarity/polarizability of the solvents.The correlation coefficient s indicates that the electronically excited state of 10 becomes solvated stronger and is consequently stabilized with increasing solvent dipolarity/polarizability. Owing to the strength of the higher dipole moment, the energy of the electronically excited state decreases more than the energy of the ground state. This is in good agreement with a bathochromic shift of the UV/Vis absorption maxima with increased polarity of the solvent.The negative sign of a for 10 indicates that there is a bathochromic shift of \u03bbmax with increasing hydrogen\u2010bond donor capacity of the solvent (positive solvatochromism). Owing to the solvation of the carbonyl oxygen atom by HBD solvents the push\u2013pull character becomes enhanced. This effect is on the same order of magnitude as the influence of the dipolarity/polarizability. The hydrogen\u2010bond acceptor capacity of the solvents, on the other hand, has no influence on the solvatochromic behavior of 10 (b=0).The negative sign of parameter \u1e7dmax) of 1820\u20132460\u2005cm\u22121 in contrast to 10 with 2880\u2005cm\u22121, which indicates a stronger solvatochromic behavior.A similar behavior was observed for ferrocenyl\u2010substituted maleimides.3, 5\u2009a,b, 7\u2009a,b, 10, and 12 were investigated by cyclic voltammetry (CV), square wave voltammetry (SWV), and spectroelectrochemistry (in situ UV/Vis/NIR). As supporting electrolyte, an anhydrous dichloromethane solution containing 0.1\u2005mol\u2009L\u22121 of [NnBu4][B(C6F5)4] was used.6]\u2212 or [Cl]\u2212, [B(C6F5)4]\u2212 stabilizes highly charged species in solution and minimizes ion pairing effects.+ redox couple.\u22121 are summarized in Table\u2005The redox properties of compounds\u20053 as well as 5\u2009b, 7\u2009b, 10, and 12 show one or two reversible redox processes. The aromatic cores are characterized by a reversible (5\u2009a) or irreversible redox event between 950 and 1250\u2005mV, however, for 7\u2009b, 10, and 9\u2010ferrocenylphenanthrene no oxidation of the aromatic core occurred under the applied measurement conditions.The Fc groups of 9\u2010ferrocenylphenanthrene and 5\u2009a, 7\u2009a), the substitution pattern and the nature of the aromatic core . Comparison with 3 shows that the bromo substituent has a strong impact on the electron density at Fc, shifting the potential by 160\u2005mV anodically. The Fc\u2010based redox process of 7\u2009a is further shifted to 50\u2005mV (vs. FcH/FcH+) compared with 3. Positions 2 and 7 are electron\u2010poor5\u2009a. This observation is in contrast to the investigations of Fc\u2010substituted naphthalenes, where the influence of the substitution pattern is stronger than the impact of electron\u2010withdrawing entities like bromine.3 and 9\u2010ferrocenylphenanthrene, respectively. Dhokale et\u2005al. reported that 1\u2010ferrocenylpyrene and 1\u2010ferrocenylethynylpyrene show Fc\u2010based oxidations at 50 and 70\u2005mV when using [NnBu4][PF6] as the electrolyte versus a saturated calomel electrode (SCE).nBu4][B(C6F5)4] as a weakly coordinating electrolyte.Within the series of monoferrocenyl\u2010substituted pyrenes, compound\u20055\u2009b and 7\u2009b show a single redox event for both ferrocenyl groups at 20\u2005mV and 100\u2005mV pyrene by using [NnBu4][PF6] as the electrolyte.tert\u2010butyl\u20109\u2009H\u2010carbazol\u20109\u2010yl)pyrene) to 100\u2005mV pyrene), which is in good agreement with the results observed for 5\u2009b and 7\u2009b.5\u2010C5Me5)(dppe)RuC\u2261C)2\u2010pyrene, which showed a value of 120\u2005mV.Diferrocenyl pyrenes es Table\u2005 point to10 and 12 were synthesized to investigate the influence of electron\u2010withdrawing and \u2010donating groups on the redox potential of the ferrocenyl oxidation. The methoxy\u2010functionalized compound\u200512 shows a redox process at 30\u2005mV (vs. FcH/FcH+). In contrast to that, the electron\u2010withdrawing keto groups in positions 9 and 10 of 10 decrease the electron density of the Fc entities and therefore shift the respective potential by 130\u2005mV anodically. The redox separation of 10 and 12 is lower than for the appropriate diferrocenyl\u2010substituted pyrenes\u20055\u2009b and 7\u2009b. By deconvolution of the SW voltammograms, redox splittings of 70 (10) and 80\u2005mV (12) were found, pointing to a weak metal\u2013metal interaction based on either electrostatic interactions or electron transfer measurements were performed. Compound\u20057\u2009b could not be investigated by this method, owing to its low solubility. Furthermore, the UV/Vis/NIR investigations of 10 revealed no intervalence charge transfer (IVCT) absorption but a MLCT transition, which is in good agreement with the postulated resonance structures (Scheme\u2005SI2 in the Supporting Information).For the purpose of classification, the electron transfer between the Fc/Fc5\u2009b and 12) or propylene carbonate solutions (12) containing 5\u2009b or 12 (2.0\u2005mmol\u2009L\u22121) and [NnBu4][B(C6F5)4] (0.1\u2005mol\u2009L\u22121) as supporting electrolyte in an OTTLE cell.5\u2009b and 12. During the measurements, oxidation of the neutral compounds to mixed\u2010valent [5\u2009b]+ and [12]+ and finally to the fully oxidized species [5\u2009b]2+ and [12]2+ was observed gave mixed\u2010valent species [5\u2009b]+ and [12]+ and hence a broad absorption within this area (800\u20133000\u2005nm) was observed. A further increase in the potential resulted in a decrease of this band, which is characteristic for intervalence charge transfer excitations and therefore the small comproportionation constants lead, in the best case, to a mixture consisting of an equilibrium of 73\u2009% mixed\u2010valent monocationic [12]+ as well as 13.5\u2009% neutral 12 and di\u2010cationic species [12]2+ each. As the composition of this equilibrium is not determinable by using an OTTLE cell, the extinction values are associated with considerable uncertainties.For neutral 5\u2009b and 12. The NIR data of the IVCT absorptions are summarized in Table\u2005The observed spectra were deconvoluted into three Gaussian\u2010shaped bands, which represent a LF (ligand field),5\u2009b and 12 showed no absorption in the NIR region, whereas mixed\u2010valent [5\u2009b]+ and [12]+ have weak and broad absorptions between 750 and 2000\u2005nm. The IVCT bands of Fc\u2010substituted aromatics 5\u2009b and 12 exhibit different characteristics . The highest intensity of 510\u2005L\u2009mol\u22121\u2009cm\u22121 was found for 575\u2005mV (vs. Ag/Ag+). From Figure\u2005SI20 (in the Supporting Information), it can be seen that in the series of increasing potential the IVCT absorption increases until its maximum at 575\u2005mV, followed by a decrease of intensity upon further oxidation to [5\u2009b]2+ and an increase of LMCT. The determined values of the IVCT absorption of 5\u2009b are in good agreement with 2,7\u2010bispyreneNeutral cs Table\u2005. For 5\u2009b12 leads to a more intense and narrow IVCT absorption than in Fc2\u2010substituted pyrene 5\u2009b and is further shifted hypsochromically compared with the metal\u2013metal interaction band of 5\u2009b. This statement is based on the higher electron density of the dimethoxy\u2010substituted phenanthrene moiety, which was observed in the CV measurements (Table\u200512 is oxidized more easily (1110\u2005mV vs. FcH/FcH+) than the pyrenyl connectivity of 5\u2009b .The smaller \u03c0\u2010conjugated bridge present in \u03bc=4.9\u2005D) was further chosen as an additional solvent for the spectroelectrochemical measurements of 12. Analogous investigations on 5\u2009b failed in the UV/Vis/NIR setup, owing to its insufficient solubility in propylene carbonate. In the case of [12]+, the IVCT transition could be detected at approximately 11\u2009000\u2005cm\u22121 by using the solvent with higher polarity, revealing a solvatochromic shift (\u0394\u03bdmax) of 1000\u2005cm\u22121 (Table\u2005\u03bdmax in comparison for very strongly coupled systems (\u0394\u03bdmax\u2248100\u2005cm\u22121)Within these species, the IVCT bands typically show strong solvatochromism.3, 5\u2009a,b, 7\u2009a, and 12 were investigated, except for 10, which showed a MLCT . The UV/Vis/NIR spectra are depicted in Figures\u2005SI17\u2013SI19, SI21, SI22, SI24, and SI25 in the Supporting Information). The respective LMCT data of [3]+, [5\u2009a]+, [5\u2009b]2+, [7\u2009a]+, and [12]2+ are summarized in Table\u2005To gain a deeper insight into the communication between the aromatic core and the ferrocenyl ligands, the ligand\u2010to\u2010metal charge transfer properties of E\u00b0\u2032, the transition energy \u03bdmax is increased. The only exception is 1\u2010ferrocenylpyrene\u20053, which shows an anodic shift of its redox event and its transition at 8150\u2005cm\u22121. This might be based on the fact that the plane intersection of 3 in the solid state exhibits a rms value of 0.0348, which indicates a bent aromatic core. Further, the carbon atom C11 shows the highest displacement with 0.0685(17)\u2005\u00c5, confirming a weak interaction between the Fc and the arene moieties (Table\u2005SI5 in the Supporting Information). Equivalent correlations have been reported for charge transfer complexes, for example, ferrocenyl naphthalenes,5\u2009a with 7\u2009a, it is clear that a better communication between the Fc and the pyrene building blocks is given by the substitution in positions 1 and 6 than in 2 and 7, owing to higher intensity of the transition for 5\u2009a.The energy of the LMCT absorptions is related to the redox potentials of the Fc entities, displaying that with an increase of th 0.0685\u2005\u00c5, confi5\u2009b for disentangeling and modification studies of the SWCNTs surface. Hence, DFT calculations were additionally carried out to support the respective pyrene\u2013SWCNT interactions.Based on the spectroelectrochemical properties and that the pyrene connectivity is favored to non\u2010covalently bind to CNTs,5\u2009b and SWCNTs. Here, we have considered both semiconducting \u2010SWCNT and metallic \u2010SWCNT as representative models of SWCNTs. Figure\u20055\u2009b on \u2010SWCNT. It was found that the pyrene group of 5\u2009b tends to orient parallel to the CNT surface owing to optimized \u03c0\u2013\u03c0 interactions. The computed pyrene\u2013CNT equilibrium distance and adsorption energy are 3.2\u2005\u00c5 and \u22121.26\u2005eV by using the PBE (Perdew\u2013Burke\u2013Ernzerhof) functional with empirical dispersion corrections (PBE\u2010D). In contrast, the plain DFT\u2010PBE calculations predict a much larger pyrene\u2013CNT equilibrium distance (4.1\u2005\u00c5) and a negligible adsorption energy (\u22120.1\u2005eV). These data indicate that the van der Waals forces are the dominant interactions between 5\u2009b and SWCNTs. This conclusion was also observed for the adsorption of 5\u2009b on a \u2010SWCNT surface, suggesting that the diameter and chirality of CNTs have only a small influence on the adsorption mechanism. Additionally, Mulliken population analysis was employed to study the charge transfer in the ground state. The calculation results reveal that only a few electrons (about 0.1\u2009e) are transferred from 5\u2009b to the CNT, which further confirms their \u03c0\u2013\u03c0 interactions.Density functional theory (DFT) calculations were performed to provide mechanistic insights into the interactions between 5\u2009b to interact with SWCNTs in a non\u2010covalent fashion in solution, debundeling experiments were performed with two commercially available materials: (i)\u2005chirality\u2010enriched \u2010SWCNTs as powder and (ii)\u2005NanoIntegris IsoSol S\u2010100\u00ae SWCNT in dispersion. \u2010SWCNTs are, owing to their low purity, not suitable for electronic components but are an appropriate model system for the disentangling. However, the NanoIntegris IsoSol S\u2010100\u00ae SWCNT dispersion, consisting of up to 99.9\u2009% semiconducting material, has proven to be an appropriate material for the scalable fabrication of CNT transistors and sensorsTo investigate the ability of 5\u2009b was used for disentangling experiments with the NanoIntegris IsoSol S\u2010100\u00ae SWCNT dispersion and the results thereof are summarized in Figure\u20055\u2009b are presented in Figure\u2005SI26 (in the Supporting Information).As indicated above, compound\u20055\u2009b by using a vacuum filtration procedure aryl\u2010based polymer)Gen1 solid, both the S22 and S33 transitions of the SWCNT mixture as well as the PFH\u2010R transition remain unchanged from a spectral point of view, the relative composition of the absorption spectrum allows us to conclude a first successful depletion step for the reference solid through the vacuum filtration procedure. For the Gen2 solid, the spectrum clearly mimics the SWCNT signature from the Gen0 and Gen1 spectra as well as the particular PFH\u2010R transition (spectrum PFH\u2010R (toluene) mathematically deconvoluted from the spill liquid and the Gen0 spectrum). Additionally, the spectrum for the Gen2 solid shows a transition at 352\u2005nm, which can be identified as an absorption feature of 5\u2009b (measured spectrum 5\u2009b (toluene)), which unambiguously indicates that 5\u2009b is present in the redispersed SWCNT solid after the vacuum filtration procedure.In spectrum 0 Figure\u2005, the S225\u2009b was further confirmed by electrochemical measurements. The redox properties of the dispersion Gen2 ({5\u2009b}{PFH\u2010R}@SWCNT) were investigated by cyclic voltammetry by using Gen2\u2010modified graphene paper\u22121 KCl. The voltammetry measurements were performed at 25\u2009\u00b0C. All potentials are referenced to the SCE using K3[Fe(CN)6] as the internal standard.The functionalization of SWCNTs with Gen2 show a ferrocenyl\u2010based redox process at 380\u2005mV .The CVs of Ep value of the respective redox processes are characteristic.\u22121; Table\u2005i\u2248vr0.62; vr=sweep rate), which is far from a linear relationship, but also does not reflect the i\u2248\u221avr of a fully dissolved redox system. The redox processes of dispersion Gen2 show \u0394Ep values between 40 and 55\u2005mV . This emphasizes that 5\u2009b is exohedrally attached to the SWCNTs. In Gen2, where 5\u2009b is \u03c0\u2010bonded to SWCNTs, but not immobilized at the electrode surface, the above criteria for surface\u2010confined redox couples are unfulfilled, whereas simultaneously the electrochemical system Gen2 does not behave accordingly to homogeneously dissolved redox\u2010active molecules.To exclude that the redox process of + event is quite stable after polymerization, as the peak current decreases only slightly after five cycles .To investigate the influence of the pyrene core towards the ferrocenyl\u2010based redox process, the potential area was extended to 900\u2005mV Figure\u2005. An irre3), 1\u2010Br\u20106\u2010Fc\u2010 (5\u2009a), 2\u2010Br\u20107\u2010Fc\u2010 (7\u2009a), 1,6\u2010Fc2\u2010pyrene (5\u2009b), and 3,6\u2010Fc2\u20109,10\u2010phenanthrenedione (10) were synthesized by reacting ferrocene boronic acid (1) with the respective bromo\u2010functionalized aromatics by applying the Pd\u2010catalyzed Suzuki C\u2212C cross\u2010coupling protocol. The Negishi C\u2212C cross\u2010coupling methodology allowed the formation of 2,7\u2010Fc2\u2010pyrene (7\u2009b) and 3,6\u2010Fc2\u20109,10\u2010dimethoxyphenanthrene (12).Compounds 1\u2010Fc\u2010 to 22.1(9)\u00b0). Parallel displaced \u03c0\u2010interactions were found for 10 and 12, and 10 and 12 exhibit a columnar stacking between the aromatic building blocks, leading to the formation of 1D (12) or 2D coordination polymers (10) along the a\u2010 (10) or b\u2010axis (12).The molecular structures of 3, 5\u2009a,b, 7\u2009a,b, 10, and 12, and 9\u2010ferrocenylphenanthrene for comparison, revealed a strong impact on the redox processes when bearing electron\u2010withdrawing or \u2010donating functionalities such as Br or OMe units. Within the series 3, 5\u2009a, and 7\u2009a, it could be shown that Br in position 1 influences the redox behavior most by shifting the potential cathodically from 50\u2005mV in 7\u2009a to \u2212130\u2005mV in 5\u2009a (vs. FcH/FcH+). The Fc group in position 1 in 3 (30\u2005mV vs. FcH/FcH+) is more easily to oxidize than the Fc in position 2 in 7\u2009a (E\u00b0\u2032=50\u2005mV). However, the 1,6\u2010 and 2,7\u2010substitution patterns of pyrenes 5\u2009b and 7\u2009b have no significant influence on the redox potentials and the redox splitting as was reported for Fc2\u2010naphthalenes,tert\u2010butyl\u20109\u2009H\u2010carbazol\u20109\u2010yl)\u2010, and 2,7\u2010((\u03b75\u2010C5Me5)(dppe)RuC\u2261C)2\u2010pyrenes.+ redox potential of 10 and 12 differ as expected for electron\u2010withdrawing (C(=O)) or \u2010donating (MeO) groups in positions 9 and 10 of the phenanthrene core. The redox splitting in the latter species is smaller than in pyrenes 5\u2009b and 7\u2009b, which is based on the closer proximity of the Fc ligands and the more electron\u2010poor \u03c0\u2010conjugated bridge in 10 and 12.Electrochemical studies of the Fc\u2010substituted aromatics 5\u2009b, 10, and 12, in situ UV/Vis/NIR studies were performed; 7\u2009b was excluded from these studies owing to its low solubility (see above). In the case of mono\u2010oxidized [10]+, the UV/Vis/NIR studies showed no IVCT absorption, which is due to the resonance structure of 10 and hence the redox separation is ascribed to electrostatic interactions. On the contrary, mixed\u2010valent [5\u2009b]+ and [12]+ showed IVCT absorptions, however, of low intensity, indicating weakly coupled systems according to the classification by Robin and Day, which is in good agreement with, for example, 2,7\u2010bispyrene.5\u2009b]+ appeared together with LMCT. Compound [12]+ showed a more intense and narrow IVCT absorption, indicating a stronger metal\u2013metal interaction than for 5\u2009b. This might originate from the higher electron density at the Fc groups, owing to the electron\u2010donating OMe functionalities in positions 9 and 10 at phenanthrene. Furthermore, the IVCT transition of [12]+ was investigated in two different solvents, exploring its solvatochromic characteristics. A \u0394\u03bdmax shift of 1000\u2005cm\u22121 was observed, confirming a class\u2005II classification according to Robin and Day.To confirm the metal\u2013metal interaction in 5\u2009a with 7\u2009a, it becomes apparent that positions 1 and 6 at pyrene are favorable over positions 2 and 7. Moreover, the higher electron density of phenanthrene 12 bearing electron\u2010donating functionalities resulted in a more intense LMCT than in Fc2\u2010pyrene 5\u2009b. These findings are in good agreement with, for example, ferrocenyl naphthalenes,In addition, within the spectroelectrochemical studies, LMCT transitions were observed that are shifted bathochromically depending on the characteristics of the substituents and the substitution pattern. Comparing the intensities of the LMCT transitions of 10 showed a complex solvation of the push\u2013pull system, reflecting the solvent properties such as hydrogen\u2010bond donor capacity, polarizability, and solvation, resulting in a bathochromic shift of \u03bbmax.Solvatochromic studies on 5\u2009b and SWCNTs was investigated by DFT calculations, which showed that the pyrene group of 5\u2009b is oriented parallel to the CNT surface based on optimized \u03c0\u2013\u03c0 interactions. The adsorption behavior of 5\u2009b is insignificantly influenced by the chirality or the diameter of the SWCNTs.The interaction between 5\u2009b to interact non\u2010covalently with different SWCNTs \u2010SWCNTs and semiconducting SWCNTs) was investigated by disentangling experiments. The debundeling with chirality\u2010enriched \u2010SWCNTs was performed for different sonication protocols (see the Supporting Information). For one parameter set, the dispersions containing SWCNTs and 5\u2009b in chloroform showed the typical S11 and the S22 transitions of disentangled SWCNTs in the UV/Vis/NIR spectra, which agree well with the reference spectra of a standard SWCNT dispersion, indicating a direct non\u2010covalent interaction between 5\u2009b and the SWCNT sidewalls.Based on the DFT calculations, the ability of 5\u2009b with semiconducting SWCNTS was studied (Gen2). As a result thereof, it was found that 5\u2009b is able to disentangle SWCNTs as evident from the appearance of the S22 and S33 transitions in the UV/Vis/NIR spectra.With regard to the possible use of functionalized SWCNTs, integrated in CNT\u2010FETs, the interaction of 5\u2009b in an exohedral fashion at the SWCNT sidewalls was proven by CV by using graphene paper as the working electrode, which showed a Fc/Fc+ redox event at 380\u2005mV. This was further strengthened by the electrochemical investigations of solely 5\u2009b, showing no ferrocenyl\u2010based redox processes. At a potential of 800\u2005mV, the irreversible oxidation of the pyrene core occurred, resulting in polymerization and subsequent formation of polypyrene oligomers.The attachment of The debundeling and functionalization of SWCNTs by redox\u2010active Fc\u2010pyrenes in a non\u2010covalent manner is conceivable when using the 1,6\u2010substitution pattern at pyrene.5\u2009b could be used in nanoelectronic application scenarios.Based on these findings, compound\u2005All synthesis procedures were performed under an atmosphere of argon with the solvents degassed prior to use. All reagents were obtained from commercial suppliers and used without further purification. The SWCNTs solid material was purchased from a commercial supplier \u2010SWCNTs, batch number #MKB76336V) and used without further treatment. A commercial SWCNT dispersion, NanoIntegris IsoSol S\u2010100\u00ae, batch SP31\u2010176, was purchased from commercial suppliers and used without any further purification. Ferrocene boronic acid was synthesized from lithioferrocene1H and 125.7\u2005MHz for 13C{1H} in the Fourier transform mode at ambient temperature. Chemical shifts are reported in \u03b4 (ppm) downfield from tetramethylsilane with the solvent as the reference signal . FTIR spectra were recorded by using a Thermo Nicolet IR 200 instrument. The melting points were determined with a Gallenkamp MFB 595 010\u2009m melting point apparatus. Elemental analyses were performed by applying a Thermos FlashAE 1112 instrument. High\u2010resolution mass spectra were recorded with a Bruker Daltonite micrOTOF\u2010QII spectrometer . UV/Vis spectra between 210 and 1010\u2005nm were recorded with a Carl Zeiss MCS 400 spectrometer utilizing CLD 300 (210\u2013600\u2005nm) and CLX 11 lamps (300\u20131010\u2005nm). UV/Vis/NIR spectra of the SWCNT dispersions between 350\u2005nm and 1300\u2005nm were recorded by using a Shimadzu UV\u20103100 PC absorption spectrometer on liquids in quartz cuvettes . For sonication, an Elmasonic P 30 H ultrasonic bath and a Bandelin KE76 tip sonotrode were used. Centrifugation of the dispersions was performed at a Sigma 3\u201330\u2005K lab centrifuge in Oak Ridge tubes with 10\u2005mL nominal volume.NMR spectra were recorded with a Bruker Avance III 500 spectrometer operating at 500.3\u2005MHz for 3, 5\u2009a,b, 7\u2009a,b, 10, and 12 (1.0\u2005mmol\u2009L\u22121 [NnBu4][B(C6F5)4] as the supporting electrolyte) in dichloromethane were performed in a dried, argon\u2010purged cell at 25\u2009\u00b0C. For the measurements, a three\u2010electrode cell containing a Pt auxiliary electrode, a glassy\u2010carbon working electrode (3.0\u2005mm diameter), and an Ag/Ag+ (0.01\u2005mmol\u2009L\u22121 [AgNO3]) reference electrode fixed on a Luggin capillary was used. The working electrode was pretreated by polishing it with a MicroFloc first with 1\u2005\u03bcm and then with a 0.25\u2005\u03bcm diamond paste. The reference electrode was constructed from a silver wire inserted in a 0.01\u2005mmol\u2009L\u22121 [AgNO3] and a 0.1\u2005mol\u2009L\u22121 [NnBu4][B(C6F5)4] acetonitrile solution in a Luggin capillary with a Vycor tip. This Luggin capillary was inserted into a second Luggin capillary containing a 0.1\u2005mol\u2009L\u22121 [NnBu4][B(C6F5)4] dichloromethane solution and a Vycor tip. Experiments under the same conditions showed that all reduction and oxidation potentials were reproducible within \u00b15\u2005mV. Experimental potentials were referenced against an Ag/Ag+ reference electrode, but the presented results are referenced against ferrocene as an internal standard as required by IUPAC.\u22121 decamethylferrocene (Fc*). Data were processed with a Microsoft Excel worksheet to set the formal reduction potentials of the FcH/FcH+ couple to 0.0\u2005V. Under our conditions, the Fc*/Fc*+ couple appeared at \u2212619\u2005mV vs. FcH/FcH+, \u0394Ep=60\u2005mV, whereas the FcH/FcH+ couple itself was at 220\u2005mV vs. Ag/Ag+, \u0394Ep=61\u2005mV.5\u2009b@SWCNT (Gen2) nanoconjugates, a three\u2010electrode arrangement containing a Pt auxiliary electrode and an Ag/AgCl reference electrode (consisting of a silver wire covered with a thin film of AgCl) fixed in a glass tube (providing a pipetting controller for the supply of the electrolyte) at 25\u2009\u00b0C was used.3[Fe(CN)6] (5\u2005mm aqueous solution) in 1\u2009m aqueous KCl solution (0.358\u2005V).Electrochemical measurements of \u22121 solutions of 3, 5, 7\u2009a, 10, 12, or 9\u2010ferrocenylphenanthrene in anhydrous dichloromethane containing 0.1\u2005mol\u2009L\u22121 of [NnBu4][B(C6F5)4] as the supporting electrolyte were performed in an OTTLE \u03f5max \u00b1100\u2005L\u2009mol\u22121\u2009cm\u22121, \u03bdmax \u00b150\u2005cm\u22121, and \u0394\u03bd1/2 \u00b150\u2005cm\u22121.Spectroelectrochemical UV/Vis/NIR measurements of 2.0\u2005mmol\u2009LK\u03b1 radiation or CuK\u03b1 radiation at 110\u2005K by using oil\u2010coated shock\u2010cooled crystals. The structures were solved by direct methods and refined by full\u2010matrix least\u2010squares procedures on F2.Diffraction data were collected with an Oxford Gemini S diffractometer using graphite\u2010monochromated MoEads) of 5\u2009b on CNT is calculated by Eads=E5\u2009b/CNT\u2212E5\u2009b\u2212ECNT, where E5\u2009b/CNT, E5\u2009b, and ECNT denote the total energies of the optimized structures of 5\u2009b adsorbed on CNT, gaseous 5\u2009b, and isolated CNT, respectively.Density functional theory (DFT) calculations were performed with periodic models by using the CASTEP code.2] (1\u2005mol\u2009%), ferrocene boronic acid , K3PO4\u22c5H2O , and the respective aryl halide . Anhydrous toluene (15\u2005mL) was added. The reaction mixture was stirred for 5\u2005min at ambient temperature and then heated to reflux for 24\u2005h. After cooling to ambient temperature, the reaction mixture was filtered through a pad of silica. Afterwards, all volatiles were removed by evaporation. Purification was realized by column chromatography by using different dichloromethane/hexane mixtures (see below).A three\u2010necked 100\u2005mL flask was charged with +; elemental analysis calcd for C26H17BrFe (465.16\u2005g\u2009mol\u22121): C 67.13, H 3.68, found: C 67.69, H 4.76. Crystal data for 5\u2009a: C26H17BrFe, Mr=465.15\u2005g\u2009mol\u22121, monoclinic, I2/a, \u03bb=0.71073\u2005\u00c5, a=18.4055(14)\u2005\u00c5, b=7.4788(6)\u2005\u00c5, c=26.788(2)\u2005\u00c5, \u03b2=96.988(7)\u00b0, V=3659.9(5)\u2005\u00c53, Z=8, \u03c1calcd=1.688\u2005mg\u2009cm\u22123, \u03bc=3.016\u2005mm\u22121, T=116.8(6)\u2005K, \u03b8 range 3.565\u201324.992\u00b0, 8764 reflections collected, 3199 independent reflections (Rint=0.0352), R1=0.0327, wR2=0.0701 (I>2\u03c3(I)).Compound\u20055\u2009b: Yield: 72.3\u2005mg . M.p.: 176\u2009\u00b0C (decomposition); 1H\u2005NMR (CDCl3): \u03b4=4.22 , 4.49 , 4.84 , 8.02 , 8.10 , 8.39 , 8.73\u2005ppm ; 13C\u2005NMR (CDCl3): \u03b4=68.7 (C5H4), 69.9 (C5H5), 71.1 (C5H4), 87.6 , 124.1 (C16H8), 125.1 (C16H8), 125.4 (C16H8), 127.0 (C16H8), 129.0 (C16H8), 129.3 (C16H8), 129.7 (C16H8), 134.0\u2005ppm (C16H8); IR (KBr): \u03bd=2964 (m), 2925 (w), 2855 (w), 1601 (w), 1496 (w), 1456 (w), 1428 (w), 1409 (w), 1373 (w), 1262 (s), 1211 (w), 1105 (s), 1074 (s), 1022 (s), 881 (m), 866 (m), 854 (m), 804 (s), 684 (m), 668\u2005cm\u22121 (m); HR\u2010MS (ESI\u2010TOF) m/z calcd for C36H26Fe2: 570.0729; found: 570.0734 [M]+; elemental analysis calcd for C36H26Fe2 (570.28\u2005g\u2009mol\u22121): C 75.82, H 4.60; found: C 75.56, H 4.52. Crystal data for 5\u2009b: C36H26Fe2, Mr=570.27\u2005g\u2009mol\u22121, monoclinic, P21/n, \u03bb=0.71073\u2005\u00c5, a=14.3748(18)\u2005\u00c5, b=8.6113(9)\u2005\u00c5, c=20.585(2)\u2005\u00c5, \u03b2=105.022(12)\u00b0, V=2461.1(5)\u2005\u00c53, Z=4, \u03c1calcd=1.539\u2005mg\u2009cm\u22123, \u03bc=1.204\u2005mm\u22121, T=116.95(10)\u2005K, \u03b8 range 3.043\u201324.996\u00b0, 5798 reflections collected, 5798 independent reflections (Rint=0.0655), R1=0.0771, wR2=0.1997 (I>2\u03c3(I)).7\u2009a was prepared according to the general synthetic methodology described above by using 2,7\u2010dibromopyrene . Compound\u20057\u2009a was separated by column chromatography by using hexane/dichloromethane mixtures (v/v) starting from 9:1 to 4:1 (7\u2009a). After evaporation of all volatiles, the received solid was recrystallized from acetone at \u221220\u2009\u00b0C. Compound\u20057\u2009a was isolated as an orange solid. Yield: 37\u2005mg . M.p.: 223\u2009\u00b0C; 1H\u2005NMR (CDCl3): \u03b4=4.08 , 4.46 , 4.96 , 7.96 , 8.07 , 8.26\u2005ppm ; 13C\u2005NMR (CDCl3): \u03b4=67.2 (C5H4), 69.7 (C5H4), 70.0 (C5H5), 85.3 , 119.7 (C16H8), 123.1 (C16H8), 123.4 (C16H8), 123.5 (C16H8), 126.7 (C16H8), 127.3 (C16H8), 128.7 (C16H8), 131.1 (C16H8), 132.6 (C16H8), 138.0\u2005ppm (C16H8); IR (KBr): \u03bd=3036 (w), 2924 (w), 2852 (w), 1597 (s), 1554 (m), 1487 (m), 1429 (m), 1409 (m), 1299 (m), 1245 (s), 1217 (w), 1152 (m), 1145 (m), 1105 (s), 1037 (m), 1030 (m), 998 (s), 929 (m), 898 (m), 878 (s), 867 (s), 858 (s), 853 (s), 846 (s), 830 (s), 820 (s), 803 (s), 762 (s), 707 (s), 669\u2005cm\u22121 (s); HR\u2010MS (ESI\u2010TOF) m/z calcd for C26H17BrFe: 463.9859; found: 463.9849 [M]+; elemental analysis calcd for C26H17BrFe (465.16\u2005g\u2009mol\u22121): C 67.13, H 3.68; found: C 67.25, H 3.71. Crystal data for 7\u2009a: C26H17BrFe, Mr=465.15\u2005g\u2009mol\u22121, monoclinic, P21/c, \u03bb=0.71073\u2005\u00c5, a=10.3922(8)\u2005\u00c5, b=13.9915(11)\u2005\u00c5, c=12.8138(11)\u2005\u00c5, \u03b2=99.217(3)\u00b0, V=1839.1(3)\u2005\u00c53, Z=4, \u03c1calcd=1.680\u2005mg\u2009cm\u22123, \u03bc=3.001\u2005mm\u22121, T=100\u2005K, \u03b8 range 3.525\u201324.997\u00b0, 26\u2009828 reflections collected, 3234 independent reflections (Rint=0.0982), R1=0.0268, wR2=0.0707 (I>2\u03c3(I)).Compound\u200510 was synthesized in accordance with the general synthesis procedure described above by using 3,6\u2010dibromophenathren\u20109,10\u2010dione . Compound\u200510 was separated by column chromatography by using first hexane and then dichloromethane/ethylacetate mixtures (v/v) of ratios 4:1 and 3:1 (10). After evaporation of all volatiles, compound\u200510 was obtained as a green solid. Yield: 136\u2005mg . M.p.: 123\u2009\u00b0C; 1H\u2005NMR (CDCl3): \u03b4=4.13 , 4.55 , 4.86 , 7.56 , 8.05 , 8.15\u2005ppm ; 13C\u2005NMR (CDCl3): \u03b4=67.6 (C5H4), 70.3 (C5H5), 70.9 (C5H4), 83.0 , 120.6 (C14H6), 127.1 (C14H6), 129.0 (C14H6), 130.9 (C14H6), 136.0 (C14H6), 149.5 (C14H6), 180.1\u2005ppm (CO); IR (KBr): \u03bd=2959 (m), 2925 (s), 2854 (m), 1774 (w), 1658 (m), 1590 (s), 1465 (m), 1416 (m), 1262 (s), 1105 (s), 1029 (s), 924 (m), 807 (s), 731\u2005cm\u22121 (m); elemental analysis calcd for C34H24Fe2O2 (576.25\u2005g\u2009mol\u22121): C 70.87, H 4.20; found: C 70.76, H 4.11. Crystal data for 10: C35H26Cl2Fe2O2, Mr=661.16\u2005g\u2009mol\u22121, monoclinic, P21/c, \u03bb=0.71073\u2005\u00c5, a=10.1023(5)\u2005\u00c5, b=38.6119(18)\u2005\u00c5, c=13.9755(8)\u2005\u00c5, \u03b2=93.989(5)\u00b0, V=5438.2(5)\u2005\u00c53, Z=8, \u03c1calcd=1.615\u2005mg\u2009cm\u22123, \u03bc=1.297\u2005mm\u22121, T=129.9(4)\u2005K, \u03b8 range 3.023\u201325.999\u00b0, 34\u2009953 reflections collected, 10\u2009644 independent reflections (Rint=0.0537), R1=0.0819, wR2=0.1943 (I>2\u03c3(I)).Compound\u2005m solution of tert\u2010butyllithium in pentane was added dropwise at \u221230\u2009\u00b0C to ferrocene and KOtBu dissolved in tetrahydrofuran (20\u2005mL). After 1\u2005h of stirring at this temperature, anhydrous [ZnCl2 2\u2009thf] was added in a single portion. The solution was kept for 1\u2005h at \u221230\u2009\u00b0C and an additional hour at 25\u2009\u00b0C. Afterwards, [PdCl2(dppf)] and the respective dibromoarenes were added in a single portion and the reaction solution was stirred for 24\u2005h at 60\u2009\u00b0C. After evaporation of all volatiles, the precipitate was dissolved in dichloromethane (200\u2005mL) and washed thrice with 100\u2005mL portions of water. The organic phase was dried over MgSO4 and the solvent was removed under oil\u2010pump vacuum. The remaining solid was purified by column chromatography by using different dichloromethane/hexane mixtures. All volatiles were removed under reduced pressure. The title compounds were obtained as solids.A 1.9\u20097\u2009b was synthesized according to the general synthetic methodology for the Negishi C\u2212C cross\u2010coupling protocol by using 2,7\u2010dibromopyrene . Compound\u20057\u2009b was separated by column chromatography by using hexane/dichloromethane eluent mixtures (v/v) starting from 9:1 to 1:9 (7\u2009b). The obtained solid was recrystallized from acetone at \u221220\u2009\u00b0C. Compound\u20057\u2009b was isolated as a red solid. Yield: 15\u2005mg . M.p.: 265\u2009\u00b0C (decomposition); 1H\u2005NMR (CDCl3): \u03b4=4.08 , 4.45 , 4.96 , 8.04 , 8.24\u2005ppm ; IR (KBr): \u03bd=3116 (w), 3092 (w), 1606 (m), 1424 (m), 1407 (m), 1382 (m), 1103 (s), 1030 (s), 1000 (s), 932 (m), 881 (m), 829 (m), 806 (m), 714 (m), 668\u2005cm\u22121 (w); elemental analysis calcd for C36H26Fe2 (570.28\u2005g\u2009mol\u22121): C 75.82, H 4.60; found: C 76.63, H 4.43. Crystal data for 7\u2009b: C36H26Fe2, Mr=570.27\u2005g\u2009mol\u22121, monoclinic, P21/c, \u03bb=1.54184\u2005\u00c5, a=11.0921(3)\u2005\u00c5, b=7.8868(2)\u2005\u00c5, c=14.2973(4)\u2005\u00c5, \u03b2=100.294(2)\u00b0, V=1230.61(6)\u2005\u00c53, Z=2, \u03c1calcd=1.539\u2005mg\u2009cm\u22123, \u03bc=9.630\u2005mm\u22121, T=100\u2005K, \u03b8 range 4.051\u201365.912\u00b0, 11\u2009449 reflections collected, 2128 independent reflections (Rint=0.0443), R1=0.0375, wR2=0.0961 (I>2\u03c3(I)).Compound\u200511; 396\u2005mg, 0.5\u2005mmol). Compound\u200512 was separated by column chromatography by using hexane/dichloromethane eluent mixtures (v/v) starting from 9:1 to 1:9 (12). After evaporation of all volatiles, 12 was obtained as an orange solid. Yield: 100\u2005mg . M.p.: 169\u2009\u00b0C; 1H\u2005NMR (CDCl3): \u03b4=4.10 , 4.12 , 4.43 , 4.86 , 7.80 , 8.16 , 8.68\u2005ppm ; 13C\u2005NMR (CDCl3): \u03b4=61.2 (OCH3), 67.0 (C5H4), 69.4 (C5H5), 69.9 (C5H4), 86.0 , 119.4 (C14H6), 122.3 (C14H6), 126.1 (C14H6), 127.8 (C14H6), 128.5 (C14H6), 136.9\u2005ppm (C14H6); IR (KBr): \u03bd=2925 (s), 2854 (s), 1744 (m), 1700 (m), 1684 (m), 1653 (m), 1607 (s), 1559 (m), 1540 (m), 1507 (m), 1458 (s), 1446 (s), 1419 (m), 1337 (m), 1311 (s), 1189 (m), 1105 (s), 1092 (s), 1061 (s), 1027 (m), 983 (s), 885 (m), 874 (s), 707 (w), 668 (w), 661\u2005cm\u22121 (w); elemental analysis calcd for C36H30Fe2O2 (606.32\u2005g\u2009mol\u22121): C 71.31, H 4.99; found: C 70.50, H 4.89. Crystal data for 12: C36H30Fe2O2, Mr=606.30\u2005g\u2009mol\u22121, orthorhombic, Pnn2, \u03bb=1.54178\u2005\u00c5, a=19.680(2)\u2005\u00c5, b=6.3428(8)\u2005\u00c5, c=10.6327(17)\u2005\u00c5, V=1327.3(3)\u2005\u00c53, Z=2, \u03c1calcd=1.517\u2005mg\u2009cm\u22123, \u03bc=9.021\u2005mm\u22121, T=100(2)\u2005K, \u03b8 range 4.493\u201364.897\u00b0, 23\u2009836 reflections collected, 2100 independent reflections (Rint=0.1099), R1=0.1412, wR2=0.4059 (I>2\u03c3(I)).The title compound was prepared according to the general synthesis protocol for the Negishi C\u2212C cross\u2010coupling by using 3,6\u2010dibromo\u20109,10\u2010dimethoxyphenanthrene (Master5\u2009b was prepared by dissolving 5\u2009b (1.86\u2005mg) in chloroform . Afterwards, the obtained solution was treated by bath sonication for 10\u2005min.The master solution Master5\u2009b (800\u2005\u03bcL) in chloroform .The mixture FC00 was obtained by diluting The mixture FC01 was prepared by dispersing SWCNTs solid material (0.82\u2005mg) in FC00 (6\u2005mL) and applying bath sonication for 30\u2005min, followed by centrifugation .The mixture FC02 was prepared by dispersing SWCNTs solid material (0.75\u2005mg) in FC00 (6\u2005mL) and applying tip sonication for 30\u2005min, followed by centrifugation .\u22121.Dispersions FC01 and FC02 were prepared with a SWCNT solid content of (0.128\u00b10.009)\u2005mg\u2009mL3) was obtained by pouring SWCNTs into CHCl3 and employing bath sonication for 30\u2005min followed by centrifugation . The SWCNT solid content was kept to (0.128\u00b10.009)\u2005mg\u2009mL\u22121.The reference dispersion SWCNTs (CHCl2O) was obtained by adapting previously reported sonication procedures.\u22121.The reference dispersion of SWCNTs (HH\u2010fluoren\u20102\u2010yl)aryl\u2010based polymer (PFH\u2010R)5\u2009b dissolved in toluene to attach 5\u2009b at those parts of the SWCNT sidewalls depleted from PFH\u2010R. To apply UV/Vis/NIR analysis for this step, the bucky paper obtained this way was manually delaminated from the filter by using standard lab cutlery. The slices of the SWCNT solid were then collected in a new glass vial (10\u2005mL screw\u2010cap) and re\u2010dispersed in toluene (2\u2005mL) by using a bath sonicator . The obtained liquid obtained by this procedure (Gen2) was subjected to the UV/Vis/NIR analysis and compared with basic spectra of 5\u2009b in toluene, just diluted NanoIntegris IsoSol S\u2010100\u00ae (Gen0) and a reference sample obtained directly by dissolving a bucky paper derived from NanoIntegris IsoSol S\u2010100\u00ae without the washing (toluene) and flushing (5\u2009b in toluene) steps (Gen1).The commercial SWCNT dispersion, stabilized by a 9\u2010 for all new compounds are given. This material is available free of charge via the Internet. CCDC\u2005https://www.ccdc.cam.ac.uk/services/strctures?id=doi:10.1002/chem.201904450 contain the supplementary crystallographic data for this paper. These data are provided free of charge by http://www.ccdc.cam.ac.uk/.Additional structural data, cyclic voltammograms, square wave voltammograms, and UV/Vis/NIR spectra as well as spectroscopic details should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "In contrast, 1,7\u2010bis(1\u2010phospholano)heptane (1b) formed coordination polymers 4a (Cl) and 4b (Br) with bridging bis\u2010phospholane and halido ligands. A unique paddle wheel\u2010type metallacryptand structure 5 was obtained from 1a and silver(I) bromide in a 2:3 reaction (M:L). All complexes were fully characterized by NMR, IR spectroscopy, mass spectrometry, and X\u2010ray crystallography.In a 2:2 reaction with silver(I) chloride or bromide, 1,5\u2010bis(1\u2010phospholano)pentane ( John Wiley & Sons, Ltd. Gr\u00fcttner and Krause. They synthesized 1\u2010phenylphospholane by reacting the di\u2010Grignard reagent of 1,4\u2010dibromobutane with dichlorophenylphosphine.Brunner et al. published the synthesis of the chiral 3,4\u2010disubstituted correspondent.Wilson et al.Burk et al.Baccolini et al.H\u2010phospholane and P\u2010substituted phospholanes can be obtained using this method. In 2009, the same group reported the successful synthesis of chiral alkylphospholanes.Phospholanes are five\u2010membered cyclic phosphines with good \u03c3\u2010donor properties, which make them especially interesting as ligands in transition metal catalysis. Their ring structure and well\u2010defined geometry are key features, which set them apart from other phosphines.Burk and co\u2010workers.Burk published a paper entitled \u201cModular Phospholane Ligands in Asymmetric Catalysis\u201d, which shows the ten\u2010year development of ligands based on the trans\u20102,5\u2010disubstituted phospholane moiety.Clark and Landis comprehensively summarized the following developments in this field.The real success story of the phospholane ligands started, however, in 1990 with the synthesis of bis\u2010phospholanoethane (BPE) and DuPhos by cis complexes with catalytically active metals.Emrich and Jolly,Oisaki et al.4 as a metal complex precursor.3CN)4]BF4.These chiral bis\u2010phospholanes are usually based on rigid spacers that allow them to act as chelate ligands forming 1a)1b)We have performed an in\u2010depth study of the coordination behavior of these flexible bis\u2010phospholane ligands in silver(I) halide complexes and report here the synthesis, characterization, and structural elucidation of complexes based on 1,5\u2010bis(1\u2010phospholano)pentane (1a)31P{1H} NMR spectrum of the crude reaction mixture showed the expected downfield shift from the free ligands (ca. \u201327 ppm)2 and 3 suitable for X\u2010ray structure analysis were obtained at room temperature by slow diffusion of n\u2010hexane into a DCM/toluene solution. The results showed two different coordination modes: A metallacycle 2 with intramolecular chlorido bridges (Table 2 [245.5(2) and 245.7(2) pm] which are more than 8 pm longer than those in the corresponding AgBF4 complex.2Cl2 moiety forms an almost perfect square as shown by Ag\u2013Cl bond lengths of 271.1(2) and 270.9(2) pm and a Cl1\u2013Ag1\u2013Cl1' bond angle of 90.00(5)\u00b0. A similar structure was reported for the 2:2 complexes of 1,5\u2010bis(diphenylphosphanyl)pentane (dpppn) with AgX by Effendy et al.Cassel.2, which is to be expected given the electron\u2010withdrawing effect of the phenyl groups of the triphenylphosphine.Complex 31P{1H} NMR spectrum of complex 2 in CDCl3 shows a singlet at \u03b4 = \u20135.7 ppm. Coupling with the NMR\u2010active silver isotopes 107Ag (NA = 51.84\u2009%) or 109Ag is not observed. The lack of coupling in solution at room temperature was also observed by Streitberger et al.Effendy and co\u2010workers who carried out more thorough NMR studies on silver(I) bis(diphenyl)phosphine complexes.2 in solution is evidenced by the ESI(+) mass spectrum which showed the [M\u2013Cl]+ molecular ion at m/z = 739.1.The 3 crystallizes in the monoclinic space group C2/c with four molecules in the unit cell. The solid\u2010state structure shows two molecules of 1,5\u2010bis(1\u2010phospholano)pentane bridging two silver(I) cations forming a frame\u2010like macrocycle and 237.02(8) pm but a significantly larger P\u2013Ag\u2013P bond angle of 176.89(1)\u00b0 for a distinct frame\u2010like structure obtained using AgBF4 cations of neighboring metallamacrocycles interact with an additional AgBr moiety, thus forming Ag3Br3 assemblies phosphine complexes. Often they form tetrameric cubane\u2010like structures (Ag4Br4), when simple monodentate phosphine ligands are employed.Effendy et al. reported a tetranuclear 2:1 complex , which is formed when AgX and bis(diphenylphosphanyl)methane (dppm) are reacted in a 2:1 ratio. The structure of the complex includes an Ag4Br4 assembly with a \u201cstep\u201d conformation.Schubert et al. found that dppm and AgBr can also form a 3:3 complex bearing an [Ag3Br2]+ cluster, while bis(diphenylphosphanyl)methylamine (dppa) forms a dimeric 2:4 complex in which an [Ag4Br2]2+ octahedron is present.3Br3 assembly to the one found in complex 3 was assumed to bridge adjacent macrocycles in a silver(I) complex based on 2,5\u2010bis(diphenylphosphinomethyl)thiophene (dpmt). However, only the crystal structure of the iodine homologue was reported.3 shows that Ag3Br3 bicycles are bridged by a set of two 1,5\u2010bis(1\u2010phospholano)pentane ligands, building a polymeric chain along (101) .Ag31P{1H} NMR spectrum of complex 3 in CDCl3 shows a singlet at \u03b4 = \u20138.5 ppm. The ESI(+) mass spectrum showed peaks at m/z = 1590.6, 1402.8, 1158.7, and 1215.0, corresponding to the fragments [M+L+2Ag+Br]+, [M+L+Ag]+, [M+Ag]+, and [3 L+3Ag+2Br]+, respectively. The peaks at m/z = 970.9 (base peak) and 780.0 corresponding to fragments [M\u2013Br]+ and [M\u2013AgBr2]+, respectively, are proof that the frame\u2010like metallamacrocycle is stable in solution .The 1b)1a. The reaction mixtures were shielded from light. After stirring at room temperature for four hours, the 31P{1H} NMR spectra of the crude reaction mixture showed the expected downfield shift with signals around \u20137 ppm. Crystals of the final products 4a and 4b are marginally shorter than in complex 2. One of the Ag\u2013Cl bonds (Ag1\u2013Cl2) is ca. 6 pm shorter than the one in 2, causing the Ag2Cl2 ring to slightly deviate from the ideal square arrangement, with angles ranging from 85.54(4) to 93.24(3)\u00b0 (Table Ag\u2013P bond lengths in 4a forms a (001) layer structure in which the silver(I) cations are linked in series by chlorine atoms, while the bis\u2010phospholane ligands are arranged in a zigzag fashion. The layers are formed by two distinct interwoven polymeric chains mass spectrum showed the base peak at m/z = 795 corresponding to the fragment [L2Ag2Cl]+ (L = ligand 1b).The 4b crystallizes in the monoclinic space group P21/c. The solid\u2010state structure layer to 138.70(3)\u00b0, Table 3 alkane ligands when comparing open macrocycles [P\u2013Ag\u2013P 144.30(7)\u00b0] to species in which halido bridging was observed [P\u2013Ag\u2013P 131.24(9)\u00b0].4b + (L = ligand 1b). Two other peaks corresponding to species with higher m/z values were observed but could not be attributed and probably correspond to oligomeric fragments.The 1a,1b and two equivalents of AgBr were performed under similar conditions as described for the 1:1 reactions. The 31P{1H} NMR spectrum of the crude reaction mixtures showed, in each case, one downfield\u2010shifted broad singlet. Suitable crystals for X\u2010ray structure analysis were obtained at room temperature by layering a DCM/toluene (for ligand 1a) or DCM (for ligand 1b) solution of the crude product with n\u2010hexane. X\u2010ray structure analysis showed that in the case of ligand 1b the same coordination polymer 4b was formed as in the 1:1 reaction, while ligand 1a yielded the metallacryptand 5 cations are bridged by three 1,5\u2010bis(1\u2010phospholano)pentane ligands in a trigonal\u2010planar fashion forming a triangular prism (paddle wheel) with Ag\u2013P distances ranging from 241.88(9) to 251.48(9) pm and P\u2013Ag\u2013P angles ranging from 104.74(3) to 137.51(3)\u00b0 and 248.5(1) pm, respectively]. Ag\u2013Br bond lengths [278.90(4) and 281.84(5) pm] are comparable to those previously observed in 4b.Complex 31P{1H} NMR studies reported by Dean et al. (1987) and later by Peringer et al. (1988) postulated the formation of complexes [Ag2(\u03bc\u2010dppm)3](AsF6)22(\u03bc\u2010dppm)3](CF3SO3)2,Hong et al. published the structure of [Ag2(\u03bc\u2010dppm)3](NO3)2 and revealed its paddle wheel topology.5 is not only the first such example bearing bis\u2010phospholane ligands but also the first silver(I) bis\u2010phosphine paddle wheel\u2010type metallacryptand.Silver(I) complexes with an M:L ratio of 2:3 and with bridging bis(phosphanyl)alkane ligands are rare. 1a) and 1,7\u2010bis(1\u2010phospholano)heptane (1b), gave macrocycles, coordination polymers and paddle wheel\u2010type metallacryptands depending on the stoichiometry, length of alkylene spacer (C5 or C7) and counter anion. These complexes are further proof that these simple and highly flexible bis\u2010phospholane ligands have extremely versatile coordination properties.The reaction of silver(I) chloride or bromide with two highly flexible bis\u2010phospholane ligands, 1,5\u2010bis(1\u2010phospholano)pentane (1b)\u03b4) of 1H, 13C, and 31P are reported in parts per million (ppm) at 400.12, 100.63, and 162.02 MHz, respectively, with tetramethylsilane as internal standard and referencing to the unified \u039e scale.J are given in Hertz. The numbering scheme for assignment of 1H NMR spectra is given in Scheme \u20131. Wavenumbers \u03bd\u0303 are reported in cm\u20131. Mass spectra were recorded with an ESQUIRE 3000 plus spectrometer. Elemental analyses were carried out with a Heraeus VARIO EL oven. Melting points were measured in sealed capillaries by using a variable heater from Gallenkamp.All reactions and crystallization of compounds were carried out in a nitrogen atmosphere by using standard Schlenk techniquesSynthesis of Bisdisilver(I) dichloride (2): AgCl was added as a solid to a stirred solution of 1a in 10 mL DCM. The Schlenk flask was immediately wrapped in aluminum foil and the reaction mixture was stirred at room temperature for 4 h, filtered and all volatiles were removed in vacuo. Afterwards, the white residue was washed with n\u2010hexane (3\u2009\u00d7\u20092 mL) to yield 2 as a white solid. Crystals suitable for single\u2010crystal X\u2010ray diffraction were obtained from a DCM/toluene mixture layered with n\u2010hexane at room temperature. M.p. 129 \u00b0C. 1H NMR (CDCl3): \u03b4 = 2.19\u20132.03 , 1.92\u20131.72 , 1.72\u20131.59 , 1.50\u20131.48 ppm . 13C{1H} NMR (CDCl3): \u03b4 = 32.3\u201332.0 , 28.0\u201327.9 , 27.2 (s), 27.1\u201326.9 , 26.3\u201325.7 ppm . 31P{1H} NMR (CDCl3): \u03b4 = \u20135.7 ppm (s). IR: \u03bd\u0303 = 2937 , 2915 , 2855 , 1455 , 1436 , 1113 (w), 843 (w), 689 cm\u20131 (w). MS (ESI(+), DCM/acetonitrile): m/z = 739.1 [M\u2013Cl]+, 597.2 [AgL2]+, 351.1 [AgL]+; L = 1a. C26H52Ag2Cl2P4: calcd. C 40.28, H 6.76\u2009%; found C 40.29, H 6.82\u2009%.Synthesis of Poly{bistrisilver(I) tribromide} (3): AgBr was added as a solid to a stirred solution of 1a in 10 mL DCM. The Schlenk flask was immediately wrapped in aluminum foil and the reaction mixture was stirred at room temperature for 4 h, filtered and all volatiles were removed in vacuo. Afterwards the white residue was washed with n\u2010hexane (3\u2009\u00d7\u20092 mL) to remove excess ligand, yielding 3 as a white solid. Crystals suitable for single\u2010crystal X\u2010ray diffraction were obtained from a DCM solution layered with n\u2010hexane at room temperature. M.p. 195 \u00b0C. 1H NMR (CDCl3): \u03b4 = 2.20\u20132.04 , 1.91\u20131.76 , 1.75\u20131.55 ppm . 13C{1H} NMR (CDCl3): \u03b4 = 27.6\u201327.3 , 27.1 (s), 26.3\u201326.0 , 26.0\u201325.8 ppm . 31P{1H} NMR (CDCl3): \u03b4 = \u20138.5 ppm (s). IR: \u03bd\u0303 = 2934 , 2919 , 2845 , 1444 , 1407 , 1112 (w), 1023 (w), 845 (w), 708 cm\u20131 (w). MS (ESI(+), DCM/MeOH): m/z = 1590.6 [M+L+2Ag+Br]+, 1402.8 [M+L+Ag]+, 1158.7 [M+Ag]+, 1215.0 [Ag3L3Br2]+, 970.9 [M\u2013Br]+, 780.0 [M\u2013AgBr2]+; M = [Ag3L2Br3], L = 1a. C26H52Ag3Br3P4: calcd. C 29.69, H 4.98\u2009%; found C 29.66, H 5.01\u2009%.Synthesis of Poly{bissilver(I) chloride} (4a): AgCl was added as a solid to a stirred solution of 1b in 10 mL DCM. The Schlenk flask was immediately wrapped in aluminum foil and the reaction mixture stirred at room temperature for 4 h, filtered and all volatiles were removed in vacuo. Afterwards the white residue was washed with n\u2010hexane (3\u2009\u00d7\u20092 mL) to yield 4a as a white solid. Crystals suitable for single\u2010crystal X\u2010ray diffraction were obtained from a DCM/THF mixture layered with n\u2010hexane at room temperature. M.p. 103 \u00b0C. 1H NMR (CDCl3): \u03b4 = 2.12\u20131.99 , 1.85\u20131.66 , 1.64\u20131.53 , 1.51\u20131.14 . 13C{1H} NMR (CDCl3): \u03b4 = 31.1\u201330.5 , 29.2 (s), 28.3\u201327.8 , 27.2 (s), 27.1\u201326.8 , 26.3\u201325.8 ppm . 31P{1H} NMR (CDCl3): \u03b4 = \u20135.3 ppm (s). IR: \u03bd\u0303 = 2962 , 2927 , 2856 , 1446 , 1261 (s), 1105 (s), 1024 (s), 802 (s), 720 cm\u20131 (w). MS (ESI(+), DCM/MeOH): m/z = 795.0 [Ag2L2Cl]+, 651.0 [AgL2]+, 379.0 [AgL]+; L = 1b. C15H30AgClP2: calcd. C 43.34, H 7.27\u2009%; found C 43.30, H 7.21\u2009%.Synthesis of Poly{bissilver(I) bromide} (4b): AgBr was added as a solid to a stirred solution of 1b in 10 mL DCM. The Schlenk flask was immediately wrapped in aluminum foil and the reaction mixture was stirred at room temperature for 4 h, filtered and all volatiles were removed in vacuo. Afterwards the white residue was washed with n\u2010hexane (3\u2009\u00d7\u20092 mL) to yield 4b as a white solid. Crystals suitable for single\u2010crystal X\u2010ray diffraction were obtained from a DCM solution layered with n\u2010hexane at room temperature. M.p. 150 \u00b0C. 1H NMR (CDCl3): \u03b4 = 2.22\u20132.09 , 1.94\u20131.74 , 1.70\u20131.60 , 1.54\u20131.22 ppm . 13C{1H} NMR (CDCl3): \u03b4 = 31.0\u201330.2 , 29.1 (s), 28.4\u201327.7 , 27.3 (s), 27.1\u201326.7 , 26.5\u201325.8 ppm . 31P{1H} NMR (CDCl3): \u03b4 = \u20135.7 ppm (s). IR: \u03bd\u0303 = 2931 , 2852 , 1447 , 1400 , 1113 (w), 846 cm\u20131 (w). MS (ESI(+), chloroform/MeOH): m/z = 839.1 [Ag2L2Br]+, 381.1 [AgL]+; L = 1b. C15H30AgBrP2: calcd. C 39.16, H 6.57\u2009%; found C 39.23, H 6.65\u2009%.Synthesis of Trisdisilver(I) Dibromide (5): AgBr was added as a solid to a stirred solution of 1a in 10 mL DCM. The Schlenk flask was immediately wrapped in aluminum foil and the reaction mixture stirred at room temperature for 4 h, filtered and all volatiles were removed in vacuo. Afterwards the white residue was washed with n\u2010hexane (3\u2009\u00d7\u20092 mL) to yield 5 as a white solid. Crystals suitable for single\u2010crystal X\u2010ray diffraction were obtained from a DCM/toluene mixture layered with n\u2010hexane at room temperature. M.p. 139 \u00b0C. 1H NMR (CDCl3): \u03b4 = 2.18\u20132.05 , 1.94\u20131.81 , 1.79\u20131.68 , 1.60\u20131.48 , 1.45\u20131.38 ppm . 13C{1H} NMR (CDCl3): \u03b4 = 34.0\u201333.4 , 29.3 (s), 27.9 , 27.4 (s), 27.0 ppm . 31P{1H} NMR (CDCl3): \u03b4 = \u20139.1 ppm (s). IR: \u03bd\u0303 = 2921 , 2855 , 1447 , 1409 , 1109 (w), 1069 (w), 848 cm\u20131 (w). MS (ESI(+), DCM/MeOH): m/z = 783.0 [Ag2L2Br]+, 595.2 [AgL2]+; L = 1a. C39H78Ag2Br2P6: calcd. C 42.26, H 7.09\u2009%; found C 42.28, H 7.04\u2009%.Crystal Structure Determinations: The data were collected on a Gemini diffractometer (Rigaku Oxford Diffraction) using Mo\u2010K\u03b1 radiation (\u03bb = 71.073 pm) and \u03c9\u2010scan rotation. Data reduction was performed with CrysAlisPro2: SHELXS\u201097;4a: SIR\u2010923, 4b, 5: SHELXT\u201020144a, all structures are disordered to a certain extent. Mostly the five\u2010membered C4H8P ring was disordered, and for 3 and 4b some of the CnH2n chains were disordered as well. Details concerning data collection and crystallographic details are summarized in Table 2), CCDC\u20101974714 (3), CCDC\u20101974716 (4a), CCDC\u20101974717 (4b), and CCDC\u20101974715 (5) .Crystallographic data for the structures in this paper have been deposited with the Cambridge Crystallographic Data Centre, CCDC, 12 Union Road, Cambridge CB21EZ, UK. Copies of the data can be obtained free of charge on quoting the depository numbers CCDC\u20101974713 (Supporting Information (see footnote on the first page of this article): 1H, 13C{1H}, 31P{1H} NMR, IR and mass spectra (ESI(+)) of complexes 2, 3, 4a, 4b and 5, and 31P{1H} NMR spectrum of the reaction mixture of 1a with AgBr, ratio 3:2.https://doi.org/10.1002/zaac.202000001 or from the author.Supporting information for this article is available on the WWW under Supporting InformationClick here for additional data file."} +{"text": "A symmetrical dicarbonohydrazide was used to synthesize a centrosymmetric tetra\u00adnuclear zinc(II) complex in which two of the zinc cations are penta\u00adcoordinated and the other two are hexa\u00adcoordinated. II complex, [Zn4(C13H11N6O)2Cl6(H2O)2] or {[Zn2(HL)(H2O)(Cl2)](\u03bcCl)2[Zn2(HL)(H2O)(Cl)]}2, was synthesized by mixing an equimolar amount of a methanol solution containing ZnCl2 and a methanol solution containing the ligand H2L . In the tetra\u00adnuclear complex, each of the two ligand mol\u00adecules forms a dinuclear unit that is connected to another dinuclear unit by two bridging chloride anions. In each dinuclear unit, one ZnII cation is penta\u00adcoordinated in a N2OCl2 in a distorted square-pyramidal geometry, while the other ZnII cation is hexa\u00adcoordinated in a N3OCl2 environment with a distorted octa\u00adhedral geometry. The basal plane around the penta\u00adcoordinated ZnII cation is formed by one chloride anion, one oxygen atom, one imino nitro\u00adgen atom and one pyridine nitro\u00adgen atom with the apical position occupied by a chloride anion. The basal plane of the hexa\u00adcoordinated ZnII cation is formed by one chloride anion, one hydrazinyl nitro\u00adgen atom, one imino nitro\u00adgen atom and one pyridine nitro\u00adgen atom with the apical positions occupied by a water oxygen atom and a bridged chloro anion from another dinuclear unit, leading to a tetra\u00adnuclear complex. A series of intra\u00admolecular C\u2014H\u22efCl hydrogen bonds is observed in each tetra\u00adnuclear unit. In the crystal, the tetra\u00adnuclear units are connected by inter\u00admolecular C\u2014H\u22efCl, C\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming a planar two-dimensional structure in the ac plane.A tetra\u00adnuclear Zn S-cis or S-trans, yielding different structures with the same metal cation. These ligands can coordinate to transition metals in a penta\u00addentate or hexa\u00addentate manner complex 4](NO3)4\u00b72H2O where H2L1 is 1,5-bis\u00ad[1-(pyridin-2-yl)ethyl\u00adidene)carbonohydrazide]. The study of the fluorescence properties of the ligand H2L1 and its complex revealed that complexation increased the fluorescent properties of the ligand (H2O)(Cl2)](\u03bcCl)2[Zn2(HL)(H2O)(Cl)]}2 where H2L is 1,5-bis\u00ad(pyridin-2-yl\u00admethyl\u00adene)carbono\u00adhydrazide.The behavior of these mol\u00adecules has attracted the inter\u00adest of chemists working in coordination chemistry. The free dicarbonohydrazide exhibits biological activities /60 where \u03b2 and \u03b1 are the largest values of the bond angles around the central atom]; \u03c4 = 0 for a perfect square pyramidal geometry while \u03c4 = 1 for a perfect trigonal\u2013bipyramidal geometry. In the case of the title complex, the \u03c4 value of 0.1085 is indicative of a distorted square-pyramidal geometry around the Zn1 center. The equatorial plane is occupied by atoms N5, N6, Cl3, O2 while the apical position is occupied by Cl2. The angles N5\u2014Zn1\u2014O2 [72.76\u2005(9)\u00b0], O2\u2014Zn1\u2014Cl3 [96.00\u2005(6)\u00b0], Cl3\u2014Zn1\u2014N6 [97.10\u2005(8)\u00b0] and N6\u2014Zn1\u2014N5 [75.82\u2005(10)\u00b0] deviate from those for a regular square pyramid. The transoid angles in the basal plane O2\u2014Zn1\u2014N6 and N5\u2014Zn1\u2014Cl3 deviate severely from linearity with values of 144.87\u2005(10) and 138.36\u2005(8)\u00b0, respectively \u2013111.20\u2005(7)\u00b0 and transoid angles of 171.67\u2005(8)\u00b0 [N2\u2014Zn2\u2014Cl1] and 148.24\u2005(10)\u00b0 [N4\u2014Zn2\u2014N1]. The sum of the angles subtended by the atoms in the plane is 359.77\u00b0. The apical positions are occupied by O1 and Cl1i with O 1\u2014Zn2\u2014Cl1i = 172.20\u2005(7)\u00b0 \u00b0 is in accordance with the value reported for the complex di-\u03bc-chlorido-bis\u00ad{[2-({[2-(2-pyrid\u00adyl)eth\u00adyl](2-pyridyl\u00admeth\u00adyl)amino}\u00admeth\u00adyl)-phen\u00adol]zinc(II)} bis\u00ad(perchlorate) dihydrate and 2.7489\u2005(10)\u2005\u00c5, respectively, agree with those for a chloride anion in a bridging position or diprotonated (H4L2+) and additionally one Dy complex mol\u00adecule in which HL\u2212 and L2\u2212 are present as ligands. Among the diprotonated mol\u00adecules, three different counter-ions are present: I\u2212 in AVOSOV : 3439, 3204, 3198, 3055, 2936, 1684, 1582, 1610, 1582, 1532, 1467, 1360, 1274, 1131. 1H NMR : 7.6\u20138.72 ; 10.82 ; 8.03 . 13C NMR : 157.9 (C=O); 154.70 (CPy); 148.07 (CPy); 146.67 (C=N) imine; 137.60 (CPy); 123.00 (CPy); 119.09 (CPy).Carbonohydrazide was introduced into a 100\u2005mL flask containing 20\u2005mL of methanol. To the resulting suspension was added a methano\u00adlic solution containing 2-pyridine\u00adcarbaldehyde (4.757\u2005g 44.4\u2005mmol) and two drops of glacial acetic acid. The mixture was stirred under reflux for 2\u2005h. After being kept for two days at 277\u2005K, the resulting orange solution yielded a precipitate, which was recovered by filtration. The solid was washed successively with cold methanol (2 \u00d7 10\u2005mL) and diethyl ether (2 \u00d7 10\u2005mL) before being dried under PSynthesis of the title complex2L in 10\u2005mL of methanol and a methano\u00adlic solution of ZnCl2 . A yellow solution was obtained after stirring for 1\u2005h at room temperature. The solution was filtered, and the filtrate left for slow evaporation. After two weeks, yellow crystals suitable for X-ray diffraction were collected, yield 87.9%. Analysis calculated for [C26H26Cl6Zn4N12O4] C, 29.89; H, 2.51; N, 16.09. Found: C, 29.88; H, 2.49; N, 16.05. \u039bM (S cm2 mol\u2212): 11. IR (cm\u22121): 3428, 3116, 3043, 1585, 1553, 1497, 1461, 1377, 1313, 1226, 1143, 820.The title complex was prepared by mixing a solution of HUiso(H) = 1.2Ueq(N) or 1.5Ueq(O). C atoms were placed in calculated positions and refined as riding with C\u2014H = 0.93\u2005\u00c5 and Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989020009834/ex2035sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020009834/ex2035Isup2.hklStructure factors: contains datablock(s) I. DOI: 2011426CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-019-45003-7, published online 14 June 2019Correction to: The Supplementary Information file originally published with this Article contained errors.In the \u2018Detailed experimental procedures:\u2019 section,\u201c106.57 (c-2\u2019)\u201dnow reads:\u201c10657 (C-2\u2019)\u201dand\u201cmp 91\u201392\u2009\u00b0C\u201dnow reads:\u201cm.p. mp 91\u201392\u2009\u00b0C\u201d.Finally, the original file contained tracked changes.These errors have been corrected in the Supplementary Information that now accompanies the Article."} +{"text": "To the Editor,Colorectal cancer (CRC) is a malignance ranking the third cause of death in malignant tumors.P value\u00a0=\u00a07.458 \u00d7 10\u221218) and indicated a favorable prognosis in patients with CRC (P\u00a0=\u00a0.016), which was inconsistent with our general understanding in other type of cancers , \u22120.71 (P\u00a0=\u00a08 \u00d7 10\u221257), and 0.71 (P\u00a0=\u00a01 \u00d7 10\u221255), respectively showed that CCDC80 expression level was independent predictive factor Figure\u00a0. Kaplan\u2010) Figure\u00a0. The RNA) Figure\u00a0.To explore the relationship between the drugs sensitivity and CCDC80 expression, the Genomics of Drug Sensitivity in Cancer database was utilized to analyze the correlation coefficient of CCDC80 level and IC50 of multiple drugs Figure\u00a0. To our Next, we overexpressed CCDC80 in Lovo cells by lentivirus infection and validated by RT\u2010PCR Figure\u00a0. ConsistP\u00a0=\u00a05.32 \u00d7 10\u221202), dendritic cells (P\u00a0=\u00a07.04 \u00d7 10\u221240), macrophage cells (P\u00a0=\u00a03.52 \u00d7 10\u221256), CD4+ T cells (P\u00a0=\u00a08.64 \u00d7 10\u221229), CD8+ T cells (P\u00a0=\u00a04.14\u00a0\u00d7 10\u221208), and neutrophil cells (P\u00a0=\u00a03.34 \u00d7 10\u221228) Figure\u00a0. Furthern Figure\u00a0. Among tThe authors declare no conflict of interest.Guangci Distinguished Young Scholars Training Program; Grant Number: GCQN\u20102019\u2010B17.Yu\u2010Jun Dai, Da\u2010Wei Wang, and Ling\u2010Ling Shu designed the concept and experiments. Wei\u2010Da Wang, Kun\u2010Hao Bai, and Guo\u2010Yan Wu performed the experiments; Wei\u2010Da Wang, Si\u2010Yuan He, Xin Huang, Qian\u2010Yi Zhang, Pei\u2010Dong Chi, and Liang Li collected the data and did the analysis. Wei\u2010Da Wang, Yu\u2010Jun Dai, and Guo\u2010Yan Wu prepared the manuscript draft. Yu\u2010Jun Dai and Da\u2010Wei Wang provided research support and revised the manuscript. All the authors approved the final proof.figureS1Click here for additional data file.figureS2Click here for additional data file.tableS1Click here for additional data file.tableS2Click here for additional data file.tableS3Click here for additional data file.tableS4Click here for additional data file.Supporting informationClick here for additional data file."} +{"text": "The structure is composed of cation\u2013oxygen bilayers, which are surrounded by hydro\u00adphobic methyl\u00adethyl chains on both sides. Stacking of these sandwiches forms the structure. The potassium cations are situated in an irregular coordination polyhedron composed of seven O atoms.The structure of poly[bis\u00ad(\u03bc 2(C4H7O2)2(H2O)]n, is composed of stacked sandwiches, which are formed by cation\u2013oxygen bilayers surrounded by methyl\u00adethyl hydro\u00adphobic chains. These sandwiches are held together by van der Waals inter\u00adactions between the methyl\u00adethyl groups. The methyl\u00adethyl groups are disordered over two positions with occupancies 0.801\u2005(3):0.199\u2005(3). The potassium cations are coordinated by seven O atoms, which form an irregular polyhedron. There is a water mol\u00adecule, the oxygen atom of which is situated in a special position on a twofold axis (Wyckoff position 4e). The water H atoms are involved in Owater\u2014H\u22efOcarbox\u00adyl hydrogen bonds of moderate strength. These hydrogen bonds are situated within the cation\u2013oxygen, i.e. hydro\u00adphilic, bilayer.The structure of the title compound, [K These sandwiches are bonded by van der Waals forces.In all of these structural types, water mol\u00adecules can occur; examples are given in Table\u00a01M+CnHn+12COO\u2212, n > 2, as follows from the known structures of Li(C3H5O2) , catena-[(\u03bc2-propano\u00adato)thallium(I)(propano\u00adato)thallium(I)] have been reviewed. The motif of stacked layers, however, seems to be typical for simple alkali alkanoates Thus, the typical motif of separated hydro\u00adphobic and hydro\u00adphilic parts of the mol\u00adecules can be generalized for carboxyl\u00adates other than formates and acetates.nHn+12COO\u2212, n > 2, hinder possible applications of these compounds, although there are some exceptions such as lanthanide zinc butyrates or their analogues, which have been applied for the synthesis of lanthanide\u2013zinc\u2013oxygen nanoparticles 1\u00a0\u2212\u00a0x, y, z; (ii) x, \u2212y, z; (iii) x, y, z; (iv) x, \u22121\u00a0+\u00a0y, z.) It is also worth mentioning that the distances between the cation and the oxygen atoms belonging to the same carboxyl\u00adate are quite different: K1\u2014O1 = 3.1113\u2005(13)\u2005\u00c5 and K1\u2014O2 = 2.8056\u2005(13)\u2005\u00c5.The structural unit of the title compound is shown in Fig.\u00a01The prominent feature of the title structure is the presence of an oxygen\u2013metal bilayer, which is surrounded by methyl\u00adethyl chains on both sides Fig.\u00a02. This biwater\u2014H\u22efOcarboxyl\u00adate hydrogen bonds of moderate strength and 3.01\u2005(2)\u2005\u00c5, respectively, see Fig.\u00a03v, the bond valence of which is 0.0385\u2005(1) 6 = 105.00\u2005(1)\u00b0 while Uiso(H1O3) = 1.5Ueq(O3). A trial refinement showed that the water oxygen was fully occupied. The C1\u2014C2, C1\u2014C2a bonds were restrained to be equal [1.540\u2005(1)\u2005\u00c5] as were C2\u2014C3, C2\u2014C3a and C2\u2014C4, C2\u2014C4a [1.500\u2005(1)\u2005\u00c5]. These values were found to yield the lowest R factors. Moreover, angle restraints to C3\u2014C2\u2014C4 and C3a\u2014C2a\u2014C4a were also applied. Of course, these C\u2014C distances are affected by a large thermal agitation and are less reliable, as are the geometric parameters, compared to those of atom C1.The methane\u00adtriyl hydrogens H110.1107/S2056989020012591/dj2009sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989020012591/dj2009Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020012591/dj2009Isup3.smiSupporting information file. DOI: 2032320CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "We performed batch incubation experiments with an anammox-dominated biomass to investigate nitrogen (N) and oxygen (O) isotopic fractionation factors during anammox and also examined apparent isotope fractionation factors during anammox in an actual wastewater treatment plant. We conducted one incubation experiment with high \u03b418O of water to investigate the effects of water \u03b418O. The N isotopic fractionation factors estimated from incubation experiments and the wastewater treatment plant were similar to previous values. We also found that the N isotopic effect (15\u03b5NXR of \u201377.8 to \u201365.9\u2030 and 15\u0394NXR of \u201331.3 to \u201330.4\u2030) and possibly O isotopic effect (18\u03b5NXR of \u201320.6\u2030) for anaerobic nitrite oxidation to nitrate were inverse. We applied the estimated isotopic fractionation factors to the ordinary differential equation model to clarify whether anammox induces deviations in the \u03b418O vs \u03b415N of nitrate from a linear trajectory of 1, similar to heterotrophic denitrification. Although this deviation has been attributed to nitrite oxidation, the O isotopic fractionation factor for anammox is crucial for obtaining a more detailed understanding of the mechanisms controlling this deviation. In our model, anammox induced the trajectory of the \u03b418O vs \u03b415N of nitrate during denitrification to less than one, which strongly indicates that this deviation is evidence of nitrite oxidation by anammox under denitrifying conditions.Isotopic fractionation factors against However, they lacked information on the 18O fractionation factor for anammox, and assumed that the 18O fractionation factor during NO3\u2013 production by anammox was similar to that for aerobic nitrite oxidation to NO3\u2013 by nitrifiers (nitrification). Thus, it is essential to investigate the 15N and 18O fractionation factors during anammox not only for the better use of the \u03b418O and \u03b415N of NO3\u2013 in anammox studies, but also to obtain a more detailed understanding of 15N and 18O fractionation.Studies on 18O fractionation factors during anammox. We calculated apparent 15N and 18O fractionation factors with data collected from a wastewater treatment plant (WWTP) at which anammox reactors were installed at the final stage of treatment .In the first experiment (Experiment A), the biomass in the reactor was sampled and incubated with the media used for the reactor, while the sampled biomass was re-suspended in fresh, chemically defined media in the second (Experiment B) and third (Experiment C) experiments. Difficulties were associated with performing incubation experiments with the anammox biomass for isotopic measurements, and, thus, we employed slightly different settings and operations to facilitate constant and active anammox reactions. In each experiment, 15\u200d \u200dmL of the biomass suspension in media solution was filtered with filter paper and differences in filter weights before and after filtration were used to calculate the suspended solid (S.S.) concentration after the filter had been oven-dried (at 105\u00b0C).2 gas to remove dissolved oxygen (DO). pH, DO, and the concentrations of NH4+ and NO2\u2013 were regularly measured to confirm the anammox activity of the biofilm, and pH (8.0) was maintained by adding KH2PO4 and Na2HPO4\u00b712H2O solution. After the addition of NaNO2, (NH4)2SO4, and NaHCO3, we started the incubation and sampled 10\u200d \u200dmL of media. Sampled media were filtered with a 0.20-\u03bcm syringe filter and then split into three; one for NO3\u2013 followed by the removal of NO2\u2013 (2\u2013 with high pH by the addition of 2M 2\u2013 andNaOH solution to prevent oxygen atom exchange between NO water (4+ with low pH by the addition of 4.8 M H2SO4 to prevent NH4+ from volatilizing. These subsamples were frozen (\u201330\u00b0C) until further analyses.The biofilm and incubation media solution were sampled from the incubation membrane in the anammox reactor (SI Text 1.1). The incubation was performed anaerobically in the glovebox at room temperature (25\u201330\u00b0C) after the purging of media by N of NO2\u2013 , another2 purged) in the glovebox. Media consisted of NaHCO3 502\u200d \u200dmg L\u20131; MgSO4\u00b77H2O 603\u200d \u200dmg L\u20131; CaCl2 180.5\u200d \u200dmg L\u20131; KH2PO4 169\u200d \u200dmg L\u20131; Na2HPO4\u00b712H2O 282\u200d \u200dmg L\u20131; trace elements of solution I 0.5\u200d \u200dmL and solution II 0.5\u200d \u200dmL. We added NaNO2 and (NH4)2SO4 to media (500\u200d \u200dmL) with the anammox granules and then started the incubation at room temperature (25\u201330\u00b0C). We monitored pH (7.9 to 8.8) and NO2\u2013 to assess the progress of anammox. Sampling was performed as described in Experiment A.The granule biomass that accumulated at the bottom of the anammox reactor was sampled. Granules were rinsed anaerobically with new media (N\u03b418O of water (\u03b418OH2O) was markedly higher (229\u2030) than that in Experiments A and B (\u20138\u2030). This \u201cheavy\u201d water was prepared by mixing 18O-labeled water (10% atom 18O) with Milli-Q water. During the incubation, media with biofilms were shaken at a constant temperature (30\u00b0C) and continuously purged with a gas mixture (95% Ar\u00a0+\u00a05% CO2) to maintain low DO levels. pH ranged between 7.1 and 7.5 and the monitoring and sampling scheme was identical to Experiment BWe incubated the biofilm collected from the incubation membrane in the anammox reactor with the same media used in Experiment B; however, the 4+ concentrations were measured during the incubation by the o-phthaldialdehyde (OPA) method with USGS 32, 34, 35, and IAEA-2 as standards, and with the azide method with TUAT-NO2-1 to TUAT-NO2-5 were evaluated using GC-IRMS with the denitrifier method after the conversion of NH4+ to NO3\u2013 by persulfate oxidation \u20131 and (R_SampleO/R_Oxygen)\u20131 where R_SampleN and R_SampleO are [15N/14N] and [18O/16O] of the sample, respectively, R_Nitrogen is [15N/14N] of atmospheric N2 and R_Oxygen is [18O/16O] of Vienna Standard Mean Ocean Water , residual NH4+ (\u03b415NNH4+_ANX), and the fraction of NH4+ reacting (NH4+f) in ANX reactor (15\u0394AMX (The ammonium oxidation to N reactor at a ste15\u0394AMX = (\u03b415NNH4+_ANX \u2013 \u03b415NNH4+_NT) / NH4+f --- eq. (1)NH4+f = ([NH4+]NT \u2013 [NH4+]ANX) / [NH4+]NT4+]NT and [NH4+]ANX are the NH4+ concentrations in NT and ANX reactors, respectively.where ANX / ([NO2 \u2013 ]NT \u2013 [NO2\u2013]ANX)b = ([NO3\u2013]ANX \u2013 [NO3\u2013]NT) / ([NO2\u2013]NT \u2013 [NO2\u2013]ANX)\u03b415NNO2\u2013_NT is \u03b415NNO2\u2013 in NT reactor and the concentrations of NO2\u2013 and NO3\u2013 in NT and ANX reactors are [NO2\u2013]NT, [NO2\u2013]ANX, [NO3\u2013]NT, and [NO3\u2013]ANX, respectively.where The combination of eqs. (2) and (4) gives\u03b415NNO2\u2013_NT = (1 \u2013 a \u2013 b) \u00d7 \u03b415NN2_ANX + a \u00d7 \u03b415NNO2\u2013_ANX + b\u00d7 \u03b415NNO3\u2013_ANXa \u2013 b) \u00d7 (\u03b415NNO2\u2013_ANX \u2013 15\u0394AMXNIR) + a\u00d7 \u03b415NNO2\u2013_ANX + b \u00d7 \u03b415NNO3\u2013_ANX= (1 \u2013 \u03b415NNO2\u2013_ANX \u2013 15\u0394AMXNIR \u2013 a \u00d7 \u03b415NNO2\u2013_ANX+ a \u00d7 15\u0394AMXNIR \u2013 b \u00d7 \u03b415NNO2\u2013_ANX + b \u00d7 15\u0394AMXNIR+ a \u00d7 \u03b415NNO2\u2013_ANX + b \u00d7 \u03b415NNO3\u2013_ANX= b \u00d7 (\u03b415NNO3\u2013_ANX \u2013 \u03b415NNO2\u2013_ANX) + \u03b415NNO2\u2013_ANX+ 15\u0394AMXNIR \u00d7 (a + b \u2013 1)= b \u00d7 15\u0394NXR) + \u03b415NNO2\u2013_ANX + 15\u0394AMXNIR \u00d7 (a + b \u2013 1)= \u2013 (15\u0394AMXNIR = [\u03b415NNO2\u2013_ANX \u2013 \u03b415NNO2\u2013_NT \u2013 b \u00d7 15\u0394NXR] / (a + b \u2013 1) --- eq. (5)18O fractionation during nitrite oxidation to nitrate, we followed the approach described by 18EAMXcombined) because of the lack of detailed information on isotopic fractionation for the nitrite oxidation and oxygen atom incorporation during nitrite oxidation. Thus, we calculated 18EAMXcombined as follows:To calculate 18EAMXcombined = 2/3 \u03b418ONO2\u2013_ANX + 1/3 \u03b418OH2O \u2013 \u03b418ONO3\u2013_ANX--- eq. (6)\u03b418ONO2\u2013_ANX, \u03b418OH2O, and \u03b418ONO3\u2013_ANX are the 18O ratios of NO2\u2013, water, and NO3\u2013 in ANX reactor, respectively.where 2\u2013 were then estimated from isotopic data.We developed an ordinary differential equation model as described by Fluxes regarding the anammox process are defikAMO14N \u00d7 [14NH4+] --- eq. (7)AMX = AMXNIR = x / (1 \u2013 x)) --- eq. (8)NXR = AMXNIR \u00d7 (14N) fluxes of ammonium oxidation, nitrite oxidation, and reduction by anammox .where AMX, NXR, and AMXNIR are the in the a [NO2\u2013]) and the 2\u2013;Regarding NO14NO2\u2013] = \u2013 NXR \u2013 AMXNIR --- eq. (9)d/dt [15NO2\u2013] = \u2013 (R_NitriteN \u00d7 NXR / 15\u03b5NXR)15\u03b5AMXNIR) --- eq. (10)\u2013 (R_NitriteN \u00d7 AMXNIR / d/dt [16O2\u2013] = \u2013 2NXR \u2013 2AMXNIR --- eq. (11)d/dt [N16O18O\u2013] = \u2013 (R_NitriteO \u00d7 2NXR / 18\u03b5NXR)\u2013 (R_NitriteO \u00d7 2AMXNIR / 18\u03b5AMXNIR)\u2013 N16O18O\u2013exch_OUT + N16O18O\u2013exch_INd/dt [N18\u03b5NXR)\u2013 (R_NitriteO \u00d7 2AMXNIR / 18\u03b5AMXNIR)\u2013 kexch \u00d7 [N16O18O\u2013] + kexch\u00d7R_WaterO / 18\u03b5EQ--- eq. (12)= \u2013 , N16O18O\u2013exch_OUT, and N16O18O\u2013exch_IN as described by 2\u2013 and H2O.where R_NitriteO, R_NitrateO, R_NitriteN, and R_NitrateN are the e and pH . We appl3\u2013;Regarding NO14NO3\u2013] = NXR --- eq. (13)d/dt [15NO3\u2013] = (R_NitriteN \u00d7 NXR / 15\u03b5NXR) --- eq. (14)d/dt [16O3\u2013] = 3 NXR --- eq. (15)d/dt [N18O16O2\u2013] = (R_NitriteO \u00d7 2 NXR / 18\u03b5NXR)+ (R_WaterO \u00d7 NXR) / 18\u03b5H2ONXR) --- eq. (16)d/dt [N18\u03b5H2ONXR d/dt [15NH4+] = \u2013 (R_AmmoniumN \u00d7 AMX/ 15\u03b5AMX) --- eq. (18)d/dt [15N / 14N of [NH4+] and 15\u03b5AMX is the N isotopic fractionation factor for NH4+ consumption by anammox (where R_AmmoniumN is the anammox .2\u2013 and NH4+ to N2 and NO3\u2013 is as follows (The approximate stoichiometry of the anammox process converting NO2\u2013 + 1NH4+ \u2192 1N2 + 0.3NO3\u2013 + 2H2O --- eq. (19)1.3NOx) together with kAMO14N with concentration data, which provided the AMX, AMXNIR, and NXR fluxes used in the calculation above . We assigned the range from 0 to 60\u2030 to estimate isotopic fractionation factors. We considered this 60\u2030 range for the curve-fitting estimate to be reasonable because isotopic fractionation factors larger than 60\u2030 are rarely observed and consequently 18EAMXcombined , were not all successfully estimated for Experiments B and C. Based on these uncertainties in parameter estimations, we did not report these calculated values for Experiments B and C; however, we speculate that these calculated parameter sets support 18\u03b5AMXNIR as normal and 18\u03b5NXR being inverse isotope fractionation, as discussed below for Experiment A. The curve-fitting function (\u201cmultiple-fit\u201d in BM software) (\u20136. BM codes for the anammox model for curve fittings with concentrations and isotopic data are provided in the Zenodo website (https://doi.org/10.5281/zenodo.3895346) and However, this stoichiometry between nitrite removal and nitrate production has been reported to vary d/dt [15NO2\u2013] = \u2013 (R_NitriteN \u00d7 NXR / 15\u03b5NXR)+ (R_NitrateN \u00d7 NAR / 1518\u03b5NAR)\u2013 (R_NitriteN \u00d7 DENNIR / 15\u03b5DENNIR)\u2013 (R_NitriteN \u00d7 AMXNIR / 15\u03b5AMXNIR) --- eq. (21)d/dt [16O2\u2013] = \u2013 2 NXR + 2 NAR \u2013 2 DENNIR \u2013 2 AMXNIR --- eq. (22)d/dt [N1518\u03b5NAR + (R_NitrateO \u00d7 2 NAR / 1518\u03b5NAR) / 18\u03b5H2OBRNAR\u2013 (R_NitriteO \u00d7 2 DENNIR / 18\u03b5DENNIR)\u2013 (R_NitriteO \u00d7 2 AMXNIR / 18\u03b5AMXNIR) --- eq. (23a)d/dt [N18\u03b5H2OBRNAR is the 18O fractionation factor for the \u201cbranching effect\u201d + 1]d/dt = NXR \u2013 NAR --- eq. (24)d/dt [15NO3\u2013] = (R_NitriteN \u00d7 NXR / 15\u03b5NXR) \u2013 (R_NitrateN \u00d7 NAR / 1518\u03b5NAR) --- eq. (25)d/dt [18O16O2\u2013] = 3 NXR \u2013 3 NAR --- eq. (26)d/dt [N18O16O2\u2013] = (R_NitriteO \u00d7 2 NXR / 18\u03b5NXR)+ (R_WaterO \u00d7 NXR / 18\u03b5H2ONXR)\u2013 (R_NitrateO \u00d7 3 NAR / 1518\u03b5NAR) --- eq. (27)d/dt and [NO2\u2013] decreased during anammox (\u03b418ONO2\u2013 and \u03b418ONO3\u2013 did not change in Experiment A (\u03b418ONO3\u2013 increased by ~2\u2030 and \u03b418ONO2\u2013 decreased by ~3\u2030 in Experiment B (\u03b418O of H2O (229\u2030), \u03b418ONO2\u2013 and \u03b418ONO3\u2013 rapidly increased for Kuenenia stuttgartiensis in batch incubation experiments (Candidatus Jettenia\u201d with the nirK gene ( 2 \u00b1 2\u2030) . The simirK gene .15N and 18O fractionation factors during anaerobic nitrite oxidization to NO3\u2013 of \u201377.8\u2030 for 15\u03b5NXR and \u201320.6\u2030 for 18\u03b5NXR ; defined as \u2013 (18\u03b5 / 15\u03b5), where 18\u03b5 / 15\u03b5 is the ratio of isotopic fractionation for O and N during denitrification, respectively, and assigned as 1; see the inset in 2\u2013 was consumed. In the case of denitrification in which AMX / NAR is equal to 0 (indicating no anammox), \u03b418ONO3\u2013 vs \u03b415NNO3\u2013 was set to show a slope of 1 . To elucidate the relationship between \u0394, AMX / NAR, and isotopic fractionation factors, we simulated the \u03b418ONO3\u2013 and \u03b415NNO3\u2013 trajectories along with the different AMX / NAR ratios and 15\u03b5AMXNXR ; however, \u0394 depended on AMX / NAR, 15\u03b5AMXNXR, and 15\u03b5NXR , our simulation exercise revealed that \u0394, the offset from the 1:1 relationship between \u03b415N and \u03b418O, may be useful for detecting NXR (nitrite oxidation) in denitrifying systems in both freshwater and seawater.We developed an anammox-denitrification model with the estimated isotopic fractionation factors denitrification with auxiliary Nap NO3\u2013 reductase may exhibit a 2:1 rather than 1:1 relationship between \u03b415N and \u03b418O, resulting in an offset from the 1:1 relationship is not considered to be a major environmental sink for NO3\u2013 may induce an offset from the denitrification trajectory . In practice, \u0394 may be evaluated with time-course samplings or short incubation studies to investigate the occurrence of anammox, similar to denitrification. This technique will be advantageous because of its potential in evaluations of the quantitative contribution in situ of anammox versus denitrification. Although the detection and quantification of functional genes in denitrification and anammox may be readily performed, difficulties are associated with detecting the in situ occurrence of denitrification and anammox. Although the isotopic fractionation factors used also need to be considered, \u0394 is a promising parameter to complement molecular data and the results from laboratory incubation experiments in the study of anammox.We estimated Microbes Environ 35: ME20031.Kotajima, S., Koba, K., Ikeda, D., Terada, A., Isaka, K., Nishina, K., et al. (2020) Nitrogen and Oxygen Isotope Signatures of Nitrogen Compounds during Anammox in the Laboratory and a Wastewater Treatment Plant. https://doi.org/10.1264/jsme2.ME20031Supplementary Material"} +{"text": "Hypercalcemia is a frequently encountered electrolyte abnormality with a well-described differential diagnosis and classic algorithm for evaluation. The treatment for hypercalcemia is dependent on the underlying etiology. Hypervitaminosis D is an uncommon cause of hypercalcemia, but the use of vitamin D supplementation has expanded and case reports of supplemental vitamin D induced hypercalcemia have become more frequent. We present a case of hypervitaminosis D-induced altered mental status where diagnosis was delayed and additional invasive testing was performed due to an assumption regarding phosphatemia. Vitamin D is a steroid hormone crucial for both bone health and calcium homeostasis . It can 2 95%. Physical exam revealed bilateral upper extremity tremors and obtundation. Pupils were symmetric and reactive to light, and he periodically tracked with his eyes. Neurological exam, including reflexes, was limited due to altered mental status. Cardiopulmonary exam was benign. Chest X-ray was notable for a normal cardiopulmonary silhouette. Electrocardiogram revealed normal sinus rhythm with a QTc interval of 409\u00a0ms. Computed tomography of the brain without contrast showed normal gray-white matter differentiation. A 64-year-old man with a history of chronic obstructive pulmonary disease and ethanol abuse use presented to the emergency department with altered mentation. He was obtunded on arrival, unable to provide any additional history. Presenting vitals were notable for blood pressure of 150/93, pulse of 69 beats per minute, temperature of 97.3\u00a0\u00b0F (36.3\u00a0\u00b0C), respiratory rate of 18 BPM, and SpOLaboratory evaluation revealed a sodium 143\u00a0mEq/L (135\u00a0\u2013\u00a0145\u00a0mEq/L), potassium 3.0\u00a0mEq/L (3.6\u00a0\u2013\u00a05.1\u00a0mEq/L), chloride 105 mEq/L (98\u00a0\u2013\u00a0110\u00a0mEq/L), bicarbonate 27\u00a0mEq/L (22\u00a0\u2013\u00a032 mEq/L), blood urea nitrogen 14\u00a0mg/dL (6\u00a0\u2013\u00a024\u00a0mg/dL), creatinine 1.11\u00a0mg/dL (0.64\u00a0\u2013\u00a01.27 mg/dL), undetectable blood ethanol level, blood glucose 95\u00a0mg/dL (67\u00a0\u2013\u00a099\u00a0mg/dL), calcium 12.8\u00a0mg/dL (8.5\u00a0\u2013\u00a010.5 mg/dL), ionized calcium 6.3\u00a0mg/dL (4.2\u00a0\u2013\u00a05.2\u00a0mg/dL), phosphorus of 2.2\u00a0mg/dL (2.4\u00a0\u2013\u00a04.8 mg/dL), and albumin 4.0\u00a0g/dL (3.5\u00a0\u2013\u00a05.0\u00a0g/dL). Initial management consisted of volume expansion with 3\u00a0L normal saline without improvement in mentation. Calcitonin was given on hospital day\u00a02 and zoledronic acid was given on hospital day\u00a03, but hypercalcemia persisted. Further labwork returned with intact parathyroid hormone (iPTH) 17\u00a0pg/mL (18\u00a0\u2013\u00a080\u00a0pg/mL), parathyroid hormone related peptide (PTHrP) 9\u00a0pg/mL (pg/mL 14\u00a0\u2013\u00a027), serum protein electrophoresis (SPEP) and immunofixation (IFE) without evidence of monoclonal protein, \u03ba light chains 85.4\u00a0mg/L (3.3\u00a0\u2013\u00a019.4 mg/L), \u03bb light chains 40.8 mg/L (5.7\u00a0\u2013\u00a026.3\u00a0mg/L), and \u03b2-2 microglobulin 3.75\u00a0mg/L (0.97\u00a0\u2013\u00a02.64\u00a0mg/L). The elevation of \u03ba/\u03bb ratio in the setting of a normal glomerular filtration rate prompted evaluation for a monoclonal gammopathy. Skeletal survey was negative for lytic lesions and bone marrow biopsy was negative for plasma cell dyscrasia. Finally, the following labs resulted: 25-hydroxy vitamin D >\u00a0150\u00a0ng/mL (30.0\u00a0\u2013\u00a0100.0\u00a0ng/mL), 1,25-hydroxy vitamin D 36 pg/mL (18\u00a0\u2013\u00a072\u00a0pg/mL). The patient remained agitated and confused for the first 10\u00a0days of hospitalization, but mentation improved thereafter with calcium normalizing after 18\u00a0days. Upon recovery, he reported misunderstanding medication instructions and had been consuming 4 vitamin\u00a0D 50,000 IU tablets daily. This case highlights the risks of pathophysiologic assumptions. The findings of hypercalcemia with appropriately suppressed iPTH and low PTHrP resulted in an evaluation for hypervitaminosis D. However, hypophosphatemia directed the evaluation away from hypervitaminosis D, leading to additional evaluation for paraproteinemia. The finding of an abnormal \u03ba-to-\u03bb ratio and elevated \u03b2-2 microglobulin, resulted in evaluation for plasma cell dyscrasia with a bone marrow biopsy. The patient had an elevated vitamin D 25OHD and normal calcitriol revealing the ultimate diagnosis to be hypervitaminosis D. The presentation of hypercalcemia frequently varies with the level of serum calcium. Individuals with mild hypercalcemia (<\u00a012\u00a0mg/dL) are typically asymptomatic . ModeratHypervitaminosis D classically presents as hypercalcemia with hyperphosphatemia and low PTH , 12. VitSerum phosphorus regulation is likely incompletely understood. Phosphate is regulated by PTH, fibroblast growth factor 23 (FGF23), and 1,25OHD . PhosphaThis case highlights the importance of having a broad differential for hypercalcemia. We recommend that hypervitaminosis D be considered in patients with hypercalcemia and low PTH regardless of phosphorus level. The authors did not receive support for this work. The authors declare no conflict of interest."} +{"text": "Litylenchus crenatae mccannii (Anguinidae) nematodes by systematically testing thermostable polymerases, proofreading enzymes and buffers. The combination of thermostable DreamTaq\u2122, proofreading Pfu polymerase, and PicoMaxx\u2122 buffer provided the best results. These nematodes are the subject of surveys currently active at many sites in the northeastern United States. This new, optimized PCR protocol will be useful for diagnostic labs associated with the surveys.Generating DNA markers for microscopic plant parasitic nematodes can be especially difficult if only a few valuable, tiny specimens are available. Providing a reliable maximum amount of unambiguous genetic information from single nematodes is especially important when identifying damaging, regulated nematodes of importance to trade where a few nucleotide differences in diagnostic markers are significant. There are many possible reasons for difficulty amplifying unpurified nematode DNA for long range PCR followed by direct sequencing. Specimen age, proofreading errors and reagent compatibility during PCR are among those problems. While unsuccessful direct amplification of difficult samples may sometimes be overcome by cloning, a more expensive and time-consuming process. Therefore, long segment PCR of a large 3.5\u2009kb segment of ribosomal DNA was optimized for individual difficult-to-amplify young Fagus grandifolia) trees in Ohio, Pennsylvania, New York and Connecticut. It was discovered first in 2012 near Lake Erie in the Cleveland Metroparks, region of Ohio (Beech leaf disease (BLD) is an emerging tree disease in the Northeast for American beech . Their ribosomal DNA (rDNA) loci were amplified by PCR with the one primer set and an enhanced DNA polymerase system, and the resulting 3.5\u2009kb rDNA amplicons were directly sequenced and PicoMaxx\u2122 High Fidelity PCR System were selected and tested in this study.Commonly used o 2.9\u2009kb . It has R system . As a reLitylenchus specimens were isolated from the banding lesions of American beech leaves with BLD as described in Litylenchus and the visualization, cleanup and direct DNA sequencing, including sequencing primers, of the PCR products were performed by using the procedures described in previous studies were performed by using the procedures, including primer sets, 18S-CL-F3 and 28S-CL-R for the 3.5\u2009kb ribosomal loci, 18S-CL-F3 and 18S-CL-R7 for the 18\u2009S locus (1.7\u2009kb) and ITS-CL-F2 and 28S-CL-R for the ITS-28S (D1D2D3) loci (1.9\u2009kb) described in the previous study was carried out in a 25\u2009\u03bcl of mixture containing Platinum\u2122 Taq (10 units/\u03bcl) 0.125\u2009\u03bcl, 10X PCR Buffer Mg 2.5\u2009\u03bcl, MgCl2 (50\u2009mM) 1\u2009\u03bcl, dNTP (2.0\u2009mM each) 2.5\u2009\u03bcl, Template DNA 2\u2009\u03bcl, forward primer (10\u2009\u03bcm) 0.75\u2009\u03bcl and reverse primer (10\u2009\u03bcm) 0.75\u2009\u03bcl for either primer set 18S-CL-F3 and 28S-CL-R or ITS-CL-F2 and 28S-CL-R, and molecular biology grade water 15.375\u2009\u03bcl. The thermal cycling program was one cycle of 95\u00b0C for 3\u2009min; 36 cycles of 95\u00b0C for 30\u2009sec, 50\u00b0C for 45\u2009sec, 72\u00b0C for 3\u2009min; and final extension at 72\u00b0C for 7\u2009min.Each PCR amplification with Platinum\u2122 Taq2000\u2122 DNA Polymerase was carried out in a 25\u2009\u03bcl mixture containing Taq2000\u2122 (5 units/\u03bcl) 0.25\u2009\u03bcl, 10X PCR Buffer 2.5\u2009\u03bcl, MgCl2 (50\u2009mM) 0.25\u2009\u03bcl, dNTP (2.0\u2009mM each) 2.5\u2009\u03bcl, Template DNA 2\u2009\u03bcl, both forward primer (10\u2009\u03bcm) 0.75\u2009\u03bcl and reverse primer (10\u2009\u03bcm) 0.75\u2009\u03bcl for either primer set 18S-CL-F3 and 28S-CL-R or ITS-CL-F2 and 28S-CL-R, and molecular biology grade water 16\u2009\u03bcl. The thermal cycling program was one cycle of 95\u00b0C for 3\u2009min; 36 cycles of 95\u00b0C for 30\u2009sec, 50\u00b0C for 45\u2009sec, 72\u00b0C for 3\u2009min; and final extension at 72\u00b0C for 7\u2009min.Each PCR amplification with Taq\u2122 DNA Polymerase alone or combined with DreamTaq\u2122 was carried out in a 25\u2009\u03bcl of mixture containing TaKaRa Ex Taq\u2122 (5 units/\u03bcl) 0.125\u2009\u03bcl (or plus DreamTaq\u2122 (5 units/\u03bcl) 0.125\u2009\u03bcl), 10X Ex Taq Buffer 2.5\u2009\u03bcl, dNTP (2.5\u2009mM each) 2\u2009\u03bcl, Template DNA 2\u2009\u03bcl, forward primer 18S-CL-F3 (10 \u03bcm) 1.25\u2009\u03bcl, reverse primer 28S-CL-R (10\u2009\u03bcm) 1.25\u2009\u03bcl, and molecular biology grade water 15.875\u2009\u03bcl (or 15.75\u2009\u03bcl). The thermal cycling program was: one cycle of 98\u00b0C for 30\u2009sec; 36 cycles of 98\u00b0C for 10\u2009sec, 68\u00b0C for 5\u2009min; and final extension at 72\u00b0C for 7\u2009min.Each PCR amplification with TaKaRa Ex Taq\u2122 was carried out in a 25\u2009\u03bcl of mixture containing PicoMaxx\u2122 high fidelity PCR system (PicoMaxx\u2122 (5 units/\u03bcl)) 0.5\u2009\u03bcl (or plus DreamTaq\u2122 (5 units/\u03bcl) 0.125\u2009\u03bcl), 10\u00d7 PicoMaxx\u2122 reaction buffer (PicoMaxx\u2122 buffer) 2.5\u2009\u03bcl, dNTP (25\u2009mM each) 0.2\u2009\u03bcl, Template DNA 2\u2009\u03bcl, forward primer 18S-CL-F3 (10\u2009\u03bcm) 1.25\u2009\u03bcl, reverse primer 28S-CL-R (10\u2009\u03bcm) 1.25\u2009\u03bcl, and molecular biology grade water 17.3\u2009\u03bcl (or 17.175\u2009\u03bcl). The thermal cycling program was one cycle of 95\u00b0C for 2\u2009min; 36 cycles of 95\u00b0C for 30\u2009sec, 55\u00b0C for 45\u2009sec, 72\u00b0C for 5\u2009min; and final extension at 72\u00b0C for 7\u2009min.Each PCR amplification with PicoMaxx\u2122 High Fidelity PCR System (PicoMaxx\u2122 System) alone or combined with Dreampfu DNA polymerase alone or combined with DreamTaq\u2122 was carried out in a 25\u2009\u03bcl of mixture containing pfu (2.5 units/\u03bcl) 0.75\u2009\u03bcl (or plus DreamTaq\u2122 (5 units/\u03bcl) 0.125\u2009\u03bcl), 10\u00d7 Pfu reaction buffer, 10\u00d7 PicoMaxx\u2122 buffer, or 10\u00d7 DreamTaq\u2122 buffer 2.5\u2009\u03bcl, dNTP (25\u2009mM each) 0.2\u2009\u03bcl, Template DNA 2\u2009\u03bcl, forward primer 18S-CL-F3 (10\u2009\u03bcm) 1.25\u2009\u03bcl, reverse primer 28S-CL-R (10\u2009\u03bcm) 1.25\u2009\u03bcl, and molecular biology grade water 16.85\u2009\u03bcl (or 16.725\u2009\u03bcl). The thermal cycling program was one cycle of 95\u00b0C for 2\u2009min; 36 cycles of 95\u00b0C for 30\u2009sec, 55\u00b0C for 45\u2009sec, 72\u00b0C for 5\u2009min; and final extension at 72\u00b0C for 7\u2009min.Each PCR amplification with Pwo DNA polymerase alone or combined with DreamTaq\u2122 was carried out in a 25\u2009\u03bcl final volume consisting of two mixtures: 12.5\u2009\u03bcl of mixture A containing Pwo (5 units/\u03bcl) 0.125\u2009\u03bcl (or plus DreamTaq (5 units/\u03bcl) 0.125\u2009\u03bcl), 10\u00d7 Pwo reaction buffer or 10\u00d7 PicoMaxx\u2122 buffer 2.5\u2009\u03bcl, and molecular biology grade water 9.875\u2009\u03bcl (or 9.75\u2009\u03bcl); 12.5\u2009\u03bcl of mixture B containing dNTP (25\u2009mM each) 0.4\u2009\u03bcl, template DNA 2\u2009\u03bcl, forward primer 18S-CL-F3 (10\u2009\u03bcm) 1.25\u2009\u03bcl, reverse primer 28S-CL-R (10\u2009\u03bcm) 1.25\u2009\u03bcl. The thermal cycling program was one cycle of 95\u00b0C for 2\u2009min; 36 cycles of 95\u00b0C for 30\u2009sec, 57\u00b0C for 45\u2009sec, 72\u00b0C for 5\u2009min; and final extension at 72\u00b0C for 7\u2009min.Each PCR amplification with Each PCR amplification with Herculase\u00ae II Fusion DNA polymerase was carried out in a 25\u2009\u03bcl of mixture containing Herculase\u00ae II Fusion DNA polymerase 0.5\u2009\u03bcl, 5\u00d7 reaction buffer 5\u2009\u03bcl, dNTP (25\u2009mM each) 0.25\u2009\u03bcl, Template DNA 2\u2009\u03bcl, forward primer 18S-CL-F3 (10\u2009\u03bcm) 0.625\u2009\u03bcl, reverse primer 28S-CL-R (10\u2009\u03bcm) 0.625\u2009\u03bcl, and molecular biology grade water 16\u2009\u03bcl. The thermal cycling program was: one cycle of 95\u00b0C for 2\u2009min; 36 cycles of 95\u00b0C for 20\u2009sec, 55\u00b0C for 20\u2009sec, 72\u00b0C for 2\u2009min 15\u2009sec; and final extension at 72\u00b0C for 7\u2009min.Each PCR amplification with Phusion\u2122 High-Fidelity DNA Polymerase was carried out in a 25\u2009\u03bcl of mixture containing Phusion\u2122 High-Fidelity DNA Polymerase (2 units/\u03bcl) 0.25\u2009\u03bcl, 5\u00d7 reaction buffer 5\u2009\u03bcl, dNTP (2.5\u2009mM each) 0.5\u2009\u03bcl, Template DNA 2\u2009\u03bcl, forward primer 18S-CL-F3 (10\u2009\u03bcm) 1.25\u2009\u03bcl, reverse primer 28S-CL-R (10\u2009\u03bcm) 1.25\u2009\u03bcl, DMSO 0.25\u2009\u03bcl, and molecular biology grade water 14.5\u2009\u03bcl. The thermal cycling program was: one cycle of 95\u00b0C for 2\u2009min; 36 cycles of 95\u00b0C for 20\u2009sec, 55\u00b0C for 20\u2009sec, 72\u00b0C for 2\u2009min 15\u2009sec; and final extension at 72\u00b0C for 7\u2009min.Taq\u2122 system were carried out in 10 out of 11 Fall specimens. The direct sequencing for the three loci (3.5\u2009kb) was also conducted successfully in all specimens, except for 104H89 and 104H90 with low PCR yields that were good for sequencing only one or two loci. The 3.5\u2009kb rDNA sequences generated for the specimens, 104H82 ( MN525396) and 104H83 (MN525397) were submitted to GenBank. This result shows that DreamTaq\u2122 had the ability to amplify the 3.5\u2009kb target in most Fall specimens within the size limit by Taq DNA polymerase up to 3 to 4\u2009kb on amplicon , and ITS and 28\u2009S loci (1.9\u2009kb) within the 3.5\u2009kb target, respectively. The amplifications showed that DreamTaq\u2122 can amplify both medium 1.7\u2009kb and 1.9\u2009kb fragments with high yield seen in Both TaKaRa compare with 3B.pecimens , the Drebination . This suPwo (derived from Pyrococcus woesei), another proofreading DNA polymerase, was tested in line with the Pfu in PicoMaxx\u2122 buffer. Pwo or DreamTaq\u2122 alone . No significant amplifications of the 3.5\u2009kb target were seen in the presence of either buffer (data not shown). This confirms again that PCR buffer is another key to the success of the DreamTaq\u2122 and Pfu or Pwo combination.with Pwo . The preTaq\u00ae system and PicoMaxx\u2122 system was also performed. Taq\u00ae systems failed to amplify the 3.5\u2009kb target, but the PicoMaxx\u2122 system gave DreamTaq\u2122 dramatic leverage over the TaKaRa Ex Taq\u00ae system and ability to amplify long target (>20\u2009kb) (both Agilent and Thermo Fisher Scientific web sites). Herculase\u00ae II Fusion DNA polymerase and Phusion\u2122 High-Fidelity DNA Polymerase were tested. Fusion DNA polymerase is an engineered fusion of a proofreading polymerase and a processivity-enhancing domain and offeTaq2000\u2122, which is one of the components of the PicoMaxx\u2122 system, Platinum\u2122 Taq and DreamTaq\u2122 were also compared in Spring specimens. In the presence of their own buffers, both the long segment PCR for the 3.5\u2009kb target and the medium range PCR for the 1.9\u2009kb target were carried out. In the long segment PCR amplifications, all of the three Taqs failed to amplify the 3.5\u2009kb target and DreamTaq\u2122 can successfully amplify the 3.5\u2009kb target in the specimens where both DreamTaq\u2122 and PicoMaxx\u2122 systems failed separately.The PCR performances of b target . In the Taq-based blend systems, TaKaRa Ex Taq\u00ae DNA Polymerase in the specimen 104J58 (OH), PicoMaxx\u2122 High Fidelity PCR System in the Summer Specimen 104K17 (OH); by the combination of DreamTaq\u2122 and PicoMaxx\u2122 High Fidelity PCR System in the Summer specimens, 104K25 and 104K37 were sequenced and the resulting rDNA sequences (ITS and 28\u2009S loci) were deposited in GenBank with the accession numbers, 104H82, MN525396; 104H83, MN525397; 104J58 MN525398; 104K17, MN525399; 104k25, MN525400; 104K37, MN525401, respectively. Multiple alignments of these sequences above with the 3.5\u2009kb rDNA (MK292137 and MK292138) of the Ohio Litylenchus specimens in the previous study in these Summer specimens . DreamTaq\u2122 also demonstrated its higher sensitivity than either Taq2000\u2122 or Platinum\u2122 Taq , Pristionchus sp. (Rhabditida) and Prodorylaimus sp. (Dorylaimida) when their specimens were difficult to amplify with the one primer set and DreamTaq\u2122 (data not shown). This improvement provides high fidelity, sensitivity and yield with minimum optimization of reaction and cycling conditions. It should not be limited to long segment PCR amplification only, and could be considered for short range PCR with forensic or ancient DNA, single copy nuclear gene PCR or where improved proofreading can rescue mismatches that take place between the 3\u2019 primer termini and its target templates.Taken together, the size limit to the 3.5\u2009kb target by"} +{"text": "The title compound exhibits exceptionally weak inter\u00admolecular C\u2014H\u22ef\u03c0 hydrogen bonding of the ethynyl groups, with the corresponding H\u22ef\u03c0 separations [2.91\u2005(2) and 3.12\u2005(2)\u2005\u00c5] exceeding normal vdW distances. This bonding compliments distal contacts of the CH \u22ef\u03c0 type [H\u22ef\u03c0 = 3.12\u2005(2)\u20133.14\u2005(2)\u2005\u00c5] to sustain supra\u00admolecular layers. 14H16, exhibits exceptionally weak inter\u00admolecular C\u2014H\u22ef\u03c0 hydrogen bonding of the ethynyl groups, with the corresponding H\u22ef\u03c0 separations [2.91\u2005(2) and 3.12\u2005(2)\u2005\u00c5] exceeding normal vdW distances. This bonding complements distal contacts of the CH \u22ef\u03c0 type [H\u22ef\u03c0 = 3.12\u2005(2)\u20133.14\u2005(2)\u2005\u00c5] to sustain supra\u00admolecular layers. Hirshfeld surface analysis of the title compound suggests a relatively limited significance of the C\u22efH/H\u22efC contacts to the crystal packing (24.6%) and a major contribution from H\u22efH contacts accounting 74.9% to the entire surface.The title compound, C The shortest contacts between successive layers concern inter\u00adactions involving the methyl\u00adene groups C10H\u22efCg(C1C2)iv . The Hirshfeld surface of the mol\u00adecule, mapped over dnorm in the color range 0.0957 to 1.3378 a.u., indicates only a set of normal vdW contacts (white regions) corresponding to the closest inter\u00adactions are by far the major contributors (74.9%) to the entire surface, while the fraction of C\u22efH/H\u22efC contacts accounts for only 24.6%. The latter value may be compared with contributions of 40.0 and 32.4% calculated for \u03b1,\u03c9-octa- and deca\u00addiynes global, I. DOI: 10.1107/S2056989020005964/lh5958Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020005964/lh5958Isup3.cmlSupporting information file. DOI: 2000259CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the mol\u00adecular packing, O\u2014H\u22efO and O\u2014H\u22efS hydrogen bonds lead to supra\u00admolecular layers propagating in the ab plane.In the title tri-substituted thio\u00adurea mol\u00adecule, a substantial twist is evident as seen in the dihedral angle of 65.92\u2005(12)\u00b0 between the planes through the CN 12H15N3O5S, a tris\u00adubstituted thio\u00adurea derivative, the central CN2S chromophore is almost planar (r.m.s. deviation = 0.018\u2005\u00c5) and the pendant hy\u00addroxy\u00adethyl groups lie to either side of this plane. While to a first approximation the thione-S and carbonyl-O atoms lie to the same side of the mol\u00adecule, the S\u2014C\u2014N\u2014C torsion angle of \u221247.8\u2005(2)\u00b0 indicates a considerable twist. As one of the hy\u00addroxy\u00adethyl groups is orientated towards the thio\u00adamide residue, an intra\u00admolecular N\u2014H\u22efO hydrogen bond is formed which leads to an S(7) loop. A further twist in the mol\u00adecule is indicated by the dihedral angle of 65.87\u2005(7)\u00b0 between the planes through the CN2S chromophore and the 4-nitro\u00adbenzene ring. There is a close match between the experimental and gas-phase, geometry-optimized (DFT) mol\u00adecular structures. In the crystal, O\u2014H\u22efO and O\u2014H\u22efS hydrogen bonds give rise to supra\u00admolecular layers propagating in the ab plane. The connections between layers to consolidate the three-dimensional architecture are of the type C\u2014H\u22efO, C\u2014H\u22efS and nitro-O\u22ef\u03c0. The nature of the supra\u00admolecular association has been further analysed by a study of the calculated Hirshfeld surfaces, non-covalent inter\u00adaction plots and computational chemistry, all of which point to the significant influence and energy of stabilization provided by the conventional hydrogen bonds.In the title compound, C Such N,N\u2032-di-N\u2032-benz\u00adoyl\u00adthio\u00adurea derivatives have a carbonyl group connected to the thio\u00adurea framework and offer opportunities for rich coordination chemistry as these mol\u00adecules feature both hard (oxygen) and soft (sulfur) donor atoms along with nitro\u00adgen donors and indeed, a variety of coordination modes have been observed. The neutral mol\u00adecule has been observed to coordinate in a monodentate-S mode inter\u00adactions \u00b0 is consistent with a significant twist in the mol\u00adecule about the C1\u2014N2 bond; the O3\u2014C6\u2014N2\u2014C1 torsion angle is \u22123.6\u2005(2)\u00b0.Selected geometrical data for (I)2S plane (r.m.s. deviation = 0.017\u2005\u00c5). Crucially, the O1-hydroxy\u00adethyl group is folded towards the thio\u00adamide residue, which allows for the formation of an intra\u00admolecular N2\u2014H\u22efO1 hydrogen bond and an S(7) loop, Table\u00a022S atoms and the terminal C7\u2013C12 aryl ring. From Table\u00a01The hy\u00addroxy\u00adethyl groups lie to either side of the CNd,p) basis set \u00b0 (X-ray) versus \u221216.5\u00b0 , indicating a greater deviation from the anti-disposition in the optimized structure. Also, the N1\u2014C2\u2014C3\u2014O1 and N1\u2014C4\u2014C5\u2014O2 torsion angles are close to symmetric in the optimized structure cf. the experimental structure. Similar trends were noted in analogous calculations performed on the 4-methyl analogue . The layers are connected into a three-dimensional architecture by methyl\u00adene-C\u2014H\u22efO(carbon\u00adyl), methyl\u00adene-C\u2014H\u22efS(thione) and comparatively rare nitro-O\u22ef\u03c0(ar\u00adyl) contacts, Fig.\u00a03b).In the crystal of (I)s Table\u00a02 lead to Crystal Explorer 17 reflects the relative likelihood of the formation of X-to-Y inter\u00adactions in a crystal, i.e. the ratio between the proportion of actual contacts in a crystal to the theoretical proportion of random contacts. Data for (I)The enrichment ratio (ER) descriptor, which is derived from the analysis of the Hirshfeld surface (Jelsch a)\u2013(f), respectively, with a summary of the percentage contributions from the various contacts given in Table\u00a05de + di \u223c1.8\u2005\u00c5 in Fig.\u00a07c). The prominent features in Fig.\u00a07d) reflect the significant C\u22efH/H\u22efC contacts evident in the packing, Tables 2de + di \u223c2.1\u2005\u00c5 in the fingerprint plot of Fig.\u00a07e). The 5.8% contribution from C\u22efO/O\u22efC contacts and the aforementioned ER value of 1.66 clearly indicate the significance of the nitro-N\u2014O\u22ef\u03c0 inter\u00adaction upon the packing; this inter\u00adaction is reflected in the pair of short spikes de + di \u223c3.0\u2005\u00c5, Fig.\u00a07f).The overall fingerprint plots for (I)EBSSEint) are listed in Table\u00a06et al., 2019The energy calculations were performed using DFT-wB97XD/aug-cc-pVTZ of all pairwise inter\u00adactions sums to \u221245.89 kcal/mol, while the total dispersion energy term (Edispersion) computes to \u221251.51 kcal/mol.From the aforementioned, the mol\u00adecular packing is clearly governed by directional hydrogen bonding between mol\u00adecules. The simulated energy frameworks N(H)C(=S)N(CH2CH2OH)2, namely Y = H, which has been reported twice , 68.96\u2005(12), 69.51\u2005(8) and 72.15\u2005(10)\u00b0 for (I)Y = H, F and Me, respectively.There are three literature precedents to (I)The mol\u00adecular packing in the crystals is also very similar with the formation of the intra\u00admolecular thio\u00adamide-N\u2014H\u22efO(hy\u00addroxy) hydrogen bond as well as the inter\u00admolecular hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) hydrogen and hy\u00addroxy-O\u2014H\u22efS(thione) hydrogen bonding, leading to a supra\u00admolecular layer in each case.\u22121): 3277 , 3170 , 3077 , 2973\u20132882 , 1692 , 1538 , 1524 , 1343 , 1270 , 1053 , 734 . UV : \u03bbmax nm (log \u220a) 366.4 (4.16), 301.6 (4.88), 271.2 (5.00), 205.8 (5.14).Synthesis of (I)2, equivalent a 15% weight loss, in the first stage in the range 194 and 222\u00b0C. This was followed by the liberation of a benzene mol\u00adecule, corresponding to 29% weight loss, between 222 and 282\u00b0C, whereas the subsequent stages involve the pyrolysis of CO (282 to 360\u00b0C) and OH (360 to 496\u00b0C) corresponding to 15 and 11% weight loss, respectively. Gradual weight loss continued beyond 800\u00b0C.The pyrolytic process for (I)Uiso(H) set to 1.2Ueq(C). The O- and N-bound H atoms were located from a difference map and refined with O\u2014H and N\u2014H = 0.84\u00b10.01 and 0.88\u00b10.01\u2005\u00c5, respectively, and with Uiso(H) = 1.5Ueq(O) and 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S2056989019017328/hb7880sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019017328/hb7880Isup2.hklStructure factors: contains datablock(s) I. DOI: 1919879CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title morpholino\u00adchlorin adopts a ruffled conformation of its porphyrinic \u03c0-system chromophore inducing a red-shift of its optical spectrum compared to its chlorin analog. 46H16F20N4O3, was crystallized from hexa\u00adne/methyl\u00adene chloride as its 0.44 methyl\u00adene chloride solvate, C46H16F20N4O3\u00b70.44CH2Cl2. The morpholino\u00adchlorin was synthesized by stepwise oxygen insertion into a porphyrin using a \u2018breaking and mending strategy\u2019: NaIO4-induced diol cleavage of the corresponding 2,3-di\u00adhydroxy\u00adchlorin with in situ methanol-induced, acid-catalyzed intra\u00admolecular ring closure of the inter\u00admediate secochlorins bis\u00adaldehyde. Formally, one of the pyrrolic building blocks was thus replaced by a 2,3-di\u00admeth\u00adoxy\u00admorpholine moiety. Like other morpholino\u00adchlorins, the macrocycle of the title compound adopts a ruffled conformation, and the modulation of the porphyrinic \u03c0-system chromophore induces a red-shift of its optical spectrum compared to its corresponding chlorin analog. Packing in the crystal is governed by inter\u00adactions involving the fluorine atoms of the penta\u00adfluoro\u00adphenyl substituents, dominated by C\u2014H\u22efF inter\u00adactions, and augmented by short fluorine\u22effluorine contacts, C\u2014F\u22ef\u03c0 inter\u00adactions, and one severely slipped \u03c0-stacking inter\u00adaction between two penta\u00adfluoro\u00adphenyl rings. The solvate methyl\u00adene chloride mol\u00adecule is disordered over two independent positions around an inversion center with occupancies of two \u00d7 0.241\u2005(5) and two \u00d7 0.199\u2005(4), for a total site occupancy of 88%.The title morpholino\u00adchlorin, C Those adjacent to the morpholine moiety (C6F5 rings of C21 and C39) are more sterically encumbered and are about 10\u00b0 closer to perpendicular to the macrocycle plane, with values of 82.70\u2005(3) and 81.44\u2005(2)\u00b0. The corresponding values for compound 1b are very similar, with values of 71.89 and 73.73\u00b0, and 89.55 and 86.32\u00b0, respectively.Similar to other 1b and 1d allows us to investigate how minor conformational changes might affect the optical properties of the morpholino\u00adchlorins. The torsion angles between the two C\u2014C bonds in the morpholine units in the two morpholino\u00adchlorins 1b and 1d vary slightly, with this angle being smaller in the title compound [35.2\u00b0 in 1b and 25.5\u2005(4)\u00b0 1d]. This angle is important as it strongly affects the \u03bbmax of the morpholino\u00adchlorins .The close structural relationship between the 2,3-disubstituted derivatives 1d in the crystal. Dominant are C\u2014H\u22efF hydrogen-bond-like inter\u00adactions, involving both methyl as well as pyrrole moieties as the hydrogen-atom donor. Weak C\u2014H\u22efF inter\u00adactions involving the solvent are also present. The most prominent of these inter\u00adactions are given in Table\u00a01Inter\u00adactions involving fluorine atoms play a dominant role in facilitating the arrangement of mol\u00adecules of et al., 2019a). Both meth\u00adoxy substituents are engaged in several of these inter\u00adactions: C45 exhibits inter\u00adactions with fluorine atoms from three different penta\u00adfluoro\u00adphenyl groups: with meta fluorine atoms F12iii and F19iv , and one intra\u00admolecular inter\u00adaction with F20, an ortho-fluorine atom. Angular and H\u22efF distance values for this intra\u00admolecular inter\u00adaction appear quite unfavorable: the C\u2014H\u22efF angle is only 103\u00b0, and the H\u22efF distance is 2.82\u2005\u00c5. However, only a slight rotation of the methyl H atoms is required to create a much more favorable geometry, and the C\u22efF distance between C45 and F20 is at 3.184\u2005(3) quite short . Inter\u00adactions involving the meth\u00adoxy group of C46 involve F10v and F1vi, two ortho-fluorine atoms . Two C\u2014H\u22efF inter\u00adactions originate from pyrrole moieties, involving H atoms at the pyrrole moieties flanking the morpholine unit: H8 towards F18i, and H18 towards F8ii, with both F8 and F18 being para-fluorine atoms . These two inter\u00adactions work in tandem with each other and with a severely slipped \u03c0\u2013\u03c0 stacking inter\u00adaction, between the rings of F6\u2013F10 and F16i\u2013F20i, connecting two opposite ends of the morpholino\u00adchlorin mol\u00adecule with its neighbors to create infinite chains connected via C\u2014H\u22efF and slipped \u03c0\u2013\u03c0 stacking inter\u00adactions . The centroid-to-centroid distance of the \u03c0-stacking inter\u00adaction is 4.3551\u2005(15)\u2005\u00c5, with a ring slippage of 2.795\u2005\u00c5 and a centroid-to-mean-plane distance of 3.1661\u2005(12)\u2005\u00c5. The last C\u2014H\u22efF inter\u00adaction involves the methyl\u00adene group of the minor moiety solvate methyl\u00adene chloride mol\u00adecule. Given the degree of disorder of the solvate mol\u00adecules (see Refinement section), this inter\u00adaction is probably vaguely defined at best and will not be discussed in detail.The most prominent C\u2014H\u22efF inter\u00adactions , inter\u00adactions between two fluorine atoms are different and much weaker in nature \u2005\u00c5 (F5\u22efF7vii) and 2.828\u2005(3)\u2005\u00c5 (F5\u22efF8vii) . With the intra\u00admolecular distance between F7 and F8 being 2.726\u2005(2)\u2005\u00c5, this leads to the formation of a nearly equilateral triangle of F atoms . It should be noted that atom F8 of this F3-triangle also acts as the acceptor of the C18\u2014H18\u22efF8ii contact and the backside of the aromatic ring of F8 is involved in the slipped \u03c0\u2013\u03c0 stacking inter\u00adaction (see discussion above). Fluorine atom F11 features a close contact with a symmetry-created copy of itself, created by a twofold axis. The F\u22efFiii distance here is 2.783\u2005(3)\u2005\u00c5, and the C\u2014F\u22efFiii angle is 125.5\u2005(2)\u00b0 .Besides C\u2014H\u22efF inter\u00adactions, which are generally considered as directional inter\u00adactions similar in strength to the better investigated C\u2014H\u22efO inter\u00adactions, ms Fig.\u00a06a. It shns Fig.\u00a07a. The ler Fig.\u00a06b. The Fvi and C42vi) of another penta\u00adfluoro\u00adphenyl ring [symmetry code: (vi) \u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a01]. The F\u22efC distances are 3.034\u2005(3) and 2.978\u2005(3)\u2005\u00c5 for F2 and F3, respectively. There are two inter\u00adactions of this kind per mol\u00adecule, one as the C\u2014F donor and one as the \u03c0-density moiety accepting the C\u2014F\u22ef\u03c0 bond, connecting mol\u00adecules into centrosymmetric dimers. One of the methyl C\u2014H\u22efF contacts (towards F1) is also involved in the formation of these dimers .Besides F11, F2 and F3 are also involved in inter\u00admolecular C\u2014F\u22ef\u03c0 inter\u00adactions, pointing nearly perpendicularly towards C atoms , fifteen morpholino\u00adchlorins, two thio\u00admorpholines . Disorder with hexane, the other type of solvent used during crystallization, was excluded as a possibility due to the limited size of the solvate pocket, and it is thus assumed that 12% of void spaces in the crystal structure remained unoccupied during the crystallization process.The solvate methyl\u00adene chloride mol\u00adecule is disordered over four positions around an inversion center . The C\u2014Cl and Cl\u22efCl distances were restrained to target values and 2 and CH3 moieties, respectively. Methyl CH3 groups were allowed to rotate but not to tip to best fit the experimental electron density. Uiso(H) values were set to a multiple of Ueq(C) with 1.5 for CH3, and 1.2 for CH and CH2 units, respectively.N-bound H atoms were located in a difference electron-density map and were freely refined. H atoms attached to carbon atoms were positioned geometrically and constrained to ride on their parent atoms. C\u2014H bond distances were constrained to 0.95\u2005\u00c5 for pyrrole CH moieties, and to 1.00, 0.99 and 0.98\u2005\u00c5 for aliphatic CH, CH10.1107/S2056989020009093/is5543sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989020009093/is5543Isup2.hklStructure factors: contains datablock(s) I. DOI: 2013871CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Phormidium ambiguum and Microcystis aeruginosa) were exposed to diurnal light-intensity variation to investigate their favorable and stressed phases during a single day. The photosynthetically active radiation (PAR) started at 0 \u00b5mol\u00b7m\u22122\u00b7s\u22121 (06:00 h), increased by ~25 \u00b5mol\u00b7m\u22122\u00b7s\u22121 or ~50 \u00b5mol\u00b7m\u22122\u00b7s\u22121 every 30 min, peaking at 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 or 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 (12:00 h), and then decreased to 0 \u00b5mol\u00b7m\u22122\u00b7s\u22121 (by 18:00 h). The H2O2 and antioxidant activities were paralleled to light intensity. Higher H2O2 and antioxidant levels , and superoxidase dismutase) were observed at 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 rather than at 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121. Changes in antioxidant levels under each light condition differed between the species. Significant correlations were observed between antioxidant activities and H2O2 contents for both species, except for the CAT activity of P. ambiguum at 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121. Under each of the conditions, both species responded proportionately to oxidative stress. Even under maximum light intensities (300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 or 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 PAR intensity), neither species was stressed. Studies using extended exposure durations are warranted to better understand the growth performance and long-term physiological responses of both species.Two harmful cyanobacteria species ( The growth and spread of cyanobacteria have increased, thus threatening today\u2032s water bodies and supplies worldwide ,2. GlobaDuring cyanobacteria control efforts, chemical control measures are discouraged due to their potentially harmful secondary effects on ecosystems ,13,14, wBiological, chemical, and physical factors collectively determine the occurrence and distribution of cyanobacteria in the environment ,25,26. PThe photosynthetic species produce reactive oxygen species (ROS) as a byproduct of the photosynthesis process, which is harmful when accumulated in cells . Therefo2O2) and antioxidant (guaiacol peroxidase (GPX), catalase (CAT), ascorbic peroxidase (APX), and superoxidase dismutase (SOD)) responses of cyanobacteria to diurnal changes in the light intensity were studied. Two photosynthetically active radiation (PAR) levels, 300 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121, were selected as maximum light intensities, and the responses of two cyanobacterial species, Phormidium ambiguum and Microcystis aeruginosa, were tested for gradually varying light conditions. P. ambiguum is a non-heterocystous filamentous cyanobacterial species, categorized as a benthic cyanobacterium [M. aeruginosa is a floating (buoyant) and colony foaming type cyanobacterium [In this study, the oxidative stress and M. aeruginosa (strain NIES 111) were obtained from the National Institute for Environmental Studies, Tsukuba, Japan. Both species were cultured for 14 days at 20 \u00b0C under a 12 h:12 h light:dark cycle inside an incubator . Light was provided with cool white fluorescent lamps and the intensity was maintained at 20\u201330 \u00b5mol\u00b7m\u22122\u00b7s\u22121 PAR. The nutrient medium was 100% BG-11 [0% BG-11 . During P. ambiguum and M. aeruginosa cyanobacteria cultures were made, maintaining the 0.6 \u00b1 0.02 optical density measured at 730 nm (OD730) using a spectrophotometer . The dilution of the cyanobacteria culture was accomplished with BG11 nutrient medium. In all experiments, the temperature was maintained at 20 \u00b0C in an incubator, whereas the lighting conditions changed from 0 \u00b5mol\u00b7m\u22122\u00b7s\u22121 (at 06:00 h) to 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 or 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 (at 12:00 h) by changing the lighting intensity by ~25 \u00b5mol\u00b7m\u22122\u00b7s\u22121 or ~50 \u00b5mol\u00b7m\u22122\u00b7s\u22121 every 30 min with a VBP-L24-C2 light . The light intensity was then decreased at the same rate (until 18:00 h). The lighting condition was controlled with warm light-emitting diode panel lights, and the light intensity was measured using a quantum flux meter . Cyanobacteria samples from each flask were collected for analysis every 3 h, at 06:00 h, 09:00 h, 12:00 h, 15:00 h, 18:00 h, and 21:00 h. To facilitate mixing, each flask was manually shaken at the time of sampling.Following the 14-day incubation, 3 replicate conical flasks from each of the 2O2 contents were estimated according to standard methods [g for 10 min at 4 \u00b0C. The cell pellets were washed once with ultrapure water . To extract cellular H2O2, cell pellets were homogenized in 1 mL of 0.1 M pH 6.5 phosphate buffer and centrifuged at 10,000\u00d7 g for 10 min at 4 \u00b0C. A total of 750 \u03bcL of 1% titanium chloride in 20% H2SO4 (v/v) was then added to initiate the reaction. The optical absorption was measured at 410 nm using a spectrophotometer (UVmini-1240), following centrifugation at room temperature (25 \u00b1 2 \u00b0C). The H2O2 concentration was determined using a standard curve, prepared using a series of samples with known H2O2 concentration.Cellular H methods . Brieflyg at 4 \u00b0C for 10 min and removing the supernatant and cell pellets, which were homogenized in 1 mL potassium phosphate buffer . A total of 65 \u00b5l of enzyme extract was then mixed with 920 \u03bcL of potassium phosphate buffer containing 20 mM guaiacol. With the addition of 15 \u03bcL of 0.6% H2O2, the reaction was then started, and the absorbance change was recorded at 470 nm every 10 s for 3 min using UV mini-1240. GPX activity was calculated using an extinction coefficient of 26.6 mM/cm.The GPX activity was assayed as described by Hoda et al. and MacAg at 4 \u00b0C for 10 min. The supernatant was removed, and the cell pellets were homogenized in 1 mL potassium phosphate buffer , containing 0.1 mM EDTA. After centrifuging again , the supernatant was collected as the enzyme extract. The CAT activity was measured by reacting 15 \u00b5L of 750 mM H2O2, 920 \u00b5L of potassium phosphate buffer, and 65 \u00b5L of extract supernatant. Optical absorption was measured at 240 nm using UV mini-1240. The measurements were recorded every 10 s for 3 min, and the CAT activity was calculated using an extinction coefficient of 39.4 mM/cm.CAT activity was measured using the method described by Aebi . A total2O2. Calculations were performed using a molar extinction coefficient for ascorbate of 2.8 mM/cm.APX activity was assayed, as described by Nakano and Asada . The decSOD activities were determined by performing nitro blue tetrazolium (NBT) assays, as described by Ewing and Janero . Each saP. ambiguum and M. aeruginosa were evaluated using a Student\u2019s t-test, assuming equality of variance. Pearson\u2019s correlation analysis was used to evaluate correlations between parameters. Statistical analyses were performed by using IBM SPSS Statistics for Windows, Version 25.0. .One-way analysis of variance (ANOVA), followed by Tukey\u2019s post-hoc test, was performed to test the statistical significance of variations among the means of sample groups. Data were normalized relative to the starting group (06:00 h), by dividing the results of each group by the corresponding 06:00 h group for each replicate. Significant differences between experimental groups of 2O2 contents of P. ambiguum and M. aeruginosa increased with increasing light intensity and peaked between 12:00 h and 15:00 h. The H2O2 content decreased thereafter in parallel with decreasing light intensity. The cyanobacteria were exposed to dark at 18:00 h; however, even at 21:00 h, the H2O2 contents did not reach the initial level measured at 06:00 h . ANOVA testing of P. ambiguum exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped H2O2 contents at 06:00 h, 09:00 h, and 18:00 h, and 12:00 h, 15:00 h, and 21:00 h . For M. aeruginosa exposed to 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity, ANOVA testing grouped H2O2 contents at 06:00 h, 09:00 h, and 21:00 h, 12:00 h and 15:00 h, and 18:00 h . ANOVA testing of M. aeruginosa exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped H2O2 contents at each time point . Comparing the H2O2 contents of 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121, max PAR intensity groups showed that the H2O2 contents of both P. ambiguum and M. aeruginosa differed significantly at each time point (p < 0.01 for each time point).The H 06:00 h . For P. P. ambiguum and M. aeruginosa increased over time and reached the maximum at 12:00 h when the maximum PAR intensity was reached (300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 or 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121). With decreasing light intensity, the GPX activities of both species were decreased. However, with the 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity group, at 18:00 h and 21:00 h, the GPX activities decreased even further than the starting GPX activity (06:00 h). With the 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR-intensity, the GPX activity of both species decreased with decreasing light, but the GPX activity of P. ambiguum remained higher than the starting GPX activity, even at 21:00 h. For M. aeruginosa, GPX activity of 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity reached the starting GPX activity (06:00 h) at 21:00 h . ANOVA testing of the P. ambiguum exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped GPX activities at 06:00 h and 09:00 h; 12:00 h and 15:00 h; 18:00 h; and 21:00 h . For M. aeruginosa exposed to 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity, ANOVA testing groped GPX activities at 06:00 h, 18:00 h, and 21:00 h; 12:00 h; and 09:00 h and 15:00 h . ANOVA testing of M. aeruginosa exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped GPX activities at 06:00 h and 21:00 h; 06:00 h, 09:00 h, and 18:00 h; 09:00 h, 15:00 h, and 18:00 h; and 12:00 h . A comparison between 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity groups showed that the GPX activities of P. ambiguum and M. aeruginosa differed significantly at each time point (p < 0.01 for each time point).The GPX activities of both 21:00 h . For P. P. ambiguum and M. aeruginosa increased over time but showed a delayed response, as the maximum CAT activities were reached at 15:00 h (which was 3 h after the maximum light intensities of 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 or 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 PAR was reached). Decreasing light intensities were paralleled by reduced CAT activities, although the CAT activity did not reach the initial CAT level, even at 21:00 h . ANOVA testing of P. ambiguum exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped CAT activities at 06:00 h and 21:00 h; 09:00 h, 12:00 h, and 18:00 h; and 15:00 h . For M. aeruginosa exposed to 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity, ANOVA testing groped CAT activities at 06:00 h and 21:00 h; 9:00 h; 21:00 h, 09:00 h, and 18:00 h; and 12:00 h and 15:00 h . ANOVA testing of M. aeruginosa exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped CAT activities at 06:00 h and 21:00 h; 09:00 h and 18:00 h; 12:00 h; and 15:00 h . Comparisons between the 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity groups showed that the CAT activities of P. ambiguum differed significantly at 09:00 h, 12:00 h, 15:00 h, and 18:00 h (p < 0.01), although the CAT activities at 06:00 h and 21:00 h were not different (p > 0.05). The CAT activities of 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity groups of M. aeruginosa also differed at 09:00 h, 15:00 h, and 21:00 h (p < 0.05).The CAT activities of both 21:00 h . For P. P. ambiguum and M. aeruginosa increased with increasing light intensity, but a delayed response was observed, where the maximum APX activity was reached at 15:00 h (3 h after the maximum light intensities were reached). With subsequent decreasing light intensity, the APX activities of both species decreased and reached the initial APX activity (06:00 h) at 21:00 h . ANOVA testing of P. ambiguum exposed 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped APX activities at 06:00 h, 18:00 h, and 21:00 h; 09:00 h, 18:00 h, and 21:00 h; and 12:00 h and 15:00 h . For M. aeruginosa exposed to 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity, ANOVA testing grouped APX activities at 06:00 h, 18:00 h, and 21:00 h; 06:00 h, 09:00 h, and 15:00 h; and 12:00 h . ANOVA testing of M. aeruginosa exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped APX activities at 06:00 h and 21:00 h; 06:00 h, 09:00 h, and 18:00 h; 09:00 h, 15:00 h, and 18:00 h; and 12:00 h . Comparisons between the 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity groups showed that the differences in APX activities for both species were significantly higher in the 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity groups, from 09:00 h to 18:00 h (p < 0.01 for each light condition for both species). The 21:00 APX activities of P. ambiguum were significantly lower in the 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity group than in the 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity group (p < 0.01), although no differences were observed for M. aeruginosa.The APX activities of both 21:00 h . For P. P. ambiguum and M. aeruginosa increased with the light intensity. With decreasing light, SOD activity decreased for both species and approached the starting level at 21:00 h, for both 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR conditions . ANOVA testing of P. ambiguum exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped SOD activities at 06:00 h, 18:00 h, and 21:00 h; 09:00 h, 18:00 h, and 21:00 h; 12:00 h; and 15:00 h . For M. aeruginosa exposed to 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity, ANOVA testing grouped SOD activities at 06:00 h and 21:00 h; 06:00 h and 09:00 h; 09:00 h, 15:00 h, and 18:00 h; and 12:00 h, 15:00 h, and 18:00 h . ANOVA testing of M. aeruginosa exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped SOD activities at 06:00 h and 21:00 h; 06:00 h, 09:00 h, 15:00 h, and 18:00 h; and 09:00 h and 12:00 h . Comparisons between the 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity groups indicated that the SOD activities of P. ambiguum differed at 12:00 h and 15:00 h (p < 0.01) and that M. aeruginosa showed no significant differences between 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity conditions (p > 0.05).The SOD activities of both nditions . For P. P. ambiguum and M. aeruginosa increased with the light intensity. With decreasing light, the AOX activity decreased, and, for both species, the AOX activity approached the starting level at 21:00 h, under both the 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 PAR conditions . ANOVA testing of P. ambiguum exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped AOX activities at 06:00 h, 18:00 h, and 21:00 h; 09:00 h, 18:00 h, and 21:00 h; 15:00 h; and 12:00 h . For M. aeruginosa exposed to 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity, ANOVA testing grouped AOX activities at 06:00 h and 21:00 h; 09:00 h and 18:00 h; and 12:00 h and 15:00 h . ANOVA testing of M. aeruginosa exposed to 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity grouped AOX activities at 06:00 h and 21:00 h; 06:00 h and 18:00 h; 09:00 h, 15:00 h, and 18:00 h; and 12:00 h and 15:00 h . The AOX activities of 300 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensities differed significantly at 12:00 h and 15:00 h, with P. ambiguum (p < 0.01), while those for M. aeruginosa only exhibited a significant difference at 09:00 h (p < 0.01). In other cases, no significant differences were observed.The total antioxidant (AOX) activities of both nditions . For P. 2O2 levels and those of the antioxidants and the AOX levels were significantly linearly correlated, except for the GPX activity of P. ambiguum under the 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 max PAR intensity , confirming that less variance occurred. When the AOX values were considered, the 2R showed relatively higher values (>0.72).The relationships between the Hntensity . Pearson2O2 contents and the antioxidant activities of P. ambiguum and M. aeruginosa were highly responsive to the diurnal variations in light intensity. In this study, the only variable factor was the light intensity, where H2O2 levels were high during times of higher light intensities and decreased at lower light intensities. When cellular H2O2 level increases, the antioxidant activities correspondingly increase to prevent damage induced by oxidative stress [2O2 levels, the antioxidant activities also varied during the same time frame and followed the H2O2 levels, which increased at higher light intensities and decreased at lower light intensities. The antioxidant activities of both species were correlated linearly with the H2O2 contents. Although the H2O2-antioxidant relationships were varied from strong to weak (depending on the antioxidant species), overall, our findings suggest that the antioxidant levels of both species responded to the cellular H2O2 level accordingly.The He stress ,50. As o2O2 and antioxidant responses followed the same trends for both maximum light intensity conditions (PAR intensities of 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121 or 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121). Under the maximum PAR intensity of 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121, the cyanobacteria received approximately twice the photon energy of the group with a maximum PAR of 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121. Therefore, it can be anticipated that the experiment groups, which receive higher photon energy, undergo an enhanced rate of photosynthesis. This is evidenced by the increased H2O2 formed after exposure to a higher light intensity [2O2 production is reduced with higher light exposure [2O2 contents correlated directly with light intensity, even at higher intensities, PAR intensities under 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121 did not subject either cyanobacterial species to photo stress. However, this study only involved a single day diurnal variation, and the H2O2 levels of the cells did not reach the starting H2O2 conditions at 06:00 h (even at 21:00 h) for either species. The antioxidant activities almost decreased to the initial conditions by 21:00 h. Therefore, cells may undergo oxidative stress during dark conditions due to the lack of antioxidant activities. If the H2O2 was continued to be presence in cells, the protein synthesis of photosystems will be inhibited [2O2 and adaptation responses.The Hntensity ,52. Howentensity ,54, duriexposure ,56,57. Anhibited and, in nhibited . TherefoM. aeruginosa than P. ambiguum, except for GPX. Under high H2O2 contents, the AOX activity was highly elevated in P. ambiguum, but in the dark, both species reached the starting AOX activity level at 21:00 h. This finding suggests that M. aeruginosa is less tolerant to oxidative stress than P. ambiguum [2O2 contents, both species demonstrated significant linear relationships (with the only exception being for GPX of P. ambiguum under a maximum PAR of 300 \u00b5mol\u00b7m\u22122\u00b7s\u22121). Therefore, despite the high AOX content of the P. ambiguum, both species were able to maintain balanced antioxidant activity under every light condition of the single day exposure.The antioxidant levels differed between the two species, where the response level was lower for ambiguum ,59. ConcM. aeruginosa is a buoyant species that floats in a range of depths and might have higher tolerance to oxidative stress [P. ambiguum. However, in the present study, both species experienced same light intensity variance as the cultures were mixed periodically. The non-different H2O2 contents between the two species suggested that both species experienced similar levels of oxidative stress. Therefore, less oxidative stress tolerance of P. ambiguum triggered the antioxidant activity at a relatively higher rate. Conversely, nonenzymatic antioxidants, primarily carotenoids, protect against ROS in phototrophs, including cyanobacteria [P. ambiguum than M. aeruginosa [M. aeruginosa is due to involvement of nonenzymic antioxidant over the P. ambiguum.The difference in antioxidant levels of the two species can be related to their behavioral characteristics. The e stress ,61 than bacteria ,62. The ruginosa ,64,65; t\u22122\u00b7s\u22121 and 600 \u00b5mol\u00b7m\u22122\u00b7s\u22121) confirmed that the OD730 and chlorophyll-a contents of cyanobacteria were significantly reduced, which was associated with oxidative stress [Our previous study on the effects of 8 days of exposure to non-varying, high-light intensities (300 \u00b5mol\u00b7me stress . In futu"} +{"text": "V\u03b39V\u03b42 T\u00a0cells respond in a TCR-dependent fashion to both microbial and host-derived pyrophosphate compounds . Butyrophilin-3A1 (BTN3A1), a protein structurally related to the B7 family of costimulatory molecules, is necessary but insufficient for this process. We performed radiation hybrid screens to uncover direct TCR ligands and cofactors that\u00a0potentiate BTN3A1\u2019s P-Ag sensing function. These experiments identified butyrophilin-2A1 (BTN2A1) as essential to V\u03b39V\u03b42 T cell recognition. BTN2A1 synergised with BTN3A1 in sensitizing P-Ag-exposed cells for V\u03b39V\u03b42 TCR-mediated responses. Surface plasmon resonance experiments established V\u03b39V\u03b42 TCRs used germline-encoded V\u03b39 regions to directly bind the BTN2A1 CFG-IgV domain surface.\u00a0Notably, somatically recombined CDR3 loops implicated in P-Ag recognition were uninvolved. Immunoprecipitations demonstrated close cell-surface BTN2A1-BTN3A1 association independent of P-Ag stimulation. Thus, BTN2A1 is a BTN3A1-linked co-factor critical to V\u03b39V\u03b42 TCR recognition. Furthermore, these results suggest a composite-ligand model of P-Ag sensing wherein the V\u03b39V\u03b42 TCR directly interacts with both BTN2A1 and an additional ligand recognized in a CDR3-dependent manner. \u2022Radiation hybrids identify BTN2A1 as crucial for V\u03b39V\u03b42 phosphoantigen (P-Ag) sensing\u2022BTN2A1 binds directly to the T\u00a0cell receptor via germline-encoded regions of V\u03b39\u2022Cell-surface BTN2A1 associates directly with BTN3A1 independent of P-Ag stimulation\u2022The V\u03b39-BTN2A1 interaction modality suggests an additional CDR3-dependent TCR ligand Karunakaran et\u00a0al. find that butyrophilin 2A1 (BTN2A1) associates with BTN3A1 on the cell surface and binds directly to germline-encoded regions of the V\u03b39 chain of the V\u03b39V\u03b42 TCR. Thus, BTN2A1 collaborates with BTN3A1 to potentiate V\u03b39V\u03b42 T\u00a0cell recognition, playing an essential role in phosphoantigen sensing. Human peripheral blood \u03b3\u03b4 T\u00a0cells are dominated from an early age by V\u03b39V\u03b42 lymphocytes , an innaV\u03b39V\u03b42-mediated P-Ag sensing requires cell-cell contact and depeTursiops truncatus) and alpaca (Vicugna pacos) . Of notea pacos) . In conta pacos) . Consista pacos) . Furthera pacos) and canca pacos) . From thOur previous studies have established that BTN3A1 is necessary but not sufficient for P-Ag sensing and indicated the existence of an additional putative Chr-6-encoded factor that synergized with BTN3A1 to stimulate P-Ag-mediated responses , which wWe showed previously that a T\u00a0cell hybridoma expressing the\u00a0V\u03b39V\u03b42 MOP TCR produced interleukin (IL)-2 in co-culture with BTN3A1-transduced Chinese hamster ovary (CHO) cells incubated with the anti-BTN3A1 monoclonal antibody (mAb) 20.1 but exhibited a complete lack of response to HMBPP or Zol . In contTo identify Factor X, we used an unbiased genome-based approach involving generation of radiation hybrids between CHO-Chr 6 cells and BTN3A1-transduced hypoxanthine-aminopterin-thymidine (HAT)-sensitive rodent fusion partners and subsequent analysis of their capacity to stimulate P-Ag sensing by V\u03b39V\u03b42 T\u00a0cells A. We posWe fused CHO-Chr6 cells separately with two HAT-sensitive fusion partners\u2014first BTN3A1-transduced A23 hamster cells and second BTN3A1-transduced mouse BW cells Sanders; resultiHFE, the BTN3A1 gene already transduced into rodent fusion partners, BTN3A2, BTN3A3, and BTN2A1 and BTN2A2. Since we knew that expression of all three BTN3 genes was insufficient for reconstitution of the P-Ag response , BTN2A1 and BTN2A2, which to date have been discussed mainly for their immunomodulatory properties or BTN2A1 (BTN2A1\u2212/\u2212) or BTN2A2 alone (BTN2A2\u2212/\u2212). Inactivation of both BTN2 genes completely abolished interferon (IFN)\u03b3 production by polyclonal V\u03b39V\u03b42 T\u00a0cell lines in response to Zol\u00a0pulsed cells (BTN2\u2212/\u2212 and BTN2A1\u2212/\u2212 exhibited a complete loss of IL-2 production by TCR-MOP cells in response to either HMBPP (BTN2A2\u2212/\u2212 were similar to wild-type (WT) 293T cells and 49.9\u00a0\u03bcM (n\u00a0= 8), respectively . Injection of BTN2A1 IgV produced substantially enhanced signals over surfaces with immobilized V\u03b39V\u03b42 TCR relative to V\u03b34V\u03b45 and V\u03b32V\u03b41 TCRs or control streptavidin surfaces, indicating specific binding A. Equiliectively B.Figure\u00a0+ T\u00a0cell subset (d 46.6\u03bcM [n\u00a0= 8]) of the BTN2A1-V\u03b39V\u03b41 TCR interaction (d 39\u201350\u00a0\u03bcM [n\u00a0= 3]).An initial homology model suggested strong feasibility of a similar interaction mode and highlighted seven amino acids on the face of the BTN2A1 IgV domain incorporating the C, C', F and G \u03b2 strands (CFG face), equivalent to the region of BTNL3 IgV domain involved in binding V\u03b34, as candidates for alanine mutation C. Indivi0\u201315\u00a0\u03bcM) D and S3B0\u201315\u00a0\u03bcM) . Based o0\u201315\u00a0\u03bcM) D and a B0\u201315\u00a0\u03bcM) E in the 0\u201315\u00a0\u03bcM) F and S3G0\u201315\u00a0\u03bcM) G and S3H+ TCR binding and assessed effects on V\u03b39V\u03b42-mediated P-Ag response. Although permissive for cell-surface BTN2A1 expression and CDR2 loops of the V\u03b39 IgV domain in BTN2A1 interaction, regions also critical for BTNL3-V\u03b34 interaction Figure\u00a0F. ConsisCollectively, these findings established that V\u03b39V\u03b42 TCR binds BTN2A1 IgV via a binding mode that closely mimics that of V\u03b34 TCR for BTNL3 and that this binding is essential for P-Ag sensing but occurs alongside parallel and essential V\u03b42 CDR-mediated binding events.BTN and BTNL molecules have been shown to form either homo- or heterodimers . To inveInterestingly, the BTN2A1 IgV-C model also highlighted close proximity of extracellular membrane-proximal cysteine residue (C247) with its equivalent residue in the opposing monomer B, suggesFinally, by combining our HADDOCK-derived model of V\u03b39V\u03b42/BTN2A1 interaction B with ouTo assess whether BTN2A1 and BTN3A1 were associated with each other at the cell surface either before or after P-Ag exposure, a membrane-impermeable amine-reactive cross-linker incorporating a 16\u00c5 spacer was used to cross-link proteins on the surface of CHO transductants co-expressing C-terminally HA-tagged BTN2A1 (BTN2A1-HA) and a N-terminally FLAG-tagged BTN3A1 (FLAG-BTN3A1). Immunoprecipitation (IP) using anti-HA beads or 20.1 mAb and subsequent anti-FLAG western blot (WB) was used to detect cross-linked BTN2A1-BTN3A1 species .Figure\u00a05Following IP of BTN2A1-HA using anti-HA beads and subsequent WB detection of FLAG-BTN3A1 under reducing conditions using an anti-FLAG antibody, two discrete bands were detected that considerably exceeded the size of either BTN2A1 or BTN3A1 monomers A; each w1H-15N-labeled BTN3A1 IgV in E.\u00a0coli and performed 1H-15N heteronuclear single quantum coherence (HSQC) spectroscopy in the absence and presence of an equimolar amount of naturally labeled E.\u00a0coli-expressed BTN2A1 IgV . Reagents generated in this study are available on request from the Lead Contact with a completed Materials Transfer Agreement.Further information and requests for resources and reagents should be directed to and will be fulfilled by the Lead Contact, Benjamin E. Willcox were cultured with RPMI (GIBCO) supplemented with 10% FCS, 1\u00a0mM sodium pyruvate, 2.05\u00a0mM glutamine, 0.1\u00a0mM nonessential amino acids, 50\u00a0\u03bcM \u03b2-mercaptoethanol, penicillin (100\u00a0U/mL) and streptomycin (100\u00a0U/mL). Peripheral blood mononuclear cells isolated from healthy volunteers were also maintained as above with or without rhIL-2 (Novartis Pharma). 293T cells were maintained in DMEM (GIBCO) supplemented with 10% FCS.6 or 107 cells) were irradiated at Faxitron CP160 . The irradiated cells and fusion partner (BW or A23) were mixed at 1:1 or 1:3 ratio (irradiated cell:fusion partner) and centrifuged at 461\u00a0g for 5\u00a0min at RT. The cell pellet was gently tapped and 1\u00a0mL PEG1500 was added slowly over a minute with gentle mixing in a prewarmed water-bath. After addition of PEG, cells were resuspended in 50\u00a0mL warm serum free RPMI and incubated for 30\u00a0min, followed by centrifugation at 461\u00a0g for 5\u00a0min and careful resuspension in RPMI supplemented with 10% FCS at 104 cells/mL. The cell suspension was seeded in 96 well plate flat bottom (A23 fused) or round bottom (BW fused) plates in 100\u00a0\u03bcl per well. On the following day, 100\u00a0\u03bcl of 2X HAT was added and cells were selected for two weeks. The selected clones were supplemented with HT medium and further seeded at limiting dilutions to obtain single cell clones which were tested for P-Ag mediated activation of our V\u03b39V\u03b42 TCR (MOP) transductants. P-Ag presentation capable and incapable clones were PCR characterized for human Chr 6 regions with primers listed in CHO Chr 6 using STAR and the corresponding Gene Transfer Format (gtf) file (version 87). Unmapped reads and those exhibiting more than two mismatches were selected and mapped against the Chinese hamster genome . The corresponding gtf-file was downloaded from the pre-Ensembl ftp site (Cricetulus_griseus.CriGri_1.pre.gtf). Afterward, all unmapped reads and those containing more than two mismatches were again selected to finally map against the human genome . Only reads showing maximally one mismatch were considered as true. With the help of featureCounts, mapped reads were assigned to genomic features using the above mentioned gtf-files. The results were summarized within an Excel-file. Further gene information were extracted from BioMart and total RNA was extracted. Sequencing libraries were produced with an Illumina Truseq RNA preparation kit as described by the supplier\u2019s protocol and were sequenced with an Illumina HiSeq4000. Sequence reads were mapped to the human genome (hg38) with STAR (version STAR_2.50a) and read counts of gene transcripts were determined using gtf file Homo_sapiens.GRCH38.84.gtf and featureCount (v1.5.0-p1). Cell lines were then compared for presence, i.e., expression, of human Chr 6 genes. To filter out reads descending from mouse or hamster cells, all fastq-files were initially mapped against the mouse genome as per manufacturer\u2019s protocol.For 6 cells/mL density in a 96 well U bottom plate for 10\u00a0days with 100\u00a0\u03bcL per well. After 10\u00a0days, cells were pooled and washed twice and cultivated to rest without rhIL- 2 at 106 cells/mL density in a 6-well plate. After three days, rested cells were subjected for further experiments.Fresh peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteers after obtained written informed consent in accordance with the Declaration of Helsinki and approval by the University of W\u00fcrzburg institutional review board. Whole blood was layered over the Histopaque-1077 in a 50\u00a0mL falcon tube and centrifuged at 400\u00a0g for 30\u00a0min at room temperature (RT) with no acceleration and brakes. After centrifugation, the opaque interface containing PBMCs were aspirated and washed twice at 461\u00a0g for 5\u00a0min at RT. V\u03b39V\u03b42 T\u00a0cells were expanded by cultivation of PBMCs with RPMI containing 10% FACS, 1\u00a0\u03bcM BrHPP and recombinant human IL-2 100 IU/mL (Novartis Pharma) in 10BTN2A1 and BTN2A2 genes were disrupted in 293T cells using CRISPR. The CRISPR sequencing targeting functional BTN2A genes were designed with the help of online tools mentioned in the table (software section) and sequences were cloned into GeneArt CRISPR Nuclease vector as per manufacturer\u2019s instructions. On day1, 1.5\u00a0\u00d7 106 293T cells were seeded in a 6\u00a0cm cell culture plate with DMEM medium without pyruvate (10% FCS). On day 2, cells were transfected with 5\u00a0\u03bcg of BTN2A-IgV_CRISPR cloned GeneArt CRISPR Nuclease (CD4 enrichment) Vector or BTN2A1_49FCRISPR cloned GeneArt CRISPR Nuclease (OFP Reporter) Vector or BTN2A2_343CRISPR cloned GeneArt CRISPR Nuclease (CD4 enrichment) Vector in a calcium-phosphate dependent method BTN2A1 IgV CRISPR target site2A1IgV \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0GCAGTGTTTGTGTATAAAGGTGGCAGAGAGAGAACAGAGGAGCAGATGGAGGAGT \u00a0552A1allele1 \u00a0GCAGTGTTT\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014-GTGGCAGAGAGAGAACAGAGGAGCAGATGGAGGAGT \u00a0452A1allele2 \u00a0GCA\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014-GTGGCAGAGAGAGAACAGAGGAGCAGATGGAGGAGT \u00a039\u2217\u2217\u2217 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0BTN2A2 IgV CRISPR target site1b) 2A2IgV \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0GCAGTGTTTGTGTATAAGGGTGGGAGAGAGAGAACAGAGGAGCAGATGGAGGAGT \u00a0552A2allele1 \u00a0GCAGTGTTTGTGTATA-GGGTGGGAGAGAGAGAACAGAGGAGCAGATGGAGGAGT \u00a0542A2allele2 \u00a0GCAGTGTTTG\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014TGGGAGAGAGAGAACAGAGGAGCAGATGGAGGAGT \u00a045\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0BTN2A1\u2212/\u2212 allelic phenotype2) 2A1-49F \u00a0\u00a0\u00a0\u00a0\u00a0GGACTAGGCTCTAAGCCCCTCATTTCAATGAGGGGCCATGAA-GACGGGGGCATCCGGC \u00a059Allele1 \u00a0\u00a0\u00a0\u00a0\u00a0GGACTAGGCTCTAAGCCCCTCATTTCAATGAGGGGCCATGAAGGACGGGGGCATCCGGC \u00a059Allele2 \u00a0\u00a0\u00a0\u00a0\u00a0GGACTAGGCTCTAAGCCCCTCATTTCAATGAGGGG\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014\u2014GGCATCCGGC \u00a045\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0BTN2A2\u2212/\u2212 allelic phenotype3) 2A2 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0CAAGAAGGCAGGTCCTACGATGAGGCCATCCTACGCC-TCGTGGTGGCA \u00a048Allele \u00a0\u00a0\u00a0\u00a0\u00a0CAAGAAGGCAGGTCCTACGATGAGGCCATCCTACGCCCTCGTGGTGGCA \u00a049\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u2217\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0BTN2\u2212/\u2212 cells were seeded overnight in triplicates at 2\u00a0\u00d7 104 cells/well in 100\u00a0\u03bcL DMEM containing 10% FCS in a 96 well flat bottom cell culture plate with or without 25\u00a0\u03bcM Zol. Next day, DMEM with/without Zol was aspirated and cells were washed twice with PBS at RT and 2\u00a0\u00d7 104 expanded V\u03b39V\u03b42 T\u00a0cells/well in 100\u03bcL RPMI was added and cocultured for 4 h. After 4 h, supernatants were collected and frozen at \u221220\u00b0C, until cytokine assay was performed with human IFN\u03b3 ELISA kit (Invitrogen) was used according to the manufacturer\u2019s instructions.293T or 293T BTN2A1 and BTN2A2 were amplified from the cDNA obtained from 293T cells with the help of pMIM-BTN2A1/2Fwd and pMIM-BTN2A1/2Rev primers , V\u03b42-R51A and V\u03b42-CDR3 deletion mutant (\u0394CDR3: CDR3\u03b4 sequence CACD\u2014\u2014\u2013YTDKLIF) TCR chains were generated as reported earlier . V\u03b39-E70Drosophila S2 cells and purified by nickel chromatography as previously described (cDNA encoding wild type BTN2A1 (S27 to V142) or BTN2A2 IgV domains (S31 to V146), or BTN2A1 IgV incorporating the described\u00a0mutations, were generated as gblocks (Integrated DNA Technologies) including the sequence for a C-terminal 6x Histidine\u00a0tag and cloned into the pET23a expression vector (Novagen). Proteins were overexpressed, purified and refolded as described . BTN2A1 escribed . TCRs we5 cells at 4\u00b0C.Flow cytometry staining of the samples were performed with the below mentioned antibodies and samples were measured on FACSCalibur or LSRII flow cytometer (BD). The expression of BTN3A1 and BTN2A1 were detected with anti-huBTN3 (CD277) clone 103.2 (gift from David Olive) and anti-huBTN2A1 clone 1C7D (MBL), followed by secondary antibody F(ab\u2019) Donkey anti mouse IgG\u00a0(H+L) R-PE (Jackson Immunoresearch). mIgG1,\u03ba isotype clone p3.6.2.81 (eBiosciences) and mIgG2a,\u03ba isotype clone-eBM2a (eBiosciences) were used a isotype controls and were detected by above mentioned secondary antibody. N-terminal HA-tagged BTN2A1 or BTN2A2 were detected using anti-HA-FITC (Biolegend). PBMC expanded human V\u03b39V\u03b42 T\u00a0cells were detected with FITC- conjugated anti-human V\u03b42 (BD Biosciences). Biotinylated soluble V\u03b39V\u03b42 TCRs were tetramerized by the addition of Streptavidin-PE conjugate (ThermoFisher Scientific) at room temperature, and 1-2\u03bcg of tetramer used to stain 10SPR was performed as previously described on a BIABTN2A1 and BTN2A2 loci have been functionally inactivated (293T BTN2\u2212/\u2212 cells), or CHO CD80+ cells, were transduced to overexpress HA-tagged BTN2A1 or BTN2A2, or BTN3A1, as indicated. Cells were surface biotinylated using EZ-Link Sulfo NHS-LC-biotin for 30\u00a0min on ice, quenched with 20mM Tris pH 7.5 for 5\u00a0min, washed in TBS, and lysed in lysis buffer containing 1% NP40 in 20mM Tris pH 7.5, 150mM NaCl\u00a0\u00b1 10mM iodoacetamide (Sigma). HA-tagged BTN2A1 or BTN2A2 was immunoprecipitated using anti-HA resin (BioLegend). Immunoprecipitations were washed in lysis buffer and eluted in nonreducing (NR) or reducing (R) SDS sample buffer and boiled, or incubated at 37C for 5\u00a0min before separation on 4%\u201320% SDS-PAGE gels (BioRad). Proteins were transferred to PVDF using the BioRad TransBlot Turbo system, blocked in 3% BSA, then incubated with streptavidin-HRP (Thermo). To investigate potential association of BTN2A1 and BTN3A1 at the cell surface, CHO cells overexpressing BTN2A1-HA, FLAG-BTN3A1, or both, were treated with the soluble, membrane-impermeable crosslinker sulfo-EGS (ThermoFisher) at 0.5mM in PBS, at 4\u00b0C for 2 h. Following this, the reaction was quenched by addition of Tris pH 7.5 to 20mM. Cells were washed in TBS and lysed in 1% NP40 lysis buffer. After centrifugation to remove insoluble material, immunoprecipitation was carried out using anti-HA resin or 20.1 antibody bound to protein A Sepharose . Immunoprecipitates were run on duplicate 4%\u201320% gels (BioRad) and blotted with anti-BTN2A1 or anti-FLAG antibodies.293T cells in which the The ectodomain structures of BTN2A1 (residues Q29-A248) and BTN2A2 (residues Q33-M265), were generated using the I-TASSER (Iterative Threading ASSEmbly Refinement) server . BrieflyThe BTN2A1-IgV/V\u03b39 complex was modeled with HADDOCK . BTN2A1 R65 (located in the IgV domain of BTN2A1) forms a salt bridge interaction with E76 (in the V\u03b39 TCR chain). This interaction is likely to be abolished by introducing Ala at this position in BTN2A1 IgV domain (R65A), consistent with abrogation of binding by the R65A mutant. The hydroxyl group of S72 (BTN2A1) is in close proximity to V58 (TCR). Juxtaposition of this polar residue (S72) with a hydrophobic residue (V58) is likely to be energetically unfavorable for binding in this region. By substituting an Ala (ie a non-polar residue) at this position, the S72A mutation is likely to introduce hydrophobic interactions with V58, consistent with enhanced binding compared to wild-type (11-15\u03bcM (S72A) versus 50\u03bcM (Wild-type)).K79A leads to reduced binding to TCR (100\u03bcM). K79 forms a salt bridge interaction with E76 (TCR). Change to Ala will result in loss of this interaction consistent with a reduction in binding affinity. The fact that binding is not totally abolished suggests that this interaction is a not a major contributor to the binding energy. Note however that E76 also contacts R65 (see above).R124A mutation in BTN2A1 abolishes binding to TCR. The HADDOCK model suggests that R124 forms a salt bridge interaction with E70 (V\u03b39-IgV TCR) and a hydrogen bonding interaction with the hydroxyl group of T83. These interactions will be lost when introducing an Ala at this position.Y126A abolishes binding to TCR. Y126 forms multiple hydrophobic stacking interactions with I74 (HV4 region of Vg9-IgV). In addition, the hydroxyl group of Y126 forms a hydrogen bonding interaction with T77. These will be lost upon alanine substitution.Although Y133A is located at the interface with V\u03b39, it does not mediate interactions with V\u03b39 TCR residues and hence it is unsurprising that substitution to alanine does not affect binding affinity. E135 forms a salt bridge interaction with R20 (TCR). Substitution to Ala will result in a loss of this interaction, and consistent with this, E135A mutation abolishes binding to the TCR.1H-15N-labeled BTN3A1. Experiments were processed using Topspin 3.2 (Bruker). All analysis was performed using CCPN Analysis . SPR data was analyzed in BIAevaluation and Origin 2015 (OriginLab).Stimulation data and transcript visualization in https://doi.org/10.17632/ny6bxn4y9s.1The RNaseq data of radiation hybrid clones filtered for transcribed human genes are available at Mendeley data"} +{"text": "However, in the context of human breast cancer, there is a lack of information for potential phenotypic alterations of this crucial immune cell subset. This is partly due to V\u03b42+ \u03b3\u03b4 T cells scarcity in surgical specimens. To break this deadlock, we assessed V\u03b42+ \u03b3\u03b4 T cell phenotypes using untreated breast cancer patients\u2019 peripheral blood, so-called minimally invasive \u201cliquid biopsy\u201d. We show that circulating V\u03b42+ \u03b3\u03b4 T cell phenotypic alterations are already established at diagnosis and related to tumor progression. Notably, terminally differentiated V\u03b42+ \u03b3\u03b4 T cells expressing canonical markers of replicative senescence and exhaustion were significantly associated with tumor-draining lymph node invasion. Our results shed light on the interest of using liquid biopsy to monitor rare events and support V\u03b42+ \u03b3\u03b4 T cell involvement in breast cancer pathogenesis and progression.V\u03b42+ \u03b3\u03b4 T cells in breast cancer is strongly supported by in vitro and murine preclinical investigations, characterizing them as potent breast tumor cell killers and source of Th1-related cytokines, backing cytotoxic \u03b1\u03b2 T cells. Nonetheless, insights regarding V\u03b42+ \u03b3\u03b4 T cell phenotypic alterations in human breast cancers are still lacking. This paucity of information is partly due to the challenging scarcity of these cells in surgical specimens. \u03b1\u03b2 T cell phenotypic alterations occurring in the tumor bed are detectable in the periphery and correlate with adverse clinical outcomes. Thus, we sought to determine through an exploratory study whether V\u03b42+ \u03b3\u03b4 T cells phenotypic changes can be detected within breast cancer patients\u2019 peripheral blood, along with association with tumor progression. By using mass cytometry, we quantified 130 immune variables from untreated breast cancer patients\u2019 peripheral blood. Supervised analyses and dimensionality reduction algorithms evidenced circulating V\u03b42+ \u03b3\u03b4 T cell phenotypic alterations already established at diagnosis. Foremost, terminally differentiated V\u03b42+ \u03b3\u03b4 T cells displaying phenotypes of exhausted senescent T cells associated with lymph node involvement. Thereby, our results support V\u03b42+ \u03b3\u03b4 T cells implication in breast cancer pathogenesis and progression, besides shedding light on liquid biopsies to monitor surrogate markers of tumor-infiltrating V\u03b42+ \u03b3\u03b4 T cell antitumor activity.The rationale for therapeutic targeting of V\u03b42 Breast cancer (BC) remains the most diagnosed and leading cause of cancer death among women worldwide . AlongsiHowever, exploring antitumor immunity patterns in the tumor bed using resected or biopsied primary or metastatic tumors shows multiple constraints. First, apart from being invasive, tumor biopsies are not always repeatedly feasible and are therefore poorly suitable for longitudinal studies or transfer in clinical routine. Second, genetic and immunological heterogeneity exists between and within each metastasis and the primary tumor, which contributes to the complexity of antitumor immunity pattern study using solid biopsies ,16. Thos+ T cells, \u03b3\u03b4 T cells display pro-inflammatory cytokine production and cytotoxic effector function. As for \u03b1\u03b2 CD8+ T cells, \u03b3\u03b4 T cells can be divided into four functionally distinct subsets reflecting their maturation stages, using a combination of canonical markers known to be differentially expressed during the course of T cell maturation. That is, following antigenic stimulation, CD45RA+ CCR7+ CD27+ CD28+ na\u00efve \u03b3\u03b4 T cells mature to CD45RA\u2212 CCR7+ central memory (CM) \u03b3\u03b4 T cells with low effector function and strong proliferative potential. Still, upon antigenic stimulation, CM \u03b3\u03b4 T cells can further mature to CD45RA\u2212 CCR7\u2212 CD27+/\u2212 CD28+/\u2212 effector memory (EM) \u03b3\u03b4 T cells, producing pro-inflammatory cytokines and cytolytic protein . EM \u03b3\u03b4 T cells finally mature to CD45RA+ CCR7\u2212 CD27\u2212 CD28\u2212 re-expressing CD45RA terminally differentiated effector memory (TEMRA) \u03b3\u03b4 T cells with low proliferative potential and strong cytotoxic function [Human \u03b3\u03b4 T cells represent a unique conserved lineage of T cells, which are responsive against viral/microbial pathogens and transformed cells . Similarfunction ,31,32. W+ \u03b3\u03b4 T cells since they are the predominant subtype of \u03b3\u03b4 T cells in breast tissues. Yet V\u03b41+ \u03b3\u03b4 T cell functions remain controversial, either affiliated with a Th1 or regulatory polarization [+ \u03b3\u03b4 T cells are a potent pro-inflammatory mediator and cytotoxic effectors towards breast tumor cells and have been observed in direct contact with the latter in breast tumors [+ \u03b3\u03b4 T cells are the major subtype of \u03b3\u03b4 T cells in the peripheral blood [+ \u03b3\u03b4 T cells [+ \u03b3\u03b4 T cells phenotypic alterations in human breast cancers and their impact on disease progression is needed.Contrasting from their counterpart \u03b1\u03b2 T cells, \u03b3\u03b4 T cells display a major histocompatibility complex (MHC)-unrestricted antigen presentation and TCR activation, driving their maturation and the expression of natural killer cells (NK) associated cytotoxic receptors. Indeed, \u03b3\u03b4 T cells also differ from \u03b1\u03b2 T cells by expressing natural killer cell receptors such as the NKG2 receptors family. Hence, \u03b3\u03b4 T cells carry hallmarks of innate as well as adaptive immune responses . In brearization ,38,39. Ot tumors ,41,42. Wal blood . Of note T cells ,44,45. H+ \u03b3\u03b4 T cells phenotypic alteration detectable at diagnosis of early-stage breast cancer. Notably, peripheral PD-1+ or CD57+ EMRA V\u03b42+ \u03b3\u03b4 T cells are associated with the pathological involvement of tumor-draining axillary lymph nodes.To this end, we used mass cytometry to quantify up to 130 immune variables from the peripheral blood of untreated breast cancer patients. We evidenced peripheral V\u03b42A total of 122 immune variables were quantified from peripheral blood mononuclear cells (PBMCs) of 13 newly diagnosed BC patients and four healthy volunteers (HV) using two mass cytometry panels Table S1Because there are only two groups, BGA output is a single dimension discriminating axis, where each sample is positioned according to the coexpression of the 122 variables used as input . Still, in the HV group, T cell populations displayed poorly differentiated and non-activated profiles. Indeed, the frequency of Na\u00efve (CD45RA+ CCR7+ CD27+ CD28+) V\u03b42+ \u03b3\u03b4 T cells, Na\u00efve \u03b1\u03b2CD8+ T cells, Na\u00efve \u03b1\u03b2CD4+ conventional T cells, central memory V\u03b42+ \u03b3\u03b4 T cells, resting (CD45RA+ CCR7+ CTLA-4low ICOSlow) Tregs and Tregs expressing a marker associated with a disrupted immunosuppressive activity (DNAM1+), were increased in HV samples compared to BC samples. On the opposite, T cells displaying a differentiated and polarized phenotype were associated with the BC group. Indeed, the frequency of effector memory T cells re-expressing CD45RA V\u03b42+ \u03b3\u03b4 T cells, TEMRA \u03b1\u03b2CD8+, late effector memory \u03b1\u03b2CD8+ T cells and early effector memory \u03b1\u03b2CD4+ conventional T cells were increased in BC samples compared to HV samples. Additionally, an increased frequency of highly cytotoxic (CD8+) NK cells was also detected in BC samples. Importantly, frequencies of V\u03b42+ \u03b3\u03b4 T, \u03b1\u03b2CD8+ T and NK cells expressing inhibitory receptors were also increased in BC samples.In the group of HV, we detected an increased frequency of NK cells expressing NK triggering receptors , 7% (NK NKG2C+), 6.3% (V\u03b42+ \u03b3\u03b4 CD4+), 4.7% (V\u03b42+ \u03b3\u03b4 CM), 4.1% (\u03b1\u03b2CD8+ Naive), 4% (NK IL7R+), 3.5% (Tregs DNAM1+), 3.1%(Tregs resting) and 3% of contribution to the discrimination of groups (+ \u03b3\u03b4 KIR2DL1/DS1+), 6.1% (\u03b1\u03b2CD8+ KIR2DL1/DS1+), 4.5% (V\u03b42+ \u03b3\u03b4 KIR2DL2/DL3+), 4.2% (\u03b1\u03b2CD8+ LEM), 4% , 3.7% (\u03b1\u03b2CD8+ TEMRA), 3.5% (\u03b1\u03b2CD8+ LAG3+) and 3.2% (NK CD8+) of contribution to the discrimination of groups and six BC patients harboring invaded tumor-draining lymph nodes (BC N+). Among the 130 immune variables, 32, 31, 20, 19 and 28 variables were, respectively expressed by V\u03b42ications A.+) highly cytotoxic (CD56+) V\u03b42+ \u03b3\u03b4 T cells profiles. Of note, TEMRA V\u03b42+ \u03b3\u03b4 T cells were the unique population harboring a significantly increased frequency in BC N+ patients compared to BC N\u2212 patients and a canonical marker of T cell functional replicative senescence (CD57+) tends to increase in some patients harboring lymph node invasion, although the difference was not significant. The frequency of NK cells and \u03b1\u03b2CD4+ T cells expressing inhibitory immune checkpoint receptors also tended to increase in some patients harboring lymph node invasion.Interestingly, BC N+ patients displayed fully differentiated (TEMRA), activated (ICOSpatients . However+ \u03b3\u03b4 TEMRA), 7% (V\u03b42+ \u03b3\u03b4 CD57+), 4.5% , 4.1% (V\u03b42+ \u03b3\u03b4 TIM3+), 3.6% , 3.4% (V\u03b42+ \u03b3\u03b4 KIR2DL1/DS1+), 3% (V\u03b42+ \u03b3\u03b4 ICOS+) and 2.7% of contribution to the discrimination of groups (+ \u03b3\u03b4 KIR2DL1/DS1+), 6% (\u03b1\u03b2CD8+ KIR2DL1/DS1+), 5% (V\u03b42+ \u03b3\u03b4 KIR2DL2/DL3+), 4% (\u03b1\u03b2CD8+ LEM) and 3% , 3.7% (\u03b1\u03b2CD8+ TEMRA), 3.5% (\u03b1\u03b2CD8+ LAG3+) of contribution to the discrimination of groups , cytotoxic potential , activation state , replicative senescence (CD57), inhibitory signaling and tumor-promoting tolerance , susceptibility to apoptosis (Fas), tumor cell recognition and costimulatory signaling (DNAM-1) A. Twenty(DNAM-1) . Echoing+ \u03b3\u03b4 T cell clusters that coexpressed inhibitory checkpoint receptors and T cells replicative senescence markers were found to be associated with lymph node involvement (+ PD-L1+ BTLA+ EEM (cluster 2), TIGIT+ CTLA-4+ PD-1+ EEM (cluster 12), CD16+ CD56+ CD57+ PD-1\u2212/+ TIGIT+/\u2212 PD-L1\u2212/+ TEMRA V\u03b42+ \u03b3\u03b4 T cells were exclusively found in patients with lymph node involvement approved by the institutional review board (Comit\u00e9 d\u2019Orientation Strat\u00e9gique (COS), Marseille, France) of the Paoli-Calmettes Institute. Written informed consent was obtained from all patients in concordance with the Declaration of Helsinki. Age-matched healthy subjects were recruited from the Etablissement Fran\u00e7ais du Sang (EFS Alpes-M\u00e9diterran\u00e9e).Breast cancer patients treated at the Institut Paoli-Calmettes were prospectively enrolled between February 2018 and June 2018. Fresh EDTA-anticoagulated blood samples were obtained from patients at diagnosis (n = 13) and healthy volunteers (N = 4). Blood samples were processed extemporaneously. After analysis of morphological tumor characteristics by pathologists, patients were classified as summarized in \u00ae instrument, Fluidigm), at an acquisition rate of approximately 500 events per second. Following the manufacturer\u2019s instructions, settings were on default.PBMCs were obtained using Ficoll-Paque density-gradient centrifugation. Cells were consecutively centrifuged and incubated with cisplatin 0.1 M to stain dead cells. Aspecific epitope binding was blocked with 0.5 mg/mL human Fc Block . The two mass cytometry panels used are provided in + T cells, \u03b1\u03b2CD8+ T cells, \u03b1\u03b2CD4+ conventional T cells, Tregs and NK cells were manually gated according to the gating strategy displayed in TCR\u03b3\u03b42+ \u03b3\u03b4 T cells were exported using FlowJo V10.6.2. Prior to t-SNE analyses using Cytosplore V2.2.1 [+ \u03b3\u03b4 T cells for each file. The t-SNE analysis was carried out with the default setting (perplexity of 30 and 1000 iterations), and V\u03b42+ \u03b3\u03b4 T cell subpopulations were automatically defined by cell density gradient. The results of the t-SNE analysis for each panel are provided in Manually gated V\u03b42e V2.2.1 , consensp-value < 0.05 was considered as significant.Statistical analyses were generated using GraphPad Prism V5.00. Data are expressed as mean \u00b1 standard error of the mean (SEM). Statistical significance between two groups was calculated using the nonparametric Mann\u2013Whitney test. A + \u03b3\u03b4 T cells, phenotypic alterations were evidenced in newly diagnosed breast cancer patients. PD-1+ or CD57+ EMRA V\u03b42+ \u03b3\u03b4 T cells associated with lymph node invasion. These findings evidenced in the context of an exploratory study require confirmation in a larger cohort. However, they open new perspectives for the development of V\u03b42+ \u03b3\u03b4 T cell immunotherapy in breast cancer.Circulating V\u03b42"} +{"text": "Scientific Reports 10.1038/s41598-020-62990-0, published online 20 April 2020Correction to: This Article contains errors in Table 1 where two column headings are incorrect.d13C (\u2030)\u201d and \u201cd15N (\u2030)\u201d.\u201cshould read:\u03b413C (\u2030)\u201d and \u201c\u03b415N (\u2030)\u201d, respectively.\u201c"} +{"text": "RDPBR\u2019 Pd0 and PdII complexes (RDPBR\u2019=(o\u2010PR2C6H4)2BR\u2019, diphosphinoborane) was analyzed using XRD, 11B\u2005NMR spectroscopy and NBO/NLMO calculations. The borane acceptor discriminates between the oxidation state PdII and Pd0, stabilizing the latter. Reaction of lithium amides with [(RDPBR\u2019)PdII(4\u2010NO2C6H4)I] chemoselectively yields the C\u2212N coupling product. DFT modelling indicates no significant impact of PdII\u2192B coordination on the inner\u2010sphere reductive elimination rate.The dative Pd\u2192B interaction in a series of Ligand effects: The borane acceptor in diphosphinoborane ligands discriminates between the oxidation state PdII and Pd0, stabilizing the latter. The impact of the Pd\u2192B interaction on inner\u2010sphere C\u2212N bond reductive elimination of N,N\u2010dimethyl\u20104\u2010nitroaniline was investigated (see scheme). We determine how the oxidation state of Pd and co\u2010ligands affect the strength of the Pd\u2192B interaction in DPB complexes. NBO/NLMO calculations and solid\u2010state structures are used to assess the strength of Pd\u2192B interactions. The value of the 11B\u2005NMR chemical shift as a probe is discussed. The reductive elimination of N,N\u2010dimethyl\u20104\u2010nitroaniline from [(PhDPBPh)PdII(4\u2010NO2\u2010C6H4)NMe2] (5) was studied and modelled with DFT calculations to investigate the assumed influence of the borane acceptor.We speculated that the electron\u2010withdrawing properties of the borane functionality in diphosphinoborane (DPB) ligands enhances the rate of inner\u2010sphere reductive elimination from Pd complexes due to 1)\u2005overall reduced electron density at the PdPhDPBPh)PdII] complexes was synthesized to examine a possible correlation between the nature of ligands at Pd and the strength of the PdII\u2192B interaction (Scheme\u2005A series of [(n Scheme\u2005.PhDPBPh)PdIICl2] (7) was produced by reaction of PhDPBPh ligand with [(cod)PdCl2] in DCM and was isolated in 74\u2009% yield \u2005\u00c5 is shorter than the sum of the van der Waals radii (3.28\u2005\u00c5),2\u2010 type coordination to the PdII center. A slightly increased pyramidalization at the boron atom is observed (\u03a3B\u03b1=355.4\u00b0) compared to complex [(iPrDPBPh)PdCl2] (\u03a3B\u03b1=359.9\u00b0).Complex and was isolated in 67\u2009% yield. The 31P\u2005NMR spectrum displays two broad resonances of equal intensity at \u03b4=45.2 and 38.1\u2005ppm (CD2Cl2), suggesting a similar dynamic process as in 7. Due to the poor solubility of both 7 and 8, no 11B\u2005NMR spectra could be obtained.Complex\u2005PhDPBPh)PdIICl]SbF6 (9) was produced in 51\u2009% isolated yield by halide abstraction from 7 with AgSbF6 PdII(\u03bc\u2010Cl)]2(SbF6)2 is observed with an inversion center between the two PdII centers. Within the dimer, the PdII center is coordinated in a square\u2010pyramidal fashion with the borane located in the apical position. The Pd, B distance in complex\u20059 is 2.721(5)\u2005\u00c5, which is slightly shorter than in [(PhDPBPh)PdIICl2] 7 (2.762(3)\u2005\u00c5). However, pyramidalization of the borane is almost identical (\u03a3B\u03b1=355.8\u00b0). The absence of a relevant \u03b72\u2192PdII interaction is suggested by the long Pd1,C1 distance of 3.338(4) \u00c5. The Pd,B distance and lack of significant pyramidalization at the borane suggest a weak PdII\u2192B interaction, which is in line with a broad resonance in the 11B\u2005NMR spectrum at \u03b4=65\u2005ppm (\u03c91/2=1900\u00b1500\u2005Hz).Cationic complex SbF6 (10) was synthesized by reaction of AgSbF6 with zwitterionic allyl complex [{(o\u2010PPh2C6H4)2B(OAc)Ph}PdII(C3H5)] (4) \u2005\u00c5, which is in line with a minor pyramidalization at the borane center (\u03a3B\u03b1=354.7\u00b0) and a broad 11B\u2005NMR resonance at \u03b4=62\u2005ppm (\u03c91/2=1200\u00b1100\u2005Hz). A large Pd,C22 distance of 3.066(6)\u2005\u00c5 eliminates the possibility of a strong \u03b72\u2192PdII interaction. The \u03b73\u2010coordinated C3H5\u2010ligand is disordered. Using the borane as a reference point, a 39:61 mixture of the exo\u2010 and endo\u2010isomers is observed. A wider P\u2010Pd\u2010P angle of 102.86(5)\u00b0 is realized by a decrease in the twisting of the ligand backbone . The observed disorder of the C3H5\u2010ligand is in good agreement with the observed NMR spectra. In the 31P\u2005NMR spectrum (CD2Cl2), two singlet resonances are observed in a 40:60 ratio (\u03b4=28.1 and 26.9\u2005ppm) and two sets of C3H5\u2010units are detected in the 1H\u2005NMR spectrum. DFT calculations (BP86/def\u2010SV(P)) based on the solid\u2010state structures of endo10\u2010 and exo10\u2010 indicate a small Gibbs free energy preference of \u0394G=0.74\u2005kcal\u2009mol\u22121 for endo10\u2010, predicting a 29:71 ratio at 298\u2005K.Cationic allyl complex [(H5)]SbF6 0 was syno\u2010PPh2C6H2B(OAc)PhII\u2192B interaction on reductive elimination proceeding via an inner\u2010sphere mechanism, complex [(PhDPBPh)PdII(4\u2010NO2\u2010C6H4)I] (5) was reacted with lithium amides. Complex\u20055 was reacted with LiNMe2 (1.1\u2005equiv) at room temperature in [D8]THF and 38.3\u2005ppm (14\u2009%). After a total of 4.5\u2005h, all resonances in the 31P\u2005NMR spectrum disappeared in favor of the singlet at \u03b4=31.1\u2005ppm. 11B\u2005NMR spectroscopy suggested formation of a zero\u2010valent palladium complex by a broad resonance at \u03b4=19\u2005ppm (\u03c91/2=400\u00b1100\u2005Hz). The concurrent formation of the expected reductive elimination product N,N\u2010dimethyl\u20104\u2010nitroaniline was confirmed by GC/MS analysis, using an independently prepared sample as a reference. The absence of an intermediate complex cis\u2010[(PhDPBPh)PdII(4\u2010NO2\u2010C6H4)NMe2] suggests that transmetalation is rate\u2010limiting in this transformation. The intermediate occurrence of the 31P\u2005NMR resonance at \u03b4=38.3\u2005ppm is possibly due to a reversible reaction of LiNMe2 with complex\u20056. In a control experiment complex [(PhDPBPh)Pd0(pyridine)] (1) was reacted with LiNCy2 and LiNMe2 in [D8]THF. In both cases ca. 7\u2009% of a new complex at \u03b4=38.5 (s) and 37.7\u2005ppm (s) were observed.A conversion of 84\u2009% was observed 6 decomposed within hours with simultaneous precipitation of palladium black. Addition of PMe3 as a stabilizing co\u2010ligand led to the formation of complex [(PhDPBPh)Pd0(PMe3)] 11. The 31P\u2005NMR spectrum of 11 showed a doublet at \u03b4=35.3 and a triplet at \u221240.1\u2005ppm (J=15.1\u2005Hz) in a 2:1 ratio, which is consistent with the expected \u03ba3P\u2010coordination. The broad resonance in the 11B\u2005NMR spectrum at \u03b4=25\u2005ppm (\u03c91/2=400\u00b1100\u2005Hz) suggested a strong Pd0\u2192B interaction. Complex\u200511 could also be synthesized independently by reaction of PBP pincer 12 with PhLi and PMe3, or reaction of 1 with PMe3, thus confirming unambiguously the identity of 11 (Scheme\u2005Complex\u20055 reacted in a similar fashion with LiNCy2 (26\u2009% 6 after 3\u2005h) and LiNHtBu (14\u2009% 6 after 5.5\u2005h). However, the reaction proceeded slower with these sterically more demanding substrates. The reaction of complex\u20055 with LiNHtBu was monitored for 96\u2005h by 31P\u2005NMR spectroscopy (46\u2009% conversion towards 6) without any side products being observed (cf. Table\u2005S1). This is in line with the assumption of a rate\u2010determining transmetalation followed by a quick reductive elimination.Complex\u20050/II DPB complexes were analyzed to identify factors which affect the strength of Pd\u2192B interactions. In addition to the new Pd complexes presented in this work (6\u201310), the structurally characterized DPB complexes cis\u2010[(PhDPBPh)PdII(4\u2010NO2\u2010C6H4)I] (5),PhDPBPh)Pd0(pyridine)] (1),PhDPBMe)Pd0(PMe3)] (13)CyDPBPh)Pd0] (3)0/II center. The shorter Pd,B distances and higher degree of borane pyramidalization =2.194(3) \u22122.243(2)\u2005\u00c5 vs. PdII: \u03a3B\u03b1=354\u2013356\u00b0, d=2.676(5) \u22122.762(2)\u2005\u00c5). Remarkably, even the generation of cationic PdII complexes (9 and 10) has no significant impact on the strength of PdII\u2192B interactions. The oxidation state at Pd is unambiguously the dominant factor for the strength of the Pd,B bond.The solid\u2010state structures of PdPBPh)PdII\u2010NO2\u2010C6H4s work 6\u20130, the st1, 3, 5\u201311 and 13 were geometrically optimized using Turbomole 7.0.1 (BP86/def\u2010SV(P)). A good agreement was observed between the optimized structures and their corresponding solid\u2010state structures \u2010coordination was found in the NBO calculations. The Wiberg bond index for Pd,Cipso was below 0.02, with the exception of Pd0 complexes\u20051 (0.0697) and 6 (0.0325). Reactivity studies of [(DPB)Pd]\u2010complexes presented in this paper thus appear to be unaffected from significant \u03b72\u2010coordination.The Pd\u2192B interactions were further analyzed using QM calculations. Complexes\u2005II\u2192B interactions, a narrow range of NBO stabilizing energies between 8.04 and 11.46\u2005kcal\u2009mol\u22121 was observed. Surprisingly, generation of cationic complexes , exchange of chloro\u2010ligands by bromide (8) or iodide/aryl (5) had very little effect. In the case of Pd0\u2192B interactions, significantly higher NBO stabilizing energies of 19.53\u201346.83\u2005kcal\u2009mol\u22121 were found. Regardless of the oxidation state at Pd an approximately linear correlation between the Pd,B distance and the NBO stabilizing energy (E2) associated with the Pd,B interaction was observed by PCy2\u2010groups (3) had only a minor effect. The E2 values for the Pd0\u2192B interaction in the 14 VE complexes\u20053 and 6 significantly deviate from this correlation and are almost twice as much as for 16\u2005VE complexes\u20051 and 13 . Neither the 11B\u2005NMR chemical shift, Pd,B distance or pyramidalization at B indicate a change of the Pd0\u2192B interaction strength in this magnitude between the 14\u2005VE and the 16\u2005VE complexes Pd0(PPh3)] (2) reported by Kameo and Bourissou\u03b4(11B) 27\u2005ppm). In contrast, the 11B\u2005NMR resonance shifts linearly towards lower field with an increasing Pd,B distance in case of PdII complexes. 11B\u2005NMR spectroscopy therefore can be used as a tool to assess the strength of Pd\u2192B interactions within a given ligand system, provided that the oxidation state at the Pd center is taken into account. However, given the difficulty to determine the precise \u03b4(11B) of [(DPB)PdII] complexes (poor solubility and \u03c91/2 >1000\u2005Hz ), a certain error for weak PdII\u2192B interactions needs to be factored in.The r Figure\u2005. ComplexN,N\u2010dimethyl\u20104\u2010nitroaniline from complex\u200514\u2010B , yielding Pd0 complex\u20056 and N,N\u2010dimethyl\u20104\u2010nitroaniline . In order to understand how the PdII\u2192B interaction affects the reductive elimination, the reaction was also modeled for bis[(2\u2010diphenylphosphino)phenyl]ether (DPEphos) complex\u200514\u2010O and diphosphinoamine complex\u200514\u2010N. DPEphos is well established as an effective ligand in palladium catalyzed Buchwald\u2013Hartwig\u2010type coupling reactions,PhDPBPh (Table\u2005o\u2010PPh2C6H4)2NPhN\u2010Ph bridgehead gives a good model of the B\u2010Ph group in 14\u2010B. Elimination of N,N\u2010dimethyl\u20104\u2010nitroaniline from complexes\u200514\u2010O and 14\u2010N gave very similar Gibbs free reaction energies of \u0394G=\u221238.52\u2005kcal\u2009mol\u22121 and \u0394G=\u221238.63\u2005kcal\u2009mol\u22121, respectively. No Pd0/II\u2192E interactions were observed in complexes featuring DPEphos and the diphosphinoamine ligand \u2005an electronic effect by Pd\u2192B coordination and 2)\u2005increased steric bulk of the DPB ligand imposed by the B\u2010Ph group. In case of diphosphinoamine complex 14\u2010N the reductive elimination barrier decreased to \u0394G\u2260=5.54\u2005kcal\u2009mol\u22121 , possibly as a result of the increased steric pressure imposed by the N\u2010Ph group proceeds via structurally early transition\u2010state 15\u2010E were used to model the inner\u2010sphere reductive elimination of 15\u2010B, compared to starting complex\u200514\u2010B, as indicated by a slightly elongated Pd,B distance (2.947\u2005\u00c5) in 15\u2010B compared to 14\u2010B (2.906\u2005\u00c5). Similarly, the Wiberg bond index for the Pd\u2192B interaction is reduced to 0.162 in 15\u2010B (14\u2010B: 0.176), and the NPA charge at the borane remains unchanged (14\u2010B: +0.737 vs. 15\u2010B: +0.735). The increase of the Pd\u2192B interaction strength occurs after the reductive elimination, explaining why the inner\u2010sphere reductive elimination of the C\u2212N bond does not kinetically profit from the substantial increase of the Pd\u2192B strength in the course of the reaction.Unexpectedly, the Pd\u2192B interaction is slightly weakened in transition\u2010state N,N\u2010dimethyl\u20104\u2010nitroaniline was also modeled using cis\u2010[(PMe3)2PdII(4\u2010NO2C6H4)NMe2] and its BH3 adduct [(PMe3)2(BH3)PdII(4\u2010NO2C6H4)NMe2] as substrates (cf. Scheme\u2005S1). Again, a more favorable transition state was found for the acceptor free complex 17 , than for the borane adduct 17\u2010B .To rule out effects originating from restraints imposed by a chelating ligand frame work, the reductive elimination of 11B\u2005NMR spectroscopy has been established as a useful tool to assess the strength of Pd\u2192B interactions in solution. Reaction of lithium amides with [(PhDPBPh)PdII(4\u2010NO2C6H4)I] (5) chemoselectively yields the C\u2010N coupling product and [(PhDPBPh)Pd0] (6). Inner\u2010sphere reductive C\u2212N bond elimination was modelled with DFT methods for the PhDPBPh ligand. In contrast to reports on acceptor promoted outer\u2010sphere reductive C\u2212N bond elimination,The strength of Pd\u2192B interactions in [(DPB)Pd] complexes depends primarily on the oxidation state of Pd. In contrast, modifications of the DPB ligand or co\u2010ligands have only a minor effect. n\u2010hexane were dried over sodium, distilled under argon prior to use and stored over activated molecular sieves (4\u2005\u00c5).All manipulations were performed under an argon atmosphere using standard Schlenk line and glovebox techniques. Glassware was oven dried at 120\u2009\u00b0C overnight and dried with a heat gun under vacuum prior to use. Tetrahydrofuran was dried by an MBraun solvent purification system. Benzene and 2Cl2 and C6D6 were degassed employing the freeze\u2010pump\u2010thaw technique and stored over activated molecular sieves (4\u2005\u00c5). [D8]THF was dried over activated molecular sieves (3\u2005\u00c5), distilled under an argon atmosphere and degassed employing the freeze\u2010pump\u2010thaw technique. PhDPBPh, [(PhDPBPhOAc)Pd(C3H5)] (4), [(PhDPBPh)Pd(4\u2010NO2C6H4)I] (5) and [{(o\u2010PPh2C6H4)2BPh}PdI] (12) were synthesized according to published procedures.CD\u00ae quick pressure valve NMR tubes. 1H, 11B{1H}, 13C{1H}, 19F{1H}, and 31P{1H} NMR spectra were recorded on a Bruker Avance II or a Bruker Avance spectrometer. 1H and 13C{1H} NMR spectra were referenced to residual solvent resonances as implemented in MesReNova 10.0.2. Infrared spectra were recorded on an Avatar 360 FT\u2010IR E.S.P. device by Nicolet. CHN combustion analysis were carried out on an Elementar EL device by Elementar Analysesysteme GmbH.NMR\u2010experiments were performed in Wilmad7), 1987625 (9) and 1987626 (10) Deposition Number(s) 1987620 ( contain(s) the supplementary crystallographic data for this paper. These data are provided free of charge by the joint Cambridge Crystallographic Data Centre and Fachinformationszentrum Karlsruhe Access Structures service www.ccdc.cam.ac.uk/structures.8]THF (0.25\u2005mL) was added dropwise over a period of 4\u2005min to a stirred solution of nitroarene complex\u20055 in [D8]THF (0.25\u2005mL). The resulting mixture was stirred for another 5\u2005min and then transferred into an NMR tube. Reductive elimination was monitored by 31P\u2005NMR spectroscopy.A solution of the respective lithium amide in [D2Cl2 (8\u2005mL) was added to a mixture of PhDPBPh and [(cod)PdCl2] . The mixture was stirred for 30\u2005min at room temperature. Yellow crystals were formed by overlaying the solution n\u2010pentane (16\u2005mL). Single crystals suitable for X\u2010ray diffraction were grown from a solution of [(cod)PdCl2] and PhDPBPh in CD2Cl2 (0.7\u2005mL) overlaid with benzene (0.3\u2005mL). 11B and 13C\u2005NMR data have not been collected due to poor solubility. 1H\u2005NMR : \u03b4 7.81\u20137.76 , 7.55 , 7.50\u20137.46 , 7.46\u20137.38 , 7.35\u20137.14 , 6.97\u20136.78 , 5.32 . 31P{1H} NMR : \u03b4 44.5 . IR (KBr): \u03bd\u02dc=3643\u20103284 (w), 3049 (w), 1587 (w), 1497 (m), 1433 , 1223 (s), 1158 (vw), 1128 (w), 1093 (vs.), 987 (w), 889 (vw), 864 (vw), 754 (s), 744 (s), 733 (m), 688 (vs.), 667 (w), 611 (m), 600 (s), 542 (m), 523 (vs.), 505 (m) cm\u22121. Elemental analysis calcd (%) for C42H33BCl2P2Pd\u22c5CH2Cl2: C 59.18, H 4.04, found: C 59.61, H 4.33.CHPhDPBPh ligand and [(cod)PdBr2] were solved in DCM (10\u2005mL) and stirred at r.t. for 30\u2005min. The solution was overlaid with n\u2010hexane (20\u2005mL) yielding title compound\u20058 as orange crystals . 11B and 13C\u2005NMR data have not been collected due to poor solubility. 1H\u2005NMR : \u03b4 7.85\u20137.76 , 7.59\u20137.19 . 31P{1H} NMR : \u03b4 45.2 , 38.1 . IR (KBr): \u03bd\u02dc=3424 (s), 3048 (m), 1621 (w), 1587 (w), 1478 (m), 1455 (w), 1432 (s), 1311 (w), 1237 (w), 1220 (s), 1205 (m), 1187 (m), 1153 (w), 1126 (m), 1092 (s), 1027 (w), 1000 (m), 887 (w), 863 (w), 753 (s), 741 (s), 713 (m), 699 (s), 690 (s), 667 (m), 610 (s), 600 (s), 539 (s), 522 (s), 505 (s), 465 (m) cm\u22121. Elemental analysis calcd (%) for C42H33BBr2P2Pd\u22c50.25CH2Cl2: C 56.51; H 3.76, found: C 56.72, H 3.83.The 7 and AgSbF6 were stirred in DCM (15\u2005mL) for 40 minutes. The suspension was filtered through a syringe filter . The clear solution was overlaid with n\u2010hexane (30\u2005mL) yielding the title compound\u20059 as long colorless needles . 1H\u2005NMR : \u03b4 7.97\u20137.92 , 7.80 , 7.69 , 7.65 , 7.55 , 7.47\u20137.34 , 7.27\u20137.16 , 7.00 , 6.83 . 11B{1H} NMR : \u03b4=65 . 13C{1H} NMR : \u03b4=\u03b4 141.79, 135.43 , 134.88 , 134.25, 133.69 , 133.22 , 132.49 , 129.67 , 129.33\u2013128.82 (m), 128.10, 127.13, 126.74, 126.16. 31P{1H} NMR : \u03b4 49.9 . IR (KBr): \u03bd\u02dc=3441 (s), 3058 (w), 1588 (w), 1482 (w), 1435 (s), 1230 (m), 1200 (w), 1125 (w), 1034 (m), 1001 (w), 867 (vw), 752 (s), 702 (s), 692 (s), 659 (vs.), 614 (m), 538 (s), 517 (s), 697 (w) cm\u22121. Elemental analysis calcd (%) for C42H33BClF6P2PdSb\u22c50.25 C6H14: C 51.75, H 3.64, found: C 51.77, H 3.785.Complex\u20054 and AgSbF6 were solved in CH2Cl2 (7\u2005mL) and stirred at r.t. for 20\u2005min. The suspension was filtered through a syringe filter . The clear solution was overlaid with n\u2010hexane (10\u2005mL). The obtained crystals showed insufficient purity and were crystallized again under the same conditions yielding 10 as slightly yellow crystals . 1H\u2005NMR : \u03b4 7.72\u20137.59 , 7.58\u20137.53 , 7.53\u20137.44 , 7.43\u20137.29 , 7.23\u20137.15 , 7.05\u20136.87 , 6.78\u20136.67 , 5.88\u20135.70 , 3.77\u20133.61 , 3.59\u20133.33 , 3.03\u20132.85 , 2.49\u20132.29 . 11B{1H} NMR : \u03b4 64 . 13C{1H} NMR : \u03b4 141.1, 140.2, 136.1, 135.5, 135.3, 135.0, 134.4, 134.3, 134.0, 133.2 , 132.3, 132.2, 132.1, 131.6, 131.5, 131.2, 131.0, 129.6 , 129.3, 128.9, 123.1, 80.4, 80.2. 31P{1H} NMR : \u03b4 28.1 , 26.9 . IR (KBr): \u03bd\u02dc=3430 (s), 3000 (m), 1588 (m), 1480 (m), 1458 (w), 1434 (s), 1268 (m), 1227 (s), 1127 (m), 1095 (m), 1031 (w), 999 (w), 950 (vw), 875 (w), 772 (w), 754 (m), 742 (m), 733 (m), 695 (s), 659 (vs.), 609 (s), 537 (m), 521 (s), 478 (w), 430 (w) cm\u22121. Elemental analysis calcd (%) for C46H40BCl2F6P2PdSb: C 51.22, H 3.74, found: C 51.04, H, 3.86.Allyl complex\u20052\u22c5THF in [D8]THF (0.25\u2005mL) was added over a period of 3\u2005min to a solution of complex\u20055 in [D8]THF (0.25\u2005mL). The combined solutions were transferred to an NMR tube and NMR spectra were recorded after 1.5 and 4.5\u2005h. 11B{1H} NMR : \u03b4 19 . 31P{1H} NMR : \u03b4 30.93 (s).A solution of LiNMe12 in THF (0.5\u2005mL). After stirring for 10\u2005min at r.t. a solution of PMe3 in toluene was added. The precipitate was removed by filtration and the solution was concentrated in vacuo. The resulting solid was washed with pentane and dried in vacuo . 1H\u2005NMR : \u03b4 8.34 , 7.69\u20137.58 , 7.44\u20137.37 , 7.36\u20137.28 , 7.12 , 7.09\u20137.05 , 6.85 , 6.68 , 0.64 . 11B{1H} NMR : \u03b4 25 . 13C{1H} NMR : \u03b4 168.7 (bs), 143.2 , 143.0 , 141.5 , 138.9 , 135.8 , 135.7 , 133.5 , 133.0 , 132.3 (s), 132.3 (s), 132.4 , 129.5 (s), 129.0 (s), 128.6 (s), 127.2 (s), 126.1 , 125.2 (s), 18.1 . 31P{1H} NMR : \u03b4 35.44 , \u221240.13 .A solution of PhLi in THF (0.5\u2005mL) was slowly added to a solution of complex\u2005The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "The title hydrazine carbodi\u00adthio\u00adate chloro\u00adform hemi-solvate features approximately planar mol\u00adecules with splayed phenyl groups. Supra\u00admolecular tapes are formed in the crystal through a combination of N\u2014H\u22efS, hydroxyl-O\u2014H\u22efO(hydrox\u00adyl) and hydroxyl-O\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adactions. 15H14N2O2S2\u00b7CHCl3, comprises two independent hydrazine carbodi\u00adthio\u00adate mol\u00adecules, A and B, and a chloro\u00adform mol\u00adecule; the latter is statistically disordered about its mol\u00adecular threefold axis. The common features of the organic mol\u00adecules include an almost planar, central CN2S2 chromophore [r.m.s. deviation = 0.0203\u2005\u00c5 (A) and 0.0080\u2005\u00c5 (B)], an E configuration about the imine bond and an intra\u00admolecular hydroxyl-O\u2014H\u22efN(imine) hydrogen bond. The major conformational difference between the mol\u00adecules is seen in the relative dispositions of the phenyl rings as indicated by the values of the dihedral angles between the central plane and phenyl ring of 71.21\u2005(6)\u00b0 (A) and 54.73\u2005(7)\u00b0 (B). Finally, a difference is seen in the disposition of the outer hydroxyl-H atoms, having opposite relative orientations. In the calculated gas-phase structure, the entire mol\u00adecule is planar with the exception of the perpendicular phenyl ring. In the mol\u00adecular packing, the A and B mol\u00adecules assemble into a two-mol\u00adecule aggregate via N\u2014H\u22efS hydrogen bonds and eight-membered {\u22efHNCS}2 synthons. The dimeric assemblies are connected into supra\u00admolecular chains via hydroxyl-O\u2014H\u22efO(hydrox\u00adyl) hydrogen bonds and these are linked into a double-chain through hy\u00addroxy-O\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adactions. The double-chains are connected into a three-dimensional architecture through phenyl-C\u2014H\u22efO(hydrox\u00adyl) and phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adactions. The overall assembly defines columns along the a-axis direction in which reside the chloro\u00adform mol\u00adecules, which are stabilized by chloro\u00adform\u2013methine-C\u2014H\u22efS(thione) and phenyl-C\u2014H\u22efCl contacts. The analysis of the calculated Hirshfeld surfaces, non-covalent inter\u00adaction plots and inter\u00adaction energies confirm the importance of the above-mentioned inter\u00adactions, but also of cooperative, non-standard inter\u00adactions such as \u03c0(benzene)\u22ef\u03c0(hydrogen-bond-mediated-ring) contacts.The title hydrazine carbodi\u00adthio\u00adate chloro\u00adform hemisolvate, 2C Di\u00adthio\u00adcarbazato Schiff bases have received considerable attention because of the presence of both soft sulfur and hard nitro\u00adgen atoms As part of on-going studies in this area \u2005\u00c5, for the N2 atom, and 0.0319\u2005(16)\u2005\u00c5 for N1; the C2 and C9 atoms lie, respectively, 0.161\u2005(3) and 0.096\u2005(4)\u2005\u00c5 out of the plane, in the direction of the N2 atom. The comparable plane for the S3-mol\u00adecule is significantly more planar with an r.m.s. deviation = 0.0080\u2005\u00c5 with maximum deviations of 0.0131\u2005(16)\u2005\u00c5 for the N3 atom and 0.0104\u2005(12)\u2005\u00c5 for atom N4; the C17 atom lies 0.018\u2005(3)\u2005\u00c5 out of the central plane in the direction of the N3 atom, and the C24 lies 0.123\u2005(3)\u2005\u00c5 out of the plane in the direction of the N4 atom. The small difference in planarity is reflected in the C1\u2014N1\u2014N2\u2014C2 and C16\u2014N3\u2014N4\u2014C17 torsion angles of 171.8\u2005(2) and 179.3\u2005(2)\u00b0, respectively. More significant conformational differences are apparent in rest of the mol\u00adecules: for the S1-mol\u00adecule, the dihedral angles between the central residue and terminal hy\u00addroxy\u00adbenzene and phenyl rings are 6.18\u2005(13) and 77.21\u2005(6)\u00b0, respectively, indicating close to co-planar and perpendicular relationships; the dihedral angle between the terminal rings is 71.22\u2005(8)\u00b0. The equivalent dihedral angles for the S3-mol\u00adecule are 6.07\u2005(13), 54.53\u2005(6) and 54.73\u2005(7)\u00b0, respectively. The other notable difference between the mol\u00adecules relates to the relative orientation of the hy\u00addroxy-H atoms in the 4-position, no doubt arising owing to the dictates of the mol\u00adecular packing.The crystallographic asymmetric unit of (I)E in each case. The comparison of geometric parameters in Table\u00a01The relatively co-planar relationship between the central residue and the appended hy\u00addroxy\u00adbenzene ring allows for the formation of an intra\u00admolecular hy\u00addroxy-O\u2014H\u22efN(imine) hydrogen bond in each mol\u00adecule, Table\u00a02Gaussian16 An overlay diagram for the experimental and theoretical, gas-phase structures is shown in Fig.\u00a02via O4-hy\u00addroxy-O\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adactions as illustrated in Fig.\u00a03a). The assembly lies parallel to more than \u22120.18 a.u., Fig.\u00a06b) and (c). The intra\u00admolecular O\u2014H\u22efN contacts reveal similar attractive inter\u00adactions.Among all close contacts present in (I)NCIPLOT results, the strength of inter\u00adaction for each close contact was qu\u00adanti\u00adfied by calculation of the inter\u00adaction energy in Gaussian16 , the combination of \u03c0(C3\u2013C8)\u2013quasi-\u03c0, quasi-\u03c0\u2013quasi-\u03c0, C24\u2014H24A\u22efS2, C14\u2014H14\u22ef \u03c0(C25\u2013C30) and C15\u2014H15\u22ef\u03c0(C25\u2013C30) between S1- and S3-mol\u00adecules exhibits the greatest inter\u00adaction energy among all close contacts with an E of \u221265.73\u2005kJ\u2005mol\u22121, Table\u00a052 synthon, being the second strongest inter\u00adaction with E = \u221259.79\u2005kJ\u2005mol\u22121. The strength of the N1\u2014H1N\u22efS3/N3\u2013H3N\u22efS1 inter\u00adaction is consistent with the energy range of \u221254.06 to \u221257.99\u2005kJ\u2005mol\u22121 displayed by the equivalent contacts in the cinnamaldehyde Schiff base of S-(4-methyl\u00adbenz\u00adyl) di\u00adthio\u00adcarbaza\u00adtes calculated through wB97XD/6-31G C=NN(H)C(=S)SR, five of which have the hydroxyl substituent in the 2-position enabling the formation of an intra\u00admolecular hy\u00addroxy-O\u2014H\u22efN(imine) hydrogen bond. In the most closely related compounds, i.e. with 2-OH substituents, the R group in the ester substituent is methyl and 2,4-di\u00adhydroxy\u00adbenzaldehyde were mixed and heated until the initial volume was reduced by half. The yellow precipitate formed after cooling the mixture to room temperature was collected and washed with cold ethanol. It was recrystallized from ethanol solution and dried over silica gel for three days. Light-yellow prisms were obtained from its 1:1 diethyl ether/chloro\u00adform solution by slow evaporation.Two solutions, \u22121): 3310 , 3094 , 1604 , 1100 , 1024 , 948 . 1H NMR : \u03b4 13.19 , 10.21 , 10.07 , 8.35 , 7.45 , 7.36 , 7.21 , 6.29 , 6.26 , 4.44 . 13C{1H}-NMR : \u03b4 ppm. 194.4 (C=S), 162.1, 159.6 (C\u2014OH), 146.5 (N=C), 129.7\u2013127.8 (Ph & benzene), 38.0 (CH2); GCMS (DI): m/z calculated for C15H14N2O2S2+ [M+]: 318, found 318.Yield: 4.98\u2005g, 62%; m.p. 463\u2013465\u2005K. FT\u2013IR UATR (solid), \u03bb (cmUiso(H) set to 1.2Ueq(C). The O- and N-bound H atoms were located in a difference-Fourier map but, were refined with O\u2014H (0.84\u00b10.01\u2005\u00c5) and N\u2014H (0.88\u00b10.01\u2005\u00c5) distance restraints, and with Uiso(H) set to 1.5Ueq(O) and to 1.2Ueq(N), respectively. The CHCl3 solvent mol\u00adecule is statistically disordered about the mol\u00adecular threefold axis. The C31 atom is common to both conformations and the individual Cl atoms were refined anisotropically. A loose distance restraint for C\u2014Cl was applied, i.e. C\u2014Cl = 1.76\u00b10.02\u2005\u00c5. The maximum and minimum residual electron density peaks of 1.04 and 1.22\u2005e\u2005\u00c5\u22123, respectively, are located 1.03 and 0.90\u2005\u00c5 from the Cl3\u2032 atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989020007070/hb7919sup1.cifCrystal structure: contains datablock(s) . DOI: 10.1107/S2056989020007070/hb7919Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020007070/hb7919Isup3.cmlSupporting information file. DOI: 2005815CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The cation and anion are linked together by supra\u00admolecular inter\u00adactions [graph-set notation of hydrogen bonding a-axis direction]. Additionally, there is \u03c0\u2013\u03c0 stacking involving the salicylate anion and the piperazinium cation in adjacent asymmetric units as well as a C\u2014H\u22ef\u03c0 inter\u00adaction between a hydrogen atom on the piperazine ring and the phenyl ring within the salicyclate anion. In bis\u00ad[4-(4-nitro\u00adphen\u00adyl)piperazin-1-ium] bis\u00ad(4-fluoro\u00adbenzoate) trihydrate (2C10H14N3O2+\u00b72C7H4FO2\u2212\u00b73H2O), there are two cations, two anions, and three water mol\u00adecules of solvation in the asymmetric unit, all linked by hydrogen bonds [graph-set notation of hydrogen bonding R22(20) between adjacent cations and R33(9) between a cation and its adjacent anion]. In the anion, the 4-nitro\u00adphenyl ring occupies an axial substitution position in the piperazinium ring, which is relatively rare. Within the asymmetric unit, the phenyl groups in the cations show an offset \u03c0\u2013\u03c0 inter\u00adaction. Additionally, there is a C\u2014H\u22ef\u03c0 inter\u00adaction between a hydrogen atom on the phenyl ring within a cation and the phenyl ring within an anion. In 4-(4-nitro\u00adphen\u00adyl)piperazin-1-ium 3,5-di\u00adnitro\u00adbenzoate (C10H14N3O2+\u00b7C7H4N2O6\u2212), there is a strong N\u2014H\u22efO hydrogen bond linking the cation and anion and the 4-nitro\u00adphenyl ring occupies an axial substitution position in the piperazinium ring, as seen in the previous structure. In the crystal, the cation and the anion form a complex three-dimensional hydrogen-bonded array involving R22(8), R44(12) and R44(20) rings propogating in the a-axis direction. The nitro\u00adphenyl group is disordered with occupancies of 0.806\u2005(10) and 0.194\u2005(10).The structures and Hirshfeld surface analysis of three salts of 1-(4-nitro\u00adphenyl)\u00adpiperazine are discussed. In 4-(4-nitro\u00adphen\u00adyl)piperazin-1-ium salicylate (C Additionally, ring B\u2032s piperazine substituent forms a weak C\u2014H\u22efO inter\u00adaction with an oxygen atom in ring C\u2032s terminal nitro group . Ring C\u2032s piperazine substituent forms a similar inter\u00adaction with ring B\u2032s terminal nitro group . In the conformation of rings A\u2013D, the dihedral angles between the 4-nitro\u00adphenyl rings in rings B and C, the 4-nitro\u00adphenyl ring and nitro group in ring B, the 4-nitro\u00adphenyl ring and nitro group in ring C, the piperazine ring and the 4-nitro\u00adphenyl ring in ring B, the piperazine ring and 4-nitro\u00adphenyl ring in ring C, the fluoro\u00adbenzene ring in ring D and the phenyl ring in ring C, and the fluoro\u00adbenzene ring in A and the phenyl ring in C are 11.4\u2005(4), 1.1\u2005(2), 0.2\u2005(2), 141.72\u2005(16), 145.17\u2005(17), 101.47\u2005(17) and 103.32\u2005(17)\u00b0, respectively. The third and fourth angles listed indicate that the 4-nitro\u00adphenyl ring occupies an axial position in both cations, which is relatively rare. In a previous paper containing eleven analogous structures, only one had this substitution pattern , crystallizes in the monoclinic space group C2/c with eight formula units in the unit cell. The structure consists of a 4-nitro\u00adphenyl\u00adpiperazinium cation and a 3,5-di\u00adnitro\u00adbenzoate anion linked by a strong N\u2014H\u22efO hydrogen bond . Additionally, there is a C\u2014H\u22ef\u03c0 inter\u00adaction between a hydrogen atom in the piperazine ring and the phenyl ring within the salicyclate anion x, y, z).In discussing the supra\u00admolecular features of the three structures, the direct hydrogen bonding involving the linking of the 4-nitro\u00adphenyl\u00adpiperazinium cations and organic acid anions is omitted since it has already been discussed in the previous section. For 2, there are two anions and two cations as well as three water mol\u00adecules of solvation in the asymmetric unit. This leads to a complex three-dimensional array of hydrogen bonding involving both B and C, and R33(9) motifs between rings C and D as well as one water mol\u00adecule, as seen in Fig.\u00a06x, y, \u2212z; see Table\u00a02B forms an offset \u03c0\u2013\u03c0 inter\u00adaction with the phenyl ring in C . Additionally, there is a C\u2013H\u22ef\u03c0 inter\u00adaction between a hydrogen atom on the phenyl ring in C and the phenyl ring in D .For 3, the cation and the anion form a complex three-dimensional array of hydrogen bonding involving a-axis direction between the 4-nitro\u00adphenyl group of one cation with the piperazinium ring of an adjacent cation , two cations and two anions in adjacent asymmetric units , two cations and two anions in adjacent asymmetric units , respectively, as seen in Fig.\u00a09Cg2\u22efCg2i distance, 4.4132\u2005(9)\u2005\u00c5; perpendicular distance: 3.5596\u2005(9)\u2005\u00c5; slippage of 2.609\u2005\u00c5, symmetry code: (i) x, y, 1\u00a0\u2212\u00a0z; Cg2 is the centroid of the C1\u2013C6D ring].For 4.et al., 1997et al., 2014et al., 2019Related structures containing 1-phenyl\u00adpiperazine or the 1-phenyl\u00adpiperazinium cation include racemic perhydro\u00adtri\u00adphenyl\u00adene (PHTP), which has been shown to form a polar inclusion compound with 1-(4-nitro\u00adphen\u00adyl)piperazine (NPP) as a guest mol\u00adecule -4-(4-nitro\u00adphen\u00adyl)piperazine (BIQYIM), 1-(4-bromo\u00adbenzo\u00adyl)-4-(4-nitro\u00adphen\u00adyl)piperazine (BIRHES), 1-(3-bromo\u00adbenzo\u00adyl)-4-(4-nitro\u00adphen\u00adyl)piperazine (BIRHIW) and bis\u00ad[(4-fluoro\u00adphen\u00adyl)methanone] (BIRGOB) have been reported 4-nitro\u00adphenyl\u00adpiperazine in methanol (10\u2005ml) was mixed with equimolar solutions of the appropriate acids in methanol (10\u2005ml) and ethyl acetate (10\u2005ml) viz., salicylic acid (67\u2005mg) for 1, 4-fluoro\u00adbenzoic acid (68\u2005mg) for 2 and 3,5-di\u00adnitro\u00adbenzoic acid (102\u2005mg) for 3 for a week. For 3, DMF (3\u2005ml) was used for crystallization. The corresponding melting points were 453\u2013458\u2005K (1), 373\u2013378\u2005K (2) and 445\u2013447\u2005K (3).For the synthesis of salts 6.Uiso(H) = 1.2Ueq(C) while the N\u2013H and water O\u2013H hydrogen atoms were refined isotropically. In 3 the nitro\u00adphenyl group is disordered with occupancies of 0.806\u2005(10)/0.194\u2005(10) and constrained to have similar metrical parameters.Crystal data, data collection and structure refinement details for the three structures are summarized in Table\u00a0410.1107/S2056989023002517/vm2276sup1.cifCrystal structure: contains datablock(s) 1, 2, 3. DOI: 10.1107/S2056989023002517/vm22761sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989023002517/vm22762sup3.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S2056989023002517/vm22763sup4.hklStructure factors: contains datablock(s) 3. DOI: Click here for additional data file.10.1107/S2056989023002517/vm22761sup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023002517/vm22762sup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023002517/vm22763sup7.cmlSupporting information file. DOI: 2248697, 2248696, 2248695CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Blood Cancer Journal (2023) 13:111 10.1038/s41408-023-00883-x, published online 20 July 2023Correction to: N\u2009=\u20093111)\u201d and \u201cNon-ASCT (N\u2009=\u20092437)\u201d were assigned in incorrect lines. Correct Table\u00a0In Table\u00a0ASCT N\u2009=\u2009437\u201d wereThe original article has been corrected."} +{"text": "In the published article, there was an error in the name and abbreviation of the measure \u201cmean corpuscular hemoglobin concentration\u201d abbreviated to \u201cMCHC\u201d. It should be \u201cmean corpuscular hemoglobin\u201d and abbreviated to \u201cMCH\u201d. Throughout the text of the article, \u201cmean corpuscular hemoglobin concentration\u201d has been corrected to \u201cmean corpuscular hemoglobin\u201d and the abbreviation \u201cMCHC\u201d has been corrected to \u201cMCH\u201d.In the published article, there was an error in the header, title and legend for The corrected In the published article, there was an error in the legend for In the published article, there was an error. A correction has been made to the section Results, subsection \u201cAnalysis of anemia prevalence and hematological phenotype indexes in pregnant women with various thalassemia genotypes\u201d, paragraph 1. This sentence previously stated:SEA, \u2013\u03b13.7/\u2013\u03b13.7, \u2013\u03b13.7/\u2013\u03b14.2, and \u2013\u03b14.2/\u2013\u03b14.2 ranged from 35.96 to 37.82% (P < 0.05).\u201d\u201cThe anemia rates of \u03b1\u03b1/\u2013The corrected sentence appears below:SEA, \u2013\u03b13.7/\u2013\u03b13.7, \u2013\u03b13.7/\u2013\u03b14.2, and \u2013\u03b14.2/\u2013\u03b14.2 ranged from 35.96 to 37.82% (P > 0.05).\u201d\u201cThe anemia rates of \u03b1\u03b1/\u2013A correction has been made to the section Discussion, subsection \u201cAnalysis of anemia prevalence and hematological phenotype indexes of pregnant women with various thalassemia genotypes\u201d, paragraph 1. This sentence previously stated:SEA, \u2013\u03b13.7/\u2013\u03b13.7, \u2013\u03b14.2/\u2013\u03b14.2, and \u2013\u03b13.7/\u2013\u03b14.2 ranged from 35.96 to 37.82% (P < 0.05), and no significant differences were observed in Hb levels among those genotypes.\u201d\u201cThe anemia rate among carriers of \u03b1\u03b1/\u2013The corrected sentence appears below:SEA, \u2013\u03b13.7/\u2013\u03b13.7, \u2013\u03b14.2/\u2013\u03b14.2, and \u2013\u03b13.7/\u2013\u03b14.2 ranged from 35.96 to 37.82% (P > 0.05), and no significant differences were observed in Hb levels among those genotypes.\u201d\u201cThe anemia rate among carriers of \u03b1\u03b1/\u2013A correction has been made to the section Discussion, subsection \u201cPrevalence of iron deficiency in anemic pregnant women with various genotypes of thalassemia\u201d, paragraph 1. This sentence previously stated:\u201cAlthough there is no large sample, multicenter randomized controlled study that has yet confirmed the need for routine iron supplementation in PW with thalassemia minor genotypes, we suggest that the iron load of APW with \u03b2- thalassemia minor genotypes should be monitored dynamically during iron supplementation\u201dThe corrected sentence appears below:\u201cAlthough there is no large sample, multicenter randomized controlled study that has yet confirmed the need for routine iron supplementation in PW with thalassemia minor genotypes, we suggest that the ferritin levels of APW with \u03b2- thalassemia minor genotypes should be monitored dynamically during iron supplementation\u201dThe authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Wu et al. show that the mitotic kinesin CENP-E promotes full expansion of the outer layer of the kinetochore known as the fibrous corona. This new function of CENP-E is independent of its motor activity. The findings expand the roles of CENP-E in spindle assembly and chromosome congression in mitosis. Correct chromosome segregation during cell division depends on proper connections between spindle microtubules and kinetochores. During prometaphase, kinetochores are temporarily covered with a dense protein meshwork known as the fibrous corona. Formed by oligomerization of ROD/ZW10/ZWILCH-SPINDLY (RZZ-S) complexes, the fibrous corona promotes spindle assembly, chromosome orientation, and spindle checkpoint signaling. The molecular requirements for formation of the fibrous corona are not fully understood. Here, we show that the fibrous corona depends on the mitotic kinesin CENP-E and that poorly expanded fibrous coronas after CENP-E depletion are functionally compromised. This previously unrecognized role for CENP-E does not require its motor activity but instead is driven by farnesyl modification of its C-terminal kinetochore- and microtubule-binding domain. We show that in cells, CENP-E binds Spindly and recruits RZZ-S complexes to ectopic locations in a farnesyl-dependent manner. CENP-E is recruited to kinetochores following RZZ-S, and\u2014while not required for RZZ-S oligomerization per se\u2014promotes subsequent fibrous corona expansion. Our comparative genomics analyses suggest that the farnesylation motif in CENP-E orthologs emerged alongside the full RZZ-S module in an ancestral lineage close to the fungi\u2013animal split (Obazoa), revealing potential conservation of the mechanisms for fibrous corona formation. Our results show that proper spindle assembly has a potentially conserved non-motor contribution from the kinesin CENP-E through stabilization of the fibrous corona meshwork during its formation. The cell division cycle culminates in the segregation of duplicated sister chromatids to nascent daughter cells. Segregation is driven by the microtubule-based spindle apparatus . MicrotuThe fibrous corona houses over two dozen proteins to perform its various roles in spindle assembly and chromosome segregation . It is fcomet, the Cyclin B1\u2013CDK1 kinase complex, the CLASP1/2 microtubule dynamics regulators, and the microtubule-binding protein centromere protein F was fully sufficient to restore fibrous corona formation after CENP-E loss, we next attempted to further narrow down the essential region. Expression in HeLa cells of a series of MT-binding domain mutants of CENP-E 2111-C revealedC2698A delayed chromosome congression have sm ZWINT-1 . The kin ZWINT-1 . In cont ZWINT-1 . Further ZWINT-1 , showing ZWINT-1 or expre ZWINT-1 abolisheOur data thus far showed that farnesylated CENP-E is important for fibrous corona formation and that CENP-E can quite robustly recruit RZZ-S to ectopic locations in a manner dependent on its farnesylation. To understand more about how CENP-E promotes fibrous corona formation, we asked when and how CENP-E localizes to kinetochores, relative to RZZ-S. Again we made use of the monopolar spindle assay, in which fibrous corona proteins are depleted from kinetochores and accumulate at the spindle pole upon chemical inhibition of CENP-E. Subsequent addition of nocodazole to these cells to deplete microtubules caused detectable relocalization of ZW10 back to the kinetochores within \u223c1 min, followed by CENP-E after \u223c8 min . Importa\u0394N) bypasses the need for regulatory mechanisms of fibrous corona formation and does not bind dynein/dynactin, resulting in persistent fibrous coronas should be able to bypass this step in the absence of CENP-E. This is not so, showing CENP-E contributes to a parallel or downstream event. An alternative hypothesis is that CENP-E is an integral component of the RZZ-S meshwork. However, RZZ-S can oligomerize in vitro without CENP-E . Our current analysis pinpoints emergence of SPINDLY one branch earlier than the cysteine in CENP-E, but we note that the CENP-E ortholog sequences in Breviata are incomplete. Better sampling of genomes from Breviata species might therefore reveal FL CENP-E ORF sequences with a cysteine at \u22124 from the carboxy terminus, which would indicate near-simultaneous emergence of SPINDLY and the potential for CENP-E farnesylation. Beyond SPINDLY, the RZZ-S complex and the potential for CENP-E farnesylation show strongly cohesive presence/absence in Obazoa, with notable coherent losses close in the species tree. Interestingly, kinetochores in colchicine-treated Drosophila S2 cells do not appear to form clear fibrous coronas supplemented with 9% tetracycline-free fetal bovine serum and 100 \u03bcg/ml penicillin-streptomycin .\u0394N, and LAP-SPINDLY\u0394N/C602A in pCDNA5 vector were previously cloned by PCR-based strategy; SPINDLY\u0394N: SPINDLY 65-605; SPINDLY\u0394N/C602A: SPINDLY 65-605 with C602A to the C-terminal CENP-E 2111-C (comprising KT-binding domain and MT-binding domain). LAP-tagged CENP-E 2,111\u20132,697^CAAXLAP-ROD in pCDNA5 vector was cloned based on the ROD cDNA, which was a kind gift from Reto Gassmann , TagRFP-CENP-E 2111-C and H2B-mScarlet were cloned into the pLVX-IRES-Puro vector (Clontech) by PCR-based Gibson assembly method.CENP-E-1aa-XhoI-GA-F: 5\u2032-CGG\u200bAGG\u200bTGG\u200bATC\u200bCAC\u200bTAG\u200bTCT\u200bCGA\u200bGAT\u200bGGC\u200bGGA\u200bGGA\u200bAGG\u200bAGC\u200bCGT-3\u2032CENP-E-2,701aa-NotI-GA-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTT\u200bTTG\u200bCAC\u200bTCA\u200bGGC\u200bACA\u200bTC-3\u2032CENP-E-1,301-BamHI-F: 5\u2032-GCA\u200bCAT\u200bGGA\u200bCAG\u200bCGG\u200bAGG\u200bTGG\u200bATC\u200bCAC\u200bTCA\u200bGGA\u200bAAC\u200bAAT\u200bGAA\u200bTGA\u200bACT\u200bGGA\u200bGT-3\u2032CENP-E-2,111-BamHI-F: 5\u2032-GCA\u200bCAT\u200bGGA\u200bCAG\u200bCGG\u200bAGG\u200bTGG\u200bATC\u200bCGA\u200bTGA\u200bTCA\u200bTTA\u200bTGA\u200bGTG\u200bCTT\u200bGAA\u200bTAG\u200bATT\u200bGTC\u200bTCT-3\u2032CENP-E 2,599-NotI-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAT\u200bTGT\u200bTTG\u200bTGA\u200bGCC\u200bTCT\u200bCTT\u200bTTA\u200bAGG\u200bGTT-3\u2032CENP-E 2,643-NotI-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAA\u200bCAA\u200bGAT\u200bTTT\u200bGGT\u200bGAT\u200bTCC\u200bTTT\u200bGGC\u200bACA-3\u2032CENP-E-2,600-mutation10AA-GA-R: 5\u2032-TGC\u200bAGC\u200bCGC\u200bTGC\u200bAGC\u200bTGC\u200bCGC\u200bTGC\u200bGGC\u200bAGC\u200bTTG\u200bTTT\u200bGTG\u200bAGC\u200bCTC\u200bTCT\u200bTTT\u200bAAG\u200bGGT\u200bT-3\u2032CENP-E-2,609-mutation10AA-GA-F: 5\u2032-GCG\u200bGCA\u200bGCT\u200bGCA\u200bGCG\u200bGCT\u200bGCA\u200bAAA\u200bGTG\u200bACT\u200bGGA\u200bACA\u200bGCT\u200bTCT\u200bAAA\u200bAAG\u200bAAA\u200bC-3\u2032CENP-E-2,610-mutation10AA-GA-R: 5\u2032-TGC\u200bAGC\u200bCGC\u200bTGC\u200bAGC\u200bTGC\u200bCGC\u200bTGC\u200bGGC\u200bAGC\u200bAGG\u200bAGA\u200bCTT\u200bTGG\u200bAGA\u200bATT\u200bCTC\u200bACA\u200bAG-3\u2032CENP-E-2,619-mutation10AA-GA-F: 5\u2032-GCG\u200bGCA\u200bGCT\u200bGCA\u200bGCG\u200bGCT\u200bGCA\u200bCAA\u200bATT\u200bACA\u200bCCC\u200bTCT\u200bCAA\u200bTGC\u200bAAG\u200bGAA\u200bC-3\u2032CENP-E-2,620-mutation10AA-GA-R: 5\u2032-TGC\u200bAGC\u200bCGC\u200bTGC\u200bAGC\u200bTGC\u200bCGC\u200bTGC\u200bGGC\u200bAGC\u200bTTT\u200bCTT\u200bTTT\u200bAGA\u200bAGC\u200bTGT\u200bTCC\u200bAGT\u200bCAC-3\u2032CENP-E-2,629-mutation10AA-GA-F: 5\u2032-GCG\u200bGCA\u200bGCT\u200bGCA\u200bGCG\u200bGCT\u200bGCA\u200bAAT\u200bTTA\u200bCAA\u200bGAT\u200bCCT\u200bGTG\u200bCCA\u200bAAG\u200bGAA\u200bTC-3\u2032CENP-E-2,630-mutation10AA-GA-R: 5\u2032-TGC\u200bAGC\u200bCGC\u200bTGC\u200bAGC\u200bTGC\u200bCGC\u200bTGC\u200bGGC\u200bAGC\u200bCCG\u200bTTC\u200bCTT\u200bGCA\u200bTTG\u200bAGA\u200bGGG\u200bTG-3\u2032CENP-E-2,639-mutation10AA-GA-F: 5\u2032-GCG\u200bGCA\u200bGCT\u200bGCA\u200bGCG\u200bGCT\u200bGCA\u200bCCA\u200bAAA\u200bTCT\u200bTGT\u200bTTT\u200bTTT\u200bGAT\u200bAGC\u200bCGA\u200bTC-3\u2032CENP-E-2,640-mutation10AA-GA-R: 5\u2032-TGC\u200bAGC\u200bCGC\u200bTGC\u200bAGC\u200bTGC\u200bCGC\u200bTGC\u200bGGC\u200bAGC\u200bTGA\u200bTTC\u200bCTT\u200bTGG\u200bCAC\u200bAGG\u200bATC\u200bTTG-3\u2032CENP-E-2,649-mutation10AA-GA-F: 5\u2032-GCG\u200bGCA\u200bGCT\u200bGCA\u200bGCG\u200bGCT\u200bGCA\u200bAAG\u200bTCT\u200bTTA\u200bCCA\u200bTCA\u200bCCT\u200bCAT\u200bCCA\u200bG-3\u2032CENP-E-2,650-mutation10AA-GA-R: 5\u2032-TGC\u200bAGC\u200bCGC\u200bTGC\u200bAGC\u200bTGC\u200bCGC\u200bTGC\u200bGGC\u200bAGC\u200bTGA\u200bTCG\u200bGCT\u200bATC\u200bAAA\u200bAAA\u200bACA\u200bAGA\u200bTTT\u200bTG-3\u2032CENP-E-2,659-mutation10AA-GA-F: 5\u2032-GCG\u200bGCA\u200bGCT\u200bGCA\u200bGCG\u200bGCT\u200bGCA\u200bTAT\u200bTTT\u200bGAT\u200bAAC\u200bTCA\u200bAGT\u200bTTA\u200bGGC\u200bCTT\u200bTGT\u200bC-3\u2032CENP-E-2,660-mutation10AA-GA-R: 5\u2032-TGC\u200bAGC\u200bCGC\u200bTGC\u200bAGC\u200bTGC\u200bCGC\u200bTGC\u200bGGC\u200bAGC\u200bGCG\u200bAAC\u200bTGG\u200bATG\u200bAGG\u200bTGA\u200bTGG\u200bTAA-3\u2032CENP-E-2,669-mutation10AA-GA-F: 5\u2032-GCG\u200bGCA\u200bGCT\u200bGCA\u200bGCG\u200bGCT\u200bGCA\u200bCCA\u200bGAG\u200bGTG\u200bCAA\u200bAAT\u200bGCA\u200bGGA\u200bG-3\u2032CENP-E-2,670-mutation10AA-GA-R: 5\u2032-TGC\u200bAGC\u200bCGC\u200bTGC\u200bAGC\u200bTGC\u200bCGC\u200bTGC\u200bGGC\u200bAGC\u200bACA\u200bAAG\u200bGCC\u200bTAA\u200bACT\u200bTGA\u200bGTT\u200bATC\u200bAAA\u200bAT-3\u2032CENP-E-2,679-mutation10AA-GA-F: 5\u2032-GCG\u200bGCA\u200bGCT\u200bGCA\u200bGCG\u200bGCT\u200bGCA\u200bGTG\u200bGAT\u200bTCT\u200bCAG\u200bCCA\u200bGGT\u200bCCT\u200bTG-3\u2032CENP-E-2,680-mutation10AA-GA-R: 5\u2032-TGC\u200bAGC\u200bCGC\u200bTGC\u200bAGC\u200bTGC\u200bCGC\u200bTGC\u200bGGC\u200bAGC\u200bACT\u200bCTC\u200bTGC\u200bTCC\u200bTGC\u200bATT\u200bTTG\u200bC-3\u2032CENP-E-2,689-mutation10AA-GA-F: 5\u2032-GCG\u200bGCA\u200bGCT\u200bGCA\u200bGCG\u200bGCT\u200bGCA\u200bTCC\u200bTCA\u200bGGC\u200bAAG\u200bGAT\u200bGTG\u200bCCT\u200bG-3\u2032CENP-E-2,689-NotI-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAG\u200bGCG\u200bTGC\u200bCAA\u200bGGA\u200bCCT\u200bGGC\u200bTGA\u200bG-3\u2032CENP-E-2,701-NotI-S2690A-R2: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTT\u200bTTG\u200bCAC\u200bTCA\u200bGGC\u200bACA\u200bTCC\u200bTTG\u200bCCT\u200bGAA\u200bGCG\u200bGCG\u200bTGC\u200bCAA\u200bGGA\u200bCCT\u200bG-3\u2032CENP-E-2,701-NotI-S2691A-R2: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTT\u200bTTG\u200bCAC\u200bTCA\u200bGGC\u200bACA\u200bTCC\u200bTTG\u200bCCA\u200bGCG\u200bGAG\u200bGCG\u200bTGC\u200bCAA\u200bGGA\u200bC-3\u2032CENP-E-2,701-NotI-K2693A-R2: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTT\u200bTTG\u200bCAC\u200bTCA\u200bGGC\u200bACA\u200bTCA\u200bGCG\u200bCCT\u200bGAG\u200bGAG\u200bGCG\u200bTGC\u200bCAA\u200bGGA\u200bC-3\u2032CENP-E-2,701-NotI-D2694A-R2: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTT\u200bTTG\u200bCAC\u200bTCA\u200bGGC\u200bACA\u200bGCC\u200bTTG\u200bCCT\u200bGAG\u200bGAG\u200bGCG\u200bTGC\u200bCAA\u200bGGA\u200bC-3\u2032CENP-E-2,701-NotI-V2695A-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTT\u200bTTG\u200bCAC\u200bTCA\u200bGGA\u200bGCA\u200bTCC\u200bTTG\u200bCCT\u200bGAG\u200bGAG\u200bGCG\u200bTG-3\u2032CENP-E-2,701-NotI-P2696A-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTT\u200bTTG\u200bCAC\u200bTCA\u200bGCC\u200bACA\u200bTCC\u200bTTG\u200bCCT\u200bGAG\u200bGAG\u200bGC-3\u2032CENP-E-2,701-NotI-E2697A-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTT\u200bTTG\u200bCAA\u200bGCA\u200bGGC\u200bACA\u200bTCC\u200bTTG\u200bCCT\u200bGAG\u200bGAG-3\u2032CENP-E-C-C2698A-R2: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTT\u200bTTA\u200bGCC\u200bTCA\u200bGGC\u200bACA\u200bTCC\u200bTTG\u200bCCT\u200bGAG\u200bGAG\u200bGC-3\u2032CENP-E-2,701-NotI-K2699A-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTA\u200bGCG\u200bCAC\u200bTCA\u200bGGC\u200bACA\u200bTCC\u200bTTG\u200bCC-3\u2032CENP-E-2,701-NotI-T2700A-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGCT\u200bTTG\u200bCAC\u200bTCA\u200bGGC\u200bACA\u200bTCC\u200bTTG\u200bC-3\u2032CENP-E-2,701-NotI-Q2701A-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAA\u200bGCA\u200bGTT\u200bTTG\u200bCAC\u200bTCA\u200bGGC\u200bACA\u200bTCC\u200bT-3\u2032CENP-E-2,701-NotI-C2698S-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bTGA\u200bGTT\u200bTTT\u200bGAC\u200bTCA\u200bGGC\u200bACA\u200bTCC\u200bTTG\u200bCCT\u200bGAG\u200bGAG\u200bGCG\u200bTG-3\u2032CENP-E^KRAS_caax_R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCC\u200bTAC\u200bATT\u200bATA\u200bATG\u200bCAC\u200bTCA\u200bGGC\u200bACA\u200bTCC\u200bTTG\u200bCCT\u200bGAG\u200bGAG\u200bGCG\u200bTG-3\u2032TagRFP-kozak-EcoR1-F: 5\u2032-CTA\u200bCTA\u200bGAG\u200bGAT\u200bCTA\u200bTTT\u200bCCG\u200bGTG\u200bAAT\u200bTCG\u200bCCA\u200bCCA\u200bTGG\u200bTGT\u200bCTA\u200bAGG\u200bGCG\u200bAAG\u200bAGC\u200bTGA\u200bTTA\u200bAG-3\u2032TagRFP-C-toCENP-E2111-R: 5\u2032-TCT\u200bATT\u200bCAA\u200bGCA\u200bCTC\u200bATA\u200bATG\u200bATC\u200bATC\u200bGGA\u200bTCC\u200bACC\u200bTCC\u200bGCT\u200bATT\u200bAAG\u200bTTT\u200bGTG\u200bCCC\u200bCAG\u200bTTT\u200bGCT-3\u2032ROD-1aa-XhoI-GA-F: 5\u2032-CGG\u200bAGG\u200bTGG\u200bATC\u200bCAC\u200bTAG\u200bTCT\u200bCGA\u200bGAT\u200bGTG\u200bGAA\u200bTGA\u200bTAT\u200bTGA\u200bGCT\u200bGCT\u200bAAC\u200bAAA\u200bTG-3\u2032ROD-1,100aa-overlap-GA-R: 5\u2032-AAT\u200bAGC\u200bAAT\u200bTTC\u200bCCA\u200bGTG\u200bTCA\u200bGCA\u200bTT-3\u2032ROD-1,100aa-overlap-GA-F: 5\u2032-AAT\u200bGCT\u200bGAC\u200bACT\u200bGGG\u200bAAA\u200bTTG\u200bCTA\u200bTT-3\u2032ROD-2,209aa-NotI-GA-R: 5\u2032-GCG\u200bGGT\u200bTTA\u200bAAC\u200bGGG\u200bCCC\u200bGCG\u200bGCC\u200bGCT\u200bTAC\u200bGAT\u200bAAT\u200bCCA\u200bCTA\u200bAGA\u200bAAC\u200bATC\u200bTTC\u200bAGA\u200bAT-3\u2032TurboID-N-BamHI-F: 5\u2032-GCA\u200bCAT\u200bGGA\u200bCAG\u200bCGG\u200bAGG\u200bTGG\u200bATC\u200bCAA\u200bAGA\u200bCAA\u200bTAC\u200bTGT\u200bGCC\u200bTCT\u200bGAA\u200bGCT-3\u2032TurboID-C-BamHI-R: 5\u2032-TCA\u200bAGC\u200bACT\u200bCAT\u200bAAT\u200bGAT\u200bCAT\u200bCGG\u200bATC\u200bCGC\u200bTGA\u200bATT\u200bCCT\u200bTTT\u200bCGG\u200bCAG\u200bACC\u200bGCA\u200bGAC\u200bT-3\u2032CENP-E-1aa-BamHI-cut-GA-F: 5\u2032-GCA\u200bCAT\u200bGGA\u200bCAG\u200bCGG\u200bAGG\u200bTGG\u200bATC\u200bCAT\u200bGGC\u200bGGA\u200bGGA\u200bAGG\u200bAGC\u200bCGT\u200bG-3\u2032CENP-E-426aa-BamHI-cut-GA-R: 5\u2032-ATT\u200bCAA\u200bGCA\u200bCTC\u200bATA\u200bATG\u200bATC\u200bATC\u200bGGA\u200bTCC\u200bGCC\u200bAAG\u200bGCA\u200bCCA\u200bAGT\u200bAAC\u200bTCT\u200bTC-3\u2032Lentiviruses were produced by cotransfection of HEK 293T cells with the lentiviral vector containing TagRFP-CENP-E 2111-C or H2B-mScarlet and separate plasmids that express Tat, vesicular stomatitis virus G glycoprotein, Rev, and Gag-Pol, with the transfection reagent of Fugene HD . 72 h after transfection, cell culture supernatant was harvested and filtered.RPE1 FlpIn cell line stably expressing EB3-TagRFPT-ires-GFP-ZWILCH was made previously by transduction of viruses that were produced by cotransfection of HEK 293T cells with the lentiviral vector containing gene of EB3-TagRFPT-ires-GFP-Zwilch, which was cloned into the pLVX-IRES-Puro vector .\u0394N were made previously by cotransfection of pCDNA5-constructs with gene of interest and construct of pOG44 recombinase in a 1:2 ratio with electroporation machine (Amaxa Nucleofector II) using protocol U-017. 3 d after transfection, RPE1 cells were selected with 100 \u03bcM hygromycin for about 3 wk (RPE1 FlpIn cell line inducible expressing LAP-SPINDLY FL and LAP- SPINDLYout 3 wk .RPE1 FlpIn cell lines were cotransfected with pCDNA5-constructs with gene of interest and construct of pOG44 recombinase in a 1:2 ratio with an electroporation machine (Amaxa Nucleofector II) using protocol U-017. 3 d after transfection, RPE1 cells were selected with 100 \u03bcM hygromycin for about 3 wk.HeLa FlpIn were cotransfected with pCDNA5 constructs with the gene of interest and construct of pOG44 recombinase in a 1:9 ratio with Fugene HD under the manufacturer\u2019s instructions. 1 d after transfection, HeLa cells were selected with 200 \u03bcM hygromycin for about 3 wk.To generate HeLa FlpIn cells stably expressing TagRFP-CENP-E 2111-C and inducible expressing LAP-ROD, HeLa FlpIn cells inducible expressing LAP-ROD were infected with lentiviruses and selected with 1 \u03bcg/ml puromycin for 1 wk.To generate HeLa FlpIn cell stably expressing H2B-mScarlet, HeLa FlpIn were infected with lentiviruses and selected with 1 \u03bcg/ml puromycin for 1 wk.To generate HeLa FlpIn cells inducible expressing CENP-E FL WT or CENP-E FL C2698A and stably expressing H2B-mScarlet, HeLa FlpIn cells inducible expressing CENP-E FL WT or CENP-E FL C2698A were infected with lentiviruses and selected with 1 \u03bcg/ml puromycin for 1 wk.The siRNAs targeting GAPDH , CENP-E for 15 h (overnight) for fibrous corona formation before the fixation. HeLa FlpIn cells were treated with 6.6 \u03bcM nocodazole for 6\u223c8 h before the fixation. For cells transfected with siRNA, nocodazole was added 48 h after siRNA transfection. For the rescue experiments, 1 \u03bcg/ml doxycycline was added to cells 40 h after siRNA transfection to induce the expression of protein mutants. For nocodazole washout assay, 6.6 \u03bcM nocodazole was added to the culture medium for 15 h before washout. For CENP-E inhibition, cells were treated with 250 nM GSK923295 for 15 h (overnight). For farnesyl transferase inhibition, cells were treated with 5 \u00b5m lonafarnib for 15 h (overnight). For the fast disassembly of microtubules, cells were treated with 13.2 \u03bcM nocodazole for 4 h. Cells were treated with 10 mM STLC for 15 h (overnight). For the experiments in which translocated fibrous corona proteins were released from spindle poles, cells were cotreated with STLC and GSK923295 for 15 h (overnight) and were further treated with 13.2 \u03bcM nocodazole for indicated time before fixation. In the TurboID pulldown experiment, HeLa cells were treated with 1 \u03bcg/ml doxycycline for 24 and 1 h of 250 \u03bcM biotin before harvest. For spindle end-on attachment experiments, HeLa cells were cotreated with STLC and ZM447439 for 6 h before fixation.Glued . We used the following secondary antibodies: Goat anti-guinea pig Alexa Fluor 647 , Goat anti-rabbit Alexa Fluor 488 , Goat anti-rabbit Alexa Fluor 568 , Goat anti-rabbit Alexa Fluor 405 , Goat anti-mouse Alexa Fluor 568 , Goat anti-mouse Alexa Fluor 647 , and GFP-Booster Atto 488 .We used guinea pig polyclonal antibody against CENPC , and rabbit polyclonal antibodies against CENP-E , ZW10 , and were blocked with 2% BSA diluted in PBST for 30 min. After blocking, cells were incubated with primary antibodies diluted in PBST containing 2% BSA for 1 h at room temperature in humid conditions. Subsequently, cells were washed three times with PBST and incubated with secondary antibodies diluted in PBST together with or without DAPI for 1 h at room temperature. Then, cells were washed three times with PBST. In the end, coverslips were sequentially rinsed in 70% and 100% ethanol, air-dried, and mounted on glass slides with Prolong Gold antifade.n = 1.5160 using SoftWorx 6.0 software . Images were acquired as z-stacks at 0.2-\u03bcm intervals for 32 stacks using Photometrics Coolsnap HQ2 cameral (Roper Scientific) and deconvolved using SoftWoRx. The images are maximum-intensity projections of deconvoluted stacks.All images were acquired on a deconvolution system with a 100\u00d7/1.40 NA UPlanSApo objective (Olympus) and imaging medium with https://fiji.sc). For quantification of protein levels, all images of immunostaining experiments were acquired with identical illumination settings. Protein levels near the kinetochores were determined on maximum projections of z-stacks images using an ImageJ macro . 488 and 561 nm lasers were used for sample excitation and images were acquired using an Andor iXon-888 EMCCD camera. Nocodazole washout was done during the imaging with a warm culture medium. Before and after nocodazole washout, cells were kept at 37\u00b0C and 5% CO2 using a cage incubator and Boldline temperature/CO2 controller (OKO-Lab).For imaging of RPE1 cells coexpressing EB3-TagRFPT and GFP-Zwilch in nocodazole washout assay, RPE1 cells were cultured in a 24-well glass bottom plate . Live-cell imaging was performed on a Nikon TiE microscope, controlled by NIS Elements software , by acquiring images every 20 or 30 s with z-stacks at 1-\u03bcm intervals for 15 stacks at 1 \u2009\u00d7 \u20091 binning on an Andor CSU-W1 spinning disk (50 \u03bcm disk) with Nikon 100\u00d7 1.45 NA oil objective and immersion oil with n = 1.406 . 488 and 561 nm lasers were used for sample excitation and images were acquired using an Andor iXon-888 EMCCD camera. Cells were kept at 37\u00b0C and 5% CO2 using a cage incubator and Boldline temperature/CO2 controller (OKO-Lab).Imaging of HeLa FlpIn cells expressing H2B-mScarlet and HeLa FlpIn cells coexpressing LAP-CENP-E FL WT or LAP-CENP-E FL C2698A with H2B-mScarlet that were cultured in 8-well plates was performed on a Nikon TiE microscope, controlled by NIS Elements software , by acquiring images every 3 min with z-stacks at 2-\u03bcm intervals for eight stacks at 1\u2009 \u00d7 \u20091 binning on an Andor CSU-W1 spinning disk (50-\u03bcm disk) with Nikon 30\u00d7 silicon objective and silicon immersion oil with In the TurboID-based proteins pulldown experiments, HeLa cells inducible expressing LAP-TurboID-CENP-E 2111-C mutants were treated with 1 \u03bcg/ml doxycycline for 24 h to induce the expression of LAP-TurboID-CENP-E 2111-C mutants and 1 h of 250 \u03bcM biotin before harvest. The cells were harvested by trypsinization and centrifuge. The cells were washed once with ice-cold PBS and lysated with lysis buffer on ice for 10 min. After lysis, the samples were centrifuged at 14,000 rpm for 20 min at 4\u00b0C. The supernatants of the centrifuged samples were incubated with the streptavidin magnetic beads for 1 h in a 4\u00b0C cold room. The beads were washed with wash buffer three times. The wash buffer was removed after the final wash and the beads were stored in a \u221280\u00b0C freezer.2 Fold Change and \u2212log10 P value higher than 2. The program used for the analyses was R (version 4.0.4) through RStudio (version 1.5.64).Precipitated proteins were denatured and alkylated in 50 \u00b5l 8 M urea, 1 M ammonium bicarbonate (ABC) containing 10 mM tris (2-carboxyethyl) phosphine hydrochloride and 40 mM 2-chloro-acetamide for 30 min. After fourfold dilution with 1 M ABC and digestion with trypsin (20 \u00b5g/200 \u00b5l), peptides were separated from the beads and desalted with homemade C-18 stage tips (3 M), eluted with 80% acetonitrile (ACN) and, after evaporation of the solvent in the speedvac, redissolved in buffer A (0.1% formic acid [FA]). After separation on a 30-cm pico-tip column in-house packed with C-18 material using a 140 min gradient , delivered by an easy-nLC 1,000 (Thermo Fisher Scientific), peptides were electrosprayed directly into a Orbitrap Fusion Tribrid Mass Spectrometer (Thermo Fisher Scientific). The latter was set in data-dependent top speed mode with a cycle time of 1 s, in which the full scan over the 400\u20131,500 mass range was performed at a resolution of 240,000. Most intense ions were isolated by the quadrupole, where after they were fragmented with a higher-energy collisional dissociation collision energy of 30%. The maximum injection time of the ion trap was set to 50 ms with injection of ions for all available parallelizable time. Raw data were analyzed with MaxQuant (version 1.6.3.4) using the Homo Sapiens (taxonomy ID: 9606) fasta file, extracted from UniprotKB. To determine proteins of interest, the protein group output file was used to perform a differential enrichment analysis. Proteins with less than one unique peptide and proteins that have not been identified in at least two out of three of the replicates of one condition were filtered out. Then, a background correction and normalization of the data were performed by variance stabilizing transformation, shifting and scaling the protein intensities by sample group. A left-shifted Gaussian distribution randomization was used to impute since the data presented a pattern of missingness not at random. Finally, a differential enrichment analysis was performed to identify those proteins that were differentially enriched and selected those falling inside the threshold for logThe mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner Preprint), with some minor adjustments detailed in The eukaryotic sequence database used for the evolutionary analysis consisted of 177 predicted proteomes derived from genomes or transcriptomes and spans a representative set of species across the eukaryotic tree of life. This dataset is derived from earlier work and Prism 9 . Data distribution was assumed to be normal but this was not formally tested. The results of The data generated are available in the published article and its online supplemental material. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD040338."} +{"text": "In the title compound, which is related to the herbicide flufenacet, the amide group and fluoro\u00adbenzene ring are almost perpendicular. A short O\u22ef\u03c0 contact is observed in the crystal. 12H16FNO3S, which is related to the herbicide flufenacet, are presented. The dihedral angle between the amide group and the fluorinated benzene ring is 87.30\u2005(5)\u00b0 and the N\u2014C\u2014C\u2014S torsion angle defining the orientation of the methyl\u00adsulfonyl substituent relative to the amide group is 106.91\u2005(11)\u00b0. In the crystal, inversion-related mol\u00adecules form dimers as a result of pairwise C\u2014H\u22efO hydrogen bonds, which appear to be reinforced by short O\u22ef\u03c0 contacts [O\u22efCg = 3.0643\u2005(11)\u2005\u00c5]. A Hirshfeld surface analysis was used to qu\u00adantify the various types of inter\u00admolecular contacts, which are dominated by H atoms.The synthesis and crystal structure of the title compound, C For example, substituted phenyl\u00adacetamides and their use as protease inhibitors was reported by Kreutter et al. -yl]acetamides was given by Ahmad et al. (2013N4-substituted sulfonamide\u2013acetamide derivatives as di\u00adhydro\u00adfolate reductase (DHFR) inhibitors was reported by Hussein et al. (2019N-(substituted phen\u00adyl)-N-(substituted)acetamide derivatives as potent analgesic agents was described by Verma et al. -2-phenyl\u00adacetamide derivatives as potential anti\u00admycobacterial agents was reported by G\u00fczel-Akdemir et al. , systematic name N-(4-fluoro\u00adphen\u00adyl)-N-propan-2-yl-2-{\u00adoxy}acetamide, is an herbi\u00adcide, xenobiotic and environmental contaminant Fig.\u00a01.2.I, the nitro\u00adgen atom of the amide group is close to planar, the sum of bond angles about N1 being 358.92\u2005(19)\u00b0, which places N1 0.0862\u2005(14)\u2005\u00c5 from the plane passing through C1, C4, and C7. The amide group is also almost planar, having an r.m.s. deviation from the mean plane of N1, C1, O1, C2 of 0.0095\u2005\u00c5 [maximum = 0.0165\u2005(11)\u2005\u00c5 for C1], and is almost perpendicular to the fluoro\u00adbenzene ring (C7\u2013C12), subtending a dihedral angle of 87.30\u2005(5)\u00b0. The overall conformation of the mol\u00adecule is defined by the torsion angles C7\u2014N1\u2014C1\u2014C2 [14.68\u2005(17)\u00b0], N1\u2014C1\u2014C2\u2014S1 [106.91\u2005(11)\u00b0], C1\u2014C2\u2014S1\u2014C3 [74.53\u2005(10)\u00b0] and by the orientation of the ipropyl group, e.g., C1\u2014N1\u2014C4\u2014C6 [139.85\u2005(13)\u00b0]. Otherwise, all bond lengths and angles lie within the expected ranges.In the crystal structure of 3.I 1 \u2013 x, 1 \u2013 y, 1 \u2013 z]. These dimers also feature close contacts between the sulfone O3 atom and the inversion-related benzene ring to give an O3\u22efCg(C7\u2013C12)i distance of 3.0643\u2005(11)\u2005\u00c5 . The other weak C\u2014H\u22efO inter\u00adactions involve inversion, translation, and c-glide related mol\u00adecules (Table\u00a01a). A Hirshfeld surface analysis using CrystalExplorer .There are no strong hydrogen bonds in the crystal structure of I Fig.\u00a02, but thes Table\u00a01a. A Hir4.et al., 2016N-phenyl\u00adacetamide with \u2018any non-H group\u2019 attached at the nitro\u00adgen atom, the 4-position of the benzene ring, and replacing one hydrogen of the methyl group, yielded 259 hits. A similar fragment, but with \u2018any halogen\u2019 at the 4-position on the ring, gave 92 hits. With the halogen restricted to fluorine, twelve hits were returned, and with an isopropyl group attached to the nitro\u00adgen atom, only one match was found: CSD refcode QEMHOG was presented by Gung et al. (2008A search of the CSD for non-bonded close contacts (up to 3.1\u00c5) between S=O oxygen atoms and a benzene-ring centroid (with \u2018any substituent\u2019) returned 154 hits, none of which have much else in common with al. 2008.d-threo-2,2-di\u00adchloro-N-{2-hy\u00addroxy-1-(hy\u00addroxy\u00admeth\u00adyl)-2-[4-(methyl\u00adsulfon\u00adyl)phen\u00adyl]eth\u00adyl}acet\u00ada\u00ad\u00admide -2-hy\u00addroxy-2-[4- (methyl\u00adsulfon\u00adyl)phen\u00adyl]eth\u00adyl}acetamide -2-(2-{3-[4-(methyl\u00adsulfon\u00adyl)phen\u00adyl]-1,2,4-oxa\u00addiazol-5-yl}phen\u00adoxy)acetamide -N-(methyl\u00adsulfon\u00adyl)acetamide -N-(methyl\u00adsulfon\u00adyl)acetamide phen\u00adyl)acetamide -4-nitro\u00adphen\u00adyl]-N-methyl\u00adacetamide acetamide \u00adacetyl chloride were added. The mixture was stirred at room temperature for 5\u2005h. After this, 100\u2005ml of water were added and the mixture was extracted three times, each with 100\u2005ml of methyl tert-butyl ether (MTBE). The combined organic phases were dried over MgSO4 and the solvent was evaporated under reduced pressure. The crude product, N-(4-fluoro\u00adphen\u00adyl)-N-isopropyl-2-(methyl\u00adthio)\u00adacetamide, was used for the next stage with purification (7.5\u2005g).In a 250\u2005ml flask (with a nitro\u00adgen inlet and a septum) was placed 5\u2005g of 4-fluoro-N-(4-fluoro\u00adphen\u00adyl)-N-isopropyl-2-(methyl\u00adthio)\u00adacetamide dissolved in 150\u2005ml of di\u00adchloro\u00admethane. After cooling to 263\u2013273\u2005K, 13.37\u2005g of meta-chloro\u00adperbenzoic acid in 100\u2005ml di\u00adchloro\u00admethane was added slowly at the same temperature. The mixture was stirred at room temperature for 5\u2005h. After this, 200\u2005ml of water were added and the organic layer was separated, and washed with 100\u2005ml of 10% sodium bicarbonate twice. The organic phases were dried over MgSO4 and the solvent was evaporated under reduced pressure. The crude product was purified by chromatography over SiO2 (hexa\u00adne:ethyl acetate 9:1 v/v). The title compound was recrystallized from diethyl ether solution in the form of colorless plates. The overall reaction scheme is shown in Fig.\u00a04To a 250\u2005ml round-bottomed flask (with a nitro\u00adgen inlet and a septum) was added 7.5\u2005g of 1H NMR: CDCl3 : 1.097\u20131.08 ; 3.198 ; 3.664 ; 5.006\u20134.938 ; 7.273\u20137.132 . MS m/z: 273.45 (M)+.6.sp2H), 0.98\u2005\u00c5 (RCH3), 0.99\u2005\u00c5 (R2CH2) and 1.00\u2005\u00c5 (R3CH). Uiso(H) parameters were set to values of either 1.2Ueq or 1.5Ueq (RCH3 only) of the attached atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023003675/hb8062sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989023003675/hb8062Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023003675/hb8062Isup3.cmlSupporting information file. DOI: 2258160CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The capsid p24 (CA-p24) antigen is a component of the viral capsid of human immunodeficiency virus (HIV) that has been commonly used for clinical diagnosis and monitoring of HIV infections in Enzyme-linked Immunosorbent Assays (ELISAs). Commercial CA-p24 ELISAs are widely used in research settings, but these kits are costly and have limited breadth for detecting diverse HIV isolates.Commercial CA-p24 antibodies were used as capture and detection antibodies. Specific CA-p24 ELISAs were established with these antibodies and tested for the detection of HIV-1 isolates with the aim of developing in-house protocols to recognize HIV-1 infections in vitro for research purposes.Here we present four protocols for in-house ELISAs to detect HIV CA-p24 using commercial antibodies. The assays were able to detect the CA-p24 antigen of different HIV-1 isolates tested. Comparison between the protocols showed that these in-house ELISAs exhibit high specificity, sensitivity, and reproducibility for CA-p24 quantitation but their reactivity varied per HIV-1 isolate and subtype.These optimized ELISA protocols represent valuable tools to investigate HIV-1 infections in research facilities at a lower price than commercial CA-p24 kits.The online version contains supplementary material available at 10.1186/s12985-023-02242-5. HIV infections are prevalent worldwide. If left untreated, HIV can progress to AIDS (Acquired Immunodeficiency Syndrome) and cause more than 600,000 deaths every year . The curCurrent research approaches include viral genome editing through CRISPR-Cas systems, stem cell transplants, and mRNA vaccines \u20137. SucceIn this study, we aimed to optimize and compare four in-house CA-p24 ELISA systems to detect and quantitate distinct HIV-1 isolates. We determined the reproducibility and sensitivity of the ELISA systems and described their characteristics for testing CA-p24 antigen.q\u2009=\u20090.0065, Kruskal\u2013Wallis\u2019 test), and 45\u2009\u00d7\u2009lower than very high-cost commercial kits (q\u2009=\u20090.0048) , Anogen (ANG), Sino Biological (SB), and R&D Systems (RND). The costs per each in-house assay range between \u20ac25 and \u20ac30 per one 96-well plate, which is 15\u2009\u00d7\u2009lower than medium-cost commercial kits, 25\u2009\u00d7\u2009lower than high-cost commercial kits , possibly due to the use of the same detection antibody. Similarly, SB-CA-p24 ELISA showed a significantly high correlation with ABR-CA-p24 ELISA . RND-CA-p24 ELISA also exhibited a significant high correlation with ABR-CA-p24 ELISA , suggesting a similar sensitivity between both assays, possibly due to their similar reactivity for HIV-1 B subtypes. The lowest correlations were observed between RND- and SB-CA-p24 ELISAs and between ANG- and SB-CA-p24 ELISAs , suggesting a high between-assay reproducibility for the ELISA systems.We then assessed the reproducibility of the ELISA systems, examining both their consistency within individual assays and their reliability between different assays. In order to assess within-assay reproducibility, we employed the NL4-3 HIV-1 isolate as a test model. We quantified its concentration in each assay by utilizing technical replicates and applying different inactivation treatments, including heat and/or detergent. We observed that the concentration of the NL4-3 isolate remained consistent between the replicates within the tested conditions guidelines classify HIV-1 as a Risk Group 3 agent. Research involving HIV-1 was approved by the institutional biosafety office and carried out according to laboratory biosafety 3 guidelines.Twenty-two different HIV-1 isolates were produced according to standard protocols . The HIVVirus stocks were generated on CD8\u2009+\u2009-depleted human peripheral blood mononuclear cells (PBMCs) for each isolate, including the lab-adapted strains, and virus production was measured by CA-p24 ELISA (in-house assays). The HIV stocks were inactivated with 0.1% Empigen followed by incubation at 56\u00a0\u00b0C for 30\u00a0min.Aalto Bio Reagents (ABR) system2O. Adjust pH to 7.4 with HCl and add ddH2O up to a volume of 1,000\u00a0mL. Store at room temperature (20\u201325\u00a0\u00b0C). Dilute 1:10 with ddH2O for 1\u2009\u00d7\u2009TBS buffer.10\u2009\u00d7\u2009Tris buffered saline (TBS) buffer: 1.37\u00a0M NaCl (80\u00a0g)\u2009+\u200927\u00a0mM KCl (2\u00a0g)\u2009+\u20092.4\u00a0g of Tris in a volume of 950\u00a0ml of ddHFirst wash buffer solution and dilution buffer:1\u2009\u00d7\u2009TBS\u2009+\u20090.05% Empigen: Add 83\u00a0ml of 30% Empigen into 500\u00a0ml of 1\u2009\u00d7\u2009TBS buffer (Table 2O. Adjust pH to 7.4 with HCl up to 1000\u00a0mL and purify with a 0.2\u00a0\u00b5m filter and store at room temperature (20\u201325\u00a0\u00b0C). Dilute 1:10 with ddH2O for 1\u2009\u00d7\u2009PBS buffer.10\u2009\u00d7\u2009phosphate-buffered saline (PBS): 20 PBS tablets and up to 950\u00a0mL of ddHSecond wash buffer solution:2O for 1\u2009\u00d7\u2009PBS buffer\u2009+\u20090.1% Tween.10\u2009\u00d7\u2009PBS buffer\u2009+\u20091% Tween. Add 10\u00a0ml of Tween 20 into 990\u00a0ml of 10\u2009\u00d7\u2009PBS buffer. Store at room temperature (20\u201325\u00a0\u00b0C). Dilute 1:10 with ddH3: 4.2\u00a0g NaHCO3 and up to 450\u00a0mL of ddH2O. Adjust pH to 8.5 and add ddH2O up to a volume of 500\u00a0mL. Store at room temperature (20\u201325\u00a0\u00b0C).0.05\u00a0M NaHCOCoating buffer and antibody:3 (10\u00a0\u00b5g/mL) and dispense 25\u00a0\u00b5l of capture antibody solution into each well.Anti-HIV Gag CA-p24: reconstitute according to manufacturer\u2019s instructions. Make 100\u00a0\u00b5l aliquots and store at \u2212\u00a020\u00a0\u00b0C until use. To coat one 96-well plate, add 25\u00a0\u00b5l of aliquoted capture antibody into 2,475\u00a0\u00b5l of 0.05\u00a0M NaHCOCA-p24 protein: reconstitute with 1\u2009\u00d7\u2009TBS, 20% Sheep serum, and 1% Empigen to a concentration of 10\u00a0\u00b5g/mL. Dilute 1:10 with 1\u2009\u00d7\u2009TBS\u2009+\u20091% Empigen to a concentration of 1\u00a0\u00b5g/mL. Make 30\u00a0\u00b5l aliquots and store at -80\u00a0\u00b0C until use.CA-p24 standard curve: add 25\u00a0\u00b5l of aliquoted CA-p24 standard into 225\u00a0\u00b5l of dilution buffer (1\u2009\u00d7\u2009TBS\u2009+\u20090.05% Empigen) (working stock). Prepare standard curve dilutions as specified in Additional File CA-p24 standard protein and curve:ABR, ANG):HIV-CA-p24, Alkaline Phosphatase Conjugate monoclonal antibody (AP-mAb): reconstitute and dilute 1:5 by adding 20\u00a0\u00b5l of antibody into 80\u00a0\u00b5l triethanolamine. Store at 4\u00a0\u00b0C until use.Prepare the conjugate for one 96-well plate as followed: 2\u00a0mL of 1\u2009\u00d7\u2009TBS\u2009+\u20090.5\u00a0mL of sheep serum\u2009+\u20092.5\u00a0\u00b5l of Tween 20\u2009+\u20090.05\u00a0g of skim milk (2%)\u2009+\u20090.31\u00a0\u00b5l of the reconstituted conjugated . Dispense 25\u00a0\u00b5l of conjugate solution into each well.Detection antibody, conjugate, and substrate into each well Table .2.Sino Biological (SB) and Anogen (ANG) systemsABR protocol). Store at room temperature (20\u201325\u00a0\u00b0C). Dilute 1:10 with ddH2O for 1\u2009\u00d7\u2009PBS buffer\u2009+\u20090.05% Tween : to prepare the blocking buffer needed for one 96-well plate, add 0.2\u00a0g of BSA into 10\u00a0ml of 1\u2009\u00d7\u2009PBS buffer\u2009+\u20090.05% Tween.1\u2009\u00d7\u2009PBS\u2009+\u20090.05% Tween\u2009+\u20090.1% BSA: to prepare the dilution buffer needed for one 96-well plate, dilute 1:20 the blocking buffer into new 1\u2009\u00d7\u2009PBS buffer\u2009+\u20090.05% Tween as needed.SB): reconstitute according to manufacturer\u2019s instructions. To coat one 96-well plate, add 5.6\u00a0\u00b5l of coating antibody into 2.8\u00a0mL of 1\u2009\u00d7\u2009PBS (2\u00a0\u00b5g/mL) and dispense 25\u00a0\u00b5l of coating antibody solution into each well (Table Mouse Anti-HIV-CA-p24 (Coating antibodies:ANG): reconstitute according to manufacturer\u2019s instructions. To coat one 96-well plate, add 25\u00a0\u00b5l of aliquoted coating antibody into 2,475\u00a0\u00b5l of 1\u2009\u00d7\u2009PBS (10\u00a0\u00b5g/mL) and dispense 25\u00a0\u00b5l of coating antibody solution into each well into 225\u00a0\u00b5l of dilution buffer (1\u2009\u00d7\u2009PBS\u2009+\u20090.05% Tween\u2009+\u20090.1% BSA) (working stock). Prepare standard curve dilutions as specified in Additional File CA-p24 standard curve:SB): reconstitute according to manufacturer\u2019s instructions. Dilute 1:1,000 in HRP stabilizer (Table Mouse Anti-HIV-CA-p24 horseradish peroxidase (HRP) antibody (SB): to prepare the conjugate for one 96-well plate, dilute 1:1,000 aliquoted detection antibody by adding 3\u00a0\u00b5l of antibody into 3,000\u00a0\u00b5l of dilution buffer (100\u00a0ng/mL) and dispense 25\u00a0\u00b5l of conjugate solution into each well.Conjugate (SB): dilute 1:10 Lumiphos A and B solutions by adding 150\u00a0\u00b5l of solution A and 150\u00a0\u00b5l of solution B into 2.7\u00a0ml of ddH2O. Dispense 25\u00a0\u00b5l of LumiPhos A\u2009+\u2009B solution into each well SB/ANG protocols) with ddH2O.1\u2009\u00d7\u2009PBS buffer\u2009+\u20090.05% Tween: dilute 1:10 10\u2009\u00d7\u2009PBS buffer\u2009+\u20090.5% Tween into 45\u00a0\u00b5l of dilution buffer (1\u2009\u00d7\u2009PBS\u2009+\u20090.2% TritonX-100\u2009+\u20091% BSA) (working stock). Prepare standard curve dilutions as specified in Additional File CA-p24 standard curve: add 5\u00a0\u00b5l of aliquoted CA-p24 standard (CA-p24 standard curve:Biotinylated Mouse Anti-HIV-1 Gag CA-p24 Detection Antibody: reconstitute according to manufacturer\u2019s instructions with 1.0\u00a0ml of dilution buffer (1\u2009\u00d7\u2009PBS\u2009+\u20090.2% TritonX-100\u2009+\u20091% BSA). Make 45\u00a0\u00b5l aliquots and store at \u2212\u00a020\u00a0\u00b0C until use. To prepare the antibody solution for one 96-well plate, add 41.8\u00a0\u00b5l of aliquoted detection antibody into 5\u00a0mL of 1\u2009\u00d7\u2009PBS\u2009+\u20090.2% TritonX-100\u2009+\u20091% BSA buffer (125\u00a0ng/mL) and dispense 50\u00a0\u00b5l of detection antibody solution into each well.Conjugate: to conjugate the detection antibody in one 96-well plate, dilute 1:80 Streptavidin-HRP solution by adding 62.5\u00a0\u00b5l of antibody into 5\u00a0ml of 1\u2009\u00d7\u2009PBS\u2009+\u20090.2% TritonX-100\u2009+\u20091% BSA buffer and dispense 50\u00a0\u00b5l of solution into each well.2O was used to analyze datasets and determine the sensitivity of the assays and Pearson\u2019s Additional file 1: Supplementary Figs.\u00a01\u20134. (.pdf). Fig. S1. Comparison of costs between in-house and commercial ELISA kits. Fig. S2. Standard curves generated by ABR-, ANG-, SB-, and RND-CA-p24 ELISAs. Fig. S3. Effect of viral inactivation on assay performance and within-assay reproducibility of ELISA systems. Fig. S4. Between-assay reproducibility of the ELISA systems.Additional file 2: Supplementary Tables 1\u20134 (.pdf). Table S1. List of commercial ELISA kits and their costs. Tables S2\u2013S4. Four tables with the volumes and details needed to prepare the standard curves for the ABR-, ANG-, SB-, and RND-CA-p24 ELISAs."} +{"text": "A short and directional tetrel bond between tin and oxygen is identified in the title compound. 6H5)3]\u00b7C18H21OP, is reported. The 1:1 cocrystal features a short and directional tetrel bond between tin and oxygen. The tin\u2013oxygen distance is 2.346\u2005(4)\u2005\u00c5, representing 62% of the sum of the van der Waals radii of Sn and O. The Cl\u2014Sn\u22efO angle is 174.0\u2005(1)\u00b0 and this nearly linear arrangement is consistent with a tetrel bond formed via a \u03c3-hole opposite the tin\u2013chlorine covalent bond. Some weak C\u2014H\u22efCl inter\u00adactions are noted between adjacent mol\u00adecules.The single-crystal X-ray diffraction structure of the title compound, [SnCl(C The asymmetric unit consists of one complete mol\u00adecule of each type. The tin\u2013oxygen distance is 2.346\u2005(4)\u2005\u00c5 and the Cl\u2014Sn\u22efO TB angle is 174.0\u2005(1)\u00b0 \u2005\u00c5 and the Cl\u2014Sn\u22efO TB angle is 177.57\u2005(7)\u00b0 (p-tol\u00adyl)tin]-\u03bc-1,2-bis\u00ad(di\u00adphenyl\u00adphosphor\u00adyl)ethane-\u03ba2O:O\u2032-[bromido\u00adbis\u00ad(p-chloro\u00adphen\u00adyl)(p-tol\u00adyl)tin] 2\u00b7(Me2N)2PO tin\u2013hexa\u00admethyl\u00adphospho\u00adr\u00adamide , a moderately strong and directional noncovalent inter\u00adaction, has received renewed inter\u00adest in recent years as a useful structure-directing element and crystal engineering tool \u00b0 Fig.\u00a01. This divia slow evaporation of the solvent in a fume hood over a period of 5\u2005d. Evidently, during the synthesis, the phosphane was oxidized to give the phosphane oxide, as the process was not carried out under an inert atmosphere.In a typical procedure, triphenyltin(IV) chloride (0.0614\u2005g) and cyclo\u00adhexyl\u00addiphenyl\u00adphosphane (0.0894\u2005g) were added to hexane (60\u2005ml) in a beaker. The mixture was heated and stirred until the solids were completely dissolved. Cocrystals grew The crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314623006375/bv4048sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314623006375/bv4048Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623006375/bv4048Isup3.cmlSupporting information file. DOI: 2267964CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Hirshfeld surface analysis indicates that H\u22efH and H\u22efC/C\u22efH contacts are the most important inter\u00adactions.A new copper(II) complex with 3-methyl\u00adbenzoate and 2,2\u2032-bi\u00adpyridine synthesized displays chains of hydrogen-bonded complex units along the 2N,N\u2032)bis\u00ad(3-methyl\u00adbenzoato)-\u03ba2O,O\u2032;\u03baO-copper(II) 0.68-hydrate, [Cu(C8H7O2)2(C10H8N2)(H2O)]\u00b70.68H2O or [Cu(3-mb)2(bipy)(H2O)]\u00b70.68H2O. The coord\u00adination environment of CuII is a distorted octa\u00adhedron. The metal atom is attached to two 3-mb moieties, which bind in monodentate and bidentate fashions. One of the 3-mb units is disordered. The coordination environment is completed by one bipy ligand and a water mol\u00adecule. A second water mol\u00adecule is outside the coordination sphere of the CuII atom and its occupancy refined to 0.68. The structure consists of chains along the b-axis direction formed by complex units joined via hydrogen bonds between the coordinated water mol\u00adecule and an O atom of a coordinated 3-mb unit. Hirshfeld surface analysis indicates that the most abundant contacts are H\u22efH (56.8%), H\u22efC/C\u22efH (21.7%) and H\u22efO/O\u22efH (13.7%).3-Methyl\u00adbenzoic acid (3-mbH) and 2,2\u2032-bi\u00adpyridine (bipy) reacted with a cop\u00adper(II) salt forming a new mixed ligand complex, aqua\u00ad(2,2\u2032-bi\u00adpyridine-\u03ba The complex was octa\u00adhedral with 3-mb acting as monodentate complexes continues to be of inter\u00adest. Copper is an important part of various metalloenzymes. It takes part in many metabolic processes such as iron metabolism, mitochondrial oxidative phospho\u00adrylation and catecholamine production to 2.0131\u2005(18)\u2005\u00c5 and the N1\u2014Cu1\u2014N2 angle is 80.58\u2005(7)\u00b0 (Table\u00a01carboxyl\u00adate distances are 1.842\u2005(17)\u20132.2988\u2005(18)\u2005\u00c5. The Cu\u2014O and Cu\u2014N values are very close to those reported for copper(II) complexes involving benzoate (BZA) as a ligand, for example [Cu(BZA)2(bipy)(H2O)] [Cu\u2014O = 1.9951\u2005(12)\u20131.9633\u2005(12)\u2005\u00c5 and Cu\u2014N = 2.0064\u2005(14)\u20132.0111\u2005(13)\u2005\u00c5; Devereux et al., 2007Complex 1 Fig.\u00a01 crystall\u00b0 Table\u00a01. The Cu13.B\u22efO4) leads to the formation of a linear chain in the b-axis direction refined to 0.68, which seems to be due to water escaping the crystal through the channels that run along the b-axis direction.In the crystal, hydrogen bonding between H atoms of the coord\u00adin\u00adated water mol\u00adecule and the O atoms of the coordinated 3-mb , H\u22efC/C\u22efH and H\u22efO/O\u22efH contacts are 56.8, 21.7 and 13.7%, respectively.\u22efH Fig.\u00a04c and H\u22ef\u22efH Fig.\u00a04d contac6.3)2\u00b73H2O in water (20\u2005ml) under stirring. A solution of 2,2\u2032-bi\u00adpyridine in EtOH (25\u2005ml) was added and the color changed from greenish blue to blue. The precipitate was filtered off, washed with water and dried. Blue single crystals of the title complex suitable for X-ray diffraction studies were obtained after evaporation of an ethanol solution after several days.3-Methyl\u00adbenzoic acid and sodium hydroxide in water (20\u2005ml) were added to a solution of Cu(NO7.A\u2013O6\u2013H6B refined to 0.680\u2005(10). The coordinates of the ordered water atom were refined with Uiso(H) = 1.5Ueq(O). All other H atoms were positioned geometrically and refined as riding with Uiso(H) = 1.2\u20131.5Ueq(parent atom).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023006904/dj2063sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023006904/dj2063Isup2.hklStructure factors: contains datablock(s) I. DOI: 2117143CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The formation of a co-crystal sustained by \u03c0-type halogen bonds involving 1,4-di\u00adiodo\u00adperchloro\u00adbenzene and naphthalene is reported. 6I2Cl4) as the halogen-bond donor along with naphthalene (nap) as the acceptor is reported. The co-crystal generates a chevron-like structure that is held together primarily by \u03c0-type halogen bonds (i.e. C\u2014I\u22ef\u03c0 contacts) between the components. In addition, C6I2Cl4 also inter\u00adacts with the acceptor via C\u2014Cl\u22ef\u03c0 contacts that help stabilize the co-crystal. Within the solid, both aromatic components are found to engage in offset and homogeneous face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions. Lastly, the halogen-bond donor C6I2Cl4 is found to engage with neighboring donors by both Type I chlorine\u2013chlorine and Type II iodine\u2013chlorine contacts, which generates an extended structure.The formation and crystal structure of a co-crystal based upon 1,4-di\u00adiodo\u00adperchloro\u00adbenzene (C Recently, we reported the formation of photoreactive co-crystals based upon C6I2Cl4 along with trans-1,2-bis\u00ad(pyridine-4-yl)ethyl\u00adene resulting in a chevron-like structure. In addition to the \u03c0-type halogen bond, the co-crystal (C6I2Cl4)\u00b7(nap) is also held together by the combination of C\u2014Cl\u22ef\u03c0 contacts, homogeneous face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions, Type I chlorine\u2013chlorine contacts, and Type II iodine\u2013chlorine contacts.A continued goal within our research groups has been in the design and formation of halogen-bonded co-crystals based upon 1,4-di\u00adiodo\u00adperchloro\u00adbenzene (C2.6I2Cl4)\u00b7(nap) crystallizes in the centrosymmetric triclinic space group P\u012b. The asymmetric unit contains half a mol\u00adecule of both C6I2Cl4 and nap where inversion symmetry generates the remainder of each mol\u00adecule \u00b7(nap), the donor C6I2Cl4 is found to engage in Type I chlorine\u2013chlorine contacts 1\u00a0\u2212\u00a0x, -y, 1 \u2212 z], with a distance of 3.499\u2005(1)\u2005\u00c5 and a C\u2014Cl\u22efCl bond angle of \u03b81 = \u03b82 = 132.16\u2005(6)\u00b0 1\u00a0\u2212\u00a0x, -y, 1 \u2212 z], with a distance of 3.808\u2005(1)\u2005\u00c5 and a C\u2014I\u22efCl bond angle of 111.83\u2005(4)\u00b0. Both the aromatic halogen-bond donor and acceptor are found to engage in an offset and homogeneous face-to-face \u03c0\u2013\u03c0 stacking arrangement that stabilizes the co-crystal \u2005\u00c5 measured for Cl1\u22efC5.In addition to \u03c0-type halogen bond within \u00b7(nap). This Hirshfeld surface analysis along with the observed bond lengths confirms the ability of C6I2Cl4 to engage in \u03c0-type halogen bonds to a polycyclic aromatic hydro\u00adcarbon, namely nap.These various non-covalent inter\u00adactions were also investigated and visualized by utilizing a Hirshfeld surface analysis in which the I atom is within the van der Waals radius of an aromatic surface revealed only one structure, refcode HONBIY \u00b7(nap).A search of the Cambridge Crystallographic Database was synthesized utilizing a previously published method were both purchased from Sigma-Aldrich Chemical and used without any additional purification. The halogen-bond donor 1,4-di\u00adiodo\u00adperchloro\u00adbenzene \u00b7(nap) was achieved by dissolving 50.0\u2005mg of C6I2Cl4 in 2.0\u2005mL of toluene and then combined with a 2.0\u2005mL toluene solution containing 13.7\u2005mg of nap . Within two days, single crystals suitable for X-ray diffraction were formed upon loss of some of the solvent by slow evaporation.The formation of I. DOI: 10.1107/S2056989023008356/jy2037Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023008356/jy2037Isup3.cmlSupporting information file. DOI: 2291675CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title com\u00adplex, bis\u00ad{\u03bc-1,1\u2032-(o-xylylene)bis\u00ad[4-(pyridin-2-yl)triazole]-\u03baN3:N3\u2032}dipalladi\u00adum(II) tetra\u00adkis\u00ad(tetrafluoridoborate)\u2013di\u00admethyl\u00adformamide\u2013diethyl ether (1/2/1), as the BF4 salt, and including di\u00admethyl\u00adformamide and diethyl ether solvent mol\u00adecules, with stoichiometry [Pd2(C22H18N8)2](BF4)4\u00b72C3H7NO\u00b7C4H10O, the Pd complex and the disordered diethyl ether mol\u00adecule lie on inversion centers. The ligand coordinates to the PdII centers with square-planar geometry, forming a dimeric macrocycle. The Pd\u22efPd separation in the complex [Pd2(o-xpt)2]4+ cation is 3.6184\u2005(4)\u2005\u00c5. In the crystal, the complex mol\u00adecules are stacked along the b axis, with \u03c0\u2013\u03c0 inter\u00adactions between the pyridyl\u00adtriazole ligands of two mol\u00adecules.A tetra\u00addentate ligand, namely, Herein, we report the synthesis and crystal structure of the title compound.The self-assembly of polydentate ligands with transition-metal ions to create functional metal\u2013organic supra\u00admolecules has been of great inter\u00adest in recent years. In particular, the complexation of 2-pyridyl-1,2,3-triazole chelating pockets with transition-metal ions has intensified due to the ease of synthesis of the ligands through the copper-catalyzed azide\u2013alkyne cyclo\u00adaddition (CuAAC) reaction 2]4+ lying on an inversion center, two BF4\u2212 anions in general positions, a di\u00admethyl\u00adformamide solvent mol\u00adecule in a general position and a diethyl ether solvent mol\u00adecule disordered about an inversion center atoms of two pyridyl\u00adtriazole moieties. The N\u2014Pd\u2014N chelating angles are N1\u2014Pd1\u2014N2 = 79.46\u2005(8)\u00b0 and N7\u2014Pd1\u2014N8 = 79.75\u2005(8)\u00b0. The N(py)\u2014Pd bonds are slightly longer than N(trz)\u2014Pd bonds , suggesting the triazolium N atom coordinates more strongly to the Pd center than the pyridyl N atom of the ligand. These values are typical for the reported mononuclear PdII complex of pyridyl\u00adtriazole ligands with an average interplanar distance of 3.364\u2005\u00c5, indicating \u03c0\u2013\u03c0 interaction between the mol\u00adecules; this is shown along the 3CN)4]BF4 in aceto\u00adnitrile (5\u2005ml), o-xpt in aceto\u00adnitrile (5\u2005ml) was added dropwise. The solution was stirred for 1\u2005h at room temperature. The volatiles were removed in vacuo. The residue was washed with di\u00adchloro\u00admethane (2\u2005ml), followed by methanol (2\u2005ml), and dried under vacuum to give [Pd2(o-xpt)2](BF4)4 as a pale-yellow solid. Crystals suitable for X-ray analysis were obtained by slow vapor diffusion of diethyl ether into a di\u00admethyl\u00adformamide (DMF) solution of the complex at room tem\u00adper\u00adature. Our attempts to obtain a clean 1H NMR spectrum in DMSO-d6 were not successful, possibly due to the labile nature of the complex in solution. High resolution ESI\u2013MS analysis showed a monocationic signal at m/z 1255.1478 {calculated 1255.1540 for [Pd2(o-xpt)2(BF4)4]+}.To a stirred solution of [Pd values were assigned as 1.2Ueq for the attached atom (1.5 for meth\u00adyl). A torsional parameter was refined for each methyl group, except for those of the diethyl ether mol\u00adecule, which were staggered with respect to CH2. The diethyl ether solvent mol\u00adecule is disordered about an inversion center with two half-populated sites. A number of distance and displacement parameter restraints were necessary to model the disorder.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2414314623003620/xu4050sup1.cifCrystal structure: contains datablock(s) I, glonal. DOI: 10.1107/S2414314623003620/xu4050Isup2.hklStructure factors: contains datablock(s) I. DOI: 2257865CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dihedral angles between the A and B rings and their attached phenyl rings are 49.15\u2005(8) and 80.78\u2005(8)\u00b0, respectively. In the extended structure, the mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions, which variously generate C(11) chains and Edis at \u2212428.6\u2005kJ\u2005mol\u22121 being the major contributor.In the title compound, C 38H28O6, the dihedral angles between the naphthalene ring system and its pendant benz\u00adyloxy rings A and B are 88.05\u2005(7) and 80.84\u2005(7)\u00b0, respectively. The dihedral angles between the A and B rings and their attached phenyl rings are 49.15\u2005(8) and 80.78\u2005(8)\u00b0, respectively. In the extended structure, the mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 hydrogen bonds, and \u03c0\u2013\u03c0 stacking inter\u00adactions, which variously generate C(11) chains and R22(12) loops as part of a three-dimensional network. The Hirshfeld surface and inter\u00admolecular inter\u00adaction energies are reported, with dispersion (Edis = \u2212428.6\u2005kJ\u2005mol\u22121) being the major contributor.In the title compound, C A variety of banana-shaped, bow-shaped or bent-core ferroelectric liquid crystals were developed by incorporating a benzene ring as a rigid core and its pendant C26\u2013C31 (A) and C12\u2013C17 (B) benz\u00adyloxy rings are 88.05\u2005(7) and 80.84\u2005(7)\u00b0, respectively. The dihedral angles between the A and B rings and their attached C33\u2013C38 and C19\u2013C24 phenyl rings are 49.15\u2005(8) and 80.78\u2005(8)\u00b0, respectively. Key torsion angles include C1\u2014O4\u2014C25\u2014C26 [\u2212160.98\u2005(13)\u00b0], C28\u2014O2\u2014C32\u2014C33 [\u2212172.04\u2005(14)\u00b0], C10\u2014O1\u2014C11\u2014C12 [\u2212168.94\u2005(14)\u00b0] and C14\u2014O3\u2014C18\u2014C19 [172.84\u2005(14)\u00b0]. Otherwise, the geometrical data for the title compound may be regarded as normal.The title compound crystallizes with one mol\u00adecule in the asym\u00admetric unit Fig.\u00a01 in the s3.C(11) chain (arising from the C21\u2014H21\u22efO2ii hydrogen bond), which runs along [010], and centrosymmetric i hydrogen bond) between the mol\u00adecules as shown in Fig.\u00a02In the crystal, the mol\u00adecules are linked by numerous C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions Table\u00a01. Promine4.et al., 2007CrystalExplorer17 are electrostatic (Eele), polarization (Epol), dispersion (Edisp) and exchange\u2013repulsion (Erep), and the cylinder-shaped energy frameworks represent the relative strengths of the inter\u00adaction energies in individual directions, as well as the topologies of pairwise inter\u00admolecular inter\u00adaction energies within the crystal in DCM (20\u2005ml). Filtration was used to remove the precipitated N,N-di\u00adcyclo\u00adhexyl\u00adurea and the solvent was evaporated. To obtain the pure product, the solid residue was purified using column chromatography on silica gel with DCM as an eluent, followed by recrystallization from ethyl alcohol solution.Under an inert atmosphere, 1,2-di\u00adhydroxy\u00adnaphthalene (1.00\u2005mmol), a catalytic amount of 4-di\u00admethyl\u00adamino\u00adpyridine and 3-benzyl\u00adoxybenzoic acid (2.00\u2005mmol) were dissolved in 50\u2005ml of dry di\u00adchloro\u00admethane (DCM). The above mixture was stirred for 2\u2005h at room temperature with a solution of 7.Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023005571/hb8069sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989023005571/hb8069Isup4.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023005571/hb8069sup3.docxSupplementary Figures showing C--H...pi interactions. DOI: 2271880CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Compounds (I)\u2013(III) are constructed from vertex-sharing ZnO3N tetra\u00adhedra (the organic mol\u00adecule acting as a ligand) and HPO3 pseudo pyramids in a 1:1 ratio to generate the same motif of infinite 4-ring \u2018ladder\u2019 chains propagating in the [010], [101] and [100] directions, respectively, whereas (IV) consists of (010) layers of vertex-sharing ZnO4 and HPO3 units in a 3:4 ratio with the protonated organic mol\u00adecule acting as a template. When an excess of HCl is used in the synthesis, the simple hydrated mol\u00adecular salt, bis\u00ad(2-amino-3-methyl\u00adpyridinium) tetra\u00adchloro\u00adzincate monohydrate, (C6H8N2)2[ZnCl4]\u00b7H2O, (V), arises. Com\u00adpounds (I)\u2013(V) feature extensive networks of hydrogen bonds, both classical and non-classical in nature, which help to consolidate the extended structures.The syntheses and crystal structures of four hydro\u00adthermally prepared organo\u2013zinc phosphites, Each O atom is bonded to one Zn and one P atom [mean Zn\u2014O\u2014P = 130.2\u00b0], thus there are no \u2018dangling\u2019 The asymmetric unit of (I)C2/c, consists of two Zn2+ ions, two [HPO3]2\u2013 hydrogen phosphite anions, and two C6H7N2 2-amino-4-methyl\u00adpyridine mol\u00adecules acting as ligands, i.e., Z\u2032 = 2. Unlike (I)3N tetra\u00adhedra and [HPO3]2\u2013 dianions \u2013 and the major structural features of (II)4\u2032 = 0.97; comparable data for Zn2 = 1.940\u2005\u00c5, 100.10\u2005(5)\u2013121.28\u2005(5)\u00b0 and 0.91, respectively; mean P1\u2014O = 1.525\u2005\u00c5; spread of O\u2014P1\u2014O bond angles = 110.64\u2005(6)\u2013113.78\u2005(6)\u00b0; P1 displacement from its attached O atoms = 0.4278\u2005(7)\u2005\u00c5; comparable data for P2 = 1.520\u2005\u00c5, 111.44\u2005(6)\u2013113.14\u2005(7)\u00b0 and \u22120.4269\u2005(8)\u2005\u00c5, respectively. All six O atoms are bridging between Zn and P atoms with a mean bond angle of 131.3\u00b0 [range = 123.26\u2005(6)\u2013144.09\u2005(8)\u00b0]. The extended structure of (II)The asymmetric unit of (II)P212121 with a well-defined absolute structure and its asymmetric unit and 115.09\u2005(8)\u00b0, respectively, \u03c44\u2032 = 0.96] and [HPO3]2\u2013 units . The three O atoms bridge adjacent zinc and phospho\u00adrus atoms with a mean Zn\u2014O\u2014P bond angle of 126.7\u00b0. For the extended structure of (III)Compound (III)it Fig.\u00a03 consistsP4 tetra\u00adhedra and four [HPO32\u2013] pseudo pyramids as well as two protonated 2-amino-4-methyl\u00adpyridinium cations, which therefore act as templates rather than ligands; a water mol\u00adecule of crystallization (O13) completes the structure. Geometrical data for the zinc polyhedra are as follows: mean Zn1\u2014O = 1.941\u2005\u00c5, spread of bond angles = 100.42\u2005(8)\u2013122.18\u2005(9)\u00b0, \u03c44\u2019 = 0.90; equivalent data for Zn2: 1.936\u2005\u00c5, 98.33\u2005(8)\u2013115.30\u2005(9)\u00b0 and 0.98, respectively; equivalent data for Zn3: 1.945\u2005\u00c5, 99.70\u2005(8)\u2013117.10\u2005(8)\u00b0 and 0.96, respectively. The four [HPO3]2\u2013 anions adopt their normal geometries: mean P1\u2014O = 1.519\u2005\u00c5, minimum and maximum O\u2014P1\u2014O = 111.00\u2005(11) and 112.70\u2005(11)\u00b0, respectively, deviation of P1 from its attached O atoms = 0.4498\u2005(13)\u2005\u00c5; equivalent data for P2: 1.522\u2005\u00c5, 110.08\u2005(10)\u00b0, 115.33\u2005(11)\u00b0 and \u22120.4122\u2005(12)\u2005\u00c5, respectively; equivalent data for P3: 1.516\u2005\u00c5, 110.19\u2005(11)\u00b0, 114.49\u2005(12)\u00b0 and \u22120.4123\u2005(13)\u2005\u00c5, respectively; equivalent data for P4: 1.516\u2005\u00c5, 112.68\u2005(12)\u00b0, 114.13\u2005(12)\u00b0 and 0.3903\u2005(13)\u2005\u00c5, respectively. The twelve unique O atoms all bridge Zn and P atoms , maximum = 146.90\u2005(13), spread = 21.5\u00b0). For the extended structure of (IV)In (IV)it Fig.\u00a04 reveals P6H8N2+ cations protonated at their pyridine N atoms, a [ZnCl4]2\u2013 anion and a water mol\u00adecule of crystallisation. The tetra\u00adchloro\u00adzincate ion has a mean Zn\u2014Cl separation of 2.2704\u2005\u00c5 [range = 2.2536\u2005(13)\u20132.2867\u2005(13)\u2005\u00c5] and smallest and largest Cl\u2014Zn\u2014Cl bond angles of 104.48\u2005(5) and 113.75\u2005(5)\u00b0, respectively. The synthetic intent here was to lower the pH with HCl and establish if a di\u00adhydrogen phosphite (H2PO3\u2212) anion containing a terminal P\u2014OH moiety could be incorporated into the structure 3.3N and HPO3 polyhedra are linked by Zn\u2014O\u2014P bonds into [010] polyhedral 4-ring (two Zn and two P nodes) \u2018ladder\u2019 chains in which the zinc and phospho\u00adrus nodes strictly alternate to adjacent P atoms (and a fourth bond to the organic species) and that the P atom forms three links to zinc atoms (and a fourth P\u2014H vertex), the 1:1 Zn:P stoichiometry is to be expected and hence no charge compensating, protonated template is needed. In (II)Z\u2032 = 2, every other 4-ring is generated by inversion symmetry and translation in the [101] direction leads to the extended array. In (III)motif is again apparent te Fig.\u00a06: the chant Fig.\u00a06, althougns Fig.\u00a06.4 and HPO3 polyhedra sharing corners. One way to visualize this rather complex arrangement is in terms of contorted chains of 4-rings featuring atoms Zn1, Zn2, Zn3, P2, P3 and P4 as the nodes propagating in the [001] direction. One out of every three 4-rings in a chain is generated by inversion symmetry. These chains are cross-linked in the a-axis direction by the P1-centred hydrogen phosphite groups to form the (010) layers, which encapsulate 8-ring voids built up from four Zn and four P nodes although there is no suggestion of \u2018zeolitic\u2019 porosity. So far as stoichiometry is concerned, in this case the zinc nodes forming four bonds to nearby phospho\u00adrus atoms and the P nodes forming three bonds to Zn atoms leads to the 3:4 ratio of zinc and phospho\u00adrus, which is the proportion most commonly seen in this family of phases 4] unit, hence the two protonated template mol\u00adecules. The template cations and water mol\u00adecules of crystallisation occupy the inter-layer regions.The extended structure of (IV)Various classical and non-classical (C\u2014H\u22efO and C\u2014H\u22efCl) hydrogen bonds occur in these structures. As is normal, the hydrogen phosphite P\u2014H unit does not participate in hydrogen bonding and inter-chain (via H1N and H3N) links, with the latter serving to cross-link the [101] chains into a three-dimensional network (Table\u00a02B\u22efO2 bond (Table\u00a03A) of the amine grouping does not participate in a hydrogen bond, the closest acceptor O atom being some 2.77\u2005\u00c5 distant. There are no significant \u03c0\u2013\u03c0 stacking inter\u00adactions in (III)In (II)k Table\u00a02. The arok Table\u00a02, the sind Table\u00a03 cross-livia two O\u2014H\u22efO hydrogen bonds. The N\u2014H\u22efO hydrogen bonds originating from the protonated pyridine N atoms and the \u2013NH2 groups of the organic species all link to the same sheet for each template cation, i.e., there are no inter-sheet hydrogen bonds associated with the templates. Significant aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions occur between centrosymmetric pairs of each template cation, as indicated by the centroid\u2013centroid separation of 3.6167\u2005(15)\u2005\u00c5 (slippage = 1.196\u2005\u00c5) for the C1 species and 3.4695\u2005(17)\u2005\u00c5 (0.146\u2005\u00c5) for the C7 cation.In (IV)w (w = water) and O\u2014H\u22efCl inter\u00adactions to generate [001] chains; within these chains, centrosymmetric assemblages of two C6H8N2+ cations, two [ZnCl4]2\u2013 anions and two water mol\u00adecules are apparent t Fig.\u00a012.4.et al., 20165H6N2\u00b7Zn(HPO3)]n, catena-[(\u03bc3-hydrogenphosphito)(2-amino\u00adpyridine)\u00adzinc] (CSD refcode LUZYOU) layers of 4- and 8-rings.A survey of the Cambridge Structural Database (Groom 3 units in (I)motif in the wider ZnPO phase space: two other examples with very different ligating mol\u00adecules to those in (I)\u2013(III) are [C4H8N2O3\u00b7Zn(HPO3)]n (C4H8N2O3 = l-asparagine) ]n \u2013(III).The fact that the N-bonded zinc ions and HPO6H8N2 organic mol\u00adecule acts as a ligand (a Zn\u2014N bond and a 1:1 Zn:P ratio) and (IV)6H9N2+ template (N\u2014H\u22efO hydrogen bonds and a 3:4 Zn:P ratio) arose from similar syntheses, with the only difference being the source of zinc ions . Assuming that hydro\u00adthermal synthesis is not just an impenetrable \u2018black box\u2019 5.3PO3 and 1.14\u2005g of 2-amino-3-methyl\u00adpyridine (Zn:P:template ratio \u2243 1:1:1), which were placed in a 50\u2005ml polypropyl\u00adene bottle with 20\u2005ml of water and shaken well to result in a white slurry. The bottle was placed in an 353\u2005K oven for 24\u2005h and then removed and allowed to cool to room temperature. The solids were recovered by vacuum filtration to result in a mass of needle-like transparent crystals. IR: 2383\u2005cm\u22121 (P\u2014H stretch). Increasing the heating time to one week led to the same product, with a slight improvement in crystallinity, as indicated by sharper peaks in its IR spectrum and X-ray powder diffraction pattern.Compound (I)3PO3 and 1.10\u2005g of 2-amino-4-methyl\u00adpyridine (Zn:P:template ratio \u2243 1:1:1); otherwise following the same procedure as for (I)\u22121 (P\u2014H stretch). Two peaks may arise because of the two different P\u2014H groups in the asymmetric unit 2, 0.86\u2005g of H3PO3 and 1.09\u2005g of 2-amino-5-methyl\u00adpyridine (Zn:P:template ratio \u2243 1:1:1) and 20\u2005ml of water were placed in a 50 ml polypropyl\u00adene bottle and heated to 353\u2005K for three days. Upon cooling, the product consisted of a mass of colourless blocks. IR: 2406\u2005cm\u22121 (P\u2014H stretch).To prepare compound (III)2, 0.77\u2005g of H3PO3 and 1.03\u2005g of 2-amino-4-methyl\u00adpyridine (Zn:P:template ratio \u2243 1:1:1) and 20\u2005ml of water. These components were placed in a 50-ml polypropyl\u00adene bottle and heated to 353\u2005K for 24\u2005h. Upon cooling, the product consisted of a mass of colourless blocks. IR: 3000\u20133600 (broad) (O\u2014H stretch), 2391, 2381\u2005cm\u22121 (P\u2014H stretch). The same product arises if the mixture is heated for one week.Compound (IV)M HCl.Compound (V)6.Uiso(H) = 1.2Ueq(N or O). The phosphite H atoms were geometrically placed (P\u2014H = 1.32\u2005\u00c5) and refined as riding atoms with Uiso(H) = 1.2Ueq(P). All the C-bound H atoms were located geometrically (C\u2014H = 0.95\u20130.98\u2005\u00c5) and refined as riding atoms with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C). The methyl groups were allowed to rotate, but not to tip, to best fit the electron density. Two peaks greater than 1\u2005e\u2005\u00c5\u22123 were found in the final difference map for (IV)Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989023002062/pk2681sup1.cifCrystal structure: contains datablock(s) I, II, III, IV, V, global. DOI: 10.1107/S2056989023002062/pk2681Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989023002062/pk2681IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989023002062/pk2681IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 10.1107/S2056989023002062/pk2681IVsup5.hklStructure factors: contains datablock(s) IV. DOI: 10.1107/S2056989023002062/pk2681Vsup6.hklStructure factors: contains datablock(s) V. DOI: 2246111, 2246110, 2246109, 2246108, 2246107CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Speciation, mechanism and polymer chain growth studies support a step\u2010growth process where reversible chain transfer occurs, i.e. H3B\u2009\u22c5\u2009PRH2/oligomer/polymer can all coordinate with, and be activated by, the catalyst. Block copolymer [H2BPPhH]110\u2010b\u2010[H2BP(n\u2010hexyl)H]11 can be synthesized and self\u2010assembles in solution to form either rod\u2010like micelles or vesicles depending on solvent polarity.An amphiphilic block copolymer of polyphosphinoborane has been prepared by a mechanism\u2010led strategy of the sequential catalytic dehydropolymerization of precursor monomers, H 2BPPhH]110\u2010b\u2010[H2BP(n\u2010hexyl)H]11, have been prepared by a mechanism\u2010led strategy of the catalytic reversible chain transfer dehydropolymerization of precursor monomers, H3B\u2009\u22c5\u2009PRH2 . These self\u2010assemble in THF:hexane solutions to form nanostructured rod\u2010like micelles, the morphology of which can be controlled by solvent polarity.Amphiphilic block copolymers of polyphosphinoboranes, [H Organic block copolymers (BCPs), in which chemically\u2010distinct segments of monomer units are linked together in a polymer chain, have played an important role in the development of macromolecular science.3, 4 T2BPRH]n (ClCH2Cl)][BArF4], I,2OTf, II.[I.II a non\u2010living chain\u2010growth mechanism is proposed.2PCH2CH2PPh2)2]Cl, 1, and harnessing a step\u2010growth like mechanism in which reversible chain transfer also occurs,Polyphosphinoborane homopolymers are generally (\u03b76\u2010FC6H5)[BArF4] (L1=Ph2PCH2CH2CH2PPh2) will dehydropolymerize H3B\u2009\u22c5\u2009PPhH2 under melt conditions to form [H2BPPhH]n, while solution conditions produced shorter\u2010chain oligomers.III complex with a chelating P\u2212H activated diboraphosphine is a plausible intermediate, e.g. [Rh(L1)H][BArF4], III 2C6H3);[F4]\u2212 anion we explored the use of air\u2010stable [Rh(Ph2PCH2CH2PPh2)2]Cl, 1, in the anticipation that the chelating phosphine may become labile under the conditions of catalysis (\u2248100\u2009\u00b0C). Optimization of conditions showed that 1\u2005mol% 1, 1.25\u2005M H3B\u2009\u22c5\u2009PPhH2, heated at 100\u2009\u00b0C in toluene for 19\u2005hours in a sealed thick\u2010walled NMR tube, followed by precipitation into hexanes, afforded [H2BPPhH]n as a white solid in 86\u2009% isolated yield.Mn=26\u2009500\u2005g\u2009mol\u22121, \u00d0=1.6 =355\u2005Hz. Under these conditions no significant amounts of shorter chain, or cyclic, oligomers are observed.[1 will also dehydropolymerize relatively electron\u2010poor H3B\u2009\u22c5\u2009P2C6H3)H2 to form higher molecular weight polyphosphinoborane [H2BP2C6H3)H]n, Mn=127\u2009500\u2005g\u2009mol\u22121, \u00d0=1.2 in the same reaction time .31P NMR spectrum by a statistical distribution of proposed rr, or mm triads, \u03b4(31P{1H}) \u221245.6, \u221246.9, \u221248.3 (1\u2009:\u20092\u2009:\u20091 respectively), as noted previously.[1 to form [H2BP(n\u2010hexyl)H]n as a colorless, hexane\u2010soluble, oil, Mn=33\u2009000\u2005g\u2009mol\u22121, \u00d0=1.3;,[2PCH2CH2PPh2)H]Cl, 2, Scheme\u20052, and a similar mixture of diastereoisomers, can also be formed by direct addition of a slight excess of H3B\u2009\u22c5\u2009PPhHBH2\u2009\u22c5\u2009PPhH22PCH2CH2PPh2)(\u03b76\u2010C6H5F)][BArF4]8\u2010toluene. ESI\u2010MS analysism/z=747.2 with the correct isotopologue pattern. Irrespective of the method of synthesis, accompanying 2 are unidentified minor hydride signals observed between \u221215 to \u221216\u2005ppm, that become dominant as catalysis reaches completion. These are currently unidentified,3B\u2009\u22c5\u2009PPhH2 they are not deactivation products. Complex 2, at least at the early stages of catalysis, is thus identified as a likely resting state.In situ catalyst speciation studies , using the distinctive P\u2212H resonances between 5.5 and 3.5\u2005ppm, reveals a first order decay of H3B\u2009\u22c5\u2009PPhH2 \u00d710\u22124\u2005s\u22121), linear diboraphosphine H3B\u2009\u22c5\u2009PPhHBH2\u2009\u22c5\u2009PPhH2 as an intermediate and the growth in of oligomeric/polymeric [H2BPPhH]n, Scheme\u20053B\u2009\u22c5\u2009PPhH2 restarts catalysis (kobs=1.5(1)\u00d710\u22124\u2005s\u22121). In a separate experiment (3\u2005mol% 1) using H3B\u2009\u22c5\u2009PPhHBH2\u2009\u22c5\u2009PPhH2 (0.14\u2005M) as the precursor resulted in its clean first order consumption (kobs=1.0(1)\u00d710\u22124\u2005s\u22121) to form [H2BPPhH]n . Combined these observations on the resting state and the temporal evolution of substrates and products lead us to propose a simplified mechanism, Scheme\u2005(i)\u2005the P/B dehydrocoupling of \u2212PPhH2 and \u2212BH3 end groups at the metal center to form 2 (R\u2032=H), or a close analog thereof, i.e. R\u2032=[PPhHBH2]nPPhH\u2009\u22c5\u2009BH3; (ii)\u2005a reversible chain transfer between bound and free phosphine\u2013boranes; (iii)\u2005a turnover limiting step that involves substitution of the newly coupled oligomer with another phosphine\u2013borane (monomer/oligomer/polymer), that leads to the observed first\u2010order decay of monomers H3B\u2009\u22c5\u2009PPhH2 or H3B\u2009\u22c5\u2009PPhHBH2\u2009\u22c5\u2009PPhH2; (iv)\u2005an overall step\u2010growth like mechanism that supports the observation of H3B\u2009\u22c5\u2009PPhHBH2\u2009\u22c5\u2009PPhH2 as a persistent intermediate.Following reaction progress by t 100\u2009\u00b0C \u2005mol% 1, t 100\u2009\u00b0C \u2005mol% 1, Mn of isolated polymer using higher loadings, but incomplete conversions with higher dispersities at lower loadings using the same reaction time clearly demonstrates step growth propagation (Scheme\u2005Mn=77\u2009000\u2005g\u2009mol\u22121) is only formed after 5\u2005days reaction time. In contrast, for a chain growth process, high molecular weight polymer would be expected to be observed at low conversions of monomer.n, Mn(MALS)=7\u2009000\u2005g\u2009mol\u22121, with H3B\u2009\u22c5\u2009P(n\u2010hexyl)H2 monomer (1.25\u2005M in toluene), catalyst 1 (3\u2005mol%), and heating for 66\u2005hrs at 100\u2009\u00b0C resulted in a mixture of products by 31P{1H} NMR spectroscopy, including homopolymer [H2BP(n\u2010hexyl)H]n and related short\u2010chain oligomers. Using the noted solubility difference between Ph\u2010 and n\u2010hexyl substituted polyphosphinoboranes, extraction into hexanes selectivity removed polymers/oligomers containing only n\u2010hexyl groups. Importantly the hexane insoluble portion displayed both phenyl and hexyl resonances in the corresponding 1H and 31P spectra, an initial indication that a block copolymer had been formed, Scheme\u200531P NMR spectrum a signal at \u03b4 \u221249.5 [J(PH)=350\u2005Hz] is assigned to [H2BPPhH]n segments, while a cluster of signals centered at \u221263.4 is assigned to [H2BP(n\u2010hexyl)H]n, interestingly in which the atacticity can now be observed,1H NMR spectrum in the P\u2212H region indicates a similar ratio of Ph to n\u2010hexyl units. Additional smaller signals are observed in the 31P NMR spectrum at \u03b4 \u221255.8 as an apparent triplet, that simplifies to a singlet on decoupling 1H, and \u03b4 \u221248 . These could reflect connecting units between block segments, P(n\u2010hexyl)H2 end groups110\u2010b\u2010[H2BP(n\u2010hexyl)H]11, BCP1. A small amount of low molecular weight oligomer (\u2248500\u2005g\u2009mol\u22121) is also observed in the GPC spectrum. As pre\u2010formed [H2BPPhH]n contained no such species, we assign this to short oligomers of [H2BP(n\u2010hexyl)H]n.With this calibration in hand, taking pre\u2010formed n and [H2BP(n\u2010hexyl)H]n have very different diffusion coefficients (1.5(2) and 3.9(8) \u00d710\u221210\u2005m2\u2009s\u22121, respectively, Figure S42). However, BCP1 also has a different diffusion coefficient, that is the same for both phenyl and hexyl regions of the 1H NMR spectrum (1.99(3) and 1.95(5) \u00d710\u221210\u2005m2\u2009s\u22121), indicating both segments are in the same macromolecule. A two\u2010component model is needed to best fit the data in the alkyl region, with a minor, faster diffusing, component required (9.6(2)\u00d710\u221210\u2005m2\u2009s\u22121). This is fully consistent with the shorter, oligomeric, material observed by GPC. Dynamic Light Scattering experiments place the apparent hydrodynamic radius of BCP1 as 2.7\u00b12.0\u2005nm in CHCl3 , a reasonable fit with that from DOSY (2.1\u00b10.2\u2005nm), and supports the formation of non\u2010aggregated polymer chains in CHCl3 solution.BCP1, the ability to reversibly bind and activate different terminus P\u2212H/B\u2212H groups is no doubt important to the formation of a block copolymer. As it is well established that P\u2212H activation occurs more quickly with PPhH2 than with P(n\u2010hexyl)H2 H2 or equivalent short chain oligomers.While we currently cannot comment on the precise order of events to form r Scheme\u2005, we suggs Scheme\u2005C, followBCP1 self\u2010assembled micellar nanostructures should be formed in solution. Molecularly dissolved BCP unimers were observed in pure THF and CHCl3, or in an equal volume mixture of THF and hexane by dynamic light scattering (DLS) . When the volume fraction of hexane was increased to 37.5\u2009%:62.5\u2009% v/v THF:hexane, to enhance the selectivity for the \u2212[H2BP(n\u2010hexyl)H]11 segment of BCP1, DLS provided evidence for the formation of self\u2010assembled aggregates with Rh=132\u00b112\u2005nm .\u2005Decreasing the solvent polarity further to 25\u2009%:75\u2009% THF:hexane resulted in larger aggregates being formed . The solvent composition and DLS data indicate that micelles are formed with a core of the more polar \u2212[H2BPPhH]110 block and a corona of the less polar \u2212[H2BP(n\u2010hexyl)H]11 segment. The core:corona block ratio ca. 10\u2009:\u20091 classifies these systems as crew\u2010cut micelles and non\u2010spherical morphologies, rather than star\u2010like micelles, would be anticipated based on packing\u2010parameter considerations.BCP1 in 37.5\u2009%:62.5\u2009% v/v THF:hexane solution revealed the formation of rod\u2010like micelles should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "In the crystal structure of the title compound, the packing is driven by C\u2014H\u22efF, N\u2014H\u22efO and C\u2014H\u22ef\u03c0 contacts. Hirshfeld surface analysis showed that the largest contribution to the surface contacts arise from F\u22efH/H\u22efF inter\u00adactions. 11H10F4N2O2, the conformation about the N\u2014C\u2014C\u2014O bond is gauche [torsion angle = 61.84\u2005(13)\u00b0]. In the crystal, N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into [010] chains, which are cross-linked by C\u2014H\u22efF and C\u2014H\u22ef\u03c0 contacts. Hirshfeld surface analysis was conducted to aid in the visualization of these various influences on the packing. This analysis showed that the largest contribution to the surface contacts arises from F\u22efH/H\u22efF inter\u00adactions (35.6%), followed by O\u22efH/H\u22efO (17.8%) and H\u22efH (12.7%).In the title compound, C The amine nitro\u00adgen atom (N2) and ester oxygen atom (O1) are gauche to one another, with N2\u2014C6\u2014C7\u2014O1 = 61.84\u2005(13)\u00b0. The C10 methyl group is oriented in such a fashion as to enable a weak C\u2014H\u22ef\u03c0 inter\u00adaction with the aromatic ring of an adjacent mol\u00adecule linker unit between the amine nitro\u00adgen atom and the ester, and F3 as acceptor. One of these inter\u00adactions is intra\u00admolecular (C6\u2014H6A\u22efF3) with the other being inter\u00admolecular (C7\u2014H7A\u22efF3). A hydrogen-bonding inter\u00adaction occurs between the secondary amine and the carbonyl oxygen atom (N2\u2014H1N2\u22efO2). Finally, a weak C\u2014H\u22ef\u03c0 inter\u00adaction is observed between H7B and the pyridine ring \u03c0 system.The main directional inter\u00adactions in the crystal structure of the title compound are of the type C\u2014H\u22efF, N\u2014H\u22efO and C\u2014H\u22ef\u03c0 Table\u00a01. The N\u2014HCrystalExplorer17.5 to 1.38 (blue) a.u.; the pale-red spots symbolize short contacts and negative dnorm values on the corresponding surface plots shown in Fig.\u00a02Hirshfeld surface analysis was used to investigate the presence of hydrogen bonds and inter\u00admolecular inter\u00adactions in the crystal structure. The Hirshfeld surface analysis < 1.15\u2005\u00c5. The H\u22efH and C\u22efH/H\u22efC contacts have the second and third largest contributions, at 17.8 and 12.7%, respectively. Smaller spikes on the shoulder of the fingerprint plot, appearing at 1.15\u2005\u00c5 < (id + ed) < 1.160\u2005\u00c5, correspond to the N\u22efH/H\u22efN contacts.The largest contribution to the overall crystal packing is from F\u22efH/H\u22efF inter\u00adactions (35.6%) Table\u00a02. However4.et al., 2016ConQuest amino]\u00adethan-1-ol was synthesized using a known method and used without further purification : \u03b4 6.01 , 5.61 , 4.90 , 4.38 , 3.86 , 1.93 \u2013, 3H); 19F NMR : \u03b4 \u221293.6 , \u2212163.4 ; 13C NMR : \u03b4 167.8 (C=O), 135.8 (CH2=C(CH3)-), 126.4 [CH2=C(CH3)\u2013], 63.7 (\u2013OCH2CH2NH\u2013), 44.1 (\u2013OCH2CH2NH\u2013), 18.3 [CH2=C(CH3)\u2013].A 500\u2005ml round-bottom flask equipped with an addition funnel was charged with 2-[(perfluoro\u00adpyridin-4-yl)amino]\u00adethan-1-ol , tri\u00admethyl\u00adamine and diethyl ether (300\u2005ml). The solution was stirred under nitro\u00adgen at 273\u2013278\u2005K for 15 minutes. Next, a solution of methacrylol chloride in ether (10\u2005ml) was added dropwise to the round-bottom flask using an addition funnel. The solution was allowed to gradually warm to room temperature and was stirred for 96\u2005h under nitro\u00adgen. Precipitated salts were removed by vacuum filtration and the filtrate was concentrated under reduced pressure. Crystals of the title compound in the form of colorless needles were obtained by recrystallization from a solution in warm (\u223c328\u2005K) hexa\u00adnes : m.p. 335\u2013336\u2005K; 6.Uiso(H) = 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023004334/hb8066sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023004334/hb8066Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023004334/hb8066Isup3.cmlSupporting information file. DOI: 2263932CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Details of correction: correct footnotes to tables\u00a01, 2 and 3; please correct each oneExisting text: in the footnotes of tables\u00a01,2 and 3:\u00b5g Vit D31 \u00b5g = 40 Corrected text should read:IU Vit D31 \u00b5g = 40"} +{"text": "The crystal structure of the bromo\u00adbenzene\u00adsulfonamide derivative of the type 2 diabetes drug metformin is presented. 10H14BrN5O2S, is the bromo\u00adbenzene\u00adsulfonamide derivative of the type 2 diabetes drug metformin. The asymmetric unit contains two mol\u00adecules with almost identical conformations but a different orientation of the bromo\u00adphenyl moiety. Both mol\u00adecules exhibit intra\u00admolecular N\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds. The mol\u00adecular packing features chain formation in the a-axis direction by alternating N\u2014H\u22efN and N\u2014H\u22efO inter\u00adactions. In addition, ring motifs consisting of four mol\u00adecules and \u03c0\u2013\u03c0 inter\u00adactions between the phenyl rings contribute to the three-dimensional architecture. A Hirshfeld surface analysis shows that the largest contributions to surface contacts arise from contacts in which H atoms are involved.The title compound, C Metformin works by decreasing gluconeogenesis in the liver, increasing insulin sensitivity and preventing insulin resistance in the asymmetric unit and N30\u22efN34 (mol\u00adecule B) are shared by the two nitro\u00adgen atoms with an occupancy of 0.85\u2005(4) at atoms N15 and N34, and 0.15\u2005(4) at atoms N11 and N30. The dihedral angles between the phenyl ring and the best plane through the N-containing moiety (N11\u2013C19 in A and N30\u2013C38 in B) are 87.12\u2005(12) and 96.05\u2005(12)\u00b0 in A and B, respectively. Next to the intra\u00admolecular hydrogen bonds N15\u2014H15B\u22efN11 and N34\u2014H34B\u22efN30, a short inter\u00adaction is present between atoms H16B and O9 in A, and H35B and O9 in B inter\u00adaction, while mol\u00adecule B forms an N34\u2014H34A\u22efO10 hydrogen-bond inter\u00adaction (Table\u00a01D11(2); Etter & MacDonald, 1990a-axis direction is formed by subsequent N16\u22efN32 and N34\u22efO10 inter\u00adactions distance is 3.686\u2005(3)\u2005\u00c5 and the slippage is 0.650\u2005\u00c5, while for mol\u00adecule B the Cg2\u22efCg2 distance is 4.1086\u2005(3)\u2005\u00c5 and the slippage is 1.936\u2005\u00c5 .Further dimer formation is obtained through \u03c0\u2013\u03c0 stacking between the phenyl rings Fig.\u00a06. For molCrystal Explorer21.3 , H16A\u22efN32 [2.54\u2005(5)\u2005\u00c5] and Br\u22efBr [3.4165\u2005(10)\u2005\u00c5] inter\u00adactions present in the crystal packing. The bright-red spots in Fig.\u00a08A and H35B refer to the N16\u2014H16A\u22efN32, N34\u2014H34A\u22efO10 and N35\u2014H35B\u22efO9 hydrogen bonds, while the additional faint-red spots illustrate weaker H15A\u22efN30 [2.70\u2005(5)\u2005\u00c5] and C14\u22efH23 (2.66\u2005\u00c5) inter\u00adactions present in the crystal packing. The relative distributions from the different inter\u00adatomic contacts to the Hirshfeld surfaces are presented in Table\u00a02A and B, respectively).A Hirshfeld surface analysis was performed, and two-dimensional fingerprint plots were created with 4.et al., 2016a resulted in 17 hits [DEXBUF and DEXBUF01 . For the title compound, this torsion angle is \u2212177.5\u2005(4) and \u2212171.8\u2005(4)\u00b0 in A and B, respectively.A search of the Cambridge Structural Database . The mixture was stirred for 30\u2005min at room temperature. After the reaction was complete, water was removed under reduced pressure and the residue was dissolved in cold anhydrous methanol. The sodium chloride was filtered off and the filtrate was evaporated under reduced pressure to obtain basic metformin.Metformin hydro\u00adchloride was dissolved in 12Cl2 = 1:10) to obtain the title compound as a colourless solid.The basic metformin and 3-bromo\u00adbenzene\u00adsulfonyl chloride were dissolved in 6\u2005mL of anhydrous di\u00adchloro\u00admethane and stirred under a nitro\u00adgen atmosphere for 3\u2005h at room temperature. The solvent was removed on a rotary evaporator and the residue was purified by column chromatography and sodium gliclazide were dissolved in 5\u2005mL of acetone and stirred overnight at room temperature. The solvent was removed under reduced pressure and a light-yellow solid was obtained, which was expected to be the sulfonyl\u00adurea salt of the title compound.Cuboid-shaped colourless crystals were grown in an NMR tube by slow evaporation over two weeks using deuterated chloro\u00adform as solvent. However, the grown crystals consist of the title compound and not of its sulfonyl\u00adurea salt.1H NMR \u03b4 8.03 , 7.90\u20137.76 , 7.64\u20137.57 , 7.56\u20137.22 , 7.06 , 5.19 , 2.99 . 13C NMR \u03b4 160.29 (s), 158.59 (s), 145.65 (s), 134.41 (s), 130.20 (s), 129.13 (s), 124.70 (s), 122.54 (s), 37.00 (s).NMR spectra of the title compound were recorded on a 400\u2005MHz NMR spectrometer: 6.Uiso(H) values assigned as 1.2Ueq or 1.5Ueq (methyl only) of the parent atoms, with C\u2014H distances of 0.93 (aromatic) and 0.96\u2005\u00c5 (meth\u00adyl). The hydrogen atoms bound to nitro\u00adgen were located in a difference-Fourier map and refined freely with Uiso(H) values assigned as 1.2Ueq of the parent atoms. The occupancy factors of hydrogen atoms H11 and H15B (mol\u00adecule A), and H30 and H34B (mol\u00adecule B) involved in intra\u00admolecular hydrogen bonds converged during refinement to 0.85\u2005(4) for H15B and H34B, and 0.15\u2005(4) for H11 and H30.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023002165/pk2683sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023002165/pk2683Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989023002165/pk2683sup3.pdfCSD survey results. DOI: Click here for additional data file.10.1107/S2056989023002165/pk2683Isup4.cmlSupporting information file. DOI: 2246792CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atom is coordinated by the S and N atoms of two N\u2032-[(Z)-(furan-2-yl)methyl\u00adidene]carbamohydrazono\u00adthioic acid ligands in a distorted square-planar geometry.In the title complex, the Ni 6H6N3OS)2]\u00b72CH3OH, the NiII atom is coordinated by the S and N atoms of two N\u2032-[(Z)-(furan-2-yl)methyl\u00adidene]carbamohydrazono\u00adthioic acid ligands in a distorted square-planar geometry. The two mutual ligands bound to NiII are also connected by C\u2014H\u22efS inter\u00adactions, while the H atoms of the NH2 group of the ligands form R44(8) motifs with the O atoms of the solvent ethyl alcohol mol\u00adecules. At the same time, the OH groups of the solvent ethyl alcohol mol\u00adecules form parallel layers to the (011) plane by the O\u2014H\u22efN inter\u00adactions with the ligand N atom that is not bonded to the NiII atom.. The layers are connected by van der Waals inter\u00adactions. A Hirshfeld surface analysis indicates that the most important contacts are H\u22efH (37.7%), C\u22efH/H\u22efC (14.6%), O\u22efH/H\u22efO (11.5%) and S\u22efH/H\u22efS (10.6%).In the title complex, [Ni(C The ligands assume a trans arrangement with respect to each other around the NiII ion, which lies on a crystallographic inversion centre at . The Ni\u2014S [2.1818\u2005(6)\u2005\u00c5] and Ni\u2014N [1.9055\u2005(17)\u2005\u00c5] bond lengths lie within the range of those found in related structures.Fig.\u00a013.II are also linked by C\u2014H\u22efS inter\u00adactions, while the H atoms of the NH2 group of the ligands form et al., 1995II atom is shown in Fig.\u00a05A Hirshfeld surface analysis was carried out using b; 37.7%) contacts to be the most common, followed by C\u22efH/H\u22efC , O\u22efH/H\u22efO and S\u22efH/H\u22efS contacts. The N\u22efH/H\u22efN (8.5%), O\u22efC/C\u22efO (4.9%), Ni\u22efH/H\u22efNi (3.2%), O\u22efN/N\u22efO (2.2%), N\u22efC/C\u22efN (1.9%), C\u22efC (1.8%), S\u22efC/C\u22efS (1.1%), S\u22efS (0.7%), O\u22efO (0.7%),S\u22efO/O\u22efS (0.5%) and Ni\u22efC/C\u22efNi (0.2%) contacts have little directional influence on the mol\u00adecular packing.The two-dimensional fingerprint plots show the H\u22efH Fig.\u00a06b; 37.7%\u22efC Fig.\u00a06c; 14.6%\u22efO Fig.\u00a06d; 11.5%\u22efS Fig.\u00a06e; 10.6%4.et al., 2016et al., 1987et al., 2009A search of the Cambridge Structural Database \u2005\u00c5 and Ni\u2014N(2) = 1.921\u2005(2)\u2005\u00c5. The coordination around Ni is trans planar with respect to the two S and two N atoms. The furan ring plane is at an angle of 3(1)\u00b0 to the coordination plane. In the crystal of NOQCUS, the coordination environment around the nickel(II) ion is totally planar, as the NiN2S2 chromophore lies on its least-squares calculated plane and the four angles formed by the metal centre with the four donor atoms add up to exactly 360\u00b0. The Ni\u2014N and Ni\u2014S distances are within the usual range. This plane forms a 18\u00b0 angle with the uncoordinated furan ring, which is also highly planar.In the crystal of FUTRAN, Ni 5.E)-2-(furan-2-yl\u00admethyl\u00adene)hydrazine-1-carbo\u00adthio\u00adamide were dissolved in 30\u2005mL of methanol then 13\u2005mg (0.05\u2005mmol) of Ni(OOCCH3)2\u00b74H2O were added. The reaction mixture was kept in air at room temperature for slow evaporation. After ca 2\u20133\u2005d, orange crystals, suitable for X-ray analysis, were formed.17\u2005mg (0.1\u2005mmol) of (14H20N6NiO4S2 (M = 459.17); C 36.61 ; H 4.35 (4.39); N 18.26 (18.30) %. IR (KBr): 3372 \u03bd(OH), 2965 and 2854 \u03bd(NH), 1643 \u03bd(C=N) cm\u22121.Yield 81%, soluble in DMSO, ethanol and di\u00admethyl\u00adformamide and insoluble in non-polar solvents. Elemental analysis: C6.Uiso(H) = 1.2 or 1.5Ueq(C). O- and N-bound H atoms were located in difference Fourier maps and refined with Uiso(H) = 1.2Ueq(N) and 1.5Ueq(O), with their positions fixed. Two reflections (001) and (010), affected by the beam stop, were omitted in the final cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023005182/jy2031sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989023005182/jy2031Isup2.hklStructure factors: contains datablock(s) I. DOI: 2269284CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In all three title crystals, the cations are linked by O\u2014H\u22efO and/or C\u2014H\u22efO hydrogen bonds. The three-dimensional packing is further consolidated by strong halogen\u2013hydrogen and weak van der Waals inter\u00adactions. N,N-di\u00admethyl\u00adacetamide\u20131-(dimethyl-\u03bb4-aza\u00adnyl\u00adidene)ethan-1-ol tribromide (1/1), C4H9NO\u00b7C4H10NO+\u00b7Br3\u2212 or [(C4H9NO)\u00b7(C4H10NO)](Br3), (I), N,N-di\u00admethyl\u00adacetamide\u20131-(dimethyl-\u03bb4-aza\u00adnyl\u00adidene)ethan-1-ol di\u00adbromido\u00adiodate (1/1), C4H9NO\u00b7C4H10NO+\u00b7Br2I\u2212 or [(C4H9NO)\u00b7(C4H10NO)](Br2I), (II), and N,N-di\u00admethyl\u00adacetamide\u20131-(dimethyl-\u03bb4-aza\u00adnyl\u00adidene)ethan-1-ol di\u00adchlorido\u00adiodate (1/1), C4H9NO\u00b7C4H10NO+\u00b7Cl2I\u2212 or [(C4H9NO)\u00b7(C4H10NO)]\u00b7(Cl2I), (III), all the anions are almost linear in geometry and all the cations, except for the methyl H atoms, are essentially planar. In the crystal structure of (I), the cations are linked by pairs of C\u2014H\u22efO hydrogen bonds, forming inversion dimers with an R22(8) ring motif. These dimers also exhibit O\u2014H\u22efO hydrogen bonding. Dimerized cation pairs and anions are arranged in columns along the a axis. In the crystal of (II), the cations are linked by pairs of O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming an R44(14) ring motif. These groups of cations and the anions form individual columns along the a axis and jointly reside in planes roughly parallel to (011). In the crystal of (III), cations and anions also form columns parallel to the a axis, resulting in layers parallel to the (020) plane. Furthermore, the crystal structures of (I), (II) and (III) are consolidated by strong halogen (Br and/or I and/or Cl)\u22efH and weak van der Waals inter\u00adactions. In addition to the structural evaluation, a Hirshfeld surface analysis was carried out.In the title compounds, The cations, except for their methyl H atoms, are essentially planar [r.m.s. deviation = 0.041\u2005(1)\u2005\u00c5 for O1]. For (II), the angles and distances of the anion are Br1\u2014I1\u2014Br2 = 177.942\u2005(5)\u00b0, I1\u2014Br1 = 2.7244\u2005(2)\u2005\u00c5 and I1\u2014Br2 = 2.68597\u2005(19)\u2005\u00c5. These values are in agreement with data reported in the literature , I1 resides in the centre of inversion symmetry [Cl1\u2014I1\u2014Cl1\u00a0= 180.0\u00b0], with distances of I1\u2014Cl1 = 2.53973\u2005(18)\u2005\u00c5. The cations, except for their methyl H atoms, are planar and all reside on mirror planes.In the title compounds Figs. 1 and 3 \u25b8,I), (II) and (III), the O\u2014C and N\u2014C bond distances of the cation all fall between single and double bond values, with C1\u2014N1 = 1.3134\u2005(17)\u2005\u00c5 and C1\u2014O1 = 1.2786\u2005(16)\u2005\u00c5 for (I), C1\u2014N1 = 1.3168\u2005(16)\u2005\u00c5, C5\u2014N2 = 1.3121\u2005(16)\u2005\u00c5, C1\u2014O1 = 1.2771\u2005(15)\u2005\u00c5 and C5\u2014O2 = 1.2794\u2005(15)\u2005\u00c5 for (II), and C1\u2014N1 = 1.3161\u2005(8)\u2005\u00c5 and C1\u2014O1 = 1.2750\u2005(8)\u2005\u00c5 for (III). The corresponding bond lengths of the three compounds are in good agreement with each other and with the literature.In (3.I), the cations are linked by pairs of C\u2014H\u22efO hydrogen bonds , forming inversion dimers with an et al., 1995x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a02). Dimerized cation pairs and anions are arranged in columns along the a axis , two cations are refined in the asymmetric unit. These cations are linked by pairs of O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming an a axis and reside in planes parallel to (011) , cations and anions are arranged in columns parallel to the a axis, forming layers parallel to the (020) plane (Table\u00a03I), (II) and (III) are consolidated by strong halogen (Br and/or I and/or Cl)\u22efH bonding inter\u00adactions, Coulombic attraction and weak van der Waals inter\u00adactions , (II) and (III) are 2.4224\u2005(15), 2.4278\u2005(14) and 2.4261\u2005(9)\u2005\u00c5, respectivly, and are thereby within the range (2.31\u20132.63\u2005\u00c5) found for short/strong classical hydrogen bonds , (II) and (III) were carried out and created with CrystalExplorer17.5 , (II) and (III) is given in Table\u00a04I), (II) and (III) are shown in Fig.\u00a010b)\u2013(d) and Table\u00a05I); 60.3% for (II); 88.9% for (III)], Br\u22efH/H\u22efBr , O\u22efH/H\u22efO [6.5% for (III)] and O\u22efH/H\u22efO and C\u22efH/H\u22efC [2.0% for (III)] contacts.The Hirshfeld surface analysis and the associated two-dimensional fingerprint plots over the cations of (I): space group, monoclinic P21/n, Z = 2; (II): space group, triclinic PZ = 2; (III): space group, monoclinic C2/m, Z = 2], may be the result of small deviations in the inter\u00adactions arising from the different crystal systems and packing, as well as from the variations in the anions of the compounds.The respective differences in the crystal structures of the three title compounds [aluminium(III)] bis\u00ad\u00adium hepta\u00adkis\u00ad(perchlorate) tetra\u00adchloro\u00adgold(III) , the Al3+ ion is surrounded by dma mol\u00adecules (dma = di\u00admethyl\u00adacetamide) in an octa\u00adhedral arrangement. The dma mol\u00adecules are essentially planar. Three Al\u2014O\u2014C\u2014N torsion angles [138.8\u2005(8)\u2013149.3\u2005(4)\u00b0] are found to deviate significantly from 180\u00b0. The centrosymmetric cation has the bridging H atom at the centre of inversion. The planar structure is essentially the same as those reported for [H(dma)2]+ cations; the O\u22efO distance [2.386\u2005(8)\u2005\u00c5] is within the range (2.31\u20132.63\u2005\u00c5) found for short hydrogen bonds , the structure consists of distinct [AuCl4]\u2212 anions and [H(dma)2]+ cations, with the gold and the bridging H atoms located at centres of symmetry. The hydrogen bond is \u2018symmetrical\u2019 as a result of crystallographic requirements. The O\u22efO distance is 2.430\u2005(16)\u2005\u00c5. Thermal motion analysis indicates that methyl groups attached to nitro\u00adgen have higher rotational amplitudes, resulting in short apparent C\u2014H bond lengths [average 0.96\u2005(4)\u2005\u00c5] compared with the methyl group attached to a carbonyl C atom which has an average C\u2014H bond length of 1.02\u2005(2)\u2005\u00c5.In the crystal of HDMAAU , two N,N-di\u00admethyl\u00adacetamide mol\u00adecules in the asymmetric unit are connected to each other by an O\u2014H\u22efO hydrogen bond, essentially sharing the central H atom. These mol\u00adecules and the Br\u2014Br\u2014Br groups are arranged in columns parallel to the a axis. The arrangement is consolidated in the crystal packing by van der Waals inter\u00adactions between these columns.In the crystal of SEGMOG , the unit-cell parameters and the arrangement of the mol\u00adecules are relatively similar to the older structure (SEGMOG), while the H atom bridging the the two acetamides was not refined.In the crystal of SEGMOG01 hydrogen halides as orange colored solids. Single crystals of bis\u00ad hydrogen halides were obtained by slow crystallization from methanol.To a solution of di\u00admethyl\u00adacetamide in 0.09\u2005mol of 38% hydro\u00adchloric or 40% hydro\u00adbromic acid under stirring and cooling in an ice\u2013water bath, 0.05\u2005mol iodine monochloride , iodine monobromide or bromine was added gradually. The mixture was stirred for 1\u2005h and the crystals were filtered off, dried and recrystallized from methanol to give the target bis\u00ad: 1664 (NCO). 1H NMR : \u03b4 12.51 , 3.28 , 3.19 , 2.45 ; 13C{1H} NMR : \u03b4 174.5, 39.7, 37.5, 19.9.Bright orange crystals (Rodygin 5.3.\u22121): 1606 (NCO). 1H NMR : \u03b4 10.72 , 3.28 , 3.19 , 2.46 ; 13C{1H} NMR : \u03b4 174.6, 39.6, 37.5, 20.1.Bright-orange crystals, yield 44% (10.2\u2005g), m.p. 343\u2013344\u2005K. IR (KBr), \u03bd (cm5.4.\u22121): 1611 (NCO). 1H NMR : \u03b4 9.98 , 3.25 , 3.17 , 2.41 ; 13C{1H} NMR : \u03b4 174.2, 39.4, 37.2, 19.8.Bright orange crystals, yield 75% (14\u2005g), m.p. 364\u2013365\u2005K. IR (KBr), \u03bd (cm6.I), (II) and (III), the C-bound H atoms were positioned geometrically, with C\u2014H = 0.98\u2005\u00c5 (for methyl H atoms), and constrained to ride on their parent atoms, with Uiso(H) = 1.5Ueq(C). The hy\u00addroxy H atoms were found in the difference Fourier maps and their coordinates were refined freely, with Uiso(H) = 1.5Ueq(O). In (I), the H atom of the OH group is located in a special position with an occupancy of 0.5 for the rrefined atom. In (II), the H atoms of the OH groups are disordered over two positions, with occupancies of 0.49 and 0.51. In (III), the H atom of the OH group was refined with an occupancy of 0.25 for its position close to an inversion centre in between the O atoms of two acetamides and simultaneously residing on a mirror plane.Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989023005509/yz2034sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989023005509/yz2034Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989023005509/yz2034IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989023005509/yz2034IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 2271693, 2271692, 2271691CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Thus, we highlight thesections where these changes apply.In a recent Letter,Abstract: The DO\u2013Dbond dissociation energy has been reportedto be 5.41 \u00b1 0.10 eV, while considering EA(O) in the originalmanuscript. However, EA(OD) should be used instead, thus leading toa bond dissociation energy of 5.28 \u00b1 0.20 eV.Page 5365,left column, second paragraph; \u201cFrom the appearance...\u201dis reorganized and reads as follows:Table S4 togetherwith the BDE, the enthalpy of reaction can be obtained from \u0394Hr(OH\u2013) = D(H\u2013OH) \u2013 EA(OH) = 3.34 eV. In the charge transfer process,if we add the potassium ionization energy, OH\u2013 isexpected at 7.68 eV. The reactions threshold was obtained assumingno excess energy (E#), yet momentum conservationof the dissociating partners may impact on the lighter fragment kineticenergy, thus shifting the energy to a higher value. We note a differenceof \u223c1.1 eV for the energy loss data, which is certainly plausiblegiven the kinetic-energy release distribution of H\u2013 in Figure S3. Thus, from the appearanceenergy (AE) in the H2O energy loss spectrum at \u0394E \u2248 8.8 eV, one can obtain the HO\u2013Hbond dissociation energy (BDE) by taking the potassium ionizationenergy and the data from Table S4,2 i.e., D(HO\u2013H) = AE(OH\u2013) \u2013 IE(K) + EA(OH) \u2013 E#, yielding therefore D(HO\u2013H)= 5.19 \u00b1 0.20 eV which is in good agreement with the values 5.15eV 3 and 5.17eV.4 Following the same approach for D2O, the energy loss spectrum shows a threshold feature at \u0394E \u2248 9.0 eV. We obtain for the first time the DO\u2013Dbond dissociation energy to be D(DO\u2013D) = 5.28\u00b1 0.20 eV. In D2O the DO\u2013D energy value isslightly higher than the O\u2013D bond dissociation energy (5.176eV5), which is in agreement with its analogueH2O.Taking the values in Page 5366, right column, last paragraph; \u201cElectronicstatespectroscopy...\u201d reads as follows:2O/D2O was thoroughly discussedfrom the experimental K+ energyloss spectra obtained, from which the DO\u2013D bond dissociationenergy has been obtained for the first time to be 5.28 \u00b1 0.20eV.The electronic statespectroscopy of HfHg\u00b0) in reactions 2a.1\u20132b.2and 4a.1\u20134b.2are actually enthalpies of reaction (\u0394Hr). Table S2 decimal places forO\u2013 and OH\u2013 from H2Ohave been corrected; they now read 12.08 \u00b1 0.20 and 7.80 \u00b10.20.Enthalpies of formation (\u0394"} +{"text": "However, the difficulty in expanding cytotoxic \u03b3\u03b4T cells to an adequate number has been a major limitation to the efficacy of treatment in most cases. We successfully re-generated \u03b3\u03b4T cells from \u03b3\u03b4T cell-derived human induced pluripotent stem cells (iPSCs). The iPSC-derived \u03b3\u03b4T cells (i\u03b3\u03b4Ts) killed several cancer types in a major histocompatibility complex (MHC)-unrestricted manner. Single-cell RNA sequencing (scRNA-seq) revealed that the i\u03b3\u03b4Ts were identical to a minor subset of PB\u03b3\u03b4Ts. Compared with a major subset of PB\u03b3\u03b4Ts, the i\u03b3\u03b4Ts showed a distinctive gene expression pattern: lower CD2, CD5, and antigen-presenting genes; higher CD7, KIT, and natural killer (NK) cell markers. The i\u03b3\u03b4Ts expressed granzyme B and perforin but not interferon gamma (IFN\u03b3). Our data provide a new source for \u03b3\u03b4T cell-based immunotherapy without quantitative limitation.For a long time, \u2022We successfully re-generated \u03b3\u03b4T cells from \u03b3\u03b4T cell-derived human iPSCs\u2022The re-generated \u03b3\u03b4T cells showed cytotoxicity against cancer cell lines\u2022Distinctive signatures of the re-generated \u03b3\u03b4T cells were identified ex vivo-expanded peripheral-blood-derived \u03b3\u03b4T cells. Cytotoxicity of i\u03b3\u03b4Ts against several cancer cell lines was exhibited in an MHC-unrestricted manner. The novel cell resource will advance off-the-shelf \u03b3\u03b4T cell-based immunotherapy.Murai et\u00a0al. successfully re-generated \u03b3\u03b4T cells from \u03b3\u03b4T cell-derived human iPS cells. Molecular signatures of the re-generated \u03b3\u03b4T cells (i\u03b3\u03b4Ts) were identical to those of a minor subset of Therefoical use . Howeverical use . Furtherex\u00a0vivo expansion rate of PBMC-derived \u03b3\u03b4T cells varies widely from donor individual to individual; thus, ex\u00a0vivo-expanded autologous PB\u03b3\u03b4T cell therapy cannot be applied to all patients; (2) patients have to wait for the expansion of autologous cells but not for off-the shelf allogenic cells; and (3) an autologous approach would be associated with high costs. iPSCs have infinite proliferation ability: they show logarithmic growth for at least 100\u00a0days and \u223c1,028-fold expansion during 100\u00a0days (Induced pluripotent stem cell (iPSC) technology may be able to overcome these limitations and enable us to realize off-the-self allogenic \u03b3\u03b4T cell-based therapy, which has several advantages over autologous \u03b3\u03b4T cells therapy: (1) the 2\u00a0weeks) . To gene2\u00a0weeks) , and re-2\u00a0weeks) .bona fide lymphocyte, including \u03b3\u03b4T cells, and should be categorized as cells resulting from an aberrant characteristic of lymphocytes derived from iPSCs, or abnormal cells. Previous studies have demonstrated that such abnormal cells can be derived from pre-rearranged TCR-carrying pluripotent stem cells or hematopoietic progenitors, and such iPSCs reportedly gave rise to abnormal T\u00a0cells expressing TCR, an NK cell marker NK1.1, and CD8\u03b1\u03b1 exhibited cytotoxicity against several cancer cell lines in an MHC-unrestricted manner. We identified distinctive molecular signatures of i\u03b3\u03b4Ts and clarified that the i\u03b3\u03b4Ts were identical to a minor subset of in\u00a0vitro embryoid body (EB)-mediated differentiation experiment showed that they could differentiate into three germ layers stimulated with HMBPP for 1\u00a0week being composed of both \u03b3\u03b4TCR-positive and \u03b1\u03b2TCR-positive cells, CD3-positive i\u03b3\u03b4Ts contained no \u03b1\u03b2TCR-positive cells A. Genomiin i\u03b3\u03b4Ts B. Furthein i\u03b3\u03b4Ts , whereasin i\u03b3\u03b4Ts ).Figure\u00a0These data indicated that we successfully re-generated monoclonal \u03b39\u03b42 T\u00a0cells via \u03b3\u03b4T-iPSCs.A key advantage of \u03b3\u03b4T cells for cancer immunotherapy is that one type of \u03b3\u03b4T cell is applicable for various types of cancer in a human leukocyte antigen (HLA)-unrestricted manner . We therFirst, we labeled Jurkat cells with carboxyfluorescein diacetate succinimidyl ester (CFSE) as target cells, co-cultured with the i\u03b3\u03b4Ts as effector cells at an effector (E):target (T)\u00a0ratio of 2:1 and stained these cells with 7-aminoactinomycin-D (7-AAD) to identify dead cells, followed by flow cytometry (FCM). The no-effector condition showed that only approximately 5% of Jurkat cells were dead B, althouTo assess the cytotoxicity of i\u03b3\u03b4Ts toward solid tumor cells, we observed the co-culture of 62B3 \u03b3\u03b4T-iPSC line-derived i\u03b3\u03b4Ts with GFP-Huh-7 cells by time-lapse imaging. After 12\u00a0h of co-culture, the areas of GFP-Huh-7 cells decreased to 28.7%, 30.1%, and 64.5% of those in no-effector control culture in three independent experiments C and 3D.This video displays the continuous imaging of i\u03b3\u03b4Ts co-cultured with GFP-labeled Huh-7 cells at an E:T ratio of 2:1 from 0 to 6 h. The scale bar (upper left) indicates 100\u00a0\u03bcm.Next, to confirm the cytotoxicity of purified-i\u03b3\u03b4Ts, we co-cultured CD3-MACS-purified i\u03b3\u03b4T cells and tumor cells and quantified the tumor toxicity at an E:T ratio of 2:1 at 12\u00a0h using xCELLigence. Against Huh-7 cells, the purified i\u03b3\u03b4T and PB\u03b3\u03b4T cells showed no significant difference in cytotoxicity G. MoreovThese data demonstrated that the i\u03b3\u03b4Ts have cytotoxicity for at least four different types of cancer cells in an HLA-unrestricted manner. Moreover, we were able to catch i\u03b3\u03b4Ts coming into contact with GFP-Huh-7 cells and peeling off as time went on .We next performed several experiments to obtain insight into the mode of action of the i\u03b3\u03b4Ts. The co-culture of i\u03b3\u03b4Ts and Jurkat cells at various E:T ratios showed the dose-dependent cytotoxic effects of i\u03b3\u03b4Ts A and S3ENext, we investigated the mechanism by which i\u03b3\u03b4Ts exhibit cytotoxicity. Both blocking antibodies for \u03b3\u03b4TCR and NKG2D reduced the cytotoxicity of the purified i\u03b3\u03b4Ts,\u00a0suggesting that the i\u03b3\u03b4Ts recognize tumor cells by both\u00a0\u03b3\u03b4TCR and NKG2D C. PerforNotably, no i\u03b3\u03b4Ts were positive for IFN\u03b3. In contrast, most granzyme B-positive cells in PB\u03b3\u03b4T cells were positive for IFN\u03b3 D. This fWe compared the cell surface markers of i\u03b3\u03b4Ts with PB\u03b3\u03b4T cells by FCM E. Both dIn general, T\u00a0cells are divided into four subsets of naive or memory phenotypes corresponding to the CD45RA and CD27 expression patterns . Despitein\u00a0vitro for 7\u00a0days and CD3(+) \u03b3\u03b4TCR(+) cells were sorted. (iii) i\u03b3\u03b4Ts; differentiated cells from \u03b3\u03b4T-iPSC clone 62B3 (according to the protocol shown in \u03b3\u03b4T cells have been reported to have various subtypes and it wTRDC(+), CD3E(+), CD4(\u2212), CD8A(+), and CD8B(\u2212) (TRDC(+), CD3E(+), CD4(\u2212) CD8A(\u2212), and CD8B(\u2212) B and S4A CD8B(\u2212) B and S4Ased SELL B. Cells onocytes C. Cells and CD3E B, but diclusters D. We catThe t-SNE distribution of each sample and the fraction of clusters in each sample are shown in To examine the expression of major immune-related genes in each \u03b3\u03b4T subset, we created dot plots for T\u00a0cell-differentiation marker genes , cytokinCTSW, FCER1G, KLRC3, CD244, NKG7, as well as the cytotoxic marker perforin coding gene [PRF1]). Cells in \u03b3\u03b4T subset 2, the dominant population of in\u00a0vitro-expanded PB\u03b3\u03b4Ts stimulated with HMBPP, expressed immune checkpoint inhibitory receptors and IFN\u03b3 and IFN\u03b3-inducing genes . Cells in \u03b3\u03b4T subset 1, the main population of i\u03b3\u03b4Ts, were enriched for NK cell-related genes E. The doB, IRF4) were exp\u03b4T cells , was notectively C. The ex 1 and 3 A and 6C. 1 and 3 A and 6C.In order to investigate the reproducibility of the i\u03b3\u03b4T data, the expression of these marker genes in scRNA-seq data of i\u03b3\u03b4Ts prepared three times independently was shown by a heatmap. The similar expression patterns indicated that the gene expression of i\u03b3\u03b4Ts was reproducible E.Taken together, we successfully re-generated MHC-unrestricted cytotoxic \u03b3\u03b4T cells from iPSCs and clarified the\u00a0distinctive molecular signatures of iPSC-derived \u03b3\u03b4T cells.in\u00a0vivo.In the present study, we successfully re-generated CD3(+)\u03b3\u03b4TCR(+) cells from \u03b3\u03b4T cell-derived iPSCs. Although there have been reports of the re-generation of \u03b1\u03b2T cells from \u03b1\u03b2T cell-derived iPSCs , it was ex\u00a0vivo-expanded PB\u03b3\u03b4Ts, indicating that the cells resembling the major population of i\u03b3\u03b4Ts exist in adult PBMCs in nature but are not expandable with HMBPP stimulation, at least under the culture condition used in this study. Previously there have been many studies concerning the classification of human peripheral \u03b3\u03b4T cells. For example, the functions of \u03b3\u03b4T cells were reportedly separated into five subsets: IFN-\u03b3-producing, antigen-presenting, follicular B helper, regulatory \u03b3\u03b4T, and IL-17-producing cells , and CD5(\u2212) \u03b3\u03b4T cells were reported to be more cytotoxic than CD5(+) \u03b3\u03b4T cells . In addiNK cell-related markers were expressed in i\u03b3\u03b4Ts. A subset of authentic \u03b3\u03b4T cells reportedly expressed NK cell-related genes and recognize target cells by a similar mechanism to NK cells. In addition, it is reported that mimetic-\u03b3\u03b4 NKT cells, which expressed low T\u00a0cell-related genes and high NK cell-related genes, were induced from iPSCs . The shaWe herein demonstrated that our i\u03b3\u03b4Ts were completely negative for \u03b1\u03b2TCR and that they killed tumor cells in an MHC-independent manner. The negative expression of \u03b1\u03b2TCR may reduce the risk of graft-versus-host disease . For thiSeveral limitations of the present study should be addressed in our future studies. First, the induction efficiency of i\u03b3\u03b4Ts was not satisfactory, and we have not clarified what CD3(\u2212) cells existing after i\u03b3\u03b4Ts induction were. Second, it should be evaluated whether or not the i\u03b3\u03b4Ts attack non-cancer cells of KIR-ligand mismatch recipients. Third, the i\u03b3\u03b4T induction protocol established in this study used xenogenic serum and feeder cells, which are difficult for clinical applications. We are currently trying to generate i\u03b3\u03b4Ts under feeder-free and serum-free conditions (data not shown). Our technologies will advance off-the-shelf \u03b3\u03b4T cell-based immune therapy.This study did not generate new unique reagents.3 cells and cultured in StemFit medium .Seven days before induction, human \u03b3\u03b4T-iPSCs were seeded onto a six-well plate at a density of 2.0\u00a0\u00d7\u00a010On day 0, the medium was completely replaced by StemFit medium supplemented with 4\u00a0\u03bcM CHIR99021 , 80\u00a0ng/mL BMP4 , and 80\u00a0ng/mL vascular endothelial growth factor (VEGF) . On day 2, the medium was replaced by Essential 6 medium supplemented with 2\u00a0\u03bcM SB431542 , 50\u00a0ng/mL bFGF , 50\u00a0ng/mL SCF , and\u00a080\u00a0ng/mL VEGF. On day 4, the medium was replaced by StemPRO-34 SFM supplemented with 2\u00a0mM L-glutamine, 50\u00a0ng/mL IL-3 , 50\u00a0ng/mL IL-6 , 50\u00a0ng/mL FLT3L , 50\u00a0ng/mL SCF, 20\u00a0ng/mL VEGF, and 10 IU/mL EPO .On days 6 and 8, the medium was replaced with StemPRO-34 SFM supplemented with 2\u00a0mM L-glutamine, 50\u00a0ng/mL IL-6, 50\u00a0ng/mL SCF, and 10 IU/mL EPO.On day 10, hematopoietic cells were transferred into wells co-cultured with feeder cells. Floating cells and supernatant were collected in the tube. Adhesive cells were dissociated with Accutase , and incubated at 37\u00b0C for 10\u00a0min. Supernatant was returned to the well, pipetted, and filtered using a 35-\u03bcm cell strainer. After centrifugation at 1,200\u00a0rpm for 4\u00a0min, cells were suspended in OP9 medium supplemented with 10\u00a0ng/mL SCF, 10\u00a0ng/mL TPO , 5\u00a0ng/mL IL-7 , 5\u00a0ng/mL FLT3L, and 100\u00a0\u03bcg/mL L-ascorbic acid . Cells were reseeded to the same well and incubated at 37\u00b0C for 30\u00a0min. Without pipetting, supernatant and floating cells were transferred into a new well confluent with pre-seeded OP9/N-DLL1 cells.On day 12, half of the medium was changed and cells were transferred into new wells with fresh OP9/N-DLL1 cells by vigorous pipetting. Then, half of the medium was changed every other day and cells were transferred onto fresh OP9/N-DLL1 cells every 6\u00a0days.On day 30, we collected cells with Accutase, similarly to day 10. Cells were suspended with RPMI1640 medium supplemented with 10% FBS, 1\u00a0nM HMBPP , 100 IU/mL IL-2 , and 10\u00a0\u03bcM 2-mercaptoethanol and seeded onto new plates in a feeder-free condition. Half of the medium was changed every other day. After more than a week of stimulation, cytotoxicity was analyzed.The day before the targeted scRNA-seq analysis, CD3(+)\u03b3\u03b4TCR(+) cells were sorted on a BD FACS Aria III from PB\u03b3\u03b4Ts and i\u03b3\u03b4Ts that were stimulated with HMBPP for the indicated days as described above. To infer the origin of the sample, all cells were labeled with multiplex sample tags. Single-cell capture and cDNA library preparation were performed using a BD Rhapsody Single-Cell Analysis System with a BD Human Single-Cell Multiplexing Kit and BD Human Immune Response Targeted Panel for Human , which contains 399 primer pairs, targeting 397 different genes, according to the manufacturer\u2019s recommendations. The concentration, size, and integrity of the resulting PCR products were assessed using a Qubit High-Sensitivity dsDNA Kit.Sequencing was performed using an Illumina HiSeq X in Macrogen . Fastq files were uploaded to the Seven Bridges Genomics online platform. The obtained counts were adjusted by distribution-based error correction (DBEC), an error correction algorithm developed by BD Biosciences. DBEC data were then loaded into Seurat (version 4.0.4.). Cells were then clustered using a resolution of 0.03 and visualized by t-SNE. The Seurat functions FeaturePlot, DotPlot, DoHeatmap, and Vlnplot were used to visualize the gene expression with feature plot, dot plot, heatmap, and violin plot, respectively. Markers for a specific cluster against all remaining cells were found by using the Seurat function FindAllMarkers.GSE194072.ScRNA-seq data have been deposited in GEO under accession number Data are expressed as the mean\u00a0\u00b1 SD. Differences between two groups were analyzed using a paired t test. Statistical analyses were performed using Microsoft Excel 2013 and EZR. p values of <0.05 were considered statistically significant.Conceptualization, T.A; methodology, N.M; software, T.A. and M.K.-A; validation, M.K.-A.; formal analysis M.K.-A.; investigation, N.M.; resources, N.M.; writing\u00a0\u2013 original draft, N.M.; writing\u00a0\u2013 review\u00a0& editing, T.A., M.K.-A.; supervision, T.A. and H.T.; project administration, T.A."} +{"text": "The quinoxaline unit in the title mol\u00adecule is slightly puckered [dihedral angle between the rings = 2.07\u2005(12)\u00b0] while the whole mol\u00adecule adopts an L-shaped conformation. The packing in the crystal is governed by C\u2014H\u22efO hydrogen bonds and slipped \u03c0-stacking inter\u00adactions. 18H16N4O5, is slightly puckered [dihedral angle between the rings = 2.07\u2005(12)\u00b0] while the whole mol\u00adecule adopts an L-shaped conformation. Intra\u00admolecular hydrogen bonding determines the orientation of the substituted phenyl ring and the amide nitro\u00adgen atom is almost planar. The packing in the crystal is governed by C\u2014H\u22efO hydrogen bonds and slipped \u03c0-stacking inter\u00adactions.The quinoxaline unit in the title mol\u00adecule, C The oris Table\u00a01. H3A mayn Table\u00a01. The quic-axis direction \u2005\u00c5, dihedral angle = 2.07\u2005(10)\u00b0, slippage alternates between 1.40 and 1.28\u2005\u00c5 along the stack]. The \u03c0-stacking is reinforced by C8=O1\u22efCg1 inter\u00adactions, where Cg1 is the centroid of the C1/C6/N1/C7/C8/N2 ring: O1\u22efCg1 = 3.3333\u2005(16)\u2005\u00c5, C8\u22efCg1 = 3.689\u2005(2)\u2005\u00c5, C8=O1\u22efCg1 = 96.89\u2005(2)\u00b0. The stacks are linked by C10\u2014H10A\u22efO2 and C16\u2014H16\u22efO4 hydrogen bonds -one was dissolved in 25\u2005ml of di\u00admethyl\u00adformamide, then 1.53\u2005g (6.24\u2005mmol) of 2-chloro-N-(4-meth\u00adoxy-2-nitro\u00adphen\u00adyl)acetamide were added followed by 1.0\u2005g (7.5\u2005mmol) of potassium bicarbonate, and a spatula tip of BTBA [2-benzyl\u00adsulfanyl-5-(tri\u00adfluoro\u00admeth\u00adyl)benzoic acid] was used for the phase-transfer catalysis. The reaction was stirred for 2\u2005h under reflux at 80\u00b0C. When the starting reagents had completely reacted, 500\u2005ml of distilled water were added and a few minutes later the product precipitated. This was filtered, dried and recrystallized from hot ethanol solution to yield light-yellow plate-like crystals of the title compound.A mass of 1.00\u2005g (6.24\u2005mmol) of 3-methyl\u00adquinoxalin-2 global, I. DOI: 10.1107/S2414314623001918/hb4426Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623001918/hb4426Isup3.cmlSupporting information file. DOI: 2245645CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In this DFT mechanistic work, a novel substrate\u2010dependent catalytic mechanism is disclosed involving cooperative Ca\u2212H\u2212Ca bridges for H2 isotope exchange, competitive Ca\u2212H\u2212Ca bridges and terminal Ca\u2212H bonds for anti\u2010Markovnikov addition of unactivated 1\u2010alkenes, and terminal Ca\u2212H bonds for Markovnikov addition of conjugation\u2010activated styrene. THF\u2010coordination plays a key role in favoring the anti\u2010Markovnikov addition while strong cation\u2010\u03c0 interactions direct the Markovnikov addition to terminal Ca\u2212H bonds.Recently, it was shown that the double Ca\u2212H\u2212Ca bridged calcium hydride cation dimer complex [LCaH Calcium hydride cation dimer catalysis: Dispersion\u2010corrected DFT calculations reveal that the same double Ca\u2212H\u2212Ca bridged calcium hydride cation dimer catalyst [LCaH2CaL]2+\u22c5THF shows a novel substrate\u2010dependent catalytic activity, involving cooperative Ca\u2212H\u2212Ca bridges for efficient H2 isotope exchange, terminal Ca\u2212H bond for the Markovnikov addition of conjugation\u2010activated styrene, and competitive Ca\u2212H\u2212Ca bridges and terminal Ca\u2212H bonds for the anti\u2010Markovnikov addition of unactivated 1\u2010alkenes. In THF solution, the weakly (or non\u2010) coordinating counter\u2010anions BAr4\u2212 should be solvated into separated ions, and thus not considered further. Consistent with experiment,2+1 is indeed \u22122.8\u2005kcal/mol exergonic, with a decisive stabilizing dispersion contribution of \u22127.6\u2005kcal/mol otherwise the coordination should be 4.8\u2005kcal/mol endergonic and thus thermodynamically unstable. For comparison, the THF coordination to the cation monomer +1\u2009m is only \u22121.5\u2005kcal/mol exergonic with a smaller dispersion contribution of \u22124.2\u2005kcal/mol. The formation of double Ca\u2212H\u2212Ca bridged 2+1 from two +1\u2009m monomers is \u221218.4\u2005kcal/mol exergonic again with a sizable dispersion contribution of \u22126.2\u2005kcal/mol. The THF coordination to 2+1\u22c5THF smoothly leads to two +1\u2009m\u22c5THF cation monomers that is 15.3\u2005kcal/mol endergonic, indicating a rapid dimer\u2010to\u2010monomer equilibrium even at room temperature (see Supporting Information Table\u2005S1).To gain deep mechanistic insight into the cation dimer 22Ca(BDI)]2 is about 7.7\u2005kcal/mol higher in free energy than two (THF)(BDI)CaH monomers in benzene solution,2Ca(BDI)] is 40.4\u2005kcal/mol lower in enthalpy than two (BDI)CaH monomers in gas\u2010phase,In two related mechanistic studies on 1\u2010alkene hydrogenation reactions, the THF\u2010coordinated [(THF)(BDI)CaH]2+1\u22c5THF, the anti\u2010Markovnikov addition of unactivated 1\u2010alkene CH2=CHR (R=3\u2010cyclohexenyl) to a Ca\u2212H\u2212Ca bridge of 2+1 after THF\u2010elimination is 3.6\u2005kcal/mol endergonic over a sizable free energy barrier of 23.8\u2005kcal/mol (via transition state 2+TS1) to selectively form the dinuclear complex 2+A [LCaHR'CaL]2+ containing both a Ca\u2212H\u2212Ca and a Ca\u2212R\u2019\u2212Ca bridge. The alternative Markovnikov addition with the hydride added to the terminal alkene carbon is kinetically 7.6\u2005kcal/mol less favorable . The subsequent H2 addition over the Ca\u2212R\u2019\u2212Ca bridge of 2+A is highly exergonic and kinetically 5.8\u2005kcal/mol more favorable (via 2+TS2) to form the hydrogenated product CH3CH2R along with regenerated 2+1\u22c5THF after THF coordination. Such a catalytic 1\u2010alkene hydrogenation is thus \u221223.7\u2005kcal/mol exergonic over a sizable barrier of 23.8\u2005kcal/mol involving anti\u2010Markovnikov alkene addition to cooperative Ca\u2212H\u2212Ca bridges.As shown in Figure\u2005+1\u2009m\u22c5THF that is 7.8\u2005kcal/mol higher in free energy than 2+1\u22c5THF, the anti\u2010Markovnikov addition of unactivated CH2=CHR to the terminal Ca\u2212H bond is still 4.0\u2005kcal/mol endergonic but over nearly the same free energy barrier of 23.8\u2005kcal/mol (via +mTS1) to selectively form the calcium alkyl complex +mA [LCaHR\u2019\u22c5THF]+, with the alternative Markovnikov addition being kinetically 6.0\u2005kcal/mol less favorable . The subsequent hydrogenolysis of the Ca\u2212C bond of +mA with H2 is highly exergonic and kinetically 2.9\u2005kcal/mol more favorable (via +mTS2) to form the hydrogenated product CH3CH2R along with regenerated +1\u2009m\u22c5THF. When cyclohexene is used as an internal alkene substrate in +1\u2009m\u22c5THF catalyzed hydrogenation, a 3.3\u2005kcal/mol higher barrier is found for the anti\u2010Markovnikov addition to the terminal Ca\u2212H bond, consistent with the experimentally observed selective hydrogenation of terminal alkene.On the other hand, via the THF\u2010coordinated cation monomer +1\u2009m is kinetically 0.7\u2005kcal/mol less reactive than that of +1\u2009m\u22c5THF towards CH2=CHR addition, mainly due to a less Lewis\u2010basic hydride (Mulliken atomic charges: \u22120.16 e in +1\u2009m vs. \u22120.53 e in +1\u2009m\u22c5THF). Together with the positive THF affinity of +1\u2009m, this leads to a 2.1\u2005kcal/mol higher barrier of 25.9\u2005kcal/mol for CH2=CHR addition without the coordinating THF. Interestingly, despite nearly the same free energy barriers at 298\u2005K for the competitive mechanisms via 2+TS1 and +mTS1, our DFT calculations show that the former via the solvent\u2010free dimer 2+1 becomes kinetically 1.4\u2005kcal/mol more favorable than the latter via the THF\u2010coordinated +1\u2009m\u22c5THF upon heating at 60\u2009\u00b0C (333\u2005K) in experiment,2+1\u22c5THF to reach the solvent\u2010free 2+TS1. In contrary to the usual intuition that the dimer\u2010to\u2010monomer conversion should be favored upon heating due to favorable entropy effects, our DFT calculations show that the equilibrium of 2+1\u22c5THF+THF\u2192+1\u2009m\u22c5THF++1\u2009m\u22c5THF in solution is hardly affected by such temperature change due to negligible entropy difference with an unchanged number of molecules on both sides (see Supporting Information Table\u2005S1).Interestingly, after THF elimination, the terminal Ca\u2212H bond of solvent\u2010free 2 isotope exchange may occur via one of two cooperative Ca\u2212H\u2212Ca bridges of the solvent\u2010free cation dimer 2+1 over a low barrier of 16.9\u2005kcal/mol (via 2+TS3) after THF\u2010elimination from stable 2+1\u22c5THF catalyst. Interestingly, three exchanging hydrogen atoms are placed evenly between two calcium ions and perpendicular to the remaining Ca\u2212H\u2212Ca bridge, suggesting potentially strong cooperative effects. With the coordinating THF within 2+1\u22c5THF, similar H2 isotope exchange (via 2+TS3a) becomes kinetically 4.5\u2005kcal/mol less favorable. On the other hand, facile H2 isotope exchange may also occur via terminal Ca\u2212H bonds that are more reactive but higher in free energy than Ca\u2212H\u2212Ca bridges. Indeed, the THF\u2010coordinated cation monomer +1\u2009m\u22c5THF is intrinsically 3.9\u2005kcal/mol (via +mTS3) more reactive than 2+1 for H2 isotope exchange but 7.8\u2005kcal/mol higher in free energy, eventually leading to a 3.9\u2005kcal/mol higher barrier. Interestingly, the solvent\u2010free cation monomer +1\u2009m with a less Lewis\u2010basic hydride turns out to be intrinsically 3.9\u2005kcal/mol less reactive than +1\u2009m\u22c5THF in mediating H2 isotope exchange, which is further disfavored by its positive THF affinity of 1.4\u2005kcal/mol. It is thus clear that very efficient H2 isotope exchange observed at room temperature2+1 over a low barrier of 16.9\u2005kcal/mol (via 2+TS3) rather than recently proposed terminal Ca\u2212H bonds.As seen in Figure\u20052+1\u22c5THF, the Markovnikov addition of styrene (CH2=CHPh) as a typical \u03c0\u2010conjugation\u2010activated 1\u2010alkene to a cooperative Ca\u2212H\u2212Ca bridge of 2+1 after THF\u2010elimination is 0.2\u2005kcal/mol endergonic over a sizable barrier of 24.3\u2005kcal/mol (via 2+TS4), selectively leading to the dinuclear calcium\u2010complex 2+B [LCaHR\u201cCaL]2+ containing both a Ca\u2212H\u2212Ca and a Ca\u2212R\u201d\u2212Ca Ph) bridge. Note that the hydride is now added to the terminal rather than the inner alkene carbon as directed by strong Ca+\u2026Ph cation\u2010\u03c0 interactions; the anti\u2010Markovnikov addition encounters a 2.7\u2005kcal/mol higher barrier and thus is kinetically disfavored. The subsequent hydrogenolysis of the Ca\u2212R\u201d\u2212Ca bridge of 2+B with H2 is highly exergonic and kinetically 7.7\u2005kcal/mol more favorable (via 2+TS5) to form the hydrogenated product CH3CH2Ph along with regenerated catalyst 2+1\u22c5THF after THF\u2010coordination. Such dimeric catalytic mechanism is \u221222.3\u2005kcal/mol exergonic over a sizable barrier of 24.3\u2005kcal/mol involving Markovnikov 1\u2010alkene addition to cooperative Ca\u2212H\u2212Ca bridges.As shown in Figure\u20052+1\u22c5THF, the Markovnikov styrene addition to the terminal Ca\u2212H bond of the solvent\u2010free cation monomer +1\u2009m (via +mTS4) is now kinetically very efficient and again directed by strong Ca+\u2026Ph ion\u2010\u03c0 interaction, which is now \u22127.7\u2005kcal/mol exergonic over a moderate barrier of 20.6\u2005kcal/mol to selectively form the calcium benzyl complex +mB LCaR\u201c+. The alternative anti\u2010Markovnikov styrene addition is 6.4\u2005kcal/mol less favorable and is kinetically highly disfavored. The subsequent H2 hydrogenolysis of the Ca\u2212C bond of +mB is \u221214.6\u2005kcal/mol exergonic over a moderate barrier of 19.0\u2005kcal/mol (via +mTS5) to form the hydrogenated product CH3CH2Ph along with regenerated catalyst 2+1\u22c5THF after +1\u2009m dimerization and THF\u2010coordination, which is kinetically 1.6\u2005kcal/mol more favorable than the preceding Markovnikov styrene addition. It is thus clear that the catalytic styrene hydrogenation via the terminal Ca\u2212H bond of the solvent\u2010free cation monomer +1\u2009m encounters only a moderate barrier of 20.6\u2005kcal/mol, which is kinetically 3.7\u2005kcal/mol more favorable than that via cooperative Ca\u2212H\u2212Ca bridges of the solvent\u2010free cation dimer 2+1 and reasonably accounts for the efficient styrene hydrogenation observed at room temperature.Interestingly, starting from the stable complex 2 isotope exchange reactions with the same calcium hydride cation dimer catalyst 2+1\u22c5THF. It is shown that cooperative Ca\u2212H\u2212Ca bridges of solvent\u2010free dimer 2+1 are favored for H2 isotope exchange, competitive Ca\u2212H\u2212Ca bridges of 2+1 and the terminal Ca\u2212H bond of cation monomer +1\u22c5THF are involved in the anti\u2010Markovnikov addition of unactivated 1\u2010alkenes, while the terminal Ca\u2212H bond of solvent\u2010free monomer +1 is preferred for the Markovnikov addition of conjugation\u2010activated styrene as directed by strong cation\u2010\u03c0 interactions. The novel mechanism can reasonably explain the known experimental observations and may be a useful guide for rational catalyst design.Extensive dispersion\u2010corrected DFT calculations disclose a novel substrate\u2010dependent catalytic mechanism of 1\u2010alkene hydrogenation and Hsolv=3.18\u2005\u00c5). The density\u2010fitting RI\u2212J approachAll DFT calculations were performed with the TURBOMOLE 7.4 suite of programs.More accurate solvation free energies in THF solution were computed with the COSMO\u2010RS modelThe authors declare no conflict of interest.1As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "P21/c symmetry with four mol\u00adecules in the unit cell. The imidazole ring is rotated 80.7\u2005(1)\u00b0 relative to the phenyl ring. Inter\u00admolecular stabilization primarily results from close contacts between the N-atom at the 3-position on the imidazole ring and the C\u2014H bond at the 4-position on the neighboring DippIm.At 106\u2005(2)\u2005K, the title mol\u00adecule has monoclinic 15H20N2 or DippIm, is reported. At 106\u2005(2)\u2005K, the mol\u00adecule has monoclinic P21/c symmetry with four mol\u00adecules in the unit cell. The imidazole ring is rotated 80.7\u2005(1)\u00b0 relative to the phenyl ring. Inter\u00admolecular stabilization primarily results from close contacts between the N atom at the 3-position on the imidazole ring and the C\u2014H bond at the 4-position on the neighboring DippIm, with ar\u00adyl\u2013aryl distances outside of the accepted distance of 5\u2005\u00c5 for \u03c0-stacking.The crystal structure of the title compound, C They are precursors in many synthetic processes and find use in pharmaceuticals and agrochemicals to create anti\u00adfungal agents and fungicides in H2O, followed by addition of H2NCH2CH(OEt2), and acidic workup with HCl and HNO3 ligands, which are a common ligand class for organometallic chemistry and catalysis or potassium bis\u00ad(tri\u00admethyl\u00adsil\u00adyl)amide (KHMDS) to form the free carbene ligands reported by our group is reported.Few aryl\u00adimidazoles have been structurally characterized, with 1--12.DippIm crystallizes as depicted in Fig.\u00a01MesIm of 106.44\u2005(16), 105.65\u2005(17), 110.89\u2005(18), 104.54\u2005(17), and 112.48\u2005(17)\u00b0, respectively and 1.3759\u2005(16)\u2005\u00c5, respectively \u2005\u00c5, C1\u2014N2 = 1.316\u2005(3)\u2005\u00c5, C2\u2014C3 = 1.356\u2005(3)\u2005\u00c5, N1\u2014C3 = 1.384\u2005(2)\u2005\u00c5, and N2\u2014C2 = 1.382\u2005(3)\u2005\u00c5 \u2005\u00c5 between N1\u2014C1 and a longer bond of 1.3153\u2005(16)\u2005\u00c5 between C1\u2014N2, likely due to steric effects of the aryl group. The backbone imidazole C2\u2014C3 bond distance of 1.3578\u2005(16)\u2005\u00c5 is consistent with a C3.Mercury program\u2019s centroid algorithm -1H-imidazole. A SciFinder search of colorless crystals. The product was characterized with 1H NMR and the results were consistent with reported literature values I. DOI: 10.1107/S2056989023009179/oi2001Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023009179/oi2001Isup3.cmlSupporting information file. DOI: 2302037CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Correction: Neural Dev 18, 1 (2023)10.1186/s13064-022-00169-1The authors would like to correct errors and update two figures in the original publication of the article .1. Page 2, \u201cAll mouse lines were backcrossed for at least 6 generations\u201d corrected to \u201cAll mouse lines were backcrossed for at least 4 generations\u201d.2. Page 4, \u201c0.1\u00a0M boric acid pH 8\u201d corrected to \u201c0.1\u00a0M sodium borate pH 8\u201d.3. Page 4, after \u201cRFP goat polyclonal Rockland Immunochemicals 200-101-379 1:500\u201d and before \u201cRFP rabbit polyclonal Rockland Immunochemicals 600-401-379 1:500\u201d, addition of \u201cRFP guinea pig polyclonal Frontier Institute MSFR105900 1:500\u201d.4. Page 4, \u201cTo our knowledge, this mouse line was not previously characterized.\u201d corrected to \u201cTo our knowledge, this mouse line was not previously characterized in detail.\u201d.2\u201d corrected to \u201cEGFP\u2009+\u2009KI-67\u2009+\u2009ASCL1\u2009+\u2009cells, 3373\u2009\u00b1\u2009532 cells per mm2\u201d.5. Page 6, \u201cKI-67\u2009+\u2009ASCL1\u2009+\u2009EGFP\u2009+\u2009cells, 3373\u2009\u00b1\u2009532 cells per mm6. Page 6, \u201cConsistent with the notion that the EGFP\u2009+\u2009KI-67- ASCL1- cells are quiescent B1 type stem cells\u201d corrected to \u201cConsistent with the notion that at least some of the EGFP\u2009+\u2009KI-67- ASCL1- cells are quiescent B1 type stem cells\u201d.7. Page 12, \u201call of the RFP\u2009+\u2009postnatal radial glial cells had cell body under the ventricular wall and contacted the ventricle with an apical extension .\u201d corrected to \u201calmost all of the RFP\u2009+\u2009postnatal radial glial cells had cell body under the ventricular wall and contacted the ventricle with an apical extension .\u201d8. Figure\u00a09. Figure"} +{"text": "This novel combination of a stereochemical probe and isotopic labeling in a silicon\u2010stereogenic compound opens new analytic possibilities to study stereochemical courses of reactions with the aid of chiral silanols mechanistically.A silicon\u2010stereogenic aminosilanol was isolated in excellent diastereomeric ratio and the absolute configuration was determined. The silanol is configurative and condensation stable in solution and shows stereoselective transformations with a clean stereospecific pathway in follow\u2010up reactions, which leads to the isolation of a silicon\u2010stereogenic zinc complex and siloxane compounds. Investigations with Silanols are important industrial building blocks for the functionalization of surfaces. In this research, one silicon\u2010stereogenic silanol was selectively synthesized and analyzed. By using an innovative combination of this stereochemical probe and isotope labeling, a configurationally stable chiral silanol was studied, providing new insights into mechanistic studies. Silanols are a central class of compounds in silicon chemistry and are widely used in synthesis,4, 5 cHowever, the preparation of Si\u2010chiral compounds has proven challenging in the past due to limited access to suitable prochiral or chiral precursors.24, 25Despite these initial findings on the limited configurational stability of silanols in solution, this aspect was not investigated further. Questions arise here, whose answers are essential in order to maximize the potential applications of chiral silanols. How stable is the configuration of chiral silanols in solution? Which subsequent reactions can be realized without loss of stereo information to make further functional components accessible?1 like siloxides and siloxanes.Thus, it can be used as an efficient mechanistic probe whose potential can be extended by 1 was synthesized by the acid\u2010catalyzed hydrolysis of the corresponding methoxysilane . Silanol 1 crystallizes in the space group P21. The quality of the crystal structure allowed the localization and free refinement of all protons. Two silanol molecules were observed in the asymmetric unit. These pack with the molecules generated by symmetry operations along the a\u2010axis to form long, chain\u2010like structures and are connected by intermolecular hydrogen bonds between the hydroxyl group of silanol 1 and the amine function of the next silanol.After hydrolysis, silanol 1 would be configurationally stable. Moreover, silanol 1 does not show condensation to the siloxane to provide the conditions for clean conversion reactions. For this purpose, the silanol was dissolved in different solvents and 1H\u2010 and 29Si\u2005NMR spectra of the compound were recorded over a period of several days and THF (8\u2005days) also showed high stability of silanol 1 in the time periods studied.In the following, conditions were sought in which silanol 1 under the conditions was confirmed, it was converted in first transformation reactions. The focus was on the selective synthesis of chiral siloxanes and siloxides. In this case, access to Si\u2010chiral representatives is of great interest, especially for the development of new materials. In addition, the metallated siloxides often show outstanding catalytic activity and access to Si\u2010stereogenic ligands could open the development of new components for chiral catalysis. to be able to detect the minor diastereomer as well. To ensure constant experimental conditions, the solution was stored in the refrigerator (T=4\u2009\u00b0C) for the duration of the experiment.For this purpose, a THF solution of silanol r Scheme\u2005. The comSSi/RSi) were determined after derivatization to siloxane 5. In parallel, the most intense oxygenated molecule fragments were used to determine the respective proportions of 18O in the corresponding diastereomers by GC/EI\u2010MS. The evaluation of the experiments shows an unexpectedly long stability of the configuration at the silicon center, which at first glance suggests no exchange with water. However, at the same time, the incorporation of large amounts of 18O into the diastereomers of silanol 1 is observed, making the labeled species the major components in solution at the end of the experimental period. Since this exchange occurs without loss of stereochemical information, it can be concluded that the reaction proceeds cleanly with retention of the configuration.During the experiment, samples were taken at regular intervals and the corresponding ratios of the diastereomers \u2010isomer exchanges with water faster than the (RSi)\u2010isomer of 1. It is possible that the amino substituent influences the exchange process through the formation of hydrogen bonds.These show that a stable configuration is not necessarily associated with frozen reactivity, since initial experiments of chiral silanol 1 that proceed in THF with retention of the configuration, thus ensuring long term stability of the configuration in the aqueous environment. The combination of a stereochemical probe and isotopic labeling experiments opens the possibility for large\u2010scale mechanistic studies of the central processes of silanols, which could provide access to new synthesis opportunities for silanol\u2010based materials.In summary, we were able to demonstrate access to a Si\u2010chiral silanol that is stable to configurational changes and to condensation reactions in solution, which can be transformed in clean stereochemical transformations to siloxanes and siloxides. In addition, an isotope labeling experiment revealed hidden exchange processes of water with silanol General remarks: All reactions with oxygen\u2010 and moisture\u2010sensitive compounds were performed under an atmosphere of argon with Schlenk\u2010techniques in dried solvents. The solvents were purified and dried by distillation over sodium and storage under argon atmosphere. Commercially available reagents were used without further purification except for phenylmethyldimethoxysilane, which was distilled and stored over molsieve (4\u2005\u00c5) in argon atmosphere. The NMR spectra were measured on a Bruker Avance III HD NanoBay \u2013 400\u2005MHz, 500\u2005MHz Bruker Avance NEO, 600\u2005MHz Bruker Avance III HD and 500\u2005MHz Agilent Technologies DD2 spectrometer at 25\u2009\u00b0C. Chemical shifts (\u03b4 in ppm) are referred to tetramethylsilane (TMS), with the deuterium signal of the solvent serving as internal lock and the residual solvent signal as additional reference 1H\u2005NMR \u03b4 (C6D5H)=7.16\u2005ppm). Signals were assigned with the help of HSQC experiments. For the multiplicities following abbreviations were used: s=singlet, br=broad signal, d=doublet, q=quartet. In diastereomeric associations, diastereomeric signals were assigned to the major diastereomer (Dmaj) and the minor diastereomer (Dmin) if distinguishable. For spectra measured in non\u2010deutered solvents, a capillary with C6D6 was added and served as an internal standard. 13C\u2005NMR Spectra were measured with broadband decoupling and referred to the signal of the solvent [{1H}13C\u2005NMR \u03b4 (C6D6)=128.39\u2005ppm]. {1H}29Si\u2005NMR spectra were as inverted gated with TMS as external standard. Single crystal X\u2010ray diffraction for compounds 1 and 6 were conducted on a Bruker D8 Venture four\u2010circle diffractometer by Bruker AXS GmbH using a PHOTON II CPAD detector by Bruker AXS GmbH. X\u2010ray radiation was generated by a microfocus source I\u03bcS Mo by Incoatec GmbH with HELIOS mirror optics and a single\u2010hole collimator by Bruker AXS GmbH. For the data collection, the programs APEX 4 Suite (v.2020.10\u20100) with the integrated programs SAINT (integration) and SADABS (absorption correction) by Bruker AXS GmbH were used.MiTeGen were used for mounting. For mass spectrometric evaluation a GC/EI\u2010MS system with a nominal\u2010resolution ISQ mass spectrometer coupled to a Thermo Trace GC Ultra oven and a direct probe controller from Thermo Fischer Scientific was used. The capillary column used was an OPTIMA from Macherey\u2010Nagel. Helium was used as the carrier gas. For high resolution mass spectrometry data a LTQ\u2010Orbitrap (Linear Trap Quadrupole Orbitrap) from Thermo Fischer Scientific coupled to a Shimadzu HPLC consisting of a CBM\u201020A communication module, a SPD\u2010M30A UV detector, a CTO\u201020AC column oven, a SIL\u201030AC autosampler, a LC\u201020ADXR pump system, and a DGU\u201020A5R degasser unit were used. Elemental analyses were performed with the elemental analyzer vario MICRO cube from the company Elementar and the weighing of the substance quantities was done with the microbalance Cubis MSE3.6P from the company Sartorius. Optical rotations were measured with an A. Kr\u00fcss Optical polarimeter operating on the sodium D\u2010line (589\u2005nm) using a quartz glass cuvette (1\u2005mL) and are reported as: [\u03b1]DT. The melting point for compound 1 was obtained using a B\u00fcchni M\u2010560 melting point apparatus and the resulting value is non\u2010corrected.Using Olex2, the structures were solved with the ShelXT structure solution program using Intrinsic Phasing and refined with the ShelXL refinement package using Least Squares minimization. MicroGrippers from Compound (S)\u20102: (S)\u20107 was suspended portion wise in 95\u2009% formic acid under ice cooling and 37\u2009% formaldehyde solution was subsequently added portion wise. The reaction solution was then stirred for 6\u2005h under reflux and then for 12\u2005h at room temperature. Subsequently, the reaction solution was adjusted to a pH of 13 by adding 2\u2005M sodium hydroxide solution. The aqueous phase was extracted with Et2O (3\u00d750\u2005ml) and the combined organic phases were dried over Na2SO4. After removal of the solvent, the residue was purified by \u201cKugelrohr\u201d\u2010distillation . The desired product (S)\u20102 was obtained as a colorless oil . 1H\u2005NMR : \u03b4=1.23 , 2.08 , 3.07 , 7.08\u20137.12 , 7.18\u20137.21 , 7.31\u20137.33 . 1H}13C\u2005NMR{ : \u03b4 =21.1 , 43.7 , 66.6 , 127.4 , 128.1 , 128.9 , 145.9 . GC/EI\u2010MS: tR=6.04\u2005min [80\u2009\u00b0C (1\u2005min)\u20137\u2009\u00b0C/min\u2013170\u2009\u00b0C (2.5\u2005min)\u201050\u2009\u00b0C/min\u2013250\u2009\u00b0C (1\u2005min)]; m/z (%): 149 (24.5) [M+], 134 (100) [(M\u2212Me)+], 105 (47) [(M\u2212NMe2)+], 91 (23), 77 (36) [(Ph)+], 72 (79) [(M\u2212Ph)+]. Specific rotation: [\u03b1]D20=\u221265.02\u00b0\u2005mL\u2009g\u22121\u2009dm\u22121 .Compound \u20103: (S)\u20102 was dissolved in Et2O (80\u2005ml). The solution was cooled to 0\u2009\u00b0C and t\u2010Butyllithium was dropped into solution. The yellowish solution was stirred for 1.5\u2005h at 0\u2009\u00b0C. Phenylmethyldimethoxysilane was then added to the solution and the solution was slowly thawed to room temperature with stirring and stirred for 24\u2005h in total. The solids were separated and the solvent was removed in vacuum. After purification by \u201cKugelrohr\u201d\u2010distillation at reduced pressure , the product \u20103 was isolated as a colorless oil . 1H\u2005NMR : \u03b4=Dmaj 0.63, Dmin 0.66 , Dmin 1.04, Dmaj 1.08 , Dmaj 1.76, Dmin 1.81 , Dmin 3.34, Dmaj 3.36 , Dmin 3.52, Dmaj 3.57 , 7.16\u20137.18 , 7.21\u20137.33 , 7.54\u20137.59 , 7.63 , 8.04\u20138.11 . 1H}13C\u2005NMR{ : \u03b4=Dmaj \u22123.0, Dmin \u22121.9 , Dmin 17.4, Dmaj 18.9 , Dmin 42.5, Dmaj 42.8 , Dmaj 50.8, Dmin 50.9 , Dmin 63.7, Dmaj 64.2 , 126.7 , 128.3 , 129.8 , 130.9 , 134.6 , 136.9 , 138.0 , 138.6 , 153.3 . 1H}29Si\u2005NMR{ : \u03b4=Dmin \u22126.3, Dmaj \u22125.4 (1Si). GC/EI\u2010MS: tR=5.99\u2005min [80\u2009\u00b0C (1\u2005min)\u201310\u2009\u00b0C\u2005min\u22121\u20131\u2013250\u2009\u00b0C (5.5\u2005min)]; m/z (%): 299 (1) [(M+)], 284 (17) [(M\u2212CH3)+], 268 (3) [(M\u2212OCH3)+], 252 (9) [(M\u2212OCH3\u2212CH3)+], 223 (7) [(M\u2212Ph)+], 206 (100) [(M\u2212Ph\u2212CH3\u2212H)+]. Elemental analysis: Calc.: C: 72.19\u2009%, H: 8.41\u2009%, N: 4.68\u2009%. Found: C: 72.0\u2009%, H: 8.4\u2009%, N: 4.4\u2009%.Compound \u20101: \u20103 was dissolved in Et2O (80\u2005ml). 2\u2005M HCl was added to the solution and the emulsion was intensively stirred for 2\u2005h. After that the reaction mixture was then cooled to 0\u2009\u00b0C and adjusted to pH\u200513 with KOH. The phases were separated and the aqueous phase was extracted with Et2O (3\u00d715\u2005mL). The combined organic phases were dried with MgSO4 and the volume of the solution was concentrated to 30\u2005ml. After storage at \u221230\u2009\u00b0C for 12\u2005h, the product \u20101 was obtained in the form of colorless crystals . 1H\u2005NMR : \u03b4=Dmaj 0.63, Dmin 0.66 , Dmin 1.04, Dmaj 1.08 , Dmaj 1.76, Dmin 1.81 , Dmin 3.34, Dmaj 3.36 , Dmin 3.52, Dmaj 3.57 , 7.16\u20137.18 , 7.21\u20137.33 , 7.54\u20137.59 , 7.63 , 8.04\u20138.11 . 1H}13C\u2005NMR{ : \u03b4=Dmaj \u22123.0, Dmin \u22121.9 , Dmin 17.4, Dmaj 18.9 , Dmin 42.5, Dmaj 42.8 , Dmaj 50.8, Dmin 50.9 , Dmin 63.7, Dmaj 64.2 , 126.7 , 128.3 , 129.8 , 130.9 , 134.6 , 136.9 , 138.0 , 138.6 , 153.3 . 1H}29Si\u2005NMR{ : \u03b4 =Dmin \u22126.3, Dmaj \u22125.4 (1Si). GC/EI\u2010MS: tR=5.99\u2005min [80\u2009\u00b0C (1\u2005min)\u201310\u2009\u00b0C\u2005min\u22121\u20101\u2013250\u2009\u00b0C (5.5\u2005min)]; m/z (%): 299 (1) [(M+)], 284 (17) [(M\u2212CH3)+], 268 (3) [(M\u2212OCH3)+], 252 (9) [(M\u2212OCH3\u2212CH3)+], 223 (7) [(M\u2212Ph)+], 206 (100) [(M\u2212Ph\u2212CH3\u2212H)+].Elemental analysis: Calc.: C: 72.19\u2009%, H: 8.41\u2009%, N: 4.68\u2009%. Found: C: 72.0\u2009%, H: 8.4\u2009%, N: 4.4\u2009%.Compound (S)\u20105: \u20101 was dissolved in Et2O (3.0\u2005mL). Hexamethyldisilazane was then added to the reaction solution. The solution was then stirred for 20\u2005h. The solvent and volatiles were removed under vacuum and the siloxane (S)\u20105 was obtained as a colorless oil . 1H\u2005NMR : \u03b4=0.13 , 0.69 , 1.14 , 1.86 , 3.50 , 7.15\u20137.17 , 7.22 , 7.32 , 7.54\u20137.57 , 7.74 , 7.95 . 1H}13C\u2005NMR{ : \u03b4=1.0 , 2.5 , 21.0 , 43.4 , 64.8 , 126.7 , 127.0 , 128.3 , 128.5 129.8 , 130.9 , 134.3 , 136.0 , 136.2 , 140.6 , 153.3 . 1H}29Si\u2005NMR{ : \u03b4 =Dmin\u201014.8, Dmaj \u221213.6 , Dmin \u22128.4, Dmaj \u22128.9 . GC/EI\u2010MStR=6.00\u2005min [80\u2009\u00b0C (1\u2005min)\u201320\u2009\u00b0C/min\u2013290\u2009\u00b0C (2\u2005min)]; m/z: 342 (10) [(M\u2212Me)+], 297 (14) {[M\u2212H3CHN(CH3)2]+}, 264 (100) [(M\u2212Ph\u2212Me)+], 209 (21) {[PhMeSiOSi(CH3)3]+}, 72 (41) {[Si(CH3)3]+}, 59 (1) {[CH2N(CH3)2]+}. m/z: 344 (10) [(18M\u2212Me)+], 299 (14) {[18M\u2212H3CHN(CH3)2]+}, 266 (100) [(18M\u2212Ph\u2212Me)+], 211 (4) {[PhMeSi18OSi(CH3)3]+}, 72 (40) {[Si(CH3)3]+}, 59 (1) {[CH2N(CH3)2]+}. LC/HR\u2010MS: tR=6.23\u2005min [0.3\u2005mL/min H2O/MeCN (+ 0.1\u2009% MeCOOH): 95\u2009:\u20095 (2\u2005min)\u201380\u2009:\u200920 \u20130\u2009:\u2009100 \u201395\u2009:\u20095 ]; 358.20225\u2005m/z {C20H32ONSi2 [M+H]+, (\u0394=1.552\u2005ppm)} 360.20618\u2005m/z {C20H3218ONSi2 [M+H]+, (\u0394=0.666\u2005ppm)} Specific rotation: [\u03b1]D20=+7.338\u00b0\u2005mL\u2009g\u22121\u2009dm\u22121 .Elemental analysis: Calc: C: 67.17\u2009%, H: 8.74\u2009%, N: 3.92\u2009%. Found: C: 67.3\u2009%, H: 8.7\u2009%, N: 4.0\u2009%.Compound \u20106: \u20101 was dissolved in 1\u2005mL acetone. ZnBr2 was dissolved in 1\u2005mL acetone and added to the silanol solution. After a few days, the product \u20106 crystallized from acetone in the form of colorless blocks in space group P212121 . Yield: 0.06\u2005g, 0.07\u2005mmol Elemental analysis: Calc.: C: 52.03\u2009%, H: 6.14\u2009%, N: 3.28\u2009%. Found: C: 52.3\u2009%, H: 6.2\u2009%, N: 3.4\u2009%.The authors declare no conflict of interest.1As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "The triazole ring in the title mol\u00adecule is not quite coplanar with the six-membered ring to which it is fused, the dihedral angle between the two least-squares planes being 2.52\u2005(6)\u00b0. In the crystal, a layer structure is formed by N\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds plus slipped \u03c0-stacking inter\u00adactions, with the fused cyclo\u00adhexene rings projecting to either side. 9H10N4O, is not quite coplanar with the six-membered ring to which it is fused, the dihedral angle between the two least-squares planes being 2.52\u2005(6)\u00b0. In the crystal, a layered structure is formed by N\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds plus slipped \u03c0-stacking inter\u00adactions, with the fused cyclo\u00adhexene rings projecting to either side.The triazole ring in the title mol\u00adecule, C Karan et al., 2021e.g. Zhao et al., 2021e.g. El-Shershaby et al., 2021e.g. Zhang et al., 2020e.g. Ibrahim et al., 2021Compounds containing nitro\u00adgen heterocycles make up a significant portion (approximately 60%) of small drug mol\u00adecules that have been approved by the FDA \u2005\u00c5, \u03b8 = 129.71\u2005(14)\u00b0 and \u03c6 = 326.36\u2005(18)\u00b0. This conformation is quite similar to a half-chair form. The C8/N2/C9/N3/N4 ring is closer to planarity than is the C1/C2/C7/N1/C8/N4 ring and the dihedral angle between their mean planes is 2.52\u2005(6)\u00b0. In the crystal, N1\u2014H1\u22efN3 hydrogen bonds and ethyl 2-oxo-cyclo\u00adhexa\u00adnecarboxyl\u00adate were combined and heated under reflux in 10\u2005ml of acetic acid for 1\u2005h. The solid product obtained was recrystallized from ethanol solution to afford colorless crystals.1Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314623004091/tk4091sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314623004091/tk4091Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623004091/tk4091Isup3.cmlSupporting information file. DOI: 2259950CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The distorted coordination octa\u00adhedra around Li+ involve four short bonds with two pyrazolate N atoms and two aqua ligands and two longer contacts with nitro-O atoms. When combined with \u03bc4-{TNBPz}2\u2212, this generates a mono-periodic polymeric structure incorporating discrete centrosymmeric [(H2O)2Li\u2013(di\u00adnitro\u00adpyrazolato)2\u2013Li(H2O)2] units. The three-dimensional stack of mutually orthogonal coordination chains is reminiscent of a Lincoln log pattern.In the title salt, the 3,3\u2032,5,5\u2032-tetra\u00adnitro-4,4\u2032-bi\u00adpyrazole-1,1\u2032-diide dianion [{TNBPz} 2(C6N8O8)(H2O)4]n, the 3,3\u2032,5,5\u2032-tetra\u00adnitro-4,4\u2032-bi\u00adpyrazole-1,1\u2032-diide dianion [{TNBPz}2\u2212] is situated across the twofold axis. The distorted coordination octa\u00adhedra around Li+ involve four short bonds with two pyrazolate N atoms and two aqua ligands [Li\u2014N(O) = 1.999\u2005(3)\u20132.090\u2005(2)\u2005\u00c5] and two longer contacts with nitro-O atoms . When combined with \u03bc4-{TNBPz}2\u2212, this generates a mono-periodic polymeric structure incorporating discrete centrosymmeric [(H2O)2Li\u2013(di\u00adnitro\u00adpyrazolato)2\u2013Li(H2O)2] units. The three-dimensional stack of mutually orthogonal coordination chains is reminiscent of a Lincoln log pattern. It is influenced by conventional hydrogen bonding [O\u22efO = 2.8555\u2005(17)\u20133.0010\u2005(15)\u2005\u00c5] and multiple lone pair\u2013\u03c0 hole inter\u00adactions of the nitro groups [N\u22efO = 3.0349\u2005(15) and 3.0887\u2005(15)\u2005\u00c5]. The Hirshfeld surface and two-dimensional fingerprint plots also support the significance of non-covalent bonding. Coordinative saturation and a favorable geometry at the Li+ ions, dense packing of the polymeric subconnectivities and particularly extensive inter\u00adanion inter\u00adactions may be involved in the stabilization of the structure. The title salt is a rare example of an energetic Li nitro\u00adazolate, which nicely crystallizes from aqueous solution and is neither hygroscopic nor efflorescent. The TG/DTA data reveal total dehydration in the range of 330\u2013430\u2005K and stability of the anhydrous material up to 633\u2013653\u2005K.In the structure of the title salt, [Li The nitro\u00adpyrazolates crystallize with difficulty and 2.090\u2005(2)\u2005\u00c5], two aqua-O atoms [1.999\u2005(3) and 2.027\u2005(3)\u2005\u00c5] and two elongated bonds with nitro-O atoms [2.550\u2005(2) and 2.636\u2005(2)\u2005\u00c5] Li]n\u2013 \u2005\u00c5 [symmetry code: (ii) \u2212x\u00a0+\u00a0y\u00a0+\u00a0z], are likely attractive, as a kind of lone pair\u2013\u03c0 hole bonding \u2005\u00c5 vs 1.4644\u2005(15) and 1.462\u2005(2)\u2005\u00c5 for H2(TNBPz) and Rb{H(TNBPz)}, respectively], which indicates a lack of essential conjugation between two pyrazolate rings. Shortening of the C-NO2 bonds upon deprotonation is also minor [mean 1.4308\u2005(16)\u2005\u00c5 vs 1.439\u2005(2)\u2005\u00c5 for H2(TNBPz)], while a certain increase in conjugation is reflected rather by a perceptible flattening of the di\u00adnitro\u00adpyrazole fragments. For the latter, the NO2 groups are nearly coplanar with the ring, the two dihedral angles are 1.35\u2005(8) and 11.66\u2005(8)\u00b0, for N4O3O4 and N3O1O2 groups, respectively. In the case of H2(TNBPz), the twist comes to 22.8\u2005(2)\u00b0. The most appreciable consequence of the dianionic structure is similarity of bond angles at the ring-N atoms: N2\u2014N1\u2014C1 = 107.11\u2005(9)\u00b0 and N1\u2014N2\u2014C3 = 106.93\u2005(9)\u00b0. For the neutral di\u00adnitro\u00adpyrazole rings, the parameters for N- [103.9\u2005(2)\u00b0] and NH-sites [112.0\u2005(2)\u00b0] are clearly different 2 cycles (pz is pyrazole).The title compound adopts a mono-periodic polymeric structure with the {TNBPz}rn Fig.\u00a02, which iA\u22efO6vii are important for the connection of the chains into layers, which are parallel to the ab plane, whereas the second aqua/aqua bond O5\u2014H\u22efO6v is readily available by nitration of 4,4\u2032-bi\u00adpyrazole in mixed acids (yield 92%) and subsequent crystallization from water of solid H2(TNBPz)\u00b7H2O was added with stirring. The mixture was stirred for 30\u2005min and the resulting clear deep-yellow solution was cooled to r.t. Slow evaporation to a minimum volume over 8\u201310\u2005d led to crystallization of the product as well-developed large lemon-yellow prisms. The crystals were removed and dried on a filter paper in air. The yield was 1.25\u2005g (90%). The material shows neither signs of hygroscopy nor efflorescence when exposed to ambient air for months.For the preparation of the title compound, 0.294\u2005g (7.0\u2005mmol) of LiOH\u00b7H6H8Li2N8O12: C 18.10, H 2.03, N 28.15; found: C 18.45, H 1.99, N 28.42. IR : 564 m, 636 m, 698 m, 716 m, 773 w, 853 s, 1006 m, 1021 s, 1189 w, 1284 w, 1308 s, 1321 s, 1353 s, 1381 s, 1410 s, 1484 s, 1540 s, 1641 m, 1658 m, 3460 br, 3580 br.Analysis (%) calculated for C\u22121 (O\u2014H stretching), 1641, 1658\u2005cm\u22121 (bend) and 564\u2005cm\u22121 (libration). The peaks for symmetric and asymmetric NO2 stretching are very similar to the spectra of comparable 3,5-di\u00adnitro\u00adpyrazole , and its shift, with respect to the band for H2(TNBPz)\u00b7H2O . The anhydrous material is stable up to 633\u2005K, with the very minor exothermic event at 597\u2005K possibly indicating a phase transition. Exothermic decomposition proceeds above 653\u2005K, with instantaneous loss of any remaining weight and a sharp exothermic effect at ca 700\u2005K suggesting an explosion. For comparison, typical onset temperatures for decomposition of energetic Li nitro\u00adpyrazolates are 400\u2013500\u2005K, and only 3,5-di\u00adnitro\u00adpyrazolate is stable up to 600\u2005K . There are two partially separated stages for nearly identical weight losses in the temperature range of 330\u2013430\u2005K Fig.\u00a07, which c6.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023005339/pk2688sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989023005339/pk2688Isup2.hklStructure factors: contains datablock(s) I. DOI: 2269963CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The caffeinium cations form a double layer via hydrogen bonds and \u03c0-stacking inter\u00adactions. The Hirshfeld surface analysis showed that the largest contribution to the crystal packing is made by H\u22efH (31.2%), H\u22efCl/Cl\u22efH (22.6%), O\u22efH/H\u22efO (21.9%) contacts for the cation and H\u22efCl/Cl\u22efH (79.3%) contacts for the anion.The mol\u00adecular and crystal structure of the title compound, (C Studies on the inter\u00adaction of hexa\u00adchloro\u00adplatinates with various biological organic compounds have been performed before, for example by Novikov et al. hexa\u00adchloro\u00adplatinum RECJAO crystallizes in the triclinic space group Pb) contains two caffeinium cations and one centrosymmetric hexa\u00adchloro\u00adplatinate anion with a platinum atom in a special position 1a. In the imidazole ring of the caffeine mol\u00adecule, the nitro\u00adgen N1 atom is protonated. The cation, including the methyl groups, has a flat geometry (maximum deviation for non-hydrogen atoms 0.030\u2005\u00c5). The [PtCl6]2\u2212 anion has a slightly distorted octa\u00adhedral geometry with similar Pt\u2014Cl bond distances \u2005\u00c5 .Hydrogen bonds and \u03c0-stacking play a significant role in the formation of inter\u00admolecular inter\u00adactions in the crystal structure of \u2005\u00c5 Fig.\u00a01b.et al., 2016et al., 2018et al., 2011et al., 2022Cz) and chlorine atoms Cl1 and Cl2, with Cz \u22ef Cl distances of 3.8643\u2005(11) and 3.7170\u2005(11)\u2005\u00c5, respectively, and \u03b1 angles between the ring plane and the Cz\u22efCl vector of 61.82\u2005(7) and 62.28\u2005(7)\u00b0, respectively. It is uncertain whether such an inter\u00adaction exists with Cl3 .Similarly, \u03c0\u2013halogen inter\u00adactions plane \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z] inter\u00adaction being the strongest.The crystal packing in ne Fig.\u00a02. The cafne Fig.\u00a02, the N1\u20144.Crystal Explorer 21 was used to calculate the Hirshfeld surfaces (HS) and two-dimensional fingerprint plots to 1.063 (blue) for cation and \u22120.402 to 0.934 a.u. for anion, as illus\u00adtrated in Fig.\u00a03b).s Figs. 3 and 4 \u25b8.c,d,i. The structures of II and III show distributions of contacts .Analysis of inter\u00admolecular contacts shows that for the caffeinum cation, the largest contributions are made by H\u22efH, Cl\u22efH/H\u22efCl and O\u22efH/H\u22efO contacts Fig.\u00a05, and fors Figs. 5 and 6 \u25b8 5.et al., 20166 type. The closest analogues of I found were II and III (see above), the former containing N-protonated 3-carb\u00adoxy\u00adpyridine (nicotinic acid) as the cation and [PtCl6]2\u2212 as the anion, the latter containing a caffeinium cation with a methyl\u00adated (rather than protonated) N1 atom and a PF6\u2212 anion.A search of the Cambridge Structural Database refined], C4H3 as rotationally disordered between two orientations with occupancies of 0.62\u2005(4) and 0.38\u2005(4) [Uiso(H) = 1.2Ueq(C)], with C\u2014H 0.96\u2005\u00c5 in each case. The H atoms at N1 and C3 were refined isotropically.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023005157/zv2027sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023005157/zv2027Isup2.hklStructure factors: contains datablock(s) I. DOI: 2268577CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II cinnamaldehyde 4-thio\u00adsemicarbazone homoleptic complex is reported. As a result of H\u22efS intra\u00admolecular inter\u00adactions, graph-set motif S(5), the coordination sphere resembles a hydrogen-bonded macrocyclic environment-type. In the crystal, the mol\u00adecules are linked by H\u22efS inter\u00adactions, with graph-set motifs The synthesis, crystal structure and Hirshfeld surface analysis of a new Pd E)-N-phenyl-2-[(2E)-3-phenyl-2-propen-1-yl\u00adidene]hydra\u00adzine\u00adcarbo\u00adthio\u00adamide deprotonated with NaOH in ethanol with an ethano\u00adlic suspension of PdII chloride in a 2:1 molar ratio yielded the title compound, [Pd(C16H14N3S)2]. The anionic ligands act as metal chelators, \u03ba2N1S-donors, forming five-membered rings with a trans-configuration. The PdII ion is fourfold coordinated in a slightly distorted square-planar geometry. For each ligand, one H\u22efS and one H\u22efN intra\u00admolecular inter\u00adactions are observed, with S(5) and S(6) graph-set motifs. Concerning the H\u22efS inter\u00adactions, the coordination sphere resembles a hydrogen-bonded macrocyclic environment-type. In the crystal, the complexes are linked via pairs of H\u22efS inter\u00adactions, with graph-set motif R22(8), and building a mono-periodic hydrogen-bonded ribbon along [001]. The Hirshfeld surface analysis indicates that the major contributions for the crystal cohesion are: H\u22efH (45.3%), H\u22efC/C\u22efH (28.0%), H\u22efS/S\u22efH (8.0%) and H\u22efN/N\u22efH (7.4%).The reaction of (2 It was pointed out that the main product of the characterization reaction was a thio\u00adsemicarbazone derivative, R3R4C=N\u2014N(H)C(=S)NR1R2 \u2014C(=S) fragment can be easily deprotonated and the negative charge is then delocalized over the N\u2014N\u2014C\u2014S entity, which enables chemical bonding with many different metal centers, with different Lewis acidity, and a diversity of coordination modes, II heteroleptic complexes with a cinnamaldehyde-thio\u00adsemicarbazone deriv\u00adative turned out to be very active on in vitro Human Topo\u00adisomerase II\u03b1 inhibition, a biological target of prime importance for cancer research , cervix (HeLa), hepatocellular (HepG2), breast (MCF-7) and prostate (PC-3) and lung (A549) 2 homo- and heteroleptic complexes results in the formation of ZnS nanocrystallites 2 and CdI2(LH)2 were used as starting materials to obtain CdS nanoparticles 2 homo- and heteroleptic complexes and 177.57\u2005(2)\u00b0, respectively. The metal ion is fourfold coordinated in a slightly distorted square-planar geometry. The maximum deviation from the mean plane through the Pd1/N1/N4/S1/S2 fragment is 0.0227\u2005(5)\u2005\u00c5 for Pd1 and the r.m.s. for the selected atoms is 0.0151\u2005\u00c5. Concerning the geometry of the N\u2014N\u2014C\u2014S entities, the N1\u2014N2\u2014C10\u2014S1 torsion angle is 0.6\u2005(3)\u00b0, while N4\u2014N5\u2014C26\u2014S2 is \u22120.4\u2005(3)\u00b0. Both of the ligands are non-planar, with the angle between the mean planes through the C4\u2013C9 and the C11\u2013C16 aromatic rings being 15.7\u2005(1)\u00b0, while that between the C20\u2013C25 and the C27\u2013C32 rings is 45.5\u2005(8)\u00b0.The asymmetric unit comprises one mol\u00adecule of the title compound, with all atoms being located in general positions Fig.\u00a01. The comS(5), and C16\u2014H13\u22efN2 and C32\u2014H26\u22efN5, with graph-set motif S(6). Considering the S(5) rings, a hydrogen-bonded macrocyclic coordination environment-type can be suggested for the PdII metal center, while the S(6) rings contribute to the stabilization of the mol\u00adecular structure.Four intra\u00admolecular hydrogen-bonding inter\u00adactions are observed Fig.\u00a02: C1\u2014H1\u22efSi.e., non-coordinating thio\u00adsemicarbazones, the N\u2014N and C\u2014S bonds have lengths of double-bond character, while the N\u2014C bond shows lengths of single-bond type, which can be written as a N=N(H)\u2014C=S fragment. When the acidic H atom of the hydrazinic fragment is removed, the negative charge is delocalized over the N\u2014N\u2014C\u2014S chain and the bond lengths change to inter\u00admediate values. Thus, the N\u2014N and the C\u2014S bond lengths assume single-bond character, being longer, and the N\u2014C bond lengths assume double-bond character, being shorter. Information about the bond lengths of the N\u2014N\u2014C\u2014S entities for the cinnamaldehyde-4-phenyl\u00adthio\u00adsemicarbazone mol\u00adecule, C16H15N3S, and the Ni(C16H14N3S)2 2 complexes, this work, are summarized in Table\u00a02et al., 2014Finally, the anionic form of the ligands was assigned because of the absence of hydrazinic H atoms and the change in the bond lengths of the N\u2014N\u2014C\u2014S entities. For the neutral or free, 3.via pairs of N\u2014H\u22efS inter\u00adactions with graph-set motif In the crystal, the mol\u00adecules are connected 1] Fig.\u00a03.Crystal Explorer x, \u2212y\u00a0+\u00a0z\u00a0+\u00a0x, \u2212y\u00a0+\u00a0z\u00a0\u2212\u00a0a) H\u22efH (45.3%), (b) H\u22efC/C\u22efH (28.0%), (c) H\u22efS/S\u22efH (8.0%) and (d) H\u22efN/N\u22efH (7.4%). The contributions to the crystal packing are shown as two-dimensional Hirshfeld surface fingerprint plots with cyan dots and the de (y-axis) values are the closest inter\u00adnal and external distances from given points on the Hirshfeld surface (in \u00c5).The Hirshfeld surface analysis and as a ligand, viz. in the homoleptic [Ni(C16H14N3S)2] complex \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01]. The negative charge of the ligand was assigned by the absence of a hydrazinic H atom and the bond distances in the N\u2014N\u2014C\u2014S chain , the metal center is fourfold coordinated in a square-planar geometry and the N\u2014N\u2014C\u2014S entity torsion angle is 1.5\u2005(6)\u00b0.To the best of our knowledge and using database tools such as ry Fig.\u00a06 Fig.\u00a06. The H\u22efC] Fig.\u00a06 and theyII coordination compound was also performed with CrystalExplorer H\u22efH (47.4%), (b) H\u22efC/C\u22efH (27.6%), (c) H\u22efN/N\u22efH (7.0%) and (d) H\u22efS/S\u22efH (6.5%). The di (x-axis) and the de (y-axis) values are the closest inter\u00adnal and external distances from given points on the Hirshfeld surface contacts (in \u00c5). While for the PdII title compound and the NiII reference compound the most important inter\u00admolecular contacts are H\u22efH and the H\u22efC/C\u22efH, the order of importance changes for the H\u22efS/S\u22efH and H\u22efN/N\u22efH contacts. For the crystal packing of the PdII complex, the H\u22efS/S\u22efH contacts are more important then H\u22efN/N\u22efH contacts, while for the NiII complex this order is the opposite.The Hirshfeld surface analysis = 1.2 Ueq, and with C\u2014H = 0.93\u2005\u00c5 and N\u2014H = 0.86\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023008654/zn2032sup1.cifCrystal structure: contains datablock(s) I, publication_text. DOI: 10.1107/S2056989023008654/zn2032Isup2.hklStructure factors: contains datablock(s) I. DOI: 2163054CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, pairs of mol\u00adecules are linked by N\u2014H\u22efN hydrogen bonds, forming 6H8N2OS, all atoms except for the methyl H atoms are coplanar, with a maximum deviation of 0.026\u2005(4)\u2005\u00c5. In the crystal, pairs of mol\u00adecules are linked by N\u2014H\u22efN hydrogen bonds, forming R22(8) ring motifs. Dimers are connected by N\u2014H\u22efO hydrogen bonds, forming layers parallel to the (102) plane. Consolidating the mol\u00adecular packing, these layers are connected by C\u2014H\u22ef\u03c0 inter\u00adactions between the center of the 1,3-thia\u00adzole ring and the H atom of the methyl group attached to it, as well as C=O\u22ef\u03c0 inter\u00adactions between the center of the 1,3-thia\u00adzole ring and the O atom of the carboxyl group. According to a Hirshfeld surface study, H\u22efH (37.6%), O\u22efH/H\u22efO (16.8%), S\u22efH/H\u22efS (15.4%), N\u22efH/H\u22efN (13.0%) and C\u22efH/H\u22efC (7.6%) inter\u00adactions are the most significant contributors to the crystal packing.In the title compound, C Other major contributors are O\u22efH/H\u22efO , S\u22efH/H\u22efS , N\u22efH/H\u22efN and C\u22efH/H\u22efC inter\u00adactions. Other, smaller contributions are made by S\u22efC/C\u22efS (2.7%), C\u22efO/O\u22efC (2.6%), C\u22efC (1.8%), N\u22efC/C\u22efN (1.5%), S\u22efO/O\u22efS (0.8%), S\u22efN/N\u22efS (0.1%) and O\u22efN/N\u22efO (0.1%) inter\u00adactions.u. Fig.\u00a07. The intu. Fig.\u00a07b. Other4.et al., 20161,3-thia\u00adzole yielded five compounds related to the title compound, viz. CSD refcodes IXAMAV via weak C\u2014H\u22efO hydrogen bonds into a two-dimensional supra\u00admolecular framework. The crystal structure of DUTZEY involves inter\u00admolecular N\u2014H\u22efN hydrogen bonds. In the crystal of LAMQOJ, weak C\u2014H\u22efN hydrogen bonds build up a wavy layer of mol\u00adecules in the (011) plane. The layers are stacked in the [100] direction by weak \u03c0\u2013\u03c0 stacking inter\u00adactions between the 1,3-thia\u00adzole rings.In the crystal of IXAMAV, the supra\u00admolecular network is based upon N\u2014H\u22efN hydrogen-bonded centrosymmetric dimers linked by N\u2014H\u22efO contacts. ABEGAQ forms a supra\u00admolecular network based on N\u2014H\u22efN hydrogen-bonded centrosymmetric dimers that are linked in turn by N\u2014H\u22efO contacts. In the crystal of FEFKUY, an inter\u00adplay of O\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds connects the mol\u00adecules to form 5.et al., 2012The title compound was synthesized using a reported procedure = 1.2Ueq(N) for the NH2 group and 1.5Ueq(C) for CH3 groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023007181/vm2288sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023007181/vm2288Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023007181/vm2288Isup3.cmlSupporting information file. DOI: 2288949CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The packing involves three borderline C\u2014H\u22efN contacts and a pairing of the triazole rings across an inversion centre.In the crystal structure of the title compound, the triazole ring exhibits inter\u00adplanar angles of 17H14N4O2, the triazole ring exhibits inter\u00adplanar angles of 63.86\u2005(2) and 76.96\u2005(2)\u00b0 with the phenyl and benzo\u00adthia\u00adzole planes, respectively. The C\u2014C\u2014C angle at the methyl\u00adene group is rather wide at 114.28\u2005(4)\u00b0. The packing involves three borderline C\u2014H\u22efN contacts, two of which combine to form layers parallel to ac, and a pairing of the triazole rings across an inversion centre [inter\u00adplanar distance of 3.1852\u2005(2)\u2005\u00c5].In the structure of the title compound, C A selection, mostly involving the heteroatoms, is presented in Table\u00a01imine contact in N-[3-(benzo[d]thia\u00adzol-2-yl)-6-bromo-2H-chromen-2-yl\u00adidene]-4-methyl\u00adbenzenamine , 3.1852\u2005(2) and 0.94\u2005\u00c5, respectively. This feature is reinforced by the other C\u2014H\u22efN inter\u00adaction, which involves the same operator.The mol\u00adecular packing displays few significant features. There are three borderline C\u2014H\u22efN inter\u00adactions Table\u00a02, two of ne Fig.\u00a03. The tri4.et al., 2002et al., 2016The searches employed the routine ConQuest -1,3-benzo\u00adthia\u00adzole to 445 values in the range 106\u2013122\u00b0, with a mean value of 114\u2005(5)\u00b0. However, restricting one ring to be a C2-substituted thia\u00adzole gave only three hits, with four values of 109.6\u2013112.9\u00b0 for the angle at the methyl\u00adene groups. These all involved two planar ring systems of the benzo[d]thia\u00adzole type, but with different heteroatoms in some cases and phenyl iso\u00adthio\u00adcyanate 2 (0.01\u2005mol) was stirred for 30\u2005min in ethanol (25\u2005mL) in the presence of sodium ethoxide (0.01\u2005mol). After cooling, methyl iodide (0.015\u2005mol) was added. The reaction mixture was stirred for 30\u2005min at room temperature, then refluxed for 1\u2005h. The resulting precipitate was filtered off, washed with water, dried, and recrystallized from ethanol. The title compound was isolated as a white solid; yield 75%; m.p. 429\u2005K; IR : \u03bd 3053 (Ar\u2014CH), 2928 , 1594 (C=N); 1H NMR : \u03b4 2.60 , 4.57 , 7.36\u20137.50 , 7.89 , 8.01 ; Analysis calculated for C17H14N4S2 (338.45): C 60.33, H 4.17, N 16.55, S 18.95. Found C 60.66; H 4.15; N 16.40; S 18.90%.A mixture of 2-benzo\u00adthia\u00adzolyl acetohydrazide 6.aromatic = 0.95\u2005\u00c5, C\u2014Hmethyl\u00adene = 0.99\u2005\u00c5). The U(H) values were fixed at 1.5 \u00d7 Ueq of the parent carbon atoms for the methyl group and 1.2 \u00d7 Ueq for other hydrogens.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023007041/yz2038sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989023007041/yz2038Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023007041/yz2038Isup3.cmlSupporting information file. DOI: 2287438CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure of the title bis-urea derivative, the host mol\u00adecules are linked by N\u2014H\u22efO=C hydrogen bonds and C\u2014H\u22efO contacts with 26H26N4O6\u00b72C4H8O, representing a bis-urea with terminal phenyl\u00adalanine units, crystallized with two tetra\u00adhydro\u00adfuran (THF) mol\u00adecules. The main mol\u00adecule is located on a crystallographic twofold axis, while the solvent mol\u00adecule is disordered over two positions, with occupancies of 0.571\u2005(15) and 0.429\u2005(15). The host mol\u00adecules are linked by N\u2014H\u22efO=C hydrogen bonds and C\u2014H\u22efO contacts with R21(6) and R21(7) ring motifs. The THF mol\u00adecules enclosed in the crystal are connected to the bis-urea compound via O\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions.The title compound, C The angle between the planes of adjacent urea moieties is 83.7\u2005(1)\u00b0, thus they are nearly perpendicular to one other. Such a motif is also well known in the literature and 2.32\u2005(3)\u2005\u00c5; Table\u00a013 (corresponding to about 29% of the unit-cell volume). The cavities are bounded by the nonpolar phenyl\u00adene and arene units of the title compound. In addition, the carb\u00adoxy groups point into the inter\u00adior of these cavities and form O\u2014H\u22efO hydrogen bonds with the THF O atom [H\u22efO = 1.73\u2005(3)\u2005\u00c5]. Further stabilization of the mol\u00adecular network, each involving the THF mol\u00adecules, is realized by C\u2014H\u22efO contacts with the carb\u00adoxy group of an adjacent mol\u00adecule (H\u22efO = 2.78\u2005\u00c5) and weak C\u2014H\u22ef\u03c0 contacts (H\u22efCg = 2.61\u20133.00\u2005\u00c5) with the central benzene core or peripheral arene substituents.The crystal structure exhibits cavities which are occupied by THF mol\u00adecules requiring about 961\u2005\u00c5et al., 2012The synthetic and spectroscopic details for the title compound have been reported previously (Stapf Uiso(H) values were fixed at 1.2 times the equivalent Ueq value of the parent C atoms. The THF solvent mol\u00adecule is disordered over at least two positions [refined occupancies 0.571\u2005(15) and 0.429\u2005(15)]. Therefore, the solvent mol\u00adecule was refined using ISOR for C16A, C16B, C17A and C17B (approximate isotropic behaviour) and SADI (same distances over pairs of bonded atoms) restraints I. DOI: 10.1107/S2414314623007435/bh4077Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623007435/bh4077Isup3.cmlSupporting information file. DOI: 2290568CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The first high-quality crystal structure of unmodified 2\u2032-de\u00adoxy\u00adguanosine is reported. The isolated crystals are the dimethyl sulfoxide disolvate. 10H13N5O4\u00b72C2H6OS, which is of inter\u00adest with respect to its biological activity, at 183\u2005K has ortho\u00adrhom\u00adbic (P212121) crystal symmetry. The structure displays a network of inter\u00admolecular N\u2014H\u22efN, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds. 2\u2032-De\u00adoxy\u00adguanosine mol\u00adecules are linked to each other and to the two dimethyl sulfoxide solvent mol\u00adecules by hydrogen bonding.The title com\u00adpound, C Understanding the properties of DNA is crucial for our knowledge of its reactivity in cellular processes of replication and transcription to yield transfer RNA crystallized in the ortho\u00adrhom\u00adbic Sohnke space group P212121, with four formula units per unit cell and one per asymmetric unit .Nucleobase -one. The cocrystal structures of (actinomycin D)\u00b72(2\u2032-de\u00adoxy\u00adguanosine)\u00b712H2O (ACTDGU) and (7-bromo\u00adac\u00adtino\u00admycin)\u00b72(2\u2032-de\u00adoxy\u00adguanosine)\u00b711H2O (BRAXGU) at room temperature were also reported , is the only purine nucleoside solvate in the CSD were obtained upon slow evaporation of 2\u2032-de\u00adoxy\u00adguanosine from DMSO.Single crystals of values were set at 1.2 times or 1.5 times (for methyl and OH groups) the Ueq value of the parent atom. The Flack parameter x was \u22120.00\u2005(6) by classical fit to all intensities and 0.022\u2005(14) by Parsons\u2019 method I, global. DOI: 10.1107/S2056989023007405/zv2028Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989023007405/zv2028sup3.pdfFigures S1 and S2. DOI: 2290127CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular conformation is stabilized by two intra\u00admolecular N\u2014H\u22efN inter\u00adactions. In the crystal, mol\u00adecules are also linked via C\u2014H\u22ef\u03c0 inter\u00adactions forming a three-dimensional network.In the title compound, [24]di\u00adthia\u00adamethyrin(1.0.0.1.0.0); C 50H44Cl4N4S2 were obtained from the reaction of 2,5-bis\u00ad(4-propyl-1H-pyrrol-2-yl)thio\u00adphene and 3,5-di\u00adchloro\u00adbenzaldehyde in the presence of tri\u00adfluoro\u00adacetic acid for 3\u2005h and subsequent addition of p-chloranil. The macrocycle in the title compound can be described as a highly planar structure wthe the average deviation of the 32 macrocyclic atoms from the least-squares plane being 0.0416\u2005\u00c5. Its mol\u00adecular conformation is stabilized by two intra\u00admolecular N\u2014H\u22efN bonds and a three-dimensional network is formed by C\u2014H\u22ef\u03c0 inter\u00adactions.Purple crystals of the title compound, C Neighboring mol\u00adecules form dimers via inter\u00admolecular C\u2014H\u22efCl and C\u2014H\u22ef\u03c0 inter\u00adactions owing to the H6\u22efCl1 (2.93\u2005\u00c5), H17A\u22efCl2 (3.34\u2005\u00c5), and C3\u2014C11 (3.36\u2005\u00c5) distances, as shown in Fig.\u00a02The mol\u00adecular structures and electronic properties of hexa\u00adpyrrolic expanded porphyrins with different numbers of \u03c0-electrons and et al. (20152Cl2 (ca 300\u2005mL) under an Ar atmosphere, to which 3,5-di\u00adchloro\u00adbenzaldehyde (68.4\u2005\u00b5L) and tri\u00adfluoro\u00adacetic acid (160\u2005\u00b5L) were added. The reaction mixture was stirred for 3\u2005h, p-chloranil (544\u2005mg) was added to it, and the mixture was stirred overnight at ambient temperature. Then, the mixture was neutralized with an aqueous NaHCO3 solution, and the crude products were passed through an alumina column. Finally, the products were purified by chromatography on a silica gel column using chloro\u00adform as elute. The third blue (2%) fraction afforded the title compound. The compound was recrystallized from a mixture of hexane and chloro\u00adform. Purple plates of suitable quality for diffraction were obtained by slow-diffusing hexane into chloro\u00adform. 1H NMR : \u03b4 (ppm) 24.0 , 7.20\u2013 6.87 , 5.05 , 4.61 , 0.76 , 0.52 , 0.32 ; MALDI\u2013TOF MS found = 904.254, monoisotopic mass = 904.176 calculated for C50H44Cl4N4S2. UV\u2013vis (CH2Cl2): \u03bb = 387, 500\u2005nm.The title compound was prepared by a modified previously reported method by Ishimaru al. 2015. 2,5-BisCrystal data, data collection, and structure refinement details are summarized in Table\u00a0210.1107/S2414314623007666/xu4052sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314623007666/xu4052Isup2.hklStructure factors: contains datablock(s) I. DOI: 2292415CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, chains of mol\u00adecules extending along the 28H34ClN3O9S, is disordered over two closely spaced sets of sites; the site occupancy of the major component = 0.542\u2005(3). The conformation of each component is approximately U-shaped with the chloro\u00adbenzene ring forming the base and the indolinyl and sulfamoyl groups the sides; an intra\u00admolecular C\u2014H\u22efCl hydrogen bond possibly contributes to the stabilization of the conformation. In the crystal, a corrugated layer structure parallel to the ab plane is formed by C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds together with C\u2014H\u22ef\u03c0(ring) inter\u00adactions.The majority of the title mol\u00adecule, C Missioui et al., 2022Indapamide, is a di\u00adhydro-indole-based thia\u00adzide-like diuretic used to manage heart failure and treat hypertension. Various activities are associated with indole derivatives, including anti\u00adviral = 0.247\u2005(8)\u2005\u00c5 and \u03c6(2) = 327\u2005(2)\u00b0 for the major component and Q(2) = 0.399\u2005(9)\u2005\u00c5 and \u03c6(2) = 329.6\u2005(16)\u00b0 for the other. The dihedral angle between the mean planes of the C1\u2013C6 and C15\u2013C20 rings is 86.2\u2005(4)\u00b0 while that between the two disordered components of the C1\u2013C6 ring is 3.8\u2005(5)\u00b0 and that between the two disordered components of the C15\u2014C20 ring is 5.68\u2005(6)\u00b0.The mol\u00adecule adopts an approximate U-shaped conformation Fig.\u00a01 with thed Table\u00a01 may helpd Table\u00a01 of the tA\u22efCl1 hydrogen bonds and potassium bicarbonate were dissolved in di\u00admethyl\u00adformamide (10\u2005mL), to which was added ethyl 2-bromo\u00adacetate . Under reflux, the reaction was stirred for 3\u2005h at 80\u00b0C. When the starting reagents had reacted completely, distilled water (100\u2005ml) was added. The product precipitated in solid form, was filtered, dried and recrystallized from ethanol solution to afford colorless blocks.N-(2-methyl\u00adindolin-1-yl)glycinate portion of the mol\u00adecule is disordered over two partially resolved sets of sites in a 0.542\u2005(3):0.458\u2005(3) ratio. In addition, the C23\u2014C24 ethyl group is disordered over two sets of sites in a 0.526\u2005(12):0.474\u2005(12) ratio. The two components of each disorder were refined with restraints that their geometries be comparable.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314623006995/tk4094sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314623006995/tk4094Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623006995/tk4094Isup3.cmlSupporting information file. DOI: 2280340CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title hydrated salt features a dense array of hydrogen bonds, forming a three-dimensional network. 5H5Cl2N2+\u00b7C6H4NO3\u2212\u00b7H2O, the pyridine N atom of the cation is protonated and an intra\u00admolecular O\u2014H\u22efO hydrogen bond is observed in the anion, which generates an S(6) ring. The crystal packing features N\u2014H\u22efN, O\u2014H\u22efO, N\u2014H\u22efO, C\u2014H\u22efCl and C\u2014H\u22efO hydrogen bonds, which generate a three-dimensional network.In the title hydrated salt, C In the extended structure, the cations, anions and water mol\u00adecules are connected by N\u2014H\u22efN, O\u2014H\u22efO, C\u2014H\u22efCl, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds was mixed with a hot aqueous solution of 4-amino 3,5-di\u00adchloro pyridine (34\u2005mg). The mixture was cooled slowly and kept at room temperature. After a few days, colourless block shaped crystals were obtained.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314623008210/hb4452sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314623008210/hb4452Isup2.hklStructure factors: contains datablock(s) I. DOI: 2294939CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "To provide the correct values of UCCR, the UCCR was calculated again by converting cortisol and creatinine in the same measurement unit (nmol/L).Section 3 Analytical Procedures, first paragraph, last sentence, should be as follows. \u201cThe UCCR was calculated by dividing the urine cortisol concentration (nmol/L) by the urine creatinine concentration (mmol/L).\u201dReference 29 is removed from the References list.\u22126 (1.04\u20103.81\u2009\u00d7\u200910\u22126), 32.09\u2009\u00d7\u200910\u22126 (7.68\u2010245.5\u2009\u00d7\u200910\u22126), and 10.55\u2009\u00d7\u200910\u22126 (3.47\u201054.05\u2009\u00d7\u200910\u22126) in dogs with HA, dogs with DMHA and healthy dogs, respectively.\u201dSection 4 Results, UCCR subsection, first sentence should be as follows. \u201cThe median UCCR was 2.03\u2009\u00d7\u200910\u22126 (7.68\u2010245.46\u2009\u00d7\u200910\u22126).Section 4 Results, UCCR subsection, fifth sentence should be as follows. \u201cThe median UCCR in dogs with DMHA and BSC \u22642\u2009\u03bcg/dL (\u226455\u2009nmol/L) was 27.14\u2009\u00d7\u200910Section 4 Results, UCCR subsection, last sentence should be as follows. \u201cA cut\u2010off value of UCCR <4.4 revealed 100% sensitivity (95% CI: 69.1\u2010100) and 97.3% specificity (95% CI: 85.8\u201099.9) in diagnosing HA.\u201dFigure 1, to be replaced with this new Figure 1.FIGURE 1 Scatter scale plot comparing urinary corticoid: creatinine ratio (UCCR) of dogs with hypoadrenocorticism , dogs with disease mimicking hypoadrenocorticism and healthy dogs . The horizontal bars represent the median, the maximum, and the minimum value of each group."} +{"text": "The crystal structures of two benzo\u00adthio\u00adphene derivatives are described along with an analysis of the inter\u00admolecular contacts in the crystals performed using Hirshfeld surface analysis and two-dimensional fingerprint plots. 26H19NO2S2, (I), and C25H19NO2S2, (II), the benzo\u00adthio\u00adphene rings are essentially planar with maximum deviations of 0.026\u2005(1) and \u22120.016\u2005(1)\u2005\u00c5 for the carbon and sulfur atoms in compounds (I) and (II), respectively. In (I), the thio\u00adphene ring system is almost orthogonal to the phenyl ring attached to the sulfonyl group, subtending a dihedral angle of 88.1\u2005(1)\u00b0, and the di\u00adhydro\u00adpyridine ring adopts a screw\u2013boat conformation. In both compounds, the mol\u00adecular structure is consolidated by weak C\u2014H\u22efO intra\u00admolecular inter\u00adactions formed by the sulfone oxygen atoms, which generate S(5) ring motifs. In the crystal of II, mol\u00adecules are linked via C\u2014H\u22efO hydrogen bonds, generating C(7) chains running along the [100] direction. No significant inter\u00admolecular inter\u00adactions are observed in I.In both of the title compounds, C Thio\u00adphene derivatives possess pharmacological and biological activities including anti\u00adbacterial attached to an N--N-prop-2-yn-1-yl benzene\u00adsulfonamide group (C9\u2013C15/N1/S2/O1/O2) , 19.0\u2005(1) and 33.9\u2005(1), respectively, in compound I and 0.7\u2005(2), 38.1\u2005(2) and 87.6\u2005(2)\u00b0, respectively, in compound II with the C1\u2013C6, C11\u2013C16 and C17\u2013C22 phenyl rings. The benzo\u00adthio\u00adphene ring system is almost orthogonal to the C17\u2013C22 phenyl ring attached to the sulfonyl group in I, subtending a dihedral angle of 88.1\u2005(1)\u00b0, while the di\u00adhydro\u00adpyridine ring (C10/C11/C16/C23/C24) adopts a screw\u2013boat conformation, as is evident from the Cremer\u2013Pople puckering analysis of the six-membered heterocyclic ring The mol\u00adecular structure of compound I Fig.\u00a01 comprise2) Fig.\u00a02. In bothI and 119.9\u2005(2)\u00b0 in II], with a simultaneous decrease in the N1\u2014S2\u2014C17 angle [108.5\u2005(1)\u00b0 in I and 107.6\u2005(1)\u00b0 in II] from the ideal tetra\u00adhedral value (109.5\u00b0) are attributed to the Thorpe\u2013Ingold effect \u2005\u00c5 in II] and N1\u2014C16 . No significant inter\u00admolecular inter\u00adactions or C\u2014H\u22ef\u03c0 inter\u00adactions with centroid distances of less than 4\u2005\u00c5 are observed in the structure.In the crystal of II, mol\u00adecules are linked via C25\u2014H25\u22efO1 hydrogen bonding, generating C(7) chains \u2005\u00c5 where Cg3 is the centroid of the S1/C1/C6\u2013C8 ring; symmetry code: (ii) \u2212x, 2\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z] and C\u2014H\u22ef\u03c0 inter\u00adactions . are also present. Packing view of the title compound are shown in Figs. 3In the crystal of 4.et al., 2019et al., 2007dnorm, curvedness and shape index and 2D fingerprint plots) were performed with CrystalExplorer17 through white to blue . The dnorm surface was mapped over a fixed colour scale of \u22120.085 (red) to 1.564 (blue) for compound I and \u22120.286 (red) to 1.374 (blue) for compound II. The red spots indicate inter\u00admolecular contacts involved in hydrogen bonding.The Hirshfeld surfaces of compounds I, they reveal that the principal inter\u00admolecular contacts are H\u22efH , H\u22efC/C\u22efH , O\u22efH/H\u22efO , C\u22efC , S\u22efH/H\u22efS , S\u22efC/C\u22efS and N\u22efH/H\u22efN . For compound II, they reveal a similar trend, with the principal inter\u00admolecular contacts being H\u22efH/H\u22efH , H\u22efC/C\u22efH , O\u22efH/H\u22efO , C\u22efC , S\u22efH/H\u22efS , S\u22efC/C\u22efS , C\u22efO/O\u22efC and S\u22efO/O\u22efS . In both compounds, the H\u22efH inter\u00admolecular contacts predominate, followed by C\u22efH/H\u22efC and O\u22efH/H\u22efO contacts.The fingerprint plots are illustrated in Figs. 65.I: A solution of N-propargyl\u00adbenzene\u00adsulfonamide (0.50\u2005g) in xylenes (20\u2005mL), MnO2 (0.50\u2005g) was added and the reaction mixture was refluxed for 24\u2005h. It was then filtered through a celite pad and washed with hot xylenes (2 \u00d7 10\u2005mL). The combined filtrate was concentrated under vacuum and then triturated with MeOH to afford dibenzo[b]thio\u00adphene as a dull white solid. Finally, compound I was crystallized using ethanol.Compound II: To a solution of (E)-N-{2-[2-(benzo[b]thio\u00adphen-2-yl)ethenyl]phen\u00adyl}benzenesulfonamide in CH3CN (10\u2005mL), K2CO3 and propargyl bromide were added and the mixture was stirred at room temperature for 12\u2005h. After completion of the reaction (monitored by TLC), it was poured into crushed ice (50\u2005g) containing conc. HCl (5\u2005mL), extracted with ethyl acetate (2 \u00d7 20\u2005mL) then washed with water (2 \u00d7 20\u2005mL) and dried (Na2SO4). Removal of the solvent in vacuo followed by crystallization from methanol (4\u2005mL) afforded compound II as a white solid.Compound 6.Uiso(H) = 1.2Ueq(C) or 1.5Ueq(Cmeth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023003821/zn2028sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989023003821/zn2028Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989023003821/zn2028IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989023003821/zn2028Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023003821/zn2028IIsup5.cmlSupporting information file. DOI: 2259716, 2259715CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "We previously reported that diacylglycerol (DG) kinase (DGK) \u03b4 interacts with DG\u2010generating sphingomyelin synthase (SMS)\u2010related protein (SMSr), but not SMS1 or SMS2, via their sterile \u03b1 motif domains (SAMDs). However, it remains unclear whether other DGK isozymes interact with SMSs. Here, we found that DGK\u03b6, which does not contain SAMD, interacts with SMSr and SMS1, but not SMS2. Deletion mutant analyses demonstrated that SAMD in the N\u2010terminal cytosolic region of SMSr binds to the N\u2010terminal half catalytic domain of DGK\u03b6. However, the C\u2010terminal cytosolic region of SMS1 interacts with the catalytic domain of DGK\u03b6. Taken together, these results indicate that DGK\u03b6 associates with SMSr and SMS1 in different manners and suggest that they compose new DG signaling pathways. The sterile \u03b1 motif domain (SAMD) of sphingomyelin synthase (SMS)\u2010related protein (SMSr) binds to the C\u2010terminal half catalytic domain of diacylglycerol kinase (DGK) \u03b6. Moreover, the C\u2010terminal cytosolic region of SMS1 interacts with the catalytic domain of DGK\u03b6. SMSr\u2010SAMD associates with DGK\u03b4\u2010SAMD and activates DGK\u03b4. However, SMSr\u2010SAMD moderately suppresses the activity of DGK\u03b6 in contrast to DGK\u03b4. CPESceramide phosphoethanolamine synthaseDGdiacylglycerolDGKdiacylglycerol kinaseFLfull lengthHRPhorseradish peroxidasePAphosphatidic acidPAPphosphatidic acid phosphatasePCphosphatidylcholinePEphosphatidylethanolaminePIphosphatidylinositolPLCphospholipase CPSphosphatidylserineSAMDsterile \u03b1 motif domainSMSsphingomyelin synthaseSMSrsphingomyelin synthase\u2010related proteinTEVtobacco etch virusTMDtransmembrane domainTSTwin\u2010Strepl\u2010lactate dehydrogenase A, and creatine kinase muscle type [Diacylglycerol kinase (DGK) phosphorylates diacylglycerol (DG) to convert it to phosphatidic acid (PA) , 3, 4, 5cle type , 15, 16.Diacylglycerol kinase isozymes function in a wide variety of physiological events, including immunity, cell proliferation, and the central nervous system , 18. ForThere are three isoforms of sphingomyelin synthase (SMS) , 37, SMSWe recently searched for an upstream enzyme (a DG supply enzyme) of DGK\u03b4. Consequently, we found that DGK\u03b4 interacts with SMSr , which sIn the present study, we comprehensively searched for interactions between DGK isozymes and SMS isozymes. We found that DGK\u03b6 binds to SMS1 and SMSr but not SMS2. Moreover, DGK\u03b6 interacts with SMSr and SMS1 in different manners. These results suggest that, beyond our expectations, DGK isozymes and SMS isozymes form a complex network.Mouse monoclonal anti\u2010V5 antibody was obtained from Thermo Fisher Scientific . Rabbit polyclonal anti\u2010GFP antibody (#598) was purchased from Medical and Biological Laboratories . Mouse monoclonal anti\u2010FLAG\u2013tag antibody (F1804) was obtained from Sigma\u2013Aldrich . Mouse monoclonal anti\u2010GFP antibody (sc\u20109996) was purchased from Santa Cruz Biotechnology . Mouse monoclonal anti\u2010Strep II (M211\u20103) and rabbit polyclonal anti\u2010GST (PM013) were obtained from Medical and Biological Laboratories . A horseradish peroxidase (HRP)\u2010conjugated goat anti\u2010mouse IgG antibody was obtained from Bethyl Laboratories . Goat anti\u2010rabbit IgG\u2010HRP was purchased from Jackson ImmunoResearch .Plasmids for expressing N\u2010terminal 3\u00d7FLAG\u2010tagged human or rat DGK isozymes and for We used the following nomenclature for epitope\u2010tagged proteins: TagX\u2010(protein) and (protein)\u2010TagY means that TagX and TagY are located at the N and C termini of the protein, respectively.EcoRI/SalI sites. The amplicons were generated using the following primers: SMS1\u2010NT (aa 1\u2013134) using 5\u2032\u2010CTCAAGCTTCGAATTATGAAGGAAGTGGTTTATTG\u20103\u2032 (forward) and 5\u2032\u2010CCGCGGTACCGTCGATCACTTGCCCCACTCCATG\u20103\u2032 (reverse); SMS1\u2010CT (aa 348\u2013413) using 5\u2032\u2010CTCAAGCTTCGAATTCACACTATGGCCAATCAGC\u20103\u2032 (forward) and 5\u2032\u2010CCGCGGTACCGTCGATTATGTGTCATTCACCAGCC\u20103\u2032 (reverse); SMSr\u2010NT (aa 1\u2013151), 5\u2032\u2010CTCAAGCTTCGAATTATGGCAGGTCCTAATC\u20103\u2032 (forward) and 5\u2032\u2010CCGCGGTACCGTCGATCACTTCCAGTATTCTGGGTC\u20103\u2032 (reverse); SMSr\u2010CT (aa 364\u2013415), 5\u2032\u2010CTCAAGCTTCGAATTCATACTCTGGCCAATACC\u20103\u2032 (forward) and 5\u2032\u2010 CCGCGGTACCGTCGATCATCCAATTAGTCTTTTC\u20103\u2032 (reverse).The N\u2010terminal (NT) or C\u2010terminal (CT) cytosolic regions of SMS1 and SMSr were subcloned into the pAcGFP\u2010C1 vector via In\u2010Fusion cloning (Clontech\u2010Takara Bio) at the S\u2010transferase (GST)\u2010tagged SMS1\u2010CT and SMSr\u2010NT were generated by in\u2010fusion cloning. The pGEX\u20106P\u20101 vector was linearized at the EcoRI and SalI sites, and amplified gene with 15\u2009bp extensions homologous to vector ends. We generated SMS1\u2010CT using 5\u2032\u2010GGGATCCCCGGAATTCCACACTATGGCCAATCAGC\u20103\u2032 (forward) and 5\u2032\u2010GTCGACCCGGGAATTCTATGTGTCATTCACCAGCC\u20103\u2032 (reverse) and SMSr\u2010NT using 5\u2032\u2010GAATTCCCGGGTCGAATGGCAGGTCCTAATCAAC\u20103\u2032 (forward) and 5\u2032\u2010GGCCGCTCGAGTCGATCACTTCCAGTATTCTGGGTCC\u20103\u2032 (reverse).Glutathione The cDNAs encoding DGK isozymes that were subcloned into the expression plasmid, p3\u00d7FLAG\u2010CMV (Sigma\u2013Aldrich), for expression in mammalian cells were generated as described .EcoRI/SalI sites of the 3\u00d7FLAG CMV 7.1 vector. The following DGK\u03b6 mutants were generated using the following primers: DGK\u03b6\u2010NT (aa 1\u2013283), 5\u2032\u2010GGTGGTGAATTCAATGGAGCCGCGGGACGG\u20103\u2032 (forward) and 5\u2032\u2010ACGCGTCGACCTAGAAGGGTCTCCAGCGGCC\u20103\u2032 (reverse); DGK\u03b6\u2010CD (aa 284\u2013640), 5\u2032\u2010CCGGAATTCAATCATCAGGCCCACCCCC\u20103\u2032 (forward) and 5\u2032\u2010ACGCGTCGACCTACACCGGCTGCTGGTCG\u20103\u2032 (reverse); DGK\u03b6\u2010CD\u2010a (aa 284\u2013641), 5\u2032\u2010CCGGAATTCAATCATCAGGCCCACCCCC\u20103\u2032 (forward) and 5\u2032\u2010ACGCGTCGACCTACTCAGGCCCTGCCTCGG\u20103\u2032 (reverse); DGK\u03b6\u2010CD\u2010b (aa 433\u2013640), 5\u2032\u2010CCGGAATTCAGACCGAGATGAAGGCGCC\u20103\u2032 (forward) and 5\u2032\u2010ACGCGTCGACCTACACCGGCTGCTGGTCG\u20103\u2032 (reverse); DGK\u03b6\u2010CT (aa 641\u2013928) using the primers 5\u2032\u2010CCGGAATTCACCAGAGCAGTTGCGCATCC\u20103\u2032 (forward) and 5\u2032\u2010ACGCGTCGACCTACACAGCCGTCTCCTGGTC\u20103\u2032 (reverse).N\u2010terminal 3\u00d7FLAG\u2010tagged human DGK\u03b6 mutants were generated by PCR and inserted into the 2\u2010GGSA\u2010WSHPQFEK) was cloned into the XhoI/BglII site of the pCAGGS vector [BglII site of pCAGGS\u2010C\u2010TEV\u2010Twin\u2010Strep via In\u2010Fusion cloning. The following primers were used to amplify FL DGK\u03b6: forward, 5\u2032\u2010TTTTCAAGGCAGATCTATGGAGCCGCGGGACG\u20103\u2032; reverse, 5\u2032\u2010AGAGGGAAAAAGATCTCTACACAGCCGTCTCCTGG\u20103\u2032. N\u2010terminal Twin\u2010Strep\u2010tagged DGK\u03b6 was subcloned into pOET3 vector . The following primers were used, forward, 5\u2032\u2010TTTTCAAGGCAGATCTATGGAGCCGCGGGACG\u20103\u2032; reverse, 5\u2032\u2010TTATTAATTAAGATCTCTACACAGCCGTCTCCTGG\u20103\u2032.The plasmid expressing N\u2010terminal Tobacco Etch Virus (TEV) protease cleavable Twin\u2010Strep\u2010tag (ENLYFQGS\u2010WSHPQFEK\u2010(GGGS)S vector to gener\u22121 penicillin/100\u2009\u03bcg\u00b7mL\u22121 streptomycin at 37\u2009\u00b0C in an atmosphere containing 5% CO2. The plasmids were transiently transfected using PolyFect according to the manufacturer's instructions or using polyethylenimine Max [\u22121, pH 8.0) were preincubated for 10\u2009min at a 1\u2009:\u20093 ratio (20\u2009\u03bcg DNA: 60\u2009\u03bcL polyethylenimine Max) in 750\u2009\u03bcL of Opti\u2010MEM before transfection. The cells overexpressing recombinant proteins were harvested after 24\u2009h and the pellets were resuspended in 40% (v/v) glycerol diluted in phosphate\u2010buffered saline. The cell samples were flash\u2010frozen in liquid nitrogen and stored at \u221280\u2009\u00b0C until use.HEK293 cells were maintained in Dulbecco's modified Eagle's medium supplemented with 5% FBS (Thermo Fisher Scientific) and 100\u2009U\u00b7mLPA, USA) . The exp2 in the dark. Volume of the medium was kept at 20\u201330% of flask volume. To generate recombinant baculovirus was generated using pOET3 vector and the flashBAC system (Oxford Expression Technologies) as described previously [Sf9 cells were maintained in Sf\u2010900 II serum\u2010free medium in sterile Erlenmeyer flask at 120\u2009r.p.m. and 28\u2009\u00b0C without COeviously .HEK293 cell lysates expressing V5\u2010tagged SMS1, SMS2, SMSr, or their AcGFP\u2010tagged mutants and 3\u00d7FLAG\u2010tagged DGKs or their mutants were subjected to immunoprecipitation with anti\u2010V5 (MA5\u201015253) or anti\u2010GFP (#598) antibody and Protein A/G PLUS\u2010agarose beads (Santa Cruz Biotechnology) as described previously .S\u2010transferase\u2010fused SMS1\u2010CT and SMSr\u2010NT were bacterially expressed and highly purified using glutathione\u2010Sepharose beads . Twin\u2010Strep (TS)\u2010tagged DGK\u03b6 (TS\u2010DGK\u03b6) was expressed by mammalian cells and highly purified using Strep\u2010Tactin XT beads .Glutathione S\u2010transferase pull\u2010down assays were performed as previously [m Tris\u2013HCl, pH 7.5, 150\u2009mm NaCl, 1\u2009mm EDTA, 0.1% (v/v) Triton X\u2010100, and 1\u2009mm phenylmethylsulfonyl fluoride. Purified Twin\u2010strep tagged DGK\u03b6 was incubated with the beads for 2\u2009h at 4\u2009\u00b0C with constant rocking. Then, the beads were washed five times with buffer. The washed beads washed were then boiled in SDS sample buffer, and the extracts were analyzed by western blotting.Glutathione eviously . Purifiem Tris\u2013HCl, pH 7.4, containing 150\u2009mm NaCl, 10% (v/v) glycerol, 1\u2009mm PMSF, 0.1\u2009mm DTT, 20\u2009\u03bcg\u00b7mL\u22121 aprotinin, 20\u2009\u03bcg\u00b7mL\u22121 leupeptin, 20\u2009\u03bcg\u00b7mL\u22121 pepstatin, and 20\u2009\u03bcg\u00b7mL\u22121 soybean trypsin inhibitor) containing detergents (1% (w/v) n\u2010dodecyl\u2010\u03b2\u2010d\u2010maltoside (DDM) and 0.2% (w/v) cholesteryl hemisuccinate (CHS)). The supernatant (1% DDM soluble fraction) was isolated by ultracentrifugation at 100\u2009000\u2009g for 30\u2009min at 4\u2009\u00b0C and then purified using Strep\u2010Tactin XT beads. The beads were washed with the lysis buffer containing 0.05% DDM and 0.01% CHS. Subsequently, the bound proteins were eluted with the lysis buffer containing 0.05% DDM, 0.01% CHS, and 2.5\u2009mm d\u2010desthiobiotin (Sigma\u2013Aldrich). TS\u2010tagged DGK\u03b6 was purified using Strep\u2010Tactin XT beads without detergents.C\u2010terminally TS\u2010tagged human SMS1 and SMSr (SMS1\u2010TS and SMSr\u2010TS), and N\u2010terminally TS\u2010tagged DGK\u03b6 (TS\u2010DGK\u03b6) were expressed in HEK293 cells. SMS1\u2010TS and SMSr\u2010TS were lysed via lysed via homogenization on ice with ice\u2010cold lysis buffer (20\u2009mDiacylglycerol kinase activity was measured using liquid chromatography\u2013tandem mass spectrometry (LC\u2013MS/MS) as previously described .Western blotting was carried out as previously described . Equal qt test for the comparison of two groups or one\u2010way ANOVA followed by Tukey's or Dunnett's post hoc test for multiple comparisons using graphpad prism 8 to determine any significant differences. P\u2009<\u20090.05 was considered significant.Data are represented as the means\u2009\u00b1\u2009SDs and were analyzed by the Student's We first examined the interaction of all DGK isozymes with SMSr Fig.\u00a0. We confNext, we determined the interaction between 10 DGK isozymes and SMS1 Fig.\u00a0. As prevWe next examined the interaction between 10 DGK isozymes and SMS2 Fig.\u00a0. As prevWe next attempted to determine a DGK\u03b6\u2010interaction region in SMSr. AcGFP\u2010tagged N\u2010terminal (AcGFP\u2010SMSr\u2010NT) and C\u2010terminal (AcGFP\u2010SMSr\u2010CT) cytosolic regions of SMSr were generated Fig.\u00a0, and theA DGK\u03b6\u2010interaction region in SMS1 was also identified. For this purpose, AcGFP\u2010tagged N\u2010terminal (AcGFP\u2010SMS1\u2010NT) and C\u2010terminal (AcGFP\u2010SMS1\u2010CT) cytosolic regions of SMS1 were made Fig.\u00a0. InteresTo narrow the DGK\u03b6\u2010interaction area in the N\u2010terminal cytosolic region of SMSr, the DGK\u03b6\u2010interaction activity of SAMD alone of SMSr was tested. Figure To determine an SMSr\u2010interaction region in DGK\u03b6, we divided the protein into three parts, DGK\u03b6\u2010NT, DGK\u03b6\u2010CD, and DGK\u03b6\u2010CT Fig.\u00a0, and detNext, the interaction of DGK\u03b6\u2010NT, DGK\u03b6\u2010CD, and DGK\u03b6\u2010CT with SMS1\u2010CT was examined. Among them, 3\u00d7FLAG\u2010DGK\u03b6\u2010CD most strongly interacted with AcGFP\u2010SMS1\u2010CT Fig.\u00a0. AlthougWe further divided the catalytic domain of DGK\u03b6 into CD\u2010a and CD\u2010b Fig.\u00a0 and deteWe next examined whether DGK\u03b6 directly binds to SMSr and SMS1. GST\u2010fused SMSr\u2010NT and SMS1\u2010CT Fig.\u00a0 were bacin\u2009vitro [in\u2009vitro in the presence of 34:1 (16:0/18:1)\u2010DG. Intriguingly, SMSr, but not SMS1, moderately inhibited DGK\u03b6 activity in\u2009vitro . Moreoveity Fig.\u00a0. Moreoveity Fig.\u00a0 also attity Fig.\u00a0. These rIn the present study, we demonstrated for the first time that DGK\u03b6 interacts with SMS1 and SMSr but not SMS2 Figs\u00a0. DGK\u03b41 aWe previously reported that DGK\u03b4 associates with SMSr via the interaction between DGK\u03b4\u2010SAMD and SMSr\u2010SAMD . AlthougSMSr, but not SMS1, inhibited DGK\u03b6 activity Fig.\u00a0. TherefoIn the case of the SMSr\u2013DGK\u03b6 interaction, SAMD in the N\u2010terminal cytosolic region of SMSr binds to DGK\u03b6 Fig.\u00a0. In contSMSr\u2010SAMD interacted with DGK\u03b6\u2010CD\u2010a Fig.\u00a0. Howeverhttps://www.proteinatlas.org/ENSG00000149091\u2010DGKZ/tissue), SMSr (https://www.proteinatlas.org/ENSG00000156671\u2010SAMD8/tissue), and SMS1 (https://www.proteinatlas.org/ENSG00000198964\u2010SGMS1/tissue) are ubiquitously expressed in a variety of tissues [Although SMSr is a DG\u2010generating enzyme, its CPES activity is very low . We rece tissues . These rIn summary, in the present study, we demonstrated for the first time that DGK\u03b6 interacts with SMS1 and SMSr but not SMS2 Fig.\u00a0. DGK\u03b4 alThe authors declare no conflict of interest.https://www.webofscience.com/api/gateway/wos/peer\u2010review/10.1002/2211\u20105463.13628.The peer review history for this article is available at MF primarily designed and conducted the experiments and analyzed the data. CM, YN, and RS designed and conducted the experiments and analyzed the data. CM, FS, and MF wrote the manuscript. CM and FS conceived the research. All authors revised the manuscript and approved its final version.Fig. S1. Sequence alignment of SMSr\u2010SAMD and SMS1\u2010SAMD. (A) Sequence alignment of SMSr\u2010SAMD (aa 12\u201378) and SMS1\u2010SAMD (aa 7\u201370). Sequence alignment was created using Clustal Omega provided by EMBL's European Bioinformatics Institute (EMBL\u2010EBI). Compared with SMSr\u2010SAMD, white letters on a black background indicate fully conserved residues, and black letters on a gray background indicate strongly similar residues. (B) Amino acid identities between the SAMDs of SMSr and SMS1. Amino acid identity and similarity were determined using Pairwise Sequence Alignment provided by the European Molecular Biology Open Software Suite (EMBOSS).Fig. S2. Multiple sequence alignment of the C\u2010terminal regions of SMS1, SMS2, and SMSr. (A) Multiple sequence alignment of the C\u2010terminal regions of SMS1\u2010CT (aa 348\u2013413), SMS2\u2010CT (aa 292\u2013365), and SMSr\u2010CT (aa 364\u2013415). Multiple sequence alignment was created using Clustal Omega provided by EMBL's European Bioinformatics Institute (EMBLEBI). Compared with SMS1\u2010CT, white letters on a black background indicate fully conserved residues, and black letters on a gray background indicate strongly similar residues. (B) Amino acid identities between the C\u2010terminal regions of SMS1, SMS2, and SMSr. Amino acid identity and similarity were determined using Pairwise Sequence Alignment provided by the European Molecular Biology Open Software Suite (EMBOSS).Fig. S3. Sequence alignment of DGK\u03b6\u2010CD and DGK\u03b9\u2010CD. (A) Sequence alignment of DGK\u03b6\u2010CD (aa 293\u2013622) and DGK\u03b9\u2010CD (aa 374\u2013702). Sequence alignment was created using Clustal Omega provided by EMBL's European Bioinformatics Institute (EMBL\u2010EBI). Compared with DGK\u03b6\u2010CD, white letters on a black background indicate fully conserved residues, and black letters on a gray background indicate strongly similar residues. (B) Amino acid identities between DGK\u03b6\u2010CD and DGK\u03b9\u2010CD. Amino acid identity and similarity were determined using Pairwise Sequence Alignment provided by the European Molecular Biology Open Software Suite (EMBOSS).Click here for additional data file."} +{"text": "The crystal structure of bucetin, an analgesic and anti\u00adpyric similar to phenacetin, is presented. N-(4-eth\u00adoxy\u00adphen\u00adyl)-3-hydroxy\u00adbutanamide], C12H17NO3, the mol\u00adecule is in an extended conformation as illustrated by the C\u2014O\u2014C\u2014C torsion angle [170.14\u2005(15)\u00b0] in the eth\u00adoxy group and the subsequent C\u2014N\u2014C\u2014C [\u2212177.24\u2005(16)\u00b0], N\u2014C\u2014C\u2014C [170.08\u2005(15)\u00b0] and C\u2014C\u2014C\u2014C [171.41\u2005(15)\u00b0] torsion angles in the butanamide chain. In the crystal, the O\u2014H group donates an inter\u00admolecular O\u2014H\u22efO hydrogen bond to the amide carbonyl oxygen atom and also accepts an inter\u00admolecular N\u2014H\u22efO hydrogen bond from an adjacent N\u2014H group. The former forms 12-membered dimeric rings about inversion centers, and the latter form chains in the [001] direction. The overall hydrogen-bonded network is two-dimensional, with no propagation in the [100] direction.In the title compound, racemic bucetin [systematic name: N-(4-Eth\u00adoxy\u00adphen\u00adyl)-3-hydro\u00adbutanamide, popularly known as bucetin, is an analgesic and anti\u00adpyric that is similar in structure to phenacetin [N-(4-eth\u00adoxy\u00adphen\u00adyl)acetamide]. Once thought to be a better substitute for phenacetin hydroxyl\u00adamine and 1-eth\u00adoxy-4-nitroso\u00adbenzene) on PGE2 synthesis and a possible reduction of COX-2 expression acetamide and 2.8611\u2005(19)\u2005\u00c5 for N\u2014H\u22efO was obtained from Sigma-Aldrich, St. Louis, MO and was used without further purification. Single crystals of racemic bucetin were prepared by slow cooling of a nearly saturated solution of bucetin in boiling deionized water.The title compound, CCrystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314623002316/vm4059sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314623002316/vm4059Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623002316/vm4059Isup3.cmlSupporting information file. DOI: 2247342CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure, the Th atom (site symmetry 3\u221e[ThBr4/2Br4/2]. In comparison with the previous crystal structure refinement , the current rerefinement resulted in much higher preciscion of the lattice parameters and the atomic coordinates.Single crystals of \u03b1-ThBr al. 1974. J. Less A comparison of the structural parameters of the original crystal structure refinement and of the current rerefinement is given in Table\u00a01The crystal structure of \u03b1-ThBra, site symmetry f, site symmetry 1) in the asymmetric unit. The Th atom is surrounded by eight Br atoms to form a tetra\u00adgonal-disphenoidal coordination polyhedron. The Th\u2014Br bond lengths of 4\u00a0\u00d7 2.9100\u2005(4)\u2005\u00c5 and 4\u00a0\u00d7 3.0107\u2005(4)\u2005\u00c5 are in good agreement with previously reported values of 2.909 and 3.020\u2005\u00c5 , with values of 2.85 and 3.12\u2005\u00c5 \u2005\u00c5 in \u03b1-ThBr4 is shorter compared to \u03b2-ThBr4, with a value of 4.8774\u2005\u00c5 \u2005\u00c5 and 4\u00a0\u00d7 6.70680\u2005(19)\u2005\u00c5.Fig.\u00a01\u22123\u2005mbar; 1\u2005bar = 105\u2005Pa) at least three times before use. Aluminium bromide was sublimed in vacuo before use; \u03b2-ThBr4 was prepared according to a literature protocol using a fine-vacuum line and a glove-box (MBraun). Silica ampoules were flame-dried under dynamic fine vacuum and CuBr , and sealed under vacuum. The ampoule was heated in a furnace to 753\u2005K at a rate of 1\u2005K\u2005min\u22121 and kept at this temperature for 480\u2005h for the reaction to take place. Afterwards, it was cooled to 330\u2005K at a rate of 50\u2005K\u2005d\u22121. Several colourless crystals of \u03b1-ThBr4 were obtained.A silica glass ampoule was loaded with \u03b2-ThBrCrystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314623008908/wm4200sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2414314623008908/wm4200Isup2.hklStructure factors: contains datablock(s) I. DOI: 2300477CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The carbamate group has key bond lengths of 1.404\u2005(6)\u2005\u00c5 (N\u2014C), 1.330\u2005(5)\u2005\u00c5 (O\u2014C), and 1.201\u2005(6)\u2005\u00c5 (C=O). The crystal contains inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions, as well as both type I and type II inter\u00admolecular I\u22efI inter\u00adactions.The mol\u00adecular structure of These inter\u00adactions have Cg\u22efCg distances of 3.484\u2005(3) and 3.589\u2005(3)\u2005\u00c5 with slippages of 1.028 and 1.376\u2005\u00c5 and angles of 0.00\u2005(3)\u00b0 . The supra\u00admolecular pillars are held together via both type I and type II inter\u00admolecular I\u22efI inter\u00adactions with an angle of 147.68\u2005(13)\u00b0 and an I\u22efI distance of 3.6630\u2005(5)\u2005\u00c5. The type II inter\u00adaction is found between atoms C11\u2014I2\u22efI1 with an I\u22efI distance of 3.8332\u2005(5)\u2005\u00c5 and an angle of 46.69\u2005(13)\u00b0.In the crystal, mol\u00adecules of the title compound form pillars 4.et al., 2016et al., 2015I with iodine atoms at the C5 and C11 positions, but where the nitro\u00adgen atom has been alkyl\u00adated with either a butyl or benzyl group. Structure ECUNUM bears bromine atoms at the C5 and C11 positions with a phenyl\u00adcarbamate group on the nitro\u00adgen atom = 1.2Ueq(C) for aromatic hydrogen atoms and Uiso(H) = 1.5Ueq(C) for the hydrogen atoms of the methyl group.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S205698902300230X/wm5676sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902300230X/wm5676Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902300230X/wm5676Isup3.cmlSupporting information file. DOI: 2247288CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-chromen-4-yl\u00adamino)\u00adbenzoate, has been synthesized and characterized.A new coumarin derivative, methyl 2-(2-oxo-2 H-chromen-4-yl\u00adamino)\u00adbenzoate, C17H13NO4 (1), was pre\u00adpared by condensation between 4-hy\u00addroxy\u00adcoumarin and methyl 2-amino\u00adbenzoate. It crystallizes in the ortho\u00adrhom\u00adbic space group Pca21 at 300\u2005K. The mol\u00adecule of compound 1 consists of the 2H-chromen-2-one part connected by an amine moiety (\u2013NH\u2013) to the methyl benzoate ring. The supra\u00admolecular array is formed by hydrogen bonds between the aromatic ring and the O atoms of the lactone and ester portions. The structural details match the spectroscopic data acquired from NMR and IR spectroscopy.Methyl 2-(2-oxo-2 These sDipteryx odorata Wild; Fabaceae family) by Vogel in 1820. Since then, more than 1300 coumarins have been identified from natural sources \u00adbenzoate, 1 \u00adbenzoate mol\u00adecule , although the C4\u2014N1\u2014C9 angle at 130.9\u2005(4)\u00b0 is about 7\u00b0 larger than in those mol\u00adecules, presumably due to the intra\u00admolecular N1\u2014H1\u22efO2 hydrogen bond (Table\u00a01The average C\u2014C bond distance in the aromatic portion of the coumarin is 1.374\u2005(7)\u2005\u00c5, while the C9\u2014C13, C9\u2014C10 and C10\u2014C11 bond lengths in the lactone portion are 1.450\u2005(7), 1.353\u2005(7) and 1.412\u2005(7)\u2005\u00c5, respectively, because of the partial localization of \u03c0-bonding within the ring. The C11\u2014O3 and C12\u2014O3 bond lengths are equivalent at 1.374\u2005(7) and 1.373\u2005(7)\u2005\u00c5, respectively, while the C11=O4 distance is 1.204\u2005(7)\u2005\u00c5. The sum of the angles about N1 is 359\u2005(3)\u00b0, implicating involvement of its lone pair in N\u2014C \u03c0-bonding. This is supported by the N1\u2014C9 and N1\u2014C4 distances of 1.351\u2005(6) and 1.391\u2005(6)\u2005\u00c5, respectively. Similar geometrical parameters are found in closely related structures \u00b0.1H and 13C NMR confirms the product as methyl 2-(2-oxo-2H-chromen-4-yl\u00adamino)\u00adbenzoate. In the 1H NMR spectrum, there is a singlet at \u03b4 3.74 ppm attributable to the meth\u00adoxy group of the ester, the coumarin vinylic H atom appears at \u03b4 5.31 ppm and a singlet is seen at \u03b4 9.67 which can be assigned to N\u2014H. In addition, there are eight aromatic H atoms between \u03b4 7.44 and 8.14 ppm. In the 13C NMR spectrum, the meth\u00adoxy group appears at \u03b4 52.47 ppm, the two carbonyl C atoms at \u03b4 166.50 and 161.40, and the vinylic and aromatic C atoms between \u03b4 114 and 154 ppm.The NMR spectra are shown in Figs. 43.B\u22efO4i) and the H atom from the aromatic ring (C7\u2014H7\u22efO4ii) Table\u00a01. These bon Fig.\u00a06. The cryon Fig.\u00a06 involves4.et al., 2016H-chromen-2-one -2H-chromen-2-one was heated in a 50\u2005ml Becher at 453\u2005K for 1\u2005h. A solution comprised of 30\u2005ml of hot methanol and 30\u2005ml of aqueous NaOH (1\u2005mol\u2005l\u22121) was then added to the solid. This mixture was stirred for 30\u2005min at 333\u2005K and then filtered. The solid was washed with water, dried and used without further purification.The reaction was carried out according to the literature I. DOI: 10.1107/S2056989023007351/mw2198Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023007351/mw2198Isup3.cmlSupporting information file. DOI: 2289922CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure features C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds, which link the mol\u00adecules into layers parallel to the (100) plane. IC\u2014H\u22ef\u03c0 inter\u00adactions and weak van der Waals inter\u00adactions occur between the layers. 25H17N3O5S2, intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid-to-centroid distance = 3.5640\u2005(9)\u2005\u00c5] are observed between the furan and benzene rings of the 4-cyano\u00adphenyl group. In the crystal, mol\u00adecules are connected via C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds, forming layers parallel to the (100) plane. These layers are inter\u00adconnected by C\u2014H\u22ef\u03c0 inter\u00adactions and weak van der Waals inter\u00adactions. Hirshfeld surface analysis indicates that H\u22efH (30.2%), N\u22efH/H\u22efN (22.3%), C\u22efH/H\u22efC (17.9%) and O\u22efH/H\u22efO (15.4%) inter\u00adactions make the most significant contributions to the crystal packing.In the title compound, C Reactions with ammonia, and primary and secondary amines are the most widespread. A primary amine will form a soluble sulfonamide salt in the presence of aqueous alkali (either KOH or NaOH). A secondary amine in the same reaction forms an insoluble sulfonamide. The most widely used sulfonyl\u00adamide is sulfanil\u00adamide, an anti\u00adbacterial drug that was first obtained in 1908 by the Austrian chemist Paul Josef Jakob Gelmo while he was trying to synthesize a dye for textile materials .In the title compound Fig.\u00a02, the angCg1\u22efCg4 = 3.5640\u2005(9)\u2005\u00c5; Cg1 and Cg4 are the centroids of the furan (O1/C7\u2013C10) and benzene rings (C19\u2013C24), respectively, of one of the two 4-cyano\u00adphen\u00adyl)sulfonyl groups, respectively; slippage = 0.793\u2005\u00c5], ensures the stability of the mol\u00adecular configuration.Intra\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions between the furan and a phenyl ring of one of the two (3-nitro\u00adphen\u00adyl)sulfonyl groups result in chains parallel to the a axis. In JOBTIF (space group P21/n), mol\u00adecules are linked by pairs of C\u2014H\u22efO hydrogen bonds, forming inversion dimers. In CEGMIM (space group Pbca), mol\u00adecules are connected by C\u2014H\u22efO inter\u00adactions into sheets in the ab plane. In the crystal of CEGSUE , mol\u00adecules associate via N\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds, forming extended hydrogen-bonded sheets that lie parallel to the bc plane. The N\u2014H\u22efN hydrogen bonds propagate along the b-axis direction, while the N\u2014H\u22efO hydrogen bonds propagate along the c-axis direction. The crystal structure of OCABUR (space group P21/c) features C\u2014H\u22efO hydrogen bonds. In the crystal structure of AYUPUG (space group P21/c), weak C\u2014H\u22efO inter\u00adactions connect the mol\u00adecules in a zigzag manner along the a-axis direction. In the crystal of PONZIC (space group Pa axis by inter\u00admolecular C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions. In ROGJON (space group Pbca), the crystal structure features weak inter\u00admolecular N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions.In PIMGUR was added gradually to a solution of 2-(5-methyl-2-fur\u00adyl)aniline in pyridine (7\u2005mL) under stirring and cooling in an ice\u2013water bath. The mixture was stirred for 7\u2005h at r.t. and after completion of the reaction , the mixture was poured into hydro\u00adchloric acid . The separated oil was washed with water until its crystallization. The obtained crystals were filtered off, dried, and recrystallized from an ethanol/di\u00admethyl\u00adformamide (DMF) mixture to give the target disulfonamide as a colourless solid. Single crystals were obtained by slow crystallization from an EtOH/DMF mixture . IR (KBr), \u03bd (cm\u22121): 1156 (\u03bds SO2), 1329 (\u03bdas SO2), 2237 (CN). 1H NMR : \u03b4 8.08 , 7.90 , 7.72 , 7.56 , 7.36 , 7.04 , 6.61 , 5.93 , 1.96 ; 13C{1H} NMR : \u03b4 153.2, 147.9 (2C), 142.9, 134.0 (4C), 133.7, 132.2, 132.0, 129.6 (4C), 128.8, 128.7, 128.6, 117.9 (2C), 117.4 (2C), 112.0, 108.6, 13.4; MS (ESI) m/z: [M + H]+ 504. Elemental analysis calculated (%) for C25H17N3O5S2 %: C 59.63, H 3.40, N 8.34, S 12.74; found: C 60.00, H 3.27, N 8.56, S 13.03.6.Ueq(C) (1.5 for methyl groups). The hydrogen atoms of the methyl group containing the C11 atom were disordered over two positions with equal occupancies.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023006254/tx2071sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023006254/tx2071Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023006254/tx2071Isup3.cmlSupporting information file. DOI: 2282070CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, N\u2014H\u22efO and C\u2014H\u22efO hydro\u00adgen bonds form chains extending along the b-axis direction. Due to the disorder of the p-tolyl rings, short C\u22efC distances are observed between adjacent chains.The quinoxaline moiety in the title mol\u00adecule, C Lgaz et al., 2015e.g. Abad et al., 2021Among the various classes of nitro\u00adgen heterocyclic compounds, quinoxaline derivatives display a broad spectrum of biological and pharmacological activities \u00b0 apart in a 0.503\u2005(2):0.497\u2005(2) ratio. In the crystal, N3\u2014H3A\u22efO2 and C10\u2014H10B\u22efO2 hydro\u00adgen bonds (Table\u00a01b-axis direction. Pairs of inversion-related chains show C16\u22efC17i and C17\u22efC16i distances of 2.695\u2005(4)\u2005\u00c5, which is 0.71\u2005\u00c5 less than the sum of the van der Waals radii and is likely due to the disorder involving this ring. The chains stack along the c-axis direction \u00b0 between the mean planes through the constituent rings. The s Table\u00a01 form chan Figs.\u00a02 and\u00a03 \u25b8.H)-one was dissolved in 25\u2005ml of di\u00admethyl\u00adformamide and 1.15\u2005g (6.24\u2005mmol) of 2-chloro-N-(p-tol\u00adyl)acetamide were added, followed by 1.0\u2005g (7.5\u2005mmol) of potassium bicarbonate, and a spatula tip of BTBA (benzyl\u00adtri\u00adbutyl\u00adammonium chloride) was used as a phase-transfer catalyst. The reaction was stirred for 2\u2005h under reflux at 353\u2005K. When the starting reagents had completely reacted, 500\u2005ml of distilled water were added and a few minutes later the product precipitated. This was filtered off, dried and recrystallized from hot ethanol solution to yield light-yellow plate-like crystals of the title compound.1.00\u2005g (6.24\u2005mmol) of 3-methyl\u00adquinoxalin-2(1I/\u03c3(I) > 20 and chosen from the full data set with CELL_NOW :0.497\u2005(2) ratio] of the disordered C12\u2013C17 ring were refined as rigid hexa\u00adgons.Crystal, data collection and refinement details are presented in Table\u00a0210.1107/S2414314623003577/vm4060sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314623003577/vm4060Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623003577/vm4060Isup3.cmlSupporting information file. DOI: 2254194CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Correction: BMC Ophthalmol\u00a023, 279 (2023)https://doi.org/10.1186/s12886\u2013023-03012-1After publication of the original article , the autThe original article has been updated and the correct values are also given below in bold.The second paragraph in page 4 should read:28.6\u00a0mg/dL, and apolipoprotein A1\u00a0131.6\u00a0mg/dL. A total of 782 patients in the alirocumab group had\u2009\u2265\u20092 consecutive LDL-C values\u2009<\u200915\u00a0mg/dL (0.39\u00a0mmol/L) and were matched to 2346 patients from the placebo group with similar baseline characteristics (Tables 4 and 5). Baseline characteristics of these patients included mean age 59\u00a0years, male sex (81%), diabetes (33%); and mean body mass index 27.3\u00a0kg/m2, LDL-C 2.0\u00a0mmol/L, lipoprotein (a) 18.8\u00a0mg/dL, and apolipoprotein A1\u00a0129.9\u00a0mg/dL.A total of 4305 patients in the alirocumab group had\u2009\u2265\u20092 consecutive LDL-C values\u2009<\u200925\u00a0mg/dL (0.65\u00a0mmol/L) and were matched to 4305 patients from the placebo group with similar baseline characteristics (Tables 2 and 3). Baseline characteristics of these patients included mean age 59\u00a0years, male sex (81%), diabetes (33%); and mean body mass index 28.3\u00a0kg/m2, LDL-C 2.1\u00a0mmol/L, lipoprotein (a)"} +{"text": "The layers are connected by additional C\u2014H\u22efO hydrogen bonds and \u03c0-stacking inter\u00adactions.In the title compound, the thia\u00adzine ring exhibits a screw-boat conformation. In the crystal, corrugated layers of mol\u00adecules parallel to the 8H7NO3S, the nitro\u00adgen atom has a planar environment, and the thia\u00adzine ring exhibits a screw-boat conformation. In the crystal, corrugated layers of mol\u00adecules parallel to the ab plane are formed by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds together with C\u2014H\u22ef\u03c0(ring) and S=O\u22ef\u03c0(ring) inter\u00adactions. The layers are connected by additional C\u2014H\u22efO hydrogen bonds and \u03c0-stacking inter\u00adactions. Hirshfeld surface analysis indicates that the most important contributions for the crystal packing are from H\u22efO/O\u22efH (49.4%), H\u22efH (23.0%) and H\u22efC/C\u22efH (14.1%) inter\u00adactions. The volume of the crystal voids and the percentage of free space were calculated as 75.4\u2005\u00c53 and 9.3%. Density functional theory (DFT) computations revealed N\u2014H\u22efO and C\u2014H\u22efO hydrogen-bonding energies of 43.3, 34.7 and 34.4\u2005kJ\u2005mol\u22121, respectively. Evaluation of the electrostatic, dispersion and total energy frameworks indicate that the stabilization is dominated via the electrostatic energy contribution. Moreover, the DFT-optimized structure at the B3LYP/ 6\u2013311\u2005G level is compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.In the title mol\u00adecule, C The distorted screw-boat conformation places O2 in an axial position and O3 in a pseudo-equatorial position \u2005\u00c5, S1\u22efCg2 = 4.1655\u2005(5)\u2005\u00c5, S1=O2\u22efCg2i = 101.77\u2005(3)\u00b0; symmetry code: (i) \u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a0Cg2\u22efCg2 = 3.7353\u2005(5)\u2005\u00c5, slippage = 1.55\u2005\u00c5] into a tri-periodic network structure with 4.9% contribution to the HS is viewed at de + di = 3.30\u2005\u00c5. The C\u22efC contacts appearing as a bullet-shaped distribution of points make a contribution of 3.7% to the HS with the tip at de = di = 1.70\u2005\u00c5. The spikes of H\u22efN/N\u22efH contacts with 3.2% contribution to the HS are viewed at de + di = 2.75\u2005\u00c5. Finally, the O\u22efO and C\u22efN/N\u22efC contacts contribute 1.3% and 0.4%, respectively, to the HS. The Hirshfeld surface representations with the function dnorm plotted onto the surface are shown for the H\u22efO/O\u22efH, H\u22efH and H\u22efC/C\u22efH inter\u00adactions in Fig.\u00a08a\u2013c, respectively. The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efO/O\u22efH, H\u22efH and H\u22efC/C\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions play the major role in the crystal packing u. Fig.\u00a04, the whits Fig.\u00a07e with 4ts Fig.\u00a07f appearts Fig.\u00a07g with 3\u22efO Fig.\u00a07h and C\u22ef\u22efC Fig.\u00a07i contacet al., 2011a,b) and the percentage of free space in the unit cell are calculated as 75.4\u2005\u00c53 and 9.3%, respectively. Thus, the crystal packing appears compact and the mechanical stability should be substantial.The strength of the crystal packing is important for determining the response to an applied mechanical force. If the crystal packing results in significant voids, then the mol\u00adecules are not tightly packed and a small amount of applied external mechanical force may easily break the crystal. To check the mechanical stability of the crystal, a void analysis was performed by adding up the electron densities of the spherically symmetric atoms contained in the asymmetric unit is the sum of electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies were calculated to be for N1\u2014H1\u22efO3, for C8\u2014H8B\u22efO2 and for C5\u2014H5\u22efO1.The inter\u00admolecular inter\u00adaction energies were calculated using the CEB3LYP/631G energy model available in et al., 2015Eele Energy frameworks combine the calculation of inter\u00admolecular inter\u00adaction energies with a graphical representation of their magnitude EHOMO and ELUMO, electronegativity (\u03c7), hardness (\u03b7), dipole moment (\u03bc), electrophilicity (\u03c9) and softness (\u03c3) are compiled in Table\u00a03\u03b7 and \u03c3 are essential for assessing reactivity and stability. The electron transition from HOMO to LUMO energy levels is depicted in Fig.\u00a011b]thia\u00adzin-3(4H)-one 1,1-dioxide ring. The energy band gap [\u0394E = ELUMO\u00a0\u2212\u00a0EHOMO] for the mol\u00adecule is 11.7261\u2005eV, and the energies of the frontier mol\u00adecular orbitals, EHOMO and ELUMO, are \u22129.6740\u2005eV and 2.0522\u2005eV, respectively.The highest-occupied mol\u00adecular orbital (HOMO), functioning as an electron donor, and the lowest-unoccupied mol\u00adecular orbital (LUMO), acting as an electron acceptor, serve as vital parameters in quantum chemistry. A small energy gap signifies high mol\u00adecular polarizability and enhanced chemical reactivity. The DFT calculations provided crucial insights into the reactivity and site selectivity of the mol\u00adecular framework. Parameters such as 7.et al., 2016II was dissolved in 3\u2005ml of acetic acid and added dropwise into a solution of potassium permanganate (1.81\u2005mmol) in 6\u2005ml of water. After stirring for one\u2005h at room temperature, a solution of sodium thio\u00adsulfate penta\u00adhydrate (20%wt) was added to react with excessive potassium permanganate. The precipitate obtained was filtered and recrystallized from ethanol to yield single-crystals suitable for X-ray structure analysis..3,4-Di\u00adhydro-29.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S205698902300868X/wm5698sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S205698902300868X/wm5698Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902300868X/wm5698Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S205698902300868X/wm5698Isup4.cmlSupporting information file. DOI: 2298958CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The various inter\u00admolecular inter\u00adactions, such as N\u2014H\u22efO, C\u2014H\u22efN and C\u2014H\u22efO, were investigated using Hirshfeld surface analysis and the three-dimensional inter\u00adaction energies were calculated. 29H27F2N3O6, which crystallizes in the monoclinic space group P21/c, the cyclo\u00adhexenone ring is puckered and adopts an envelope conformation. The crystal structure features various inter\u00admolecular inter\u00adactions, such as N\u2014H\u22efO, C\u2014H\u22efN and C\u2014H\u22efO. These inter\u00adactions were investigated using Hirshfeld surface analysis and the three-dimensional inter\u00adaction energies were calculated using the B3LYP/6\u201331\u2005G energy density model.In the title compound, C The orientation of other two carboxyl\u00adate groups at the C4 and C5 positions are described by the torsion angles C1\u2014C6\u2014C27\u2014O6 = \u221215.4\u2005(5)\u00b0 (\u2013syn-periplanar), C1\u2014C6\u2014C27\u2014O5 = 167.2\u2005(4)\u00b0 (+anti-periplanar), C5\u2014C6\u2014C27\u2014O5 = \u221217.2\u2005(6)\u00b0 (\u2013anti-periplanar), C5\u2014C6\u2014C27\u2014O6 = 160.1\u2005(3)\u00b0 (+anti-periplanar) and C3\u2014C4\u2014C23\u2014O3 = 44.9\u2005(5)\u00b0 , C3\u2014C4\u2014C23\u2014O4 = \u2212136.4\u2005(3)\u00b0 , C5\u2014C4\u2014C23\u2014O3 = \u221275.8\u2005(4)\u00b0 , C5\u2014C4\u2014C23\u2014O4 = 102.9\u2005(3)\u00b0 . The orientation is due to the inter\u00admolecular N\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions.In the title compound Fig.\u00a01, the cyc3.N\u22efO1, C14\u2014H14B\u22efN3, and C24\u2014H24B\u22efO3 propane-1,2,3-tri\u00adcarboxyl\u00adate was computed using two-dimensional fingerprint calculations mapped over ns Fig.\u00a05. The res6.CrystalExplorer 17.5 software calculates inter\u00adaction energies between crystal mol\u00adecular pairs. Energy calculations were carried out using the B3LYP/6-31G basis set within a default radius of 3.8\u2005\u00c5 is associated with a pair of yellow mol\u00adecules with the short centroid distance R = 9.29\u2005\u00c5 with rotational symmetry \u2212x, y\u00a0+\u00a0z\u00a0+\u00a0Etot = \u221217.6\u2005kJ\u2005mol\u22121) was observed for a pair of green mol\u00adecules inter\u00adacting at the longer centroid distance R = 12.86\u2005\u00c5; this is in accordance with the classical laws of electrostatics. In each of the energy terms, the dispersion component is dominant over the others.7.2SO4, then concentrated under reduced pressure to get the crude product. This was purified by silica gel column chromatography using n-hepta\u00adne/ethyl acetate as eluent. The mixture was quenched in cold water and the organic layer was extracted with ethyl acetate, washed with 5% sodium bicarbonate solution, and dried over anhydrous sodium sulfate. Slow evaporation of the solvent lead to crystals of the title compound, which were recrystallized from ethanol solution.Piperidine (6\u2005mmol) was added to ethyl cyano\u00adacetate (30\u2005mmol) and the mixture was stirred for 10\u2005min. Then 4-fluoro\u00adbenzaldehyde (20\u2005mmol) was added dropwise and during the addition, the temperature of the reaction mass rose to 333\u2005K (it should not be cooled), and the mass was stirred for 30\u2005min. The temperature slowly came down to 293\u2013298\u2005K over 30\u2005min. The progress of the reaction was monitored by TLC and found to be complete. Methyl\u00adene chloride (30\u2005ml) and water (20\u2005ml) were added and the mixture was stirred for 10\u2005min. The organic layer was separated and washed with sat. aq. NaCl solution and dried over anhydrous Na8.Uiso(H) = 1.2Ueq(C) (1.5 for methyl H atoms).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023003134/ex2065sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989023003134/ex2065Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023003134/ex2065Isup3.cmlSupporting information file. DOI: 2202315CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "However, it remains largely unclear how those cells develop. Here we show that transforming growth factor beta (TGF-\u03b2) signaling controls the development of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs. Deletion of TGF-\u03b2 receptors or Smad3 and Smad2 in bone marrow stem cells caused a deficiency of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs in mixed bone marrow chimeric mice. Mechanistically, TGF-\u03b2 is required for the development of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs thymic precursors (CD44\u2013CD25\u2013 \u03b3\u03b4 thymocytes). In addition, TGF-\u03b2 signaling induced CD8\u03b1 in thymic \u03b3\u03b4T cells and maintained CD8\u03b1 expression and survival in TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs. Moreover, TGF-\u03b2 also indirectly controls TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs by modulating the function of intestinal epithelial cells (IECs). Importantly, TGF-\u03b2 signaling in TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs safeguarded the integrity of the intestinal barrier in dextran sulfate sodium (DSS)-induced colitis.\u03b3\u03b4 intestinal intraepithelial lymphocytes (IELs) constitute the majority of IELs with unique CD8\u03b1\u03b1 T cell receptor (TCR)+ IELs, which express either TCR\u03b3\u03b4+ or TCR\u03b1\u03b2+ are usually classified into conventional and unconventional IELs according to their distinct phenotype and developmental pathways. \u03b3\u03b4 IELs are thought to be homed to the intestine immediately after their generation from \u03b3\u03b4T precursors in the thymus2. They comprise the vast majority of IELs in the small intestine and are essential to maintaining immune homeostasis in the intestinal territory, such as keeping the integrity of the gut barrier, limiting translocation of microbiomes, responding to antigens invasion, and healing tissue damage. Accumulated studies have shown that the function of \u03b3\u03b4 IELs is tightly related to their crosstalk with IECs; the dynamic movement of \u03b3\u03b4 IELs surrounding IECs results in more efficient immune surveillance and higher expression of antimicrobial or antiviral genes5.Intestinal intraepithelial lymphocytes (IELs) localize within the intestinal epithelium and predominate in the mucosal immune system; they are typically surrounded by intestinal epithelial cells (IECs) with a ratio of ~1:10 (IELs:IECs) in the small intestine+ homodimers that are considered the main cell resource for the production of cytokines like IFN-\u03b3, IL-10, and IL-13 from \u03b3\u03b4 IELs1. However, it remains uncertain which factors determine the development of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs. The development of \u03b3\u03b4T cells in the thymus undergoes several stages from DN1 to DN4 that can be classified by CD44 and CD25 expression, namely CD44+CD25\u2013 (DN1), CD44+CD25+ (DN2), CD44\u2013CD25+ (DN3), and CD44\u2013CD25\u2013 (DN4)8. However, it has been recently suggested that the V\u03b37+ subset of IELs were generated in an extrathymic way, which relies on Butyrophilin-like (Btnl) molecules on IECs9, but this remains to be verified.Unlike systemic \u03b3\u03b4T cells settled in other sites, roughly 90% of \u03b3\u03b4 IELs have a specific phenotype of CD8\u03b1\u03b112. TGF-\u03b2 is enriched in the intestinal environment and represents one of the most important regulators in gut immune system as both IECs and immune cells, including IELs, contribute to TGF-\u03b2 production14. Additionally, we and others have previously found that TGF-\u03b2 signaling is crucial for the development of TCR\u03b1\u03b2+CD8\u03b1\u03b1+ IELs and the generation of TCR\u03b1\u03b2+CD8\u03b1+CD4+ IELs16. However, it remains unknown whether TGF-\u03b2 plays a role in the development of TCR\u03b3\u03b4+ CD8\u03b1\u03b1+ IELs.TGF-\u03b2 signaling is involved in the development of various immune cells in the thymus and periphery, such as \u03b1\u03b2T cells, T regulatory cells (Tregs), and Th17 cells+CD8\u03b1\u03b1+ IELs in a Smad2 and Smad3 (Smad2/3)-dependent manner. Mice lacking TGF-\u03b2 receptors or Smad2/3 have fewer TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs and thymic \u03b3\u03b4 precursors capable of migrating to the intestine. We discovered that TGF-\u03b2 induces CD8\u03b1 but not CD8\u03b2 expression in DN \u03b3\u03b4 thymic precursors of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs through the upregulation of RUNX family transcription factor 3 (Runx3) and downregulation of Th-inducing POZ-Kruppel factor (Th-Pok), two transcriptional factors important for CD8+T cell commitment in the thymus. Moreover, TGF-\u03b2 directly regulates the maintenance of CD8\u03b1 expression, proliferation, and apoptosis of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs or indirectly influences these \u03b3\u03b4 IELs by modulating the function of IECs. Finally, we found that mice with TGF-\u03b2 signaling deficiency in \u03b3\u03b4T cells were more vulnerable to bacterial attacks and had a worse response to DSS-induced IBD.We here show that TGF-\u03b2 controls the development of TCR\u03b3\u03b418. The number of \u03b3\u03b4 IELs gradually increases from the day of mouse birth and remains stable in 1 month19. We generated mixed BM chimeric mice to investigate the role of TGF-\u03b2 in the development of \u03b3\u03b4 IELs. BM cells from CD45.1 wild-type (WT) mice (CD45.1) were mixed with BM cells from CD45.2 Tgfbr1f/fEsr1-cre mice that had been pretreated with tamoxifen (R1 KO) or oil (R1 WT) for 5 days in a ratio 1:6 (CD45.2:CD45.1). The mixed BM cells were then transferred into irradiated recombination-activation gene 1-deficient (Rag1\u2212/\u2212) mice. The Rag1\u2212/\u2212 mice were sacrificed to examine IELs populations 4\u20135 weeks later cells and reside in the intestine after development in the thymusced Fig. , while tsed Fig. . The fresed Fig. and totased Fig. were alssed Fig. and lympsed Fig. showed nTgfbr2f/fEsr1-cre mice (pretreated with tamoxifen (R2 KO) or oil (R2 WT) for 5 days) and obtained similar results. Specifically, the frequency and total cell number of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs from R2 KO BM-transferred mice were decreased compared to other control mice or WT control BM cells and found that TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs were significantly reduced in Smad2/3dko mice 20. We next examined subsets of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs from Smad2/3dko BM chimeric mice and found that their CD8\u03b1\u03b1+ V\u03b31 \u03b3\u03b4T cell population were all diminished in R1 KO BM-derived thymocytes had no difference between R1 WT and R1 KO mice and \u03b1E\u03b27 integrin , and \u03b3\u03b4T cells in the thymus express a high level of CCR9 that is downregulated when they arrive in the intestine accompanied by CD103 upregulation in mice22. We examined CCR9 on thymic DN4 \u03b3\u03b4T cells from CD45.1\u2009+\u2009R1 WT and CD45.1\u2009+\u2009R1 KO mixed BM chimeric mice. Both the frequency of CCR9+ cells and MFI of CCR9 expression in DN4 thymic \u03b3\u03b4T cell precursors analysis of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs isolated from the mixed BM chimeric mice 4\u20135 weeks post-BM transplantation as shown in Fig. https://www.ncbi.nlm.nih.gov/sra/?term=PRJNA739380). Consistent with our flow cytometry analysis, the gene expression of CD103 (Itgae) was substantially downregulated in TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs from R1 KO BM chimeric mice, while the gene of CCR9 (Ccr9) was comparable is another factor that potentially impacts the migration of \u03b3\u03b4 IELs, and RNA-seq revealed no difference of Itga4 in TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs was downregulated in thymic \u03b3\u03b4T cells by TGF-\u03b21 , Aatk (apoptosis-associated tyrosine kinase), and Bcl2l14 were all upregulated in R1 KO TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs according to our RNA-seq data or tamoxifen (R1 KO)-treated Tgfbr1f/fEsr1-cre mice for 5 days and examined the mRNA expression of Butyrophilin-like (Btnl1), IL-15 and Myd88 in IECs. These genes are expressed on IECs and suggested to be important for TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs generation28. In R1 KO IECs, the expression of Btnl1 or Smad3\u2212/\u2212 mice with IECs from WT or Smad3\u2212/\u2212 mice with a ratio 1:10 and RegIII\u03b2 , such as Il17a, Il23a, and Ifng31 were all upregulated in TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs in TGF-\u03b2 receptor I-deficient mice approach. The intestinal structure of Smad3\u2212/\u2212 mice exhibited more damage with heavier bacteria located within or under the epithelial layer than WT mice that contained fewer TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs was diminished in the absence of TGF-\u03b2 signaling, but CD44\u2212CD25+TCR\u03b3\u03b4+ thymocytes (DN3-\u03b3\u03b4T) were not much altered. This suggests that TGF-\u03b2 might start working in the stage when \u03b3\u03b4T cells are developing from DN3 to DN4. Intriguingly, we found more CD44+CD25\u2212TCR\u03b3\u03b4+ thymocytes, which exhibit DN1-like phenotype, in TGF-\u03b2 receptor I-deficient mice, suggesting that TGF-\u03b2 signaling controls this subset of CD44+CD25\u2212TCR\u03b3\u03b4+ thymocytes37. However, the mechanism underlying this abnormal increase in this unique type of CD44+CD25\u2212TCR\u03b3\u03b4+ thymocytes remains unknown. As TCR\u03b3\u03b4+ thymocytes are presented as either DN3 or DN4 by surface staining38, these CD44+CD25\u2212TCR\u03b3\u03b4+ cells are likely recirculated peripheral \u03b3\u03b4T cells, which are activated in the absence of TGF-\u03b2 or are converted from DN4 CD44\u2212CD25\u2212 \u03b3\u03b4T cells by regaining expression of CD44 due to the Tgfbr1 deficiency. Nevertheless, the decrease in CD44\u2212CD25\u2212TCR\u03b3\u03b4+ DN4 thymocyte should contribute to the deficiency of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs in the absence of TGF-\u03b2 signaling.Consistent with the crucial role of TGF-\u03b2 in the thymic environment+CD8\u03b1\u03b1+ IELs was not affected by TGF-\u03b2 because their expression of the essential chemokine CCR9 and integrin \u03b14\u03b27 for homing to the gut intraepithelial layer was comparable between WT and TGF-\u03b2-deficient mice. However, the ability of \u03b3\u03b4 IELs to reside in the gut could be degraded due to CD103 low expression on TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IEL in TGF-\u03b2 signaling-deficient mice. As \u03b3\u03b4 IELs normally express CD103 rather than CCR9 in the gut, CD103 is more important for \u03b3\u03b4T cells to settle down in the gut. This notion is further supported by the evidence that TGF-\u03b2 induces more expression of CD103 in normal thymic \u03b3\u03b4T cells and gut T cells23. Collectively, TGF-\u03b2 promotes the development of the thymic precursor CD25\u2212CD44\u2212TCR\u03b3\u03b4+ cells of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs and increases the expression of their gut-resident integrin CD103, which together provide a crucial step to fulfill a sufficient number of \u03b3\u03b4 IELs.The migration ability of thymic \u03b3\u03b4 precursors of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs in the gut. Therefore, we have revealed that TGF-\u03b2 induces expression of CD8\u03b1 in thymic \u03b3\u03b4T cells and maintains CD8\u03b1 expression in mature TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs. For thymic \u03b3\u03b4T cells, TGF-\u03b2 induces CD8\u03b1 by regulating the balance of Runx3 and Th-Pok expression, i.e., via upregulation of Runx3 and downregulation of Th-Pok, although the detailed molecular mechanism by which TGF-\u03b2 regulates these two transcriptional factors remains to be elucidated. Strikingly, we believe that the role of TGF-\u03b2 in thymic \u03b3\u03b4T cells also includes facilitating those cells to maintain the potential to develop into CD8\u03b1\u03b1+ homodimers, instead of CD8\u03b1\u03b2+ heterodimers, based on our data that CD8\u03b2 on thymic \u03b3\u03b4T cells tends to be inhibited by TGF-\u03b2 treatment. In splenic \u03b3\u03b4T cells, however, TGF-\u03b2 is capable of driving both CD8\u03b1 and CD8\u03b2 upregulation, suggesting that splenic \u03b3\u03b4T cells are unlikely the precursors of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs. For \u03b3\u03b4 IELs, TGF-\u03b2 upregulates or maintains CD8\u03b1 expression on TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs but does not induce it in TCR\u03b3\u03b4+CD8\u03b1\u2212\u03b2\u2212 IELs. This indicates that TGF-\u03b2 promotes the development of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs not through the conversion of TCR\u03b3\u03b4+CD8\u03b1\u2212\u03b2\u2212 IELs into TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs.However, the aforementioned function of TGF-\u03b2 in the thymic \u03b3\u03b4T precursors clearly cannot explain the unique phenotype of TCR\u03b3\u03b4\u2212CD8\u2212\u03b3\u03b4T cells in the thymus, spleen, and IEL have the distinct potential to be CD8\u03b1+ under stimulation of TGF-\u03b21. In this regard, TGF-\u03b21 increases CD8\u03b1 but decreases CD8\u03b2 in thymic \u03b3\u03b4T cells, upregulates both CD8\u03b1 and CD8\u03b2 in splenic \u03b3\u03b4T cells, and fails to induce CD8\u03b1 and CD8\u03b2 in TCR\u03b3\u03b4+CD8\u03b1\u2212\u03b2\u2212 IELs. Though the underlying mechanisms remain unknown, it could be due to the developing stage and tissue-specific imprint. Thymic \u03b3\u03b4T cells are less developed compared to splenic \u03b3\u03b4T cells or \u03b3\u03b4 IELs and, therefore, have a higher potential to be induced to express CD8\u03b1+. Splenic \u03b3\u03b4T cells, however, tend to express CD8\u03b1\u03b2+ rather than CD8\u03b1\u03b1+ in response to sufficient TGF-\u03b2 stimulation. The biological significance of this feature remains unknown, but it suggests that splenic \u03b3\u03b4T cells are unlikely to be the precursors of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs. TCR\u03b3\u03b4+CD8\u03b1\u2212\u03b2\u2212 IELs should go through a similar developmental process to TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs but cannot express CD8\u03b1\u03b1, suggesting that they are stable in the CD8\u03b1\u2212\u03b2\u2212 phenotype and might be terminally differentiated and, thus, less likely convert to be CD8\u03b1+ when encountering stimuli such as TGF-\u03b2. As expected39, we showed that the decrease in TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs in the absence of TGF-\u03b2 signaling was not due to the suppression of proliferation but can be contributed by the increase in the apoptosis of the knockout TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs.Intriguingly, CD4+ phenotype that is distinct from \u03b3\u03b4T cells in other tissues4. As a crucial cytokine produced by IECs, TGF-\u03b2 also indirectly modulates the maintenance and proliferation of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs by regulating the expression of several genes on IECs, which are known to tightly influence the development of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs40. Btnl is expressed on IECs and determines the maturation and expansion of V\u03b37+ IELs extrathymically regardless of food antigen and microbiomes;19 IL-15 and IL-15R\u03b1 complexes are abundant in IECs and critical for proliferation and survival of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs; MyD88-deficient mice also have less TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs due to IL-15 shortage42. We showed that the expression of Btnl, IL-15, and MyD88 on IECs are all downregulated in TGF-\u03b2 signaling-deficient mice. Reduction of the expression of these genes led Smad3\u2212/\u2212 IECs unable to maintain CD8\u03b1\u03b1+ on TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs in an IECs\u2013IELs coculture system. However, TCR\u03b3\u03b4+CD8\u03b1\u2212\u03b2\u2212 IELs were rarely to be CD8\u03b1+ even co-cultured with WT IECs, further confirming their inability to become TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs. Both TCR\u03b3\u03b4+CD8\u03b1\u03b1+ and TCR\u03b3\u03b4+CD8\u03b1\u2212\u03b2\u2212 IELs showed greater proliferation when co-cultured with Smad3\u2212/\u2212 IECs, suggesting that factors promoting \u03b3\u03b4 IELs proliferation were released more from Smad3\u2212/\u2212 IECs, although this remains to be fully understood. \u03b3\u03b4 IELs developed in an intestinal environment lacking Smad3 show a lower frequency of CD8\u03b1\u03b1+, V\u03b37, and V\u03b34 populations, indicating that the interaction between IECs and IELs is also important for the development and/or function of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs.In addition, IECs provide an organ-specific environment for \u03b3\u03b4 IELs to obtain a special CD8\u03b1\u03b120. As shown previously44, TGF-\u03b2 signaling in \u03b3\u03b4 IELs is crucial for intestinal homeostasis and the control of DSS-induced IBD. Supporting this conclusion is the observation that depletion of TGF-\u03b2 signaling in \u03b3\u03b4T IELs leads to increased susceptibility to and exacerbation of DSS-induced colitis, as evidenced in both Smad3\u2212/\u2212 mice and \u03b3\u03b4T cell-specific TGF-\u03b2 receptor I-deficient mice, due to enhanced bacterial invasion and damage to the epithelial barrier resulting from the reduction of \u03b3\u03b4 IELs in these knockout mice. Furthermore, TGF-\u03b2-deficient TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs exhibit upregulation of pro-inflammation cytokines IFN-\u03b3, IL-6, IL-21, IL-23\u03b1, and IL-12\u03b1, as well as downregulation of antimicrobial proteins such as RegIII\u03b3 and RegIII\u03b231. In addition to its importance in maintaining the integrity of the murine intestinal barrier, TGF-\u03b2 has been shown to be effective in enhancing the cytotoxicity of expanded human V\u03b42 cells in combination with IL-15, likely through the upregulation of CD103 and IL-9 expression in the presence of TGF-\u03b246.Due to their special localization, rapid activation, and broad antigen recognition spectrum, \u03b3\u03b4 IELs serve as the first line of the intestinal immune system to detect bacteria invasion and maintain the integrity of the epithelial barrier+CD8\u03b1\u03b1+ IELs are CD8\u03b1\u03b1+V\u03b37+ and CD8\u03b1\u03b1+V\u03b31+, suggesting that TGF-\u03b2 has the potential to alter the TCR repertoire and modulate the differentiation of \u03b3\u03b4 IELs. This hypothesis is further supported by the weaker ability of \u03b3\u03b4 IELs lacking TGF-\u03b2 signaling to maintain the integrity and homeostasis of the intestinal barrier, indicating that the remaining \u03b3\u03b4 IELs without TGF-\u03b2 exhibit distinct characteristics that may be caused by differences in TCR repertoire. However, we have yet to determine the detailed TCR repertoire specificity of \u03b3\u03b4 IELs lacking TGF-\u03b2 signaling compared to cells from WT mice. We plan to investigate this issue in future studies.According to our results, the main diminished populations of TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs, which play a crucial role in maintaining immune homeostasis and resisting DSS-induced inflammation in the gut.In summary, we have elucidated a previously unrecognized mechanism in which TGF-\u03b2 is a key factor in the formation and development of TCR\u03b3\u03b4Rag1\u2212/\u2212, CD45.1, TCR\u03b4 ER Cre mice used in this study were purchased from The Jackson Laboratory. Tgfbr1f/fEsr1-cre, Tgfbr2f/fEsr1-cre, Smad2f/fER Cre, Smad3\u2212/\u2212, Smad2/Smad3 double KO mice , and Tgfbr1f/fTCR\u03b4ERCre (Tgfbr1f/fcrossed with TCR\u03b4 ER Cre) mice were housed and bred in the animal facility in National Institute of Dental and Craniofacial Research. All mice were housed in specific pathogen-free conditions with adequate food and water supply, with 12\u2009h light/dark cycle, 50% humidity, and room temperature between 25 to 27\u2009\u00b0C, and the maximum number of mice in each cage is 5. All procedure of animal studies was performed under the National Institutes of Health guidelines for the use and care of live animals and were approved by the Animal Care & Use Committee (ACUC) of the National Institute of Dental and Craniofacial Research (NIDCR).6- to 8-week-old C57BL/6, TM Foxp3/Transcription Factor Staining Buffer Set (Cat# 00-5523-00) were from eBioscience. Mouse TCR \u03b3/\u03b4 T cell Isolation Kit (130-092-125) was from Miltenyi Biotec.Anti-mouse CD45.1 (A20), anti-mouse CD45.2 (104), anti-mouse CD45 (30-F11), anti-mouse TCR \u03b3/\u03b4 (GL3), anti-mouse TCR \u03b2 (H57-597), anti-mouse CD8\u03b1 (53-6.7), anti-mouse CD8\u03b2 (YTS156.7.7), anti-mouse V\u03b31.1 (2.22), anti-mouse V\u03b31.2 (4B2.9), anti-mouse V\u03b37 (F2.67), anti-mouse Ki67 (16\u2009A8), anti-mouse CD25 (PC61), anti-mouse CD44 (IM7), anti-mouse CD45RB (C363-16A), anti-mouse CD103 (2E7), anti-mouse CCR9 (9B1), anti-mouse IFN-\u03b3 (XMG1.2), anti-mouse TNF-\u03b1 (MP6-XT22), anti-mouse IL-17A (TC11-18H10.1), anti-mouse CD4 (GK1.5), and anti-mouse CD326 (G8.8) were purchased from Biolegend. Purified anti-mouse CD3 (145-2C11) was purchased from Bio X Cell. Recombinant mouse IL-2 (402-ML) and human TGF-\u03b21 (240-B) were purchased from R&D Systems. SB431542 was obtained from Selleckchem (Cat# S1067). Cell Proliferation Dye eFluor\u2122 450 (Cat# 65-0842-90), eBioscience Cells were collected and adjusted into appropriate density for antibody staining. For cell surface marker staining, cells were incubated with antibodies for 20\u2009min at 4\u2009\u00b0C in the dark. Intracellular staining for cytokines detection, cells were treated with PMA (10\u2009ng/mL), ionomycin (250\u2009ng/mL), and Golgi-Plug (diluted by 1:1000) for 4\u2009h at 37\u2009\u00b0C incubator. After being stained with surface markers, cells were fixed with the fixation/permeabilization buffer solution according to the manufacturer\u2019s instructions. Apoptosis of cells was investigated by staining Annexin V and 7-AAD solution diluted with Annexin binding buffer for 30\u2009min at room temperature. Intranuclear staining were performed by using fixation/permeabilization buffer solution according to the manufacturer\u2019s instructions.15. Small intestines were removed from Peyer patches, opened, and washed with 1\u00d7 PBS several times to clean gut content, then cut into four to six pieces and incubated in IEL buffer for 20\u2009min in 37\u2009\u00b0C incubator with shaking. Suspensions were filtered by 70\u2009\u03bcm and 40\u2009\u03bcm strainer, centrifuged on a 44 and 70% percoll density gradient at 1800 rpm for 20\u2009min with brake 0. IELs can be obtained on the layer between 44% and 70% percoll.IELs from mouse small intestines were obtained as described previously19. Mouse small intestines were opened and washed with cold PBS to remove bacteria and gut content. Clean intestines were cut into small pieces and incubated in DMEM supplemented with 5% FBS, 5\u2009mM EDTA and 0.145\u2009mg/mL dithiothreitol for 20\u2009min on a turning wheel. Tissue pieces were transferred into a new tube with 5\u201310\u2009mL medium, with a vortex of 15\u2009s three times to get more epithelial cells; all the media were collected into a container with suspensions, filtered by 70 and 40\u2009\u03bcm strainer, centrifuged 1000 rpm for 10\u2009min at 4\u2009\u00b0C; cells were suspended by lysis buffer for 3\u2009min and washed once by PBS. Stained by flow antibodies for FACS sorting, CD45\u2212CD326+ cells from above were gated and collected for experiments.IECs were isolated in a way as described in the previous publication+Tgfbr1f/fEsr1-cre or Tgfbr2f/fEsr1-cre mice (treated with tamoxifen for 5 days) and CD45.1+ C57BL/6 mice. CD45.2+ and CD45.1+ BM were mixed with a ratio of 1:6, then injected into irradiated (450 rads) Rag1\u2212/\u2212 mice intravenously. Four to five weeks later, mice were sacrificed and cell populations were isolated for staining. Age-matched littermates of Smad3+/+ or Smad3\u2212/\u2212 recipients were irradiated (450 rads) 6\u2009h before BM cell transfer. BM cells were isolated from C56BL/6\u2009J mice and injected intravenously into Smad3+/+ or Smad3\u2212/\u2212 recipients. One month after transfer, IELs were isolated and analyzed by flow cytometry.BM cells were isolated from CD45.2+CD45+TCR\u03b3\u03b4+ cells, and cultured in 96 round plate in the RPMI-1640 complete medium, supplemented with anti-CD3 , IL-2 (10\u2009ng/mL), with or without TGF-\u03b21 , and SB431542 (5\u2009uM). TCR\u03b3\u03b4+CD8\u03b1\u03b1+ IELs and TCR\u03b3\u03b4+CD8\u03b1\u2212\u03b2\u2212 IELs were sorted from IELs by FACSAria Sorter with gating live+CD45+TCR\u03b3\u03b4+CD8\u03b1+\u03b2\u2212 or live+CD45+TCR\u03b3\u03b4+CD8\u03b1\u2212\u03b2\u2212 specifically and cultured in RPMI-1640 complete medium supplemented with anti-CD3, IL-2 (100\u2009U/mL), IL-3 (10\u2009ng/mL), IL-4 (10\u2009ng/mL), and IL-15 (10\u2009ng/mL) as previously described in ref. 19, treated with or without TGF-\u03b21 , or SB431542 (5\u2009uM).Pure mouse \u03b3\u03b4T cells were sorted from mouse splenocytes or thymocytes by FACSAria Sorter with staining live+CD8\u03b1\u03b1+ IELs or TCR\u03b3\u03b4+CD8\u03b1\u2212\u03b2\u2212 IELs were sorted from IELs by FACSAria Sorter with gating live+CD45+TCR\u03b3\u03b4+CD8\u03b1+\u03b2\u2212 or live+CD45+TCR\u03b3\u03b4+CD8\u03b1\u2212\u03b2\u2212, and were mixed with pure intestinal epithelial cells (sorted by gating live+CD45+CD326+ from IECs) as a ratio 1:10 (IELs:IECs), which is the ratio of IELs to IECs in mice gut according to the published studies1. Cells were cultured in 96 round plates in RPMI-1640 medium, which was supplemented with anti-CD3, IL-2, IL-3, and IL-4 for 3 days before being prepared for flow cytometry.TCR\u03b3\u03b4Tgfbr1f/fTCR\u03b4 ER Cre mice at 7\u201312 days of age were cut and incubated in lysis buffer at 50\u2009\u00b0C overnight, then diluted with dH2O for PCR cocktail preparation. The primers used for Tgfbr1f/f mouse strain are F: TTCTGCTAATCCTGCAGTAAAC; R: ACCCTCTCACTCTTCCTGAGT. And the primers for the TCR\u03b4 ER Cre mouse strain are WT R: GCTTCCAAAACACTTGCACA; Common F: GGAGAGTTTTCCTAGCAGCA; Mutant R: ACACCGGCCTTATTCCAAG. PCR products were separated on 2% agarose gel with EtBr staining to distinguish bands of genes.Tails of Hprt (Mm00446968_m1), CD8\u03b1 (Mm01182197_g1), CD8\u03b21 (Mm00438116m1), Runx3 (Mm00490666), Zbtb7b (Mm00784709_s1), Btnl1 (Mm01281669_m1), Il-15 (Mm00434210), MyD88 (Mm00440338), CCL25 (Mm00436443), or Cdh1 (Mm01247357).Total RNA of isolated IECs or 18-h cultured \u03b3\u03b4T cells were extracted by using RNeasy Mini Kit (QIAGEN) according to the instruction of the manufacturer and reversed transcribed by High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). Quantitative real-time PCR was performed using TaqMan assays with primers of Smad3\u2212/\u2212 or age- and gender-matched Smad3+/+ mice were given 3% DSS drinking water continuously for 7 days; body weight and disease activity index were monitored during treatment. Tgfbr1f/fTCR\u03b4 ER Cre and Tgfbr1+/+TCR\u03b4 ER Cre littermates were treated with tamoxifen (10\u2009mg/mL) five times to deplete T\u03b2R1 specifically on \u03b3\u03b4T cells before 3% DSS water treatment. Body weight and disease activity index were monitored during treatment.47. Small intestinal tissues were taken from mice, fixed, and embedded into paraffin before being cut into 5-\u03bcm sections; then they were deparaffinized in two changes of xylene, rehydrated in 95% and 90% ethanol for 10\u2009min respectively; 16\u2009S rRNA bacteria probe: (AminoC6\u2009+\u2009Alexa488) GCTGCCTCCCGTAGGAGT (Eurofins MWG Operon) was diluted to the concentrations of 100\u2009nM to 1\u2009\u03bcM by hybridization buffer and incubated at 50\u2009\u00b0C for 3\u2009h. DAPI was stained for microscope detection.FISH experiments for bacteria invasion in the intestinal barrier were conducted based on the procedures described in the previous publicationt-test was used for the comparison of two independent experimental groups; the comparison of more than two groups was conducted by one-way ANOVA. RNA-seq data were analyzed by DeSeq2 for gene expression and normalization. P value threshold for the statistical difference was 0.05.Statistical analysis was conducted by GraphPad Prism 9 and shown in figure legends. Unpaired two-tailed Student\u2019s Supplementary InformationSupplementary Dataset S1"} +{"text": "The title compound is a phthalimide-protected polyamine with a protonated central nitro\u00adgen atom. The crystal packing features a hydrogen-bond network, a two-coordinated chloride ion, and off-set \u03c0\u2013\u03c0 stacking. 20H18N3O4+\u00b7Cl\u2212\u00b72H2O, is a phthalimide-protected polyamine that was synthesized by a previous method. It was characterized by ESI\u2013MS, 1H NMR, and FT\u2013IR. Crystals were grown from a solution of H2O and 0.1 M HCl. The central nitro\u00adgen atom is protonated and forms hydrogen bonds with the chloride ion and a water mol\u00adecule. The two phthalimide units make a dihedral angle of 22.07\u2005(3)\u00b0. The crystal packing features a hydrogen-bond network, two-coordinated chloride, and off-set \u03c0\u2013\u03c0 stacking.The title compound {systematic name: bis\u00ad[2-eth\u00adyl]aza\u00adnium chloride dihydrate}, C These units point in opposite directions to each other from the perspective of the central nitro\u00adgen atom. The central tetra\u00adhedral nitro\u00adgen atom (NH2) forms hydrogen bonds with a water mol\u00adecule and the chloride ion.The mol\u00adecular structure of the title compound is shown in Fig.\u00a013.b-axis direction \u22efCg (N3/C13\u2013C20) centroid\u2013centroid distance is 4.0143\u2005(7)\u2005\u00c5. A hydrogen-bond network (Table\u00a01A), a water mol\u00adecule (O1W), and a second water mol\u00adecule (O2W). Both water mol\u00adecules also form hydrogen bonds with phthalimide oxygen atoms . The chloride ions form two hydrogen bonds with the protonated amine and a water mol\u00adecule.The crystal structure features off-set \u03c0\u2013\u03c0 stacking between phthalimide groups running along the on Fig.\u00a02. The Cg k Table\u00a01 exists b4.et al., 20162CH2NH2)2 resulted in 1707 hits, while the structure with protonated amines +HN(CH2CH2NH3+)2 resulted in 182 hits. One of these structures is the triprotonated di\u00adethyl\u00adenetri\u00adamine trichloride , 386.1 (M + Na+). 1H NMR \u03b4 ppm 7.75 , 3.75 , 3.0 , 1.60 . FTIR (cm\u22121) = 3326 \u03bd(N\u2014H), 1698 \u03bd(C=O). Crystals suitable for X-ray crystallography were grown by evaporation, with the compound dissolved in a solution of H2O and 0.1 M HCl.Following a previous protocol (Utz 6.Uiso(H) = 1.2Ueq(N). C-bound and water H atoms were positioned geometrically and refined as riding with Uiso(H) = 1.2\u20131.5Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023004565/ex2070sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023004565/ex2070Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023004565/ex2070Isup3.molSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023004565/ex2070Isup4.cmlSupporting information file. DOI: 2264952CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The aim of the present study was to compare the surface morphology alterations, mineral content, and surface roughness of eroded enamel surface versus eroded enamel surface which was preceded by Bioactive Glass 45S5 (BAG45S5) application in both primary and permanent human dentitions.Fifty-two primary teeth and fifty-two permanent teeth were selected. Teeth were randomly divided into 4 groups of twenty-six teeth each. Groups A1 and B1 underwent erosion with 1% citric acid, while groups A2 and B2 were subjected to application of BAG45S5 powder followed by the same erosive conditions as A1 and B1. Measurements were performed by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and surface profilometry. They were used to examine the surface morphology alterations, mineral content, and surface roughness, respectively.p\u2009<\u20090.005) in both human dentitions. Erosion-only groups showed significantly less surface roughness in permanent teeth (p\u2009<\u20090.045). A marked decrease in surface roughness was observed in surfaces receiving BAG45S5, primary teeth (p\u2009<\u20090.001), and permanent teeth (p\u2009<\u20090.001).SEM of enamel which received BAG45S5 showed smoother surface in primary teeth post erosion. EDX analysis showed that enamel exhibited crucial resistance to mineral loss in the group which received BAG45S5 prior to inducing erosion as compared to the induced erosion-only group. This was significant (Bioactive Glass 45S5 proved successful against erosive conditions in both primary and permanent teeth with better performance in the permanent teeth so it can be regarded as a means of prevention.Bioactive Glass 45S5 powder could be used not only to remove stains but also as a prophylactic preventive measure against the multiple episodes of acidic food and beverage consumption in children. Dental erosion is the chronic and progressive, irreversible loss of dental hard tissues caused by a chemical process without any bacterial involvement. It has become a significant clinical challenge in the recent years. This is due to the major lifestyle changes that have led to an increase in the amount and frequency of consumption of acid-containing foods and beverages . Owing tErosion could have dramatic effect on the dental tissues and would result in alterations in surface roughness, changes in surface morphology, and even mineral loss of dental tissues \u20137. TheseBioactive glasses are biocompatible silicate-based materials, containing calcium and phosphate in an amorphous matrix. They undergo a unique biological reaction at the interface stimulating the formation of a chemical bond between living structures and the material itself . These sA study by Dionysopoulos et al. revealed that surface pre-treatment using air abrasion with BAG45S5 may help to prevent enamel surface erosion induced by an acidic drink in bovine teeth . In addiThe current study is a cross-sectional in vitro study, which was conducted in the Pediatric Dentistry and the Oral Biology Departments, Faculty of Dentistry, Pharos University in Alexandria, Alexandria, Egypt. Ethical approval was obtained from the Research Ethics Committee\u2014Pharos University in Alexandria in adherence to the tenets of the Declaration of Helsinki 1964 and its later amendments (# PUA02202207243036) . ParticiSample size was based on 95% confidence level to detect differences in surface roughness and mineral content between eroded enamel surface as opposed to eroded enamel surface preceded by BAG45S5 application. Dionysopoulos et al. reported mean\u2009\u00b1\u2009SD difference in surface roughness of eroded enamel pre-treated with Bioactive Glass\u2009=\u20090.002\u2009\u00b1\u20090.001, whereas it was 0.006\u2009\u00b1\u20090.01 for eroded enamel . The calAge group 6\u201320\u00a0years; normally shed or serially extracted teeth ; sound labial/buccal enamel surface.Labial/buccal enamel surface with cracks; developmental defects; caries; erosion; fillings.Group A1: twenty-six primary teeth undergoing erosion induction only. Subgroups: two teeth assigned for scanning electron microscopy (SEM) (A1-SEM); twelve teeth for energy-dispersive X-ray spectroscopy (EDX) (A1-EDX); twelve teeth for profilometry (A1-Pro)Group A2: twenty-six primary teeth receiving BAG45S5 followed by erosion induction. Subgroups: two teeth assigned for SEM (A2-SEM); twelve teeth for EDX (A2-EDX); twelve teeth for profilometry (A2-Pro)Group B1: twenty-six permanent teeth undergoing erosion induction only. Subgroups: two teeth assigned for SEM (B1-SEM); twelve teeth for EDX (B1-EDX); twelve teeth for profilometry (B1-Pro)Group B2: twenty-six permanent teeth receiving BAG45S5 followed by erosion induction. Subgroups: two teeth assigned for SEM (B2-SEM); twelve teeth for EDX (B2-EDX); twelve teeth for profilometry (B2-Pro) Fig.\u00a0Fifty-two primary teeth were randomly and equally divided into 2 groups A1 and A2. Fifty-two permanent teeth were also randomly and equally divided into the other 2 groups B1, B2. Randomization was completed using a computer-generated list of random numbers using RAND and RANK functions-Excel (MS Office 365). A serial number was given to each tooth indicating its allocation.Collected teeth were kept in 10% formaldehyde solution at 4\u00a0\u00b0C for 7\u00a0days for sterilization . Teeth w(BAG45S5) in the form of Sylc\u00ae powder with particle size ranging from 25 to 120\u00a0\u00b5m for 3\u00a0min, rinsed with distilled water, and stored in artificial saliva (storage medium) for 65\u201380\u00a0min between the 6 cycles for remineralization iii.Surface roughnessThe outcomes of the current study included:Assessment of outcomes was performed at baseline and post erosion for all groups.Scanning electron microscopy (SEM) JSM-IT200 InTouchScope\u2122 Scanning Electron Microscope was used for this purpose. Two teeth from each group were sectioned into buccal and lingual specimens using a double-sided fine grit water-cooled diamond disc yielding 4 specimens. One specimen of each tooth was randomly selected for baseline assessment of ultra-structure surface morphology alterations, while the other one was used for post erosion assessment. All specimens were dehydrated at the time of evaluation by passed through series of 50%, 70%, and 95% ethyl alcohol for 10\u00a0min each and then in absolute alcohol for two changes of 1-h period each. This was followed by drying in vacuum desiccator for 1\u00a0h . The assEnergy-dispersive X-ray spectroscopy (EDX) JSM-IT200 InTouchScope\u2122 Scanning Electron Microscope was used for this purpose. Assigned teeth were well-polished using diamond paste (1-\u03bcm size). Then, they were washed out under running water, dehydrated, and air dried . After dProfilometry using linear scanning stylus; MarSurf PS 10 ISO 16610\u201321, Lt\u2009=\u20091.5\u00a0mm (0.25\u2009\u00d7\u20095), parameters Ra and Rz was used for this purpose. Twelve teeth from each group were assessed for their surface roughness at baseline and then post erosion. Teeth were firstly polished using 1200-grit abrasive sandpaper to smoothen out any irregularities on the enamel surface. Then, they were stored in artificial saliva for 1\u00a0h. For each tooth, three measurements were taken, and an average reading for those three measurements was obtained.. . Qualitative data were described using number and percent. The Kolmogorov\u2013Smirnov test was used to verify the normality of distribution. Quantitative data were described using range mean, standard deviation. Significance of the obtained results was judged at the 5% level. The used tests: Student t-test for normally distributed quantitative variables to compare between two groups and paired t-test for normally distributed quantitative variables to compare between two periods.Data were analysed using IBM SPSS software package version 20.0Group A1-SEM micrograph demonstrated generalized smooth surface architecture of the enamel surface (\u00d7\u2009500) Fig.\u00a0. Group BGroup A2-SEM micrograph was showing microscopic roughness and irregularities in some areas on the enamel surface (\u00d7\u20091500) Fig.\u00a0. Group Bp\u2009<\u20090.001). Carbon (C) (mean\u2009\u00b1\u2009SD) showed an increase post erosion\u2009=\u200951.48\u2009\u00b1\u20092.38\u00a0than baseline\u2009=\u200912.15\u2009\u00b1\u20092.65 which was statistically significant (p\u2009<\u20090.001). Oxygen (O2) (mean\u2009\u00b1\u2009SD) showed a decrease post erosion\u2009=\u200943.47\u2009\u00b1\u20091.54\u00a0than baseline\u2009=\u200948.60\u2009\u00b1\u20091.75\u00a0which was statistically significant (p\u2009<\u20090.001). Fluoride (F) (mean\u2009\u00b1\u2009SD) showed a negligible amount at baseline and post erosion with no significant difference (p\u2009=\u20090.275).The (mean\u2009\u00b1\u2009SD) was at baseline for calcium (Ca) and phosphorus (P)\u2009=\u200924.29\u2009\u00b1\u20091.24 and 14.30\u2009\u00b1\u20090.81\u00a0respectively which decreased post erosion to\u2009=\u20093.20\u2009\u00b1\u20091.99\u00a0and 1.43\u2009\u00b1\u20090.88\u00a0for Ca and P, respectively. This was statistically significant (p\u2009<\u20090.001). C (mean\u2009\u00b1\u2009SD) showed an increase post erosion\u2009=\u200910.85\u2009\u00b1\u20091.26\u00a0than baseline\u2009=\u20099.59\u2009\u00b1\u20092.50\u00a0which was statistically non-significant (p\u2009=\u20090.182). O2 (mean\u2009\u00b1\u2009SD) showed an increase post erosion\u2009=\u200953.01\u2009\u00b1\u20091.68\u00a0than baseline\u2009=\u200948.96\u2009\u00b1\u20092.97\u00a0which was statistically significant (p\u2009<\u20090.001). Fluoride (F) (mean\u2009\u00b1\u2009SD) showed a negligible amount at baseline and post erosion with no significant difference (p\u2009=\u20090.156) (Table The (mean\u2009\u00b1\u2009SD) was at baseline for Ca and P\u2009=\u200926.09\u2009\u00b1\u20091.57\u00a0and 14.82\u2009\u00b1\u20090.77\u00a0respectively which decreased post erosion to\u2009=\u200922.45\u2009\u00b1\u20091.27 and 12.95\u2009\u00b1\u20090.57\u00a0for Ca and P, respectively. This was statistically significant (p\u2009=\u20090.335). Also, for P, (mean\u2009\u00b1\u2009SD) was comparable between baseline and post erosion\u2009=\u200916.36\u2009\u00b1\u20090.56\u00a0and 16.15\u2009\u00b1\u20090.72,\u00a0respectively. This was non-significant (p\u2009=\u20090.520). C (mean\u2009\u00b1\u2009SD) showed a decrease post erosion\u2009=\u200910.27\u2009\u00b1\u20093.56\u00a0than baseline\u2009=\u200913.18\u2009\u00b1\u20091.92 which was non-significant (p\u2009=\u20090.085). O2 (mean\u2009\u00b1\u2009SD) showed an increase post erosion\u2009=\u200935.23\u2009\u00b1\u20094.30\u00a0than baseline\u2009=\u200932.69\u2009\u00b1\u20093.17\u00a0which was non-significant (p\u2009=\u20090.067). F (mean\u2009\u00b1\u2009SD) showed an increase post erosion than baseline\u2009=\u20090.61\u2009\u00b1\u20090.26\u00a0and\u2009=\u20090.37\u2009\u00b1\u20090.22,\u00a0respectively. This was statistically significant (p\u2009=\u20090.004).The (mean\u2009\u00b1\u2009SD) was comparable for Ca between baseline and post erosion\u2009=\u200937.32\u2009\u00b1\u20092.65\u00a0and 37.92\u2009\u00b1\u20094.48,\u00a0respectively. This was non-significant (p\u2009=\u20090.255). Also, for P, (mean\u2009\u00b1\u2009SD) was comparable between baseline and post erosion\u2009=\u200916.78\u2009\u00b1\u20090.61\u00a0and 17.29\u2009\u00b1\u20090.70,\u00a0respectively. This was non-significant (p\u2009=\u20090.064). C (mean\u2009\u00b1\u2009SD) showed a decrease post erosion\u2009=\u20095.83\u2009\u00b1\u20091.68\u00a0than baseline\u2009=\u200910.55\u2009\u00b1\u20091.63\u00a0which was statistically significant (p\u2009<\u20090.001). O2 (mean\u2009\u00b1\u2009SD) showed an increase post erosion\u2009=\u200936.59\u2009\u00b1\u20095.24\u00a0than baseline\u2009=\u200933.93\u2009\u00b1\u20092.94\u00a0which was statistically significant (p\u2009=\u20090.049). F (mean\u2009\u00b1\u2009SD) showed a negligible amount at baseline and post erosion with no significant difference (p\u2009=\u20090.352) (Table The (mean\u2009\u00b1\u2009SD) was comparable for Ca between baseline and post erosion\u2009=\u200938.19\u2009\u00b1\u20092.99\u00a0and 39.49\u2009\u00b1\u20095.26,\u00a0respectively. This was non-significant (p\u2009<\u20090.001). The (mean\u2009\u00b1\u2009SD) of P content change decreased in A1-EDX than that of B1-EDX\u2009=\u2009\u2009\u2212\u200912.87\u2009\u00b1\u20090.47\u00a0and\u2009\u2212\u20091.87\u2009\u00b1\u20090.53,\u00a0respectively. This change was statistically significant (p\u2009<\u20090.001). The (mean\u2009\u00b1\u2009SD) of O2 content change decreased in A1-EDX than that of B1-EDX\u2009=\u2009\u2009\u2212\u20095.13\u2009\u00b1\u20092.01\u00a0and 4.06\u2009\u00b1\u20092.94,\u00a0respectively. This change was statistically significant (p\u2009<\u20090.001). The (mean\u2009\u00b1\u2009SD) of F content change decreased in A1-EDX than that of B1-EDX\u2009=\u2009\u2009\u2212\u20090.11\u2009\u00b1\u20090.34\u00a0and 0.20\u2009\u00b1\u20090.45,\u00a0respectively. This change was non-significant (p\u2009=\u20090.115). The (mean\u2009\u00b1\u2009SD) of C content change increased in A1-EDX than that of B1-EDX\u2009=\u200939.33\u2009\u00b1\u20092.17\u00a0and 1.25\u2009\u00b1\u20093.05,\u00a0respectively. This change was statistically significant (p\u2009<\u20090.001).Post erosion, the (mean\u2009\u00b1\u2009SD) of Ca content change decreased in A1-EDX than that of B1-EDX\u2009=\u2009\u2009\u2212\u200921.09\u2009\u00b1\u20091.15\u00a0and\u2009\u2212\u20093.64\u2009\u00b1\u20090.98, respectively. This change was statistically significant (p\u2009=\u20090.476). The (mean\u2009\u00b1\u2009SD) of P content change decreased in A1-EDX than that of B1-EDX\u2009=\u2009\u2009\u2212\u20090.22\u2009\u00b1\u20091.14\u00a0and 0.51\u2009\u00b1\u20090.86,\u00a0respectively. This change was non-significant (p\u2009=\u20090.159). The (mean\u2009\u00b1\u2009SD) of O2 content change showed comparable change in A1-EDX and B1-EDX\u2009=\u20092.55\u2009\u00b1\u20094.35\u00a0and 2.66\u2009\u00b1\u20094.18,\u00a0respectively. This change was non-significant (p\u2009=\u20090.947). The (mean\u2009\u00b1\u2009SD) of F content showed comparable change in A1-EDX and B1-EDX\u2009=\u20090.24\u2009\u00b1\u20090.23\u00a0and 0.16\u2009\u00b1\u20090.59,\u00a0respectively. This change was non-significant (p\u2009=\u20090.716). The (mean\u2009\u00b1\u2009SD) of C content change increased in A1-EDX than that of B1-EDX\u2009=\u2009\u2009\u2212\u20092.90\u2009\u00b1\u20095.31\u00a0and\u2009\u2212\u20094.72\u2009\u00b1\u20092.04,\u00a0respectively. This change was non-significant (p\u2009=\u20090.340) of Ca content change decreased in A2-EDX than that of B2-EDX\u2009=\u20090.60\u2009\u00b1\u20092.06\u00a0and 1.31\u2009\u00b1\u20093.76,\u00a0respectively. This change was non-significant p\u2009=\u20090.76. The (P\u2009=\u200916.15\u2009\u00b1\u20090.72 and 1.43\u2009\u00b1\u20090.88 respectively (p\u2009<\u20090.001). The mineral content change of C and O2 significantly decreased in A2-EDX than A1-EDX as C\u2009=\u200910.27\u2009\u00b1\u20093.56 and 51.48\u2009\u00b1\u20092.38 respectively (p\u2009<\u20090.001) and O2\u2009=\u200935.23\u2009\u00b1\u20094.30 and 43.47\u2009\u00b1\u20091.54 respectively (p\u2009<\u20090.001). F content change was comparable between groups A2-EDX and A1-EDX\u2009=\u20090.61\u2009\u00b1\u20090.26 and 0.54\u2009\u00b1\u20090.18 respectively which was non-significant (p\u2009=\u20090.454).The mineral content change of Ca and P significantly increased in A2-EDX than A1-EDX as Ca\u2009=\u200937.92\u2009\u00b1\u20094.48 and 3.20\u2009\u00b1\u20091.99 respectively (p\u2009<\u20090.001) and p\u2009<\u20090.001), and P\u2009=\u200917.29\u2009\u00b1\u20090.70 and 12.95\u2009\u00b1\u20090.57 respectively (p\u2009<\u20090.001).The mineral content change of Ca and P significantly increased in B2-EDX than B1-EDX as Ca\u2009=\u200939.49\u2009\u00b1\u20095.26 and 22.45\u2009\u00b1\u20091.27 respectively (2 significantly decreased in B2-EDX than B1-EDX as C\u2009=\u20095.83\u2009\u00b1\u20091.68 and 10.85\u2009\u00b1\u20091.26 respectively (p\u2009<\u20090.001) and O2\u2009=\u200936.59\u2009\u00b1\u20095.24 and 53.01\u2009\u00b1\u20091.68 respectively (p\u2009<\u20090.001). F content change was comparable between groups B2-EDX and B1-EDX\u2009=\u20090.67\u2009\u00b1\u20090.45 and 0.79\u2009\u00b1\u20090.48 respectively which was non-significant (p\u2009=\u20090.537) . (Mean\u2009\u00b1\u2009SD) of B1-Pro readings at baseline was\u2009=\u20090.56\u2009\u00b1\u20090.16, while for post erosion, it was decreased to\u2009=\u20090.28\u2009\u00b1\u20090.06, and this was statistically significant (p0\u2009<\u20090.001). (Mean\u2009\u00b1\u2009SD) of A2-Pro readings at baseline was\u2009=\u20090.58\u2009\u00b1\u20090.07, while for post erosion, it was decreased to\u2009=\u20090.27\u2009\u00b1\u20090.09, and this was statistically significant (p0\u2009<\u20090.001). (Mean\u2009\u00b1\u2009SD) of B2-Pro readings at baseline was\u2009=\u20090.61\u2009\u00b1\u20090.12, while for post erosion, it was decreased to\u2009=\u20090.28\u2009\u00b1\u20090.10, and this was statistically significant (p0\u2009<\u20090.001). The change in surface roughness from baseline to post erosion significantly decreased in B1-Pro than that of A1-Pro\u2009=\u20090.56\u2009\u00b1\u20090.16/0.28\u2009\u00b1\u20090.06 and\u2009=\u20090.58\u2009\u00b1\u20090.16/0.57\u2009\u00b1\u20090.42 respectively (p1\u2009=\u20090.045). The change in surface roughness from baseline to post erosion was similarly decreased in A2-Pro and B2-Pro\u2009=\u20090.58\u2009\u00b1\u20090.07/0.27\u2009\u00b1\u20090.09 and\u2009=\u20090.61\u2009\u00b1\u20090.12/0.28\u2009\u00b1\u20090.10 respectively (p1\u2009=\u20090.563). The change in surface roughness, post erosion, between group A1-Pro\u2009=\u20090.57\u2009\u00b1\u20090.42, and A2-Pro\u2009=\u20090.27\u2009\u00b1\u20090.09 was significantly less in group A2-Pro (p\u2009=\u20090.026). The change in surface roughness, post erosion, between group B1-Pro\u2009=\u20090.28\u2009\u00b1\u20090.06, and B2-Pro\u2009=\u20090.28\u2009\u00b1\u20090.10 was not significant (p\u2009=\u20090.960) (Table (Mean\u2009\u00b1\u2009SD) of A1-Pro readings at baseline was\u2009=\u20090.58\u2009\u00b1\u20090.16, while for post erosion, it was\u2009=\u20090.57\u2009\u00b1\u20090.42 which was not statistically significant p0\u2009=\u20090.55. on both human primary and permanent dentitions.\u00a0To overcome the lack of reliability in the use of bovine teeth instead of human ones, teeth were used from both human dentitions instead of bovine teeth for more favourable clinically applicable results. In a review of literature conducted by Yassen et al. and baseDissolution of the enamel usually occurs when it is exposed to undersaturated solutions at a pH less than the critical pH, which is thought to be 5.5 . TherefoBAG45S5 has been widely studied for its effectiveness on enamel remineralization. It improved the microhardness of the subsurface-eroded enamel surface . A fluorChanges in surface roughness commonly occur after the interaction of acidic solutions with tooth structures. While the effect of erosive solutions on the surface roughness depends on the thickness of the aprismatic layer, the non-treated permanent enamel with its thin aprismatic surface layer demonstrated a notable roughness after exposure to the erosive acid, whereas the non-treated primary enamel with its generally uniform and thicker aprismatic layer presenting a more stable surface texture was not as affected by the erosive agent . On the Moreover, the results of the current study revealed that both human dentitions were showing significant loss of calcium and phosphate ions post the erosive challenge. The dissolution was expected to occur due to the weak constitution of the calcium-deficient carbonated hydroxyapatite crystals present in the enamel layer causing a distorted lattice conformation that is\u202fmore soluble in acidic environments.\u00a0The primary dentition presented greater mineral loss and surface distortion, because of the higher number of carbonate molecules in the hydroxyapatite crystals in the primary enamel as well as the presence of copious organic content specific to primary enamel . This isOn the contrary, enamel surface that was pre-treated by BAG45S5 before the erosive challenge showed decreased rates of carbon ions when compared to the baseline. Moreover, there was no significant change in calcium or phosphate content after erosion also compared to baseline. The effect of BAG45S5 on enamel lessened the effect of erosion on either primary or permanent enamel compared to enamel of erosion-only surfaces. BAG45S5 was suggested to be able to restore early erosive enamel lesions with complete loss of hydroxyapatite crystal content . In the On demineralized enamel surface, BAG45S5 acted in some way differently as suggested by several studies. Abbassy et al. , in theiThroughout the comparison between both dentitions, the permanent dentition which received BAG45S5 showed slightly better response following exposure to induced erosive challenge. Permanent enamel has better consolidation with particles of BAG45S5 due to the intrinsically higher inorganic content as well as morphological differences, indicating that the permanent enamel\u202fallows better integration of Bioactive Glass with the inherent calcium and phosphate ions to form more hydroxycarbonate apatite crystals . Since tIt can be concluded that applying Bioactive Glass 45S5 (BAG45S5) to the enamel surface prior to inducing erosion showed better results than inducing erosion alone in both primary and permanent dentitions. Consequently, BAG45S5 can limit or even prevent dental erosion in both primary and permanent dentitions by protecting the enamel surface from acid dissolution, with better performance on the permanent enamel. This means that BAG45S5 could be considered a promising method of erosion prevention. It could be implemented as a part of the preventive program tailored to children who show high consumption of juice and acidic beverages. Further clinical trials on children are needed to assess the effect on teeth sensitivity of using BAG45S5 prior to the repeated episodes of acidic food and beverage intake.Being dependant on collecting human teeth, difficulty was encountered in collecting the required number of teeth which led to compensating this through teeth sectioning."} +{"text": "Correction to: Acta Neuropathologica (2019) 138:251\u2013273 10.1007/s00401-019-02013-zIn this article author would like to make few changes in the article text. The corrections are given below.1. In Materials and methods section for Immunofluorescence of human sections:In the sentence:goat serum blocking with 0.1\u00a0M KPBS and 0.025% Triton-X) for at least 1\u00a0h with gentle agitation.\u201d\u201cThe sections were then washed (3\u2009\u00d7\u200915\u00a0min) in 0.1\u00a0M KPBS, after which they were incubated in blocking buffer and TNF-\u03b1 (627\u20130.153\u00a0pg/mL). The A\u03b240 detection range was 15,100\u20133.69\u00a0pg/mL, and the A\u03b242 detection range was 2280\u20130.557\u00a0pg/mL. ELISA plates from R&D (DY008 and DY1197-05) were used to measure the levels of gal3 (detection range 1000\u201315.6\u00a0pg/mL) in culture media.\u201d\u201cThe detection ranges of the different cytokines measured were as follows: IL1\u03b2 (1670\u20130.408\u00a0pg/mL), IL4 (1660\u20130.405\u00a0pg/mL), IL12 , IL10 (3410\u20130.833\u00a0pg/mL), IFN-\u03b3 (938\u20130.229\u00a0pg/mL), IL2 (2630\u20130.642\u00a0pg/mL), IL5 (967\u20130.236\u00a0pg/mL), IL6 (5720\u20131.40\u00a0pg/mL), Action: Write \u201cIL8 regarded as human functional homolog to rodent KC\u201d\u00a0between brackets next to KC/GRO."} +{"text": "Analysis of the Hirshfeld surface shows structure-defining inter\u00adactions for bromo\u00admethyl\u00adalcohol 1, resulting in inter\u00admolecular hydrogen bonds between the hydroxyl groups along the a-axis direction.Trimeth\u00adyl({tris\u00ad[(phenyl\u00adsulfan\u00adyl)meth\u00adyl]sil\u00adyl}meth\u00adoxy)silane ( 3), C26H32OS3Si, is a new ligand for transition-metal coordination chemistry derived from 3-bromo-2,2-bis\u00ad(bromo\u00admeth\u00adyl)propan-1-ol (1), C5H9Br3O, through silylation and following exchange of bromine groups with NaSPh. Silylated thio\u00adether ligand 3 crystallizes in the centrosymmetric space group C2/c. Bromo\u00admethyl\u00adalcohol 1 crystallizes in the space group P1, resulting in inter\u00admolecular hydrogen bonds between the hydroxyl groups along the a-axis direction.Trimeth\u00adyl({tris\u00ad[(phenyl\u00adsulfan\u00adyl)meth\u00adyl]sil\u00adyl}meth\u00adoxy)silane ( As a result of the soft property of thio\u00adethers, they coordinate to transition metals . The mol\u00adecular structure of bromo\u00admethyl\u00adalcohol 1 is shown in Fig.\u00a01Bromo\u00admethyl\u00adalcohol et al., 1987The bond lengths to be expected for a C\u2013Br bond are in the range 1.880\u20131.940\u2005\u00c5 % for O1/Si1/C24\u2013C26 and 49.1\u2005(3)% for O1\u2032/Si1\u2032/C24\u2032\u2013C26\u2032. The disorder at the TMSO group also shows a shorter O1\u2014C23 [1.384\u2005(15)\u2005\u00c5] bond length than O1\u2032\u2014C23 [1.479\u2005(14)\u2005\u00c5]. The expected bond length for a C\u2014O\u2014Si bond is between 1.365 and 1.467\u2005\u00c5 bonds are of comparable lengths to each other and correspond to the expected bond lengths for alk\u00adyl\u2013sulfur bonds \u2005\u00c5]. Moreover, the angles are approximately linear at 175\u2005(4)\u00b0 (O3\u2014H3\u22efO4) and 168\u2005(4)\u00b0 (O1\u2014H1\u22efO2). These hydrogen bonds can be assigned the graph-set symbol D11(2). This means that a hydrogen bond between two adjacent hydroxyl groups, O1\u2014H1\u22efO2, is established. The contact between O4\u2014H4\u22efO1 is created via the symmetry operation (i) x\u00a0\u2212\u00a01, y, z.The crystal packing of bromo\u00admethyl\u00adalcohol CrystalExplorer21 , which is due to the spatial arrangement of the bromine substituents in relation to the hydroxyl group.To gain further insight into the supra\u00admolecular packing inter\u00adactions, a Hirshfeld surface analysis was performed prop\u00adoxy]benzene-1,2-dicarbo\u00adnitrile silane]di\u00adbromo\u00admercury(II)] silane-S,S\u2032]mercury(II) -2-[(phenyl\u00adthio)\u00admeth\u00adyl]propan-2-ol yielded no hits in the WebCSD. By replacing the quaternary carbon with a silicon atom, the comparable structural motif of 2I2 rhomboids was dropped into diethyl ether (50\u2005mL) at 273.15\u2005K to 1 (15.39\u2005mmol). The solution was stirred for 1\u2005h at room temperature and then chloro\u00adtri\u00admethyl\u00adsilane (16.93\u2005mmol) was added at 273.15\u2005K. It was stirred again for 1\u2005h at room temperature, then the reaction solution was quenched with water. The aqueous phase was extracted three times with di\u00adchloro\u00admethane and the combined organic layers were dried over magnesium sulfate. The volatiles were removed to give compound 2 crude.Methyl\u00adlithium \u03b4 = 3.33 , 3.18 , 0.03 3) ppm.1H}13C NMR \u03b4 = 61.6 , 44.3 , 34.6 , \u22120.7 ppm.{2 in DMF (10\u2005mL) at 273.15\u2005K and stirred for 10 minutes. The reaction solution was irradiated with microwaves and then quenched with water. The aqueous phase was extracted three times with di\u00adchloro\u00admethane, the combined organic layers were dried over magnesium sulfate and the volatiles were removed. The residue was separated by fractional distillation under reduced pressure. Crystallization from diethyl ether at 243.15\u2005K provided silylated thio\u00adether ligand 3 as clear and colourless blocks.Thio\u00adphenol (5.83\u2005mmol) was then added to sodium hydride (5.83\u2005mmol) in DMF (5\u2005mL) at 273.15\u2005K and stirred for 10 minutes. The NaSPh solution was added to 1H NMR \u03b4 = 7.38\u20137.34 , 7.30\u20137.23 , 7.19\u20137.15 , 3.60 , 3.22 , 0.04 ppm.1H}13C NMR \u03b4 = 137.2 , 129.8 , 128.9 , 126.2 , 64.1 , 46.0 4), 38.7 , \u22120.6 ppm.{6.Uiso(H) = 1.2Ueq(C) for CH2 and CH hydrogen atoms and Uiso(H) = 1.5Ueq(C) for CH3 hydrogen atoms. Hydrogen atoms H1, H2, H3 and H4 for compound 1 were refined independently. The TMSO group in 3 is disordered with occupancies converging to 50.9\u2005(3)% for O1/Si1/C24\u2013C26 and 49.1\u2005(3)% for O1\u2032/Si1\u2032/C24\u2032\u2013C26\u2032.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S205698902300227X/vm2278sup1.cifCrystal structure: contains datablock(s) 1, 3, global. DOI: 10.1107/S205698902300227X/vm22781sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S205698902300227X/vm22783sup3.hklStructure factors: contains datablock(s) 3. DOI: Click here for additional data file.10.1107/S205698902300227X/vm22781sup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698902300227X/vm22783sup5.cmlSupporting information file. DOI: 2247226, 2247225CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title carbazole derivative, the dihedral angle between the carbazole ring system and the pendant phenyl ring is 56.78\u2005(8)\u00b0. 19H15NO, the dihedral angle between the benzene rings of the carbazole moiety is 1.73\u2005(12)\u00b0 and the meth\u00adoxy-substituted phenyl ring deviates from the mean plane of the carbazole grouping (r.m.s. deviation = 0.020\u2005\u00c5) by 56.78\u2005(8)\u00b0. In the crystal, weak C\u2014H\u22ef\u03c0 inter\u00adactions link the mol\u00adecules. The two-dimensional fingerprint plots derived from the Hirshfeld surface indicate that H\u22efH (51.2%) and C\u22efH/H\u22efC (39.9%) contacts dominate the packing.In the title compound, C The crystal structure is consolidated by weak C\u2014H\u22ef\u03c0 inter\u00adactions = 2.71\u2005\u00c5; symmetry code: 1\u00a0\u2212\u00a0x, \u2212y, z; C17\u2014H17\u22efCg(C7\u2013C12) = 2.92\u2005\u00c5; symmetry code: x, 1\u00a0+\u00a0y, z]. The crystal packing viewed along the a-axis direction is shown in Fig.\u00a03The title compound crystallizes in the ortho\u00adrhom\u00adbic space group it Fig.\u00a01. As expens Fig.\u00a02 [C9\u2014H9\u22efCet al., 2017dnorm within the range \u22120.05 to 1.63 a.u. shows a few red spots in the locales of D\u22efA inter\u00adactions, consistent with the presence of weak C\u2014H\u22ef\u03c0 inter\u00adactions , which were uniformly dispersed in 35\u2005ml of di\u00admethyl\u00adformamide (DMF) and the mixture was dissolved homogeneously under an N2 atm. To the mixture, K2CO3 , CuI and 1,10-phenanthroline was added and the mixture was refluxed for 12\u2005h under N2. The progress of the reaction was monitored by TLC. After the completion of the reaction, the reaction was quenched in ice\u2013water and the solid product was dissolved in ethyl acetate and washed with brine solution. The obtained solvent was removed under reduced pressure and the obtained residue was further purified using column chromatography (100\u2013200 mesh silica gel) to afford the title compound as shown in Fig.\u00a06A 100\u2005ml round-bottom flask was charged with 4-iodo\u00adanisole 1.78\u2005g , 9Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2414314623006740/hb4441sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314623006740/hb4441Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623006740/hb4441Isup3.cmlSupporting information file. DOI: 2280833CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structures of two benzo\u00adthio\u00adphene derivatives are described and the inter\u00admolecular contacts in the crystals analysed using Hirshfeld surface analysis and two-dimensional fingerprint plots. 22H16FNO2S2 (I) and C26H20FNO2S2 (II), the benzo\u00adthio\u00adphene rings are essentially planar with maximum deviations of 0.009\u2005(1) and 0.001\u2005(1)\u2005\u00c5 for the carbon and sulfur atom in compounds I and II, respectively. In I, the thio\u00adphene ring system is almost orthogonal to the phenyl ring attached to the sulfonyl group, with a dihedral angle of 77.7\u2005(1)\u00b0. In compound I, the mol\u00adecular structure is stabilized by weak C\u2014H\u22efO intra\u00admolecular inter\u00adactions formed by the sulfone oxygen atoms, which generate two S(5) ring motifs. In the crystal of I, N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into R22(8) rings, which are connected into a C(10) chain via C\u2014H\u22efF hydrogen bonds. Inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions are also observed. In compound II, the mol\u00adecules are linked via C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonding, generating infinite C(11) and C(13) chains running parallel to [010].In the title compounds, C Thio\u00adphene derivatives possess pharmacological and biological activities including anti\u00adbacterial benzene\u00adsulfonamide (C7\u2013C22/N1/S2/O1/O2/F1) while compound II, C26H20FNO2S2 benzene\u00adsulfonamide (C9\u2013C15/N1/S2/O1/O2). In both compounds, the benzo\u00adthio\u00adphene ring systems (S1/C1\u2013C8) are essentially planar with maximum deviations of 0.009\u2005(1) and 0.001\u2005(1)\u2005\u00c5 for atom C8 and S1 in compounds I and II, respectively. The mean planes of the thio\u00adphene ring system in I make dihedral angles of 1.2\u2005(2), 2.3\u2005(2), 77.7\u2005(2)\u00b0 with the C1\u2013C6, C11\u2013C16 and C17\u2013C22 phenyl rings. The mean planes of the thio\u00adphene ring system in II make dihedral angles of 0.3\u2005(2), 33.3\u2005(2), 25.2\u2005(2)\u00b0, respectively, with the C1\u2013C6, C11\u2013C16 and C17\u2013C22 phenyl rings, The benzo\u00adthio\u00adphene ring system in I is almost orthogonal to the C17\u2013C22 ring attached to sulfonyl group with dihedral angle of 77.7\u2005(1)\u00b0 in I. For both compounds, the bond lengths and angles are close to those observed for similar structures \u00b0 in II], with a simultaneous decrease in the N1\u2014S2\u2014C17 angle [106.35\u2005(9) in I and 108.3\u2005(2)\u00b0 in II] from the ideal tetra\u00adhedral value (109.5\u00b0) are attributed to the Thorpe\u2013Ingold effect and N1\u2014C16 = 1.450\u2005(5)\u2005\u00c5 bond lengths in the mol\u00adecule are longer than the mean Nsp2\u2014Csp2 bond-length value of 1.355\u2005(14)\u2005\u00c5 inter\u00adactions, where the Cg1 is the centroid of the C1\u2013C6 ring were performed with CrystalExplorer17 through white to blue . The dnorm surface was mapped over a fixed colour scale of \u22120.434 (red) to 1.449 (blue) for compound I and \u22120.119 (red) to 1.765 (blue) for compound II, where the red spots indicate the inter\u00admolecular contacts involved in the hydrogen bonding. The electrostatic potential was also mapped on the Hirshfeld surface using a STO-3G basis set and the Hartee\u2013Fock level of theory . Areas on the Hirshfeld surface with high curvedness tend to divide the surface into contact patches with each neighbouring mol\u00adecule. The coordination number in the crystal is defined by the curvedness of the Hirshfeld surface . The nearest neighbour coordination environment of a mol\u00adecule is identified from the colour patches on the Hirshfeld surface depending on their closeness to adjacent mol\u00adecules .The Hirshfeld surfaces of compounds I, they reveal that the principal inter\u00admolecular contacts are H\u22efH contacts at 36.9% , H\u22efC/C\u22efH contacts at 26.1% , O\u22efH/H\u22efO at 15.1% , F\u22efH/H\u22efF at 9.2% , C\u22efC at 6.7% , S\u22efC/C\u22efS at 2.2% , S\u22efH/H\u22efS contacts at 0.9% , F\u22efC/C\u22efF at 0.8% , N\u22efC/C\u22efN at 0.7% and N\u22efH/H\u22efN contacts at 0.3% .The fingerprint plots are given in Figs. 89% Fig.\u00a08b, H\u22efC/C1% Fig.\u00a08c, O\u22efH/H1% Fig.\u00a08d, F\u22efH/H2% Fig.\u00a08e, C\u22efC a7% Fig.\u00a08f, S\u22efC/C2% Fig.\u00a08g, S\u22efH/H9% Fig.\u00a08i, F\u22efC/C8% Fig.\u00a08j, N\u22efC/C7% Fig.\u00a08k and N\u22ef3% Fig.\u00a08l.II, they reveal a similar trend, with the principal inter\u00admolecular contacts being H\u22efH/H\u22efH at 41.4% , H\u22efC/C\u22efH contacts at 25.1% , O\u22efH/H\u22efO at 12.1% , F\u22efH/H\u22efF at 8.1% C\u22efC at 4.6% , C\u22ef\u00b7C at 4.7% , S\u22efH/H\u22efS contacts at 4.5% , S\u22efC/C\u22efS contacts at 2.1% , C\u22efO/O\u22efC contacts at 1.0% , F\u22efS/S\u22efF at 0.9% and O\u22efO contacts at 0.3 . In both compounds, the H\u22efH inter\u00admolecular contacts predominate, followed by the C\u22efH/H\u22efC and O\u22efH/H\u22efO contacts.For compound 4% Fig.\u00a09b, H\u22efC/C1% Fig.\u00a09c, O\u22efH/H6% Fig.\u00a09e, C\u22ef\u00b7C 7% Fig.\u00a09f, S\u22efH/H5% Fig.\u00a09g, S\u22efC/C1% Fig.\u00a09h, C\u22efO/O0% Fig.\u00a09i, F\u22efS/S9% Fig.\u00a09j and O\u22ef.3 Fig.\u00a09k. In bo5.I: To a solution of (E)-2-(2-(benzo[b]thio\u00adphen-2-yl)vin\u00adyl)-5-fluoro\u00adbenzenaminium chloride in dry DCM (10\u2005mL), pyridine and PhSO2Cl were added and stirred at room temperature for 12\u2005h. After completion of the reaction (monitored by TLC), it was poured into crushed ice (50\u2005g) containing conc. HCl (5\u2005mL), extracted with DCM (2 \u00d7 20\u2005mL) then washed with water (2 \u00d7 20\u2005mL) and dried (Na2SO4). Removal of solvent in vacuo followed by crystallization from di\u00adethyl\u00adether (4\u2005mL) afforded (E)-N-{2-[2-(benzo[b]thio\u00adphen-2-yl)ethen\u00adyl]-5-fluoro\u00adphen\u00adyl}benzene\u00adsulf\u00adonamide as a white solid.Compound II: To a solution of (E)-N-{2-[2-(benzo[b]thio\u00adphen-2-yl)vin\u00adyl]-5-fluoro\u00adphen\u00adyl}benzene\u00adsulfonamide in CH3CN (10\u2005mL), K2CO3 and 1-bromo\u00adbut-2-yne were added and stirred at room temperature for 12\u2005h. After completion of the reaction (monitored by TLC), it was poured into crushed ice (50\u2005g) containing conc. HCl (5\u2005mL), extracted with ethyl acetate (2 \u00d7 20\u2005mL) then washed with water (2 \u00d7 20\u2005mL) and dried (Na2SO4). Removal of solvent in vacuo followed by crystallization from methanol (4\u2005mL) afforded (E)-N-{2-[2-(benzo[b]thio\u00adphen-2-yl)ethen\u00adyl]-5-fluoro\u00adphen\u00adyl}-N-(but-2-yn-1-yl)benzene\u00adsulfonamide as a white solid.Compound 6.I, the NH H atoms were located in difference-Fourier maps and freely refined. For compound II, they were included in calculated positions and refined as riding: N\u2014H = 0.93\u2005\u00c5 with Uiso(H) = 1.2Ueq(N). All C-bound H atoms were positioned geometrically and constrained to ride on their parent atoms: C\u2013H = 0.93\u20130.97\u2005\u00c5 with Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. In compound I, the thio\u00adphene ring is disordered over two positions with a refined occupancy ratio of 0.756\u2005(4):0.244\u2005(3). The geometries were regularized using soft restraints.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023006096/zn2029sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989023006096/zn2029Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989023006096/zn2029IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989023006096/zn2029Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023006096/zn2029IIsup5.cmlSupporting information file. DOI: 2280606, 2280605CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "It crystallizes isotypically with H-La2[Si2O7]. The four crystallographically distinct CeIII cations form distorted square anti\u00adprisms, capped square anti\u00adprisms, and bicapped square anti\u00adprisms as coordination polyhedra consisting of oxygen atoms. Four crystallographically different silicon atoms recruit the centers of two different isolated [Si2O7]6\u2013 units.The title compound, dicerium(III) oxidodisilicate, Ce H-type Ce2[Si2O7], like H-La2[Si2O7] cover an inter\u00adval from 2.366\u2005(4) to 2.817\u2005(4)\u2005\u00c5 (Table\u00a01plus 3.11\u2005(4)\u20133.34\u2005(4)\u2005\u00c5 to most caps. All oxygen atoms belong to pyroanionic oxidodisilicate anions [Si2O7]6\u2013 = 1.588\u2005(4)\u20131.676\u2005(4)\u2005\u00c5 as reaction vessel at a temperature of 1023\u2005K, taking advantage of the presumed mineralizers Sb2O3 and CeCl3. The transparent, colorless crystals exhibit a platelet-like habit.Single crystals of Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314623005916/wm4192sup1.cifCrystal structure: contains datablock(s) I, 1R. DOI: 10.1107/S2414314623005916/wm4192Isup2.hklStructure factors: contains datablock(s) I. DOI: 2222660CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Araucaria araucana is an ancient conifer, native to the mountain ranges in Chile and Argentina. These trees host a large number of organisms, mainly insects, strongly or even exclusively associated with them. The recent emergence of a novel canker disease on A. araucana has emphasised the importance of fungi associated with these iconic trees and has resulted in the discovery of various new species. In this study, we considered the identity of an unknown calicioid fungus consistently found on resin on the branches of A. araucana. Preliminary phylogenetic analyses placed isolates in the recently described sub-class Cryptocaliciomycetidae, closest to Cryptocalicium blascoi. However, the morphology of the ascomata and its occurrence in a unique niche suggested that the closest relative could be Resinogalea humboldtensis , a fungus with similar sporing structures found on resin of Araucaria humboldtensis in New Caledonia. There are no living cultures or sequence data available for either R. humboldtensis or its supposed closest relative, Bruceomyces castoris, precluding sequence-based comparisons. Morphological comparisons of the sporing structures on A. araucana confirmed that the ascomatal morphology of our unknown calicioid fungus and R. humboldtensis are almost identical and resemble each other more so than B. castoris or Cr. blascoi. A phylogenetic analysis based on the small subunit (SSU), internal transcribed spacer (ITS) and large subunit (LSU) rDNA regions resolved our strains into two clades with Cr. blascoi as its closest relative. Further analyses applying the Genealogical Concordance Phylogenetic Species Recognition (GCPSR) based on ITS, mini chromosome maintenance protein complex (MCM7), RNA polymerase II second largest subunit (RPB2) and translation elongation factor 1-alpha (TEF) gene regions, confirmed that strains represent two new species. Based on our morphological observations and phylogenetic analyses, we introduce two new Resinogalea species, R. araucana and R. tapulicola, and reclassify the genus in the subclass Cryptocaliciomycetidae.The online version contains supplementary material available at 10.1186/s43008-023-00122-9. Araucaria araucana (Araucariaceae), commonly known as Araucaria, pew\u00e9n or the monkey puzzle tree, is an ancient species native to the mountain ranges of Chile and Argentina. The species is one of only two in the genus and family found in South America, with the remainder distributed in the Southeast Asia\u2013Oceania region to accommodate both genera, Resinogalea and Bruceomyces, each containing a single species or 2% malt extract agar in Petri dishes. Plates were incubated for 14 d at 20\u201325\u00a0\u00b0C. Fungal colonies of interest were transferred to new MEA plates in order to produce pure cultures. Isolates were further purified by transferring them to 2% water-agar and allowing these to grow for 2\u20134 d before transferring single hyphal tips to new MEA plates. Direct isolations from ascomata were made by collecting ascospores with a sterile needle, spreading these onto WA and after 2\u20134\u00a0days, transferring germinating single spores to new MEA plates.All relevant isolates used in this study were deposited into the culture collection (CMW) housed at the Forestry and Agricultural Biotechnology Institute . Representative strains for each species were subsequently deposited into the CMW-IA culture collection and to the culture collection (CBS) of the Westerdijk Fungal Biodiversity Institute . Specimens were deposited in the PREM fungarium of the South African National Collections of Fungi housed at the Agricultural Research Council .DNA extractions were performed using the Prepman\u00ae Ultra Sample Preparation Reagent kit following the manufacturer\u2019s protocols. For some strains, DNA was extracted using a phenol\u2013chloroform method. For the latter approach, strains were grown on MEA for 21 d, after which mycelium was collected in 2\u00a0mL Eppendorf tubes and freeze dried overnight. Mycelia was ground by adding metal beads and shaking this in a mixer mill and then following the protocol described by Barnes et al. and modiRPB2) with primers RPB2-5f2 & RPB2-7cR with primers Cer-MCM7F & Cer-MCM7R large intron with primers EF1-782F & EF-2 of each primer (10\u00a0\u00b5M), 0.3 \u00b5L MyTaq\u2122 DNA Polymerase and 17.7 \u00b5L (16.7 \u00b5L for MCM7) sterile deionized water. The thermal cycling conditions included an initial denaturation step of 95 \u02daC for 3\u00a0min, followed by 35 cycles of denaturing at 95 \u02daC for 30\u00a0s, annealing at 56 \u02daC (52 \u02daC for TEF1 and MCM7) for 30\u00a0s and elongation at 72 \u02daC for 45\u00a0s, followed by a final elongation step at 72 \u02daC for 4\u00a0min. Successful amplifications were confirmed by staining PCR amplicons with GelRed\u00ae (2 \u00b5L per 4 \u00b5L of PCR product) and electrophoresing the product on a 1% agarose gel for 12\u00a0min at 110\u00a0V.Six gene regions were amplified through PCR, which included: (i) the internal transcribed spacer 5.8S rDNA (ITS) with primers ITS1 & ITS4 .PCR amplicons were cleaned from excess primers and unincorporated nucleotides either by sodium acetate precipitation approach and edited using Affinity Designer v.1.10.5.1342 (Serif (Europe), Nottingham, UK). Aligned datasets and IQ-TREE outputs were uploaded to the University of Pretoria\u2019s research data repository hosted on Figshare and can be accessed at https://doi.org/10.25403/UPresearchdata.21640298.Each dataset was aligned using the MAFFT v.7 online service selecting the L-INS-i algorithm . Microscopic slides were prepared from 28 d old colonies using 10% lactic acid as mountant. Structures collected directly from samples were first sectioned using a Leica CM1520 cryostat before a microscope specimen was prepared. Characters were observed and photographed using a Zeiss AXIO Zoom.V16 dissection and AXIO Imager.A2 compound microscopes both equipped with a Zeiss AxioCaM 512 colour camera driven by Zen Blue v.3.2 software . For dissection microscope images, extended depth of field analyses were performed using Helicon Focus v.7.5.4 . Up to 50 measurements were made for each character where available, and boxplot graphs were constructed with measurements of each character to visualize data distribution. Photographic plates were prepared in Affinity Designer v.1.10.5.1342 (Serif (Europe) Ltd, Nottingham, UK). Attempts were made at the time of the study to obtain specimens for R. humboldtensis but these were unsuccessful. Morphological comparisons were thus based on the description published in Rikkinen et al. using a micropipette and allowed to dry for 10\u00a0min before plates were sealed with Parafilm\u00ae and incubated upside down. Plates were incubated at temperature gradient of 5\u00a0\u00b0C intervals from 5\u201335\u00a0\u00b0C for 21 d and colony diameters measured every 3\u20134 d. The colony area was measured from images taken on day 21 using ImageJ v.1.52 and Trongol Alto .Isolations made from surface disinfested n\u2009=\u200923), and from single spore isolates from five fruiting bodies (n\u2009=\u20095). A preliminary phylogenetic analysis , 39 gaps (6%)), followed by Penicillium scabrosum , 35 gaps (5%)), P. hirayamae , 28 gaps (4%)), P. copticola\u00a0, 16 gaps (3%)), and P. terrigenum , 27 gaps (4%)). Six strains were selected from each of the two ITS clades and five additional gene regions sequenced for them to continue further analyses.DNA extraction and ITS sequencing was performed for all strains originating from plant tissue , ITS and LSU were concatenated resulting in a matrix containing 62 taxa that was 5770\u00a0bp long. The most suitable nucleotide substitution models according to the Bayesian information criterion (BIC) for SSU, ITS and LSU were TNe\u2009+\u2009I\u2009+\u2009G4, GTR\u2009+\u2009F\u2009+\u2009I\u2009+\u2009G4, and TN\u2009+\u2009F\u2009+\u2009I\u2009+\u2009G4 respectively. ML and BI analyses resolved strains, with high bootstrap support, into two clades in the subclass Cryptocaliciomycetidae with Cr. blascoi as its closest relative in Cr. blascoi and the outgroup Caliciopsis pinea. Substitution models for the aligned ITS , MCM7 , RPB2 , and TEF1 , datasets were, respectively, TNe\u2009+\u2009G4, K2P\u2009+\u2009I, TNe\u2009+\u2009I and TNe\u2009+\u2009G4. All gene regions resolved the strains in two concordant well-supported clades, and with the exception of SSU, all regions resolved them in a clade separate from Cr. blascoi. Similarities between isolates from the two clades containing our isolates were low; the SSU differed by three or more substitutions, the LSU differed by eight or more substitutions, the ITS differed by 20 or more substitutions, the MCM7 differed by 99 or more substitutions, the TEF1 differed by 102 or more substitutions, and the RPB2 differed by 99 or more\u00a0substitutions.Single gene phylogenies were subsequently calculated to assess the phylogenetic relationships between our strains by applying GCPSR Fig.\u00a0. For theR. humboldtensis and distinctive characters such as the shape of the ascomata and ascospore ornamentation were observed only on branch samples, never in culture, and all strains obtained from them resided in a single clade based on DNA sequence comparisons. The ascomata on the samples emerged directly from resin or the plant tissue that surrounded it Fig.\u00a0. Ascomaties Fig.\u00a0a, b. TheRikkinen ; Table 2Cryptocalicium blascoi, the closest phylogenetic relative to our species, shares broad morphological and niche features with our fungus and R. humboldtensis . Furthermore, morphological characteristics, colony growth rate and optimal growth temperature data revealed minor but consistent differences between two species which we describe below : Conguill\u00edo National Park sector Los Paraguas: -38.697836\u00b0, -71.817216\u00b0, from ascomata on branches of A. araucana, 02 May 2018, F. Balocchi .Description: In nature \u2014 Ascomata emerging from dry resin patches in branches of A. araucana, stipitate, capitate, (556\u2013)766\u20131070(\u20131540) \u00b5m tall (excluding mazaedium); stipe cylindrical, subulate or rarely dumbbell-shaped, bent wholly or at the base, rarely straight, black, rugose, covered with a granular orange to reddish-brown mineral pruina, (257\u2013)364\u2013598(\u20131069)\u2009\u00d7\u2009(70\u2013)92\u2013214(\u2013275) \u00b5m; stipe medulla composed of textura intricata-porrecta, brownish pigmentation; stipe outermost layer composed of 15\u201320 layers of textura prismatica-porrecta, long parallel hyphae, cylindrical, septate, slightly thick walled, hyaline inner layers becoming dematiaceous towards outer layers, hyphae 2\u20133\u00a0\u00b5m wide; strongly melanized mycelium emerging from the base of the stipe, hyphae darkly pigmentated and thick walled, (3\u2013)6\u20139(\u201313.5)\u2009\u00d7\u20093\u20135(\u20137.5) \u00b5m; capitulum spherical when immature, ampulliform or urceolate when mature, (204\u2013)293\u2013394(\u2013467) \u00b5m wide, (121\u2013)230\u2013344(\u2013500) \u00b5m tall (not including mazaedium), black, rugose, sometimes covered with white to yellowish white resin; margins at the apical opening discrete, irregular, sometimes crenate-like, light brown; mazaedium well-developed in mature ascomata, reddish-brown, irregularly shaped or in some cases columnar, up to 290\u00a0\u00b5m in length; ectal excipulum poorly differentiated from the medullary excipulum, both combined consisting of 8\u201314 layers of textura prismatica\u2013porrecta, hyphae periclinally arranged, darkly pigmented in the outer layers and becoming hyaline in the inner, (8\u2013)11\u201317(\u201332)\u2009\u00d7\u20093\u20136(\u20139) \u00b5m; subhymenium hyaline or with very slight yellowish pigmentation, composed of textura angularis-intricata. Asci clavate, hyaline, pedicellate, bitunicate, evanescent, eight-spored, often with biseriately arranged spores, sporiferous part (15.5\u2013)19\u201323(\u201330)\u2009\u00d7\u20094\u20136\u00a0\u00b5m, pedicel 6\u201318.5(\u201332)\u2009\u00d7\u20091.5\u20133\u00a0\u00b5m. Ascospores non-septate, hyaline when immature, becoming brownish with age, smooth to finely roughened, broadly ellipsoidal, some globose or ellipsoidal, some erythrocyte-like, 4\u20135.5(\u20136.5)\u2009\u00d7\u2009(3.5\u2013)4\u20135.5\u00a0\u00b5m, (x\u0305\u2009=\u20095.25\u2009\u00b1\u20090.6\u2009\u00d7\u20094.34\u2009\u00b1\u20090.49), average length/width 1.22\u2009\u00b1\u20090.12 (n\u2009=\u200950). Paraphyses hyaline, filiform, obtuse, non-branched, 1\u20135 septate, 64\u2013121(\u2013172)\u2009\u00d7\u20092\u20133\u00a0\u00b5m. Phialide-like structures sporadically found inside the capitulum, potentially emerging from walls, hyaline, single-celled, subglobose, ellipsoidal or ampulliform, guttulate, (3\u2013)4.5\u20137.5(\u20138.5)\u2009\u00d7\u20092\u20133(\u20134) \u00b5m, bearing single conidia-like structures, globose to obovoid, sometimes lobsided, hyaline when immature, becoming brownish with age, smooth or finely roughened, 4\u20136(\u20137)\u2009\u00d7\u20093.5\u20135.5\u00a0\u00b5m (x\u0305\u2009=\u20095.54\u2009\u00b1\u20090.67\u2009\u00d7\u20094.47\u2009\u00b1\u20090.59), average length/width\u2009=\u20091.25\u2009\u00b1\u20090.19 (n\u2009=\u200937).In culture \u2014 colony diameters: (mm after 14 d (after 21 d)): MEA at 5\u00a0\u00b0C microcolonies (3\u20135), at 10\u00a0\u00b0C 6\u20139 (13\u201315), at 15\u00a0\u00b0C 13\u201321 (17\u201328), at 20\u00a0\u00b0C 19\u201326 (29\u201335), at 25\u00a0\u00b0C 18\u201324 (29\u201337); PDA at 5\u00a0\u00b0C microcolonies (3\u20134), at 10\u00a0\u00b0C 8\u201311 (13\u201317), at 15\u00a0\u00b0C 15\u201318 (21\u201326), at 20\u00a0\u00b0C 20\u201324 (28\u201335), at 25\u00a0\u00b0C 19\u201327 (27\u201340), at 30\u00a0\u00b0C 6\u201315 (8\u201321), at 35\u00a0\u00b0C no growth; WA at 5\u00a0\u00b0C microcolonies (2\u20135), at 10\u00a0\u00b0C 5\u20138 (10\u201313), at 15\u00a0\u00b0C 10\u201314 (15\u201320), at 20\u00a0\u00b0C 14\u201319 (22\u201327), at 25\u00a0\u00b0C 14\u201323 (25\u201336); OA at 20\u00a0\u00b0C 18\u201324 (30\u201336).Colony characters: MEA 25\u00a0\u00b0C 21 d: Colonies velvety, concentric darkening or without pattern, umbonate, margins entire to slightly irregular, subsurface, obverse greyish yellow to dark yellow (4B4\u2013C8), aerial mycelium yellowish white to orange white (4A2\u20135A2) at margins, pale to light yellow (4A3\u20135) centrally, reverse beige to blond (4C3\u20134) orange yellow to dark yellow (4B8\u2013C7) centrally. MEA 20\u00a0\u00b0C 21 d: Colonies slimy, glabrous and sulcate, rarely dry and fluffy, umbonate, with full or slightly crenate margins, obverse greyish yellow to dark yellow (4B3\u2013C8), aerial mycelium yellowish white to orange white (4A2\u20135A2) at margins, yellowish white to light yellow (4A2\u20135) centrally, reverse beige to blond (4C3\u20134) at margins, orange yellow to dark yellow (4B8\u2013C7) centrally. MEA 20\u201325\u00a0\u00b0C 6 wks or older: Colonies slimy, glabrous, sulcate, irregular margins, producing sporodochia as mucilaginous spore masses on the surface, in concentric halos or with no pattern, dark brown to brownish black (6F4\u2013H8). PDA 25\u00a0\u00b0C 21 d: Colonies velvety, glabrous, some cultures slimy only at centres, concentric or slightly petaloid pattern, umbonate, with full or slightly crenate margins, obverse yellowish grey to orange white (4B2\u20135A2), margins sunken in the medium, aerial mycelium greyish yellow to pale orange (4B4\u20135A3) at margins, becoming slightly darker or sometimes grey (4B1\u2013D1) towards the centre, reverse yellowish white to yellowish grey (4A2\u2013B2) at margins, pale yellow to olive (2D4\u20134A3) at centre, sometimes olive (2F7).Microscopic characters: Somatic hyphae at colony periphery hyaline, smooth, thin-walled, branched, transversely septate, 1.5\u20134.5\u00a0\u00b5m diam, anastomosing and developing coils, becoming inflated and melanized with age. Conidiomata most commonly absent, sporodochial with age, composed of pseudostromatic thick-walled melanized hyphae, setae absent. Conidiophores reduced, unbranched, rarely single branched or stromatic. Conidiogenous cells monophialidic or undifferentiated, then acropleurogenous; denticles sometimes present, up to 3\u00a0\u00b5m in length; terminal conidiogenous loci 13.5\u201325 (\u201335) \u00b5m in length from septa; phialides ampulliform, curved or straight and tapering towards the apex, collarette distinct, (5.5\u2013)7\u201310(\u201315)\u2009\u00d7\u20092.5\u20133(\u20134) \u00b5m. Conidia produced in slimy heads, hyaline, sometimes dematiaceous with age, aseptate, smooth walled, distinct hilum mostly present, globose to broadly ellipsoidal, larger conidia on PDA and WA typically ellipsoidal to ovoid, variable in size, from MEA (3\u2013)3.5\u20134.5(\u20136.5)\u2009\u00d7\u20093\u20133.5(\u20134) \u00b5m (x\u0305\u2009=\u20093.83\u2009\u00b1\u20090.46\u2009\u00d7\u20093.41\u2009\u00b1\u20090.26), average length/width\u2009=\u20091.13\u2009\u00b1\u20090.17 (n\u2009=\u2009228), from PDA 3.5\u20136(\u20138)\u2009\u00d7\u2009(3\u2013)3.5\u20135\u00a0\u00b5m, (x\u0305\u2009=\u20095.06\u2009\u00b1\u20091.11\u2009\u00d7\u20093.82\u2009\u00b1\u20090.46), average length/width\u2009=\u20091.32\u2009\u00b1\u20090.26 (n\u2009=\u200956), from WA (3\u2013)3.5\u20135(\u20136.5)\u2009\u00d7\u2009(2.5\u2013)3\u20133.5 (\u20134.5) \u00b5m (x\u0305\u2009=\u20094.39\u2009\u00b1\u20090.82\u2009\u00d7\u20093.3\u2009\u00b1\u20090.38), average length/width\u2009=\u20091.34\u2009\u00b1\u20090.26 (n\u2009=\u200962).Habitat/host: Resinogalea araucana produces ascomata on dry resin patches on damaged tissues in A. araucana branches. It was isolated directly from resin or plant tissues covered with resin. The type of damage inducing the exudated resin in which this fungus was found varied between samples, and included cankers caused by pathogenic fungi or insect feeding. This suggests that the presence of this fungus is merely associated with the presence of dry resin and has no role in causing resin to be produced. Strains originated from all collection sites in the Andes and Coastal range, suggesting that its distribution overlaps that of A. araucana.Material examined:Chile:Araucan\u00eda (IX): Conguill\u00edo National Park (sector Los Paraguas), -38.697836\u00b0, -71.817216\u00b0, from branches of A. araucana, 2 May 2018, F. Balocchi ; ibid, -38.697836\u00b0, -71.817216\u00b0, from branches of A. araucana (isolates from plant tissue covered in resin), 11 Dec. 2017, (culture CMW 53536\u2009=\u2009AR023); \u2014 Villarrica National Park sector Puesco, -39.572706\u00b0, -71.499235\u00b0, branches of A. araucana (isolates from plant tissue covered in resin), 13 Dec. 2017, F. Balocchi . \u2014 Biob\u00edo (VIII), Ralco Natural Reserve, -37.962620\u00b0, -71.327679\u00b0, cankers on branches on A. araucana (isolates from plant tissue covered in resin), 27 Dec. 2017, F. Balocchi (culture CMW 53543\u2009=\u2009AR224); \u2014 Nahuelbuta mountain range, Trongol Alto, -37.564893\u00b0, -73.205764\u00b0, branches of A. araucana (isolates from plant tissue covered in resin), 21 Dec. 2017, F. Balocchi ; \u2014 -37.553434\u00b0, -73.188438\u00b0, ascomata on branches of A. araucana, 5 Dec. 2019, F. Balocchi .Distinguishing characters: See below.MycoBank: MB 846746.Etymology: Latin, \u2018tapulicola\u2019, named after the Mapudungun (indigenous Chilean and Argentinian Mapuche language) word \u2018tap\u00fcl\u2019 meaning the leaves of a tree, and the Latin '-cola' meaning an inhabitant, inhabitant of leaves of a tree.Diagnosis: ITS barcode: OP508126. Alternative identification markers: LSU\u2009=\u2009OP508138, MCM7\u2009=\u2009OP524462, RPB2\u2009=\u2009OP524476, SSU\u2009=\u2009OP508114, TEF1\u2009=\u2009OP524488.Type:Chile:Araucan\u00eda (IX): Conguill\u00edo National Park sector Los Paraguas, -38.697836\u00b0, -71.817216\u00b0, branches of A. araucana (isolates from plant tissue covered in resin), 11 Dec. 2017, F. Balocchi .Description: Only known in culture. Colony diameters: (mm after 14 d (after 21 d)): MEA at 5\u00a0\u00b0C microcolonies (3\u20134), at 10\u00a0\u00b0C 9\u201310 (15\u201317), at 15\u00a0\u00b0C 15\u201318 (24\u201326), at 20\u00a0\u00b0C 22\u201327 (33\u201339), at 25\u00a0\u00b0C 23\u201331 (33\u201347); PDA at 5\u00a0\u00b0C microcolonies (3\u20135), 10\u00a0\u00b0C 9\u201312 (16\u201319), 15\u00a0\u00b0C 17\u201320 (25\u201330), 20\u00a0\u00b0C 21\u201329 (32\u201344), at 25\u00a0\u00b0C 27\u201334 (39\u201349), 30\u00a0\u00b0C 11\u201319 (15\u201327), 35\u00a0\u00b0C no growth; WA at 5\u00a0\u00b0C microcolonies (3\u20135), at 10\u00a0\u00b0C 7\u20139 (10\u201316), at 15\u00a0\u00b0C 13\u201316 (17\u201324), at 20\u00a0\u00b0C 15\u201323 (22\u201334), at 25\u00a0\u00b0C 17\u201324 (29\u201336); OA at 20\u00a0\u00b0C 20\u201324 (29\u201337).Colony characters: MEA 25\u00a0\u00b0C 21 d: Colonies velvety to tomentose, mycelium growth in a rosaceous and/or concentric pattern, umbonate, margins irregular, obverse dark yellow to greyish yellow (4B4\u2013C8) at margins, aerial mycelium yellowish white to orange grey (4A2\u20135B2) at margins, pale to light yellow (4A3\u20135) centrally, reverse greyish yellow to blond (4B3\u2013C4) at margins, becoming dark yellow to brownish yellow (4B8\u20135C7) centrally. MEA 20\u201325\u00a0\u00b0C 6 wks or older: slimy, glabrous, rarely velvety, umbonate, mycelium growing in a rosaceous and/or concentric pattern, margins irregular, soluble pigments golden brown to reddish brown (5D7\u20138E6), obverse orange grey to light brown (6B2\u2013D4), aerial mycelium sometimes present, white to greyish red (3A1\u20137B4), reverse brownish orange to brown (5C3\u2013F6), sporodochia appear as mucilaginous spore masses on the surface, in concentric halos or with no pattern, light brown to dark brown (6D6\u2013F8). PDA 25\u00a0\u00b0C 21 d: slimy, glabrous to tomentose, mycelium growing in a rosaceous or stellate pattern, margins irregular, obverse edges sunken in the medium, yellowish grey (2B2\u20134B2), aerial mycelium white to pale yellow (3A1\u20133) at margins, becoming olive to dark grey (1F1\u20133E8) centrally, reverse pale grey to yellowish grey (3B1\u20133) at margins, becoming dull yellow to olive (3B4\u2013E5) centrally, sometimes darker (3F7).Microscopic characters: Somatic hyphae at colony periphery hyaline, smooth, thin-walled, branched, transversely septate, 1.5\u20134\u00a0\u00b5m diam, anastomosing, often developing coils, becoming inflated and melanized with age. Conidiomata most commonly absent, sporodochial with age, composed of interwoven thick-walled melanized hyphae, setae absent. Conidiophores reduced, unbranched, single branched, becoming monoverticillate to richly branched. Conidiogenous cells monophialidic or undifferentiated, then acropleurogenous; denticles sometimes present, up to 4.5\u00a0\u00b5m in length; terminal conidiogenous loci (10.5\u2013)19\u201328(\u201352) \u00b5m in length from septa; phialides ampulliform, sometimes constricted at the middle, curved or straight and tapering towards the apex, with distinct collarette, (6\u2013)8\u201312(\u201316)\u2009\u00d7\u2009(2\u2013)2.5\u20133.5(\u20134) \u00b5m. Conidia produced in slimy heads, hyaline and thin-walled when young or on nutrient poor media, thick-walled and/or dematiaceous with age, aseptate, smooth walled, distinct hilum mostly present, on MEA globose to broadly ellipsoidal, rarely ellipsoidal, 3.5\u20134.5(\u20136)\u2009\u00d7\u20093\u20134.5\u00a0\u00b5m (x\u0305\u2009=\u20094.28\u2009\u00b1\u20090.49\u2009\u00d7\u20093.91\u2009\u00b1\u20090.32), average length/width\u2009=\u20091.09\u2009\u00b1\u20090.09 (n\u2009=\u2009114), on PDA and WA subglobose to ellipsoidal, rarely globose, on PDA (3.5\u2013)4\u20135(\u20136)\u2009\u00d7\u20093\u20134(\u20134.5) (x\u0305\u2009=\u20094.66\u2009\u00b1\u20090.66\u2009\u00d7\u20093.55\u2009\u00b1\u20090.38), average length/width\u2009=\u20091.32\u2009\u00b1\u20090.21 (n\u2009=\u200958), on WA (3\u2013)4\u20135(\u20137.5)\u2009\u00d7\u2009(3\u2013)3.5\u20134(\u20134.5) (x\u0305\u2009=\u20094.31\u2009\u00b1\u20090.93\u2009\u00d7\u20093.62\u2009\u00b1\u20090.38), average length/width\u2009=\u20091.19\u2009\u00b1\u20090.19 (n\u2009=\u200950).Habitat /host: A sexual morph of this fungus has not been observed, and is only known from cultures isolated from plant tissues that had resin on them. Strains originate from collection sites in the Andes and Coastal range suggesting that its distribution overlaps that of A. araucana. Its occurrence on other tree species has not been explored and a broader geographical distribution may thus exist.Material examined:Chile:Araucan\u00eda (IX): Conguill\u00edo National Park sector Los Paraguas, -38.697836\u00b0, -71.817216\u00b0, branches of A. araucana (isolates from plant tissue covered in resin), 11 Dec. 2017, F. Balocchi . \u2014 Villarrica National Park sector Puesco, -39.575582\u00b0, -71.493489\u00b0, branches of A. araucana (isolates from plant tissue covered in resin), 13 Dec. 2017, F. Balocchi ; \u2014 Biob\u00edo (VIII), Nahuelbuta mountain range, Trongol Alto, -37.564893\u00b0, -73.205764\u00b0, branches of A. araucana (isolates from plant tissue covered in resin), 15 Jan. 2019, F. Balocchi .Distinguishing characters: The general morphology of the ascomata produced by R. araucana closely resembles those of R. humboldtensis, and to a lesser extent Cr. blascoi. Ascomata were not observed for R. tapulicola. Comparing ascomata of R. araucana with those reported for R. humboldtensis 92\u2013214(\u2013275) \u00b5m vs 80\u2013140\u00a0\u00b5m), and the mineral pruina covering it was orange to darker red as oppose to yellowish brown. Ascospores of R. araucana were on average slightly larger (4\u20135.5(\u20136.5)\u2009\u00d7\u2009(3.5\u2013)4\u20135.5\u00a0\u00b5m vs (3\u2013)3.6\u20134.7(\u20135.8)\u2009\u00d7\u2009(2.7\u2013)3.3\u20134.3(\u20134.6) \u00b5m) and asci were thinner (4\u20136\u00a0\u00b5m vs 6\u20139\u00a0\u00b5m) with commonly shorter pedicels (6\u201318.5(\u201332) \u00b5m vs 22\u201343\u00a0\u00b5m). Cryptocalicium blascoi produces smaller ascomata (150\u2013360\u00a0\u00b5m vs (556\u2013)766\u20131070(\u20131540) \u00b5m), generally smaller microscopic features, and is characterized by a greenish mazaedium, contrasting with the reddish brown mazaedium of R. araucana.R. araucana and R. tapulicola resemble each other and could easily be confused. Resinogalea tapulicola grows faster than R. araucana originating from them was confirmed by sequencing. This difference in results between isolates from R. araucana and R. humboldtensis may be an outcome of different ages and/or preservation status of the specimens at their time of collection and/or processing. The erythrocyte-like shape of ascospores in R. humboldtensis observed by Rikkinen et al. alongside B. castoris with Cryptocalicium, the only other genus in the family. However, we consider Bruceomyces (B. castoris) to be morphologically sufficiently distinct from Resinogalea and Cryptocalicium to merit retention as the monotypic genus of Bruceomycetaceae. Furthermore, we emphasise that DNA sequence data will be needed to determine its relationship with other fungi.The results of our analyses using DNA sequence data generated from Cryptocalicium, Prieto et al. . As a consequence, R. humboldtensis has remained taxonomically obscure and its relationship to the specimen we found on A. araucana was initially only suspected based on the hosts and substrate. This situation has been highlighted by numerous authors who note that a large number of old fungal names remain unsequenced , had no reference DNA sequence data available and had not been classified within the ascomycetes (incertae sedis). Morphological and ecological data supports our conclusions that the fungi collected in this study on A. araucana are most closely related to R. humboldtensis,\u00a0and phylogenetic analyses showed that they reside in the recently described subclass Cryptocaliciomycetidae. The genus Resinogalea has consequently been re-classified in the Cryptocaliciaceae together with the two new species R. araucana and R. tapulicola described here from A. araucana in Chile.This study combines morphological, ecological, and phylogenetic data to describe two novel Additional file 1.\u00a0Maximum likelihood tree for the ITS gene region for 28 isolates obtained from branches of Araucaria araucana."} +{"text": "DFT analysis at M06/6-311G level was accomplished to explore the photonic behavior of PCMD1\u2013D9 compounds. Various kind of analysis like; UV\u2013Vis, density of state (DOS), natural bond orbitals (NBOs), transition density matrix (TDM) and frontier molecular orbitals (FMOs) analyses were accomplished to understand the NLO properties of said chromophores. The configuration change led to considerable charge distribution over highest occupied and lowest unoccupied molecular orbitals with minimum band difference. The energy gap trend for all the entitled compounds was observed as; PCMD8\u2009<\u2009PCMD5\u2009=\u2009PCMD9\u2009<\u2009PCMD6\u2009<\u2009PCMD7\u2009<\u2009PCMD4\u2009<\u2009PCMD3\u2009<\u2009PCMD2\u2009<\u2009PCMD1 with the least band gap of 2.048\u00a0eV in PCMD8 among all the compounds. The UV\u2013Visible spectrum of the entitled chromophores manifested high values of \u03bbmax in derivatives contrary to PCMR. Additionally, NBO findings explored effective intramolecular charge transfer and maximum energy of stabilization for PCMD8 chromophore. The highest linear polarizability (<\u03b1>) and dipole moment (\u00b5tot) values were exhibited by PCMD5 at 2.712\u2009\u00d7\u200910\u201322. and 1.995\u2009\u00d7\u200910\u201317 esu, respectively. PCMD8 push\u2013pull configured molecular entity exhibited highest first hyper-polarizability (\u03b2tot) at 4.747\u2009\u00d7\u200910\u201327 esu and second hyper-polarizability at 6.867\u2009\u00d7\u200910\u201332 esu. Overall, all the formulated chromophores exhibited significant NLO results contrary to PCMR. Hence, through this structural tailoring via various acceptors, effective NLO materials were obtained for optoelectronic applications.In opto-electronics, non-fullerene (NF) derivatives are regarded as efficient non-linear optical (NLO) materials. The present investigation was based on designing NF naphthalene-based derivatives (PCMD1\u2013D9) with D-\u03c0 One particularly fascinating aspect of NLO is Second Harmonic Generation (SHG), a process that transforms incoming light waves into waves with double their initial frequency. This phenomenon has garnered extensive attention for its practical applications, particularly in advanced technologies like photovoltaics and optoelectronics2.In the past few decades, non-linear optics (NLO) has emerged as a rapidly expanding field of scientific exploration. It delves into the intricate relationship between light interacts with matter, especially when subjected to external electric fields, this phenomenon referred to as \u2018nonlinear optical phenomena\u2019 due to the non-trivial relationship between the response of matter and the strength of the applied electric field3. Each material possesses unique attributes that may make it well-suited for one application while less relevant for others. The performance of materials becomes notably significant when they exhibit a high degree of nonlinearity, demonstrates promising potential for crystal growth, and possesses a high damage threshold, among other qualities. The continuous development of novel materials with exceptional characteristics plays a pivotal role in advancing leading-edge technologies6. The strength of optical parallelism and fast speed will progressively generate optoelectronic systems where more functions could be executed optically. Nevertheless, the technological evolution of photonics dependent on formulating novel compounds with enhanced performance7.When an electric field interacts with dielectric materials, it induces a rearrangement of the spatial distribution of electrons around the nucleus. This distortion leads to the establishment of electric dipoles within the material, resulting from these electron\u2013nucleus distortions. The choice of a suitable crystal for a specific application in nonlinear optics depends on several factors, including the nonlinear phenomenon being employed, the characteristics of the pump laser, and the desired properties of the device8. Due to D\u2013\u03c0\u2013A architecture increased conjugation and these are utilized in number of fields causing organic compound having excellent NLO properties9. Conjugated polymers are considered the most comprehensive researched materials for nonlinear optics and among the organics. Having a quality of existence of a delocalized \u03c0-electron system making it quick response giving and substantial third-order nonlinear optical characteristics11. They can also be created in many geometries, such as waveguides, films, fibers, and single crystals and they can be utilized by molecular engineering. So, polymers with the \u03c0-conjugated structures are considered as top applicants for succeeding optical photonic technologies.Different NLO substances have been attained by many scientific efforts during current years to bring out synthetic resins, molecular dyes, organic and inorganic semiconductor diodes. Low dielectric constants, low cost, high photoelectric coefficients, accessibility, the contribution of \u03c0-bonding system and electronic displacement besides facile formulation made the organic compounds should be selected in preferences. Intra-molecular charge transfer (ICT) is an important phenomenon in NLO response. The NLO substances demonstrating \u201cpush\u2013pull\u201d system due to donor-\u03c0-acceptor framework is the reason for development of ICT13, reduced ground state dipole interaction, chirality15, and organometallic complex18, however physical and chemical methods have been successful in attaining non-centro symmetric organic compounds.Due to electronic delocalization, particularly conjugation present in organic materials, they exhibit distinctive optoelectronic characteristics such as photocatalytic, photovoltaic and photoconductive behavior. High second order nonlinear optical response is present in organic compounds demonstrating intramolecular charge transfer (ICT). Charge spread asymmetrically in the p electron structure in organic materials having electron deficient (acceptor) and rich (donor) motifs and hence exhibit enhanced NLO behavior. The conditions of chemical and mechanical stability, large damage threshold, and high phase matchable NLO coefficient must be present in NLO crystals. Molecular properties based on a few of the above conditions can be fulfilled through molecular formulation. By choosing appropriate acceptor and donor combinations, the molecular ICT can be regulated. Various approaches have been presented to develop organic compounds with non-centro symmetric structures like hydrogen bonding\u03b2 values) and high nonlinear optical coefficients. Due to their expanded aromatic rings in structure, they have an extremely conjugated \u03c0-electron system. This conjugation gives rise to efficient electron delocalization and charge transfer, which are important for generating nonlinear optical responses. Having highly linear planar structure and having strong molecular packing makes the octacyclic naphthalene-based organic scaffolds (NITT) selection as \u03c0-spacer for current study. Yang et al. inserted a terminal end group i.e., IC-2F with NITT core, namely NITTBF to further explore nonlinear properties19. NITT-BF compound showed red shifted absorption along with higher electron mobility because of enhanced intermolecular \u03c0\u2013\u03c0 stacking. Its blend film gives increased exciton dissociation and charge collection properties. NITT-BF delivers higher electron mobility and stronger intermolecular \u03c0\u2013\u03c0 stacking, which account for the higher exciton dissociation and charge collection efficiency in NITT-BF-based device. In addition, NITT-BF has an optical energy gap of 1.25 eV which corresponds to higher nonlinearity.A large and delocalized \u03c0-electron system of octacyclic naphthalene-based organic compounds which leads to exceptional hyperpolarizabilities configured PCMD1\u2013D9 organic chromophores by substituting acceptor motif in the reference PCMR having acceptor\u2013\u03c0\u2013acceptor (A\u2013\u03c0\u2013A) configuration from parent NITT-BF. The reference compound has been obtained by substituting R1 (3-ethylheptane) and R2 (1-hexyl-4-methylbenzene) with methyl group to reduce computational cost and time of investigation. However, in PCMD9 the one side acceptor is replaced with donor motif which remained constant in the rest of the compounds. But from PCMD1\u2013D9 the end capped acceptor group was substituted by variant acceptor groups to study their molecular nonlinearity and impact on ICT. The analyses were performed at M06/6-311G functional. The TD-DFT and DFT calculations would be executed to interpret the effect of variant acceptors on intramolecular charge transmission, band gap, nonlinear response and absorption spectra. Therefore, various analysis such as FMO, NLO, NBO, UV\u2013Vis, TDM and DOS were performed. We hope these theoretically engineered molecules will lead to more advancements in leading optical technology.20. Complete investigation of the present research were accomplished utilizing M0621/6-311G22 theory level. Transition density matrix (TDM) as well as natural bond orbital (NBO) analyzed charge transition interactions through Multiwfn 3.723 and NBO package 3.124. The energies of frontier molecular orbitals (FMOs) and orbital diagrams were obtained through Avogadro25. The Ultraviolet\u2013Visible (UV\u2013Vis) study was executed utilizing GaussSum26 and the spectral diagram was depicted via Origin 8.027. However, the density of states illustrations were obtained through PyMOlyze28. The dipole moment (\u00b5tot)29 and nonlinear optical parameters like linear polarizability (<\u03b1>)30 as well as nonlinear hyperpolarizability (\u03b2tot and \u03b3tot)31 were also computed at the same level M06/6-311G through Eqs.\u00a0 and PCMD1\u2013D9 having donor-\u03c0-acceptor (D\u2013\u03c0\u2013A) system) were calculated employing density functional theory (DFT) via Gaussian 09 program\u03c3), chemical potential (\u03bc)32, ionization potential (IP)33, electronegativity (X)34, global hardness (\u03b7)35, electron affinity (EA) and global electrophilicity index (\u03c9)36.Equations\u00a0\u201311) wer wer11) w19 was utilized. The structural modification of NITT-BF into three parts; 2- malononitrile two acceptor groups on either side and NITT core. The R1 (3-ethylheptane) and R2 (1-hexyl-4-methylbenzene) groups in NITT core were replaced by methyl groups to convert NITT-BF to reference PCMR with A\u2013\u03c0\u2013A configuration energy gap is directly influenced by above mentioned factors. These orbitals determine intra-molecular charge transference efficiency42. However, their energy difference (\u0394E) is efficient to know molecular chemical reactivity as well as dynamic stability Less \u0394E value corresponds to high polarizability which in turn lead to exceptional NLO behavior43. On the other hand, high \u0394E value corresponds to molecular stability and hardness leading to less reactivity and chemical alteration. Table NLO characteristics of molecule\u2019s electronic structure is determine by frontier molecular investigationFrom the above table, the reference organic molecule (PCMR) possesses\u2009\u2212\u20095.941 and\u2009\u2212\u20093.505\u00a0eV of HOMO and LUMO energies, which are comparable to the experimental values of\u2009\u2212\u20095.75 and\u2009\u2212\u20094.15\u00a0eV, respectively, indicating the accurateness in the selection of functional group for the present analysis. The achieved values of HOMO for all the derivatives (PCMD1\u2013D9) are higher than the reference compound at\u2009\u2212\u20095.658,\u2009\u2212\u20095.637,\u2009\u2212\u20095.649,\u2009\u2212\u20095.657,\u2009\u2212\u20095.697,\u2009\u2212\u20095.682,\u2009\u2212\u20095.659,\u2009\u2212\u20095.696 and\u2009\u2212\u20095.693\u00a0eV, respectively. However, the obtained LUMO values for PCMD1\u2013D4 and PCMD7 are higher than the reference i.e.,\u2009\u2212\u20093.362,\u2009\u2212\u20093.352,\u2009\u2212\u20093.402,\u2009\u2212\u20093.445 and\u2009\u2212\u20093.459\u00a0eV, correspondingly but on the other hand PCMD5, PCMD6, PCMD8 and PCMD9 exhibit less values than PCMR at\u2009\u2212\u20093.623,\u2009\u2212\u20093.525, 3.648 and\u2009\u2212\u20093.619\u00a0eV, respectively. Hence, the high HOMO and less LUMO values result in less band gap in PCMD5, PCMD6, PCMD8 and PCMD9 showing high charge transmission probability in these molecules.The highest band gap is exhibited by PCMR at 2.436\u00a0eV. This largest energy difference is attributed to the presence of 2-malononitrile as a acceptor motifs with A\u2013\u03c0\u2013A configuration. Because moving from PCMR to PCMD1 the one end capped acceptor group is substituted with 9-phenyl-9H-carbazole which significantly reduces the band gap to 2.296\u00a0eV in PCMD1.Conversely, the least band gap value of 2.048\u00a0eV is present in PCMD8, that might be owing to the presence of high electronegative nitro functional groups at 6 and 7 positioning of 1-(dicyanomethylene)-3-oxo-2,3-dihydro-1H-cyclopenta[b]naphthalene (DMP) acceptor group. The second lowest energy gap value is exhibited by PCMD5 and PCMD9 at 2.074\u00a0eV because of the presence of sulfonic acid groups and cyano groups at 6 and 7 positioning of DMP acceptor moiety rendering high electron withdrawing tendency. However, PCMD6 possess 2.157\u00a0eV energy gap resulted from the 6, 7 positioning of trifluoromethyl functional group in DMP. The presence of acetate, chloro and fluoro groups at 6, 7 positioning of DMP acceptor group in PCMD7, PCMD4 and PCMD3 corresponds to band gap of 2.200, 2.212 and 2.247\u00a0eV, respectively. 2.285\u00a0eV of energy gap is present in PCMD2 in relation to the absence of any electron withdrawing functional groups in DMP. The ascending order of energy difference (HOMO/LUMO) for the derivatives is PCMD8\u2009<\u2009PCMD5\u2009=\u2009PCMD9\u2009<\u2009PCMD6\u2009<\u2009PCMD7\u2009<\u2009PCMD4\u2009<\u2009PCMD3\u2009<\u2009PCMD2\u2009<\u2009PCMD1. Figure\u00a044. In DOS pictographs, the HOMO signifies valence band showing negative values while the positive values are represented by the conduction band (LUMO)45. The analysis was performed at the same DFT functional for\u00a0PCMR and PCMD1\u2013D9. DOS indicates the contribution of each fragment of molecule in charge transfer. For determining the density of states (DOS), we split our studied molecules into se98parate fragments. The\u00a0PCMR1 was divided into two fragments\u00a0i.e., donor (D) and acceptor (A) while, the derivatives (PCMD1\u2013D9) were divided into three segments\u00a0i.e., donor, \u03c0-spacer, and acceptor. From Fig.\u00a0The DOS analysis verify the results shown by the FMOs diagrams and explain the electronic distribution in frontier molecular orbitals47. Franck\u2013Condon principle relates spectral highest absorption maxima (\u03bbmax) to vertical excitation. The information about oscillator strength (fos), absorption maxima (\u03bbmax), excitation energy (E), along with molecular orbital (MO) contribution in transition are given in Table \u03bbmax) in ultraviolet visible region for PCMR and PCMD1\u2013D9 is presented in Fig.\u00a0The ultraviolet visible computation in gaseous form for PCMR and PCMD1\u2013D9 were performed at M06/6-311G to apprehend oscillator strength as well as excitation energy with relation to important electronic transitions\u03bbmax) value for reference (PCMR) i.e., 606.899\u00a0nm is lowest among all the compounds with transition energy of 2.043\u00a0eV, 2.956 oscillator strength and molecular orbital contribution from HOMO to LUMO of 95%, however, from HOMO-1 to LUMO-1 it is 3%. Hence, it is observed that absorption maxima are significantly impacted by highly electron rich donor and electron withdrawing acceptor end capped groups as well as molecular configuration (reference (A\u2013\u03c0\u2013A)\u03bbmax value of reference compound resulted from two electron withdrawing acceptor groups on either side of the \u03c0-linker. However, the highest \u03bbmax value of 701.541\u00a0nm is exhibited by PCMD8 among all the chromophores because of high electronegative nitro functional groups present in DMP acceptor group, results in 1.767\u00a0eV energy of transition and oscillator strength of 1.299. The molecular orbital contribution exhibited by PCMD8 compound is 96% from HOMO to LUMO and 3% HOMO-1 to LUMO. The second highest \u03bbmax of 696.849\u00a0nm is present in PCMD5 due to the presence of sulfonic acid groups in DMP with 1.779\u00a0eV of transition energy, 1.779 fos and MO contribution of 95 and 3% from HOMO to LUMO and HOMO-1 to LUMO, respectively. However, the third highest \u03bbmax of 695.442\u00a0nm is shown by PCMD9 because of cyano groups presence in DMP with transition energy and fos values of 1.783\u00a0eV and 1.362, respectively. However, the molecular orbital contributions in PCMD9 corresponds to 95% from HOMO to LUMO and 3% from HOMO-1 to LUMO. Owing to the presence of trifluoromethyl groups in PCMD6, the \u03bbmax reduces to 663.580\u00a0nm with E 1.868\u00a0eV, 1.504 fos and MO contributions of 95 and 3% from HOMO to LUMO and HOMO-1 to LUMO, respectively. Substituting trifluoromethyl groups by chloro groups result in \u03bbmax of 641.238\u00a0nm in PCMD4 with E 1.934\u00a0eV, 1.687 fos and MO contributions of 95 and 3% from HOMO to LUMO and HOMO-1 to LUMO, respectively. The overall decreasing trend for PCMD1\u2013D9 as; PCMD8\u2009>\u2009PCMD5\u2009>\u2009PCMD9\u2009>\u2009PCMD6\u2009>\u2009PCMD4\u2009>\u2009\u2009>\u2009PCMD3\u2009>\u2009PCMD1\u2009>\u2009PCMD2 with chief molecular orbital contribution is from HOMO to LUMO of 94\u201396%. The compounds with lower Eg showed wider absorption spectrum as the decreasing trend of \u03bbmax is almost similar with the increasing trend of energy gap. All fabricated molecules showed bathochromic shift 49. \u03c3\u2009\u2192\u2009\u03c3*, \u03c0\u2009\u2192\u2009\u03c0*, LP\u2009\u2192\u2009\u03c3* and LP\u2009\u2192\u2009\u03c0* orbital overlapping lead to hyper conjugation. The \u03c0-conjugation present in the computed derivatives having D\u2013\u03c0\u2013A configuration lead to \u03c0\u2009\u2192\u2009\u03c0* transitions, hence result in efficient nonlinear optical substances. Weak \u03c3\u2009\u2192\u2009\u03c3* transitions are also present because of poor donor and acceptor interactions. Table Here, Molecular orbitals for PCMR and PCMD1\u2013D9 involve in consistent \u03c0\u2009\u2192\u2009\u03c0* transitions are \u03c0(C18\u2013C19)\u2009\u2192\u2009\u03c0*(C22\u2013C23) corresponds to stabilization energies of 31.57, 32.39, 31.82, 32.24, 32.6, 34.13, 33.46, 32.84, 34.31 and 34.06\u00a0kcal/mol, respectively. The lowest \u03c0\u2009\u2192\u2009\u03c0* transitions exhibited by PCMR and PCMD1\u2013D9 corresponds to \u03c0(C61\u2013C62)\u2009\u2192\u2009\u03c0*(C61\u2013C62), \u03c0(C34\u2013O35)\u2009\u2192\u2009\u03c0*(C28\u2013C29), \u03c0(C107\u2013C108)\u2009\u2192\u2009\u03c0*(C107\u2013C108), \u03c0(C80\u2013C83)\u2009\u2192\u2009\u03c0*(C80\u2013C83), \u03c0(C105\u2013C106)\u2009\u2192\u2009\u03c0*(C105\u2013C106), \u03c0(C39\u2013N40)\u2009\u2192\u2009\u03c0*(C37\u2013N38), \u03c0(C95\u2013C96)\u2009\u2192\u2009\u03c0*(C95\u2013C96), \u03c0(C95\u2013C96)\u2009\u2192\u2009\u03c0*(C95\u2013C96), \u03c0(C39\u2013N40)\u2009\u2192\u2009\u03c0*(C37\u2013N38) and \u03c0(C37\u2013N38)\u2009\u2192\u2009\u03c0*(C39\u2013N40) at 0.65, 4.16, 0.67, 0.51, 0.67, 0.8, 0.67, 0.67, 0.81 and 0.79\u00a0kcal/mol stabilization energy, respectively.Likewise, orbitals involved in \u03c3\u2009\u2192\u2009\u03c3* transitions in PCMR and PCMD1\u2013D9 are \u03c3(C22\u2013H24)\u2009\u2192\u2009\u03c3*(C18\u2013S20), \u03c3(C22\u2013H24)\u2009\u2192\u2009\u03c3*(C18\u2013S20), \u03c3(C22\u2013H24)\u2009\u2192\u2009\u03c3*(C18\u2013S20), \u03c3(C22\u2013H24)\u2009\u2192\u2009\u03c3*(C18\u2013S20), \u03c3(C22\u2013H24)\u2009\u2192\u2009\u03c3*(C18\u2013S20), \u03c3(C12\u2013C13)\u2009\u2192\u2009\u03c3*(C13\u2013C19), \u03c3(C3\u2013C4)\u2009\u2192\u2009\u03c3*(C4\u2013C5), \u03c3(C22\u2013H24)\u2009\u2192\u2009\u03c3*(C18\u2013S20), \u03c3(C3\u2013C4)\u2009\u2192\u2009\u03c3*(C4\u2013C5) and \u03c3(C62\u2013C64)\u2009\u2192\u2009\u03c3*(C66\u2013C84) at highest energy of stabilization of 10.27, 10.36, 10.42, 10.43, 10.48, 4.92, 4.99, 10.54, 4.99 and 4.99\u00a0kcal/mol, respectively. Whereas, the lowest stabilization energy for \u03c3\u2009\u2192\u2009\u03c3* transitions involve \u03c3(C37\u2013N38)\u2009\u2192\u2009\u03c3*(C33\u2013C36), \u03c3(C65\u2013C66)\u2009\u2192\u2009\u03c3*(C6\u2013C15), \u03c3(C62\u2013C64)\u2009\u2192\u2009\u03c3*(C62\u2013S63), \u03c3(C61\u2013C62)\u2009\u2192\u2009\u03c3*(C6\u2013C59), \u03c3(C61\u2013C62)\u2009\u2192\u2009\u03c3*(C6\u2013C59), \u03c3(C62\u2013C64)\u2009\u2192\u2009\u03c3*(C62\u2013S63), \u03c3(C62\u2013C64)\u2009\u2192\u2009\u03c3*(C62\u2013S63), \u03c3(C61\u2013C62)\u2009\u2192\u2009\u03c3*(C6\u2013C59), \u03c3(C37\u2013N38)\u2009\u2192\u2009\u03c3*(C33\u2013C36) and \u03c3(C61\u2013C62)\u2009\u2192\u2009\u03c3*(C6\u2013C59) at 0.51\u00a0kcal/mol in PCMR and 0.5\u00a0kcal/mol in PCMD1\u2013D9 chromophores.LP\u2009\u2192\u2009\u03c0* and LP\u2009\u2192\u2009\u03c3* transitions are also observed conforming to resonance. LP\u2009\u2192\u2009\u03c0* transitions involve LP(1)(C68)\u2009\u2192\u2009\u03c0*(C64\u2013C66), LP(1)(N82)\u2009\u2192\u2009\u03c0*(C83\u2013C85), LP(1)(N96)\u2009\u2192\u2009\u03c0*(C107\u2013C108), LP(2)(S20)\u2009\u2192\u2009\u03c0*(C12\u2013C13), LP(2)(S20)\u2009\u2192\u2009\u03c0*(C12\u2013C13), LP(1)(C3)\u2009\u2192\u2009\u03c0*(C9\u2013C57), LP(1)(N94)\u2009\u2192\u2009\u03c0*(C95\u2013C96), LP(2)(O120)\u2009\u2192\u2009\u03c0*(C118\u2013O119), LP(3)(O117)\u2009\u2192\u2009\u03c0*(N115\u2013O116) and LP(1)(N94)\u2009\u2192\u2009\u03c0*(C105\u2013C106) orbitals having 72.81, 35.44, 35.42, 29.54, 29.56, 69.77, 35.34, 47.48, 194.76 and 35.4\u00a0kcal/mol in PCMR and PCMD1\u2013D9 chromophores, correspondingly. All the same, LP(1)(O35)\u2009\u2192\u2009\u03c3*(C29\u2013C34), LP(1)(O35)\u2009\u2192\u2009\u03c3*(C29\u2013C34), LP(1)(O35)\u2009\u2192\u2009\u03c3*(C29\u2013C34), LP(1)(O35)\u2009\u2192\u2009\u03c3*(C23\u2013C34), LP(3)(O119)\u2009\u2192\u2009\u03c3*(S118\u2013O122), LP(1)(O35)\u2009\u2192\u2009\u03c3*(C29\u2013C34), LP(1)(O116)\u2009\u2192\u2009\u03c3*(C115\u2013O117), LP(1)(O35)\u2009\u2192\u2009\u03c3*(C29\u2013C34) and LP(1)(O35)\u2009\u2192\u2009\u03c3*(C29\u2013C34) at 21.25, 21.3, 20.67, 20.8, 18.57, 29.05, 21.11, 33.38, 21.35 and 21.24\u00a0kcal/mol in PCMR and PCMD1\u2013D9 chromophores, correspondingly. Hence, this analysis reveals that their stabilization is significantly influenced by extended hyperconjugation and strong intramolecular charge transfer. These factors underscore the importance of their charge transfer properties, which are pivotal for their nonlinear optical (NLO) characteristics.\u03c3), chemical potential (\u03bc)32, ionization potential (IP)33, electronegativity (X)34, global hardness (\u03b7)35, electron affinity (EA) and global electrophilicity index (\u03c9)36 that impart knowledge about molecular stability and its reactivity. Ionization potential (IP) is the principal parameter which defines energy needed to eliminate electron out of MO. From Table IP value is present in PCMD1 i.e. 5.64\u00a0eV. The trend for IP value is; PCMD9\u2009>\u2009PCMD4\u2009=\u2009PCMD7\u2009>\u2009PCMD8\u2009>\u2009PCMD5\u2009>\u2009PCMD3\u2009=\u2009PCMR\u2009>\u2009PCMD2\u2009>\u2009PCMD1. Conversely, energy liberates on electronic insertion in valence shell is termed as electron affinity (EA). The highest EA is exhibited by PCMD7 at 3.65\u00a0eV while 3.35\u00a0eV is the least energy released upon electronic addition in PCMD1. Knowledge about polarization of electronic cloud is collected from global softness (\u03c3) and global hardness (\u03b7) values. High polarization of electronic cloud is demonstrated by softer molecules while least is associated with harder compounds. The highest \u03b7 value is present in PCMD9 at 1.22\u00a0eV that reduces to 1.02\u00a0eV in PCMD7. The values of global softness (\u03c3) are lesser than their respective global hardness (\u03b7) and global electrophilicity index (\u03c9) values. PCMD7 manifests highest softness at 4.88\u00a0eV\u22121 with 0.436\u00a0eV\u22121 being the least softness among all compounds present in PCMD1. Hence all the fabricated chromophores exhibited the higher value of softness which supported the greater charge transference in them than reference chromophore which in result would express good NLO response. The highest value of global electrophilicity index (\u03c9) is possessed by PCMD4 at 10.47\u00a0eV. While the lowest of 8.86\u00a0eV is present in PCMR. Chemical potential (\u03bc) is inversely related to Electronegativity (X) value. Where, chemical potential describes affinity of electron removal while electronegativity reveals affinity to accept electrons. PCMD9 with highest electronegativity and lowest chemical potential at 4.72 and\u2009\u2212\u20094.72\u00a0eV, correspondingly, manifests highest electron transmission in turn possess highest nonlinearity.Global reactivity parameters (GRPs) encompass multiple descriptors i.e. global softness demonstrate density of transition which is the estimation of excitations involving charge transmission. Whereas, local excitations corresponds to transition density demonstrated by equivalent basis functions (correspondent to same atomic centers)51.Transition density matrices (TDMs) is employed for visualizing and investigating optical nature of each electronic transition in a molecular system during excited state charge dispersion and electron\u2013hole mobilityThe current inquiry of PCMR and PCMD1\u2013D9 has been executed at M06/6-311G via Multiwfn 3.7 . While atomic and molecular nonlinearity subject to extensive nonlinear optical phenomenon is hyperpolarizability . The first order hyperpolarizability (\u03b2tot), second order hyperpolarizability (\u03b3tot), linear polarizability (<\u03b1>) and dipole moment (\u00b5tot) values including tensors for PCMR and PCMD1\u2013D9 are presented in Tables NLO is an eminent discipline of current investigations due to its importance in managing primary tasks of optical memory, telecommunications, signal processing, frequency shifting, optical interconnections, optical logic, optical switching and optical modulation\u00b5) is greatly affected by difference of electronegativity that could be categorically the product of charge magnitude and distance among them, where high electronegativity corresponds to large \u00b555. Besides, molecular \u00b5, polarity is crucial in improving molecular nonlinearity. The \u00b5tot characterize average dipole moment, whereas, tensors contributes to \u00b5tot along x, y and z-orientations are \u00b5x, \u00b5y and \u00b5z56. The highest \u00b5tot values are detected in all the derivatives (PCMD1\u2013D9) in 0.981\u20131.995\u2009\u00d7\u200910\u201317 esu range contrary to reference (PCMR) at 0.308\u2009\u00d7\u200910\u201317esu. The descending order of \u00b5tot is; PCMD5\u2009>\u2009PCMD8\u2009=\u2009PCMD9\u2009>\u2009PCMD6\u2009>\u2009PCMD7\u2009>\u2009PCMD4\u2009>\u2009PCMD1\u2009>\u2009PCMD3\u2009>\u2009PCMD2\u2009>\u2009PCMR. The findings indicate major contribution by x-axis in the overall \u00b5tot 57, a standard molecule for investigating the NLO properties, these compounds exhibit 0.06, 0.23, 0.20, 0.22, 0.24, 0.40, 0.29, 0.24, and 0.37 times greater \u00b5 values. Similarly, when compared to CPTR158, similar analog to our compounds, 0.8, 3.0, 2.6, 3.0, 3.2, 5.4, 3.8, 3.2, and 5.0 the times greater values are found in our fabricated chromophores.The dipole polarizability (ot Table . The PCM\u03b1>) values of PCMR and PCMD1\u2013D9 along with polarizability contributing tensors and their values. The\u2009<\u03b1>\u2009value of 2.712\u2009\u00d7\u200910\u201322 esu is the highest polarizability value among all the chromophores exhibited by PCMD5. However, the least\u2009<\u03b1>\u2009value of 2.310\u2009\u00d7\u200910\u201322 esu corresponds to PCMD1. The\u2009<\u03b1>\u2009values for the rest of the compounds PCMR, PCMD2-D4, PCMD6-D9 are 2.499, 2.495, 2.506, 2.608, 2.588, 2.670, 2.686 and 2.695\u2009\u00d7\u200910\u201322 esu, respectively. The investigation extends to the realm of linear polarizability, where a comparison is drawn between our computed compounds (PCMR and PCMD1\u2013D9) and the standard pNA57 and CPTR158. Notably, the linear polarizability values of pNA57 and CPTR158 stand at 1.178\u2009\u00d7\u200910\u201323 and 1.370\u2009\u00d7\u200910\u201322 esu, respectively. In contrast, our designed derivatives exhibit intriguing linear polarizability values in relation to pNA and CPTR1. Correspondingly, the comprehensive analysis of their contributing tensors indicates major contribution by \u03b1xx towards the overall\u2009<\u03b1>\u2009in all the compounds. The formulated chromophores have shown comparable\u2009<\u03b1>\u2009values with given descending order; PCMD5\u2009>\u2009PCMD9\u2009>\u2009PCMD8\u2009>\u2009PCMD7\u2009>\u2009PCMD4\u2009>\u2009PCMD6\u2009>\u2009PCMD3\u2009>\u2009PCMR\u2009>\u2009PCMD2\u2009>\u2009PCMD1.Table \u03b2tot) value which increases in strongly push\u2013pull configured molecular systems because of extended conjugation. The nine tensors i.e., \u03b2xxx,\u00a0\u03b2xxy,\u00a0\u03b2xyy,\u00a0\u03b2yyy,\u00a0\u03b2xxz,\u00a0\u03b2yyz,\u00a0\u03b2xzz,\u00a0\u03b2yzz,\u00a0\u03b2zzz are used to determine hyperpolarizability values. Table \u03b2tot) values are tabulated in Table \u03b2tot highest of 4.747\u2009\u00d7\u200910\u201327 esu is exhibited by PCMD8, whereas the least hyperpolarizability is present in PCMR at 0.099\u2009\u00d7\u200910\u201327 esu. The same trend for second hyperpolarizability values has been observed, where the highest \u03b3tot of 6.867\u2009\u00d7\u200910\u201332 esu is exhibited by PCMD8, but the least \u03b3tot is present in PCMD1 at 3.517\u2009\u00d7\u200910\u201332 esu. Where the major contribution towards \u03b2tot is exhibited by \u03b2xxx tensor and \u03b3tot in is by \u03b3x tensor. Comparative analysis with pNA and CPTR1 our compounds illustrated higher values for \u03b2 and \u03b3tot50. In conclusion, the changing configuration from A-D-A to D\u2013\u03c0\u2013A and variant acceptor groups indicates substantial ICT besides bathochromic shift which in turn enhances the optical nonlinearity in all the designed chromophores. The high values of hyperpolarizability and polarizability manifest designed compounds as potential candidate for technologically advanced optical devices.Compound\u2019s nonlinearity is calculated by first hyper-polarizability with greater red shift (701.541\u00a0nm) is examined in PCMD8 than that of other molecules. From all the devised NLO compounds, PCMD8 demonstrated relatively high first hyperpolarizability and second hyperpolarizability of 4.747\u2009\u00d7\u200910\u201327 esu and 6.867\u2009\u00d7\u200910\u201332 esu, respectively. Nevertheless, PCMD5 manifested high dipole moment and linear polarizability value of 1.995\u2009\u00d7\u200910\u201317 esu and 2.712\u2009\u00d7\u200910\u201322 esu, respectively. So, PCMD8 and PCMD5 possessed high optoelectronic (linear and nonlinear) behavior resulted from the induction of nitro and sulfonic acid groups in the acceptor moiety. However, all the formulated compounds have shown higher nonlinearity on comparison with pNA as a standard molecule from the literature. Subsequently, PCMD8 chromophores is approved to be proficient NLO candidates for technologically advanced nonlinear optical devices.In current report, we devised novel octacyclic naphthalene-based PCMD1\u2013D9 nonlinear organic compounds by substituting one end acceptor moiety in PCMR with donor (9-phenyl-9H-carbazole) in derivatives and altering acceptor group in each successive derivative. Quantum chemical calculation were applied to investigate the NLO behavior of fabricated chromophores. The NBO study revealed a hyper conjugative interaction played significant role in stabilizing the molecule. An efficient charge transference from donor to acceptor through spacer has been studied by FMO findings which were further also supported by DOS and TDM analysis. A lower band gap (2.048\u00a0eVSupplementary Information."} +{"text": "N,N\u2032-bidentate ligands [4-(2-amino\u00adeth\u00adyl)morpholine] and two trans-located acetate mol\u00adecules. The Cd atom is located on a centre of inversion, whereas the morpholine and four water mol\u00adecules are adjacent to the acetate moieties. In the crystal, neighboring metal complexes and uncoordinated water mol\u00adecules are linked via N\u2014H\u22efO and O\u2014H\u22efO hydrogen-bonding inter\u00adactions.In the title coordination compound, the Cd atom is octa\u00adhedrally coordinated by two 2H3O2)2(C6H14N2O)2]\u00b74H2O, was synthesized by mixing 2 moles of 4-(2-amino\u00adeth\u00adyl)morpholine and 1 mole of cadmium acetate in double-distilled water. The Cd atom is octa\u00adhedrally coord\u00adinated by two N,N\u2032-bidentate ligands [4-(2-amino\u00adeth\u00adyl)morpholine] and two trans-located acetate mol\u00adecules. The Cd atom is located on a center of inversion, whereas the 4-(2-amino\u00adeth\u00adyl)morpholine and four water mol\u00adecules are adjacent to the acetate mol\u00adecules. The chair conformation of the morpholine mol\u00adecules is confirmed. In the crystal, adjacent metal complexes and uncoord\u00adinated water mol\u00adecules are linked via N\u2014H\u22efO and O\u2014H\u22efO hydrogen-bonding inter\u00adactions, generating R22(6), R66(16), R66(20) and S11(6) motifs and forming a three-dimensional network. A Hirshfeld surface analysis indicated the contributions of various contacts: H\u22efH (71.8%), O\u22efH/H\u22efO (27.1%), and C\u22efH/H\u22efC (1.0%).The title coordination compound, [Cd(C The coordination polyhedron around the metal atom may be best described as a distorted octa\u00adhedron. The four nitro\u00adgen atoms of the di\u00adamine ligands define the equatorial plane, and two oxygen atoms from the acetate anions coordinate in the trans-axial positions. The coordination of the morpholine ligands creates two five-membered chelate rings by varying the M\u2014N distances and N\u2014M\u2014N angles. Many articles and reviews have reported that an important factor for metal-ion selection is the chelate ring size, in which five-membered chelate rings promote selectivity for large metal ions with an ionic radius (r+) close to 1.0\u2005\u00c5. Theoretical calculations show that for five-membered N\u2013C\u2013C\u2013N\u2013M chelate rings, the ideal values for the N\u2014M distance and N\u2014M\u2014N angle are 2.5\u2005\u00c5 and 69\u00b0, respectively and 2.2788\u2005(15)\u2005\u00c5 (Cd\u2014N2 and Cd\u2014N2i), are in good agreement with the values reported in the literature \u2212x\u00a0+\u00a01, \u2212y, \u2212z\u00a0+\u00a01] and the cis-angles of the octa\u00adhedron involving O2 and O2i are close to the ideal value of 90\u00b0. The morpholine rings adopt a chair conformation. The acetate group is disordered over two positions of equal occupancy and in both of the crystallographically independent water mol\u00adecules, one of the protons is equally disordered over two positions. Finally, water atom O5 from the water mol\u00adecules is disordered over two positions in a 75\u2005(3):25\u2005(3) ratio.The title compound Fig.\u00a01 crystallgs Fig.\u00a02. Upon co3.et al., 1995et al., 2000a-axis direction indicated by cyan dashed lines. In the crystal, the mol\u00adecules are linked by numerous N\u2014H\u22efO and O\u2014H\u22efO inter\u00adactions are the most significant, contributing 71.8% to the total crystal packing. This major contribution may be due to van der Waals inter\u00adactions . Fig.\u00a09d shows the C\u22efH/H\u22efC inter\u00adactions, which contribute 1.0% to the Hirshfeld surface.A Hirshfeld surface analysis was performed for the complex alone (excluding the water molecules) and the two-dimensional (2D) fingerprint plots were created with 4.et al., 2016catena-[bis\u00ad-[4-(2-amino\u00adeth\u00adyl)morpholine]]\u00adnickel(II) morpholine]\u00adbis\u00ad(nitrito)nickel(II) bis\u00adnickel(II) copper(II) monohydrate bis\u00ad[4-(2-amino\u00adeth\u00adyl)morpholine]\u00addi\u00adcop\u00adper(II) morpholine-2\u03ba-N,N\u2032]nickel(II) dichloride morpholine.A search in the Cambridge Structural Database : 3301 , 2887 , 1549 , 1411 , 1342 , 1192 , 960 , 653 and 594 .As shown in the reaction scheme Fig.\u00a010, the tit6.Uiso(H) = 1.2\u20131.5Ueq(C), while the N\u2014H and O\u2014H protons were located in residual electron-density maps and refined with distance restraints (DFIX and SADI) and with Uiso(H) = 1.2Ueq(N) and 1.5Ueq(O). The acetate group was refined as disordered over two positions (ratio 50:50%) with distance, geometry and ijU restraints . H4 and H6 are disordered over two positions in a 50:50 ratio due to symmetry-related hydrogen bonds. O5 is disordered over two positions in a 75\u2005(3):25\u2005(3) ratio. As both positions have the same distance to H5, H6 and H6\u2032, only one set of the hydrogen atoms was refined for both O5 and O5B.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023008782/jq2030sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023008782/jq2030Isup4.hklStructure factors: contains datablock(s) I. DOI: 2299387CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules with the same absolute configuration are linked into infinite chains along the b-axis direction by O\u2014H\u22efO hydrogen bonds between the hy\u00addroxy substituent and the carbonyl O atom of the adjacent mol\u00adecule.The title compound,which is produced by the oxidation of 1-(4- 5H5)(C21H24NO2)], which is produced by the oxidation of 1-(4-tert-butyl\u00adphen\u00adyl)-2-ethyl-3-ferrocenyl\u00adpyrrole, crystallizes as a racemic mixture in the centrosymmetric space group P21/n. The central heterocyclic pyrrole ring system subtends dihedral angles of 13.7\u2005(2)\u00b0 with respect to the attached cyclo\u00adpenta\u00addienyl ring and of 43.6\u2005(7)\u00b0 with the major component of the disordered phenyl group bound to the N atom. The 4-tert-butyl\u00adphenyl group, as well as the non-substituted Cp ring are disordered with s.o.f. values of 0.589\u2005(16) and 0.411\u2005(16), respectively. In the crystal, mol\u00adecules with the same absolute configuration are linked into infinite chains along the b-axis direction by O\u2014H\u22efO hydrogen bonds between the hy\u00addroxy substituent and the carbonyl O atom of the adjacent mol\u00adecule.The title compound, [Fe(C Therefore, a new centre of chirality is created at C1, which used to be an sp2 carbon atom in the starting compound. Since no chiral reaction conditions were applied, a racemate of the title compound is produced. The title compound also crystallizes as a racemic mixture in the centrosymmetric space group P21/n. The mol\u00adecular structure of the S-enanti\u00adomer is shown in Fig.\u00a01tert-butyl\u00adphenyl group, as well as the non-substituted Cp ring, are disordered with s.o.f. values of 0.589\u2005(16) and 0.411\u2005(16). Bond lengths and angles are of expected values with the C2\u2014C3 bond length of 1.336\u2005(5)\u2005\u00c5, clearly indicating a double bond. In addition, the N1\u2014C4 bond [1.366\u2005(5)\u2005\u00c5] is shortened with respect to the other nitro\u00adgen carbon bonds, as is typical for amides.The title compound, 3.b-axis direction by O\u2014H\u22efO hydrogen bonds of the C(6) type -one CSD -2-ethyl-3-ferrocenyl\u00adpyrrole were treated with 5\u2005mol% p-toluene sulfonic acid and were dissolved in 1.0\u2005mL of anhydrous ethanol. The solution was placed in a 10\u2005mL screw-cap vessel closed with parafilm. The process of the oxidation reaction was followed by thin layer chromatography and it could be observed that the reaction was finished after approximately 8 days. The reaction mixture was transferred to a Schlenk tube, the solvent was removed in vacuo and the remaining oily residue was purified by column chromatography using CH2Cl2 as the eluent. Slow evaporation of the solvent at ambient temperature led to the formation of crystalline material of the title compound . 1H NMR : (ppm) = 0.55 ; 1.31 ; 1.92 ; 2.84 ; 4.17 ; 4.44\u20134.50 ; 4.72\u20134.73 ; 6.24 ; 7.37\u20137.43 ; 7.48\u20137.52 . 13C NMR : (ppm) = 7.80 (CH3); 26.37 (CH2); 31.32 (CH3); 34.50 (C); 68.03 (CpR); 68.85 (CpR); 70.03 (Cp); 72.96 (CpR); 95.55 (C); 118.48 (=CH); 125.44 (CHPh); 125.86 (CHPh); 135.19 (CPh); 149.21 (CPh); 160.63 (C); 169.10 (C=O). MS (DEI): m/z (%) = 443\u2005(96) [M+]; 427\u2005(76) [M+\u00a0\u2212\u00a0O]; 426\u2005(40) [M+\u00a0\u2212\u00a0OH]; 425\u2005(75) [M+\u00a0\u2212\u00a0H2O]; 398\u2005(22) [M+ \u2212 3CH3]; 360\u2005(98) [M+\u00a0\u2212\u00a0C5H5\u00a0\u2212\u00a0H2O]; 322\u2005(48) [M+\u00a0\u2212\u00a0C5H5Fe]; 305\u2005(58) [M+\u00a0\u2212\u00a0C5H5Fe\u00a0\u2212\u00a0OH]; 294\u2005(64) [M+\u00a0\u2212\u00a0C5H5Fe\u00a0\u2212\u00a0CO].0.5\u2005mmol (200\u2005mg) of 1-(4-6.O2) was located in a difference-Fourier map and refined freely. All carbon-bound hydrogen atoms were placed in idealized positions and refined using a riding model with isotropic displacement parameters Uiso(H) = 1.2eqU(C) for methyl\u00adene and aromatic hydrogen atoms and H3 and Uiso(H) = 1.5eqU(C) for methyl groups. The p-tBuC6H4 and Cp groups are disordered over two positions and were found to refine well with only one free variable. The proportion of the two positions is 58.94:41.06%. SIMU, RIGU, SAME, SADI and FLAT instructions were used to restrain the geometry and displacement parameters of the disordered moieties.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023001858/jq2026sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023001858/jq2026Isup2.hklStructure factors: contains datablock(s) I. DOI: 961575CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-1,3-benzo\u00addiazol-2(3H)-one (I and II) exhibit identical bond distances and angles except for the C\u2014N\u2014C\u2014C torsion angle between the benzimidazolone backbone and the phenyl substituent, which has an effect on the crystal packing and supra\u00admolecular features. The structure of I contains a stronger C=O\u22efH\u2014N hydrogen-bonding inter\u00adaction and a weaker \u03c0\u2013\u03c0 inter\u00adaction between adjacent bezimidazolone moieties in comparison to II.The two polymorphic structures of 3-phenyl-1 I and II) of 3-phenyl-1H-1,3-benzo\u00addiazol-2(3H)-one, C13H10N2O, acquired from pentane diffusion into the solution in THF, are reported. The structures show negligible differences in bond distances and angles, but the C\u2014N\u2014C\u2014C torsion angles between the backbone and the phenyl substituent, 123.02\u2005(15)\u00b0 for I and 137.18\u2005(11)\u00b0 for II, are different. Compound I features a stronger C=O\u22efH\u2014N hydrogen bond than that in II, while the structure of II exhibits a stronger \u03c0\u2013\u03c0 inter\u00adaction than in I, as confirmed by the shorter inter\u00adcentroid distance [3.3257\u2005(8)\u2005\u00c5 in II in comparison to 3.6862\u2005(7)\u2005\u00c5 in I]. Overall, the supra\u00admolecular inter\u00adactions of I and II are distinct, presumably originating from the variation in the dihedral angle.The polymorphic structures ( The compound was prepared following the reported procedure using 1,1\u2032-carbonyl\u00addiimidazole and N1-phenyl\u00adbenzene-1,2-di\u00adamine in CH2Cl2 and blocks (II).Here we report two polymorphic structures of 3-phenyl-12.I) and blocks (II) in space groups C2/c and Pbca, respectively. The two polymorphic structures exhibit identical bond distances and angles, except for the dihedral angle of the phenyl substituent and \u2212137.18\u2005(12)\u00b0 for I and II, respectively. No additional differences are observed from an analysis of bond distances and angles.The title compounds crystallized as colorless needles \u2005\u00c5, while the corres\u00adponding distance in I is more elongated at 3.6862\u2005(7)\u2005\u00c5.Initial investigations of supra\u00admolecular features for y Figs. 2 and 3 \u25b8.I indicate the inter\u00admolecular inter\u00adactions of C4\u22efC2/C2\u22efC4, C3A\u22efC3A and C7\u2014H7/H7\u2014C7, whereas those in II correspond to C2\u22efC5/C5\u22efC2, C4\u2014H4\u22efC12/ C12\u22efH4\u2014C4, C7A\u22efH6\u2014C6/C6\u2014H6\u22efC7A, C3A\u22efH6\u2014C6/C6-H6\u22efC3A and C3A\u22efC6/C6\u22efC3A contacts. The largest contributions to the Hirshfeld surface of I arises from H\u22efH (44.4%), C\u22efH/H\u22efC (31.9%), and O\u22efH/H\u22efO (13.5%) contacts, whereas the contributions for II are H\u22efH (45.8%), C\u22efH/H\u22efC (27.5%) and O\u22efH/H\u22efO (15.5%). Minor contributions include N\u22efH/H\u22efN (3.6%), C\u22efC (3.2%), C\u22efN/N\u22efC (2.1%), C\u22efO/O\u22efC (1.4%) for I and C\u22efC (5.4%), C\u22efN/N\u22efC (3.4%), N\u22efH/H\u22efN (3.2%), C\u22efO/O\u22efC (0.2%) for II.Minor inter\u00admolecular inter\u00adactions are observed as faint red spots on the surface. The spots in 4.et al., 2016et al., 2013tert-butyl chains. The distances between a nitro\u00adgen donor and an oxygen acceptor range from 2.79\u20132.84\u2005\u00c5, comparable to the values for I and II of 2.7786\u2005(14) and 2.8453\u2005(14)\u2005\u00c5, respectively.A search for the title compound in the Cambridge Structural Database -one was prepared following a reported procedure was stirred at room temperature overnight. The resulting white precipitate was filtered. An additional white precipitate was acquired by adding Et2O (10\u2005mL) into the filtrate. Combined yield: 0.30\u2005g (46%). 1H NMR : \u03b4 10.75 , 7.58 , 7.45 , 7.17 , 7.10 , 7.06 . Pentane vapor diffusion into a solution of the compound in THF formed colorless needles and blocks.3-Phenyl-16.Crystal data, data collection, and refinement statistics are summarized in Table\u00a0310.1107/S2056989023003961/vm2281sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989023003961/vm2281Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989023003961/vm2281IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989023003961/vm2281Isup4.cmlSupporting information file. DOI: 2260424, 2260423CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Reducing the necessary time to restore primary teeth improves the cooperation of paediatric patients. This study aimed to investigate the marginal integrity of restorations prepared with a bulk-fill resin-based composite (RBC) containing additional fragmentation chain transfer (AFCT) compared to a conventional RBC when light cured with a rapid high-irradiance (3\u00a0s) and a regular (10\u00a0s) curing mode.2 or high-irradiance: 3\u00a0s @ 3000 mW/cm2) and the used restorative material . After thermo-mechanical loading, the marginal integrity was analysed using scanning electron microscopy. A beta regression model and pairwise comparisons were used to statistically analyse the data.Forty class-II cavities were prepared in 40 primary molars. The molars were randomly divided into four groups based on the applied light-curing modes (regular: 10\u00a0s @ 1200 mW/cmp\u2009\u2264\u20090.05). The difference between the light-curing modes for the same RBC was not statistically significant (p \u02c3 0.5).The mean marginal integrity (% \u00b1 SD) of the restorations for each group was as follows: Power Fill (10\u00a0s: 79.7\u2009\u00b1\u200915.6) (3\u00a0s: 77.6\u2009\u00b1\u200911.3), Prime (10\u00a0s: 69.7\u2009\u00b1\u200911.1) (3\u00a0s: 75.0\u2009\u00b1\u20099.7). The difference between the RBCs for the same light-curing mode was statistically significant (AFCT-containing bulk-fill RBC \u201cPower Fill\u201d achieves similar marginal integrity when light-cured with either high-irradiance or regular light-curing modes. \u201cPower Fill\u201d achieves better marginal integrity than the conventional RBC \u201cPrime\u201d regardless of the applied light-curing mode. Resin-based composites have become a solid restorative material option in both permanent and primary teeth \u20133. Some 2), and the incorporation of an \u03b2-allyl sulfone addition fragmentation chain transfer (AFCT) reagent in the matrix of the bulk-fill composite [Two major developments led to the possibility of such short polymerisation time: the introduction of high-power light emitting diode (LED) polymerisation units that can produce high radiant exitance and the authorisation from the ethics committee was waived . Roots of the extracted molars were embedded in acrylic resin 3\u00a0mm below the cemento-enamel junction and mounted on custom-made holders. One standardised mesial or distal proximal cavity was prepared in each primary molar. Proximal cavities were prepared with 80-\u00b5m cylindrical burs rotating at 40,000\u00a0rpm in a high-speed contra-angle handpiece . The bur was exchanged after preparing four cavities. The molars were then randomised into four groups based on the RBC they would be restored with and the light-curing mode they would be subjected to. The sample size was determined based on recent similar research [2 . The cavities were restored based on their experimental group as shown in Fig.\u00a02, curing time\u2009=\u200910\u00a0s). In groups 3 and 4, each layer of both materials was subjected to high-irradiance light-curing . The composition of the used RBCs is shown in Table\u00a0The molars were mounted and restored in a custom-made adjacent-tooth simulator with two plastic molars on each side on a laboratory desk. A stainless-steel matrix band was inserted and fixed with a wooden wedge placed 1\u00a0mm below the gingival margin of the cavity. A universal adhesive was scrubbed for 20\u00a0s on all cavity walls in self-etch mode, thinned with a gentle blow of air in order to evaporate the solvent, and then light-cured for 10\u00a0s at 1200 mW/cmAfter the storage time, impressions were taken for each restoration with an A-silicon material . The impressions were poured out , fixed on aluminium holders and the formed replicas were sputter-coated with gold . After sputter coating, each replica was examined under a stereo microscope . The cemento-enamel junction buccally and lingually from the restoration was accentuated inside the epoxy resin using a fine scalpel. The replicas were quantitatively analysed for marginal integrity using scanning electron microscopy (SEM) at 20\u00a0kV and 200 \u00d7 magnification . The marginal integrity for each restoration was expressed as a percentage of continuous margins in relation to the entire length of assessable margins. After this initial marginal analysis, all restorations underwent a thermo-mechanical loading (TML) inside an electronic masticator . The restorations were loaded on their occlusal part using a metal ball (diameter\u2009=\u20093\u00a0mm) for 400 000 loading cycles at 49\u00a0N . SimultaA-silicon impressions were taken again after TML and a final quantitative margin analysis using the same aforementioned protocol was carried out. The margin analysis was conducted by one calibrated and experienced operator (MZ) who was blinded to the groups and had only access to the SEM images. Figure\u00a02 and 3\u00a0s @ 3000 mW/cm2). A beta regression model was set to analyse the data. Post-hoc pairwise comparisons between the groups of the same material with different light-curing modes \u201cPower Fill (10 s)\u201d vs. \u201cPower Fill (3 s)\u201d and \u201cPrime (10 s)\u201d vs. \u201cPrime (3 s)\u201d and between the groups of the same light-curing mode with different materials \u201cPower Fill (10 s)\u201d vs. \u201cPrime (10 s)\u201d and \u201cPower Fill (3 s)\u201d vs. \u201cPrime (3 s)\u201d were computed using the emmeans package [www.R-project.org).The marginal integrity was set as the target variable. This variable was analysed with respect to the restoration type and the light-curing modes (10\u00a0s @ 1200 mW/cm package . The sigp \u02c3 0.1).Before TML, the achieved marginal integrity (% \u00b1 standard deviation SD) of each tested RBC with each tested light-curing mode (regular \u201c10 s\u201d and high-irradiance \u201c3 s\u201d) was as follows: Power Fill (10\u00a0s: 92.1\u2009\u00b1\u200910.2) (3\u00a0s: 90.1\u2009\u00b1\u20095.4), Prime (10\u00a0s: 90.1\u2009\u00b1\u20096.9) (3\u00a0s: 90.3\u2009\u00b1\u20097.3). The difference between \u201cPower Fill (10 s)\u201d vs. \u201cPower Fill (3 s)\u201d and \u201cPrime (10 s)\u201d vs. \u201cPrime (3 s)\u201d and between \u201cPower Fill (10 s)\u201d vs. \u201cPrime (10 s)\u201d and \u201cPower Fill (3 s)\u201d vs. \u201cPrime (3 s)\u201d was not statistically significant (p\u2009\u2264\u20090.05). The difference between \u201cPower Fill (10 s)\u201d vs. \u201cPower Fill (3 s)\u201d, and between \u201cPrime (10 s)\u201d vs. \u201cPrime (3 s)\u201d was not statistically significant (p \u02c3 0.5). Figure\u00a0After TML, the achieved marginal integrity (% \u00b1 SD) of each tested RBC with each tested light-curing mode was as follows: Power Fill (10\u00a0s: 79.7\u2009\u00b1\u200915.6) (3\u00a0s: 77.6\u2009\u00b1\u200911.3), Prime (10\u00a0s: 69.7\u2009\u00b1\u200911.1) (3\u00a0s: 75.0\u2009\u00b1\u20099.7). The difference between \u201cPower Fill (10 s)\u201d vs. \u201cPrime (10 s)\u201d, and between \u201cPower Fill (3 s)\u201d vs. \u201cPrime (3 s)\u201d was statistically significant (p \u02c3 0.1).Before TML, the achieved marginal integrity (% \u00b1 SD) of each tested RBC with each tested light-curing mode was as follows: Power Fill (10\u00a0s: 93.3\u2009\u00b1\u200910.6) (3\u00a0s: 89.5\u2009\u00b1\u20095.3), Prime (10\u00a0s: 89.3\u2009\u00b1\u20098.3) (3\u00a0s: 91.3\u2009\u00b1\u20096.7). The difference between \u201cPower Fill (10 s)\u201d vs. \u201cPower Fill (3 s)\u201d and \u201cPrime (10 s)\u201d vs. \u201cPrime (3 s)\u201d, and between \u201cPower Fill (10 s)\u201d vs. \u201cPrime (10 s)\u201d and \u201cPower Fill (3 s)\u201d vs. \u201cPrime (3 s)\u201d was not statistically significant (p \u02c3 0.1). Figure\u00a0After TML, the achieved marginal integrity (% \u00b1 SD) of each tested RBC with each tested light-curing mode was as follows: Power Fill 10\u00a0s: 83.4\u2009\u00b1\u200916.1) (3\u00a0s: 77.0\u2009\u00b1\u200911.9), Prime (10\u00a0s: 75.8\u2009\u00b1\u200914.1) (3\u00a0s: 76.9\u2009\u00b1\u20099.4). The difference between \u201cPower Fill (10 s)\u201d vs. \u201cPower Fill (3 s)\u201d and \u201cPrime (10 s)\u201d vs. \u201cPrime (3 s)\u201d, and between \u201cPower Fill (10 s)\u201d vs. \u201cPrime (10 s)\u201d and \u201cPower Fill (3 s)\u201d vs. \u201cPrime (3 s)\u201d was not statistically significant (\u2009\u00b1\u200916.1 (p \u02c3 0.5).Before TML, the achieved marginal integrity (% \u00b1 SD) of each tested RBC with each tested light-curing mode was as follows: Power Fill (10\u00a0s: 88.9\u2009\u00b1\u200914.2) (3\u00a0s: 91.3\u2009\u00b1\u20097.0), Prime (10\u00a0s: 92.0\u2009\u00b1\u20094.7) (3\u00a0s: 88.2\u2009\u00b1\u200913.6). The difference between \u201cPower Fill (10 s)\u201d vs. \u201cPower Fill (3 s)\u201d and \u201cPrime (10 s)\u201d vs. \u201cPrime (3 s)\u201d, and between \u201cPower Fill (10 s)\u201d vs. \u201cPrime (10 s)\u201d and \u201cPower Fill (3 s)\u201d vs. \u201cPrime (3 s)\u201d was not statistically significant (p\u2009\u2264\u20090.05). The difference between \u201cPower Fill (10 s)\u201d vs. \u201cPower Fill (3 s)\u201d, and between \u201cPrime (10 s)\u201d vs. \u201cPrime (3 s)\u201d was also statistically significant (p\u2009\u2264\u20090.05). Figure\u00a0After TML, the achieved marginal integrity (% \u00b1 SD) of each tested RBC with each tested light-curing mode was as follows: Power Fill (10\u00a0s: 71.0\u2009\u00b1\u200918.1) (3\u00a0s: 78.8\u2009\u00b1\u200914.0), Prime (10\u00a0s: 57.7\u2009\u00b1\u200915.4) (3\u00a0s: 71.2\u2009\u00b1\u200915.0). The difference between \u201cPower Fill (10 s)\u201d vs. \u201cPrime (10 s)\u201d, and between \u201cPower Fill (3 s)\u201d vs. \u201cPrime (3 s)\u201d was statistically significant (2) on the marginal integrity of an AFCT-containing bulk-fill RBC and a conventional RBC in standardised deep class-II cavities in primary molars. The marginal integrity in this study was not affected by the different light-curing modes (the first null-hypothesis cannot be rejected), but indeed affected by the tested RBC (the second null-hypothesis must be rejected).This in-vitro study investigated the effect of a high-irradiance light curing in the present study. The first attempts to evaluate dental restorative materials with regard to their marginal integrity date back to the 1970s . Some stThe here used loading force and frequency is reported to be most frequently used to simulate the mechanical degradation of resin composite restorations in vitro . HoweverTaking the margins of the whole restoration (enamel and dentine) into consideration, employing the high-irradiance light-curing mode resulted in similar marginal integrities as the regular 10-s light-curing mode, regardless of the tested RBC. As for Power Fill, this result could be attributed to the presence of the AFCT reagent, which allows a step-like progression of the polymer chain and enhances the thermal and mechanical properties of RBCs, and to the fact that this RBC was actually developed to be compatible with high-irradiance light curing . Par et The bulk-fill RBC \u201cPower Fill\u201d showed better marginal integrity than the conventional RBC \u201cPrime\u201d regardless of the used light-curing mode. This is in contrast to the results of a recent study, which compared the same RBCs and concluded that both achieve similar marginal integrity in primary molars . This co2) does not actually help explain the observed similar marginal integrity. Nevertheless, Steffen et al. [2) or high-irradiance (3\u00a0s @ 2850 mW/cm2) modes. Similar findings (regarding shrinkage stress and degree of conversion) for AFCT-free RBCs were also reported by Par et al. [The conventional RBC \u201cPrime\u201d was not especially developed for high-irradiance light curing and does not contain an AFCT reagent. This makes the observed similar marginal integrity between both applied light-curing modes rather interesting. The fact that the radiant exposure in case of the high-irradiance light-curing mode is less than the regular light-curing mode or regular light-curing modes (10\u00a0s @ 1200 mW/cm2).AFCT-containing high-viscous bulk-fill RBC \u201cPower Fill\u201d achieves similar performance with regard to marginal integrity of deep class-II cavities in primary molars when light-cured with high-irradiance (3\u00a0s @ 3000 mW/cmAFCT-containing high-viscous bulk-fill RBC \u201cPower Fill\u201d achieves better performance than the conventional RBC \u201cPrime\u201d with regard to marginal integrity in deep class-II cavities in primary molars regardless of the used light-curing mode."} +{"text": "Plasmodium falciparum, is driven by specific transcriptional programs, but it is unclear how most genes are activated or silenced at specific times. There is an association between transcription and spatial organization; however, the molecular mechanisms behind genome organization are unclear. While P.\u2009falciparum lacks key genome\u2010organizing proteins found in metazoans, it has all core components of the cohesin complex. To investigate the role of cohesin in P.\u2009falciparum, we functionally characterize the cohesin subunit Structural Maintenance of Chromosomes protein 3 (SMC3). SMC3 knockdown during early stages of the intraerythrocytic developmental cycle (IDC) upregulates a subset of genes involved in erythrocyte egress and invasion, which are normally expressed at later stages. ChIP\u2010seq analyses reveal that during the IDC, SMC3 enrichment at the promoter regions of these genes inversely correlates with gene expression and chromatin accessibility. These data suggest that SMC3 binding contributes to the repression of specific genes until their appropriate time of expression, revealing a new mode of stage\u2010specific gene repression in P.\u2009falciparum.The complex life cycle of the human malaria parasite, Plasmodium falciparum, the cohesin subunit SMC3 plays a role in stage\u2010specific gene repression that is distinct from heterochromatin\u2010mediated repression of genes involved in antigenic variation and gametocytogenesis.In the human malaria parasite Plasmodium falciparum, has a significant impact on human health in endemic regions was found to play a role in the nuclear organization of centromeres, and knockdown led to defects in var gene transcription \u2014key genome organizing proteins in metazoans , SMC3 (PF3D7_0414000), and a protein with the N\u2010terminal Rad21/Rec8 domain is present in the P. falciparum cohesin complex and stag , 24 (trophozoite), and 36 (schizont) hours post invasion (hpi). Using the macs2 peak calling algorithm between significant peaks called in both clones for each time point, and we used this cohort of shared peaks for further analysis of SMC3\u20103HA\u2010 Dataset\u00a0. No signet\u00a0al,\u00a0et\u00a0al\u00a0, \u201ccell\u2013cell adhesion\u201d , \u201cresponse to host\u201d , and \u201cantigenic variation\u201d , \u201cmovement in host environment\u201d (q\u2009=\u20093.3\u2009\u00d7\u200910\u22123), \u201cexit from host\u201d (q\u2009=\u20090.026), and \u201centry into host\u201d (q\u2009=\u20090.05). These categories include genes that are involved in invasion of or egress from the red blood cell such as ralp1 , as knockdown at the protein level could not be achieved after a single cell cycle , 69% at 24 hpi (Dataset\u00a0q\u2009=\u20091.3\u2009\u00d7\u200910\u221239), and 48% at 36 hpi and ribonucleoprotein complex biogenesis (1.82\u2009\u00d7\u200910\u221215), subunit organization (2.47\u2009\u00d7\u200910\u221213), and assembly , glideosome\u2010associated protein 45 , merozoite surface protein 1 , and merozoite surface protein 9 does not inhibit parasite growth agrees with reports in S. cerevisiae and D. melanogaster in which normal growth and sister chromatid cohesion were achieved despite an 87 and 80% decrease, respectively, in Rad21, a core component of the cohesin complex datasets do not provide evidence of typical enhancer\u2010promoter interactions found in other eukaryotes is involved in transcription of invasion\u2010related genes via binding to the GTGCA motif, likely by interaction with the bromodomain protein 1 lysis during ring stage followed by a plasmagel enrichment for late blood stages 24\u2009h later. Another sorbitol treatment 6\u2009h afterward places the 0\u2009h time point 3\u2009h after the plasmagel enrichment. Thus, the window of synchronicity for cultures is \u00b1\u20093\u2009h. Parasite development was monitored by Giemsa staining. Parasites were harvested at 1\u20135% parasitemia.Blood stage 3D7 io et\u00a0al\u00a0. Brieflyglms strain was generated using a two\u2010plasmid system (pUF1 and pL7) based on the CRISPR/Cas9 system previously described in Ghorbal et\u00a0al\u00a0(PfSMC3\u20103HA\u2010glmS containing the single guide RNA (sgRNA) \u2010encoding sequence 5\u2032\u2010CCTAGAAAATTAGAACAATT\u20103\u2032 targeting the 3\u2032 UTR of PF3D7_0414000. The pL7\u2010PfSMC3\u20103HA\u2010glmS plasmid also contained the homology repair construct 5\u2032\u2010AGATAGAGAGAGTTATATATCTAAAGGAACAAAGAATGAGGCCTACGAAATTATTAGCATTGTATAAAAAAAAAAAGAAAAAAAAAAGAAAAAAAAAAAAGATTATATATATAATATATGTTGACAATTAATAAATATATTTGTATATATCTGTTAACTAATTATGAAAATTTTTGAATCAATAAATTTTTTAAATAACAAAAAAAAAAAAAAATATATATATTATATATATATTTTATATTTTATATTTTCTTGTAATTTTTGTTTTTTTAGGAGGAAAAACATGCCCTAGAAAATggcggtggaTACCCTTACGATGTGCCTGATTACGCGTAtCCcTAtGAcGTaCCaGAcTAtGCGTACCCtTAtGAcGTtCCgGATTAtGCtcacggggtgTAAGCGGCCGCGGTCTTGTTCTTATTTTCTCAATAGGAAAAGAAGACGGGATTATTGCTTTACCTATAATTATAGCGCCCGAACTAAGCGCCCGGAAAAAGGCTTAGTTGACGAGGATGGAGGTTATCGAATTTTCGGCGGATGCCTCCCGGCTGAGTGTGCAGATCACAGCCGTAAGGATTTCTTCAAACCAAGGGGGTGACTCCTTGAACAAAGAGAAATCACATGATCTTCCAAAAAACATGTAGGAGGGGACAACAATTTGGTTTTGTTTTTTTCTTTAGGTTTTGAGAAAAACAAATAGGAAATACAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAATGTATTTTTACATATGCACTTGGATTATTTTATTTTTATTATTTTTCTTTATATAAAGTAAAAATATACATAAGTATGCTTATTTATTACATAAGAGTTTATTTAAGAAAGGTTTCTTTTTCATAATATTGTGTGCATGAGTTTTTTTTTATTTTATTTTTTTTTTTTATTTCTGTAACGAAAAGGATATTAAAAAAAATAATAAAA\u20103\u2032 . This homology repair construct comprises a 3\u2009\u00d7\u2009Hemaglutinin (3HA) \u2013 encoding sequence followed by a glmS ribozyme\u2010encoding sequence and 1.5\u2009\u03bcM DSM1 (MR4/BEI Resources). Parasites reappeared approximately 3\u2009weeks after transfection, and 5\u2010fluorocytosine was used to negatively select the pL7 plasmid.The SMC3\u20103HA\u2010al et\u00a0al\u00a0. A 3D7 wet\u00a0al\u00a0(smc1 (PF3D7_1130700) and stag (PF3D7_1456500), respectively, were amplified using either SMC1\u2010HR\u2010FW/RV or STAG\u2010HR\u2010FW/RV (Dataset\u00a0The SMC1\u20103HA\u2010dd and STAG\u20103HA\u2010dd strains were generated\u00a0using the method of selection\u2010linked integration (SLI) previously\u00a0described in Birnbaum et\u00a0al\u00a0. Homolog Dataset\u00a0. The obt Dataset\u00a0, followeE. coli (Agilent Technologies 200315). All the parasite lines were cloned by limiting dilution, and integration at the targeted genomic locus was confirmed by PCR (Dataset\u00a0All cloning was performed using KAPA HiFi DNA Polymerase (Roche 07958846001), In\u2010Fusion HD Cloning Kit (Clontech 639649), and XL10\u2010Gold Ultracompetent Dataset\u00a0 and Sangglms clone and WT culture , as a negative control, were synchronized. Late stage parasites (1.5\u2009\u00d7\u2009109 parasites) were enriched using Percoll density gradient separation and then cross\u2010linked with 1\u2009ml 0.5\u2009mM dithiobissuccinimidyl propionate in DPBS for 60\u2009min at 37\u00b0C containing protease and phosphatase inhibitor cocktail (Thermo Fisher 78440) and 1\u2009U/\u03bcl of Benzonase (Merck 71206). The lysates were cleared by centrifugation at 16,000\u2009g for 10\u2009min at 4\u00b0C. Supernatants were incubated with 25\u2009\u03bcl of anti\u2010HA Dynabeads (Thermo Fisher 88836) overnight with rotation at 4\u00b0C. Beads were collected with a magnet and washed five times with 1\u2009ml RIPA buffer, then five times with 1\u2009ml DPBS, and then once with 1\u2009ml 25\u2009mM NH4HCO3 (Sigma 09830). The beads were reduced with 100\u2009mM dithiothreitol (Sigma D9779), alkylated with 55\u2009mM iodoacetamide (Sigma I1149), and subjected to on\u2010bead digestion using 1\u2009\u03bcg of trypsin (Thermo Fisher 90059). The resulting peptides were desalted using C18 ziptips (Merck ZTC04S096) and sent for MS analysis.An SMC3\u20103HA\u2010m/z, AGC 1e5 using an EASY\u2010Spray column, 50\u2009cm\u2009\u00d7\u200975\u2009\u03bcm ID, PepMap RSLC C18, 2\u2009\u03bcm (Thermo Fisher ES803A) over a 70\u2010min gradient before nanoelectrospray using a Q Exactive HF\u2010X mass spectrometer (Thermo Fisher). The mass spectrometer was operated in a data\u2010dependent mode. The parameters for the full scan MS were as follows: resolution of 60,000 across 350\u20131,500\u20092, 1% IGEPAL CA\u2010630, and 1\u00d7 protease inhibitor cocktail ) at 4\u00b0C and incubated on ice for 30\u2009min. The cytoplasmic lysate was cleared with centrifugation . The pellet (containing the nuclei) was resuspended in 3.3 times less volume of nuclear extraction buffer than cytoplasmic lysis buffer at 4\u00b0C, transferred to 1.5\u2009ml sonication tubes , and sonicated for 5\u2009min total (10\u2009cycles of 30\u2009s on/off) in a Diagenode Pico Bioruptor at 4\u00b0C. This nuclear lysate was cleared with centrifugation . Protein samples were supplemented with NuPage Sample Buffer (Thermo Fisher NP0008) and NuPage Reducing Agent (Thermo Fisher NP0004) and denatured for 10\u2009min at 70\u00b0C. Proteins were separated on a 4\u201312% Bis\u2010Tris NuPage gel (Thermo Fisher NP0321) and transferred to a PVDF membrane with a Trans\u2010Blot Turbo Transfer system (Bio\u2010Rad). The membrane was blocked for 1\u2009h with 1% milk in PBST at 25\u00b0C. HA\u2010tagged proteins and histone H3 were detected with anti\u2010HA and anti\u2010H3 primary antibodies, respectively, followed by donkey anti\u2010rabbit secondary antibody conjugated to horseradish peroxidase . SMC3 was detected with an in\u2010house\u2010generated anti\u2010SMC3 antibody , followed by sheep HRP anti\u2010mouse secondary antibody .\u00a0HRP signal was developed with SuperSignal West Pico Plus or Femto chemiluminescent substrate and imaged with a ChemiDoc XRS+ (Bio\u2010Rad).Parasites were washed once with Dulbecco's phosphate\u2010buffered saline , then resuspended in cytoplasmic lysis buffer was amplified from cDNA of 3D7 wild\u2010type parasites and inserted into the pGEX expression vector. The plasmid was verified by sequencing and used to transform SHuffle T7 expression bacteria (New England BioLabs C3026J). Recombinant protein expression was induced with 100\u2009\u03bcM IPTG overnight with shaking at 16\u00b0C, and the pellet was frozen at \u221280\u00b0C until use. Soluble GST\u2010tagged recombinant protein was purified using Glutathione Sepharose 4 Fast Flow (Cytiva GE17\u20105132\u201003), according to manufacturer's instructions. Briefly, the bacterial pellet was resuspended in cold lysis buffer . The lysate was sonicated on ice with a probe sonicator . The sample was centrifuged at 18,000\u2009g for 1\u2009h at 4\u00b0C, and the soluble lysate was incubated with Glutathione Sepharose beads (Cytiva GE17\u20105132\u201003) overnight with gentle tumbling at 4\u00b0C. The next day, the lysate with beads was loaded onto chromatography columns , and the beads were washed with cold PBS containing 5\u2009mM EDTA. Recombinant protein was eluted with elution buffer , buffer exchanged into PBS , and stored at \u221220\u00b0C until use. Final protein concentration was obtained using Protein Assay (BioRad).The coding sequence for the N\u2010terminal region (amino acids 1\u2010253) of th, 2013). All experiments were approved by the Ethics Committee CETEA Institute Pasteur and registered under the reference Permit Number dap180040, issued in 2018. The PfSMC3 antibody was purified from pooled mouse serum using rProtein G Agarose (Invitrogen 15920) according to manufacturer's instructions. The final purified antibody was dialyzed in PBS using a Float\u2010A\u2010Lyzer (Repligen). Purified antibodies were stored at \u221220\u00b0C in 50% Glycerol.Purified recombinant protein was used to immunize C57BL/6 mice. Ten micrograms were used for the first immunization in Freund's Complete Adjuvant (Sigma F5881) and three subsequent boosters in Freund's Incomplete Adjuvant (Sigma F5506). Animals were housed in the Institute Pasteur animal facilities, accredited by the French Ministry of Agriculture for performing experiments on live rodents. Work on animals was performed in compliance with French and European regulations on care and protection of laboratory animals . Parasite cultures were centrifuged at 800\u2009g for 3\u2009min at 25\u00b0C. Medium was removed and the RBCs were lysed with 10\u2009ml 0.075% saponin (Sigma S7900) in DPBS at 37\u00b0C. The parasites were centrifuged at 3,250\u2009g for 3\u2009min at 25\u00b0C and washed twice with 10\u2009ml DPBS at 4\u00b0C. The supernatant was removed, and the parasite pellet was resuspended in 1\u2009ml 0.5\u2009mM dithiobissuccinimidyl propionate in DPBS for 30\u2009min at 25\u00b0C were washed twice with ChIP dilution buffer using a DynaMag magnet (Thermo Fisher 12321D). The beads were resuspended in 200\u2009\u03bcl 0.02% Tween\u201020 in DPBS with 1\u2009\u03bcg of anti\u2010HA antibody (Abcam ab9110) or IgG (Diagenode C15410206) and incubated with rotation at 4\u00b0C for 6\u2009h.g and the supernatant was removed. The pellet was resuspended in 1\u2009ml cytoplasmic wash buffer and rotated for 10\u2009min. The lysates were centrifuged for 5\u2009min at 5,000\u2009g, and the supernatant was removed. The pellet was resuspended in 200\u2009\u03bcl nuclear lysis buffer , transferred to 1.5\u2009ml sonication tubes (Diagenode C30010016), and sonicated for 2.5\u2009min total (5\u2009cycles of 30\u2009s on/off) in a Diagenode Pico Bioruptor. The sonicated extracts were centrifuged at 13,500\u2009g for 10\u2009min. The supernatant was diluted with 400\u2009\u03bcl dilution buffer 2 .All further steps were performed at 4\u00b0C. The cross\u2010linked parasites were resuspended in 900\u2009\u03bcl of cytoplasmic lysis buffer and rotated for 30\u2009min. The lysates were centrifuged for 5\u2009min at 2,000\u2009PfSMC3 antibody (1:500 in 1% milk\u2010PBST) followed by sheep HRP anti\u2010mouse secondary antibody . Histone H3 was detected with Anti\u2010H3 , followed by donkey HRP anti\u2010rabbit secondary antibody . HRP signal was developed with SuperSignal West Pico Plus or West Femto chemiluminescent substrate and imaged with a ChemiDoc XRS+ (Bio\u2010Rad).Using a DynaMag magnet, the antibody\u2010conjugated Dynabeads were washed once with 200\u2009\u03bcl of dilution buffer 1 and resuspended in 200\u2009\u03bcl dilution buffer 2 . The washed antibody\u2010conjugated Dynabeads were added to the diluted nuclear lysate and incubated overnight with rotation at 4\u00b0C. The beads were collected on a DynaMag magnet and the supernatant was removed and kept as flow through. The beads were washed three times with 200\u2009\u03bcl of wash buffer . The beads were resuspended in 30\u2009\u03bcl of 2\u00d7 NuPage Sample Buffer (Thermo Fisher NP0008) and 3.4\u2009\u03bcl of NuPage Reducing Agent (Thermo Fisher NP0004) and denatured for 5\u2009min at 95\u00b0C. The eluate was separated from the beads with the DynaMag magnet. Lysates and the bead eluate were separated on a 3\u20108% Tris\u2010Acetate NuPage gel (Thermo Fisher EA0375) and transferred to a PVDF membrane with a Trans\u2010Blot Turbo Transfer system (Bio\u2010Rad). Membranes were blocked with 1% milk\u2010PBST . HA\u2010tagged SMC1 and STAG proteins were detected with anti\u2010HA conjugated to HRP . SMC3 was detected with anti\u2010et\u00a0al,\u00a0et\u00a0al\u00a0 in PBS for 30\u2009min. Cells were incubated with anti\u2010HA and/or anti\u2010HP1 [from with DAPI for 45\u2009min followed by three 10\u2009min washes with 0.5% Tween\u00ae20/PBS. Cells were washed once more with PBS and placed onto polyethyleneimine\u2010coated slides (Thermo Scientific 30\u201042H\u2010RED\u2010CE24). Once adhered to the slide, cells were washed twice with PBS and mounted with VectaShield\u00ae (Vector Laboratories). Images were acquired using a Deltavision Elite imaging system , and Fiji (http://fiji.sc) was used for analysis using the least manipulation possible.iRBCs were washed once with DPBS (Thermo Fisher 14190) at 37\u00b0C and fixed in suspension in 4% paraformaldehyde (EMS 15714) with 0.0075% glutaraldehyde (EMS 16220) in PBS for 20\u2009min at 25\u00b0C, as described previously , 24 and 36 hpi . Parasite culture was centrifuged at 800\u2009g for 3\u2009min at 25\u00b0C. Medium was removed and the RBCs were lysed with 10\u2009ml 0.075% saponin (Sigma S7900) in DPBS at 37\u00b0C. The parasites were centrifuged at 3,250\u2009g for 3\u2009min at 25\u00b0C and washed with 10\u2009ml DPBS at 37\u00b0C. The supernatant was removed, and the parasite pellet was resuspended in 10\u2009ml of PBS at 25\u00b0C. The parasites were cross\u2010linked by adding methanol\u2010free formaldehyde and incubating with gentle agitation for 10\u2009min at 25\u00b0C. The cross\u2010linking reaction was quenched by adding glycine and incubating with gentle agitation for 5\u2009min at 25\u00b0C. Parasites were centrifuged at 3,250\u2009g for 5\u2009min at 4\u00b0C and the supernatant removed. The pellet was washed with DPBS and centrifuged at 3,250\u2009g for 5\u2009min at 4\u00b0C. The supernatant was removed, and the cross\u2010linked parasite pellets were snap\u2010frozen.Two clonal populations (clone A and B) of SMC3\u20103HA\u2010For each time\u2010point, 200\u2009\u03bcl of Protein G Dynabeads (Invitrogen 10004D) were washed twice with 1\u2009ml ChIP dilution buffer using a DynaMag magnet (Thermo Fisher 12321D). The beads were resuspended in 1\u2009ml ChIP dilution buffer with 8\u2009\u03bcg of anti\u2010HA antibody (Abcam ab9110) and incubated on a rotator at 4\u00b0C for 6\u2009h.g at 4\u00b0C, the supernatant was removed, and the pellet was resuspended in SDS lysis buffer at 4\u00b0C (3.6\u2009ml for the 12 hpi sample and 1.8\u2009ml for the 24 ad 36 hpi samples). The liquid was distributed into 1.5\u2009ml sonication tubes and sonicated for 12\u2009min total (24\u2009cycles of 30\u2009s on/off) in a Diagenode Pico Bioruptor at 4\u00b0C. The sonicated extracts were centrifuged at 13,500\u2009g for 10\u2009min at 4\u00b0C and the supernatant, corresponding to the chromatin fraction, was kept. The DNA concentration for each time point was determined using the Qubit dsDNA High Sensitivity Assay Kit (Thermo Fisher Scientific Q32851) with a Qubit 3.0 Fluorometer (Thermo Fisher Scientific). For each time point, chromatin lysate corresponding to 100\u2009ng of DNA was diluted in SDS lysis buffer and kept as \u201cinput\u201d at \u221220\u00b0C. For clones A and B, chromatin lysate corresponding to 19 and 18\u2009\u03bcg (12 hpi), 2 and 1.4\u2009\u03bcg (24 hpi) and 3 and 5.2\u2009\u03bcg (36 hpi) of DNA, respectively,\u00a0was diluted 1:10 in ChIP dilution buffer at 4\u00b0C.The cross\u2010linked parasites were resuspended in 4\u2009ml of lysis buffer at 4\u00b0C, and 10% Nonidet\u2010P40 was added . The parasites were lysed in a prechilled dounce homogenizer (200 strokes for 12 hpi parasites and 100 strokes for 24 and 36 hpi parasites). The lysates were centrifuged for 10\u2009min at 13,500\u2009Low salt wash buffer at 4\u00b0C.High salt wash buffer at 4\u00b0C.LiCl wash buffer at 4\u00b0C.TE wash buffer at 25\u00b0C.Using a DynaMag magnet, the antibody\u2010conjugated Dynabeads were washed twice with 1\u2009ml ChIP dilution buffer and resuspend in 100\u2009\u03bcl of ChIP dilution buffer at 4\u00b0C. Then the washed antibody\u2010conjugated Dynabeads were added to the diluted chromatin sample and incubated overnight with rotation at 4\u00b0C. The beads were collected on a DynaMag into eight different tubes per sample, the supernatant was removed, and the beads in each tube were washed for 5\u2009min with gentle rotation with 1\u2009ml of the following buffers, sequentially:After the washes, the beads were collected on a DynaMag, the supernatant was removed, and the beads for each time point were resuspended in 800\u2009\u03bcl of elution buffer and incubated at 65\u00b0C for 30\u2009min with agitation . The beads were collected on a DynaMag and the eluate, corresponding to the \u201cChIP\u201d samples, was transferred to a different tube.g at 4\u00b0C to separate phases. The aqueous top layer was transferred to another tube and mixed with 30\u2009\u03bcg glycogen (Thermo Fisher 10814) and 5\u2009M NaCl . Eight hundred microliters 100% EtOH at 4\u00b0C were added to each sample, which was then incubated at \u221220\u00b0C for 30\u2009min. The DNA was pelleted by centrifugation for 10\u2009min at 13,500\u2009g at 4\u00b0C, washed with 500\u2009\u03bcl 80% EtOH at 4\u00b0C, and centrifuged for 5\u2009min at 13,500\u2009g at 4\u00b0C. After removing the EtOH, the pellet was dried at 25\u00b0C and all DNA for each sample was resuspended in 30\u2009\u03bcl 10\u2009mM Tris\u2013HCl, pH 8 total. The DNA concentration and average size of the sonicated fragments was determined using a DNA high sensitivity kit (Agilent 5067\u20104626) and the Agilent 2100 Bioanalyzer. Libraries for Illumina Next Generation Sequencing were prepared with the MicroPlex library preparation kit (Diagenode C05010014), with KAPA HiFi polymerase (KAPA biosystems) substituted for the PCR amplification. Libraries were sequenced on the NextSeq 500 platform (Illumina).For purification of the DNA, both \u201cChIP\u201d and \u201cInput\u201d samples were incubated for approximately 10\u2009h at 65\u00b0C to reverse the crosslinking. Two hundred microliters of TE buffer followed by 8\u2009\u03bcl of RNaseA were added to each sample, which was then incubated for 2\u2009h at 37\u00b0C. Four microliters Proteinase K were added to each sample, which was then incubated for 2\u2009h at 55\u00b0C. Four hundered microliters phenol:chloroform:isoamyl alcohol were added to each sample, which was then mixed with vortexing and centrifuged for 10\u2009min at 13,500\u2009P.\u00a0falciparum genome were mapped to the P. falciparum reference genome feature file was added to one half of the culture for two rounds of parasite replication (approximately 96\u2009h). Parasites were then re\u2010synchronized and three technical replicates (with and without glucosamine) were harvested at 12, 24, and 36 hpi. RBCs were lysed in 0.075% saponin (Sigma S7900) in PBS at 37\u00b0C, centrifuged at 3,250\u2009g for 5\u2009min, washed in PBS, centrifuged at 3,250\u2009g for 5\u2009min, and resuspended in 700\u2009\u03bcl QIAzol reagent (Qiagen 79306). RNA was extracted using an miRNeasy Mini kit (Qiagen 1038703) with the recommended on\u2010column DNase treatment. Total RNA was poly (A) selected using the Dynabeads mRNA Purification Kit (Thermo Fischer Scientific 61006). Library preparation was performed with the NEBNext\u00ae Ultra\u2122 II Directional RNA Library Prep Kit for Illumina\u00ae (New England Biolabs E7760S) and paired end sequencing was performed on the Nextseq 550 platform (Illumina). Sequenced reads were mapped to the P. falciparum genome using the built\u2010in tool at PlasmoDB.org .A WT and SMC3\u20103HA\u2010et\u00a0al,\u00a0et\u00a0al,\u00a0et\u00a0al\u00a0 of each sample to the microarray data from was added to one half for approximately 96\u2009h before starting the growth curve. The parasites were tightly re\u2010synchronized and diluted to ~0.2% parasitemia (5% hematocrit) at ring stage. The growth curve was performed in a 96\u2010well plate (200\u2009\u03bcl culture per well) with three technical replicates per condition per blood. Every 24\u2009h, 5\u2009\u03bcl of the culture were fixed in 45\u2009\u03bcl of 0.025% glutaraldehyde in PBS for 1\u2009h at 4\u00b0C. After centrifuging at 800\u2009g for 5\u2009min, free aldehyde groups were quenched by re\u2010suspending the iRBC pellet in 200\u2009\u03bcl of 15\u2009mM NH4Cl in PBS. A 1:10 dilution of the quenched iRBC suspension was incubated with Sybr Green I (Sigma S9430) to stain the parasite nuclei. Quantification of the iRBCs was performed in a CytoFLEX S cytometer (Beckman Coulter), and analysis with was done with FlowJo Software. Growth rate was calculated and expressed as the parasitemia at Day X divided by the starting average parasitemia (Day 1) of the correspondent culture.Parasite growth was measured as described previously (Vembar BioRender.com.The synopsis image for this manuscript was created with Catarina Rosa: Conceptualization; investigation; visualization; writing \u2013 original draft; writing \u2013 review and editing. Parul Singh: Conceptualization; data curation; formal analysis; validation; visualization; writing \u2013 original draft; writing \u2013 review and editing. Patty Chen: Investigation; visualization. Ameya Sinha: Investigation. Aur\u00e9lie Cla\u00ebs: Investigation. Peter R Preiser: Supervision; funding acquisition. Peter C Dedon: Supervision; funding acquisition. Sebastian Baumgarten: Conceptualization; formal analysis; supervision; validation; visualization; writing \u2013 original draft; writing \u2013 review and editing. Artur Scherf: Supervision; funding acquisition. Jessica M Bryant: Conceptualization; formal analysis; supervision; funding acquisition; visualization; writing \u2013 original draft; project administration; writing \u2013 review and editing.The authors declare that they have no conflict of interest.Expanded View Figures PDFClick here for additional data file.Dataset EV1Click here for additional data file.Dataset EV2Click here for additional data file.Dataset EV3Click here for additional data file.Dataset EV4Click here for additional data file.Dataset EV5Click here for additional data file.Dataset EV6Click here for additional data file.Dataset EV7Click here for additional data file.Dataset EV8Click here for additional data file.Dataset EV9Click here for additional data file.Dataset EV10Click here for additional data file.Dataset EV11Click here for additional data file.Dataset EV12Click here for additional data file.Dataset EV13Click here for additional data file.Dataset EV14Click here for additional data file.Dataset EV15Click here for additional data file.Dataset EV16Click here for additional data file.Dataset EV17Click here for additional data file.Dataset EV18Click here for additional data file.Dataset EV19Click here for additional data file.Dataset EV20Click here for additional data file.Dataset EV21Click here for additional data file.Dataset EV22Click here for additional data file.PDF+Click here for additional data file.Source Data for Figure 1Click here for additional data file.Source Data for Figure 2Click here for additional data file.Source Data for Figure 4Click here for additional data file."} +{"text": "The crystal structures of a di\u00adhydro\u00adfurylsilane and a di\u00adhydro\u00adfurylgermane are reported. Hirshfeld surface analyses were performed to investigate the inter\u00admolecular inter\u00adactions. 4H5O)4 (1) and Ge(C4H5O)4 (2) are di\u00adhydro\u00adfuryl compounds of silicon and germanium and are useful building blocks for the functionalization of these elements. Both structures crystallize in space group P21/n in the monoclinic crystal system with two mol\u00adecules in the asymmetric unit: the Si and Ge atoms adopt slightly distorted tetra\u00adhedral geometries, while the C4H5O moieties exhibit shallow envelope conformations. Through a Hirshfeld surface analysis of the structures, inter\u00adactions within the crystal packing could be elucidated: compound 1 features a polymeric chain in the (101) plane via C\u2014H\u22efO hydrogen bonds whereas in 2 C\u2014H\u22efO hydrogen bonds create a polymeric chain in the (010) plane.The title compounds Si(C The two compounds are already known in the literature were prepared by Lukevits and co-workers in the 1980s \u2212x, \u2212y, \u2212z] with a distance of 2.22\u2005(4)\u2005\u00c5 was identified by the red spots on the Hirshfeld surface. However, these red spots show only a small proportion of the inter\u00adactions indicated in the fingerprint. Furthermore, a C30\u22efH24Bii van der Waals inter\u00adaction with a separation of 2.796\u2005(16)\u2005\u00c5 was also identified. The contribution of the C\u22efH inter\u00adactions is 10.7%, which is a low contribution to the crystal packing. Besides these inter\u00adactions, H\u22efO inter\u00adactions could be identified and contribute 19.2% of the structure in the solid state. Hydrogen bonds between C\u2014H\u22efO, which are indicated by red spots on the Hirshfeld surface are listed in Table\u00a05A\u22efO5ii, C27\u2014H27B\u22efO3iii and C31\u2014H31A\u22efO4 can be described as having a D11(2) graph-set motif. C23\u2014H23B\u22efO5i is described by et al., 1990B\u22efO3iii and C31\u2014H31A\u22efO4, a part of the crystal packing is defined along the [101] direction \u2005\u00c5 and H7B\u22efH7Bi at 2.20\u2005(4)\u2005\u00c5 are visible as red spots and could be identified as close inter\u00adactions by the Hirshfeld surface. The contribution of the van der Waals inter\u00adactions is slightly lower at 10.0%. The inter\u00adaction C32\u22efH4Aii at 2.79\u2005(2)\u2005\u00c5 could also be detected on the Hirshfeld surface. As in the case of 1, inter\u00adactions of the form H\u22efO could be determined, which contribute 18.3% to the crystal packing. These hydrogen bonds were detected by red spots on the Hirshfeld surface and are shown in Table\u00a06A\u22efO7i and C23\u2013H23A\u22efO4ii can be described by the graph-set motif D11(2). In contrast, the hydrogen bond C31\u2014H31A\u22efO7iii is described by the graph-set motif et al., 1990A\u22efO7iii and C31\u2014H31A\u22efO7i, a part of the crystal packing, which forms a plane in the [010] direction, can be seen in Fig.\u00a08For the Hirshfeld surface analysis of ed Fig.\u00a06. The dised Fig.\u00a06. The dis4.et al., 2016et al., 2022et al., 20201 and 2. They also display similar (DHF)C\u2014Si\u2014C(DHF) bond angles and also a slightly distorted tetra\u00adhedron. In addition, a deviation in the planarity of the di\u00adhydro\u00adfuryl rings was found there. An extended search for 3-silanes revealed the compounds (trimeth\u00adyl)silane -5-methyl-4-(t-butyl\u00addiphenyl\u00adsil\u00adyl)-2,3-di\u00adhydro-furan-2-carb\u00adoxy\u00adlic acid (1\u2032-phenyl\u00adeth\u00adyl)amide -furan\u00adone silane was also found in the database when searching for (2-furan\u00adyl)silane germane and an extended search for 3-germane found no hits.5.1 and 2 have already been described by Lukevits and Ertschak , tert-butyl\u00adlithium was added at 228\u2005K to a solution of 2,3-di\u00adhydro\u00adfuran in diethyl ether (approx. 100\u2005ml). The reaction solution was stirred for 1\u2005h at room temperature. Then, tetra\u00adchloro\u00adsilane was added at 243\u2005K and the reaction solution was stirred for 1\u2005h. The resulting solid was separated by inert filtration. The obtained solution was concentrated in vacuo and crystallized at 243\u2005K. The solvent was removed and the solid was washed with cold diethyl ether. The product tetra\u00adkis\u00adsilane (1) was obtained as colourless blocks.Compound 1H NMR: : \u03b4 = 2.25 , 4.06 , 5.88 ppm. {1H}13C NMR: : \u03b4 = 31.4 , 71.0 , 117.8 , 155.1 ppm. {1H}29Si NMR: : \u03b4 = \u221251.40 ppm.2), tert-butyl\u00adlithium was added at 228\u2005K to a solution of 2,3-di\u00adhydro\u00adfuran in diethyl ether (approx. 100\u2005ml). The reaction solution was stirred for 1\u2005h at rt. Tetra\u00adchloro\u00adgermane was added at 213\u2005K and the reaction solution was stirred for 1\u2005h. The resulting solid was separated by inert filtration. The obtained solution was concentrated in vacuo and crystallized at 243\u2005K. The solvent was removed, and the solid was washed with cold diethyl ether. The product tetra\u00adkis\u00adgermane (2) was obtained as colourless blocks.For the synthesis of tetra\u00adkis\u00adgermane : \u03b4 = 2.26 , 4.05 , 5.62 ppm. {1H}13C NMR: : \u03b4 = 30.8 , 71.0 , 113.8 , 155.7 ppm.6.A,B, H23A,B, H24A,B, H27A,B and H31A,B for compound 1 and all H atoms for compound 2 were refined independently. Other H atoms were positioned geometrically (C\u2014H = 0.95\u20131.00\u2005\u00c5) and were refined using a riding model, with Uiso(H) = 1.2Ueq(C) for CH2 and CH hydrogen.Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S2056989023003158/hb8058sup1.cifCrystal structure: contains datablock(s) 1. DOI: 10.1107/S2056989023003158/hb80581sup2.hklStructure factors: contains datablock(s) 1. DOI: Click here for additional data file.10.1107/S2056989023003158/hb80581sup4.cmlSupporting information file. DOI: 10.1107/S2056989023003158/hb80582sup3.hklStructure factors: contains datablock(s) 2. DOI: Click here for additional data file.10.1107/S2056989023003158/hb80582sup5.cmlSupporting information file. DOI: 2254180CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound exhibits I\u22ef\u03c0 halogen bonding and \u03c0-stacking in its extended structure. 18H8I2, is an ethynyl-substituted anthracene. The C\u2014C\u2014I bond angles deviate from 180\u00b0, being affected by inter\u00admolecular I\u22ef\u03c0 inter\u00adactions. These inter\u00adactions form a two-dimensional supra\u00admolecular structure further supported by offset \u03c0\u2013\u03c0 stacking of neighboring anthracene moieties.The title compound, C Two-dimensional crystals can have unique properties with applications in electronics, biomedicine, and sensors \u2014I moieties and the \u03c0-electrons of the adjacent anthracene rings [I1\u22efCg1 = 3.528\u2005(4)\u2005\u00c5 and C16\u2014I1\u22efCg1 = 151.2\u2005(3)\u00b0] and I2 has a short contact to the centroid of the C9\u2013C14 ring (Cg2) [I2\u22efCg2 = 3.767\u2005(4)\u2005\u00c5 and C18\u2014I2\u22efCg2 = 150.1\u2005(3)\u00b0]. The bent nature of the C\u2014I\u22efcentroid inter\u00adactions leads to short I\u22efC contacts ranging from 3.352\u2005(4) to 3.655\u2005(4)\u2005\u00c5. The shorter contact between I1 and Cg1 appears to influence more significantly the bending of the entire alkynyl substituent [C1\u2014C15\u2014I1 = 173.8\u2005(3)\u00b0 versus C8\u2014C17\u2014I2 = 178.7\u2005(3)\u00b0], notably pulling the I1 atom away from the central ring of the neighboring anthracene mol\u00adecule and toward its C2\u2013C7 centroid.The crystal stucture represents the first example of an ethynyl\u2013anthracene halogenated with iodine Fig.\u00a01. The C\u2014Igs Fig.\u00a02, where Iet al., 2008et al., 2014et al., 2015tert-but\u00adyl)[4-(iodo\u00adethyn\u00adyl)phen\u00adyl]carbamate plane Fig.\u00a03. Inter\u00adeet al., 2021N-iodo\u00adsuc\u00adcinimide and AgNO3 were added to dry di\u00admethyl\u00adformamide (5\u2005ml), and the resulting mixture was stirred under nitro\u00adgen. After 5\u2005h, the reaction mixture was diluted with EtOAc (30\u2005ml) and washed with H2O (5 \u00d7 30\u2005ml). The organic layer was dried in vacuo, resulting in an orange solid. The product was crystallized from the orange solid using vapor\u2013vapor diffusion (CH2Cl2/hexa\u00adnes).The procedure was modeled after an analogous functionalization of an alkynylsilane : \u03b4 8.33 , 7.82 .Crystal data, data collection, and structural refinement details are summarized in Table\u00a0110.1107/S2414314623005539/hb4433sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2414314623005539/hb4433Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623005539/hb4433Isup3.cmlSupporting information file. DOI: 2191397CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E-configuration with respect to the double bond of the hydrazone bridge and with an ac\u00adyl\u2013hydrazone (\u2014CH=N\u2014NH\u2014CO\u2014) torsion angle of 166.0\u2005(3)\u00b0. The mol\u00adecule exhibits a non-planar conformation, likely induced by packing requirements.The title aroylhydrazone ether exists in an 22H19N3O4, shows a non-coplanar conformation, with dihedral angles between the phenyl rings of 73.3\u2005(1) and 80.9\u2005(1)\u00b0. These deformations are induced by the crystal packing that is mainly governed by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming a mono-periodic arrangement parallel to the b axis.The mol\u00adecular structure of the title compound, C The active pharmacophore group, \u2014CH=N\u2014NH\u2014C=O\u2014, present in a hydrazone is primarily responsible for its broad spectrum of biological aspects \u2005\u00c5 is slightly shorter than that of 1.397\u2005(4)\u2005\u00c5 determined in the corresponding derivative having a thienyl ring replacing the p-nitro\u00adphenyl group , 3315 \u03bd(N\u2014H), 1606 \u03bd(C=Nazomethine). LC\u2013MS (FAB) m/z: [M + H]+ calculated for C22H19N3O4; 390.1446; found 390.1448.Yield: 0.29\u2005g, 79%; melting point (m.p.): 531\u2013533\u2005K; FT\u2013IR: 1636 \u03bd I, global. DOI: 10.1107/S2056989023003948/wm5682Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023003948/wm5682Isup3.cmlSupporting information file. DOI: 2232132CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E)-1-(4-bromo\u00adphen\u00adyl)-3-(2-methyl\u00adphen\u00adyl)prop-2-en-1-one, the mol\u00adecules are linked into chains by weak C\u2014H\u22efO inter\u00adactions along the b axis. Successive chains form a zigzag structure along the c axis, and these chains are connected to each other by face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions along the a axis, forming layers parallel to the (001) plane. The crystal structure maintains its stability via van der Waals inter\u00adactions between the layers.In the crystal of \u00b0, and the configuration about the C=C bond is E. In the crystal, the mol\u00adecules are linked into chains by weak C\u2014H\u22efO inter\u00adactions along the b axis. Successive chains form a zigzag structure along the c axis, and these chains are connected to each other by face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions along the a axis. These layers, parallel to the (001) plane, are linked by van der Waals inter\u00adactions, thus consolidating the crystal structure. Hirshfeld surface analysis showed that the most significant contacts in the structure are H\u22efH (43.1%), C\u22efH/H\u22efC (17.4%), Br\u22efH/H\u22efBr (14.9%), C\u22efC (11.9%) and O\u22efH/H\u22efO (9.8%).In the title com\u00adpound, C The mol\u00adecule is approximately planar, as indicated by the torsion angles C10\u2014C5\u2014C4\u2014C3 = 1.9\u2005(5)\u00b0, C9\u2014C4\u2014C3\u2014C2 = \u22124.4\u2005(5)\u00b0, C4\u2014C3\u2014C2\u2014C1 = \u2212176.3\u2005(3)\u00b0, C3\u2014C2\u2014C1\u2014C11 = \u2212168.2\u2005(3)\u00b0, C2\u2014C1\u2014C11\u2014C12 = 15.9\u2005(4)\u00b0 and Br1\u2014C14\u2014C15\u2014C16 = 178.5\u2005(2)\u00b0. The dihedral angle between the planes of the 2-methyl\u00adphenyl and 4-bromo\u00adphenyl rings is 23.49\u2005(15)\u00b0.The title com\u00adpound Fig.\u00a01 is com\u00adp3.C(5) chains \u2005\u00c5, slippage = 1.890\u2005\u00c5; Cg2\u22efCg2a = 3.9420\u2005(18)\u2005\u00c5, slippage = 1.942\u2005\u00c5; symmetry code: (a) x\u00a0\u2212\u00a01, y, z; Cg1 and Cg2 are the centroids of the 2-methyl\u00adphenyl (C4\u2014C9) and 4-bromo\u00adphenyl (C11\u2013C16) rings, respectively]. They form layers parallel to the (001) plane through van der Waals inter\u00adactions, thus consolidating the crystal structure.In the crystal, the mol\u00adecules are linked into is Fig.\u00a03 and thesCrystalExplorer17.5 ; 43.1%], C\u22efH/H\u22efC , Br\u22efH/H\u22efBr , C\u22efC and O\u22efH/H\u22efO inter\u00adactions contribute the most to the surface contacts. The crystal packing is additionally influenced by Br\u22efC/C\u22efBr (2.0%), Br\u22efBr (0.8%), N\u22efN (2.6%) and O\u22efC/C\u22efO (0.2%) contacts. The Hirshfeld surface study confirms the significance of H-atom inter\u00adactions in the packing formation. The large number of H\u22efH, C\u22efH/H\u22efC, Br\u22efH/H\u22efBr, C\u22efC and O\u22efH/H\u22efO inter\u00adactions indicates that van der Waals inter\u00adactions and hydrogen bonding are important in the crystal packing -1,3-di\u00adphenyl\u00adprop-2-en-1-one\u2019 unit in the Cambridge Structural Database \u2005\u00c5], forming layers lying parallel to the ab plane. In the crystal of RUCKIM, the mol\u00adecules are linked through type II halogen bonds, forming a sheet structure parallel to the bc plane. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions are observed between the sheets. In the crystal of XOLLOC, mol\u00adecules are linked via pairs of C\u2014H\u22efO inter\u00adactions with an via C\u2014H\u22ef\u03c0 inter\u00adactions. In the crystal of OBIYUW01, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions between the bromo\u00adphenyl and fluoro\u00adphenyl rings, resulting in a two-dimensional layered structure parallel to the ab plane. The mol\u00adecular packing is consolidated by weak Br\u22efH and F\u22efH contacts.In the crystal of KOCZUA, the shortest inter\u00admolecular contacts are Cl\u22efO [3.173\u2005(3)\u2005\u00c5]; these link the mol\u00adecules to form a 25.et al., 2021v/v) solution at room temperature.The title com\u00adpound was synthesized using a reported procedure (Chithiraikumar 6.Uiso(H) = 1.2Ueq(C) for aromatic H atoms, and C\u2014H = 0.98\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023007387/tx2073sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989023007387/tx2073Isup2.hklStructure factors: contains datablock(s) I. DOI: 2290092CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "This salt consists of a complex [Ni(OPD)2(H2O)2]2+ cation with two bidentate OPD ligands and trans aqua ligands, and a non-coordinating NDS2\u2013 anion, which is the double-deprotonated form of H2NDS. The NiII atom is situated at a center of inversion and exhibits a slightly tetra\u00adgonally distorted {O2N4} octa\u00adhedral coordination environment, with four shorter equatorial Ni\u2014N bonds [2.0775\u2005(17) and 2.0924\u2005(18)\u2005\u00c5] and a longer axial Ni\u2014O bond [2.1381\u2005(17)\u2005\u00c5]. The OPD ligand is located about an inversion center and is nearly coplanar with the NiN4 plane [dihedral angle 0.95\u2005(9)\u00b0]. In the crystal, the cations and anions are connected by charge-assisted inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efO hydrogen-bonding inter\u00adactions into the tri-periodic network structure. Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from H\u22efH (44.1%), O\u22efH/H\u22efO (34.3%), C\u22efH/H\u22efC (14.8%) C\u22efC (6.5%) (involving the cations) and O\u22efH/H\u22efO (50%), H\u22efH (25%), C\u22efH/H\u22efC (15.3%), C\u22efC (8.2%) (involving the anions) inter\u00adactions.The reaction of Its reactions with carb\u00adoxy\u00adlic acids and their derivatives produce the important class of benzimidazoles or its deprotonated form (sulfonates) are of inter\u00adest in supra\u00admolecular chemistry 2]\u00b7NDS, where the NDS2\u2013 anion is not part of the metal coordination sphere.In this work, we focus on the synthesis, crystal structure, and Hirshfeld surface analysis of a nickel(II) complex, and a pair of longer axial Ni\u2014O bonds [2.1381\u2005(17)\u2005\u00c5]. The OPD ligand, likewise located over a crystallographic inversion center at the middle of the central C11\u2014C11 bond, is almost coplanar with the NiN4 plane, with a dihedral angle of 0.95\u2005(9)\u00b0. The deviation of the ideal octa\u00adhedral coordination sphere around nickel might be explained as follows: The inflexible nature of the OPDA ring system with an N\u22efN distance between the amino groups of 2.770\u2005(2)\u2005\u00c5 determines the N2\u2014Ni1\u2014N1 and N2\u2014Ni1\u2014N1 bite angles of 83.26\u2005(7) and 96.74\u2005(7) \u00b0, respectively.The structures of the mol\u00adecular entities of the title compound are shown in Fig.\u00a013.2(H2O)2]2+ cation and the NDS2\u2013 anion are associated via charge-assisted inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds (Table\u00a012(H2O)2]2+ cation forms N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds with six neighboring organic anions whereby the two aqua and two OPD ligands act solely as hydrogen-bonding donor groups . Additionally, cations and neighboring dianions are linked through O1W\u2014H1WA\u22efO3iv and N2\u2014H2B\u22efO3i hydrogen bonds. The mol\u00adecules stack along [001], thereby forming a consolidated tri-periodic supra\u00admolecular network s Table\u00a01. Each 2+ cation and the NDS2\u2013 dianion. The dnorm surface has twelve bright-red spots on the Hirshfeld surface for the cation and anion each 2]2+ cation and NDS2\u2013 anion, respectively; Fig.\u00a06b and 6f). O\u22efH/H\u22efO and C\u22efH/H\u22efC, inter\u00adactions in the cation, and H\u22efH and C\u22efH/H\u22efC inter\u00adactions in the dianion follow with contributions of 34.3, 14.8, 25 and 15.3%, respectively . Other minor contributions are from C\u22efC (6.5%) and C\u22efO (0.3%) contacts in the cation, and from C\u22efC (8.2%), C\u22efO (0.3%), O\u22efO (0.1%) and S\u22efH (0.1%) contacts in the dianion. The O\u22efH/H\u22efO contacts are visible as a spike with a sharp tip on the side of the corresponding two-dimensional fingerprint plot, which is indicative of strong inter\u00admolecular inter\u00adactions between atoms. On the other hand, the C\u22efH/H\u22efC contacts form less pronounced spikes, suggesting that these inter\u00adactions are much weaker.The supra\u00admolecular inter\u00adactions discussed above were qu\u00adanti\u00adtatively investigated and visualized using Hirshfeld surface analysis performed with ch Fig.\u00a05, resultily Fig.\u00a06c,d,g,h.4.et al., 2016o-phenyl\u00adenedi\u00adamine moiety were identified. Out of these, 129 compounds were metal complexes, while 78 compounds were organic salts. One organic salt comprising protonated o-phenyl\u00adenedi\u00adamine and 1,5-naphthalene\u00addisulfonate has been studied sulfate hepta\u00adhydrate and disodium naphthalene-1,5-di\u00adsulfonate in 10\u2005ml of the same mixed ethanol/water solvent. The resulting mixture was heated under reflux and stirred for 40\u2005min. After 5\u2005d of slow solvent evaporation at room temperature, a light-green crystalline product was obtained with a yield of 65% (based on Ni). Elemental analysis calculated (%) for C22H26N4NiO8S2: C 44.24, H 4.39, N 9.38; found: C 44.18, H 4.34, N 9.31.The starting materials are commercially available and were used without further purification. The ligand OPDA was dissolved in 10\u2005ml of a 1:1 6.Uiso(H) = 1.2Ueq(C) and C\u2014H = 0.93\u2005\u00c5 for aromatic H atoms. Hydrogen atoms of the amino groups and of the water mol\u00adecule were located using a difference-Fourier map and refined with bond-length restraints of 0.89\u2005(1) and 0.85\u2005(1)\u2005\u00c5, respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023009350/wm5701sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023009350/wm5701Isup2.hklStructure factors: contains datablock(s) I. DOI: 2303464CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Bombina maxima. Here, we reported that B.\u00a0maxima cells secreted \u03b2\u03b3-CAT under glucose, glutamine, and pyruvate deficiency to scavenge extracellular proteins for their nutrient supply and survival. AMPK signaling positively regulated the expression and secretion of \u03b2\u03b3-CAT. The PFP complex selectively bound extracellular proteins and promoted proteins uptake through endolysosomal pathways. Elevated intracellular amino acids, enhanced ATP production, and eventually prolonged cell survival were observed in the presence of \u03b2\u03b3-CAT and extracellular proteins. Liposome assays indicated that high concentration of ATP negatively regulated the opening of \u03b2\u03b3-CAT channels. Collectively, these results uncovered that \u03b2\u03b3-CAT is an essential element in cell nutrient scavenging under cell nutrient deficiency by driving vesicular uptake of extracellular proteins, providing a new paradigm for PFPs in cell nutrient acquisition and metabolic flexibility.Nutrient acquisition is essential for animal cells. \u03b2\u03b3-CAT is a pore-forming protein (PFP) and trefoil factor complex assembled under tight regulation identified in toad \u2022The PFP \u03b2\u03b3-CAT is expressed and secreted by toad cells under nutrient deficiency\u2022AMPK signaling positively regulates the expression and secretion of \u03b2\u03b3-CAT\u2022The secreted \u03b2\u03b3-CAT selectively binds and scavenges extracellular nutrients\u2022The high concentrations of ATP may negatively regulate \u03b2\u03b3-CAT channel opening Cell biology The former operates in uptake of small nutrient, such as glucose and amino acids, and the latter is the main pathway through which cells obtain insoluble nutrients, such as cholesterol and iron.,,Bombina maxima. It forms membrane pores (channels) with a functional diameter 1.5\u20132.0\u00a0nm.,,,,,Numerous pore-forming proteins (PFPs) with a membrane insertion domain similar to bacterial toxin aerolysin, namely aerolysin family PFPs , have been found in plants and animals.,,,,,This PFP complex firstly targets cell surface acidic glycosphingolipids in lipid rafts via a double-receptor binding model, corresponding to BmALP1 subunit and BmTFF3 subunit binding gangliosides and sulfatides, respectively.B.\u00a0maxima cells to further investigate the role of SELC protein \u03b2\u03b3-CAT in cell nutrient acquisition and metabolic flexibility under cell nutrient deficiency. Interestingly, toad liver and gastrointestinal cells secreted \u03b2\u03b3-CAT to scavenge extracellular protein nutrients, such as albumin and ovalbumin, through endolysosomal pathways in the absence of glucose, glutamine and pyruvate, which elevated intracellular amino acids and ATP levels and supported cell survival under nutrient deficiency. The expression and secretion of \u03b2\u03b3-CAT were largely attenuated by inhibition of AMP-activated kinase (AMPK) signaling. Furthermore, in a liposome model, ATP, but not AMP inhibited \u03b2\u03b3-CAT channel opening. These results revealed the essential role of a secretory PFP in extracellular nutrient scavenging by cells through endolysosomal pathways under cell nutrient deficiency.Here, we used the nutrient deficiency model of toad ,,,,+/Gln+/Pyr+), glucose/glutamine/pyruvate-depleted medium (Glc\u02c9/Gln\u02c9/Pyr\u02c9), and glucose-containing but glutamine/pyruvate-depleted medium (Glc+/Gln\u02c9/Pyr\u02c9). In the isolated toad liver cell population, there were 62.4% hepatocytes on the basis of toad skin transcriptomeT172 and pACC1S80 in human, respectively) were evolutionarily conserved from toad B.\u00a0maxima to human in culture supernatants of toad liver cells under Glc\u02c9/Gln\u02c9/Pyr\u02c9 conditions , and dextran (70\u00a0kDa), an indicator of macropinocytosisKD values of \u03b2\u03b3-CAT with OVA and Bm-SA was approximately 2.33\u00a0\u00d7\u00a010\u22128\u00a0M and 2.47\u00a0\u00d7\u00a010\u22128\u00a0M, respectively. Ovalbumin-DQ (OVA-DQ) is a fluorescent indicator that fluoresces on proteolytic degradation.\u02c9/Gln\u02c9/Pyr\u02c9 conditions was observed by scanning confocal microscopy under Glc\u02c9/Gln\u02c9/Pyr\u02c9 conditions , toad croscopy B and flocroscopy C. \u03b2\u03b3-CATs at 3\u00a0h A, a leves \u03b2\u03b3-CAT B and 3C.nditions B. Furthenditions D. Ethyl-nditions C and mamnditions D in the nditions E. Collec\u02c9/Gln\u02c9/Pyr\u02c9 conditions for 7\u00a0h by LC/MS and LC-MS/MS. The levels of amino acids, including asparagine and glutamine, in toad liver cells treated with bovine serum albumin (BSA) and \u03b2\u03b3-CAT were augmented compared with those in cells treated with BSA only staining.nditions A. Notablnditions A. Howevenditions A or ovalnditions B. Additinditions C and 4D.\u02c9/Gln\u02c9/Pyr\u02c9 conditions on liposomes, which induces dye release from lipid vesicles, an advantageous model without ATP receptors.nditions C. Notablnditions D. This rnditions E. Collec,,B.\u00a0maxima PFP protein complex \u03b2\u03b3-CAT in cell nutrient scavenging and energy supply under the deprivation of glucose and glutamine, essential nutrient components in cell metabolism.B.\u00a0maxima \u03b2\u03b3-CAT indeed acts as a novel system driven cell vesicular delivery that should play a physiological role in cell nutrient acquisition by mediating cellular nutrient import through endolysosomal pathways.,,PFPs are widely distributed in all kingdoms of life, which have long been recognized as either pore-forming toxin for microbial infection or host immune executors.+/Gln+/Pyr+) from 1 to 5\u00a0h and intracellular nutrient sensors, which survey the abundance of energy and major metabolites, play an important role in metabolic homeostasis and cell survival.,,,,in\u00a0vivo and in\u00a0vitro in toad osmoregulatory organs to facilitate toad water maintaining.in\u00a0vivo. Indeed, the secreted \u03b2\u03b3-CAT could bind to ovalbumin and toad B.\u00a0maxima serum albumin (nutrients) directly. Furthermore, the colocalization of \u03b2\u03b3-CAT and OVA-DQ was readily observed under cell nutrient deficiency. These results were suggested that the pathway of \u03b2\u03b3-CAT importing and enriching nutrients might be selective, which was different from classical pinocytosis/macropinocytosis-like endocytosis.Diverse endocytic pathways are available at the surface of metazoan cells.,,,The functional diameter of bacterial toxin aerolysin channels is approximately 1.5\u00a0nm, which is large enough for translocation of oligonucleotides, peptides, and unfolded proteins.,,,,The physiological concentrations of ATP in cells are estimated to be 5\u201310\u00a0mM.,\u03b2\u03b3-CAT is a complex of BmALP1 and BmTFF3, in which BmTFF3 acts as a chaperon and regulatory unit of BmALP1 to stabilize the PFP monomer and deliver it to proper targets.,,,It is well documented that autophagy is a cellular process to sequester and degrade intracellular components under nutrient deficiency, which is the last defense for cells under nutrient deficiency and excess autophagy may lead to cell death.B.\u00a0maxima cells secrete \u03b2\u03b3-CAT to scavenge extracellular nutrients under nutrient deficiency at the cellular level (present study). Second, \u03b2\u03b3-CAT in toad B.\u00a0maxima blood circulation is an immediate and active responsive element under toad fasting in\u00a0vivo, which trans cellularly deliver and transport albumin-bound fatty acids to tissue parenchymal cells for their nutrient supply.B.\u00a0maxima physiology for adaptation to various nutrient environments. Rationally, similar strategies and executive pathways should be conserved in vertebrates, in which various families of PFPs including af-PFPs are widely distributed. Knowledge from \u03b2\u03b3-CAT can provide clues to understand novel PFP-driven cell vesicular delivery systems in nutrient acquisition and metabolic flexibility. Although af-PFPs have not been clearly observed in Eutherian mammals, other PFP family members can readily compensate for the role of af-PFPs.SELC protein \u03b2\u03b3-CAT works in cell nutrient acquisition at least at two levels. First, toad B.\u00a0maxima cells secrete \u03b2\u03b3-CAT, a PFP and trefoil factor (TFF) complex assembled depending on environmental cues under glucose, glutamine, and pyruvate deficiency. This PFP complex supports cell survival by driving the cellular import of extracellular proteins through endolysosomal pathways. The imported proteins serve as nutrients in nutrient-deprived cells for energy supply. AMPK signaling positively regulates the expression and secretion of \u03b2\u03b3-CAT, whereas high concentrations of ATP (> 1\u00a0mM) bind to and negatively regulate \u03b2\u03b3-CAT channels. Our findings define the essential role of toad B.\u00a0maxima PFP complex \u03b2\u03b3-CAT in cell macromolecular nutrient scavenging, providing a new paradigm for PFPs in cell nutrient acquisition and metabolic flexibility.In conclusion, the present study elucidated that toad in\u00a0vitro cell experiments. Consequently, it is necessary to study whether the secretory PFP \u03b2\u03b3-CAT contribute to the nutrient acquisition in\u00a0vivo under starvation, which revealed that the PFP \u03b2\u03b3-CAT was secreted into toad blood in response to toad fasting,,,,Our work was restricted zhangy@mail.kiz.ac.cn).Further information and requests for resources and reagents should be directed to and will be fulfilled by the lead contact, Yun Zhang (All unique/stable reagents generated in this study are available from the B.\u00a0maxima) was performed as described previously and the males with a mean body weight of 25\u00a0\u00b1 5\u00a0g were used for this study . All procedures and the care and handing of animals were approved by the Ethics Committee of the Kunming Institute of Zoology, Chinese Academy of Sciences .Feeding of toads containing 10% fetal bovine serum and 1% Penicillin-Streptomycin Solution at 37\u00b0C with 5% CO+/Gln+/Pyr+ medium, Biological Industries, Cat 01-172-1A), glucose, pyruvate and glutamine-free medium , or added 3.151\u00a0g/L glucose to Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium (Glc+/Gln\u02c9/Pyr\u02c9 medium). For macropinocytosis inhibition, cells were first incubated with 100\u00a0\u03bcM EIPA (5-(N-ethyl-N-isopropyl)-Amiloride), a Na+/H+-exchanger inhibitor used for macropinocytosis inhibition) in Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium for 1\u00a0h at 26\u00b0C (toad cells) or 37\u00b0C (HepG2 cells), respectively. For AMPK signaling inhibition, cells were treated with 0\u201310\u00a0\u03bcM compound C or 0\u201320\u00a0\u03bcM SBI-0206965 in Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium for 3\u00a0h at 26\u00b0C. To deplete endogenous \u03b2\u03b3-CAT, toad cells were incubated with 100\u00a0\u03bcg/mL anti-\u03b2\u03b3-CAT rabbit polyclonal antibodies or 100\u00a0\u03bcg/mL rabbit IgG as the isotype control in Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium at 26\u00b0C.Cells were cultured in basal medium containing 3.151\u00a0g/L glucose, 1\u00a0mM pyruvate and 2.5\u00a0mM glutamine according to the manufacturer\u2019s instructions.Total 2\u00d7106 isolated toad liver cells were cultured at 26\u00b0C in various media for 0, 3, 5 and then up to 11\u00a0h at 2-h intervals. The treatments were as follows: Glc+/Gln+/Pyr+ medium, Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium, Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium containing 100\u00a0nM \u03b2\u03b3-CAT, Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium containing 500\u00a0\u03bcg/mL ovalbumin or BSA , Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium containing 100\u00a0nM \u03b2\u03b3-CAT and 500\u00a0\u03bcg/mL OVA or BSA, Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium containing 100\u00a0\u03bcg/mL anti-\u03b2\u03b3-CAT antibodies in the presence of 500\u00a0\u03bcg/mL OVA or BSA, and Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium containing 100\u00a0\u03bcg/mL rabbit IgG in the presence of 500\u00a0\u03bcg/mL OVA or BSA. To assess the viability of toad stomach cells, 5\u00d7106 isolated toad stomach cells were subjected to various treatments for 7\u00a0h at 26\u00b0C. To assess cytotoxicity of compound C and SBI-0206965, 5\u00d7106 isolated toad liver cells were cultured in Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium plus compound C (0\u201310\u00a0\u03bcM) or SBI-0206965 (0\u201320\u00a0\u03bcM) for 3\u00a0h at 26\u00b0C. After treatments, toad cells were stained with 500\u00a0ng/mL PI for 4\u00a0min at 26\u00b0C. Fluorescence was recorded using an LSR Fortessa cell analyzer .Cell viability of toad cells was measured using propidium iodide (PI) stain assay as described previously.6 toad liver cells were cultured in Glc+/Gln+/Pyr+ medium or Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium containing various concentration of \u03b2\u03b3-CAT (0\u20131000\u00a0nM) for 3\u00a0h, then the supernatant of toad cells was collected, and LDH release was detected by LDH cytotoxicity assay kit according to the manufacturer\u2019s instructions.To assess the cytotoxicity of \u03b2\u03b3-CAT in toad liver cells, the total 1\u00d7104 cells were seeded into each well of a 96-well culture plate. After overnight culture, the cells were washed thrice with PBS and incubated in Glc+/Gln+/Pyr+ medium, Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium, Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium plus various concentrations of \u03b2\u03b3-CAT (0\u2013160\u00a0nM), or Glc\u02c9/Gln\u02c9/Pyr\u02c9 medium plus 5\u00a0mg/mL OVA in the presence or absence of 40\u00a0nM \u03b2\u03b3-CAT for 36\u00a0h. Then, the cells were incubated with MTS reagent for 2\u00a0h in the dark at 37\u00b0C with 5% CO2. Absorbance was read at 490\u00a0nm with an Infinite 200 Pro microplate reader .To assess viability of mammalian HepG2 cells, 1\u00d7107 toad liver, intestinal, or stomach cells was collected and then washed twice with 30\u00a0mL Ringer\u2019s solution. Toad liver cells were cultured in Glc+/Gln+/Pyr+, Glc\u02c9/Gln\u02c9/Pyr\u02c9, or Glc+/Gln\u02c9/Pyr\u02c9 medium for 1\u20135\u00a0h at 26\u00b0C, and intestinal or stomach cells was cultured for 1\u00a0h at 26\u00b0C. Then, culture supernatants were collected and prepared for the hemolytic activity assay as described previously.A total of 1\u00d710in\u00a0vitro, and the detail methods were performed as described previouslyKD were determined using Octet Analysis Studio Software using a 1:1 model.The BLI assay was used for the interaction between \u03b2\u03b3-CAT and the extracellular nutrients 6 isolated toad liver cells were treated as described in the \u201cB.\u00a0maxima serum albumin ) or 20\u00a0\u03bcg/mL Ovalbumin-DQ in the dark at 26\u00b0C for 15\u00a0min. Fluorescence was detected by the LSR Fortessa cell analyzer using the FITC channel. In each sample, 1\u00d7104 single cells were analyzed.A total of 5\u00d710For immunofluorescence, cells were incubated with 100\u00a0\u03bcg/mL Ovalbumin-DQ in the dark at 26\u00b0C for 15\u00a0min. To stain \u03b2\u03b3-CAT, cells were washed and incubated with 20\u00a0\u03bcg/mL anti-\u03b2\u03b3-CAT primary antibodies overnight at 4\u00b0C in the dark. After washing with PBS three times, the cells were incubated with 10\u00a0\u03bcg/mL cy3-conjugated anti-rabbit IgG for 1\u00a0h at 37\u00b0C in the dark. Then, the samples were sealed with an anti-fluorescent quench agent containing DAPI. Images were acquired by a Zeiss LSM 880 microscope system .7 isolated toad liver cells were treated in various media as described in the \u201c2O and MeOH solution (1:4) at\u00a0\u221280\u00b0C, and then the samples were incubated for 15\u00a0min at \u221280\u00b0C. Then, the supernatants were collected by centrifuging at 17,000\u00a0g for 10\u00a0min 10\u00a0\u03bcL of these supernatants were injected per analysis. Samples were run on a Vanquish (Thermo Fisher Scientific) UHPLC system with mobile phase A and mobile phase B (ACN) at a flow rate of 200\u00a0\u03bcL/min on an Accucore-150-Amide-HILIC column 2.6\u00a0\u03bcm (100\u00a0\u00d7\u00a02.1\u00a0mm) at 40\u00b0C at a gradient from 45% to 90% A in 15\u00a0min followed by a 10-min isocratic step. The UHPLC was coupled to a Q-Exactive (Thermo Fisher Scientific) mass analyzer running in polarity switching mode at 3.5\u00a0kV for positive change scanning and 3.0\u00a0kV for negative change scanning at an MS1 resolution of 70,000. Metabolites were identified by exact mass (MS1), retention time, and in some cases, by their fragmentation patterns (MS2) at normalized collision energy. Quantification was performed by area under the curve integration of MS1 ion chromatograms with the Thermo Scientific Xcalibur software package. Area values were normalized to cell count averages from triplicate wells treated in parallel for each condition.A total of 1\u00d710Intracellular ATP was extracted and measured by an ATP detection kit in accordance with the manufacturer\u2019s protocol. Briefly, toad liver cells were treated same as described in the \u201cTo assess the effect of adenosine phosphate on \u03b2\u03b3-CAT channel formation-induced liposome dye release, liposomes were incubated with 600\u00a0nM \u03b2\u03b3-CAT in the presence or absence of adenosine phosphate (Sigma), and then dye release assays were performed as described previously.The purification of \u03b2\u03b3-CAT were performed as described previously.KD) was determined using BIA evaluation 4.1 software and its equation for 1:1 Langmuir binding.A direct interaction was assessed as described previously.+/Gln+/Pyr+, Glc\u02c9/Gln\u02c9/Pyr\u02c9, or Glc+/Gln\u02c9/Pyr\u02c9 medium, respectively. Then, the cells were lysed and prepared for Western blotting. The cell supernatant was concentrated to one-10th of the original volume by a vacuum lyophilizer for Western blotting as described previously.To assess protein level of \u03b2\u03b3-CAT in toad liver cell lysate and supernatant, cells were cultured in GlcTo assess AMP-activated protein kinase (AMPK) signaling, cells were treated as described in the \u201cThe mRNA levels of BmALP1 (\u03b2\u03b3-CAT-\u03b1) and BmTFF3 (\u03b2\u03b3-CAT-\u03b2) in toad liver cells were measured by quantitative real-time PCR (qRT-PCR) using a ChamQ Universal SYBR qPCR Master Mix kit . The cycle counts of target genes were normalized to those of \u03b2-actin. Primer sequences of \u03b2\u03b3-CAT and \u03b2-actin used were the same as report previously.2, and 1\u00a0mM EDTA, pH 7.5) for 30\u00a0min at 26\u00b0C. Cell lysates from HepG2 cells were used as a positive control. Absorbance was read at 620\u00a0nm, and the free phosphate concentration was calculated by a standard curve (0\u201350\u00a0\u03bcM free phosphate).The ATPase activity of \u03b2\u03b3-CAT was measured by an ATPase activity assay kit according to the manufacturer\u2019s protocol. Briefly, various concentrations \u03b2\u03b3-CAT was incubated with 4\u00a0mM ATP in assay buffer (40\u00a0mM Tris, 80\u00a0mM NaCl, 8\u00a0mM MgAcB.\u00a0maxima with other species was conducted by Clustal Omega6.Sequence alignment analysis of AMPK\u03b1 and ACC1 in t-test. Multiple comparisons were performed by one-way ANOVA with post-hoc contrasts by Dunnett\u2019s multiple comparisons test. Multi-group comparisons were made by two-way ANOVA with Sidak\u2019s multiple comparisons test. p < 0.05 was considered statistically significant. the statistical details of experiments can be found in the figure legends.All experimental values are expressed as means\u00a0\u00b1 SD. Each experiment was repeated at least twice. All data were analyzed using GraphPad Prism 8.0 software. Two comparisons were performed using the standard unpaired"} +{"text": "The low-temperature (90\u2005K) crystal structure of 4-(di\u00admethyl\u00adaza\u00adnium\u00adyl)-2-hy\u00addroxy\u00adanilinium dichloride monohydrate is presented along with a Hirshfeld surface analysis of the organic cation. 8H14N2O+\u00b72Cl\u2212\u00b7H2O, at low temperature (90\u2005K) are presented. The organic cation is essentially planar: the r.m.s. deviation of its non-hydrogen atoms (aside from the two methyl groups) is 0.0045\u2005\u00c5. The methyl carbons are 1.3125\u2005(12)\u2005\u00c5 and 1.1278\u2005(12)\u2005\u00c5 either side of the mean plane. The crystal packing involves extensive hydrogen bonding of types O\u2014H\u22efCl, N\u2014H\u22efCl, N\u2014H\u22efOW, and OW\u2014HW\u22efCl (where W = water), which arrange into chains of R24(12) motifs that combine to form corrugated layers parallel to -2-hy\u00addroxy\u00adanilinium dichloride monohydrate, C Perkin, 1896aka, acetamino\u00adphen/Tylenol) and the fenamate family of NSAIDs (anthranilic acid deriv\u00adatives). Within this context, a concise review of aniline and its derivatives was presented by Anjalin et al. , at 90\u2005K.Given the industrial and pharmaceutical significance of anilinium salts, this paper presents the crystal structure and Hirshfeld-surface analysis of 4-(di\u00admethyl\u00adaza\u00adnium\u00adyl)-2-hy\u00addroxy\u00adanilinium dichloride monohydrate and N2\u2014H2N\u22efCl2 [D\u2013Ad = 3.0467\u2005(9)\u2005\u00c5] hydrogen bonds with the chloride anions, which in turn act as acceptors for O1W\u2014H1W1\u22efCl1vi [D\u2013Ad = 3.1493\u2005(9)\u2005\u00c5] and O1W\u2014H2W1\u22efCl2vi [D\u2013Ad = 3.1036\u2005(9)\u2005\u00c5] hydrogen bonds with the water mol\u00adecule (symmetry codes as per Table\u00a01via N1\u2014H3N1\u22efO1W [D\u2013Ad = 2.7093\u2005(12)\u2005\u00c5] hydrogen bonds, forming chains that extend parallel to [101] \u2005\u00c5] and N1\u2014H1N1\u22efCl2i [D\u2013Ad = 3.1299\u2005(9)\u2005\u00c5] hydrogen bonds, forming corrugated layers parallel to Fig.\u00a03.normd for the cation in I were obtained using CrystalExplorer 4+\u00b74Cl\u2212\u00b74H2O; Stylianou et al., 2017I. Two other anilinium salts not returned in the above search but that share similar features to I are POMXUL and PAXXIX -(hy\u00addroxy\u00adimino)\u00admeth\u00adyl]-N,N-dimethyl anilinium chloride (C9H13N2O+\u00b7Cl\u2212).A search of the Cambridge Structural Database and 0.98\u2005\u00c5 (RCH3) and Uiso(H) parameters set to either 1.2Ueq (R2CH) or 1.5Ueq (RCH3) of the attached carbon. Nitro\u00adgen and oxygen-bound hydrogens were fully refined .Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023007223/vm2289sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989023007223/vm2289Isup2.hklStructure factors: contains datablock(s) I. DOI: 2289098CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "IO2\u00a0=\u00a00.209; HIGH FIO2\u00a0=\u00a00.155; VHIGH FIO2\u00a0=\u00a00.125) on glycaemic control, insulin sensitivity, and oxidative stress during a subsequent oral glucose tolerance test (OGTT) in males with overweight (mean (SD) BMI\u00a0=\u00a027.6 (1.3) kg/m2; n\u00a0=\u00a012). Feasibility was defined by exceeding predefined withdrawal criteria for peripheral blood oxygen saturation (SpO2), partial pressure of end\u2010tidal oxygen or carbon dioxide and acute mountain sickness (AMS), and dyspnoea symptomology. Hypoxia reduced SpO2 in a stepwise manner (CON\u00a0=\u00a097(1)%; HIGH\u00a0=\u00a091(1)%; VHIGH\u00a0=\u00a081(3)%, p\u2009<\u20090.001), but did not affect peak plasma glucose concentration (CON\u00a0=\u00a07.5(1.8) mmol\u2219L\u22121; HIGH\u00a0=\u00a07.7(1.1) mmol\u2219L\u22121; VHIGH\u00a0=\u00a07.7(1.1) mmol\u2219L\u22121; p\u00a0=\u00a00.777; \u03b72\u00a0=\u00a00.013), plasma glucose area under the curve, insulin sensitivity, or metabolic clearance rate of glucose (p\u2009>\u20090.05). We observed no between\u2010conditions differences in oxidative stress (p\u2009>\u20090.05), but dyspnoea and AMS symptoms increased in VHIGH (p\u2009<\u20090.05), with one participant meeting the withdrawal criteria. Acute HIGH or VHIGH exposure prior to an OGTT does not influence glucose homeostasis in males with overweight, but VHIGH is associated with adverse symptomology and reduced feasibility.Previous research has shown that \u226460\u2009min hypoxic exposure improves subsequent glycaemic control, but the optimal level of hypoxia is unknown and data are lacking from individuals with overweight. We undertook a cross\u2010over pilot feasibility study investigating the effect of 60\u2010min prior resting exposure to different inspired oxygen fractions (CON F Likewise, Mackenzie et al.\u00a0 were reduced, and insulin sensitivity improved, during a subsequent intravenous glucose tolerance test. In men with obesity (mean(SD) body mass index (BMI)\u00a0=\u00a032.7(1.3) kg/m2), a more prolonged intervention consisting of ten nights of moderate normobaric hypoxia (FIO2\u00a0=\u00a00.15) reduced fasting blood [glucose] and improved insulin sensitivity and two normobaric\u2010hypoxic conditions equivalent to a simulated altitude of ~2500\u2009m, i.e., \u2018high\u2019 altitude , and\u2009~\u20094000\u2009m, i.e., \u2018very\u2010high\u2019 altitude was recruited from the host university (mean (SD) age\u00a0=\u00a032(7) years; height\u00a0=\u00a01.82(0.08) m; mass\u00a0=\u00a091.8(10.1) kg; BMI\u00a0=\u00a027.6(1.3) kg/m2). Although intended as a pilot feasibility\u2010study, our sample size was in keeping with similar previous work that informed the study of 0.14 for a given outcome variable using repeated measures ANOVA . All participants were free from metabolic disease including T2DM (glycated hemoglobin (HbA1c)\u00a0=\u00a034(4) mmol/mol), thalassemia, and haemoglobinopathies, were not taking any medications and had normal resting lung function , ECG, and blood pressure.To balance the three conditions, a convenience sample of 12 males with overweight and connected to a metabolic cart calibrated using known concentrations of O2, CO2, and N2 gas, and a 3\u2010L air syringe . Participants were blind to the FIO2 on each occasion with CON serving as a sham control condition. Pulmonary gas exchange and ventilation were averaged into 60\u2010s time bins. Peripheral oxygen saturation was measured by fingertip pulse oximetry interfaced with a data acquisition system and heart rate using short range telemetry . Participants were withdrawn if their peripheral blood oxygen saturation (SpO2) fell below 65% for 15 continuous seconds, or if the partial pressure of end\u2010tidal oxygen (PETO2) fell below 45\u2009mmHg, or the partial pressure of end\u2010tidal carbon dioxide (PETCO2) fell below 25\u2009mmHg, for three consecutive breaths. Preliminary data collection indicated that, for most participants, the lower FIO2 would approximate the exposure limits permitted within our laboratory.Participants reported to the laboratory at 0730\u2009hours on test days following a 12\u2010h overnight fast and in a hydrated state, having only consumed water and having abstained from alcohol and strenuous exercise in the previous 24\u2009h. Participants rested in a semi\u2010recumbent position and a venous cannula was inserted into a forearm vein. Thereafter, following a 15\u2010min rest period, baseline measurements were obtained and the participant donned an oronasal mask delivering the required FInformed by previous studies showing beneficial effects of acute hypoxia on subsequent glycaemic control were removed. Energy expenditure and substrate utilization were calculated during the OGTT using indirect respiratory calorimetry, and hunger scores were summed over the OGTT period to create a single hunger score. Criteria for feasibility were (i) withdrawal from the study due to physiological perturbation exceeding any of the study withdrawal criteria for SpO2, PETO2, or PETCO2 or (ii) symptoms attributable to acute hypoxia (AMS and dyspnoea). Between\u2010conditions differences were examined using one\u2010way RM\u2010ANOVA, with the exception of AMS symptoms and perceived dyspnoea, which were analyzed using Friedman's test; if the RM\u2010ANOVA violated the assumption of sphericity, the Greenhouse\u2013Geisser statistic was employed. The effect size for RM ANOVA is reported as \u03b72, and post\u2010hoc analysis of significant RM\u2010ANOVA and Friedman's test effects were undertaken using (least significant difference) pairwise comparisons and Wilcoxon's signed ranks test, respectively.Statistical analyses were undertaken using SPSS Version 28 , and significance was set a priori at P\u2009\u2264\u20090.05. Data are presented mean (SD), with the exception of the Lake Louise questionnaire and dyspnoea scale data, which are presented as median(range). Peak and AUC (trapezoid method) were calculated for [glucose] and AOPP to provide an index of glucose homeostasis and a marker of oxidative stress, respectively, following the hypoxic exposure. Post\u2010prandial insulin sensitivity and metabolic clearance rate of glucose (a maker of insulin sensitivity) were calculated according to validated equations incorporating demographic data (age and BMI) and on the basis of [glucose] and [insulin] samples at 0, 60, and 120\u2009min of the OGTT and 0.126(0.05) for the HIGH and VHIGH trials, respectively. SpO2 differed between all conditions (CON\u00a0=\u00a097(1)%; HIGH\u00a0=\u00a091(1)%; VHIGH\u00a0=\u00a081(3)%; p\u2009<\u20090.001, \u03b72\u00a0=\u00a00.957) being highest in CON and lowest in VHIGH .One participant was unable to complete the experimental trials due to reaching the study P\u22121; HIGH\u00a0=\u00a07.7(1.1) mmol\u2219L\u22121; VHIGH\u00a0=\u00a07.7(1.1) mmol\u2219L\u22121; p\u00a0=\u00a00.777, \u03b72\u00a0=\u00a00.013) during the OGTT nor the area under the curve (CON\u00a0=\u00a011.5(2.9) mmol\u2219h\u22121/L; HIGH\u00a0=\u00a012.3(1.6) mmol\u2219h\u22121/L; VHIGH\u00a0=\u00a012.5(2.0) mmol\u2219h\u22121/L; p\u00a0=\u00a00.241, \u03b72\u00a0=\u00a00.146) differed between conditions \u03bcmol\u2219kg\u22121\u2219min\u22121\u2219pmol/L; VHIGH\u00a0=\u00a00.11(0.01) \u03bcmol\u2219kg\u22121\u2219min\u22121\u2219pmol/L; p\u00a0=\u00a00.351, \u03b72\u00a0=\u00a00.099) mL\u2219kg\u22121\u2219min\u22121; VHIGH\u00a0=\u00a09.6(0.7) mL\u2219kg\u22121\u2219min\u22121; p\u00a0=\u00a00.256, \u03b72\u00a0=\u00a00.142) were unaffected by the FIO2. Oxidative stress (plasma [AOPP]) before and after the hypoxic intervention period, and during the subsequent OGTT, is shown in Figure\u00a0p\u00a0=\u00a00.621, \u03b72\u00a0=\u00a00.051) and accumulated oxidative stress (AUC [AOPP]: CON\u00a0=\u00a0526(202) pmol\u2219h\u22121/L; HIGH\u00a0=\u00a0532(176) pmol\u2219h\u22121/L; VHIGH 567(203) pmol\u2219h\u22121/L; p\u00a0=\u00a00.606, \u03b72\u00a0=\u00a00.054) did not differ between the conditions, whereas IL\u20106 remained below the detectable limits of the assay (9.4\u00a0pg\u2219mL\u22121).Plasma [glucose] and [insulin] before and after the hypoxic intervention period, and during the subsequent OGTT, are shown in Figure\u00a0p\u00a0=\u00a00.011, \u03b72\u00a0=\u00a00.394), being lower in HIGH than VHIGH or CON (p\u2009<\u20090.05), although at an individual substrate level neither the total energy from fat (CON\u00a0=\u00a0104(72) kcal; HIGH\u00a0=\u00a0104(54) kcal; VHIGH\u00a0=\u00a0126(46) kcal; p\u00a0=\u00a00.220, \u03b72\u00a0=\u00a00.155) nor the total energy from carbohydrates (CON\u00a0=\u00a0152(39) kcal; HIGH\u00a0=\u00a0130(46) kcal; VHIGH\u00a0=\u00a0121(30) kcal; p\u00a0=\u00a00.113, \u03b72\u00a0=\u00a00.215) differed between conditions. Similarly, the average RER during the OGTT did not differ between conditions (CON\u00a0=\u00a00.90(0.08); HIGH\u00a0=\u00a00.87(0.06); VHIGH\u00a0=\u00a00.85(0.04); p\u00a0=\u00a00.092, \u03b72\u00a0=\u00a00.233). Hunger score was unaffected by the hypoxic intervention (CON\u00a0=\u00a0220(155) A.U; HIGH\u00a0=\u00a0253(146) A.U; VHIGH\u00a0=\u00a0240(123) A.U; p\u00a0=\u00a00.331, \u03b72\u00a0=\u00a00.116).Total energy expenditure during the OGTT differed between conditions (CON\u00a0=\u00a0256(39) kcal; HIGH\u00a0=\u00a0235(38) kcal; VHIGH\u00a0=\u00a0247(32) kcal; \u22121; HIGH\u00a0=\u00a064(6) b\u2219min\u22121; VHIGH\u00a0=\u00a073(10) b\u2219min\u22121; p\u00a0=\u00a00.008, \u03b72\u00a0=\u00a00.629 (n\u00a0=\u00a08)), being higher in both HIGH and VHIGH than CON (both p\u2009<\u20090.01), and higher in VHIGH than HIGH (p\u2009<\u20090.05), but neither systolic blood pressure (CON\u00a0=\u00a0129(10) mmHg; HIGH\u00a0=\u00a0122(9) mmHg; VHIGH\u00a0=\u00a0124(9) mmHg; p\u00a0=\u00a00.194, \u03b72\u00a0=\u00a00.167), diastolic blood pressure (CON\u00a0=\u00a080(8) mmHg; HIGH\u00a0=\u00a077(6) mmHg; VHIGH\u00a0=\u00a078(8) mmHg; p\u00a0=\u00a00.574, \u03b72\u00a0=\u00a00.060), nor mean arterial pressure (CON\u00a0=\u00a096(8) mmHg; HIGH\u00a0=\u00a092(7) mmHg; VHIGH\u00a0=\u00a093(6) mmHg; p\u00a0=\u00a00.263, \u03b72\u00a0=\u00a00.138) differed.Heart rate over the normobaric\u2010hypoxic exposure period differed between conditions (CON\u00a0=\u00a061(5) b\u2219minp\u00a0=\u00a00.009), which were greater in VHIGH compared to HIGH and CON (p\u2009<\u20090.05), with two participants meeting the criteria for AMS , being higher in VHIGH than HIGH or CON (p\u2009<\u20090.05).There was a between\u2010conditions difference in AMS symptoms following the normobaric\u2010hypoxic exposures (CON\u00a0=\u00a00(0\u20132); HIGH\u00a0=\u00a00(0\u20131); VHIGH\u00a0=\u00a01(0\u20137); 4IO2 (VHIGH).The aim of the present study was to assess the short\u2010term effects of different levels of acute hypoxia on subsequent glucose tolerance, insulin sensitivity, markers of inflammation and oxidative stress, and feasibility in males with overweight. Our main findings were that, following 60\u2009min of breathing either of two hypoxic gas mixtures: (1) neither glucose tolerance, as assessed by peak [glucose] and [glucose] AUC, insulin sensitivity, nor metabolic clearance rate of glucose significantly differed across conditions; (2) markers of oxidative stress and inflammation were not elevated by the acute hypoxic exposures; and (3) feasibility was reduced at the lowest FIO2 of 0.148 did not differ from a normoxic control, and Morishima and Goto\u00a0(IO2\u00a0=\u00a00.155) on resting postprandial glucose regulation. They differ, however, from those showing beneficial effects of acute hypoxia on subsequent glycaemic control in metabolically compromised individuals indicates that detection of a between\u2010conditions difference would have been possible with 17 participants .Hypoxia has been shown to increase muscle glucose uptake through effects on insulin\u2010independent the highest level of hypoxia that is reliably tolerated without adverse symptoms in individuals with overweight, (ii) the minimal effective exposure duration to this level of hypoxia in this cohort, and (iii) the recruitment of individuals with overweight with overtly compromised glycaemic control.Written informed consent was obtained before participation. The study was approved by the University's Faculty of Science and Health, Research Ethics Committee and conformed to the Declaration of Helsinki, except for registration in a database."} +{"text": "II\u2013di\u00adthio\u00adether complex. It crystallizes in the monoclinic space group P21/c. Additional Hirshfeld analyses indicate a C\u2014H\u22ef\u03c0 inter\u00adaction along the [010] axis to be the most important packing factor.The title complex represents a further example of a square-planar Pt 2(C7H8I2)2], represents a further example of a square-planar PtII\u2013di\u00adthio\u00adether complex. It crystallizes in the monoclinic space group P21/c. Additional Hirshfeld analyses indicate a C\u2014H\u22ef\u03c0 inter\u00adaction along the [010] axis to be the most important packing factor.The title complex, [PtI Alternatively, this air-stable complex could be prepared in a much improved yield of 80% by reaction of bis\u00ad(benzo\u00adnitrile)\u00addiiodo\u00adplatinum with 2 equivalents of methyl phenyl sulfide (thio\u00adanisol) MeSPh using di\u00adchloro\u00admethane as solvent. This compound was characterized by NMR spectroscopy in solution and exhibits a singlet resonance for the two magnetically equivalent methyl groups at \u03b4 3.01 ppm, flanked by 195Pt satellites due to a 3JPtH coupling of 48\u2005Hz. Furthermore, we report herein on the solid-state structure and structural analysis of trans-di\u00adiodido\u00adbis\u00ad[(methyl\u00adsulfan\u00adyl)benzene-\u03baS]platinum(II) (1). In addition, the results of a Hirshfeld analysis of the inter\u00admolecular inter\u00adactions are presented.In the past, our groups have investigated the coordination of chelating di\u00adthio\u00adethers such as the vinylic ferrocenyl-di\u00adthio\u00adether 2.trans-Di\u00adiodido\u00adbis\u00ad[(methyl\u00adsulfan\u00adyl)benzene-\u03baS]platinum(II) (1) crystallizes from di\u00adchloro\u00admethane in the monoclinic crystal system, space group P21/c. The mol\u00adecular structure of 1 is presented in Figs.\u00a021, which shows Ch2 symmetry. The distance from the coordinating iodine center I1 to Pt1 is 2.61205\u2005(15)\u2005\u00c5, showing a slight elongation with respect to its educt structure trans-[PtI2(NCPh)2] (2) (2.6052\u2005(8)\u2005\u00c5; Viola et al., 2018trans-[PtCl2(SMePh)2] (3) reported by Ahlgr\u00e9n 2] (4) 2] (5) \u2005\u00c5 in 5]. The chelate complexes cis-di\u00adiodo-\u00adplatinum(II) di\u00adiodoplatinum(II) \u00b0, C2\u2014S1\u2014Pt1 = 104.52\u2005(7)\u00b0 and C2\u2014S1\u2014C1 = 103.46\u2005(11)\u00b0.All further bonds have characteristic dimensions x, y, \u2212z] with a distance between the phenyl ring and H1Bi of 2.5377\u2005(10)\u2005\u00c5 benzene] ligand and its oxidized derivative are focused on now. The already compared structure trans-di\u00adchloro-bis\u00ad[meth\u00adyl(phen\u00adyl)sulfan\u00adyl]platinum \u00adpalladium has been published independently by two different research groups [JISWUD platinum(II) platinum(II) 2SiPh2}] 2SiPh2}]\u00b7DCM 2Si2Me4}] 2Si2Me4}]\u00b7DCM benzene]\u00adplatinum (1) was synthesized by adding methyl\u00adphenyl sulfide dissolved in 0.5\u2005mL of di\u00adchloro\u00admethane via a microsyringe to a solution of bis\u00ad(benzo\u00adnitrile)\u00addiiodo\u00adplatinum in di\u00adchloro\u00admethane (3\u2005mL) and stirring overnight at room temperature. trans-Di\u00adiodo\u00adbis\u00ad[(methyl\u00adsulfan\u00adyl)benzene]\u00adplatinum was isolated as red crystals after layering with heptane.14H16I2PtS2 (697.30\u2005g\u2005mol\u22121): C, 24.11; H, 2.32; S, 9.20. Found: C, 23.92; H, 2.21; S, 9.05%.Calculated for C1H NMR : \u03b4 = 3.01 , 7.05\u20137.73 ppm.6.Uiso(H) = 1.2Ueq(C) for CH2 and CH hydrogen atoms and Uiso (H) = 1.5Ueq(C) for CH3 hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023003717/jy2030sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989023003717/jy2030Isup2.hklStructure factors: contains datablock(s) I. DOI: 2258409CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The salts were characterized by the multinuclear NMR and IR spectroscopy as well as X\u2010ray diffraction. Hirshfeld surface analysis and solid state structures reveal various intermolecular anion\u2010\u03c0 and \u03c3\u2010hole interactions between the corresponding halogenated pyridinium cations and the anion [Al(OTeF5)4]\u2212.The synthesis and the first structural characterization of the halogenated pyridinium salts [C 5F5NH]+ and [C5F4ClNH]+ and the dimeric cations [(C5F5N)2H]+ and [(C5Cl5N)2H]+ as [Al(OTeF5)4]\u2212 salts are presented. These cations show noncovalent interactions with the weakly coordinating anion [Al(OTeF5)4]\u2212 such as strong fluorine specific interactions as well as \u03c3\u2010hole and anion\u2010\u03c0 interactions.The synthesis and structural characterization of the perhalogenated pyridinium cations [C While non fluorinated pyridine can be protonated by HCl or HBr, pentafluoropyridine can only be protonated by Br\u00f8nsted superacids such as HF/AsF5 and HF/SbF5.o\u2010C6H4F2\u2010H][Al(OTeF5)4] is known for the protonation of weak bases like benzene and white phosphorus, resulting in [C6H7]+ and [P4H]+, respectively.6F6, which can result in anion\u2010\u03c0 interactions.4]\u2212 and [PF6]\u2212[6] or between perfluoroarenes, like C5F5N, C6F6 and C10F8 with different halide anions.5F5N, C5F4ClN, and C5Cl5N by [o\u2010C6H4F2\u2010H][Al(OTeF5)4] (1\u2009a) and studied hydrogen bonding, halogen bonding and anion\u2010\u03c0 interactions between pentafluororopyridinium [C5F5NH]+ (2), 4\u2010chloro\u20102,3,5,6\u2010tetrafluoropyridinium [C5F4ClNH]+ (3), pentachlororopyridinium [C5Cl5NH]+ (4) and the fluorinated WCA [Al(OTeF5)4]\u2212 (a).In contrast to pyridine, the perfluorinated pyridine C5F5N2\u03c3(I)) and wR2 was 0.0484 .The authors declare no conflict of interest.1As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "The crystal structures and Hirshfeld surface analyses of three salts of 1-(4-nitro\u00adphenyl)\u00adpiperazine with 2-chloro\u00adbenzoic acid, 2-bromo\u00adbenzoic acid and 2-iodo\u00adbenzoic acid are reported. 10H14N3O2+\u00b7C7H4ClO2\u2212, contains whole-ion-disordered cations and anions, which were modeled with two equivalent conformations with occupancies of 0.745\u2005(10)/0.255\u2005(10) and 0.563\u2005(13)/0.437\u2005(13), respectively. The bromo\u00adbenzoate and iodo\u00adbenzoate derivatives are isomorphous and crystallize as hemihydrates, viz. C10H14N3O2+\u00b7C7H4BrO2\u2212\u00b70.5H2O and C10H14N3O2+\u00b7C7H4IO2\u2212\u00b70.5H2O, respectively [the water mol\u00adecule is disordered over two locations with occupancies of 0.276\u2005(3)/0.223\u2005(3) for the iodo\u00adbenzoate derivative]. In the extended structures, all three salts feature an R44(12) loop of two anions and two cations linked by N\u2014H\u22efO hydrogen bonds.The crystal structures and Hirshfeld surface analyses of three salts of 1-(4-nitro\u00adphenyl)\u00adpiperazine with 2-chloro\u00adbenzoic acid, 2-bromo\u00adbenzoic acid and 2-iodo\u00adbenzoic acid are reported. The chloro\u00adbenzoate salt, C When discussing the conformations of the anion and cation, only the major components will be used. In the chloro\u00adbenzoate anion, the carboxyl\u00adate group is significantly twisted with respect to the 2-chloro\u00adphenyl ring with a dihedral angle of 76.7\u2005(4)\u00b0, which is likely due to the steric inter\u00adaction between the ortho-chloro substituent and the carboxyl\u00adate group. Structures 2 and 3 exhibit similar cation conformations, with equivalent dihedral angles of 65.5\u2005(3) and 67.1\u2005(5)\u00b0, respectively. Additionally, in all three structures, the 4-nitro\u00adphenyl group occupies an equatorial position in its attachment to the piperazinium ring.Structure ds Fig.\u00a01, which w2 and 3 are isostructural, only 2 will be discussed in detail. This structure consists of a 4-(4-nitrophenyl)piperazin-1-ium cation linked to a 2-bromo\u00adbenzoate anion by two N\u2014H\u22efO hydrogen bonds /0.223\u2005(3) for the iodo\u00adbenzoate derivative]. Additionally, there is a weak C\u2014H\u22efBr inter\u00adaction accepted by the bromine atom in the 2-bromo\u00adbezoate anion and a carbon atom in the piperazinium ring, as well as a pair of weak C\u2014H\u22efO inter\u00adactions between adjacent 4-nitro\u00adphenyl rings in the 4-(4-nitrophenyl)piperazin-1-ium cation.Since s Figs. 2 and 3 \u25b8.3.1, which contains both a disordered cation and anion as well as disordered water of solvation, the discussion will focus solely on the major component. The cation forms an x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z; x, 1\u00a0+\u00a0y, z; 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z; see Fig.\u00a04x, \u2212y, \u2212z; Y\u22efCg distance = 3.488\u2005(18)\u2005\u00c5; X\u2014Y\u22efCg: 85.8\u2005(12)\u00b0].In the packing of 2, two cations and two anions form an et al., 1990x, \u2212y, \u2212z; see Fig.\u00a06x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z) that form an x, y, z). The phenyl rings in adjacent cations form \u03c0\u2013\u03c0 inter\u00adactions with a perpendicular distance between centroids of 3.5332\u2005(11)\u2005\u00c5 . These are all clearly seen in the fingerprint plot generated by CrystalExplorer that form an Refinement). This species is likely to be involved in hydrogen bonding with an adjacent oxygen atom in the anion and with the piperazine ring in the cation, forming an x, 1\u00a0\u2212\u00a0y, \u2212z; slippage = 0.379\u2005\u00c5).In the packing of 4.et al., 2014et al., 2022aet al., 2022et al., 2022aRelated structures containing the 4-(4-nitrophenyl)piperazin-1-ium cation include 4-(4-nitrophenyl)piperazin-1-ium chloride monohydrate \u2013(3), a solution of commercially available (from Sigma-Aldrich) 1-(4-nitro\u00adphenyl)\u00adpiperazine in methanol (10\u2005ml) was mixed with equimolar solutions of the appropriate acids in methanol (10\u2005ml) and ethyl acetate (10\u2005ml), viz., 2-chloro\u00adbenzoic acid for (1), 2-bromo\u00adbenzoic acid for (2), and 2-iodo\u00adbenzoic acid for (3), The resulting solutions were stirred for 15 minutes at room temperature and allowed to stand at the same temperature. X-ray quality crystals were formed on slow evaporation after one week for all compounds, where ethanol:ethyl\u00adacetate (1:1) was used for crystallization. The melting points are 439\u2013441\u2005K (1), 443\u2013445\u2005K (2) and 451\u2013453\u2005K (3).For the synthesis of salts (6.Uiso(H) = 1.2Ueq(C) while the N\u2014H hydrogen atoms were refined isotropically. For 1, in which both the cation and the anion exhibit whole-ion disorder, two equivalent conformations were modeled with occupancies of 0.745\u2005(10)/0.255\u2005(10) and 0.563\u2005(13)/ 0.437\u2005(13) respectively. The water hydrogen atoms were refined isotropically with idealized geometries.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S205698902300302X/hb8051sup1.cifCrystal structure: contains datablock(s) 1, 2, 3. DOI: 10.1107/S205698902300302X/hb80511sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S205698902300302X/hb80512sup3.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S205698902300302X/hb80513sup4.hklStructure factors: contains datablock(s) 3. DOI: 2253382, 2253381, 2253380CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "After the introduction of a pyridine\u2010containing auxiliary, which provided access to challenging junctions (proline or \u03b2\u2010branched amino acids), we herein probe the role of the pyridine\u2010ring nitrogen. We observed side reactions leading to preliminary auxiliary loss. We describe a new easy to attach \u03b2\u2010mercapto\u2010\u03b2\u2010(4\u2010methoxy\u20102\u2010pyridinyl)\u2010ethyl (MMPyE) auxiliary, which 1) has increased stability; 2) enables NCL at sterically encumbered junctions ; and 3) allows removal under mildly basic (pH\u20058.5) conditions was introduced. The synthesis of a 120 aa long peptide containing eight MUC5AC tandem repeats via ligation of two 60mers demonstrates the usefulness. Making use of hitherto unexplored NCL to tyrosine, the MMPyE auxiliary provided access to a head\u2010to\u2010tail\u2010cyclized 21\u2010mer peptide and a His A properly positioned pyridine ring can speed up auxiliary\u2010mediated native chemical ligation. A methoxy substituent increases auxiliary stability and facilitates rapid syntheses of long and multiphosphorylated multirepeat proteins and of head\u2010to\u2010tail cyclized peptides. N\u2010terminal cysteine for reactions with peptide thioesters. The ligation\u2010desulfurization involves thiolated amino acids, which mimic the 1,2\u2010aminothiol structure of cysteine. After ligation, the thiol group is removed by a radical reaction.The native chemical ligation (NCL) reaction is a cornerstone of protein total synthesis.N\u2010terminus.N\u2010terminal alkylation reduces the reactivity of the amino group. Exemplary for this problem are the early N\u2010benzyl type auxiliaries will become challenging if not impossible at amino acids with larger steric demand than glycine. A solution to the problem became apparent when we, while working on alternative auxiliary scaffolds, became aware of a radical fragmentation reaction (see below) that allows removal of non\u2010benzyl auxiliaries.8 ethyl (MPyE) scaffold 9 is the first auxiliary enabling ligations at sterically demanding junctions such as Leu\u2010Arg, Leu\u2010Val or Pro\u2010Ala.S,N\u2010acyl transfer by base catalysis . While shifting the nitrogen from the ortho to the para position should have little effect on its basicity, the intramolecular proton abstraction should be impossible. For the para isomer of the MPyE auxiliary, we therefore expected a ligation speed comparable to that of the MPE auxiliary 8.The recently introduced MPyE auxiliary enables NCL reactions at ligation junctions inaccessible to previous ligation auxiliaries. For example, we established Ala\u2010Val, Leu\u2010Val, Pro\u2010Ala and Pro\u2010Arg bonds.17 . Under these conditions, the original ortho isomer of the MPyE auxiliary remained stable. Nevertheless, with quick and careful handling we succeeded in the isolation of product (53\u2009%) and subsequently analyzed reactions of the para\u2010MPyE peptide with the peptide thioester 19A. The attempted Ala\u2010Asn ligation was slow . The decrease of ligation rates upon para positioning illustrates the role of the ring nitrogen, which must be in the ortho position to allow acceleration of the S,N\u2010acyl transfer by intramolecular base catalysis.The aldehyde precursor 7 Figure\u2005A requirepara\u2010MPyE group from peptide 18N prompted us to also critically analyze the stability of the ortho\u2010MPyE auxiliary. In a published example23LV and iminium ion 31. The latter would undergo rapid hydrolysis. Alternatively, the ethylpyridine structures may undergo a Hoffman elimination facilitated by anchimeric assistance from a deprotonated thiol group, which would be considerably acidified by pyridine protonation =4.88; pKA (pyridine)=5.17)The considerations on the stability and reactivity of the MPyE auxiliary indicate an important role of the ring nitrogen. We examined methoxy substitution in meta position to the nitrogen. Methoxy substitution decreases the basicity of pyridine nitrogen (pK\u03b1\u2010methyl carbon with lithium diisopropylamide and subjected to a nucleophilic acylation with diethyl carbonate to yield methyl (5\u2010methoxy\u20102\u2010pyridinyl)acetate (37). Treatment with lithium bis(trimethylsilyl)amide afforded an ester enolate which was converted to the \u03b1\u2010mercapto ester 39 in an electrophilic thiolation. Reduction of 39 with diisobutylaluminium hydride (DIBALH) furnished 40 as a suitable precursor for auxiliary introduction by reductive alkylation. Compound 40 is present in its enole form and shelf\u2010stable.For the introduction of the MMPyE auxiliary, a suitable precursor was synthesized in 4 steps in presence of 2\u2009% acetic acid, the peptides were treated with a mixture of TFA/H2O/TIS (95/2.5/2.5 v/v/v) for final deprotection and cleavage of the peptides from resin. The crude peptides were obtained in high purity, indicative of efficient reductive alkylation .We next attached the MMPyE auxiliary to the unprotected N\u2010terminus of asparagine and valine test peptides Figure\u2005B. After 41X with the peptide thioesters 19Z the compounds were dissolved in a mixture prepared by adding 3\u2005vol.% thiophenol to a phosphate buffer containing 6\u2005M guanidinium hydrochloride with an apparent pH\u20057.5. The ligations proceeded smoothly. Compared to reactions at the non\u2010substituted MPyE peptide we observed the expected decrease in the reaction rate , we observed a rapid auxiliary cleavage within 3 to 6\u2005h .52 was prepared by microwave\u2010assisted solid phase peptide synthesis (MW\u2010SPPS) and used in crude form in the reaction with mercaptopropionate (MPA). The conversion to the MPA ester 53 proceeded smoothly . For an approach to a peptide thioester, we first synthesized the peptide hydrazide by microwave\u2010assisted solid\u2010phase synthesis. Dawson's acetoacetone method was applied to establish the peptide thioester 59 without detriment to the three phospho sites .59 and 57 was discontinued by treatment with hydrazine. Though our aim was to develop a method providing rapid access of product without isolation of intermediates, we also analyzed the ligation mixture . We observed an unexpected by\u2010product resulting from cleavage of the N\u2010terminal tyrosine from peptide 57. This side reaction was less pronounced in ligations with non\u2010phosphorylated peptides and did not occur at all with a test peptide . At current, we cannot explain this unusual side reaction. Other ligations at tyrosine 6 peptide 60 in pure form.Due to our interest in post\u2010translationally modified multirepeat proteins, we tested the auxiliary in the rapid synthesis of a hexaphosphorylated peptide comprising 6 heptapeptide repeat units from the C\u2010terminal domain of the large RNA polymerase II subunit.In a further exploration of application scenarios, we studied a cyclopeptide. The chemical synthesis of cyclopeptides is an active area in medicinal peptide chemistry and NCL is powerful method to achieve cyclization of unprotected linear peptide precursors.Previously we have shown that cysteine residues should be protected during auxiliary removal.\u03b2\u2010mercapto\u2010\u03b2\u2010(2\u2010pyridinyl)\u2010ethyl auxiliaries proceed in high rate and enable joining of peptide fragments at junctions, such as the Leu\u2010Val bond inaccessible to previous ligation auxiliaries. In this work, we probed the role of the ortho\u2010positioned ring nitrogen. A comparison with the para isomer (\u03b2\u2010mercapto\u2010\u03b2\u2010(4\u2010pyridinyl)\u2010ethyl) scaffold revealed the superior reactivity of the ortho pyridine derivatives. We attribute the high reactivity to the ortho ring nitrogen abstracting a proton from an ammonium ion formed when the amino group intramolecularly attacks the thioester bond. Without deprotonation the S,N\u2010acyl shift cannot proceed and the reaction stops at the thioester intermediate stage. Our investigations also uncovered a peculiar lability of peptides modified with the \u03b2\u2010mercapto\u2010\u03b2\u2010(4\u2010pyridinyl)\u2010ethyl) group at the N\u2010terminus. While the para\u2010pyridine auxiliary\u2010peptide bond was stable during TFA cleavage, rapid cleavage occurred at moderately acidic pH\u20052. This reaction only occurs prior to ligation indicating a role of the amino terminus\u2019 lone pair. Though at a slower rate, premature auxiliary loss was also observed with the ortho\u2010system (\u03b2\u2010mercapto\u2010\u03b2\u2010(2\u2010pyridinyl)\u2010ethyl). Importantly, our studies show that the auxiliary gains stability when a methoxy substituent is added at the 4\u2010position of the ortho\u2010pyridine ring (i.\u2009e. meta to the ring nitrogen).Native chemical ligation reactions on \u03b2\u2010mercapto\u2010\u03b2\u2010(4\u2010methoxy\u20102\u2010pyridinyl)\u2010ethyl) auxiliary is conveniently attached to the peptide N\u2010terminus in the last step of solid\u2010phase synthesis by reductive alkylation. Despite slightly lowered ligation rates, the MMPyE auxiliary is better suited for NCL at challenging junctions than the MPyE auxiliary becausethe increased stability is a significant advantage when reactions such as the selenoester\u2010NCLs are performed at slightly acidic conditions. Reactions proceed cleanly also when long reaction times are needed in ligations of large peptides. Furthermore, the increased stability enables ligations at elevated temperature. The usefulness of the MMPyE auxiliary was demonstrated in three applications. The synthesis of a 120 aa long peptide containing 8 MUC5AC tandem repeats and two \u201csticky\u201d coiled coil peptides involved the smooth ligation of two 60mers via a Ser\u2010Ala junction. Using a hitherto unexplored ligation site in NCL chemistry and demonstrating the potential for the preparation of post\u2010translationally modified peptides, we applied the MMPyE auxiliary in a Ser\u2010Tyr ligation for the synthesis of a His6\u2010tagged hexaphosphorylated peptide spanning 6 tandem repeats of the RNA polymerase II C\u2010terminal domain. The rapid synthesis of a Phe\u2010Tyr\u2010cyclized 21 aa peptides comprising the primary sequence of microcin J25 illustrates an application of the auxiliary in head\u2010to\u2010tail cyclization reactions.The new MMPyE are stable over a wide pH range, 2) enable native chemical ligations at sterically encumbered junctions, 3) allow removal under mildly basic (pH\u20058.5) conditions and 4) provide options for attaching further handles by means of 4\u2010alkoxy substituents. The last option offers prospects for the introduction of functional units such as oligoethylene glycol or oligoarginine units that enhance the solubility of unfolded peptide segments.Ethyl 2\u2010(pyridin\u20104\u2010yl)\u20102\u2010thio)acetate (S4): Under argon atmosphere, lithium hexamethyldisilazide was added to a stirred solution of ethyl 2\u2010(pyridin\u20104\u2010yl)acetate in 75\u2005mL anhydrous THF over the course of 25\u2005min at \u221278\u2009\u00b0C. After 30\u2005min of stirring, thiosulfonate 38 in 40\u2005mL anhydrous THF was added at \u221278\u2009\u00b0C over 100\u2005min. The reaction mixture was allowed to gradually warm to room temperature over the course of 30\u2005min and a saturated solution of NH4Cl was added. The resultant mixture was extracted with EtOAc (3x) and the combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (EtOAc) to afford desired product in 82\u2009% yield as a white solid. 1H NMR : \u03b4 [ppm]=8.52 , 7.38 , 6.16 , 4.66 , 4.12 , 3.78 , 3.75 , 3.75 \u2010 3.72 , 1.19 . 13C NMR : \u03b4 [ppm]=170.94, 161.94, 159.85, 150.77, 147.17, 124.38, 107.02, 91.64, 62.74, 56.44, 56.06, 52.28, 25.19, 14.32.2\u2010(Pyridin\u20104\u2010yl)\u20102\u2010thio)acetaldehyde (17): Under argon atmosphere ester S4 was dissolved in 5\u2005mL anhydrous DCM and cooled to \u201094\u2009\u00b0C. Diisobutylaluminium hydride was added carefully over the vessel wall over the course of 1\u2005h. The reaction mixture was stirred for an additional 15\u2005min at \u221294\u2009\u00b0C. Subsequently, a mixture of DCM/MeOH was added via the vessel wall over the course of 30\u2005min at \u221294\u2009\u00b0C. The reaction mixture was stirred at room temperature for 10\u2005min and a saturated solution of potassium sodium tartrate and DCM (20\u2005ml each) was added. The resulting mixture was stirred at room temperature for 1\u2005h and then extracted with DCM (3x). The combined organic layers were dried over MgSO4, filtered and concentrated. The residue was purified by flash column chromatography on silica gel (EtOAc) to afford the aldehyde 17 (247\u2005mg) as yellow oil. The material was used for reductive alkylation despite impurities from remaining solvents and partial decomposition. 1H NMR : \u03b4 [ppm]=9.52 , 8.52 , 7.27 , 6.10 , 4.62 , 3.78 , 3.77 , 3.75 .Ethyl 2\u2010(5\u2010methoxypyridin\u20102\u2010yl)acetate (37): nBuLi was added dropwise to a stirred solution of diisopropylamine in 20\u2005ml anhydrous THF at \u221278\u2009\u00b0C under argon. The resulting solution was slowly warmed to room temperature and was added dropwise over 1\u2005h to a stirred solution of 5\u2010methoxy\u20102\u2010methylpyridine and anhydrous diethylcarbonate in 26\u2005ml anhydrous THF at \u221278\u2009\u00b0C. After 2\u2005h of reaction, the solution was slowly warmed to room temperature and a saturated solution of NH4Cl was added. The resultant mixture was extracted with EtOAc (3x), and the combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (cyclohexane/EtOAc=6\u2009:\u20094 v/v) to afford 37 as a light\u2010yellow oil. 1H NMR : \u03b4 [ppm]=8.25 , 7.28\u20137.20 , 4.18 , 3.86 , 3.82 , 1.26 . 13C NMR : \u03b4 [ppm]=170.91, 154.88, 146.23, 136.13, 124.52, 122.22, 61.17, 55.83, 42.59, 14.28. HRMS: m/z=196.0967 (C10H14NO3 (M+H)+, calcd.: 196.0968).Ethyl 2\u2010(5\u2010methoxypyridin\u20102\u2010yl)\u20102\u2010thio) acetate (39): Under argon atmosphere, lithium hexamethyldisilazide was added to a stirred solution of ethyl 2\u2010(5\u2010methoxypyridin\u20102\u2010yl)acetate in 40\u2005mL anhydrous THF over the course of 25\u2005min at \u221278\u2009\u00b0C. After 30\u2005min of stirring, thiosulfonate 38 in 37\u2005mL anhydrous THF was added at \u221278\u2009\u00b0C within 100\u2005min. The reaction mixture was allowed to gradually warm to room temperature over the course of 30\u2005min and a saturated solution of NH4Cl was added. The resultant mixture was extracted with EtOAc (3x) and the combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (cyclohexane/EtOAc=1\u2009:\u20091 v/v) to afford 39 as a white solid. 1H NMR : \u03b4 [ppm]=8.15 , 7.43 , 7.27 , 6.17 , 4.75 , 4.12 , 3.83 , 3.78 , 3.76 , 1.19 . 13C NMR : \u03b4 [ppm]=171.39, 161.69, 159.74, 155.98, 149.63, 137.36, 124.05, 122.02, 107.32, 91.52, 62.23, 56.37, 56.33, 55.95, 54.59, 25.03, 14.33. HRMS: m/z=408.1455 (C20H26NO6S (M+H)+, calcd.: 408.1475).E)\u20102\u2010(5\u2010methoxypyridin\u20102\u2010yl)\u20102\u2010thio)ethenol (40) was dissolved in 2.1\u2005mL anhydrous DCM and cooled to \u201094\u2009\u00b0C. Diisobutylaluminium hydride was added carefully via the vessel wall over the course of 1\u2005h. The reaction mixture was stirred for an additional 15\u2005min at \u221294\u2009\u00b0C. Subsequently, a mixture of DCM/MeOH was added via the vessel wall over the course of 30\u2005min at \u221294\u2009\u00b0C. The reaction mixture was stirred at room temperature for 10\u2005min and a saturated solution of potassium sodium tartrate and DCM (20\u2005ml each) was added. The resulting mixture was stirred at room temperature for 1\u2005h and then extracted with DCM (3x). The combined organic layers were dried over MgSO4, filtered and concentrated. The residue was purified by recrystallization (MeOH/H2O) to afford the enol 40 as bright yellow crystals. 1H NMR : \u03b4 [ppm]=14.83 , 8.07 , 7.79 , 7.45 , 7.15 , 6.11 , 3.86 , 3.76 , 3.70 , 3.61 . 13C NMR : \u03b4 [ppm]=162.21, 161.52, 159.95, 154.14, 153.60, 132.01, 124.77, 122.72, 107.40, 102.62, 91.34, 56.68, 56.29, 55.97, 28.49. HRMS: m/z=364.1201 (C18H22NO5S (M+H)+, calcd.: 364.1213).Peptide Purification: The desired peptides were purified by preparative HPLC on a C18\u2010column with a binary mixture of A and B as mobile phase (flow=6.0\u2005mL/min). For model peptides a linear gradient from 3\u201330\u2009% B was used. For other peptides a linear gradient was used as indicated. All products were isolated as white solids after lyophilization. Yields were determined by measurements of UV absorption as described in chapter 5.3. of the Suppl. Inf. or by weighing.2)2(CO)\u2010Gly\u2010NH2 (19A)LYRAA\u2010S(CH: Thioester 19A was synthesized according to the published procedure.R=1.39\u2005min (3\u201330\u2009% B in 4\u2005min); m/z=737.5 (C32H53N10O8S (M+H)+, calcd.: 737.4), 369.3 (C32H54N10O8S (M+2H)2+, calcd.: 369.2); C32H52N10O8S (MW=736.4\u2005g\u2009mol\u22121).2)2(CO)\u2010Gly\u2010NH2 (19L)LYRAL\u2010S(CH: Thioester 19L was synthesized according to the published procedure.R=3.57\u2005min (3\u201335\u2009% B in 6\u2005min); m/z=779.7 (C35H59N10O8S (M+H)+, calcd.: 779.4), 390.4 (C35H60N10O8S (M+2H)2+, calcd.: 390.2); C35H58N10O8S (MW=778.4\u2005g\u2009mol\u22121).LYRAA\u2010SePh (22A): Selenoester 22A was synthesized according to the published procedure.R=5.31\u2005min (3\u201330\u2009% B in 6\u2005min); m/z=733.4 (C33H49N8O6Se (M+H)+, calcd.: 733.3), 367.4 (C33H50N8O6Se (M+2H)2+, calcd.: 367.1); C33H48N8O6Se (MW=732.3\u2005g\u2009mol\u22121).LYRAL\u2010SePh (22L): Selenoester 22L was synthesized according to the published procedure.R=2.31\u2005min (3\u201060\u2009% B in 4\u2005min); m/z=775.4 (C36H55N8O6Se (M+H)+, calcd.: 775.3), 388.3 (C36H56N8O6Se (M+2H)2+, calcd.: 388.2); C36H54N8O6Se (MW=774.3\u2005g\u2009mol\u22121).General Procedure for Introduction of MMPyE Auxiliary: The peptidyl\u2010resin was allowed to swell in MeOH/NMP/AcOH (6\u2009:\u20094\u2009:\u20090.02 v/v/v) for 15\u2005min and then treated with a mixture of the enol 40 and NaCNBH3 in MeOH/NMP/AcOH (3\u2009:\u20091\u2009:\u20090.02 v/v/v) over night at room temperature . Afterwards the resin was washed with DCM (5x), MeOH (5x) and DCM (5x) and dried under vacuum. Finally, the peptide was deprotected and cleaved of the resin by addition of a mixture of TFA\u2009:\u2009TIS\u2009:\u2009H2O . After 18\u201324\u2005h the cleavage cocktail was collected by filtration and the resin was washed with TFA (3\u00d70.5\u2005mL). The combined filtrates were concentrated and ice cold Et2O (\u223c8\u201310\u2010fold volume) was added. The suspension was centrifuged and the ether phase was decanted. For HPLC purification, the peptide pellet was dissolved in H2O\u2009:\u2009ACN\u2009:\u2009TFA (1\u2009:\u20091\u2009:\u20090.001 v/v/v).pMPyE\u2010NRAEYSGLG (18N): Synthesis of 16N was achieved by microwave\u2010assisted solid phase peptide synthesis . The auxiliary was introduced according to the published procedure for the MPyE auxiliary2O/TIS (95\u2009:\u20092.5\u2009:\u20092.5 v/v/v) for 18\u2005h. After analyzing the resulting fractions with UPLC\u2010MS, the auxiliary peptide containing solutions were swiftly frozen for lyophilization to avoid decomposition of the product. Yield: 7.69\u2005mg, 53\u2009%. Note: 18N was stored as dry lyophilizate at \u221280\u2009\u00b0C to avoid auxiliary cleavage . UPLC\u2010MS: tR=2.87\u2005min (3\u201330\u2009% B in 6\u2005min); m/z=1102.6 (C47H72N15O14S (M+H)+, calcd.: 1102.5), 552.1 (C47H73N15O14S (M+2H)2+, calcd.: 551.8); C47H71N15O14S (MW=1102.2\u2005g\u2009mol\u22121).MMPyE\u2010NRAEYSGLG (41N): Yield: 6.55\u2005\u03bcmol, 66\u2009%. UPLC\u2010MS: tR=3.71\u2005min (3\u201330\u2009% B in 6\u2005min); m/z=1132.6 (C48H74N15O15S (M+H)+, calcd.: 1132.5), 566.9 (C48H75N15O15S (M+2H)2+, calcd.: 566.8); C48H73N15O15S (MW=1132.3\u2005g\u2009mol\u22121).MMPyE\u2010VRAEYSGLG (41V): Yield: 6.15\u2005\u03bcmol, 41\u2009%. UPLC\u2010MS: tR=4.33\u2005min (3\u201330\u2009% B in 6\u2005min); m/z=1117.7 (C49H77N14O14S (M+H)+, calcd.: 1117.6), 559.4 (C49H78N14O14S (M+2H)2+, calcd.: 559.3); C48H74N14O13S (MW=1117.3\u2005g\u2009mol\u22121).Ligation of Model Peptides with Peptide Thioester: To the ligation buffer 3\u2005vol.% thiophenol was added and the mixture was transferred to a lyophilized mixture of auxiliary peptide (1\u2005equiv) and peptide thioester (1.5\u2005equiv) to a final concentration of 5\u2005mM auxiliary peptide. An argon atmosphere was applied to the reaction vessel. To monitor the progress of the reaction, aliquots were withdrawn from the ligation mixture, quenched with an aqueous solution of 0.1\u2009% TFA or 0.1\u2009% TFA, 2.5\u2009% hydrazine, 30\u2005mM TCEP and analyzed by UPLC\u2010MS. The progress of the ligation reaction was assessed by integration of the corresponding peak areas. After completion of the ligation, hydrazine and TCEP were added to the ligation solution. Ligated peptides showed partial rearrangement back to the thioester intermediate via N\u2192S acyl transfer due to the acidic conditions used during and shortly after purification. Swift lyophilization after purification minimizes rearrangement. Alternatively, peptide solutions containing high amounts of rearranged thioesters can be treated with a solution of NH4Ac to a final pH of 5 to retrieve the amide.Ala\u2010Asn Ligation on pMPyE (20AN): 2\u2005\u03bcmol of para\u2010MPyE peptide 18N and 3\u2005\u03bcmol of peptide thioester 19A were allowed to react. The mixture was analyzed via UPLC\u2010MS after t=5\u2005min and t=24\u2005h : 2\u2005\u03bcmol of MMPyE\u2010peptide 41N and 3\u2005\u03bcmol of peptide thioester 19A were allowed to react for 72\u2005h. Yield: 1.02\u2005\u03bcmol, 51\u2009%. UPLC\u2010MS: tR=4.04\u2005min (3\u201330\u2009% B in 6\u2005min); m/z=854.0 (C75H118N24O20S (M+2H)2+, calcd.: 853.4), 569.9 (C75H118N24O20S (M+3H)3+, calcd.: 569.3); C75H116N24O20S (MW=1705.9\u2005g\u2009mol\u22121).Leu\u2010Asn Ligation on MMPyE (42LN): 2\u2005\u03bcmol of MMPyE\u2010peptide 41N and 3\u2005\u03bcmol of peptide thioester 19L were allowed to react for 72\u2005h. Yield: 1.14\u2005\u03bcmol, 57\u2009%. UPLC\u2010MS: tR = 5.00\u2005min (3\u201330\u2009% B in 6\u2005min); m/z=875.0 (C78H124N24O20S (M+2H)2+, calcd.: 874.5), 583.9 (C78H125N24O20S (M+3H)3+, calcd.: 583.3); C78H122N24O20S (MW=1748.0\u2005g\u22c5mol\u22121).Ala\u2010Val Ligation on MMPyE (42AV): 2\u2005\u03bcmol of MMPyE\u2010peptide 41V and 3\u2005\u03bcmol of peptide thioester 19A were allowed to react for 72\u2005h. Yield: 1.20\u2005\u03bcmol, 60\u2009%. UPLC\u2010MS: tR=6.07\u2005min (3\u201330\u2009% B in 8\u2005min); m/z=846.7 (C76H121N23O19S (M+2H)2+, calcd.: 845.9), 565.1 (C76H122N23O19S (M+3H)3+, calcd.: 564.3); C76H119N23O19S (MW=1691.0\u2005g\u2009mol\u22121).Leu\u2010Val Ligation on MMPyE (42LV): 2\u2005\u03bcmol of MMPyE\u2010peptide 41V and 3\u2005\u03bcmol of peptide thioester 19L were allowed to react for 120\u2005h. Yield: 0.58\u2005\u03bcmol, 29\u2009%. UPLC\u2010MS: tR=5.86\u2005min (3\u201335\u2009% B in 6\u2005min); m/z=867.5 (C76H127N23O19S (M+2H)2+, calcd.: 866.9), 578.9 (C76H128N23O19S (M+3H)3+, calcd.: 578.3); C76H125N23O19S (MW=1733.0\u2005g\u2009mol\u22121).Ligation with Model Peptide Selenoester: A lyophilized mixture of MMPyE peptide (1\u2005equiv) and peptide selenoester (2\u2005equiv) was dissolved in the ligation buffer , pH=6.2) to a final concentration of 5\u2005mM auxiliary peptide. An argon atmosphere was applied to the reaction vessel. To monitor the progress of the reaction, aliquots were withdrawn from the ligation mixture, quenched with an aqueous solution of 0.1\u2009% TFA or 0.1\u2009% TFA, 2.5\u2009% hydrazine, 30\u2005mM TCEP and analyzed by UPLC\u2010MS. The progress of the ligation reaction was assessed by integration of the corresponding peak areas. After completion of the ligation, hydrazine and TCEP were added to the ligation solution.Ala\u2010Asn Ligation on MMPyE (\u219242AN) with Selenoester 22A: 1\u2005\u03bcmol of MMPyE\u2010peptide 41N and 2\u2005\u03bcmol of peptide selenoester 22A allowed to react for 24\u2005h. Yield: 0.73\u2005\u03bcmol, 73\u2009%. UPLC\u2010MS: tR=4.05\u2005min (3\u201330\u2009% B in 6\u2005min); m/z=854.4 (C75H118N24O20S (M+2H)2+, calcd.: 853.4), 570.1 (C75H119N24O20S (M+3H)3+, calcd.: 569.3); C75H116N24O20S (MW=1705.9\u2005g\u2009mol\u22121).Leu\u2010Val Ligation on MMPyE (\u219242LV) with Selenoester 22A: 0.5\u2005\u03bcmol of MMPyE\u2010peptide 41V and 1.0\u2005\u03bcmol of peptide selenoester 22L were allowed to react for 72\u2005h. Yield: 0.24\u2005\u03bcmol, 48\u2009%. UPLC\u2010MS: tR=5.01\u2005min (3\u201340\u2009% B in 6\u2005min); m/z=867.9 (C76H127N23O19S (M+2H)2+, calcd.: 866.9), 579.1 (C76H128N23O19S (M+3H)3+, calcd.: 578.3); C76H125N23O19S (MW=1733.0\u2005g\u2009mol\u22121).Auxiliary Cleavage from Model Ligation Products: Lyophilized ligation products were dissolved in the auxiliary cleavage mixture to a final concentration of 1\u2005mM in plastic tubes. The closed lid was punctured to allow supply of oxygen. Note: Hydrazine is a known oxygen scavenger and auxiliary removal does not tolerate the presence of hydrazine.Removal of Auxiliary from Ligation Product AN (51AN): 500\u2005nmol of ligated peptide 42AN was incubated in the cleavage mixture for 3\u2005h. Yield: 250\u2005nmol, 50\u2009%. UPLC\u2010MS: tR=4.44\u2005min ; m/z=770.7 (C67H108N22O20 (M+2H)2+, calcd.: 770.4), 514.2 (C67H109N22O20 (M+3H)3+, calcd.: 513.9); C67H106N22O20 (MW=1538.8\u2005g\u2009mol\u22121).Removal of Auxiliary from Ligation Product AV (51AV): 500\u2005nmol of ligated peptide 42AV was incubated in the cleavage mixture for 6\u2005h. Yield: 180\u2005nmol, 36\u2009%. UPLC\u2010MS: tR=3.85\u2005min (then 3\u201330\u2009% B in 6\u2005min); m/z=763.2 (C68H111N21O19 (M+2H)2+, calcd.: 762.9), 509.2 (C68H112N21O19 (M+3H)3+, calcd.: 508.9); C68H109\u2005N21O19 (MW=1523.8\u2005g\u2009mol\u22121).Removal of Auxiliary from Ligation Product LN (51LN): 260\u2005nmol of ligated peptide 42LN was incubated in the cleavage mixture for 3\u2005h. Yield: 104\u2005nmol, 40\u2009%. UPLC\u2010MS: tR=5.16\u2005min ; m/z=791.7 (C70H114N22O20 (M+2H)2+, calcd.: 791.4), 528.1 (C70H115N22O20 (M+3H)3+, calcd.: 527.9); C70H112N22O20 (MW=1580.8\u2005g\u2009mol\u22121).Removal of Auxiliary from Ligation Product LV (51LV): 300\u2005nmol of ligated peptide 42AN was incubated in the cleavage mixture for 6\u2005h. Yield: 109\u2005nmol, 36\u2009%. UPLC\u2010MS: tR=4.14\u2005min (3\u201330\u2009% B in 6\u2005min); m/z=784.5 (C71H117N21O19 (M+2H)2+, calcd.: 783.9), 523.1 (C71H118N21O19 (M+3H)3+, calcd.: 523.0); C71H115N21O19 (MW=1566.8\u2005g\u2009mol\u22121).4 Thioester EIQALEEENAQLEQENAALEEEIAQLEY\u2010(APTTSTTS): The peptide was synthesized on a rink amide resin pre\u2010loaded with Hnb\u2010(StBu)Cys\u2010Gly via automated microwave assisted peptide\u2010synthesis . The peptide resin was treated with H2N\u2212OH\u2009\u22c5\u2009HCl , washed and deprotected with TFA\u2009:\u2009phenol\u2009:\u2009H2O\u2009:\u2009TIS (88\u2009:\u20095\u2009:\u20095\u2009:\u20092 v/v/v/v) for 2\u2005h. The precipitated crude peptide was shaken at 37\u2009\u00b0C in an aqueous buffer containing 10\u2005vol.% MPA. The reactions was quenched with 10\u2005vol.% TFA. Preparative HPLC: 15\u201335\u2009% B in 40\u2005min. Yield: 0.43\u2005\u03bcmol, 31\u2009%, UPLC: tR=5.95\u2005min (15\u201335\u2009% B in 8\u2005min). The product was submitted to ligation without further characterization.4\u2010KIAQLKQKIQALKQENQQLEEENAALEY MMPyE\u2010(APTTSTTS): Synthesis was performed on a rink amide resin via automated microwave assisted peptide\u2010synthesis . For reductive alkylation 10\u2005equiv 40 and 30\u2005equiv NaCNBH3 were allowed to react for 24\u2005h. Preparative HPLC: 15\u201335\u2009% B in 40\u2005min. Yield: 771\u2005nmol, 57\u2009%. UPLC: tR=4.27\u2005min (15\u201335\u2009% B in 8\u2005min). The product was submitted to the ligation without further characterization.Ligation of 53 and 54 (\u219255): Peptide thioester 53 (315\u2005nmol) and auxiliary peptide 54 (210\u2005nmol) were dissolved in ligation buffer and allowed to react for 120\u2005h. Yield: 53\u2005nmol, 25\u2009%. UPLC: tR=5.87\u2005min (15\u201335\u2009% B in 8\u2005min). The product was submitted to auxiliary cleavage without further characterization.8\u2010P4 (56)P1\u2010Muc5AC: Ligation product 55 (18\u2005nmol) was incubated in the cleavage mixture for 4\u2005h. Preparative HPLC: 15\u201335\u2009% B in 40\u2005min. Yield: 9.5\u2005nmol, 53\u2009%. UPLC: tR=6.6\u2005min (15\u201335\u2009% B in 8\u2005min); HRMS: m/z=1786.9876 (C522H862N139O214 (M+11H)7+, calcd.: 1786.7282), 1563.5361 (C522H863N139O214 (M+11H)8+, calcd.: 1563.5131), 1389.9659 (C522H864N139O214 (M+11H)9+, calcd.: 1389.9013), 1251.0593 (C522H865N139O214 (M+11H)10+, calcd.: 1251.0119), 1137.4254 (C522H866N139O214 (M+11H)11+, calcd.: 1137.3751), 1042.6994 (C522H867N139O214 (M+12H)12+, calcd.: 1042.6778).Synthesis of phosphorylated MMPyE\u2010CTD Peptide 57: Synthesis was performed on a Rink amide resin via MW\u2010SPPS . For reductive alkylation 7\u2005equiv 40 and 16\u2005equiv NaCNBH3 were allowed to react for 18\u2005h. The peptide was cleaved for 6\u2005h with TFA:H2O:phenol:EDT protecting group to avoid formation of cleavage side\u2010product). Preparative HPLC: 3\u201340\u2009% B in 40\u2005min. Yield: 3.46\u2005\u03bcmol, 35\u2009%. UPLC\u2010MS: tR=3.45\u2005min (3\u201330\u2009% B in 4\u2005min); m/z=1172.4, (C104H148N23O37S (M+3H)3+, calcd.: 1172.5), m/z=783.3, (C104H148N23O37S (M+4H)4+, calcd.: 783), C104H150N23O46P3S (MW=2343.01\u2005g\u2009mol\u22121). UPLC\u2010MS: tR=2.53\u2005min (3\u201330\u2009% B in 4\u2005min); m/z=862.0, (C104H150N23O46P3S (M+3H)3+, calcd.: 861.6), C104H150N23O46P3S (MW=2581.9\u2005g\u2009mol\u22121).6\u2010[(CH2)2O]6CH2CO\u2010(YpSPTSPS)3 Hydrazide (58)His: The peptide was assembled on a chlorotriyl resin pre\u2010loaded with Fmoc\u2010hydrazine via automated solid\u2010phase peptide synthesis . After introduction of the PEG6\u2010chain (Fmoc\u2010HN\u2010[(CH2)2O]6\u2010COOH (5\u2005equiv), HATU and DIPEA (15\u2005equiv) in DMF), capping and coupling of six histidines, the peptide was cleaved with TFA:H2O:phenol:EDT for 3\u2005h. Preparative HPLC 3\u201340\u2009% B in 40\u2005min. Yield: 1.86\u2005\u03bcmol, 19\u2009%; 58: 2.15\u2005\u03bcmol, 22\u2009%. UPLC\u2010MS (58): tR=2.08\u2005min (3\u201050\u2009% B in 4\u2005min); m/z=1197.3, (C147H213N42O58P3 (M+3H)3+, calcd.: 1196.8), 898.8 (C147H210N42O49 (M+4H)4+, calcd.: 897.9), 719.2 (C147H210N42O49 (M+5H)5+, calcd.: 718.5).Formation of Thioester 59, Ligation with 57\u2009and Removal of MMPyE Group (60): Peptide hydrazide 58 was dissolved in H2O containing 200\u2005mM MPAA and 6\u2005M Gdn HCl (pH=3) and acetylacetone was added. After 1\u2005h reaction at room temperature, the solution was transferred to a mixture of auxiliary peptide 57 in 100\u2005mM TCEP, 200\u2005mM Na2HPO4 and 6\u2005M Gdn HCl (pH=8.5). NaOH (5\u2005M) was added carefully by alternating addition over the vessel cap followed by vortexing to adjust the pH to 6.3. After 120\u2005h, the ligation was quenched by the addition of 5\u2005\u03bcl hydrazine solution (81\u2009% in H2O) and 10\u2005\u03bcl of TCEP (1\u2005M in H2O) and the resulting mixture was subjected to an Amicon Ultra Mass Filter from Merck Millipore and washed twice. After lyophilization, the crude peptide mixture was dissolved in 100\u2005\u03bcl phosphate buffer and purified via a His\u2010Tag kit using Ni2+ Agarose\u2010NTA beads. The resulting peptide solution was lyophilized and used i without further purification. The peptide mixture obtained after lyophilization was dissolved in 200\u2005\u03bcL of the auxiliary removal mixture for 24\u2005h. Preparative HPLC: 3\u201340\u2009% B in 40\u2005min. Yield: 18\u2005nmol, 4\u2009% over 3 steps. UPLC\u2010MS: tR=5.47\u2005min (5\u201030\u2009% B in 8\u2005min); m/z=1195.1 (C243H350N62O103P6 (M+5H)5+, calcd.: 1195.1), 996.9 (C243H350N62O103P6 (M+6H)6+, calcd.: 996.0), m/z=854.1 (C243H350N62O103P6 (M+7H)7+, calcd.: 853.9) m/z=745.9 (C243H350N62O103P6 (M+8H)8+, calcd.: 747.3) m/z=664.3 (C243H350N62O103P6 (M+9H)9+, calcd.: 664.4), (C243H350N62O103P6 (MW=5970.3).Synthesis of MMPyE\u2010peptide 62, Formation of Thioester 63\u2009and Cyclization (64): Microwave assisted peptide\u2010synthesis was performed on a pre\u2010loaded chlorotrityl\u2010tentagel resin . For reductive alkylation 7\u2005equiv of 40 and 16\u2005equiv of NaCNBH3 were allowed to react for 18\u2005h. Approx, 1.0\u2005\u03bcmol MMPyE\u2010peptide 62 was cleaved from the resin with hexafluoroisopropanol/DCM (7\u2009:\u20093 v/v) and thioester 63 was formed according to the general procedure . The peptide was cleaved for 18\u2005h with TFA, H2O, phenol, thioanisole, EDT (82\u2009:\u20095\u2009:\u20095\u2009:\u20095\u2009:\u20093 v/v/v/v/v) and incubated in ligation buffer at 37\u2009\u00b0C for 2\u2005h. Preparative HPLC: 20\u201350\u2009% B in 40\u2005min. Yield: 89\u2005nmol, 9\u2009%. UPLC\u2010MS: tR=4.88\u2005min (20\u201350\u2009% B in 8\u2005min); m/z=1138.2, (C109H150N24O28S (M+2H)2+, calcd.: 1138.0), 758.9 (C109H151N24O28S (M+3H)3+, calcd.: 759.0), C109H148N24O28S (MW=2274.55\u2005g\u22c5mol\u22121).Cyclopeptide 65: 55\u2005nmol of ligation product 64 was incubated in the cleavage mixture for 12\u2005h. Preparative HPLC: 20\u201350\u2009% B in 40\u2005min. Yield: 24\u2005nmol, 44\u2009%. UPLC\u2010MS: tR=3.93\u2005min (20\u201350\u2009% B in 8\u2005min); m/z=1054.42 (C101H141N23O27 (M+2H)2+, calcd.: 1054.02), 703.01 (C101H142N23O27 (M+3H)3+, calcd.: 703.35), C101H139N23O27 (MW=2107.32). MALDI\u2010MS: m/z=2104 (C101H140N23O27 (M+H)+, calcd.: 2108).More detailed procedures and characterization data is provided in the Supporting Information.The authors declare no conflict of interest.1As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "In the crystal, these components are organized in infinite zigzag chains via inter\u00admolecular hydrogen bonds. Weak inter\u00adactions between the chains lead to a three-dimensional network.The title salt consists of three components, comprising one di\u00adbenzyl\u00adammonium cation, [(C 14H16N+\u00b7C6H7AsNO3\u2212\u00b7H2O or [(C6H5CH2)2NH2][H2NC6H4As(OH)O2]\u00b7H2O, (I), was synthesized by mixing an aqueous solution of (4-amino\u00adphenyl)\u00adarsonic acid with an ethano\u00adlic solution of di\u00adbenzyl\u00adamine at room temperature. Compound I crystallizes in the monoclinic P21/c space group. The three components forming I are linked via N\u2014H\u22efO and O\u2014H\u22efO inter\u00admolecular hydrogen bonds, resulting in the propagation of an infinite zigzag chain. Additional weak inter\u00adactions between neighbouring chains, such as \u03c0\u2013\u03c0 and N\u2014H\u22efO contacts, involving phenyl rings, \u2013NH2 and \u2013As(OH)O3 functions, and H2O, respectively, lead to a three-dimensional network.The title salt, C This compound was employed for pharmaceutical applications, in particular against trypanosomal infection. Subsequently, in the early 20th century, Paul Ehrlich was inspired by this work to develop a new organoarsenic derivative, called Arsphenamine or Salvarsan has been determined 2NH2][H2NC6H4As(OH)O2]\u00b7H2O (I).From a structural point of view, the crystal structure of phenyl\u00adarsonic acid was first solved in the early 1960s 2NH2]+, one hydrogen (4-amino\u00adphen\u00adyl)arsonate anion [H2NC6H5As(OH)O2]\u2212 and one water mol\u00adecule of solvation. The three components of I are linked together through inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds. The As atom of the anion is bonded to three O atoms and one carbon atom of the phenyl ring, describing a slightly distorted tetra\u00adhedral geometry . The As\u2014O bonds exhibit two distinct lengths: As\u2014O1 = 1.7267\u2005(10)\u2005\u00c5, and As\u2014O2 = 1.6730\u2005(10)\u2005\u00c5 and As\u2014O3 =1.6699\u2005(10)\u2005\u00c5, which can be considered to be identical. The As\u2014O1 distance is consistent with the presence of a hydroxyl group \u2005\u00c5; symmetry code: (iv) \u2212x, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01, Table\u00a01et al., 2002para position to the arsonate function. Both functional groups are contained in the plane of the phenyl ring. The negative charge of [H2NC6H4As(OH)O2]\u2212 is compensated by the presence of one di\u00adbenzyl\u00adammonium cation, [(C6H5CH2)2NH2]+, whose NH2+ group is hydrogen bonded to the oxygen atom O3 of the arsonate function . Moreover, the di\u00adbenzyl\u00adammonium cation shows a syn\u2013anti conformation, displaying C\u2014C\u2014N\u2014C torsion angles of 57.65\u2005(16)\u00b0 and \u2212178.14\u2005(11)\u00b0, which are in the range of previous examples of X-ray structures involving [(C6H5CH2)2NH2]+ \u2005\u00c5; symmetry code: (v) 1\u00a0+\u00a0x, y, z] completes the composition of salt I. From a spectroscopic point of view, the infrared spectrum of I (ATR mode) highlights \u03bd(As\u2014C) and \u03bd(As\u2014O) absorption bands, which are characteristic of the arsonate function .The asymmetric unit of the title salt, which is depicted in Fig.\u00a013.I:At the supra\u00admolecular stage, two levels of organization can be observed in the crystal structure of a-axis direction resulting from the hydrogen-bonding inter\u00adactions \u00b0], with the oxygen atoms O3 of two arsonate moieties and the O1 oxygen atom of an adjacent \u2013As(OH)O2 function [N2\u2014H2B\u22efO1ii = 3.0769\u2005(17)\u2005\u00c5] (symmetry codes as in Table\u00a01(ii) The association of chains leading to a three-dimensional network and resulting from a combination of weak inter\u00adactions Fig.\u00a04. Two typ4.et al., 20166H5As(OH)O2\u2212. To our knowledge, eleven examples including this fragment have already been identified: ammonium 4-nitro\u00adphenyl\u00adarsonate 2NH2]+, 117 hits incorporating such an entity were found in the Cambridge Structural Database.A search of the Cambridge Structural Database 2O] was prepared according to a previous work and arsenic acid (As(OH)3O). The title salt was obtained by neutralization of an aqueous solution (20\u2005mL) of (4-amino\u00adphen\u00adyl)arsonic acid with di\u00adbenzyl\u00adamine ((C6H5CH2)2NH) dissolved in 20\u2005mL of ethanol. The mixture was stirred for about two\u2005h at room temperature (301\u2005K). After three days of slow solvent evaporation, colourless prism-shaped crystals of [(C6H5CH2)2NH2][H2NC6H4As(OH)O2]\u00b7H2O , suitable for an X-ray crystallographic analysis, were collected from the solvent (m.p. 393\u2005K). FT\u2013IR : 3447, 3304, 3187, 1595, 1501, 1454, 1096, 923, 878, 825,752, 735, 695. Elemental analysis \u2013 analysis calculated for C20H23N2O3As\u00b70.25H2O (418.83), salt I partially dehydrated: C, 57.35; H, 5.66; N, 6.69; O, 12.41; found: C, 57.82; H, 5.61; N, 6.62; O, 12.37%.All chemicals were purchased from Sigma-Aldrich (Germany) and used without any further purification. (4-Amino\u00adphen\u00adyl)arsonic acid [H6.Uiso(H) = 1.5Ueq(O). C-bound hydrogen atoms were placed at calculated positions [C\u2014H = 0.95\u2005\u00c5 (aromatic) or 0.99\u2005\u00c5 (methyl\u00adene group)] and H atoms of the NH2 and OH terminal groups were placed geometrically and refined as riding with Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698902300837X/dj2065sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698902300837X/dj2065Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902300837X/dj2065Isup3.cmlSupporting information file. DOI: 2297206CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The magnesium cation exhibits a distorted octa\u00adhedral coordination with two bidentate di\u00admeth\u00adoxy\u00adethane mol\u00adecules and a dinuclear aluminate anion, coordinated to Mg2+via oxido and hydroxido units. The anion is an oxido-bridged species, [HOAl(hfip)2(\u03bc-O)Al(hfip)3]2\u2013, with one Al3+ cation tetra\u00adhedrally coordinated by an oxido (O2\u2013) anion, a hydroxido anion, and two hfip groups, whereas the second Al3+ cation is coordinated by the oxido anion and three hfip groups.Partial hydrolysis of a sample of [Mg(dme) The nearly right angle involves the oxido and hydroxido groups (O\u2014Al1\u2014OH). The anion coordinates to the magnesium cation via hydroxido and oxido units, thus making these ligands \u03bc- and \u03bc3-bridges, respectively, resulting in an Mg1\u22efAl1 distance of 2.8074\u2005(3)\u2005\u00c5. The angles at the hydroxido and oxido bridges between Al1 and Mg1 are similar , whereas the angles involving \u03bc3-oxido and Al2 are more obtuse . The Mg2+ cation has a distorted octa\u00adhedral coordination with cis-O\u2014Mg\u2014O angles ranging from 76.40\u00b0 (2) to 101.00\u2005(2)\u00b0 and trans-O\u2014Mg\u2014O angles in the range 164.77\u2005(3)\u2013168.38\u2005(3)\u00b0. The O\u2014Mg\u2014O bite angles of the dme ligands [76.40\u2005(3) and 77.08\u2005(3)\u00b0] and the anion [76.99\u2005(2)\u00b0] are nearly identical.The title compound crystallizes in the monoclinic space group ly Fig.\u00a01. In the \u00c5] Fig.\u00a01. The tet2(CF3SO3)2] 2 \u20131.7444\u2005(10)\u2005\u00c5 in NMe4[Al(hfip)4] 4] , 2.1146\u2005(12)\u2005\u00c5, 77.49\u2005(5)\u00b0 in sample 3][Al(hfip)4]2 + 2H2O \u2192 [Mg(dme)2{HOAl(hfip)2OAl(hfip)3}] + dme + 3hfipH.[Mg(dme)et al., 2010The crystal data, data collection, and structure refinement details are summarized in Table\u00a0210.1107/S2414314623007162/hb4443sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314623007162/hb4443Isup2.hklStructure factors: contains datablock(s) I. DOI: 2156683CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two compounds have a structural overlap r.m.s. deviation of 0.27\u2005\u00c5. The pyran and seven-membered cyclo\u00adheptene rings in both structures adopt boat and boat-sofa conformations, respectively. Intra- and inter\u00admolecular C\u2014H\u22efO hydrogen bonds are responsible for the consolidation of the crystal packing of both mol\u00adecules. In addition to this, weak C\u2014H\u22ef\u03c0 inter\u00adactions are also observed. The inter\u00admolecular inter\u00adactions were qu\u00adanti\u00adfied and analysed using Hirshfeld surface analysis.The title compounds, C It is the fundamental component of many beneficial alkaloids, including atropine, scopolamine, and cocaine, whose derivatives are important in the treatment of neurological and psychiatric conditions such depression and panic disorder q2 = 1.021\u2005(2) and q3 = 0.391\u2005(2)\u2005\u00c5 in (I)q2 = 1.053\u2005(2) and q3 = 0.374\u2005(2)\u2005\u00c5 in (II)q2 = QT = 0.185\u2005(2)\u2005\u00c5 and \u03c6 = 43.9\u2005(1)\u00b0 [in (I)] and q2 = QT = 0.087\u2005(1)\u2005\u00c5 and \u03c6 = 47.1\u2005(1)\u00b0 [in (II)]. The cyclo\u00adhexene ring (C15\u2013C20) has a distorted sofa conformation in both (I)sC(C17) asymmetry parameters in both compounds has a boat-sofa conformation, with puckering parameters , see Fig.\u00a04In the crystal of (I)via C\u2014H\u22efO inter\u00admolecular inter\u00adaction , see Fig.\u00a04In the crystal of (II)if Fig.\u00a05. The dim4.Crystal Explorer 21 et al., 2021To further characterize the inter\u00admolecular inter\u00adactions in the title compound, we carried out a Hirshfeld surface (HS) analysis Compound (II)6.Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698902300275X/zn2026sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S205698902300275X/zn2026Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S205698902300275X/zn2026IIsup3.hklStructure factors: contains datablock(s) II. DOI: 2251053, 2251052CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The layers are further connected by \u03c0\u2013\u03c0 stacking inter\u00adactions.In the crystal of the title com\u00adpound, C\u2014H\u22efO hydro\u00adgen bonds link the mol\u00adecules, enclosing 14H11NO3, the di\u00adhydro\u00adquinoline core deviates slightly from planarity, indicated by the dihedral angle of 1.07\u2005(3)\u00b0 between the two six-membered rings. In the crystal, layers of mol\u00adecules almost parallel to the bc plane are formed by C\u2014H\u22efO hydro\u00adgen bonds. These are joined by \u03c0\u2013\u03c0 stacking inter\u00adactions. A Hirshfeld surface analysis revealed that the most important contributions to the crystal packing are from H\u22efH (36.0%), H\u22efC/C\u22efH (28.9%) and H\u22efO/O\u22efH (23.5%) inter\u00adactions. The evaluation of the electrostatic, dispersion and total energy frameworks indicates that the stabilization is dominated by the dispersion energy contribution. Moreover, the mol\u00adecular structure optimized by density functional theory (DFT) at the B3LYP/6-311G level is com\u00adpared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.In the title mol\u00adecule, C Moreover, a Hirshfeld surface analysis and inter\u00adaction energy and energy framework calculations were performed. The mol\u00adecular structure optimized by density functional theory (DFT) at the B3LYP/6-311G level is com\u00adpared with the experimentally determined mol\u00adecular structure in the solid state.2.A (C1\u2013C5/N1) and B (C4\u2013C9) rings. Atoms O1, O2, O3, C10, C13 and C14 are \u22120.1294\u2005(11), 0.1907\u2005(12), \u22120.2708\u2005(15), 0.0177\u2005(14), \u22120.0267\u2005(13) and 0.0953\u2005(23)\u2005\u00c5 from the least-squares plane of the A ring. The O2\u2014C13 [1.3123\u2005(17)\u2005\u00c5] and O3\u2014C13 [1.1955\u2005(16)\u2005\u00c5] distances in the ester group indicate localized single and double bonds, rather than delocalized bonding arrangements. The O2\u2014C13\u2014O3 bond angle [122.55\u2005(12)\u00b0] seems to be slightly increased with respect to that present in a free acid bis\u00ad(nicotinamide-\u03baN1)zinc(II) rings to form a triperiodic network.In the crystal, C\u2014H\u22efO hydro\u00adgen bonds Table\u00a01 link thene Fig.\u00a02. These l4.CrystalExplorer ] and those delineated into H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH, C\u22efC, C\u22efO/O\u22efC, H\u22efN/N\u22efH, C\u22efN/N\u22efC and N\u22efO/O\u22efN contacts \u2013(i), respectively, together with their relative contributions to the Hirshfeld surface. The most important inter\u00adaction is H\u22efH, contributing 36.0% to the overall crystal packing, which is reflected in Fig.\u00a06b) as widely scattered points of high density due to the large hydro\u00adgen content of the mol\u00adecule with the tip at de = di = 1.22\u2005\u00c5. In the absence of C\u2014H\u22ef\u03c0 inter\u00adactions, the pair of characteristic wings resulting in the fingerprint plot delineated into H\u22efC/C\u22efH contacts [Fig.\u00a06c)] have a 28.9% contribution to the HS, with the tips at de + di = 2.68\u2005\u00c5. The pair of the scattered points of spikes resulting in the fingerprint plot delineated into H\u22efO/O\u22efH contacts [Fig.\u00a06d)], with a 23.5% contribution to the HS, has an almost symmetric distribution of points, with the tips at de + di = 2.44\u2005\u00c5. The C\u22efC contacts [Fig.\u00a06e)] appear as an arrow-shaped distribution of points and have a contribution of 7.0% to the HS with the tip at de = di = 1.69\u2005\u00c5. The tiny spikes of C\u22efO/O\u22efC contacts [Fig.\u00a06f)], with a 2.5% contribution to the HS, are visible at de + di = 3.58\u2005\u00c5. Finally, the H\u22efN/N\u22efH [Fig.\u00a06g)], C\u22efN/N\u22efC [Fig.\u00a06h)] and N\u22efO/O\u22efN [Fig.\u00a06i)] contacts contribute 1.4, 0.4 and 0.4%, respectively, to the HS.In order to visualize the inter\u00admolecular inter\u00adactions in the crystal of the title com\u00adpound, a Hirshfeld surface (HS) analysis \u2013(c), respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions play the major role in the crystal packing is the sum of electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange\u2013repulsion (Erep) energies , Edis (green cylinders) and Etot (blue cylinders), and are shown in Figs.\u00a08a)\u2013(c). The evaluation of the electrostatic, dispersion and total energy frameworks indicates that in the title com\u00adpound the stabilization is dominated by the dispersion energy contribution.Using 6.GAUSSIAN09 using the standard B3LYP functional and the 6311G basis set derived from the conceptual DFT calculations are given in Table\u00a03E = ELUMO \u2212 EHOMO] , with layer-by-layer connections approximately parallel to and tetra-n-butyl\u00adammonium bromide . The reaction mixture was stirred at room tem\u00adper\u00ada\u00adture in DMF for 6\u2005h. After removal of the formed salts, the solvent was evaporated under reduced pressure and the residue obtained was dissolved in di\u00adchloro\u00admethane. The organic phase was dried over Na2SO4 and then concentrated in vacuo. A pure com\u00adpound was obtained after recrystallization from di\u00adchloro\u00admethane/hexane (2:3 v/v).To a solution of methyl 2-oxo-1,2-di\u00adhydro\u00adquinoline-4-car\u00adbox\u00adyl\u00adate (4.47\u2005mmol) in 10\u2005ml of di\u00admethyl\u00adformamide (DMF) were added propargyl bromide (9.83\u2005mmol), K10.1107/S2056989023007557/wm5688sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989023007557/wm5688Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023007557/wm5688Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023007557/wm5688Isup4.cdxSupporting information file. DOI: 2291603CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "G\u03b3- and A\u03b3-globin genes were examined by DNA analysis and rapid diagnosis of HPFH mutations were developed by PCR-based methods. Twenty subjects with five different mutations were identified including three known mutations, \u2212\u00a0202 A\u03b3 (C>T) (n\u2009=\u20093), \u2212\u00a0196 A\u03b3 (C>T) (n\u2009=\u20093), and \u2212\u00a0158 A\u03b3 (C>T) (n\u2009=\u200912), and two novel mutations, \u2212\u00a0117 A\u03b3 (G>C) (n\u2009=\u20091) and \u2212\u00a0530 G\u03b3 (A>G) (n\u2009=\u20091). Interaction of the \u2212\u00a0117 A\u03b3 (G>C) and Hb E (HBB:c.79G>A) resulted in elevation of Hb F to the level of 13.5%. Two plain heterozygous subjects with \u2212\u00a0530 G\u03b3 (A>G) had marginally elevated Hb F with 1.9% and 3.0%, whereas the proband with homozygous \u2212\u00a0530 G\u03b3 (A>G) had elevated Hb F of 11.5%. Functional prediction indicated that the \u2212\u00a0117 A\u03b3 (G>C) and \u2212\u00a0530 G\u03b3 (A>G) mutations dramatically alter the binding of transcription factors to respective \u03b3-globin gene promotors, especially the CCAAT and GATA-1 transcription factors. Diverse heterogeneity of non-deletional HFPH with both known and new mutations, and complex interactions of them with other forms of thalassemia are encountered in Thai population.High Hb F determinants are genetic defects associated with increased expression of hemoglobin F in adult life, classified as deletional and non-deletional forms. We report the first description of non-deletional hereditary persistence of fetal hemoglobin (HFPH) in Thailand. Study was done on 388 subjects suspected of non-deletional HPFH with elevated Hb F expression. Mutations in the In Thailand, high prevalence of hemoglobinopathies has been documented including 20\u201330% of \u03b1-thalassemia, 3\u20139% of \u03b2-thalassemia, 20\u201330% of Hb E (HBB:c.79G>A), 1\u20138% of Hb Constant Spring (HBA2:c.427T>C) and Hb Pakse\u2032 (HBA2:c.429A>T), and other structural Hb variants and high Hb F determinants. These lead to diverse heterogeneity and complex thalassemia syndromes in the country5. Accurate diagnosis of these hemoglobinopathies is important for promoting appropriate management, genetic counseling, and a prevention and control program of thalassemia in the region.The inherited disorders of hemoglobin (Hb) or hemoglobinopathies are the commonest human monogenic diseases, found in about 7% of the world population. These can be broadly classified into 3 groups, structural Hb variants, thalassemia and hereditary persistence of fetal hemoglobin (HPFH)7. Since co-inheritance of these high Hb F determinants in \u03b2-thalassemia disease has an ameliorating effect on the disease severity, identification of them in routine practice is important. At the molecular level, they could be classified into deletional and non-deletional high Hb F determinants. While the former involves several large DNA deletions removing \u03b4- and \u03b2-globin genes, the latter is usually caused by point mutations on G\u03b3- or A\u03b3-globin gene promoters affecting the binding of related transcription factors5.During early childhood, the level of Hb F normally declines to less than 1\u20132% of total Hb. The level of Hb F is therefore less than 1\u20132% in normal adult. High Hb F determinants are a group of genetic defects associated with increased expression of Hb F in adult life. The conditions include \u03b4\u03b2-thalassemia, \u03b3\u03b4\u03b2-thalassemia and hereditary persistence of fetal hemoglobin (HPFH). While \u03b4\u03b2-thalassemia and \u03b3\u03b4\u03b2-thalassemia are usually associated with elevated Hb F and hypochromic microcytic red blood cell indices, HPFH is associated with higher Hb F expression with normal red blood cells. Homozygous or compound heterozygous for \u03b4\u03b2-thalassemia and \u03b2-thalassemia may have clinical phenotype of non-transfusion-dependent thalassemia or transfusion-dependent thalassemia. In contrast, homozygous HPFH or compound heterozygous HPFH and \u03b2-thalassemia usually have no clinical symptom. However, these clinical phenotypes are generally overlapped which renders their differentiation in routine setting difficult unless DNA analysis is performed0-thalassemia (12.6\u00a0kb deletion) (NG_000007.3:g.64383_76994), Indian deletion-inversion G\u03b3(A\u03b3\u03b4\u03b2)0-thalassemia , Chinese A\u03b3\u03b4\u03b20-thalassemia (78.9\u00a0kb deletion) (NG_000007.3:g.48795_127698del), \u03b4\u03b20-thalassemia (11.3\u00a0kb deletion) (NG_000007.3:g.60045_71313delinsTACATTAAGAGATACCTTAATG), Siriraj A\u03b3\u03b4\u03b20-thalassemia (~\u2009118\u00a0kb deletion) (AC104389.8:g.52507_165744), Thai deletion-inversion-insertion A\u03b3\u03b4\u03b20-thalassemia and Vietnamese/SEA HPFH (27\u00a0kb deletion) (NG_000007.3:g.64384_76993del)10. However, non-deletional form of HFPH has rarely been described. This study provides for the first time the molecular description of non-deletional HFPH in Thailand including known and novel mutations affecting the CCAAT and GATA-1 transcription factors binding.In Thailand, several deletional forms of \u03b4\u03b2-thalassemia and HPFH have been documented. These included HPFH-6 (79.3\u00a0kb deletion) (NG_000007.3:g.45595_124872), \u03b4\u03b210. These included subjects with normal \u03b2-globin gene but Hb F\u2009\u2265\u20091.0% (n\u2009=\u200981), Hb E heterozygotes with Hb F\u2009\u2265\u20091.0% (n\u2009=\u2009287), \u03b2-thalassemia carriers with Hb F>10% (n\u2009=\u20096), and homozygous Hb E with Hb F>20% (n\u2009=\u200914). For comparison, normal individuals with Hb F\u2009<\u20091.0% (n\u2009=\u200960) were also analyzed. Hematological data is routinely recorded on standard automated blood cell counter. Hb analysis is performed using automated capillary electrophoresis system or automated high-performance liquid chromatography (HPLC) .This study was conducted in accordance with the Declaration of Helsinki and ethical approval of this study protocol was obtained from the Institutional Review Board of Khon Kaen University, Thailand (HE652154). Left-over blood specimens of 388 subjects suspected of non-deletional HPFH were selectively recruited at our routine thalassemia diagnostic service at Khon Kaen University, Thailand. They were all negative for deletional forms of high Hb F determinants previously found in ThailandSEA, --THAI, -\u03b13.7, -\u03b14.2, Hb Constant Spring, and Hb Pakse\u2032), \u03b2-thalassemia, deletional high Hb F determinants, and Kr\u00fcppel-like factor 1 (KLF1) mutations found in Thai population were identified by PCR-based methods as described previously14. G\u03b3- and A\u03b3-globin promoter mutations were examined by DNA sequencing on ABI PRISM\u2122 3730 XL analyzer using primers F35 and \u03b3515, and F229 and \u03b33516. Sequences of all primers used in this study are listed in the Supplementary Table Common \u03b1-thalassemia (--G\u03b3- globin promoter (C>T) (HBG2:c.\u2212\u00a0211C>T) and \u2212\u00a0158 A\u03b3-globin promoter (C>T) (HBG1:c.\u2212\u00a0211C>T) was developed as shown in Supplementary Fig. G\u03b3- and A\u03b3-globin gene promoters was done using primers \u03b3415 and \u03b35 (577\u00a0bp in length), and F22 & \u03b35 (639\u00a0bp in length), respectively. With this assay, the amplified product was digested to completion with XmnI restriction enzyme (5\u2032-GAANN\u25bcNNTTC-3\u2032) . After digestion, the 577\u00a0bp of \u2212\u00a0158 G\u03b3-globin promoter with T allele [XmnI (+)] was digested into two fragments with 402\u00a0bp and 175\u00a0bp in lengths, and the 639\u00a0bp fragment for the \u2212\u00a0158 A\u03b3- globin promoter with T allele [XmnI (+)] was digested into two fragments with 464\u00a0bp and 175\u00a0bp in lengths. The \u2212\u00a0158 G\u03b3- and A\u03b3-globin promoters with C allele [XmnI (\u2212)] remain undigested16.The PCR-restriction fragment length polymorphism (PCR\u2013RFLP) assay for detection of \u2212\u00a0158 A\u03b3-globin (C>T) (HBG1:c.\u2212\u00a0255C>T) (HBG1:c.\u2212\u00a0249C>T) (HBG1:c.\u2212\u00a0170G>C) (HBG2:c \u2212\u00a0583A>G) , \u2212\u00a0196 A\u03b3-globin (C>T), and \u2212\u00a0117 A\u03b3-globin (G>C) mutations. Similarly, primers (F40 & \u03b35) were used to produce the 795\u00a0bp internal control fragment for \u2212\u00a0530 G\u03b3-globin (A>G) mutation. Specific primer pairs (F38 & F22), (F39 & F22), (G211 & \u03b35), and (F41 & F40) were used to generate specific fragments of 444, 450, 150, and 276 bps in length for the \u2212\u00a0202 A\u03b3-globin (C>T), \u2212\u00a0196 A\u03b3-globin (C>T), \u2212\u00a0117 A\u03b3-globin (G>C), and \u2212\u00a0530 G\u03b3-globin (A>G), respectively. The PCR mixture (50\u00a0\u03bcl) contains 50\u2013200\u00a0ng genomic DNA, 30\u00a0pmol each primer, 200\u00a0\u03bcM dNTPs and 1 unit Taq DNA polymerase in 10 mM\u00a0Tris-HCl (pH 8.3), 50 mM\u00a0KCl, 0.01% gelatin and 3 mM\u00a0MgCl2. The amplification reaction was carried out on a T-Personal Thermocycler . A total of 30 cycles after initial heating at 94\u00a0\u00b0C for 3\u00a0min was performed under the following PCR condition: 93\u00a0\u00b0C 30\u00a0s, 62\u00a0\u00b0C 30\u00a0s, and 72\u00a0\u00b0C for 1\u00a0min. The amplified PCR product was separated on 2.0% agarose gel electrophoresis and visualized under UV light after ethidium bromide staining.Allele-specific PCR assays were developed for rapid identification of four non-deletional HPFH mutations including \u2212\u00a0202 HincII, G\u03b3-HindIII, A\u03b3-HindIII, \u03c8\u03b2-HincII, 3\u2032\u03c8\u03b2-HincII, \u03b2-AvaII and 3\u2032\u03b2-BamHI were determined using PCR\u2013RFLP assays as described previously17.Seven polymorphic restriction sites on \u03b2-globin gene cluster including \u03b5-https://tfbind.hgc.jp/)18 was used to compare the binding affinity of transcription factors to the promoters of \u03b3-globin genes with wild-type sequences and their mutant counterparts including \u2212\u00a0117 A\u03b3 (G>A), \u2212\u00a0117 A\u03b3 (G>C), \u2212\u00a0114 A\u03b3 (C>T), \u2212\u00a0530 G\u03b3 (A>G), and \u2212\u00a0533 to \u2212\u00a0529 (-ATAAG) mutations.The online TFBIND software (G\u03b3- and A\u03b3-globin genes was selectively carried out in 177 subjects with Hb F>5.0%. Fourteen subjects (7.9%) with five mutations in the \u03b3-globin gene promoters were identified including three known mutations, \u2212\u00a0202 A\u03b3 (C>T) (n\u2009=\u20093), \u2212\u00a0196 A\u03b3 (C>T) (n\u2009=\u20093), and \u2212\u00a0158 A\u03b3 (C>T) (n\u2009=\u20096), and two novel mutations, \u2212\u00a0117 A\u03b3 (G>C) (n\u2009=\u20091) and \u2212\u00a0530 G\u03b3 (A>G) (n\u2009=\u20091). All mutations but the \u2212\u00a0530 G\u03b3 (A>G) were identified in heterozygotic form. Rapid PCR diagnosis based on PCR\u2013RFLP assays mutation on PCR\u2013RFLP based assay. No mutation was detected in 60 normal subjects with Hb F\u2009<\u20091.0%. Therefore, a total of 20 of 388 (5.1%) recruited subjects were found to carry HPFH mutations. These included in total \u2212\u00a0202 A\u03b3 (C>T) (n\u2009=\u20093), \u2212\u00a0196 A\u03b3 (C>T) (n\u2009=\u20093), \u2212\u00a0158 A\u03b3 (C>T) (n\u2009=\u200912), \u2212\u00a0117 A\u03b3 (G>C) (n\u2009=\u20091) and \u2212\u00a0530 G\u03b3 (A>G) (n\u2009=\u20091). As shown in Table A\u03b3 (C>T) HPFH mutation and Hb F 27.1% was a boy (2-yr-old) who was found to be a carrier of \u03b20-thalassemia (\u03b217(AAG>TAG), HBB:c.52A>T) and \u03b1+-thalassemia (3.7\u00a0kb deletion). This boy was therefore a triple heterozygote for A\u03b3-HPFH, \u03b20-thalassemia and \u03b1+-thalassemia, a hitherto undescribed complex condition. Extended family analysis indicated that the \u2212\u00a0158 A\u03b3 (C>T) mutation was detected in trans to the \u03b20-thalassemia gene. He had Hb A2 of 4.3%, still within diagnostic range for a \u03b2-thalassemia trait.Among 388 subjects recruited DNA sequencing of +-Thalassemia was detected in 8 of 20 HPFH subjects with relatively decreased Hb F levels as compared to those with the same HPFH mutations but without \u03b1-thalassemia. The overall hematological parameters, Hb analysis results as well as \u03b1- and \u03b2-globin genotypes of these 20 non-deletional HPFH were summarized in Table No mutation in both \u03b3-globin genes was identified in subjects with homozygous Hb E with unusually high Hb F level. Further screening for the KLF1 mutations previously described in Thai population showed that 3 of these 20 subjects with HPFH carried the KLF1 mutations i.e., H299D (NG_013087.1:g.6869C>G) (n\u2009=\u20091), T334R (NG_013087.1:g.7231C>G) (n\u2009=\u20091) and G176Afs*179 (NG_013087.1:g.6493_6499dupCGGCGCC) (n\u2009=\u20091). \u03b1G\u03b3 (A>G) novel mutation is located at the GATA-1 transcription factor binding motif of G\u03b3-globin gene promoter and is identified in homozygotic form, further family analysis was carried out as shown in Fig.\u00a0G\u03b3 (A>G) mutation. Apparently, heterozygosity for the \u2212\u00a0530 G\u03b3 (A>G) mutation is associated with normal hematological features and marginally elevated Hb F (1.9% in the mother and 3.0% in the sister). The father had alternatively reduced Hb F (0.3%) due to a co-inheritance of \u03b1+-thalassemia (3.7\u00a0kb deletion) since he was a double heterozygote for G\u03b3-HPFH and \u03b1+-thalassemia. In all members, the levels of Hb A2 were within normal range (2.4\u20132.6%). The proband\u2019s wife (27-year-old) was a patient with Hb E-\u03b2+-thalassemia with mild hypochromic microcytic anemia with Hb 10.1\u00a0g/dL, MCV 52.7 fL and MCH 18.6\u00a0pg. Therefore, they were at risk of having a child with compound G\u03b3-HPFH/\u03b2+-thalassemia or G\u03b3-HPFH/Hb E syndrome. In addition, screening for KLF1 mutations previously described in Thai population14 yielded negative results in all members. \u03b2-Globin gene haplotype analysis on 7 polymorphic restriction sites within \u03b2-globin gene cluster as described in the \u201cG\u03b3 (A>G) mutation was linked to the \u03b2-globin gene haplotype (-\u2009++-+-+) in this Thai family and pointed to the same origin of this G\u03b3-HPFH in the father and the mother. Appropriate genetic counselling was provided to the family members.Since the \u2212\u00a0530 0-thalassemia and A\u03b3\u03b4\u03b20-thalassemia. At least 7 different deletional forms of HPFH and \u03b4\u03b20-thalassemia have been characterized in Thailand10. In contrast, the molecular basis of non-deletional HPFH in Thailand has not been investigated. This study represents the first extensive study of non-deletional HPFH among Thai population. At present, around 30 non-deletional HFPH mutations on both G\u03b3- and A\u03b3-globin genes have been characterized worldwide. These include 15 mutations each on G\u03b3-globin and A\u03b3-globin genes 19. We have now reported for the first time in Thai population, five different HPFH mutations found in 20 subjects including four A\u03b3-HPFH and a G\u03b3-HPFH. Of these five HPFH mutations, three known mutations were \u2212\u00a0202 A\u03b3 (C>T) (n\u2009=\u20093), \u2212\u00a0196 A\u03b3 (C>T) (n\u2009=\u20093) and \u2212\u00a0158 A\u03b3 (C>T) (n\u2009=\u200912). Two novel mutations namely \u2212\u00a0117 A\u03b3 (G>C) and \u2212\u00a0530 G\u03b3 (A>G) were unexpectedly detected. As shown in Table In Thailand, we have extensively investigated the molecular basis of deletional form of HPFH and other high Hb F determinants including \u03b4\u03b2A\u03b3 (C>T) and the Italian/Chinese \u2212\u00a0196 A\u03b3 (C>T), are located within the G-rich area upstream of the \u2212\u00a0195 to \u2212\u00a0202 of \u03b3-globin gene. This DNA region is known to be the binding site of a ubiquitous transcription factor Sp1. As compared to the wild-type sequence, these two mutations decreased the binding of Sp1 on gel-shift experiment22. It has also been suggested that mutations in this G-rich sequence disrupt an intramolecular triplex proposed to be the binding site of a repressor, thereby increasing expression of \u03b3-globin gene23. Carriers of the \u2212\u00a0202 A\u03b3 (C>T) and \u2212\u00a0196 A\u03b3 (C>T) reported previously had Hb F in the ranges of 1.6\u20133.9% and 12\u201321%, respectively23. In our study, the three carriers of the \u2212\u00a0196 A\u03b3 (C>T) had as expected the average Hb F of 14.3\u2009\u00b1\u20091.9% (Table A\u03b3 (C>T) demonstrated higher Hb F levels with the average of 9.9\u2009\u00b1\u20091.6% as compared to the previously reported cases, possibly due to the presence of Hb E and the \u2212\u00a0158 G\u03b3 XmnI (\u2009+) in all Thai subjects. In addition, two of them also carried the KLF1 mutations (H299D & T334R) (case no. 2 & 3), known to be associated with increased Hb F expression in Hb E syndrome14.The Black \u2212\u00a0202 A\u03b3 (C>T) or the Cretan HPFH was originally described in three unrelated Greek adults with slightly increased Hb F level (2.9\u20135.1%) and normal hematological parameters. The \u2212\u00a0158 A\u03b3 (C>T) mutation has presumably resulted from a gene conversion event24. It is noteworthy that this form of HPFH was the most common one in Thai population, being detected in 12 of 20 Thai non-deletional HPFH subjects in this study (Table A\u03b3 (C>T) mutation on Hb F production can be raised. We found that this mutation is associated in cis with the \u2212\u00a0158 G\u03b3 XmnI (\u2009+) polymorphism in Thai population. It has been noted that a 240-kDa activator protein, a member of the CAAT/enhancer-binding proteins family, binds to \u2212\u00a0158 to -\u2212\u00a0161 nucleotides of \u03b3-globin gene promoter and induces expression of \u03b3-globin gene. This may explain the effect of \u2212\u00a0158 G\u03b3 XmnI (+) (C>T) and \u2212\u00a0158 A\u03b3 (C>T) to elevated \u03b3-globin gene expression25. While the \u2212\u00a0158 G\u03b3 XmnI (+) (C>T) polymorphism is associated with marginally elevated Hb F during erythropoietic stress, the \u2212\u00a0158 A\u03b3 (C>T) mutation is associated with higher Hb F expression with average of 7.8\u2009\u00b1\u20096.8% (Table A\u03b3-globin gene is more effective in competition with G\u03b3-globin gene for the Locus Control Region (LCR) in the adult stage25. The combined action of \u2212\u00a0158 A\u03b3 (C>T) in cis with \u2212\u00a0158 G\u03b3 (C>T) may lead further to higher Hb F production than having \u2212\u00a0158 G\u03b3 (C>T) alone24. In addition, study in Chinese and Thai subjects has identified that the \u2212\u00a0158 G\u03b3 XmnI (+) (C>T) was linked to the\u2009+\u200925 (G>A) polymorphism of A\u03b3-globin promoter (rs368698783), the binding motif of Ly-1 antibody reactive (LYAR) transcription factor. This polymorphism decreased the binding efficiency of the repressor of \u03b3-globin genes leading to increased Hb F production26. We found that all Thai subjects with \u2212\u00a0158 A\u03b3 (C>T) HPFH mutation also carried the \u2212\u00a0158 XmnI G\u03b3 (+) and the\u2009+\u200925 A\u03b3 (G>A) polymorphism. This should explain the higher Hb F production in these Thai subjects found with the HPFH mutation alone or in combinations several common hemoglobinopathies including Hb E, \u03b2-thalassemia, \u03b1-thalassemia, and KLF1 mutation (G176Afs*179). The lowest Hb F of 2.1% was found in a heterozygous subject with the \u2212\u00a0158 A\u03b3 (C>T) in combination with Hb E and \u03b1+-thalassemia (3.7\u00a0kb deletion). This is not unexpected since it has been known that co-inheritance of \u03b1-thalassemia can lead to the reduced Hb F level in several hemoglobinopathies29. Of interest is the finding of subject with complex interaction of the \u2212\u00a0158 A\u03b3 (C>T), \u03b1+-thalassemia (3.7\u00a0kb deletion), and \u03b20-thalassemia (\u03b217(AAG>TAG)) (Table 2 (4.3%) and as high as 27.1% Hb F 30. This is in contrast with the deletional form of high Hb F determinants in which co-inheritance with \u03b2-thalassemia is associated with normal Hb A2 level5. Although diagnosis of a \u03b2-thalassemia heterozygote with this complex interaction seen in the case no. 17 is not altered due to elevated Hb A2, an unusually increased Hb F (27.1%) in heterozygous \u03b2-thalassemia, as seen in Thai subject, might be a good marker for a co-inheritance of HPFH in \u03b2-thalassemia, requiring further investigation. We recommend therefore to investigate all cases of heterozygous \u03b2-thalassemia with Hb F higher than 2% for further investigation of a possible co-inheritance of HPFH. This is a very important and useful information at genetic counselling since co-inheritance of HPFH can ameliorate the severity of \u03b2-thalassemia disease3. The last case in this group of HPFH with \u2212\u00a0158 A\u03b3 (C>T) mutation is an adult male encountered with a homozygous for this mutation who had 7.6% Hb F. He also carried the heterozygosity for KLF1 mutation (p.G176Afs*179), and \u03b1+-thalassemia (3.7\u00a0kb deletion). Homozygosity for a \u2212\u00a0158 XmnI G\u03b3 (+) was not unexpectedly noted due to the linkage of these two mutations. Again, high Hb F with normal level of Hb A2 and other hematological parameters , CCAAT displacement protein (CDP), and an erythroid specific protein NFE132. It has been shown that the Greek/Black/Sardinian \u2212\u00a0117 A\u03b3 (G>A) and Japanese \u2212\u00a0114 G\u03b3 (C>T) HPFH mutations slightly increased the binding of CP1 and CDP but reduced the binding of NFE1 to the distal CCAAT motif32. A novel mutation at the same region, [\u2212\u00a0117 A\u03b3 (G>C)] identified in Thai subject with Hb E heterozygote in this study was associated with 13.5% Hb F. It is conceivable that this Thai \u2212\u00a0117 A\u03b3 (G>C) HPFH mutation should behave similar mechanism with that of the Greek/Black/Sardinian \u2212\u00a0117 A\u03b3 (G>A) HPFH mutation. Prediction of transcription factors binding to the region using the TFBIND program18 in comparison between the wild-type promoter, \u2212\u00a0117 A\u03b3 (G>A) Greek/Black/Sardinian HPFH, \u2212\u00a0117 A\u03b3 (G>C) Thai HPFH and \u2212\u00a0114 G\u03b3 (C>T) Japanese HPFH was carried out as shown in Table RRCCAATSA) for the CCAAT box as shown in Table A\u03b3 (G>A) with score of 0.922056. The Thai \u2212\u00a0117 A\u03b3 (G>C) and the Japanese \u2212\u00a0114 G\u03b3 (C>T) had decreased scores of 0.824343 and 0.797797, respectively.Unlike other globin genes which contain only one CCAAT motif, \u03b3-globin gene has duplicated CCAAT sequences. The proximal CCAAT motif is located at -88 nucleotide and the distal one is found at \u2212\u00a0115 regions. The proximal CCAAT motif is corresponding to the CCAAT motif of \u03b2-globin gene in which mutations in this motif result in reduced \u03b2-globin gene expression and \u03b2-thalassemia pursue. In contrast, mutations in the distal CCAAT motif of \u03b3-globin gene results in higher \u03b3-globin expression and HPFH phenotypeG\u03b3-globin gene has been proposed as a negative regulatory element based on study in transiently transfected K562 cell33. The Iranian \u2212\u00a0567 G\u03b3 (T>G) HPFH mutation (HBG2:c.\u2212\u00a0620\u00a0T>G) which changed a GATA-1 binding motif to GAGA sequence (AGATAA>AGAGAA) was associated with increased Hb F of 5.9% and 10.2% in two Iranian subjects33. The five bp deletion between \u2212\u00a0533 to \u2212\u00a0529 (-ATAAG) of G\u03b3 globin gene (HBG2:c.\u2212\u00a0582_\u2212\u00a0586del ATAAG), located at the GATA-1 binding site was also associated with HPFH phenotype in an Indian family34. Therefore, alteration of GATA-1 and related repressor proteins bindings to the motif should result in up-regulation of \u03b3-globin gene expression. In contrast, the Thai \u2212\u00a0530 G\u03b3 (A>G) HPFH mutation which is located at the same region of GATA-1 binding motif (AGATAA>AGATAG) may have less effect on G\u03b3-globin gene expression since it does not modify the GATA-1 binding site (A/T)GATA(A/G) at this position significantly. Therefore, as shown in Table silico analysis of GATA-1 related transcription factors binding of the Thai \u2212\u00a0530 G\u03b3-HPFH using the online TFBIND program showed less changes on the similarity scores as compared to the wild-type sequence which contrasts with the \u2212\u00a0533 to \u2212\u00a0529 (-ATAAG) Indian HPFH mutation which reduced binding of all GATA-1 isoforms. As shown in the table, we found that the Thai HPFH mutation had slight increased binding scores of two GATA-1 isoforms and decreased binding scores for another two GATA-1 isoforms. The mechanisms underlying up-regulation of \u03b3-globin gene in these HPFH mutations may be difference. Accordingly, the effect of the \u2212\u00a0530 G\u03b3 (A>G) Thai HPFH mutation on the G\u03b3-globin gene expression could be minimal. This might explain the marginal elevation of Hb F (1.9% and 3.0%) observed in pure heterozygotic form of the mother and the sister of the proband and indicated that two copies of the \u2212\u00a0530 G\u03b3 (A>G) mutation in homozygote state is required for dramatically increased in Hb F (11.5%) as seen in the proband and \u2212\u00a0158 XmnI G\u03b3 (\u2212) had normal Hb F level (0.3%) due to a co-inheritance of \u03b1+-thalassemia. It is evidenced from the Indian and Thai HPFH families that subjects with combined non-deletional HPFH mutation and \u03b2-thalassemia mutation in trans had elevated Hb A2 for \u03b2-thalassemia trait i.e., 3.6\u20133.9% in Indians34 and 4.3% in Thai (Table 25.Nucleotides between \u2212\u00a0675 to \u2212\u00a0526 of 0-thalassemia, A\u03b3\u03b4\u03b20-thalassemia, and HPFH described before10. Complex interactions between these non-deletional HPFH in both heterozygote and homozygote with other hemoglobinopathies commonly found in the region can lead to various hematological phenotypes and Hb F productions which could render diagnosis difficult. Identification of these non-deletional HPFH using rapid PCR diagnostic assays developed should improve this diagnosis in routine practice in the regions.In summary, three known and two novel HPFH mutations were identified for the first time in Thai population. This result indicates a diverse molecular heterogeneity of non-deletional HPFH in Thai population in addition to the deletional forms of \u03b4\u03b2Supplementary Information."} +{"text": "These chains are linked to each other by N\u2014H\u22efN hydrogen bonds, N\u2013H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions, forming a three-dimensional network.In the crystal, the mol\u00adecules are connected by N\u2014H\u22efN hydrogen bonds into dimers with an 19H15N5S, the thio\u00adphene ring is disordered in a 0.6:0.4 ratio by an approximate 180\u00b0 rotation of the ring around the C\u2014C bond linking it to the pyridine ring. In the crystal, the mol\u00adecules are linked by N\u2014H\u22efN hydrogen bonds into dimers with an R22(12) motif, forming chains along the b-axis direction. These chains are connected to each other by further N\u2014H\u22efN hydrogen bonds, forming a three-dimensional network. Furthermore, N\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 [centroid\u2013centroid separations = 3.899\u2005(8) and 3.7938\u2005(12)\u2005\u00c5] inter\u00adactions also contribute to the crystal cohesion. A Hirshfeld surface analysis indicated that the most important contributions to the surface contacts are from H\u22efH (46.1%), N\u22efH/H\u22efN (20.4%) and C\u22efH/H\u22efC (17.4%) inter\u00adactions.In the title compound, C The dihedral angle between the phenyl (C7\u2013C12) and pyridine (N1/C2\u2013C6) rings is 64.42\u2005(11) \u00b0.The pyridine ring (N1/C2\u2013C6) of the title compound Fig.\u00a01 is large3.et al., 1995b-axis direction. These chains are connected to each other by further N\u2014H\u22efN hydrogen bonds, forming a three-dimensional network (Tables 1Cg1\u22efCg1i = 3.899\u2005(8)\u2005\u00c5; slippage = 1.899\u2005\u00c5; Cg3\u22efCg3ii = 3.7938\u2005(12)\u2005\u00c5; slippage = 1.383\u2005\u00c5; symmetry codes: (i) \u2212x, 1\u00a0\u2212\u00a0y, z; (ii) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, z; Cg1 and Cg3 are the centroids of the major component of the disordered thio\u00adphene ring and of the pyridine ring, respectively] also contribute to crystal cohesion . Fig.\u00a08u. Fig.\u00a07a,b. Fig4.et al., 2016et al., 2019et al., 2014aet al., 2014bet al., 2014cThe four related compounds found as a result of the search for \u20182,6-di\u00adamino-4-(thio\u00adphen-2-yl)pyridine-3,5-dicarbo\u00adnitrile\u2019 in the Cambridge Structure Database , chains running along the b-axis direction are formed through N\u2014H\u22efO inter\u00adactions between the 1,4-di\u00adhydro\u00adpyridine N atom and one of the O atoms of the ester groups. Neighbouring chains are linked by C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. In the crystal of WOJCIJ (space group P21/c), inversion dimers linked by pairs of N\u2014H\u22efN hydrogen bonds generate P21/n), inversion dimers linked by pairs of N\u2014H\u22efNc (c = cyanide) hydrogen bonds generate Pbca), inversion dimers linked by pairs of N\u2014H\u22efNn (n = nitrile) hydrogen bonds generate In the crystal of MUCLAA (space group 5.To a solution of 2-(thio\u00adphen-2-yl\u00admethyl\u00adene)malono\u00adnitrile and malono\u00adnitrile in methanol (25\u2005mL), phenyl\u00adethyl\u00adamine was added and the mixture was stirred at room temperature for 48\u2005h. Then 15\u2005mL of methanol were removed from the reaction mixture, which was left overnight. The precipitated crystals were separated by filtration and recrystallized from ethanol/water (1:1) solution .1H NMR : 1.55 ; 5.45 ; 7.21\u20137.88 ; 13C NMR : 21.69 (CH3), 50.00 (CH\u2014Ar), 79.77 (=Ctert), 80.92 (=Ctert), 116.85 (CN), 116.97 (CN), 127.14 (2CHarom), 127.22 (CHarom), 128.11 (CHthien\u00adyl), 128.63 (2CHarom), 130.14 (CHthien\u00adyl), 130.75 (CHthien\u00adyl), 134.53 (Car), 144.53 (Cthien\u00adyl), 152.30 (=Ctert), 158.70 (N=Ctert), 161.38 (=Ctert).6.SHELXL were used for the Uij values of equivalent atom pairs and DFIX commands were used to restrain the nearest-neighbour and next-nearest-neighbour bond distances in the two disorder components to be equal with a standard deviation of 0.03\u2005\u00c5. All C-bound H atoms were placed in calculated positions (0.95\u20131.00\u2005\u00c5) and refined as riding with Uiso(H) = 1.2 or 1.5Ueq(C). The N-bound H atoms were located in a difference map and refined with Uiso(H) = 1.2Ueq(N) .Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023003845/vm2282sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989023003845/vm2282Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023003845/vm2282Isup3.cmlSupporting information file. DOI: 2260011CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "These ribbons are connected by N\u2014H\u22efO, N\u2014H\u22efN hydrogen bonds and van der Waals inter\u00adactions.In the crystal, mol\u00adecules are linked by inter\u00admolecular N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds into ribbons parallel to (022) along the 11H7N5OS, contains two independent mol\u00adecules (1 and 2). The thio\u00adphene ring in mol\u00adecule 2 is rotationally disordered (flip disorder) by ca 180\u00b0 over two sites with the site-occupation factors of 0.9 and 0.1. These two orientations of the thio\u00adphene ring in mol\u00adecule 2 are not equivalent. In the crystal, mol\u00adecules are linked by inter\u00admolecular N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds into ribbons parallel to (022) along the a axis. Within the (022) planes, these ribbons are connected by van der Waals inter\u00adactions and between the (022) planes by N\u2014H\u22efO hydrogen bonds. In mol\u00adecule 1, Hirshfeld surface analysis showed that the most important contributions to the crystal packing are from N\u22efH/H\u22efN (27.1%), H\u22efH (17.6%), C\u22efH/H\u22efC (13.6%) and O\u22efH/H\u22efO (9.3%) inter\u00adactions, while in mol\u00adecule 2, H\u22efH (25.4%) inter\u00adactions are the most significant contributors to the crystal packing.The asymmetric unit of the title compound, C These two orientations of the thio\u00adphene ring in mol\u00adecule 2 are not equivalent.As seen in Fig.\u00a01Database survey section.In mol\u00adecule 1, the angle between the thio\u00adphene (S1/C8\u2013C11) and pyridine (N1/C2\u2013C6) rings is 50.72\u2005(9)\u00b0. In mol\u00adecule 2, the angle between the two disordered thio\u00adphene rings (S2/C19\u2013C22 and S2\u2032/C19/C20\u2032\u2013C22\u2032) is 6.2\u2005(5)\u00b0, and they make an angle with the pyridine ring (N6/C13-C17) of 40.3\u2005(1) and 34.8\u2005(5)\u00b0, respectively. Mol\u00adecules 1 and 2 (r.m.s. deviation = 0.126\u2005A) are almost identical and the geometric parameters are normal and comparable to those of related compounds listed in the B\u22efN2 and N10\u2014H10A\u22efN7 hydrogen bonds, forming S(5) motifs along the s Table\u00a01. Within s Table\u00a01 and 4 \u25b8.CrystalExplorer17.5 represent N\u2014H\u22efO inter\u00adaction zones (Table\u00a01b) inter\u00adactions provide the highest contribution (Table\u00a02c), C\u22efH/H\u22efC and O\u22efH/H\u22efO contacts are also significant. Table\u00a02dnorm mappings were performed in the range \u22120.5165 to +1.1535 a.u. The locations of N\u2014H\u22efO inter\u00adactions are shown by bright-red circles on the dnorm surfaces , Table\u00a01c; 25.4%) are the major factor in the crystal packing with N\u22efH/H\u22efN , O\u22efH/H\u22efO and C\u22efH/H\u22efC inter\u00adactions representing the next highest contributions. The percentage contributions of comparatively weaker inter\u00adactions in mol\u00adecules 1 and 2 are given in Table\u00a02es Fig.\u00a05a,b reprs Table\u00a01. The fins Table\u00a01 show thas Table\u00a01b inter\u00adn Table\u00a02, as wouln Table\u00a02c, C\u22efH/Hes Fig.\u00a05c,d, Tab\u22efN Fig.\u00a06b; 24.3%\u22efO Fig.\u00a06e; 11.7%\u22efC Fig.\u00a06d; 11.4%4.et al., 2016viz. CSD refcodes BEFFOL , pairs of mol\u00adecules are linked by pairs of N\u2014H\u22efN hydrogen bonds, forming dimers with an II crystallizes in the triclinic space group PIIA and IIB) in the asymmetric unit. In the crystal of II, mol\u00adecules IIA and IIB are linked by inter\u00admolecular N\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds into layers parallel to (001). These layers are connected along the c-axis direction by weak C\u2014H\u22efN contacts. C\u2014H\u22ef\u03c0 and C\u2014N\u22ef\u03c0 inter\u00adactions connect adjacent mol\u00adecules, forming chains along the a-axis direction. In III (space group: Pc), the two mol\u00adecules in the asymmetric unit are joined together by N\u2014H\u22efO hydrogen bonds, forming a dimer with an c-axis direction, which are connected by C\u2014Br\u22ef\u03c0 inter\u00adactions. In IV (space group: Pc), inter\u00admolecular N\u2014H\u22efN and C\u2014H\u22efN hydrogen bonds, as well as N\u2014H\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions, connect the mol\u00adecules in the crystal, generating a 3D network. In both V (space group: PVI (space group: PVII (space group: P21/n), the crystal structure is stabilized by inter\u00admolecular C\u2014H\u22efF and C\u2014H\u22ef\u03c0 inter\u00adactions, and in VIII (space group: P21/c), by inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. In IX (space group: Cc), inter\u00admolecular N\u2014H\u22efN and C\u2014H\u22efN hydrogen bonds form mol\u00adecular sheets parallel to the (110) and (110) planes, crossing each other. Adjacent mol\u00adecules are further linked by C\u2014H\u22ef\u03c0 inter\u00adactions, which form zigzag chains propagating parallel to [100]. The compound X (space group: Pca21) crystallizes with two independent mol\u00adecules in the asymmetric unit. In the crystal, the A and B mol\u00adecules are linked by N\u2014H\u22efS, N\u2014H\u22efN and C\u2014H\u22efS hydrogen bonds, forming a three-dimensional network. In XI (space group: P21/c), mol\u00adecules are linked into a chain along the b-axis direction via C\u2014H\u22efO inter\u00adactions. In XII (space group: PXIII (space group: P21/c), the mol\u00adecules form centrosymmetric dimers via N\u2014H\u22efS hydrogen bonds.In the crystal of 5.et al., 2020The title compound was synthesized using a recently reported procedure (Babaee 6.Uiso(H) = 1.2Ueq(C). The N-bound H atoms were found in a difference-Fourier map, and refined freely , with Uiso(H) = 1.2Ueq(N). The thio\u00adphene ring (S2/C19\u2013C22) in mol\u00adecule 2 is rotationally disordered (flip disorder) by ca 180\u00b0 over two sites with the site-occupation factors of 0.9 and 0.1 (fixed after refinement cycles). A DFIX instruction was applied to constrain the distances in the thio\u00adphene rings of disordered mol\u00adecule 2. For these rings, FLAT and EADP instructions were also used.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023003237/tx2066sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023003237/tx2066Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023003237/tx2066Isup3.cmlSupporting information file. DOI: 2255359CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A co-crystal of quabodepistat and 2,5-di\u00adhydroxy\u00adbenzoic acid was obtained and the crystal structure was solved from microcrystal electron diffraction (MicroED) data. R,4R)-1--3,4-di\u00adhydroxy\u00adpiperidin-4-yl]meth\u00adoxy}-8-fluoro-3,4-di\u00adhydro\u00adquinolin-2(1H)-one); C21H20ClF3N2O4] and 2,5-di\u00adhydroxy\u00adbenzoic acid were successfully co-crystallized. Given the small size of the crystals (1 \u00d7 0.2 \u00d7 0.2\u2005\u00b5m) the structure was solved via microcrystal electron diffraction (MicroED). The C\u2014O and C=O bond-length ratio of the carb\u00adoxy\u00adlic group in 2,5DHBA is 1.08 (1.34\u2005\u00c5/1.24\u2005\u00c5), suggesting that 2,5DHBA remains protonated. Therefore, the material is a co-crystal rather than a salt. The amide group of quabodepistat participates in a cyclic hydrogen bond with the carb\u00adoxy\u00adlic group of the 2,5DHBA. Additional hydrogen bonds involving the quabodepistat amide and hydroxyl groups result in a three-dimensional network.Quabodepistat [(5-{[(3 The angles O21\u2014C16\u2014C17, F30\u2014C28\u2014C23, C6\u2014N7\u2014C8, C9\u2014C10\u2014C11, O1\u2014C14\u2014C15, and C6\u2014C11\u2014C2 also have z-score values greater than 3.Quabodepistat and 2,5DHBA co-crystallize in a 1:1 stoichiometric ratio in the monoclinic system, space group 2 values exhibit \u03c73.via hydrogen bonds are observed between quabodepistat and 2,5DHBA. One of the inter\u00adactions is between a carb\u00adoxy\u00adlic group and an amide. As shown in Fig.\u00a01et al., 2020Inter\u00admolecular inter\u00adactions 4.et al., 2016A search for co-crystals with 2,5-di\u00adhydroxy\u00adbenzoic acid (or gentisic acid) in the Cambridge Structural Database . Tetra\u00adhydro\u00adfuran (THF) and hexane were purchased from FUJIFILM Wako Pure Chemical Corporation . 2,5DHBA was purchased from Tokyo Kasei Kogyo Co., Ltd. . Quabodepistat (5\u2005g) and 2,5DHBA were dissolved in 100\u2005mL of THF. 250\u2005mL of hexane were added while stirring. Precipitation occurred as soon as hexane was added. The THF/hexane was stirred at room temperature (approximately 298\u2005K) for three days. After filtration, it was dried at room temperature for 24\u2005h, then heated at 383\u2005K for 20\u2005h.6.\u2212\u00c5\u22122 s\u22121. Contiguous diffraction frames were collected every 0.5\u00b0 from each crystal by continuously rotating the sample stage at a goniometer rotation speed of 1\u00b0 s\u22121; the sample stage was rotated from \u221240\u00b0 to 40\u00b0 for the first crystal and from \u221260\u00b0 to 60\u00b0 for the second crystal . The structure was refined kinematically. Refinement with SHELXL was carried out using the scattering factors for electron diffraction global, I. DOI: Click here for additional data file.10.1107/S2056989023006047/dj2052Isup2.cmlSupporting information file. DOI: https://doi.org/10.5281/zenodo.7156704Deposit the raw data into Zenodo: https://dx.doi.org/10.5517/ccdc.csd.cc2d19zrDeposit the raw data into CCDC: 2205804CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the title compound consists of two independent mol\u00adecules differing slightly in conformation and in their inter\u00admolecular inter\u00adactions in the solid. 8H7NO3, consists of two mol\u00adecules differing slightly in conformation and in their inter\u00admolecular inter\u00adactions in the solid. The dihedral angle between the benzene and dioxolane rings is 0.20\u2005(7)\u00b0 in one mol\u00adecule and 0.31\u2005(7)\u00b0 in the other. In the crystal, the two mol\u00adecules are linked into dimers through pairwise O\u2014H\u22efN hydrogen bonds, with these units being formed into stacks by two different sets of aromatic \u03c0-stacking inter\u00adactions. The stacks are connected by C\u2014H\u22efO hydrogen bonds. A Hirshfeld surface analysis indicates that the most significant contacts in the crystal packing are H\u22efO/O\u22efH (36.7%), H\u22efH (32.2%) and C\u22efH/H\u22efC (12.7%).The asymmetric unit of the title mol\u00adecule, C For the C1\u2013C6 rings, the centroid\u2013centroid distance is 3.6024\u2005(11)\u2005\u00c5 with a slippage of 1.185\u2005\u00c5 between mol\u00adecules at x, y, z and \u2212x, \u2212y\u00a0+\u00a01, \u2212z. These paired mol\u00adecules make weak, slipped \u03c0-stacking inter\u00adactions with corresponding pairs at \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01 with a centroid\u2013centroid distance of 3.8479\u2005(11)\u2005\u00c5 and a slippage of 1.947\u2005\u00c5. The C9\u2013C14 ring has slipped \u03c0-stacking inter\u00adactions with its counterparts in mol\u00adecules at x\u00a0\u2212\u00a0y\u00a0+\u00a0z\u00a0\u2212\u00a0x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a0B\u22efO4, C8\u2014H8\u22efO6, C15\u2014H15A\u22efO1 and C16\u2014H16\u22efO3 hydrogen bonds -2-(1H-imidazol-1-yl)ethyl\u00adidene]hydroxyl\u00adamine N-hy\u00addroxy-3-(1H-imidazol-1-yl)propan-1-imine iso\u00adpropanol solvate form and (Z)-3,4-methyl\u00adene\u00addioxy\u00adbenzaldehyde oximium 4-toluene\u00adsulfonate A search using CCDC ConQuest of the Cambridge Structural Database and 5(b), respectively. Fig.\u00a06a)] and delineated into C\u22efH/H\u22efC [Fig.\u00a06c)], and H\u22efO/O\u22efH [Fig.\u00a06h)] inter\u00adactions. For completeness, the H\u22efH inter\u00adactions constitute 32.2% of the surface [Fig.\u00a06b)]. The other inter\u00adactions contribute small amounts, viz., C\u22efN/N\u22efC (1.0%), C\u22efO/O\u22efC (2.4%), C\u22efC (9.5%), H\u22efN/N\u22efH (4.1%), N\u22efO/O\u22efN (1.1%), N\u22efN (0.0%) and O\u22efO (0.4%).The Hirshfeld surface analysis was performed with 6.d]dioxole-5-carbaldehyde dissolved in 50\u2005ml of ethanol was added to the mixture. After 5\u2005h of stirring at 273\u2005K, the product was allowed to precipitate and then filtered with a yield of 90%. Single crystals were recrystallized from ethanol solution.A solution of 5.0\u2005g of sodium hydroxide dissolved in 20\u2005ml of water was mixed with 8.0\u2005g of hydroxyl\u00adamine hydro\u00adchloride dissolved in 15\u2005ml of water, then 8.0\u2005g of benzo[7.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989023004139/hb8064sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023004139/hb8064Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023004139/hb8064Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023004139/hb8064Isup4.cmlSupporting information file. DOI: 2262070CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E)-1-[(carbamo\u00adthioyl\u00adamino)\u00adimino]\u00adeth\u00adyl}-4-methyl-1,3-thia\u00adzol-3-ium cations are linked by O\u2014H\u22efCl, N\u2014H\u22efCl, N\u2014H\u22efO, N\u2014H\u22efS and C\u2014H\u22efS hydrogen bonds, forming a tri-periodic network.In the crystal, the 2-amino-5-{-1-[(carbamo\u00adthioyl\u00adamino)\u00adimino]\u00adeth\u00adyl}-4-methyl-1,3-thia\u00adzol-3-ium cation, one chloride anion and one water mol\u00adecule of crystallization. The cation is nearly flat (r.m.s. deviation of non-H atoms is 0.0814\u2005\u00c5), with the largest deviation of 0.1484\u2005(14)\u2005\u00c5 observed for one of the methyl C atoms. In the crystal, the cations are linked by O\u2014H\u22efCl, N\u2014H\u22efCl, N\u2014H\u22efO, N\u2014H\u22efS and C\u2014H\u22efS hydrogen bonds, forming a tri-periodic network. The most important contributions to the crystal packing are from H\u22efH (35.4%), S\u22efH/H\u22efS (24.4%), N\u22efH/H\u22efN (8.7%), Cl\u22efH/H\u22efCl (8.2%) and C\u22efH/H\u22efC (7.7%) inter\u00adactions.In the hydrated title salt, C In the 1,3-thia\u00adzol-3-ium ring, as expected, the C1\u2014N2 distance of 1.3309\u2005(16)\u2005\u00c5 indicates double-bond character, while the C2\u2014N2 distance of 1.3885\u2005(14)\u2005\u00c5 has more single-bond character.The asymmetric unit of (I)N\u2032-[(1Z)-ethyl\u00adidene]ethane\u00adthio\u00adhydrazide group, the S2\u2014C7\u2014N4\u2014N3, N5\u2014C7\u2014N4\u2014N3, C7\u2014N4\u2014N3\u2014C5 and N4\u2014N3\u2014C5\u2014C6 torsion angles are 178.17\u2005(8), \u22120.63\u2005(16), 174.48\u2005(10) and 0.16\u2005(18)\u00b0, respectively. The title compound shows bond lengths and angles that are typical and are in agreement with those reported for the related compounds discussed in the Database survey section.In the amino-The cation is nearly flat (r.m.s. deviation of the 14 non-H atoms is 0.0814\u2005\u00c5), with the largest deviations observed for C6 [0.1484\u2005(14)\u2005\u00c5], N1 [0.1357\u2005(10)\u2005\u00c5], and S2 [0.1399\u2005(6)\u2005\u00c5].3.In the crystal of (I)Crystal Explorer 17.5 , ds Fig.\u00a06.The two-dimensional fingerprint plots of the most abundant contacts are presented in Fig.\u00a074.et al., 2016et al., 2012et al., 2003et al., 1996et al., 2011A search of the Cambridge Crystallographic Database , the mol\u00adecular packing is determined by inter\u00adionic N\u2014H\u22efCl contacts. In the crystal of (V), the mol\u00adecules of substituted urea are connected by O\u2014H\u22efO hydrogen bonds into sheets. In turn, these sheets are connected to each other via N\u2014H\u22efO hydrogen bonds with hydrogen phosphate anions, forming a tri-periodic network.In the crystal of (II), mol\u00adecules are linked by O\u2014H\u22efCl and N\u2014H\u22efCl hydrogen bonds, forming zigzag chains along [001]. There is also a C\u2014H\u22efCl inter\u00adaction present. The crystal structure of (III) comprises a substituted thia\u00adzolium ring that is connected to a nitrate ion 5.et al., 2016v:v) solution at room temperatureThe title compound was synthesized using a reported procedure (Gomha 6.Uiso(H) = 1.5Ueq(C). The H atoms attached to the N atom and the H atoms of the water mol\u00adecule were found in a difference-Fourier map. Their positional parameters were refined freely while setting Uiso(H) = 1.2Ueq(N) and 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023007090/wm5691sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023007090/wm5691Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023007090/wm5691Isup3.cmlSupporting information file. DOI: 2288159CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Single-crystal X-ray crystallographic analysis of 1 confirmed the presence of the first known (arene)cuprate. Also, unlike all previously known homoleptic (anthracene)metallates of d-block elements, which con\u00adtain metals coordinated only to terminal rings, the organocuprate unit in 1 con\u00adtains copper bound to the 9,10-carbons of the central ring of anthracene. No other d- or f-block metal is known to afford an anthracene or other aromatic hydro\u00adcarbon com\u00adplex having the architecture of organodicuprate 1.Reactions of (tri\u00adcyclo\u00adhexyl\u00adphosphane)copper(I) chloride with two equivalents of potassium anthracene (KAn) in tetra\u00adhydro\u00adfuran (THF) at 200\u2005K provides air-sensitive but thermally stable (at 293\u2005K) solutions from which yellow crystalline blocks of bis\u00ad[bis\u00ad(tetra\u00adhydro\u00adfuran-\u03ba Although the cuprate was originally speculated to be a copper analog of the mono\u00adatomic gold anion, Au1\u2212, established to be present in cesium auride, CsAu , which have been proposed to exist in solid neon at 4\u201310\u2005K \u00admetallates , in THF. Also, because stoichiometrically analogous naph\u00adtha\u00adlene and anthracene com\u00adplexes of a given d-block element may possess very similar mol\u00adecular structures, particularly in the solid state ]n ]2 (Cy\u00a0= cyclo\u00adhex\u00adyl) adducts of copper(I) halides due to the ease with which coordinated or free PPh3 and mixed tertiary ar\u00adyl\u2013alkyl phosphanes undergo reductive cleavage of P\u2014C(ar\u00adyl) bonds, unlike tri\u00adalkyl\u00adphosphanes ]2 with four equivalents of MAn in THF appear by NMR spectra to afford similar products in solution, owing to the facile isolation and crystallization of the potassium salt, only the latter will be described now. Thus, the addition of a colorless solution of [CuCl(PCy3)]2 in THF to a dark-blue solution of KAn (molar ratio: 2 KAn/Cu) in THF at 200\u2005K, led to the formation of an air-sensitive but thermally stable (at 293\u2005K) yellow\u2013brown solution, from which yellow crystalline blocks of [K(THF)2]2[{Cu}2], 1, were isolated halides, 2\u2212 , with X\u00a0= Cl \u2005\u00c5] is long to 3.98\u2005(1)\u2005\u00c5.To the best of our knowledge, only the long-known dimeric organoalanate com\u00adplexes is well within the range of 1.83\u20132.05\u2005\u00c5 observed previously for homoleptic diorganocuprates \u2005\u00c5. Prior studies suggest that d10Cu\u2013d10Cu inter\u00adactions are very weak to non-existent when the Cu\u22efCu separation is greater than \u223c2.50\u2005\u00c5 in mol\u00adecular entities con\u00adtaining CuI com\u00adplexes dicuprate(2\u2212) com\u00adplex, relative to unknown monomeric monoanions, i.e. [{Cu(THF)x}]1\u2212 (x\u00a0= 0\u20132), which would likely be strained metallacycles, vide infra. In the case of the structurally related organoaluminium com\u00adplexes 2 and 3, it was proposed that the dimeric, rather than plausible monomeric structures, \u2018are favored by smaller deviations from tetra\u00adhedral angles about the aluminum atoms in the dianions rather than about the aluminum atom in the hypothetical monoanions.\u2019 However, surprisingly, the possible importance of contact ion-pairing in contributing to the stabilization of the dimers in the solid state, and possibly also in solution, was not considered ]1\u2212 (Np\u00a0= naph\u00adtha\u00adlene), based only on a proton NMR spectrum in THF and identification of 1,4-di\u00adhydro\u00adnaph\u00adtha\u00adlene as a key hydrolysis product anthracene)], 4, strongly suggests that the natures of 2 and 3 in solution merit reinvestigation. It should also be emphasized that presently we cannot rule out the possibility that monomeric forms of 1 could be present in THF or other solvents distance of 2.057\u2005(4)\u2005\u00c5, which is significantly longer than the normal Al\u2014C(Et) distance of 1.966\u2005(4)\u2005\u00c5 present in 4 ], 5, possessing a C9\u2014Mg\u2014C10 angle of 71.4\u00b0. This angle is significantly sharper than the corresponding angle in 4, likely owing in part to the higher coordination number of Mg in 5. The latter also con\u00adtains a rather long Mg\u2014C distance of 2.30\u2005(2)\u2005\u00c5 \u2005\u00c5 metal com\u00adplex to be recognized as a strained metallacycle ], 6, or 2,3:5,6-dibenzo-7-di\u00admethyl\u00adger\u00admano\u00adrbor\u00adna\u00addiene. X-ray structure characterization of 6, Me2GeAn, revealed a sharp C9\u2014Ge\u2014C10 angle of 77.72\u2005(5)\u00b0, and Ge\u2014C distances [average 2.030\u2005(1)\u2005\u00c5] which are significantly longer than the Ge\u2014C(Me) distance [average 1.943\u2005(4)\u2005\u00c5] \u2005\u00c5; Hencher & Mustoe, 19756 suffers thermal loss of anthracene at 373\u2005K in toluene to produce in good yield an intriguing di\u00admethyl\u00adgermylene, Me2Ge, adduct [{Ge2Me4}], 7, which may arise via insertion of highly reactive Me2Ge into a strained Ge\u2014C(bridgehead) bond of 6. However, also possible is an initial dimerization of Me2Ge to the digermene, Me4Ge2, followed by its facile [4\u00a0+\u00a02] cyclo\u00adaddition to free anthracene, which was established previously to afford 7 of ca \u221237\u2005kcal\u2005mol\u22121 of 7 Mg solvate in crystalline 1\u2212, the monomer of structurally characterized dimeric 3 ]z (z\u00a0= 0 or \u22121), are presently known to form dimers (or oligomers) related to 1, 2, or 3. Calculations on the structures and relative stabilities of the monomeric forms of 1, 2, and 3, com\u00adpared to the respective dimers, would be of considerable inter\u00adest. Inclu\u00adsion of contact ion-pairing could be of key importance in such a study, but might prove to be a nontrivial extension.Of particular relevance to our discussion is that neutral 1, which con\u00adtains only Cu\u2014C \u03c3-bonds, all prior (arene)copper com\u00adpounds have been characterized as \u03c0-com\u00adplexes, including the homoleptic cationic species, [Cu(arene)2]1+ [(\u03bc-Cl)3AlCl] (PR3)]PF6 (R\u00a0= phenyl or phenoxide) were described am\u00adino\u00admeth\u00adyl)anthracene}]1+ com\u00adplex, in which CuI is con\u00adstrained via chelation with di\u00adphenyl\u00adphosphanyl groups to lie over the central ring of nearly planar anthracene and 2.414\u2005(6)\u2005\u00c5] for asymmetric \u03c0-bonding of the 1-naph\u00adthyl group, the Cu\u2014C distances in the copper\u2013\u03b76-anthracene com\u00adplex are in the range 2.773\u20133.021\u2005\u00c5. These are much longer than usual in (arene)copper com\u00adplexes, indicative, at best, of a very weak copper\u2013anthracene inter\u00adaction (PR3)]PF6, the Cu\u2014C distances are in the range 2.253\u20132.300\u2005\u00c5 ]2 (Cy\u00a0= cyclo\u00adhex\u00adyl) was prepared as described previously colorless solution of [CuCl(PCy3)]2 in THF (50\u2005ml) and stirring continued for 12\u2005h at 200\u2005K. The resulting yellow\u2013brown solution was warmed over a ca 6\u2005h period to near 290\u2005K (room tem\u00adper\u00ada\u00adture) and filtered (medium or P4 porosity frit) to remove KCl. After careful evaporation of all but about 20\u2005ml of solvent in vacuo at 273\u2013293\u2005K, pentane (100\u2005ml) was added with stirring. The resulting slurry was filtered, washed vigorously with pentane (2 \u00d7 10\u2005ml) and dried in vacuo to afford 0.59\u2005g of homogeneous yellow solid [K(THF)2]2[{Cu(C14H10)}2], 1. No elemental analysis was con\u00adducted on the bulk solid 1, so its com\u00adposition is based exclusively on the SCXRD study and NMR spectra. X-ray-quality yellow blocks of 1 were grown from a pentane-layered saturated solution in THF at 240\u2005K over a 6\u2005d period.Sublimed anthracene and shiny pieces of potassium metal were transferred in an argon-filled glove-box to a round-bottomed Schlenk flask, along with a glass-enclosed magnetic stirrer bar. Subsequently, THF (100\u2005ml) was added and the mixture was stirred vigorously in the dark for 6\u2005h at 293\u2005K to afford a deep-blue solution of potassium anthracene (KAn), which is susceptible to photo-oxidation by visible light. This solution/slurry was cooled to 200\u2005K with stirring and to it was transferred 1H NMR : 1.77 , 3.52 , 3.63 , 6.24 , 6.35 . 13C{1H} NMR : 24.2 , 52.4 , 66.3 , 117.7 , 117.9 , 145.2 .3}], 5 : 3.51 , 5.95 , 6.01 ; 13C{1H} NMR : 57.7 , 114.1 , 118.1 , 145.9 monomeric (anthracene)metal com\u00adplex, 2[{Cu}2], con\u00adtains one copper center and one anthracene ligand in contact with one [K(THF)2]1+ counter-ion. A crystallographic inversion center generates full well-resolved 1 (\u03b76-Np)]2\u2212 metal dimer com\u00adplex present in [Na(THF)2]2[{(AlMe2)}2], 3, is greater than the corresponding sum for Na and Al (2.87\u2005\u00c5) \u2005\u00c5, K1\u22efCu1A\u00a0= 3.3762\u2005(9)\u2005\u00c5, K1\u22efK1A\u00a0= 6.542\u2005(2)\u2005\u00c5 and Cu1\u22efCu1A\u00a0= 2.6171\u2005(7)\u2005\u00c5 , than the corresponding distances in 3, i.e. Na1\u22efAl1\u00a0= 4.348\u2005(6)\u2005\u00c5, Na1\u22efAl1A\u00a0= 4.455\u2005(7)\u2005\u00c5, Na1\u22efNa1A\u00a0= 7.28\u2005(1)\u2005\u00c5 and Al1\u22efAl1A\u00a0= 4.95\u2005\u00c5. As described in Section\u00a011 arises from the essentially linear C9\u2014Cu\u2014C10A angle, expected for two-coordinated d10 CuI \u00b0], resulting from the distorted tetra\u00adhedral geometry of four-coordinated AlIII, causes the Al\u22efAl distance to be much longer \u2005\u00c5] and, as a result, its mol\u00adecular structure, without the THF groups, is fairly symmetrical and deviates only slightly from Dh2 symmetry \u2005\u00c5 and 3.57\u2005(1)\u2005\u00c5, respectively], which show that the two formal 9,10-coordinated anthracene dianion units are slightly further apart in 1 than they are in 3, owing also to the different coordination numbers of copper and aluminium in these remarkable com\u00adpounds, both of which are worthy of additional study.However, in er Fig.\u00a02. Of part\u2005\u00c5 Fig.\u00a01, with on1 and 3 are similar. Thus, for 1, the bridgehead angles C11\u2014C10\u2014C14 [113.3\u2005(2)\u00b0] and C12\u2014C9\u2014C13 [113.1\u2005(2)\u00b0] (Table\u00a023 , which are both close to the value of 1.51\u2005(5)\u2005\u00c5 observed previously for C(sp2)\u2014C(sp3) distances 2}]1\u2212, is structurally nearly identical, without the metal, to those observed in the dianions of 1 and 3, with an average bridgehead C\u2014C distance of 1.485\u2005(9)\u2005\u00c5. The lutetium anion also has an average bridgehead C\u2014C\u2014C angle of 111.8\u2005(5)\u2005\u00c5, which is very close to the corresponding angles of 1 and 3. A key structural difference in the lutetium anion is the sharp C9\u2014Lu\u2014C10 angle of 67.1\u2005(2)\u00b0, owing to the metalla\u00adcyclo\u00adpropane character of the com\u00adplex and, likely also the large bulk of the bis\u00ad(cyclo\u00adpenta\u00addien\u00adyl)lutetium moiety, which may well be responsible for its stability towards dimerization or oligomerization in solution (THF) and in the solid state as a crystalline [Na(diglyme)2]1+ salt (diglyme\u00a0= di\u00adethyl\u00adene glycol dimethyl ether) at ca 293\u2005K having the shortest distances [average 3.14\u2005(13)\u2005\u00c5], whereas the K\u22efC distances for the six C atoms closest to the long K\u22efCu contact are longer [average 3.38\u2005(5)\u2005\u00c5] to 3.24\u2005(1)\u2005\u00c5, but the Na\u2014C distances of the four more crowded ring-junction C atoms are mostly longer and C\u2014C\u2014C angles [average 119.9\u2005(2)\u00b0] that are quite similar to those found in uncharged free benzene dicuprate(2\u2212) with relatively weakly inter\u00adacting cations, such as [K([2.2.2.]cryptand)]1+, tetra\u00adalkyl\u00adammonium\u00ad(1+), etc., would be of considerable inter\u00adest. As our laboratory will be irreversibly shuttered in 2023, others are encouraged to examine these possibilities and related issues, vide infra.Inter\u00adactions of the alkali metal cations with the 4-naph\u00adtha\u00adlene)\u00adzir\u00adco\u00adnate(2\u2212), the first confirmed homoleptic (polyarene)metallate of a d-block metal 6]2\u2212, our research group has been inter\u00adested in discovering what other d-block elements, throughout the periodic table, would afford related and hopefully labile com\u00adplexes. Thus began our exploration in a systematic fashion of the synthesis and reactivity patterns of transition-metal com\u00adpounds con\u00adtaining metals in formal negative oxidation states 3, and particularly organic isocyanides, to determine whether the anthracenes would be displaced to produce new formal Cu com\u00adplexes. Noteworthy is that although isolable Cu1\u2212 com\u00adplexes remain unknown, very recently, the first unambiguous Cu0 com\u00adplex was isolated, thoroughly characterized, and structurally authenticated I, global. DOI: 10.1107/S2053229623008367/eq3012Isup2.hklStructure factors: contains datablock(s) I. DOI: 2296812CCDC reference:"} +{"text": "N-butylamine (DBM). The aromatic compounds chosen were 2-aminopyridine (A2MP), 2-amino-4-methylpyridine (A24MP), 2-amino-6-methylpyridine (A26MP) and 4-dimethylaminopyridine (DMAP). The salts were characterised using single-crystal X-ray diffraction, thermal analysis, FTIR spectroscopy and Hirshfeld surface analysis. For all the crystal structures, N-H\u00b7\u00b7\u00b7O and C-H\u00b7\u00b7\u00b7Cl contacts were present. O-H\u00b7\u00b7\u00b7O contacts were found in all the crystal structures except for (3-chloro-4-hydroxyphenylacetic acid (CHPAA) is a fungal metabolite. It is a small molecule that is useful in crystal engineering studies due to the functional groups present. Six amines were selected to form salts with CHPAA. Linear derivatives included diethylamine (DEA) and di- N-butylamine (DBM), 2-aminopyridine (A2MP), 2-amino-4-methylpyridine (A24MP), 2-amino-6-methylpyridine (A26MP) and 4,4-dimethylaminopyridine (DMAP). These are depicted in Ka [Ka values of the amines are DEA (10.58), DBM (10.75), A2MP (6.84), A24MP (7.62), A26MP (7.60) DMAP (8.78). Thus, the \u2206pKa values ranged from 3.78 to 7.69. Salt formation is expected when \u2206pKa > 4. If <\u22121 \u2206pKa < 4, then either salt or co-crystal can form [Multicomponent crystals are important in the pharmaceutical industry as the formation of new solid forms can improve the physicochemical properties of active pharmaceutical ingredients ,2. The ced in Ka of CHPAAcan form . In thisP\u012a, Z = 2 (d(C11\u00b7\u00b7\u00b7O2) of 3.4706(18) \u00c5 as well as contacts to the chlorine atom; C-H\u00b7\u00b7\u00b7Cl (d(C11\u00b7\u00b7\u00b7Cl1) = 3.821(2) \u00c5) and N-H\u00b7\u00b7\u00b7Cl (d(N1\u00b7\u00b7\u00b7Cl1) = 3.4818(16) \u00c5). The hydrogen bond metrics are summarised in 2\u2212 anions point directly towards each other, forming a type I halogen bond with d(Cl\u00b7\u00b7\u00b7Cl) = 3.517(2) \u00c5 and an angle of 174\u00b0 . Two mod(1.76 \u00c5) ,24 and W(1.76 \u00c5) . Subsequ(1.76 \u00c5) ,27,28. Tivatives . The DEAchannels c within C2/c with Z = 8 .th Z = 8 a. Both Nchannels b. The voP\u012a, Z = 2 (\u2212 anions (3.639(1) \u00c5) and between 2+AMP cations (3.835(1) \u00c5).\u012a, Z = 2 a. There am below b, the ca1/cP2 and its asymmetric unit contains one anti-oriented (H13A) on the primary amine forms an additional hydrogen bond, allowing the structure to generate an extended network system described as chains oftructure a, the acd anions b are arrP\u012a and its asymmetric unit contains two a-axis. The \u03c0-\u03c0 stacking interactions between synthons a consist diagram b shows cPbca and its asymmetric unit contains one 2 group in DMAP. There are \u03c0\u2013\u03c0 stacking interactions between adjacent b-axis.The salt of 4-dimethylaminopyridinium-3-chloro-4-hydroxyphenylacetate, ractions a. The st\u2212)(DBM+) displayed C-H\u00b7\u00b7\u00b7O interactions. The O-H\u00b7\u00b7\u00b7O and N-H\u00b7\u00b7\u00b7O hydrogen bonds gave contact distances with average values for d(H\u00b7\u00b7\u00b7O) = 1.79 \u00c5, d(H\u00b7\u00b7\u00b7N) = 1.89 \u00c5, d(O\u00b7\u00b7\u00b7O) = 2.62 \u00c5 and d(N\u00b7\u00b7\u00b7O) = 2.78 \u00c5. These were followed by the C-H\u00b7\u00b7\u00b7O and C-H\u00b7\u00b7\u00b7Cl contacts which gave average values of d(H\u00b7\u00b7\u00b7O) = 2.55 \u00c5, d(H\u00b7\u00b7\u00b7 Cl) = 3.01 \u00c5, d(C\u00b7\u00b7\u00b7O) = 3.44 \u00c5 and d(C\u00b7\u00b7\u00b7Cl) = 3.72 \u00c5.In summary, and Cl\u00b7\u00b7\u00b7H (15.8\u201318.8%) contacts. There are prominent wings on the fingerprint plots for\u2212 anions in these structures. Only . Thus, salt formation with the aromatic amines resulted in increased thermal stability with the highest melting point observed for Differential scanning calorimetry (DSC) results are presented for all the salts, and thermogravimetric analysis (TGA) was completed for , A24MP (3429 cm\u22121) and A26MP (3461 cm\u22121) either disappeared or diminished in the IR spectra of the resulting salts. This can be attributed to the \u2212NH2 group\u2019s involvement in hydrogen bonding in the A2MP, A24MP and A26MP salts. The FTIR spectra are given in FTIR spectroscopy is often used in screening new solid forms to distinguish between co-crystal and salt formation ,33. It iPXRD analyses of all compounds obtained from the grinding and slurry experiments were performed to determine whether the salts obtained by the slow evaporation technique can also be prepared using other techniques. The calculated PXRD patterns obtained from LAZYPULVERIX were comAll chemicals were purchased from Sigma Aldrich and were used as received. K. Diffraction data for all compounds were collected on a Bruker APEX II diffractometer . SADABS was usedA D2 PHASER Bruker diffractometer with Cu-K\u03b1 radiation (1.54184 \u00c5) was used for PXRD. The voltage tube and current were at 30 kV and 10 mA max, respectively, with a scintillation counter 1-dim LYNXEYE detector. The scanning process of each sample was between 4\u201350\u00b0 2\u03b8.\u22121.Spectra were obtained from the universal attenuated total reflectance (UTAR) infrared spectrometer Perkin Elmer spectrum two. Sample spectra were measured over the range of 4000\u2013400 cm\u22121. These analyses were conducted from 303\u2013573 K with a heating rate of 10 K min\u22121.DSC analyses were performed on a Perkin-Elmer 6 system with a purge of nitrogen at 20 mL minSamples of 2\u20135 mg were removed from the mother liquor, dried with filter paper, then crushed to a fine powder and placed in a vented pan for the DSC analysis.\u2212)(DBM+). The Cl\u00b7\u00b7\u00b7Cl halogen bond was only found in (Salts of CHPAA with selected linear and aromatic amines were successfully prepared. The formation of the new solid forms was validated using DSC analysis, and in the case of the aromatic amines, the resulting salts demonstrated enhanced thermal stability compared to the starting materials. FTIR spectroscopy confirmed salt formation. The salt structures of CHPAA, except for ("} +{"text": "The dihedral angles between the indole ring system and pendant nitro\u00adbenzodioxolane rings system and phenyl\u00adsulfonyl ring are 4.81\u2005(14) and 72.24\u2005(19)\u00b0, respectively. In the crystal, the indole mol\u00adecules are linked to each other and to the chloro\u00adform mol\u00adecule by weak C\u2014H\u22efO, C\u2014H\u22efCl, C\u2014H\u22ef\u03c0, C\u2014Br\u22ef\u03c0 and C\u2014Cl\u22ef\u03c0 and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions.The title indole derivative crystallizes with a partial occupancy [0.585\u2005(4)] CHCl 24H17Br3N2O6S, crystallizes with a partial occupancy [0.585\u2005(4)] CHCl3 solvent mol\u00adecule. The dihedral angles between the indole ring system and pendant nitro\u00adbenzodioxolane rings system and phenyl\u00adsulfonyl ring are 4.81\u2005(14) and 72.24\u2005(19)\u00b0, respectively. In the crystal, the indole mol\u00adecules are linked to each other and to the chloro\u00adform mol\u00adecule by weak C\u2014H\u22efO, C\u2014H\u22efCl, C\u2014H\u22ef\u03c0, C\u2014Br\u22ef\u03c0 and C\u2014Cl\u22ef\u03c0 and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions. A Hirshfeld surface analysis was carried out and the inter\u00admolecular contacts with the most significant contributions are H\u22efO/O\u22efH (24.3%), H\u22efH (18.4%), Br\u22efH/H\u22efBr (16.8%) and C\u22efH/H\u22efC (8.4%).The title indole derivative, C The C1\u2014N1 and C4\u2014N1 bond lengths are 1.423\u2005(5) and 1.427\u2005(5)\u2005\u00c5, respectively, while in the case of N atoms in planar configurations, the reported mean value is 1.355\u2005(14)\u2005\u00c5 vinyl moiety occurs due to an addition reaction with Br2. The Br1\u2014C16\u2014C15\u2014Br2 grouping is in a trans configuration and the torsion angle has a value of 178.14\u2005(17)\u00b0. Intra\u00admolecular C5\u2014H5\u22efO1, C15\u2014H15\u22efO2, C16\u2014H16\u22efO6 and C24\u2014H24B\u22efBr2 inter\u00adactions \u00b0 with the C1\u2013C8/N1 indole ring system Fig.\u00a01. The C1\u2014ns Fig.\u00a01 are obse3.Cg2i cyclo\u00adhex-3-en-1-yl}vin\u00adyl)-3-methyl-1-(phenyl\u00adsulfon\u00adyl)-1H-indole tetra\u00adhydrate , H\u22efH (18.4%), Br\u22efH/H\u22efBr (16.8%) and C\u22efH/H\u22efC (8.4%).The Hirshfeld surface analysis and the associated two-dimensional fingerprint plots were determined using the 6.E)-3-methyl-2-[2-vin\u00adyl]-1-(phenyl\u00adsulfon\u00adyl)-1H-indole and N-bromo\u00adsuccinimide in dry CCl4 (100\u2005ml) containing a catalytic amount of azobisisobutyro\u00adnitrile was refluxed for 2\u2005h. The reaction mixture was cooled to room temperature. Then, the suspended succinimide was filtered off and the filtrate was concentrated in vacuo to obtain the crude product, which upon trituration with methanol (10\u2005ml) gave the title compound as a bright-yellow solid. Yield: 800\u2005mg (88%) m.p. 431\u2013433\u2005K. The synthesized compound was crystallized by slow evaporation using chloro\u00adform as solvent.A solution of I. DOI: 10.1107/S2056989023007120/hb8071Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023007120/hb8071Isup3.cmlSupporting information file. DOI: 2279852CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Correction: BMC Cancer 23, 540 (2023)https://doi.org/10.1186/s12885-023-10997-xFollowing publication of the original article , the autThe incorrect author name is: \u00c1d\u00e1ny R\u00f3zaThe correct author name is: R\u00f3za \u00c1d\u00e1nyThe author group has been updated above and the original article has been"} +{"text": "Inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds connect individual mol\u00adecules into layers extending parallel to (100). These layers are connected by C\u2014H\u22ef\u03c0 inter\u00adactions. 32H29N5O2\u00b7C3H7NO, the bi\u00adcyclo[3.3.1]nonane ring sys\u00adtem adopts a half-chair/twist-boat conformation, with the phenyl rings in equatorial orientations with respect to the piperidine ring. The two oxane rings of the 2-oxabi\u00adcyclo\u00ad[2.2.2]octane ring system exhibit a distorted boat conformation. Inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds connect the mol\u00adecules in the crystal, generating layers extending parallel to (100). These layers are connected by C\u2014H\u22ef\u03c0 inter\u00adactions. A Hirshfeld surface analysis was per\u00adformed to qu\u00adantify the contributions of the different inter\u00admolecular inter\u00adactions, indicating that the most important contributions to the crystal packing are from H\u22efH (52.5%), N\u22efH/H\u22efN (19.2%), C\u22efH/H\u22efC (18.8%) and O\u22efH/H\u22efO (8.3%) inter\u00adactions.In the title compound, C The phenyl rings are in equatorial orientations with respect to the piperidine ring (C1/C2/N3/C4/C5/C9). The two oxane rings (O9/C9/C1/C8/C7/C10 and O9/C9/C5/C6/C7/C10) of the 2-oxabi\u00adcyclo\u00ad[2.2.2]octane ring system (C10/O9/C9/C1/C8/C7/C6/C5) exhibit a distorted boat conformation with puckering parameters QT = 0.799\u2005(2)\u2005\u00c5, \u03b8 = 91.88\u2005(14)\u00b0, \u03c6 = 247.89\u2005(15)\u00b0 for the O9/C9/C1/C8C7/C10 ring, and QT = 0.826\u2005(2)\u2005\u00c5, \u03b8 = 96.04\u2005(14)\u00b0, \u03c6 = 50.59\u2005(15)\u00b0 for the O9/C9/C5/C6/C7/C10 ring.The mol\u00adecular structure of the title compound is displayed in Fig.\u00a02le Fig.\u00a02. As showle Fig.\u00a02 are QT 3.In the crystal, inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds Table\u00a01 link indCrystalExplorer17 to 1.4361 (blue) a.u. is shown in Fig.\u00a06dnorm surface represent the aforementioned C\u2014H\u22efO and C\u2014H\u22efN inter\u00adactions points from all the contacts contributing to the Hirshfeld surface analysis in normal mode for all atoms. The most important inter\u00admolecular inter\u00adactions are H\u22efH contacts, contributing 52.5% to the overall crystal packing. Other inter\u00adactions and their respective contributions are N\u22efH/H\u22efN (19.2%), C\u22efH/H\u22efC (18.8%), O\u22efH/H\u22efO (8.3%), N\u22efN (0.6%), C\u22efN/N\u22efC (0.3%), C\u22efC (0.2%) and C\u22efO/O\u22efC (0.1%), respectively.Fig.\u00a07et al., 2015The Hirshfeld surface study verifies the significance of H-atom inter\u00adactions in the packing formation. The contributions of H\u22efH and N\u22efH/H\u22efN inter\u00adactions imply that van der Waals inter\u00adactions are important in the crystal packing -3-aza\u00adbicyclo\u00ad[3.3.1]nonan-9-ol (III) and (III) crystallize in monoclinic space groups , whereas (II) is ortho\u00adrhom\u00adbic . In each of the three structures, the bicyclic ring system adopts a chair/chair conformation and the phenyl rings are in equatorial orientations with respect to the piperidine ring. In (II), apart from van der Waals forces, only weak inter\u00admolecular C\u2014H\u22efO-type inter\u00adactions are involved in the packing.Compounds (IV) has monoclinic symmetry and has two mol\u00adecules in the asymmetric unit. A et al., 1995via N\u2014H\u22efO hydrogen bonds.The structure of (V), which likewise is monoclinic , the bicyclic ring system adopts a twin-chair conformation. The two methyl groups attached to the bicycle are in an equatorial orientation for both rings. In the crystal, very long N\u2014H\u22efO hydrogen bonds connect the mol\u00adecules into a chain perpendicular to [010].In solution.The title compound was synthesized using a previously reported procedure (Mamedov 6.Uiso(H) = 1.2 or 1.5Ueq(C). The N-bound H atoms were located from difference-Fourier maps and refined with free atomic coordinates and Uiso = 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989023001718/wm5672sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989023001718/wm5672Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989023001718/wm5672Isup3.cmlSupporting information file. DOI: 2244417CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S is established as a function of several basic geometric parameters to well characterize the noncovalent interaction energy, which exhibits a good inverse correlation with the reorganization energies of the photo-excited states or electron-pumped charged states in organic/polymeric semiconductors. In particular, the experimental 1H, 77Se, and 125Te NMR, the optical absorption and emission spectra, and single crystal structures of eight compounds fully confirm the theoretical predictions. This work provides a simple descriptor to characterize the strength of noncovalent intramolecular interactions, which is significant for molecular design and property prediction.In recent years, intramolecular noncovalent interaction has become an important means to modulate the optoelectronic performances of organic/polymeric semiconductors. However, it lacks a deep understanding and a direct quantitative relationship among the molecular geometric structure, strength of noncovalent interaction, and optoelectronic properties in organic/polymeric semiconductors. Herein, upon systematical theoretical calculations on 56 molecules with and without noncovalent interactions , we reveal the essence of the interactions and the dependence of its strength on the molecular geometry. Importantly, a descriptor Intramolecular noncovalent interactions can block molecules in a given conformation enhancing performance of organic semiconductors. Here, the authors report a molecular descriptor to weigh them that strongly correlates with the reorganization energy of excited or charged states. Many experimental and theoretical works focus on intermolecular noncovalent interactions since they are directly responsible for the formation of molecular crystals, large clusters, folding of proteins, and binding of DNA and drugs14. However, intramolecular noncovalent interactions are less investigated, although they are also important to determine the structure of basic units for building larger aggregates17. It is noteworthy that the intramolecular interactions significantly influence the conformation of the organic/polymeric semiconductors, which is critical to determine their physicochemical properties, such as optical properties and charge transport mobilities20. Many groups have adopted the noncovalent intramolecular interactions as an important strategy for designing high-performance organic/polymeric semiconductors for different applications, including organic solar cells (OSCs)28, thin film transistors (OTFTs)32, photodetectors (OPDs)33 and light-emitting diodes (OLEDs)37, since Huang, Marks, Facchetti, and coworkers termed this intramolecular noncovalent interaction as \u201cNoncovalent Conformational Locks (NoCLs)\u201d owing to their conformation-locking feature in enhancing the planarity and rigidity of organic semiconductors38. For example, the noncovalent fused ring electron acceptors (NFREAs) have been developed using the NoCLs strategy, which greatly improved the power conversion efficiencies (PCEs) to reach over 15%, similar to those of the fused ring electron acceptors (FREAs), while significantly decreased the synthetic complexity28. Moreover, various NoCLs have been used to enhance the charge transport mobilities of organic/polymeric semiconductors for OTFTs29, affording the record mobility as high as 14.9 cm2\u00a0V\u22121\u00a0s\u22121\u200931. Also, the OLED based on the thermally activated delayed fluorescence (TADF) obtained by the NoCLs strategy showed a high external quantum efficiency of 23.2%39. Although these investigations presented the efficacy of NoCLs in designing high-performance organic/polymeric semiconductors, they lack direct correlation between chemical structures, the strengths of NoCLs, and the optoelectronic properties of organic/polymeric semiconductors. Furthermore, the nature of intramolecular NoCLs in organic/polymeric semiconductors, whether they arise from orbital overlap or electrostatic interactions, remains controversial47.Noncovalent interactions play important roles in chemistry, biology, catalysis, and material sciences49, which dramatically increased the difficulty of experimental characterization. Moreover, due to various chemical elements or groups, there is a wide variety of noncovalent intramolecular interaction, such as cation-\u03c0, anion-\u03c0, chalcogen-\u03c0, carbonyl-\u03c0, hydrogen bonding, halogen bonding, and chalcogen-bonding in organic molecules53. Thus, the theoretical study of these interactions became very complicated and challenging.Although many efforts have been made, it is very challenging to deeply investigate the fundamentals of NoCLs. First, noncovalent interactions are very weak forces, usually one to two orders of magnitude smaller than covalent interactions1H, 77Se, and 125Te NMR, the optical absorption and emission spectra, and the single crystal structures of eight synthesized compounds fully confirmed the theoretical predictions.In this work, 56 molecules Fig.\u00a0 with varM(S\u00b7\u00b7\u00b7O) and PhM/PhM-H(S\u00b7\u00b7\u00b7O) were selected as representatives to investigate the feature of X\u00b7\u00b7\u00b7Y NoCLs. The relaxed potential energy (E) surface (PES) scans were performed along the dihedral angle \u03b8 analysis54 was performed to obtain the variation of energy (E(2)), and took atomic dipole moment-corrected Hirshfeld (ADCH) charge55 , and Eelec, as well as the distances between the atoms. From Fig.\u00a0M(S\u00b7\u00b7\u00b7O) appeared at \u03b8\u2009=\u20090.01o, whose E was set to zero on the PES. At equilibrium geometry, the S\u00b7\u00b7\u00b7O distance d(S\u00b7\u00b7\u00b7O) of 2.85\u2009\u00c5 is much shorter than the sum of van der Waals radii (dv) of S and O atoms dv(S\u00b7\u00b7\u00b7O) of 3.32\u2009\u00c5; the Eelec is a small negative value due to few charges on S (qS) and O (qO) atoms is a large negative value owing to large overlap between the n-orbital (lone pair electron of the oxygen atom) and the \u03c3*-orbital of the S-C bond (n(O)\u2009\u2192\u2009\u03c3*(S-C)) are formed by the orbital interactions. Differently, the two minima on the PES of type II are very close in energy with a large energy barrier with 0o\u2009\u2264\u2009\u03b8\u2009\u2264\u200990o, and the H\u00b7\u00b7\u00b7O interaction is detected in PhM-H(S\u00b7\u00b7\u00b7O) with 90o\u2009<\u2009\u03b8\u2009\u2264\u2009180o. As shown in Fig.\u00a0PhM(S\u00b7\u00b7\u00b7O) (\u03b8\u2009=\u200918.92o and d(S\u00b7\u00b7\u00b7O)\u2009=\u20092.76\u2009\u00c5), and both the orbital interactions and the electrostatic interactions are weak at the minimal of PhM-H(S\u00b7\u00b7\u00b7O) (\u03b8\u2009=\u2009157.34o and d(H\u00b7\u00b7\u00b7O)\u2009=\u20092.31\u2009\u00c5). The orbital interaction in PhM(S\u00b7\u00b7\u00b7O) comes from n(O)\u2192\u03c3*(S-C) and that in PhM-H(S\u00b7\u00b7\u00b7O) stems from n(O)\u2192\u03c3*(H-C).The molecules 5 Figure\u00a0 to calcuenergy (E), and toenergy (E), and tos Figure\u00a0; and the) Figure\u00a0. These iier Fig.\u00a0. Hence, 56 for the studied systems. For type II systems, the energy differences between two minima (PhM-H(O\u00b7\u00b7\u00b7Y) is more stable than PhM(O\u00b7\u00b7\u00b7Y), while PhM(X\u00b7\u00b7\u00b7O) (X\u2009=\u2009S and Se) and PhM(Te\u00b7\u00b7\u00b7Y) are more stable than their corresponding PhM-H. The differences in energies between two conformations are small for the others. The potential energy surfaces of the systems were further scanned at the same level and plotted in Figure\u00a0PhM(X\u00b7\u00b7\u00b7C) and PhM-H(X\u00b7\u00b7\u00b7C) , owing to a low energy barrier of mutual transformation between them, and the others have only one conformation due to large energy barrier.The nature of NoCLs was examined at the equilibrium geometry of the studied systems. The equilibrium geometries were optimized and the frequencies were calculated at the B3LYP(D3)/6-31\u2009+\u2009G(d) levelsee Fig.\u00a0. It is oM(X\u00b7\u00b7\u00b7Y) ones, 9 PhM(X\u00b7\u00b7\u00b7Y) ones, and 11 PhM-H(X\u00b7\u00b7\u00b7Y) ones). It is generally believed that the NoCLs are effectively formed between two atoms if the distance between them is less than dv. According to the \u0394d\u2009=\u2009dv-d results given in Fig.\u00a0d (>0.10\u2009\u00c5), being conducive to the formation of NoCL. The nature of NoCLs is either electronic interaction or orbital interaction45. From Fig.\u00a0\u22121, which is impossible to be responsible for the NoCLs. The orbital interaction varies greatly with E(2) ranging from \u22125.27 to 0.00\u2009kcal\u2009mol\u22121 for the systems with and without NoCLs, which is likely to be the main contributor to form NoCLs. Of 36 compounds, 15 with E(2)<\u22121.00\u2009kcal\u2009mol\u22121 are considered to have significant NoCLs, and the strength of NoCL exhibits the order of S\u00b7\u00b7\u00b7F\u2009<\u2009Se\u00b7\u00b7\u00b7F\u2009<\u2009Te\u00b7\u00b7\u00b7F, S\u00b7\u00b7\u00b7O\u2009<\u2009Se\u00b7\u00b7\u00b7O\u2009<\u2009Te\u00b7\u00b7\u00b7O, and X\u00b7\u00b7\u00b7F\u2009<\u2009X\u00b7\u00b7\u00b7O . The orbital interaction is determined by the type, shape, and orientation of orbitals and bonds. As shown in Fig.\u00a02), n(p)\u2192\u03c3*(sp2), and n(sp)\u2192\u03c3*(sp2) for M and PhM molecules, and n(s)\u2192\u03c3*(s), n(p)\u2192\u03c3*(s) and n(sp2)\u2192\u03c3*(s) for PhM-H molecules. The n(sp2)\u2192\u03c3*(sp2) shows the strongest interaction when p-orbital component dominates in the hybrid sp2-orbital, because the shape and orientation of the sp2-orbital are favorable for overlap, such as Te\u00b7\u00b7\u00b7O NoCL. While the participation of s-orbital weakens the interaction due to its very small electronic density distribution area, like H\u00b7\u00b7\u00b7Y NoCL. Hence, the H\u00b7\u00b7\u00b7Y NoCLs in PhM-H molecules are always weaker than X\u00b7\u00b7\u00b7Y NoCLs in M and PhM molecules. Overall, 15 of 36 molecules were predicted to have significant NoCLs, namely, X\u00b7\u00b7\u00b7Y and Te\u00b7\u00b7\u00b7S in M and PhM, and H\u00b7\u00b7\u00b7S and H\u00b7\u00b7\u00b7Cl in PhM-H and the length of NoCL d(S\u00b7\u00b7\u00b7O), the angle between S\u00b7\u00b7\u00b7O NoCL and S-C bond (\u03b1), and the dihedral angle between two thiophene rings (\u03b8) which is closely related to molecular structural planarity and disorder57. It is found the E(2) as an exponential function of the distance, and the cosine function of the angle \u03b1 or \u03b8. Based on this, we proposed a simple descriptor that contains several geometrical parameters versus S, which are calculated based on the optimized equilibrium geometries for 36 isolated systems in Fig.\u00a0R2 is as high as 0.927, which indicates a very strong positive correlation between E(2) and S. Moreover, the S value varies in the order of S\u00b7\u00b7\u00b7O\u2009<\u2009Se\u00b7\u00b7\u00b7O\u2009<\u2009Te\u00b7\u00b7\u00b7O and S\u00b7\u00b7\u00b7F\u2009<\u2009Se\u00b7\u00b7\u00b7F\u2009<\u2009Te\u00b7\u00b7\u00b7F, which is in good agreement with the order of strength of NoCLs obtained above by E(2). This is mainly because from S to Te the atomic radius gradually increases while the distances of X\u00b7\u00b7\u00b7Y and other factors are almost unchanged, which results in the increase of \u0394d, corresponding to the strengthening of NoCLs in this order. In addition, as it is described above that the orbital interactions of hydrogen bond in PhM-H compounds happen between s-orbital of H atom and p- or sp2-orbital of Y atoms, and it is almost independent of the orientation of C-H bond because s-orbital is spherical. And if removing cos\u03b1 from the S for PhM-H systems, the results would become better and S were further calculated for 11 compounds of 36 with strong NoCLs at B3LYP(D3)/def2-TZVP, \u03c9B97XD/def2-TZVP and CCSD(T)/def2-TZVP levels and S becomes stronger at large basis set and higher theory levels. Based on the optimized geometries by B3LYP(D3)/def2-TZVP, the linear fitting coefficient is improved from R2\u2009=\u20090.974 at B3LYP(D3)/def2-TZVP level to R2\u2009=\u20090.991 at CCSD(T)/def2-TZVP level. Based on the optimized geometries by \u03c9B97XD/def2-TZVP and S with R2\u2009=\u20090.992 and S of 56 compounds is still strong with R2\u2009=\u20090.907 less than ca. \u22121.00\u2009kcal\u2009mol\u22121).Considering the effect of the basis set and method on the results, the els Fig.\u00a0c, d. TheP Figure\u00a0, the CCS992 Fig.\u00a0. In addi7 Figure\u00a0. Therefoa) absorbs a phonon to form an excited state (b), the excited molecule firstly experiences a relaxation of geometrical structure to a new equilibrium point (c) by releasing reorganization energy (\u03bbES) and then decays to the ground state (d) by radiative and nonradiative processes, finally relax to the initial ground-state equilibrium point (a) by giving out reorganization energy (\u03bbGS). The reorganization energy decides the shape and width of the optical spectra. Under the framework of the linear coupling model, the sum of the two reorganization energies can be crudely considered as the Stokes shift between the absorption and emission spectra, \u03ba\u2009=\u2009\u03bbGS\u2009+\u2009\u03bbES59. Moreover, there is a positive relationship between the nonradiative decay rate and the excited reorganization energy, in which the excitation energy is always much larger than the reorganization energy61. Consequently, small reorganization energy can effectively block the nonradiative decay process. For electron-pumped charge transfer processes, the charge transfer rates between molecule dimers can be calculated by Marcus rate equation in the high temperature and short time limits, +\u2192M+\u2009+\u2009M), \u0394G can be approximately zero, and there is an inverse relationship between the rate constant of charge transfer and the charge reorganization energy \u03bbcharge. As a result, the molecules with small reorganization energy usually are designed to exhibit excellent charge transport property62. The \u03bbcharge can be calculated by the sum of \u03bbNS and \u03bbCS between the neutral (ground) state (NS) and the charged states (CS) in organic molecules, as shown in Fig.\u00a0As it is well known that reorganization energy or relaxation energy is an important physical parameter to character the optoelectronic properties. Reorganization energy is defined as the energy dissipated from the equilibrium geometry in the initial state to the relaxed equilibrium geometry in the final state for an isolated molecule and PhM(X\u00b7\u00b7\u00b7C) with X\u2009=\u2009O, S, Se, and Te were calculated as shown in Fig.\u00a0S behaves good negative relationship with \u03bbopt (Stokes shift \u03ba). The Stokes shifts of the systems with strong NoCLs are far smaller than those of the corresponding systems without NoCLs. Moreover, the larger S is, the larger the difference of Stokes shift is. For example, the Stokes shift from PhM(Se\u2009\u2219\u2009\u2219\u2219C) to PhM(Se\u2009\u2219\u2009\u2219\u2219O) is decreased by 75\u2009nm, while that from PhM(Te\u2009\u2219\u2009\u2219\u2219C) to PhM(Te\u2009\u2219\u2009\u2219\u2219O) decreased by 118\u2009nm. The linear correlation between \u0394\u03bbopt and S is very good with R2\u2009=\u20090.801 for 6 pairs of compounds of type II molecules with apparent noncovalent interactions of X\u2009=\u2009O, S, Se, and Te, and Y\u2009=\u2009O and C were synthesized ,PhM(S\u00b7\u00b7\u00b7O) PhM(Se\u00b7\u00b7\u00b7O), and PhM(Te\u00b7\u00b7\u00b7O). In these compounds, the d values are 2.38(9) \u00c5 for H\u00b7\u00b7\u00b7O, 2.68(2) \u00c5 for S\u00b7\u00b7\u00b7O, 2.69(1) \u00c5 for Se\u00b7\u00b7\u00b7O, and 2.76(0) \u00c5 for Te\u00b7\u00b7\u00b7O, respectively, which are much shorter than the corresponding dv, confirming the existence of strong NoCLs. Differently, both PhM/PhM-H (S\u00b7\u00b7\u00b7C) and PhM/PhM-H (Se\u00b7\u00b7\u00b7C) possess two conformations, which fully prove the theoretical predictions above. Moreover, the torsion angles between chalcogenide and phenyl rings in PhM(S\u00b7\u00b7\u00b7O) and PhM(Se\u00b7\u00b7\u00b7O) are much smaller than those in PhM(S\u00b7\u00b7\u00b7C) and PhM(Se\u00b7\u00b7\u00b7C) , illustrating the \u201clock of planarity\u201d function of NoCLs.To confirm the theoretical prediction, eight compounds d Figure\u00a0\u2013S18, andd Figure\u00a0. The con1H-1H NOESY NMR of eight compounds was measured in d2-tetrachloroethane solution, which is always used to certify conformational preferences through spatial interactions between different 1H signals is more stable than PhM(O\u00b7\u00b7\u00b7O) in d2-tetrachloroethane and the O\u00b7\u00b7\u00b7H NoCL is stronger than O\u00b7\u00b7\u00b7O one. Similarly, the cross-peak between H1 and H2 in Fig.\u00a0PhM(Te\u00b7\u00b7\u00b7O) conformation with strong Te\u00b7\u00b7\u00b7O NoCLs. Both the cross-peaks between H5 and H2, and between H1 and H2 are observed in Fig.\u00a0PhM(O\u00b7\u00b7\u00b7C) and PhM-H(O\u00b7\u00b7\u00b7C), and PhM(Te\u00b7\u00b7\u00b7C) and PhM-H(Te\u00b7\u00b7\u00b7C) coexist owing to freely rotatable aromatic rings without NoCLs, suggesting no effective NoCLs in these compounds. These NMR observations are consistent with the experimental results that no single conformation was observed for PhM-H(S\u00b7\u00b7\u00b7C) and PhM-H(Se\u00b7\u00b7\u00b7C) in their respective crystals as shown in Figure\u00a0PhM(S\u00b7\u00b7\u00b7O) seems to be inconsistent with its crystal structure since it exhibits no cross-peaks neither between H6 and H2, nor between H1 and H2 calculated in Table\u00a0\u03b8 of isolated PhM(S\u00b7\u00b7\u00b7O) is calculated to be 17.29o, which is larger than that of 4.68o/5.59o in crystal, implying a more twisted conformation in solution. Since the strengths of NoCLs decrease sharply from \u03b8\u2009=\u20090o to 20o may become very weak in solution, causing the alienation of H1 and H2. Thus, these experimental observations reinforce the reliability of theoretical predictions. In addition, The 77Se and 125Te NMR of PhM(Se\u00b7\u00b7\u00b7O), PhM(Se\u00b7\u00b7\u00b7C), PhM(Te\u00b7\u00b7\u00b7O), and PhM(Te\u00b7\u00b7\u00b7C) are further measured and the results were shown in Figure\u00a077Se NMR of PhM(Se\u00b7\u00b7\u00b7O) displays one single peak of symmetric Se nucleus located at 645.70 ppm, with a downfield shift of 9.15 ppm compared to the signal of PhM(Se\u00b7\u00b7\u00b7C) at 636.55 ppm. Since the inductive effect of the O atom generally decays along chemical bonds and disappears after three \u03b4 bonds, this phenomenon can only be triggered by Se\u00b7\u00b7\u00b7O interaction. Note that this trend is in accordance with that in literature reported by Tomita64. Similarly, the downfield shift phenomenon is observed in PhM(Te\u00b7\u00b7\u00b7O) (851.62 ppm) in comparison to that of PhM(Te\u00b7\u00b7\u00b7C) (841.78 ppm) in 125Te NMR, which reveals the existence of Te\u00b7\u00b7\u00b7O interactions. Overall, the reflected results by 1H-1H NOESY NMR tell us that the Te\u00b7\u00b7\u00b7O, O\u00b7\u00b7\u00b7H, and S\u00b7\u00b7\u00b7O are significant NoCLs while the O\u00b7\u00b7\u00b7C, Te\u00b7\u00b7\u00b7C, and H\u00b7\u00b7\u00b7C are not effective NoCLs in these compounds, which are consistent with the theoretically predicted results.The als Fig.\u00a063. As sh2 Figure\u00a0. AccordiPhM(Te\u00b7\u00b7\u00b7O) with strong NoCLs as an example, the 1H-1H NOESY NMR is further measured in d2-dichloromethane (d2-DCM) and d6-dimethylsulfoxide (d6-DMSO) shown in Figure\u00a01 and H2 appears in the three solvents. The position of the cross-peak hardly shifts with the increase of the polarity of the solvent, which implies these NoCLs are not controlled by the electrostatic interactions47.In order to reveal the nature of NoCLs, the effect of solvents on NoCLs was investigated. Taking PhM(X\u00b7\u00b7\u00b7C) , there display immense redshifts of the absorption peaks in PhM(X\u00b7\u00b7\u00b7O) , which accounts for the introduction of NoCLs for higher molecular planarity65. The resultant Stokes shift sharply decreased from PhM(X\u00b7\u00b7\u00b7O) to PhM(X\u00b7\u00b7\u00b7C) , which depicts the enhancement of rigidity brought by NoCLs. Specific \u0394\u03ba values are rendered to estimate the relative ability of NoCLs in altering rigidity with 61 (63) nm for S\u00b7\u00b7\u00b7O interaction, 75 (66) nm for Se\u00b7\u00b7\u00b7O interaction, and 118 (91) nm for Te\u00b7\u00b7\u00b7O interaction. This implies the stronger the NoCLs, the more its ability in enhancing rigidity. Moreover, the intensity order of S\u00b7\u00b7\u00b7O\u2009<\u2009Se\u00b7\u00b7\u00b7O\u2009<\u2009Te\u00b7\u00b7\u00b7O of the NoCLs is perfectly reflected by the \u0394\u03ba values, which are in good agreement with the theoretically calculated results above. The same experiments are carried out in tolune and THF solvents with different polarity, and the Stokes shifts and their changes exhibit extremely small changes based on the theoretical equilibrium structures in the gas (solid) phase and the experimental S (SExp) based on the experimental crystal structures for the six compounds , which demonstrates that the SExp based on the crystal structure is also very reliable. To confirm the assumption, we arbitrarily select a series of compounds from the CCDC crystal database to calculate E(2) and the SExp based on experimental crystal structures and plot them in Fig.\u00a0R2 is very high with a value of 0.972, which reveals that the S can be applied widely. Thus far, it is safe to conclude that S is a suitable descriptor for evaluating the strength of NoCLs, which would be used for further machine-learning-based molecular screening studies.To extend the application of this descriptor, twelve compounds were selected to be investigated Fig.\u00a0. First, nds Fig.\u00a0. Excitin1H, 77Se, and 125Te NMR, and UV-vis spectra of the eight new synthesized systems fully confirmed the theoretical predictions. This work provides an in-depth understanding and simple description of noncovalent interaction, which is important for the rational molecule design of high-performance organic/polymeric semiconductors.Herein, 56 organic semiconductors with and without noncovalent interactions were investigated to show that the interactions are mainly derived from orbital interactions based on the theoretical calculations and experimental results. The strength of interactions followed the order of S\u00b7\u00b7\u00b7F\u2009<\u2009Se\u00b7\u00b7\u00b7F\u2009<\u2009Te\u00b7\u00b7\u00b7F, S\u00b7\u00b7\u00b7O\u2009<\u2009Se\u00b7\u00b7\u00b7O\u2009<\u2009Te\u00b7\u00b7\u00b7O, and X\u00b7\u00b7\u00b7F\u2009<\u2009X\u00b7\u00b7\u00b7O . In contrast, O\u00b7\u00b7\u00b7Y , X\u00b7\u00b7\u00b7C , and Se\u00b7\u00b7\u00b7Y cannot form effective noncovalent interactions in the systems. Furthermore, it is disclosed that the strength of noncovalent interactions is closely related to the structural parameters, including the molecular planarity, the angle between two relevant orbitals, and the distance between the two related atoms. Significantly, a descriptor 66 of theory using the basis set 6-31\u2009+\u2009G(d) and LANL2DZ for Se and Te or def2-TZVP in Guassian16 Program67, including geometrical parameters, energy, atomic dipole moment-corrected Hirshfeld population (ADCH) charge analyses68, natural bond orbital (NBO) analysis54. NBO analysis have been carried out using Multiwfn software69. The CCSD/def2-TZVP method was used to calculate the orbital interaction. As for the solid phase, the initial structure is obtained from the X-ray crystal structure, and geometrical structures in the ground state are calculated based on QM/MM model through Gaussian16 Program. The centered molecule is treated as a high layer and calculated at B3LYP(D3)/def2-TZVP level for QM, and the surrounding molecules are treated as a low layer and computed by MM with UFF forces field. The reorganization energy is calculated using the adiabatic potential energy surface method in MOMAP72.All the geometrical and electronic structures of the investigated system in the ground state were calculated at B3LYP(D3) or \u03c9B97XD level3)4 (5% equiv) were added under nitrogen in anhydrous toluene (20.0\u2009mL). The mixture was stirred and refluxed at 120\u2009\u00b0C for 12\u2009h. Afterwards, an aqueous potassium fluoride solution was added to the mixture. After quenching the reaction, the organic phase was extracted with dichloromethane (3\u2009\u00d7\u2009100\u2009mL). The combined organic portion was collected, washed with water and brine, and dried over MgSO4. After filtration, the solution was filtered through a short silica gel column (petroleum ether), concentrated to afford the crude product, which is subjected to the recrystallization in hexane. Finally, the crystals were dried in vacuo to afford PhM(X\u00b7\u00b7\u00b7O)/PhM(X\u00b7\u00b7\u00b7C). The detailed synthesis is described in Supplementary Information, and the final products were characterized by 1H-NMR, 13C-NMR, 77Se-NMR, and 125Te-NMR. (Supplementary Section 13).Preparations of 1,4-dihexyl-2,5-diiodobenzene, 1,4-dihexyloxy-2,5-diiodobenzene, 2-tributylstannyl-derivatives are illustrated thoroughly in Supplementary Information. To a round-bottom flask (100\u2009mL), 1,4-dihexyl-2,5-diiodobenzene (1.0 equiv), 2-tributylstannyl-derivatives (2.5 equiv) and Pd(PPhUV-Vis absorption spectra were measured on a Gary 60 UV-Vis Spectrophotometer. Photoluminescence spectra were measured on a FLS 1000 (EDINBURGH INSTRUMENTS) with an Xenon Lamp. All liquid samples were well dissolved in chloroform. All film samples were spin-coated on glass substrates.A glass vial (5\u2009mL) containing a chloroform or toluene solution of the compounds (2\u2009mg) was placed inside a vial (20\u2009mL) containing methanol. After 2\u20137 days, white (yellow) solid single crystals were collected.Supplementary InformationPeer Review FileDescription of Additional Supplementary FilesSupplementary Data 1Supplementary Data 2Supplementary Data 3Supplementary Data 4Supplementary Data 5Supplementary Data 6Supplementary Data 7"} +{"text": "The PdII atom displays a slightly distorted square-planar PdP2S2 geometry with a bidentately coordinated pyridin-2-ylcarbonimidodithio\u00adate ligand and two triphenyl\u00adphosphine mol\u00adecules, coordinated in cis positions. The crystal structure features weak \u03c0\u2013\u03c0 [centroid\u2013centroid distance =3.7327(15)\u2005\u00c5] and C\u2013H\u22ef\u03c0 inter\u00adactions and contains an almost spherically shaped void of 50.4\u2005\u00c53 per unit cell.The title compound, [Pd(C DOI: 10.1107/S1600536810050518/im2253Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Adjacent Zn atoms are bridged by the bidentate carboxyl\u00adate groups into a cationic chain extending along [010]. N\u2014H\u22efN, O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds link the cationic chains, nitrate anions and uncoordinated water mol\u00adecules into a supra\u00admolecular network. \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings and between the pyridine and benzene rings [centroid\u2013centroid distances = 3.615\u2005(4) and 3.636\u2005(4)\u2005\u00c5] are present.In the title compound, {[Zn(C DOI: 10.1107/S1600536811021088/hg5047Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Ru0 atom adopts a distorted trigonal\u2013bipyramidal coordination geometry, with the C\u00a0O ligand and the ketone O atom occupying the axial positions. The two triphenyl\u00adphosphane ligands are cis to each other. The olefinic C=C bond is almost coplanar with the Ru0 atom and the two P atoms (maximum deviation of 0.0516\u2005\u00c5 from the mean plane defined by the five constituent atoms). The coordinated C=C bond has a length of 1.449\u2005(3)\u2005\u00c5, which is significantly longer than that of a free C=C bond (1.34\u2005\u00c5). There are two C\u2014H\u22ef\u03c0 inter\u00adactions involving neighbouring phenyl rings in the mol\u00adecule. In the crystal, mol\u00adecules are linked via two further C\u2014H\u22ef\u03c0 inter\u00adactions.The 1-vinyl\u00adpyrrolidin-2-one ligand in the title compound, [Ru(C For C=C al. 1989. For str al. 1998; Jazzar al. 2001.6H9NO)(C18H15P)2(CO)] = 0.025wR(F2) = 0.084S = 1.157113 reflections442 parametersH-atom parameters constrainedmax = 0.66 e \u00c5\u22123\u0394\u03c1min = \u22120.80 e \u00c5\u22123\u0394\u03c1CrystalClear used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812014766/su2399Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The ZnII ions are bridged by the ligands, forming a helical chain along [001]. C\u2014H\u22efN and C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the imidazole rings [centroid\u2013centroid distance = 3.4244\u2005(10)\u2005\u00c5] assemble the chains into a three-dimensional supra\u00admolecular network.In the title complex, [Zn(C DOI: 10.1107/S1600536812012706/hy2526Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "One of the CuII ions has a square-pyramidal arrangement, which is defined by a water mol\u00adecule occupying the apical position, with the equatorial ligators consisting of two N atoms from a 2,2\u2032-bipyridine mol\u00adecule, one carboxyl\u00adate O atom from a terephthalate ligand and one O atom from a water mol\u00adecule. The other CuII ion has a similar coordination environment, except that the apical position is occupied by a chloride ligand instead of a water mol\u00adecule. An O\u2014H\u22efO and O\u2014H\u22efCl hydrogen-bonded three-dimensional network is formed between the components.In the binuclear title compound, [Cu Cl(C10H8N2)2(H2O)3]NO3\u00b7H2O = 0.052wR(F2) = 0.172S = 1.046973 reflections412 parametersH-atom parameters constrainedmax = 0.71 e \u00c5\u22123\u0394\u03c1min = \u22120.64 e \u00c5\u22123\u0394\u03c1RAPID-AUTO I, global. DOI: 10.1107/S1600536812019848/fj2542Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Intra- and inter\u00admolecular \u03c0\u2013\u03c0 stacking is present in the crystal structure, and the centroid\u2013centroid distances between the benzene and pyridine rings of adjacent dmphen ligands are 3.492\u2005(3) and 3.592\u2005(3)\u2005\u00c5, respectively. Inter\u00admolecular C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions help to stabilize the crystal structure.In the title complex, [Pb(NO DOI: 10.1107/S1600536810022312/xu2779Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Two CdII atoms are bridged by two chloride anions, forming an inversion-related Cd2Cl2 unit; these units are further linked through thio\u00adcyanate anions, leading to a chain structure extending parallel to [010]. Weak \u03c0\u2013\u03c0 stacking inter\u00adactions with centroid\u2013centroid distances of 3.430\u2005(4)\u2005\u00c5 and an inter\u00adplanar separation of 3.390\u2005(3)\u2005\u00c5 between the pyridine and benzene rings link the chains into a two-dimensional network parallel to (C DOI: 10.1107/S1600536811051373/wm2567Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The silver ion is surrounded by two tryptanthrin ligands, each coordinating through the N atoms, with Ag\u2014N bond lengths of 2.247\u2005(3) and 2.264\u2005(3)\u2005\u00c5, and an anionic nitrate ligand coordinating through two O atoms, with Ag\u2014O bond lengths of 2.499\u2005(3) and 2.591\u2005(3)\u2005\u00c5. The N\u2014Ag\u2014N plane and the O\u2014Ag\u2014O plane are roughly perpendicular, making a dihedral angle of 81.6\u2005(2)\u00b0. In the crystal, C\u2014H\u22efO inter\u00adactions between aromatic H atoms and keto and nitrate O atoms as well as \u03c0\u2013\u03c0 inter\u00adactions [centroid-centroid distance = 3.706\u2005(4)\u2005\u00c5] give rise to a three-dimensional network.In the crystal structure of the title compound, [Ag(NO DOI: 10.1107/S1600536812001821/rn2096Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The benzene ring of the 4-carb\u00adoxy-2-sulfonato\u00adbenzoate anion is twisted with respect to the two phen ring systems at dihedral angles of 66.38\u2005(9) and 53.56\u2005(9)\u00b0. In the crystal, inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonding links the mol\u00adecules into chains running parallel to [100]. Inter\u00admolecular \u03c0\u2013\u03c0 stacking is also observed between parallel phen ring systems, the face-to-face distance being 3.432\u2005(6)\u2005\u00c5.In the title complex, [Mn(C DOI: 10.1107/S1600536810023743/xu2781Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 23.00\u2005(10)\u00b0, while the pyridine ring and the benzene ring are oriented at a dihedral angle of 85.34\u2005(4)\u00b0. An intra\u00admolecular O\u2014H\u22efO hydrogen bond occurs between coordinating water mol\u00adecule and the carboxyl\u00adate group. In the crystal, N\u2014H\u22efO, O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular network. A weak C\u2014H\u22ef\u03c0 inter\u00adaction also occurs in the crystal.In the title complex, [Cu(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.025wR(F2) = 0.066S = 1.093482 reflections216 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.40 e \u00c5\u22123\u0394\u03c1min = \u22120.39 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812028814/xu5579Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The complex shows several inter\u00adesting features: (i) the polyoxygenated loop of C4 effectively chelates a sodium cation in the form of a distorted octahedron and separates it from the iodide counter-ion, the shortest Na+\u22efI\u2212 distance being greater than 6.5\u2005\u00c5; (ii) the cavity of C4 is filled by a methanol mol\u00adecule; (iii) a second methanol mol\u00adecule is hydrogen-bonded to the N atom of a pyridinyl substituent pendant of C4 and halogen-bonded to the I atom of a tFdIB mol\u00adecule; (iv) the two I atoms of another tFdIB mol\u00adecule are halogen-bonded to two iodide anions, which act as monodentate halogen-bond acceptorss; (v) one of the two tFdIB molecules is located about a centre of inversion.The title compound, [Na(C When ca62H76N2O6)]I\u00b71.5C6F4I2\u00b72CH4O = 0.045wR(F2) = 0.095S = 1.1333353 reflections847 parametersH-atom parameters constrainedmax = 2.33 e \u00c5\u22123\u0394\u03c1min = \u22121.52 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SIR2002 global, I. DOI: Click here for additional data file.10.1107/S1600536813007757/bg2501Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The crystal packing is stabilized by inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.4669\u2005(19) and 3.764\u2005(2)\u2005\u00c5].In the title compound, (C DOI: 10.1107/S1600536810054371/bt5430Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Partial hydrolysis led to the title compound, the hydro\u00adchloric acid adduct of the dichloride, having a penta\u00adcoordinated Sn atom with a trigonal\u2013bipyramidal C2SnCl3 core. The N atom of the 2-[(diisopropyl\u00adammonio)\u00admeth\u00adyl]phenyl ligand forms a strong intra\u00admolecular N\u2014H\u22efCl hydrogen bond, resulting in a zwitterionic species, [2-(iPr2HN+CH2)C6H4]SnBuCl3 \u2212\u00b7CH2Cl2. Disorder was found in the n-butyl group, which was refined as disordered over three positions, with site occupancies of 0.22\u2005(1), 0.51\u2005(1) and 0.27\u2005(2).The title compound, [Sn(C DOI: 10.1107/S1600536810050713/zl2332Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atom in the polymeric title compound, {[Pb(C13H11N4O)(CH3OH)2]ClO4}n, is coordinated by an N\u2032-[1-(pyridin-2-yl-\u03baN)ethyl\u00adidene]isonicotinohydrazidate ligand via O,N,N\u2032-donors and simultaneously bridged by a neighbouring ligand via the isonicotinoyl N atom; two additional sites are occupied by methanol O atoms. The resultant supra\u00admolecular chain is a zigzag along the c axis. The PbII atom is seven-coordinated within an N3O3 donor set and a lone pair of electrons, which defines a \u03a8-pentagonal\u2013bipyramidal coordination geometry with the pyridine N and lone pair in axial positions. The supra\u00admolecular chains are linked into the two-dimensional array via inter\u00admolecular Pb\u22efN [3.020\u2005(4)\u2005\u00c5] inter\u00adactions. Layers stack along the a axis, being connected by O\u2014H\u22efO hydrogen bonds formed between the coordinated methanol mol\u00adecules and perchlorate anions.The Pb N\u2032-[1-(2-pyrid\u00adyl)ethyl\u00adidene]isonicotinohydrazide ligand, see: Maurya et al. (CH4O)2]ClO4 = 0.024 wR(F 2) = 0.066 S = 1.05 3492 reflections255 parametersH-atom parameters constrainedmax = 0.97 e \u00c5\u22123 \u0394\u03c1min = \u22120.98 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811046769/hg5133Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The equatorial plane is formed by two trans-related N,O-bidentate pyridazine-3-carboxyl\u00adate ligands and the axial positions are occupied by two water mol\u00adecules. The CoII complex mol\u00adecules are stacked in a column along the a-axis direction by an O\u2014H\u22efN hydrogen bond between the non-coordinating pyridazine N atom and the coordinating water mol\u00adecule. These columns are further connected into a layer parallel to the ac plane by additional hydrogen bonds involving the coordinating and non-coordinating water mol\u00adecules, and the non-coordinating carboxyl\u00adate O atom. The crystal packing is completed by inter\u00adlayer weak C\u2014H\u22efO inter\u00adactions.The title compound, [Co(C For a r al. 2004.5H3N2O2)2(H2O)2]\u00b72H2O = 0.026wR(F2) = 0.058S = 1.081369 reflections122 parameters4 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.28 e \u00c5\u22123\u0394\u03c1min = \u22120.31 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: OLEX2 I, global. DOI: 10.1107/S1600536813017340/is5284Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "One of the two quniolinium ions forms an N\u2014H\u22efOp hydrogen bond and the other an N\u2014H\u22efOw (w = water) hydrogen bond. The water mol\u00adecules further link the components by O\u2014H\u22efOp and O\u2014H\u22efOw hydrogen bonds. A number of C\u2014H\u22efO inter\u00adactions and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions [shortest centroid\u2013centroid separation = 3.541\u2005(7)\u2005\u00c5] are also observed. Together, these generate a three-dimensional network.In the title compound, (C DOI: 10.1107/S1600536813027347/hb7146Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atom in the title compound, [Cd(C6H12NS2)2(C10H8N2)], is hexa\u00adcoordinated by two dithio\u00adcarbamate ligands and the N atoms from a bidentate 2,2\u2032-bipyridyl mol\u00adecule. The coordination geometry is based on a distorted trigonal\u2013prismatic arrangement of the N2S4 donor set. Supra\u00admolecular chains, aligned along the a-axis direction, are mediated by C\u2014H\u22efS inter\u00adactions and these are connected into layers that stack along the c axis via \u03c0\u2013\u03c0 inter\u00adactions [Cg(pyrid\u00adyl)\u22efCg(pyrid\u00adyl) = 3.6587\u2005(13)\u2005\u00c5].The Cd DOI: 10.1107/S1600536811006878/pk2304Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions between the phen and DHB ligands [centroid\u2013centroid distances = 3.542\u2005(4) and 3.879\u2005(4)\u2005\u00c5] and between the pyridine and benzene rings of adjacent phen ligands [centroid\u2013centroid distance = 3.751\u2005(4)\u2005\u00c5] stabilize the crystal structure. Intra\u00admolecular O\u2013H\u22efO hydrogen bonds are observed in the DHB ligands.In the mononuclear title complex, [Dy(C DOI: 10.1107/S1600536810048348/hy2381Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There are two acridine mol\u00adecules and one and a half water mol\u00adecules in the asymmetric unit. The half-mol\u00adecule of water is located on a crystallographic twofold axis. The crystal structure is built up from two threads of mol\u00adecule II sewn together with water mol\u00adecules through O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds from one side and with \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.640\u2005(3) and 3.7431\u2005(3)\u2005\u00c5] between overlapping mol\u00adecules II on the other side. Mol\u00adecule I is attached to this thread from both sides by C\u2014H\u22efO hydrogen bonds. The threads are connected to each other by \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.582\u2005(3) and 3.582\u2005(3)\u2005\u00c5] between the inner side of mol\u00adecule I and stabilized by a C\u2014H\u22ef\u03c0 inter\u00adaction on the other side of mol\u00adecule I. This thread with rows of mol\u00adecule I hanging on its sides is generated by translation perpendicular to the a axis.The title compound, C \u00c5 b = 8.893 (5) \u00c5 c = 17.492 (5) \u00c5 V = 4107 (3) \u00c53 Z = 16 K\u03b1 radiationMo \u22121 \u03bc = 0.08 mmT = 197 K 0.3 \u00d7 0.3 \u00d7 0.3 mm Bruker SMART 6000 diffractometer14504 measured reflections3606 independent reflectionsI > 2\u03c3(I)1733 reflections with R int = 0.068 R[F 2 > 2\u03c3(F 2)] = 0.058 wR(F 2) = 0.197 S = 1.00 3606 reflections272 parameters2 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.35 e \u00c5\u22123 \u0394\u03c1min = \u22120.29 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXTL I, global. DOI: 10.1107/S1600536811038220/ez2255Isup2.hkl Structure factors: contains datablock(s) I. DOI: 10.1107/S1600536811038220/ez2255Isup3.cml Supplementary material file. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two imidazole rings are oriented to each other with a dihedral angle of 75.1\u2005(2)\u00b0. Strong O\u2014H\u22efO hydrogen bonds between protonated and deprotonated carboxyl\u00adate groups occur in the mol\u00adecular structure. In the crystal structure extensive O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds help to stabilize the three-dimensional supra\u00admolecular framework. The propyl groups of anions are disordered over two sites with refined occupancies of 0.768\u2005(6):0.232\u2005(6) and 0.642\u2005(8):0.358\u2005(8).In the title complex, [Cd(C DOI: 10.1107/S1600536810031466/xu5001Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One of the CuII ions is coordinated in a distorted square-planar environment and the other is coordinated in a distorted square-pyramidal environment. The long apical Cu\u2014O bond of the square-pyramidal coordinated CuII ion is formed by a symmetry-related O atom, creating a one-dimensional polymer along [010]. In addition, short inter\u00admolecular Cl\u22efCl distances [3.444\u2005(2)\u2005\u00c5] and weak \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.736\u2005(2)\u20133.875\u2005(3)\u2005\u00c5] are observed. The crystal studied was an inversion twin with a refined twin component ratio of 0.60\u2005(1):0.40\u2005(1).The asymmetric unit of the title compound, [Cu DOI: 10.1107/S1600536812028462/lh5494Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II complex, [Cu(C7H4BrO2)2(C6H6N2O)2(H2O)2], contains one half-mol\u00adecule, the CuII atom being located on an inversion center. The unit cell contains two nicotinamide (NA), two 4-bromo\u00adbenzoate (PBB) ligands and two coordinated water mol\u00adecules. The four O atoms in the equatorial plane around the CuII ion form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 22.17\u2005(16)\u00b0, while the pyridine ring and the benzene ring are oriented at a dihedral angle of 82.80\u2005(6)\u00b0. In the crystal, N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network. A weak C\u2014H\u22ef\u03c0 inter\u00adaction is also observed.The asymmetric unit of the title mononuclear Cu N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.039 wR(F 2) = 0.108 S = 1.13 3515 reflections203 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.86 e \u00c5\u22123 \u0394\u03c1min = \u22121.29 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811021696/su2279Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The complex lies on an inversion center. The metal atom is surrounded by two chelating isobidentate O-propan-2-yl (4-eth\u00adoxy\u00adphen\u00adyl)dithio\u00adphospho\u00adnate ligands in a trans configuration binding through the S-donor atoms. The Ni\u2014S bond lengths are 2.2328\u2005(5) and 2.2369\u2005(5)\u2005\u00c5, an insignificant difference to be considered anisobidentate. The Ni\u22efP separation is 2.8224\u2005(5)\u2005\u00c5 and the S\u2014P bond lengths are 2.0035\u2005(7) and 2.0053\u2005(7)\u2005\u00c5. The S\u2014Ni\u2014S (chelating) and S\u2014Ni\u2014S (trans) bond angles are 88.321\u2005(18) and 180\u00b0. The Ni\u2014S\u2014P bond angles are 83.26\u2005(2) and 83.33\u2005(2)\u00b0, indicating a very minor distortion from ideal square-planar geometry for the Ni atom. The P atom, however, is distorted quite significantly from an ideal tetra\u00adhedral geometry, as reflected by the S\u2014P\u2014S and O\u2014P\u2014C bond angles of 101.93\u2005(3) and 100.70\u2005(7)\u00b0, respectively. The title compound, [Ni(C DOI: Click here for additional data file.10.1107/S1600536812045114/bh2460Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The ErIII ion has a distorted dodeca\u00adhedral coordination with six coordinating O atoms derived from the ferrocene\u00adcarboxyl\u00adate ligands and two coordinated O atoms from one water mol\u00adecule and one methanol mol\u00adecule. The asymmetric unit comprises a half of the complex mol\u00adecule and a methanol solvent mol\u00adecule. Intra\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions occur. In the crystal, mol\u00adecules are linked by inter\u00admolecular O\u2014H\u22efO hydrogen bonds and C\u2014H\u22efO as well as C\u2014H\u22ef\u03c0 inter\u00adactions.In the centrosymmetric title coordination compound, [Er DOI: 10.1107/S1600536811051245/kp2371Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Both arsine ligands are equatorial with respect to the Ru3 triangle. In addition, each Ru atom carries one equatorial and two axial terminal carbonyl ligands. The phenyl rings of biphenyl are twisted from each other by dihedral angles of 50.5\u2005(2), 44.5\u2005(2) and 27.8\u2005(2)\u00b0. The arsine-substituted phenyl rings make dihedral angles of 61.56\u2005(18), 89.36\u2005(18) and 83.27\u2005(18)\u00b0 with each other. The dihedral angles between the two benzene rings are 87.5\u2005(2) and 81.95\u2005(19)\u00b0 for the two diphenyl\u00adarsanyl groups. In the crystal, mol\u00adecules are linked into dimers by inter\u00admolecular C\u2014H\u22efO hydrogen bonds. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 [centroid\u2013centroid distance = 3.601\u2005(3)\u2005\u00c5] inter\u00adactions stabilize the crystal structure.In the title DOI: 10.1107/S1600536811000237/ng5094Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds link the mol\u00adecules into columns in [001]. The porous crystal packing is further stabilized via \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings of neighbouring mol\u00adecules [centroid\u2013centroid distance = 3.746\u2005(3)\u2005\u00c5] with voids of 270\u2005\u00c53.In the title compound, [Ni(C DOI: 10.1107/S1600536812014109/cv5274Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The distorted tetrahedral coordination sphere is defined by chelating N atoms that define an acute angle [69.6\u2005(3)\u00b0] and two I atoms that form a wide angle [142.80\u2005(4)\u00b0]. The linearly coordinated HgII atom [177.0\u2005(4)\u00b0] exists with a donor set defined by C and Cl atoms. Secondary inter\u00adactions are apparent in the crystal packing with the tetra\u00adhedrally and linearly coordinated HgII atoms expanding their coordination environments by forming weak Hg\u22efI [3.772\u2005(7)\u2005\u00c5] and Hg\u22efO [2.921\u2005(12)\u2005\u00c5] inter\u00adactions, respectively. Mercury-containing mol\u00adecules stack along the a axis, are connected by \u03c0\u2013\u03c0 inter\u00adactions [inter-centroid distance between pyridine and benzene rings = 3.772\u2005(7)\u2005\u00c5] and define channels in which the dimethyl sulfoxide mol\u00adecules reside. The latter are connected by the aforementioned Hg\u22efO inter\u00adactions as well as C\u2014H\u22efI and C\u2014H\u22efO inter\u00adactions, resulting in a three-dimensional architecture.The title dimethyl sulfoxide solvate, [Hg E)-N-(pyridin-2-yl\u00admethyl\u00adidene)aryl\u00adamine-type ligands, see: Basu Baul, Kundu, H\u00f6pfl et al. I2]\u00b7C2H6OS = 0.053wR(F2) = 0.143S = 1.034848 reflections208 parametersH-atom parameters constrainedmax = 4.40 e \u00c5\u22123\u0394\u03c1min = \u22121.45 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 general, I. DOI: 10.1107/S1600536813029693/hg5358Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The SnIV ion is coordinated by two N atoms [Sn\u2014N = 2.266\u2005(2) and 2.274\u2005(2)\u2005\u00c5] from the bis\u00ad(2-pyrid\u00adyl)selenide ligand and four chloride anions [Sn\u2014Cl = 2.3717\u2005(6)\u20132.3939\u2005(6)\u2005\u00c5] in a distorted octa\u00adhedral geometry. The central six-membered chelate ring has a boat conformation with the Se and Sn atoms deviating by 0.692\u2005(3) and 0.855\u2005(3)\u2005\u00c5, respectively, from the mean plane through the remaining four ring atoms. The pyridine rings are inclined to each other by a dihedral angle of 49.62\u2005(8)\u00b0. The crystal packing exhibits short inter\u00admolecular Se\u22efCl contacts [3.5417\u2005(7) and 3.5648\u2005(7)\u2005\u00c5], weak C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings with a centroid\u2013centroid distance of 3.683\u2005(3)\u2005\u00c5.The title compound, [SnCl DOI: 10.1107/S160053681202586X/cv5309Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal structure, the complex mol\u00adecules and the uncoodinated water mol\u00adecules are connected by O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds into a three-dimensional network. A \u03c0\u2013\u03c0 stacking inter\u00adaction between the pyridyl ring of the ip ligand and the benzene ring of the neighboring ligand [centroid\u2013centroid distance = 3.579\u2005(2)\u2005\u00c5] is also observed.In the title centrosymmetric dinuclear compound, [Mn DOI: 10.1107/S1600536810031909/hy2338Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There are significant differences in the chemically equivalent Cd\u2014O bond lengths [2.618\u2005(2)\u2005\u00c5 and 2.561\u2005(2)\u2005\u00c5].In the title compound, [Cd(C DOI: 10.1107/S1600536811038712/lh5332Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The four O atoms in the equatorial plane around the ZnII cation form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the DENA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 2.96\u2005(11)\u00b0, while the pyridine ring and the benzene ring are oriented at a dihedral angle of 79.26\u2005(4)\u00b0. The coordinating water mol\u00adecule links with the carboxyl\u00adate group via an intra\u00admolecular O\u2014H\u22efO hydrogen bond. In the crystal, O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular network. A \u03c0\u2013\u03c0 contact between the parallel pyridine rings of adjacent mol\u00adecules may further stabilize the crystal structure [centroid\u2013centroid distance = 3.5654\u2005(8)\u2005\u00c5].In the title complex, [Zn(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C10H14N2O)2(H2O)2] = 0.025wR(F2) = 0.081S = 1.164409 reflections246 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.46 e \u00c5\u22123\u0394\u03c1min = \u22120.33 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812031200/xu5586Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the 1-benzyl-3-methyl\u00adimidazolium units, the dihedral angles between imidazolium and phenyl rings are 80.47\u2005(15) and 76.53\u2005(14)\u00b0. The F atoms of the general-position hexa\u00adfluoro\u00adphosphate anion are disordered over two sets of sites in a 0.767\u2005(17):0.233\u2005(17) ratio. In the crystal, the hexa\u00adfluoro\u00adphosphate anions link the cations into three-dimensional networks via inter\u00admolecular C\u2014H\u22efF hydrogen bonds and are further consolidated by \u03c0\u2013\u03c0 stacking [centroid\u2013centroid distances = 3.5518\u2005(15)\u2005\u00c5] inter\u00adactions.In the title compound, K[Ag(C DOI: 10.1107/S1600536810051925/hb5767Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two independent mol\u00adecules do not differ substanti\u00adally and a mol\u00adecule of water completes the asymmetric unit. The synthesis of the title compound does not modify the stereochemical center, as shown by the absolute configuration found in this crystal structure. Comparison with the non-bromo starting material differs mainly by rotation features. For instance the H(methine)\u2014C\u2014C(methyl\u00adene)\u2014C(ipso) is 173.0\u2005(2)\u00b0 torsion angle in one mol\u00adecule and 177.3\u2005(2)\u00b0 in the other, indicating a trans arrangement. This is in contrast with approximately 50\u00b0 in the starting material. A short inter\u00admolecular Br\u22efBr separation is observed [3.2938\u2005(3)\u2005\u00c5]. The molecules in the crystal are connected via a network of hydrogen bonds through an N\u2014H\u22efO hydrogen bond between the hydroxy group of the phenol of the tyrosine and the N\u2014H of the amide of the other molecule and an O\u2014H\u22efO hydrogen bond between the hydroxy group of the carboxylic acid and the oxygen of the carbonyl of the amide.The title compound, C N-Acetyl-3,5-dibromo-l-tyrosine is a substrate of biological inter\u00adest, for instance it is involved in the synthesis of isodityrosine unit, which has been found in numerous biologically active natural products that include K-13, OF4949 and vancomycin family of glycopeptide anti\u00adbiotics. For the synthesis and specific optical activity of the title compound, see: Bovonsombat et al. \u00c5b = 22.5186 (9) \u00c5c = 8.6486 (4) \u00c5\u03b2 = 105.946 (1)\u00b0V = 1331.3 (1) \u00c53Z = 4K\u03b1 radiationMo \u22121\u03bc = 6.10 mmT = 125 K0.23 \u00d7 0.17 \u00d7 0.05 mmBruker SMART CCD area-detector diffractometerSADABS; Bruker 2007Tmin = 0.334, Tmax = 0.750Absorption correction: multi-scan (18331 measured reflections7067 independent reflectionsI > 2\u03c3(I)6449 reflections with Rint = 0.030R[F2 > 2\u03c3(F2)] = 0.022wR(F2) = 0.048S = 0.927067 reflections361 parameters17 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.41 e \u00c5\u22123\u0394\u03c1min = \u22120.34 e \u00c5\u22123\u0394\u03c1Absolute structure: Flack 1983, 3399 FrFlack parameter: 0.005 (5)SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812032928/bg2470Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S1600536812032928/bg2470Isup3.cmlSupplementary material file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Inter\u00admolecular C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings [centroid\u2013centroid distance = 3.745\u2005(3)\u2005\u00c5] are present in the crystal.In the title compound, [CoCl DOI: Click here for additional data file.10.1107/S1600536812041116/hy2591Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The bridging function of the Br atoms leads to a chain structure along [100]. Inter\u00adchain C\u2014H\u22efBr hydrogen bonds and \u03c0\u2013\u03c0 contacts between the thia\u00adzole rings [centroid\u2013centroid distances = 3.810\u2005(5) and 3.679\u2005(5)\u2005\u00c5] are observed.In the title coordination polymer, [CdBr DOI: 10.1107/S1600536811020861/hy2437Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, C\u2014H\u22efBr hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings [centroid\u2013centroid distance = 3.763\u2005(5)\u2005\u00c5] are present.In the title compound, [CdBr DOI: 10.1107/S1600536812033648/hy2576Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 32.12\u2005(14)\u00b0, while the pyridine ring and the benzene ring are oriented at a dihedral angle of 82.02\u2005(5)\u00b0. The coordinating water mol\u00adecule links with the carboxyl\u00adate group via an intra\u00admolecular O\u2014H\u22efO hydrogen bond. In the crystal, N\u2014H\u22efO, O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular network.In the title complex, [Cu(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.021wR(F2) = 0.058S = 1.133531 reflections203 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.97 e \u00c5\u22123\u0394\u03c1min = \u22120.50 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812034587/xu5599Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CuII atom is located on an inversion centre and has an overall octa\u00adhedral coordination environment. Two N and two O atoms from (pyzdc)2\u2212 ligands define the equatorial plane and two water mol\u00adecules are in axial positions, resulting in a typical tetra\u00adgonally Jahn\u2013Teller-distorted environment. Extensive classical O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efO and non-classical C\u2014H\u22efO hydrogen bonds, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions between aromatic rings of the cations [centroid\u2013centroid distance = 3.58\u2005(9)\u2005\u00c5], lead to the formation of a three-dimensional supra\u00admolecular structure.The title compound, (C DOI: 10.1107/S1600536811008981/wm2462Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordin\u00adation sphere about one CoII atom comprises four O-atom donors from two bidentate chelate (Ophenolate and Ocarbox\u00adyl) and bridging dianionic ligands and two water mol\u00adecules [Co\u2014O range = 2.0249\u2005(11)\u20132.1386\u2005(14)\u2005\u00c5], while that about the second CoII atom has four water mol\u00adecules and two bridging carboxyl\u00adate O-donor atoms [Co\u2014O range = 2.0690\u2005(14)\u20132.1364\u2005(11)\u2005\u00c5]. The coordinated water mol\u00adecules as well as the water mol\u00adecules of solvation give O\u2014H\u22efO water\u2013water and water\u2013carboxyl hydrogen-bonding inter\u00adactions in the three-dimensional framework structure.In polymeric title compound, {[Co II, see: Deng et al. 2(H2O)6]\u00b72H2O = 0.023 wR(F 2) = 0.061 S = 1.07 2560 reflections225 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.32 e \u00c5\u22123 \u0394\u03c1min = \u22120.47 e \u00c5\u22123 \u0394\u03c1 CrysAlis PRO used to solve structure: SIR92 (Altomare et al., 1994SHELXL97 (Sheldrick, 2008PLATON (Spek, 2009PLATON.Data collection: 10.1107/S1600536810052694/rn2077sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536810052694/rn2077Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The MnII ion exhibits a centrosymmetric octa\u00adhedral geometry involving two carboxyl\u00adate O atoms of two different pztmb ligands and four O atoms of four coordinated water mol\u00adecules. The packing shows a three-dimensional supra\u00admolecular network via O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.884\u2005(8) and 4.034\u2005(8)\u2005\u00c5] between the benzene ring of one pztmb anion and the pyrazine ring of an adjacent anion.The title compound, [Mn(C DOI: 10.1107/S160053681004506X/pk2282Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The full dimer is generated by an inversion center. In the dimer, the two IrIII atoms and two bridging Cl atoms form a perfectly planar ring. The two IrIII atoms and the two terminal Cl atoms also form a rigorous plane that is orthogonal [89.48\u2005(3)\u00b0] to the Ir2Cl2 ring. The plane of the cyclo\u00adpenta\u00addienyl ligand forms a dihedral angle of 54.06\u2005(7)\u00b0 with respect to the Ir2Cl2 ring.The asymmetric unit of the title complex, [Ir For the al. 2011.2Cl4(C12H19)2] = 0.019wR(F2) = 0.040S = 1.064440 reflections141 parametersH-atom parameters constrainedmax = 1.02 e \u00c5\u22123\u0394\u03c1min = \u22121.18 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536813005072/pk2466Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536813005072/pk2466Isup3.cmlSupplementary material file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atom in the title compound, [RuCl2(C15H10)(C18H15P)2]\u00b72C4H8O, has a distorted square-pyramidal conformation. The P and Cl atoms are at the base of the pyramid and the Ru\u2014Cindenyl\u00adidene bond is in the axial position. The two Cl ligands and the two phosphane ligands are in trans positions. The Cl\u2014Ru\u2014Cl and P\u2014Ru\u2014P angles are 157.71\u2005(2) and 166.83\u2005(2)\u00b0, respectively. The two independent tetra\u00adhydro\u00adfuran (THF) solvent mol\u00adecules are disordered. One THF mol\u00adecule was refined using a split-atom model. The second THF mol\u00adecule was accounted for by using program PLATON/SQUEEZE . The molecular conformation shows three intramolecular C\u2014H\u22efCl contacts and two C\u2014H\u22ef\u03c0 interactions while the crystal packing features an intermolecular C\u2014H\u22efCl contact and two very weak intermolecular C\u2014H\u22ef\u03c0 contacts.The RuSpek 2009. Acta Cr DOI: 10.1107/S1600536811016692/si2353Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Si2Cp centroid\u2013titanium distance is 2.0763\u2005(10)\u2005\u00c5 and the SiCp centroid\u2013titanium distance is 2.0793\u2005(10)\u2005\u00c5. The angle subtended at the Ti atom by the centroids of both cyclo\u00adpenta\u00addienyl rings is 131.22\u2005(4)\u00b0 and the Cl\u2014Ti\u2014Cl angle is 94.14\u2005(2)\u00b0.In the title compound, [Ti(C DOI: 10.1107/S1600536811046228/fj2469Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two heterocyclic rings of the 5-(2-pyridin-2-\u00adyl)pyrazine-2-carbonitrile ligand are almost coplanar [dihedral angle = 5.63\u2005(8)\u00b0], and the two chelating ligands are in an anti relationship. The mononuclear units are inter\u00adconnected along [010] through C\u2014H\u22efO(nitrate) and C\u2014H\u22efN(cyano) inter\u00adactions, forming an infinite chain. The mononuclear units are stacked along the a axis and inter\u00adconnected via inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions between adjacent pyridine and pyrazine rings [centroid\u2013centroid distances = 3.984\u2005(2) and 3.595\u2005(3)\u2005\u00c5], thus forming a three-dimensional supra\u00admolecular structure.In the mononuclear title complex, [Ag(NO DOI: 10.1107/S1600536811049221/zq2135Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "A significant Jahn\u2013Teller distortion is observed with two axial Cu\u2014O distances significantly longer than those in the equatorial CuO2N2 plane. In the crystal, \u03c0\u2013\u03c0 stacking inter\u00adactions, with centroid\u2013centroid distances of 3.547\u2005(3) or 3.728\u2005(3)\u2005\u00c5 between the phenanthroline rings, form layers parallel to (011).In the title compound, [Cu(C DOI: 10.1107/S1600536811018162/sj5144Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One of the N-bound H atoms is involved in an intra\u00admolecular N\u2014H\u22efCl hydrogen bond, while another one inter\u00adacts with the solvent methanol mol\u00adecule via an N\u2014H\u22efO hydrogen bond. Inter\u00admolecular O\u2014H\u22efCl and O\u2014H\u22efO hydrogen bonds link two further complex mol\u00adecules and four solvent mol\u00adecules into a centrosymmetric structural unit. The short distance of 3.624\u2005(4)\u2005\u00c5 between the centroids of the five- and the six-membered rings of two benzimidazole fragments indicates the presence of \u03c0\u2013\u03c0 inter\u00adactions.In the title complex, [CuCl(C I complexes with triphenlyphosphine ligands, see: Gennari et al. -thione ligand, see: Schneider et al. (C18H15P)2]\u00b72CH4O = 0.086wR(F2) = 0.217S = 1.077835 reflections510 parametersH-atom parameters constrainedmax = 1.53 e \u00c5\u22123\u0394\u03c1min = \u22120.57 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536814001251/cv5442Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814001251/cv5442Isup3.cdxSupporting information file. DOI: CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the complex mol\u00adecules are connected by O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 interactions between the benzene rings [centroid\u2013centroid distance = 3.8924\u2005(17)\u2005\u00c5] into a chain along [010]. Between adjacent chains, \u03c0\u2013\u03c0 inter\u00adactions occur between the pyridine rings [centroid\u2013centroid distance = 3.898\u2005(2)\u2005\u00c5], giving an overall two-dimensional architecture.In the title compound, [Co(C DOI: Click here for additional data file.10.1107/S1600536813006752/hy2620Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dipic and acr ligands are nearly planar [maximum deviation = 0.069\u2005(3)\u2005\u00c5 in dipic and 0.091\u2005(4)\u2005\u00c5 in acr] and the dihedral angle between their mean planes is 58.67\u2005(7)\u00b0. The Pd\u2014O bond lengths are nearly equal, but the Pd\u2014N bond lengths are slightly different. There is a short C\u2014H\u22efO inter\u00adaction in the mol\u00adecule involving the two ligands. In the crystal, complex mol\u00adecules are linked through C\u2014H\u22efO inter\u00adactions, forming a three-dimensional network. There are also a number of inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions present, the shortest ring centroid\u2013centroid distance being 3.622\u2005(3)\u2005\u00c5.In the title complex, [Pd(C II complex [Pt(C7H3NO4)(C13H9N)], see: Ha (C13H9N)] = 0.041wR(F2) = 0.100S = 0.993152 reflections244 parametersH-atom parameters constrainedmax = 1.21 e \u00c5\u22123\u0394\u03c1min = \u22121.14 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812011385/su2392Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "One of the Cp rings is substituted by a nitro\u00adbenzenamine group, which is essentially perpendicular to the substituted cyclo\u00adpenta\u00addienyl ring, with an N\u2014C(H2)\u2014C\u2014C torsion angle of 89.8\u2005(2)\u00b0. Intra\u00admolecular N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds occur. In the crystal, weak C\u2014H\u22efO hydrogen bonds link adjacent mol\u00adecules. In the title compound, [Fe(C DOI: 10.1107/S1600536812039177/zq2180Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The mononuclear units are inter\u00adconnected through \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.801\u2005(2) and 3.979\u2005(3)\u2005\u00c5] and the hexa\u00adfluoridophosphate anions are embedded within the inter\u00adstices. C\u00a0N\u22ef\u03c0 inter\u00adactions [N\u22efcentroid = 3.519\u2005(2)\u2005\u00c5] and C\u2014H.\u22efN hydrogen-bonding inter\u00adactions also occur.In the mononuclear title complex, [Ag(C DOI: 10.1107/S1600536811051403/bt5734Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the mol\u00adecules inter\u00adact through inter\u00admolecular N\u2014H\u22efO hydrogen bonds between the amino and meth\u00adoxy groups on the naphthalene ring systems and N\u2014H\u22ef\u03c0 inter\u00adactions between the amino groups and the naphthalene rings. Furthermore, weak C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the benzene rings are observed. The centroid\u2013centroid and inter\u00adplanar distances between the benzene rings of the aroyl group and the naphthalene ring systems of adjacent mol\u00adecules are 3.6954\u2005(8) and 3.2375\u2005(5)\u2005\u00c5, respectively. The dihedral angle between the mean planes of the benzene ring and the naphthalene ring system is 83.59\u2005(5)\u00b0. The benzene ring and the carbonyl group in the benzoyl unit are almost coplanar [C\u2014C\u2014C\u2014O torsion angle = 175.91\u2005(10)\u00b0].The title compound {systematic name: (4-aminophenyl)methanone}, C DOI: 10.1107/S1600536810041346/rz2496Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The O atom, the aldehyde H atom and the benzene ring of one of the FB anions are disordered over two positions. The O atoms were freely refined [refined occupancy ratio 0.79\u2005(2):0.21\u2005(2)], while the aldehyde H atoms and the benzene ring atoms were refined with fixed occupancy ratios of 0.8:0.2 and 0.5:0.5, respectively. In the ordered FB anion, the carboxyl\u00adate group is twisted away from the attached benzene ring (A) by 22.7\u2005(8)\u00b0. In the disordered FB anion, the corresponding angles are 15.6\u2005(10) and 11.4\u2005(11)\u00b0 for rings B and B\u2032, respectively. Benzene rings A and B are oriented at a dihedral angle of 24.2\u2005(7), A and B\u2032 at 43.0\u2005(8)\u00b0. The pyrazine ring makes dihedral angles of 67.5\u2005(4), 89.6\u2005(7) and 86.2\u2005(7)\u00b0, respectively, with benzene rings A, B and B\u2032. The pyrazine ligands bridge the CdII cations, forming polymeric chains running along the b-axis direction. In the crystal, O\u2014Hwater \u22ef Ocarboxyl\u00adate hydrogen bonds link adjacent chains into layers parallel to the bc plane. These layers are linked via C\u2014Hpyrazine \u22ef Oform\u00adyl hydrogen bonds, forming a three-dimensional network. \u03c0\u2013\u03c0 interactions [centroid\u2013centroid distances = 3.870\u2005(11)\u20133.951\u2005(5)\u2005\u00c5] further stabilize the crystal structure. There is also a weak C\u2014H\u22ef\u03c0 inter\u00adaction present.The polymeric title compound, [Cd(C DOI: 10.1107/S1600536813035010/su2679Isup2.hklStructure factors: contains datablock(s) I. DOI: CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal packing features \u03c0\u2013\u03c0 contacts between the pyridine rings of adjacent mol\u00adecules [centroid\u2013centroid distance = 3.773\u2005(5)\u2005\u00c5] and also between a pyridine ring of one mol\u00adecule and the five-membered chelate ring of an adjacent mol\u00adecule [centroid\u2013centroid distance = 3.668\u2005(4)\u2005\u00c5].In the title complex, [HgI DOI: 10.1107/S1600536811004041/ya2136Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The geometry of the resulting CuN2O4 coordination can be described as distorted octa\u00adhedral. In the crystal, there are several inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. An intra\u00admolecular N\u2014H\u22efO hydrogen bond occurs in one of the cations. Considerable \u03c0\u2013\u03c0 stacking inter\u00adactions are also observed between the aromatic rings of the cations, with centroid\u2013centroid distances of 3.4567\u2005(13), 3.5342\u2005(14), 3.6941\u2005(14) and 3.4568\u2005(13)\u2005\u00c5. These non-covalent inter\u00adactions connect the components, forming a three-dimensional supra\u00admolecular structure.The title compound, (C DOI: 10.1107/S160053681100674X/vm2078Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The PMe3 ligand is coordinated in the basal position, roughly cis to the Fe\u2014Fe bond. The Fe\u2014Fe distance of 2.4970\u2005(6)\u2005\u00c5 is relatively short compared to those (ca 2.53\u2005\u00c5) found in another monosubstituted diiron compound. A rigid planar dithiol\u00adate bridge is featured, with an angle of 2.78\u2005(1)\u00b0 between the Fe\u2014Fe bond and the normal to the pyrazine-2,3-dithiol\u00adate plane.In the title compound, [Fe DOI: 10.1107/S1600536811046770/hy2484Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, C\u2014H\u22efN, C\u2014H\u22efO and C\u2014H\u22efS inter\u00adactions link adjacent mol\u00adecules into layers parallel to the ac plane. A weak inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adaction occurs between the aromatic rings with a centroid\u2013centroid distance of 3.9412\u2005(9)\u2005\u00c5.In the title compound, [Cu(NCS) DOI: 10.1107/S1600536810050889/is2639Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The NiII atom exhibits a distorted octa\u00adhedral N3O3 environment. O\u2014H\u22efO hydrogen bonding between coordinated water and carboxyl\u00adate O atoms, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions [inter\u00adplanar distances between phen rings = 3.293\u2005(2)\u2005\u00c5] lead to a supermolecular assembly.The title compound, [Ni(C DOI: 10.1107/S1600536811026055/pv2419Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Characteristic features of the Cu6S6 skeleton are a total of six chemically identical \u03bc3-thiol\u00adate bridges and almost planar Cu2S2 units with a maximum deviation of 0.110\u2005(1)\u2005\u00c5 from the best plane. Each Cu2S2 unit then shares common Cu\u2013S edges with a neighbouring unit; the enclosed dihedral angle is 60.14\u2005(2)\u00b0. The geometric centre of the Cu6S6 cation lies on a crystallographic inversion centre. Cu\u2014S bond lengths range from 2.294\u2005(1) to 2.457\u2005(1)\u2005\u00c5, Cu\u2014N bond lengths from 2.005\u2005(3) to 2.018\u2005(3)\u2005\u00c5 and the non-bonding Cu\u22efCu distances from 2.5743\u2005(7) to 2.5892\u2005(6)\u2005\u00c5. C\u2014H\u22efF hydrogen-bond inter\u00adactions occur between the PF6\u2212 anion and the complex mol\u00adecule and between the PF6\u2212 anion and the acetonitrile solvent mol\u00adecule.The mol\u00adecular structure of the title compound, [Cu DOI: Click here for additional data file.10.1107/S1600536812050428/nr2037Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, dinuclear units with a Cd\u22efCd separation of 3.8208\u2005(7)\u2005\u00c5 are observed. Each of these dinuclear units is bridged via 3,3\u2032-bpda in a chelating/chelating and bridging fashion, generating a zigzag chain along the c axis. Neighboring chains are further packed via weak \u03c0\u2013\u03c0 inter\u00adactions between inter\u00adchain parallel 1,10-phen rings [centroid\u2013centroid distance = 3.5197\u2005(9)\u2005\u00c5] into a three-dimensional supra\u00admolecular architecture.In the title compound, [Cd(C DOI: 10.1107/S1600536811042085/im2322Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Of the two water H atoms, one is engaged in an intra\u00admolecular hydrogen bond with a free oxygen of the dianion whereas the second is engaged in an inter\u00admolecular hydrogen bond, building a corrugated layer parallel to (100). These layers are further connected through \u03c0\u2013\u03c0 stacking inter\u00adactions involving symmetry-related phenanthroline rings [centroid\u2013centroid distance = 3.5599\u2005(17) and 3.5617\u2005(18)\u2005\u00c5], building a three dimensionnal network. C\u2014H\u22ef\u03c0 inter\u00adactions involving the phenanthroline ring system are also observed.In the centrosymmetric dinuclear title complex, [Cu DOI: 10.1107/S1600536811007938/dn2661Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "I atom in the title centrosymmetric dinuclear compound, [Ag2(C11H10N3O)2(C11H11N3O)2]\u00b72H2O, shows a T-shaped coordination arising from bonding to the N atom of a neutral 2-[(pyrimidin-2-yl)amino\u00admeth\u00adyl]phenol ligand, the N atom of the 2-[(pyrimidin-2-yl)amino\u00admeth\u00adyl]phenolate anion [N\u2014Ag\u2014N = 171.8\u2005(1)\u00b0] and the terminal O atom of the other anion [Ag\u2014O = 2.606\u2005(3)\u2005\u00c5]. A pair of 2-[(pyrimidin-2-yl)amino\u00admeth\u00adyl]phenolate anions link the two AgI atoms to form the dinuclear compound. In the crystal, adjacent dinuclear mol\u00adecules are linked to the lattice water mol\u00adecules, generating an O\u2014H\u22efO- and N\u2014H\u22efO-connected three-dimensional network. In the crystal, the hy\u00addroxy H atom is disordered over two positions in a 1:1 ratio; one half-occupancy H atom is connected to one hy\u00addroxy group, whereas the other half-occupancy H atom is connected to another hy\u00addroxy group. The Ag DOI: Click here for additional data file.10.1107/S1600536812045783/xu5644Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The crystal packing is dominated by inter\u00admolecular C\u2014H\u22efF hydrogen bonds, C\u2014F\u22ef\u03c0 inter\u00adactions between the penta\u00adfluoro\u00adbenzene groups [F\u22efcentroid distances = 3.882\u2005(2) and 3.884\u2005(2)\u2005\u00c5] and \u03c0\u2013\u03c0 inter\u00adactions between the penta\u00adfluoro\u00adbenzene and cyclo\u00adpenta\u00addienyl rings [centroid\u2013centroid distance = 3.741\u2005(1)\u2005\u00c5].The mol\u00adecular structure of the title compound, [Fe(C DOI: 10.1107/S1600536810026772/hy2327Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the packing of the mol\u00adecules is controlled by C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions and weak Br\u22efBr halogen bonds [3.508\u2005(4)\u2005\u00c5], linking the mol\u00adecules into an infinite three-dimensional supra\u00admolecular network.In the title complex, [Cu(C DOI: Click here for additional data file.10.1107/S1600536813009847/zl2543Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the cation, the pyridinium rings attached to the central 1-aza\u00adniumyl-2-hy\u00addroxy\u00adethane fragment have an anti conformation, as indicated by the central C\u2014C\u2014C\u2014C torsion angle of \u2212166.5\u2005(6)\u00b0, and they are inclined to one another by 63.5\u2005(4)\u00b0. In the crystal, the cations and anions are linked through N\u2014H\u22efCl and O\u2014H\u22efCl hydrogen bonds. There are also \u03c0\u2013\u03c0 contacts [centroid\u2013centroid distances = 3.671\u2005(4) and 3.851\u2005(4)\u2005\u00c5] and a number of C\u2014H\u22efCl inter\u00adactions present, consolidating the formation of a three-dimensional supra\u00admolecular structure.The title compound, (C DOI: Click here for additional data file.10.1107/S160053681300425X/su2560Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The PbN2S4 coordination geometry approximates to a penta\u00adgonal bipyramid with one equatorial site vacant. The N atoms occupy the axial sites. One of the pyridine mol\u00adecules is disordered over two sets of sites in a 0.907\u2005(7):0.093\u2005(7) ratio and one of the tert-butyl groups is disordered over two sets of sites in a 0.534\u2005(6):0.466\u2005(6) ratio. An intra\u00admolecular C\u2014H\u22efO inter\u00adaction occurs in one of the ligands. In the crystal, pairs of short Pb\u22efS contacts [3.4018\u2005(11)\u2005\u00c5] generate a centrosymmetric dimeric assembly with the distant S atom lying in the region of the vacant coordination site of the metal atom. No directional packing inter\u00adactions occur.In the title compound, [Pb(C For the preparation of the ligand, see: Li & Xie 1997. For van20H26O2PS2)2(C5H5N)2] = 0.033wR(F2) = 0.068S = 1.0210953 reflections663 parameters572 restraintsH-atom parameters constrainedmax = 0.80 e \u00c5\u22123\u0394\u03c1min = \u22120.69 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I. DOI: 10.1107/S1600536813023945/hb7129Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The mol\u00adecular structure exhibits an S(6) ring motif, owing to an intra\u00admolecular O\u2014H\u22efO hydrogen bond. In the crystal, weak C\u2014H\u22efO contacts generate an infinite chain along the c axis. There are also \u03c0\u2013\u03c0 stacking inter\u00adactions between neighbouring isochromanedione benzene rings, with a centroid\u2013centroid distance of 3.755\u2005(1)\u2005\u00c5, and C\u2014O\u22ef\u03c0 inter\u00adactions with an O\u22efcentroid distance of 3.964\u2005(2)\u2005\u00c5.In the title compound, C DOI: 10.1107/S1600536811050975/fj2488Isup2.hkl Structure factors: contains datablock(s) I. DOI: 10.1107/S1600536811050975/fj2488Isup3.cml Supplementary material file. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "I cations, three p-toluene\u00adsulfonate (pts) anions, three hexa\u00admethyl\u00adene\u00adtetra\u00admine (hmt) mol\u00adecules and four water mol\u00adecules in the asymmetric unit of the title coordination polymer, {[Ag3(C7H7O3S)3(C6H12N4)3(H2O)]\u00b73H2O}n. Two of the pts anions show positional disorder of their O atoms in 0.60:0.40 and 0.50:0.50 ratios. The AgI ion is coordinated by three hmt mol\u00adecules in an approximate trigonal\u2013planar AgN3 arrangement. In each case, longer Ag\u2014O bonds to a water mol\u00adecule and a pts anion complete a distorted trigonal\u2013bipyramidal AgN3O2 geometry for the metal ion. In the crystal, the bridging hmt mol\u00adecules and pts ions generate a wave-like layer parallel to (001) and O\u2014H\u22efO hydrogen-bonding inter\u00adactions consolidate the packing.There are three Ag DOI: 10.1107/S1600536810048567/hb5739Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The central Cu2N2 core is a rhombus as the \u03bc2-azide ligands bridge in an asymmetric fashion. Each CuII atom is also coordinated by a monoanionic tridentate Schiff base ligand via the anti\u00adcipated oxide O, imine N and amine N atoms. The resulting N4O coordination geometry is based on a square pyramid. No specific inter\u00admolecular inter\u00adactions are noted in the crystal packing, but the amine H atoms form intra\u00admolecular N\u2014H\u22efO(oxide)/N(azide) hydrogen bonds.The complete mol\u00adecule of the title complex, [Cu For a r al. 2012. For add al. 1984.2(C13H19N2O2)2(N3)2] = 0.039wR(F2) = 0.105S = 1.063314 reflections198 parameters2 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.54 e \u00c5\u22123\u0394\u03c1min = \u22120.52 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812028954/sj5249Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The guanidinium cation (the C atom has site symmetry 3) and the octa\u00adhedral hexa\u00adaqua\u00adzinc(II) dication (the Zn2+ cation has site symmetry -3) are occupationally disordered in a 1.30:0.35 ratio. In the crystal, the components are linked by O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds to generate infinite (001) sheets. Weak aromatic \u03c0\u2013\u03c0 stacking [centroid\u2013centroid distance = 3.797\u2005(8)\u2005\u00c5] is also observed in the crystal.In the title mol\u00adecular salt, (CH DOI: 10.1107/S160053681202987X/hb6768Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angle between the nearly planar PtI2NS unit [maximum deviation = 0.083\u2005(2)\u2005\u00c5] and the acridine ligand [maximum deviation = 0.038\u2005(6)\u2005\u00c5] is 89.29\u2005(7)\u00b0. In the crystal structure, the complex mol\u00adecules are arranged in a V-shaped packing pattern along the c axis and linked by inter\u00admolecular C\u2014H\u22efO contacts into supra\u00admolecular chains. There are also several inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the six-membered rings, with a shortest ring centroid\u2013centroid distance of 3.804\u2005(5)\u2005\u00c5.In the title complex, [PtI DOI: 10.1107/S1600536810031387/tk2697Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In complex A, [Cd(C7H5O3)2(C6H6NO)2(H2O)2], the CdII cation is located on an inversion center and is coordinated by two salicylate anions, two nicotinamide (NA) ligands and two water mol\u00adecules in a slightly distorted octa\u00adhedral geometry. In complex B, [Cd(C7H5O3)2(C6H6NO)(H2O)2], the CdII cation is coordinated by two salicylate anions, one nicotinamide (NA) ligand and two water mol\u00adecules in an irregular seven-coordinate geometry. There are extensive intra\u00admolecular O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds as well as extensive inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonding in the crystal structure. \u03c0\u2013\u03c0 stacking between the pyridine and benzene rings, between the benzene rings, between the benzene and pyridine rings and between the pyridine rings further stabilize the crystal structure. A weak N\u2014H\u22ef\u03c0 inter\u00adaction also occurs. One of the lattice water mol\u00adecules is disordered over two positions with an occupancy ratio of 0.70:0.30.The crystal structure of the title compound, [Cd(C DOI: Click here for additional data file.10.1107/S1600536813006168/xu5682Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The chain in (I) extends parallel to [100] and is severely puckered, with a Zn\u22efZn distance of 8.3599\u2005(3)\u2005\u00c5 and a Zn\u22efZn\u22efZn angle of 107.516\u2005(3)\u00b0, as a result of hydrogen-bonding inter\u00adactions of the types O\u2014H\u22efO and C\u2014H\u22efO. There is no evidence for \u03c0\u2013\u03c0 inter\u00adactions in (I). The differences between the solvent-free and solvent-containing structures can be accounted for by the absence of the ethanol solvent mol\u00adecule and the use of the converging pair of O atoms in the bis-monodentate bridging HBTC2\u2212 ligand in (I).The title one-dimensional coordination polymer, [Zn(C al. 1997. Chem. M For a r al. 2009. 9H4O6)(C5H5N)2] = 0.028wR(F2) = 0.075S = 1.034402 reflections257 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.32 e \u00c5\u22123\u0394\u03c1min = \u22120.27 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536813014347/cq2004Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II complex, [Cu(C7H4BrO2)2(C10H14N2O)2(H2O)2], contains two 4-bromo\u00adbenzoate (PBB), two diethyl\u00adnicotinamide (DENA) monodentate ligands and two water mol\u00adecules. The four O atoms in the equatorial plane around the CuII ion form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by two N atoms of the DENA ligands in the axial positions. Intra\u00admolecular O\u2014H\u22efO hydrogen bonds link the water mol\u00adecules to the carboxyl\u00adate groups. The dihedral angles between the carboxyl\u00adate groups and the adjacent benzene rings are 3.1\u2005(3) and 3.74\u2005(17)\u00b0, while the pyridine rings and the benzene rings are oriented at dihedral angles of 6.81\u2005(10) and 3.38\u2005(12)\u00b0. In the crystal, inter\u00admolecular O\u2014H\u22efO hydrogen bonds link the mol\u00adecules into double chains along the b axis. C\u2014H\u22efO inter\u00adactions are also observed. \u03c0\u2013\u03c0 contacts between pyridine rings [centroid\u2013centroid distance = 3.485\u2005(2)\u2005\u00c5] may further stabilize the crystal structure.The title Cu N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C10H14N2O)2(H2O)2] = 0.035 wR(F 2) = 0.079 S = 1.03 7011 reflections463 parameters5 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 1.08 e \u00c5\u22123 \u0394\u03c1min = \u22120.59 e \u00c5\u22123 \u0394\u03c1Absolute structure: Flack 1983, 2353 FrFlack parameter: 0.412 (7) APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811034787/su2303Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The NH2CH2CHBrCH2Br ligand contains a chiral carbon atom. The Fe\u2014N bond length is 2.011\u2005(3)\u2005\u00c5 and the Fe\u2014Cp centroid distance is 1.7189\u2005(5)\u2005\u00c5. In the crystal, the ions are linked via two N\u2014H\u22efF inter\u00adactions and a weak N\u2014H\u22efBr inter\u00adaction.The title compound, [Fe(\u03b7 DOI: 10.1107/S1600536812025925/fj2567Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The water mol\u00adecule at the apical position shows a long bond [Cu\u2014O = 2.276\u2005(2)\u2005\u00c5]. The basal plane is formed by two N atoms of the 2,2\u2032-bipyridine ligand and two carboxyl\u00adate O atoms from a malonate group. The five-membered chelate ring is almost planar [maximum deviation = \u22120.006\u2005(2)\u2005\u00c5], while the six-membered chelate ring defined by the malonate ligand adopts a distorted boat conformation. In the crystal, CuII complex mol\u00adecules and lattice water mol\u00adecules are connected by O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. The crystal packing is further stabilized by \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.563\u2005(2)\u20133.828\u2005(2)\u2005\u00c5].In the title compound, [Cu(C For rel al. 1998; Cui et al. 2005. For rin al. 2005.4H4O4)(C10H8N2)(H2O)]\u00b72H2O = 0.049wR(F2) = 0.119S = 1.053782 reflections242 parameters6 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.97 e \u00c5\u22123\u0394\u03c1min = \u22120.52 e \u00c5\u22123\u0394\u03c1CrysAlis CCD used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812024889/bt5931Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The mean bond lengths of the two Ru complexes are Ru\u2014C = 2.276\u2005\u00c5, Ru\u2014P\u00a0= 2.3816\u2005\u00c5, and Ru\u2014Cl = 2.3924\u2005\u00c5. Both chloride ligands form only intra\u00admolecular C\u2014H\u22efCl inter\u00adactions. Seven weak inter\u00admolecular C\u2014H\u22efF inter\u00adactions involving mainly arene H atoms consolidate the crystal packing, which reveals an approximate c/2 pseudo-translation relating the two independent Ru complex mol\u00adecules.The asymmetric unit of the title compound, [FeRuCl(C DOI: 10.1107/S1600536811036506/kp2346Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The complex mol\u00adecules are linked by inter\u00admolecular O\u2014H\u22efO hydrogen bonds and partly overlapping \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 4.017\u2005(2)\u2005\u00c5] into a three-dimensional supra\u00admolecular network.In the title compound, [Zn(C DOI: 10.1107/S1600536811019519/hy2433Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal packing is stabilized by N\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.576\u2005(5)\u2005\u00c5].The title salt, (C DOI: 10.1107/S1600536811024792/aa2014Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, O\u2014H\u22efCl, C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 contacts between the pyridine rings [centroid\u2013centroid distances = 3.7236\u2005(17) and 3.6026\u2005(19)\u2005\u00c5] stabilize the structure. Intra\u00admolecular C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds are also present.In the title compound, [VClO(C DOI: 10.1107/S1600536812040251/hy2588Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The ZnII ion shows a distorted octa\u00adhedral coordination geometry and is coordin\u00adated by two N atoms from two SCN\u2212 anions and four N atoms from two organic ligands. The L ligands act as bridging bis-chelating ligands with cis coordination modes at the ZnII ion. One-dimensional coordination polymers are arranged into layers by \u03c0\u2013\u03c0 stacking inter\u00adactions between the imidazole rings of adjacent chains, with an inter\u00adplanar distance of 3.46\u2005(1)\u2005\u00c5 and centroid\u2013centroid distances of 3.8775\u2005(16)\u2005\u00c5. One of the thio\u00adcyanate ligands is disordered over two positions with an occupancy factor of 0.564\u2005(3) for the major component. The partially occupied water mol\u00adecule forms an O\u2014H\u22efS hydrogen bond with the disordered thio\u00adcyanate group.The title one-dimensional coordination polymer, {[Zn(NCS) For coo al. 2004. For sim al. 2002; Luan et al. 2006. For the al. 2008. 2(C24H20N6)2]\u00b70.28H2O = 0.038 wR(F 2) = 0.102 S = 1.04 4707 reflections362 parameters30 restraintsH-atom parameters constrainedmax = 0.38 e \u00c5\u22123 \u0394\u03c1min = \u22120.33 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008DIAMOND (Brandenburg & Putz, 2008SHELXL97.Data collection: 10.1107/S1600536810027571/gk2287sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810027571/gk2287Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Intra\u00admolecular C\u2014H\u22efO hydrogen bonds and inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings [centroid\u2013centroid distances = 3.637\u2005(4) and 3.818\u2005(4)\u2005\u00c5] are present in the crystal structure.In the title compound, [ZnI DOI: 10.1107/S1600536810046763/hy2375Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One AgI atom is coordinated by two pyridine N atoms from two symmetry-related A ligands in a geometry slightly distorted from linear [N\u2014Ag\u2014N = 173.2\u2005(3)\u00b0], forming a left-handed helical chain, while the other AgI atom is coordinated by two pyridine N atoms from two symmetry-related B ligands in a bent arrangement [N\u2014Ag\u2014N = 157.1\u2005(3)\u00b0], forming a right-handed helical chain. Both helical chains have the same pitch length [10.4007\u2005(7)\u2005\u00c5], propagate along the b-axis direction and are alternately arranged via Ag\u22efAg [3.0897\u2005(12)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.564\u2005(7) and 3.518\u2005(6)\u2005\u00c5], resulting in the formation of a two-dimensional supra\u00admolecular network extending parallel to the ab plane. Inter\u00admolecular N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efF hydrogen-bonding inter\u00adactions occur between the helical chains and the anions.In the asymmetric unit of the title polymeric complex, {[Ag(C I coordination polymers with dipyridyl ligands, see: Leong & Vittal ](CF3O3S) = 0.088wR(F2) = 0.238S = 1.095797 reflections415 parameters6 restraintsH-atom parameters constrainedmax = 2.70 e \u00c5\u22123\u0394\u03c1min = \u22121.89 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536813016309/sj5332Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "I ions resulted in the formation of the polymeric title compound, {[Ag(C10H8N2)](C17H9F6O4)}n, in which the metal atoms are bridged by the 4,4\u2032-bipyridine ligands, generating cationic chains extending along [010]. The dihedral angles between the benzene rings in the anion and the pyridine rings in the cation are 72.42\u2005(9) and 9.36\u2005(10)\u00b0, respectively. The mol\u00adecular conformation of the anion is stabilized by intra\u00admolecular C\u2014H\u22efF hydrogen bonds. In the crystal, the anions inter\u00adact with the cationic chains via C\u2014H\u22efO hydrogen bonds, forming layers parallel to (001), in which weak \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.975\u2005(3)\u20134.047\u2005(3)\u2005\u00c5] involving the pyridine rings of adjacent 4,4\u2032-bipyridine ligands are present. The planes are further assembled into a three-dimensional network by O\u2014H\u22efO hydrogen bonds.Assembly of the flexible dicarb\u00adoxy\u00adlic ligand 4-benzoate and 4,4\u2032-bipyridine as co-ligand with Ag DOI: 10.1107/S1600536812015322/rz2731Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II(C2H8N2)3]Cl2, was obtained unexpectedly as the product of an attempted solvothermal synthesis of cobalt selenide from the elements in the presence of NH4Cl in ethyl\u00adenedi\u00adamine solvent. The three chelate rings of the distorted octa\u00adhedral [Co(C2H8N2)3]2+ complex cation adopt twisted conformations about their C\u2014C bonds. The spread of cis-N\u2014Co\u2014N bond angles [80.17\u2005(6)\u201398.10\u2005(6)\u00b0] in the title compound is considerably greater than the equivalent data for [CoIII(C2H8N2)3]Cl3 . In the crystal, the components are linked by numerous N\u2014H\u22efCl hydrogen bonds, generating a three-dimensional network in which the cationic complexes are stacked in columns along [010] and separated by columns of chloride anions.The title compound, [Co al. 2008. Angew. III\u2013tris-ethyl\u00adenedi\u00adamine complex with chloride counter-anions has been reported by Takamizawa et al. 3]Cl2 = 0.024wR(F2) = 0.059S = 1.062700 reflections232 parametersAll H-atom parameters refinedmax = 0.26 e \u00c5\u22123\u0394\u03c1min = \u22120.37 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536813013135/hb7079Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The cyano-bound 4-cyano\u00adpyridine mol\u00adecule also is disordered across the inversion centre. The four N atoms of the pyrrole rings of the dianionic 5,10,15,20-tetra\u00adphenyl\u00adporphyrin ligand occupy the equatorial sites of the octa\u00adhedron [Mg\u2014N = 2.0552\u2005(10) and 2.0678\u2005(11)\u2005\u00c5] and the axial Mg\u2014 bond length is 2.3798\u2005(12)\u2005\u00c5. The crystal packing is stabilized by weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions.In the title complex, [Mg(C DOI: Click here for additional data file.10.1107/S1600536812049434/zs2246Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "It is built up from discrete [Zn{(CH3)2SO}6]2+ units connected through non-classical hydrogen bonds to linear I42\u2212 polyiodide anions (C\u2014H\u22efI = 3.168\u2005\u00c5). The ZnII ion in the cation has an octa\u00adhedral coordination geometry, with all six Zn\u2014O bond lengths being equivalent, at 2.111\u2005(4)\u2005\u00c5. The linear polyiodide anion contains a neutral I2 mol\u00adecule weakly coordinated to two iodide ions.The title compound, [Zn{(CH DOI: 10.1107/S1600536813028377/fj2643Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Each Mo2 4+ unit is equatorially coordinated by four pentafluoro\u00adbenzoate groups, while the axial positions are occupied by two 4,4\u2032-bipyridine mol\u00adecules. The Mo\u2014Mo bond length of 2.1227\u2005(4)\u2005\u00c5 is representative of a dimolybdenum quadruple bond. An infinite linear chain parallel to [110] is formed by the Mo2 4+ unit coordinating axially to the two N atoms of the 4,4\u2032-bipyridine ligand [Mo\u2014N = 2.594\u2005(2)\u2005\u00c5]. The crystal packing shows mol\u00adecules linked together into a three-dimensional network via Mo\u2014N coordination inter\u00adactions and weak \u03c0\u2013\u03c0 stacking inter\u00adactions between perfluoro\u00adphenyl rings [centroid\u2013centroid distance = 3.7280\u2005(3)\u2005\u00c5 and centroid-to-plane distance = 3.6103\u2005(12)\u2005\u00c5 between two penta\u00adfluoro\u00adphenyl rings]. In the title compound, [Mo DOI: 10.1107/S1600536811031734/jj2094Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The water mol\u00adecules and 3,5-bis\u00ad(trifluoro\u00admeth\u00adyl)pyrazolide anions act as bridges between the potassium cations. Each potassium cation is surrounded by four O atoms [K\u2014O = 2.705\u2005(3)\u20132.767\u2005(3)\u2005\u00c5] and four F atoms [K\u2014F = 2.870\u2005(7)\u20133.215\u2005(13)\u2005\u00c5]. The water mol\u00adecules and the 3,5-bis\u00ad(trifluoro\u00admeth\u00adyl)pyrazolide anions are connected by O\u2014H\u22efN hydrogen bonds, forming layers in the ab plane. All \u2013CF3 groups show rotational disorder between two orientations each.The asymmetric unit of the title compound, [K(C DOI: 10.1107/S1600536810027133/cv2744Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ReI atom is facially surrounded by three carbonyl ligands and a tridentate bis\u00ad(pyridin-2-ylmeth\u00adyl)amine ligand in a distorted octahedral environment. N\u2014H\u22efBr, O\u2014H\u22efBr, C\u2014H\u22efO and C\u2014H\u22efBr hydrogen bonds are present in the crystal structure and \u03c0\u2013\u03c0 stacking is also observed [centroid\u2013centroid distances = 3.669\u2005(1)\u2005\u00c5 and 4.054\u2005(1)\u2005\u00c5], giving rise to a three-dimentional network. The mol\u00adecules pack in a head-to-head fashion along the ac plane.The title compound, DOI: 10.1107/S1600536812019654/ru2033Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "All of the CuII atoms in the trinuclear centrosymmetric title complex are six-coordinated in a distorted octa\u00adhedral geometry with N2O4 and N4O2 environments for the outer and central CuII atoms, respectively. Various inter\u00adactions, including numerous O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014O\u22ef\u03c0 stacking of the pyridine and carboxyl\u00adate groups [O\u22efcentroid distances = 3.669\u2005(2) and 3.668\u2005(2)\u2005\u00c5] are observed in the crystal structure. The title compound, [Cu DOI: 10.1107/S1600536812022039/qm2066Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Weak C\u2014H\u22efN and C\u2014H\u22efS hydrogen bonds between the 1-ethyl-4,4\u2032-bipyridin-1-ium cations and mnt anions and weak \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings of the cations [centroid\u2013centroid distances = 3.808\u2005(3) and 3.972\u2005(3)\u2005\u00c5] lead to a two-dimensional network parallel to (010).In the anion of the title compound, (C DOI: 10.1107/S1600536813015493/hy2630Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angle between the benzene rings is 36.86\u2005(14)\u00b0. In the crystal, mol\u00adecules are linked into inversion dimers by pairs of weak C\u2014H\u22efO hydrogen bonds. In addition, \u03c0\u2013\u03c0 [centroid\u2013centroid distance = 3.7279\u2005(16)\u2005\u00c5] and weak C\u2014H\u22ef\u03c0 inter\u00adactions are observed.In the title Schiff base complex, [Cu(C DOI: 10.1107/S1600536812033491/lh5504Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The angles between the mean planes of the pyrrolo-indole ring and the phenyl\u00adsulfonyl and p-toluene\u00adsulfonyl rings are 73.7\u2005(6) and 80.6\u2005(0)\u00b0, respectively. The dihedral angle between the mean planes of the two benzene rings is 78.7\u2005(4)\u00b0. In the crystal, both classical N\u2014H\u22efO and non-classical C\u2014H\u22efO inter\u00admolecular hydrogen-bonding inter\u00adactions are observed, as well as weak \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.6258\u2005(8) and 3.9298\u2005(8)\u2005\u00c5], which contribute to the stability of the packing.The title compound, C DOI: 10.1107/S1600536810039425/fl2319Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the solid state, the second acidic proton of isophthalic acid is partially transferred to the imidazole N atom of the adeninium cation [refined O\u2014H versus N\u2014H ratio = 0.70\u2005(11):0.30\u2005(11)]. Through the partially transferred proton, the adeninium cation is strongly hydrogen bonded (N\u2014H\u22efO/O\u2014H\u22efN) to the isophthalate anion. This strong inter\u00adaction is assisted by another N\u2014H\u22efO hydrogen bond originating from the adeninium NH2 group towards the isophthalate keto O atom, with an R22(8) graph-set motif. This arrangement is linked via N\u2014H\u22efO hydrogen bonds to the O atoms of the carboxyl\u00adate group of an isophthalate anion. Together, these hydrogen bonds lead to the formation criss-cross zigzag isophthalate\u22efadeninium chains lying parallel to (501) and (50-1). The adeninium cations and the isophthalate anions are arranged in infinite \u03c0 stacks that extend along the c-axis direction [inter\u00adplanar distance = 3.305\u2005(3)\u2005\u00c5]. Mol\u00adecules are inclined with respect to this direction and within the stacks they are offset by ca. half a mol\u00adecule each. Combination of the N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds with the \u03c0\u2013\u03c0 inter\u00adactions forms infinitely stacked isophthalate\u22efadeninium chains, thus leading to a two-dimensional supra\u00admolecular structure with parallel inter\u00addigitating layers formed by the \u03c0 stacked isophthalate\u22efadeninium chains. The DMF mol\u00adecules of crystallization are bonded to the adeninium cations through strong N\u2014H\u22efO hydrogen bonds and project into the lattice space in between the anions and cations. There are also C\u2014H\u22efO hydrogen bonds present which, combined with the other inter\u00adactions, form a three-dimensional network. The crystal under investigation was found to be split and was handled as if non-merohedrally twinned.In the title proton-transfer organic salt, C \u00c5b = 46.05 (2) \u00c5c = 3.7832 (18) \u00c5V = 6674 (5) \u00c53Z = 16K\u03b1 radiationMo \u22121\u03bc = 0.11 mmT = 100 K0.55 \u00d7 0.15 \u00d7 0.04 mmBruker SMART APEX CCD diffractometerTmin = 0.730, Tmax = 1.000Absorption correction: multi-scan 2703 reflections with R[F2 > 2\u03c3(F2)] = 0.088wR(F2) = 0.233S = 1.053888 reflections251 parameters2 restraintsH atoms treated my a mixture of independent and constrained refinementmax = 0.58 e \u00c5\u22123\u0394\u03c1min = \u22120.50 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, New_Global_Publ_Block. DOI: 10.1107/S1600536813034971/su2677Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536813034971/su2677Isup3.cmlSupporting information file. DOI: CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Four water O atoms in the equatorial plane around the ZnII ion [Zn\u2014O = 2.089\u2005(2) and 2.128\u2005(2)\u2005\u00c5] form a slightly distorted square-planar arrangement and the distorted octa\u00adhedral geometry is completed by the two N atoms [Zn\u2014N = 2.117\u2005(2)\u2005\u00c5] from two isonicotinamide ligands. In the anion, the carboxyl\u00adate group is twisted from the attached benzene ring at 9.0\u2005(2)\u00b0. In the crystal, a three-dimensional hydrogen-bonding network, formed by classical O\u2014H\u22efO and N\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds, consolidates the crystal packing, which exhibits \u03c0\u2013\u03c0 stacking between the benzene and pyridine rings, with centroid\u2013centroid distances of 3.458\u2005(2) and 3.609\u2005(2)\u2005\u00c5. One of the two H atoms of each uncoordinating water mol\u00adecule is disordered over two orientations with an occupancy ratio of 0.60:0.40.The asymmetric unit of the title compound, [Zn(C DOI: Click here for additional data file.10.1107/S1600536813006466/xu5683Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, the components are linked by O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and weak aromatic \u03c0\u2013\u03c0 stacking [shortest centroid\u2013centroid separation = 3.778\u2005(2)\u2005\u00c5] inter\u00adactions. (001) layers of alternating organic cations and complex inorganic anions are apparent.In the title compound, (C DOI: 10.1107/S1600536814002530/hb7194Isup2.hklStructure factors: contains datablock(s) I. DOI: CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Fe atom and the phenolate ligand are disordered across the porphyrin ring with the two phenolates appearing to be roughly related by a center of symmetry. The occupancies of the two phenolate groups refined to 0.788\u2005(3) for the major component and 0.212\u2005(3) for the minor component. The structure shows extraordinary Fe displacements of 0.488\u2005(4) (major) and 0.673\u2005(4)\u2005\u00c5 (minor) from the 24-atom mean plane of the porphyrin. The Fe\u2014Np distances range from 2.063\u2005(4) to 2.187\u2005(6)\u2005\u00c5 and the Fe\u2014O distances are 1.903\u2005(5)\u2005\u00c5 for major component and 1.87\u2005(2)\u2005\u00c5 for minor component. The four phenyl groups attached to the porphyrin ring form dihedral angles of 63.4\u2005(4), 49.6\u2005(4), 62.4\u2005(4), and 63.3\u2005(4)\u00b0 with the three nearest C atoms of the porphyrin ring. The major and minor component phenolate groups form dihedral angles of 24.9\u2005(4)\u00b0 and 24.8\u2005(4)\u00b0, respectively, with the four porphyrin N atoms. The Fe\u22efFe distance between the two iron(III) atoms of adjacent porphyrin mol\u00adecules is 6.677\u2005(3)\u2005\u00c5. No close inter\u00admolecular inter\u00adaction was observed. The crystal studied was twinned by inversion, with a major\u2013minor component ratio of 0.53\u2005(3):0.47\u2005(3).The title compound, [Fe(C DOI: 10.1107/S160053681302607X/pk2492Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The structure is isotypic with [CdBr2(C4H5N3O)2] . There are two inter\u00adligand N\u2014H\u22efBr hydrogen bonds, generating two hydrogen-bonded rings stabilizing the coordination sphere. The complex aggregates, forming supra\u00admolecular chains, sheets and staircases through N\u2014H\u22efO and N\u2014H\u22efBr hydrogen bonding and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.616\u2005(2)\u2005\u00c5].In the title complex, [ZnBr DOI: 10.1107/S1600536810049305/hg2756Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, O\u2014H\u22efO, N\u2014H\u22efO and N\u2014H\u22efCl hydrogen bonds link the complex cations and nitrate anions into a three-dimensional network. \u03c0\u2013\u03c0 inter\u00adactions between the thia\u00adzole and imidazole rings and between the thia\u00adzole and benzene rings are observed [centroid\u2013centroid distances = 3.592\u2005(3) and 3.735\u2005(3)\u2005\u00c5].In the title compound, [NiCl(C DOI: 10.1107/S1600536812029728/hy2553Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, the complexes are linked into a three-dimensional supra\u00admolecular network by both O\u2014H\u22efO hydrogen-bonding inter\u00adactions between water mol\u00adecules and the uncoordinated carboxyl\u00adate O atoms of neighboring mol\u00adecules, and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions between neighboring phenanthroline rings with centroid\u2013centroid distances of 3.4654\u2005(17) and 3.697\u2005(2)\u2005\u00c5.In the title complex, [Zn(C DOI: 10.1107/S1600536810049573/vm2055Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the L ligand, the dihedral angle between benzoimidazole and benzotriazole ring systems is 10.8\u2005(3)\u00b0. In the crystal, the complex mol\u00adecules are connected by O\u2014H\u22efN hydrogen bonds; inter\u00admolecular \u03c0\u2013\u03c0 stacking is also observed [centroid\u2013centroid distances of 3.668\u2005(5)\u2005\u00c5 between triazole and benzene rings and 3.780\u2005(5)\u2005\u00c5 between imidazole rings].In the title complex, [Cd(C DOI: Click here for additional data file.10.1107/S1600536813001827/xu5670Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dimethyl sulfoxide solvent mol\u00adecules and CuII complex mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonding. In addition, C\u2014H\u22efO contacts and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.590\u2005(1)\u2005\u00c5] occur.In the title complex, [Cu(C DOI: 10.1107/S1600536811003424/bt5466Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two essentially planar 2,1,3-benzoselenadiazole ligands [maximum deviations = 0.012\u2005(2) and 0.030\u2005(2)\u2005\u00c5] are approximately coplanar [dihedral angle = 6.14\u2005(6)\u00b0]. In the crystal, short inter\u00admolecular Se\u22efBr, Se\u22efN and N\u22efN inter\u00adactions are observed. These short inter\u00adactions and inter\u00admolecular C\u2014H\u22efBr hydrogen bonds link the complex mol\u00adecules into two-dimensional arrays parallel to the ac plane.In the title complex, [CuBr DOI: 10.1107/S160053681005422X/sj5081Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The anion is an eight-coordinate complex with a square-anti\u00adprismatic geometry around the BiIII atom. In the crystal, extensive O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, as well as ion pairing, C=O\u22ef\u03c0 inter\u00adactions [O\u22efcentroid distance = 3.583\u2005(5)\u2005\u00c5], \u03c0\u2013\u03c0 stacking [centroid\u2013centroid distance = 3.864\u2005(3)\u2005\u00c5], and C\u2014H\u22ef\u03c0 and C\u2014H\u22efO inter\u00adactions, play an important role in the formation and stabilization of the three-dimensional supra\u00admolecular structure.The asymmetric unit of the ionic title compound, (C DOI: 10.1107/S160053681202630X/su2401Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "An intra\u00admolecular C\u2014H\u22efO hydrogen bond occurs. The crystal structure is stabilized by C\u2014H\u22efBr hydrogen bonds and \u03c0\u2013\u03c0 contacts between the pyridine rings [centroid\u2013centroid distances = 3.582\u2005(5) and 3.582\u2005(5)\u2005\u00c5].In the title compound, [CdBr DOI: 10.1107/S1600536812033168/hy2573Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The metal atom is in a distorted tetra\u00adhedral coordination environment with the two S atoms from two thio\u00adurea ligands and two O atoms from two acetate anions as the coordinating atoms. All H atoms of the thio\u00adurea ligands are involved in N\u2014H\u22efO and N\u2014H\u22efS hydrogen bonds, leading to a three-dimensional network.The title compound, [Co(CH II compound, see: Cavalca et al. 2(CH4N2S)2] = 0.016wR(F2) = 0.041S = 1.043087 reflections190 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.35 e \u00c5\u22123\u0394\u03c1min = \u22120.19 e \u00c5\u22123\u0394\u03c1APEX2 I, global. DOI: 10.1107/S1600536814002074/sj5387Isup2.hklStructure factors: contains datablock(s) I. DOI: CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The NiII atom exhibits a slightly distorted square-planar coordination geometry. In the crystal, the water mol\u00adecule links anions and cations into a three-dimensional network via O\u2014H\u22efN, O\u2014H\u22efS and O\u2014H\u22efO hydrogen bonds. The structure is further stabilized by weak S\u22ef\u03c0 contacts [S\u22efcentroid = 3.8047\u2005(9)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking inter\u00adactions [centriod\u2013centroid distance = 3.8653\u2005(7)\u2005\u00c5].In the title ion-pair complex, (C DOI: 10.1107/S1600536811045430/rz2659Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atom in the title complex, [Cd(NO3)(C18H40N4)]NO3\u00b70.5H2O, is coordinated within a cis-N4O2 donor set provided by the tetra\u00addentate macrocyclic ligand and two O atoms of a nitrate anion; the coordination geometry is distorted octa\u00adhedral. The lattice water mol\u00adecule is located on a twofold rotation axis. N\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22efO inter\u00adactions link the complex cations into a supra\u00admolecular layer in the bc plane. Layers are connected by O\u2014H\u22efO hydrogen bonds between the lattice water mol\u00adecule and the non-coordinating nitrate anion, as well as by weak C\u2014H\u22efO contacts.The Cd DOI: 10.1107/S160053681201238X/xu5483Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Pd atom is located on an inversion centre, and thus the asymmetric unit contains one half of the complex and the PdN2Cl2 unit is exactly planar. The dihedral angle between the PdN2Cl2 unit and the acridine ligand is 84.66\u2005(6)\u00b0. In the crystal, the complex mol\u00adecules are stacked in columns along the a axis connected by C\u2014H\u22efCl hydrogen bonds, forming chains along [110]. In the columns, numerous inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the six-membered rings are present, the shortest ring centroid\u2013centroid distance being 3.722\u2005(4)\u2005\u00c5.In the title complex, [PdCl X 2(acr)2] , see: Ha 2] = 0.059 wR(F 2) = 0.107 S = 0.98 2124 reflections142 parametersH-atom parameters constrainedmax = 0.83 e \u00c5\u22123 \u0394\u03c1min = \u22120.66 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811053335/bq2326Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The equatorial positions are occupied by two N atoms from two 2,2\u2032-bipyridyl ligands [Fe\u2014N = 2.121\u2005(5) and 2.147\u2005(5)\u2005\u00c5] and two Cl atoms [Fe\u2014Cl = 2.220\u2005(2) and 2.2074\u2005(18)\u2005\u00c5]. Weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions consolidate the crystal packing.In the crystal structure of the title compound, [FeCl DOI: 10.1107/S1600536811016035/zq2098Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The resulting CdN3O4 coordination polyhedron approximates to a very distorted penta\u00adgonal bipramid with one O and one N atom in axial positions. One of the ABTC ligands is bridging to an adjacent metal atom, generating an infinite chain propagating in [100]. A three-dimensional network is constructed from N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid separations = 3.641\u2005(2) and 3.682\u2005(3)\u2005\u00c5].In the title coordination polymer, [Cd(C DOI: 10.1107/S160053681201642X/hb6715Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Each CuI cation is in a distorted tetra\u00adhedral coordination environment defined by two iodide anions and two nitro\u00adgen atoms from two individual HIPy ligands. Two CuI atoms are connected by two HIPy ligands to form a dimer and these dimers are further bridged through the iodide atoms, leading to a chain structure extending parallel to [100]. Moreover, inter\u00admolecular N\u2014H\u22efI hydrogen bonds and weak \u03c0\u2013\u03c0 stacking inter\u00adactions between pyridyl rings link the chains into a two-dimensional supra\u00admolecular network in the ac plane.The title polymeric compound, [Cu N-heterocyclic carb\u00adoxy\u00adlic acid ligands, see: Chen & Tong 2] = 0.031 wR(F 2) = 0.080 S = 1.06 1682 reflections118 parametersH-atom parameters constrainedmax = 0.86 e \u00c5\u22123 \u0394\u03c1min = \u22120.69 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811033605/zl2401Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angle between the triazole and pyridine rings is 23.15\u2005(12)\u00b0. In the crystal, mol\u00adecules are linked by N\u2014H\u22efN and O\u2014H\u22efS hydrogen bonds. Offset \u03c0\u2013\u03c0 stacking between parallel pyridine rings of adjacent mol\u00adecules is also observed, the centroid\u2013centroid distance being 3.6319\u2005(14)\u2005\u00c5.In the title compound, [Cd(NCS) DOI: 10.1107/S1600536812032473/xu5596Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The anion bears two nine-coordinate CeIII ions, each with a distorted tricapped trigonal\u2013prismatic geometry. In the crystal, inter\u00admolecular C\u2014H\u22efO, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, as well as \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances of 3.514\u2005(3)\u2005\u00c5 connect the various components into a supra\u00admolecular structure.The title compound, {(C DOI: 10.1107/S1600536811004995/hy2405Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The O\u2014Al\u2014N bite angle of the chelating ligand is 94.14\u2005(9)\u00b0. The O\u2014C\u2014C\u2014C\u2014N backbone of the ligand is nearly coplanar (r.m.s. deviation = 0.029\u2005\u00c5) and the Al atom deviates significantly from the mean plane by 0.525\u2005(3)\u2005\u00c5. In the crystal, weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions are observed.The mol\u00adecular structure of the title compound, [Al(CH DOI: 10.1107/S1600536812005880/zq2142Isup3.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S1600536812005880/zq2142Isup4.cdxSupplementary material file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II complex, [Mn(C7H4BrO2)2(C10H14N2O)2(H2O)2], the MnII cation is located on an inversion center and coordinated by two diethyl\u00adnicotinamide (DENA) ligands, two 4-bromo\u00adbenzoate (PBB) anions and two water mol\u00adecules in a distorted octa\u00adhedral geometry. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 3.25\u2005(14)\u00b0. In the mol\u00adecule, the pyridine ring and the benzene ring are oriented at a dihedral angle of 77.24\u2005(5)\u00b0. In the crystal, inter\u00admolecular C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a two-dimensional network. Weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings of neighbouring mol\u00adecules [centroid\u2013centroid distance = 3.537\u2005(1)\u2005\u00c5] further consolidate the crystal packing. In the crystal structure of the title Mn N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C10H14N2O)2(H2O)2] = 0.026 wR(F 2) = 0.073 S = 1.09 4616 reflections233 parameters2 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.42 e \u00c5\u22123 \u0394\u03c1min = \u22120.39 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811031412/xu5285Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The six Ag atoms form a distorted octa\u00adhedron, with Ag\u22efAg distances in the range 2.933\u2005(1)\u20133.401\u2005(1)\u2005\u00c5. Each Ag atom is surrounded by one N atom and two thiol\u00adate S atoms from two deprotonated 2-hy\u00addroxy-1-naphthaldehyde thio\u00adsemi\u00adcarb\u00ada\u00adzone ligands. Each ligand coordinates three Ag atoms via a bridging thiol\u00adate S atom and a monodentate N atom, thus two Ag3S3 hexa\u00adgonal rings are linked together. Two dimethyl\u00adformamide solvent mol\u00adecules are located in four sets of sites with half-occupancy and form O\u22efH\u2014N hydrogen bonds to the complex mol\u00adecule. Intra\u00admolecular O\u2014H\u22efN hydrogen bonds are also present. The discrete hexa\u00adnuclear clusters are further linked through \u03c0\u2013\u03c0 inter\u00adactions into layers parallel to (001), the shortest distance between the centroids of aromatic rings being 3.698\u2005(2)\u2005\u00c5.In the title compound, [Ag DOI: Click here for additional data file.10.1107/S1600536812050155/yk2079Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The four nearest O atoms around each CuII ion form a distorted square-planar arrangement, the distorted square-pyramidal coordination being completed by the O atom of the water mol\u00adecule at a distance of 2.1525\u2005(16)\u2005\u00c5. The dihedral angle between the benzene ring and the carboxyl\u00adate group is 25.67\u2005(13)\u00b0 in one of the independent IB ligands and 6.44\u2005(11)\u00b0 in the other. The benzene rings of the two independent IB ligands are oriented at a dihedral angle of 86.61\u2005(7)\u00b0. In the crystal, O\u2014H\u22efO inter\u00adactions link the mol\u00adecules into a two-dimensional network. \u03c0\u2013\u03c0 contacts between the benzene rings [centroid\u2013centroid distances = 3.810\u2005(2) and 3.838\u2005(2)\u2005\u00c5] may further stabilize the structure.In the centrosymmetric binuclear title complex, [Cu N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 4(H2O)2] = 0.019wR(F2) = 0.045S = 1.163987 reflections207 parameters2 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.61 e \u00c5\u22123\u0394\u03c1min = \u22120.66 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812010367/bq2345Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "IIMnIII 2(C11H13NO4)2(CH3CO2)4(H2O)2]\u00b72CH3CN\u00b72H2O, there are two MnIII and one MnII atoms. The MnII atom lies on an inversion center and the MnIII\u2014MnII\u2014MnIII angle is therefore 180\u00b0, as required by crystallographic symmetry. The MnIII and MnII atoms are six-coordinated in a distorted octa\u00adhedral geometry. In the crystal, complex mol\u00adecules and solvent mol\u00adecules are linked into a three-dimensional network by O\u2014H\u22efO and O\u2014H\u22efN hydrogen-bonding inter\u00adactions. In the title complex, [Mn DOI: 10.1107/S1600536811027899/pv2425Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the complex, the MnII ion is six-coordinated in a considerably distorted octa\u00adhedral environment defined by four N atoms of the two chelating 2,2\u2032-bipyrimidine (bpym) ligands, one I\u2212 anion and one O atom of a water ligand. As a result of the different trans effects of the I and O atoms, the Mn\u2014N bond trans to the I atom is slightly longer than the Mn\u2014N bond trans to the O atom. The dihedral angle between the least-squares planes of the two bpym ligands [maximum deviation = 0.088\u2005(4)\u2005\u00c5] is 76.48\u2005(6)\u00b0. In the crystal, the complex cation, the anion and the solvent water mol\u00adecules are linked by inter\u00admolecular O\u2014H\u22efO, O\u2014H\u22efI and O\u2014H\u22efN hydrogen bonds.The asymmetric unit of the title compound, [MnI(C II complexes, see: Hong et al. 2(H2O)]I\u00b72H2O = 0.037 wR(F 2) = 0.096 S = 1.05 5707 reflections271 parametersH-atom parameters constrainedmax = 0.97 e \u00c5\u22123 \u0394\u03c1min = \u22121.15 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811037810/aa2026Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "As a result of the different trans effects of Br, N and O atoms, the Mn\u2014N bond trans to the Br atom is slightly longer than the Mn\u2014N bond trans to the N or O atoms. In the crystal, the dpa ligands are not planar, the dihedral angles between the two pyridine rings being 29.2\u2005(4) and 28.2\u2005(3)\u00b0. The complex cations and the Br\u2212 anions are linked by inter\u00admolecular O\u2014H\u22efBr and N\u2014H\u22efBr hydrogen bonds. Inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions are present between the pyridine rings, with a centroid\u2013centroid distance of 3.793\u2005(4)\u2005\u00c5.In the title compound, [MnBr(C II complexes with a di-2-pyridyl\u00adamine ligand, see: Bose et al. 2(H2O)]Br = 0.057 wR(F 2) = 0.174 S = 0.96 4215 reflections271 parametersH-atom parameters constrainedmax = 1.02 e \u00c5\u22123 \u0394\u03c1min = \u22121.02 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811048100/hy2487Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordination about each Rb atom is completed by a monodentate water mol\u00adecule and a phenolic O-atom donor which gives a bridging extension [Rb\u2014O range = 3.116\u2005(7)\u20133.135\u2005(5)\u2005\u00c5]. The polymeric structure is stabilized by inter\u00admolecular water O\u2014H\u22efOcarboxyl\u00adate hydrogen bonds and weak inter-ring \u03c0\u2013\u03c0 inter\u00adactions [minimum ring centroid separation = 3.620\u2005(4)\u2005\u00c5]. An intramolecular O\u2014H\u22efO hydrogen bond between phenol and carboxylate groups is also present.In the structure of title compound, [Rb(C DOI: 10.1107/S1600536811037561/nk2111Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "These chains are further extended into a three-dimensional network via O\u2014H\u22efO hydrogen-bonding inter\u00adactions and inter\u00adchain \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.662\u2005(2)\u2005\u00c5].In the title compound, [Co DOI: 10.1107/S1600536811019970/sj5153Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The central five-membered ring (Pro2) of the peptide mol\u00adecule has a disordered envelope conformation [occupancy ratio 0.879\u2005(2):0.121\u2005(2)] with the envelope flap atom, the central C atom of the three ring methylene groups, lying on alternate sides of the mean ring plane. The terminal five-membered ring (Pro4) also adopts an envelope conformation with the C atom of the methylene group closest to the carboxylic acid function as the envelope flap, and the six-membered tetra\u00adhydro\u00adpyrane ring shows a chair conformation. The tetra\u00adpeptide exists in a helical conformation, stabilized by an intra\u00admolecular hydrogen bond between the amide N\u2014H group of the heterocyclic \u03b1-amino acid Thp and the amide O atom of the 4-meth\u00adoxy\u00adbenzoyl group. This inter\u00adaction has a graph set motif of S(10) and serves to maintain a fairly rigid \u03b2-turn structure. In the crystal, the terminal hy\u00addroxy group forms a hydrogen bond with the amide O atom of Thp of a neighbouring mol\u00adecule, and the amide N\u2014H group at the opposite end of the mol\u00adecule forms a hydrogen bond with the amide O atom of Thp of another neighbouring mol\u00adecule. The combination of both inter\u00admolecular inter\u00adactions links the mol\u00adecules into an extended three-dimensional framework.The asymmetric unit of the title compound, C \u00c5b = 13.7414 (2) \u00c5c = 21.1929 (3) \u00c5V = 3162.48 (7) \u00c53Z = 4K\u03b1 radiationMo \u22121\u03bc = 0.16 mmT = 160 K0.28 \u00d7 0.20 \u00d7 0.18 mmNonius KappaCCD area-detector diffractometer53400 measured reflections9238 independent reflectionsI > 2\u03c3(I)7711 reflections with Rint = 0.044R[F2 > 2\u03c3(F2)] = 0.042wR(F2) = 0.103S = 1.029231 reflections433 parameters21 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.28 e \u00c5\u22123\u0394\u03c1min = \u22120.33 e \u00c5\u22123\u0394\u03c1Absolute structure: Flack & Bernardinelli 1999, 2000 \u25b6,Flack parameter: \u22120.02 (8)COLLECT I, global. DOI: Click here for additional data file.10.1107/S1600536813004546/nc2305Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536813004546/nc2305Isup3.cdxSupplementary material file. DOI: Click here for additional data file.10.1107/S1600536813004546/nc2305Isup4.cmlSupplementary material file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal structure, inter\u00admolecular O\u2014H\u22efO hydrogen bonds link the mol\u00adecules into centrosymmetric dimers, and \u03c0\u2013\u03c0 inter\u00adactions link these dimers into layers parallel to the ac plane. Weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions further consolidate the crystal packing.In the title mol\u00adecule, C DOI: 10.1107/S1600536810028394/cv2747Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The bis\u00ad(diphenyl\u00adphosphan\u00adyl)ethane ligand bridges an Ru\u2014Ru bond and the monodentate stibine ligand bonds to the third Ru atom. Both the stibine and phosphine ligands are equatorial with respect to the Ru3 triangle. Additionally, each Ru atom carries one equatorial and two axial terminal carbonyl ligands. The three stibine-substituted benzene rings make dihedral angles of 38.7\u2005(3), 71.5\u2005(3) and 70.0\u2005(3)\u00b0 with each other in mol\u00adecule A whereas these angles are 83.9\u2005(3), 88.2\u2005(3) and 56.8\u2005(3)\u00b0 in mol\u00adecule B. Similarly, the dihedral angles between the two benzene rings are 80.7\u2005(3) and 87.6\u2005(3)\u00b0 for the two diphenyl\u00adphosphanyl groups in mol\u00adecule A and 84.0\u2005(3) and 72.6\u2005(4)\u00b0 in mol\u00adecule B. In the crystal, mol\u00adecules are linked into tetra\u00admers via inter\u00admolecular C\u2014H\u22efO hydrogen bonds. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions further stabilize the crystal structure.The asymmetric unit of the title DOI: 10.1107/S1600536810054218/sj5087Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Sn\u2014Cl bond [2.366\u2005(2)\u2005\u00c5] trans to nitro\u00adgen is shorter than the others [2.438\u2005(2) and 2.414\u2005(2)\u2005\u00c5]. The N\u2014Sn\u2014N angle [76.19\u2005(11)\u00b0] is smaller than the O\u2014Sn\u2014N angle [87.89\u2005(10)\u00b0] in the Schiff base ligand. No classical inter\u00admolecular hydrogen-bonding inter\u00adactions are observed. The crystal packing exhibits \u03c0\u2013\u03c0 stacking inter\u00adactions, with a distance of 3.595\u2005(2)\u2005\u00c5 between the centroids of the phenolate ring and the benzene ring of the quinoline group of inversion-related mol\u00adecules.In the title compound, [Sn(C For a related structure, see: Takano & Shibahara 2008.20H19N2O3)Cl3] = 0.042wR(F2) = 0.101S = 1.045989 reflections292 parametersH-atom parameters constrainedmax = 1.51 e \u00c5\u22123\u0394\u03c1min = \u22121.24 e \u00c5\u22123\u0394\u03c1CrystalClear CrystalClear; data reduction: CrystalClear; program(s) used to solve structure: SIR2004 global, I. DOI: 10.1107/S1600536812002528/pk2385Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II title complex, [Ni(C7H4FO2)2(C6H6N2O)2(H2O)2], the NiII atom, located on an inversion center, is coordinated by two nicotinamide and two 4-fluoro\u00adbenzoate ligands and two water mol\u00adecules in a distorted N2O4 octa\u00adhedral geometry. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 8.95\u2005(8)\u00b0, while the pyridine ring and the benzene ring are oriented at a dihedral angle of 75.01\u2005(7)\u00b0. The water mol\u00adecule links the adjacent carboxyl\u00adate O atom via an intra\u00admolecular O\u2014H\u22efO hydrogen bond. In the crystal, O\u2014H\u22efO, N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds link the mol\u00adecules into a three-dimensional network. \u03c0\u2013\u03c0 stacking between parallel pyridine rings [centroid\u2013centroid distance = 3.7287\u2005(11)\u2005\u00c5] is also observed.In the mononuclear Ni N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.030 wR(F 2) = 0.077 S = 1.04 3220 reflections203 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.46 e \u00c5\u22123 \u0394\u03c1min = \u22120.57 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811044771/xu5359Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The AgI ion lies on a twofold rotation axis. The primary structure consists of a one-dimensional coordination polymer formed by the AgI ions and the bipyridyl azine ligand in which there is an inversion center at the mid-point of the N\u2014N bond. The nitrite anion inter\u00adacts with the AgI ion through a chelating \u03bc2 inter\u00adaction involving both O atoms. In the crystal, the coordination chains are parallel and inter\u00adact through Ag\u22ef\u03c0 [3.220\u2005(2)\u2005\u00c5] and \u03c0\u2013\u03c0 [3.489\u2005(3)\u2005\u00c5] inter\u00adactions.The asymmetric unit of the title compound, [Ag(NO For the al. 2005. For nit al. 2005; Blake e al. 1999; Cingola al. 1999; Fl\u00f6rke al. 1998; Tong et al. 2002. 2)(C14H14N4)] = 0.023 wR(F 2) = 0.048 S = 1.05 1693 reflections102 parametersH-atom parameters constrainedmax = 0.44 e \u00c5\u22123 \u0394\u03c1min = \u22120.44 e \u00c5\u22123 \u0394\u03c1 DENZO used to solve structure: SIR92 global, I. DOI: 10.1107/S1600536811028546/lh5278Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The PdII cations are bridged by the S atoms of two benzene\u00adthiol\u00adate ligands with different Pd\u2014S distances [2.2970\u2005(11) and 2.3676\u2005(11)\u2005\u00c5]. The coordination of the metal atom is completed by a chloride anion [2.3383\u2005(11)\u2005\u00c5] and a tri\u00adphenyl\u00adphosphane ligand [2.2787\u2005(11)\u2005\u00c5]. Weak C\u2014H\u22efCl inter\u00adactions are present between complex mol\u00adecules and the CHCl3 solvent mol\u00adecule. The latter is disordered over two positions in a 0.792\u2005(8):0.208\u2005(8) ratio. The crystal under investigation was found to be twinned by nonmerohedry, with a fraction of 73.4\u2005(1)% for the major twin component.The title compound, [Pd DOI: 10.1107/S1600536813019806/wm2753Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "V compound, [Nb2(C44H28N4)2O3], each Nb atom is seven-coordinated with three bridging O atoms and four N atoms from a chelating tetra\u00adphenyl\u00adporphyrinate anion. The Nb\u2014O bond lengths range from 1.757\u2005(6) to 2.331\u2005(6)\u2005\u00c5, and the average (niobium\u2013pyrrole N atom) distance is 2.239\u2005\u00c5. In the dinuclear mol\u00adecule, the Nb\u22efNb separation is 2.8200\u2005(8)\u2005\u00c5, and the dihedral angle between the two porphyrinate mean planes is 5.4\u2005(1)\u00b0. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions are present in the crystal structure.In the title dinuclear Nb DOI: 10.1107/S1600536811020538/xu5216Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The methoxide ligands form an asymmetric bridge between the two TiIV atoms [Ti\u2014O bond lengths of 1.9794\u2005(12) and 2.0603\u2005(12)\u2005\u00c5] with the two phenolato ligands occupying the remaining basal sites [Ti\u2014O 1.8218\u2005(11) and 1.8135\u2005(11)\u2005\u00c5]. The Ti\u2014O\u2014C phenolato bond angles are similar at 161.24\u2005(10) and 160.66\u2005(11)\u00b0. The methyl ligand attached to the metal atom has a Ti\u2014C bond length of 2.0878\u2005(17)\u2005\u00c5.The molecule of the title compound, [Ti DOI: 10.1107/S1600536813025634/gg2122Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The EuIII ion is eight-coordinated by two O atoms from two Hbdc ligands, three O atoms from two H2bdc ligands and three water mol\u00adecules, showing a distorted square-anti\u00adprismatic geometry. The EuIII ions are bridged by the carboxyl\u00adate groups of the Hbdc and H2bdc ligands, forming a chain along [110], with an Eu\u22efEu separation of 5.4594\u2005(3)\u2005\u00c5. These chains are further connected by inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, as well as \u03c0\u2013\u03c0 inter\u00adactions between the imidazole and benzene rings , into a three-dimensional supra\u00admolecular network.In the title one-dimensional coordination polymer, {[Eu(C H-benzimidazole-5,6-dicarboxyl\u00adate complexes, see: Fu et al. (C9H5N2O4)(H2O)3]\u00b72H2O = 0.024 wR(F 2) = 0.057 S = 1.04 3711 reflections328 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.63 e \u00c5\u22123 \u0394\u03c1min = \u22120.67 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, glogal. DOI: 10.1107/S1600536811033496/hy2459Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.838\u2005(1)\u2005\u00c5] and C\u2014H\u22efCl hydrogen bonds link adjacent mol\u00adecules into a chain structure along [101].In the title compound, [CuCl DOI: 10.1107/S160053681101333X/ng5145Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The apical PbII atom of each pyramid is 1.33059\u2005(3)\u2005\u00c5 above the basal S4 plane. The metal atom is surrounded by two chelating dithio\u00adphospho\u00adnate ligands binding through the S-donor atoms. The ligands are anisobidentate as the pyramid is comprised of Pb\u2014S bond lengths that vary substanti\u00adally , clearly indicating two short and two longer bond lengths. The P\u2014S bond lengths are also paired as shorter [1.9959\u2005(9) and 1.9877\u2005(8)\u2005\u00c5] and slightly longer [2.0115\u2005(9) and 2.0245\u2005(9)\u2005\u00c5], indicating an anisobidentate nature of the ligand whereby the shorter P\u2014S bond has more double-bond character than the other. The S\u2014Pb\u2014S (chelating) bond angles range from 71.841\u2005(18) to 72.692\u2005(19)\u00b0, whilst the Pb\u2014S\u2014P bond angles range from 84.70\u2005(3) to 90.51\u2005(3)\u00b0.The title compound, [Pb(C DOI: Click here for additional data file.10.1107/S1600536812046818/gk2529Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Fe atom is slightly closer to the substituted cyclo\u00adpenta\u00addienyl ring, with an Fe\u2013centroid distance of 1.6374\u2005(3)\u2005\u00c5 [1.6494\u2005(3)\u2005\u00c5 for the unsubstituted ring]. The amide group is essentially coplanar with the substituted cyclo\u00adpenta\u00addienyl ring, with an N\u2014C(O)\u2014C\u2014C torsion angle of 2.3\u2005(3)\u00b0.In the title compound, [Fe(C DOI: 10.1107/S1600536811026341/fj2441Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The four Fe\u2014N bond lengths range from 1.9142\u2005(12) to 1.9579\u2005(11)\u2005\u00c5, while the Fe\u2014P bonds are 2.2452\u2005(4) and 2.2506\u2005(4)\u2005\u00c5 [P\u2014Fe\u2014P = 165.523\u2005(14)\u00b0], consistent with FeII in a low-spin state. Unlike related Fe PNP complexes based on 2,6-diamino\u00adpyridine, the BF4 anions are not hydrogen bonded to the two NH groups of the pincer ligand but show instead anion\u2013\u03c0 inter\u00adactions with the triazine ring and acetonitrile mol\u00adecules in addition to ten C\u2014H\u22efF inter\u00adactions. Most remarkable among these is an anion\u2013\u03c0(triazine) inter\u00adaction with a short distance of 2.788\u2005(2)\u2005\u00c5 between one F and the centroid of the \u03c0-acidic triazine ring. The corresponding shortest distance between this F atom and a triazine carbon atom is 2.750\u2005(2)\u2005\u00c5. The two NH groups of the pincer ligand donate N\u2014H\u22efN hydrogen bonds to the triazine N atom of a neighbouring complex and to an uncoordinated acetonitrile mol\u00adecule. This last mol\u00adecule is in a side-on head-to-tail association with the second uncoordinated acetonitrile at C\u22efN distances of 3.467\u2005(2) and 3.569\u2005(2)\u2005\u00c5. In contrast to several related compounds with diamino\u00adpyridine- instead of diamino\u00adtriazine-based PNP ligands, the title crystal structure is remarkably well ordered. This suggests that the diamino\u00adtriazine moiety exerts notable crystal structure stabilizing effects.In the title compound, [Fe(CH DOI: 10.1107/S1600536811049804/gk2437Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The HfIV and Si atoms lie on the rotation axis with all other atoms being in general positions. The HfIV atom is six-coordinated by two N atoms from the N2,N2\u2032-(di\u00admethyl\u00adsilanedi\u00adyl)bis\u00ad(N-tert-butyl-3-methyl\u00adbenz\u00adimid\u00adamidate) ligand and four Cl\u2212 ions in a slightly distorted octa\u00adhedral geometry. The two amidinate moieties are connected through the central Si atom with Si\u2014N bond length of 1.762\u2005(3)\u2005\u00c5, generating the characteristic N\u2014C\u2014N\u2014Si\u2014N\u2014C\u2014N skeleton of a silyl-linked ansa-bis\u00ad(amidine) species.The symmetric title mol\u00adecule, [Hf(C Cl4] = 0.028wR(F2) = 0.061S = 1.022920 reflections169 parametersH-atom parameters constrainedmax = 0.53 e \u00c5\u22123\u0394\u03c1min = \u22120.29 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536813030328/sj5366Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the cation, the pyridinium rings attached to the central 1-aza\u00adniumyl-2-hy\u00addroxy\u00adethane fragment show an anti conformation, as indicated by the central C\u2014C\u2014C\u2014C torsion angle of \u2212178.1\u2005(4)\u00b0, and they are inclined to one another by 25.7\u2005(2)\u00b0. In the crystal, the cations and anions are linked through N\u2014H\u22efCl and O\u2014H\u22efCl hydrogen bonds. There are also \u03c0\u2013\u03c0 contacts [centroid\u2013centroid distance = 3.788\u2005(3)\u2005\u00c5] and a number of C\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions are present, consolidating the formation of a three-dimensional structure.The asymmetric unit of the title compound, (C DOI: Click here for additional data file.10.1107/S1600536812050817/su2540Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atom in the title compound, [Zn(C5H10NS2)2(C12H8N2)], exists in a distorted cis-octa\u00adhedral N2S4 donor set defined by two chelating dithio\u00adcarbamate anions as well as a 1,10-phenanthroline ligand. Each of the ligands coordinates in a symmetric mode. The crystal packing is stabilized by weak C\u2014H\u22efS, C\u2014H\u22ef\u03c0(ZnS2C) and \u03c0\u2013\u03c0 [ring centroid distance between centrosymmetrically related pyridyl rings = 3.5955\u2005(13)\u2005\u00c5] inter\u00adactions.The Zn DOI: 10.1107/S1600536811012499/hb5835Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "A zigzag chain structure is formed, extending along (001). Each chain is surrounded by three others which are inter\u00adconnected through weak C=O\u22ef\u03c0pyrid\u00adyl [O\u22efcentroid = 2.999\u2005(3)\u2005\u00c5] and \u03c0pyrid\u00adyl\u2013\u03c0pyrid\u00adyl inter\u00adactions [minimum ring centroid separation = 4.014\u2005(2)\u2005\u00c5], giving a three-dimensional framework. In the title polymer, [ZnCl DOI: 10.1107/S160053681104671X/zs2159Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ZnII atom is coordinated by three O atoms from two 1-naphthyl\u00adacetate ligands, one monodentate and the other asymmetric bidentate chelate, and two N atoms from a 5,5\u2032-dimethyl-2,2\u2032-bipyridine ligand, giving an irregular environment. In the crystal, the complex mol\u00adecules are inter\u00adlinked through the water mol\u00adecule by O\u2014H\u22efOcarboxyl\u00adate hydrogen bonds, together with weak C\u2014H\u22efO and bipyridine ring \u03c0\u2013\u03c0 stacking inter\u00adactions [ring centroid separation = 3.761\u2005(2)\u2005\u00c5], giving a two-dimensional network structure.In the title compound, [Zn(C DOI: 10.1107/S1600536811013353/zs2105Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the cluster, the CaII atom is connected to two S atoms of an Fe2S2 core [Ca\u2014S = 2.7463\u2005(8) and 2.7523\u2005(8)\u2005\u00c5]. No Fe\u2014Ca bonds are formed [Fe\u22efCa = 3.6708\u2005(6) and 3.5802\u2005(6)\u2005\u00c5]. There are five close C\u2013H\u22efO\u2013C contacts in the crystal structure.Reaction between [Fe DOI: Click here for additional data file.10.1107/S1600536812048039/zl2517Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Z\u2032 = 2) in the asymmmetric unit of the title compound, [CuBr(C16H32N4)]Br\u00b72H2O. The title crystal consists of two [Cu(C16H32N4)]2+ cations, two Br\u2212 anions and four uncoordinated water mol\u00adecules. The metal atom is five-coordinate square pyramidal, with a long apical Cu\u2014Br bond [2.9734\u2005(11) and 2.9229\u2005(11)\u2005\u00c5 in the two cations]. The two cations form a loosely associated dimer through the formation of hydrogen bonds between both N\u2014H and O\u2014H and Br\u2212. In addition, there is a network of N\u2014H\u22efBr, O\u2014H\u22efBr and N\u2014H\u22efO hydrogen bonds, leading to the formation of a chain structure.There are two formula units ( DOI: 10.1107/S1600536811012852/bv2173Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ligands form a slightly distorted square-planar coordination environment for the Rh(I) atom. An intra\u00admolecular P\u2013Rh\u2013P bite angle of 83.91\u2005(2)\u00b0 is observed. The dihedral angle between the P\u2014Rh\u2014P and the X\u2014Rh\u2014X planes (X is the centroid of a double bond) is 14.0\u2005(1)\u00b0. The BF4 anion is disordered over two positions in a 0.515\u2005(7):0.485\u2005(7) ratio.The title compound, [Rh(C DOI: 10.1107/S1600536810039577/si2281Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The monodentate-coordinated carboxyl\u00adate group is involved in an intra\u00admolecular O\u2014H\u22efO hydrogen bond with the coordinated water mol\u00adecule. In the crystal structure, inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into layers parallel to the ab plane. The crystal packing is further stabilized by weak C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions indicated by the short distance of 3.6181\u2005(8)\u2005\u00c5 between the centroids of the benzene and pyridine rings of neighbouring mol\u00adecules.In the title complex, [Co(C N,N-diethyl\u00adnicotinamide, see: Krishnamachari 2(C6H6N2O)(H2O)2] = 0.022 wR(F 2) = 0.050 S = 1.01 4838 reflections333 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.29 e \u00c5\u22123 \u0394\u03c1min = \u22120.24 e \u00c5\u22123 \u0394\u03c1Absolute structure: Flack 1983, 1761 FrFlack parameter: 0.015 (7) APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S160053681002194X/cv2727sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S160053681002194X/cv2727Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the cation, the CuII atom shows a slightly distorted square-pyramidal coordination geometry provided by a pair of \u03bc-OH\u2212 anions and by the N atoms of a chelate tmen ligand in the basal plane. The apical position is statistically occupied by the O atom of a half-occupancy water mol\u00adecule. The F atoms of the anion are disordered over three sets of sites with occupancies of 0.598\u2005(9):0.269\u2005(6):0.134\u2005(8). The crystal packing is governed by ionic forces as well as by O\u2014H\u22efF hydrogen bonds.The title compound, [Cu For add al. 1984.2(OH)2(C6H16N2)2(H2O)](BF4)2 = 0.046wR(F2) = 0.143S = 1.122118 reflections168 parameters1 restraintH-atom parameters constrainedmax = 1.01 e \u00c5\u22123\u0394\u03c1min = \u22120.61 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 I, global, New_Global_Publ_Block. DOI: 10.1107/S1600536812021836/rz2749Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Cl\u2014Cu\u2014N angles of 90.55\u2005(9) and 89.45\u2005(9)\u00b0 are close to ideal values. In the crystal, weak \u03c0\u2013\u03c0 stacking inter\u00adactions are observed between inversion-related benzene rings [centroid\u2013centroid distance = 4.0028\u2005(6)\u2005\u00c5].In the title compound, [CuCl DOI: 10.1107/S1600536812015486/bh2425Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Eucranium Brull\u00e9\u00e9 has been revised and now includes six species: E. arachnoides Brull\u00e9\u00e9, E. belenae Ocampo new species, E. cyclosoma Burmeister, E. dentifrons Gu\u00e9\u00e9rin-M\u00e9\u00e9neville, E. planicolle Burmeister, and E. simplicifrons Fairmaire. Eucranium pulvinatum Burmeister is a new junior synonym of Eucranium arachnoides Brull\u00e9\u00e9, and Eucranium lepidum Burmeister is a new junior synonym of E. dentifrons Gu\u00e9\u00e9rin-M\u00e9\u00e9neville. The following lectotypes and neotypes are designated: Eucranium pulvinatum Burmeister, lectotype; Eucranium planicolle Burmeister, lectotype; Psammotrupes dentifrons Gu\u00e9\u00e9rin-M\u00e9\u00e9neville, neotype; and Eucranium lepidum Burmeister, neotype. Description of the genus and new species, diagnosis and illustrations, and distribution maps are provided for all species. A key to the species of this genus is provided, and the biology and conservation status of the species are discussed.The South American genus Anomiopsoides Blackwelder, Ennearabdus van Lansberge, Eucranium Brull\u00e9\u00e9, and Glyphoderus Westwood (Eucranium consists of six species. The genus is endemic to Argentina and distributed in the Monte and Chacoan biogeographic provinces (based on The tribe Eucraniini (Scarabaeidae: Scarabaeinae) constitutes a monophyletic group and includes four genera, Westwood . This wobased on schema; based on .Eucranium was originally described by Brull\u00e9\u00e9 (1834) for one species, E. arachnoides Brull\u00e9\u00e9 (1834). The name was originally proposed by Dejean . Gu\u00e9\u00e9rin-M\u00e9\u00e9neville . Laporte (1840) described Pachysoma (Cyclodema) lacordairei (= E. arachnoides) and indicated that this was the only species of Pachysoma in America. Blanchard (Anomiopsis aelianus (= E. arachnoides). In 1845 Blanchard redescribed A. aelianus and referred to E. arachnoides and A. dioscorides with indication of their similarity to A. aelianus. Also Blanchard . Lacordaire (Eucranium to Brull\u00e9\u00e9 (1834) and provided a synonymy list . Burmeister . Mouth parts and male genitalia were dissected and cleaned in a dilute solution (\u223c?10%) of potassium hydroxide and neutralized in a dilute solution (\u223c? 10%) of acetic acid. The male genitalia were placed in a glycerin-filled vial pinned under the specimen.Body measurements, puncture density, puncture size, and density of setae were based on the following standards: Body length was measured from the middle of the anterior margin of the pronotum (at the middle) to the apex of the elytra, plus head length from the apex of clypeal process to the base of the head . Body width was measured across mid-pronotum. Puncture density was considered \u201c\u201cdense\u201d\u201d if punctures were nearly confluent to less than 2 puncture diameters apart, \u201c\u201cmoderately dense\u201d\u201d if punctures were 2\u20136 diameters apart, and \u201c\u201csparse\u201d\u201d if punctures were separated by more than 6 diameters. Puncture size was defined as \u201c\u201csmall\u201d\u201d if punctures were 0.02 mm or smaller, \u201c\u201cmoderate\u201d\u201d if 0.02\u20130.07 mm, and \u201c\u201clarge\u201d\u201d if 0.07 mm or larger. Setae were defined as \u201c\u201csparse\u201d\u201d if there were few setae, \u201c\u201cmoderately dense\u201d\u201d if the surface was visible but with many setae, and \u201c\u201cdense\u201d\u201d if the surface was not visible through the setae. Elytral carinae were counted from the elytral suture. Specimen labels were copied literally using \u201c\u201c/\u201d\u201d between lines and \u201c\u201c;\u201d\u201d between labels.Neotypes and Lectotypes were designated to provide the nomenclatural stability of the taxon studied, according to the Article 72 of the International Code of Zoological Nomenclature (1999).CMNC: Canadian Museum of Nature, Ottawa, Canada .HECO: Hope Entomological Museum, Oxford, England (Mann D).IAZA: Instituto Argentino de Investigaciones de las Zonas \u00c1\u00c1ridas, Mendoza, Argentina (F Ocampo).IMLA: Fundaci\u00f3\u00f3n e Instituto Miguel Lillo, Universidad Nacional de Tucum\u00e1\u00e1n, Tucum\u00e1\u00e1n, Argentina (MV Colomo).LEMQ: Lyman Entomological Museum, Mc Gill University, Quebec, Canada (S Boucher).MACN: Museo Argentino de Ciencias Naturales, Buenos Aires, Argentina (A Roig).MNHN: Mus\u00e9\u00e9um National d'Histoire Naturelle, Paris, France (O Montreuil).MLPA: Museo de La Plata, La Plata, Argentina (A Lanteri).UNSM: University of Nebraska State Museum, Lincoln, NE, USA (BC Ratcliffe).USNM: United States National Museum, Washington D.C. USA (D Furth).Specimens for this research were collected or borrowed from and deposited in the following institutions and collections:Eucranium genus were described and recognized based mostly on the shape and length of the clypeal processes, and the pronotal and elytral sculptures. These characters rendered variable within species and were not reliable for species identification or description. In this work, new characters were explored and used to define species. Among these are Elytral pseudoepipleuron, pseudoepipleural angle with respect to elytral disc, elytral 8th striae (shape and sculpture), and shape and development of mesotibial spurs. Species male genitalia were studied in order to find species-specific patterns in the shape of the paremares, but the findings were not informative at this level. Internal sacs of paremeres were extracted and studied and these structures provided highly valuable information for phylogenetic analysis, however, they are impractical for species identification. Nevertheless, the information from the paremeres internal sacs is currently being used in a separate project on Eucraniini evolutionary biology .Traditionally species in the Eucranium Brull\u00e9\u00e9 1834: 286.EucraniumNomen nudum).EucraniumAnomiopsisnecA. dioscoridesAnomiopsis Westwood 1838: 159, PsammotrupesP. dentifronsPsammotrupesCyclodema Laporte 1840: 68, (as subgenus of Pachysoma Mac Leay), junior synonym. Type species Pachysoma lacordairei Laporte 1840: 68.Anomiopsis Westwood 1838; Eucranium).Eucranium Brull\u00e9\u00e9; Eucranium Brull\u00e9\u00e9; Eucranium Brull\u00e9\u00e9; Eucranium Brull\u00e9\u00e9; Eucranium Brull\u00e9\u00e9; Eucranium Brull\u00e9\u00e9; Eucranium Brull\u00e9\u00e9; Eucranium arachnoides Brull\u00e9\u00e9 1834, by monotypy.Type species: DiagnosisThe Eucraniini genus thacan be distinguished from other members of the New World Scarabaeinae by the following combination of characters: Body relatively large (13\u201330 mm), black , 22, 24;RedescriptionHead . Venter: Surface smooth, glabrous or sparsely setose, prosternum pentagonal, anterior margins slightly concave. Mesosternum wider than long, mesometasternum suture visible or not. Metasternum flat, strongly narrowed in middle (metacoxae contiguous). Metepisternum 2.5\u20133 times longer than wide (at base). Ventrites narrower at middle. Pygidium with base grooved medially; disc slightly convex, sparsely punctate, punctures variable. Legs . These differences are not consistent among individuals of the same population. Based on the species concept used in this work to recognize Eucranium species, all these differences are attributed to intraspecific variation. Molecular information is needed to elucidate weather isolated populations ; Argerich (1); Bah\u00ed\u00eda Blanca (4); Bah\u00ed\u00eda San Blas (1); Bajo Hondo (4); Carmen de Patagones (2); Estancia Barrau (6); Felipe Sol\u00e1\u00e1 (7); La Colina (1); Maza (3); Villa Iris (1). C\u00f3\u00f3rdoba: no data (6); \u201c\u201cSur de C\u00f3\u00f3rdoba\u201d\u201d (1); San Javier (2); Las Rosas (3); Potrero de G\u00f3\u00f3mez (1); Yacanto de San Javier (1). La Pampa: no more data (1); Gaviotas (1); Santa Rosa (1); Victorica (4). Mendoza: no more data (1); Agua Escondida (2); Aguada de los Ciegos (1); Arroyo el Rosario, Puesto las Gateadas (1); Arroyo La Rinconada (1); Base del Volc\u00e1\u00e1n Diamante (1); Blanco Encalada (1); Caverna de los Tigres (1); Confluencia r\u00ed\u00edo Diamante and r\u00ed\u00edo Salado (1); Costa de Araujo (1); Dique Agua del Toro (4); Dique Agua del Toro (20 km S) (3); Dique El Carrizal (3); Divisadero (2); El Mollar (2); El Nihuil, M\u00e9\u00e9danos (1); Embalse El Nihuil (4); Fort\u00ed\u00edn Malarg\u00fc\u00fce (1); Huayquer\u00ed\u00edas (1); From RN 40 to Puesto Alvarado (2); Malarg\u00fc\u00fce (no more data) (4); Malarg\u00fc\u00fce, Los Corrales (5); Monte Com\u00e1\u00e1n (1); \u00d1\u00d1acu\u00f1\u00f1\u00e1\u00e1n (9); Pareditas (2); Reserva de la Bi\u00f3\u00f3sfera \u00d1\u00d1acu\u00f1\u00f1\u00e1\u00e1n (11); Reserva Natural Laguna del Diamante (15 km SE) (19); Reserva Natural La Payunia, Puesto La Senillosa (1); Reserva Natural La Payunia, Los Relinchos (11); Reserva Natural La Payunia, Valle del Saino (1); RN 40 and Arroyo Yaucha (3); RN 40 (km 143) (1); RN 40 (S of Pareditas) (2); Road to Paso de Los Tigres (1); Salar del Nihuil (2); San Rafael (10). R\u00ed\u00edo Negro: Coronel G\u00f3\u00f3mez (2); R\u00ed\u00edo Colorado (3). San Luis: no more data (1); Arizona (19); Balde (2); San Luis, Departamento Capital (7); El Volc\u00e1\u00e1n (1); San Ger\u00f3\u00f3nimo (1). \u201c\u201cPatagonia\u201d\u201d no more data (1).ARGENTINA . Buenos Temporal distributionJanuary (79); February (12); March (12); April (5); May (1); July (2); August (4); September (7); October (4); November (15); December (51); no data (41).Biology and conservationEucranium species. Populations of this species generally have a small, patchy distribution and consequently susceptibly to local extinction if changes in the environmental conditions occur. The only known populations of E. arachnoides that are currently in a protected area are those from Reserva Natural \u00d1\u00d1acu\u00f1\u00f1\u00e1\u00e1n and Reserva Natural La Payunia in the Mendoza province.Biology and behavior of this species were recently discussed by Zunino et al. , MontereType materialEucranium belenae Ocampo holotype male at IAZA labeled: \u201c\u201cARGENTINA: Mendoza / R.N 142 km 107. N. Rva. / Telteca / 510m. 32\u00b0\u00b015\u2032?12\u2033?S / 67\u00b0\u00b049\u2032?13\u2033?W. 30/III/2009/ F. C. Ocampo\u201d\u201d; \u201c\u201cEucranium / belenae / HOLOTYPE / F. C. Ocampo\u201d\u201d, Allotype female labeled as holotype except: \u201c\u201cEucranium / belenae / ALLOTYPE / F. C. Ocampo.\u201d\u201d Twenty eight male and twenty female paratypes at IAZA: labeled as holotype. Eleven male and six female paratypes at IAZA labeled: \u201c\u201cARGENTINA: Mendoza / Reserva Telteca. 32\u00b0\u00b022\u2248?59.58\u2033? S, / 68\u00b0\u00b003\u2032?14.16\u2033? W. 548m. / 11-IV-2008. Col. L. Mu\u00f1\u00f1oz.\u201d\u201d Fourteen male and nine female paratypes at IAZA labeled as previous except: \u201c\u201c12-IV-2008\u201d\u201d. Three male paratypes at IAZA labeled as previous except: \u201c\u201c13-IV-2008\u201d\u201d. Three male paratypes at IAZA labeled as previous except: \u201c\u201c14-IV-2008\u201d\u201d. One male paratype at IAZA labeled as previous except: \u201c\u201c15-IV-2008\u201d\u201d. Three male and two female paratypes at IAZA labeled: \u201c\u201cARGENTINA: Mendoza / R.N. 142 Km 107, N Rva. / Telteca. 510m. 32\u00b0\u00b015\u2032?12\u2033?S / 67\u00b0\u00b049\u2032?13\u2033?W. 1-III-2009. / KS Sheldon, FC Ocampo.\u201d\u201d Four male and eight female paratype at IAZA labeled as previous except: \u201c\u201c1/III/2009\u201d\u201d. Four male and Four female paratype at IAZA labeled as previous except: \u201c\u201cFC Ocampo, K Sheldon\u201d\u201d and \u201c\u201c17/III/2009\u201d\u201d. Seven male and three female paratypes at IAZA labeled: \u201c\u201cARGENTINA: Mendoza / Lavalle. Telteca. 32\u00b0\u00b022\u2032?59.58\u2033?S. / 68\u00b0\u00b003\u2032?14.16\u2033?W. 548m. 05-II-2008. / Col. F. Ocampo, E. Ruiz, G. San Blas.\u201d\u201d One paratype at IAZA labeled: \u201c\u201cARGENTINA: Mendoza / Lavalle. Telteca. 32\u00b0\u00b022\u2032?59.58\u2033?S. / 68\u00b0\u00b003\u2032?14.16\u2033?W. 548m. 27-XI-2007. / Col. F. Ocampo\u201d\u201d. One male and one female paratypes at IAZA labeled: \u201c\u201cARGENTINA: Lavalle / Puente R\u00ed\u00edo Mendoza. 21 Feb. 2006. E. Ruiz.\u201d\u201d One male and one female paratypes at IAZA labeled: \u201c\u201cARGENTINA: Mendoza / Reserva Telteca. 563m. S32\u00b0\u00b023\u2032?33\u2033? W68\u00b0\u00b003\u2032?00\u2033? / Jan-3-2002. F. C. Ocampo.\u201d\u201d Four female paratypes at IAZA labeled: \u201c\u201cR.A. Mza. Lavalle / Telteca / 3/2-14/3 /1995 / S. Roig / G. Flores.\u201d\u201d Four female paratypes at IAZA labeled: \u201c\u201cRA. Mza. Lavalle / Telteca /2/11 \u2013 1/12 /1994 / G. Flores.\u201d\u201d One male and two female paratypes at IAZA labeled: \u201c\u201cRa. Mendoza Tel- / teca 17/VIII-24/IX / 1996. Flores/Roig.\u201d\u201d One male and one female paratypes at IAZA labeled: \u201c\u201cRa. Mendoza Tel- / teca 25/XI-25/XII / 1995. Flores/Roig.\u201d\u201d One male and one female paratypes at IAZA labeled: \u201c\u201cRa. Mendoza Tel- / teca 25/IX-5/XI / 1996. Flores/Roig\u201d\u201d. One male paratype at IAZA labeled: \u201c\u201cRa. Mendoza Tel- / teca 14/8 -24/9/ / 1995 Flores/Roig.\u201d\u201d Two female paratype at IAZA labeled: \u201c\u201cMendoza, Lavalle / Telteca 15-2 al 25-3-96. Col. G. Flores / IADIZA.\u201d\u201d One female paratype at IAZA labeled as previous except: \u201c\u201c15-4-95\u201d\u201d. One male and three female paratypes at IAZA labeled: \u201c\u201cRa. Mendoza Tel- / teca 3/II -14/III / 1995. Flores/Roig.\u201d\u201d One male and two female paratype at IAZA labeled: Mendoza, Lavalle / Telteca 1 al 15/12/ 94 Flores/Roig / IADIZA.\u201d\u201d One female paratype at IAZA labeled: \u201c\u201cMendoza, Lavalle / Telteca 10/10 al 3/12/96 / Col. Gonzalez / IADIZA.\u201d\u201d Three male paratypes at IAZA labeled: \u201c\u201cRA. Mza. Lavalle / Telteca / 01.III.94 / G. Flores / IADIZA.\u201d\u201d One male paratype at IAZA labeled: \u201c\u201cMendoza. Lavalle / Parque Telteca / 10.5.93 / M. Gonz\u00e1\u00e1lez / IADIZA\u201d\u201d One female paratype at IAZA labeled: \u201c\u201cMendoza Lavalle Telteca 3-XII-96 / 6-I-97 Flores-Roig.\u201d\u201d One female paratype at IAZA labeled: \u201c\u201cMendoza Lavalle Telteca 25/9-31/10 / 1995 Flores/Roig.\u201d\u201d One female paratype at IAZA labeled: \u201c\u201cMendoza Lavalle Telteca 2/V-14/6 / 96 Flores/Roig / IADIZA.\u201d\u201d One male paratype at IAZA labeled: \u201c\u201cRA. Mza. Lavalle / El Enc\u00f3\u00f3n 12-IV-84 / IADIZA\u201d\u201d; \u201c\u201cCE.000131 / IADIZA\u201d\u201d. One male paratype at IAZA labeled: \u201c\u201cRA.Mza. Lavalle / Rva. Telteca / 15/X/03 / col. G. Debandi.\u201d\u201d One female paratype at IAZA labeled: \u201c\u201cRA. Mza. Lavalle / Telteca 01.III.94. / G. Flores / IADIZA.\u201d\u201d Three male and one female paratypes at IAZA with no data. All paratypes with a yellow paratype label: \u201c\u201cEucranium / belenae / PARATYPE / F. C. Ocampo.\u201d\u201dType localityArgentina, Mendoza, RN 142, km 107, 32\u00b0\u00b0 15\u2032? 12\u2033? S - 67\u00b0\u00b0 49\u2032? 13\u2033? W.DiagnosisE. belenae can be distinguished from other Eucranium species by the following combination of characters: Elytron with pseudoepipleuron not developed; elytral disc with interstriae becoming slightly convex toward margin, 8th stria slightly sulcate; apex of mesotarsus reaching apex of outer mesotibial spur or not ; outer mal axis) ; RN 142 Km 107 (114); RN 142 and R\u00ed\u00edo Mendoza (2); Reserva Natural Telteca (27); Telteca (94).ARGENTINA . MendozaTemporal distributionJan (11); Feb (27); March (59); April (66); May (2); June (1); August (4); September (7); October (2); November (14); December (15).Biology and conservationE. belenae were observed carrying goat pellets and small pieces of dry horse dung at daylight hours ; Andalgal\u00e1\u00e1 (2); Barranca Larga (4); Bel\u00e9\u00e9n (3); Capillitas (3); Corral Quemado (3); El Arenal (1); RP 47 N of Capillitas (9); El Ingenio (1); Hualf\u00ed\u00edn (3); Isla de Sauce (1); Loma Negra (2); Pipanaco (1); Punta de Balasto, 12 Km W. Campo El Arenal (3); Punta de Balasto (S. of Santa Maria) (28); RN 40 KM 892 (3). La Rioja: no more data (4); Aimogasta (1); Aminga (1); Anillaco (6); Anillaco (2 km N) (12); RN 40 E of Guandacol (1); La Rioja (1). Salta: La Caldera, Campo Alegre (1). Tucum\u00e1\u00e1n: Taf\u00ed\u00ed del Valle (1).ARGENTINA . CatamarTemporal distributionJanuary (21); February (52); March (7); November (23); December (4).Biology and conservationE. cyclosoma were observed carrying small pieces of dry horse dung at daylight hours over sand dunes in Catamarca province (Specimens of province . ConservEucranium dentifrons (Psammotrupes)ntifrons : 46 and so these species are placed in synonymy.Based on morphological evidence it was concluded that there are no differences between Eucranium dentifrons presents considerable variation in pronotal and elytral sculpture. Variation in puncture size and density on the pronotum and elytra is found among specimens of the same population and among populations. Variation is also found in rugosity on elytral interval seven, been in most specimens obvious and in some specimens slightly evident . These differences are more obvious among specimens from Neuqu\u00e9\u00e9n and western R\u00ed\u00edio Negro province.DistributionChubut: no data (1); Gaiman (1); Puerto Madryn (CENPAT) (5); Telsen (2); Pen\u00ed\u00ednsula Valdez, Estancia la Irma (5); Pen\u00ed\u00ednsula Valdez, Estancia Los M\u00e9\u00e9danos (5); Pen\u00ed\u00ednsula Valdez, Estancia San Pablo (23); Pen\u00ed\u00ednsula Valdez (no more data) (8); Pen\u00ed\u00ednsula Valdez, Playa Fracaso (1); Pen\u00ed\u00ednsula Valdez, Puerto Pir\u00e1\u00e1mides (1); Pen\u00ed\u00ednsula Valdez, Punta Delgada (5). Neuqu\u00e9\u00e9n: Aguada Florencia (2); Arroyo Pic\u00fa\u00fan Leuf\u00fa\u00fa (1); Colonia Centenario (1); Las Lajas (19 Km S) (4); Las Lajas, Cerro de la Cuchilla (4); Neuqu\u00e9\u00e9n (1); Pic\u00fa\u00fan Leuf\u00fa\u00fa (11); Piedra del \u00c1\u00c1guila (6); Plaza Huincul (16); R\u00ed\u00edo Agrio (7); RN 40 Km 2396, S of Las Lajas (14); RN 40, Bajada del Agrio (1); RN 40, El Marucho (1); Villa El Choc\u00f3\u00f3n(2 km W) (3); Zapala (4). R\u00ed\u00edo Negro: Barrancas del Gualicho (1); Cipolletti (1); Coronel Juan Jos\u00e9\u00e9 G\u00f3\u00f3mez (2); General Roca (1); San Antonio Oeste, Las Grutas (8).ARGENTINA . Chubut:Temporal distributionJanuary (38); February (62); March (16); April (8); September (1); October (6); November (2); December (12).Biology and conservationE. dentifrons have been observed caring and provisioning their borrows with guanaco dung pellets and small pieces of dry horse dung at daylight over sand dunes in Pen\u00ed\u00ednsula Valdez, Chubut, and in Choc\u00f3\u00f3n and near Las Lajas, Neuqu\u00e9\u00e9n (Specimens of rvation) .E. dentifrons is known to occur is Pen\u00ed\u00ednsula Vald\u00e9\u00e9z in Chubut province.Conservation status of this species has not been assessed. The only protected area where Eucranium planicolleType materialEucranium planicolle Burmeister lectotype male at MACN labeled: \u201c\u201cPampa / occid.\u201d\u201d; \u201c\u201cEucranium / planicolle / planicolle 6\u201d\u201d; \u201c\u201cEucranium / planicolle / Burm. / LECTOTYPE / F. C. Ocampo det. 2009\u201d\u201d. Lectotype here designated. One paralectotype male at MACN labeled as lactotype except: \u201c\u201cplanicolle 5\u201d\u201d.DiagnosisE. planicolle can be distinguished from other Eucranium species by the following combination of characters: Elytron with well defined pseudoepipleuron, pseudoepipleuron forming an \u223c?45\u201360\u00b0\u00b0 angle with elytral disc ; Bah\u00ed\u00eda Blanca (3); Bah\u00ed\u00eda San Blas (1); Bajo Hondo (6); Coronel Pringles (1); Estaci\u00f3\u00f3n Delta near Monte Hermoso (13); Estancia Barrau (30 Km SW Villa Iris) (8); Monte Hermoso (3); Villa Iris (7). La Pampa: no more data (1); Anguil (1). Mendoza: no more data (6); 25 de Mayo (1); Agua Escondida (1); Dique Agua del Toro (2); Dique Agua del Toro (20 Km S.) (2); Monte Com\u00e1\u00e1n (1); RP 143, Km 33 (1); Pareditas (3 Km S) (1); Pareditas (10 Km S) (1); Pareditas (22 Km S.) (1); Piedra P\u00f3\u00f3mez (1); Reserva Natural Laguna del Diamante, 10 Km E. (1); RN 40, Puesto Alvarado (1); RN 40 and Arroyo Yaucha (1); RN 40, S of Pareditas (4); RP 150 (1). R\u00ed\u00edo Negro: no more data (2); R\u00ed\u00edo Colorado (8); RP 4 (2). San Luis: Departamento Capital (5).ARGENTINA . No dataTemporal distributionJanuary (3); February (15); March (2); April (1); October (2); November (16); December (31).Biology and conservationE. planicolle are known to be diurnal and have been observed caring and provisioning their borrows with goat dung pellets in Mendoza province . Conservation status of this species has not been assessed.Specimens of Eucranium simplicifrons Fairmaire 1873: 608.DiagnosisE. simplicifrons can be distinguished from other Eucranium species by the following combination of characters: Elytron with or without pseudoepipleuron, if present, pseudoepipleuron forming an <65\u00b0\u00b0 angle with elytral disc ; Choya (8); El Charco (4); Fern\u00e1\u00e1ndez (1); Guasay\u00e1\u00e1n (1); Ram\u00ed\u00edrez deVelezco (1 Km N) (2).ARGENTINA . SantiagTemporal distributionFebruary (1); April (1); August (4); October (9); November (2).Biology and conservationE. simplicifrons. Conservation status of this species has not been assessed; the species does not occur in any protected area.With the exception that the species is diurnal nothing is known about the biology of th striae carinated, carina sharp and reflexed or rounded and reflexed1. Elytron with well defined pseudoepipleuron, pseudoepipleura forming a 45\u201360\u00b0\u00b0 angle with elytral disc , 23; ely2th stria not carinated, or if carinated, carinae poorly defined and never reflexed1\u2032?. Elytron with or without pseudoepipleuron, if present pseudoepipleuron forming a <65\u00b0\u00b0 angle with elytral disc , 15, 25;3th stria carinated, carina sharp 2. Elytron with outer margin of 8na sharp , 23; elyEucranium planicolle Burmeister (neville) 3. Elytron with pseudoepipleuron absent43\u2032?. Elytron with pseudoepipleuron present, sometimes poorly developedEucranium arachnoides Brull\u00e9\u00e9 ; outer mesotibial spur distinctively broad at apical 1/2, obviously asymmetrical ; mesotibial outer spur slightly broad on apical 1/3, nearly symmetrical . SantiagEucranium simplicifrons Lacordaireth stria slightly sulcate; west central Argentina (northeastern Mendoza province)5. Elytral disc with interstriae becoming slightly convex toward apical margin, 8Eucranium belenae Ocampo sp. nov. 5\u2032?. Elytral disc with interstriae smooth, evenly flat, 8Eucranium cyclosoma Burmeister . Thus, the Monte and Chacoan biota have multiple origins with most genera being from Neotropical origin followed by groups with Patagonian or Andean affinities.Morphological divergence of n biotas . South An biotas . In the Eucranium are distributed across a \u223c?2000 km long (North-South) and 500 km wide (East West) range. Nevertheless, species in this genus show little sympatry, E. arachnoides and E. planicolle partially share they distributional range, while the rest of the species, E. belenae, E. cyclosoma, E. dentifrons, and E. simplicifrons are isolated from other species in the genus or only share a few localities (ex. E. arachnoides and E. dentifrons in R\u00ed\u00edo Negro province). Eucranium species have high endemicity and populations have patchy distributions that make them susceptibly to local extinction if changes in the environmental conditions occur. Nothing is known for Eucranium species' population dynamics or habitat conservation status. Only two species, E. belenae and E. dentifrons are distributed within natural reserves or protected areas. It is well documented that there are genetic implications for small population size, among these it is a decline of genomic variation resulting from allelic loss (and Carrol (1997) for the long term viability of a population it is important for it to maintain genetic variability which would enable the population to adaptively tolerate changes in environmental conditions. Further more, Keller et al. , in order to preserve these species it is critical to understand their population dynamics and their habitat conservation status. Eucranium is characterized by its unusual morphology and unique biology and behavior, and it constitutes an old evolutionary lineage. Vane-Wright et al. (Eucranium of high conservation value.Species of lic loss . Accordir et al. , based ot et al. proposed"} +{"text": "Each of the two independent AuIII ions lies on an inversion center and has a distorted square-planar geometry. In the crystal, inter\u00admolecular C\u2014H\u22efCl hydrogen bonds, \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.5548\u2005(16) and 3.7507\u2005(16)\u2005\u00c5] and Au\u22ef\u03c0 inter\u00adactions [Au\u22efcentroid distance = 3.6424\u2005(10)\u2005\u00c5] are effective in the stabilization of the structure, resulting in the formation of a supra\u00admolecular structure. Intra\u00admolecular N\u2014H\u22efN hydrogen bonds are present in the cation.In the title compound, (C DOI: 10.1107/S1600536811036208/hy2466Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II ion in the title complex, [PtCl2(C14H10N4)], is four-coordinated in a distorted square-planar environment by two N atoms of a chelating 2,3-di-2-pyridyl\u00adpyrazine ligand and two chloride anions. The pyridyl ring coordinated to the PtII atom is inclined slightly to its carrier pyrazine ring [dihedral angle = 13.5\u2005(1)\u00b0], whereas the uncoordinated pyridyl ring is inclined considerably to the pyrazine ring [dihedral angle = 54.3\u2005(2)\u00b0]. The dihedral angle between the two pyridyl rings is 59.2\u2005(2)\u00b0. In the crystal, the complexes are assembled through inter\u00admolecular C\u2014H\u22efN and C\u2014H\u22efCl hydrogen bonds, forming a three-dimensional network. Intra\u00admolecular C\u2014H\u22efN and C\u2014H\u22efCl hydrogen bonds are also present.The Pt DOI: 10.1107/S1600536811038906/tk2794Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The NO5 donor set is based on a distorted octa\u00adhedron. Helical supra\u00admolecular chains along [010] are found in the crystal structure mediated by O\u2014H\u22efO hydrogen bonds formed between the coordinating methanol mol\u00adecule and the phenolate O atom of the chloro\u00adbenzene residue.The title Schiff base complex, [V(C E)-2-(2-hy\u00addroxy\u00adbenzyl\u00adidene\u00adamino)\u00adphenolates containing halide atoms on the aromatic ring(s), see: Yeni\u015fehirli et al. (CH3O)O(CH4O)] = 0.046wR(F2) = 0.131S = 1.043453 reflections221 parameters1 restraintH atoms treated by a mixture of independent and constrained refinementmax = 1.37 e \u00c5\u22123\u0394\u03c1min = \u22120.93 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 global, I. DOI: 10.1107/S160053681200311X/hg5167Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atoms in the title complex, [Ni2(CN)4(C16H36N4)]\u00b7C3H7NO\u00b70.5H2O, are bridged by a cyanide ion. The macrocycle folds around one NiII atom, which is five-coordinated in an NiN5 square-pyramidal geometry. The other NiII atom is surrounded by the cyanide ions in an NiN4 square-planar geometry. The dimethyl\u00adformamide solvent mol\u00adecule is disordered over two positions in a 0.62\u2005(1):0.38\u2005(1) ratio and the water mol\u00adecule is disordered about a center of inversion. The dinuclear mol\u00adecule and solvent mol\u00adecules are linked by N\u2014H\u22efO, N\u2013H\u22efN and O\u2014H\u22efO hydrogen bonds, forming a three-dimensional network.The two Ni DOI: 10.1107/S1600536810042625/bt5382Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Activation of the \u03b1-secretase processing pathway of amyloid precursor protein (APP) is recognized as an important mechanism which diverts APP processing from production of beta-amyloid (A\u03b2) to non toxic sAPP\u03b1, decreasing Alzheimer\u2019s disease (AD) plaque formation and AD-associated cognitive deficits. Two potent classes of PKC modulators can activate the \u03b1-secretase pathway, the benzo/indolactams and bryostatin/bryologues. While both modulate PKC-dependent APP processing, no direct comparisons of their relative pharmacological potencies have been accomplished which could assist in the development of AD therapies. In this study, we measured the activation of \u03b1-secretase APP processing and PKC-\u03b1, -\u03b4, and -\u03b5 induced by the benzolactam-APP modulator TPPB and bryostatin-1 in the neuroblastoma cell line SH-SY5Y which expresses APP and \u03b1- and \u03b2-secretase processing mechanisms. Bryostatin-1 produced a more rapid, potent, and sustained activation of \u03b1-secretase APP processing than TPPB and selectively activated PKC-\u03b4 and PKC-\u03b5. Although TPPB also activated \u03b1-secretase, its potency was approximately 10- to 100-fold lower, possibly reflecting lower PKC-\u03b4 and -\u03b5 activation. Because bryostatin-1 is a highly potent PKC-\u03b4 and -\u03b5 activator which activates \u03b1-secretase APP processing, further characterization of bryostatin-1/bryologues may help refine their use as important tools for the clinical management of AD. N-methyl-d-aspartate receptor antagonist, which reduces glutamate-dependent excitotoxicity, Ca2+ influx, and oxidant stress in neurons , few effective treatment options have been developed and \u201cstatins,\u201d inhibitors of HMG-coA reductase, may also reduce formation and persistence of A\u03b2 and have been suggested as potential adjuvant therapies for AD include diphenylurea, derivatives of hydroxyethylamine, and celastol, all of which at least partly block conversion of APP to A\u03b2. \u03a4hese drugs have been extensively studied in AD therapy , a disintegrin and metalloprotease (\u2018ADAM\u2019)-17 activation. Several drugs which modulate PKC isoenzyme activity, particularly the PKC-\u03b4 and PKC-\u03b5 isoforms, potently activate \u03b1-secretase processing of APP, generating the non toxic sAPP\u03b1 and preventing A\u03b2 formation. Currently, PKC activators may represent the most important, novel, and specific treatments for AD. At least two important classes of agents can activate PKC-dependent APP processing at relatively low (\u03bcM to nM) doses: (1) the benzolactam/indolactams and (2) bryostatin-1 and related bryologues.Ki\u2009=\u200911.9\u00a0nM for inhibition of phorbol 12,13 dibutryrate binding to PKC-\u03b1) which enhances sAPP\u03b1 secretion in fibroblasts and PC12 neuronal cells --8-(5-(4-(trifluoromethyl) phenyl)-2,4-pentadienoylamino) benzolactam] (CAS#497259-23-1) was purchased from Reagent 4 Research LLC and was diluted in ethanol. All ethanol concentrations in this study were below 0.1\u00a0% in cellular assays.Bryostatin-1 in ethanol stock solutions was provided by Aphios Corporation . The benzolactam APP modulator TPPB , -\u03b4 , and -\u03b5 antibodies . H-7 antibody is a mouse monoclonal antibody raised against amino acids 645\u2013672 on human PKC-\u03b1 which recognizes a single 80-kDA PKC-\u03b1 isoform. G-9 antibody is a mouse monoclonal antibody raised against amino acids 647\u2013673 of rat PKC-\u03b4 which recognizes a single 78-kDA band, and E-5 antibody is a mouse monoclonal antibody which was raised against amino acids 705\u2013737 of human PKC-\u03b5 and recognizes a single 90-kDA PKC-\u03b5 band. PKC isoform bands were visualized using HRP-conjugated 2\u00b0 antibody with ECL plus with SRX101 film. Band intensities were quantified by densitometric analysis using an HP Scanjet 3970 Densitometer and Image-J image analysis system software .Measurements of cytosolic and membrane-associated levels of PKC-\u03b1, -\u03b4, and -\u03b5 isoenzymes were used to assess PKC translocation in response to bryostatin-1 or TPPB as described by Racchi et al. with modp\u2009<\u20090.05 was considered as statistically significant.All data are presented as mean \u00b1 standard deviation and were analyzed by one-way ANOVA with Dunnett\u2019s post-testing . A value of \u22127\u00a0M (**p\u2009<\u20090.01), 10\u22128\u00a0M (**p\u2009<\u20090.01), 10\u22129\u00a0M (*p\u2009<\u20090.05), and 10\u221210\u00a0M (*p\u2009<\u20090.05). TPPB also induced sAPP\u03b1 release, but to a lesser extent and only at 10\u22127\u00a0M (p\u2009<\u20090.05) and 10\u22128\u00a0M (p\u2009<\u20090.05) ; TPPB was able to significantly increase sAPP\u03b1 at 6\u00a0h (*p\u2009<\u20090.05) , 6\u00a0h (**p\u2009<\u20090.01), and 12\u00a0h (*p\u2009<\u20090.05). At this dose, TPPB only significantly increased the release of sAPP\u03b1 at 6\u00a0h (*p\u2009<\u20090.05) , which persisted at 1\u00a0h (**p\u2009<\u20090.01) and 3\u00a0h time points (**p\u2009<\u20090.01). By comparison, TPPB only induced the significant release of sAPP\u03b1 at 3\u00a0h (*p\u2009<\u20090.05) ; however, TPPB at 10\u221210\u00a0M did not significantly increase the release of sAPP\u03b1 at any of the time points tested Fig.\u00a0. At 10\u2212805) Fig.\u00a0. Bryosta05) Fig.\u00a0. Bryostated Fig.\u00a0.Fig. 2a\u22128, 10\u22129, and 10\u221210\u00a0M bryostatin-1 significantly increased the activation of PKC-\u03b1 in SH-SY5Y cells (**p\u2009<\u20090.01) ; 10\u221210\u00a0M TPPB did not activate PKC-\u03b1 at any time point tested. A dose-dependent comparison of bryostatin-1 and TPPB showed that, at 3\u00a0h, PKC-\u03b4 was activated by 10\u22128\u00a0M bryostatin-1 (**p\u2009<\u20090.01), 10\u22129\u00a0M bryostatin-1 (*p\u2009<\u20090.05), and 10\u221210\u00a0M bryostatin-1 (*p\u2009<\u20090.05) , 1\u00a0h (**p\u2009<\u20090.01), and 3\u00a0h (**p\u2009<\u20090.01); by comparison, 10\u22129\u00a0M TPPB did not activate PKC-\u03b4 at any time point tested and 3\u00a0h (**p\u2009<\u20090.01); by comparison, 10\u221210\u00a0M TPPB did not activate PKC-\u03b4 at any time point , 10\u22129\u00a0M bryostatin-1 (**p\u2009<\u20090.01), and 10\u221210\u00a0M bryostatin-1 (*p\u2009<\u20090.05) , 1\u00a0h (**p\u2009<\u20090.01), and 3\u00a0h (**p\u2009<\u20090.01) , 1\u00a0h (*p\u2009<\u20090.01), and 3\u00a0h (**p\u2009<\u20090.01) following treatment with bryostatin-1 or TPPB. Confluent cultures of SH-SY5Y neuroblastoma cells were treated with different concentrations of either bryostatin-1 or TPPB for time points up to 3\u00a0h, when membrane and cytosolic pools of PKCs were isolated by differential detergent extraction, and Western-blotted for each individual isoform. Actin was also Western-blotted in each fraction to ensure equal loading. At 3\u00a0h, 1001) Fig.\u00a0; 10\u22128, 1C-\u03b1 Fig.\u00a0. A time 01) Fig.\u00a0; 10\u22129\u00a0M 01) Fig.\u00a0. Bryosta05) Fig.\u00a0; TPPB di05) Fig.\u00a0. A time ted Fig.\u00a0. A time int Fig.\u00a0. A dose-05) Fig.\u00a0; TPPB di05) Fig.\u00a0. A time 01) Fig.\u00a0; by compted Fig.\u00a0. A time 01) Fig.\u00a0; by compted Fig.\u00a0.Fig. 3aThe presence of A\u03b2-laden cerebral plaques in AD, a central feature of Alzheimer\u2019s disease (AD), is thought to contribute to the disturbances in nerve structure, function, and memory which characterize AD dementia GABA(A)R-regulated post-synaptic currents, and (3) cholinergic signaling activates \u03b1-secretase via PKC-\u03b1 and \u201cnovel\u201d PKC isoforms to stimulate sAPP\u03b1 processing to enhance non-amyloidogenic processing of APP to sAPP\u03b1. Currently, the toxicological properties of TPPB are not fully known. An important goal in PKC\u2013APP modulator drug design must be to overcome the potential carcinogenic properties of benzolactams/indolactams because TPPB, like TPA and other phorbol esters, may still retain tumor-promoting activity of TPPB needed to reach equivalent \u03b1-secretase activation as bryostatin-1 which may hamper its use in human AD therapy. There are, of course, relative factors of availability and cost of bryostatin-1 versus reduced potency and higher toxicity of TPPB which can ultimately determine clinical use.By comparison, TPPB is a novel, synthetic cell-permeable benzolactam PKC activator which has been used as a standard APP modulator . Bryostatin-1 also appears to be a more active, rapid, and persistent APP modulator which reflects its greater specificity toward PKC-\u03b4 or PKC-\u03b5 isoform activation at low doses.Novel PKC activators are important candidate treatments for AD because they activate beneficial APP processing through \u03b1-secretase. We compared how PKC-\u03b1 , -\u03b4, and -\u03b5 (novel PKC isoforms) were activated in neuroblastoma cells by bryostatin-1 and TPPB as a function of time and concentration. Bryostatin-1 significantly induced PKC-\u03b1, -\u03b4, and -\u03b5 activation at 10Here, we found that bryostatin-1 exerted a more rapid, potent, and sustained activation of APP processing , which was associated with its more potent and specific activation of PKC-\u03b4 and PKC-\u03b5 at low doses. The further refinement of bryostatin-1 pharmacology in PKC mobilization and \u03b1-secretase activation may help move this drug class more rapidly into clinical treatment for AD."} +{"text": "The pyridyl N atom of the Schiff base and the imino N atom of the 4-methyl-pyridine-2-yl\u00adimino ligand are not involved in the coordination. There is an intra\u00admolecular N\u2014H\u22efN hydrogen bond involving the pyridine N atom and the amino group of the 2-amino\u00adpyridine ligand. In the crystal, mol\u00adecules are linked via N\u2014H\u22efCl hydrogen bonds, forming chains propagating along [001].In the title complex, [Cu(C Cl(C6H8N2) = 0.055wR(F2) = 0.114S = 0.943339 reflections243 parameters2 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.40 e \u00c5\u22123\u0394\u03c1min = \u22120.45 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536812047198/su2518Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The carboxyl\u00adate groups of the HB ions are slightly twisted away from the attached benzene rings by 2.84\u2005(15) and 4.8\u2005(2)\u00b0. The planes of the two benzene rings of the HB ions are oriented with respect to each other at a dihedral angle of 84.41\u2005(8)\u00b0. In the crystal, adjacent polymeric chains inter\u00adact via O\u2014H\u22efO, N\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds. The solvent water mol\u00adecule links with the polymeric chains via O\u2014H\u22efO hydrogen bonding. \u03c0\u2013\u03c0 stacking between the benzene and pyridine rings and between the benzene rings [centroid\u2013centroid distances = 3.731\u2005(2) and 3.353\u2005(2)\u2005\u00c5] are present in the crystal.In the crystal of the title polymeric compound, {[Pb(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)]\u00b7H2O = 0.017wR(F2) = 0.050S = 1.194783 reflections304 parameters4 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.94 e \u00c5\u22123\u0394\u03c1min = \u22121.27 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812004357/xu5463Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atom in the polymeric title compound, {[Pb(C13H11N4O)]ClO4}n, is coordinated by the N\u2032-[1-(pyridin-2-yl-\u03baN)ethyl\u00adidene]isonicotinohydrazidate ligand via its O,N,N\u2032-donors and simultaneously bridged by a neighbouring ligand via the pyridin-2-yl N atom. The resultant supra\u00admolecular chain is a zigzag along the a axis. The stereochemistry of the PbII atom is defined by an N3OE donor set (E = lone pair of electrons), which results in a \u03a8-trigonal\u2013bipyramidal coordination with the O and pyridin-2-yl N atoms in axial positions. The dihedral angle between the pyridine rings of the ligand is 6.3\u2005(3)\u00b0. The supra\u00admolecular cationic chains are linked into a three-dimensional array via secondary Pb\u22efO [3.133\u2005(6) and 3.28\u2005(7)\u2005\u00c5] and Pb\u22efN [3.028\u2005(4)\u2005\u00c5] inter\u00adactions. Weak C\u2014H\u22efO inter\u00adactions and aromatic \u03c0\u2013\u03c0 stacking [centroid\u2013centroid separation = 3.693\u2005(2)\u2005\u00c5] also occur in the crystal.The Pb N\u2032-[1-(2-pyrid\u00adyl)ethyl\u00adidene]isonicotinohydrazide ligand, see: Maurya et al. ]ClO4 = 0.022 wR(F 2) = 0.056 S = 1.07 2811 reflections218 parametersH-atom parameters constrainedmax = 0.55 e \u00c5\u22123 \u0394\u03c1min = \u22120.45 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811046691/hb6465Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecules are linked by O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds. Two types of \u03c0\u2013\u03c0 stacking inter\u00adactions occur between the TPTZ ligands of adjacent complexes [centroid-to-centroid distances = 3.760\u2005(4) and 3.870\u2005(3)\u2005\u00c5].In the title compound, [Nd(NO DOI: 10.1107/S1600536811014589/ff2007Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One of the CuII ions is coordinated in a distorted square-planar geometry, whereas the other is coordinated in a distorted square-pyramidal geometry, the long apical Cu\u2014O bond [2.885\u2005(4)\u2005\u00c5] of the square-pyramidal coordination being provided by a symmetry-related O atom creating a one-dimensional polymer along [010]. \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.783\u2005(4)\u2005\u00c5] and short inter\u00adchain Br\u22efBr inter\u00adactions [3.6142\u2005(12)\u20133.6797\u2005(12)\u2005\u00c5] are observed.The asymmetric unit of the title coordination polymer consists of a dinuclear neutral complex mol\u00adecule of formula [Cu DOI: 10.1107/S1600536812029285/rz2776Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Adjacent complex mol\u00adecules are stacked through aromatic \u03c0\u2013\u03c0 inter\u00adactions; the closest distance between adjacent aromatic rings is 3.598\u2005(2)\u2005\u00c5.In the mononuclear title compound, [ZnCl DOI: 10.1107/S1600536812022313/rk2337Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The four O atoms in the equatorial plane around the CoII cation form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 23.91\u2005(9)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 88.84\u2005(4)\u00b0. The coordinating water mol\u00adecule links with the carboxyl\u00adate group via an intra\u00admolecular O\u2014H\u22efO hydrogen bond. In the crystal, N\u2014H\u22efO, O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular network. \u03c0\u2013\u03c0 stacking between the parallel benzene rings of adjacent mol\u00adecules [centroid\u2013centroid distance = 3.8505\u2005(8)\u2005\u00c5] may further stabilize the structure. A weak C\u2014H\u22ef\u03c0 inter\u00adaction also occurs in the crystal.In the title complex, [Co(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.026wR(F2) = 0.069S = 1.053527 reflections216 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.42 e \u00c5\u22123\u0394\u03c1min = \u22120.36 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812032205/xu5597Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "This is only possible as the mol\u00adecule has a hinged cis arrangement about the Pd2+ coordination spheres. For this hinged dimeric structure, the angles between the two coordination planes in each mol\u00adecule are 21.59\u2005(4) and 22.10\u2005(4)\u00b0. This hinged cis arrangement also allows the two mol\u00adecules to form pairs linked by secondary inter\u00adactions between the Pd and Te atoms of an adjoining mol\u00adecule, leading to a tetra\u00admeric overall structure. C\u2014H\u22efCl inter\u00adactions consolidate the crystal packing.The asymmetric unit of the title compound, [Pd For related structures of bridged dimers of palladium mediated by Se, see: Brown & Corrigan 2004; Chakrab al. 2011; Dey et al. 2006; Ford et al. 2004; Kaur et al. 2009; Morley al. 2006; Nakata al. 2009; Oilunka al. 1999, 2001 \u25b6.2(C9H12NTe)2Cl2]\u00b70.5CH2Cl2 = 0.039wR(F2) = 0.102S = 1.065506 reflections254 parametersH-atom parameters constrainedmax = 2.12 e \u00c5\u22123\u0394\u03c1min = \u22120.90 e \u00c5\u22123\u0394\u03c1Absolute structure: Flack 1983, 2355 FrFlack parameter: 0.06 (4)APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812000104/jj2116Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CuII atom lies 0.065\u2005(1)\u2005\u00c5 above the N2O2 plane and the Cu\u2014O [2 \u00d7 1.945\u2005(2)\u2005\u00c5] and Cu\u2014N bond lengths [1.968\u2005(3) and 1.962\u2005(3)\u2005\u00c5] lie in expected ranges. The two aromatic ring planes make a dihedral angle of 85.48\u2005(1)\u00b0.The mol\u00adecular structure of the title compound, [Cu(C DOI: Click here for additional data file.10.1107/S1600536812046387/aa2076Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The metal-coordinated pyridine rings are almost coplanar, making a dihedral angle of 1.5\u2005(2)\u00b0, while the two pendent pyrazine rings are arranged on the same side of the N\u2014Ag\u2014N line. Along the a axis, the mononuclear coordination units are stacked with \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings [centroid\u2013centroid distance = 3.569\u2005(4)\u2005\u00c5], leading to infinite chains. The chains are inter\u00adconnected through inter\u00admolecular N(pyrazine)\u22ef\u03c0(pyrazine) inter\u00adactions forming layers parallel to the ab plane [N\u22efcentroid = 3.268\u2005(5)\u2005\u00c5]. These layers are further stacked along the c-axis direction, furnishing a three-dimensional supra\u00admolecular framework with the tetra\u00adfluoridoborate anions embedded within the inter\u00adstices.In the title mononuclear complex, [Ag(C DOI: 10.1107/S1600536811051270/zq2141Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the complex cations and the monodeprotonated 2-carb\u00adoxy\u00adbenzoate anions are connected by O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming a tape along [100]. Adjacent tapes are further linked into a three-dimensional arrangement via \u03c0\u2013\u03c0 stacking inter\u00adactions between the triazole and benzene rings and between the pyridine and benzene rings [centroid\u2013centroid distances = 3.6734\u2005(14)/3.9430\u2005(16) and 3.8221\u2005(14)\u2005\u00c5]. Intra\u00admolecular N\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds are also observed.In the complex cation of the title salt, [Cu(C DOI: 10.1107/S1600536812000128/hy2503Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The coordination geometry around the CuII atom is square-pyramidal, comprising two N atoms from a symmetrically chelating 1,10-phenanthroline ligand, one O atom from a trichloro\u00adacetate ligand and two Cl\u2212 anions. In addition, there is a weak intra\u00admolecular Cu\u22efO inter\u00adaction of 2.9403\u2005(14)\u2005\u00c5 involving the carbonyl O atom of the trichloro\u00adacetate ligand. The central Cu2Cl2 core takes the form of a rhombus, owing to the disparate Cu\u2014Cl bond lengths. Mol\u00adecules are connected in the crystal structure by C\u2014H\u22efCl and C\u2014H\u22efO inter\u00adactions.The title compound, [Cu II 1,10-phenanthroline complexes, see: De Burgomaster et al. 2Cl2(C12H8N2)2] = 0.026wR(F2) = 0.070S = 1.053233 reflections208 parametersH-atom parameters constrainedmax = 0.52 e \u00c5\u22123\u0394\u03c1min = \u22120.46 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812003947/bt5807Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, each palladium dimer is accompanied by a dichloro\u00admethane solvent mol\u00adecule. Coordination of the carbonate and chelated phosphane ligands gives distorted square-planar environments at the Pd atoms. Important geometrical parameters include Pd\u2014P(av.) = 2.2135\u2005(4)\u2005\u00c5, Pd\u2014C(av.) = 1.9648\u2005(16)\u2005\u00c5 and P\u2014Pd\u2014C = 84.05\u2005(5) and 87.98\u2005(5)\u00b0, and O\u2014Pd\u2014O\u2032 = 60.56\u2005(4) and 61.13\u2005(4)\u00b0. Bonding with the carbonate O atoms shows values of 2.1616\u2005(11) and 2.1452\u2005(11)\u2005\u00c5 for the Pd\u2014O\u2014Pd bridge, whereas other Pd\u2014O distances are slightly longer at 2.2136\u2005(11) and 2.1946\u2005(11)\u2005\u00c5. One of the tert-butyl groups is disordered over two set of sites with an occupancy ratio of 0.723\u2005(6):0.277\u2005(6). Weak C\u2014H\u22efO interactions are observed propagating the molecules along the [100] direction. The title compound, [(\u03bc DOI: Click here for additional data file.10.1107/S1600536812048702/yk2080Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "A chain along the c axis is formed by C\u2014H\u22efN hydrogen-bonding inter\u00adactions and a weak \u03c0\u2013\u03c0 inter\u00adaction is observed between the pyrimidine rings of two adjacent parallel chains [centroid\u2013centroid distance = 3.722\u2005(2)\u2005\u00c5]. N\u2014H\u22efCl, CN\u2014H\u22efCl and N\u2014H\u22efO interactions also occur. In the title compound, (C DOI: 10.1107/S1600536811045995/vn2019Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The water mol\u00adecules are involved in intra- and inter\u00admolecular O\u2014H\u22efO and O\u2014H\u22efCl hydrogen bonding. Inter\u00admolecular N\u2014H\u22efO and N\u2014H\u22efCl hydrogen bonds are formed between ligands. In addition, weak \u03c0\u2013\u03c0 inter\u00adactions are observed between the benzene rings of the ligands [centroid\u2013centroid distance = 3.580\u2005(3)\u2005\u00c5]. The inter\u00admolecular hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions lead to the formation of a three-dimensional supra\u00admolecular network.In the title complex, [NiCl H)-one, see: Turgunov & Englert (2010H)-one, see: Turgunov et al. (2010For a Cd(II) coordination polymer with quinazolin-42(H2O)2] = 0.052wR(F2) = 0.139S = 0.941686 reflections132 parameters2 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.75 e \u00c5\u22123\u0394\u03c1min = \u22120.51 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812019381/hg5221Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Extensive inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds lead to the stability of the crystal structure. Inter\u00adactions between one C\u2014H group of the 2,3-diamino\u00adpyridinium [+] cation and the aromatic ring of the pyridine-2,6-dicarboxyl\u00adate (pydc) ligand (C\u2014H\u22efcentroid distance = 2.78\u2005\u00c5) and \u03c0\u2013\u03c0 inter\u00adactions between the + cations and between the + cation and the pydc ligand [centroid\u2013centroid distances = 3.489\u2005(5) and 3.694\u2005(5)\u2005\u00c5] are observed.In the centrosymmetric dinuclear complex anion of the title compound, (C DOI: 10.1107/S1600536811005629/hy2406Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine and benzene rings of adjacent mol\u00adecules, with a centroid\u2013centroid distance of 3.829\u2005(2)\u2005\u00c5.In the title centrosymmetric dinuclear complex, [Eu DOI: 10.1107/S1600536810046131/hy2358Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "III cations in the polymeric title compound, {[La2(C7H3NO4)3(H2O)4]\u00b72H2O}n. One is nine-coordinated in an LaN2O7 tricapped trigonal\u2013prismatic geometry formed by three pyridine-2,6-dicarboxyl\u00adate anions and two water mol\u00adecules, while the other is ten-coordinated in an LaNO9 bicapped square-anti\u00adprismatic geometry formed by four pyridine-2,6-dicarboxyl\u00adate anions and two water mol\u00adecules. The two LaIII cations are separated by a non-bonding distance of 5.026\u2005(3)\u2005\u00c5. The pyridine-2,6-dicarboxyl\u00adate anions bridge the LaIII cations, forming a three-dimensional polymeric complex. The crystal structure contains extensive classical O\u2014H\u22efO hydrogen bonds and weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds. The crystal structure is further consolidated by \u03c0\u2013\u03c0 stacking between pyridine rings, the shortest centroid\u2013centroid distance between parallel pyridine rings being 3.700\u2005(5)\u2005\u00c5.There are two independent La DOI: 10.1107/S1600536811030807/xu5269Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "It is coordinated by two water mol\u00adecules in the apical directions and four 4-fluoro\u00adbenzoate (PFB) anions, bridging the symmetry related Mn atoms in the basal plane to form an infinite two-dimensional polymeric structure parallel to (100). The four O atoms of the PFB anions around the MnII atom form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two O atoms of the water mol\u00adecules. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 27.29\u2005(16)\u00b0. The O\u2014H\u22efO hydrogen bonds further connect the manganese-carboxyl\u00adate units. \u03c0\u2013\u03c0 contacts between the benzene rings [centroid-centroid distance = 3.6894\u2005(15)\u2005\u00c5] further stabilize the crystal structure.In the crystal structure of the title complex, [Mn(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(H2O)2] = 0.039 wR(F 2) = 0.111 S = 1.28 1758 reflections114 parametersH atoms treated by a mixture of independent and constrained refinementmax = 1.24 e \u00c5\u22123 \u0394\u03c1min = \u22120.45 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811021921/su2282Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordination network can be described as an uninodal 4-connected net with the sql topology. The HgII ion lies on a site of -1 symmetry and the apym ligand lies on sites of m symmetry with the mirror plane perpendicular to the pyrimidine plane and passing through the NH2 group N atom. This polymeric structure is stabilized by N\u2014H\u22efCl hydrogen bonds and columnar \u03c0\u2013\u03c0 stacking of pyrimidine rings, with a centroid\u2013centroid distance of 3.832\u2005(2)\u2005\u00c5.The title compound, [HgCl For mer al. 2007. For the al. 2007; Xie & W al. 2007. For top al. 2007. For an al. 2011. For our al. 2009, 2011 \u25b6.2(C4H5N3)] = 0.020wR(F2) = 0.049S = 1.12992 reflections52 parametersH-atom parameters constrainedmax = 0.84 e \u00c5\u22123\u0394\u03c1min = \u22121.01 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812008793/gk2458Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II ion in the title compound, [Mn(CH3CO2)2(C15H11N3)]\u00b72H2O, is seven-coordinated in a considerably distorted penta\u00adgonal\u2013bipyramidal geometry by three N atoms of the tridentate 2,2\u2032:6\u2032,2\u2032\u2032-terpyridine ligand and four O atoms from two acetate anions which chelate the Mn atom via two O atoms. The lateral pyridine rings are slightly inclined to the central pyridine ring, making dihedral angles of 13.6\u2005(2) and 5.7\u2005(2)\u00b0. The complex and solvent water mol\u00adecules are linked by inter\u00admolecular O\u2014H\u22efO hydrogen bonds into a three-dimensional network.The Mn For rel al. 2004; Rich et al. 2010.2H3O2)2(C15H11N3)]\u00b72H2O = 0.055wR(F2) = 0.146S = 0.934936 reflections276 parameters4 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.69 e \u00c5\u22123\u0394\u03c1min = \u22120.52 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The IrIII atom is coordinated by an \u03b75-C5Me5 ligand, a chloride and a Ph2P(CH2)3SPh-\u03baP,\u03baS ligand, leading to a three-legged piano-stool geometry. In the crystal, two water molecules and two chloride anions are linked by weak O\u2014H\u22efCl hydrogen bonding into tetra\u00admers that are located on centers of inversion. The H atoms of one of the methyl groups are disordered and were refined using a split model.The crystal structure of the title compound, [Ir(C For an al. 2012.10H15)Cl(C21H21PS)]Cl\u00b7H2O = 0.041wR(F2) = 0.088S = 1.255513 reflections337 parameters147 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 1.68 e \u00c5\u22123\u0394\u03c1min = \u22121.50 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812021964/nc2278Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "An intra\u00admolecular N\u2014H\u22efO(carbon\u00adyl) hydrogen bond closes an S(6) ring. Supra\u00admolecular chains along [01-1] mediated by O\u2014H\u22efN(pyridine) hydrogen bonds feature in the crystal. A three-dimensional architecture is completed by \u03c0\u2013\u03c0 inter\u00adactions occurring between the benzene ring and the two rings of the thienopyridine residue [centroid\u2013centroid distances = 3.6963\u2005(13) and 3.3812\u2005(13)\u2005\u00c5]. The F atom is disordered over the two meta sites in a near statistical ratio [0.545\u2005(5):0.455\u2005(5)].In the title compound, C DOI: 10.1107/S1600536812027195/hb6844Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S1600536812027195/hb6844Isup3.cmlSupplementary material file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the Me\u2013salen ligand, the benzene rings are almost parallel, making a dihedral angle of 0.48\u2005(13)\u00b0, but the torsion angle along the central C\u2014C bond is 41.1\u2005(2)\u00b0\u00b7The pyrrolidine rings are in slightly distorted chair conformations. The N atoms of the pyrrolidine axial ligands are involved in N\u2014H\u22efO hydrogen bonds with the perchlorate anions, and these hydrogen bonds connect the ionic species into infinite chains along the b axis. Some relatively short C\u2014H\u22ef\u03c0 inter\u00adactions are also present in the crystal structure and C\u2014H\u22efO inter\u00adactions occur. The guest solvent p-xylene mol\u00adecule lies on a special position at the inversion centre.In the mononuclear title complex, [Co(C For rel al. 2003. For thebach 1969. 18H18N2O2)(C4H9N)2]ClO4\u00b70.5C8H10 = 0.040 wR(F 2) = 0.084 S = 0.99 7081 reflections502 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.67 e \u00c5\u22123 \u0394\u03c1min = \u22120.51 e \u00c5\u22123 \u0394\u03c1 CrysAlis PRO used to solve structure: SIR92 (Altomare et al., 1993SHELXL97 (Sheldrick, 2008Stereochemical Workstation Operation Manual (Siemens, 1989SHELXL97.Data collection: 10.1107/S160053681004660X/jh2227sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S160053681004660X/jh2227Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CeIII atom is coordinated by eight O atoms from six INAIP ligands and a water mol\u00adecule in a distorted tricapped trigonal\u2013prismatic geometry, while the AgI atom has a distorted trigonal\u2013planar AgN2O geometry. O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the pyridine and benzene rings [centroid\u2013centroid distances = 3.642\u2005(4) and 3.624\u2005(3)\u2005\u00c5] stabilize the crystal structure.The 4 DOI: 10.1107/S1600536811027668/hy2446Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The equatorial Cu\u2014N and Cu\u2014O bond lengths are in the range 1.979\u2005(2)-1.998\u2005(3)\u2005\u00c5. The axial Cu\u2014O bond distances are 2.365\u2005(2) and 2.394\u2005(2)\u2005\u00c5. In the crystal, the complex cations and perchlorate anions are connected by numerous C\u2014H\u22efO hydrogen bonds, which leads to additional stabilization of the structure. The perchlorate anion is disordered over two sets of sites with a 0.716\u2005(3):0.284\u2005(3) occupancy ratio.The title compound, [Cu(C DOI: 10.1107/S1600536813018485/nr2046Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Ag\u2014C and Ag\u2014Cl single-bond lengths are 2.087\u2005(3) and 2.3267\u2005(9)\u2005\u00c5. The C\u2014Ag\u2014Cl bond angle is 172.84\u2005(7)\u00b0. C\u2014H\u22ef\u03c0 inter\u00adactions contribute to the stabilization of the crystal structure. A very weak \u03c0\u2013\u03c0 stacking inter\u00adaction between adjacent tetra\u00admethyl\u00adbenzene rings [centroid\u2013centroid distance = 4.0610\u2005(18)\u2005\u00c5] is also observed.In the title compound, [AgCl(C DOI: 10.1107/S1600536812012998/sj5223Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Each AgI ion is bicoordinated in a slightly distorted linear coordination geometry by the N atoms of two ligands, resulting in the formation of a 22-membered metallamacrocycle. In the dication, \u03c0\u2013\u03c0 inter\u00adactions are observed between the imidazole rings, with centroid\u2013centroid distances of 3.528\u2005(3)\u2005\u00c5 and dihedral angles of 9.92\u2005(9)\u00b0. The crystal structure is stabilized by inter\u00admolecular O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions involving the benzene rings of adjacent dications, with centroid\u2013centroid distances of 3.651\u2005(2)\u2005\u00c5.In the title compound, [Ag DOI: 10.1107/S1600536811018691/rz2589Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CoII atom is situated on a crystallographic center of inversion and is octa\u00adhedrally coordinated by two O atoms from two anions, two N atoms of two 1,2-bis\u00ad(pyridin-4-yl)ethane mol\u00adecules and two O atoms from two water mol\u00adecules. A three-dimensional network is established by inter\u00admolecular O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds.The asymmetric unit of the title compound, [Co(C For general background to the design of metal-organic supra\u00admolecular solids with potential functionality, see: Moulton & Zaworotko 2001; Janiak 9H7O4)2(C12H10N2)2(H2O)2] = 0.055 wR(F 2) = 0.105 S = 1.06 3635 reflections268 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.71 e \u00c5\u22123 \u0394\u03c1min = \u22120.36 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811024755/im2300Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII atom lies on a crystallographic twofold rotation axis and shows a significantly distorted tetra\u00adhedral coordination geometry. The dihedral angle between the phenyl rings is 74.3\u2005(2)\u00b0. The crystal structure is stabilized by inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.635\u2005(2)\u20133.803\u2005(3)\u2005\u00c5].The asymmetric unit of the title compound, [CuCl DOI: 10.1107/S1600536811050690/rz2665Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 contacts between the pyridine rings [centroid\u2013centroid distance = 3.510\u2005(3)\u2005\u00c5] are present in the crystal.In the title compound, [InCl DOI: 10.1107/S1600536812038147/hy2584Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "One AgI ion is coordinated by one O atom from a 3-carboxyl\u00adato-4-hy\u00addroxy\u00adbenzene\u00adsulfonate (L) ligand, two N atoms from two pyrazine ligands and a water mol\u00adecule. The other AgI ion is coordinated by two O atoms from two L ligands and one N atom from a pyrazine ligand. One of the pyrazine ligands lies on an inversion center. The L and pyrazine ligands link the AgI ions into polymeric layers parallel to the ac plane. The layers are connected by inter\u00admolecular O\u2014H\u22efO hydrogen bonds. An intra\u00admolecular O\u2014H\u22efO hydrogen bond is also present in the L ligand.The title coordination polymer, {[Ag For a related structure, see: Nie & Qu 2011. 4(C7H4O6S)2(C4H4N2)3(H2O)2]\u00b72H2O = 0.033 wR(F 2) = 0.073 S = 0.88 3387 reflections256 parameters5 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.70 e \u00c5\u22123 \u0394\u03c1min = \u22121.09 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811041626/hy2474Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordination environment of each ReI atom is distorted octahedral. Four intra\u00admolecular O\u2014H\u22efN and four inter\u00admolecular C\u2014H\u22efO hydrogen-bond inter\u00adactions are observed. Relatively strong hydrogen bonds are found between the hydrogen-bond donor (\u03bc3-OH) and acceptor (basic N atom of pyridine), with N\u22efO distances between 2.586\u2005(10) and 2.628\u2005(10)\u2005\u00c5. Inter\u00adcube distances of 9.873\u2005(2) and 12.376\u2005(3)\u2005\u00c5 are observed.The title compound, [Re DOI: 10.1107/S1600536812036033/hg5243Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Each RhI atom is coordinated by one O atom [Rh\u2014O = 2.044\u2005(2) and 2.026\u2005(2)\u2005\u00c5], one N atom [Rh\u2014N = 2.083\u2005(2) and 2.090\u2005(2)\u2005\u00c5], and one COD ligand via two \u03b72-bonds, each directed toward the mid-point of a C=C bond (Cg): Rh\u2014Cg = 2.007\u2005(2), 2.013\u2005(2), 2.000\u2005(2) and 2.021\u2005(2)\u2005\u00c5. Each RhI atom has a quasi-square-planar coordination geometry, with average r.m.s. deviations of 0.159\u2005(1) and 0.204\u2005(1)\u2005\u00c5 from the mean planes defined by Rh and the termini of its four coordinating bonds. The two COD ligands have quasi-C2 symmetry, twisted from ideal Cv2 symmetry by 30.0\u2005(3) and \u221233.1\u2005(3)\u00b0, and are quasi-enanti\u00adomers of one another. The intra\u00admolecular Rh\u22efRh distance of 5.9432\u2005(3)\u2005\u00c5 suggests that there is no direct metal\u2013metal inter\u00adaction.In the title solvate, [Rh DOI: 10.1107/S1600536812040603/cv5334Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The water ligands of the core are involved in hydrogen bonds with the triazole N atoms of the organic mol\u00adecules, which generates a layer motif in the ab plane. There are \u03c0\u2013\u03c0 stacking inter\u00adactions between benzene rings of 3.490\u2005(6)\u2005\u00c5, and between triazole rings of 3.543\u2005(8) and 3.734\u2005(7)\u2005\u00c5 in neighboring layers, forming a three-dimensional network.The title complex, [Fe DOI: 10.1107/S160053681202613X/pk2413Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atom in the title salt, [Cu(C10H24N4)(H2O)2](C8H7O2)2\u00b7H2O, is chelated by the four N atoms of the 1,4,8,11-tetra\u00adaza\u00adcyclo\u00adtetra\u00addecane (cyclam) ligand and is coordinated by two water mol\u00adecules in a Jahn\u2013Teller-type of tetra\u00adgonally distorted octa\u00adhedral geometry. The cations, anions and lattice water mol\u00adecules are linked by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds to form a layer structure parallel to (001).The Cu DOI: 10.1107/S1600536810026012/bt5288Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In each mol\u00adecule, the fused six-membered rings have chair\u2013chair\u2013chair\u2013chair conformations and the cyclo\u00adpentane ring adopts an envelope conformation with the C atom bearing the hy\u00addroxy\u00admethyl group as the flap. All ring junctions are trans-fused. With the exception of one of the methyl groups adjacent to the C=O group, all the methyl groups are in axial positions. The isopropenyl group is equatorial and the hy\u00addroxy\u00admethyl group is in an axial orientation. In the crystal, weak C\u2014H\u22efO inter\u00adactions link the mol\u00adecules into chains along [010]. Weak intra\u00admolecular C\u2014H\u22efO hydrogen bonds are also observed but the hy\u00addroxy groups are not involved in hydrogen bonds.The asymmetric unit of the title compound, C \u00c5b = 19.1818 (6) \u00c5c = 28.1141 (7) \u00c5V = 5093.3 (3) \u00c53Z = 8K\u03b1 radiationMo \u22121\u03bc = 0.07 mmT = 100 K0.60 \u00d7 0.56 \u00d7 0.20 mmOxford Diffraction diffractometer with a Sapphire3 detectorCrysAlis RED; Oxford Diffraction, 2008Tmin = 0.960, Tmax = 0.986Absorption correction: multi-scan (61473 measured reflections5036 independent reflectionsI > 2\u03c3(I)4401 reflections with Rint = 0.055R[F2 > 2\u03c3(F2)] = 0.051wR(F2) = 0.141S = 1.035036 reflections609 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.55 e \u00c5\u22123\u0394\u03c1min = \u22120.50 e \u00c5\u22123\u0394\u03c1CrysAlis CCD used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536813011008/lh5603Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The pyridinium rings are almost planar [maximum deviations = 0.004\u2005(4) and 0.003\u2005(4)\u2005\u00c5]. The ethyl groups are approximately perpendicular to the corresponding pyridinium ring planes [N\u2014C\u2014C\u2014C = 88.8\u2005(4)\u00b0 in each ligand]. The packing of the mol\u00adecules is controlled by \u03c0\u2013\u03c0 inter\u00adactions, with centroid\u2013centroid distances of 3.625\u2005(3) and 3.711\u2005(2)\u2005\u00c5, forming chains approximately parallel to (102). The crystal studied was non-merohedrally twinned .In the bioactive title compound, [ZnBr DOI: 10.1107/S1600536810052190/sj5050Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Two O,O\u2032-bis\u00ad(4-methyl\u00adphen\u00adyl)dithio\u00adphosphate anions occupy the axial positions with long Cu\u22efS distances of 3.0090\u2005(8)\u2005\u00c5. Inter\u00admolecular N\u2014H\u22efS and C\u2014H\u22efS hydrogen bonding is present between the anions and the cation.In the title compound, [Cu(C DOI: 10.1107/S1600536810046672/xu5079Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Four symmetry-equivalent mesitylacetamidate ligands bridge the Rh\u2014Rh unit. Thus, each RhII atom has an approximately octa\u00adhedral coordination by one Rh [Rh\u2014Rh = 2.4290\u2005(6)\u2005\u00c5], two acetamidate O atoms trans to each other [Rh\u2014O = 2.044\u2005(3)\u2005\u00c5], two acetamidate N atoms trans to each other [Rh\u2014N = 2.091\u2005(4)\u2005\u00c5], and a benzonitrile N atom trans to Rh [Rh\u2014N = 2.222\u2005(3)\u2005\u00c5]. The structure is held together by weak van der Waals forces.The title structure, [Rh DOI: 10.1107/S1600536812024518/qk2033Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds, generating [001] C 2 2(4) chains such that mol\u00adecules A and B alternate. There is no aromatic \u03c0\u2013\u03c0 stacking in the crystal as the shortest centroid\u2013centroid distance is greater than 4.74\u2005\u00c5.The previous structure determination [Gillier-Pandraud al. 1972. C. R. A DOI: 10.1107/S1600536811013547/hb5838Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II(TpivPP) {TpivPP is the dianion of 5,10,15,20-tetra\u00adkis\u00ad[2-\u00adphen\u00adyl]por\u00adph\u00adyrin} with an excess of KCN salts and an excess of the 18-crown-6 in chloro\u00adbenzene leads to the polymeric title compound catena-polyporphyrinato-1\u03baO5:2\u03ba4N,N\u2032,N\u2032\u2032,N\u2032\u2032\u2032:1\u2032\u03baO15}cobalt(III)potassium] dihydrate], {[CoK(CN)2(C12H24O6)(CN)2]\u2212 ion complex and one half of a . In the crystal, the polymeric chains and the lattice water mol\u00adecules are linked by N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds, as well as weak C\u2014H\u22efO, O\u2014H\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions into a three-dimensional supra\u00admolecular architecture.The reaction of Co DOI: 10.1107/S1600536814003596/xu5770Isup2.hklStructure factors: contains datablock(s) I. DOI: 987431CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit contains one half of the [Mg(TPP)(H2O)2] complex and one half of an 18-crown-6 mol\u00adecule. The average equatorial magnesium\u2013pyrrole N atom distance (Mg\u2014Np) is 2.071\u2005(1)\u2005\u00c5 and the axial Mg\u2014O(H2O) bond length is 2.213\u2005(1)\u2005\u00c5. The crystal packing is stabilized by two O\u2014H\u22efO hydrogen bonds between coordinating water mol\u00adecules and adjacent 18-crown-6 mol\u00adecules, and exhibits a one-dimensional supramolecular structure along the a axis. The supramolecular architecture is futher stabilized by weak C\u2014H\u22ef\u03c0 inter\u00adactions. The 18-crown-6 mol\u00adecule is disordered over two sets of sites with an occupancy ratio of 0.8:0.2.In the title compound, [Mg(C DOI: Click here for additional data file.10.1107/S1600536813001219/xu5669Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular network. \u03c0\u2013\u03c0 stacking inter\u00adactions between the imidazole rings [centroid\u2013centroid distances = 3.4914\u2005(15) and 3.6167\u2005(15)\u2005\u00c5] further stabilize the crystal structure. In the title compound, [Co(C DOI: 10.1107/S1600536812037579/hy2579Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CuI atom has a distorted tetra\u00adhedral coordination. The O\u2014Cu\u2014O angle is 80.07\u2005(8)\u00b0 and the P\u2014Cu\u2014P angle is 123.49\u2005(3)\u00b0. The crystal packing is stablized by intra\u00admolecular C\u2014H\u22efO inter\u00adactions and inter\u00admolecular C\u2014H\u22efO and O\u2014H\u22efO inter\u00adactions.In the title compound, [Cu(C DOI: 10.1107/S1600536811011470/fi2105Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ligator atoms comprise the two pyridine N atoms of the chelating di-2-pyridyl\u00adamine (dpa) ligand and two N atoms from two azide anions. The dpa ligand coordinates the Pd atom in a boat conformation, the dihedral angle between the pyridine rings being 24.4\u2005(1)\u00b0. The pyridine rings are somewhat inclined to the least-squares plane of the PdN4 unit, making dihedral angles of 29.02\u2005(9) and 26.47\u2005(9)\u00b0. The azide ligands are slightly bent, with N\u2014N\u2014N angles of 173.0\u2005(4) and 174.2\u2005(4)\u00b0. In the crystal, mol\u00adecules are connected by N\u2014H\u22efN and C\u2014H\u22efN hydrogen bonds, forming chains along the c axis. When viewed down the b axis, successive chains are stacked in opposite directions. Intra\u00admolecular C\u2014H\u22efN hydrogen bonds are also observed.In the title complex, [Pd(N II complexes [PdX2(dpa)] (X = Cl or Br), see: Rauterkus et al. 2(C10H9N3)] = 0.029wR(F2) = 0.073S = 1.062322 reflections181 parametersH-atom parameters constrainedmax = 0.71 e \u00c5\u22123\u0394\u03c1min = \u22120.41 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "These two constituents are linked by a remarkably short inter\u00adaction between the Br atom of the imidazolium cation [C\u2014Br = 1.85\u2005(3)\u2005\u00c5] and one N atom of the cyanidoargentate anion [Br\u22efN = 2.96\u2005(2)\u2005\u00c5], which is much less than the sum of the van der Waals radii (3.40\u2005\u00c5). The crystal studied was twinned by merohedry.The title structure, (C DOI: 10.1107/S1600536811051828/qk2020Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CoIII atom exhibits an octa\u00adhedral geometry, coordinated by four N atoms from the tris\u00ad(2-pyridyl\u00admeth\u00adyl)amine ligand with an average Co\u2014N distance of 1.953\u2005(2)\u2005\u00c5, and two cyanide C atoms with an average Co\u2014C distance of 1.895\u2005(2)\u2005\u00c5. The crystal packing is stabilized by inter\u00admolecular C\u2014H\u22efN and C\u2014H\u22efF inter\u00adactions.In the title complex, [Co(CN) DOI: 10.1107/S1600536811012001/vm2083Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the central Ti2(\u03bc2-O)2 fragment, the metal\u2013oxygen distances are significantly different [1.7962\u2005(19) and 1.9292\u2005(19)\u2005\u00c5]. In the crystal, the chloro\u00adform mol\u00adecule is anchored via an N\u2014H\u22efCl and a bifurcated C\u2014H\u22ef hydrogen bond. Slipped \u03c0\u2013\u03c0 stacking [shortest C\u22efC distance = 3.585\u2005(4)\u2005\u00c5] and C\u2014H\u22ef\u03c0 inter\u00adactions contribute to the coherence of the structure.In the title structure, [Ti DOI: 10.1107/S1600536813029656/qk2062Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536813029656/qk2062Isup3.molSupplementary material file. DOI: 969061 crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The two LaIII metal ions are linked by two bidentate bridging carboxyl\u00adate groups with a \u03ba2 O:O\u2032 coordination mode and two bidentate chelating bridging carboxyl\u00adate groups with a \u03ba3 O:O,O\u2032 coordination mode. The coordination sphere of lanthanum, completed by a terminal chloride and three water mol\u00adecules, adopts a distorted tricapped trigonal\u2013prismatic arrangement. N\u2014H\u22efCl, N\u2014H\u22efO and O\u2014Hwater\u22efCl hydrogen bonds, and slipped \u03c0\u2013\u03c0 inter\u00adactions between parallel benzene rings [centroid\u2013centroid distance of 3.647\u2005(3)\u2005\u00c5] are observed in the structure. These combine to stabilize a three-dimensional network.The tiltle complex, [La DOI: 10.1107/S1600536810052864/su2239Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Ru\u2014Ru distance and Ru\u2014O\u2014Ru angle are 3.2838\u2005(3)\u2005\u00c5 and 121.79\u2005(7)\u00b0, respectively, and the average Ru\u2014N(pyridine) bond length is 2.164\u2005(8)\u2005\u00c5. Several C\u2014H\u22efF, C\u2014H\u22efO and C\u2014H\u22efN inter\u00adactions generate a three-dimensional network in the crystal structure. \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.6389\u2005(3)\u2005\u00c5] between inversion-related 2,2\u2032-bipyridine rings are also observed.The hemerythrin-type dinuclear title complex, [Ru DOI: Click here for additional data file.10.1107/S1600536813003334/gg2109Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angles between the oxazole rings and their pendant rings are 2.0\u2005(3) and 24.3\u2005(2)\u00b0. The F atoms are disordered over two sites with occupancies of 0.627\u2005(3) and 0.373\u2005(3) in the phenyl\u00adene\u2013oxazol\u00adyl\u2013phenyl and in oxazol\u00adyl\u2013phenyl fragments, respectively. In the crystal structure, mol\u00adecules are linked through a network of C\u2014H\u22efF and weak \u03c0\u2013\u03c0 stacking inter\u00adactions.In the title compound, C DOI: 10.1107/S1600536810031235/fb2208Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The cation has a distorted octa\u00adhedral coordination environment and is surrounded by two N and two Cl atoms in the equatorial plane, while the coordinating water O atoms occupy the axial positions. The crystal exhibits nonmerohedral twinning with two domain states, the volume fractions of which were refined to 0.883\u2005(2) and 0.117\u2005(3). The crystal packing is stabilized by O\u2014H\u22efCl hydrogen-bond inter\u00adactions, forming two-dimensional networks lying parallel to (001). The crystal packing also features \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings, with centroid\u2013centroid separations of 3.493\u2005(3) and 3.545\u2005(3)\u2005\u00c5.The title mol\u00adecule, [CoCl For related structures, see: Li et al. 2(H2O)2] = 0.047wR(F2) = 0.149S = 1.162211 reflections88 parameters3 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.71 e \u00c5\u22123\u0394\u03c1min = \u22120.99 e \u00c5\u22123\u0394\u03c1CrysAlis CCD used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536813022484/fb2288Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In each mol\u00adecule, the four-coordinated SnIV atom exists in a distorted tetra\u00adhedral geometry and two intra\u00admolecular N\u2014H\u22efO hydrogen bonds with S(6) ring motifs are present. In one mol\u00adecule, the benzene ring of the 2-amino-3-nitro\u00adbenzoate ligand makes dihedral angles of 42.74\u2005(11), 89.66\u2005(13) and 53.04\u2005(10)\u00b0 with the three phenyl rings. The corresponding dihedral angles for the other mol\u00adecule are 6.29\u2005(11), 66.55\u2005(11) and 62.33\u2005(10)\u00b0. In the crystal, a weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction and a \u03c0\u2013\u03c0 stacking inter\u00adaction with a centroid\u2013centroid distance of 3.5877\u2005(12)\u2005\u00c5 are observed.The asymmetric unit of the title compound, [Sn(C DOI: 10.1107/S160053681101244X/is2696Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each PdII ion has a distorted square-planar coordination sphere, defined by three N atoms of the macrocyclic ligand and a chloride ion. The PdII complex cations and the methanol mol\u00adecules are linked through N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, forming a zigzag chain along [101]. An intra\u00admolecular N\u2014H\u22efCl hydrogen bond is also observed.In the crystal structure of the title compound, [Pd For dipters 2010; Goforth al. 2013. For pal al. 2013; Parker al. 1985. For a s al. 2003. For a s al. 2010.2(C36H42N6)Cl2](ClO4)2\u00b72C3H7NO\u00b72CH4O = 0.038wR(F2) = 0.097S = 1.085826 reflections324 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.80 e \u00c5\u22123\u0394\u03c1min = \u22120.81 e \u00c5\u22123\u0394\u03c1CrystalClear used to solve structure: SIR92 global, I. DOI: 10.1107/S1600536813022666/is5294Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The compound is a tetra\u00adnuclear ZnII complex centered about a fourfold roto-inversion axis, with the ligand coordinating in the doubly deprotonated form. The ZnII atom has a distorted square-pyramidal geometry being coordinated by one N and two O-atom donors from the doubly deprotonated L2\u2212 ligand, and by one N atom and one O-atom donor from a symmetry-related L2\u2212 ligand. In the crystal, four symmetry-related lattice water mol\u00adecules, centred about a fourfold roto-inversion axis, form a cyclic tetra\u00admer through O\u2014H\u22efO hydrogen bonds. These tetra\u00admers connect to the complex mol\u00adecules through O\u2014H\u22efN hydrogen bonds, forming a chain propagating along [100]. Neighbouring mol\u00adecules are linked by \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.660\u2005(2)\u2005\u00c5] involving the quinolidine rings.In the title compound, [Zn N-donor ligands, see: Palacios et al. 4]\u00b74H2O = 0.036wR(F2) = 0.087S = 1.032900 reflections217 parametersH-atom parameters constrainedmax = 0.27 e \u00c5\u22123\u0394\u03c1min = \u22120.23 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812018995/su2415Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "One RhII atom is five-coordinate, in a distorted pyramidal geometry, while the other is six-coord\u00adin\u00adate, with a disorted octa\u00adhedral geometry. For the six-coord\u00adinate RhII atom, the axial nitrile ligand shows a non-linear Rh\u2013nitrile coordination with an Rh\u2014N\u2014C bond angle of 166.4\u2005(4)\u00b0 and a nitrile N\u2014C bond length of 1.138\u2005(6)\u2005\u00c5. Each unique RhII atom is coordinated by a trans pair of N atoms and a trans pair of O atoms from the four acetamide ligands. The Neq\u2014Rh\u2014Rh\u2014Oeq torsion angles on the acetamide bridge varies between 12.55\u2005(11) and 14.04\u2005(8)\u00b0. In the crystal, the 3-methyl\u00adbenzo\u00adnitrile ring shows a \u03c0\u2013\u03c0 inter\u00adaction with an inversion-related equivalent [inter\u00adplanar spacing = 3.360\u2005(6)\u2005\u00c5]. A phenyl ring on one of the acetamide ligands also has a face-to-face \u03c0\u2013\u03c0 inter\u00adaction with an inversion-related equivalent [inter\u00adplanar spacing = 3.416\u2005(5)\u2005\u00c5].In the title compound, [Rh DOI: 10.1107/S1600536813029838/pk2497Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Am. Mineral.54, 19\u201330], all non-H atoms were refined with anisotropic displacement parameters and H-atoms were located by difference Fourier methods and refined from X-ray diffraction data. The structure is built up from nearly regular [Al(H2O)6]3+ octa\u00adhedra and infinite double-stranded chains [Na(SO4)2]3\u2212 that extend parallel to [001]. The Na+ cation has a strongly distorted octa\u00adhedral coordination by sulfate O atoms [Na\u2014O = 2.2709\u2005(11) \u2013 2.5117\u2005(12)\u2005\u00c5], of which five are furnished by the chain-building sulfate group S2O4 and one by the non-bridging sulfate group S1O4. The [Na(SO4)2]3\u2212 chain features an unusual centrosymmetric group formed by two NaO6 octa\u00adhedra and two S2O4 tetra\u00adhedra sharing five adjacent edges, one between two NaO6 octa\u00adhedra and two each between the resulting double octa\u00adhedron and two S2O4 tetra\u00adhedra. These groups are then linked into a double-stranded chain via corner-sharing between NaO6 octa\u00adhedra and S2O4 tetra\u00adhedra. The S1O4 group, attached to Na in the terminal position, completes the chains. The [Al(H2O)6]3+ octa\u00adhedron (\u2329Al\u2014O\u232a = 1.885\u2005(11)\u2005\u00c5) donates 12 comparatively strong hydrogen bonds (O\u22efO = 2.6665\u2005(14) \u2013 2.7971\u2005(15)\u2005\u00c5) to the sulfate O atoms of three neighbouring [Na(SO4)2]3\u2212 chains, helping to connect them in three dimensions, but with a prevalence parallel to (010), the cleavage plane of the mineral. Compared with the previous work on tamarugite, the bond precision of Al\u2014O bond lengths as an example improved from 0.024 to 0.001\u2005\u00c5.The crystal structure of tamarugite [sodium aluminium bis\u00ad(sulfate) hexa\u00adhydrate] was redetermined from a single crystal from Mina Alcaparossa, near Cerritos Bayos, southwest of Calama, Chile. In contrast to the previous work [Robinson & Fang 1969. Am. Min DOI: 10.1107/S1600536813025154/pj2005Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The crystal structure is stabilized by weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0(arene) inter\u00adactions. The Ru\u2014\u03b75-cyclopentadienyl centroid bond length is 1.946\u2005(11)\u2005\u00c5In the title mol\u00adecule, [Ru(C DOI: 10.1107/S1600536810033180/lh5113Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The metal atom is further chelated by a carboxyl\u00adate group and is covalently bonded to a monodentate carboxyl\u00adate group as well as to a monodentate sulfonate group in a distorted square anti\u00adprismatic geometry. The coordinating water molecules and the lattice water molecules, one of which is disordered over two positions [major component 65\u2005(3)%], are hydrogen bonded to the network.The 4-sulfophthalate trianion in the polymeric complex, {[Er(C III derivative, see: Xiao et al. Eu al. 2010.8H3O7S)(C12H8N2)(H2O)2]\u00b72H2O = 0.037wR(F2) = 0.095S = 1.094014 reflections353 parameters33 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.97 e \u00c5\u22123\u0394\u03c1min = \u22121.21 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812003467/xu5455Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atom in the title compound, {[Pb2(C7H6NO2)4]}n, is chelated by two 3-aminobenzoato ligands in a distorted pentagonal-bipyramidal coordination geometry with five oxygen donors in the equatorial positions, one nitro\u00adgen donor and one oxygen donor in the axial positions. Two mol\u00adecules are linked through a centre of inversion, forming a dinuclear entity. These entities are linked in a \u03bc3-bridging mode through the amino N atom and two carboxyl\u00adate O atoms into a chain along the b axis. Classical inter\u00admolecular N\u2014H\u22efO hydrogen bonding is observed in the structure. The supra\u00admolecular structure is consolidated by \u03c0\u2013\u03c0 stacking inter\u00adactions with centroid\u2013centroid distances between benzene rings of 3.837\u2005(8)\u2005\u00c5.The Pb DOI: 10.1107/S1600536810041322/rk2229Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the trigonal\u2013pyramidal Ru3P core, one RuII atom is coordinated by a triphenyl\u00adphosphane ligand in a terminal fashion. Two hydride ligands bridge over two Ru\u2014Ru bonds. These Ru\u2014Ru bonds [2.9400\u2005(4) and 2.9432\u2005(4)\u2005\u00c5] are slightly longer than the nonhydride-bridged Ru\u2014Ru bond [2.8146\u2005(4)\u2005\u00c5]. The terminal triphenyl\u00adphosphane ligand coordinates to the RuII atom, which is involved in two hydride bridges.In the crystal structure of the title compound, [Ru H2(C18H15P)(CO)8] = 0.030wR(F2) = 0.118S = 1.238304 reflections444 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.74 e \u00c5\u22123\u0394\u03c1min = \u22121.79 e \u00c5\u22123\u0394\u03c1PROCESS-AUTO I, global. DOI: 10.1107/S1600536812039499/nc2293Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The carbonyl and chloride ligands in the cation adopt a mutually cis arrangement and these are disordered over two sets of sites with site occupancies of 0.721\u2005(6) and 0.279\u2005(6). The Ru\u2014N bond length [2.117\u2005(2)\u2005\u00c5] trans to the carbonyl ligand is slightly longer than the average of the other Ru\u2014N bond lengths (2.08\u2005\u00c5), which can be explained by the expected trans influence of the carbonyl group. In the crystal, weak C\u2014H\u22efF inter\u00adactions are observed between the complex cation and the PF6\u2212 anion, leading to the formation of a three-dimensional supramolecular structure. The crystal studied was an inversion twin with twin fractions of 0.78\u2005(4) and 0.22\u2005(4).In the title compound, [RuCl(C Cl]+, see: Ishida et al. 2(CO)]\u00b7PF6 = 0.032wR(F2) = 0.062S = 1.085177 reflections330 parametersH-atom parameters constrainedmax = 1.37 e \u00c5\u22123\u0394\u03c1min = \u22121.28 e \u00c5\u22123\u0394\u03c1Absolute structure: Flack 1983, 2249 FrFlack parameter: 0.22 (4)CrystalClear-SM used to solve structure: SIR97 global, I. DOI: Click here for additional data file.10.1107/S1600536812048246/is5220Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The mol\u00adecular structure is stabilized by weak C\u2014H\u22efO and C\u2014H\u22efF hydrogen-bond inter\u00adactions. The crystal structure is stabilized by \u03c0\u2013\u03c0 stacking inter\u00adactions (centroid\u2013centroid distance = 3.951\u2005\u00c5).The title compound, [Ir(C DOI: 10.1107/S1600536813026160/bx2449Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, pairs of mol\u00adecules form inversion dimers via N\u2014H\u22efCl hydrogen bonds. In addition, intra\u00admolecular N\u2014H\u22efCl and weak C\u2014H\u22efCl, C\u2014H\u22efN, N\u2014H\u22ef\u03c0 and C\u2014H\u22ef\u03c0 hydrogen bonds are observed. One of the Cl atoms of the solvent mol\u00adecule is disordered over two sites with refined occupancies of 0.62\u2005(1) and 0.38\u2005(1).In the title compound, [RuCl DOI: 10.1107/S1600536812014080/lh5442Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Within the pcp ligands, the dihedral angles between the polycyclic skeletons and pendant pyridine rings are 6.2\u2005(2) and 8.3\u2005(2)\u00b0. In the crystal, mol\u00adecules are linked by O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds. Several aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions [shortest centroid\u2013centroid separation = 3.516\u2005(3)\u2005\u00c5] are also observed.In the title compound, [Mn(C DOI: 10.1107/S1600536811004569/hb5800Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The NiII atom is in a distorted octa\u00adhedral coordination by two N atoms in a cis disposition [Ni\u2014N = 2.0753\u2005(16) and 2.1048\u2005(16)\u2005\u00c5] and by four water O atoms [Ni\u2014O = 2.0500\u2005(15)\u20132.0822\u2005(15)\u2005\u00c5]. The crystal structure is completed by four further non-coordinating water mol\u00adecules and all constituents are linked in a three-dimensional manner by an extensive system of 16 O\u2014H\u22efO hydrogen bonds.The title structure, [Ni(C DOI: 10.1107/S1600536812027122/qk2036Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The 4-(dimethyl\u00adamino)\u00adpyridinium cations are protonated at the pyridine N atoms and form N\u2014H\u22efCl hydrogen bonds with Cl\u2212 counter-ions. The dimethyl\u00adamino groups (C/N/C) lie close to the plane of the pyridinium rings, making dihedral angles of 1.6\u2005(6)\u00b0 and 6.5\u2005(3)\u00b0. In the crystal, the [Nd2Cl4(H2O)10]2+ dications are linked via O\u2014H\u22efO and O\u2014H\u22efCl hydrogen bonds, forming sheets lying parallel to the bc plane. These sheets are linked via O\u2014H\u22efCl hydrogen bonds, forming a three-dimensional network. The 4-(dimethyl\u00adamino)\u00adpyridinium cations are located in the cavities and linked to the framework by C\u2014H\u22efCl interactions.The title compound, (C DOI: Click here for additional data file.10.1107/S1600536812041724/su2506Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II complex, [Cu(C7H3NO4)(C6H5NO2)(H2O)]\u00b7H2O, the environment of the Cu2+ ion is a distorted square pyramid with the axial site occupied by the O atom from the coordinated water mol\u00adecule and the square base formed by two O and two N atoms from the tridentate anion and the neutral monodentate pyridine-3-carboxylic acid ligand. O\u2014H\u22efO hydrogen bonds, as well as \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.945\u2005(3)\u2005\u00c5] contribute to the stabilization of this structure.In the title Cu DOI: 10.1107/S160053681101926X/yk2006Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular C\u2014H\u22efBr hydrogen bonds and \u03c0\u2013\u03c0 stacking between the pyridine rings in the bc plane [centroid\u2013centroid distance = 3.725\u2005(3)\u2005\u00c5] are present in the crystal structure.In the title compound, [CoBr DOI: Click here for additional data file.10.1107/S1600536812041980/xu5630Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The crystal packing exhibits a short O\u22efBr inter\u00adaction [Br\u22efO = 3.185\u2005(3)\u2005\u00c5] and a weak inter\u00admolecular C\u2014H\u22efO contact.The title compound, C DOI: 10.1107/S1600536811010051/fy2003Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The octa\u00adhedral environment around niobium is slightly distorted with Nb\u2014O distances in the range 1.8603\u2005(15)\u20132.1083\u2005(15)\u2005\u00c5 and an Nb\u2014Cl distance of 2.4693\u2005(9)\u2005\u00c5. The O\u2014Nb\u2014O angles vary between 80.74\u2005(6) and 100.82\u2005(7)\u00b0, while the trans Cl\u2014Nb\u2014O angle is 167.60\u2005(5)\u00b0. There are no hydrogen bonds observed, only an inter\u00admolecular C\u2014H\u22efO inter\u00adaction.In the title compound, [Nb(CH DOI: 10.1107/S1600536810021719/pv2289Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the components are linked into a three-dimensional framework by inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and N\u2014H\u22efN and weak C\u2014H\u22efO hydrogen bonds. In addition, \u03c0\u2013\u03c0 stacking inter\u00adactions with centroid\u2013centroid distances in the range 3.4809\u2005(7)\u20133.8145\u2005(6)\u2005\u00c5 are observed.The asymmetric unit of the title complex, (C DOI: 10.1107/S1600536811027917/lh5282Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds connect mol\u00adecules into chains along [100] and there are \u03c0\u2013\u03c0 stacking inter\u00adactions between pairs of chains with a centroid\u2013centroid distance of 3.5485\u2005(15)\u2005\u00c5.In the title compound, C DOI: 10.1107/S160053681002979X/lh5093Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CdII atom is seven-coordinated in a distorted penta\u00adgonal\u2013bipyramidal configuration, defined by five O atoms from the carboxyl\u00adate groups of three 7-oxabicyclo\u00ad[2.2.1]heptane-2,3-dicarboxyl\u00adato ligands and two N atoms from the 2,2\u2032-bipyridine ligand. Two O atoms link two CdII atoms, forming a dinuclear center: the Cd\u2014O\u2014Cd bridging angle is 110.19\u2005(6)\u00b0. The polymeric structure extends along [100] and is linked by inter\u00admolecular O\u2014H\u22efO hydrogen bonds involving the solvent water molecule. Extensive \u03c0\u2013\u03c0 stacking exists between 2,2-bypiridine ligands along [010] with centroid-centroid distance of 3.650\u2005(2)\u2005\u00c5The title compound, {[Cd(C DOI: 10.1107/S1600536811036634/zb2016Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the components are linked via inter\u00admolecular O\u2014H\u22efO hydrogen bonds, forming layers parallel to (001). Additional stabilization within these layers is provided by \u03c0\u2013\u03c0 [centroid\u2013centroid distances of 3.7848\u2005(9)\u20134.4231\u2005(9)\u2005\u00c5] stacking inter\u00adactions.The central CuN DOI: 10.1107/S1600536811013808/bq2292Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The range of Ag\u2014O bond lengths is 2.169\u2005(2)\u20132.433\u2005(2)\u2005\u00c5, whereas the Ag\u22efAg separations are in the range 2.8674\u2005(4)\u20133.6256\u2005(5)\u2005\u00c5. The 2-methyl\u00adbenzoate groups are oriented at a dihedral angle of 60.7\u2005(1)\u00b0 with respect to each other.The crystal structure of the title compound, [Ag DOI: 10.1107/S1600536811016801/ez2241Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the dinuclear complex mol\u00adecules are linked into one-dimensional supra\u00admolecular columns parallel to the b axis by O\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.7259\u2005(11)\u2005\u00c5].The title complex, [Cu DOI: 10.1107/S1600536812039839/rz5004Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Intra\u00admolecular O\u2014H\u22efN hydrogen bonds between the hy\u00addroxy O atoms and the imine N atoms, with O\u22efN distances in the range 2.607\u2005(3)\u20132.665\u2005(3)\u2005\u00c5, form nearly planar six-membered rings. In the crystal, weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds occur and several intra- and inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions are present between adjacent benzene rings, with a shortest centroid\u2013centroid distance of 3.507\u2005(2)\u2005\u00c5.In the title compound, C DOI: 10.1107/S1600536810047185/is2631Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The HgII atom is four-coordinated in a distorted tetra\u00adhedral geometry by one terminal Cl atom, two bridging Cl atoms, and one C atom from the ylidic ligand, resulting in a polymeric chain parallel to [010]. The terminal Cl atom is more strongly bound to the HgII ion [2.3916\u2005(9)\u2005\u00c5] than the bridging Cl atoms. The bridge is asymmetric, as indicated by the two different Hg\u2014Cl(bridging) bond lengths [2.5840\u2005(8) and 2.7876\u2005(8)\u2005\u00c5]. Intra\u00admolecular C\u2014H\u22efO and weak C\u2014H\u22efCl contacts stabilize the polymeric chain. In the crystal, adjacent chains inter\u00adact via C\u2014H\u22efO hydrogen bonds. In the title organometallic polymer, [HgCl II complex, see: Ebrahim et al. ] = 0.025wR(F2) = 0.067S = 1.036143 reflections307 parametersH-atom parameters constrainedmax = 0.96 e \u00c5\u22123\u0394\u03c1min = \u22120.45 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536812043073/bh2457Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II title compound, [Cd2(C9H10NO2)4(C6H6N2O)2(H2O)2], each seven-coordin\u00adated CdII atom is chelated by the carboxyl\u00adate groups of the two 4-(dimethyl\u00adamino)\u00adbenzoate (DMAB) anions; the two monomeric units are bridged through the two O atoms of the two carboxyl groups. In the crystal structure, inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network. \u03c0\u2013\u03c0 contacts between the pyridine rings [centroid\u2013centroid distance = 3.974\u2005(1)\u2005\u00c5] may further stabilize the structure. Weak C\u2014H\u22ef\u03c0 inter\u00adactions are also observed.In the centrosymmetric dimeric Cd DOI: 10.1107/S160053681002163X/xu2771Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Zn\u2014Br bond lengths range from 2.3901\u2005(19) to 2.449\u2005(2)\u2005\u00c5 and the Br\u2014Zn\u2014Br angles range from 107.09\u2005(8) to 112.48\u2005(8)\u00b0. In the crystal, each [ZnBr4]2\u2212 anion is connected to four cations through two N\u2014H\u22efBr and two C\u2014H\u22efBr hydrogen bonds, forming two-dimensional \u22ef(cation)2\u22efanion\u22ef(cation2)\u22ef sheets parallel to the bc plane. Within each sheet, the anions are arranged in stacks with no significant inter-anion Br\u22efBr inter\u00adactions [the shortest being > 4.3\u2005\u00c5], while the cations are in chains, with weak \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.991\u2005\u00c5] between cations inter\u00adacting with the same anion.In the title compound, (C DOI: Click here for additional data file.10.1107/S1600536812040925/pv2593Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The uncoordinating phen mol\u00adecule is approximately parallel to the 1,10-phenanthrolin-1-ium (Hphen) anion [dihedral angle = 3.3\u2005(4)\u00b0]. The centroid\u2013centroid distance of 3.801\u2005(5)\u2005\u00c5 between pyridine rings suggests the existence of \u03c0\u2013\u03c0 stacking. The crystal structure contains an extensive network of classical O\u2014H\u22efO and N\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds. C\u2014H\u22ef\u03c0 inter\u00adactions between phen and 4-hy\u00addroxy\u00adbenzoate is also present in the crystal structure. In the crystal, the uncoordinating phen is equally disordered over two sites about an inversion center.In the title compound, C DOI: 10.1107/S1600536810051767/xu5103Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angle between the benzimidazole and benzene rings is 9.57\u2005(1)\u00b0. In the crystal, mol\u00adecules are linked by weak N\u2014H\u22efCl hydrogen bonds into layers parallel to the bc plane. \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances in the range 3.4452\u2005(8)\u20133.8074\u2005(8)\u2005\u00c5 are also observed.In the title benzimidazole mononuclear complex, [ZnCl DOI: 10.1107/S1600536811026572/rz2622Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The NiII ion is coordinated in a distorted square-planar geometry by the tetra\u00addentate ligand. The dihedral angle between the two symmetry-related benzene rings is 47.12\u2005(8)\u00b0. In the crystal, pairs of symmetry-related O\u2014H\u22efO hydrogen bonds form R 2 2(6) ring motifs. In addition, there are weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds, and \u03c0\u2013\u03c0 stacking inter\u00adactions with a centroid\u2013centroid distance of 3.4760\u2005(8)\u2005\u00c5.In the title compound, [Ni(C DOI: 10.1107/S1600536811054262/lh5396Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The L\u2212 ligands link the CdII atoms into a plane parallel to (100), with one ligand being tridentate, coordinating via the N atom and chelating a second Cd atom, and the other being bidentate, bridging two Cd atoms via the N and one O atom.. This two-dimensional network extends into a double-layer network by \u03c0\u2013\u03c0 inter\u00adactions, with centroid\u2013centroid distances of 3.680\u2005(2) and 3.752\u2005(2)\u2005\u00c5. Another type of \u03c0\u2013\u03c0 inter\u00adaction between pyridine rings [centroid\u2013centroid distance = 3.527\u2005(2)\u2005\u00c5] leads to a three-dimensional supra\u00admolecular architecture.In the title compound, [Cd(C DOI: 10.1107/S1600536812001031/vn2029Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atom in the title compound, [Cd(C5H10NS2)2(C10H8N2)], exists in an N2S4 donor set defined by two chelating dithio\u00adcarbamate anions as well as a 2,2\u2032-bipyridine ligand. The coordination geometry approximates a trigonal prism. The crystal packing features weak C\u2014H\u22efS inter\u00adactions, leading to linear supra\u00admolecular chains along the a axis. The primary connections between these are by \u03c0\u2013\u03c0 stacking inter\u00adactions [ring centroid distance between centrosymmetrically related pyridyl rings = 3.7455\u2005(10)\u2005\u00c5]. Overall, the crystal structure may be described as comprising double layers of mol\u00adecules that stack along the b axis.The Cd DOI: 10.1107/S1600536811012414/hb5834Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The calcium ion is six-coordinated and is bonded to all phen\u00adoxy O atoms from both zwitterionic saltren mol\u00adecules. There are strong intra\u00admolecular N\u2014H\u22efO hydrogen bonds. The cations are linked into chains via weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.306\u2005(3) and 3.415\u2005(3)\u2005\u00c5].The title complex, [Ca(C DOI: 10.1107/S1600536810053961/pv2371Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "An intra\u00admolecular O\u2014H\u22efO hydrogen bond occurs. In the crystal, inter\u00admolecular O\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network. \u03c0\u2013\u03c0 inter\u00adactions between the imidazole and benzene rings [centroid\u2013centroid distance = 3.9031\u2005(17)\u2005\u00c5] consolidate the crystal packing.In the title compound, [Co(C DOI: 10.1107/S1600536811029059/hy2449Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each PtII ion is four-coordinated in an essentially square-planar environment by four pyridine N atoms derived from the two chelating di-2-pyridyl\u00adamine (dpa) ligands, and the PtN4 unit is exactly planar. The chelate ring formed by the dpa ligand displays a boat conformation, with dihedral angles between the pyridine rings of 35.9\u2005(2) and 41.0\u2005(2)\u00b0. The complex cations, Br\u2212 anions and solvent water mol\u00adecules are linked by O\u2014H\u22efBr, N\u2014H\u22efBr, C\u2014H\u22efO and C\u2014H\u22efBr hydrogen bonds, forming a three-dimensional network.The asymmetric unit of the title compound, [Pt(C II and PtII complexes, see: \u017divkovi\u0107 et al. 2]Br2\u00b7H2O = 0.029wR(F2) = 0.069S = 1.054109 reflections274 parametersH-atom parameters constrainedmax = 1.43 e \u00c5\u22123\u0394\u03c1min = \u22121.21 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CdII ion is coordinated in a disorted square-pyramidal CdBr2N2S environment with one of the Br atoms in the apical site. In the crystal structure, the benzimidazole ring systems are involved in weak inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.606\u2005(2) and 3.753\u2005(2)\u2005\u00c5]. Further stabilization is provided by weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds. The methyl H atoms of the dimethyl\u00adformamide solvent mol\u00adecule are disordered about a mirror plane.In the title compound, [CdBr DOI: 10.1107/S1600536810028448/lh5084Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atom in the title compound, [Zn(C2H3O2)2(C14H13N3)], is coordinated by an N2O3 donor set defined by the quinolinyl- and pyrazolyl-N atoms of the chelating heterocyclic ligand, and three carboxyl\u00adate-O atoms derived from the monodentate and bidentate carboxyl\u00adate ligands. Distortions from the ideal square-pyramidal coordination geometry relate to the restricted bite angle of the chelating ligands, i.e. O\u2014Zn\u2014O = 59.65\u2005(5) and N\u2014Zn\u2014N = 76.50\u2005(6)\u00b0, and the close approach of the non-coordinating carbonyl atom [Zn\u22efO = 2.858\u2005(2)\u2005\u00c5]. In the crystal, mol\u00adecules are consolidated into a three-dimensional architecture by C\u2014H\u22efO inter\u00adactionsThe Zn DOI: 10.1107/S1600536812025664/hb6839Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Both Na atoms exhibit a distorted octa\u00adhedral coordination geometry and are coordinated by five water O atoms and the terminal O atom from a 7-hy\u00addroxy\u00adcoumarin ligand. Four of the water mol\u00adecules are bridging, whereas the fifth is terminal. Na\u2014O bond distances are in the range 2.288\u2005(2)\u20132.539\u2005(2)\u2005\u00c5. In the chains, extending parallel to [100], adjacent Na atoms are separated by 3.60613\u2005(7)\u2005\u00c5. The uncoordinated water mol\u00adecules and 7-hy\u00addroxy\u00adcoumarin phenolate anions are located between the chains and are hydrogen bonded to the chains.The asymmetric unit of the title compound, {[Na(C HOC), see: Toyama et al. (H2O)3](C9H5O3)\u00b7H2O = 0.056 wR(F 2) = 0.153 S = 1.13 7251 reflections573 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.32 e \u00c5\u22123 \u0394\u03c1min = \u22120.31 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008SHELXTL (Sheldrick, 2008SHELXTL.Data collection: 10.1107/S160053681002341X/rk2209sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S160053681002341X/rk2209Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Fe\u2014N bond length is 1.924\u2005(3)\u2005\u00c5 and the Fe\u2014Cp* centroid distance is 1.722\u2005\u00c5.In the structure of the title compound, [Fe{\u03b7 DOI: 10.1107/S1600536811021350/hg5040Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The hy\u00addroxy group of the hy\u00addroxy\u00adacetate ligand links with the counter NO3 \u2212 anion via a pair of bifurcated O\u2014H\u22efO hydrogen bonds. The centroid\u2013centroid distance of 3.5676\u2005(14)\u2005\u00c5 between benzene rings of parallel phen ligands of adjacent mol\u00adecules suggests the existence of \u03c0\u2013\u03c0 stacking. Weak inter\u00admolecular C\u2014H\u22efO hydrogen bonding is also present in the crystal structure.In the title compound, [Cu(C DOI: 10.1107/S1600536811013110/xu5175Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The NiII atom is located on a twofold rotation axis and is bonded to four N atoms at distances of 2.037\u2005(8) and 2.109\u2005(9)\u2005\u00c5, and to two S atoms at a distance of 2.406\u2005(3)\u2005\u00c5, leading to a distorted octa\u00adhedral coordination. The angle between the mean planes of the coordinating moieties of the two symmetry-related tridentate ligands is 83.3\u2005(2)\u00b0. In the crystal, complex mol\u00adecules are linked by weak C\u2014H\u22efS hydrogen bonds, \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings [centroid\u2013centroid distance = 3.775\u2005(9)\u2005\u00c5] and C\u2014H\u22ef\u03c0 inter\u00adactions. The hydrogen-bonding inter\u00adactions lead to the formation of layers parallel to (010); \u03c0\u2013\u03c0 inter\u00adactions link these layers into a three-dimensional network.The title compound, [Ni(C DOI: Click here for additional data file.10.1107/S1600536813013032/wm2742Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536813013032/wm2742Isup3.molSupplementary material file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The pyridyl N atom of the pydc2\u2212 anion and one pyridyl N atom of bpy occupy the axial positions. O\u2014H\u22efO hydrogen bonds involving the ethanol solvent mol\u00adecule as donor and a carboxyl\u00adate O atom as acceptor atoms, as well as C\u2014H\u22efO hydrogen bonds, together with \u03c0\u2013\u03c0 stacking inter\u00adactions between adjacent aromatic rings (average centroid\u2013centroid distance = 3.577\u2005\u00c5), seem to be effective in the stabilization of the crystal packing, resulting in the formation of a three-dimensional structure.In the title compound, [V(C O(C10H8N2)]\u00b7C2H6O = 0.058 wR(F 2) = 0.144 S = 1.03 2959 reflections263 parameters18 restraintsH-atom parameters constrainedmax = 0.40 e \u00c5\u22123 \u0394\u03c1min = \u22120.47 e \u00c5\u22123 \u0394\u03c1 X-AREA used to solve structure: SHELXTL (Sheldrick, 2008SHELXTL; molecular graphics: SHELXTL; software used to prepare material for publication: SHELXTL and PLATON (Spek, 2009Data collection: 10.1107/S1600536811002376/wm2448sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536811002376/wm2448Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The quaternary C atoms of the complexed arene ring are located at the longest distance from the Fe atom, with Fe\u2014C distances of 2.112\u2005(4) and 2.105\u2005(3)\u2005\u00c5, which are slightly longer than the average Fe\u2014C distance for this ring (2.083\u2005\u00c5). The Fe ion is located 1.660\u2005(1) and 1.543\u2005(1)\u2005\u00c5, respectively, from the cyclo\u00adpenta\u00addienyl and the complexed arene ring.At 296\u2005(2)\u2005K, both complexed rings in the iron(II) complex cation of the title salt, [Fe(C DOI: 10.1107/S1600536810033179/si2288Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The 2-phenyl\u00adpyridine ligands are not planar, the dihedral angles between the pyridine and benzene rings being 50.1\u2005(2) and 45.7\u2005(2)\u00b0. An inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adaction between the six-membered rings is present, the ring centroid\u2013centroid distance being 3.898\u2005(4)\u2005\u00c5.In the title complex, [PdI DOI: 10.1107/S1600536811055425/is5038Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The central CuII atom exhibits a distorted square-planar coordination geometry, defined by two O atoms, one N atom from the ligand and one pyridine N atom with Cu\u2014N distances of 1.874\u2005(4) and 1.963\u2005(4)\u2005\u00c5, while the Cu\u2014O distances are 1.857\u2005(3) and 1.890\u2005(3)\u2005\u00c5. An intra\u00admolecular O\u2014H\u22efN inter\u00adaction occurs.The mononuclear title complex, [Cu(C DOI: 10.1107/S1600536810047719/bx2325Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII atom shows a distorted square-planar geometry. The dihedral angle between the two aromatic rings is 48.16\u2005(13)\u00b0. The crystal structure is stabilized by inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances in the range 3.485\u2005(2)\u20133.845\u2005(3)\u2005\u00c5.The asymmetric unit of the title compound, [Cu(C DOI: 10.1107/S1600536810053183/jh2247Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Of the two independent dimethyl\u00adformamide mol\u00adecules, one is hydrogen bonded to a single \u2013COOH group, while one links two adjacent \u2013COOH groups via a strong accepted O\u2014H\u22efO and a weak donated C(O)\u2014H\u22efO hydrogen bond. Two of these last mol\u00adecules and the two \u2013COOH groups form a centrosymmetric hydrogen-bonded ring in which the CH=O and the \u2013COOH groups by disorder adopt two alternate orientations in a 0.44:0.56 ratio. These hydrogen bonds link the CuII complex mol\u00adecules and the dimethyl\u00adformamide solvent mol\u00adecules into infinite chains along [-111]. Slipped \u03c0\u2013\u03c0 stacking inter\u00adactions between two centrosymmetric pyridine rings (centroid\u2013centroid distance = 3.63\u2005\u00c5) contribute to the coherence of the structure along [0-11].In the title compound, [Cu DOI: Click here for additional data file.10.1107/S1600536812051422/qk2049Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Four water O atoms in the equatorial plane around the NiII ion [Ni\u2014O = 2.052\u2005(2) and 2.079\u2005(2)\u2005\u00c5] form a slightly distorted square-planar arrangement, which is completed up to a distorted octa\u00adhedron by the two N atoms [Ni\u2014N = 2.075\u2005(3)\u2005\u00c5] from two isonicotinamide ligands. In the anion, the carboxyl\u00adate group is twisted from the attached benzene ring by 8.8\u2005(3)\u00b0. In the crystal, a three-dimensional hydrogen-bonding network, formed by classical O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, consolidates the crystal packing, which also exhibits \u03c0\u2013\u03c0 inter\u00adactions between the benzene and pyridine rings, with centroid\u2013centroid distances of 3.455\u2005(2) and 3.621\u2005(2)\u2005\u00c5, respectively.The asymmetric unit of the title compound, [Ni(C DOI: 10.1107/S1600536812002218/cv5237Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Neighbouring units inter\u00adact through \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.380\u2005(3) and 3.283\u2005(4)\u2005\u00c5]. Finally, three types of O\u2014H\u22efO hydrogen bonds exist between coordinated dissociative water mol\u00adecules and hybridization water mol\u00adecules and carboxyl\u00adate O atoms, resulting in a two-dimensional network parallel to (010).In the title compound, [Cd(C DOI: 10.1107/S1600536810045551/nk2061Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordination environments around the MoVI atoms are distorted octa\u00adhedral, defined by two oxide ligands and an N2O2 donor set of the tetra\u00addentate Schiff base in each mol\u00adecule. The dihedral angles between the benzene rings in the mol\u00adecules are 76.2\u2005(3) and 77.7\u2005(3)\u00b0. An inter\u00adesting feature of the crystal structure is the presence of Br\u22efBr contacts , which are shorter than the sum of the van der Waals radius of Br atoms (3.70\u2005\u00c5). The crystal structure is further stabilized by inter\u00admolcular C\u2014H\u22efBr and C\u2014H\u22ef\u03c0 inter\u00adactions. The crystal under investigation was twinned by nonmerohedry in a 0.053\u2005(1):0.947\u2005(1) ratio.The asymmetric unit of the title compound, [Mo(C O2] = 0.048wR(F2) = 0.101S = 1.0111292 reflections546 parametersH-atom parameters constrainedmax = 1.28 e \u00c5\u22123\u0394\u03c1min = \u22121.11 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXTL global, I. DOI: 10.1107/S1600536812039785/wm2668Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The inter\u00adnal cohesion of the mol\u00adecule is enhanced by an intra\u00admolecular O\u2014H\u22efO hydrogen bond. Inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 contacts [centroid\u2013centroid distance = 3.6549\u2005(2)\u2005\u00c5] define two-dimensional networks parallel to (001), which are further connected by weaker C\u2014H\u22efO inter\u00adactions into a weakly connected three-dimensional supra\u00admolecular framework.In the centrosymmetric title compound, [Cd(C DOI: 10.1107/S1600536812033739/bg2473Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings of the pydc ligands [centroid\u2013centroid distance = 3.4714\u2005(14)\u2005\u00c5] are present. C=O\u22ef\u03c0 inter\u00adactions between the carbonyl groups and pyridine rings [O\u22efcentroid distances = 3.150\u2005(2) and 3.2233\u2005(19)\u2005\u00c5] are also observed.In the title compound, (C DOI: 10.1107/S1600536811024378/hy2413Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.5469\u2005(16)\u2005\u00c5] between the pyridyl and benzene rings. The shortest Ag\u22efAg distance is 3.2574\u2005(5)\u2005\u00c5.In the title compound, [Ag(C DOI: 10.1107/S1600536810025511/zl2286Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "They are inclined to one another at an angle of 1.95\u2005(2)\u00b0 and lie 3.309\u2005(2)\u00c5 away from each other. The ethyl\u00adidene\u00adthio\u00adcarbonohydrazide fragment is planar, with an r.m.s. deviation of 0.0347\u2005(2)\u2005\u00c5 from the mean plane of its eight non-H atoms, and makes dihedral angles of 21.78\u2005(1) and 19.97\u2005(1)\u00b0 with respect to the two Cp rings. The mol\u00adecule adopts a trans geometry about the C=N double bond. In the crystal, N\u2014H\u22ef(N/S) and C\u2014H\u22efS inter\u00adactions stack the mol\u00adecules in an inverse fashion along the b axis.In the title compound, [Fe(C DOI: Click here for additional data file.10.1107/S1600536812044078/sj5273Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In each mol\u00adecule, the SnIV atom is coordinated by O, N and S atoms from a \u00adthio\u00adsemicarbazonato ligand and two C atoms from phenyl rings in a distorted trigonal\u2013bipyramidal geometry. Weak inter\u00admolecular N\u2014H\u22efO and N\u2014H\u22efS hydrogen bonds link four mol\u00adecules into a centrosymmetric tetra\u00admer. The crystal packing exhibits short inter\u00admolecular S\u22efS contacts of 3.335\u2005(3)\u2005\u00c5.The asymmetric unit of the title compound, [Sn(C DOI: 10.1107/S1600536810054024/cv5008Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Cu atoms are bridged by two O atoms of the monodentate phenyl\u00adacetate groups [Cu\u2014O = 1.9808\u2005(14) and 2.3668\u2005(14)\u2005\u00c5]. The longer of the two bridging Cu\u2014O bonds takes the apical position of the distorted square-pyramidal environment of the Cu atom, which is completed by two N atoms of the chelate 2,2\u2032-bipyridine ligand [Cu\u2014N = 2.0107\u2005(17) and 2.0234\u2005(16)\u2005\u00c5]. The mol\u00adecules are assembled into stacks along [100] through \u03c0\u2013\u03c0 inter\u00adactions with inter\u00adplanar distances of 3.630\u2005(4) and 3.407\u2005(3)\u2005\u00c5; the resulting stacks are further connected into a three-dimensional supra\u00admolecular architecture by O\u2014H\u22efO and C\u2014H\u22efO hydrogen-bonding inter\u00adactions.The mol\u00adecule of the binuclear title complex, [Cu DOI: 10.1107/S1600536811035483/ya2144Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Cu(4-methyl\u00adpyridine)4 units are, in turn, connected with each other via bridging sulfate anions, leading to the formation of infinite [Cu{H3C(C5H4N)}4SO4]n zigzag chains along [001]. The completed coordination spheres of the Cu2+ ions are slightly distorted octa\u00adhedral. The axial Cu\u2014O bonds are elongated [average length = 2.42\u2005(4)\u2005\u00c5] compared to the equatorial Cu\u2014N bonds [average length = 2.043\u2005(2)\u2005\u00c5]. The inter\u00adstitial space between the chains is filled with uncoordinated water mol\u00adecules that consolidate the structure through O\u2014H\u22efO hydrogen bonding. One of the five crystallographically independent solvent water mol\u00adecules is partially occupied with an occupancy factor of 0.396\u2005(4). Due to hydrogen bonding between symmetry-equivalent water mol\u00adecules across inversion centers, several of the water H atoms are disordered in 1:1 ratios over mutually exclusive positions. The crystal under investigation was found to be non-merohedrally twinned in a 0.789\u2005(1):0.211\u2005(1) ratio by a 180\u00b0 rotation around the reciprocal b axis.The structure of the title compound, {[Cu(SO DOI: 10.1107/S1600536811006325/wm2458Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Cd\u2014S and Cd\u2014O bond lengths around the Cd atoms in the bis\u00ad(thio\u00adurea) cations are in the ranges 2.580\u2005(4)\u20132.599\u2005(4) and 2.323\u2005(8)\u20132.421\u2005(9)\u2005\u00c5, respectively, and the S atoms are in a cis orientation. In the tris\u00ad(thio\u00adurea) cations, the corresponding bond lengths around the Cd atoms are slightly longer and are in the ranges 2.559\u2005(4)\u20132.706\u2005(3) and 2.303\u2005(7)\u20132.480\u2005(10)\u2005\u00c5, respectively, and the S atoms are in a fac disposition. The crystal structure features numerous N\u2014H\u22efO, N\u2014H\u22efN, O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds. Two O atoms of a sulfate anion were found to be disordered over two orientations in a 0.620\u2005(9):0.380\u2005(9) ratio. The crystal studied was a racemic twin with BASF = 0.17\u2005(5)The title compound, [Cd(CH DOI: 10.1107/S1600536812026682/hb6807Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The [ZnCl4]2\u2212 anions have a distorted tetra\u00adhedral geometry. Weak inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions exist between neighbouring aromatic rings of the cations with a centroid\u2013centroid distance of 3.712\u2005(7)\u2005\u00c5.The asymmetric unit of the title compound, (C DOI: 10.1107/S1600536811005691/cv5049Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Ni\u2014O bond lengths range from 2.004\u2005(2) to 2.106\u2005(2)\u2005\u00c5, while the Ni\u2014N bond lengths are 2.038\u2005(2) and 2.0465\u2005(19)\u2005\u00c5. The cis bond angles range from 78.64\u2005(8) to 97.30\u2005(8)\u00b0, with the former being due to the small bite of the amino\u00adalcohol ligand, while the trans bond angles range from 167.86\u2005(8) to 171.23\u2005(8)\u00b0. One of the alcohol H atoms forms a hydrogen bond with the dimethyl\u00adformamide (DMF) solvent mol\u00adecule, while the other links mol\u00adecules into chains along the b axis through inter\u00admolecular O\u2014H\u22efO hydrogen bonds. There are bifurcated C\u2014H\u22efO inter\u00adactions involving one of the nitro groups between parallel stacks of mol\u00adecules in the b-axis direction.In the title compound, [Ni(C DOI: 10.1107/S1600536811031229/jj2096Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "O\u2014H\u22efO hydrogen bonds connect adjacent complex mol\u00adecules into dimers. C\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings [centroid\u2013centroid distances = 3.700\u2005(2) and 3.845\u2005(2)\u2005\u00c5] are also present.In the title complex, [Co(C DOI: 10.1107/S1600536811029461/hy2451Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The RuII atom has a distorted octa\u00adhedral geometry due to the restricted bite angle [157.7\u2005(3)\u00b0] of the two mer-arranged N,N\u2032,N\u2032\u2032-tridendate ligands, viz. 2,2\u2032:6\u2032,2\u2032\u2032-terpyridine (tpy) and 4\u2032-ethynyl-2,2\u2032:6\u2032,2\u2032\u2032-terpyridine (tpy\u2032), which are essentially perpendicular to each other, with a dihedral angle of 87.75\u2005(12)\u00b0 between their terpyridyl planes. The rod-like acetyl\u00adene group lies in the same plane as its adjacent terpyridyl moiety, with a maximum deviation of only 0.071\u2005(11)\u2005\u00c5 from coplanarity with the pyridine rings. The mean Ru\u2014N bond length involving the outer N atoms trans to each other is 2.069\u2005(6)\u2005\u00c5 at tpy and 2.070\u2005(6)\u2005\u00c5 at tpy\u2032. The Ru\u2014N bond length involving the central N atom is 1.964\u2005(6)\u2005\u00c5 at tpy and 1.967\u2005(6)\u2005\u00c5 at tpy\u2032. Two of the three counter anions were refined as half-occupied.The title heteroleptic II\u2013terpyridine complex containing the {Ru(tpy\u2013C\u00a0C)} fragment, see: Ruben et al. (C17H11N3)](PF6)2\u00b72C2H3N = 0.084wR(F2) = 0.228S = 1.257496 reflections596 parametersH-atom parameters constrainedmax = 1.75 e \u00c5\u22123\u0394\u03c1min = \u22120.94 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536812051227/sj5287Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The compound exhibits a three-dimensional supra\u00admolecular structure composed of the complex mol\u00adecules and lattice water mol\u00adecules, which are linked together by inter\u00admolecular O\u2014H\u22efO hydrogen bonds and partly overlapping \u03c0\u2013\u03c0 inter\u00adactions between the pyridine and benzene rings [centroid\u2013centroid distances = 3.922\u2005(2) and 3.921\u2005(2)\u2005\u00c5]. One of the lattice water mol\u00adecules is disordered over two positions in an occupancy ratio of 0.521\u2005(6):0.479\u2005(6).In the title compound, [Ni(C DOI: 10.1107/S1600536811032090/hy2454Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angle between the pyridine rings is 6.3\u2005(2)\u00b0. The N\u2014C\u2014C\u2014N torsion angle is 177.5\u2005(2)\u00b0. In the dianion, the CrVI ions are in a slightly distorted tetra\u00adhedral coordination environment and the bond angles at the independent CrVI ions are in the ranges 105.93\u2005(10)\u2013110.60\u2005(11) and 107.35\u2005(11)\u2013111.07\u2005(12)\u00b0. The Cr\u2014O\u2014Cr angle is 127.96\u2005(12)\u00b0. The crystal used was an inversion twin with refined components of 0.510\u2005(19) and 0.490\u2005(19).In the cation of the title salt, (C DOI: 10.1107/S1600536812005430/lh5410Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CuII atom is five-coordinated in a distorted square-pyramidal environment by two phenolate O atoms and two imine N atoms of the tetra\u00addentate Schiff base anion in the basal plane and one water mol\u00adecule in the apical position. Because of symmetry, the pyridine N atom and the corresponding C atom at the 4-position of the pyridine ring are disordered. The crystal packing can be described as being composed of alternating layers stacked along [001]. Intra\u00admolecular C\u2014H\u22efN and inter\u00admolecular C\u2014H\u22efO and O\u2014H\u22efO hydrogen-bonding inter\u00adactions, as well as C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions [shortest centroid\u2013centroid distance = 3.799\u2005(8)\u2005\u00c5 and inter\u00adplanar distance = 3.469\u2005(2)\u2005\u00c5] are observed.Mol\u00adecules of the title compound, [Cu(C DOI: 10.1107/S1600536812031273/wm2656Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The two pyridine rings coordinating to the AgI atom are almost perpendicular to each other [dihedral angle = 87.73\u2005(10)\u00b0]. Inter\u00admolecular Ag\u22efO inter\u00adactions [3.149\u2005(3) and 2.686\u2005(3)\u2005\u00c5], N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions between the cyclic dimers and the anions or the solvent mol\u00adecules lead to the formation of a three-dimensional supra\u00admolecular network.In the binuclear title compound, [Ag I coordination polymers with symmetrical dipyridyl ligands, see: Lee et al. 2](ClO4)2\u00b72C2H6OS = 0.033wR(F2) = 0.086S = 1.023367 reflections217 parametersH-atom parameters constrainedmax = 0.61 e \u00c5\u22123\u0394\u03c1min = \u22120.65 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, New_Global_Publ_Block. DOI: 10.1107/S1600536813026585/sj5353Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the anion, the two pyridine rings are inclined to one another by 87.75\u2005(13)\u00b0. Two types of robust O\u2014H\u22efO hydrogen bond synthons, viz. R 2 2(16) and R 6 6(42), link the anions to form a two-dimensional network parallel to the bc plane. Furthermore, O\u2014H\u22efO, N\u2014H\u22efO, N\u2014H\u22efN and weak C\u2014H\u22efO hydrogen bonds connect the two dimensional networks, forming a three-dimensional structure. In the crystal, there are also C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances of 3.5554\u2005(18) and 3.7681\u2005(18)\u2005\u00c5], and C=O\u22ef\u03c0 inter\u00adactions [O\u22efcentroid distance = 3.117\u2005(2)\u2005\u00c5] present. One of the three crystal water molecules shows an occupancy of 0.35.In the title compound, (C DOI: 10.1107/S1600536811052445/lh5386Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CoII ion is six coordinate, with distorted octa\u00adhedral geometry, ligated by two N atoms and two O atoms from a 2,2\u2032-bis\u00ad(5-carboxy-1H-imidazole-4-carboxyl\u00adate) dianion, one N atom from a pyridine mol\u00adecule and one coordinating water mol\u00adecule. The Co\u2014O bond lengths range from 2.076\u2005(2) to 2.1441\u2005(15)\u2005\u00c5, while the Co\u2014N bond lengths are 2.138\u2005(3) and 2.1515\u2005(17)\u2005\u00c5. A two-dimensional network of N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds stabilizes the crystal packing. There are \u03c0\u2013\u03c0 inter\u00adactions between the bipyridine and imidazole rings [centroid\u2013centroid distance = 3.7694\u2005(4)\u2005\u00c5]. The propane-1,3-diyl group is disordered over two conformations, with refined occupancies of 0.755\u2005(8) and 0.245\u2005(8).In the title compound, [Co(C DOI: 10.1107/S1600536812029856/pk2422Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CoII ion is situated on an inversion centre and is coordinated by two O and two N atoms of two symmetry-related 1,4-pyrazine-2,3-dicarboxyl\u00adate ligands and two water mol\u00adecules and has a disorted octa\u00adhedral coordination environment. The asymmetric unit also contains four water mol\u00adecules. In the crystal, extensive inter\u00admolecular classical N\u2014H\u22efO, O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.490\u2005(1)\u2005\u00c5] connect the various components, forming a three-dimensional network.The title compound, (C DOI: 10.1107/S1600536811001127/bt5444Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Fe atom is slightly closer to the substituted cyclo\u00adpenta\u00addienyl ring, with an Fe\u22efcentroid distance of 1.639\u2005(2)\u2005\u00c5, compared with 1.645\u2005(2)\u2005\u00c5 for the unsubstituted ring. The C=N double bond is essentially coplanar with the substituted cyclo\u00adpenta\u00addienyl ring with a deviation of 10.3\u2005(1)\u00b0. The angle formed by the C=N double bond and the naphthal\u00adene ring system is 47.1\u2005(1)\u00b0. The C\u2014N=C\u2014C torsion angle is 177.32\u2005(5)\u00b0.In the title mol\u00adecule, [Fe(C DOI: 10.1107/S1600536808026330/lh2673Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atom in the title compound, [ZnCl2(C14H13N3)], is coordinated by a Cl2N2 donor set defined by quinoline and pyrazole N atoms of the chelating ligand and two Cl atoms. Distortions from the ideal tetra\u00adhedral geometry relate to the restricted bite angle of the chelating ligand [N\u2014Zn\u2014N = 78.54\u2005(12)\u00b0]. In the crystal, mol\u00adecules are connected into a three-dimensional structure by C\u2014H\u22efCl inter\u00adactions, involving both Cl atoms, and \u03c0\u2013\u03c0 inter\u00adactions that occur between the pyrazole ring and each of the pyridine and benzene rings of the quinoline residue [inter\u00adcentroid distances = 3.655\u2005(2) and 3.676\u2005(2)\u2005\u00c5].The Zn DOI: 10.1107/S1600536812014390/hg5206Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angles between the imidazole and triazole rings in the cations are 44.7\u2005(3) and 89.4\u2005(3)\u00b0. The BiIII ion is coordinated by six chloride ligands in a slightly distorted octa\u00adhedral geometry. In each cation, an intra\u00admolecular N\u2014H\u22efS hydrogen bond is observed. In the crystal, N\u2014H\u22efCl, N\u2014H\u22efS, N\u2014H\u22efO, O\u2014H\u22efCl, O\u2014H\u22efS and O\u2014H\u22efO hydrogen bonds connect the components into a three-dimensional network. In addtion, \u03c0\u2013\u03c0 stacking inter\u00adactions between inversion-related triazole rings are observed, with a centroid\u2013centroid distance of 3.322\u2005(3)\u2005\u00c5The asymmetric unit of the title hydrated salt, (C DOI: 10.1107/S1600536813016449/lh5614Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angle between the mean planes of the coordination rings is 9.15\u2005(12)\u00b0 while the dihedral angle between the mean planes of the two aromatic rings is 3.48\u2005(16)\u00b0. In the crystal, pairs of inter\u00admolecular C\u2014H\u22efO hydrogen bonds link neighboring mol\u00adecules into a chain along the a axis. The crystal structure is further stabilized by \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.883\u2005(2)\u2005\u00c5].In the title compound, [Ni(C DOI: 10.1107/S1600536811022732/kj2181Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In each mol\u00adecule, the metal atom is coordinated by an N,N\u2032,N\u2032\u2032-tridenate Schiff base and two Cl atoms in a distorted square-pyramidal geometry. The two mol\u00adecules differ little in their geometry, but more in their inter\u00admolecular inter\u00adactions. In the crystal, adjacent mol\u00adecules are connected via C\u2014H\u22efCl inter\u00adactions into a three-dimensional supra\u00admolecular structure. The network is supplemented by \u03c0\u2013\u03c0 inter\u00adactions formed between the aromatic rings of pairs of the symmetry-related mol\u00adecules [centroid\u2013centroid distances = 3.6255\u2005(10) and 3.7073\u2005(10)\u2005\u00c5]. The crystal lattice contains void spaces with a size of 52\u2005\u00c53.The asymmetric unit of the title compound, [ZnCl For the al. 1999; Sun ] = 0.021 wR(F 2) = 0.052 S = 1.04 6294 reflections313 parametersH-atom parameters constrainedmax = 0.36 e \u00c5\u22123 \u0394\u03c1min = \u22120.28 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811025669/om2444Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Cu\u2014O distances involving the four O atoms in the square plane are in the range 1.9501\u2005(11)\u20131.9677\u2005(11)\u2005\u00c5, with the Cu atom lying nearly in the plane [deviation = 0.0726\u2005(2)\u2005\u00c5]. The axial O atom occupies the peak position with a Cu\u2014O distance of 2.885\u2005(12)\u2005\u00c5, which is significantly longer than the rest of the Cu\u2014O distances. Each 1,8-nap ligand acts as bridge, linking two CuII atoms into a dinuclear structure. Inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen-bonding inter\u00adactions consolidate the structure.In the centrosymmetric dinuclear title complex, [Cu DOI: 10.1107/S1600536810028497/pv2300Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Both macrocycles exhibit a planar conformation with average deviation from the least-squares-plane of the 24 macrocycle atoms of \u039424 = 0.043\u2005\u00c5 for the first mol\u00adecule and 0.026\u2005\u00c5 for the mol\u00adecule located on an inversion center. The average Ni\u2014N bond lengths are 1.955\u2005(2) and 1.956\u2005(2)\u2005\u00c5 in the two mol\u00adecules. The mol\u00adecules form \u03c0\u2013\u03c0 dimers of inter\u00admediary strength with a mean plane separation of 3.36\u2005(2)\u2005\u00c5.The title compound, [Ni(C DOI: 10.1107/S1600536812035726/rn2107Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The MnII ion and two coordinated water mol\u00adecules lie on a twofold rotation axis. The water mol\u00adecules are involved in O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds with the triazole N atoms and carboxyl\u00adate O atoms, yielding a three-dimensional supra\u00admolecular network. \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings [centroid\u2013centroid distance = 3.836\u2005(9)\u2005\u00c5] are observed.In the title compound, [Mn(C DOI: 10.1107/S1600536811025335/hy2445Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, inter\u00admolecular N\u2014H\u22efCl and C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 contacts between the pyridine rings [centroid\u2013centroid distance = 3.5419\u2005(19)\u2005\u00c5] result in the formation of a supra\u00admolecular structure.In the anion of the title compound, (C DOI: 10.1107/S160053681202853X/hy2560Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "For example, the torsion angles N\u2014C\u2014C\u2014C involving the N-benzyl moieties are 57.3\u2005(7) and 11.6\u2005(8)\u00b0 in one mol\u00adecule and 76.5\u2005(7) and 97.4\u2005(7)\u00b0 in the other. In each mol\u00adecule, the P atom exhibits a distorted tetra\u00adhedral conformation [the bond angles at P are in the ranges 104.7\u2005(2)\u2013115.2\u2005(2) and 105.1\u2005(2)\u2013115.1\u2005(2)\u00b0 in the two molecules], and the phosphoryl group and the N\u2014H group adopt an anti orientation with respect to one another. In the crystal, mol\u00adecules are linked via N\u2014H\u22efO(P) hydrogen bonds, forming a chain parallel to the a axis.The asymmetric unit of the title compound, C DOI: 10.1107/S1600536811046046/nc2247Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal packing is stabilized by O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the aromatic rings of pyridine-2,5-dicarb\u00adoxy\u00adlate with centroid\u2013centroid distances of 3.6166\u2005(13)\u2005\u00c5.In the polymeric title compound, [Ca(C DOI: 10.1107/S1600536810054334/bt5441Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The porphyrin N atoms occupy the equatorial positions while the axial positions are occupied by the O atoms of the methane\u00adsulfonate anions. The phenyl rings make dihedral angles of 77.02\u2005(13) and 87.89\u2005(14)\u00b0 with the porphyrin ring. Of the three solvent chloro\u00adform mol\u00adecules, one is disordered over a twofold rotation axis. In the crystal a three-dimensional assembly is accomplished via C\u2014H\u22efO hydrogen bonds between the H atoms of the phenyl groups in the porphyrin ring and the O atoms of the methane\u00adsulfonate ligands.In the crystal structure of the title compound, [Sn(C For the al. 2004, 2009 \u25b6.44H28N4)(CH3O3S)2]\u00b73CHCl3 = 0.033wR(F2) = 0.088S = 1.045192 reflections331 parametersH-atom parameters constrainedmax = 0.91 e \u00c5\u22123\u0394\u03c1min = \u22120.86 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812015875/wm2606Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angle between the phenyl ring and the metal-bound pyridine ring is 50.3\u2005(4)\u00b0 for each 2-phenyl\u00adpyridine ligand. This arranges the phenyl ring from one ligand in the complex above the pyridine ring of the other resulting in an intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adaction, with a centroid\u2013centroid distance of 3.6796\u2005(17)\u2005\u00c5. Weak C\u2014H\u22efCl hydrogen bonds stabilize the crystal packing, linking mol\u00adecules into chains along the c axis.In the title compound, [ZnCl DOI: 10.1107/S1600536812010616/sj5207Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "III atom of the title compound, [Ir(C11H8N)2Cl(CH3CN)], displays a distorted octa\u00adhedral coordination. The pyridyl groups are in trans positions [N\u2014Ir\u2014N = 173.07\u2005(10)\u00b0], while the phenyl groups are trans with respect to the acetonitrile and chloride groups [C\u2014Ir\u2014N = 178.13\u2005(11) and C\u2014Ir\u2014Cl = 176.22\u2005(9)\u00b0]. The pyridyl\u00adphenyl groups only show a small deviation from planarity, with the dihedral angle between the planes of the two six-membered rings in each pyridyl\u00adphenyl group being 5.6\u2005(2) and 5.8\u2005(1)\u00b0. The crystal packing shows inter\u00admolecular C\u2014H\u22efCl, C\u2014H\u22ef\u03c0(acetonitrile) and C\u2014H\u22ef\u03c0(pyridyl\u00adphen\u00adyl) contacts.The Ir DOI: 10.1107/S1600536811049373/tk5023Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Mn\u2014O distances range from 2.151\u2005(2) to 2.5093\u2005(19)\u2005\u00c5, while the Mn\u2014N distances are 2.226\u2005(2) and 2.248\u2005(2)\u2005\u00c5. Each 5-ferrocenyl\u00adbenzene-1,5-dicarboxyl\u00adate anion links to two MnII ions, resulting in a chain along the b axis. A three-dimensional network of N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds helps to stabilize the crystal packing.In the title coordination polymer, {[FeMn(C DOI: 10.1107/S1600536811022781/fj2428Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, the mol\u00adecules are linked via weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds [3.334\u2005(5)\u2005\u00c5] and there are also short inversion-related intermolecular Br\u22efBr contacts [3.4263\u2005(6)\u2005\u00c5]In the title complex, [Co(C DOI: 10.1107/S1600536810032162/zs2053Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The NiII ion is situated on an inversion centre. The Ni\u2014N distances range from 2.0589\u2005(19) to 2.0757\u2005(19)\u2005\u00c5, intra-ligand N\u2014Ni\u2014N angles range from 84.50\u2005(8) to 85.15\u2005(8)\u00b0, and adjacent inter-ligand N\u2014Ni\u2014N angles range between 94.85\u2005(8) and 95.50\u2005(8)\u00b0. In the crystal, O\u2014H\u22efO hydrogen bonds between methanol solvent mol\u00adecules and tri\u00adfluoro\u00admethane\u00adsulfonate anions are observed.In the title salt, [Ni(C DOI: 10.1107/S1600536813024653/lh5647Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The EuIII ion is coordinated by nine O atoms from two H2bdc ligands, two sulfate anions and three water mol\u00adecules, displaying a bicapped trigonal prismatic geometry. The carboxyl\u00adate groups of the H2bdc ligands and the sulfate anions link the EuIII ions, forming a chain along [010]. These chains are further connected by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the imidazole and benzene rings into a three-dimensional supra\u00admolecular network.In the title coordination polymer, H-benzimidazole-5,6-dicarboxyl\u00adate complexes, see: Wang et al. 2(SO4)2(H2O)6]\u00b76H2O = 0.036 wR(F 2) = 0.097 S = 1.02 2841 reflections248 parametersH atoms treated by a mixture of independent and constrained refinementmax = 1.82 e \u00c5\u22123 \u0394\u03c1min = \u22122.51 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 global. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The angles around the CuII atom vary between 85.32\u2005(6) and 94.68\u2005(6)\u00b0. The Cu\u2014O bond distances between the CuII atom and the O atoms vary between 1.9424\u2005(14) and 2.3229\u2005(15)\u2005\u00c5. The layers inter\u00addigitate via face-to-face aromatic inter\u00adactions [3.6490\u2005(8)\u2005\u00c5] between coumarin moieties such that the inter\u00adlayer separation is 10.460\u2005(2)\u2005\u00c5, i.e. the length of the c axis. O\u2014H\u22efO hydrogen bonds between the H atoms of coordinated water mol\u00adecules and the O atoms of carboxyl\u00adate groups link the complex mol\u00adecules into layers parallel to the ab plane.In the title compound, [Cu(C DOI: 10.1107/S1600536811018708/zk2008Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions and short inter\u00admolecular I\u22efO contacts [3.142\u2005(2)\u2005\u00c5] are observed.In the title compound, C DOI: 10.1107/S1600536810033581/nc2194Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The average Ru\u2014C distance to the penta\u00admethyl\u00adcyclo\u00adpenta\u00addienyl (Cp*) group is 2.19\u2005(3)\u2005\u00c5, and 2.21\u2005(1)\u2005\u00c5 to the ortho, meta and para C atoms of the arene ring. The Ru\u2014Cipso bond length of 2.272\u2005(3)\u2005\u00c5 is significantly longer, reflecting movement of the Ru atom away from the C atoms with electronegative substituents attached. The amide H atom in the cation forms an inter\u00admolecular N\u2014H\u22efO hydrogen bond with the carbonyl O atom of the acetone solvent mol\u00adecule. A C\u2014H\u22efO inter\u00adaction also occurs.The title complex, [Ru(C DOI: 10.1107/S1600536811031655/ng5208Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds result in a supra\u00admolecular layer parallel to (101). These layers are connected by \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings [centroid\u2013centroid distance = 3.891\u2005(2)\u2005\u00c5].In the title complex, [Cu(NO DOI: 10.1107/S160053681103529X/hy2463Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angle between the substituted benzene rings is 29.95\u2005(16)\u00b0. In the crystal, mol\u00adecules are linked along the b axis, forming individual dimers through C\u2014H\u22efO inter\u00adactions. The crystal structure is further stabilized by inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.6131\u2005(17)\u2005\u00c5].In the title Schiff base complex, [Cu(C DOI: 10.1107/S160053681200195X/hp2025Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The geometry around the ZnII atoms is distorted trigonal\u2013bipyramidal, supported by the N2O2 donor atoms of the tetradentate Schiff base and a coordinating water mol\u00adecule. The dihedral angles between the benzene rings in the two mol\u00adecules are 34.10\u2005(15)\u2005\u00c5 and 30.61\u2005(15)\u2005\u00c5. In the crystal, neighbouring independent mol\u00adecules are linked by pairs of O\u2014H\u22efO hydrogen bonds, forming dimers with R22(6) ring motifs, and by O\u2014H\u22efCl hydrogen bonds. There are short Cl\u22efCl contacts present, and mol\u00adecules are also linked by C\u2014H\u22efO and \u03c0\u2013\u03c0 [centroid\u2013centroid distance = 3.671\u2005(2)\u2005\u00c5] inter\u00adactions.The asymmetric unit of the title compound, [Zn(C DOI: 10.1107/S1600536812025986/su2451Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Two PbII atoms are bridged by the Hbtc ligands, forming a discrete centrosymmetric dinuclear complex. Inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the pyridine and imidazole rings, and between the pyridyl rings of the L ligands [centroid\u2013centroid distances = 3.600\u2005(6) and 3.732\u2005(6)\u2005\u00c5] lead to a three-dimensional supra\u00admolecular structure.In the title compound, [Pb DOI: 10.1107/S1600536810045812/hy2360Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atom in the title salt, [Cu(C10H24N4)(H2O)2][CH3(CH2)8CO2]2\u00b72H2O, is chelated by the four N atoms of the 1,4,8,11-tetra\u00adaza\u00adcyclo\u00adtetra\u00addecane (cyclam) ligand and is coordinated by two water mol\u00adecules in a Jahn\u2013Teller-type tetra\u00adgonally distorted octa\u00adhedral geometry. The CuII atom lies on a center of inversion. The cations, anions and uncoordinated water mol\u00adecules are linked by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, forming a layer structure parallel to (001).The Cu DOI: 10.1107/S1600536810025699/bt5286Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "I atom in the title compund, [Cu(C7H2Br3O2)(C19H17P)2], is located on a twofold rotation axis; the 3,5,7-tribromo\u00adtropolonate anion coordinates as a bidentate ligand with a bite angle of 76.42\u2005(9)\u00b0. An intra\u00admolecular C\u2014H\u22efO inter\u00adaction occurs. Within the crystal, extensive weak C\u2014H\u22ef\u03c0 inter\u00adactions contribute to the herringbone pattern observed in the packing of the mol\u00adecules.The Cu DOI: Click here for additional data file.10.1107/S1600536812042286/ng5299Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The coordination sphere about the Na atom in each complex unit comprises five bonds to O atoms of the crown ether [Na\u2014O = 2.390\u2005(7)\u20132.466\u2005(6)\u2005\u00c5] and one to a thio\u00adsulfate O atom [Na\u2014O = 2.305\u2005(4) and 2.447\u2005(3)\u2005\u00c5].In the title complex, [Na(C DOI: 10.1107/S1600536811022252/zs2116Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials: interactive version of Fig. 1 Enhanced figure:"} +{"text": "In the crystal, O\u2014H\u22efCl and N\u2014H\u22efCl hydrogen bonds link the complex mol\u00adecules into a layer parallel to (100). \u03c0\u2013\u03c0 inter\u00adactions between the thia\u00adzole rings are observed [centroid\u2013centroid distance = 3.749\u2005(3)\u2005\u00c5].In the title complex, [CuCl DOI: 10.1107/S1600536812013037/hy2524Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angles between the central benzene ring and the two outer rings are 4.79\u2005(15) and 7.54\u2005(15)\u00b0. In the crystal, mol\u00adecules are connected through inter\u00admolecular C\u2014H\u22efO hydrogen bond, resulting in chains extending along the c axis. The crystal structure is further stabilized by inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions, with centroid\u2013centroid distances in the range 3.3760\u2005(15)\u20133.7196\u2005(17)\u2005\u00c5.In the title Schiff base complex, [Ni(C DOI: 10.1107/S1600536810051834/pv2367Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The chelate ring including the CuII atom is essentially planar, with a maximum deviation of 0.0181\u2005(17)\u2005\u00c5 for one of the coordinated N atoms. This plane forms a dihedral angle of 30.75\u2005(6)\u00b0 with the CuCl2 plane. In the crystal, each pair of adjacent mol\u00adecules is linked into a centrosymmetric dimer by N\u2014H\u22efCl hydrogen bonds. The crystal structure is stabilized by inter\u00admolecular C\u2014H\u22efN and C\u2014H\u22efCl hydrogen bonds and weak slipped \u03c0\u2013\u03c0 stacking inter\u00adactions between symmetry-related mol\u00adecules, with an inter\u00adplanar separation of 3.439\u2005(19)\u2005\u00c5 and a centroid\u2013centroid distance of 3.581\u2005(19)\u2005\u00c5.In the title complex, [CuCl DOI: 10.1107/S1600536811004375/fj2386Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by \u03c0\u2013\u03c0 stacking inter\u00adactions [shortest centroid\u2013centroid distance = 3.5537\u2005(18)\u2005\u00c5].In the title complex, [Cd(NO DOI: 10.1107/S160053681002550X/tk2681Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Besides two pyridine-2,5-dicarboxyl\u00adate anions, one bidentate 2,2\u2032-bipyridine and one water mol\u00adecule coordinate to the Co cation, completing a distorted octa\u00adhedral coordination geometry. Within the dinuclear mol\u00adecule, \u03c0\u2013\u03c0 stacking occurs between parallel pyridine rings with centroid\u2013centroid distances of 3.802\u2005(2)\u2005\u00c5. The crystal structure contains extensive O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions.In the centrosymmetric title compound, [Co DOI: 10.1107/S1600536811003059/xu5145Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Zn(TPP) complex exhibits a nearly planar conformation of the porphyrin core [maximum deviation = 0.106\u2005(2)\u2005\u00c5] with an average Zn\u2014N distance of 2.047\u2005(2)\u2005\u00c5. The title compound is considered as a one-dimensional polymer along [010], in which the Zn(TPP) moiety is linked to the closest O atoms of two symmetry-related 18-crown-6 mol\u00adecules with a Zn\u2014O distance of 2.582\u2005(1)\u2005\u00c5, completing a distorted octahedral coordination environment of the metal ion. The chains are mainly sustained by weak C\u2014H\u22ef\u03c0 inter\u00adactions. An ethyl\u00adene group of the 18-crown-6 mol\u00adecule is disordered over three sites with occupancies of 0.50, 0.25 and 0.25.In the title compound, [Zn(C DOI: 10.1107/S1600536813018126/hy2633Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The N atoms are cis to each other, while the water O atoms and ligand O atoms are mutually trans. The crystal structure is stabilized by a network of O\u2014H\u22efO, O\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.730\u2005(3) and 3.652\u2005(3)\u2005\u00c5] between the 5-methyl\u00adpyrazine-2-carboxyl\u00adate ligands. The structure is isotypic with the manganese analog.In the title compound, [Cd(C DOI: 10.1107/S1600536811035045/bt5630Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit is composed of six TPPI mol\u00adecules, eight TFTIB mol\u00adecules and two methanol mol\u00adecules, overall 16 constituents. The formation of the architecture is essentially guided by a number of C\u2014I\u22efI\u2212 halogen bonds (XB), whose lengths are in the range 3.276\u2005(1)\u20133.625\u2005(1)\u2005\u00c5. Layers of supra\u00admolecular polyanions are formed parallel to (10-1) wherein iodide anions function as penta-, tetra- or bidentate XB acceptors. The structure is not far from being P21/n, but the centrosymmetry is lost due to a different conformation of a single couple of cations and the small asymmetry in the formed supra\u00admolecular anion. One methanol mol\u00adecule is hydrogen bonded to an iodide anion, while the second is linked to the first one via an O\u2014H\u22efO contact. This second methanol mol\u00adecule is more loosely pinned in its position than the first and presents very high anisotropic displacement parameters and a seeming shortening of the C\u2014O bond length. The crystal studied was refined as a perfect inversion twin.The crystallization of a 1:1 molar solution of 1,3,5-tri\u00adfluoro-2,4,6-di\u00adiodo\u00adbenzene (TFTIB) and tetra\u00adphenyl\u00adphosponium iodide (TPPI) from methanol produced tetra\u00adgonal needles of pure TPPI and tabular pseudo-hexa\u00adgonal truncated bipyramids of the title compound, 3C \u00c5b = 22.001 (4) \u00c5c = 28.260 (5) \u00c5\u03b2 = 92.49 (2)\u00b0V = 10704 (3) \u00c53Z = 4K\u03b1 radiationMo \u22121\u03bc = 4.45 mmT = 90 K0.34 \u00d7 0.20 \u00d7 0.12 mmBruker SMART APEX diffractometerSADABS; Bruker, 1998)Tmin = 0.614, Tmax = 1.000Absorption correction: multi-scan (105822 measured reflections28547 independent reflectionsI > 2\u03c3(I)26385 reflections with Rint = 0.047R[F2 > 2\u03c3(F2)] = 0.040wR(F2) = 0.081S = 1.0928547 reflections2304 parameters13 restraintsH-atom parameters constrainedmax = 2.46 e \u00c5\u22123\u0394\u03c1min = \u22120.89 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SIR2002 ORTEP-3 for Windows global, I. DOI: Click here for additional data file.10.1107/S1600536813012397/im2426Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The mol\u00adecules are stabilized by C\u2014H\u22efCl hydrogen bonds. In the crystal, inter\u00admolecular C\u2014H\u22efCl and C\u2014H\u22efS hydrogen bonds with R 2 2(8), R 4 2(8) and R 2 2(6) ring motifs generate a polymeric network.In the centrosymmetric title compound, [Co DOI: 10.1107/S1600536811013067/si2349Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There are hydrogen-bonding inter\u00adactions between the secondary amine H atom and the phenolate O atom, as well as C\u2014H\u22efO inter\u00adactions, which result in the dihedral angle between the aromatic phenyl ring of the 2-formyl\u00adphenolate moiety and the pyridine ring being 80.23\u2005(4)\u00b0. In the packing, there are both C\u2014H\u22ef\u03c0 inter\u00adactions, which link the mol\u00adecules into chains along the b axis, and offset \u03c0\u2013\u03c0 inter\u00adactions involving both the pyridine and phenyl rings of the 2-BAP ligands [centroid\u2013centroid distances = 4.0100\u2005(8)\u2005\u00c5 for the pyridine rings and 3.6601\u2005(8) and 4.8561\u2005(8)\u2005\u00c5 for the phenyl rings].In the title complex, [Ni(C DOI: 10.1107/S1600536811001425/zl2338Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular network. \u03c0\u2013\u03c0 stacking inter\u00adactions between the imidazole rings [centroid\u2013centroid distances = 3.5188\u2005(15) and 3.6687\u2005(15)\u2005\u00c5] further stabilize the structure.In the title compound, [Mn(C DOI: Click here for additional data file.10.1107/S1600536813004091/hy2616Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The two complex cations have similar conformations with the butterfly-like Fe4S4 core surrounded by two penta\u00admethyl\u00adcyclo\u00adpenta\u00addienyl ligands and the S atoms of two dithiol\u00adate ligands. In each Fe4S4 core, there are four short Fe\u2014Fe and two long Fe\u22efFe contacts, suggesting bonding and non-bonding inter\u00adactions, respectively. The Fe\u2014S distances range from 2.1287\u2005(13) to 2.2706\u2005(16)\u2005\u00c5 for one and from 2.1233\u2005(13) to 2.2650\u2005(16)\u2005\u00c5 for the other Fe4S4 core. The Fe\u2014S distances involving the dithiol\u00adate ligands are in a more narrow range [2.1764\u2005(16)\u20132.1874\u2005(13)\u2005\u00c5 for one and 2.1743\u2005(14)\u20132.1779\u2005(16)\u2005\u00c5 for the other cation]. There are no significant inter\u00adactions between cations and anions.The asymmetric unit of the title compound, [Fe DOI: Click here for additional data file.10.1107/S1600536813007514/wm2732Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The metal atoms are bridged by 1,2-bis\u00ad[4-(pyridin-3-yl)pyrimidin-2-ylsulfan\u00adyl]ethane ligands, giving a polymeric chain extending along the b axis. Adjacent chains related by an inversion center are further bridged by Cd\u2014O bonds formed between the O atom of one of the acetate ligands and the metal atom. The five Cd\u2014O bond lengths are in the range 2.329\u2005(3)\u20132.485\u2005(3)\u2005\u00c5. There are \u03c0\u2013\u03c0 stacking inter\u00adactions between the aromatic rings of adjacent polymeric chains, the centroid\u2013centroid distances being 3.556\u2005(3) and 3.698\u2005(3)\u2005\u00c5, organizing the chains into a three-dimensional framework. This framework is additionally stabilized by extensive O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonding between water mol\u00adecules and the ligands.The title compound, {[Cd(CH DOI: 10.1107/S1600536811046794/gk2423Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ZnII ion is displaced by 0.586\u2005\u00c5 in the direction of the chloride ligand from the atoms forming the square plane. In the crystal, the components are linked by inter\u00admolecular O\u2014H\u22efO hydrogen bonds, generating chains along the b axis.In the title compound, [Zn(C Cl(C9H7NO)]\u00b7CH4O = 0.034 wR(F 2) = 0.082 S = 1.05 3806 reflections244 parameters2 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.47 e \u00c5\u22123 \u0394\u03c1min = \u22120.42 e \u00c5\u22123 \u0394\u03c1 CrysAlis PRO used to solve structure: SHELXS97 global, I. DOI: 10.1107/S160053681103337X/lh5311Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dicationic complexes are arranged in a staggered configuration. The torsion angle subtended by the 1-methyl\u00adimidazole ring relative to the terpyridine ring is 114.9\u2005(5)\u00b0. Inter\u00admolecular C\u2014H\u22efO inter\u00adactions between the perchlorate anions and the H atoms of the terpy ligand are observed. Consideration of related phenyl\u00adbipyridyl complexes of platinum(II), which are monocationic, leads to the conclusion that the electrostatic repulsion between the dicationic chelates prevents the formation of Pt\u22efPt inter\u00adactions. These inter\u00adactions are a common feature associated with the monocationic species.The reaction between [Pt(terpy)Cl]\u00b72H For the al. 1996 and for al. 2007. For stu al. 2007. For a c al. 2008. For Pt\u22ef al. 1997; Field e al. 2003; Jaganyi al. 2003. 15H11N3)(C4H6N2)](ClO4)2\u00b7CH3NO2 = 0.045 wR(F 2) = 0.118 S = 0.98 5102 reflections354 parametersH-atom parameters constrainedmax = 1.59 e \u00c5\u22123 \u0394\u03c1min = \u22122.64 e \u00c5\u22123 \u0394\u03c1 CrysAlis CCD I, global. DOI: 10.1107/S1600536811025475/fi2109Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The inter\u00adnal benzene rings, A and B, make dihedral angles of 86.72\u2005(5) and 79.22\u2005(5)\u00b0 with the naphthalene ring system. The two terminal benzene rings, C and D, of the 4-phen\u00adoxy\u00adbenzoyl groups are twisted with respect to benzene rings A and B, with dihedral angles of A/C = 62.72\u2005(8) and B/D\u00a0= 87.61\u2005(6)\u00b0. In the crystal, H atoms in the naphthalene system make two types of inter\u00admolecular C\u2014H\u22efO inter\u00adactions with the carbonyl O atom and the phenyl etheral O atom of neighbouring mol\u00adecules. Mol\u00adecules are further linked by C\u2014H\u22ef\u03c0 inter\u00adactions involving a H atom of terminal benzene ring D and the \u03c0-system of the inter\u00adnal benzene ring A, forming dimers centered about an inversion center.In the title mol\u00adecule {systematic name: (4-phenoxyphenyl)methan\u00adone}, C DOI: 10.1107/S1600536810042170/su2217Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The geometry around the NiII atom in each mol\u00adecule is distorted square planar. The dihedral angles between the two phen\u00adoxy rings in each mol\u00adecule are 17.8\u2005(4) and 36.5\u2005(4)\u00b0. The crystal packing is stabilized by weak \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.758\u2005(5)\u2005\u00c5] and C\u2014H\u22ef\u03c0 inter\u00adactions.The asymmetric unit of the title complex, [Ni(C DOI: 10.1107/S1600536811029813/su2297Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the cation, the average Sb\u2014N distance is 2.066\u2005(2)\u2005\u00c5, while the Sb\u2014Cl distances are 2.3410\u2005(11) and 2.3639\u2005(12)\u2005\u00c5. The central unit of the cation, SbN4C20, is far from being planar, with deviations of atoms from the least-squares plane ranging from \u22120.110\u2005(4) to 0.124\u2005(4)\u2005\u00c5. The Sb\u2014Cl distances in the anion, which is located about an inversion center, lie in the wide range 2.3715\u2005(13)\u20132.7489\u2005(13)\u2005\u00c5, the longest distances being between the Sb and bridging Cl atoms. The crystal structure is stabilized by inter\u00admolecular C\u2014H\u22efCl inter\u00adactions involving the cations, the anions and the solvent mol\u00adecules. The solvent mol\u00adecule is disordered over two orientations in a 0.901\u2005(13):0.099\u2005(13) ratio.The asymmetric unit of the title complex, [Sb(C Cl2][Sb2Cl8]0.5\u00b7CH2Cl2 = 0.052wR(F2) = 0.114S = 1.1112918 reflections539 parameters2 restraintsH-atom parameters constrainedmax = 1.36 e \u00c5\u22123\u0394\u03c1min = \u22120.88 e \u00c5\u22123\u0394\u03c1COLLECT global, I. DOI: 10.1107/S1600536812018351/gk2479Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "One Cl atom and one water mol\u00adecule in trans positions complete a distorted octa\u00adhedral geometry around the metal atom. In the crystal, the complex mol\u00adecules and the uncoordinated water mol\u00adecules are connected via O\u2014H\u22efN, O\u2014H\u22efO and O\u2014H\u22efCl hydrogen bonds into layers parallel to the ac plane and these are consolidated by C\u2014H\u22ef\u03c0 inter\u00adactions. The layers are further linked into a three-dimensional network through C\u2014H\u22efO inter\u00adactions.In the title compound, [Mn(C II complex of the same Schiff base, see: Ikmal Hisham et al. 2Cl(H2O)]\u00b7H2O = 0.039 wR(F 2) = 0.093 S = 0.99 6393 reflections373 parameters4 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.43 e \u00c5\u22123 \u0394\u03c1min = \u22120.32 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811026493/is2745Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit consists of one half of the mol\u00adecular unit, the remainder generated by a twofold rotation axis located along the Cl\u2014Sn\u2014Cl bonds. The SnIV atom is coordinated by two phenyl groups, two Cl atoms and two thio\u00adurea ligands in an all trans octa\u00adhedral C2Cl2S2 environment. Individual mol\u00adecules are connected through N\u2014H\u22efCl hydrogen bonds, leading to a three-dimensional network structure. Intra\u00admolecular N\u2014H\u22efCl hydrogen bonds are also present.The title compound, [Sn(C For org al. 2001; Pelleri al. 2001. For chl al. 2002; M\u00fcller al. 2008. For tin al. 1984; Sow et al. 2012; Wirth e al. 1998.6H5)2Cl2(CH4N2S)2] = 0.019wR(F2) = 0.051S = 1.062183 reflections123 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.40 e \u00c5\u22123\u0394\u03c1min = \u22120.72 e \u00c5\u22123\u0394\u03c1COLLECT used to solve structure: SIR97 I, New_Global_Publ_Block. DOI: 10.1107/S1600536813024343/wm2764Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Pt atom is located on an inversion centre and thus the asymmetric unit contains one half of the complex; the PtN2Cl2 unit is exactly planar. The dihedral angle between the PtN2Cl2 unit and the quinoline ligand is 85.1\u2005(1)\u00b0. In the crystal, the complex mol\u00adecules are stacked into columns along the b axis. In the columns, several inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the six-membered rings are present, the shortest ring centroid\u2013centroid distance being 3.733\u2005(5)\u2005\u00c5 between pyridine rings.In the title complex, Ha 2012a. For the al. 2001; Ha (201l. 2001b.2(C9H7N)2] = 0.034wR(F2) = 0.080S = 0.971569 reflections106 parametersH-atom parameters constrainedmax = 1.74 e \u00c5\u22123\u0394\u03c1min = \u22120.97 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812013608/ng5260Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CuI ion is coordinated in a distorted tetra\u00adhedral geometry by two bridging Br atoms in addition to an N and an S atom from the 2-[disulfan\u00adyl]-4,6-dimethyl\u00adpyrimidine ligand. In the crystal, \u03c0\u2013\u03c0 stacking inter\u00adactions are observed with a centroid\u2013centroid distance of 3.590\u2005(2)\u2005\u00c5.The title dinuclear complex, [Cu DOI: 10.1107/S1600536812016315/lh5449Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The PdII atom is in a distorted quadratic coordination by two P and two Cl atoms with bond lengths of 2.2414\u2005(3) and 2.2438\u2005(3)\u2005\u00c5 for Pd\u2014P, and 2.3452\u2005(3) and 2.3565\u2005(3)\u2005\u00c5 for Pd\u2014Cl. The conformation of the Pd complex is controlled by an intra\u00admolecular slipped \u03c0\u2013\u03c0 stacking inter\u00adaction between a phenyl and a cyclo\u00adpenta\u00addienyl ring with corresponding C\u22efC distances starting at 3.300\u2005(2)\u2005\u00c5 and the distance between ring centroids being 3.674\u2005(2)\u2005\u00c5. The crystal structure is stabilized by C\u2014H\u22efCl and C\u2014H\u22efO hydrogen bonds. The (CH3)2SO solvent mol\u00adecules are arranged in layers parallel to (101) and are linked in pairs by C\u2014H\u22efO inter\u00adactions. One (CH3)2SO mol\u00adecule is orientationally disordered [occupancy ratio 0.8766\u2005(17):0.1234\u2005(17)] with sulfur in two positions at both sides of its C2O triangle.The racemic title compound, [FePdCl DOI: 10.1107/S160053681103618X/gk2404Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The correct text is \"C\u03b1-model\" \"C"} +{"text": "Two N and two O atoms of the tetra\u00addentate ligand and an O atom of an ethanol ligand form the equatorial plane. The dihedral angle between the mean planes of the two benzene rings is 34.8\u2005(3)\u00b0. In the crystal, relatively strong O\u2014H\u22efO hydrogen bonds connect the complex and ethanol solvent mol\u00adecules into alternating centrosymmetric R 2 2(8) and R 4 4(16) ring motifs, forming chains along [100]. Weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds are also present.In the title compound, [U(C S-alkyl-thio\u00adsemicarbazone compounds, see: Gerbeleu & Revenko O2(C2H6O)]\u00b7C2H6O = 0.041 wR(F 2) = 0.120 S = 1.01 5467 reflections309 parameters1 restraintH-atom parameters constrainedmax = 2.48 e \u00c5\u22123 \u0394\u03c1min = \u22121.18 e \u00c5\u22123 \u0394\u03c1 CrysAlis PRO used to solve structure: SIR92 global. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit contains two Na+ cations located on a centre of symmetry and on a mirror plane, respectively, one half of a bis-phospho\u00adnate anion (the entire anion is completed by mirror symmetry), one chloride anion on a mirror plane and one water mol\u00adecule in general positions. The two Na+ cations exhibit distorted octa\u00adhedral NaCl2O4 coordination polyhedra, each consisting of two deprotonated O atoms of the bis-phospho\u00adnate anion, of two water mol\u00adecules and of two chloride anions. Strong O\u2014H\u22efO hydrogen bonds between the \u2013OH group and one of the free O atoms of the bis-phospho\u00adnate anion connect adjacent layers along [100], supported by N\u2014H\u22efCl inter\u00adactions. Intra\u00adlayer O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds are also observed.The title compound, [Na Cl(H2O)2][NaMr = 321.50Monoclinic, a = 5.53806 (4) \u00c5b = 10.50365 (8) \u00c5c = 10.2096 (1) \u00c5\u03b2 = 104.0764 (7)\u00b0V = 576.06 (1) \u00c53Z = 2K\u03b11 radiationCu \u03bb = 1.5406 \u00c5\u22121\u03bc = 6.62 mmT = 298 KFlat sheet, 8 \u00d7 8 mmSTOE Transmission STADI P diffractometerSpecimen mounting: powder loaded between two Mylar foilsData collection mode: transmissionScan method: stepGSAS = 0.02572 = 1.769\u03c74250 data points109 parameters10 restraintsH atoms treated by a mixture of independent and constrained refinementWinXPOW used to solve structure: EXPO2009 global, I. DOI: 10.1107/S1600536812018077/wm2620Isup2.rtvRietveld powder data: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The resulting SmN2O6 coordination polyhedron is a distorted square anti\u00adprism. In the crystal, C\u2014H\u22efO inter\u00adactions connect mol\u00adecules into chains along the b-axis direction. In addition, \u03c0\u2013\u03c0 stacking inter\u00adactions are observed with centroid\u2013centroid distances in the range 3.6422\u2005(13)\u20133.7329\u2005(13)\u2005\u00c5.In the title compound, [Sm(C DOI: 10.1107/S1600536813016139/ff2109Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The asymmetric unit is completed by one 4-methyl\u00adbenzoate anion, one isonicotinamide (INA) ligand and one uncoordinated water mol\u00adecule; all the ligands are monodentate. The two O and the two N atoms around the CuII ion form a slightly distorted square-planar arrangement. The dihedral angle between the carboxyl\u00adate group and the attached benzene ring is 13.86\u2005(9)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 86.08\u2005(5)\u00b0. The uncoordinated water mol\u00adecules are linked to the INA ligands by O\u2014H\u22efO hydrogen bonds. In the crystal structure, inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network.In the centrosymmetric title compound, [Cu(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2]\u00b72H2O = 0.035 wR(F 2) = 0.080 S = 1.08 3199 reflections204 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.63 e \u00c5\u22123 \u0394\u03c1min = \u22120.51 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997WinGX (Farrugia, 1999Data collection: 10.1107/S1600536810028060/su2197sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810028060/su2197Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the three components are assembled into a tape structure along the a axis by O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. Between the tapes, a \u03c0\u2013\u03c0 inter\u00adaction with a centroid\u2013centroid distance of 3.569\u2005(3)\u2005\u00c5 and a weak C\u2014H\u22efF hydrogen bond are observed.In the title compound, [Mn(C DOI: 10.1107/S1600536811049968/is2782Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II polymeric complex, [Cd(C7H4ClO2)2(C10H14N2O)(H2O)]n, the CdII cation is chelated by two chloro\u00adbenzoate anions and coordinated by two N,N-di\u00ad\u00adethyl\u00adnicotinamide (DENA) ligands and one water mol\u00adecule in a distorted NO6 penta\u00adgonal\u2013bipyramidal geometry. The CdII cations are bridged by the pyridine N atom and carbonyl O atom of the DENA ligand to form a polymeric chain running along the b axis. Inter\u00admolecular O\u2014H\u22efO hydrogen bonds between coordinating water mol\u00adecules and carboxyl\u00adate groups link adjacent chains into layers parallel to the bc plane. \u03c0\u2013\u03c0 contacts between benzene rings [shortest centroid\u2013centroid distance = 3.912\u2005(2)\u2005\u00c5] further stabilizes the crystal structure. In the mol\u00adecule, weak C\u2014H\u22efO hydrogen bonds occur between the pyridine ring and carboxyl\u00adate groups; the dihedral angles between the carboxyl\u00adate groups and adjacent benzene rings are 4.6\u2005(3) and 12.8\u2005(3)\u00b0, while the benzene rings are oriented at a dihedral angle of 1.89\u2005(13)\u00b0.In the crystal of the title Cd N,N-di\u00adethyl\u00adnicotinamide, see: Bigoli et al. 2(C10H14N2O)(H2O)] = 0.046wR(F2) = 0.093S = 1.356531 reflections326 parameters4 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.77 e \u00c5\u22123\u0394\u03c1min = \u22121.02 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S160053681301965X/xu5721Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The acesulfamate ligand is N-bound to Pd [Pd\u2014N = 2.127\u2005(2)\u2005\u00c5] with a dihedral angle of 76.35\u2005(6)\u00b0 relative to the square plane. Relatively long phen\u00adyl\u2013acesulfamate C\u2014H\u22efO and phen\u00adyl\u2013fluorine C\u2014H\u22efF inter\u00adactions consolidate the crystal packing.The title acesulfamate complex, [Pd(C DOI: 10.1107/S1600536811002911/gw2097Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the cation, the CoII atom is six-coordinated in a distorted octa\u00adhedral geometry. It bonds to two O atoms of two water mol\u00adecules, and two pairs of N atoms from two 4-amino-3,5-bis\u00ad(2-pyrid\u00adyl)-4H-1,2,4-triazole mol\u00adecules, which behave as bidentate chelating ligands. There are intra\u00admolecular N\u2014H\u22efN hydrogen bonds in the cation. In the crystal, there are a number of inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, as well as inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.657\u2005(2) and 3.847\u2005(2)\u2005\u00c5], that link the mol\u00adecules into two-dimensional networks lying parallel to the ab plane. The presence of C\u2014H\u22efO inter\u00adactions leads to the formation of a three-dimensional network.The title complex, [Co(C DOI: 10.1107/S1600536811035446/su2310Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The FeIII atom is displaced by 0.40\u2005(1)\u2005\u00c5 towards the trifluoro\u00admethane\u00adsulfonate anion from the 24-atom mean plane of the porphyrin. The average Fe\u2014Np distance is 2.044\u2005(2)\u2005\u00c5 and the Fe\u2014O distance is 2.001\u2005(2)\u2005\u00c5.The title compound, [Fe(CF DOI: 10.1107/S1600536811001395/bv2171Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The bond lengths lie within the range 2.3000\u2005(2)\u20132.326\u2005(2)\u2005\u00c5 for the Tb\u2014O bonds and 2.543\u2005(3)\u20132.553\u2005(3)\u2005\u00c5 for the Tb\u2014N bonds. The NaI atom is five-coordinated by two water O atoms and three carboxyl\u00adate O atoms in a distorted square-pyramidal geometry. In the crystal, inter\u00admolecular O\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network.In the title compound, {[NaTb(C DOI: 10.1107/S1600536810038456/is2603Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Cu(C7H4FO2)2(C10H14N2O)2(H2O)2], contains one-half of the mol\u00adecule. The CuII ion is located on an inversion centre, and is coordinated by two N atoms from two diethyl\u00adnicotinamide ligands, two O atoms from two 4-fluoro\u00adbenzoate (PFB) ligands and two water mol\u00adecules in a distorted octa\u00adhedral geometry. In the PFB ligand, the carboxyl\u00adate group is twisted at an angle of 2.10\u2005(14)\u00b0 from the attached benzene ring. In the crystal structure, inter\u00admolecular O\u2014H\u22efO hydrogen bonds link mol\u00adecules related by translation along the a axis into chains. Weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings of neighbouring mol\u00adecules [centroid-to-centroid distance = 3.571\u2005(2)\u2005\u00c5] further consolidate the crystal packing.The asymmetric unit of the title mononuclear Cu N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C10H14N2O)2(H2O)2] = 0.033 wR(F 2) = 0.099 S = 1.15 4109 reflections233 parameters2 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.57 e \u00c5\u22123 \u0394\u03c1min = \u22120.45 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811029941/cv6646Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, C\u2014H\u22ef\u03c0 inter\u00adactions [C\u2014H\u22efcentroid distances = 3.443\u2005(3) and 3.788\u2005(3)\u2005\u00c5] and N\u2014H\u22efS hydrogen bonds form layers parallel to (100). An intra\u00admolecular N\u2014H\u22efI hydrogen bond is also observed.In the mononuclear title complex, [CuI(C DOI: Click here for additional data file.10.1107/S1600536812044066/zb2025Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Each CuII atom is four-coordinated by two phenolate O and two imine N atoms from two symmetry-related Schiff base 2-{[2-(isopropyl\u00adaza\u00adnium\u00adyl)eth\u00adyl]imino\u00admeth\u00adyl}-6-meth\u00adoxy\u00adphenolate (L) ligands in a distorted square-planar geometry. The ammonium groups are involved in the formation of N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, which link one dication and two anions into an electroneutral cluster. When very weak Cu\u2014N interactions with a distance of 2.910\u2005(5)\u2005\u00c5 between the metal and the thiocyanate anions in apical positions are considered, the secondary coordination polyhedron is a very elongated CuN4O2 octahedron. The asymmetric unit of the title compound, [Cu(C For rel al. 2010; Cai 2](NCS)2 = 0.078 wR(F 2) = 0.143 S = 1.15 2406 reflections190 parameters6 restraintsH-atom parameters constrainedmax = 0.38 e \u00c5\u22123 \u0394\u03c1min = \u22120.36 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008SHELXTL (Sheldrick, 2008SHELXL97.Data collection: 10.1107/S1600536811001322/cv5037sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536811001322/cv5037Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "As a consequence, the Cl and Br atoms are not well ordered but distributed over the three possible positions such that the overall stiochiometry is two Br atoms and one Cl atom. The scrambling of the Br and Cl atoms indicates a small energy barrier for the exchange process between the apical and equatorial positions. Overall, the Te atom geometry is slightly distorted square pyramidal (\u03c4 = 0.052 for the major component). However, there is a weak secondary inter\u00adaction between the Te atoms and the disordered Br/Cl atoms of a nearby mol\u00adecule. The Te\u2014Br and Te\u2014Cl distances in both disorder components fall into two groups; a longer distance for the Br/Cl involved in this secondary inter\u00adaction [2.6945\u2005(17)\u2005\u00c5 for Br and 2.601\u2005(9)\u00c5 for Cl] and shorter bond distances to the remaining halogen atoms, indicating that this inter\u00adaction has slightly weakened the Te\u2014X bond, as is the case in the previously reported tribromido structure [Singh et al. (1990). J. Chem. Soc. Dalton Trans. pp. 907\u2013913]. Otherwise, the metrical parameters in the two structures are not significantly different. An intermolecular C\u2014H\u22efBr interaction occurs.The title compound, C \u00c5 b = 12.4785 (5) \u00c5 c = 14.4098 (6) \u00c5 \u03b2 = 98.200 (4)\u00b0V = 1296.61 (9) \u00c53 Z = 4 K\u03b1 radiationMo \u22121 \u03bc = 8.63 mmT = 123 K 0.63 \u00d7 0.50 \u00d7 0.10 mm Oxford Diffraction Xcalibur Ruby Gemini diffractometerCrysAlis PRO 2981 reflections with R int = 0.044 R[F 2 > 2\u03c3(F 2)] = 0.038 wR(F 2) = 0.056 S = 0.96 4241 reflections141 parameters1 restraintH-atom parameters constrainedmax = 0.91 e \u00c5\u22123 \u0394\u03c1min = \u22120.92 e \u00c5\u22123 \u0394\u03c1 CrysAlis PRO used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811054560/jj2114Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The TiIV atom displays a distorted tetra\u00adhedral geometry, with Ti\u2014O bond lengths ranging from 1.805\u2005(3) to 1.830\u2005(3)\u2005\u00c5 and O\u2014Ti\u2014O ligand bite angles of 100.16\u2005(12) and 101.36\u2005(12)\u00b0. The short Ti\u2014N bond distances, ranging from 1.877\u2005(4) to 1.905\u2005(4)\u2005\u00c5, indicate strong bonding between the TiIV atom and the dimethyl\u00adamide ligands.In the title four-coordinate complex, [Ti(C DOI: 10.1107/S1600536812002929/yk2040Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Pd\u2014P and Pd\u2014Cl bond lengths are 2.3550\u2005(7) and 2.2906\u2005(7)\u2005\u00c5, respectively. Some weak inter\u00adactions are observed between the aromatic rings of adjacent mol\u00adecules, with an inter\u00adplanar distance between two \u03c0-stacked rings of 3.505\u2005(3)\u2005\u00c5. Intra- and intermolecular C\u2014H\u22efCl hydrogen bonds also occur.The title compound, DOI: 10.1107/S1600536810042595/pk2275Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The oral administration of vitamin B12 offers a potentially simpler and cheaper alternative to parenteral administration, but its effectiveness has not been definitively demonstrated. The following protocol was designed to compare the effectiveness of orally and intramuscularly administered vitamin B12 in the treatment of patients \u00e2\u2030\u00a565\u00e2\u20ac\u2030years of age with vitamin B12 deficiency.The proposed study involves a controlled, randomised, multicentre, parallel, non-inferiority clinical trial lasting one year, involving 23 primary healthcare centres in the Madrid region (Spain), and patients \u00e2\u2030\u00a565\u00e2\u20ac\u2030years of age. The minimum number of patients required for the study was calculated as 320 (160 in each arm). Bearing in mind an estimated 8-10% prevalence of vitamin B12 deficiency among the population of this age group, an initial sample of 3556 patients will need to be recruited.Eligible patients will be randomly assigned to one of the two treatment arms. In the intramuscular treatment arm, vitamin B12 will be administered as follows: 1\u00e2\u20ac\u2030mg on alternate days in weeks 1 and 2, 1\u00e2\u20ac\u2030mg/week in weeks 3\u00e2\u20ac\u201c8,and 1\u00e2\u20ac\u2030mg/month in weeks 9\u00e2\u20ac\u201c52. In the oral arm, the vitamin will be administered as: 1\u00e2\u20ac\u2030mg/day in weeks 1\u00e2\u20ac\u201c8 and 1\u00e2\u20ac\u2030mg/week in weeks 9\u00e2\u20ac\u201c52. The main outcome variable to be monitored in both treatment arms is the normalisation of the serum vitamin B12 concentration at weeks 8, 26 and 52; the secondary outcome variables include the serum concentration of vitamin B12 (in pg/ml), adherence to treatment, quality of life (EuroQoL-5D questionnaire), patient 3satisfaction and patient preferences. All statistical tests will be performed with intention to treat and per protocol. Logistic regression with random effects will be used to adjust for prognostic factors. Confounding factors or factors that might alter the effect recorded will be taken into account in analyses.The results of this study should help establish, taking quality of life into account, whether the oral administration of vitamin B12 is an effective alternative to its intramuscular administration. If this administration route is effective, it should provide a cheaper means of treating vitamin B12 deficiency while inducing fewer adverse effects. Having such an alternative would also allow patient preferences to be taken into consideration at the time of prescribing treatment.This trial has been registered with ClinicalTrials.gov, number NCT 01476007, and under EUDRACT number 2010-024129-20. Vitamin B12 , along with other derivatives of folic acid, is a nutrient essential for the synthesis of DNA. Its deficiency is manifested through changes in the number and morphology of erythrocytes, leucocytes and platelets, and by neurological alterations owed to the progressive demineralisation of the nervous system (a consequence of defective myelin synthesis). Vitamin B12 is found mostly in food of animal origin. It is separated from ingested food through the action of the gastric acid, and in the duodenum the vast majority binds to intrinsic factor (IF). The vitamin B12/IF complex formed, which is very resistant to digestion, is then absorbed by endocytosis in the terminal ileum. Only 1-2% of vitamin B12 absorption occurs independent of IF .In the primary healthcare setting, the most commonly seen causes of vitamin B12 deficiency are related to abnormalities of digestion or absorption , and, to a lesser extent, a lack of exogenous supply. The exact prevalence of vitamin B12 deficiency in industrialised countries is unknown; indeed, different studies using different definitions have reported it as between 5% and 60% . ResultsIn the elderly, the symptoms of vitamin B12 deficiency caused by deficient diets and/or digestive and/or absorption problems can be nonspecific, making a diagnosis of deficiency more difficult. For example, up to 40% of elderly people show no haematological alterations. Further, neurological symptoms may appear before those of anaemia; indeed, only about 60% of elderly people with vitamin B12 deficiency are anaemic .The trial protocol was approved by the Madrid Region Clinical Research Ethics Committee in the last five years for vitamin B12 deficiency\u00c2\u00b7 serious neurological or psychiatric symptoms, including psychotic problems\u00c2\u00b7 dementia preventing the giving of informed consent to take part\u00c2\u00b7 atrophy of the optic nerve\u00c2\u00b7 serum folic acid concentration of <2.3\u00e2\u20ac\u2030ng/ml\u00c2\u00b7 stage 4 kidney disease 4 (estimated glomerular filtration rate [GFR] 15\u00e2\u20ac\u201c29\u00e2\u20ac\u2030ml/min)\u00c2\u00b7 having received/suffering malabsorption-related:\u00e2\u2014\u2039 surgery or diseases affecting the jejunum-ileum\u00e2\u2014\u2039 inflammatory-intestinal disease, e.g., Crohn\u00e2\u20ac\u2122s disease, ulcerative colitis\u00e2\u2014\u2039 celiac disease\u00c2\u00b7 chronic pancreatitis\u00c2\u00b7 myelodisplasia or malignant blood disease\u00c2\u00b7 haemophilia or other coagulation problems contraindicating parenteral administration\u00c2\u00b7 severe systemic disease\u00c2\u00b7 having been involved in any other trial involving the administration of any experimental treatment in the 28\u00e2\u20ac\u2030days prior to the start of the present study\u00c2\u00b7 being treated for HIV, HVB or HVC infection\u00c2\u00b7 hypersensitivity to vitamin B12, or any of the vitamin preparation\u00e2\u20ac\u2122s excipients\u00c2\u00b7 receiving anticoagulation treatment\u00c2\u00b7 being away from home and with no intention of residing for the following year in the health district where consultation was made\u00c2\u00b7 failing to meet any inclusion criterion\u00c2\u00b7 limitations regarding oral treatmentParticipants will be enrolled consecutively by their general practitioners when attending a primary healthcare centre in the study area Figure . All patThe sample size required was determined bearing in mind the results of Kuzminski et al. . In the Assuming that the percentage of patients showing an increase in serum vitamin B12 concentration to above 179\u00e2\u20ac\u2030pg/ml in both groups is 70%, means the study requires at least 304 patients (152 in each arm) for a threshold of non-inferiority of 10% and a statistical power of 60% with significance set at p\u00e2\u20ac\u2030<\u00e2\u20ac\u20300.05. Given the type of patients to be studied, i.e., patients who have come to the health centres for consultation, plus the fact that their own family doctors are members of the research team, a loss to follow-up of under 5% is expected. The minimum starting sample size for each arm was therefore deemed to be n\u00e2\u20ac\u2030=\u00e2\u20ac\u2030160. With an expected prevalence of vitamin B12 deficiency of 8-10% , the original number of patients to be enrolled so that 320 with a vitamin B12 deficiency can be guaranteed is 3556.In studies with the present design it is impossible to blind the patient to the treatment received. However, this limitation is compensated for by the objective measurement of the main outcome variable (the serum vitamin B12 concentration) and the randomisation of the patients to the treatment groups. Further, the persons charged with the statistical analysis of the data will be blind to the identity of the patients in each treatment arm.The pharmaceutical formulations to be used in the study are commercially available in Spain. The treatments will involve:\u00c2\u00b7 Intramuscular route: 1\u00e2\u20ac\u2030mg of vitamin B12 on alternate days during weeks 1 and 2; 1\u00e2\u20ac\u2030mg/week over weeks 3\u00e2\u20ac\u201c8 ; and 1\u00e2\u20ac\u2030mg/month from weeks 9\u00e2\u20ac\u201c52\u00c2\u00b7 Oral route: 1\u00e2\u20ac\u2030mg/day of vitamin B12 for 8\u00e2\u20ac\u2030weeks; 1\u00e2\u20ac\u2030mg/week from weeks 9\u00e2\u20ac\u201c52Patients in both arms will undergo analytical monitoring in weeks 8, 26 and 52. They will receive appointments for the appropriate dates. The response to treatment will be recorded alongside adherence to treatment and the appearance of any adverse effects.Before work begins, the project will be presented to all the research team members in a special meeting. Training sessions lasting 2\u00e2\u20ac\u201c3\u00e2\u20ac\u2030h will also be held at each participating health centre. These will involve a review of the inclusion and exclusion criteria, provide instructions regarding the intervention, and examine the ethical requirements to be met for the trial to be held.The procedures to be followed and information to be recorded at each of a patient\u00e2\u20ac\u2122s visits to a participating health centre is as follows:Selection Visit\u00c2\u00b7 Signing of informed consentAssessment of inclusion/exclusion criteriaRecording of demographic data (age and sex)Analysis: serum vitamin B12. If concentration is <179 pg/ml the following analyses are to be requested: haemogram, biochemical analysis , ferritin, folic acid, anti-IF antibody level. If serum vitamin B12 concentration is >179 pg/ml: patient preference questionnaireRandomisation of patients to treatment groupVisit 1 (start of treatment)\u00c2\u00b7 Anamnesis: record whether the patient lives alone or with others, lifestyle habits, use of alcohol, whether a vegan diet is followed, whether the patient has undergone gastrectomySymptoms: record paresthesia, asthenia, loss or reduction of appetite, sadness or change in state of mind, concomitant pharmacological treatmentPhysical examination: for Hunter\u00e2\u20ac\u2122s glositis, positional and vibrational sensitivityQuestionnaires: Lobo cognitive mini-exam, EuroQoL-5DRecord concomitant treatmentRequest analyses to be performed one week before next visit: haemogram and serum vitamin B12Therapeutic plan: patient in oral arm \u00e2\u20ac\u201c provision of medication; patient in intramuscular arm \u00e2\u20ac\u201c provide appointments for injectionsVisit 2 (week 8)\u00c2\u00b7 Anamnesis: record lifestyle habits and use of alcoholSymptoms: if pathological at the first visit, record paresthesia, asthenia, loss or reduction of appetite, sadness or change in level of happiness, and concomitant pharmacological treatmentPhysical examination: if pathological at the first visit examine for Hunter\u00e2\u20ac\u2122s glositis, positional and vibrational sensitivityRecord concomitant treatmentRequest analyses to be performed one week before next visit: haemogram and serum vitamin B12Questionnaires: EuroQoL-5DAssessment of adverse effectsTherapeutic plan: patient in oral arm \u00e2\u20ac\u201c provision of medication; patient in intramuscular arm \u00e2\u20ac\u201c provide appointments for injectionsAssess adherence to treatment: oral route \u00e2\u20ac\u201c count number of vials used; intramuscular route: count injections givenVisit 3 (week 26)\u00c2\u00b7 Anamnesis: record lifestyle habits and use of alcoholSymptoms: if pathological at the first visit, record paresthesia, asthenia, loss or reduction of appetite, sadness or change in level of happiness, and concomitant pharmacological treatmentPhysical examination: if pathological at the first visit examine for Hunter\u00e2\u20ac\u2122s glositis, positional and vibrational sensitivityRecord concomitant treatmentRequest analyses to be performed one week before next visit: haemogram and serum vitamin B12Questionnaire: EuroQoL-5DAssessment of adverse effectsTherapeutic plan: patient in oral arm \u00e2\u20ac\u201c provision of medication; patient in intramuscular arm \u00e2\u20ac\u201c provide appointments for injectionsAssess adherence to treatment: oral route \u00e2\u20ac\u201c count number of vials used; intramuscular route: count injections givenVisit 4 (week 52)\u00c2\u00b7 Anamnesis: record lifestyle habits and use of alcoholSymptoms: record paresthesia, asthenia, loss or reduction of appetite, sadness or change in level of happiness, and concomitant pharmacological treatmentPhysical examination: for Hunter\u00e2\u20ac\u2122s glositis, positional and vibrational sensitivityRecord concomitant treatmentQuestionnaires: EuroQoL-5D, satisfaction and preferencesAssessment of haemogram and serum vitamin B12 concentrationAssessment of adverse effectsAssess adherence to treatment: oral route \u00e2\u20ac\u201c count number of vials used; intramuscular route: count injections givenThe main outcome to be measured is the normalisation of the serum vitamin B12 concentration (>179\u00e2\u20ac\u2030pg/ml) at 8, 26 and 52\u00e2\u20ac\u2030weeks. The secondary outcomes will be the serum vitamin B12 concentration (pg/ml), adverse events , adherence to treatment , quality of life (measured using the EuroQoL-5D questionnaire), and patient satisfaction and preferences.Including age, sex, whether the patient lives alone or with others, whether a vegan diet is followed, and the use of alcohol (g/week).Symptoms such as paresthesia, asthenia, loss or reduction of appetite, sadness or change in state of mind (anamnesis), Hunter\u00e2\u20ac\u2122s glositis, positional and vibrational sensitivity , and cognitive decline (Lobo test).Haemogram (complete blood cell and platelet count) and biochemical analysis . Blood analyses will be performed in plasma or serum as required and under standard conditions.Recording of the taking of protein pump inhibitors, H2 receptor antagonists, antacids, potassium, metformin, colchicine, neomycin, p-aminosalicylic acid, parenteral chloramphenicol, Fe, vitamin C and other vitamin supplements.Patients will be removed from the trial if any of the following conditions are met:\u00c2\u00b7 Serum vitamin B12 concentration still <179\u00e2\u20ac\u2030pg/ml after 8\u00e2\u20ac\u2030weeks of treatment. Treatment will be deemed to have failed in these patients, and they will be further studied and treated outside the trial according to normal clinical practice.\u00c2\u00b7 Serious adverse events.\u00c2\u00b7 Voluntary withdrawal or violation of the protocol.At least two attempts will be made to contact by telephone those patients who do not come for their scheduled visits. All patients will be informed that they can abandon the study at any time without this affecting their future medical treatment in any way.The trial will involve a descriptive statistical analysis of the baseline characteristics of patients in both treatment arms. Quantitative variables will be described in terms of their measure of central tendency, mean or median (for those showing asymmetric distributions), and the corresponding dispersion, standard deviation or interquartile range. Qualitative variables will be described in terms of proportions and their corresponding confidence intervals.The Student t test or Mann\u00e2\u20ac\u201cWhitney U test will be used to determine whether the two treatment arms are comparable based on their quantitative baseline characteristics and known prognostic factors. Comparisons on qualitative variables will be undertaken using the Pearson Chi-squared test or Fisher\u00e2\u20ac\u2122s Exact test as required. If cases of inequality are detected, the confounding factors will be defined and appropriate adjustments made.Intention-to-treat and per-protocol analyses will both be performed, as is recommended for non-inferiority studies .The effectiveness of treatment will be analysed by examining the therapeutic success achieved in each arm at 8, 26 and 52\u00e2\u20ac\u2030weeks, determining the 95% confidence interval for the percentage of patients in each treatment arm whose serum vitamin B12 concentrations become normalised. If the confidence intervals do not fall outside the non-inferiority limit (10%), it can be concluded that the oral treatment is not inferior to the intramuscular treatment. The within-patient percentage change in serum vitamin B12 concentration at each monitoring point will be determined, and the confidence intervals for the difference in the mean values for each arm calculated.If the distribution of confounding factors differs in the two arms, explicative regression analysis will be performed in which the dependent variable will be the normalisation of the serum vitamin B12 concentration, and the independent variable will be the treatment group.Repeated measures ANOVA will be used to examine the change in serum vitamin B12 concentration in each group at each monitoring point.The incidence of adverse events in the two arms will be compared using the Pearson Chi-squared test or Fisher\u00e2\u20ac\u2122s Exact test as required.-based quality of life values.The perception of quality of life by the patients of each arm will be assessed by comparing the EuroQol 5D scores and the transformation of these scores into utilityAdherence to treatment will be examined via the counting of oral doses taken in the oral arm, and the number of injections given in the intramuscular arm. An operative indicator variable will then be defined to describe the degree of adherence.th 2011). It will be performed by qualified medical and scientific staff. The rights and welfare of the patients will be respected at all times. All patients will be adequately informed, both verbally and in writing, of the nature of the trial, its aim, and its risks and possible benefits. Given that the study is a non-inferiority trial, all patients will be informed that the oral treatment is expected to be as effective as the standard intramuscular treatment. Signed, dated consent to be included will be required from each patient.The trial has been approved by the Madrid Region Clinical Research Ethics Committee (February 8Convenio de Oviedo) (1997).Spanish law regarding the use of human subjects in clinical trials will be adhered to. The trial will respect all basic ethical principles of autonomy, justice, goodness of intent and absence of malintent according to the standards of good clinical practice enshrined in the Declaration of Helsinki and the Oviedo Agreement that quality scientific information supports the effectiveness of the therapeutic options on offer, and 2) that heterogeneous groups of patients have recorded their satisfaction with these options. The present trial provides for information in this respect to be gathered and therThe trial is also designed to provide information on the effect of the normalisation of serum vitamin B12 concentrations by both treatments on patient-perceived quality of life. Physicians commonly assume that taking oral supplements will be associated with a feeling of greater well-being, although this has never been proven . The preThe trial suffers from the practical limitation of having to enrol a large number of patients to meet its sample size requirements. However, a high degree of motivation is expected of the research team since its clinical assistance members are those involved in the enrolment process. Further, the fact that the patients to be enrolled will be seeking medical help suggests few will be lost to follow-up. A further possible limitation is the low statistical power used in the calculation of the sample size. The 60% power contemplated requires a sample size of 304 patients (152 in each arm) \u00e2\u20ac\u201c higher powers would increase the sample size required and the enrolment of such numbers cannot be guaranteed. However, given the results reported in previous studies that used moderate/high doses of vitamin B12, it should be possible to demonstrate the non-inferiority of the oral treatment with this power level. If the 95% confidence interval were to cross the non-inferiority threshold, i.e., showing the results to be inconclusive, the intramuscular treatment would remain the treatment of choice. To determine the degree of adherence to treatment (and thus avoid outcome dilution effects) , the numThe decision not to take serum methylnalonic acid and homocysteine concentrations into account as diagnostic markers and outcome variables was made bearing in mind that these are not normally determined, either at diagnosis or during follow-up, in patients with a vitamin B12 deficiency.Finally, given the pragmatic nature of the proposed trial, the decision was taken to include consecutive patients seeking medical help at the participating centres, thus ensuring the enrolment of subjects similar to those that would be seen in normal clinical practice.Fe: Ferrum; g: Gram; GFR: Glomerular filtration rate; GGT: Gamma-glutamyl transpeptidase; GOT: Glutamic oxaloacetic transaminase; GP: General practitioner; GPT: Glutamic-pyruvic transaminase; HIV: Human immunodeficiency virus; HVB: Hepatitis B virus; HVC: Hepatitis C virus; IF: Intrinsic factor; \u00ce\u00bcg: Microgram; MMA: Methylmalonic acid; mg: Milligrams; ng: Nanograms; pg: Picograms.The authors declare that they have no competing interests.PGE y RRF conceived of the study and participated in its design. TSC; RRF; SGE; IdCG; JMF; EEM; participated in the design and coordination of the study. FRS; MGS; RGG; MAMS; COL; MLSP; CMR; BMB; AVP; FGBG; JEMS; RRB; GAC; LMCB; EPC; MRB; MTRM; SSD; SMI; RRG; IBL; MVN; JSD; TGG; MDC; AAB participated in different phases of the design. TSC; RRF; SGE; IdCG; JMF; EEM directed the writing of the manuscript. All authors OB12 Group read and approved the final manuscript.Healthcare Centre (HC) Barajasx: Germ\u00c3\u00a1n Reviriego Ja\u00c3\u00a9n, Cristina Montero Garc\u00c3\u00ada, Ana Isabel Sanz Lorente, Ma del Pilar Serrano Simarro, Juli\u00c3\u00a1n D\u00c3\u00adaz S\u00c3\u00a1nchez, Irma Ma Ramos Guti\u00c3\u00a9rrez, Josefa Ma San Vicente Rodr\u00c3\u00adguez, Pilar Huelin Mart\u00c3\u00adn, Ma Inmaculada Gonz\u00c3\u00a1lez Garc\u00c3\u00ada, Margarita Camarero Shelly, Clarisa Reinares Mart\u00c3\u00adnez, Laura Villanova Cuadra, Rosa Ma\u00e2\u20ac\u2030G\u00c3\u00b3mez del Forcallo. HC Doctor Cirajas: Francisco Endrino G\u00c3\u00b3mez, Ma Rosario Ferreras Eleta, Luis De Vicente Aymat, Mar\u00c3\u00ada Santos Santander Guti\u00c3\u00a9rrez, Alicia Mateo Madurga. HC Juncal: Nuria Caballero Ram\u00c3\u00adrez, Ana Mor\u00c3\u00a1n Escudero, Mercedes Rodr\u00c3\u00adguez Franco, M\u00c2\u00aa Luz Meiri\u00c3\u00b1o P\u00c3\u00a9rez, M\u00c2\u00aa Mar Zamora G\u00c3\u00b3mez, Francisco Vivas Rubio, Mar\u00c3\u00ada Mart\u00c3\u00adn Mart\u00c3\u00adn. HC Miguel de Cervantes: Rafael P\u00c3\u00a9rez Quero, M\u00c2\u00aa Isabel Manzano Mart\u00c3\u00adn, Raimundo Pastor S\u00c3\u00a1nchez, Alicia Herrero de Dios, Cesar Redondo Luci\u00c3\u00a1\u00c3\u00b1ez. HC Reyes Magos: Cristina Casado Rodr\u00c3\u00adguez, Luisa Mar\u00c3\u00ada Andr\u00c3\u00a9s Arreaza, Pilar Hombrados Gonzalo, Soledad Escolar Llamazares, Francisco L\u00c3\u00b3pez Ortiz, Luz M\u00c2\u00aa del Rey Moya, Isabel Rodr\u00c3\u00adguez L\u00c3\u00b3pez. HC Calesas: Diego Mart\u00c3\u00adn Acicoya, Pilar Kloppe Villegas, Isabel Garc\u00c3\u00ada Amor, Magdalena Canals Aracil, Jos\u00c3\u00a9 Javier G\u00c3\u00b3mez Marco, Alberto Gonz\u00c3\u00a1lez \u00c3\ufffdlvaro, Fco Javier San Andr\u00c3\u00a9s Rebollo, In\u00c3\u00a9s Gonz\u00c3\u00a1lez L\u00c3\u00b3pez, Isabel Herreros Hernanz, Antonio Revuelta Alonso, Nieves Calvo Arrabal, M\u00c2\u00aa Milagros Jimeno Gal\u00c3\u00a1n, Rosa Garc\u00c3\u00ada Hern\u00c3\u00a1ndez. HC Guayaba: Tom\u00c3\u00a1s G\u00c3\u00b3mez Gasc\u00c3\u00b3n, Concepci\u00c3\u00b3n Vargas-Machuca Caba\u00c3\u00b1ero, M\u00c2\u00aa Isabel Guti\u00c3\u00a9rrez S\u00c3\u00a1nchez, M\u00c2\u00aa Angeles Fern\u00c3\u00a1ndez Abad, Margarita Beltejar Rodr\u00c3\u00adguez, Javier Mart\u00c3\u00adnez Suberviola, Miguel Angel Real P\u00c3\u00a9rez, Carmen Coello Alarc\u00c3\u00b3n, Carlos San Andr\u00c3\u00a9s Pascua, Jos\u00c3\u00a9 Antonio Granados Garrido. HC General Ricardos: Santiago Mach\u00c3\u00adn Hamalainen, Raquel Mateo Fern\u00c3\u00a1ndez, Cristina de la C\u00c3\u00a1mara Gonzalez, Jos\u00c3\u00a9 D.Garc\u00c3\u00a9s Ranz, Asunci\u00c3\u00b3n Prieto Orzanco, M\u00c2\u00aa Teresa Mar\u00c3\u00adn Becerra, Paulino Cubero Gonz\u00c3\u00a1lez, Francisco R. Abell\u00c3\u00a1n L\u00c3\u00b3pez, Olga \u00c3\ufffdlvarez Montes, Mercedes Canellas Manrique, M\u00c2\u00aa Jos\u00c3\u00a9 San Telesforo Navarro, M\u00c2\u00aa Mercedes Parrilla Laso, M\u00c2\u00aa \u00c3\ufffdngeles Aragoneses Ca\u00c3\u00b1as, Angela Au\u00c3\u00b1\u00c3\u00b3n Muelas HC Los Y\u00c3\u00a9benes, Esther Vald\u00c3\u00a9s Cruz, Consuelo Mayoral Lopez, Teresa Gijon Seco, Francisca Martinez Vallejo. HC Valle Incl\u00c3\u00a1n: Ana Isabel Men\u00c3\u00a9ndez Fern\u00c3\u00a1ndez, M\u00c2\u00aa del Mar De la Pe\u00c3\u00b1a Gonz\u00c3\u00a1lez, M\u00c2\u00aa \u00c3\ufffdngeles Maroto Garc\u00c3\u00ada, Mar\u00c3\u00ada S\u00c3\u00a1nchez Crist\u00c3\u00b3bal. HC Lavapi\u00c3\u00a9s: M\u00c2\u00aa Carmen \u00c3\ufffdlvarez Orviz, Jes\u00c3\u00bas Herrero Hern\u00c3\u00a1ndez, M\u00c2\u00aa Veredas Gonz\u00c3\u00a1lez M\u00c3\u00a1rquez, M\u00c2\u00aa Jes\u00c3\u00bas L\u00c3\u00b3pez Rodr\u00c3\u00adguez, M\u00c2\u00aa de las Maravillas Almarza Garc\u00c3\u00ada, M\u00c2\u00aa Teresa San Clemente Pastor, M\u00c2\u00aa \u00c3\ufffdmparo Corral Rubio. HC Colmenar Viejo Norte: Gonzalo Ruiz Zurita, \u00c3\ufffdngela Allue Bergua, Marta Cabrera Orozco, M\u00c2\u00aa del Puerto De Antonio Garc\u00c3\u00ada, Ana Isabel Cerezo Diviu, Inmaculada Solsons Roig, Pilar G\u00c3\u00b3mez de Abia. HC Fuentelarreina: Mar\u00c3\u00ada Concepci\u00c3\u00b3n D\u00c3\u00adaz Laso, M\u00c2\u00aa Luisa Asensio Ruiz, Carmen Siguero P\u00c3\u00a9rez. HC Presentaci\u00c3\u00b3n Sabio: Antonio Molina Siguero, Inmaculada Cerrada Puri, Paloma Rodr\u00c3\u00adguez Almagro, Rosa Rosanes Gonz\u00c3\u00a1lez, M\u00c2\u00aa Carmen P\u00c3\u00a9rez Garc\u00c3\u00ada. HC Cuzco: Mar Noguerol \u00c3\ufffdlvarez, M\u00c2\u00aa \u00c3\ufffdngeles de Miguel Abanto, M\u00c2\u00aa Lourdes Reyes Mart\u00c3\u00adnez, Pilar Guti\u00c3\u00a9rrez Valent\u00c3\u00adn, Jorge G\u00c3\u00b3mez Ciriano, Raquel Calzada Benito, Carolina Torrijos Bravo, David Ferreiro Gonz\u00c3\u00a1lez, Judit Le\u00c3\u00b3n Gonz\u00c3\u00a1lez. HC San Mart\u00c3\u00adn de Valdeiglesias: Nuria Tom\u00c3\u00a1s Garc\u00c3\u00ada, Alberto Alcal\u00c3\u00a1 Fa\u00c3\u00bandez, Eva Fern\u00c3\u00a1ndez L\u00c3\u00b3pez, In\u00c3\u00a9s Melero Redondo, Ricardo Gonz\u00c3\u00a1lez Gasc\u00c3\u00b3n. HC Pedroches: Jeannet S\u00c3\u00a1nchez Y\u00c3\u00a9pez, Mercedes del Pilar Fern\u00c3\u00a1ndez Gir\u00c3\u00b3n, Beatriz L\u00c3\u00b3pez Serrano, M\u00c2\u00aa Teresa Rodr\u00c3\u00adguez Monje, Paloma Morso Pelaez, Mar\u00c3\u00ada Cortes Duran, Carolina L\u00c3\u00b3pez Olmeda, Almudena Garc\u00c3\u00ada- Uceda Sevilla, Dolores Serrano Gonz\u00c3\u00a1lez, Inmaculada Santamar\u00c3\u00ada L\u00c3\u00b3pez. HC Mendiguch\u00c3\u00ada Carriche: Francisca Garc\u00c3\u00ada De Blas Gonz\u00c3\u00a1lez, Alberto L\u00c3\u00b3pez Garc\u00c3\u00ada-Franco, Amaya Azcoaga Lorenzo, Mar \u00c3\ufffdlvarez Villalba, Bel\u00c3\u00a9n Pose Garc\u00c3\u00ada. HC Santa Isabel: Rosa Fern\u00c3\u00a1ndez Garc\u00c3\u00ada, Francisco de Alba G\u00c3\u00b3mez, Antonio Redondo Horcajo, Beatriz Pajuelo M\u00c3\u00a1rquez, Jos\u00c3\u00a9 Luis Gala Paniagua, Encarnaci\u00c3\u00b3n Cidoncha Calder\u00c3\u00b3n, \u00c3\ufffdngel Delgado Delgado, M\u00c2\u00aa Jes\u00c3\u00bas G\u00c3\u00b3mez Mart\u00c3\u00adn, Jos\u00c3\u00a9 Francisco \u00c3\ufffdvila Tomas. HC El Greco: Jos\u00c3\u00a9 Enrique Mari\u00c3\u00b1o Su\u00c3\u00a1rez, Jos\u00c3\u00a9 Luis Quintana G\u00c3\u00b3mez, Jos\u00c3\u00a9 Antonio Gonz\u00c3\u00a1lez-Posada Delgado, Enrique Revilla Pascual, Esperanza Duralde Rodr\u00c3\u00adguez, Milagros Beamud Lagos. HC Arroyo de la Media Legua: Leonor Gonz\u00c3\u00a1lez Gal\u00c3\u00a1n, Mar\u00c3\u00ada Verdugo Rosado, Luis Nistal Mart\u00c3\u00adn de Serranos, M\u00c2\u00aa Jes\u00c3\u00bas L\u00c3\u00b3pez Barroso, Mariano Rivera Moreno, Margarita Torres Parras, M\u00c2\u00aa Reyes Delgado Pulpon, Elena Alcal\u00c3\u00a1 Llorente. HC Federica Montseny: Sonsoles Mu\u00c3\u00b1oz Moreno, Ana Mar\u00c3\u00ada Ribao Verdugo, Mar\u00c3\u00ada Jes\u00c3\u00bas Fidalgo Baz, Isabel Vaquero Turi\u00c3\u00b1o, Ana Mar\u00c3\u00ada Je\u00c3\u00ba Fidalgo Baz, Clementa Sanz Sanchez, Ana Mar\u00c3\u00ada S\u00c3\u00a1nchez Sempere, Javier Mart\u00c3\u00adnez Sanz, Mar\u00c3\u00ada Isabel Arratibel Elizondo. HC Buenos Aires: Paloma Gonz\u00c3\u00a1lez Escobar, Javier Mu\u00c3\u00b1oz Guti\u00c3\u00a9rrez, Raquel Ba\u00c3\u00b1os Morras, Carmen Molins Santos, Ana Mar\u00c3\u00ada Ibarra S\u00c3\u00a1nchez, Cecilio G\u00c3\u00b3mez Almod\u00c3\u00b3var, Cristina Cassinello Espinosa.Ministerio de Sanidad, Pol\u00c3\u00adtica Social e Igualdad, Spain and by CAIBER - Spanish Clinical Research Network. The authors thank the following persons for their contributions to this work: Dolores Otero-Criado, Carlos Carvajales Fern\u00c3\u00a1ndez, Rosa Zurdo-Baz and Raisa Gonz\u00c3\u00a1lez-P\u00c3\u00a9rez.This study was funded by the The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/12/394/prepub"} +{"text": "The asymmetric unit further contains one 4-meth\u00adoxy\u00adbenzoate anion, one nicotinamide (NA) ligand and one coordinated and one uncoordinated water mol\u00adecule; all ligands are monodentate. The four O atoms in the equatorial plane around the NiII ion form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two pyridine N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the attached benzene ring is 7.2\u2005(1)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 72.80\u2005(4)\u00b0. An intra\u00admolecular O\u2014H\u22efO hydrogen bond links the uncoordinated water mol\u00adecule to one of the carboxyl\u00adate groups. In the crystal structure, inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network.In the mononuclear title compound, [Ni(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2]\u00b72H2O = 0.026 wR(F 2) = 0.067 S = 1.04 3740 reflections228 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.41 e \u00c5\u22123 \u0394\u03c1min = \u22120.32 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S1600536810025985/ci5126sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810025985/ci5126Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two pyridine rings in L form a dihedral angle of 50.0\u2005(2)\u00b0. L ligands bridge adjacent HgCl2 units into polymeric chains propagating in [010]. The crystal packing is further stabilized by weak inter\u00admolecular C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings, with a centroid\u2013centroid separation of 3.529\u2005(9)\u2005\u00c5.In the title coordination polymer, [HgCl DOI: 10.1107/S1600536812035775/cv5327Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The small deviations from the ideal tetra\u00adhedral geometry around the P atoms are illustrated by C\u2014P\u2014C angles ranging from 104.08\u2005(9) to 106.46\u2005(9)\u00b0. In the crystal, the mol\u00adecules are linked by weak C\u2014H\u22efF, C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, [Ir(C DOI: 10.1107/S1600536812035593/hb6932Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "IV atom in the centrosymmetric title complex, [Sn2(CH3)4(NCS)2(OH)2], adopts a distorted trigonal\u2013bipyramidal coordination environment defined by two methyl C atoms and one bridging hydroxide group in the equatorial plane while the other bridging hydroxide group and the N atom of the thio\u00adcyanate anion are in the apical >positions. The dinuclear species are linked through O\u2014H\u22efS and C\u2014H\u22ef S hydrogen-bonding inter\u00adactions into a three-dimensional network.The Sn For str al. 2007; Ng 4(NCS)2(OH)2] = 0.026wR(F2) = 0.056S = 1.111603 reflections71 parameters1 restraintH atoms treated by a mixture of independent and constrained refinementmax = 1.06 e \u00c5\u22123\u0394\u03c1min = \u22120.64 e \u00c5\u22123\u0394\u03c1COLLECT used to solve structure: SIR97 I, global. DOI: Click here for additional data file.10.1107/S1600536812043462/wm2689Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II ion in the title complex, [Pd(NCS)2(C14H10N4)], is four-coordinated in a distorted square-planar environment by the two pyridine N atoms of the chelating 2,3-di-2-pyridyl\u00adpyrazine (dpp) ligand and two S atoms from two thio\u00adcyanate anions. The pyridine rings are considerably inclined to the least-squares plane of the PdS2N2 unit [maximum deviation = 0.027\u2005(1)\u2005\u00c5], making dihedral angles of 70.3\u2005(2) and 69.2\u2005(1)\u00b0. The pyrazine ring is almost perpendicular to the PdS2N2 plane, with a dihedral angle of 86.3\u2005(1)\u00b0. The thio\u00adcyanate ligands are located on opposite sides of the PdS2N2 unit plane and are almost linear [N\u2014C\u2014S angles = 177.8\u2005(6) and 178.9\u2005(6)\u00b0]. The complex mol\u00adecules are stacked in columns along the b axis and are connected by inter\u00admolecular C\u2014H\u22efN hydrogen bonds, forming chains along the a axis.The Pd X2(dpp)] , see: Ha ] = 0.046wR(F2) = 0.100S = 1.003247 reflections226 parametersH-atom parameters constrainedmax = 0.89 e \u00c5\u22123\u0394\u03c1min = \u22120.66 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II complex, [Cd2(C7H5O4)4(C12H8N2)2(H2O)2], the CdII cation is coord\u00adinated by a bidentate phenanthroline (phen) ligand, three dihy\u00addroxy\u00adbenzoate (dhba) anions and one water mol\u00adecule in a distorted CdN2O4 octa\u00adhedral geometry. Among the dhba anions, two anions bridge two CdII cations to form the dimeric complex with significant different Cd\u2014O bond distances of 2.2215\u2005(19) and 2.406\u2005(2)\u2005\u00c5. The centroid\u2013centroid distance of 3.4615\u2005(19)\u2005\u00c5 between two nearly parallel benzene rings of the dhba and phen ligands coordinating to the same CdII cation indicates the existence of intra\u00admolecular \u03c0\u2013\u03c0 stacking in the complex. Extensive O\u2014H\u22efO hydrogen bonding and inter\u00admolecular weak C\u2014H\u22efO hydrogen bonding help to stabilize the crystal structure. One hy\u00addroxy group of the monodentate dhba ligand is disordered over two sites with a site-occupancy ratio of 0.9:0.1.In the title centrosymmetric dimeric Cd DOI: 10.1107/S1600536810021252/ng2775Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The \u2013C\u00a0CH and the chlorine ligands are disordered over two equivalent positions (0.5 occupancy each). The coordination geometry is distorted octa\u00adhedral, with the \u2013C\u00a0CH fragment and the Cl ligand in trans positions. The four P atoms occupy the equatorial plane of the octa\u00adhedron and the chloride and acetyl\u00adide ligands the axial positions.The mol\u00adecular structure of the title compound, Cl(C26H24P2)2] = 0.030wR(F2) = 0.072S = 1.096470 reflections293 parametersH-atom parameters constrainedmax = 0.49 e \u00c5\u22123\u0394\u03c1min = \u22120.48 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 global, I. DOI: Click here for additional data file.10.1107/S1600536812044558/bg2483Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The asymmetric unit contains two complex cations, two hexa\u00adfluoridophosphate anions and one uncoordinated water mol\u00adecule. In one of the hexa\u00adfluoridophosphate anions, four of the F aroms are disordered over two sets of sites in a 0.632\u2005(11):0.368\u2005(11) ratio. In the crystal, the cations, anions and water mol\u00adecules are connected by O\u2014H\u22efO and O\u2014H\u22efF hydrogen bonds. \u03c0\u2013\u03c0 inter\u00adactions are present between the pyridine rings [centroid\u2013centroid distance = 3.814\u2005(1)\u2005\u00c5].In the title complex, [Co(C DOI: 10.1107/S1600536811023270/hy2438Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal packing exhibits weak inter\u00admolecular Cl\u22efCl contacts of 3.279\u2005(4)\u2005\u00c5, \u03c0\u2013\u03c0 inter\u00adactions between the substituted Cp rings of two neighbouring 2-chloro-1-ferrocenyl\u00adethanone mol\u00adecules [centroid\u2013centroid distance = 3.534\u2005(3)\u2005\u00c5], and weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds. In the title co-crystal, [Fe(C DOI: 10.1107/S1600536811038244/cv5144Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each paddle-wheel unit is joined to four such neighbours through bridging dicarboxyl\u00adate ligands, producing a two-dimensional undulating layer parallel to (-101). Adjacent sheets are stacked in a parallel fashion to form a three-dimensional supra\u00admolecular structure which is stabilized by inter\u00adlayer \u03c0\u2013\u03c0 inter\u00adactions between benzene rings, with a centroid\u2013centroid distance of 3.722\u2005\u00c5. The range of Zn\u2014O bond lengths is 2.0440\u2005(17)\u20132.1256\u2005(15)\u2005\u00c5 and the Zn\u2014Cl bond length is 2.2622\u2005(6)\u2005\u00c5.The title compound, [Zn(C Cl] = 0.027wR(F2) = 0.078S = 1.072640 reflections181 parametersH-atom parameters constrainedmax = 0.45 e \u00c5\u22123\u0394\u03c1min = \u22120.45 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812020077/bg2452Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The thiophene rings form different dihedral angles [40.15\u2005(9) and 15.43\u2005(10)\u00b0] with the pyrazine ring. A strong \u03c0\u2013\u03c0 stacking inter\u00adaction occurs between adjacent pyrazine\u00adphenanthroline units with an inter\u00adplanar distance of 3.4352\u2005(16)\u2005\u00c5.The mol\u00adecule of the title compound, C DOI: 10.1107/S160053681201522X/aa2049Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The MnII atom is coordinated by four N atoms [Mn\u2014N = 2.2168\u2005(14) and 2.2407\u2005(14)\u2005\u00c5] from two 2,2\u2032-biimidazole ligands and two water mol\u00adecules [Mn\u2014O = 2.2521\u2005(14)\u2005\u00c5] in a distorted octa\u00adhedral geometry. Inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds consol\u00adidate the crystal packing, which also exhibits \u03c0\u2013\u03c0 inter\u00adactions between five-membered rings, with a centroid\u2013centroid distance of 3.409\u2005(2)\u2005\u00c5.The asymmetric unit of the title compound, [Mn(C DOI: 10.1107/S160053681202199X/cv5293Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The 2,2\u2032-bipyridine mol\u00adecule is slightly twisted with a dihedral angle between the pyridine rings of 7.35\u2005(12)\u00b0. Intra\u00admolecular C\u2014H\u22efS hydrogen bonds are present. In the crystal, mol\u00adecules are stacked along the c axis by \u03c0\u2013\u03c0 inter\u00adactions, with centroid\u2013centroid distances of 3.6021\u2005(15) and 3.6401\u2005(16)\u2005\u00c5. The crystal structure also features weak C\u2014H\u22ef\u03c0 inter\u00adactions.In the title benzene-solvated heteroleptic lithium complex, [Li(C DOI: Click here for additional data file.10.1107/S160053681300456X/rz5044Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CuII atoms are linked by the bridging ligands into a layer parallel to (101). The presence of intra\u00adlayer O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the pyridine and benzene rings [centroid\u2013centroid distances = 3.808\u2005(2) and 3.927\u2005(2)\u2005\u00c5] stabilizes the layer. Further O\u2014H\u22efO hydrogen bonds link the layers and the dimethyl\u00adformamide solvent mol\u00adecules.In the title compound, {[Cu(C DOI: Click here for additional data file.10.1107/S1600536813006430/hy2619Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The product contains an unusual six-membered thiol\u00adate\u2013carboxyl\u00adate chelate ring. This slightly distorted octa\u00adhedral complex exhibits cis angles ranging from 77.55\u2005(11) to 97.30\u2005(8)\u00b0 due to the presence of the thiol\u00adate\u2013carboxyl\u00adate chelate ring and the constrained bipyridine group. The crystal packing appears to be controlled by a combination of \u03c0-stacking [centroid\u2013centroid distance = 3.611\u2005(2)\u2005\u00c5] and C\u2014H\u22efO inter\u00adactions.The title complex, [Pt(CH DOI: 10.1107/S1600536811013626/tk2735Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, mol\u00adecules are linked by inter\u00admolecular O\u2014H\u22efO hydrogen bonds into chains along [100]. These chains are further linked by weak \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances in the range of 3.609\u2005(2)\u20133.758\u2005(1)\u2005\u00c5, forming a three-dimensional supra\u00admolecular network.In the title centrosymmetric tetra\u00adnuclear complex, [Mn DOI: 10.1107/S1600536810049433/lh5168Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The AgI ion exhibits a slightly distorted tetra\u00adhedral coordination geometry formed by a P atom from a triphenyl\u00adphosphane ligand, two metal-bridging S atoms from thio\u00adsemicabazide ligands and one chloride ion. The S atoms bridge two symmetry-related AgI ions, forming a strictly planar Ag2S2 core with an Ag\u22efAg separation of 2.7802\u2005(7)\u2005\u00c5. There is an intra\u00admolecular N\u2014H\u22efCl hydrogen bond. In the crystal, N\u2014H\u22efCl and N\u2014H\u22efS hydrogen bonds link complex mol\u00adecules, forming layers parallel to (001). These layers are connected through \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.665\u2005(2)\u2005\u00c5], leading to the formation of a three-dimensional network.The dinuclear title complex, [Ag For pot al. 1999; Wujec e al. 2009; Mohareb al. 2009; He et a al. 2012. For exa al. 2012; Lobana al. 2008.2Cl2(CH5N3S)2(C18H15P)2] = 0.026wR(F2) = 0.069S = 1.065026 reflections251 parameters5 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.38 e \u00c5\u22123\u0394\u03c1min = \u22120.55 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536812051562/lh5573Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angle between the central benzene ring and each of the two symmetry-related outer benzene rings is 5.1\u2005(2)\u00b0. The crystal structure is stabilized by inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances in the range 3.466\u2005(2)\u20133.6431\u2005(16)\u2005\u00c5.In the title complex, [Cu(C DOI: 10.1107/S1600536810042789/lh5150Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Within the mononuclear complex mol\u00adecule, intra\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions are observed, the first between a coordinated phen mol\u00adecule and a DHB ligand [centroid\u2013centroid distance = 3.7291\u2005(16)\u2005\u00c5], and the second between a coordinated phen mol\u00adecule and an uncoordinated phen ligand [centroid\u2013centroid distance = 3.933\u2005(2)\u2005\u00c5]. Inter\u00admolecular \u03c0\u2013\u03c0 stacking is observed between adjacent complexes [inter\u00adplanar distance = 3.461\u2005(3)\u2005\u00c5]. Intra- and inter\u00admolecular O\u2014H\u22efO hydrogen bonds are observed in the DHB ligands and between a water mol\u00adecule and DHB ligands, respectively. O\u2014H\u22efN hydrogen bonds are also observed in the DHB ligands and between uncoordinated phen mol\u00adecules and aqua ligands.In the title compound, [La(C DOI: 10.1107/S1600536810047318/vm2058Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Bi\u2014O bond lengths are 2.283\u2005(3) and 2.309\u2005(2)\u2005\u00c5, but as a result of additional long Bi\u22efO inter\u00adactions [2.787\u2005(3) and 2.734\u2005(3)\u2005\u00c5], one of the C\u2014Bi\u2014C angles is 148.62\u2005(13)\u00b0. In the crystal, weak C\u2014H\u22efO hydrogen bonds connect pairs of mol\u00adecules into inversion dimers. These dimers are further connected by weak C\u2014H\u22ef\u03c0 inter\u00adactions into chains along [100] .In the title mol\u00adecule, [Bi(C DOI: Click here for additional data file.10.1107/S1600536813013317/lh5612Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The independent Fe2S2 fragment [Fe\u2014Fe = 2.527\u2005(1)\u2005\u00c5] is in a butterfly conformation, and each Fe atom displays a pseudo-square-pyramidal coordination geometry. The phosphane group occupies an apical position [Fe\u2014P = 2.2670\u2005(14)\u2005\u00c5]. In the crystal, weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into chains along [110]. In the title compound, [Fe DOI: 10.1107/S160053681105584X/cv5221Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CuII ion and the water O atom lie on a twofold axis. The CuII ion exhibits a six-coordinate distorted octa\u00adhedral geometry, with two N atoms from the bipy ligand [Cu\u2014N = 1.9996\u2005(16)\u2005\u00c5] and four O atoms from two 3-meth\u00adoxy\u00adbenzoate ligands [Cu\u2014O = 1.9551\u2005(15) and 2.6016\u2005(16)\u2005\u00c5]. The mol\u00adecules are linked by O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming a three-dimensional network.The title compound, [Cu(C DOI: 10.1107/S1600536811005563/rn2078Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "It has an E conformation with respect to the azomethine double bond and a Z conformation about the amide C=N bond. The CuII atom has a slightly distorted square-pyramidal coordination geometry. The crystal packing involves inter\u00admolecular C\u2014H\u22efO, C\u2014H\u22efN and C\u2014H\u22ef\u03c0 and two types of \u03c0\u2013\u03c0 inter\u00adactions, with centroid\u2013centroid distances of 3.9958\u2005(10) and 3.7016\u2005(13)\u2005\u00c5.The binuclear molecule of the title compound, [Cu DOI: 10.1107/S1600536812031467/fj2577Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The [Co(CN)6]3\u2212 anion exhibits an octa\u00adhedral geometry. In the structure, cations and anions are linked alternatively through O\u2014H\u22efO, O\u2014H\u22efN, N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.523\u2005(2)\u20134.099\u2005(2)\u2005\u00c5] and van der Waals forces, forming a three-dimensional supra\u00admolecular network.The structure of the title compound, (C DOI: Click here for additional data file.10.1107/S1600536813003632/rz5042Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "III compound, {[Sm(C9H7O3)3(H2O)]\u00b7H2O}n, was obtained under hydrothermal conditions. Its structure is isotypic with the analogous Eu complex. The latter was reported incorrectly in space group P1 by Yan et al. . This was corrected by Marsh to P-1. The SmIII ion is nine-coordinated by O atoms from one coordinating water molecule and the remaining ones from the 3-(3-hy\u00addroxy\u00adphen\u00adyl)prop-2-enoatate anions , leading to a distorted tricapped trigonal\u2013prismatic coordination polyhedron surrounded by solvent water mol\u00adecules. In the crystal, extensive intermolecular O\u2014H\u22efO hydrogen-bonding inter\u00adactions and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid separation = 3.9393\u2005(1)\u2005\u00c5] lead to the formation of a three-dimensional supra\u00admolecular network.The title Sm DOI: 10.1107/S1600536812013724/zj2052Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "N. The proposed method has an O(N3 \u00b7 Multiplication(N) + N4) preprocessing time, after which a binomial coefficient C with 0 \u2264 Q \u2264 P \u2264 2N \u2212 1 can be computed modulo 2N in O(N2 \u00b7 log(N) \u00b7 Multiplication(N)) time. Multiplication(N) denotes the time complexity of multiplying two N-bit numbers, which can range from O(N2) to O(N \u00b7 log(N) \u00b7 log(log(N))) or better. Thus, theoverall time complexity for evaluating M binomial coefficients C modulo 2N with 0 \u2264 Q \u2264 P \u2264 2N \u2212 1 is O((N3 + M \u00b7 N2 \u00b7 log(N)) \u00b7 Multiplication(N) + N4). After preprocessing, we can actually compute binomial coefficients modulo any 2R with R \u2264 N. For larger values of P and Q, variationsof Lucas' theorem must be used first in order to reduce the computation tothe evaluation of multiple (O(log\u2061(P))) binomial coefficients C modulo 2N with 0 \u2264 Q\u2032 \u2264 P\u2032 \u2264 2N \u2212 1.I present a new algorithm for computing binomial coefficients modulo 2 N\u2009\u2009(N \u2265 1). The definition of the binomial coefficient C is the usual one:In this paper I present a novel efficient algorithm for computing binomial coefficients modulo 2P \u2264 2N \u2212 1, and after fully handling this case, we will discuss how to compute C modulo 2N for P \u2265 2N.We will mainly consider the case where O(N3 \u00b7 Multiplication(N) + N4) time, where Multiplication(N) is the time complexity for multiplying two N-bit numbers. Multiplication(N) can range from O(N2) to O(N \u00b7 log\u2061\u2061(N) \u00b7 log\u2061\u2061(log\u2061\u2061(N))) or slightly better . FFODD will be equal to the product (modulo\u2009\u20092N) of all the odd numbers from the interval . Thus, we will haveWe will first evaluate FODD is easy to compute. If Q(1) = 0, then FFODD = 1; otherwise it is equal to SOSP(Q(1), 0).FFODD(i > 1. Let X(i) be equal to 2Q(1) + \u22ef+2Q(i\u22121). If Q(i) = 0, then FFODD = (X(i) + 1)\u2009mod\u2061\u20092N. Otherwise we can write FFODD asLet us consider now the case By using Newton's binomial theorem, we can rewrite as(27)\u2211j up to min\u2061{2Q(i)\u22121, N \u2212 1}, because for j \u2265 N, the corresponding term of the sum is zero modulo 2N. We should also notice that SSP(Q(i) \u2212 1, j) is multiplied by 2j. Since its precision is larger than N \u2212 j, the multiplication of SSP(Q(i) \u2212 1, j) and 2j is exact modulo 2N. Then we iterate with j in descending order. For the initial value of j, we compute XP(i) = (X(i)\u22121)j; for the other values, we only multiply XP(i) by (X(i)\u22121)\u22121 in order to obtain XP(i) as (X(i)\u22121)j. We now have an algorithm performing O(N) multiplications of N-bit numbers for computing FFODD. Overall, we have an algorithm performing O(N2) multiplications of N-bit numbers for computing FODD(P) (O(N) multiplications for each of the O(N) bits of P). However, we can do better by rewriting is even. In fact, it is a multiple of 2Q(i\u22121), meaning that it is at least a multiple of 2Q(i)+1. For j > (N/(Q(i) + 1)), each term of the sum will be zero modulo 2N. Thus, we only need to consider at most N/(Q(i) + 1) + 1 terms (from j = 0 to j = N/(Q(i) + 1)). We will start with XP(i) = 1 when j = 0, and then for each subsequent value of j, we will multiply XP(i) by X(i) in order to have XP(i) = X(i)j at each iteration.Note that O(N \u00b7 log\u2061(N)) terms for computing FODD(P)(O(N + N/(Q(2) + 1) + N/(Q(3) + 1)+\u22ef+N/(Q(K) + 1))); for each such term, an N-bit multiplication needs to be performed.Overall we only need to consider P), we can use it in order to write F2(P > 1) asNow that we have a method of efficiently computing FODD(F2(0 \u2264 P \u2264 1) is equal to 1. Since P is of the order of magnitude of 2N and FODD(P) can be evaluated with O(N \u00b7 log\u2061(N))\u2009\u2009N-bit multiplications, computing F2(P) will require O(N2 \u00b7 log\u2061(N))\u2009\u2009N-bit multiplications, obtaining an O(N2 \u00b7 log\u2061(N) \u00b7 Multiplication\u2009(N)) time complexity.C with 0 \u2264 Q \u2264 P \u2264 2N \u2212 1. In order to evaluate it modulo 2N we will first need to compute F2(P), F2(Q), and F2(P \u2212 Q). Then, we will need to find the largest exponent exp\u2061(X) such that 2X)exp\u2061\u2009\u2009N-bit multiplications. Computing exp\u2061(X) where X \u2264 2N \u2212 1 can be performed in O(N2) time (O(N) bit shifts to the right and O(N)\u2009\u2009N-bit number additions). Thus, the time complexity for computing the binomial coefficient modulo 2N is dominated by the computation of F2(P), F2(Q), and F2(P \u2212 Q).As discussed earlier, the multiplicative inverse of an odd number by using , using OC modulo 2N for P \u2265 2N (and 0 \u2264 Q \u2264 P), we need to make use of some variations of Lucas' theorem [P = P1 \u00b7 2N + P0 and Q = Q1 \u00b7 2N + Q0, then, according to [In order to compute a binomial coefficient theorem . If P = rding to , we haveC modulo 2N, we will need to evaluate O(log\u2061(P)) binomial coefficients C, where 0 \u2264 Q\u2032, P\u2032 \u2264 2N \u2212 1. After the O(N3 \u00b7 Multiplication(N)) preprocessing stage, evaluating C modulo 2N will take only O(N2 \u00b7 log\u2061(N) \u00b7 log\u2061(P) \u00b7 Multiplication(N) + log\u20612\u2061(P)) time (the log2(P) factor appears when P \u2265 2N because we need to perform O(log\u2061(P)) divisions, each of which takes O(log\u2061(P)) time because they can be implemented by shifting bits to the right, in order to obtain the O(log\u2061(P)) binomial coefficients or factorials which are needed in order to compute C modulo 2N).Thus, in order to compute C modulo 2N (for P \u2265 2N) to the computation of multiple (O(log\u2061(P))) factorials P\u2032! (of a restricted type) with P\u2032 \u2264 2N \u2212 1 were presented in [Other methods for reducing the computation of ented in .C modulo R, where R is a prime number. The computation of C is reduced to the computation of multiple (O(log\u2061(P))) binomial coefficients C modulo R, with 0 \u2264 Q\u2032 \u2264 P\u2032 \u2264 R \u2212 1. In [M is to evaluate them modulo the prime powers which are divisors of M and then use the Chinese Remained Theorem [M , but inerved in a more derved in .O(log\u20612\u2061(P) + v4 \u00b7 log\u2061(P) \u00b7 log\u2061(u) + v4 \u00b7 u \u00b7 log\u20613\u2061(u)) time for computing C modulo uv, where u is a prime number + N4 \u00b7 log\u2061(P) + N4). If we consider Multiplication(N) = O(N2), our algorithm takes O(N5) time for preprocessing and O(N4 \u00b7 log\u2061(N) \u00b7 log\u2061(P) + log\u20612\u2061(P)) in order to compute C in the general case. These time complexities are slightly worse than the ones obtained in [N) = O(N \u00b7 log\u2061(N) \u00b7 log\u2061(log\u2061(N))) [O(N4 \u00b7 log\u2061(N) \u00b7 log\u2061(log\u2061(N))) time complexity for the preprocessing stage and an O(N3 \u00b7 log\u20612\u2061(N) \u00b7 log\u2061\u2061(log\u2061(N)) \u00b7 log\u2061(P) + log\u20612\u2061(P)) time complexity for actually computing the binomial coefficient modulo 2N. In this case our time complexities are slightly better than the ones presented in [P) > log\u2061(N) \u00b7 log\u2061\u2061(log\u2061(N))). However, it is not clear which time complexity for the multiplication of two N-bit numbers was considered in [An algorithm for computing binomial coefficients modulo prime powers (for any prime) was presented in . The algtated in , but a sained in . Howeverog\u2061(N))) , 2, we oented in .In this paper I presented a novel efficient algorithm for computing binomial coefficients modulo 2The time complexity of the presented algorithm is comparable with that of state-of-the-art algorithms for computing binomial coefficients modulo prime powers . In factN . If the values SSP defined in P, Q) values in a more efficient manner.When computing a small number of binomial coefficients , the bottleneck of the algorithm (in the preprocessing stage) consists of the computation of sums of products of elements of subsets having sizes 0 to"} +{"text": "The dimethyl\u00adamino groups are orientated at 41.80\u2005(7) and 36.43\u2005(7)\u00b0 with respect to the borazine ring. The nitro\u00adoxy group is almost normal to the borazine ring [dihedral angle = 85.33\u2005(14)\u00b0]. The methyl C atom trans to the NO3 group is displaced by \u22120.512\u2005(3)\u2005\u00c5 from the ring plane, whereas the two ortho-methyl C atoms are displaced by 0.239\u2005(3) and 0.178\u2005(3)\u2005\u00c5.In the title compound, C DOI: Click here for additional data file.10.1107/S1600536813007484/hb7055Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536813007484/hb7055Isup3.cmlSupplementary material file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The mean bond lengths in the biferrocene unit are Fe\u2014C = 2.048\u2005(10)\u2005\u00c5 and C\u2014C = 1.427\u2005(8)\u2005\u00c5 within the cyclo\u00adpenta\u00addienyl rings. The B\u2014N bond lengths of the BH3 protected amine is 1.631\u2005(3)\u2005\u00c5. The inter\u00adplanar angle between the two connected cyclo\u00adpenta\u00addienyl rings is 54.29\u2005(8)\u00b0 and the corresponding Fe\u2014Cg\u2014Cg\u2014Fe torsion angle is \u221252.5\u00b0. The conformation of the mol\u00adecule is stabilized by an intra\u00admolecular C\u2014H\u22efBr inter\u00adaction.The title structure, [Fe DOI: 10.1107/S1600536811049270/bq2322Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal packing is controlled by C\u2014H\u22efO hydrogen bonding and \u03c0\u2013\u03c0 stacking inter\u00adactions involving the benzene rings, with a centroid\u2013centroid distance of 3.4220\u2005(1)\u2005\u00c5.In the title compound, (C DOI: 10.1107/S160053681002893X/pv2307Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The amide group is almost perpendicular to the plane of the substituted Cp ring, with a C\u2014N\u2014C\u2014C torsion angle of 101.3\u2005(2)\u00b0, and the N and O atoms in the ethanoyl group are coplanar, with a C\u2014N\u2014C\u2014O torsion angle of \u22120.7\u2005(3)\u00b0. Weak C\u2014H\u22efO hydrogen bonds link adjacent mol\u00adecules.In the title compound, [Fe(C DOI: 10.1107/S1600536812016303/hy2528Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The four O atoms in the equatorial plane around the NiII cation form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 23.67\u2005(8)\u00b0, while the pyridine and benzene rings are oriented at an angle of 89.04\u2005(4)\u00b0. The coordinating water mol\u00adecule links with the carboxyl\u00adate group via an O\u2014H\u22efO hydrogen bond. In the crystal, N\u2014H\u22efO, O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular network. \u03c0\u2013\u03c0 contacts between benzene rings [centroid\u2013centroid distance = 3.8414\u2005(7)\u2005\u00c5] may further stabilize the structure. A weak C\u2014H\u22ef\u03c0 inter\u00adaction also occurs.In the title complex, [Ni(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.023wR(F2) = 0.063S = 1.073492 reflections216 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.40 e \u00c5\u22123\u0394\u03c1min = \u22120.42 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812026943/xu5564Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, there are no hydrogen bonds present, and only very weak \u03c0\u2013\u03c0 inter\u00adactions are observed [centroid\u2013centroid distance = 3.985\u2005(4)\u2005\u00c5], which connect neighbouring tetra\u00adnuclear units into a chain motif along the b axis.In the title centrosymmetric tetra\u00adnuclear complex, [Cu DOI: 10.1107/S1600536811026468/su2284Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atom in the title salt, [Cu(C10H24N4)(H2O)2][CH3(CH2)5CO2]2\u00b72H2O, is chelated by the four N atoms of the 1,4,8,11-tetra\u00adaza\u00adcyclo\u00adtetra\u00addecane (cyclam) ligand and is coordinated by two water mol\u00adecules in a tetra\u00adgonally Jahn\u2013Teller-distorted octa\u00adhedral geometry. The CuII atom lies on a center of inversion. The cations, anions and uncoordinated water mol\u00adecules are linked by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, forming a layer structure parallel to (100). The alkyl chain of the anion is disordered over two positions in a 0.82\u2005(1):0.18\u2005(1) ratio.The Cu DOI: 10.1107/S1600536810025687/bt5285Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In each complex mol\u00adecule, the Cu atom is O,N,O\u2032-chelated by the tridentate Schiff base ligand and N,N\u2032-chelated by the 1,10-phenanthroline ligand in a distorted square-pyramidal geometry. The Cu\u2014N bond distances in the apical directions are 2.298\u2005(4) and 2.268\u2005(4)\u2005\u00c5. In the crystal, inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds together with C\u2014H\u22ef\u03c0 inter\u00adactions result in a three-dimensional supra\u00admolecular structure.The asymmetric unit of the title complex, [Cu(C DOI: 10.1107/S160053681004554X/xu5077Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII atom is located about an inversion centre. The mol\u00adecule consists of three planar units , adopting a staircase-like structure. The six-membered metallocycles have a sofa conformation with the Cu atom out of the plane of the 1,3,5-triaza\u00adpenta\u00addienyl ligands by 0.246\u2005(5)\u2005\u00c5. The ipso-C atoms of the CCl3 substituents are slightly out of the 1,3,5-triaza\u00adpenta\u00addienyl planes by 0.149\u2005(6) and \u22120.106\u2005(6)\u2005\u00c5. The CCl3 groups of each 1,3,5-triaza\u00adpenta\u00addienyl ligand are practically in the energetic\u00adally favourable mutually eclipsed conformation. In the crystal, the mol\u00adecules are packed in stacks along the a axis. The mol\u00adecules in the stacks are held together by two additional axial Cu\u22efCl inter\u00adactions of 3.354\u2005(2)\u2005\u00c5. Taking the axial Cu\u22efCl inter\u00adactions into account, the CuII atom exhibits a distorted [4\u00a0+\u00a02]-octa\u00adhedral coordination environment. The stacks are bound to each other by weak inter\u00admolecular attractive Cl\u22efCl [3.505\u2005(2)\u20133.592\u2005(3)\u2005\u00c5] inter\u00adactions.The title compound, [Cu(C DOI: 10.1107/S1600536812036124/aa2069Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The mol\u00adecule is located about a twofold rotation axis. The coordination environment of the SnIV atom is a distorted octa\u00adhedron, with two bidentate 2-pyridine\u00adseleno\u00adlate ligands inclined to each other at an angle of 83.96\u2005(7)\u00b0. The two Sn\u2014Cl and two Sn\u2014N bonds are in cis configurations, while the two Sn\u2014Se bonds of 2.5917\u2005(3)\u2005\u00c5 are in a trans configuration, with an Se\u2014Sn\u2014Se angle of 157.988\u2005(15)\u00b0. In the crystal, \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings [centroid-to-centroid distance of 3.758\u2005(3)\u2005\u00c5] and weak inter\u00admolecular C\u2014H\u22efCl hydrogen bonds link the mol\u00adecules into chains along the c axis.The title compound, [SnCl DOI: 10.1107/S1600536813014657/cv5413Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Each AgI ion is coordinated by two N atoms from two bridging bipy ligands, forming chains along [101]. \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings [centroid\u2013centroid distances = 3.638\u2005(8) and 3.688\u2005(8)\u2005\u00c5] connect the chains. Inter\u00admolecular O\u2014H\u22efO hydrogen bonds link the uncoord\u00adinated water mol\u00adecules and the perchlorate anions.In the title compound, {[Ag DOI: 10.1107/S1600536811040153/hy2472Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One of the two I\u2212 anions is disordered over two sets of sites, with refined occupancies of 0.66\u2005(5) and 0.34\u2005(5). The CdII ions are bridged by bmi ligands, leading to the formation of a chain along [001]. Dimethyl\u00adformamide solvent mol\u00adecules are located between these chains. Classical N\u2014H\u22efO hydrogen bonding between the bmi ligands and the solvent mol\u00adecules leads to a consolidation of the structure.In the title complex, {[CdI N-heterocyclic ligands, see: Meng et al. ]\u00b7C3H7NO = 0.035 wR(F 2) = 0.071 S = 1.15 3950 reflections218 parametersH-atom parameters constrainedmax = 0.42 e \u00c5\u22123 \u0394\u03c1min = \u22120.77 e \u00c5\u22123 \u0394\u03c1 CrystalClear used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811050823/wm2569Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The bond lengths and angles (C\u2014Fe\u2014Fe and Fe\u2014S\u2014Fe) are within expected ranges. The S\u22efS distance [2.9069\u2005(12)\u2005\u00c5] and the dihedral angle between two Fe\u2014S\u2014Fe planes [78.5\u2005(3)\u00b0] of the butterfly-shaped Fe2S2 core are enlarged compared with related bridged dithiol\u00adate diiron analogues. The calculated 4-benzothia\u00adzolebenzyl best planes are almost parallel [dihedral angle = 3.7\u2005(7)\u00b0].The title compound, [Fe DOI: 10.1107/S1600536812007581/vm2158Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "A short Cd\u22efN contact [2.890\u2005(3)\u2005\u00c5] to the monodentate sulfadiazinate ion also occurs. The dihedral angles between the planes of the aromatic rings of the anions are 86.81\u2005(14) and 68.65\u2005(14)\u00b0. The crystal structure features inter\u00admolecular N\u2014H\u22efO, O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds.In the title compound, [Cd(C DOI: 10.1107/S1600536811019635/hb5888Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The employed \u03b1-diimine opens up, forming a highly asymmetric ammonium that has its protons at one of the N atoms [N\u2014C= 1.264\u2005(4) and 1.516\u2005(4)\u2005\u00c5]. One of the C=N double bonds was oxidized to C\u2014N, which is consistent with the bond length of 1.516\u2005(4)\u2005\u00c5. Meanwhile SnIV was reduced to SnII. The (SnCl)3\u2212 anion is trigonal\u2013pyramidal. In the crystal, mol\u00adecules are linked by C\u2014H\u22efCl, N\u2014H\u22efCl, N\u2014H\u22efN and C\u2014H\u22efN bonds. The crystal studied was twinned by pseudo-merohedry.In the title compound, (C DOI: 10.1107/S1600536812014729/hp2032Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The NiII ion is six-coordinated in a distorted octa\u00adhedral geometry by two N, two O and two S atoms. An intra\u00admolecular C\u2014H\u22efS hydrogen bond generates an S(6) ring motif. In the crystal, mol\u00adecules are linked through inter\u00admolecular N\u2014H\u22efS, N\u2014H\u22efO, C\u2014H\u22efN, C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds into infinite two-dimensional network parallel to the ab plane. The structure is further stablized by weak C\u2014H\u22ef\u03c0 inter\u00adactions. The dimethylformamide solvent molecule is disordered over two sets of sites in a 0.514\u2005(15):0.486\u2005(15) ratio.The asymmetric unit of the title compound, [Ni(C DOI: 10.1107/S1600536812012834/zj2062Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Two O atoms of one bidentate 2-meth\u00adoxy\u00adbenzoate ligand are each disordered over two positions, with site-occupancy factors of 0.579\u2005(4) and 0.421\u2005(4). In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, forming a two-dimensional network lieing parallel to the bc plane. The crystal packing is further stablized by \u03c0\u2013\u03c0 stacking inter\u00adactions between the dmphen rings of neighboring mol\u00adecules, with distances between their parallel dmphen ring planes of 3.517\u2005(3) and 3.610\u2005(3)\u2005\u00c5.In the title compound, [Cd(C DOI: 10.1107/S1600536812010835/bg2438Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The asymmetric unit consists of half a Lindquist anion and one [Cu(phen)(PPh3)2]+ cationic complex. In the cation, there are intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.617\u2005(2) and 3.7272\u2005(18)\u2005\u00c5]. This inorganic\u2013organic adduct is connected by C\u2014H\u22efO hydrogen bonds, forming a two dimensional network lying in the ab plane. These networks are connected by C\u2014H\u22ef\u03c0 inter\u00adactions into a three-dimensional structure.The title compound, [Cu(C DOI: 10.1107/S1600536812036367/hg5234Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Extensive inter\u00admolecular N\u2014H\u22efO, N\u2014H\u22efN, O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds, as well as weak inter\u00admolecular C\u2014H\u22efN and C\u2014H\u22efO inter\u00adactions occur in the crystal. \u03c0\u2013\u03c0 stacking between between pyrazine rings of coordinating ligands and lattice molecules [centroid\u2013centroid distance = 3.5669\u2005(14)\u2005\u00c5] may further stabilize the structure.In the title compound, [Cd(NO DOI: 10.1107/S1600536812028577/xu5577Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Fe\u2014O bond lengths are in the range 1.9818\u2005(18)\u20131.9957\u2005(18)\u2005\u00c5. The trans O\u2014Fe\u2014O angles are 169.06\u2005(13) and 171.54\u2005(8)\u00b0, whereas the corresponding cis angles are in the range 84.81\u2005(10)\u2013100.68\u2005(12)\u00b0. In the crystal, mol\u00adecules are linked via C\u2014H\u22efCl inter\u00adactions.In the title compound, [Fe(C DOI: 10.1107/S1600536812023215/im2375Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Cu\u2014N bond lengths in the base of the resulting CuN5 pyramid are in the range 2.016\u2005(3)\u20132.024\u2005(3)\u2005\u00c5 and the apical Cu\u2014N(\u00a0C) distance is 2.385\u2005(4)\u2005\u00c5. Based on the \u03c4 parameter, the shape of the coordination polyhedron is tetra\u00adgonal\u2013pyramidal (\u03c4 = 0). All atoms of the square-planar tetracyanidopalladate(II) moiety and the CuII ion are located on a mirror plane. The electroneutral mol\u00adecules inter\u00adact by N\u2014H\u22efN hydrogen bonds, resulting in the formation of a three-dimensional network.The title compound, [Cu(NH II complexes see: Escorihuela et al. 4] = 0.025wR(F2) = 0.061S = 1.001051 reflections81 parametersH-atom parameters constrainedmax = 0.54 e \u00c5\u22123\u0394\u03c1min = \u22121.26 e \u00c5\u22123\u0394\u03c1X-AREA used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536813011033/ff2104Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The anions are disordered each over two sets of sites, with an occupancy ratio of 0.617\u2005(6):0.383\u2005(6). The distances from the Fe atoms to the centroids of the unsubstituted and substituted cyclo\u00adpenta\u00addienyl (Cp) rings are 1.645\u2005(1)/1.657\u2005(1) and 1.644\u2005(1)/1.647\u2005(1)\u2005\u00c5. The dihedral angles between the two Cp rings are 2.49\u2005(3) and 1.45\u2005(4)\u00b0 in the two ferrocenyl groups of the cations.The asymmetric unit of the title complex, [Fe(C DOI: 10.1107/S1600536812001766/hy2506Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In each mol\u00adecule, four penta\u00adfluoro\u00adbenzoate anions bridge the quadruply bonded Mo2 4+ unit that is, in addition, axially coordinated by two O atoms of tetra\u00adhydro\u00adfuran (THF) mol\u00adecules. In the two independent mol\u00adecules, the mean Mo\u2014Mo bond length is 2.110\u2005\u00c5. Since the THF mol\u00adecules are equally disordered over two sets of sites, there are four different Mo\u2014O distances in both half-mol\u00adecules with an overall mean of 2.542\u2005\u00c5. A zigzag chain is formed by \u03c0\u2013\u03c0 stacking inter\u00adactions between penta\u00adfluoro\u00adphenyl rings, indicated by a centroid\u2013centroid distance of 3.7054\u2005(11)\u2005\u00c5 and a centroid-to-plane distance of 3.4169\u2005(3)\u2005\u00c5. The extension of the unit gives a three-dimensional network structure with the THF mol\u00adecules located in the voids.In the asymmetric unit of the title compound, [Mo DOI: 10.1107/S1600536811033411/wm2516Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The four O atoms in the equatorial plane around the CoII cation form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 22.3\u2005(3)\u00b0, while the pyridine ring and the benzene ring are oriented at a dihedral angle of 84.59\u2005(13)\u00b0. Intra\u00admolecular O\u2014H\u22efO hydrogen bonding occurs between the carboxyl\u00adate group and coordinated water mol\u00adecule. In the crystal, N\u2014H\u22efO, O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular network.In the title complex, [Co(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.046wR(F2) = 0.110S = 1.163618 reflections203 parameters6 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 2.60 e \u00c5\u22123\u0394\u03c1min = \u22122.03 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S160053681201330X/xu5493Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The FeIII cation has an approximate octa\u00adhedral geometry, defined by six N atoms from two bpca\u2212 anions. The nearly parallel [dihedral angle = 1.50\u2005(1)\u00b0] bpca\u2212 anions form two-dimensional supermolecules along the a axis by the way of weak \u03c0\u2013\u03c0 stacking inteactirons . Intra- and inter\u00admolecular C\u2014H\u22efO hydrogen bonding occurs. The water mol\u00adecule [occupancies 0.520\u2005(5) and 0.480\u2005(5)], two carbonyl O atoms [occupancies 0.622\u2005(7) and 0.378\u2005(7)] and the four perchlorate O atoms [occupancies 0.887\u2005(4) and 0.113\u2005(4)] are each disordered over two positions.The structure of the title salt complex, [Fe(C DOI: 10.1107/S1600536811047684/jj2105Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, inter\u00admolecular O\u2014H\u22efO hydrogen bonds lead to a three-dimensional supra\u00admolecular network. Intra\u00admolecular O\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings and between the benzene and triazole rings [centroid\u2013centroid distances = 3.657\u2005(1) and 3.752\u2005(1)\u2005\u00c5] are observed.In the title compound, [Cu(C DOI: 10.1107/S1600536811018356/hy2429Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The average Ru\u2014S bond length is 2.41\u2005(1)\u2005\u00c5 and the average S\u2014Ru\u2014S bite angle is 81.13\u2005(19)\u00b0.In the title complex, [Ru(C DOI: Click here for additional data file.10.1107/S1600536813014141/hy2626Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The packing of the structure involves weak \u03c0\u2013\u03c0 inter\u00adactions between the pyridyl and benzene rings of neighbouring ppaBr ligands [centroid\u2013centroid distance = 3.928\u2005(2)\u2005\u00c5] and inter\u00adactions between the Br atom on the ppaBr ligand and the hfac ligand [Br\u22efC = 3.531\u2005(2)\u2005\u00c5].In the title complex, [Co(C DOI: 10.1107/S1600536810032757/zs2056Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Co2+ ion, coordinated by three O atoms from two different carboxyl\u00adate groups and three water mol\u00adecules, displays a distorted octa\u00adhedral environment. In the crystal, \u03c0\u2013\u03c0 inter\u00adchain inter\u00adactions, with centroid\u2013centroid distances of 3.656\u2005(2) and 3.669\u2005(2)\u2005\u00c5 between the benzene rings of the ligands, assemble the chains into sheets parallel to (100). O\u2014H\u22efO hydrogen-bonding inter\u00adactions between the coordinating water mol\u00adecules and carboxyl\u00adate O atoms link the sheets into a three-dimensional network.The polymeric title compound, {[Co(C DOI: 10.1107/S1600536812009580/wm2597Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The crystal structure is made up of \u221e 1[Nb2PS10] chains expanding along [010]. These chains are built up from bicapped trigonal-prismatic [Nb2S12] units and tetra\u00adhedral [PS4] groups and are linked through a linear S\u2014Ag\u2014S bridge, forming a two-dimensional layer. These layers then stack on top of each other, completing the three-dimensional structure with an undulating van der Waals gap. The disordered Cs+ ions reside on sites with half-occupation in the voids of this arrangement. Short [2.8843\u2005(5)\u2005\u00c5] and long [3.7316\u2005(4)\u2005\u00c5] Nb\u2014Nb distances alternate along the chains, and anionic S2 2\u2212 and S2\u2212 species are observed. The charge balance of the com\u00adpound can be represented by the formula [Cs+]0.5[Ag+]0.5[Nb4+]2[PS4 3\u2212][S2 2\u2212]3.The quinter\u00adnary thio\u00adphosphate Cs DOI: 10.1107/S1600536810021768/wm2357Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the coordination cation, the CuII atom is coordinated by four N atoms from two 1,10-phenanthroline ligands and two O atoms from a bridging\u2013chelating carboxyl\u00adate group of the pyridine-2,6-dicarboxyl\u00adate anion, completing a distorted N4O2 octa\u00adhedral coordination environment. The TbIII atom, located on a twofold rotation axis, is nine-coordinated by three tridentate pyridine-2,6-dicarboxyl\u00adate anions forming an N3O6 donor set. The intra\u00admolecular Cu\u22efTb distance of 5.0592\u2005(11)\u2005\u00c5 indicates weak inter\u00adactions between the CuII and TbIII atoms. The coordination cations, nitrate anions and water mol\u00adecules are connected via O\u2014H\u22efO hydrogen bonds into layers parallel to the (001) plane. Moreover, there are extensive \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 4.332\u2005(7) and 3.878\u2005(5)\u2005\u00c5] between the phenanthroline ligands and between phenanthroline and pyridine-2,6-dicarboxyl\u00adate ligands.The asymmetric unit of the title compound, [Cu For the al. 2010; Yang et al. 2006.2Tb(C7H3NO4)3(C12H8N2)4]NO3\u00b74H2O = 0.041wR(F2) = 0.134S = 1.046454 reflections479 parameters27 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.88 e \u00c5\u22123\u0394\u03c1min = \u22121.09 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812031686/gk2483Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The central ion is coordinated by the thia\u00adzole N atom, the thio\u00adureido N and the S atom of the deprotonated thio\u00adsemicarbazone ligand. The other ligand is linked with the central ion through the C=S group. The deprotonated ligand intra\u00admolecularly hydrogen bonds to the thia\u00adzole ring N atom, while the ligand forms an inter\u00admolecular hydrogen bond to the thiol\u00adate S atom of the second ligand. The deprotonation of the tridentate ligand and its coordination to the CdII ion via the S atom strikingly affects the C\u2014S bond lengths. The C\u2014S bond lengths in the neutral and deprotonated ligands in the metal complex are 1.709\u2005(3) and 1.748\u2005(2)\u2005\u00c5, respectively, whereas it is 1.671\u2005(3)\u2005\u00c5 in the free ligand. In the metal complex, the Cd\u2014S distances are 2.6449\u2005(6) and 2.5510\u2005(6)\u2005\u00c5. The Cd\u2014I bond length is 2.7860\u2005(2)\u2005\u00c5.In the title complex, [Cd(C I(C12H12N4S2)] = 0.027wR(F2) = 0.060S = 1.028142 reflections354 parameters3 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.92 e \u00c5\u22123\u0394\u03c1min = \u22120.60 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 global, I. DOI: Click here for additional data file.10.1107/S160053681300915X/bv2219Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The distorted trans square-planar environment is completed by two P atoms [Rh\u2014P = 2.3251\u2005(4)\u2005\u00c5] from two diphen\u00adyl(4-vinyl\u00adphen\u00adyl)phosphane ligands. The vinyl group is disordered over two sets of sites in a 0.668\u2005(10):0.332\u2005(10) ratio. The crystal packing exhibits weak C\u2014H\u22efCl and C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the phenyl rings of neighbouring mol\u00adecules, with a centroid\u2013centroid distance of 3.682\u2005(2)\u2005\u00c5.In the title compound, DOI: 10.1107/S1600536812013669/cv5270Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CdII ion is in a strongly distorted CdN2O5 penta\u00adgonal-bipyramidal geometry, defined by two N atoms from one 2,2\u2032-bipyridine ligand and five O atoms from three 2-phenyl\u00adquinoline-4-carboxyl\u00adate ligands, one monodentate, two bidentate. Weak inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.712\u2005(3)\u2005\u00c5] help to establish the packing of the structure.The neutral binuclear title complex, [Cd DOI: 10.1107/S1600536810049640/bh2323Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the components are linked into a three dimensional network by O\u2014H\u22efO, N\u2014H\u22efO, N\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and a \u03c0\u2013\u03c0 inter\u00adaction with a centroid\u2013centroid distance of 3.6080\u2005(8)\u2005\u00c5.In the complex anion of the title compound, (C DOI: 10.1107/S1600536811011147/is2693Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, noncovalent inter\u00adactions play an important role in the stabilization of the structure, involving O\u2014H\u22efO, N\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings of the pyridine-2,6-dicarboxyl\u00adate ligands [centroid\u2013centroid distance = 3.7138\u2005(15)\u2005\u00c5] and between the 8-hy\u00addroxy-2-methyl\u00adquinolinium cations .In the title compound, (C DOI: 10.1107/S1600536811021015/hy2430Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Ni\u2014N distances range from 2.0594\u2005(12) to 2.0664\u2005(12)\u2005\u00c5, intra-ligand N\u2014Ni\u2014N angles range from 84.59\u2005(5) to 86.06\u2005(5)\u00b0, and adjacent inter-ligand N\u2014Ni\u2014N angles range between 93.94\u2005(5) and 95.41\u2005(5)\u00b0. In the crystal, inversion-related pyrazole rings are \u03c0\u2013\u03c0 stacked, with an inter\u00adplanar spacing of 3.4494\u2005(18)\u2005\u00c5, forming chains that propagate parallel to the a-axis direction. Inter\u00admolecular O\u2014H\u22efO hydrogen bonds are present between water mol\u00adecules and tri\u00adfluoro\u00admethane\u00adsulfonate anions.In the title salt, [Ni(C DOI: 10.1107/S1600536813024252/tk5252Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CuII atom is in a square-pyramidal coordination environment formed by four O atoms of four carboxyl\u00adate groups and one water mol\u00adecule, which is located in the apical position. The carboxyl\u00adate groups are twisted relative to the benzene rings by 11.09\u2005(16) and 45.55\u2005(19)\u00b0. The nitro groups are not coplanar with the parent aromatic rings [dihedral angles = 16.2\u2005(3)\u201351.45\u2005(14)\u00b0]. O\u2014H\u22efO hydrogen bonds between the coordinated water mol\u00adecules and one of the nitro groups, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.5764\u2005(12)\u2005\u00c5] between the benzene rings, assemble the complex mol\u00adecules into a one-dimensional polymeric structure which is further extended into a three-dimensional polymeric network via O\u2014H\u22efO hydrogen bonds involving the water molecules of crystallization.The title compound, [Cu DOI: 10.1107/S1600536811000547/gk2339Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The first reported polymorph of this complex was triclinic . The present polymorph crystallized in the monoclinic space group P21/c. The CuII ion is in a square planar environment and is coordinated by one phenolate O, one imine N and one pyridine N atom of the tridentate Schiff base ligand and by one water O atom. In the crystal, mol\u00adecules are linked through inter\u00admolecular O\u2014H\u22efO hydrogen bonds to form chains along the a axis.The title complex, [Cu(C al. 2010. Acta Cr For the al. 2010.13H10ClN2O)(H2O)]NO3\u00b7H2O = 0.037wR(F2) = 0.089S = 1.063410 reflections233 parameters3 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.39 e \u00c5\u22123\u0394\u03c1min = \u22120.48 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXTL I, global. DOI: 10.1107/S1600536812004564/qm2051Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The B atom deviates by 0.202\u2005(1)\u2005\u00c5 from the least-squares plane of the mesityl ring. All of the meth\u00adoxy groups are approximately coplanar with the 2,4,6-trimeth\u00adoxy\u00adphenyl ring, whereas the BOH group is twisted with respect to it by 19.5\u00b0. The borinic OH group is engaged in an intra\u00admolecular O\u2014H\u22efO hydrogen bond with one of ortho-meth\u00adoxy groups. The mol\u00adecular structure is stabilized by weak C\u2014H\u22efO contacts. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions, generating a three-dimensional network.In the title mol\u00adecule, C DOI: 10.1107/S160053681002297X/pv2296Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One CdII ion is six-coordinated and exhibits a distorted octa\u00adhedral geometry, while the other two CdII ions are seven-coordinated, displaying a distorted penta\u00adgonal\u2013bipyramidal geometry. The CdII ions are bridged by two types of sulfate anions, producing inorganic chains along [100]. These chains are further connected by the H2bic ligands, leading to a three-dimensional framework. N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the imidazole and benzene rings further stabilize the crystal structure. The asymmetric unit of the title compound, [Cd H-benzimidazole-5-carboxyl\u00adate com\u00adplexes, see: Gao et al. 2(SO4)2(H2O)3] = 0.025 wR(F 2) = 0.067 S = 1.06 3935 reflections361 parameters1 restraintH-atom parameters constrainedmax = 0.56 e \u00c5\u22123 \u0394\u03c1min = \u22120.71 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811034477/hy2460Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Ir=Ccarbene bond is strong and short and exerts a notable effect on the trans-Ir\u2014N bond, which is about 0.10\u2005\u00c5 longer than the two other Ir\u2014N bonds. The chloro\u00adform solvent mol\u00adecule is anchored via a weak C\u2014H\u22efCl hydrogen bond to the Cl atom of the Ir complex mol\u00adecule. In the crystal, the constituents adopt a layer-like arrangement parallel to (010) and are held together by weak inter\u00admolecular C\u2014H\u22efCl hydrogen bonds, as well as weak Cl\u22efCl [3.498\u2005(2)\u2005\u00c5] and Cl\u22ef\u03c0 [3.360\u2005(4)\u2005\u00c5] inter\u00adactions. A weak intra\u00admolecular C\u2014H\u22efO hydrogen bond is also observed.In the title compound, [Ir(C N,N\u2032,N\u2032\u2032)-iridium moiety Ir[TpMe2]. Its formation from [(TpMe2)Ir(C6H5)2(k1-N2)] and eth\u00adoxy\u00adbenzene involved multiple C\u2014C,H,O,Cl bond transformations by the outstanding activity of the Ir[TpMe2] moiety. For general information on C\u2014H and C\u2014C activation, see: Lin & Yamamoto borate-moto 1999; Dyker (C8H7O)Cl]\u00b7CHCl3 = 0.028wR(F2) = 0.071S = 1.028053 reflections341 parametersH-atom parameters constrainedmax = 1.29 e \u00c5\u22123\u0394\u03c1min = \u22121.43 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536813007344/lh5594Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Both the arsine and phosphine ligands are equatorial with respect to the Ru3 triangle. In addition, each Ru atom carries one equatorial and two axial terminal carbonyl ligands. In each biphenyl unit, the phenyl rings are twisted from each other, making dihedral angles of 51.22\u2005(18), 42.94\u2005(16) and 26.95\u2005(16)\u00b0. The arsine-substituted phenyl rings make dihedral angles of 61.22\u2005(15), 87.17\u2005(15) and 83.32\u2005(15)\u00b0 with each other. The dihedral angles between the two benzene rings are 85.52\u2005(18) and 81.77\u2005(15)\u00b0 for the two diphenyl\u00adphosphanyl groups, respectively. In the crystal, mol\u00adecules are linked into dimers by inter\u00admolecular C\u2014H\u22efO hydrogen bonds. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 [centroid\u2013centroid distance = 3.6981\u2005(18)\u2005\u00c5] inter\u00adactions stabilize the crystal structure.In the title DOI: 10.1107/S160053681100078X/ng5095Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The structure of the dinuclear tetracation exhibits significant distortion from planarity in the bridging 2,3,5,6-tetra\u00adkis\u00ad(pyridin-2-yl)pyrazine (tppz) ligand, which has a saddle-like geometry with an average dihedral angle of 42.96\u2005(18)\u00b0 between adjacent pyridine rings. The metal\u2013ligand coordination environment is nearly equivalent for the two RuII atoms, which have a distorted octa\u00adhedral geometry due to the restricted bite angle [157.57\u2005(13)\u2013159.28\u2005(12)\u00b0] of their two mer-arranged tridendate ligands orthogonal to each other. At the peripheral tpy ligands, the average Ru\u2014N bond distance is 2.072\u2005(4)\u2005\u00c5 for the outer N atoms trans to each other (Nouter) and 1.984\u2005(1)\u2005\u00c5 for the central N atoms . At the bridging tppz ligand, the average metal\u2013ligand distances are significantly shorter [2.058\u2005(4)\u2005\u00c5 for Ru\u2014Nouter and 1.965\u2005(1)\u2005\u00c5 for Ru\u2014Ncentral] as a result of both the geometric constraints and the stronger \u03c0-acceptor ability of the pyrazine-centered bridge. The dihedral angle between the two tpy planes is 27.11\u2005(6)\u00b0. The intra\u00admolecular linear distance between the two Ru atoms is 6.6102\u2005(7)\u2005\u00c5.In the title compound [Ru DOI: Click here for additional data file.10.1107/S1600536812051215/zl2523Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The binuclear complex mol\u00adecules are linked together by inter\u00admolecular O\u2014H\u22efO hydrogen bonds into a layer parallel to (100). The layers are connected by C\u2014H\u22efCl hydrogen bonds. Intra\u00admolecular O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.5958\u2005(16)\u2005\u00c5] are also present.In the centrosymmetric binuclear title compound, [Cu DOI: 10.1107/S1600536811035112/hy2462Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, weak N\u2014H\u22efCl hydrogen bonds link the mol\u00adecules into layers, while weak \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 4.2758\u2005(18)\u2005\u00c5] also help to stabilize the packing.In the title compound, (C DOI: 10.1107/S1600536810046817/jh2229Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adaction [centroid\u2013centroid distance = 3.6113\u2005(11)\u2005\u00c5] between two of the 4-fluoro\u00adphenyl groups is complemented by both C\u2014H\u22efF and C\u2014H\u22efO inter\u00adactions with distances in the range 2.51\u20132.60\u2005\u00c5, resulting in a tight head-to-tail packing. In the title complex, [Cu(NO DOI: Click here for additional data file.10.1107/S1600536812043346/mw2090Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The sixth coordination site of molybdenum is occupied by the O atom of a methanol mol\u00adecule. In this complex, the NO5 coordination sphere adopts a distorted octa\u00adhedral coordination geometry. The metal atom is shifted by 0.335\u2005(1)\u2005\u00c5 from the square plane defined by the three donor atoms of the Schiff base ligand and one oxide group towards the second oxide group in the cis position. In the crystal, the complex forms inversion dimers through a pair of O\u2014H\u22efN hydrogen bonds involving the methanol \u2013OH group and the pyridine N atom. Additional C\u2014H\u22efO contacts stack the mol\u00adecules along the b axis.In the title complex, [Mo(C O2(CH4O)] = 0.026wR(F2) = 0.070S = 1.073474 reflections239 parameters13 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.31 e \u00c5\u22123\u0394\u03c1min = \u22120.60 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXTL I, global. DOI: 10.1107/S1600536813019077/sj5344Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The bridging carbonyl C\u2014C(=O)\u2014C plane makes dihedral angles of 47.07\u2005(6)\u00b0 with the naphthalene ring system and 24.20\u2005(10)\u00b0 with the benzene ring. A weak inter\u00admolecular C\u2014H\u22efO hydrogen bond exists between the H atom of one meth\u00adoxy group and the O atom of the other meth\u00adoxy group in an adjacent mol\u00adecule. The crystal packing is additionally stabilized by two types of weak inter\u00admolecular inter\u00adactions involving the Br atom, C\u2014H\u22efBr and Br\u22efO [3.2802\u2005(14)\u2005\u00c5].In the title compound, C DOI: 10.1107/S160053681004016X/vm2049Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The central PbII ion has a (4\u00a0+\u00a02) coordination by four O atoms of the two ABTC ligands and two weaker Pb\u2014S bonding inter\u00adactions (Pb\u2014S secondary bonds) from S atoms of other two neighbouring ABTC ligands. These bonds link the metal ions into zigzag chains along the c axis, which, in turn, aggregate through \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.7436\u2005\u00c5] between ABTC rings and N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds.The title complex, [Pb(C DOI: 10.1107/S1600536810049330/bg2378Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There are C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.319\u2005(1)\u2005\u00c5] in the crystal structure.In the title compound, [Cd(NO DOI: 10.1107/S1600536811039687/jh2323Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The anion is a six-coordinated complex with a distorted CuN2O4 octa\u00adhedral geometry around the CuII ion. N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds along with \u03c0\u2013\u03c0 contacts between the pyridine rings of the (2a4mpH)+ cations [centroid\u2013centroid distance = 3.573\u2005(2)\u2005\u00c5] stabilize the crystal structure.The asymmetric unit of the title compound, (C DOI: 10.1107/S1600536811001139/bt5449Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecular complex has no crystallographically imposed symmetry. The coordination geometry around the metal atom is distorted square-pyramidal. The equatorial coordination around copper involves donor atoms of the bidentate chelating 2-(dimethyl\u00adamino)\u00adethanol ligand and the 2-(dimethyl\u00adamino)\u00adethano\u00adlate group, which are mutually trans to each other, with four approximately equal short Cu\u2014O/N bond distances. The axial Cu\u2014I bond is substanti\u00adally elongated. Inter\u00admolecular hydrogen-bonding inter\u00adactions involving the \u2013OH group of the neutral 2-(dimethyl\u00adamino)\u00adethanol ligand to the O atom of the monodeprotonated 2-(dimethyl\u00adamino)\u00adethano\u00adlate group of the mol\u00adecule related by the n-glide plane, as indicated by the O\u22efO distance of 2.482\u2005(12)\u2005\u00c5, form chains of mol\u00adecules propagating along [101].The title compound, [Cu(C I(C4H11NO)] = 0.058wR(F2) = 0.204S = 1.002471 reflections133 parametersH-atom parameters constrainedmax = 1.80 e \u00c5\u22123\u0394\u03c1min = \u22120.83 e \u00c5\u22123\u0394\u03c1AFC6S Diffractometer Control Software AFC6S Diffractometer Control Software; data reduction: TEXSAN I, global. DOI: 10.1107/S1600536812010215/ds2178Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "An intra\u00admolecular O\u2014H\u22efO hydrogen bond in the ligand generates an S(6) ring. The dihedral angle between the aromatic rings joined to the acetyl\u00adacetonate unit is 6.4\u2005(2)\u00b0. The ethanol mol\u00adecule is disordered over two orientations in a 0.65\u2005(3):0.35\u2005(3) ratio. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO bonds.In the title complex, [Co(C DOI: 10.1107/S1600536810029776/hb5579Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the ferrocenyl moiety, the unsubstituted cyclo\u00adpenta\u00addienyl ring is disordered over two orientations with site occupancies of 0.64\u2005(2) and 0.36\u2005(2). In the pyrrolizine ring, one C atom is disordered over two positions, with site occupancies of 0.71\u2005(1) and 0.29\u2005(1). Intra\u00admolecular C\u2014H\u22efO inter\u00adactions occur. The crystal packing is established through weak inter\u00admolecular C\u2014H\u22efO and N\u2014H\u22efO inter\u00adactions.In the title compound, [Fe(C DOI: 10.1107/S1600536810045459/im2240Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The four acetamide ligands bridging the dirhodium core are arranged in a 2,2-cis manner, with two N atoms and two O atoms coordinating to the unique RhII atom cis to one another. The Neq\u2014Rh\u2014Rh\u2014Oeq torsion angles on the acetamide bridges vary between 1.62\u2005(4) and 1.78\u2005(4)\u00b0. The Rh\u2014Rh bond length is 2.4319\u2005(3)\u2005\u00c5. The axial nitrile ligand completes the distorted octahedral coordination sphere and shows a non-linear coordination with an Rh\u2014N\u2014C bond angle of 167.14\u2005(15)\u00b0, while the N\u2014C bond length is 1.135\u2005(3)\u2005\u00c5.The complex molecule of the title compound, [Rh DOI: Click here for additional data file.10.1107/S1600536813012828/mw2103Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The AuI atom is in a trigonal\u2013planar geometry within an IP2 donor set with the greatest distortion seen in the P\u2014Au\u2014P angle [128.49\u2005(3) \u00b0]. Close intra\u00admolecular Au\u22efO inter\u00adactions [3.172\u2005(3)\u2005\u00c5] are observed. No specific inter\u00admolecular inter\u00adactions are noted in the crystal packing.In the title compound, [AuI(C DOI: 10.1107/S1600536812011609/su2394Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The average of the four Ru\u2014Cl bond lengths is 2.355\u2005(15)\u2005\u00c5, and the Ru\u2014S and Ru\u2014N bond lengths are 2.2853\u2005(3) and 2.1165\u2005(11)\u2005\u00c5, respectively. The complex forms a chain, with a six-coordinate sodium ion bridging the ruthenium(III) units. The sodium cation is coordinated by cis-chloride ligands on ruthenium [Na\u2014Cl = 2.9576\u2005(7) and 2.6988\u2005(7)\u2005\u00c5], chloride and DMSO ligands from the ruthenium complexes related by inversion , a nitro\u00adgen ligand from the pyrimidine of the tetrachlorido\u00adruthenium(III) complex related by the twofold rotation axis [Na\u2014N = 2.5224\u2005(14)\u2005\u00c5] and an oxygen-bound DMSO [Na\u2014O = 2.3165\u2005(12)\u2005\u00c5]. The title complex, [NaRuCl DOI: 10.1107/S1600536811017211/om2427Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Danio rerio ; (ii) the African clawed toad Xenopus laevis ; and iii) the pond snail Radix balthica . Image sequences were analysed using Sparse Optic Flow and the resultant frame-to-frame motion parameters were analysed using Discrete Fourier Transform to quantify the distribution of energy at different frequencies. This spectral frequency dataset was then used to construct a Bray-Curtis similarity matrix and differences in movement patterns between embryos in this matrix were tested for using ANOSIM.Motion analysis is one of the tools available to biologists to extract biologically relevant information from image datasets and has been applied to a diverse range of organisms. The application of motion analysis during early development presents a challenge, as embryos often exhibit complex, subtle and diverse movement patterns. A method of motion analysis able to holistically quantify complex embryonic movements could be a powerful tool for fields such as toxicology and developmental biology to investigate whole organism stress responses. Here we assessed whether motion analysis could be used to distinguish the effects of stressors on three early developmental stages of each of three species: (i) the zebrafish Xenopus laevis at stages 24, 32 and 34 exposed to a salinity of 20, Danio rerio at 33\u2009hpf exposed to 1.5% ethanol, and Radix balthica at stages E4, E9 and E11 exposed to salinities of 5, 10 and 15. This technique was better able to distinguish embryos exposed to stressors than analysis of manual quantification of movement and within species distinguished most of the developmental stages studied in the control treatments.Spectral frequency analysis of these motion parameters was able to distinguish stage-specific effects of environmental stressors in most cases, including This innovative use of motion analysis incorporates data quantifying embryonic movements at a range of frequencies and so provides an holistic analysis of an embryo\u2019s movement patterns. This technique has potential applications for quantifying embryonic responses to environmental stressors such as exposure to pharmaceuticals or pollutants, and also as an automated tool for developmental staging of embryos. The development of imaging technology over the past two decades has enabled significant advances in biology and resulted in an exponential increase in the size and complexity of the data sets generated ,2. HowevAmong the many methods of automated image analysis available to biologists is motion analysis, a technique that has been used for quantifying the behavior of mammals , fish 88, sperm Danio rerio embryos at stages 19\u2009h, 21.5\u2009h and 33\u2009h , and a salinity of 5 : 598 \u2013 298\u2009s; [2]: 200 \u2013 149\u2009s; [3]: 120 \u2013 99\u2009s; [4]: 85 \u2013 54\u2009s; [5]: 49 \u2013 33\u2009s; [6]: 29 \u2013 20.5\u2009s; [7]: 21 \u2013 17\u2009s; [8]: 16 \u2013 15\u2009s; [9]: 14 \u2013 13\u2009s; [10]: 12 \u2013 11\u2009s; [11]: 10 \u2013 5\u2009s; [12]: 4.9 \u2013 3.5\u2009s; [13]: 3.3 \u2013 2.5\u2009s; [14]: 2.4 \u2013 1.7\u2009s; [15]: 1.6 \u2013 1\u2009s; [16]: 0.9 \u2013 0.33\u2009s; [17]: 0.3 \u2013 0.25\u2009s; [18]: 0.24 \u2013 0.1\u2009s. Analysis of Radix balthica\u2019s frame-to-frame motion analysis output was performed using 18 frequency bins for developmental stage comparisons - [1]: 598 \u2013 299\u2009s; [2]: 291 \u2013 145\u2009s; [3]: 120 \u2013 100\u2009s; [4]: 85 \u2013 75\u2009s; [5]: 66 \u2013 60\u2009s; [6]: 54 \u2013 50\u2009s; [7]: 46 \u2013 43\u2009s; [8]: 40 \u2013 37\u2009s; [9]: 35 \u2013 30\u2009s; [10]: 29 \u2013 24\u2009s; [11]: 20 \u2013 17\u2009s; [12]: 15 \u2013 12.5\u2009s; [13]: 12.2 \u2013 10\u2009s; [14]: 9.5 \u2013 5\u2009s; [15]: 2 \u2013 1.1\u2009s; [16]: 1 \u2013 0.68\u2009s; [17]: 0.66 \u2013 0.5\u2009s; [18]: 0.3 \u2013 0.26\u2009s, and 30 frequency bins for environmental stressor analysis - [1]: 300 \u2013 150\u2009s; [2]: 100 \u2013 75\u2009s; [3]: 60 \u2013 50\u2009s; [4]: 43 \u2013 37.5\u2009s; [5]: 33.5 \u2013 30\u2009s; [6]: 27.5 \u2013 25\u2009s; [7]: 23 \u2013 21.5\u2009s; [8]: 20 \u2013 19\u2009s; [9]: 17.5 \u2013 16.5\u2009s; [10]: 16 \u2013 14.5\u2009s; [11]: 13.5 \u2013 12.5\u2009s; [12]: 12 \u2013 11\u2009s; [13]: 10.5 \u2013 9.5\u2009s; [14]: 9 \u2013 6.5\u2009s; [15]: 6.3 \u2013 5\u2009s; [16]: 4.9 \u2013 3.5\u2009s; [17]: 3.3 \u2013 2.5\u2009s; [18]: 2.4 \u2013 2\u2009s; [19]: 1.9 \u2013 1.5\u2009s; [20]: 1.4 \u2013 1\u2009s; [21]: 0.9 \u2013 0.8\u2009s; [22]: 0.7 \u2013 0.65\u2009s; [23]: 0.6 \u2013 0.5\u2009s; [24]: 0.49 \u2013 0.4\u2009s; [25]: 0.39 \u2013 0.34\u2009s; [26]: 0.33 \u2013 0.28\u2009s; [27]: 0.27 \u2013 0.22\u2009s; [28]: 0.21 \u2013 0.18\u2009s; [29]: 0.17 \u2013 0.15\u2009s; [30]: 0.14 \u2013 0.13\u2009s. Radix balthica during its development performs movement types ranging from ciliary driven rotation to muscular driven crawling. As a result, to discern treatments more frequency bins were required for analysis of Radix balthica than for Danio rerio or Xenopus laevis, as movement differences fell within rather than between frequency bins. These extra frequency bins enhanced the frequency discrimination and therefore the detection of subtle changes to these complex movement behaviours.Frame-to-frame motion analysis data were analysed for spectral content using the Discrete Fourier Transform (DFT) . This reThe data resulting from the DFT comprises 72 parameters for each individual. These data were transformed (Log X\u2009+\u20091) and for each species a Bray-Curtis similarity matrix was calculated. This matrix was used to generate Multidimensional Scaling (MDS) plots and ANOSIM was usedThe authors have a patent pending, entitled: Method and system for determining characteristics of an embryo and uses thereof .This work was funded by the Marine Institute at Plymouth University through the HEIF4 programme.TB and OT carried out experimental work and bioimaging. PC carried out programming required for motion analysis and frequency analysis. TB, OT, SDR and JIS participated in the data analysis. All authors read and approved the final manuscript."} +{"text": "Two SnIV atoms are coordin\u00adated by two butyl groups, one benzoate O atom and two bridging O atoms, whereas the other two SnIV atoms are coordinated by two butyl groups, two benzoate O atoms and a bridging O atom. All the butyl groups are equatorial with respect to the SnO3 trigonal plane. In the crystal, mol\u00adecules are linked into a two-dimensional layer parallel to the ab plane by inter\u00admolecular N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and further stabilized by a \u03c0\u2013\u03c0 inter\u00adaction [centroid\u2013centroid distance = 3.6489\u2005(11)\u2005\u00c5]. Intra\u00admolecular N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds stabilize the mol\u00adecular structure. Two of the butyl groups are each disordered over two sets of sites with site-occupancy ratios of 0.510\u2005(4):0.490\u2005(4) and 0.860\u2005(5):0.140\u2005(5).In the title complex, [Sn DOI: 10.1107/S1600536811028212/is2746Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordination geometry around the CuII atom can be described as distorted elongated octa\u00adhedral with Rout = 2.277\u2005(2)\u2005\u00c5, Rin = 2.052\u2005(2)\u2005\u00c5 and a tetra\u00adgonality of 0.9011, acquiring a \u2018static\u2019 stereochemistry. In the supra\u00admolecular network, there are inter\u00admolecular C\u2014H\u22efF and C\u2014H\u22efN inter\u00adactions with R33(16), R22(7), R12(4), R33(16) and C32(7) motifs that lead to an infinite three-dimensional network.In the title compound, [Cu(C DOI: 10.1107/S1600536812028267/ru2037Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The RhI atom is displaced from the plane through these surrounding atoms by 0.0085\u2005(2)\u2005\u00c5. The dihedral angle between the benzene ring and the N\u2014C\u2014C\u2014C\u2014O plane is 89.82\u2005(6)\u00b0, and the N\u2014Rh\u2014O bite angle for the bidentate ligand is 90.53\u2005(6)\u00b0. An inter\u00admolecular C\u2014H\u22efO inter\u00adaction is observed between a methyl group of the benzene ring and a carbonyl O atom.In the title compound, [Rh(C DOI: 10.1107/S1600536812017175/hy2537Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "There are C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.487 \u2005(3)\u2005\u00c5] in the crystal structure. The perchlorate anion is disordered over two positions with an occupancy ratio of 0.628\u2005(9):0.372\u2005(9). In the crystal structure of the title compound, [Cu(HCO DOI: 10.1107/S1600536811039675/jh2326Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The NiII atom is bonded by two O atoms of the bidentate chelating sulfate ligand and the four N atoms of two chelating 2,2\u2032-bi\u00adpyridine ligands. The Ni\u2014N bond lengths range from 2.059\u2005(3) to 2.075\u2005(3)\u2005\u00c5 and the Ni\u2014O bond lengths are 2.098\u2005(3) and 2.123\u2005(3)\u2005\u00c5. The bipyridyl ligands are both close to planar (r.m.s. deviations of 0.254 and 0.0572\u2005\u00c5) and are almost orthogonal, making a dihedral angle of 82.77\u2005(1)\u00b0. In the crystal, the complex and water mol\u00adecules are connected by O\u2014H\u22efO hydrogen bonds. Inter\u00adestingly, six water mol\u00adecules form a chain linking two complex mol\u00adecules via sulfate O atoms. There are also stacking inter\u00adactions between the aromatic rings of neighbouring 2,2\u2032-bi\u00adpyridine ligands with shortest non-covalent contacts of 3.268\u2005(6), 3.393\u2005(6) and 3.435\u2005(5)\u2005\u00c5. One of the three unique water molecules shows half-occupation.The title compound, [Ni(SO DOI: Click here for additional data file.10.1107/S1600536813014219/sj5321Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The negatively charged N atom of the group binds to the terminal SnIV atom at a shorter distance [Sn\u2014N = 2.240\u2005(3)\u2005\u00c5] compared with the neutral N atom that binds to the central SnIV atom [Sn\u2190 N = 2.641\u2005(3)\u2005\u00c5]. The terminal SnIV atom is five-coordinate in a cis-C2SnNO2 trigonal\u2013bipyramidal geometry [C\u2014Sn\u2014C = 127.5\u2005(2)\u00b0], whereas the central SnIV atom is six-coordinate in a C2SnNO3 skew-trazepoidal bipyramidal geometry [C\u2014Sn\u2014C = 145.0\u2005(2)\u00b0].The tetra\u00adnuclear title compound, [Sn DOI: Click here for additional data file.10.1107/S1600536812047708/bt6863Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The RuII atom has a classical three-legged piano-stool environment being coordinated by an \u03b76-benzene ligand [Ru\u2014centroid = 1.6964\u2005(6)\u2005\u00c5], two chloride ligands with an average Ru\u2014Cl bond length of 2.4138\u2005(3)\u2005\u00c5 and a dicyclo\u00adhexyl\u00adphenyl\u00adphosphane ligand [Ru\u2014P = 2.3786\u2005(3)\u2005\u00c5]. The effective cone angle for the phosphane was calculated to be 158\u00b0. In the crystal, weak C\u2014H\u22efCl hydrogen bonds link the RuII complexes into centrosymmetric dimers. The crystal packing exhibits intra- and inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions resulting in a zigzag pattern in the [101] direction.The asymmetric unit of the title compound, [RuCl II\u2013arene complexes, see: Chen et al. (C18H27P)]\u00b71.5C6H6 = 0.018wR(F2) = 0.048S = 1.037589 reflections334 parametersH-atom parameters constrainedmax = 0.54 e \u00c5\u22123\u0394\u03c1min = \u22120.43 e \u00c5\u22123\u0394\u03c1APEX2 global, I. DOI: Click here for additional data file.10.1107/S1600536812044674/cv5349Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the L ligand, the two benzene rings are nearly parallel, forming a dihedral angle of 2.0\u2005(1)\u00b0. In the crystal, inter\u00admolecular O\u2014H\u22efN hydrogen bonds link pairs of mol\u00adecules into centrosymmetric dimers which exhibit \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings [centroid\u2013centroid distance = 3.677\u2005(5)\u2005\u00c5].In the title mol\u00adecule, [V(C DOI: 10.1107/S1600536811010385/cv5063Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CaII atom is octa\u00adcoordinated by two O atoms from two water mol\u00adecules and six O atoms from four acetate ligands. Each acetate acts as a tridentate ligand bridging two CaII atoms, resulting in a chain running along the c axis. O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds connect the chains into a two-dimensional network parallel to [011]. \u03c0\u2013\u03c0 inter\u00adactions between adjacent isoindoline-1,3-dione rings [centroid\u2013centroid distance = 3.4096\u2005(11)\u2005\u00c5] further consolidate the structure. One of the carboxylate O atoms is disordered over two sites in a 0.879\u2005(12):0.121\u2005(12) ratio.In the title complex, [Ca(C N-phthaloylglycine, see: Khan & Ismail 2(H2O)2] = 0.036 wR(F 2) = 0.102 S = 1.04 2339 reflections160 parameters3 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.28 e \u00c5\u22123 \u0394\u03c1min = \u22120.23 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811027851/pv2422Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The complex mol\u00adecules are linked by inter\u00admolecular N\u2014H\u22efO hydrogen bonds into a chain along [010]. \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings and between the pyrazole rings [centroid\u2013centroid distances = 3.772\u2005(2) and 3.546\u2005(2)\u2005\u00c5] connect the chains.In the title complex, [Mn(CH DOI: 10.1107/S1600536811010506/hy2414Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The five Cl atoms bound to Sb [Sb\u2014Cl = 2.4043\u2005(9)\u20132.6262\u2005(11)\u2005\u00c5] form a square-pyramidal coordination environment. In addition, two [SbCl5]2\u2212 anions related by an inversion center are joined by Sb\u22efCl inter\u00adactions [Sb\u22efCl = 3.7273\u2005(14)\u2005\u00c5] into an [Sb2Cl10]4\u2212 dimer with two bridging Cl atoms. The anions, water mol\u00adecules and ammonium groups of the cations are linked by N\u2014H\u22efCl, N\u2014H\u22efO and O\u2014H\u22efCl hydrogen bonds, forming layers parallel to the ac plane. The benzene rings of the 4-methyl\u00adanilinium cations are packed between these layers.The title compound, (C DOI: 10.1107/S1600536812009427/yk2046Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In comparison with the original determination based on photographic data , the current study allows the location of reliable postions for the H atoms and thus leads to better understanding of the inter\u00adionic and inter\u00admolecular inter\u00adactions. The crystal structure consists of an octa\u00adhedrally coordinated cationic CoIII complex ion, an octa\u00adhedrally coordinated anionic CoIII complex ion and a lattice water mol\u00adecule. The complex cation, complex anion and lattice water mol\u00adecule are connected by an intricate network of O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming a three-dimensional structure.The structure of the title compound, [Co(NO al. 1976. Inorg. III complexes, see: Angelici 2(C2H8N2)2][Co(NO2)4(C2H8N2)]\u00b7H2O = 0.035wR(F2) = 0.087S = 1.076280 reflections340 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.72 e \u00c5\u22123\u0394\u03c1min = \u22120.84 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 global, I. DOI: Click here for additional data file.10.1107/S1600536812050325/wm2706Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "One iodo\u00adbenzoate anion and one water mol\u00adecule bridge adjacent Co cations, forming a polymeric chain running along the a axis, while the other iodo\u00adbenzoate anion and water mol\u00adecule coordinate in a monodentate manner to the CoII cation, completing the slightly distorted octa\u00adhedral geometry. In the two independent anionic ligands, the carboxyl\u00adate groups are twisted away from the attached benzene rings by 51.38\u2005(18) and 39.89\u2005(11)\u00b0, and the two benzene rings are nearly perpendicular to each other with a dihedral angle of 86.09\u2005(10)\u00b0. Intra\u00admolecular O\u2014H\u22efO hydrogen bonds between coordinating water mol\u00adecules and adjacent carboxyl\u00adate O atoms help to stabilize the mol\u00adecular structure. In the crystal, weak C\u2014H\u22efO hydrogen bonds link the polymeric chains into a three-dimentional supra\u00admolecular network.The asymmetric unit of the polymeric title compound, [Co(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(H2O)2] = 0.020wR(F2) = 0.051S = 1.204313 reflections224 parameters8 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.60 e \u00c5\u22123\u0394\u03c1min = \u22121.03 e \u00c5\u22123\u0394\u03c1Absolute structure: Flack 1983, 1835 FrFlack parameter: 0.016 (19)APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812015115/xu5504Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "A distorted octahedral coordination geometry of the CoII atom results from ligation of an H atom, which is part of an agostic B\u2014H\u22efCo inter\u00adaction [H\u22efCo = 2.12\u2005(3)\u2005\u00c5], and by five imine N atoms, two from a Bp ligand and three from a Tp ligand. Weak intra- and inter\u00admolecular C\u2014F\u22ef\u03c0 inter\u00adactions with F\u22efcentroid distances ranging from 3.025\u2005(4) to 3.605\u2005(4)\u2005\u00c5 are observed.The title compound, [Co(Cenza 2010. Acta Cr DOI: 10.1107/S1600536811021994/rz2601Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular O\u2014H\u22efO hydrogen bonds lead to the formation of a two-dimensional layer structure parallel to (001). The layers are connected by \u03c0\u2013\u03c0 inter\u00adactions between the pyridyl and benzene rings of the phenanthroline ligands [centroid\u2013centroid distances = 3.591\u2005(1) and 3.610\u2005(1)\u2005\u00c5].In the title mononuclear complex, [Cd(C DOI: 10.1107/S1600536810028175/hy2328Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In both ligands, the benzyl and benzimidazole rings are nearly perpendicular [dihedral angles = 81.7\u2005(2) and 81.5\u2005(2)\u00b0]. The two benzimidazole systems are essentially planar [maximum deviations = 0.015\u2005(3) and 0.020\u2005(2)\u2005\u00c5] and form a dihedral angle of 78.09\u2005(8)\u00b0. In the crystal, centrosymmetrically related mol\u00adecules are linked by pairs of C\u2014H\u22efCl hydrogen bonds into chains parallel to the a axis.In the title compound, [ZnCl DOI: 10.1107/S1600536814002840/rz5104Isup2.hklStructure factors: contains datablock(s) I. DOI: CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The hydrazine carbothio\u00adamide moiety and one of the pyridyl rings together adopt an almost planar arrangement, with a maximum deviation of 0.052\u2005(4)\u2005\u00c5 for the C atom of the thio\u00adurea moiety. There are two mol\u00adecules of methanol solvent per complex in the asymmetric unit. The nonconventional intra\u00admolecular C\u2014H\u22efBr hydrogen bonds make the mol\u00adecule more rigid, whereas the conventional N\u2014H\u22efN and O\u2014H\u22efBr inter\u00admolecular hydrogen-bonding inter\u00adactions, supported with N\u2014H\u22ef\u03c0 inter\u00adactions, establish a supra\u00admolecular linkage among the mol\u00adecules in the crystal. An intermolecular C\u2014H\u22efO inter\u00adaction is also present.In the centrosymmetric binuclear title compound, [Cu DOI: 10.1107/S1600536812005934/fj2511Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Within the complex, the carb\u00adoxy\u00adlic acid forms intra\u00admolecular O\u2014H\u22efO hydrogen bonds, while the mol\u00adecules are assembled through N\u2014H\u22efO(carbox\u00adyl) hydrogen bonds into chains extending along the a-axis direction. These chains are further linked by weak \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid separation = 3.930\u2005(2)\u2005\u00c5].The structure of the title compound, [Cu(C DOI: 10.1107/S160053681102839X/zs2128Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angle between the benzene rings is 12.27\u2005(11)\u00b0. Adjacent complex mol\u00adecules are linked into dimers by O\u2014H\u22efO hydrogen bonds, generating rings of R 1 2(6) and R 1 2(5) graph-set motifs, and by aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions, with a centroid\u2013centroid distance of 3.812\u2005(2)\u2005\u00c5. The crystal packing is further stabilized by inter\u00admolecular C\u2014H\u22efN hydrogen bonds. The C and N atoms of the acetonitrile solvent mol\u00adecule are located on a crystallographic twofold axis.In the mononuclear complex mol\u00adecule of the title compound, [Cr(C DOI: 10.1107/S1600536811037275/rz2636Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular O\u2014H\u22efI hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings [centroid\u2013centroid distance = 3.790\u2005(2)\u2005\u00c5] are present in the crystal structure.In the centrosymmetric dinuclear title compound, [Cd DOI: 10.1107/S1600536810041681/hy2362Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The benzene and pyridine rings are oriented at a dihedral angle of 56.7\u2005(6)\u00b0. The crystal packing is stabilized by C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings [centroid\u2013centroid distance = 3.796\u2005(6)\u2005\u00c5].In the centrosymmetric binuclear molecule of the title complex, [Hg DOI: 10.1107/S1600536812039050/hy2585Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In addition, there is a Zn\u22efS contact involving a symmetry-related S atom which, when considered, forms a pseudo-square-pyramidal coordination with respect to the ZnII ion. Three of the ethyl groups are disordered over two sites with occupancy ratios of 0.841\u2005(10):0.159\u2005(10), 0.802\u2005(10):0.198\u2005(10) and 0.457\u2005(13):0.543\u2005(13). Weak intra\u00admolecular C\u2014H\u22efN and C\u2014H\u22efS inter\u00adactions contribute to the stability of the mol\u00adecular conformation. Inter\u00admolecular C\u2014H\u22efS contacts, weak C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centriod distances = 3.832\u2005(4) and 3.850\u2005(5)\u2005\u00c5] contribute to the stabilization of the crystal structure.In the title compound, [Zn(C DOI: 10.1107/S1600536810028588/lh5086Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII atom together with its four coordinated O atoms are in a distorted planar square arrangement while the nitro\u00adgen and the other CuII atom are located in apical positions. The whole mol\u00adecule looks like a paddle-wheel. In the crystal, chains are assembled along the b axis through C\u2014H\u22efO hydrogen bonds and slipped \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of neighbouring mol\u00adecules [centroid\u2013centroid distance = 3.6929\u2005(3)\u2005\u00c5 and slippage = 0.641\u2005(1)\u2005\u00c5].In the binuclear copper(II) title complex, [Cu II carboxyl\u00adato compounds with subnormal magnetic moments, see: Kato et al. 4(C2H3N)2] = 0.037wR(F2) = 0.105S = 1.073466 reflections271 parametersH-atom parameters constrainedmax = 0.55 e \u00c5\u22123\u0394\u03c1min = \u22120.32 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536812049410/lr2089Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, the complex anions and cations are linked via N\u2014H\u22efCl hydrogen bonds into layers parallel to (101). Short Br\u22efCl contacts of 3.4239\u2005(11) and 3.4503\u2005(12)\u2005\u00c5 are observed, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine and pyridinium rings, with alternating centroid-to-centroid distances of 3.653\u2005(2) and 3.845\u2005(2)\u2005\u00c5.The structure of the title salt, (C DOI: Click here for additional data file.10.1107/S1600536813011884/wm2739Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The structure contains two uncoordinated water mol\u00adecules. In the crystal, N\u2014H\u22efO, O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings of the pydc ligands, with a centroid\u2013centroid distance of 3.4582\u2005(18)\u2005\u00c5, stabilize the structure.In the title compound, (C DOI: 10.1107/S1600536811024366/hy2434Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angles formed between the mean planes of the pyrazole rings and the phenanthroline system are 15.74\u2005(15) and 16.30\u2005(13)\u00b0. In the crystal, there is a \u03c0\u2013\u03c0 stacking inter\u00adaction involving two symmetry-related pyrazole rings, with a centroid\u2013centroid distance of 3.664\u2005(3)\u2005\u00c5. In addition, there is a relatively short inter\u00admolecular contact between C atoms [C\u22efC = 3.399\u2005(6)\u2005\u00c5] involving symmetry-related pyridine rings along the a axis.In the title complex, [Cd(NCS) DOI: 10.1107/S1600536810051275/lh5181Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The o-nitro\u00adsotoluene ligand is N-bound to iron(II) [Fe\u2014N = 1.8406\u2005(18)\u00c5 and Fe\u2014N\u2014O = 122.54\u2005(14)\u00b0]. The axial N\u2014Fe\u2014N linkage is almost linear, with a bond angle of 177.15\u2005(7)\u00b0. One phenyl group of the porphyrin ligand is disordered over two orientations in a 0.710\u2005(3):0.290\u2005(3) ratio. The di\u00adchloro\u00admethane solvent mol\u00adecule was severely disordered and its contribution to the scattering was removed with the SQUEEZE routine .The solvated title compound, [Fe(CSpek 1990. Acta Cr DOI: 10.1107/S160053681400083X/hb7178Isup2.hklStructure factors: contains datablock(s) I. DOI: CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II ion in the title complex, [PtBr2(C10H9N3)], is four-coordinated in an essentially square-planar environment by two N atoms from a chelating di-2-pyridyl\u00adamine (dpa) ligand and two Br\u2212 anions. The dpa ligand is not planar, with the dihedral angle between the pyridine rings being 40.8\u2005(2)\u00b0. The complex mol\u00adecules are stacked in columns along [001] through \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings [centroid\u2013centroid distances = 3.437\u2005(3) and 3.520\u2005(3)\u2005\u00c5]. Inter\u00admolecular N\u2014H\u22efBr hydrogen bonds connect the mol\u00adecules into chains running along [010]. Intra\u00admolecular C\u2014H\u22efBr interactions are also observed.The Pt II complex [PtCl2(dpa)], see: Li & Liu ] = 0.029wR(F2) = 0.079S = 1.102973 reflections145 parametersH-atom parameters constrainedmax = 1.41 e \u00c5\u22123\u0394\u03c1min = \u22122.22 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The amine N\u2014H group and pyridine N atom are syn allowing for the formation of centrosymmetric eight-membered {\u22efHNCN}2 synthons via N\u2014H\u22efN hydrogen bonds. The two-mol\u00adecule aggregates are sustained in the three-dimensional crystal packing via C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance for pyridyl rings = 3.7535\u2005(12)\u2005\u00c5]The title amine, C DOI: 10.1107/S1600536811044059/hg5117Isup2.hkl Structure factors: contains datablock(s) I. DOI: 10.1107/S1600536811044059/hg5117Isup3.cml Supplementary material file. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In each mol\u00adecule, the CuII ion is five-coordinated in a distorted square-pyramidal geometry wherein the basal plane is defined by the N,N\u2032,N\"-tridentate Schiff base and one N-bound thio\u00adcyanate ligand. The second N-donor thio\u00adcyanate group, located at the apical site, completes the coordination environment. In the crystal, inter\u00admolecular C\u2014H\u22efS and C\u2014H\u22efN hydrogen bonds link adjacent mol\u00adecules into infinite layers parallel to the ac plane. Intra\u00admolecular C\u2014H\u22efN inter\u00adactions are also observed.The asymmetric unit of the title compound, [Cu(NCS) For the al. 2011. For a d al. 1984. 2(C11H17N3)] = 0.045 wR(F 2) = 0.128 S = 1.00 7159 reflections385 parametersH-atom parameters constrainedmax = 1.04 e \u00c5\u22123 \u0394\u03c1min = \u22120.81 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811022057/is2717Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In complex A, [Zn2(C8H7O2)4(H2O)2], each Zn cation is chelated by a 4-methyl\u00adbenzoate (PMB) anion and coordinated by a water mol\u00adecule, and is further bridged by two PMB anions in a trigonal-bipyramidal geometry. In complex B, [Zn2(C8H7O2)4(C6H6N2O)2], each ZnII cation is coordinated by a monodentate PMB anion and one nicotinamide (NA) ligand, and is further bridged by two PMB anions in a tetra\u00adhedral geometry. Weak intra-mol\u00adecular \u03c0\u2013\u03c0 stacking between adjacent benzene rings is observed in complex B, the centroid\u2013centroid distance being 3.710\u2005(2)\u2005\u00c5. Extensive O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonding and weak C\u2014H\u22efO hydrogen bonding is present in the crystal structure. The crystal studied was a racemic twin; the minor twin component refined to 38%.The crystal structure of the title compound, [Zn DOI: 10.1107/S1600536810022476/xu2762Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "AbstractHarpapion safranumsp. n. and Harpapion borisisp. n. are described and figured. Harpapion vietnamense is recorded as new for China. The genitalia andterminalia of H. considerandum, H. coelebs andH. vietnamense are redescribed and redrawn. The diagnostic characters of Harpapion are defined. A key to the known species of the genus Harpapion from China is provided. Affinities with the genus Flavopodapion Korotyaev, 1987 are discussed. Harpapion was erected and initially placed as a subgenus of Apiotherium Beguin-Billecocq, 1905 by Apioninaeincertae sedis by Aspidapiini Alonso-Zarazaga, 1990 based on its elongate-triangular scutellum and the genitalia with apparently primitive traits and Harpapion vietnamense , respectively from China and Vietnam were identified as new records of Harpapion in Southeast Asia , and then Harpapion compared with Pseudaspidapion Wanat, 1990.The apionid genus e traits . Moreovee traits until twast Asia . Wanat , Harpapion coelebs andHarpapion vietnamense.Recently, we obtained two more specimens of the monotypic species Institute of Zoology, Chinese Academy of Sciences, Beijing (IZCAS) and the Biological Museum, Sun Yat-Sen University, Guangdong (BMSYU). Type and identified specimens were obtained from BMSYU, Natural History Museum (NHM), Zoological Institute (ZIN) on loan or belong to IZCAS.Materials examined of the new species for this study are to be deposited in the Descriptions were made and photographs were taken with a Canon EOS 5D mounted on a Nikon SMZ1500. Extended focus images were generated with CombineZM and edited with Adobe Photoshop CS 5.0 when required. Microscopic slides were studied under a Leica DM 2500 microscope and photos were taken with a Nikon CoolPix P7100. Drawings were made from the original photographs by using the software Adobe Illustrator CS5.0, or directly by using a drawing tube attached to the microscope.Scanning Electron Microscope (SEM) photos were taken with a FEI ESEM Quanta 450 device and the software xT microscope control. Specimens were cleaned by hair pencil and mounted on the mounting card directly.Nomenclature of the rostral parts follows The dissecting method used follows After description, the genitalia and other parts of each specimen were placed in DMHF on an acetate card for long term conservation .Labels are described as they are (in Chinese or Cyrillic), with pinyin romanization for Chinese and ISO 9:1995 for Russian, and translations in square brackets; labels are separated by semicolons and lines by slashes. Alternative modern pinyin romanizations are also given in some cases where labels are written using other romanization systems, like Wade-Giles.Harpapion could be defined as follows: 1) scutellum triangular, distinctly elongate and basally projecting, apically raised (Pseudaspidapion) the pygidium is of the aspidapionine type, subsemicircular in dorsal view, with the apical flange strongly raised, the transverse sulcus deep but not cutting the sides; 8) the ninth sternite is Y-shaped, slender and subsymmetrical, not winged; 9) the apex of the penis is moderately to distinctly curved in lateral view, sometimes the pedon is recurved at apex; 10) the tegminal plate is articulated with the free ring, laterally developed, enveloping; the parameroid lobes present a usually well developed apical membranous area with a small apical notch, the basal sclerotized area has a medial sinuation in its front margin and bears 4-7 macrochaetae on each side; the fenestrae are present and variable; the prostegium is bidentate with two lateral projections (absent in Harpapion coelebs).According to species examined, the diagnostic characters of y raised ; 2) the idapion) , 16; 3) http://species-id.net/wiki/Harpapion_considerandumPygidium subsemicircular in dorsal view, 0.83\u00d7 as long as wide, apical flange strongly raised, transverse sulcus deep Y-shaped, slender and nearly symmetrical, not winged, manubrium about 1.21\u00d7 as long as arms . Penis i1\u2642: : Magila / E. Africa / A. V. Legros / 98-190; : Wagner det. \u2642 / considerandum F\u00e5hr.; 1\u2640: : X. 1983 / in clove tree / CIEA15888; : Tanzania / Zanjibar / Kitunda; : pres by: Comm Inst Ent B. M. 1984-1. Specimens in the NHM.Harpapion considerandum s. str. has been recorded from the following African countries: Guinea, Cameroon, Republic of the Congo, Democratic Republic of the Congo (ex Zaire), Chad, Ethiopia, Kenya, Tanzania, Uganda, Angola, Zimbabwe and South Africa from Senegal, Guinea, Mali, Nigeria and Tanzania from Tanzania and Angola . Two subTanzania ; and sspd Angola .http://species-id.net/wiki/Harpapion_coelebsPygidium subsemicircular in dorsal view, 0.83\u00d7 as long as wide, apical flange strongly raised, transverse sulcus distinctly depressed and wide, disc with 2\u20133 rows of punctures between disc and sulcus, pubescent sparse and minute : \u4e91\u5357\u666f\u4e1c\u8463\u5bb6\u575f [Y\u00fann\u00e1n J\u01d0ngD\u014dng D\u01d2ngji\u0101f\u00e9n] / 1956.V.28/1250m / leg. \u514b\u96f7\u8ba9\u8bfa\u592b\u65af\u57fa [Kryzhanovsky]; : \u042e\u043d\u044c\u043d\u0430\u043d\u044c, 10 \u043a\u043c. N \u0426\u0437\u0438\u043d- / \u0434\u0443\u043d\u0430 [-D\u014dng], 1250 \u043c, 28. V. 1956 / \u041a\u0440\u044b\u0436\u0430\u043d\u043e\u0432\u0441\u043a\u0438\u0439 [Kryzhanovsky].Yunnan.http://species-id.net/wiki/Harpapion_vietnamensePygidium subsemicircular in dorsal view, 0.89\u00d7 as long as wide, apical flange strongly raised, transverse sulcus deep and narrow, disc nearly smooth, without visible punctures or pubescence ] / \u91c7\u96c6\u4eba\uff1a\u5b5f\u4ee4\u66fe [C\u01ceij\u00edr\u00e9n (leg.): M\u00e8ng L\u00ecngz\u0113ng]; Guomenshan, VI/1D / 06.05.2009/ leg. L. Z. Meng; IOZ(E)1369311; 1\u2642: \u4e91\u78a7\u5927\u5f00\u6cb3 [Y\u00fan B\u00ec D\u00e0k\u0101ih\u00e9] / 1957.IV.23 / \u6731\u589e\u6d69 [leg. Zh\u016b Z\u0113ngh\u00e0o]; IOZ(E)1639312.Holotype. \u2642 : \u0412\u044c\u0435\u0442\u043d\u0430\u043c, \u043f\u0440\u043e\u0432. \u0425\u0430\u0448\u043e\u043d\u0431\u0438\u043d\u044c / 7 \u043a\u043c \u044e.-\u0432. \u0425\u043e\u0430\u0431\u0438\u043d\u044f, \u0432\u0442\u043e\u0440\u0438\u0447\u043d\u044b\u0439 \u0442\u0440\u043e\u043f\u0438\u0447\u0435\u0441\u043a\u0438\u0439 \u043b\u0435\u0441 \u0438 \u043a\u0443\u0441\u0442\u0430\u0440\u043d\u0438\u043a \u043d\u0430 \u0441\u043a\u043b\u043e\u043d\u0435 / 17. X 1976 / leg. \u041b. \u041d. \u041c\u0435\u0434\u0432\u0435\u0434\u0435\u0432 [L. N. Medvedev]; 1\u2642: \u4e91\u5357\u666f\u6d2a\u52d0\u6d77\u53bf\u7eb3\u677f\u6cb3 [Y\u00fann\u00e1n J\u01d0ngh\u00f3ng M\u011bngh\u01ceixi\u00e0n N\u00e0b\u01cenh\u00e9] / \u4fdd\u62a4\u533a\u8fc7\u95e8\u5c71\uff08\u68ee\u6797\uff09 [b\u01ceoh\u00f9q\u016b Gu\u00f2m\u00e9nsh\u0101n (s\u0113n l\u00edn [forest]) / 2009.V.06, 1114m; Yunnan (new record for China), Vietnam.Wang & Alonso-Zarazagasp. n.http://zoobank.org/887BB3CE-2FF2-4337-A439-88763EA93AE1http://species-id.net/wiki/Harpapion_safranumMeasurements (in mm): Standard length: 1.84. Rostrum: length: 0.77, maximum width: 0.16. Pronotum: median length: 0.53, maximum width: 0.57. Elytra: median length: 1.33, maximum width: 0.92.(holotype). Integument and grayish acute hairs on antennae, tibiae and tarsomeres. Pronotal vestiture centripetal, scales on apex parallel to margin, but on base perpendicular to margin, pronotal disc with scales distinctly longer and thicker than on legs, reaching base of anterior scales. Elytral scales in one row per interstriae, two irregular rows on disc, scales on striae 1/2-2/3 as long as scales on interstriae. One specialized seta on apical region of 9th interstria.Rostrum cylindrical and moderately robust, in dorsal view 8.25\u00d7 as long as apical width, 1.45\u00d7 as long as pronotum in midline, widest at mesorostrum, prorostrum constricted apicad, metarostrum with sides almost parallel, metarostrum with median dorsal area impunctate, dorsal submedial sulci and dorsal submedial keels weak, minutely punctate and pubescent, lateral area of metarostrum and prorostrum with weak oblong confluent punctures, weakly microreticulate, apical third of prorostrum almost impunctate, smooth and shining; in lateral view moderately curved, sides converging to apex, carinae and sulci weak, ventral medial keel fine and complete, ventral sublateral keels with dense line of scales under mesorostrum.Head transverse, frons very weakly convex, as wide as metarostrum, constricted behind eyes, medial area impunctate and glabrous, lateral areas with irregular rows of punctures and scales, subocular keel not reaching middle of eyes, area between subocular keel flat, microreticulate and impunctate. Eyes round, moderately convex.Antennae inserted at basal 0.23 of rostral length, scape 3.20\u00d7 as long as wide, about as long as mesorostral width. Pedicel 2.00\u00d7 as long as wide, as long as desmomeres 2+3, desmomeres 2\u20133 1.33\u00d7 as long as wide, desmomeres 4\u20135 and 7 1.00\u00d7 as long as wide, desmomere 6 0.75\u00d7 as long as wide. Club oval, slightly flattened, 1.88\u00d7 as long as wide, as long as last 5.5 desmomeres, sutures obsolete.Pronotum campaniform, 0.93\u00d7 as long as wide, apical constriction relatively strong, little wider at base than at middle, base 1.39\u00d7 as wide as apex, bisinuate with rounded medial projection towards scutellum, basal flange developed. Prescutellar fovea shallow, very short, about as broad as diameter of one puncture, as long as 2\u20133 diameters, reaching 1/4 of pronotum. Discal punctures relatively deep, ca. 0.5\u20131\u00d7 diameter apart, interspaces moderately convex, microreticulate.Scutellum elongate, triangular, ca. 2.00\u00d7 as long as wide, 2 basal tubercles fused at base in front view, apical tubercle rounded, slightly prominent and hardly visible in lateral view : Standard length: 1.62\u20131.90. Rostrum: length: 0.61\u20130.75, maximum width: 0.14\u20130.16. Pronotum: median length: 0.47\u20130.57, maximum width: 0.51\u20130.60. Elytra: median length: 1.26\u20131.46, maximum width: 0.72\u20130.92. Female paratypes 1639313; Paratypes: 3\u2642: Kwangtung [Gu\u01cengd\u014dng] S. China / Loh Fau Shan, [Lu\u00f3f\u00fash\u0101n], / Poh-lo [B\u00f3lu\u00f3] District / April 6\u20138, 1934 / K. C. Yeung; En-071380~En-071382; 1\u2642: Hong Kong: Un-long [Yu\u00e1nl\u00e1ng], / New Territories / September 19. 1940 / J. Linsley Gressitt; En-071403; 1\u2642: Hainan I. South China / Ta-hian [probably D\u00e0oxi\u01ceng], Alt. 300 met. / N. side, 5- Finger Mts. [W\u00fazh\u012dsh\u0101n] / VI-12-1935 / L. Gressitt / En-071423; 1\u2640: Kwangtung [Gu\u01cengd\u014dng], S. China / Ho-y\u00fcn [Heyun] to Wui-lung [probably W\u00e9il\u00f3ng] / Ho-yuen [H\u00e9yu\u00e1n] District / Apr. 7, 1940. J. L. / Gressitt & F. K. To; En-071409; 1\u2640: Kwangtung [Gu\u01cengd\u014dng], S. China. / Sin-fung [X\u012bnf\u0113ng] to Lung-kai [Longgai (probably L\u00f3ngj\u012be which can be found on the modern maps)]. / Sin-fung [X\u012bnf\u0113ng] & Lien-p\u2019ing [Li\u00e1np\u00edng] / Dist\u2019s, Apr. 12, 1940 / L. Gressitt & F. E. To; En-071367.Guangdong, Hong Kong, Hainan.Holotype will be deposited in IZCAS, while all paratypes in BMSYU.Harpapion safranum sp. n., can be easily recognized from other species from China by its external characters However, it is extremely similar to Harpapion considerandum which can be distinguished from the former by genitalia which were described above and illustrated in safranum after its testaceous legs. This is a Medieval Latin name of the plant now called Crocus sativus L. (saffron) which yields a yellowish-orange dye. It is considered a noun in apposition.This species is named Wang & Alonso-Zarazagasp. n.http://zoobank.org/03462889-EF2B-418A-BAE5-394758F1E2DBhttp://species-id.net/wiki/Harpapion_borisiMeasurements (in mm). Standard length: 1.22. Rostrum: length: 0.44, maximum width: 0.08. Pronotum: median length: 0.36, maximum width: 0.41. Elytra: length: 0.94, maximum width: 0.54.(holotype). Integument generally piceous, tibiae and tarsi relatively paler and antennae pale reddish brown ] / \u91c7\u96c6\u4eba\uff1a\u5b5f\u4ee4\u66fe [C\u01ceij\u00edr\u00e9n (leg.): M\u00e8ng L\u00ecngz\u0113ng]; Guomenshan VI/1D / 16.03.2009 / leg. L. Z. Meng; IOZ(E)1639309; Paratype: 1\u2642, : \u4e91\u5357\u666f\u6d2a\u52d0\u6d77\u53bf\u7eb3\u677f\u6cb3 [Y\u00fann\u00e1n J\u01d0ngh\u00f3ng M\u011bngh\u01ceixi\u00e0n N\u00e0b\u01cenh\u00e9] / \u4fdd\u62a4\u533a\u8fc7\u95e8\u5c71\uff08\u68ee\u6797\uff09 [b\u01ceoh\u00f9q\u016b Gu\u00f2m\u00e9nsh\u0101n (s\u0113n l\u00edn [forest])] / 2009.VI.26 1114m; 22.24644\u00b0N, 100.60610\u00b0E\u98de\u963b [F\u0113iz\u016d (flight intercept)] / \u91c7\u96c6\u4eba\uff1a\u5b5f\u4ee4\u66fe [C\u01ceij\u00edr\u00e9n (leg.): M\u00e8ng L\u00ecngz\u0113ng]; Guomenshan VI/1D / 26.06.2009 / leg. L. Z. Meng; IOZ(E)1639310.Holotype: 1\u2642: : \u4e91\u5357\u666f\u6d2a\u52d0\u6d77\u53bf\u7eb3\u677f\u6cb3 [Y\u00fann\u00e1n J\u01d0ngh\u00f3ng M\u011bngh\u01ceixi\u00e0n N\u00e0b\u01cenh\u00e9] / \u4fdd\u62a4\u533a\u8fc7\u95e8\u5c71\uff08\u68ee\u6797\uff09 [b\u01ceoh\u00f9q\u016b Gu\u00f2m\u00e9nsh\u0101n (s\u0113n l\u00edn [forest])] / 2009.III.16 1114m; Yunnan.Both holotype and paratype will be deposited in IZCAS.Harpapion borisi sp. n. can be distinguished from other congeners by the following traits: 1) body standard length 1.22\u20131.24 mm (the others more than 1.5 mm); 2) elytral scales with similar size, in one regular row per interstria; 3) rostral pubescence not surpassing middle of prorostrum, nearly entire prorostrum glabrous; 4) prostegial teeth short and narrow; 5) tegminal median unsclerotized strip absent; 6) spiculum gastrale with manubrium about as long as arms.Apionidae from South China and helped us in many ways.This species is named after the Russian curculionidologist Boris A. Korotyaev, who has much improved the taxonomy of Harpapion considerandum is short and medially sinuate, and the apical membranous area is wellFlavopodapion Korotyaev, 1987 ) with the manubrium distinctly shorter than the arms ) resemblender SEM . On the arpapion . Other carpapion coincidethe arms , the parthe arms . However"} +{"text": "In the crystal, mol\u00adecules are assembled by C\u2014H\u22efO hydrogen bonding and \u03c0\u2013\u03c0 inter\u00adactions between bipy groups [centroid\u2013centroid distances = 3.7686\u2005(16) and 3.7002\u2005(16)\u2005\u00c5] into a three-dimensional network. The nitrite anion is equally disordered over two sets of sites.In the title compound, [Cu(NO DOI: 10.1107/S1600536811002571/kp2303Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII ion is coordinated by two N and four O atoms from one bis\u00ad{[bis\u00ad(carboxyl\u00adatometh\u00adyl)amino]\u00admeth\u00adyl}phosphinate ligand in a distorted octa\u00adhedral coordination geometry. The two crystallographically independent potassium ions exhibit different coordination environments. The potassium ion in a general position is hepta\u00adcoordinated by five carboxyl\u00adate O atoms, one phosphinate O atom and one water mol\u00adecule [K\u2014O = 2.718\u2005(3)\u20133.040\u2005(3)\u2005\u00c5], and the potassium ion situated on the twofold rotation axis is hexa\u00adcoordinated by four carboxyl\u00adate O atoms and two water mol\u00adecules [K\u2014O = 2.618\u2005(3)\u20132.771\u2005(3)\u2005\u00c5]. The water mol\u00adecules are also involved in formation of inter\u00admolecular O\u2014H\u22efO hydrogen bonds.In the title compound, K DOI: 10.1107/S1600536811053608/cv5214Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "O\u2014H\u22efO hydrogen bonding between \u2013COOH and \u2013COO\u2212 groups of adjacent mol\u00adecules and between carboxyl\u00adate groups and coordinated and uncoordin\u00adated water mol\u00adecules leads to a three-dimensional structure which is further stabilized by weak \u03c0\u2013\u03c0 inter\u00adactions of adjacent phen ligands with centroid\u2013centroid separations of 4.2932\u2005(1)\u2005\u00c5.In the title complex, [Mn(C DOI: 10.1107/S1600536810048142/wm2428Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The thia\u00adzolidine ring mean plane makes a dihedral angle of 59.08\u2005(11)\u00b0 with the pyrrolidine ring mean plane, which in turn makes a dihedral angle of 83.40\u2005(10)\u00b0 with the cyclo\u00adpentane ring, indicating that the latter two rings are almost orthogonal to one another. In the crystal, a pair of C\u2014H\u22efO hydrogen bonds link the mol\u00adecules forming inversion dimers. The dimers are linked via \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.7764\u2005(10)\u2005\u00c5] involving the quinoxaline moieties forming chains propagating along [1-10].In the title compound, [Fe(C DOI: 10.1107/S1600536813022605/su2630Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Cl atom of the mol\u00adecule with the smaller C\u2014Sn\u2014C angle inter\u00adacts weakly with the SnIV atom of the mol\u00adecule with the wider C\u2014Sn\u2014C angle at an Sn\u22efCl distance of 3.595\u2005(1)\u2005\u00c5. Weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonding is present in the crystal structure.The asymmetric unit of the title complex, [Sn(CH DOI: 10.1107/S1600536810027790/xu2800Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordination number of copper is six with four basal O atoms, one axial N atom and one axial Cu atom. Four acetate ligands act as bidentate linker and connect two Cu atoms, with a crystallographic inversion center located at the mid-point of the Cu\u2014Cu bond. The acetate ligands form slightly distorted square planes around each metal ion, while the copper ions are displaced by 0.2089\u2005(4)\u2005\u00c5 from these planes towards the N atoms. Thus, the Cu\u2014Cu distance is elongated to 2.6378\u2005(7)\u2005\u00c5, compared with the 2.2180\u2005(7)\u2005\u00c5 distance between the two basal planes. The angle between the basal plane and the Cu\u2014N bond is 4.84\u2005(6)\u00b0.The title compound, [Cu LCu)2(\u03bc-O2CCH3)4 complexes is the crystal structure of cupric acetate monohydrate (L = water), see: Van Niekerk & Schoening L = 2-amino-benzothia\u00adzole, see: Sun et al. L = benzothia\u00adzole, see: Ford et al. 4(C7H5NS)2] = 0.032 wR(F 2) = 0.075 S = 0.97 2784 reflections165 parametersH-atom parameters constrainedmax = 0.45 e \u00c5\u22123 \u0394\u03c1min = \u22120.45 e \u00c5\u22123 \u0394\u03c1 X-AREA used to solve structure: SIR92 I, global. DOI: 10.1107/S1600536811027140/hg5042Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atom in the title complex, [HgI2(C12H6N2O2)], is tetra\u00adhedrally coordinated by the N atoms of the chelating 1,10-phenanthroline-5,6-dione ligand and two I atoms. The range of tetra\u00adhedral angles is broad, viz. 68.94\u2005(17)\u00b0 for the chelate angle to a wide 132.627\u2005(15)\u00b0 for the I\u2014Hg\u2014I angle. The ligand mol\u00adecule is non-planar with the O atoms lying 0.422\u2005(5) and \u22120.325\u2005(5)\u2005\u00c5 out of the plane through the remaining atoms [r.m.s. deviation = 0.068\u2005\u00c5]. Mol\u00adecules are consolidated in the crystal packing by C\u2014H\u22efO inter\u00adactions.The Hg DOI: 10.1107/S1600536811038748/hg5098Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "A twofold rotation axis passes through the Fe4S4 core, completing the coordination of the four Fe atoms with two penta\u00admethyl\u00adcyclo\u00adpenta\u00addienyl ligands and two chelating dithiol\u00adate ligands. There are three short Fe\u2014Fe and three long Fe\u22efFe contacts in the Fe4S4 core, suggesting bonding and non-bonding inter\u00adactions, respectively. The Fe\u2014S bonds in the Fe4S4 core range from 2.1523\u2005(5) to 2.2667\u2005(6)\u2005\u00c5 and are somewhat longer than the Fe\u2014S bonds involving the dithiol\u00adate ligand.The title compound, [Fe DOI: Click here for additional data file.10.1107/S1600536813005564/wm2725Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The SnIV atom is displaced out of the C3Sn girdle of the trans-C3SnO2 trigonal-bipyramidal polyhedron in the direction of the covalently-bonded O atom [Sn\u2014O\u2014C = 137.63\u2005(11)\u00b0] by 0.247\u2005(1)\u2005\u00c5; the geometry is distorted towards an octa\u00adhedron by a remote O atom of the meth\u00adoxy subsituent [Sn\u22efO = 3.019\u2005(1)\u2005\u00c5]The formyl\u00admeth\u00adoxy\u00adnitro\u00adphenoxide ions in the polymeric title compound, [Sn(C DOI: 10.1107/S1600536811016436/jh2283Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CoII atoms are bridged by two carboxyl\u00adate O atoms from two DNSA ligands, forming a centrosymmetric dinuclear structure. Neighbouring dinuclear units inter\u00adact with each other through two types of \u03c0\u2013\u03c0 inter\u00adactions between the L ligands [shortest centroid\u2013centroid distance = 3.646\u2005(3)\u2005\u00c5] and between the L and DNSA ligands [shortest atom-to-centroid distance = 3.794\u2005(3)\u2005\u00c5]. N\u2014H\u22efO, O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds are observed, which lead to a three-dimensional structure.In the title compound, [Co DOI: 10.1107/S1600536810017629/hy2303Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The SnIV atom shows cis-C2SnCl2O trigonal\u2013bipyramidal coordination [C\u2014Sn\u2014C = 157.0\u2005(1)\u00b0]; however, two [Me2SnCl2(H2O)] units are linked by a tin\u2013chlorine bridge [Sn\u2190Cl = 3.247\u2005(1)\u2005\u00c5] across a center of inversion, generating a dinuclear species, so that the geometry is better regarded as a mer-C2SnCl3O octa\u00adhedron. The crown ether inter\u00adacts through O\u2014H\u22efO hydrogen with the metal atom through the coordinated water mol\u00adecules in an outer-sphere manner, generating a hydrogen-bonded chain running along [101]. The 15-crown-5 mol\u00adecule is disordered over the 2/m site.The Sn, Cl and water O atoms of the title compound, [Sn DOI: 10.1107/S1600536812008781/xu5444Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The conformation of the aliphatic C\u2014C\u2014C\u2014C chain is gauche [torsion angle = \u221267.7\u2005(8)\u00b0]. A weak C\u2014H\u22efBr inter\u00adaction helps to establish the conformation. In the crystal, there is a weak secondary bonding inter\u00adaction [Te\u22efN = 3.456\u2005(11)\u2005\u00c5] between the Te atom and the N atom of the solvent mol\u00adecule, which completes a distorted TeNCBr4 octa\u00adhedron. Inversion dimers linked by pairs of C\u2014H\u22efBr inter\u00adactions are also observed.In the title compound, [TeBr DOI: Click here for additional data file.10.1107/S1600536812051707/hb7012Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The [SbBr6]3\u2212 and [SbBr4]\u2212 anions are linked together by two long Sb\u2014Br bonds of 3.2709\u2005(8) and 3.5447\u2005(7)\u2005\u00c5 into {[Sb2Br10]4\u2212}n chains along [001]. One of the three tetra\u00admethyl\u00adethylendi\u00adammonium cations is disordered and was refined using a split model (occupancy ratio 0.58:0.42). The cations and the water mol\u00adecule are connected to the {[Sb2Br10]4\u2212}n polymeric anions by weak N\u2014H \u22efBr and O(water)\u2014H \u22efBr hydrogen bonding.The asymmetric unit of the title compound {(C DOI: 10.1107/S1600536813028894/nc2318Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The azotetra\u00adzolate ligand displays a bridging coordination mode, forming an infinite zigzag chain. Inter\u00admolecular O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonding and offset face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.4738\u2005(13)\u2005\u00c5] lead to a three-dimensional network.In the title compound, {[Fe(C DOI: 10.1107/S1600536810039632/ng5036Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the complex cation [Mn2(C2O4)(C10N2H8)4]2+, two MnII atoms are bridged by a bis\u00ad(bidentate) oxalate ligand, each MnII atom being further coordinated by two bpy ligands in a distorted octa\u00adhedral geometry. The distance between the two six-coordinated metal atoms is 5.583\u2005(1)\u2005\u00c5. \u03c0\u2013\u03c0 stacking inter\u00adactions [inter\u00adplanar distances between bpy rings = 3.739\u2005(1)\u2005\u00c5] are essential to the supramolecular assembly. There are extensive inter\u00adionic C\u2014H\u22efO inter\u00adactions between the cations and between the cation and anion. Three of the four perchlorate O atoms are disordered over two sets of sites with occupancy ratios of 0.852\u2005(6):0.148\u2005(6).The unit cell of the title compound, [Mn DOI: 10.1107/S1600536811038475/bv2191Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the complex, there is an intra\u00admolecular N\u2014H\u22efN hydrogen bond. In the crystal, the binuclear units are connected by inter\u00admolecular N\u2014H\u22efCl hydrogen bonds, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.526\u2005(2) and 3.696\u2005(2)\u2005\u00c5], forming a two-dimensional layered structure parallel to (010).In the centrosymmetric binuclear title complex, [Hg DOI: 10.1107/S1600536811029886/su2294Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dimeric aggregates thus formed are arranged in rows with their terminal NH2 groups forming N\u2014H\u22efO hydrogen bonds with neighbouring aggregates to form a two-dimensional array in the ac plane with an overall T-shaped topology. Layers inter\u00addigitate along the b axis being connected by C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 [centroid\u2013centroid distance = 3.6316\u2005(19)\u2005\u00c5] inter\u00adactions.The constituents of the title co-crystal, C DOI: 10.1107/S1600536810034094/hg2707Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atom in the cation of the title salt, [Zn(C2H3O2)(C18H40N4)]ClO4, is five-coordinated by the four N atoms of the macrocycle and the O atom of the monodentate acetate ligand. The N4O donor set is based on a trigonal bipyramid with two N atoms occupying axial positions [N\u2014Zn\u2014N = 170.89\u2005(16)\u00b0]. The perchlorate anions are associated with the cations via N\u2014H\u22efO hydrogen bonds; intra\u00admolecular N\u2014H\u22efO(acetate) inter\u00adactions are also observed. The neutral aggregates are connected into an helical chain along the b axis via N\u2014H\u22efO(acetate) hydrogen bonds. The perchlorate anion was found to be disordered about a pseudo-threefold axis: the major component of the disorder had a site occupancy factor of 0.692\u2005(11).The Zn N-substituted derivatives and their metal complexes, see: Bembi et al. (C18H40N4)]ClO4 = 0.080 wR(F 2) = 0.183 S = 1.29 4534 reflections320 parameters22 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.63 e \u00c5\u22123 \u0394\u03c1min = \u22120.48 e \u00c5\u22123 \u0394\u03c1 CrystalClear used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811045582/hb6470Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In each, two \u03ba2N:N-chelating 5-(pyridin-2-yl)pyrazine-2-carbonitrile ligands surround the AgI atom, giving an N4O square-pyramidal coordination geometry with one trifluoro\u00adacetate O atom at the apex. The difference between the two lies in the Ag\u2014N bond lengths: in one complex, three normal [range 2.272\u2005(5)\u20132.552\u2005(5)\u2005\u00c5] and one long [2.706\u2005(4)\u2005\u00c5] and in the second, two normal [2.254\u2005(5) and 2.290\u2005(5)\u2005\u00c5] and two long [2.647\u2005(5) and 2.675\u2005(5)\u2005\u00c5] are present. Short inter\u00admolecular F\u22efF contacts [2.586\u2005(4)\u2005\u00c5] and weak \u03c0\u2013\u03c0 stacking inter\u00adactions [minimum ring centroid separation 3.836\u2005(5)\u2005\u00c5] between pyridyl and pyrazinyl rings connect the complex units, forming columns which extend along the b-axis direction.In the asymmetric unit of the title compound, [Ag(C DOI: Click here for additional data file.10.1107/S1600536812040846/zs2235Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CuII ion has a square-pyramidal coordination sphere, the basal plane being occupied by four N atoms in a nearly planar array [mean deviation = 0.048\u2005(6)\u2005\u00c5] with the CuII ion displaced slightly from the plane [0.167\u2005(5)\u2005\u00c5] toward the fifth ligand. The apical position is occupied by a coordinating water mol\u00adecule [Cu\u2014O = 2.319\u2005(4)\u2005\u00c5]. The crystal structure is stabilized by hydrogen-bonding inter\u00adactions between the water mol\u00adecules and carbonyl O atoms. The inversion-related square-pyramidal complex molecules pack base-to-base with long Cu\u22efNpy contact distances of 3.537\u2005(9)\u2005\u00c5, preventing coordination of a sixth ligand.The title compound, [Cu(C II complexes, see: Landee & Turnbull (N3)(H2O)] = 0.063wR(F2) = 0.105S = 1.172258 reflections205 parameters2 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.50 e \u00c5\u22123\u0394\u03c1min = \u22120.69 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, New_Global_Publ_Block. DOI: 10.1107/S1600536813027499/sj5358Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The central Fe2S2 core is in a butterfly conformation and each FeI atom has a pseudo-square-pyramidal coordination by three O atoms and two S atoms. The Fe\u2014Fe distance is 2.471\u2005(2)\u2005\u00c5 and the dihedral angle between the two Fe\u2014S\u2014Fe planes is 78.96\u2005(7)\u00b0. The least-squares plane through the \u2013S(C7H6S)S\u2013 bridge nearly bis\u00adects the mol\u00adecular structure: except for the two Fe(CO)3 units, all atoms are in this plane with an average deviation from the plane of 0.028\u2005(3)\u2005\u00c5. In the crystal, the mol\u00adecules are linked into chains along [001] by C\u2014H\u22ef\u03c0(arene) inter\u00adactions.The title compound, [Fe DOI: Click here for additional data file.10.1107/S1600536813009860/vn2067Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The organic ligand is bidentate, coordinating the ZnII atom via the two N atoms. The benzene and pyridine rings are oriented at a dihedral angle of 11.67\u2005(9)\u00b0. In the crystal, weak C\u2014H\u22efI and C\u2014H\u22efO hydrogen bonds are observed, in addition to \u03c0\u2013\u03c0 stacking inter\u00adactions, with a centroid\u2013centroid distance of 3.72\u2005(5)\u2005\u00c5.In the title complex, [ZnI DOI: 10.1107/S1600536812030486/br2207Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The crystal packing can be described as ClO4 tetra\u00adhedra and CuN4O2 octa\u00adhedra alternating in a zigzag fashion along the c axis. The structure is stabilized by intermolecular N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, as well as \u03c0\u2013\u03c0 interactions [centroid\u2013centroid distance = 3.7179\u2005(15)\u2005\u00c5].In the title compound, [Cu(ClO DOI: 10.1107/S1600536812029868/zj2084Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The ZnII atoms are connected by the bridging ligands into a layer parallel to (101). O\u2014H\u22efO hydrogen bonds link the layers and the dimethyl\u00adformamide solvent mol\u00adecules. \u03c0\u2013\u03c0 inter\u00adactions between the pyridine and benzene rings [centroid\u2013centroid distances = 3.7428\u2005(17) and 3.7619\u2005(17)\u2005\u00c5] and intra\u00adlayer O\u2014H\u22efO hydrogen bonds are also present.In the title compound, {[Zn(C DOI: Click here for additional data file.10.1107/S1600536812043838/hy2597Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The 4,4\u2032-bpy ligand has an inversion center at the mid-point of the central C\u2014C bond. The empty side of the metal ion is capped by two carboxyl\u00adate O atoms from a neighboring mol\u00adecule, with weak Pb\u22efO contacts [Pb\u22efO = 3.069\u2005(2) and 3.071\u2005(3)\u2005\u00c5]. The crystal structure is stabilized by C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the benzene and pyridine rings [centroid\u2013centroid distance = 3.749\u2005(3)\u2005\u00c5].In the title dinuclear complex, [Pb DOI: 10.1107/S1600536811022422/hy2436Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Its structure is isotypic with the CoII analogue. The crystal structure is built up from centrosymmetric dinuclear complex mol\u00adecules and the structure is reinforced by a net of inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds. One water mol\u00adecule is bound to the NiII atom in the octahedral coordination sphere, while the second is part of the inter\u00admolecular hydrogen-bond system.The title compound {sys\u00adtematic name: bis(\u03bc II compound, see: Man et al. 4(H2O)2]\u00b72H2O = 0.046wR(F2) = 0.124S = 1.073522 reflections298 parameters6 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 2.44 e \u00c5\u22123\u0394\u03c1min = \u22120.67 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812022532/zj2074Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, the complex mol\u00adecules and uncoordinated water mol\u00adecules are linked via inter\u00admolecular O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds, forming a three-dimensional supra\u00admolecular network. \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings provide additional stability of the crystal packing [centroid\u2013centroid distance = 3.792\u2005(2)\u2005\u00c5].In the title compound, [Ni(C DOI: 10.1107/S1600536811051063/hy2491Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each Ni2+ cation is coordinated in a cis-mode by two tridentate N,N\u2032,S-chelating Schiff base ligands, creating a distorted octa\u00adhedron . The dihedral angle between the mean coordination planes of the two ligands is 86.76\u2005(7)\u00b0 for one and 89.99\u2005(7)\u00b0 for the second mol\u00adecule. \u03c0\u2013\u03c0 inter\u00adactions between neighbouring pyridine rings with plane-to-plane distances of 3.540\u2005(1) and 3.704\u2005(1)\u2005\u00c5 are observed.The asymmetric unit of the title compound, [Ni(C DOI: 10.1107/S1600536812017333/wm2622Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Cu atom binds to four O atoms, resulting in a four-coordinate square-planar complex. The asymmetric unit contains half of the complex, the other half being completed by inversion symmetry. The Cu\u2014O bond lengths have similar distances, viz. 1.9153\u2005(10)\u2005\u00c5 for the pair opposite (trans) each other and 1.9373\u2005(10)\u2005\u00c5 for the other (trans) pair. The P\u2014O bond lengths are 1.5250\u2005(11)\u2005\u00c5, indicating significant electron delocalization across the O\u2014P\u2014C\u2014P\u2014O atoms in the chelate ring, resulting in a longer P\u2014O bond length when compared to a formal double-bond P=O character (much shorter at approximately 1.47\u2005\u00c5). The two inter\u00adsecting O\u2014Cu\u2014O angles are both linear at 180\u00b0, whilst the remaining L-shaped O\u2014Cu\u2014O bond angles are 88.26\u2005(5) and 91.74\u2005(5)\u00b0. The C\u2014C\u00a0N fragment is slightly distorted from linearity at 177.44\u2005(19)\u00b0 and the C\u00a0N bond length of 1.151\u2005(2)\u2005\u00c5 indicates predominantly triple-bond character.The title complex, [Cu(C For a b al. 2011 and for al. 2008. For bac al. 2008 26H20NO2P2)2] = 0.033 wR(F 2) = 0.086 S = 1.03 5640 reflections286 parametersH-atom parameters constrainedmax = 0.49 e \u00c5\u22123 \u0394\u03c1min = \u22120.32 e \u00c5\u22123 \u0394\u03c1 COSMO I, global. DOI: 10.1107/S1600536811031564/ff2023Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two axial coordination sites are occupied by carbonyl O atoms of neighbouring glycine mol\u00adecules. The Cu\u2014O distances for the axial O atoms [2.648\u2005(2) and 2.837\u2005(2)\u2005\u00c5] are considerably longer than both the Cu\u2014O [1.9475\u2005(17) and 1.9483\u2005(18)\u2005\u00c5] and Cu\u2014N [1.988\u2005(2) and 1.948\u2005(2)\u2005\u00c5] distances in the equatorial plane, which indicates a strong Jahn\u2013Teller effect. In the crystal, the two-dimensional networks are arranged parallel to (001) and are linked via N\u2014H\u22efO hydrogen bonds, forming a three-dimensional arrangement.The title coordination polymer, [Cu(C DOI: 10.1107/S1600536811031503/su2280Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The four O atoms in the equatorial plane around the CoII cation form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two pyridine N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 29.7\u2005(4)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 83.17\u2005(15)\u00b0. Intra\u00admolecular O\u2014H\u22efO hydrogen bonding occurs between the carboxyl\u00adate group and coordinating water mol\u00adecule. In the crystal, inter\u00admolecular N\u2014H\u22efO, O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network.In the title complex, [Co(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.088wR(F2) = 0.237S = 1.233285 reflections202 parametersH atoms treated by a mixture of independent and constrained refinementmax = 1.42 e \u00c5\u22123\u0394\u03c1min = \u22121.61 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536813004984/xu5678Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angle between the benzene rings is 24.8\u2005(2)\u00b0. In the crystal, mol\u00adecules are linked into chains along the b axis by weak C\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions. An inter\u00admolecular Cl\u22efCl [3.4564\u2005(19)\u2005\u00c5] inter\u00adaction is present which is shorter than the sum of the van der Waals radii of Cl atoms (3.50\u2005\u00c5).In the title compound, [Ni(C DOI: 10.1107/S1600536812002085/lh5404Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The bpbd mol\u00adecules bridge the Ag atoms into a chain. Two adjacent chains are further connected by Ag\u22efAg inter\u00adactions [3.1631\u2005(8)\u2005\u00c5], forming a double-chain structure. A \u03c0\u2013\u03c0 inter\u00adaction [centroid\u2013centroid distance = 3.758\u2005(3)\u2005\u00c5] occurs between the double chains. Inter\u00adchain C\u2014H\u22efO hydrogen bonds are observed.In the title compound, [Ag(NO DOI: 10.1107/S1600536810025997/hy2324Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The stereochemistry about each FeCl3O centre is distorted tetra\u00adhedral [Fe\u2014Cl = 2.2176\u2005(5)\u20132.2427\u2005(5)\u2005\u00c5 and Fe\u2014O = 1.7545\u2005(2)\u2005\u00c5]. The Cl atoms are involved in weak anion\u2013cation C\u2014H\u22efCl inter\u00adactions, giving a network structure.In the title compound (C DOI: 10.1107/S1600536810024098/zs2046Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The complex mol\u00adecule and the solvent water mol\u00adecule are located on a twofold rotation axis. In the complex, the MnII ion has a distorted cis-N4O2 octa\u00adhedral coordination geometry defined by four N atoms of the two chelating 2,2\u2032-bipyrimidine ligands and two O atoms of water mol\u00adecules. In the crystal, the complex cations, anions and solvent mol\u00adecules are linked by inter\u00admolecular O\u2014H\u22efN, O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds. The ClO4 \u2212 anion is disordered over two sites with a site-occupancy factor of 0.512\u2005(12) for the major component.The asymmetric unit of the title compound, [Mn(C II complexes, see: Hong et al. 2(H2O)2](ClO4)2\u00b72CH3NO2\u00b7H2O = 0.059 wR(F 2) = 0.199 S = 1.06 3617 reflections208 parametersH-atom parameters constrainedmax = 1.05 e \u00c5\u22123 \u0394\u03c1min = \u22120.69 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536811045612/xu5372Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecule belongs to the symmetry point group Ch2. The CuII ion is located on a twofold rotation axis and the hydroxide and perchlorate ligands are located on a mirror plane. Within the dinuclear mol\u00adecule, the Cu\u22efCu separation is 2.8614\u2005(7)\u2005\u00c5. The crystal structure exhibits O\u2014H\u22efO, C\u2014H\u22efO and \u03c0\u2013\u03c0 [centroid\u2013centroid distance = 3.5374\u2005(13)\u2005\u00c5] inter\u00adactions.In the title binuclear copper(II) complex, [Cu DOI: 10.1107/S1600536813027852/bt6938Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The crystal structure is stablized by weak inter\u00admolecuar C\u2014H\u22ef\u03c0 and N\u2014H\u22ef\u03c0 inter\u00adactions.In the title mol\u00adecule, C DOI: 10.1107/S1600536810029788/lh5094Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The correct Equation should be j = \u03c3 E. An error occurred in Equation 6, j ="} +{"text": "The asymmetric unit contains two independent complex mol\u00adecules as well as two independent benzoic acid solvent mol\u00adecules, one of which is disordered over two positions with almost equal occupancies [0.504\u2005(5) and 0.496\u2005(5)]. The two complex mol\u00adecules have similar configurations with the hexa\u00adcoordinated environment of the PbII atom formed by four carboxyl\u00adate O atoms of two chelate benzoate ligands and two N atoms of the bipy ligand. The Pb\u2014O bonds involving one of the benzoate ligands are almost coplanar with Pb\u2014N bonds to the bipy ligand [dihedral angles of 12.67\u2005(11) and 14.73\u2005(11)\u00b0] ; if the second benzoate ligand is treated as one coordination site, the overall coordination may be represented as a distorted pseudo-square pyramid. Weak inter\u00admolecular Pb\u22efO inter\u00adactions [3.046\u2005(3) and 3.359\u2005(3)\u2005\u00c5] link each of the complex mol\u00adecules into two symmetry-independent centrosymmetric dimers. Hydrogen bonds involving the carboxyl H atoms of solvent benzoic acid mol\u00adecules and metal-coordinated carboxyl\u00adate O atoms link complex mol\u00adecules and benzoic acid solvent mol\u00adecules into insular aggregates.The reaction of lead acetate, benzoic acid and 2,2\u2032-bipyridine (bipy) in aqueous solution yielded the title complex, [Pb(C For the al. 2006; Masaoka al. 2001. 7H5O2)2(C10H8N2)]\u00b7C7H6O2 = 0.035 wR(F 2) = 0.081 S = 1.01 14302 reflections773 parametersH-atom parameters constrainedmax = 1.17 e \u00c5\u22123 \u0394\u03c1min = \u22121.13 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008SHELXTL (Sheldrick, 2008SHELXTL.Data collection: 10.1107/S1600536810046489/ya2132sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536810046489/ya2132Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Both amine H atoms are involved in N\u2014H\u22efS hydrogen bonding, resulting in the formation of layers of inter\u00adlinked mol\u00adecules parallel to the ab plane, which are further held together by weak \u03c0\u2013\u03c0 inter\u00adactions between adjacent complexes, involving one ring of each dipyridyl\u00adamine unit [centroid\u2013centroid distance = 3.777\u2005(4)\u2005\u00c5], forming a three-dimensional assembly.The mononuclear neutral title complex, [Ni(NCS) II, CuII and ZnII complexes with amine ligands, see: Wrzeszcz et al. 2(NCS)2] complexes, see: Wang et al. 2] = 0.066 wR(F 2) = 0.161 S = 1.03 4107 reflections304 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.77 e \u00c5\u22123 \u0394\u03c1min = \u22120.84 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811050197/pk2364Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecular conformation is stabilized by \u03c0\u2013\u03c0 stacking inter\u00adactions [shortest centroid\u2013centroid distance = 3.600\u2005(1)\u2005\u00c5] between substituted benzene rings of different ligands. The crystal packing is characterized by C\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions involving the chloro\u00adform solvent mol\u00adecules.In the title compound, [PdCl DOI: Click here for additional data file.10.1107/S1600536812045801/zq2184Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The effective cone angles for the two phosphane ligands are 160 and 169\u00b0. C\u2014H\u22efCl inter\u00adactions generate infinite long chains along [01-1]. Additional C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking interactions [centroid\u2013centroid distance = 4.2499\u2005(15)\u2005\u00c5 and ring slippage = 2.386\u2005\u00c5] are observed.In the mononuclear title compound, DOI: Click here for additional data file.10.1107/S160053681204696X/ng5305Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, weak C\u2014H\u22efS hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between parallel pyridine rings of adjacent mol\u00adecules [centroid\u2013centroid distance = 3.653\u2005(3)\u2005\u00c5] link the mol\u00adecules into a two-dimensional supra\u00admolecular architecture. The structure contains voids of 124\u2005(9)\u2005\u00c53, which are free of solvent molecules.In the title complex, [Fe(NCS) For rel al. 2008; Min et al. 2008; Phan et al. 2012; Wei et al. 2011.2(C18H18N4)] = 0.069wR(F2) = 0.203S = 1.114456 reflections283 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.42 e \u00c5\u22123\u0394\u03c1min = \u22120.47 e \u00c5\u22123\u0394\u03c1CrystalClear used to solve structure: SHELXTL I, global. DOI: 10.1107/S1600536813034818/xu5762Isup2.hklStructure factors: contains datablock(s) I. DOI: CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The CuII atom occupies a four-coordinate pseudo-tetra\u00adhedral environment bound to one S atom, one imine N atom and one pyridine N atom from the N1,N2-bis\u00ad(pyridin-2-yl)hydrazine-1,2-dicarbo\u00adthio\u00adamidate ligand, and one Cl\u2212 anion. The metal atoms are connected via the bis-tridentate ligand into a binuclear structure. The mol\u00adecule is bow-shaped with the pyridine rings inclined to one another by 51.56\u2005(14)\u00b0. In the crystal, N\u2014H\u22efCl hydrogen bonds lead to the formation of ribbons propagating along [001]. These ribbons are connected via C\u2014H\u22efCl, C\u2014H\u22efS and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.6146\u2005(19)\u2005\u00c5], leading to the formation of a three-dimensional structure.The binuclear title compound, [Cu Cl2] = 0.024wR(F2) = 0.068S = 1.071561 reflections110 parametersH-atom parameters constrainedmax = 0.73 e \u00c5\u22123\u0394\u03c1min = \u22120.34 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global, I. DOI: Click here for additional data file.10.1107/S1600536812048659/su2393Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The two pyridyl planes are inclined with dihedral angles of 59.1\u2005(2) and 61.84\u2005(19)\u00b0 with respect to the PtCl2N2 plane. In the crystal, the complex mol\u00adecules display inter- and intra\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions, with centroid-centroid distances of 3.806\u2005(5)\u20133.845\u2005(5)\u2005\u00c5, which form a one-dimensional column structure along the a axis.In the title complex, DOI: 10.1107/S160053681004393X/is2622Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The environment of Cu2+ ion is a distorted octa\u00adhedron. The axial bond lengths between the CuII ion and the N atoms are considerably longer than the equatorial bond distances between the CuII ion and the N atoms of the ethyl\u00adenediamine ligand as a consequence of the Jahn\u2013Teller effect. The mol\u00adecular conformation is stabilized by intra\u00admolecular N\u2014H\u22efO hydrogen bonds. In the crystal, mol\u00adecules are connected by inter\u00admolecular N\u2014H\u22efO hydrogen bonds into chains running along the a axis.In the crystal structure of the title compound, [Cu(C H)-one 2,2-dioxide], see: Clauss & Jensen 2(C2H8N2)2] = 0.024 wR(F 2) = 0.065 S = 1.09 2023 reflections150 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.23 e \u00c5\u22123 \u0394\u03c1min = \u22120.25 e \u00c5\u22123 \u0394\u03c1 X-AREA I, global. DOI: 10.1107/S1600536811051658/bt5731Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The HgII atoms are four-coordinated in a distorted tetra\u00adhedral geometry by two N atoms from a 2,2\u2032-dimethyl-4,4\u2032-bi-1,3-thia\u00adzole ligand and two Br atoms. In the crystal structure, inter\u00admolecular C\u2014H\u22efBr hydrogen bonds and \u03c0\u2013\u03c0 contacts between the thia\u00adzole rings [centroid\u2013centroid distances = 3.670\u2005(3) and 3.614\u2005(2)\u2005\u00c5] stabilize the structure.The asymmetric unit of the title compound, [HgBr DOI: 10.1107/S1600536810051494/hy2386Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II ion in the title complex, [PtI2(C10H9N3)], is four-coordinated in a distorted square-planar environment defined by the two pyridine N atoms of the chelating di-2-pyridyl\u00adamine (dpa) ligand and by two I\u2212 anions. The dpa ligand is not planar, the dihedral angle between the pyridine rings being 52.8\u2005(3)\u00b0. Pairs of complex mol\u00adecules are assembled through inter\u00admolecular N\u2014H\u22efI hydrogen bonds, forming a dimer-type species. The complexes are stacked in columns along the b axis and display several inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings, with a shortest ring centroid\u2013centroid distance of 3.997\u2005(5)\u2005\u00c5.The Pt II complex [PtCl2(dpa)], see: Li & Liu ] = 0.031wR(F2) = 0.077S = 1.072527 reflections145 parametersH-atom parameters constrainedmax = 1.47 e \u00c5\u22123\u0394\u03c1min = \u22121.31 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812011907/wm2603Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the ligand, the hy\u00addroxy group is involved in an intra\u00admolecular O\u2014H\u22efN hydrogen bond and the two aromatic rings form a dihedral angle of 5.5\u2005(1)\u00b0. In the crystal, weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings [centroid\u2013centroid distance = 3.816\u2005(3)\u2005\u00c5] link the mol\u00adecules into centrosymmetric dimers.In the title compound, [Sn(CH DOI: 10.1107/S1600536811025621/cv5115Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "IV atom in the title complex, [V(C19H17N3O2S)O], is coordinated by two N and two O atoms of the dianionic tetra\u00addentate Schiff base ligand and the terminal oxide O atom. The N2O3 donor set defines a square-pyramidal coordination geometry with the oxide O atom in the apical site. Some buckling in the tetra\u00addentate ligand is indicated by the dihedral angle of 17.92\u2005(19)\u00b0 between the six-membered chelate rings. Supra\u00admolecular chains are formed along the b axis via C\u2014H\u22efO contacts in the crystal. The chains are connected into a layer in the ab plane via C\u2014H\u22ef\u03c0 inter\u00adactions. The atoms comprising the \u2013SCH2\u2014CH=CH2 and methyl substituents were found to be disordered in a 0.916\u2005(2):0.088\u2005(2) ratio. The crystal studied was found to be twinned by nonmerohedry with a 28.1\u2005(4)% minor twin component.The V O] = 0.076wR(F2) = 0.191S = 1.204134 reflections257 parameters3 restraintsH-atom parameters constrainedmax = 0.98 e \u00c5\u22123\u0394\u03c1min = \u22120.94 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812029819/hb6874Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Two dppm ligands, two dpp ligands and two trifluoro\u00adacetate anions bridge four metal atoms, forming a centrosymmetric tetra\u00adnuclear complex. Intra\u00admolecular C\u2014H\u22efO hydrogen bonds and a weak \u03c0\u2013\u03c0 inter\u00adaction [centroid\u2013centroid distance = 3.9804\u2005(13)\u2005\u00c5] are also observed.In the cation of the title compound, [Ag DOI: 10.1107/S1600536811045466/rz2658Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "It is chelated by four N atoms of two different 5,5\u2032-dimethyl-2,2\u2032-bipyridyl (dmbpy) ligands in axial and equatorial positions, and by two O atoms of a carbonate anion completing the equatorial positions. Although the water mol\u00adecules are disordered and their H atoms were not located, there are typical O\u22efO distances between 2.8 and 3.0\u2005\u00c5, indicating O\u2014H\u22efO hydrogen bonding. The crystal packing is consolidated by C\u2014H\u22efO and C\u2014H\u22efBr hydrogen bonds, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions between adjacent pyridine rings of the dmbpy ligands, with centroid\u2013centroid distances of 3.694\u2005(3) and 3.7053\u2005(3)\u2005\u00c5.In the title complex, [Co(CO DOI: 10.1107/S160053681200894X/wm2590Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The organic cation is essentially planar, with a maximum deviation of 0.013\u2005(1)\u2005\u00c5. In the crystal structure, the ions and mol\u00adecules are linked into a pseudo-layered three-dimensional supra\u00admolecular network via O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds. Weak inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions further stabilize the crystal structure [centroid\u2013centroid distance = 3.5231\u2005(4)\u2005\u00c5].In the title compound, (C DOI: 10.1107/S1600536810026693/hb5536Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the mol\u00adecules are packed via \u03c0\u2013\u03c0 stacking inter\u00adactions alternating between imidazole and benzene rings [mean inter\u00adplanar distances = 3.754\u2005(3) and 3.624\u2005(3)\u2005\u00c5]. An inter\u00admolecular N\u2014H\u22efO hydrogen bond links the dimers together. The two-coordinate CuI atom displays an O\u2014Cu\u2014N bond angle of 176.3\u2005(2)\u00b0. The Cu\u22efCu distance within the dimer is 5.100\u2005(2)\u2005\u00c5.The dimeric title complex, [Cu DOI: 10.1107/S1600536810046040/om2365Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One of the water mol\u00adecules is coordinated with the ZnII ion and this mol\u00adecule forms an O\u2014H\u22efO inter\u00adaction with the lattice water mol\u00adecule. The pyridine-2,6-dicarboxyl\u00adate ligand is almost planar (r.m.s. deviation = 0.0242\u2005\u00c5). In the crystal, C\u2014H\u22efO, C\u2014H\u22efN, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds are present.In the title compound, [Zn(C DOI: 10.1107/S1600536811015571/zj2009Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The metal atom is surrounded by the two dimethyl sulfoxide (DMSO) ligands, each coordinating through the O atom, and two anionic saccharinate ligands coordinating through the N atom. The tetra\u00adhedral geometry is slightly distorted as is evident from the N\u2014Zn\u2014N bond angle of 113.85\u2005(6)\u00b0, the O\u2014Zn\u2014O bond angle of 98.92\u2005(6)\u00b0 and O\u2014Zn\u2014N bond angles of 116.96\u2005(6) and 103.93\u2005(6)\u00b0. The Zn\u2014N bond lengths are 1.9742\u2005(15) and 2.0025\u2005(16)\u2005\u00c5. The Zn\u2014O bond lengths are 1.9806\u2005(14)\u2005\u00c5 and 1.9468\u2005(14)\u2005\u00c5. The DMSO ligand coordinates through the lone pair of electrons on the O atom, as can be seen from the Zn\u2014O\u2014S bond angle of 131.30\u2005(8)\u00b0.The title compound, [Zn(C DOI: 10.1107/S1600536811045703/fj2470Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One uncoordinated dmphen mol\u00adecule is situated on a crystallographic twofold axis and the asymmetric unit is completed by one water mol\u00adecule. In the crystal, mol\u00adecules form a one-dimensional framework in the [001] direction through O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds. The crystal packing is further stabilized by \u03c0\u2013\u03c0 stacking inter\u00adactions between the dmphen rings of neighboring mol\u00adecules, with a centroid\u2013centroid separation of 3.5641\u2005(8)\u2005\u00c5 and a partially overlapped arrangement of parallel dmphen rings with a distance of 3.407\u2005(2)\u2005\u00c5.In the title compound, 2[Co(C DOI: 10.1107/S1600536811035148/bh2369Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The complex mol\u00adecules are connected into a one-dimensional structure along [001] via N\u2014H\u22efN hydrogen bonds and further into a three-dimensional structure via N\u2014H\u22efCl hydrogen bonds. \u03c0\u2013\u03c0 inter\u00adactions between the pyrazine and benzene rings and between the benzene rings [centroid\u2013centroid distances = 3.5635\u2005(15) and 3.9128\u2005(16)\u2005\u00c5] are present.In the title complex, [CuCl(C DOI: 10.1107/S1600536812005582/hy2514Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The distorted octa\u00adhedral trans-N2S4 donor set for the Cd2+ ion is defined by two symmetrically S,S\u2032-chelating dithio\u00adcarbamate anions and two pyridine N atoms derived from two monodentate 4-pyridine\u00adaldazine (or 4-{[(pyridin-4-yl\u00admethyl\u00adidene)hydrazinyl\u00adidene}meth\u00adyl]pyridine) mol\u00adecules [dihedral angle between the aromatic rings = 17.33\u2005(8)\u00b0]. In the crystal, mol\u00adecules are connected into a supra\u00admolecular chain via O\u2014H\u22efN hydrogen bonds involving the 4-pyridine\u00adaldazine N atoms not involved in coordination to cadmium. Weak C\u2014H\u22efO and C\u2014H\u22efN links consolidate the packing.The complete mol\u00adecule of the title compound, [Cd(C DOI: 10.1107/S1600536811004508/hb5795Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The NiII atoms are bridged by the 3-amino\u00adpyridine ligands [Ni\u22efN = 6.7048\u2005(3)\u2005\u00c5] and Cl\u2212 atoms [Ni\u22efN = 3.5698\u2005(3)\u2005\u00c5], forming two-dimensional nets. The DMF solvent mol\u00adecule and the non-bridged Cl\u2212 ions participate in N\u2014H\u22efO and N\u2014H\u22efCl hydrogen bonds with the amino groups. The title compound, {[NiCl DOI: 10.1107/S1600536812036215/xu5609Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atom in the title salt, [Cu(C10H24N4)(H2O)2](C6F5CO2)2\u00b72H2O, is chelated by the four N atoms of the 1,4,8,11-tetra\u00adaza\u00adcyclo\u00adtetra\u00addecane (cyclam) ligand and is coordinated by two water mol\u00adecules in a Jahn\u2013Teller-type tetra\u00adgonally distorted octa\u00adhedral geometry. The CuII atom lies on a center of inversion. The cations, anions and uncoordinated water mol\u00adecules are linked by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, forming a layer structure parallel to (001).The Cu DOI: 10.1107/S1600536810025705/bt5287Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Co atom is 0.0187\u2005(8)\u2005\u00c5 out of the mean plane of the four equatorial N atoms. The structure has an O\u22efH\u22efO bridge, which is very common in cobaloxime derivatives, with O\u22efH distances of 1.24\u2005(2) and 1.25\u2005(2)\u2005\u00c5. In the title compound, [Co(C DOI: 10.1107/S1600536811051397/vn2023Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II ion in the title complex, [PdI2(C10H9N3)], is four-coordinated in a distorted square-planar environment defined by the two pyridine N atoms of the chelating di-2-pyridyl\u00adamine (dpa) ligand and two I\u2212 anions. The dpa ligand is not planar, the dihedral angle between the pyridine rings being 51.2\u2005(2)\u00b0. In the crystal, pairs of complex mol\u00adecules are assembled through inter\u00admolecular N\u2014H\u22efI hydrogen bonds into dimeric species. The complexes are stacked in columns along the b axis and display several inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings, with a shortest ring centroid\u2013centroid distance of 3.957\u2005(3)\u2005\u00c5.The Pd II complex [PtI2(dpa)], see: Ha ] = 0.028wR(F2) = 0.065S = 1.013152 reflections145 parametersH-atom parameters constrainedmax = 1.17 e \u00c5\u22123\u0394\u03c1min = \u22120.78 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812033946/tk5137Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The four O atoms in the equatorial plane form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 9.14\u2005(9)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 82.18\u2005(8)\u00b0. In the crystal, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a two-dimensional network lying parallel to (101). \u03c0\u2013\u03c0 stacking between parallel pyridine rings of adjacent mol\u00adecules [centroid\u2013centroid distance = 3.7765\u2005(8)\u2005\u00c5] further stabilizes the crystal structure.In the title complex, [Co(C N,N-di\u00adethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.027wR(F2) = 0.074S = 1.112797 reflections204 parameters52 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.32 e \u00c5\u22123\u0394\u03c1min = \u22120.33 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S160053681301458X/su2606Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atom in the title complex, [Ni(C8H5N5O3)(C2H8N2)(H2O)]\u00b72H2O, is six-coordinated in a distorted octa\u00adhedral geometry by a tridentate 2-amino-7-methyl-4-oxidopteridine-6-carboxyl\u00adate (pterin) ligand, a bidentate ancillary ethane-1,2-diamine (en) ligand and a water mol\u00adecule. The pterin ligand forms two chelate rings. The en and pterin ligands are arranged nearly orthogonally [dihedral angle between the mean plane of the en mol\u00adecule and the pterin ring = 77.1\u2005(1)\u00b0]. N\u2014H\u22efO, O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds link the complex mol\u00adecules and lattice water mol\u00adecules into a three-dimensional network. \u03c0\u2013\u03c0 inter\u00adactions are observed between the pyrazine and pyrimidine rings [centroid\u2013centroid distance = 3.437\u2005(2)\u2005\u00c5].The Ni DOI: Click here for additional data file.10.1107/S160053681300069X/hy2612Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the cations, the ZnII ions have distorted trigonal\u2013bipyramidal environments formed by four N atoms from two 2-chloro-1,10-phenanthroline (cphen) ligands and one N atom from a thio\u00adcyanate ligand. The ZnII atoms in the complex anions also have distorted trigonal\u2013bipyramidal environments, formed by two N atoms from a cphen ligand and three N atoms from three thio\u00adcyanato ligands. The crystal packing exhibits \u03c0\u2013\u03c0 inter\u00adactions between the rings of the cphen ligands [shortest centroid\u2013centroid distance = 3.586\u2005(5)\u2005\u00c5] and short inter\u00admolecular S\u22efCl [3.395\u2005(5)\u2005\u00c5] and S\u22efS [3.440\u2005(4)\u2005\u00c5] contacts.The asymmetric unit of the title compound, [Zn(NCS)(C DOI: 10.1107/S1600536812002280/cv5234Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the complex, the MnII ion is six-coordinated in a distorted octa\u00adhedral environment defined by four N atoms of the two chelating 2,2\u2032-bipyrimidine (bpym) ligands, one I\u2212 anion and one O atom of a water ligand. The dihedral angle between the least-squares planes of the two bpym ligands [maximum deviation = 0.092\u2005(7)\u2005\u00c5] is 79.9\u2005(1)\u00b0. In the crystal, the complex, anion and solvent water mol\u00adecule are linked by inter\u00admolecular O\u2014H\u22efO, O\u2014H\u22efI and O\u2014H\u22efN hydrogen bonds.The asymmetric unit of the title compound, [MnI(C II complexes, see: Hong et al. 2(H2O)]I\u00b7H2O = 0.052 wR(F 2) = 0.126 S = 1.07 5309 reflections262 parametersH-atom parameters constrainedmax = 1.69 e \u00c5\u22123 \u0394\u03c1min = \u22121.98 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 I. DOI: 10.1107/S160053681103875X/bv2192Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two NdIII atoms are linked by two bridging bidentate carboxyl\u00adate groups and two bidentate chelating bridging carboxyl\u00adate groups, with an Nd\u22efNd separation of 4.1259\u2005(4)\u2005\u00c5. Each NdIII atom is nine-coordin\u00adated by five O atoms from the carboxyl\u00adate groups of the zwitterionic azaniumylbenzoate ligands and four from water mol\u00adecules. They adopt a distorted tricapped trigonal\u2013prismatic arrangement. The dihedral angle between the mean planes of the benzene ring and the carboxlate groups are 7.7\u2005(6) and 24.4\u2005(5)\u00b0. The two carboxyl\u00adate groups are almost perpendicular to one another with a dihedral angle of 84.0\u2005(7)\u00b0, while the two benzene rings are inclined to one another by 81.8\u2005(2)\u00b0. The mol\u00adecular packing is stabilized by O\u2014Hwater\u22efCl, O\u2014Hwater\u22efN, N\u2014H\u22efCl, N\u2014H\u22efO, and O\u2014Hwater\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.500\u2005(3)\u2005\u00c5] between symmetry-related benzene rings. All of the Cl\u2212 anions and the uncoordinated water molecules are disordered over two sets of sites with different occupancy ratios.The structure of the title compound, [Nd DOI: 10.1107/S1600536811012700/su2265Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the mol\u00adecules are linked by inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the triazole and benzene rings [centroid\u2013centroid distance = 3.794\u2005(3)\u2005\u00c5] into a band extending in [010]. These bands are further connected by C\u2014H\u22efN hydrogen bonds into a two-dimensional network parallel to (100).In the title complex, [HgI DOI: 10.1107/S160053681302518X/hy2634Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The asymmetric unit is completed by one 4-meth\u00adoxy\u00adbenzoate anion, one nicotinamide (NA) ligand and one coordinated and one uncoordinated water mol\u00adecule. All ligands act in a monodentate mode. The four O atoms in the equatorial plane around the CoII ion form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two pyridine N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the attached benzene ring is 6.47\u2005(7)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 72.80\u2005(4)\u00b0. An O\u2014H\u22efO hydrogen bond links the uncoordinated water mol\u00adecule to one of the carboxyl\u00adate groups. In the crystal structure, inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network.In the mononuclear title compound, [Co(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2]\u00b72H2O = 0.027 wR(F 2) = 0.074 S = 1.07 3776 reflections230 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.55 e \u00c5\u22123 \u0394\u03c1min = \u22120.31 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S1600536810026462/cv2740sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810026462/cv2740Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Three Cl atoms of the [ZnCl4]2\u2212 tetra\u00adhedron act as acceptors in N\u2014H\u22efCl hydrogen bonds. The hydrogen bonds, both of which are bifurcated, lead to the formation of a three-dimensional network. Within the network, inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions with a centroid\u2013centroid distance of 3.5911\u2005(7)\u2005\u00c5 arrange the 4-(dimethyl\u00adamino)\u00adpyridinium cations into anti\u00adparallel dimers.In the title compound, (C DOI: 10.1107/S1600536811005010/rk2262Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ppy ligands are not planar, the dihedral angles between the pyridine and benzene rings being 49.0\u2005(3) and 47.3\u2005(3)\u00b0. In the crystal, the complex mol\u00adecules are stacked in columns along the a axis. In the columns, there are numerous intra- and inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the six-membered rings, the shortest ring centroid\u2013centroid distance being 3.774\u2005(6)\u2005\u00c5.In the title complex, [PtBr II and PdII complexes, cis-[PtCl2(ppy)2] and trans-[PdX2(ppy)2] (X = Cl or I), see: Yoshinari et al. 2] = 0.028wR(F2) = 0.068S = 1.042931 reflections244 parameters2 restraintsH-atom parameters constrainedmax = 1.73 e \u00c5\u22123\u0394\u03c1min = \u22120.93 e \u00c5\u22123\u0394\u03c1Absolute structure: Flack 1983, 856 FriFlack parameter: \u22120.037 (13)SMART used to solve structure: SHELXS97 global. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Two NdIII ions are bridged by four carboxyl\u00adate groups in bi- and tridentate modes, forming a centrosymmetric dinuclear unit, with an Nd\u22efNd distance of 4.0021\u2005(5)\u2005\u00c5, and intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings [centroid\u2013centroid distance = 3.960\u2005(2)\u2005\u00c5]. Inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.820\u2005(2) and 3.804\u2005(2)\u2005\u00c5] and O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds connect the dinuclear mol\u00adecules into a three-dimensional supra\u00admolecular network.In the title compound, [Nd DOI: 10.1107/S1600536811023798/hy2440Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The axial Cu\u2014O distances are significantly longer [2.512\u2005(2)\u2005\u00c5], than the Cu\u2014N [2.022\u2005(2)\u2005\u00c5] and Cu\u2014Cl [2.3232\u2005(4)\u2005\u00c5] distances as a result of Jahn\u2013Teller distortion. Aqua ligands are involved in intra- and inter\u00admolecular hydrogen bonding, and N\u2014H\u22efO inter\u00admolecular hydrogen bonds are formed between the organic ligands. In addition, weak \u03c0\u2013\u03c0 inter\u00adactions are observed between the benzene rings of the ligand [centroid\u2013centroid distance = 3.678\u2005(1)\u2005\u00c5].In the title complex, [CuCl H)-one was reported by Vaillancourt et al. (1998H)-one, see: Turgunov & Englert 2(H2O)2] = 0.032 wR(F 2) = 0.089 S = 1.10 1725 reflections137 parameters3 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.37 e \u00c5\u22123 \u0394\u03c1min = \u22120.46 e \u00c5\u22123 \u0394\u03c1 CrysAlis PRO used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008XP (Bruker, 1998publCIF (Westrip, 2010Data collection: 10.1107/S1600536810048890/nk2075sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810048890/nk2075Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Hf\u2014O bond lengths vary from 2.073\u2005(2) to 2.244\u2005(2)\u2005\u00c5 and the O\u2014Hf\u2014O bite angles vary from 73.49\u2005(9) to 75.60\u2005(9)\u00b0. Weak O\u2014H\u22efO hydrogen-bonding inter\u00adactions are observed between the bridging hy\u00addroxy groups and the dimethylformamide solvent mol\u00adecules. The unit cell contains solvent-accessible voids of 131\u2005\u00c53, but the residual electron density in the difference Fourier map suggests no solvent mol\u00adecule occupies this void.The binuclear molecule of the title compound, [Hf O,O\u2032-and N,O-bidentate ligands with hafnium(IV) and zirconium(IV) to exploit possible separation techniques and for the crystal structures of hafnium(IV) and zirconium(IV) complexes, see: Viljoen et al. 6(OH)2]\u00b72C3H7NO = 0.030 wR(F 2) = 0.072 S = 1.03 8726 reflections475 parameters1 restraintH atoms treated by a mixture of independent and constrained refinementmax = 1.33 e \u00c5\u22123 \u0394\u03c1min = \u22120.97 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SIR92 I, global. DOI: 10.1107/S1600536811049543/pv2484Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "As a result of this difference in the coordination modes, the C\u2014S bond lengths are different, viz. 1.687\u2005(2) and 1.692\u2005(2)\u2005\u00c5 in the bidentate ligand and 1.723\u2005(2)\u2005\u00c5 in the monodentate ligand, whereas the non-coordinating S atom has a C\u2014S distance of 1.649\u2005(2)\u2005\u00c5. The crystal packing is stabilized by C\u2014H\u22efO inter\u00adactions.The title compound, [Pd(C DOI: 10.1107/S1600536811040487/bt5655Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the 2-trifluoro\u00admethyl-1H-benzimidazolium cations link to the [HgCl4]2\u2212 complex anions and lattice water mol\u00adecules via N\u2014H\u22efCl and N\u2014H\u22efO hydrogen bonds, and the lattice water mol\u00adecules further link to the Hg complex anion and the organic cations via O\u2014H\u22efCl and O\u2014H\u22efF hydrogen bonding. One of the trifluoro\u00admethyl groups is disordered over two orientations in a 0.59\u2005(4):0.41\u2005(4) ratio.In the title compound, (C DOI: 10.1107/S1600536812018855/xu5519Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The NiII atom is coordinated by four PzMe3 mol\u00adecules and two thio\u00adcyanate anions to define a trans N4S2 distorted octa\u00adhedral geometry. A number of intra\u00admolecular N\u2014H\u22efN, N\u2014H\u22efS and C\u2014H\u22efN inter\u00adactions contribute to the stability of the complex. The crystal structure is stabilized by inter\u00admolecular N\u2014H\u22efS inter\u00adactions, which link neighbouring mol\u00adecules into chains along the a axis.In the title compound, [Ni(NCS) DOI: 10.1107/S1600536811041419/tk2796Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dpa ligand coordinates the PdII atom in a boat conformation of the resulting chelate ring; the dihedral angle between the pyridine rings is 39.3\u2005(2)\u00b0. The two acetate anions coordinate the PdII atom as monodentate ligands and are located on the same sides of the PdN2O2 unit plane. The carboxyl\u00adate groups of the anionic ligands appear to be delocalized on the basis of the C\u2014O bond lengths. Two complex mol\u00adecules are assembled through inter\u00admolecular N\u2014H\u22efO hydrogen bonds, forming a dimer-type species. Inter\u00admolecular C\u2014H\u22efO hydrogen bonds further stabilize the crystal structure.In the title complex, [Pd(CH II complexes [PdX2(dpa)] (X = Cl or Br), see: Rauterkus et al. 2(C10H9N3)] = 0.049wR(F2) = 0.116S = 0.922925 reflections205 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.99 e \u00c5\u22123\u0394\u03c1min = \u22120.81 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Intra\u00admolecular N\u2014H\u22efO and C\u2014H\u22efO hydrogen-bonding inter\u00adactions stabilize the mol\u00adecular conformation. The dihedral angles formed by the central benzene ring with the outer benzene rings of the terphenyl groups are 47.92\u2005(8), 59.38\u2005(8), 48.24\u2005(8) and 52.37\u2005(8)\u00b0. The dichloro\u00admethane solvent mol\u00adecule inter\u00adacts with the complex mol\u00adecule via a C\u2014H\u22efO hydrogen bond. In the crystal, centrosymmetrically related complex mol\u00adecules are linked into dimers through pairs of C\u2014H\u22efO hydrogen bonds.In the title compound, [Fe(C DOI: 10.1107/S1600536812015553/rz2736Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The NH group of the hydrazone moiety does not form a hydrogen bond, due to a steric crowding. In the crystal, the thio\u00adphene ring takes part in weak \u03c0\u2013\u03c0 stacking inter\u00adactions with the pyridine ring [centroid-to-centroid separation = 3.7553\u2005(19)\u2005\u00c5 and inter\u00adplanar angle = 5.48\u2005(12)\u00b0] and the benzene ring [3.7927\u2005(19)\u2005\u00c5 and 4.58\u2005(12)\u00b0]. Together, these lead to [100] stacks of mol\u00adecules in an alternating head-to-tail arrangement, with two \u03c0\u2013\u03c0 stacking contacts between each adjacent pair.In the title compound, C DOI: 10.1107/S1600536812005673/kp2384Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S1600536812005673/kp2384Isup3.cmlSupplementary material file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II atom in the title compound, [Cu(C16H10ClN2O)2], is located on an inversion center and is tetra\u00adcoordinated by two N and two O atoms from two bidentate 1-[(E)-(2-chloro\u00adphen\u00adyl)diazen\u00adyl]naphthalen-2-olate ligands, forming a square-planar complex. In the crystal, mol\u00adecules are linked via weak C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds, forming chains propagating along [010]. There are also \u03c0\u2013\u03c0 inter\u00adactions present involving adjacent naphthalene rings [centroid\u2013centroid distance = 3.661\u2005(13)\u2005\u00c5].The Cu DOI: 10.1107/S1600536813016681/su2613Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "An intra\u00admolecular O\u2014H\u22efO hydrogen bond occurs. Inter\u00admolecular O\u2014H\u22efO hydrogen bonds link pairs of mol\u00adecules into centrosymmetric dimers. Weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings [shortest centroid\u2013centroid distance = 3.4962\u2005(2)\u2005\u00c5] further stabilize the crystal packing.In the title compound, [Co(C DOI: 10.1107/S1600536811051804/cv5209Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The 1,10-phenanthroline ligand is slightly folded for one complex, the dihedral angle between the pyridine planes being 5.3\u2005(1)\u00b0. In contrast it is nearly planar [0.5\u2005(1)\u00b0] as it complexes with the other HgII atom. The thio\u00adcyanate ligands are virtually linear and the S atom is bonded to HgII with N\u22efS\u2014Hg angles ranging from 99.3\u2005(1) to 103.5\u2005(1)\u00b0. Despite the presence of six aromatic rings in the asymmetric unit, there are no significant inter\u00admolecular \u03c0\u2013\u03c0 contacts between phenanthroline ligands as the centroid\u2013centroid distance of the closest contact between six-membered rings is 4.11\u2005(1) A\u00b0.The asymmetric unit of the title compound, [Hg(SCN) DOI: 10.1107/S1600536812038160/vn2049Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, the anions and cations are linked by N\u2014H\u22efO inter\u00adactions along the b axis and a short N\u2014O\u22ef\u03c0 contact [3.2899\u2005(5)\u2005\u00c5] also occurs.In the title compound, C DOI: 10.1107/S1600536810032629/bx2300Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal packing is stabilized by inter\u00admolecular N\u2014H\u22efO, O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds. There are also \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings of the pydc ligands and between the pydc ligands and the benzene-1,3-diammonium cations, with centroid\u2013centroid distances of 3.4575\u2005(15) and 3.7521\u2005(15)\u2005\u00c5.In the title compound, (C DOI: 10.1107/S1600536811009858/hy2407Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The carboxyl\u00adate groups of the PEB ions are twisted away from the attached benzene rings by 4.0\u2005(6) and 13.3\u2005(5)\u00b0. The two benzene rings of the PEB ions bonded to the same metal ion are oriented at a dihedral angle of 87.4\u2005(3)\u00b0. In the polymeric chain, the NA ligand is linked to one of the carboxyl\u00adate groups via N\u2014H\u22efO hydrogen bonding. In the crystal, adjacent polymeric chains inter\u00adact via N\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds; and the lattice water mol\u00adecule links with the polymeric chains via N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonding. \u03c0\u2013\u03c0 stacking between the benzene and the pyridine rings [centroid\u2013centroid distance = 3.805\u2005(5)\u2005\u00c5] and weak C\u2014H\u22ef\u03c0 inter\u00adactions are also observed in the crystal structure.In the crystal structure of the polymeric title compound, {[Pb(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)]\u00b7H2O = 0.043 wR(F 2) = 0.099 S = 1.06 5076 reflections314 parameters5 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 1.11 e \u00c5\u22123 \u0394\u03c1min = \u22120.96 e \u00c5\u22123 \u0394\u03c1 CrystalClear used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811026535/xu5254Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two Bi atoms and two of the C atoms directly bonded to bis\u00admuth are quasi-planar with the carbonate group. The remaining two ligands are in a trans arrangement relative to the quasi-planar (CBi)2CO3 system. The metal atom is strongly coordinated by the N atom of one pendant arm [Bi\u2014N = 2.739\u2005(6)\u2005\u00c5], almost trans to the O atom, while the N atom of the other pendant arm exhibits a weaker intra\u00admolecular inter\u00adaction [Bi\u22efN = 3.659\u2005(7)\u2005\u00c5] almost trans to a C atom. If both these intra\u00admolecular N\u2192Bi inter\u00adactions per metal atom are considered, the overall coordination geometry at bis\u00admuth becomes distorted square-pyramidal [2BiO cores] and the compound can be described as a hypervalent 12-Bi-5 species. Additional quite short intra\u00admolecular Bi\u22efO inter\u00adactions are also present [3.796\u2005(8)\u20134.020\u2005(9)\u2005\u00c5]. Inter\u00admolecular associations through weak \u03b76\u22efBi inter\u00adactions [Bi\u22efcentroid of benzene ring = 3.659\u2005(1)\u2005\u00c5] lead to a ribbon-like supra\u00admolecular association.The mol\u00adecular structure of the title compound, [Bi DOI: 10.1107/S160053681005453X/rk2252Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the complex cation, the FeII atom is coordinated by six N atoms from three chelating dimephen ligands at an average Fe\u2014N distance of 1.963\u2005(4)\u2005\u00c5 giving a distorted octa\u00adhedral geometry. The crystal structure is stabilized by weak C\u2014H\u22efN hydrogen bonds and C\u00a0N\u22ef\u03c0 inter\u00adactions between planar [maximum deviations of 0.024\u2005(3) and 0.015\u2005(3)\u2005\u00c5] tcm anions and pyridine rings of dimephen .The title compound, [Fe(C DOI: Click here for additional data file.10.1107/S1600536812046880/bx2428Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The deprotonated ligand intra\u00admolecularly hydrogen bonds to the thia\u00adzole ring N atom, while the deprotonated ligand forms an inter\u00admolecular hydrogen bond to the thiol\u00adate S atom. The deprotonation of the tridentate ligand and its coordination to Hg via the S atom strikingly affects the C\u2014S bond lengths. In the free ligand, the C\u2014S bond distance is 1.685\u2005(7)\u2005\u00c5, whereas it is 1.749\u2005(7)\u2005\u00c5 in the deprotonated ligand. Similarly, the Hg\u2014S bond distance is slightly longer to the neutral ligand [2.6682\u2005(18)\u2005\u00c5] than to the deprotonated ligand [2.5202\u2005(19)\u2005\u00c5]. The Hg\u2014I distance is 2.7505\u2005(8)\u2005\u00c5.In the title compound, [Hg(C I(C12H12N4S2)] = 0.042 wR(F 2) = 0.085 S = 1.02 5837 reflections346 parametersH-atom parameters constrainedmax = 1.36 e \u00c5\u22123 \u0394\u03c1min = \u22121.27 e \u00c5\u22123 \u0394\u03c1 COLLECT used to solve structure: SIR97 (Altomare et al., 1999SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997SHELXL97.Data collection: 10.1107/S160053681101974X/vm2089sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S160053681101974X/vm2089Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The isobutyl group is disordered over two sets of sites in a 0.899\u2005(9):0.101\u2005(9) ratio. In the crystal, weak aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions involving the imidazole and thia\u00addiazole rings with a centroid\u2013centroid distance of 3.8067\u2005(7)\u2005\u00c5 occur.In the title compound, C DOI: 10.1107/S1600536810053201/hb5776Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The four O atoms in the equatorial plane around the ZnII cation form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 24.13\u2005(8)\u00b0, while the pyridine ring and the benzene ring are oriented at a dihedral angle of 88.52\u2005(4)\u00b0. The coordinating water mol\u00adecule links with the carboxyl\u00adate group via an O\u2014H\u22efO hydrogen bond. In the crystal, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, and a weak C\u2014H\u22ef\u03c0 inter\u00adaction link the mol\u00adecules into a two-dimensional network parallel to (010). These networks are linked via C\u2014H\u22efO and \u03c0\u2013\u03c0 inter\u00adactions between inversion-related benzene rings [centroid\u2013centroid distance = 3.8483\u2005(7)\u2005\u00c5], forming a three-dimensional supra\u00admolecular structure.In the title complex, [Zn(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.022wR(F2) = 0.060S = 1.063475 reflections216 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.43 e \u00c5\u22123\u0394\u03c1min = \u22120.31 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S160053681203320X/su2483Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The two phen ligands exhibit nearly perfect coplanarity (r.m.s. deviations = 0.027 and 0.031\u2005\u00c5), making a dihedral angle of 85.7\u2005(1)\u00b0. The mean inter\u00adplanar distances of 3.36\u2005(2) and 3.41\u2005(3)\u2005\u00c5 between adjacent phen ligands indicate \u03c0\u2013\u03c0 stacking inter\u00adactions. The uncoordinated water mol\u00adecules are partly occupied. One carboxyl\u00adate O atom and two Br atoms are each disordered over two sites, with occupancy factors of 0.60 and 0.40. In the crystal structure, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions link the complex cations, uncoordinated 4-bromo\u00adbenzoate anions and water mol\u00adecules into a three-dimensional supra\u00admolecular network. An intra\u00admolecular O\u2014H\u22ef\u00b7O hydrogen bond is observed in the cation.In the title compound, [Zn(C DOI: 10.1107/S160053681003864X/hy2357Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The N atoms occupy one axial and one equatorial site and the terminal chloride ion occupies an equatorial site. The dihedral angle between the pyridine and pyrazole rings is 12.8\u2005(2)\u00b0. In the crystal, aromatic \u03c0\u2013\u03c0 stacking [centroid\u2013centroid separations = 3.812\u2005(3) and 3.848\u2005(3)\u2005\u00c5] and C\u2014H\u22efCl and C\u2014H\u22ef\u03c0 inter\u00adactions help to establish the packing.In the centrosymmetric binuclear title compound, [Zn DOI: 10.1107/S1600536810026127/hb5500Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Adjacent chains are linked by Owater\u2014H\u22efO hydrogen-bonding inter\u00adactions. The Cu(OH2)3O4 cluster displays a penta\u00adgonal bipyrimadal geometry with two weak coordinations [Cu\u2014Ofuran = 2.790\u2005(2)\u2005\u00c5) and Cu\u2014Ocarboxyl\u00adate = 2.684\u2005(2)\u2005\u00c5] and two water mol\u00adecules located in axial positions. In the title compound, [Cu(C DOI: 10.1107/S1600536812010161/zj2064Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Ca\u2014O bond lengths are in the range 2.354\u2005(3)\u20132.453\u2005(2)\u2005\u00c5, while the Ca\u2014N bond lengths are in the range 2.523\u2005(2)\u20132.548\u2005(2)\u2005\u00c5. An intra\u00admolecular O\u2014H\u22efO hydrogen bond between the carb\u00adoxy and carboxyl\u00adate groups stabilizes the mol\u00adecular configuration. A three-dimensional network of N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds help to stabilize the crystal packing.In the title compound, [Ca(C DOI: 10.1107/S1600536812035544/ff2078Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "One of the nearly planar quinoline ligands [maximum deviations = 0.042\u2005(6) and 0.018\u2005(7)\u2005\u00c5] is almost perpendicular to the PtCl2N2 unit [maximum deviation = 0.024\u2005(3)\u2005\u00c5], making a dihedral angle of 89.6\u2005(1)\u00b0, whereas the other is slightly inclined to the central plane with a dihedral angle of 74.1\u2005(1)\u00b0. The dihedral angle between the quinoline ligands is 88.3\u2005(2)\u00b0. In the crystal, each solvent mol\u00adecule is linked to the metal complex by weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds.In the title compound, [PtCl DOI: 10.1107/S1600536812012469/xu5489Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The geometry around the CuII ion is highly distorted with the range of Cl\u2014Cu\u2014Cl angles being 94.94\u2005(1)\u2013141.03\u2005(1)\u00b0. The crystal structure is stabilized by N\u2014H\u22efCl and C\u2014H\u22efCl hydrogen bonds. In the three-dimensional network, cations and anions pack in the lattice so as to generate chains of [CuCl4]2\u2212 anions separated by two orientations of cation layers, which are inter\u00adlocked through \u03c0\u2013\u03c0 stacking contacts between pairs of pyridine rings, with centroid\u2013centroid distances of 3.7874\u2005(7)\u2005\u00c5.The asymmetric unit of the title salt, (C DOI: 10.1107/S1600536813028006/bq2389Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The crystal structure is stabilized by inter\u00admolecular N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions.In the title complex, [Ag(C DOI: 10.1107/S1600536811017776/jh2288Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The negatively charged N atom of the group binds to the terminal SnIV atom at a shorter distance [Sn\u2014N = 2.236\u2005(2)\u2005\u00c5] compared with the neutral N atom that binds to the central SnIV atom [Sn\u2190 N = 2.805\u2005(2)\u2005\u00c5]. The terminal SnIV atom is five-coordinate in a cis-C2SnNO2 trigonal\u2013bipyramidal geometry [C\u2014Sn\u2014C = 136.4\u2005(1)\u00b0], whereas the central SnIV atom is six-coordinate in a C2SnNO3 skew-trazepoidal bipyramidal geometry [C\u2014Sn\u2014C = 145.4\u2005(1)\u00b0]. The C atoms of the isopropoxy group are disordered over two positions in a 0.591\u2005(7):0.409\u2005(7) ratio.The tetra\u00adnuclear title compound, [Sn DOI: Click here for additional data file.10.1107/S160053681204771X/bt6864Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The supra\u00admolecular structure is based on inter\u00adactions involving the meth\u00adoxy groups , \u03c0\u2013\u03c0 stacking of the electron-rich meth\u00adoxy-substituted rings [centroid\u2013centroid distances of 3.6454\u2005(9)\u20133.738\u2005(1)\u2005\u00c5] and C\u2014H\u22ef\u03c0 contacts .The title compound, C DOI: 10.1107/S1600536811012888/vm2079Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II\u2013LuIII salen-type complex, [CuLu(C22H24N2O4)(NO3)3], with the ligand 6,6\u2032-dimeth\u00adoxy-2,2\u2032-diphenolate, the irregular nine-coordinate LuIII coordination sphere comprises four O atoms from the ligand and five O atoms from three nitrate groups, two bidentate and one monodentate [Lu\u2014O = 2.230\u2005(3)\u20132.621\u2005(4)\u2005\u00c5]. The slightly distorted square-planar four-coordinate CuII atom comprises two imine N atoms [Cu\u2014N = 1.903\u2005(4) and 1.912\u2005(4)\u2005\u00c5] and two phenolate O atoms from the ligand mol\u00adecule [Cu\u2014O = 1.897\u2005(3) and 1.906\u2005(3)\u2005\u00c5]. All atoms of the cyclo\u00adhexane ring of the ligand mol\u00adecule are disordered over two sets of sites with equal occupancy.In the title dinuclear Cu DOI: 10.1107/S1600536810048245/zs2074Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "This disorder is correlated with the disorder of one of the H atoms of the water mol\u00adecule. The carboxyl group is twisted relative to the attached benzene ring by 75.1\u2005(4)\u00b0. The intra\u00admolecular I\u22efO distance is 2.112\u2005(6)\u2005\u00c5. Mol\u00adecules are linked via O\u2014H\u22efO hydrogen bonding, C\u2014I\u22efO halogen bonding, with I\u22efO distances in the range 3.156\u2005(5)\u20133.274\u2005(6)\u2005\u00c5, and dipolar C=O\u22efC=O inter\u00adactions between the carboxyl and carboxyl\u00adate groups, with an O\u22efC distance of 2.944\u2005(10)\u2005\u00c5.In the title compound, C DOI: 10.1107/S1600536812005351/gk2447Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S1600536812005351/gk2447Isup3.cmlSupplementary material file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The oxide O atoms adopt a cis conformation: one is trans to the methanol O atom and the other is trans to the ligand N atom. The dihedral angle between the two benzene rings in the hydrazone ligand is 4.0\u2005(3)\u00b0. In the crystal, mol\u00adecules are linked by O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds.In the title dioxidomolybdenum(VI) complex, [Mo(C O2(CH4O)] = 0.044wR(F2) = 0.093S = 1.034300 reflections269 parameters1 restraintH atoms treated by a mixture of independent and constrained refinementmax = 0.62 e \u00c5\u22123\u0394\u03c1min = \u22120.69 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812008549/hb6656Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angle between the pyridine rings is 1.92\u2005(11)\u00b0. In the crystal, \u03c0\u2013\u03c0 stacking between parallel pyridine rings of adjacent complex mol\u00adecules is observed, the centroid\u2013centroid distance being 3.6788\u2005(19)\u2005\u00c5. Weak C\u2014H\u22efO hydrogen bonds further link the mol\u00adecules into a three-dimensional supra\u00admolecular architecture.In the title complex, [Cu(NO DOI: 10.1107/S1600536813028201/xu5745Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Li+ ion is N,N\u2032-chelated by the N-silylated amido ligand, with Li\u2014N = 2.015\u2005(5) and 2.074\u2005(5)\u2005\u00c5. The two amido ligands are arranged cis to each other. The mol\u00adecule exhibits a twofold rotational symmetry operation along the Li\u2013Na axis. The Na+ ion is coordinated by two N atoms from the tetra\u00admethyl\u00adethylenediamine ligand [Na\u2014N = 2.553\u2005(4)\u2005\u00c5] and shares two amido N atoms from the N-silylated amido ligands with the Li+ ion. Although the crystal structure contains voids with an approximate volume of 50\u2005\u00c53 there is no inclusion of solvent mol\u00adecules.In the heterometallic title bulky amido complex, [LiNa(C N-silylated quinolyl amido ligands, see: Engelhardt et al. 2(C6H16N2)] = 0.062wR(F2) = 0.195S = 0.974001 reflections209 parameters1 restraintH-atom parameters constrainedmax = 0.32 e \u00c5\u22123\u0394\u03c1min = \u22120.26 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S160053681204576X/rk2385Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The complex mol\u00adecules are connected via water O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds and weak \u03c0\u2013\u03c0 stacking inter\u00adactions between the benzene rings [minimum ring centroid separation = 3.750\u2005(6)\u2005\u00c5] into a three-dimensional polymeric structure. The imidazolyl group of the ligand is partially disordered over two sets of sites with refined occupancies of 0.531\u2005(7):0.469\u2005(7).In the title compound, [Mn(C DOI: 10.1107/S1600536812010446/zs2182Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Inter\u00admolecular C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridyl rings [centroid\u2013centroid distance = 3.7337\u2005(18)\u2005\u00c5] are present in the crystal structure.In the title compound, [CdCl DOI: 10.1107/S1600536810029399/hy2335Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The metal atom is further chelated by a carboxyl\u00adate group and is covalently bonded to a monodentate carboxyl\u00adate group and to a monodentate sulfonate group in a distorted square-anti\u00adprismatic geometry. The coordinating and the solvent water mol\u00adecules are hydrogen bonded to the network. In the crystal, one solvent water mol\u00adecule is disordered over two positions [major component = 59\u2005(3)%].The 4-sulfophthalate trianion in the polymeric title complex, {[Dy(C III complex, see: Zhang et al. (C12H8N2)(H2O)2]\u00b72H2O = 0.037wR(F2) = 0.093S = 1.204022 reflections353 parameters33 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.97 e \u00c5\u22123\u0394\u03c1min = \u22121.85 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 global, I. DOI: 10.1107/S1600536812003613/xu5457Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The benzene rings of the anions are involved in \u03c0\u2013\u03c0 stacking. The centroid\u2013centroid distance between parallel benzene rings of adjacent mol\u00adecules is 3.9017\u2005(17)\u2005\u00c5, and the centroid\u2013centroid distance between benzene and pyridine rings of adjacent mol\u00adecules is 3.584\u2005(2)\u2005\u00c5. Intra\u00admolecular O\u2014H\u22efO hydrogen bonding is present.In the title compound, [Zn(C DOI: 10.1107/S1600536811020435/ng5174Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II ion in the title centrosymmetric dinuclear complex, [Hg2Cl4(C13H12N2)2]\u00b7[HgCl2], adopts a distorted square-pyramidal geometry, being coordinated by the bis-chelating N-heterocyclic ligand, two bridging Cl atoms and one terminal Cl atom. One of the bridging Hg\u2014Cl bonds [2.8428\u2005(11)\u2005\u00c5] is significantly longer than the other [2.5327\u2005(10)\u2005\u00c5]. In the crystal, there are weak \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.630\u2005(3)\u2005\u00c5] between the aromatic rings of the discrete units. The HgCl2 adduct molecule is located on an inversion centre and has an Hg\u2014Cl bond length of 2.2875\u2005(11)\u2005\u00c5.The Hg DOI: 10.1107/S1600536810050725/jh2236Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, mol\u00adecules are linked into sheets parallel to the bc plane by C\u2014H\u22efCl and C\u2014H\u22efO hydrogen bonds and weak \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.669\u2005(1)\u2005\u00c5].In the title compound, [ZnCl DOI: 10.1107/S160053681004119X/ci5172Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, mol\u00adecules are connected by C\u2014H\u22efBr and C\u2014H\u22efO hydrogen bonds, and \u03c0\u2013\u03c0 inter\u00adactions between the oxazole and benzene rings [centroid\u2013centroid distance = 3.7344\u2005(19)\u2005\u00c5], resulting in a three-dimensional supra\u00admolecular structure.In the title compound, [PdBr(C DOI: Click here for additional data file.10.1107/S1600536812049197/hy2606Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Four water O atoms in the equatorial plane around the CoII ion [Co\u2014O = 2.0593\u2005(16) and 2.1118\u2005(16)\u2005\u00c5] form a slightly distorted square-planar arrangement, and the distorted octahedral geometry is completed by the two N atoms [Co\u2014N = 2.1306\u2005(18)\u2005\u00c5] from two isonicotinamide ligands. In the anion, the carboxyl\u00adate group is twisted from the attached benzene ring at 8.84\u2005(17)\u00b0. In the crystal, a three-dimensional hydrogen-bonding network, formed by classical O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, consolidates the crystal packing, which exhibits \u03c0\u2013\u03c0 inter\u00adactions between the benzene and pyridine rings, with centroid\u2013centroid distances of 3.458\u2005(1) and 3.606\u2005(1)\u2005\u00c5, respectively.The asymmetric unit of the title compound, [Co(C DOI: 10.1107/S1600536812003911/cv5239Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angle between the aromatic ring and the pyridyl ring is 71.7\u2005(1)\u00b0. In the crystal, the mol\u00adecules are stacked in columns along the c axis and several inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions are present between the six-membered rings, with a shortest centroid\u2013centroid distance of 3.707\u2005(2)\u2005\u00c5.The title compound, C DOI: 10.1107/S1600536811003011/ng5109Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, adjacent mol\u00adecules are connected by \u03c0\u2013\u03c0 contacts between the thia\u00adzole rings [centroid\u2013centroid distance = 3.591\u2005(3)\u2005\u00c5].In the title compound, [HgI DOI: 10.1107/S1600536810029302/hy2334Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the components are linked by O\u2014H\u22efO, C\u2014H\u22efO and aromatic \u03c0\u2013\u03c0 stacking [shortest centroid\u2013centroid separation = 3.659\u2005(5)\u2005\u00c5] inter\u00adactions.In the title compound, [Co(C DOI: 10.1107/S1600536810022750/hb5497Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The 5-hy\u00addroxy\u00adnicotinate ligand is protonated at the N atom and deprotonated at the hy\u00addroxy group. The HoIII atoms are bridged by the carboxyl\u00adate and phenolate O atoms, forming a three-dimensional framework. N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, as well as \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings [centroid\u2013centroid distance = 3.794\u2005(2)\u2005\u00c5], are observed.In the title compound, {[Ho DOI: 10.1107/S1600536812032916/hy2567Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "One of the Cp ligands is additionally substituted by a dimethyl\u00adamino\u00admethyl group causing the chirality of the mol\u00adecule. Surprisingly, although the synthetic procedure yielded the title compound as a racemic mixture, the reported crystal is enanti\u00adomerically pure with the R absolute configuration. The dimethyl\u00adamino group is exo with respect to the Cp ring. Both diphenyl\u00adthio\u00adphosphine groups are trans with respect to the centroid\u2013Fe\u2013centroid direction. Weak intra\u00admolecular C\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions between symmetry-related mol\u00adecules are observed. The contribution of the disordered solvent was removed from the refinement using SQUEEZE in PLATON . In the title compound, [Fe(CSpek 2009. Acta Cr For the al. 2010; Debono al. 2010; Diab et al. 2008; Le Roux al. 2007; Malaceal. 2006a,b \u25b6; Ro20H21NPS)(C17H14PS)]\u00b7CH2Cl2 = 0.042wR(F2) = 0.102S = 1.097854 reflections393 parametersH-atom parameters constrainedmax = 0.63 e \u00c5\u22123\u0394\u03c1min = \u22120.36 e \u00c5\u22123\u0394\u03c1Absolute structure: Flack 1983, 3441 FrFlack parameter: 0.043 (16)APEX2 used to solve structure: SIR97 I, global. DOI: 10.1107/S1600536812022301/im2377Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The resulting N2O3 donor set defines a distorted square-pyramidal geometry with the coordinated water mol\u00adecule in the apical position. In the crystal, the presence of O\u2014Hw\u22efOc (w = water and c = carbon\u00adyl) hydrogen bonding leads to the formation of a supra\u00admolecular chain propagating along the c axis, which associates into a double chain via C\u2014H\u22ef O and \u03c0\u2013\u03c0 contacts between pyridyl rings [centroid\u2013centroid distance = 3.527\u2005(3)\u2005\u00c5]. The solvent mol\u00adecules, which are disordered over two orientations in a 0.678\u2005(11):0.322\u2005(11) ratio, occupy voids defined by the complex mol\u00adecules and are held in place via C\u2014H\u22efO inter\u00adactions.The title complex, [Cu(C DOI: 10.1107/S1600536810030436/hb5590Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "IV atom in the title compound, [Sn(C6H5)2Cl2(C13H12OS)], displays a distorted C2Cl2O trigonal\u2013bipyramidal coordination environment, with a mean Sn\u2014C distance of 2.121\u2005(9)\u2005\u00c5 and with Sn\u2014O = 2.331\u2005(2)\u2005\u00c5. The SnIV atom is displaced by 0.169\u2005(2)\u2005\u00c5 from the equatorial C2Cl plane towards the direction of the second axially bonded Cl atom.The Sn DOI: 10.1107/S1600536811014474/wm2481Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII atom is tetra\u00adcoordinated, displaying a slightly distorted square-planar geometry. The main deviation from the ideal geometry is seen in the differences in the Cu\u2014O [1.8833\u2005(10)\u2005\u00c5] and Cu\u2014N [1.9405\u2005(13)\u2005\u00c5] bond lengths, while angular deviations are less than 3\u00b0. Intra\u00admolecular O\u2014H\u22efO and inter\u00admolecular C2sp\u2014H\u22efO hydrogen bonds form S(5) and R22(8) ring motifs, respectively. The latter inter\u00adaction results in chains of mol\u00adecules along [100].In the title compound, [Cu(C DOI: 10.1107/S1600536812032187/lr2071Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Three RuCp+ (Cp is cyclo\u00adpenta\u00addien\u00adyl) groups are bonded to the three aromatic rings of the ligand. Surprisingly, the pyramidalized N atom of the heterocycle (\u03a3 C\u2014N\u2014C = 329.0\u00b0) points towards the anthracenyl group, so losing its coordinative ability. There is an inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction involving an acetone mol\u00adecule and the adjacent benzyl ring of the ligand. In the crystal, mol\u00adecules are linked via a number of C\u2014H\u22efO and C\u2014H\u22efF inter\u00adactions and a C\u2014H\u22ef\u03c0 inter\u00adaction, leading to the formation of a three-dimensional supra\u00admolecular structure. One of the Cp groups is disordered over two positions, with refined occupancies of 0.695\u2005(14):0.305\u2005(14). Two of the three hexa\u00adfluoro\u00adphospate anions are disordered, with refined occupancies of 0.630\u2005(6):0.370\u2005(6) and 0.771\u2005(8):0.229\u2005(8). One of the two solvent acetone mol\u00adecules is also disordered, with refined occupancies of 0.82\u2005(2):0.18\u2005(2).In the title compound, [Ru DOI: 10.1107/S1600536812040652/su2499Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CdII ions are connected by two bridging benzoate anions and each ion is seven-coordinated by five O atoms from three benzoate ligands and by two N atoms from 1,10-phenanthroline. The benzoate ligands adopt two different coordination modes, acting as bidentate and bridging tridentate ligands. The discrete neutral mol\u00adecules further extend their structure into a three-dimensional supra\u00admolecular framework by inter\u00admolecular \u03c0\u2013\u03c0 [inter\u00adplanar distances of 3.392\u2005(4)\u2005\u00c5] and C\u2014H\u22ef\u03c0 stacking inter\u00adactions [H\u2013mean plane = 2.567\u2005(4) and 2.781\u2005(4)\u2005\u00c5].The dinuclear title compound, [Cd DOI: 10.1107/S1600536811022185/zq2105Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ZnII ion is coordinated by two N atoms from one mol\u00adecule of the aromatic base and four O atoms from two bidentate, symmetry-related acetate anions, which coordinate asym\u00admetrically [Zn\u2014O distances of 2.058\u2005(2) and 2.362\u2005(3)\u2005\u00c5], while the two Zn\u2014N bond distances are equal as imposed by symmetry [2.079\u2005(2)\u2005\u00c5]. The crystal structure is supported by a number of weak C\u2014H\u22efO inter\u00adactions and C\u2014H\u22ef\u03c0 contacts, with no \u03c0\u2013\u03c0 inter\u00adactions present, mainly hindered by the substituent methyl groups and the relative mol\u00adecular orientation. The result is a three-dimensional structure in which each mol\u00adecule is linked to eight different neighbors.The mol\u00adecular structure of the title compound, [Zn(CH DOI: Click here for additional data file.10.1107/S1600536812042699/br2212Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "It should be noted, that a solvent-masking procedure as implemented in OLEX2 [Dolomanov et al. (2009). J. Appl. Cryst.42, 339\u2013341II ion is five-coordinated by two phenanthroline ligands and one chloride ion in a distorted trigonal\u2013bipyramidal geometry. The dihedral angle between the phen ligands is 65.21\u2005(5)\u00b0. The MnII ion is six-coordinated by one Cl atom, two N atoms from a phen ligand, as well one N atom and two O atoms from pydc in a distorted octa\u00adhedral coordination geometry, with cis angles ranging from 72.00\u2005(8) to 122.07\u2005(8)\u00b0 and trans angles ranging from 143.98\u2005(8) to 163.15\u2005(6)\u00b0. In the crystal, C\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds, cation\u2013anion \u03c0\u2013\u03c0 inter\u00adactions between the phen ring systems with centroid\u2013centroid distances in the range 3.881\u2005(34)\u20134.123\u2005(36)\u2005\u00c5, as well as cation\u2013cation, anion\u2013anion \u03c0\u2013\u03c0 inter\u00adactions between the phen rings with centroid\u2013centroid distances in the range 3.763\u2005(4)\u20133.99\u2005(5)\u2005\u00c5 and pydc rings with centroid\u2013centroid distances 3.52\u2005(5)\u2005\u00c5 link the various components. The title complex, [CuCl(C DOI: 10.1107/S1600536814006369/br2237Isup2.hklStructure factors: contains datablock(s) I. DOI: 993053CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dihedral angles between the mean planes of the central phenolate rings and the peripheral phenyl rings are 46.62\u2005(10)/87.06\u2005(9), 60.44\u2005(8)/23.13\u2005(8) and 46.49\u2005(6)/65.29\u2005(6)\u00b0. The crystal packing is stabilized by weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions. Mol\u00adecules are further linked by two \u03c0\u2013\u03c0 [centroid\u2013centroid distances = 3.8612\u2005(14) and 3.9479\u2005(14)\u2005\u00c5] and four C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional network.In the title compound, [Co(C DOI: 10.1107/S1600536814001664/hg5379Isup2.hklStructure factors: contains datablock(s) I. DOI: CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The slightly distorted square-planar coordination sphere of the CuII atom comprises two phenolate O atoms and two oxime N atoms from two bidentate\u2013chelate 2-[1-(eth\u00adoxy\u00adimino)\u00adeth\u00adyl]-1-naphtho\u00adlate O-ethyl oxime (L \u2212) ligands [Cu\u2014O = 1.8919\u2005(17)\u2005\u00c5 and Cu\u2014N = 1.988\u2005(2)\u2005\u00c5]. The two naphthalene ring systems in the mol\u00adecule are parallel, with a perpendicular inter\u00adplanar spacing of 1.473\u2005(2)\u2005\u00c5, while each complex unit forms links to four other mol\u00adecules via inter\u00admolecular methyl C\u2014H\u22ef\u03c0 inter\u00adactions, giving an infinite cross-linked layered supra\u00admolecular structureIn the title complex, [Cu(C DOI: 10.1107/S1600536810047574/zs2078Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ligands are mutually transoid with respect to the metal atom. Weak inter\u00admolecular C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions, with centroid\u2013centroid distances of 3.8452\u2005(14) and 3.9932\u2005(14)\u2005\u00c5, are found in the crystal packing.In the title complex, [ZnCl DOI: 10.1107/S1600536811022884/fj2423Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In each mol\u00adecule, the geometry around the Pd atom is distorted square-planar, with the Pd atom displaced by 0.0549\u2005(12) and 0.0734\u2005(13)\u2005\u00c5 from the least-squares plane of the I\u2014P\u2014P\u2014C atoms. The PPh3 ligands are in trans positions, with P\u2014Pd\u2014P angles of 173.12\u2005(4) and 170.29\u2005(4)\u00b0, while the pyrazinyl ligands and I atoms, also trans to each other, form C\u2014Pd\u2014I angles of 179.38\u2005(12) and 178.44\u2005(12)\u00b0. In the crystal, C\u2014H\u22ef\u03c0 inter\u00adactions occur, resulting in a three-dimensional supramolecular architecture.There are two independent mol\u00adecules with similar configurations in the asymmetric unit of the title complex, [Pd(C I(C18H15P)2] = 0.048wR(F2) = 0.101S = 1.0415924 reflections829 parametersH-atom parameters constrainedmax = 1.35 e \u00c5\u22123\u0394\u03c1min = \u22120.55 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536812043589/bg2480Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The pyrimidine\u00adthiol\u00adate anion acts both as a bridging and a chelating ligand. The AgI ions are linked via two \u03bc2-S donor atoms, which generate a strictly planar Ag2S2 core with an Ag\u22efAg separation of 2.9569\u2005(4)\u2005\u00c5. The AgI ion presents a distorted tetra\u00adhedral coordination geometry. In the crystal, weak C\u2014H\u22efN and C\u2014H\u22efS hydrogen bonds link the complex mol\u00adecules into a two-dimensional network parallel to (010).The dinuclear title complex, [Ag For rel al. 2000; Lobana al. 2008; Isab et al. 2010.2(C6H7N2S)2(C18H15P)2] = 0.032wR(F2) = 0.088S = 1.045568 reflections266 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.54 e \u00c5\u22123\u0394\u03c1min = \u22120.29 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536812048210/bh2464Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "A weak intra\u00admolecular C\u2014H\u22efO hydrogen bond occurs between the DMBP and DMSO ligands. \u03c0\u2013\u03c0 stacking between pyridine rings [centroid\u2013centroid distances = 3.682\u2005(3) and 3.598\u2005(3)\u2005\u00c5] is observed in the crystal.In the title compound, [CdBr DOI: 10.1107/S1600536812028553/xu5571Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Pd atom is located on an inversion centre, and thus the asymmetric unit contains one half of the complex; the PdN2Cl2 unit is exactly planar. The dihedral angle between the PdN2Cl2 unit and quinoline ligand is 85.63\u2005(8)\u00b0. In the crystal, the complex mol\u00adecules are stacked into columns along the b axis. In the columns, several inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the six-membered rings are present, the shortest ring centroid\u2013centroid distance being 3.764\u2005(3)\u2005\u00c5 between pyridine rings.In the title complex, [PdCl II complex cis-[PtCl2(quinoline)2]\u00b70.25DMF, see: Davies et al. 2] = 0.040wR(F2) = 0.095S = 1.051577 reflections106 parametersH-atom parameters constrainedmax = 1.30 e \u00c5\u22123\u0394\u03c1min = \u22120.40 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 global. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal, there are aromatic \u03c0\u2013\u03c0 contacts between the pyridine rings [centroid\u2013centroid distances = 3.9577\u2005(13)\u2005\u00c5] and inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds.In the title compound, [Zn(NO DOI: 10.1107/S1600536811050227/bt5709Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The geometry around the RuII atom is pseudo-octa\u00adhedral, with two cis-binding PPh3 ligands and one chelating bidentate [Fc(O)PS2]2\u2212 ligand via two S atoms. The average Ru\u2014S and Ru\u2014P bond lengths are 2.434\u2005(1) and 2.398\u2005(1)\u2005\u00c5, and the Ru\u2014O and Ru\u2014C bond lengths are 2.157\u2005(3) and 1.826\u2005(4)\u2005\u00c5, respectively. In the crystal, pairs of O\u2014H\u22efO hydrogen bonds link adjacent mol\u00adecules into dimers.The structure of the title complex, [FeRu(C DOI: Click here for additional data file.10.1107/S1600536813014311/ds2231Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The whole repeating mol\u00adecular unit is generated by inversion symmetry. This leads to the formation of a looped-chain one-dimensional coordination polymer propagating along [110]. The dimethyl\u00adformamide (DMF) mol\u00adecules are linked to the chains via O\u2014H\u22efO hydrogen bonds. The chains are linked via N\u2014H\u22efO hydrogen bonds, forming two-dimensional networks parallel to (001). There are also a number of C\u2014H\u22efO inter\u00adactions present and a parallel slipped \u03c0\u2013\u03c0 inter\u00adaction. The latter involves inversion-related pyridine rings with a centroid\u2013centroid distance of 3.594\u2005(2)\u2005\u00c5 [normal distance = 3.3338\u2005(13) and slippage = 1.341\u2005\u00c5]. These inter\u00adactions lead to the formation of a three-dimensional structure.In the title coordination polymer, {[Cu(SO N,N\u2032-bis-(4-pyrid\u00adyl)isophthalamide ligand, see: Adarsh et al. (C18H14N4O2)(H2O)]\u00b7C3H7NO = 0.052wR(F2) = 0.124S = 1.075581 reflections339 parameters4 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.44 e \u00c5\u22123\u0394\u03c1min = \u22120.48 e \u00c5\u22123\u0394\u03c1CrystalClear used to solve structure: SHELXS97 I, global. DOI: Click here for additional data file.10.1107/S1600536813003413/su2555Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Two symmetry-related azide ligands bridge in \u03bc2-modes giving a Cu\u22efCu distance of 3.533\u2005(2)\u2005\u00c5. In the crystal, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds link the components into a three-dimensional network. In addition, there are weak inter\u00admolecular C\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions with centroid\u2013centroid distances ranging from 3.562\u2005(2) to 3.974\u2005(2)\u2005\u00c5. In the centrosymmetric dinuclear title complex, [Cu DOI: 10.1107/S1600536811045909/lh5362Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The SnIV atom of the one-dimensional coordination polymer is located on a mirror plane and is coordinated by two chelating oxalate ions with two rather different Sn\u2014O bond lengths of 2.150\u2005(1) and 2.425\u2005(1)\u2005\u00c5, and two t-butyl groups with Sn\u2014C bond lengths of 2.186\u2005(2) and 2.190\u2005(2)\u2005\u00c5. The coordination polyhedron around the SnIV atom is a distorted tetra\u00adgonal disphenoid. The centrosymmetric oxalate ion also has an asymmetric coordination geometry, as reflected by the two slightly different C\u2014O bond lengths of 1.242\u2005(2) and 1.269\u2005(2)\u2005\u00c5. The chains of the polymer propagate along the b-axis direction. Only van der Waals inter\u00adactions are observed between the chains.The title compound, [Sn(C For 2(C2O4)] = 0.015wR(F2) = 0.038S = 1.101548 reflections83 parametersH-atom parameters constrainedmax = 0.53 e \u00c5\u22123\u0394\u03c1min = \u22120.37 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I. DOI: 10.1107/S160053681400539X/cq2010Isup2.hklStructure factors: contains datablock(s) I. DOI: 990826CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "IV ion in the title complex, [PtBr4(C10H9N3)], is six-coordinated in a slightly distorted octa\u00adhedral environment by two pyridine N atoms from a chelating di-2-pyridyl\u00adamine (dpa) ligand and four Br\u2212 anions. The complex mol\u00adecule has mirror symmetry, with the PtIV atom, two Br atoms and the central N atom of the dpa ligand lying on the mirror plane. The dpa ligand is not planar, showing a dihedral angle of 34.7\u2005(2)\u00b0 between the pyridine rings. The complex mol\u00adecules are connected by inter\u00admolecular N\u2014H\u22efBr hydrogen bonds, forming chains along [001]. Inter\u00admolecular C\u2014H\u22efBr hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings [centroid\u2013centroid distance = 3.667\u2005(4)\u2005\u00c5] are also observed.The Pt DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The geometry around the MoVI atom is distorted octa\u00adhedral in each complex mol\u00adecule, supported by two oxide O atoms and the N2O2 donor atoms of the coordinating ligand. The dihedral angle between the benzene rings is 74.96\u2005(11)\u2005\u00c5 for mol\u00adecule A and 76.05\u2005(11)\u2005\u00c5 for mol\u00adecule B. In the crystal, the B mol\u00adecules are linked by pairs of C\u2014H\u22efCl hydrogen bonds, forming inversion dimers. The crystal structure is further stabilized by C\u2014H\u22ef\u03c0 inter\u00adactions. An inter\u00adesting feature of the crystal structure is a Cl\u22efCl contact [3.3748\u2005(18)\u2005\u00c5], which is shorter than the sum of the van der Waals radii of Cl atoms (3.50\u2005\u00c5).The asymmetric unit of the title compound, [Mo(C O2] = 0.027wR(F2) = 0.079S = 1.078626 reflections541 parametersH-atom parameters constrainedmax = 0.69 e \u00c5\u22123\u0394\u03c1min = \u22120.71 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SIR92 global, I. DOI: 10.1107/S160053681203807X/su2496Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The MnII atom is six-coordinated by 2\u2212 anions in a tridentate fashion and is at the centre of a distorted octa\u00adhedron formed by the MnO4N2 bonding set. In the crystal, various inter\u00admolecular inter\u00adactions between different moieties can be found, such as different kinds of hydrogen bonds, offset or slipped \u03c0\u2013\u03c0 [centroid\u2013centroid distances in the range 3.3704\u2005(12) to 3.8674\u2005(13)\u00c5] and C=O\u22ef\u03c0 [3.563\u2005\u00c5] inter\u00adactions, which lead to the formation of a three-dimensional supra\u00admolecular network. The asymmetric unit of the title compound, (C DOI: 10.1107/S1600536811036981/om2447Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "N,N\u2032-(o-phenyl\u00adene)bis\u00ad(pyridine-2-carboxamidato)]manganese(II)} ethanol hepta\u00adsolvate 3.5-hydrate, [Mn(C18H12N4O2)(C7H10N2)(NO)]8\u00b77C2H5OH\u00b73.5H2O, or 8[Mn(bpb)(DMAP)(NO)]\u00b77EtOH\u00b73.5H2O, is an unusual example of a structure with Z\u2032 = 8. The tetra\u00addentate bpb ligand, together with the nitrosyl and dimethyl\u00adamino\u00adpyridine ligands, gives rise to a distorted octa\u00adhedral coordination environment for the Mn(II) ion. The average Mn\u2014N(N=O) bond length is 1.631\u2005(13)\u2005\u00c5. The eight mol\u00adecules in the asymmetric unit differ mainly in the rotation of the DMAP pyridine plane with respect to a reference plane of the Mn and three N atoms, one of which is the N atom of the NO group. The dihedral angles between the normals to these planes range from a minimum of 28.0\u2005(2)\u00b0 to a maximum of 64.2\u2005(2)\u00b0. There are also some differences in O\u2014H\u22efO hydrogen bonding inter\u00adactions. For example, of the sixteen C=O acceptors, there are seven different inter\u00adactions with EtOH donors and two inter\u00adactions with H2O donors. The crystal studied was found to be a two-component twin, with a 179.9\u00b0 rotation about the real axis . Due to the presence of a superlattice and, consequently, the large number of weak reflections, the refinement utilized rigid solvate groups and isotropic displacement parameters for all except the Mn atoms. H atoms were not located for hydrate molecules.The structure of the title compound octa\u00adkis\u00ad{[4-(dimethyl\u00adamino)\u00adpyridine](nitros\u00adyl)[ DOI: 10.1107/S1600536811038669/wm2529Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The previous structure report [Spofford & Amma and 0.001\u2005\u00c5 (new)]. The asymmetric unit contains two formula units, with both CuI atoms coordinated by one terminal S atom and two bridging S atoms of thio\u00adurea ligands. This connectivity leads to polymeric [100] chains in the crystal. If very long contacts to nearby chloride ions [2.8687\u2005(9) and 3.1394\u2005(12)\u2005\u00c5] are considered to be bonding, then very distorted CuS3Cl tetra\u00adhedral coordination polyhedra arise. The crystal structure is consolidated by weak intra- and inter-chain N\u2014H\u22efS and N\u2014H\u22efCl hydrogen bonds.The structure of the polymeric title compound, [CuCl(CHAmma 1970. Acta Cr For the structure of a related thio\u00adurea salt, see: Zouihri 2012. For the4N2S)2] = 0.043wR(F2) = 0.081S = 1.154089 reflections245 parameters16 restraintsAll H-atom parameters refinedmax = 0.58 e \u00c5\u22123\u0394\u03c1min = \u22120.85 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536812004448/hb6598Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "It is built up by association of (2-(diphenyl\u00adphosphino)ferrocen\u00adyl)meth\u00adyl)dimethyl\u00adammonium cations and diphenyl\u00adphosphino dithio\u00adate anions. N\u2014H\u22efS, C\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions link the anions and cations. Each anion\u2013cation pair is linked two by two through C\u2014H\u22ef\u03c0 inter\u00adactions, forming pseudo dimers.2-(Diphenyl\u00adthio\u00adphosphino)dimethyl\u00adamino\u00admethyl\u00adferrocene is a key inter\u00admediate in the synthesis of various ferrocenyl ligands. During one such synthesis, the title compound, [Fe(C DOI: 10.1107/S1600536812009129/hp2031Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "VI atom in the title compound, [Mo(C16H14N2O4)O2(C2H6OS)], is distorted octa\u00adhedral. The phenolate O, imino N, oxide O from the enolized carbonyl group and one of the terminal O atoms form the equatorial plane; the axial positions are occupied by the other terminal O atom of the dioxidomolybdenum group and the donor O atom of DMSO. The O=Mo=O angle is 105.31\u2005(6)\u00b0. An intra\u00admolecular O\u2014H\u22efN hydrogen bond and weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds are present in the structure.The coordination geometry at the Mo O2(C2H6OS)] = 0.024 wR(F 2) = 0.120 S = 1.15 4556 reflections266 parametersH-atom parameters constrainedmax = 0.47 e \u00c5\u22123 \u0394\u03c1min = \u22121.68 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S1600536811020290/xu5215Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecules form an inversion dimer via a pair of weak C\u2014H\u22efO hydrogen bonds and a \u03c0\u2013\u03c0 stacking inter\u00adaction with a centroid\u2013centroid distance of 3.7401\u2005(12)\u2005\u00c5. Weak intra\u00admolecular C\u2014H\u22efBr inter\u00adactions and an intra\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adaction with a centroid\u2013centroid distance of 3.8118\u2005(15)\u2005\u00c5 are also observed.In the title compound, [MoBr DOI: 10.1107/S1600536811015881/is2705Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The benzimidazole ring system forms dihedral angles of 26.68\u2005(9) and 56.16\u2005(9)\u00b0 with the adjacent benzene ring and the methyl\u00adphenolate group benzene ring, respectively. In the crystal, mol\u00adecules are linked by N\u2014H\u22efCl hydrogen bonds into chains along [100]. Furthermore, weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions, in addition to \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances in the range 3.5826\u2005(13)\u20133.9681\u2005(13)\u2005\u00c5, are also observed.In the title mononuclear complex, [Zn(C Cl] = 0.042 wR(F 2) = 0.088 S = 1.03 5678 reflections249 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.64 e \u00c5\u22123 \u0394\u03c1min = \u22120.39 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXTL global, I. DOI: 10.1107/S1600536811030170/lh6598Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The SnIV ion is five-coordinated in a distorted trigonal\u2013bipyramidal geometry by two monodentate carboxyl\u00adate groups and three phenyl rings. The axial sites are occupied by the O atoms of two symmetry-related carboxyl\u00adate groups [O\u2014Sn\u2014O = 170.88\u2005(3)\u00b0]. The benzene ring of the 5-amino-2-nitro\u00adbenzoate ligand forms dihedral angles of 82.92\u2005(6), 81.10\u2005(6) and 83.54\u2005(6)\u00b0 with respect to the three phenyl rings. In the crystal, the chains are linked by inter\u00admolecular N\u2014H\u22efO and weak C\u2014H\u22efO inter\u00adactions into a three-dimensional network. The crystal structure is further stabilized by weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions.The title compound, [Sn(C For the al. 1987. 6H5)3(C7H5N2O4)] = 0.018 wR(F 2) = 0.046 S = 1.07 7981 reflections297 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.51 e \u00c5\u22123 \u0394\u03c1min = \u22120.54 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXTL global, I. DOI: 10.1107/S1600536811033332/lh5316Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the [FeBr4]\u2212 anion, the FeIII atom (3 symmetry) is four-coordinated in a distorted tetra\u00adhedral geometry. In the crystal, inter\u00admolecular C\u2014H\u22efBr hydrogen bonds and Br\u22ef\u03c0 inter\u00adactions [Br\u22efcentroid distances = 3.562\u2005(3) and 3.765\u2005(2)\u2005\u00c5] link the cations and anions, stabilizing the structure.In the [Fe DOI: 10.1107/S1600536811004181/hy2404Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, C\u2014H\u22efO and \u03c0\u2013\u03c0 inter\u00adactions connect the mol\u00adecules, forming double layers that stack along the 30H28N2O6, the pyran ring adopts a slightly distorted half-chair conformation and the pyrone ring adopts an envelope conformation, with the C atom bearing the carboxyl\u00adate group as the flap. The pyrazole ring [maximum deviation = 0.002\u2005(2)\u2005\u00c5] forms a dihedral angle of 13.2\u2005(1)\u00b0 with the attached benzene ring. The near-planar atoms of the pyran ring and the pyrazole ring are close to coplanar, the dihedral angles between their mean planes being 6.4\u2005(1)\u00b0. The dihedral angle between the pyrone ring and the benzene ring of the chromene unit is 10.7\u2005(1)\u00b0. The mol\u00adecular conformation is stabilized by an intra\u00admolecular C\u2014H\u22efO hydrogen bond, which generates an S(6) ring motif. In the crystal, C\u2014H\u22efO inter\u00adactions generate supra\u00admolecular chains propagating in [100] and these are connected into double layers that stack along the c-axis direction by weak \u03c0\u2013\u03c0 inter\u00adactions between pyrazole rings [centroid\u2013centroid distance = 3.801\u2005(1)\u2005\u00c5].In the title compound, C The pyrazole ring is approximately planar, with a maximum deviation of 0.002\u2005(2)\u2005\u00c5 for atom C2, and forms a dihedral angle of 13.2\u2005(1)\u00b0 with the attached benzene ring. The planar atoms of the pyran ring and the pyrazole ring are close to coplanar, the dihedral angles between their mean planes being 6.4\u2005(1)\u00b0. Moreover, the planar atoms of the pyrone ring and the benzene ring of the chromene unit are also almost coplanar, the dihedral angle between their mean planes being 10.7\u2005(1)\u00b0. The geometric parameters of the title mol\u00adecule agree well with those reported for similar structures ring motif. The crystal packing features C17\u2014H17\u22efO3 hydrogen bonds, which form a supra\u00admolecular chain along the a axis. This chain is connected into double layer that stacks along the c axis (Table\u00a01Cg is the centroid of the pyrazole N1/N2/C3/C1/C2 ring) by \u03c0\u2013\u03c0 inter\u00adactions, with Cg\u22efCgii = 3.801\u2005(1)\u2005\u00c5 .The mol\u00adecular conformation is stabilized by an intra\u00admolecular C19\u2014H19\u22efO1 hydrogen bond, which generates an et al., 2013a2,7.013,17]hepta\u00addeca-2(7),3,5,13\u2005(17),15-penta\u00adene-10-carboxyl\u00adate, (III) E)-methyl 2-[(2-formyl-6-meth\u00adoxy\u00adphen\u00adoxy)meth\u00adyl]-3-(4-meth\u00adoxy\u00adphen\u00adyl)acrylate and 3-methyl-1-phenyl-1H-pyrazol-5-one was placed in a round-bottomed flask and melted at 453\u2005K for 1\u2005h. After completion of the reaction as indicated by thin-layer chromatography, the crude product was washed with 5\u2005ml of an ethyl acetate and hexane mixture (1:49 ratio), which successfully provided the title compound as a colourless solid in 93% yield. Colourless blocks were obtained by slow evaporation of an ethyl acetate solution at room temperature.A mixture of (Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms. Owing to poor agreement, the reflections 100, 011 and 100 were omitted from the final cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814017929/hb7257sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S1600536814017929/hb7257Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814017929/hb7257Isup3.cmlSupporting information file. DOI: 962784CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Figure legend 1 is incorrect. The correct figure legend is:Figure 1. Effect of maggot secretions on the tPA-induced lysis of plasma clots (representative example). No secretions \u2014; 12.5 \u00b5g of secretions/mL \u2014 \u2014 \u2014; 25 \u00b5g of secretions/mL \u2013 \u2014 \u2013 \u2014; 50 \u00b5g of secretions/mL \u2013 \u2013 \u2013."} +{"text": "The title compound crystallized with two independent mol\u00adecules in the asymmetric unit. In the crystal, they are linked to one another, forming chains enclosing 22H17NO4, crystallizes with two independent mol\u00adecules (A and B) in the asymmetric unit. Each mol\u00adecule exists as an E isomer with C\u2014C=C\u2014C torsion angles of \u2212175.69\u2005(17) and \u2212178.41\u2005(17)\u00b0 in A and B, respectively. In mol\u00adecule A, the planes of the terminal benzene rings are twisted by an angle of 26.67\u2005(10)\u00b0, while the biphenyl unit is non-planar, the dihedral angle between the rings being 30.81\u2005(10)\u00b0. The dihedral angle between the nitro\u00adphenyl ring and the inner phenyl ring is 6.50\u2005(9)\u00b0. The corresponding values in mol\u00adecule B are 60.61\u2005(9), 31.07\u2005(8) and 31.05\u2005(9)\u00b0. In the crystal, mol\u00adecules are arranged in a head-to-head manner, with the 3-nitro\u00adphenyl groups nearly parallel to one another. The A and B mol\u00adecules are linked to one another via C\u2014H\u22efO hydrogen bonds, forming chains lying parallel to (-320) and enclosing R22(10) and R22(12) ring motifs. The meth\u00adoxy group in both mol\u00adecules is positionally disordered with a refined occupancy ratio of 0.979\u2005(4):0.021\u2005(4) for mol\u00adecule A and 0.55\u2005(4):0.45\u2005(4) for mol\u00adecule B.The title compound, C Each mol\u00adecule exists as an E isomer with the C17\u2014C16\u2014C15\u2014C14 and C39\u2014C38\u2014C37\u2014C36 torsion angles being \u2212175.69\u2005(17) and \u2212178.41\u2005(17)\u00b0, respectively. In mol\u00adecule A, the terminal benzene rings (C2\u2013C7) and (C17\u2013C22) are twisted by an angle of 26.67\u2005(10)\u00b0, while the biphenyl rings (C2\u2013C7 and C8\u2013C13) are non-planar, the dihedral angle being 30.81\u2005(10)\u00b0. The dihedral angle between rings (C8\u2013C13) and (C17\u2013C22) is 6.50\u2005(9)\u00b0. The corresponding dihedral angles in mol\u00adecule B are (C24\u2013C29 and C39\u2013C44) 60.61\u2005(9), (C30\u2014C35 and C24\u2013C29) 31.07\u2005(8) and (C30\u2013C35 and C39\u2013C44) 31.05\u2005(9)\u00b0.The title compound, Fig.\u00a01A and B lie head-to head almost parallel to one another. They are linked via C\u2014H\u22efO hydrogen bonds, forming chains lying parallel to -3-(biphenyl-4-yl)-1-prop-2-en-1-one -1--3-(2-methyl\u00adphen\u00adyl)prop-2-en-1-one -3-(3-nitro\u00adphen\u00adyl)prop-2-en-1-one and 6.50\u2005(9)\u00b0, respectively.A search of the Cambridge Structural Database and 3-nitro benzaldehyde in ethanol (25\u2005ml) in the presence of NaOH (10\u2005ml 30%) was heated in a water bath for 30\u2005min. and then allowed to cool. The solid that separated was filtered and recrystallized from ethanol. The yellow-coloured crystals of the title compound used for the X-ray diffraction study were grown by slow evaporation from acetone .Uiso(H) = 1.5Ueq(C) for methyl H atoms and = 1.2Ueq(C) for other H atoms. The refined occupancy ratios for the disordered meth\u00adoxy groups are 0.979\u2005(4):0.021\u2005(4) for atoms O1A/O1B and C1A/C1B in mol\u00adecule A and 0.55\u2005(4):0.45\u2005(4) for atoms O5A/O5B and C23A/C23B in mol\u00adecule B.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989014025110/su5021sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989014025110/su5021Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989014025110/su5021Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989014025110/su5021Isup4.cmlSupporting information file. DOI: 1034620CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The amide group is twisted by 31.30\u2005(16)\u00b0 with respect to the pyridine ring. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO, O\u2014H\u22efS and N\u2014H\u22efS hydrogen bonds into a three-dimensional supra\u00admolecular network. Weak \u03c0\u2013\u03c0 stacking is observed between parallel pyridine rings of adjacent mol\u00adecules, the centroid\u2013centroid distance being 3.8270\u2005(19)\u2005\u00c5.In the title compound, [Co(NCS) DOI: 10.1107/S1600536814011453/xu5790Isup2.hklStructure factors: contains datablock(s) I. DOI: 850080CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the structure of the naphtho\u00adquinone derivative 2-hy\u00addroxy-3-(2-methyl\u00adprop-1-en-1-yl)naphthalene-1,4-dione, the mol\u00adecules form a centrosymmetric cyclic dimer through inter\u00admolecular O\u2014H\u22efO hydrogen bonds which, together with inter\u00admolecular C\u2014H\u22efO hydrogen bonds and weak \u03c0\u2013\u03c0 ring inter\u00adactions, give rise to an overall two-dimensional structure. 14H12O3, the substituent side chain, in which the H atoms of both methyl groups are disordered over six equivalent sites, lies outside of the plane of the naphthalene\u00addione ring. The ring-to-chain C\u2014C\u2014C\u2014C torsion angles are 50.7\u2005(3), \u2212176.6\u2005(2) and 4.9\u2005(4)\u00b0. An intra\u00admolecular meth\u00adyl\u2013hy\u00addroxy C\u2014H\u22efO hydrogen bond is present. In the crystal, mol\u00adecules are primarily connected by inter\u00admolecular O\u2014H\u22efO hydrogen bonds, forming a centrosymmetric cyclic dimer motif [graph set R22(10)]. Also present is a weak inter\u00admolecular C\u2014H\u22efO hydrogen bond linking the dimers and a weak \u03c0\u2013\u03c0 ring inter\u00adaction [ring centroid separation = 3.7862\u2005(13)\u2005\u00c5], giving layers parallel to (10-3).In the structure of the title compound, C Also present in the structure is a weak inter\u00admolecular C7\u2014H\u22efO2ii hydrogen bond [3.339\u2005(3)\u2005\u00c5], linking the dimers and a weak \u03c0\u2013\u03c0 ring inter\u00adaction between the benzene and quinone ring moieties of the parent ring system , giving layers parallel to = 1.2Ueq(C). Rotational disorder was identified in the hydrogen atoms of the methyl carbon atoms C12 and C22 and these were included in the refinement over six equivalent 60\u00b0 sites with 50% occupation, with C\u2014H = 0.96\u2005\u00c5 and Uiso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015024755/zs2357sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015024755/zs2357Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015024755/zs2357Isup3.cmlSupporting information file. DOI: 1444109CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The ligand selection of bidentate \u03b2\u2010diketonates is shown to be key to isolating a cyclic trimer. Additional reaction of \u03b2\u2010diketonates with diethyl zinc were spectroscopically characterized as compounds of the type [{Zn(Et)(\u03b2\u2010diketonate)}n] (\u03b2\u2010diketonate=OC(Me)CHC(Me)O, 2, OC(OtBu)CHC(Me)O, 3). Further studies have shown that selective oxidation of these species produces cubanes of the general formula [{Zn(OC(R)CHC(Me)O)2Zn(Et)OEt}2] , allowing a high oxygen content whilst remaining structurally suitable for use as precursors. The successful deposition of thin films of zinc oxide through aerosol\u2010assisted chemical vapor deposition (AACVD), using a novel precursor, is described and fully characterized.A conventional solution\u2010based route to a cyclic trimeric organozinc compound [{Zn(Et)( Studies involving zinc alkoxides were first reported by Frankland in 18492Zn compounds through the contributions of Lewi\u0144ski et\u2005al. and a handful of other groups.azol)]2[EtZn(azol)]2, bridged by peroxide groups. Interestingly, this work was the first to \u2018suggest the presence of a trimer as the predominate species in solution\u2019 of the precursor aggregate, namely, [EtZn(azol)]n.The widely accepted belief that the oxidation of zinc alkyls is so fast, selectivity is not possible, was first dispelled by the structural characterization of a novel zinc alkylperoxide.It is known that the structure of organozinc compounds can be enormously varied, including multinuclear rings3 \u20102\u2010(dimethylaminomethyl)\u2010iso\u2010propoxide), whilst not crystallographically characterized, has a cyclic, not a roof\u2010like, trimeric structure (thf)}2] under mild conditions to give [{tBuZnOtBu}3]. This cluster was reported to have two distinct types of organozinc centers: two four\u2010coordinate zinc centers linked by two \u03bc3\u2010 and one \u03bc2\u2010oxygen atoms, and one three\u2010coordinate zinc center binding to two \u03bc2\u2010oxygen atoms. It was found to be a metastable product, as solution\u2010based attempts to form the compound resulted solely in the tetramer. Later in 2012, it was shown that by using a more rigid quinoline ligand, thus moving from a ZnCO3 coordination environment to ZnCNO2, a trimer could be isolated and was structurally characterized as [tBuZn(q)]3 (q=8\u2010hydroxyquinoline).In 2010, the first crystallographically characterized organozinc \u2018dimeric aggregate\u2019 with a Zn3 , in which the use of bidentate \u03b2\u2010diketonate ligands facilitated crystallization. Similar reactions of diethyl zinc with \u03b2\u2010diketonates yielded compounds of the type [{Zn(Et)(\u03b2\u2010diketonate)}n], which were spectroscopically characterized. In addition, selective reaction of these compounds with oxygen yields distorted cubanes of the general formula [{Zn(\u03b2\u2010diketonate)2Zn(Et)OEt}2]. These compounds exhibit a similar central motif to that reported previously.Herein, we report the first instance of a conventional solution\u2010based synthetic route to a structurally characterized cyclic trimeric organozinc compound with the coordination environment ZnCO3 Figure\u2005\u2009a, [{Zn(\u03b2\u2010diketonate [{Zn(Et)(OC(OMe)CHC(Me)O)}3]\u2005(1) was isolated and structurally characterized; furthermore, compounds of stoichiometry [{Zn(Et)(OC(R)CHC(Me)O)}n], where R=Me\u2005(2) and OtBu\u2005(3) (Scheme\u20051\u20133 were isolated from the equimolar reaction of Et2Zn with the respective carbonyl in high yield (>90\u2009%) and characterized by using spectroscopic techniques. Although 2 and 3 formed crystalline solids, none were of suitable quality for crystallographic analysis.The trimeric Me)O)}3]\u2005 was isol1 crystallizes in the monoclinic space group P21/n with three zinc centers, all exhibiting a distorted tetrahedral coordination \u2013O(O(1):2.0489(9)\u2005\u00c5, Zn(2)\u2013O(4):2.0247(9)\u2005\u00c5, Zn(3)\u2013O(7):2.0367(9)\u2005\u00c5] and the longer bridging Zn\u2212O lengths , which is comparable to similar structures found in the literature.1 results largely from the constraints of the central Zn3O3 ring and the three outer ZnO2C3 rings formed from coordination to the bidentate ligands.Compound\u2005tBuZnOtBu}3] \u2013O(7), 3.042\u2005\u00c5, which is considerably larger than the comparable distance in [{tBuZnOtBu}3] (2.164\u2005\u00c5),1.For comparison with the previously reported \u2018dimeric aggregate\u2019 [{] Figure\u2005\u2009c, the ctBu}3] 2.4\u2005\u00c5,16 pr\u03b2\u2010diketonate ligand facilitates the isolation of a novel type of cyclic trimeric organozinc complex\u2005(1) with a coordination environment of ZnCO3, as confirmed by crystallographic characterization. The monodentate tBu groups used by Lewi\u0144ski resulted in a metastable roof\u2010like trimer (or aggregated dimer), whereas the flexible amino alcoholates utilized by Molloy and co\u2010workers yielded oils, not solids, although NMR studies did suggest a cyclic trimeric center.The above data indicate that the use of a rigid bidentate 1\u20133 were explored.The rationale behind this ligand choice firstly lies in the successful isolation of a novel structural type, but also in increasing the oxygen loading of the compounds (as zinc precursors can lead to oxygen\u2010deficient thin films).2Zn(Et)OEt}2] (R=OMe\u2005(4), Me\u2005(5), OtBu\u2005(6)) were synthesized through the controlled addition of O2 to solutions of 1\u20133 (Scheme\u2005\u03bc2\u2010OEt group in accordance with previous reports.4\u20136 were isolated in high yields (>80\u2009%) and characterized by using spectroscopic techniques. Compounds\u20055 and 6 both crystallized out of concentrated solutions held at \u221218\u2009\u00b0C as centrosymmetric clusters in the triclinic P\u03041 space group with four zinc centers; two of which are unique and two are symmetrically generated about an inversion center .\u2005(2 and 3). Through selective oxidation, we have subsequently shown the synthesis of [{Zn(OC(R)CHC(Me)O)2Zn(Et)OEt}2]\u2005(4\u20136). X\u2010ray crystallography of 5 and 6 revealed these complexes to have a face\u2010shared, corner\u2010removed, inversion\u2010related, bis\u2010heterocubane central motif. Of additional importance is that these materials can be used as precursors to thin films of zinc oxide; as exemplified in the successful deposition of hexagonal wurtzite zinc oxide thin films. We believe these results provide a new route, through careful ligand selection, to cyclic organozinc trimers, whose rarity was commented on by Power and co\u2010workers back in 1991.In summary, we have demonstrated the solution\u2010based synthesis of the first structurally characterized cyclic trimer with a Zn1, 5, and 6 can be found in the Supporting Information.Crystallographic/refinement data for compounds\u2005m in hexanes) was obtained from Sigma Aldrich and used as supplied. Acetylacetone, methyl acetoacetate, and tert\u2010butyl acetoacetate were obtained from Sigma Aldrich, degassed, and stored over activated molecular sieves.All manipulations were performed under a dry dinitrogen atmosphere by using standard Schlenk techniques. Hexane was stored in an alumina column and dried with anhydrous engineering equipment. Diethylzinc solution . Mass spectroscopy was performed on a Thermo Finnigan MAT900 XP operating in electron impact and chemical ionization modes. Single\u2010crystal X\u2010ray diffraction datasets were collected on a SuperNova Atlas diffractometer by using either monochromated Cu K\u03b1 radiation (\u03bb=1.54184\u2005\u00c5) or monochromated Mo K\u03b1 (\u03bb=0.71073\u2005\u00c5).m solution in hexanes, 9.76\u2005mmol) was added to dry hexane (5\u2005mL) at \u221278\u2009\u00b0C. Dry methyl acetoacetate was added dropwise to the solution and stirred at RT for 48\u2005h. Hexane was fully removed in\u2005vacuo, resulting in an off\u2010white solid. Off\u2010white crystals grew from a concentrated solution held at \u221218\u2009\u00b0C. Yield: 1.86\u2005g, 91\u2009%; 1H\u2005NMR (600\u2005MHz) \u03b4 (C6D6): 0.55 , 1.47 , 1.90 , 3.27 , and 4.95\u2005ppm ; 13C{1H} NMR (600\u2005MHz) \u03b4 (C6D6): 1.0 (CH2CH3), 12.5 (CH2CH3), 27.5 (CCH3), 51.1 (OCH3), 89.3 (CCHC), 174.0 (COCH3), and 184.4\u2005ppm (CCH3); anal. calcd. for C21H36O9Zn3: C 40.12, H 5.77; found: C 40.21, H 5.89.Diethylzinc was added to dry hexane (5\u2005mL) at \u221278\u2009\u00b0C. Dry acetylacetone was added dropwise to the solution and stirred at RT for 48\u2005h. Hexane was fully removed in\u2005vacuo, resulting in an off\u2010white solid. Yield: 1.71\u2005g, 90\u2009%; 1H\u2005NMR (600\u2005MHz) \u03b4 (C6D6): 0.57 , 1.48 , 1.79 , and 5.03\u2005ppm ; 13C{1H} NMR (600\u2005MHz) \u03b4 (C6D6): 1.0 (CH2CH3), 12.4 (CH2CH3), 28.3 (CCH3), 102.0 (CCHC), and 193.1\u2005ppm (CCH3); anal. calcd. for C21H36O6Zn3: C 43.44, H 6.25; found: C 43.09, H 6.77.Diethylzinc was added to dry hexane (5\u2005mL) at \u221278\u2009\u00b0C. Dry tert\u2010butyl acetoacetate was added dropwise to the solution and stirred at RT for 48\u2005h. Hexane was fully removed in\u2005vacuo, resulting in an off\u2010white solid. Yield: 2.25\u2005g, 91\u2009%; 1H\u2005NMR (600\u2005MHz) \u03b4 (C6D6): 0.37 , 1.31 3), 1.33 , 1.94 , and 4.90\u2005ppm ; 13C{1H} NMR \u03b4 (600\u2005MHz) (C6D6): 3.4 (CH2CH3), 11.7 (CH2CH3), 27.6 (CCH3), 28.4 (C(CH3)3), 80.9 C(CH3)3, 91.0 (CCHC), 173.8 (COC(CH3)3), and 183.2\u2005ppm (CCH3); anal. calcd. for C30H54O9Zn3: C 47.73, H 7.21; found: C 46.93; H 6.97.Diethylzinc was added to a solution of 1 and stirred for 15\u2005min at \u221278\u2009\u00b0C. The flask was purged with N2 and stirred at room temperature for 24\u2005h. Off\u2010white crystals formed from a concentrated solution held at \u221218\u2009\u00b0C. Yield: 1.85\u2005g, 87\u2009%; 1H\u2005NMR (600\u2005MHz) \u03b4 (C6D6): 0.62 , 1.33 , 1.60 , 1.90 , 3.33 , 3.87 , and 4.99\u2005ppm ; 13C{1H} NMR (600\u2005MHz) \u03b4 (C6D6): 2.2 (ZnCH2CH3), 12.6 (ZnCH2CH3), 19.7 (OCH2CH3), 27.3 (CCH3), 50.6 (OCH3), 61.3 (OCH2CH3), 87.0 (CCHC), 173.6 (COCH3), and 183.3\u2005ppm (CCH3); anal. calcd. for C28H48O14Zn4: C 38.64, H 5.56; found: C 38.45, H 5.64. MS: m/z [M\u2212Zn2O5C13H27]+\u22c5: 474.87; [M\u2212Zn3O8C18H34]+\u22c5: 294.97.O2 (5\u2005mL) was added to a solution of 2 and stirred for 15\u2005min at \u221278\u2009\u00b0C. The flask was purged with N2 and stirred at room temperature for 24\u2005h. Off\u2010white crystals formed from a concentrated solution held at \u221218\u2009\u00b0C. Yield: 1.66\u2005g, 84\u2009%; 1H\u2005NMR (600\u2005MHz) \u03b4 (C6D6): 0.69 , 1.42 , 1.61 , 1.79 , 3.80 , and 5.06\u2005ppm ; 13C{1H} NMR (600\u2005MHz) \u03b4 (C6D6): 2.7 (ZnCH2CH3), 12.9 (ZnCH2CH3), 20.1 (OCH2CH3), 28.1 (CCH3), 61.1 (OCH2CH3), 100.6 (CCHC), and 193.1\u2005ppm (CCH3); anal. calcd. for C28H48O10Zn4: C 41.71, H 6.00; found: C 42.06, H 5.92. MS: m/z [M+C3H5]+\u22c5: 847.04; [M\u2212C8H20]+\u22c5: 691.11; [M\u2212Zn2O4C13H27]+\u22c5: 426.91; [M\u2212Zn3O6C18H34]+\u22c5: 263.00.O2 (5\u2005mL) was added to a solution of 3 and stirred for 15\u2005min at \u221278\u2009\u00b0C. The flask was purged with N2 and stirred at room temperature for 24\u2005h. Off\u2010white crystals formed from a concentrated solution held at \u221218\u2009\u00b0C. Yield: 2.09\u2005g, 83\u2009%; 1H\u2005NMR (600\u2005MHz) \u03b4 (C6D6): 0.58 (q (br), 4\u2009H, ZnCH2CH3), 1.44 3), 1.51 (t (br), 6\u2009H, OCH2CH3), 1.66 , 1.92 , 4.02 , and 4.96\u2005ppm ; 13C{1H} NMR (600\u2005MHz) \u03b4 (C6D6): 1.8 (ZnCH2CH3), 13.1 (ZnCH2CH3), 19.8 (OCH2CH3), 27.4 (CCH3), 28.7 (C(CH3)3), 61.4 (OCH2CH3), 79.7 (C(CH3)3), 88.7 (CCHC), 173.4 (COC(CH3)3), and 182.1\u2005ppm (CCH3); anal. calcd. for C40H72O14Zn4: C 46.26, H 6.99; found: C 46.43, H 7.27. MS: m/z [M\u2212Zn2O5C16H33]+\u22c5: 601.05; [M\u2212Zn3O8C24H46]+\u22c5: 379.04.O2. [(Zn(OC(Me)CHC(Me)O)2Zn(Et)O(Et))2]\u2005(5) was dissolved in dry toluene (30\u2005mL) under N2 and stirred for 10\u2005min. Thin films were deposited by using optimal conditions of a N2 flow rate of 1.2\u2005L\u2009min\u22121, a substrate temperature of 450\u2009\u00b0C, and annealing in air for 5\u2005h at 600\u2009\u00b0C. XRD was performed using a Bruker D8 Discover X\u2010ray diffractometer by using monochromatic Cu K\u03b11 and Cu K\u03b12 radiation of wavelengths 1.54056 and 1.54439\u2005\u00c5, respectively, emitted in an intensity ratio of 2:1 with a voltage of 40\u2005kV and a current of 40\u2005mA. SEM was performed by using a Philips XL30 FEG operating in plan and cross\u2010section mode with an electron beam accelerating energy of 30\u2005kV. XPS surface and depth profiling was performed by using a Thermo Scientific K\u2010Alpha XPS system with monochromatic Al K\u03b1 radiation at 1486.6\u2005eV as the X\u2010ray source. Etching was achieved by using an Ar\u2010ion etch beam at 1\u2005KeV with a current of 1.51\u2005\u03bcA. CasaXPS software was used to analyze the data with binding energies referenced to an adventitious C\u20091s peak at 284.8\u2005eV. UV/Vis/NIR transmission spectra were recorded by using a PerkinElmer Lambda 950 spectrometer in the range of 300\u20131400\u2005nm with an air background.Films were deposited onto float\u2010glass substrates with a 25\u2005nm barrier layer of crystalline SiOAs a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "E,E)-2\u2032,4\u2032-di\u00adhydroxy\u00adaceto\u00adphenone azine mol\u00adecule is located on an inversion centre and linked with di\u00admethyl\u00adformamide solvent mol\u00adecules via O\u2014H\u22efO hydrogen bonds.The approximately planar -2\u2032,4\u2032-di\u00adhydroxy\u00adaceto\u00adphenone azine mol\u00adecule is centrosymmetric, the mid-point of the N\u2014N bond being located on an inversion centre. All the non-H atoms of the azine mol\u00adecule are approximately coplanar, the maximum deviation being 0.017\u2005(2)\u2005\u00c5. An intra\u00admolecular O\u2014H\u22efN hydrogen bond occurs between the azine N atom and the hy\u00addroxy group. In the crystal, azine and di\u00admethyl\u00adformamide solvent mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds.In the title compound {systematic name: 4,4\u2032-bis\u00ad}, C Fluorescence properties of hydrazones have been reported -2\u2032,4\u2032-di\u00adhydroxy\u00adaceto\u00adphenone azine di\u00admethyl\u00adformamide disolvate, (I)Although 2\u2032,4\u2032-di\u00adhydroxy\u00adaceto\u00adphenone azine has been pre\u00adpared and studied as a fluorescent probe, its structure has not been reported. As a part of our studies on synthesis and structural peculiarities of Schiff base ligands derived from 2\u2032,4\u2032-di\u00adhydroxy\u00adaceto\u00adphenone and hydrazine, we determined the structure of the title compound, -2\u2032,4\u2032-di\u00adhydroxy\u00adaceto\u00adphenone azine and one dimethylformamide (DMF) mol\u00adecule. The complete azine mol\u00adecule is centrosymmetric and exists in an E,E configuration with respect to the two C=N bonds. The N1\u2014C2 bond length of 1.301\u2005(3)\u2005\u00c5 shows double-bond character. The C\u2014O bond lengths [1.349\u2005(3) and 1.358\u2005(3)\u2005\u00c5] are comparable with similar bonds in related structures are essentially planar, with a mean deviation of 0.0024\u2005\u00c5. Each hy\u00addroxy group is nearly coplanar with its attached benzene ring; the r.m.s. deviation is 0.0045\u2005\u00c5 for the seven non-H atoms. Intra\u00admolecular O\u2014H\u22efN hydrogen bonds exist in the azine mol\u00adecule , inter\u00admolecular O\u2014H\u22efO hydrogen bonds exist between azine mol\u00adecules and DMF mol\u00adecules Table\u00a01.E,E)-2,2\u2032-di\u00adphenol -4,4\u2032-di\u00adchloro-2,2\u2032-diphenol -3,3\u2032-dieth\u00adoxy-2,2\u2032-diphenol -4,4\u2032-dimeth\u00adoxy-2,2\u2032-diphenol : \u03b4 13.59 , 10.14 , 7.58\u20137.61 , 6.37\u20136.41 , 6.30\u20136.31 , 3.34 .A mixture of 2\u2032,4\u2032-di\u00adhydroxy\u00adaceto\u00adphenone , hydrazine sulfate and tri\u00adethyl\u00adamine in ethanol (40\u2005ml) was heated under reflux for 24\u2005h. After cooling, the precipitate was filtrated and washed with water to afford a yellow solid. Crystals of the title compound suitable for X-ray diffraction were obtained by slow evaporation of a solution of the solid in DMF at room temperature for 5\u2005d . Uiso(H) = 1.2Ueq(C) for aromatic H atoms or 1.5Ueq for methyl and hy\u00addroxy groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016003686/xu5885sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016003686/xu5885Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016003686/xu5885Isup3.cmlSupporting information file. DOI: 1457201CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Pd\u2014N bond distances are 2.022\u2005(2) and 2.027\u2005(2)\u2005\u00c5, the Pd\u2014Cl bond distances are 2.2880\u2005(7) and 2.2833\u2005(7)\u2005\u00c5, and the ligand bite angle is 80.07\u2005(9)\u00b0. The dimethyl sulfoxide and water mol\u00adecules form linear chains along [100] by O\u2014H\u22efO and O\u2014H\u22efS hydrogen bonds, generating eight- and 12-membered rings. C\u2014H\u22efCl inter\u00adactions link the chains, forming a three-dimensional arrangement. In addition, the 4,4-di-tert-butyl-2,2\u2032-bi\u00adpyridine ligand exhibits \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.8741\u2005(15) and 3.8353\u2005(15)\u2005\u00c5]. The DMSO solvent is disordered and was refined with an occupancy ratio of 0.866\u2005(3):0.134\u2005(3).The title compound, [PdCl DOI: 10.1107/S1600536814009453/pj2010Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, forming chains propagating along the 23H20BrNO3S, the \u2013(CH2)4\u2013 atoms are positionally disordered [occupancy ratio = 0.753\u2005(6):0.247\u2005(6)]. The ring has a half-chair conformation for both the major and minor components. The dihedral angles between the mean plane of the thio\u00adphene ring and those of the benzene and phenyl rings are 35.2\u2005(4) and 57.7\u2005(3)\u00b0, respectively. The planes of the two aryl rings are twisted with respect to each other by 86.4\u2005(6)\u00b0. In the mol\u00adecule, there is an O\u2014H\u22efN hydrogen bond forming an S(6) ring motif. In the crystal, mol\u00adecules are linked via C\u2014H\u22efO hydrogen bonds, forming chains parallel to [100].In the cyclo\u00adhexene ring of the title compound, C Reviews on the synthesis and properties of these compounds have been reported (Sabnis A/C4\u2013C7/C7A) and (C3A/C44\u2013C47/C7A) adopt a half-chair conformation. The mean plane of the major component (C3A/C4\u2013C7/C7A) is slightly twisted from the mean plane of the thio\u00adphene ring (S1/C2/C3/C3A/C7A) by 5.18\u2005(14)\u00b0. The dihedral angles between the mean plane of the thio\u00adphene ring and the benzene (C21\u2013C26) and phenyl (C31\u2013C36) rings are 35.2\u2005(4) and 57.7\u2005(3)\u00b0, respectively. The two aryl rings are normal to each other, making a dihedral angle of 86.4\u2005(6)\u00b0. In the mol\u00adecule there is an O\u2014H\u22efN hydrogen bond forming an S(6) ring motif methanone derivatives gave eight hits, which include five structures closely related to the title compound. These include [2-[(2-hy\u00addroxy\u00adbenzyl\u00adidene)amino](phen\u00adyl)methanone (I)et al., 2014aet al., 2014bb]thio\u00adphen-3\u00adyl](phen\u00adyl)methanone (III) -4,5,6,7-tetra\u00adhydro\u00adbenzo[b]thio\u00adphen-3-yl](phen\u00ad\u00adyl)methanone (IV) thio\u00adphene-3-yl](phen\u00adyl)methanone (V) thiophen-3-yl)-phenyl\u00admethanone in 10\u2005ml of methanol an equimolar amount of 5-bromo-2-hy\u00addroxy-3-meth\u00adoxy\u00adbenzaldehyde was added with constant stirring. The mixture was refluxed for 6\u2005h. A yellowish brown precipitate was obtained. Completion of the reaction was confirmed by thin layer chromatography. The precipitate obtained was filtered and dried at room temperature overnight. The solid was then recrystallized using a 1:1 solution of aceto\u00adnitrile and di\u00adchloro\u00admethane, giving colourless block-like crystals.To a solution of = 1.5Ueq for the hydroxyl and methyl H atoms, and = 1.2Ueq(C) for other H atoms. A single weak outlier reflection I. DOI: 10.1107/S2056989015000195/su5055Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015000195/su5055Isup3.cmlSupporting information file. DOI: 1042320CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "These inter\u00adactions exhibit a parallel displaced \u03c0\u2013\u03c0 stacking mode. Additional weak C\u2014H\u22ef\u03c0-ring and C\u2014H\u22efN and C\u2014H\u22efF inter\u00adactions are found, leading to a three-dimensional architecture. The RuII atom is coordinated in a distorted octa\u00adhedral geometry. The counter-charge is provided by two hexa\u00adfluorido\u00adphosphate anions and the asymmetric unit is completed by three aceto\u00adnitrile solvent mol\u00adecules of crystallization. Four F atoms of one PF6\u2212 anion are disordered over three sets of sites with occupancies of 0.517\u2005(3):0.244\u2005(3):0.239\u2005(3). Two aceto\u00adnitrile solvent mol\u00adecules are highly disordered and their estimated scattering contribution was subtracted from the observed diffraction data using the SQUEEZE option in PLATON .In the title compound, Spek 2009. Acta Cr II complexes with tape and bpy-type ligands, see: Brietzke et al. 2(C16H8N4)](PF6)2\u00b73C2H3N = 0.047wR(F2) = 0.124S = 0.948918 reflections743 parameters363 restraintsH-atom parameters constrainedmax = 0.51 e \u00c5\u22123\u0394\u03c1min = \u22120.54 e \u00c5\u22123\u0394\u03c1X-AREA global, I. DOI: 10.1107/S1600536814011969/tk5315Isup2.hklStructure factors: contains datablock(s) I. DOI: 1004679CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked 25H25N3O, comprises a 2-amino\u00adpyridine ring fused with a cyclo\u00adheptane ring, which adopts a chair conformation. The central pyridine ring (r.m.s. deviation = 0.013\u2005\u00c5) carries three substituents, viz. a benzyl\u00adamino group, a meth\u00adoxy\u00adphenyl ring and a carbo\u00adnitrile group. The N atom of the carbo\u00adnitrile group is significantly displaced [by 0.2247\u2005(1)\u2005\u00c5] from the plane of the pyridine ring, probably due to steric crowding involving the adjacent substituents. The phenyl and benzene rings are inclined to one another by 58.91\u2005(7)\u00b0 and to the pyridine ring by 76.68\u2005(7) and 49.80\u2005(6)\u00b0, respectively. In the crystal, inversion dimers linked by pairs of N\u2014H\u22efNnitrile hydrogen bonds generate R22(14) loops. The dimers are linked by C\u2014H\u22ef\u03c0 and slipped parallel \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.6532\u2005(3)\u2005\u00c5] into a three-dimensional structure.The title compound, C The cyclo\u00adheptane ring adopts a chair conformation with puckering parameters Q2 = 0.4634\u2005(15)\u2005\u00c5, \u03d52 = 304.24\u2005(18)\u00b0 and Q3 = 0.6481\u2005(16)\u2005\u00c5 and \u03d53 = 284.37\u2005(12)\u00b0. The phenyl (C22\u2013C27) and benzene (C31\u2013C36) rings are inclined to one another by 58.91\u2005(7)\u00b0 and to the pyridine (N3/C2\u2013C6) ring by 76.68\u2005(7) and 49.80\u2005(6)\u00b0, respectively. The N atom of the carbo\u00adnitrile group, N1, is significantly displaced by 0.2247\u2005(1)\u2005\u00c5 from the plane of the pyridine ring, perhaps due to steric crowding. The shortening of the C\u2014N distance [C5\u2014N3 = 1.3390\u2005(14)\u2005\u00c5] and the opening of the N3\u2014C5\u2014C4 angle to 124.47\u2005(10)\u00b0 may be attributed to the size of the substituent at C1, and correlates well with the values observed in a similar structure \u2005\u00c5 is shorter than the average conjugated C\u2014N single bond, 1.370\u2005(1)\u2005\u00c5, found in the Cambridge Structural Database \u2005\u00c5 is significantly shorter than the O1\u2014C37 distance of 1.410\u2005(2)\u2005\u00c5. An enlarge\u00adment of bond angle [C33\u2014C34\u2014O1 = 124.34\u2005(13)\u00b0] on one side and a narrowing of bond angle [C35\u2014C34\u2014O1 = 116.29\u2005(12)\u00b0] on the other side of the benzene ring may be due to the steric repulsion between the aromatic rings and the methyl group, as found in a similar structure , normal distance = 3.5920\u2005(5), slippage = 0.667\u2005\u00c5; Cg1 is the centroid of the N3/C2\u2013C6 ring; symmetry code: (i) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01], as shown in Fig.\u00a02In the crystal, mol\u00adecules are linked r Table\u00a01. They arH-cyclohepta\u00ad[b]pyridine -4-(4-meth\u00adoxy\u00adphen\u00adyl)-6,7,8,9-tetra\u00adhydro-5p-TSA (1.0\u2005mmol) was added. The reaction mixture was heated under reflux for 2\u20133\u2005h. On completion of the reaction, checked by thin-layer chromatography (TLC), the mixture was poured into crushed ice and extracted with ethyl acetate. The excess solvent was removed under vacuum and the residue was subjected to column chromatography using petroleum ether/ethyl acetate mixture (97:3 v/v) as eluent to afford pure product. The product was recrystallized from ethyl acetate, affording colourless crystals of the title compound. .A mixture of cyclo\u00adhepta\u00adnone (1\u2005mmol), 4-meth\u00adoxy aldehyde (1\u2005mmol) and malono\u00adnitrile (1\u2005mmol) and benzyl\u00adamine (1mmol) was taken in ethanol (10\u2005ml) to which Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq for other H atoms. The DELU restraint was applied.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814024878/su5014sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S1600536814024878/su5014Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814024878/su5014Isup3.cmlSupporting information file. DOI: 1033842CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The naphthyl moieties are close to perpendicular [dihedral angle = 72.08\u2005(5)\u00b0]. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds, generating layers which surround channels occupied by disordered guest mol\u00adecules.In the solid-state, bis- 44H44N2O8, iminium\u00adyl]meth\u00adyl}-3,3\u2032-dimethyl-2,2\u2032-bi\u00adnaphthalene-7,7\u2032-diolate) has been obtained by the addition of p-anizidine to gossypol dissolved in di\u00adchloro\u00admethane. In the solid state, the title compound exists in the enamine or quinoid form. The two naphthyl moieties are inclined to one another by 72.08\u2005(5)\u00b0. The pendant phenyl rings are inclined at 22.26\u2005(14) and 23.86\u2005(13)\u00b0 to the corresponding naphthyl rings. In the crystal, mol\u00adecules are incorporated into layers through inversion-related pairs of O\u2014H\u22efO inter\u00adactions [graph sets R22(20) and R22(10)] and translation-related O\u2014H\u22efO inter\u00adactions [graph set C(15)]. The packing of these layers in the crystal structure gives rise to channels in the [011] direction, with hydro\u00adphobic inter\u00adactions occurring between adjacent layers. The channels are 5\u20137\u2005\u00c5 wide, and the void volume of each cell is 655\u2005\u00c53, corresponding to 26.6% of the cell volume. Disordered guest mol\u00adecules, probably solvent and water mol\u00adecules, occupy these voids of the crystal; their contribution to the scattering was removed with the SQUEEZE routine of PLATON .The title compound, CSpek 2015. Acta CrSpek 2009. Acta Cr However, bis-p-anizidinegossypol crystallizes in a triclinic and 1.476\u2005(4)\u2005\u00c5. In addition, the N1\u2014C22 [1.332\u2005(4)\u2005\u00c5] and N2\u2014C27 [1.319\u2005(4)\u2005\u00c5] bonds are shorter than N1\u2014C31 [1.433\u2005(4)\u2005\u00c5] and N2\u2014C38 [1.441\u2005(4)\u2005\u00c5], respectively. Contrarily, C7=O3 [1.289\u2005(3)\u2005\u00c5] and C17=O7 [1.275\u2005(3)\u2005\u00c5] bonds are longer than typical standard values.The mol\u00adecule consists of four ring systems, two of which are naphthalene ring systems, and the other two are phenyl rings. The C1\u2013C10 naphthyl unit is more planar then C11\u2013C20 naphthyl one in which atoms C12, C16, C17, C18 and C19 deviate by 0.051\u2005(3), 0.070\u2005(3), 0.059\u2005(3), 0.082\u2005(3) and 0.054\u2005(3)\u2005\u00c5, respectively, from the mean plane. The two naphthyl moieties are inclined to one another by 72.08\u2005(5)\u00b0. The phenyl rings are inclined at 22.26\u2005(14) and 23.86\u2005(13)\u00b0 to the corresponding naphthyl rings. The bond lengths and angles are mostly in good agreement with those observed in the analogous fragments of the gossypol and dianilinegossypol mol\u00adecules (Gdaniec A\u22efO3 (and N2\u2014H2\u22efO7) bond closes a six-membered ring C7\u2014C8\u2014C22\u2014N1\u2014H1A\u22efO3 (and C17\u2014C18\u2014C27\u2014N2\u2014H3\u22efO7), while the other type of hydrogen bond O4\u2014H4\u22efO3 (and O8\u2014H8\u22efO7) forms a five-membered ring C6\u2014C7\u2014O3\u22efH4\u2014O4 (and C16\u2014C17\u2014O7\u22efH8\u2014O8) (Table\u00a01There are two intra\u00admolecular hydrogen bonds in the mol\u00adecule. The N1\u2014H1) Table\u00a01.p-anizidinegossypol mol\u00adecules are incorporated into centrosymmetric dimers typical for gossypol and dianilinogossypol crystal structures by means of a pair of inversion-related hydrogen bonds O5\u2014H5\u22efO3 and a very weak aromatic \u03c0\u2013\u03c0 stacking inter\u00adaction with a Cg\u22efCg distance of 4.182\u2005(2)\u2005\u00c5 where Cg is the centroid of the C31\u2013C36 ring. The packing of these layers in the crystal structure gives rise to wide ragged channels in the [011] direction. The stabilization of the crystal structure is supported by hydro\u00adphobic inter\u00adactions between adjacent layers. The channels in the structure are 5-7\u2005\u00c5 wide and the void volume of each cell is 655\u2005\u00c53, corresponding to 26.6% of the cell volume. Disordered solvated mol\u00adecules, probably solvent and water mol\u00adecules, occupy these voids of the crystal; their contribution to the scattering was removed with the SQUEEZE routine . C-bound H atoms were positioned geometrically and refined using a riding model, with d(C\u2014H) = 0.93\u2005\u00c5 and Uiso = 1.2Ueq(C) for aromatic, d(C\u2014H) = 0.98\u2005\u00c5 and Uiso = 1.2Ueq (C) for methine, d(C\u2014H) = 0.96\u2005\u00c5 and Uiso = 1.5Ueq (C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015020393/hb7516sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015020393/hb7516Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015020393/hb7516Isup3.cmlSupporting information file. DOI: 1433643CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I complex, a discrete binuclear AgI complex, acetate anions, aceto\u00adnitrile solvent mol\u00adecules and water mol\u00adecules. The mol\u00adecular components are linked through O\u2014H\u22efO, N\u2014H\u22efO and O\u2014H\u22efS hydrogen bonds, forming a chain structure along [100].The title compound consists of a mononuclear Ag 2(C3H6N2S)4(C18H15P)2](C2H3O2)2\u00b7[Ag(C2H3O2)(C3H6N2S)(C18H15P)2]2\u00b72C2H3N\u00b74H2O, the AgI ion in the mononuclear neutral complex exhibits a distorted tetra\u00adhedral environment with coordination by two P atoms from tri\u00adphenyl\u00adphosphane (PPh3) ligands, one S atom of an imidazolidine-2-thione (etu) ligand and one O atom of an acetate anion. The binuclear cationic complex comprises two inversion-related [Ag(C3H6N2S)2(C18H15P)] units with AgI ions bridged by two S atoms from etu ligands forming a four-membered Ag\u2013S\u2013Ag\u2013S ring. Each AgI ion is coordinated by a P atom of a PPh3 ligand, two S atoms of bridging etu ligands and the terminal S atom of an etu ligand in a distorted tetra\u00adhedral environment. In the crystal, the mononuclear complex is linked to lattice water mol\u00adecules through O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming a chain along [100]. In addition, the binuclear complex mol\u00adecules are connected to acetate anions and lattice water mol\u00adecules via O\u2014H\u22efO, N\u2014H\u22efO and O\u2014H\u22efS hydrogen bonds, also along [100].In the title compound, [Ag The mol\u00adecule of the binuclear complex lies across a crystallographic inversion center which is at the center of the Ag2S2 core. The two AgI ions are bridged by two S atoms of etu ligands forming a four-membered Ag\u2013S\u2013Ag\u2013S ring. Each AgI ion is in a distorted tetra\u00adhedral coordination geometry formed by a P atom of a PPh3 ligand, the S atoms of two bridging etu ligands and a terminal S atom of an etu ligand.The structures of the mol\u00adecular components of the title compound are shown in Fig.\u00a01A\u22efO6, O7\u2014H7B\u22efO6iv, N6\u2014H6\u22efO4 and N5\u2014H5\u22efO7iii hydrogen bonds was added and the mixture was stirred for 3\u2005h. Ethyl\u00adene\u00adthio\u00adurea was added and the new reaction mixture was heated under reflux for 5\u2005h. The resulting clear solution was filtered off and left to evaporate at room temperature. Crystals suitable for X-ray diffraction, which were deposited upon standing for a week, were filtered off and dried under reduced pressure.Tri\u00adphenyl\u00adphosphane was dissolved in 30\u2005cmUiso(H) = 1.2Ueq(C); 0.99\u2005\u00c5 (CH2) and Uiso(H) = 1.2Ueq(C); 0.98\u2005\u00c5 (CH3) and Uiso(H) = 1.5Ueq(C); 0.88\u2005\u00c5 (NH) and Uiso(H) = 1.2Ueq(N). Water hydrogen atoms were located from a difference Fourier map and were refined with an O\u2014H distance restraint of 0.84\u2005(2)\u2005\u00c5 and Uiso(H) = 1.2Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901600308X/wm5275sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S205698901600308X/wm5275Isup2.hklStructure factors: contains datablock(s) I. DOI: 1452047CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the title compound contains two crystallographically independent mol\u00adecules with the similar conformation, the piperidine rings in both mol\u00adecules adopt a similar distorted chair conformation and have pseudo mirror planes passing through the N\u2014O bond. 22H25F2N4O2, contains two crystallographically independent mol\u00adecules. In one mol\u00adecule, the two benzene rings are oriented at a dihedral angle of 1.93\u2005(10)\u00b0 and in the other mol\u00adecule the corresponding dihedral angle is 7.19\u2005(9)\u00b0. The piperidine rings in the two mol\u00adecules adopt a similar distorted chair conformation, and both have pseudo-mirror planes passing through the N\u2014O bonds. An intra\u00admolecular O\u2014H\u22efN hydrogen bond between the hy\u00addroxy group and the imine N atom is observed in both mol\u00adecules. In the crystal, weak C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds, enclosing R22(6) ring motifs, and weak \u03c0\u2013\u03c0 stacking inter\u00adactions link the mol\u00adecules into a three-dimensional supra\u00admolecular network, with centroid-to-centroid distances between the nearly parallel phenyl and benzene rings of adjacent mol\u00adecules of 3.975\u2005(2) and 3.782\u2005(2)\u2005\u00c5.The asymmetric unit of the title compound, C The six-membered rings (O1/H1/N3/C10/C11/C13) and (O3/H3/N7/C31/C32/C35) are almost planar, and they are oriented at dihedral angles of 0.83\u2005(10) and 0.92\u2005(9)\u00b0, respectively, to the adjacent benzene (B and E) rings.The phenyl [C (N4/C14\u2013C18) and F (N8/C36\u2013C40)] rings are in distorted chair conformations having total puckering amplitudes QT of 0.491\u2005(3)\u2005\u00c5 (for ring C) and 0.509\u2005(3)\u2005\u00c5 (for ring F), and they have pseudo mirror planes passing through the N4\u2014O2 (for ring C) and N8\u2014O4 (for ring F) bonds.The piperidine , with centroid\u2013centroid distances of 3.975\u2005(2) and 3.782\u2005(2)\u2005\u00c5, respectively, may further stabilize the structure.In the crystal, strong intra\u00admolecular O\u2014H\u22efN and weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds Table\u00a02 link the3 (1:1) solution (70\u2005ml) of 5-[diazen\u00adyl]-2-hy\u00addroxy\u00adbenzaldehyde , and heated at 333\u2005K for 2\u2005h. Then, the reaction mixture was left to slowly cool to room temperature. After one day, orange microcrystals were obtained . Orange block-shaped crystals, suitable for X-ray analysis, were obtained by recrystallization from methanol/CHCl3 (1:1) solution by slow evaporation at room temperature after several days (m.p. 473\u2013475\u2005K).The title compound was synthesized by the reaction of 5-[diazen\u00adyl]-2-hy\u00addroxy\u00adbenzaldehyde , 0.97\u2005\u00c5 (for CH2) and 0.98\u2005\u00c5 (for CH), and constrained to ride on their parent atoms, with Uiso(H) = xUeq(C), where x = 1.5 for methyl H atoms and x = 1.2 for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015012049/xu5856sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015012049/xu5856Isup2.hklStructure factors: contains datablock(s) I. DOI: 1408338CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E conformation. An intra\u00admolecular C\u2014H\u22efF hydrogen bond generates an S(6) ring motif. In the crystal, mol\u00adecules are arranged into centrosymmetric dimers via pairs of C\u2014H\u22efF hydrogen bonds. The crystal structure also features C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions. Hirshfeld surface analysis was used to confirm the existence of inter\u00admolecular inter\u00adactions.In the title compund, the enone moiety adopts an 23H14ClFO, the enone moiety adopts an E conformation. The dihedral angle between the benzene and anthracene ring is 63.42\u2005(8)\u00b0 and an intra\u00admolecular C\u2014H\u22efF hydrogen bond generates an S(6) ring motif. In the crystal, mol\u00adecules are arranged into centrosymmetric dimers via pairs of C\u2014H\u22efF hydrogen bonds. The crystal structure also features C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions. Hirshfeld surface analysis was used to confirm the existence of inter\u00admolecular inter\u00adactions.In the title compound, C The anthracene ring system (C10\u2013C23) is twisted at the C9\u2013C10 bond from the (E)-3-(2-chloro-6-fluoro\u00adphen\u00adyl)acryl\u00adaldehyde moiety [maximum deviation = 0.193\u2005(16)\u2005\u00c5 for atom O1] with a C8\u2014C9\u2014C10\u2014C23 torsion angle of \u221261.4\u2005(2)\u00b0. The terminal benzene and anthracene ring systems form a dihedral angle of 63.42\u2005(8)\u00b0. The least-squares plane through the enone moiety [O1/C7\u2013C9) with a maximum deviation of 0.033\u2005(2)\u2005\u00c5 for atom C9] makes dihedral angles of 5.62\u2005(13) and 59.18\u2005(12)\u00b0 with the benzene (C1\u2013C6) and anthracene (C10\u2013C23) rings, respectively. An intra\u00admolecular C8\u2014H8A\u22efF1 hydrogen bond is observed, generating an S(6) ring motif. The bond lengths and angles are comparable with those in previously reported structures inter\u00adactions .In the crystal Fig.\u00a02, the mol1 Table\u00a01 hydrogeng1 Fig.\u00a03 and Cg1\u22efet al., 2012dnorm are shown in Fig.\u00a04The inter\u00admolecular inter\u00adactions of the title compound can be visualized using Hirshfeld surface analysis .The C17\u2014H17ed + id \u223c2.6\u2005\u00c5 on the fingerprint plot. The presence of \u03c0\u2013\u03c0 inter\u00adactions is shown as C\u22efC contacts, which contribute 8.9% of the Hirshfeld surfaces. The presence of these inter\u00adactions can also be shown by the Hirshfeld surfaces mapped by shape index can be also be seen, indicated by the wings of ex Fig.\u00a06 and the ex Fig.\u00a06.A mixture of 9-acetyl\u00adanthracene and 2-chloro-6-fluoro\u00adbenzaldehyde was dissolved in methanol (20\u2005ml). A catalytic amount of NaOH was added to the solution dropwise with vigorous stirring. The reaction mixture was stirred for about 5\u20136\u2005h at room temperature. After stirring, the contents of the flask were poured into ice-cold water (50\u2005ml) and the resulting crude solid was collected by filtration. The compound was dried and purified by repeated recrystallization from acetone solution, forming yellow plates.Uiso(H) = 1.2Ueq(C). The most disagreeable reflections (\u22121\u00a0\u2212\u00a02 4 and \u22121 1 0) were omitted from the final refinement.Crystal data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016005028/hb7569sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016005028/hb7569Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016005028/hb7569Isup3.cmlSupporting information file. DOI: 1470351CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "P21/n, the methyl acrylate and nitro\u00advinyl units are relatively planar with an E conformation about the C=C bonds. The two aromatic rings are inclined to one another by 74.87\u2005(9) and 75.65\u2005(2)\u00b0 for compounds (I) and (II), respectively. In the crystal of (I), chains along the b axis are formed via C\u2014H\u22efO hydrogen bonds. In the crystal of (II), mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, forming sheets parallel to the ac plane.In the title compounds, (I) and (II), both of which crystallize in the monoclinic space group 19H16ClNO5, (I), and C19H15Cl2NO5, (II), both crystallize in the monoclinic space group P21/n. They differ essentially in the orientation of the methyl acetate group, with the C=O bond directed towards the NO2 group in (I) but away from it in (II). In compound (I), the mean plane of the methyl acrylate unit is planar, with a maximum deviation of 0.0044\u2005(2)\u2005\u00c5 for the methyl C atom, while in (II) this deviation is 0.0147\u2005\u00c5. The inter\u00adplanar angles between the two aromatic rings are 74.87\u2005(9) and 75.65\u2005(2)\u00b0 for compounds (I) and (II), respectively. In both compounds, the methyl acrylate and nitro\u00advinyl groups each adopt an E conformation about the C=C bond. In the crystal of (I), mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds forming chains along the b axis. The chains are linked via C\u2014H\u22efCl hydrogen bonds, forming sheets parallel to the ab plane. The sheets are linked via C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional structure. In the crystal of (II), mol\u00adecules are linked by pairs of C\u2014H\u22efO hydrogen bonds, forming inversion dimers with an R22(30) ring motif. The dimers are linked via C\u2014H\u22efO hydrogen bonds, forming sheets parallel to the ac plane and enclosing R44(28) ring motifs. The sheets are linked via parallel slipped \u03c0\u2013\u03c0 inter\u00adactions (inter\u00adcentroid distances are both ca 3.86\u2005\u00c5), forming a three-dimensional structure.The title compounds, C In (I)In both compounds, the nitro\u00advinyl groups [C2=C1\u2014N1], have an b-axis direction n Table\u00a01. The chane Fig.\u00a04. The shee Table\u00a01.ac plane and enclosing via slipped parallel \u03c0\u2013\u03c0 inter\u00adactions, forming a three-dimensional structure, Fig.\u00a06Cg1\u22efCg1i = 3.863\u2005(2)\u2005\u00c5, inter-planar distance = 3.487\u2005(1)\u2005\u00c5, slippage 1.662\u2005\u00c5; Cg1 is the centroid of ring C3\u2013C8; symmetry code: (i) \u2212x\u00a0+\u00a01, \u2212y, z\u00a0+\u00a01, and Cg2\u22efCg2ii = 3.861\u2005(2)\u2005\u00c5, inter-planar distance = 3.506\u2005(2)\u2005\u00c5, slippage = 1.617\u2005\u00c5; Cg2 is the centroid of ring C14\u2013C19; symmetry code: (ii) \u2212x\u00a0+\u00a01, \u2212y, \u2212z\u00a0+\u00a02].In compound (II)f Table\u00a02. The dims Table\u00a02. The sheE)-2-(phen\u00adoxy\u00admeth\u00adyl)-3-phenyl\u00adacrylate gave 12 hits. There is a great variety in the dihedral angle involving the two aromatic rings; from a minimum of ca 47.2\u00b0 in (E)-methyl 2-({2-eth\u00adoxy-6-[(E)-(hy\u00addroxy\u00adimino)\u00admeth\u00adyl]phen\u00adoxy}meth\u00adyl)-3-phenyl\u00adacrylate -2-[(2-nitro\u00adphen\u00adoxy)meth\u00adyl]-3-phenyl\u00adacrylate -3-(2-chloro\u00adphen\u00adyl)-2-{[2-phen\u00adoxy]meth\u00adyl}acrylate (1\u2005mmol) for compound (I)E)-2-{[4-chloro-2-phen\u00adoxy]meth\u00adyl}-3-(2-chloro\u00adphen\u00adyl)acrylate (1\u2005mol) for compound (II)tert-butyl dicarbonate (1.2 equiv) were added and the solutions of the corresponding crude products were stirred at 318\u2013323\u2005K for 2\u2005h, followed by TLC (20% EtOAc and petroleum ether). The solvents were removed under reduced pressure and the residues purified by column chromatography on silica gel to afford pure products. The purified compounds were recrystallized from ethanol, by slow evaporation of the solvent, yielding block-like crystals of compounds (I)The title compounds were prepared in a similar manner using a mixture of methyl (Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016001493/su5265sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989016001493/su5265Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016001493/su5265IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989016001493/su5265Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016001493/su5265IIsup5.cmlSupporting information file. DOI: 1449405, 1449404CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The packing features supra\u00admolecular layers sustained by O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonding.A distorted octa\u00adhedral 2(C12H10N2)3(C6H12NOS2)4]\u00b74C2H3N, comprises a CdII atom, two di\u00adthio\u00adcarbamate (dtc) anions, one and a half trans-1,2-dipyridin-4-yl\u00adethyl\u00adene (bpe) mol\u00adecules and two aceto\u00adnitrile solvent mol\u00adecules. The full binuclear complex is generated by the application of a centre of inversion. The dtc ligands are chelating, one bpe mol\u00adecule coordinates in a monodentate mode while the other is bidentate bridging. The resulting cis-N2S4 coordination geometry is based on an octa\u00adhedron. Supra\u00admolecular layers, sustained by hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) and hy\u00addroxy-O\u2014H\u22efN(bpe) hydrogen bonding, inter\u00adpenetrate to form a three-dimensional architecture; voids in this arrangement are occupied by the aceto\u00adnitrile solvent mol\u00adecules. Additional inter\u00admolecular inter\u00adactions falling within the specified framework have been analysed by Hirshfeld surface analysis, including \u03c0\u2013\u03c0 inter\u00adactions.The asymmetric unit of the title compound, [Cd The di\u00adthio\u00adcarbamate ligands coordinate with significant differences in their Cd\u2014S bond lengths, Table\u00a01d(Cd\u2014Slong) \u2013 d(Cd\u2014Sshort) = 0.15\u2005\u00c5 for the S1-di\u00adthio\u00adcarbamate ligand cf. 0.10\u2005\u00c5 for the S3-ligand. Nevertheless, there is considerable delocalization of \u03c0-electron density in the CdS2C chelate rings as evidenced by the equivalence of the associated C\u2014S bond lengths, Table\u00a01trans [178.06\u2005(3)\u00b0] and the less tightly bound sulfur atoms are trans to nitro\u00adgen atoms, Table\u00a01cis. The distortions from the ideal geometry are readily related to the restricted bite angles of the chelating ligands, Table\u00a01i and C22\u2014C21\u2014C24\u2014C25 torsion angles of \u221212.2\u2005(6) and 13.9\u2005(5)\u00b0 for the bi- and mono-dentate ligands, respectively; symmetry code: (i) 2\u00a0\u2212\u00a0x, \u2212y, 1\u00a0\u2212\u00a0z.The mol\u00adecular structure of the binuclear title compound, {Cdn TONTO inter\u00adactions and pyridyl-C\u2014H\u22efN(acetonitrile) inter\u00adactions. The presence of short inter\u00admolecular C\u22efC and C\u22efH contacts, Table\u00a03A atoms in Fig.\u00a03i link of the bridging bpe ligand can be viewed as a bright-red region around the C18 atom in the dnorm mapped surface, Fig.\u00a03b; this arises as it is the asymmetric unit that has been investigated not the entire binuclear mol\u00adecule. With respect to the aceto\u00adnitrile molecule the dnorm mapped surfaces show only the aceto\u00adnitrile-N7 to be involved in a significant inter\u00admolecular C\u2014H\u22efN inter\u00adaction \u2005\u00c5, symmetry code: 3\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 2\u00a0\u2212\u00a0z] is reflected through the appearance of green points around de = di \u223c1.8\u2005\u00c5, the red and blue triangle pairs on the Hirshfeld surface mapped with shape-index property identified with arrows in the image of Fig.\u00a07g, with the tips at de + di \u223c2.95\u2005\u00c5 arising from the short inter\u00adatomic S\u22efH/H\u22efS contacts listed in Table\u00a03In the absence of C\u2014H\u22ef\u03c0 inter\u00adactions in the crystal, the pair of characteristic wings resulting in the fingerprint plot delineated into C\u22efH/H\u22efC contacts with 15.9% contribution to the Hirshfeld surface, Fig.\u00a06et al., 2014i.e. 0.97, is in accord with expectation , may be calculated on the basis of Hirshfeld surface analysis 2(\u03bc-bpe)]n, which is a linear coordination polymer with a trans-N2S4 donor set zinc atoms 2]2(\u03bc-bpe) species still forms with non-coordinating bpe included in the structure 2]2(\u03bc-bpe) and one-dimensional [Zn(S2COEt)2(\u03bc-bpe)]n are formed with the dimensionality correlated with the steric bulk of the xanthate-bound R groups 2]2(\u03bc-bpe)}n, R = iPr and Cy (\u03bc2-NO3)(\u03bc-bpe)(bpe)(OH2)]n 2(\u03bc-bpe)(bpe)2(OH2)4]n (\u03bc-bpe)(bpe)2(OH2)2]NO3(bpe)(H2O)4.45 i.e. 2:1, 1:1 or 1:2, between the precursor mol\u00adecules. In a typical experiment, Cd[S2CN(iPr)CH2CH2OH]2 was dissolved in boiling aceto\u00adnitrile (30\u2005ml). trans-1,2-Dipyridin-4-yl\u00adethyl\u00adene was added to this solution, which was allowed to slowly cool to room temperature. Yellow prisms precipitated within an hour. The yield was not measured but was close to qu\u00adanti\u00adtative based on Cd. M.p. = 463\u2013465\u2005K (uncorrected). IR : 1607 m, 1449 ms, 1407 s, 1170 s, 1037 s, 968 s, 954 s, 824 s. NMR: 1H \u03b4 (p.p.m.): 8.6 , 7.6 , 7.54 , 5.21 , 4.82 , 3.76-3.67 , 1.18 . TGA: one sharp step followed by a protracted mass loss totalling 71.5%, assigned to decomposition to CdS .The title compound was isolated regardless of the ratio, Uiso(H) set to 1.2\u20131.5Ueq(C). The oxygen-bound H atoms were located in a difference Fourier map but were refined with a distance restraint of O\u2014H = 0.84\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989016010768/hb7594sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016010768/hb7594Isup2.hklStructure factors: contains datablock(s) I. DOI: 1489732CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the two independent mol\u00adecules stack head-to-tail probably due to the presence of the dipole moment of the 11H12N6O2, a \u03c0-conjugated triazene, crystallized with two independent mol\u00adecules (A and B) in the asymmetric unit. Both mol\u00adecules have an E conformation about the \u2013N=N\u2013 bond and have slightly twisted overall conformations. In mol\u00adecule A, the imidazole ring is inclined to the benzene ring by 8.12\u2005(4)\u00b0, while in mol\u00adecule B the two rings are inclined to one another by 7.73\u2005(4)\u00b0. In the crystal, the independent mol\u00adecules are linked to each other by C\u2014H\u22efO hydrogen bonds, forming \u2013A\u2013A\u2013A\u2013 and \u2013B\u2013B\u2013B\u2013 chains along [100]. The chains are linked by C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds, forming sheets lying parallel to (001). The sheets are linked by further C\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions , forming a three-dimensional framework structure.The title compound, C The structure of the triazene moiety is influenced by the resonance arising from delocalization of the electron lone-pair on the third N atom, towards the double bond. Triazenes are relatively old compounds from the organic chemist\u2019s viewpoint. It was as early as 1862 that Griess described a suitable method for the synthesis of 1,3-di\u00adphenyl\u00adtriazene of the title compound are illustrated in Fig.\u00a01E conformation about the \u2013N5=N4\u2013 and \u2013N11=N10\u2013 bonds and the bond lengths and angles of the \u03c0-conjugated triazene unit are inclined to one another by 7.73\u2005(4)\u00b0.The mol\u00adecular structures of the two independent mol\u00adecules ; see Fig.\u00a02Cg1\u22efCg4i = 3.5243\u2005(5)\u2005\u00c5; Cg1 and Cg4 are the centroids of the imidazole ring of mol\u00adecule A and the benzene ring of mol\u00adecule B; symmetry code: (i) x, y, z\u00a0\u2212\u00a01], forming a three-dimensional framework structure : \u03b4 7.99 , 7.85\u20137.83 , 7.70\u20137.69 , 7.55\u20137.52 , 7.06 3.60 . 13C NMR : \u03b4 154.4, 151.1, 149.0, 130.6, 126.9, 118.8, 118.3, 114.4, 35.7. UV/Vis : \u03bb (\u220a) = 455\u2005nm. HRMS : m/z calculated for C11H13N6O2 [M + H]+ 261.1095, found 261.1094.1-Azido-3-nitro\u00adbenzene was prepared according to the literature procedure = 1.5Ueq(C) for methyl H atoms and = 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S1600536814020698/su2778sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814020698/su2778Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814020698/su2778Isup3.cmlSupporting information file. DOI: 977732CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Supra\u00admolecular layers sustained by N\u2014H\u22efN and N\u2014H\u22ef\u03c0 inter\u00adactions are found in the crystal packing; these are connected by \u03c0\u2013\u03c0 contacts.A penta\u00adcoordinated Zn 32H16N8)(C7H9N)]\u00b73C7H9N, comprises two independent complex mol\u00adecules and three benzyl\u00adamine solvent mol\u00adecules. Each complex mol\u00adecule features a penta\u00adcoordinated Zn2+ ion within a square-pyramidal geometry, whereby the N5 donor set is defined by four atoms of the phthalocyaninate dianion (PC) and an N-bound benzyl\u00adamine mol\u00adecule; it is the relative orientations of the latter that differentiate between the independent complex mol\u00adecules. The uncoordinated benzyl\u00adamine mol\u00adecules display different conformations in the structure, with syn-Car\u2014Car\u2014Cm\u2014N torsion angles spanning the range \u221228.7\u2005(10) to 35.1\u2005(14)\u00b0. In the crystal, N\u2014H\u22efN and N\u2014H\u22ef\u03c0 inter\u00adactions lead to supra\u00admolecular layers in the ab plane. The layers have a zigzag topology, have the coordinating and non-coordinating benzyl\u00adamine mol\u00adecules directed to the inside, and present the essentially flat PC resides to the outside. This arrangement enables adjacent layers to associate via \u03c0\u2013\u03c0 inter\u00adactions [inter-centroid distance between pyrrolyl and fused-benzene rings = 3.593\u2005(2)\u2005\u00c5] so that a three-dimensional architecture is formed.The asymmetric unit of the title compound, 2[Zn(C Recent inter\u00adest has centered on their electronic, photoelectronic and catalytic properties for a diverse array of applications including photodynamic therapy is still less than 7\u2005mM Zinc phthalocyanine (ZnPC) is one of the more soluble members of the transition metal phthalocyanines, although a saturated solution in NMP (i.e. 2.025\u2005(3)\u2005\u00c5 [Zn1\u2014N6] to 2.045\u2005(3) [Zn1\u2014N4] \u00c5, suggesting extensive delocalization of \u03c0-electron density over the PC chromophore. Further, the Zn\u2014N(PC) bond lengths are systematically shorter than the Zn\u2014N(amino) bonds. The N5 donor set defines an approximately square-pyramidal geometry with the benzyl\u00adamino-N atoms occupying the axial position. In this description, Zn1 lies 0.4670\u2005(16)\u2005\u00c5 above the least-squares plane defined by the four PC-N atoms (r.m.s. deviation = 0.0104\u2005\u00c5) in the direction of the benzyl\u00adamino-N atom [2.570\u2005(4)\u2005\u00c5 above the plane]; the comparable values for the Zn2-containing mol\u00adecule are 0.4365\u2005(16), 0.0076 and 2.549\u2005(4)\u2005\u00c5, respectively. That the N5 donor set defines a square pyramid is qu\u00adanti\u00adfied by the value of \u03c4 = 0.02 for each of the Zn1- and Zn2-containing mol\u00adecules, which compares to the \u03c4 values of 0.0 and 1.0 for ideal square-pyramidal and trigonal\u2013bipyramidal geometries, respectively angles collated in Table\u00a01The asymmetric unit of (I)As seen from the overlay diagram in Fig.\u00a02Database survey.A discussion of the uncoordinated benzyl\u00adamine mol\u00adecules is found below in the PLATON inter\u00adaction. The second amine-H atom of the coordinating benzyl\u00adamine mol\u00adecule forms an N\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adaction with a non-coordinating N21-benzyl\u00adamine mol\u00adecule. For the Zn2-containing mol\u00adecule, the coordinating N18-benzyl\u00adamine forms a donor N\u2014H\u22efN hydrogen bond to a non-coordinating N20-benzyl\u00adamine mol\u00adecule which is folded to enable a donor N\u2014H\u22efN hydrogen bond to an exocyclic-N atom of the PC dianion. As seen from the lower view of Fig.\u00a03ab plane. These have a zigzag topology and present the flat PC residues to the outside with the benzyl\u00adamine mol\u00adecules, both coordinating and non-coordinating, in the inter-layer region. The layers stack along the c axis being connected by \u03c0\u2013\u03c0 inter\u00adactions between pyrrolyl and fused-phenyl rings [inter-centroid \u22ef(C57\u2013C62) distance = 3.593\u2005(2)\u2005\u00c5 with an angle of inclination = 6.1\u2005(2)\u00b0]. A view of the unit cell contents is shown in Fig.\u00a04Based on the standard criteria incorporated within i.e. with 4-methyl\u00adpridine 2L2 , syn and anti-C(phen\u00adyl)\u2014C(phen\u00adyl)\u2014C(benz\u00adyl)\u2014N(amine) torsion-angle data collated in Table\u00a04et al., 2010Finally, a few comments on the benzyl\u00adamine mol\u00adecule which has now been characterized in its uncoordinated form in five crystal structures. From the et al., 2007\u22121) \u03bd 1608 s (C=N), 1584 w (C=C), 1377 m, 1334 m, 1285 m, 1164 w, 1118 m, 1088 m, 1060 m, 888 w, 878 w, 752 m, 728 s. Crystals of (I)Zinc phthalocyanine was prepared by a modification of a literature procedure (Bayo Uiso(H) set to 1.2Ueq(C). The N-bound H atoms were treated similarly with N\u2014H = 0.88\u2005\u00c5, and with Uiso(H) = 1.2Ueq(N). Each of three solvent benzyl\u00adamine mol\u00adecules suffered from high thermal motion. In the final refinement the benzene rings were constrained to be regular hexa\u00adgons (C\u2014C = 1.39\u2005\u00c5) and their ADP\u2019s restrained to be nearly isotropic using the ISOR command. Owing to poor agreement, two reflections, i.e. (0 3 2) and (4 0 14), were omitted from the final cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989015014280/hb7468sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015014280/hb7468Isup2.hklStructure factors: contains datablock(s) I. DOI: 1415614CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Weak C\u2014H\u22efCl inter\u00admolecular inter\u00adactions result in a two-dimensional extended sheet structure normal to the b axis.The mol\u00adecular structure of [Fe 4(C5H7O2)4(CH3O)6Cl2] or [Fe4(acac)4(\u03bc2-OMe)4(\u03bc3-OMe)2Cl2] (acac = acetyl\u00adacetonate), crystallizes in the ortho\u00adrhom\u00adbic Pbca space group with one half of the mol\u00adecule per asymmetric unit, the other half being completed by inversion symmetry. The core structure consists of a face-sharing double pseudo-cubane entity with two opposite corners missing. Weak C\u2014H\u22efCl inter\u00admolecular inter\u00adactions result in a two-dimensional layered structure parallel to the ac plane.The title complex, [Fe In line with this observation, we now report that the addition of NaOSi(OMe)2Me to Fe(acac)2Cl results in the formation of a tetra\u00adnuclear iron(III) methano\u00adlate compound, Fe4(acac)4(\u03bc2-OMe)4(\u03bc3-OMe)2Cl2, (I)We have investigated the syntheses of metal meth\u00adoxy\u00adsilanolates III metal atoms. Both cations are in approximately octa\u00adhedral coordination environments. The coordination sphere of Fe1 is filled by the O atoms of one \u03ba2-acac ligand [Fe1\u2014O1 = 1.9971\u2005(13)\u2005\u00c5 and Fe1\u2014O2 = 1.9934\u2005(13)\u2005\u00c5], two \u03bc2-methano\u00adlate groups , one \u03bc3-methano\u00adlate group [Fe1\u2014O4 = 2.2135\u2005(12)\u2005\u00c5], and one terminal chloride ligand [Fe1\u2014Cl1 = 2.2776\u2005(5)\u2005\u00c5]. The coordination sphere of Fe2 is filled by the O atoms of one \u03ba2-acac ligand [Fe2\u2014O6 = 1.9717\u2005(13)\u2005\u00c5 and Fe2\u2014O7 = 1.9692\u2005(12)\u2005\u00c5], two \u03bc2-methano\u00adlate groups [Fe2\u2014O3 = 1.9755\u2005(12)\u2005\u00c5 and Fe2\u2014O5 = 1.9823\u2005(12)\u2005\u00c5], and two \u03bc3-methano\u00adlate groups [Fe2\u2014O4 = 2.0815\u2005(12)\u2005\u00c5 and Fe2\u2014O4i = 2.0809\u2005(12)\u2005\u00c5]. The angles around both Fe1 and Fe2 distort significantly from the ideal values of 90 and 180\u00b0 of a perfect octa\u00adhedron. For Fe1, the cis angles range from 75.69\u2005(5) to 98.40\u2005(4)\u00b0, while the trans angles range from 164.47\u2005(5) to 170.40\u2005(3)\u00b0. The angles around Fe2 have narrower ranges, with cis being 78.95\u2005(5)\u201396.48\u2005(5)\u00b0 and trans being 170.08\u2005(5)\u2013170.16\u2005(5)\u00b0.The structure of (I)4(OMe)6] face-sharing double pseudo-cubane entity with two opposite corners missing. The outside of the cluster is decorated by one acac ligand per metal and the Fe atoms at either end of the cluster are coordinated by one chloride ion. Neighboring Fe\u22efFe distances range from 3.1997\u2005(4) to 3.2175\u2005(6)\u2005\u00c5, while the Fe1\u22efFe1i distance is 5.5702\u2005(6)\u2005\u00c5.The mol\u00adecular structure of (I)\u2212 ion and an acac ligand on neighboring mol\u00adecules s Table\u00a01. Taking is Fig.\u00a02.4(acac)4(OMe)6(N3)2], has previously been reported 4(OMe)6] face-sharing double cubane cluster with two opposite corners missing. The average Fe\u2014Oacac distance of 1.978\u2005\u00c5 is quite close to the average Fe\u2014Oacac distance of 1.982\u2005\u00c5 in (I)2-OMe: 1.977\u2005\u00c5; \u03bc3-OMe: 2.124\u2005\u00c5) are also comparable to those in (I)2-OMe: 1.983\u2005\u00c5; \u03bc3-OMe: 2.125\u2005\u00c5).One closely related complex, [Fe4(OR)6] cluster core similar to (I)et al., 2009et al., 20094(OR)6] motif is present is 63 additional materials as part of a higher-order cluster complex in THF (3\u2005ml) was added to a solution of Fe(acac)2Cl in THF (see Scheme). The mixture was stirred rapidly at room temperature, and a slight color change from a dark-red to a lighter red was observed. Removal of the solvent under vacuum resulted in the precipitation of an orange solid, which upon washing with dry Et2O (2 \u00d7 10\u2005ml) left a yellow solid. The yellow solid was extracted into dry CH2Cl2 and filtered through Celite. The CH2Cl\u00ad2 was then removed under vacuum, leaving a yellow solid . Crystals suitable for X-ray diffraction were grown by slow diffusion of pentane into a CH2Cl2 solution of the yellow solid.A solution of NaOSi(OMe)RCH3, were optimized by rotation about R\u2014C bonds, with idealized C\u2014H, R\u2014H and H\u22efH distances (C\u2014H = 0.98\u2005\u00c5). The remaining H atoms were included as riding idealized contributors (C\u2014H = 0.95\u2005\u00c5). H atoms were assigned Uiso(H) = 1.5Ueq(C) for methyl H atoms and Uiso(H) = 1.2Ueq(C) otherwise. The 102 reflection was omitted from the final refinement because it was partially obscured by the shadow of the beam stop.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015013535/cv5491sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015013535/cv5491Isup2.hklStructure factors: contains datablock(s) I. DOI: 1412851CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atom in the title compound shows a slightly distorted octa\u00adhedral coordination environment to four N atoms of the aza\u00admacrocylic ligand in the equatorial plane and two isonicotinate O atoms in axial positions. Inter\u00admolecular N\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions consolidate the crystal packing.The Ni 6H4NO2)2(C16H38N6)], was prepared through self-assembly of a nickel(II) aza\u00admacrocyclic complex with isonicotinic acid. The NiII atom is located on an inversion center and exhibits a distorted octa\u00adhedral N4O2 coordination environment, with the four secondary N atoms of the aza\u00admacrocyclic ligand in the equatorial plane [average Ni\u2014Neq = 2.064\u2005(11)\u2005\u00c5] and two O atoms of monodentate isonicotinate anions in axial positions [Ni\u2014Oax = 2.137\u2005(1)\u2005\u00c5]. Intra\u00admolecular N\u2014H\u22efO hydrogen bonds between one of the secondary amine N atoms of the aza\u00admacrocyclic ligand and the non-coordinating carboxyl\u00adate O atom of the anion stabilize the mol\u00adecular structure. Inter\u00admolecular N\u2014H\u22efN hydrogen bonds, as well as \u03c0\u2013\u03c0 inter\u00adactions between neighbouring pyridine rings, give rise to the formations of supra\u00admolecular ribbons extending parallel to [001].The title compound, [Ni(C The longer axial bonds can be attributed to a ring contraction of the aza\u00admacrocyclic ligand adopts the expected chair conformation, whereas the five-membered NiC2N2 ring (Ni1\u2013N1\u2013C1\u2013C4\u2013N2) has a gauche conformation (ClO4)2], was prepared in a slightly modified procedure with respect to the reported method (ClO4)2] was slowly added an aceto\u00adnitrile solution (8\u2005mL) containing isonicotinic acid and excess tri\u00adethyl\u00adamine at room temperature. The purple precipitate was filtered off, washed with aceto\u00adnitrile and diethyl ether, and dried in air. Single crystals of compound (l)16H38N6)(ClO4)2] for several days. Yield: 0.167\u2005g (52%). FT\u2013IR : 3145, 3075, 2951, 2920, 1571, 1457, 1351, 1272, 1014, 915.The starting complex, [Ni values of 1.2 or 1.5Ueq of the parent atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016001031/wm5263sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016001031/wm5263Isup2.hklStructure factors: contains datablock(s) I. DOI: 1447865CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the cations, anions, 4-hy\u00addroxy-3-meth\u00adoxy\u00adbenzaldehyde and water mol\u00adecules of crystallization are linked by classical O\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds into a three-dimensional supra\u00admolecular architecture. The crystal packing is further stabilized by weak C\u2014H\u22ef\u03c0 inter\u00adactions involving pyrrole and benzene rings. \u03c0\u2013\u03c0 stacking between parallel benzene rings of adjacent 4-hy\u00addroxy-3-meth\u00adoxy\u00adbenzaldehyde mol\u00adecules is also observed, the centroid\u2013centroid distance being 3.8003\u2005(13)\u2005\u00c5. The three F atoms of the anion are disordered over two sets of sites, with a refined occupancy ratio 0.527\u2005(12):0.473\u2005(12). The O atom of one water mol\u00adecule of crystallization is also disordered over two positions in an occupancy ratio of 0.68\u2005(5):0.32\u2005(5).In the title compound, [Fe(C DOI: 10.1107/S1600536814015335/xu5800Isup2.hklStructure factors: contains datablock(s) I. DOI: 975656CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, weak N\u2014H\u22efS hydrogen bonds between the amino groups and the thio\u00adcyanate anions form an R42(8) motif. The complex mol\u00adecules are linked by O\u2014H\u22efO, O\u2014H\u22efS, and N\u2014H\u22efS hydrogen bonds into a three-dimensional supra\u00admolecular structure. Weak \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings is also found [centroid\u2013centroid distance = 3.8578\u2005(14)\u2005\u00c5].In the title complex, [Ni(NCS) DOI: 10.1107/S1600536814006771/hy2643Isup2.hklStructure factors: contains datablock(s) I. DOI: 993930CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked via pairs of N\u2014H\u22efO hydrogen bonds, forming inversion dimers which are linked by O\u2014H\u22efO hydrogen bonds, involving the crystal water mol\u00adecule, forming chains propagating along the a-axis direction.The title compound is twisted in such a way that the almost planar segments are inclined to on another by 77.36\u2005(8)\u00b0, while the benzene rings are normal to one another, making a dihedral angle of 89.69\u2005(9)\u00b0. In the crystal, the water mol\u00adecule links three mol\u00adecules through two O\u2014H\u22efO and one N\u2014H\u22efO hydrogen bonds. The mol\u00adecules are linked via pairs of N\u2014H\u22efO hydrogen bonds, forming inversion dimers with an R22(14) ring motif. The dimers are linked by O\u2014H\u22efO hydrogen bonds, involving two mol\u00adecules of water, forming chains along [100], enclosing R22(14) and R22(18) ring motifs. The chains are linked through C\u2014H\u22efO inter\u00adactions, forming sheets parallel to (010). Within the sheets, there are C\u2014H\u22ef\u03c0 and parallel slipped \u03c0\u2013\u03c0 stacking inter\u00adactions present [inter-centroid distance = 3.6458\u2005(12)\u2005\u00c5].The title compound, C N-Acyl\u00adhydrazones have been reported to be promising in terms of their future potential as anti\u00adbacterial drugs and the C\u2014H(imine) bonds. The C9\u2014O2 bond length of 1.2251\u2005(19)\u2005\u00c5 indicates that the mol\u00adecule exists in the keto form in the solid state, and the C10\u2014N3 bond length of 1.271\u2005(2)\u2005\u00c5 confirms its significant double-bond character. The C9\u2014N2 and N2\u2014N3 bond distances of 1.351\u2005(2) and 1.3771\u2005(18)\u2005\u00c5, respectively, indicate a significant delocalization of the \u03c0-electron density over the hydrazone portion of the mol\u00adecule. Variations in the C\u2014N bond lengths of 1.330\u2005(2), 1.442\u2005(2) and 1.351\u2005(2)\u2005\u00c5 for C7\u2014N1, C8\u2014N1 and C9\u2014N2, respectively, characterize mobility of the bridge and the integral flexibility of the \u2013C(=O)\u2013NH\u2013CH2C(=O)\u2013NH\u2013N=CH\u2013 group connecting the two benzene rings. The mol\u00adecule is twisted at atom C8, the C7\u2014N1\u2014C8\u2014C9 torsion angle being 79.8\u2005(2)\u00b0. The hydrazone part of the mol\u00adecule is almost planar, with C9\u2014N2\u2014N3\u2014C10 and N2\u2014N3\u2014C10\u2014C11 torsion angles of \u2212177.07\u2005(15) and 179.38\u2005(14)\u00b0, respectively. Further, the dihedral angle between the almost planar hydrazone segment \u2005\u00c5 for atom N2) and the attached benzene ring (C11\u2013C16) is 8.17\u2005(6)\u00b0. The two benzene rings (C1\u2013C6 and C11\u2013C16) are orthogonal to each other, making a dihedral angle of 89.69\u2005(9)\u00b0. The planar amide segment is inclined to the attached toluene ring (C1\u2013C6) by 8.06\u2005(9)\u2005\u00c5.The title compound crystallizes as a monohydrate Fig.\u00a01. The conN\u22efO3). A pair of N2\u2014H2N\u22efO1 inter\u00admolecular hydrogen bonds link the mol\u00adecules, forming inversion dimers, with an via hydrogen bonds involving the water mol\u00adecule generating N\u22efO1 and N1\u2014H1N\u22efO3 hydrogen bonds between the mol\u00adecules of the main compound and water mol\u00adecules translate into a-axis direction (Table\u00a01Cg2\u22efCg2i = 3.6458\u2005(12)\u2005\u00c5; inter-planar distance = 3.4135\u2005(8)\u2005\u00c5, slippage = 1.281\u2005\u00c5; Cg2 is the centroid of ring C11\u2013C16; symmetry code: (i) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01] involving inversion-related chloro\u00adbenzene rings; see Fig.\u00a03In the crystal of (I)n Table\u00a01 The chai2\u2013C(=O)\u2013NH\u2013N=CH\u2013, yielded only one hit, namely N-(2-hy\u00addroxy-1-naphthyl\u00admethyl\u00adene)-N\u2032-(N-phenyl\u00adglyc\u00adyl)hydrazine -N-{2-[2-(3-chlorobenzyl\u00adidene)hydrazin\u00adyl]-2-oxoeth\u00adyl}-4-methyl\u00adbenzene\u00adsulf\u00adonamide monohydrate (II) N1) and hydrazone segment (N2\u2013N3=C10\u2013C11) with respect to the glycinyl C8\u2014C9 bond in (I)cis orientation of the sulfonamide and hydrazone segments, with respect to the glycinyl C\u2014C bond, observed in compound (II). In the structure of (I)A search of the Cambridge Structural Database was added slowly. The organic phase was separated and washed with water (30\u2005ml), dried with anhydrous Na2SO4 and evaporated to yield the corresponding ester, N-(4-methyl\u00adbenzo\u00adyl)glycine ethyl ester (L1). L1 (0.01\u2005mol) was added in small portions to a stirred solution of 99% hydrazine hydrate (10\u2005ml) in 30\u2005ml ethanol. The mixture was refluxed for 6\u2005h. After cooling to room temperature, the resulting precipitate was filtered, washed with cold water and dried to give N-(4-methyl\u00adbenzo\u00adyl)-glycinyl hydrazide (L2). 2-Chloro\u00adbenzaldehyde (0.01\u2005mol) and two drops of glacial acetic acid were added to L2 (0.01\u2005mol) in anhydrous methanol (30\u2005ml). The reaction mixture was refluxed for 8\u2005h. After cooling, the precipitate was collected by vacuum filtration, washed with cold methanol and dried. It was recrystallized to constant melting point from methanol (479\u2013480\u2005K). Prism-like colourless single crystals of the title compound were grown from a solution in DMF by slow evaporation of the solvent.Tri\u00adethyl\u00adamine (0.03\u2005mol) and 4-methyl\u00adbenzoyl chloride (0.01\u2005mol) were added to a stirred suspension of glycine ethyl\u00adester hydro\u00adchloride (0.01\u2005mol) in di\u00adchloro\u00admethane (50\u2005ml) in an ice bath. The reaction mixture was stirred at room temperature for 20\u2005h. After completion of the reaction, 2\u22121 for the stretching bands of N\u2014H (amide I), N\u2014H (amide II), C=O(hydrazone), C=O(amide) and C=N, respectively. The characteristic 1H and 13C NMR spectra of the title compound are as follows: 1H NMR : 2.36 , 4.01, 4.45 , 7.25 , 7.33\u20137.40 , 7.42\u20137.45 , 7.81 , 7.97\u20137.99 , 8.39, 8.63 , 8.54, 8.76 , 11.65, 11.73 . 13C NMR : 20.97, 40.74, 42.04, 126.60, 126.83, 127.28, 128.64, 129.66, 130.85, 131.35, 133.10, 139.45, 141.06, 142.70, 165.98, 166.54, 170.48.The purity of the compound was checked by TLC and characterized by its IR spectrum. The characteristic absorptions observed are 3323.3, 3203.8, 1685.8, 1620.2 and 1566.2\u2005cmUiso(H) = 1.5Ueq(O) and 1.2Ueq(N). The C-bound H atoms were positioned with idealized geometry and refined as riding atoms: C\u2014H = 0.93\u20130.97\u2005\u00c5 with Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015011147/su5148sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015011147/su5148Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015011147/su5148Isup3.cmlSupporting information file. DOI: 1405614CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(6) graph-set motif. In the crystal, pairs of C\u2014H\u22efCl hydrogen bonds link mol\u00adecules into inversion dimers with an R22(26) motif. Weak C\u2014H\u22efO inter\u00adactions further link these dimers into ribbons propagating in [100].The hy\u00addroxy group in this carbazole derivative is involved in an intra\u00admolecular O\u2014H\u22efO hydrogen bond, which generates an 25H15Cl2NO4S, the di\u00adchloro\u00adphenyl ring is twisted by 68.69\u2005(11)\u00b0 from the mean plane of the carbazole ring system [r.m.s. deviation = 0.084\u2005(2)\u00b0]. The hy\u00addroxy group is involved in an intra\u00admolecular O\u2014H\u22efO hydrogen bond, which generates an S(6) graph-set motif. In the crystal, pairs of C\u2014H\u22efCl hydrogen bonds link mol\u00adecules into inversion dimers with an R22(26) motif. Weak C\u2014H\u22efO inter\u00adactions further link these dimers into ribbons propagating in [100].In the title compound, C The widening of angle O1\u2014S1\u2014O2 [120.49\u2005(11)\u00b0] and narrowing of angle N1\u2014S1\u2014C1 [105.36\u2005(10)\u00b0] from the ideal tetra\u00adhedral values are attributed to the Thorpe\u2013Ingold effect graph-set motif. In the crystal, pairs of C\u2014H\u22efCl hydrogen bonds link mol\u00adecules into inversion dimers with an et al., 1995The hy\u00addroxy group is involved in an intra\u00admolecular O\u2014H\u22efO hydrogen bond Table\u00a01, which g2 in dry DMF (20\u2005ml) at reflux for 1\u2005h under N2. The reaction mass was poured over crushed ice (50\u2005ml) containing concentrated HCl (1\u2005ml). The precipitated solid was filtered, washed with water and air-dried to obtain the crude compound, which was purified by flash column chromatography on silica gel to afford 17\u2005g as pale-yellow crystals suitable for X-ray analysis. Yield: 368\u2005mg (78%); m.p.: 461\u2013463\u2005K.Enamine 16\u2005g was reacted with CuBrUiso(H) = 1.2Ueq(C). The components of the anisotropic displacement parameters in the direction of the bond between O4 and C19 were restrained to be equal within an effective standard deviation of 0.001 using the DELU command in SHELXL97 global, I. DOI: 10.1107/S1600536814024064/cv5474Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814024064/cv5474Isup3.cmlSupporting information file. DOI: 1032055CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II complex coordinated by hepta\u00adfluoro\u00adbutanoic acid, 1,10-phenanthroline and a water mol\u00adecule is described.The mol\u00adecular structure of a mononuclear Cu 4F7O2)2(C12H8N2)(H2O)], is mononuclear and contains a penta\u00adcoordinated CuII ion. The geometry of CuII ion can be described as distorted square-pyramidal with two O atoms of two butano\u00adate anions and two N atoms of the o-phenanthroline ligand occupying the basal plane, and a water O atom located at the axial position. In the crystal, C\u2014H\u22ef and O\u2014H\u22ef hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid-to-centroid distance 3.533\u2005(2)\u2005\u00c5] link the mol\u00adecules into a three-dimensional supra\u00admolecular structure.The title compound, [Cu(C For perfect tetra\u00adgonal symmetry, \u03c4 is zero, and for perfect trigonal\u2013bipyramidal geometry, \u03c4 becomes 1.0 and 1.980\u2005(3)\u2005\u00c5] in the quadrilateral plane are shorter than the apical position [2.173\u2005(3)\u2005\u00c5]. The mean Cu\u2014N(phen) distance of 2.043\u2005\u00c5 and the bite angle N1\u2014Cu1\u2014N2 of 81.75\u2005(12)\u00b0 are close to the corresponding values observed in related copper\u2013via atoms H5A and H5B, to oxygen atom O3 of one coordinating carboxyl\u00adate group (\u2212x\u00a0+\u00a0y\u00a0+\u00a0z) and to the dangling oxygen atom O2 of the other coordinating carboxyl\u00adate group (\u2212x\u00a0+\u00a0y\u00a0+\u00a0z), thus enclosing centrosymmetric et al., 1995b-axis direction \u2005\u00c5. was added to a solution of o-phenanthroline and hepta\u00adfluoro\u00adbutanoic acid in methanol (7\u2005ml). Afterwards the obtained transparent blue solution was left to evaporate slowly in the air at ambient temperature and after two weeks, X-ray quality crystals appeared as blue plates. They were filtered off, washed with diethyl ether and dried in the air. Yield: 46\u2005mg, 86%.Cu(ClO4)\u00b76HUiso(H) = 1.2Ueq(C). The coordinates of the water H atoms were refined, and Uiso(H) was set to be 2Ueq(O). One of the hepta\u00adfluoro\u00adbutano\u00adate groups is disordered over two sets of sites in a 0.705\u2005(9):0.955\u2005(9) ratio. Atoms associated with the disorder were refined with isotropic displacement parameters.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015022720/pj2026sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015022720/pj2026Isup2.hklStructure factors: contains datablock(s) I. DOI: 1434356CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The hydrogen-bonded structures of both a (1:1) mol\u00adecular adduct and a salt of 5-(4-bromo\u00adphen\u00adyl)-1,3,4-thia\u00addiazol-2-amine with 4-nitro\u00adbenzoic acid and 3,5-di\u00adnitro\u00adsalicylic acid, respectively, have been determined. 8H6BrN3S\u00b7C7H5NO4, (I), and the salt C8H7BrN3S+\u00b7C7H3N2O7\u2212, (II), obtained from the inter\u00adaction of 5-(4-bromo\u00adphen\u00adyl)-1,3,4-thia\u00addiazol-2-amine with 4-nitro\u00adbenzoic acid and 3,5-di\u00adnitro\u00adsalicylic acid, respectively, have been determined. The primary inter-species association in both (I) and (II) is through duplex R22(8) (N\u2014H\u22efO/O\u2014H\u22efO) or (N\u2014H\u22efO/N\u2014H\u22efO) hydrogen bonds, respectively, giving heterodimers. In (II), these are close to planar [the dihedral angles between the thia\u00addiazole ring and the two phenyl rings are 2.1\u2005(3) (intra) and 9.8\u2005(2)\u00b0 (inter)], while in (I) these angles are 22.11\u2005(15) and 26.08\u2005(18)\u00b0, respectively. In the crystal of (I), the heterodimers are extended into a chain along b through an amine N\u2014H\u22efNthia\u00addiazole hydrogen bond but in (II), a centrosymmetric cyclic hetero\u00adtetra\u00admer structure is generated through N\u2014H\u22efO hydrogen bonds to phenol and nitro O-atom acceptors and features, together with the primary R22(8) inter\u00adaction, conjoined R46(12), R21(6) and S(6) ring motifs. Also present in (I) are \u03c0\u2013\u03c0 inter\u00adactions between thia\u00addiazole rings [minimum ring-centroid separation = 3.4624\u2005(16)\u2005\u00c5], as well as short Br\u22efOnitro inter\u00adactions in both (I) and (II) .The structures of the 1:1 co-crystalline adduct C This \u2018planar\u2019 conformation is found in the parent acid \u00adbenzoic acid PNBA adduct with BATZ, (I)ed Fig.\u00a01, in whicanti-related carb\u00adoxy\u00adlic acid H atom forming the common intra\u00admolecular S(6) hydrogen bond which is found in ca. 70% of DNSA salt structures \u00b0] whereas the second nitro group and the carboxyl\u00adate group lie essentially in the plane [torsion angles: C6A\u2014C5A\u2014 N5A\u2014O51A = 179.5\u2005(4) and C2A\u2014C1A\u2014 C11A\u2014O11A = \u2212178.0\u2005(4)\u00b0].In the DNSA salt (II)B\u2014H21B\u22efN4Bi hydrogen bonds s Table\u00a01 forming 1b Fig.\u00a03. This is1b Fig.\u00a03 and the Fig.\u00a03b. A weak B) and the adjacent nitro-O atom (O31A) (Table\u00a02S(6) motifs \u2005\u00c5]. However, unlike in the structure of (I)With (II)) Table\u00a02 gives anfs Fig.\u00a04. The hetnitro contacts are found: for (I)B\u22efO42Aiii = 3.314\u2005(4)\u2005\u00c5, and for (II)B\u22ef O52Aiv = 3.104\u2005(3)\u2005\u00c5 .In both (I)The title compounds were prepared by the reaction of 1\u2005mmol (260\u2005mg) of 5-(4-bromo\u00adphen\u00adyl)-1,3,4-thia\u00addiazol-2-amine with 1\u2005mmol of either 4-nitro\u00adbenzoic acid (167\u2005mg) [for (I)] or 3,5-di\u00adnitro\u00adsalicylic acid (228\u2005mg) [for (II)] in 20\u2005mL of 50% ethanol\u2013water, with 10\u2005min refluxing. Partial evaporation of the solvent gave colourless needles of (I)Uiso(H) = 1.2Ueq(N) or 1.5Ueq(O). Other H atoms were included at calculated positions [C\u2014H = 0.95\u2005\u00c5] and also treated as riding, with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S1600536814021138/lh5731sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S1600536814021138/lh5731Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S1600536814021138/lh5731IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S1600536814021138/lh5731Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S1600536814021138/lh5731IIsup5.cmlSupporting information file. DOI: 1025540, 1025541CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N6-benzoyl\u00adadenine-adipic acid (1/0.5) is reported. The typical C=O\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions are also present in this structure.The supra\u00admolecular architecture in a co-crystal of N(7)\u2014H tautomeric form of 12H9N5O\u00b70.5C6H10O4, consists of one mol\u00adecule of N6-benzoyl\u00adadenine (BA) and one half-mol\u00adecule of adipic acid (AA), the other half being generated by inversion symmetry. The dihedral angle between the adenine and phenyl ring planes is 26.71\u2005(7)\u00b0. The N6-benzoyl\u00adadenine mol\u00adecule crystallizes in the N(7)\u2014H tautomeric form with three non-protonated N atoms. This tautomeric form is stabilized by intra\u00admolecular N\u2014H\u22efO hydrogen bonding between the carbonyl (C=O) group and the N(7)\u2014H hydrogen atom on the Hoogsteen face of the purine ring, forming an S(7) ring motif. The two carboxyl groups of adipic acid inter\u00adact with the Watson\u2013Crick face of the BA mol\u00adecules through O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds, generating an R22(8) ring motif. The latter units are linked by N\u2014H\u22efN hydrogen bonds, forming layers parallel to (10-5). A weak C\u2014H\u22efO hydrogen bond is also present, linking adipic acid mol\u00adecules in neighbouring layers, enclosing R22(10) ring motifs and forming a three-dimensional structure. C=O\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions are also present in the structure.The asymmetric unit of the title co-crystal, C As evident from the angles at N7 [C8\u2014N7\u2014C5 = 106.82\u2005(11)\u00b0] and N9 [C8\u2014N9\u2014C4 = 103.90\u2005(11)\u00b0], the N6-benzoyl\u00adadenine moiety exists in the N(7)\u2014H tautomeric form with non-protonated N1, N3 and N9 atoms. In addition, the C8\u2014N7 bond [1.3415\u2005(17)\u2005\u00c5)] is longer than C8\u2014N9 [1.3175\u2005(19)\u2005\u00c5]. These values are similar to those in neutral N6-benzoyl\u00adadenine ring motif. The dihedral angle between the adenine and phenyl ring plane is 26.71\u2005(7)\u00b0 and the C6\u2014N6\u2014C10\u2014C11 torsion angle is 173.08\u2005(14)\u00b0. The bond lengths and bond angles of AA are in the range of values reported N6-benzyl\u00adadenine , N6-benz\u00adoyl\u00adadenine-dl-tartaric acid (1:1) is also present (Table\u00a01A functions as a bifurcated hydrogen-bond acceptor whereas N7\u2014H is a bifurcated hydrogen-bond donor.Each of the two carboxyl groups of adipic acid inter\u00adacts with the Watson\u2013Crick face (atoms N1 and N6) of the corres\u00adponding BA through O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds, generating an if Fig.\u00a01. Thus eas Table\u00a01 forming t Table\u00a01, linkinga) and C10=O1B\u22ef\u03c0 inter\u00adactions between the carbonyl oxygen O1B and the centroid of the (N1/C2/N3/C4/C5/C6) pyrimidine ring , N6-benzoyl\u00adadenine-dl-tartaric acid (1:1) and adipic acid (19\u2005mg) . The mixture was warmed in a water bath for 20 min. After cooling to room temperature, colourless plate-like crystals were collected from the mother liquor after a few days (m.p. 438\u2005K).The title co-crystal was synthesized by mixing a DMF solution of Uiso(H) = kUeq, where k = 1.5 for hy\u00addroxy and 1.2 for all other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016007581/hg5474sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016007581/hg5474Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016007581/hg5474Isup3.cmlSupporting information file. DOI: 1478504CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Zn\u2014Cl bond lengths differ sligthly at 2.2310\u2005(10) and 2.2396\u2005(11)\u2005\u00c5 while the Zn\u2014S bond lengths are equal at 2.3663\u2005(9) and 2.3701\u2005(10)\u2005\u00c5. The Cl\u2014Zn\u2014Cl angle is 116.04\u2005(4) and S\u2014Zn\u2014S is 101.98\u2005(3)\u00b0. All other angles at the central Zn atom range from 108.108\u2005(3) to 110.21\u2005(4)\u00b0. The C\u2014S\u2014Zn angles are 100.75\u2005(10) and 103.68\u2005(11)\u00b0, the difference most probably resulting from packing effects, as both the C\u2014S and both the S\u2014Zn bonds are equal in each case. The two imidazole ring planes make a dihedral angle of 67.9\u2005(1)\u00b0. The CH3 groups of one isopropyl moiety are disordered over two sets of sites with occupation factors of 0.567\u2005(15) and 0.433\u2005(15). It may be noteworthy that the isomolecular Cu complex shows a different crystal packing (group\u2013subgroup relation) with the Cu atom lying on a twofold rotation axis. In the crystal, the shortest non-bonding contact is a C\u2014H\u22efCl inter\u00adaction. This leads to the formation of centrosymmetric dimers that are stacked along the c-axis.The mol\u00adecular structure of the title compound, [ZnCl DOI: 10.1107/S1600536814023642/hp2069Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814023642/hp2069fig1.tif. DOI: Mol\u00adecular structure of the title compound with anisotropic displacement parameters drawn at the 50% probability level. Both orientations of disordered isopropyl group at C13 shown.1031227CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the mol\u00adecules are linked through weak C\u2014H\u22efCl hydrogen bonds into a tape structure.In the title compound, [Pd(C 2(C11H9N)(C13H14N2)], represents a new class of palladium-based polymerizable monomer which could give a potentially catalytically active polymer. It was synthesized via transmetallation from the corresponding silver complex. The PdII ion coordinates two Cl anions, one C atom from the N-heterocyclic carbene (NHC) ligand and one N atom from the 4-phenyl\u00adpyridine ligand, displaying a slightly distorted square-planar geometry. The dihedral angle between the imidazole ring and the pyridine ring is 34.53\u2005(8)\u00b0. The Pd\u2014C bond length between the NHC ligand and the PdII ion is 1.9532\u2005(16)\u2005\u00c5. In the crystal, weak non-classical C\u2014H\u22efCl hydrogen bonds link the mol\u00adecules into a tape structure along [101]. A weak \u03c0\u2013\u03c0 inter\u00adaction is also observed [centroid\u2013centroid distance = 3.9117\u2005(11)\u2005\u00c5]. The title compound, [PdCl The two mutually trans Cl ions fulfil the coordination sphere and 65.78\u2005(7)\u00b0, respectively. The C12\u2014C13 bond length of the vinyl group is 1.299\u2005(3)\u2005\u00c5, corroborating the double-bond character. The same goes for the C2\u2014C3 distance which is 1.330\u2005(3)\u2005\u00c5. The N2\u2014C4\u2014Pd1\u2014N3, N1\u2014C4\u2014Pd1\u2014Cl2, C18\u2014N3\u2014Pd1\u2014Cl2 and C17\u2014C16\u2014C19\u2014C24 torsion angles are \u221230\u2005(7), 81.15\u2005(15), \u221249.40\u2005(15) and 32.42\u2005(3)\u00b0, respectively. A Cambridge Structural Database (CSD) search to validate the Pd\u2014Cl and Pd\u2014N bonding was performed over 47 entries. The Cl\u2014Pd\u2014Cl and N\u2014C\u2014N angles range from 170 to 180\u00b0 and from 104.8 to 106.2\u00b0, respectively; the Pd\u2014Cl bond lengths are in the range 2.286\u20132.318\u2005\u00c5. The bond lengths and angles of the title compound 4 shows a \u03c0\u2013\u03c0 inter\u00adaction between the C19\u2013C24 phenyl rings of neighbouring mol\u00adecules with a centroid\u2013centroid distance of 3.9117\u2005(11)\u2005\u00c5 route, as shown in Fig.\u00a042 gave the chlorido-bridged palladium dimer 3. Cleavage of the dimer with phenyl\u00adpyridine afforded complex 4 in excellent yield. With its vinyl groups it can serve as a precursor in co-polymerization reactions with e.g. styrene to form polymeric materials with catalytic properties.2(bmim)]2[PdCl (4). 4-Phenyl\u00adpyridine was added to a 40\u2005ml solution of 3 in dry CH3CN and stirred at ambient temperature for 24\u2005h, during which time the solution changed colour to clear yellow. The mixture was filtered through Celite and all solvents were evaporated. The solids were dissolved in CH2Cl2 and, upon addition of n-hexane, a yellow solid was formed, which was collected on a frit and dried under vacuum to give 0.153\u2005g (93%) of 4.4 suitable for X-ray diffraction were obtained by slow diffusion of n-hexane into a saturated CH2Cl2 solution of the compound.Single crystals of Uiso(H) = 1.2Ueq(C).Crystal data and structure refinement details are summarized in Table\u00a0210.1107/S2056989016004394/is5447sup1.cifCrystal structure: contains datablock(s) Global, I. DOI: 10.1107/S2056989016004394/is5447Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016004394/is5447sup3.tifNumbering for the assignment of NMR spectra. DOI: 1468135CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II, four aqua and two N-bound 2-chloro\u00adpyrazine ligands, weak O\u2014H\u22efO as well as \u03c0\u2013\u03c0 inter\u00adactions play important roles in the mol\u00adecular self-assembly, resulting in the formation of a three-dimensional structure.Between the tosyl\u00adate anions and the octa\u00adhedral complex cations consisting of Fe II(C4H3ClN2)2(H2O)4](C7H7O3S)2, contains a complex cation with point group symmetry 2/m. The high-spin FeII cation is hexa\u00adcoordinated by four symmetry-related water and two N-bound 2-chloro\u00adpyrazine mol\u00adecules in a trans arrangement, forming a distorted FeN2O4 octa\u00adhedron. The three-dimensional supra\u00admolecular structure is supported by inter\u00admolecular O\u2014H\u22efO hydrogen bonds between the complex cations and tosyl\u00adate anions, and additional \u03c0\u2013\u03c0 inter\u00adactions between benzene and pyrazine rings. The methyl H atoms of the tosyl\u00adate anion are equally disordered over two positions.The title salt, [Fe Series of thio\u00adcyanato coordination polymers [M(NCS)2L2]\u221e in which the small-sized thiocyanate anions are terminally N-bound and therefore not involved in any magnetic exchange interactions are also known 2(H2O)4](C7H7O3S)2 containing a cationic iron(II) complex with 2-chloro\u00adpyrazine and aqua ligands, and tosyl\u00adate as an anion.In this paper, we report on the crystal structure of [Fe2(H2O)4]2+ and two tosyl\u00adate anions. The FeII atom, located on a special position with site symmetry 2/m, is sixfold coordinated by two N atoms of two symmetry-related 2-chloro\u00adpyrazine ligands occupying the axial positions and four O atoms of four H2O mol\u00adecules forming the equatorial plane \u2005\u00c5] of the H2O mol\u00adecules are significantly shorter than those between FeII and N [2.200\u2005(2)\u2005\u00c5] atoms of the two 2-chloro\u00adpyrazine ligands, hence the resulting FeO4N2 octa\u00adhedron is distorted. The metal-to-ligand distances clearly signalize the high-spin nature of the complex described in here \u00b0; for symmetry codes see caption to Fig.\u00a01ii\u2014Fe1\u2014N1 = 90.68\u2005(5)\u00b0] indicate only a small angular distortion.The structure of the title compound consists of a complex cation [Fe(2-chloro\u00adpyrazine)ne Fig.\u00a01. The disA\u22efO2 and O1\u2014H1B\u22efO3i hydrogen bonds (OTs = p-toluene\u00adsulfonate) and ascorbic acid (0.001\u2005g) in water (5\u2005ml). After seven days this yielded colourless blocks of the title compound that were collected, washed with water and dried in air. Yield 0.090\u2005g (64%).Crystals of the title compound were obtained by adding 2-chloro\u00adpyrazine to Fe(OTs)d(C\u2014H) = 0.95\u2005\u00c5 for aromatic and 0.98\u2005\u00c5 for CH3 hydrogen atoms. Because of the symmetry of the complete complex cation, methyl H atoms were modelled as equally disordered over two sets of sites. The H atoms of the water mol\u00adecule were located from a difference Fourier map and were modelled with a common isotropic displacement parameter fixed at 0.08\u2005\u00c52. The O\u2014H bonds lengths were constrained to 0.82\u2005\u00c5. The Uiso values were constrained to be 1.5Ueq of the carrier atom for methyl H atoms and 1.2Ueq for the remaining H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015010713/wm5168sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015010713/wm5168Isup2.hklStructure factors: contains datablock(s) I. DOI: 1404715CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure consists of discrete tetra\u00adhedral complexes, that are linked by inter\u00admolecular N\u2014H\u22efO, C\u2014H\u22efO and N\u2014H\u22efO hydrogen bonding. 2(C6H6N2O)2], consists of one Zn2+ cation located on a twofold rotation axis, as well as of one thio\u00adcyanate anion and one neutral isonicotinamide ligand, both occupying general positions. The Zn2+ cation is tetra\u00adhedrally coordinated into a discrete complex by the N atoms of two symmetry-related thio\u00adcyanate anions and by the pyridine N atoms of two isonicotinamide ligands. The complexes are linked by inter\u00admolecular C\u2014H\u22efO and N\u2014H\u22efO, and weak inter\u00admolecular N\u2014H\u22efS hydrogen-bonding inter\u00adactions into a three-dimensional network.The asymmetric unit of the title complex, [Zn(SCN) The synthesis and crystal structure of the resulting compound, [Zn(NCS)2(C6H6N2O)2], are reported here.The synthesis of magnetic materials is still a major field in coordination chemistry to 123.6\u2005(2)\u00b0.The asymmetric unit of the title compound consists of one Znex Fig.\u00a01. As expeex Fig.\u00a01. The angc axis and are linked by inter\u00admolecular N\u2014H\u22efO hydrogen bonding between one of the two amide H atoms and the amide O atom of a neighboring complex 2 was synthesized by stirring 3.076\u2005g Ba(NCS)2\u00b73H2O (10\u2005mmol) with 1.795\u2005g ZnSO4\u00b7H2O (10\u2005mmol) in 350\u2005ml water. The white residue was filtered off and the filtrate was dried using a rotary evaporator. The homogenity was checked by X-ray powder diffraction and elemental analysis. Crystals of the title compound suitable for single crystal X-Ray diffraction were obtained by the reaction of 27.2\u2005mg Zn(NCS)2 (0.15\u2005mmol) with 36.64\u2005mg isonicotinamide (0.3\u2005mmol) in methyl\u00adcyanide (1.5\u2005ml) within a few days.Ba(NCS)Uiso(H) = 1.2Ueq using a riding model with C\u2014H = 0.95\u2005\u00c5 for aromatic and N\u2014H = 0.88\u2005\u00c5 for the amide H atoms. The absolute structure was determined and is in agreement with the selected setting [Flack x parameter: 0.005\u2005(19) by classical fit to all intensities I. DOI: 10.1107/S2056989016008963/wm5297Isup2.hklStructure factors: contains datablock(s) I. DOI: 1483379CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III atoms, the title compound, [Re2Cl2{O2C(CH2)2CH3}4], exhibits a paddle-wheel or lantern-type structure with four n-butyrate groups bridging two ReIII atoms in a syn\u2013syn fashion. The axial chloride ligands together with the Re\u2014Re quadruple bond [2.2330\u2005(3)\u2005\u00c5] complete an essentially octa\u00adhedral geometry around each ReIII atom. There is little distortion, with an Re\u2014Re\u2014Cl bond angle of 176.18\u2005(3)\u00b0 and typical cis-O\u2014Re\u2014O bond angles ranging from 89.39\u2005(11) to 90.68\u2005(11)\u00b0. There are two mol\u00adecules in the unit cell, and no significant inter\u00admolecular inter\u00adactions were noticed between mol\u00adecules in the crystal.With an inversion center at the mid-point of the two Re DOI: 10.1107/S1600536814020273/wm5055Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814020273/wm5055fig1.tifx y z . DOI: x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a02.]The mol\u00adecular structure of the title compound with displacement ellipsoids drawn at the 50% probability level. Hydrogen atoms are drawn as small spheres of arbitrary radius. [Symmetry code i) \u2212Click here for additional data file.10.1107/S1600536814020273/wm5055fig2.tif. DOI: The packing diagram for the title compound viewed along [100].1023523CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Both complexes, crystallized from aceto\u00adnitrile and diethyl ether, exhibit paddlewheel conformations with quadruple bonds between the Re atoms.The structures of [Re 2Br2(O2CC3H7)4], (1), and [Re2(O2CC3H7)4Cl2]\u00b72CH3CN, (2), both exhibit paddlewheel structures with four carboxyl\u00adate ligands bridging two ReIII atoms. The Re\u2014Re distances are 2.2325\u2005(2) and 2.2299\u2005(3)\u2005\u00c5, indicating quadruple bonds between the ReIII atoms in each complex. Both complexes contain an inversion center at the mid-point of the Re\u2014Re bond. The Re\u2014Br bond [2.6712\u2005(3)\u2005\u00c5] in (1) is 0.1656\u2005(6)\u2005\u00c5 longer than the Re\u2014Cl distance [2.5056\u2005(5)\u2005\u00c5] of (2). In (2), the N atom of each co-crystallized aceto\u00adnitrile solvent mol\u00adecule is nearly equidistant between and in close contact with two carboxyl\u00adate C atoms.The title complexes, [Re The first compound discovered to contain a metal\u2013metal quadruple bond was K al. 1966 and in s al. 2001, Cotton al. 1997, and Veg al. 2002. This co1) and (2), respectively, are indicative of quadruple bonds and 2.2299\u2005(3)\u2005\u00c5, in ( Tables 1 and 2 \u25b8.2) also contains one co-crystallized aceto\u00adnitrile solvent mol\u00adecule in a general position, thus giving rise to twice that in the formula unit. The asymmetric unit of (X\u2014Re\u2014Re\u2014X bonds in (1) and (2) are nearly linear, as can be seen in the Re\u2014Re\u2014Br [175.018\u2005(7)\u00b0] and Re\u2014Re\u2014Cl [178.254\u2005(11)\u00b0] bond angles, and are comparable to those observed in similar compounds is similar to those of the previously published analog without co-crystallized aceto\u00adnitrile 3)4], and [Re2Cl2(O2CC6H5)4]\u00b72CHCl3 is slightly longer than the Re\u2014Br bond [2.603\u2005(1)\u2005\u00c5] found in [Re2Br2(O2CC(CH3)3)4] and (2) differ by 0.1656\u2005(6)\u2005\u00c5 and those of Cotton and coworkers differ by 0.126\u2005(3), both of which are consistent with the difference in covalent radii of Cl and Br (0.15\u2005\u00c5).The 1) is isotypic with the chlorido analog published by Thomson et al. (20142Cl2(O2CC3H7)4]. In compound (2), the C1\u2014C2\u2014C3\u2014C4 torsion angle is \u221270.2\u2005(2)\u00b0, comparable to \u221267.9\u2005(2)\u00b0 for C5\u2014C6\u2014C7\u2014C8 4] without co-crystallizing solvent, the torsion angles vary more Table\u00a01.2) nitro\u00adgen atom N1 of the co-crystallized aceto\u00adnitrile solvent mol\u00adecule is located at distances of 3.197\u2005(3) and 3.216\u2005(3)\u2005\u00c5 from the carboxyl\u00adate carbon atoms C1 and C5, respectively. This is just within the sum of the van der Waals radii of 3.25\u2005\u00c5 = 1.2Ueq(C) and methyl, C\u2014H = 0.98\u2005\u00c5, with Uiso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015020563/pj2025sup1.cifCrystal structure: contains datablock(s) 1, 2, global. DOI: 10.1107/S2056989015020563/pj20251sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989015020563/pj20252sup3.hklStructure factors: contains datablock(s) 2. DOI: 1434257, 1434256CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the acetate anion is connected to the complex mol\u00adecule via a pair of N\u2014H\u22efO hydrogen bonds [graph-set motif R22(8)] and the solvent methanol mol\u00adecule is connected to the anion via an O\u2014H\u22efO hydrogen bond. This aggregate is further connected through a weak C\u2014H\u22efO hydrogen bond, forming a chain along [100]. In addition, sixfold phenyl embraces with inter\u00admolecular distances of 6.6463\u2005(13)\u20136.667\u2005(2)\u2005\u00c5 are arranged in a chain along [001]. The combination of hydrogen bonding and phen\u00adyl\u22efphenyl inter\u00adactions leads to the formation of a two-dimensional network parallel to (010).In the mononuclear title complex, [Ag(C For pot al. 2007; Isab et al. 2010. For det al. 2010; Scudder al. 2010. For hyd al. 1990. 5H12N2S)(C18H15P)3](C2H3O2)\u00b7CH4O = 0.053wR(F2) = 0.120S = 1.0016670 reflections653 parametersH-atom parameters constrainedmax = 1.23 e \u00c5\u22123\u0394\u03c1min = \u22121.13 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I. DOI: 10.1107/S1600536814010824/lh5701Isup2.hklStructure factors: contains datablock(s) I. DOI: 1002206CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecules associate through an inter\u00admolecular O\u2014H\u22efNnitrile hydrogen bond and are inter\u00adlinked through very weak C\u2014H\u22efN hydrogen bonds.In the crystal of the title substituted hemibi\u00adquinone derivative, the ring systems inter\u00adact through an intra\u00admolecular O\u2014H\u22efO 16H11BrN2O4 or [BrHBQH2(CN)2], the substituted benzene rings are rotated about the central C\u2014C bond, forming a dihedral angle of 53.59\u2005(7)\u00b0. The ring systems inter\u00adact through an intra\u00admolecular O\u2014H\u22efOmeth\u00adoxy hydrogen bond, which induces a geometry quite different from those in previously reported hemibi\u00adquinone structures. In the crystal, the mol\u00adecules associate through an inter\u00admolecular O\u2014H\u22efNnitrile hydrogen bond, forming chains which extend along [100] and are inter\u00adlinked through very weak C\u2014H\u22efN hydrogen bonds, giving a overall two-dimensional structure lying parallel to (010).In the crystal of the title substituted hemibi\u00adquinone derivative, C This bending of the mol\u00adecule about its long axis may also be due to hydrogen bonding as it causes the meth\u00adoxy group to approach the OH group more closely. The aromatic C\u2014C bonds of both rings have a narrow range of distances [from 1.387\u2005(2) to 1.412\u2005(2)\u2005\u00c5]. The C\u2014C, C\u2014O, C\u2014N, and C\u00a0N distances for the mol\u00adecule are similar to the corresponding distances in 2,3,5,6-tetra\u00adcyano\u00adhydro\u00adquinone \u00b0. As a result of the influence of other short contacts and supra\u00admolecular inter\u00adactions (see below), the phenolic C\u2014O\u2014H bond angles deviate when compared to the meth\u00adoxy C\u2014O\u2014C bond angles: C8\u2014O3\u2014H is 108\u2005(2)\u00b0, C11\u2014O4\u2014H is 112.3\u2005(2)\u00b0, C3\u2014O2\u2014C14 is 117.9\u2005(1)\u00b0, and C6\u2014O1\u2014C13 is 117.2\u2005(1)\u00b0. As in other structures, the meth\u00adoxy groups are aligned mostly in-plane with the benzene ring, C5\u2014 C6\u2014O1\u2014C13 being bent out of plane by \u22124.5\u2005(2)\u00b0 and C2\u2014C3\u2014O2\u2014C14 bent out of plane by \u22121.3\u2005(2)\u00b0. The C12\u2014C11\u2014O4\u2014H phenol group is also nearly planar, being bent out of plane by 1.3\u00b0. However, the hydrogen-bonded phenol is unsurprisingly bent out of plane, C7\u2014C8\u2014O3\u2014H = 38\u2005(2)\u00b0. The meth\u00adoxy methyl groups point away from the sterically restricting groups a-axis direction. Both nitrile groups are involved in inter\u00admolecular hydrogen-bonding inter\u00adactions, the first one (O4\u2014H\u22efN1) strong, the second one (C2\u2014H\u22efN2) weaker but still highly directional. For details, see Table\u00a01a- and b-axis directions, respectively, forming sheets parallel to (010) contacts to six neighboring mol\u00adecules Fig.\u00a03. As in p0) Fig.\u00a04. The quib and stacking along c can be seen in Fig.\u00a05ac plane.The remaining two mol\u00adecules in the unit cell are oriented orthogonally to the central mol\u00adecules. These mol\u00adecules are anti\u00adparallel to each other, where the di\u00admeth\u00adoxy\u00adbenzene rings stack with those of the central pair. Slightly repulsive \u03c0-inter\u00adactions between mol\u00adecules along 2O. Upon pouring the aqueous solution into the organic solution, the mixture immediately changed from a vibrant red to a deep purple. After stirring for 1\u2005h, 50\u2005\u00b5L of concentrated HCl solution was added, changing the color of the mixture from purple to bright orange. The mixture was diluted with 50\u2005mL of water and the aceto\u00adnitrile was removed by rotary evaporation. A tan\u2005powder precipitated, which was recovered by filtration and washed with water to yield the crude product. This material was recrystallized from acetone giving 0.196\u2005g (70.4%) of pure material as yellow\u2013orange prisms. 1H NMR \u03b4 = 10.02 , 8.75 , 7.34 , 7.24 , 7.05 , 3.88 , 3.82 .2-Bromo-5-cyclo\u00adhexa-2,5-diene-1,4-dione, BrHBQBr, was dissolved in 350\u2005mL of aceto\u00adnitrile. In a separate beaker, potassium cyanide was dissolved in 50\u2005mL of HUiso = 1.5Ueq(O). Hydrogen atoms bonded to carbon were placed in calculated positions with C\u2014H = 0.93\u2005\u00c5 (aromatic) or 0.96\u2005\u00c5 (meth\u00adyl) and their coordin\u00adates and thermal parameters were constrained to ride on the carrier atom, with Uiso = 1.5Ueq(aromatic C) or 1.5Ueq(methyl C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016005715/zs2361sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016005715/zs2361Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016005715/zs2361Isup3.cmlSupporting information file. DOI: 1472611CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-(4-methyl\u00adphen\u00adyl)benzene\u00adsulfonamide and N-(4-fluoro\u00adphen\u00adyl)-4-meth\u00adoxy\u00adbenzene\u00adsulfonamide, the supra\u00admolecular architecture of the former is controlled by C\u2014H\u22ef\u03c0ar\u00adyl inter\u00adactions, forming a two-dimensional architecture, while in the latter, a pair of C\u2014H\u22efO inter\u00admolecular inter\u00adactions lead to the formation of a three-dimensional architecture.In the crystal structures of 4-meth\u00adoxy- N-(ar\u00adyl)aryl\u00adsulfonamides, namely, 4-meth\u00adoxy-N-(4-methyl\u00adphen\u00adyl)benzene\u00adsulfonamide, C14H15NO3S, (I), and N-(4-fluoro\u00adphen\u00adyl)-4-meth\u00adoxy\u00adbenzene\u00adsulfonamide, C13H12FNO3S, (II), were determined and analyzed. In (I), the benzene\u00adsulfonamide ring is disordered over two orientations, in a 0.516\u2005(7):0.484\u2005(7) ratio, which are inclined to each other at 28.0\u2005(1)\u00b0. In (I), the major component of the sulfonyl benzene ring and the aniline ring form a dihedral angle of 63.36\u2005(19)\u00b0, while in (II), the planes of the two benzene rings form a dihedral angle of 44.26\u2005(13)\u00b0. In the crystal structure of (I), N\u2014H\u22efO hydrogen bonds form infinite C(4) chains extended in [010], and inter\u00admolecular C\u2014H\u22ef\u03c0ar\u00adyl inter\u00adactions link these chains into layers parallel to the ab plane. The crystal structure of (II) features N\u2014H\u22efO hydrogen bonds forming infinite one dimensional C(4) chains along [001]. Further, a pair of C\u2014H\u22efO inter\u00admolecular inter\u00adactions consolidate the crystal packing of (II) into a three-dimensional supra\u00admolecular architecture.Crystal structures of two The dihedral angle between the two parts of disordered benzene ring, i.e. C1/C2A/C3A/C4/C5A/C6A and C1/C2B/C3B/C4/C5B/C6B, is 28.0\u2005(1)\u00b0. The dihedral angle between the sulfonyl benzene ring (considering the major component) and the aniline ring is 63.36\u2005(19)\u00b0, and the N\u2014C bond in the C\u2014SO2\u2014NH\u2014C segment has a gauche torsion with respect to the S=O bonds. Further, the mol\u00adecule is twisted at the S\u2014N bond, with a C1\u2014S1\u2014N1\u2014C7 torsion angle of 66.33\u2005(19)\u00b0. The meth\u00adoxy group in the sulfonyl\u00adbenzene ring is in the same plane as that of the major component of the disordered sulfonyl\u00adbenzene ring, the torsion angle C5A\u2014C4\u2014O3\u2014C14 being \u2212176.2\u2005(4)\u00b0, while it deviates slightly from planarity with respect to the minor component, the C5B\u2014C4\u2014O3\u2014C14 torsion angle being 165.9\u2005(4)\u00b0.In (I)2\u2014NH\u2014C segment has a gauche torsion with respect to the S=O bonds. Further, the mol\u00adecule is twisted at the S\u2014N bond, with a C1\u2014S1\u2014N1\u2014C7 torsion angle of 68.4\u2005(2)\u00b0. Similar to (I)In (II)C(4) chains along [010]. Neighbouring C(4) chains are inter\u00adconnected via C\u2014H\u22ef\u03c0ar\u00adyl inter\u00adactions s Table\u00a01 into lays Table\u00a01 parallelC(4) chains along [001]. Further, weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions s Table\u00a02 consolids Table\u00a02.N-(4-substituted-phen\u00adyl)-4-meth\u00adoxy\u00adbenzene\u00adsul\u00adfon\u00adam\u00adides benzene\u00adsulfonamide, (III), 4-meth\u00adoxy-N-(4-meth\u00adoxy\u00adphen\u00adyl)benzene\u00adsulfonamide, (IV), and N-(4-chloro\u00adphen\u00adyl)-4-meth\u00adoxy\u00adbenzene\u00adsulfonamide, (V), have been reported previously. Compounds (IV) and (V) crystallize in monoclinic syngony, while compound (III) crystallizes in ortho\u00adrhom\u00adbic syngony. The dihedral angles between the two benzene rings in (III), (IV) and (V) are 55.1\u2005(1), 56.3\u2005(1) and 42.6\u2005(1)\u00b0, respectively. Comparison of the dihedral angles between the two benzene rings in (I)\u2013(V) shows that, when an electron-donating substituent is introduced into the para position of the aniline ring of (I)Three para position of the aniline ring of (I)C(4) chains, and thus, the supra\u00admolecular architecture is one-dimensional. In (IV), one N\u2014H\u22efO hydrogen bond and two alternating C\u2014H\u22ef\u03c0ar\u00adyl (centroid of aniline ring) inter\u00adactions direct a two-dimensional architecture. This is quite similar to the crystal structure of (I)C(4) chains. Further, (V) does not feature any structuredirecting inter\u00admolecular inter\u00adactions, and thus, the structure is one-dimensional. In contrast to this, the crystal structure of (II)Comparison of the crystal structures (I)et al. (2015Compounds (I) al. 2015. The purR1, wR2, and S (goodness-of-fit), a partially obscured reflection (i.e. 100) was omitted from the final refinement of (I)A and B) of the disordered benzene\u00adsulfonyl ring in (I)A) and 0.054\u2005(1)\u2005\u00c5 (minor-component ring B). The disordered atoms in both components were isotropically refined, and the C\u2014C bond lengths were restrained to 1.391\u2005(1)\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015019787/cv5497sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989015019787/cv5497Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015019787/cv5497Isup4.cmlSupporting information file. DOI: 10.1107/S2056989015019787/cv5497IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989015019787/cv5497IIsup5.cmlSupporting information file. DOI: 1432501CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The SrII ion, situated on a C2 axis, is coordinated by four O atoms from four pydc2\u2212 ligands and four water mol\u00adecules. The coordination geometry of the SrII atom can be best described as a distorted dodeca\u00adhedron. Each SrII ion bridges four [Co(pydc)2(H2O)2]2\u2212 units by four COO\u2212 groups of four pydc2\u2212 ligands to form a three-dimensional network structure. Two additional solvent water mol\u00adecules are observed in the crystal structure and are connected to the three-dimensional coordination polymer by O\u2014H\u22efO hydrogen bonds. Further intra- and intermolecular O\u2014H\u22efO hydrogen bonds consolidate the overall structure.In the title polymeric complex, {[CoSr(C DOI: 10.1107/S2056989015014942/im2469Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015014942/im2469fig1.tifx y z x y z x y z x y z x y z . DOI: x,-y\u00a0+\u00a01,-z\u00a0+\u00a01; B: \u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a01; C: x\u00a0\u2212\u00a0y\u00a0+\u00a01/2,z; G: \u2212x,y,-z\u00a0+\u00a0x\u00a0\u2212\u00a0y,z\u00a0\u2212\u00a01/2.The mol\u00adecular structure of the title compound. Displacement ellipsoids are drawn at the 50% probability level. Symmetry codes: A: \u2212Click here for additional data file.10.1107/S2056989015014942/im2469fig2.tifb . DOI: b-axis, with hydrogen bonds drawn as dashed lines.The packing diagram for the title compound, viewed down the 761895CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Herein, we report X-ray structures of organic crystals that feature a chloride anion bound exclusively by hydrophobic Cali\u2013H groups. An X-ray structure of imidazolium-based scaffolds using Cali\u2013H\u00b7\u00b7\u00b7A\u2212 interactions (A\u2212\u2009=\u2009anion) shows that a halide anion is directly interacting with fifteen Cali\u2013H groups . Additional supporting interactions and/or other binding sites are not observed. We note that such types of complexes may not be rare since such high numbers of binding sites for an anion are also found in analogous tetraalkylammonium complexes. The Cali\u2013H\u00b7\u00b7\u00b7A\u2212 interactions are driven by the formation of a near-spherical dipole layer shell structure around the anion. The alternating layers of electrostatic charge around the anion arise because the repulsions between weakly positively charged H atoms are reduced by the presence of the weakly negatively charged C atoms connected to H atoms.Since the aliphatic C\u2013H\u00b7\u00b7\u00b7anion interaction is relatively weak, anion binding using hydrophobic aliphatic C\u2013H (C Because the Cali\u2013H\u00b7\u00b7\u00b7A\u2212 interaction is weak, anion binding by the Cali\u2013H groups is generally enhanced through the incorporation of additional binding sites to the anion. Indeed, though C\u2013H\u00b7\u00b7\u00b7A\u2212 type interactionsali\u2013H groups are rarely used as H-bond donors in synthetic receptors. Nevertheless, such interactions are essential to the overall stability of complexes of proteins and DNA as well as various organic transformations and the transition states of diverse catalytic cycles2ali\u2013H donors constitutes a useful opportunity to tailor molecular recognition phenomena4ali\u2013H\u00b7\u00b7\u00b7A\u2212 interactions due to relatively low binding strengths, although there are some reports about interactions between anions and multi Cali\u2013H donors67910\u2212 interactions. In comparison, the C\u2013H bonds found in nonpolar alkanes are relatively less acidic, though the corresponding interactions with anions may be increased through the incorporation of electron-withdrawing groups.The structure of an anion that is surrounded exclusively by multiple Cali\u2013H\u00b7\u00b7\u00b7A\u2212 H-bondsali\u2013H\u00b7\u00b7\u00b7A\u2212 hydrogen bonding was reported for adducts of anions and a resorcinarene cavitand, which was shown to adopt a bowl-shaped cavity that provided up to four convergent C\u2013H groups activated by adjacent oxygen atomsali\u2013H hydrogen bonding appear to be limited to a palladium complexali\u2013H groups1516\u2212 interaction sites towards an anion.The first charge-neutral systems of highly fluorinated receptors with aliphatic methylene groups were polarized by neighboring electronegative O and F atomsali\u2013H groups. The methyl and methylene moieties in this host form Cali\u2013H\u00b7\u00b7\u00b7A\u2212 interactions with various anions. In particular, we report that bisimidazolium and tetraalkylammonium based hosts drive the formation of multideca to pentadeca binding sites with halide anions exclusively through Cali\u2013H\u00b7\u00b7\u00b7A\u2212 interactions.Herein, we disclose the synthesis and study of a bis-imidazolium host bearing acidic C1](PF6)2 was synthesized and characterized using NMR spectroscopy were grown under anhydrous conditions. Single crystal X-ray diffraction analysis \u2009=\u20091.2U. The Cl\u2212 anion appeared to be surrounded by multiple Cali\u2013H groups di-hexafluorophosphate (Cl)2 are not small .Then, in our crystal structures, we find that these H\u2009\u2265\u2009100\u00b0) . It shou\u2212 in bis-tetramethylimidazolium complexes, we were pleasantly surprised to find that one of two kinds of Cl\u2212 has 15 binding sites via H-bonds and H-bonding-like interactions, ten strong H-donors ), 1 moderate H-donor (#13), and four weak H-donors (\u2212\u00b7\u00b7\u00b7C distances (dCl\u00b7\u00b7\u00b7C), twelve C atoms were within H-bond distance from Cl\u2212 (4.01\u20134.08\u2009\u00c5), in consideration that rvdWCl\u00b7\u00b7\u00b7(H)C: 4.01\u20134.08\u2009\u00c5 , while additional two C atoms were positioned at relatively long distances . The Cali\u2013H\u00b7\u00b7\u00b7Cl\u2212 angles (\u03b8) were measured to be in all circumstances greater than 148\u00b0 except for the bidentate case (129/124\u00b0 and 100/100\u00b0 in X-ray/DFT) by the two H atoms attached on the same C atom at dCl\u00b7\u00b7\u00b7C\u2009=\u20094.05\u2009\u00c5. Namely, eleven C atoms involved in strong bidentate H-bonds were in agreement with the formation of bona fide H-bonds. In addition, one C atom involved in bidentate Cali\u2013H\u00b7\u00b7\u00b7Cl\u2212 interactions with its two H atoms was considered to have weak H-bonding-like interactions (\u03b8: 129\u00b0 and 100\u00b0) because the dCl\u00b7\u00b7\u00b7H distances are somewhat long even though dCl\u00b7\u00b7\u00b7C (=\u20094.05\u2009\u00c5) \u2248 rvdWCl\u00b7\u00b7\u00b7(H) (=\u20094.01\u20134.08\u2009\u00c5). Nevertheless, these bidentate H-bonding-like interactions could be considered as bidentate H-bonds or at least bidentate H-bonding-like bindings. Further, two weak uni-dentate H bonding-like interactions at relatively long distances (dCl\u00b7\u00b7\u00b7C\u2009=\u20094.71/4.83\u2009\u00c5) but with favorable angles (\u03b8: 153\u00b0/177\u00b0) can also be taken into account as direct binding sites. Then, Cl shows fifteen binding sites (thirteen uni-dentate and one bidentate H-bonding or H-bonding-like interaction sites) by the Cali\u2013H groups in seven molecules of [1] surrounding the Cl\u2212 anion. Furthermore, each Cali\u2013H\u00b7\u00b7\u00b7Cl\u2212 binding for the given dCl\u00b7\u00b7\u00b7H and \u03b8 shows adequate BEs for all the fifteen cases due to the strong electrostatic interactions between negatively charged Cl\u2212 and positively charged H3C-NH3+. In all the above cases of fifteen H atoms, no other atom exists within the sphere having the diameter from each H atom to Cl\u2212, and thus, the fifteen H atoms directly bind Cl\u2212 as Voronoi nearest neighbors.From the analysis of binding sites for Cl 14, 15) . In term\u2212\u00b7\u00b7\u00b7H-Cali is mainly an electrostatic interaction between positive and negative charges where the positive charge of H is enhanced due to the polarization through the H-Cali bond. In the model system of Cl\u2212\u00b7\u00b7\u00b7H-CH3, the induction and electrostatic energies based on DFT are dominant for the binding in the fully optimized structure . Even with a highly increased distance between H and Cl, the induction energy gives significant contribution for binding: \u2206Etotal\u2009=\u2009\u22122.33; \u2206Eelst\u2009=\u2009\u22120.36; \u2206Eind\u2009=\u2009\u22120.95; \u2206Eexch\u2009=\u2009\u22120.36\u2009kcal/mol for dCl\u00b7\u00b7\u00b7H\u2009=\u20093.871\u2009\u00c5 than in the methane model (+0.218). All of the Cl\u2212\u00b7\u00b7\u00b7H-Cali angles deviate slightly from 180\u00b0 but, by considering flexibility of the range of H-bond angles, the angles in the 15 H atoms cases are in favor of significant interaction energies. In addition, if the angle reduces from 180\u00b0 to 100\u00b0, the Hx-C (Hx: hydrogen interacting with Cl\u2212) bond distance decreases from 1.108 to 1.102\u2009\u00c5, while Cl\u2212\u00b7\u00b7\u00b7Hx distance increases from 2.705 to 3.5\u2009\u00c5. Namely, as the angle is close to 180\u00b0, the Cl\u2212\u00b7\u00b7\u00b7Hx interaction energy becomes stronger, showing the significant angle-dependence of the H-bonding characteristics.Although we classified the above binding interaction as the H-bonding and H-bonding-like interactions, even the latter show most features of H-bond addressed in the IUPAC provisional recommendation (criteria and characteristics)20\u2212\u00b7\u00b7\u00b7H\u2013Cali interactions in the CH4\u2013Cl\u2212 and NH3CH3+\u00b7\u00b7\u00b7Cl\u2212 models as well as the crystal structure of [1](Cl)2 have H-bond-like characteristics based on both the IUPAC recommendation and the Koch-Popelier definition (\u03c1(r)\u2009=\u20090.002\u20130.040 au and 1](Cl)2 crystal structure highlights that Cl\u2212 is stabilized by strong H-bonds as well as strong ionic electrostatic interactions (dCl\u00b7\u00b7\u00b7Hx\u2009\u2264\u20093.15\u2009\u00c5 in X-ray structure) and weak H-bonding-like interactions critical pointractions . Overallali-H groups is conceptually similar to the cation recognition of hydrophobic aromatic rings, but the origin of noncovalent interactions are quite different for such high number of binding sites similar to those described herein. Therein, we indeed found 14 binding sites for anions in the tetraalkylammouniun complexes (BUXTOD)actions) , and SI,3 groups that were bonded to the N atoms, where the C atom was slightly positively charged due to the ammonium group. Thus, weakly positively charged H atoms surround the anion nearly spherically, and then the weakly negatively charged C atoms surround the region of positively charged H atoms. The nearly concentric structures showing alternating +/\u2212 electrostatic potential shells are formed 13dCl\u00b7\u00b7\u00b7H\u2009=\u20092.88\u20133.13\u2009\u00c5, dCl\u00b7\u00b7\u00b7C\u2009=\u20093.65\u20133.75\u2009\u00c5, \u03b8Cl\u00b7\u00b7\u00b7H\u2013C\u2009=\u2009121\u2013132\u00b0; dI\u00b7\u00b7\u00b7H\u2009=\u20093.35\u20133.30\u2009\u00c5, dI\u00b7\u00b7\u00b7C\u2009=\u20093.88\u2009\u00c5, \u03b8I\u00b7\u00b7\u00b7H\u2013C\u2009=\u2009117\u2013120\u00b0]. In consideration of small angles, this dodeca coordination somewhat reflects a caged structure, i.e., strained coordination, instead of H-bonding. However, it may still be labeled as a dodeca-coordination complex, with centrospherical shells that feature Cali\u2013H\u00b7\u00b7\u00b7A\u2212 interactions and contain alternating regions of +/\u2212 electrostatic potential.Such nearly centrospherical shell structures containing multiple C\u2013H\u00b7\u00b7\u00b7A\u2212 anions in the bisimidazolium complex have pentadeca binding sites exclusively by Cali\u2013H groups. The electrostatic potential maps feature nearly concentro-spherical shells with alternating +/\u2212 electrostatic interactions, quite different in structure from many well-known H-bond complexes for anions. These intriguing Cali\u2013H\u00b7\u00b7\u00b7A\u2212 interactions have not been properly recognized previously. The positively charged hosts render the aliphatic C\u2013H moieties relatively acidic and thus increase their binding affinities for anions. Collectively, the results described herein may give rise to new classes of aliphatic hosts that display selectivity toward anions via tight control of cavity geometry and acidity of their respective H-bond donors. We also note that complexes of tetradeca binding sites and other multiple binding sites show nearly concentro-spherical shells depicting alternating +/\u2212 electrostatic potential for the Cali\u2013H\u00b7\u00b7\u00b7A\u2212 interactions.Our study finds that the ClHere, we discuss the number of binding sites as compared with the coordination number (CN) which are well defined in inorganic chemistry. In solid and liquid states materials, the CN is also often used, differently from the terminology used in inorganic chemistry. Indeed, the CN has been a widely used terminology in various branches of science. The definition of CN originates from mathematics, where it means the number of equivalent hyperspheres in n dimensions that can touch an equivalent hypersphere without any intersections45Though the definition might not be clear in certain cases, it is assumed that each binding event needs to show significant positive BE associated with noncovalent interaction, noncovalent bonding, or electrostatic interaction. The van der Waals (vdW) interactions are generally not considered in counting CNs. Coordination phenomena often include cation-anion interactions, hydrogen bonding, and \u03c0 interactions4748\u03b8 between two vectors constructed from any other atom to the coordinating site and the coordinated site. If any other atom has the angle \u03b8\u2009>\u200990\u00b0, the assumed coordinating atom/site should be removed from the direct coordination. If all angles \u03b8 obtained for all other atoms are less than 90\u00b0, it means that all of them are out of the sphere, and so this case is considered to be directly coordinated. In this way, the obtained CN is equivalent to the Voronoi-based CN nearest neighborsWhen the local coordination does not have high symmetry, the direct coordination can be considered as a case where a given coordinating atom (or site) does not have any other atoms in the sphere centered on the midpoint between the given site and the coordinated atom. This can be analyzed by measuring the angle 4)4]152+, which has a large metal cation and many small ligands3BNMe2BH3)4]4)4, its derivatives 52544 groups.Although examples of complexes that feature multiple coordination numbers have been widely observed, cases of very high coordination are rare. Structures with very high coordination numbers have been found with actinide-based cations, such as + 905.500, found 905.070; m/z calcd. for [M \u2013 I]+ 389.298, found 388.914.1,2,4,5-Tetramethylimidazole and diiodomethane were heated at 110\u2009\u00b0C in a sealed tube overnight. The resulting mixture was filtered and washed with dichloromethane several times to afford [1](I)2 was added a saturated aqueous solution of NH4PF6 at room temperature. A precipitate formed which was subsequently collected by filtration and washed with water to afford [1](PF6)2 as a light brown solid. Yield: 2.16\u2009g, 83%. 1H NMR , \u03b4 6.15 , 3.61 , 2.53 , 2.20 , 2.04 . 13CNMR , \u03b4 145.83 (N\u2009=\u2009CCH3N), 129.44 (H3CN-CCH3\u2009=\u2009C), 126.87 (-CH2N-CCH3\u2009=\u2009C), 56.41 (CH2), 33.77 (NCH3), 11.88 (N\u2009=\u2009CCH3N), 9.50 (-CH2N-CCH3\u2009=\u2009C), 9.12 (H3CN-CCH3\u2009=\u2009C). HRMS , m/z calcd. for [2M \u2013 PF6]+ 959.3240, found 959.3247; m/z calcd. for [M \u2013 PF6]+ 407.1799, found 407.1828; m/z calcd. for [M \u2013 2PF6]2+ 131.1078, found 131.1071.To an aqueous solution of [How to cite this article: Shi, G. et al. Halides with Fifteen Aliphatic C-H\u00b7\u00b7\u00b7Anion Interaction Sites. Sci. Rep.6, 30123; doi: 10.1038/srep30123 (2016)."} +{"text": "III(N**)3] . Surprisingly, complex 1 exhibits a trigonal planar geometry in the solid state, which is unprecedented for three-coordinate actinide complexes that have exclusively adopted trigonal pyramidal geometries to date. The characterization data for [UIII(N**)3] were compared with the prototypical trigonal pyramidal uranium(III) triamide complex [UIII(N\u201c)3] (N\u201d=N(SiMe3)2\u2212), and taken together with theoretical calculations it was concluded that pyramidalization results in net stabilization for [UIII(N\u201c)3], but this can be overcome with very sterically demanding ligands, such as N**. The planarity of 1 leads to favorable magnetic dynamics, which may be considered in the future design of UIII single-molecule magnets.We report the synthesis and characterization of the uranium(III) triamide complex [U Both covalent[19] and electrostaticIII(N\u201c)3],III(N**)3] 2\u2212), which adopts an unprecedented trigonal planar geometry for an actinide triamide complex. Complex 1 is closely related to [UIII(N\u201d)3], allowing the contributions to pyramidalization to be assessed, together with the impact of geometry on magnetic (including dynamic) and electronic properties of UIII complexes, for the future rational design of useful An materials.f-Block metal centers favor high CNs, because Ln and An cations have relatively large ionic radii and bonding regimes that are dominated by electrostatic contributions.1 was prepared by a modification of the revised synthesis of [UIII(N\u201c)3].III(I)3(THF)4]2tBu)2}]2 in THF, followed by work-up and recrystallization from hexane to give 1 as dark purple needles in 62\u2009% yield (1 at \u22121 are attributed to the UNSi2 stretching modes of the silylamide ligand. The asymmetric stretch (950\u2005cm\u22121) is 40\u2005cm\u22121 lower than that observed for [UIII(N\u201d)3] (990\u2005cm\u22121),3] MNSi2 asymmetric stretches (ca. 50\u2005cm\u22121).6aComplex \u2009% yield .21 Absor1H\u2005NMR spectrum of 1 exhibits two resonances at \u03b4=3.8 (\u03bd\u00bd=206\u2005Hz) and \u221247.0\u2005ppm (\u03bd\u00bd=4597\u2005Hz) in a 54:36 ratio that are assigned to the tBuSi and Me2Si protons, respectively. The Me2Si resonance of 1 is much broader than the analogous resonance for [UIII(N\u201c)3] ,\u03b4=\u221232.9\u2005ppm, \u03bd\u00bd=266\u2005Hz).13C\u2005NMR spectrum of 1 exhibited two resonances for the Me2Si (\u03b4=\u22122.1 and 1.5\u2005ppm) and tBuSi quaternary carbons (\u03b4=18.2 and 32.0\u2005ppm), but only one for the tBuSi primary carbons (\u03b4=26.4\u2005ppm). In contrast, in the 13C\u2005NMR spectrum of [UIII{N(SiPhMe2)2}3], the Me2Si group resonates at \u03b4=\u221257.1\u2005ppm.29Si\u2005NMR spectrum of 1 at \u03b4 \u2212296.0\u2005ppm (\u03bd\u00bd=73\u2005Hz), which has not been reported for similar systems,III complex.23The 13\u21925f26d1 transitions at 20\u2009000 (\u03b5=776\u2009m\u22121\u2009cm\u22121) and 22\u2009500\u2005cm\u22121 (\u03b5=770\u2009m\u22121\u2009cm\u22121) that are typical of UIII[24] and comparable to a broad absorption observed for [UIII{N(SiPhMe2)2}3] at 21\u2009500\u2005cm\u22121 (\u03b5=430\u2009m\u22121\u2009cm\u22121).\u22121 region, weak Laporte forbidden 5f\u21925f transitions were observed (\u03b5=15\u201364\u2009m\u22121\u2009cm\u22121).III complexes, such as [U(I)3(THF)4]III{N(SiPhMe2)2}3],[27]The electronic absorption spectrum of 1 was determined and is depicted in Figure 1 crystallizes in the C2/c space group, with a twofold axis bisecting the U(1)\u2013N(1) bond. This contrasts to [Fe(N\u201c)3],III(N\u201d)3],III(N\u201c)3],III(N\u201d)3],P31c space group, and [UIII{N(SiPhMe2)2}3], which crystallizes in R3.1 is almost ideally trigonal planar, with U\u2013N bonds that are statistically identical within experimental uncertainty [U-N range 2.403(3)\u20132.415(6)\u2005\u00c5]. These distances are longer than those observed in [UIII(N\u201c)3] [2.320(4)\u2005\u00c5]III{N(SiPhMe2)2}3] [2.34(2)\u2005\u00c5],1 arising from the sterically demanding tBu groups. The U centroid/N(1)-N(2)-N(2A) mean plane distance in 1 is 0.008(2)\u2005\u00c5, and the N-U-N bond angles (range 119.1(2)\u2013120.47(9)\u00b0) sum to 360\u00b0; in contrast, [UIII(N\u201d)3] and [UIII{N(SiPhMe2)2}3] exhibit U centroids 0.456(1) and 0.874\u2005\u00c5 from the N3 planes, and the N-U-N angles average 116.24(7) (\u03a3 angles 348.72(7)\u00b0) and 106.88\u00b0 (\u03a3 angles 320.64\u00b0), respectively.2 fragments of 1 are essentially planar and all bisect the UN3 plane (range 53.23\u201361.35\u00b0) to form a molecular propeller.The crystal structure of III(N\u201c)3] and [UIII{N(SiPhMe2)2}3] are predicted by the polarized-ion model, whereby net stabilization was achieved by dipole formation.III(N\u201d)3] exhibits unequal U-N-Si angles (108.50(7) and 125.25(7)\u00b0), because one Si\u2013C bond for each N\u201c ligand is relatively close to the U center .\u22c5\u22c5\u22c5Si\u2013C\u03b3 interactions, as have been discussed for [UIII{CH(SiMe3)2}3]III(N\u201d)3].[31] The shortest U\u22c5\u22c5\u22c5C\u03b3 and U\u22c5\u22c5\u22c5Si distances in 1 are 3.119\u20133.301\u2005\u00c5 and 3.433\u20133.510\u2005\u00c5, respectively, and they are not correctly orientated to interact with the U center. Although there is no evidence for agostic U\u22c5\u22c5\u22c5Si\u2013C\u03b3 interactions in 1, stabilizing U\u22c5\u22c5\u22c5C\u2013H contacts cannot be discounted.The pyramidal geometries of [U1 and [UIII(N\u201c)3].1 , providing qualitative models 3] 0.393\u2005\u00c5). In both models, the HOMO, HOMO\u22121 and HOMO\u22122 represent the three unpaired UIII 5f electrons 3] 86.81, 86.32, 84.17\u2009% U 5f, respectively). Both models exhibit essentially insignificant degrees of U 6d/5f orbital contributions to the U\u2013N bonds, with the HOMO\u22123, HOMO\u22124. and HOMO\u22125, representing the \u03c0 components 3] 4.29/0, 0/2.06, 1.63/1.39\u2009% U 5f/6d, respectively) and the HOMO\u22126, HOMO\u22127, and HOMO\u22128 the \u03c3 components 3] 0/5.04, 0/5.26, 2.14/0\u2009% U 5f/6d, respectively). This concurs with gas-phase photoelectron spectroscopy (PES) studies of [U(N\u201d)3], which have shown that \u03c0 bonding between the ligand and U center is insignificant in this complex.1=\u22123.26; [UIII(N\u2032\u2032)3]=\u22123.26) are identical, which also supports similar bonding patterns for 1 and [UIII(N\u201c)3].Unrestricted DFT calculations were carried out on full models of III(CH3)3] III(NH2)3] 1 and [UIII(N\u201c)3] together with the small U 6d/5f contributions to the U\u2013N \u03c3 and \u03c0 components, we suggest that the experimentally determined trigonal planar geometry of 1 results from steric interactions involving the large N** ligands. These interactions could predominate over crystal packing forces, which are often only approximately 10\u2005kJ\u2009mol\u22121.1 and [UIII(N\u201d)3], therefore, the planar geometry of 1 derives principally from steric effects involving the ligands.Ab initio calculations on [An1 was calculated to be 2.59\u2005\u03bcB in [D6]benzene at 298\u2005K by using the Evans method.[36] Magnetometry measurements on a powdered sample of 1 suspended in eicosane gave a magnetic susceptibility temperature product, \u03c7T, of 1.07\u2005cm3\u2009Kmol\u22121 (2.92\u2005\u03bcB) at 298\u2005K,3 4I9/2 ground state (3.69\u2005\u03bcB), because not all crystal field levels are thermally occupied,III complexes described in the literature (range 2.13\u20134.63\u2005\u03bcB).[38] The \u03c7T value of 1 decreases to 0.41\u2005cm3\u2009Kmol\u22121 at 2\u2005K; ac measurements give a low-temperature plateau in the in-phase \u03c7\u2032T at 0.48\u2005cm3\u2009Kmol\u22121[21] consistent with thermal depopulation into a Kramers doublet ground state.1 are consistent with UIII,[27] and simulation gives geff=3.55, 2.97, and 0.553 for the ground Kramers doublet 3] is an SMM,1 to probe differences in the dynamic magnetic behavior as a result of the higher symmetry. Compound 1 is also an SMM, with clear frequency-dependent behavior 3], and maxima in the out-of-phase susceptibility \u03c7\u2032\u2032(T) are seen to significantly higher temperatures for 1 than for [UIII(N\u201c)3] at equivalent frequencies . An Arrhenius treatmentUeff=21.4\u00b10.2\u2005K for 1. Although this is lower than that reported for [UIII(N\u201d)3] (31\u2005K), the latter value was derived from an extremely limited temperature range\u03c4) at 2\u2005K is 2.6\u2005ms for 1; from the previously reported dataIII(N\u201c)3] at 2\u2005K, an order of magnitude quicker. The pre-factor \u03c40 for 1 is greater by four orders of magnitude (3.1\u00d710\u22127 cf. 10\u221211\u2005s for [UIII(N\u201d)3]).\u03c7\u2032 and \u03c7\u201c at 1.8\u2005K for 1\u03b1=0.001\u20130.03 from Cole\u2013Cole analysis), an order of magnitude lower than in [UIII(N\u201d)3] (\u03b1=0.09\u20130.34).1 at 1.8\u2005K on a conventional superconducting quantum interference device (SQUID) magnetometer 3].Compound from optical studies.1 and [UIII(N\u201d)3] are SMMs despite their easy-plane anisotropy: this highlights the complexity of interpreting f-block relaxation data,1 compared with [UIII(N\u201c)3] on flattening the geometry is because of quenched mixing. In Dh3 |zJ|=1/2 cannot mix with any other doublet within the 4I9/2 term, whereas in Cv3, it can mix with both |zJ|=5/2 and 7/2.In the trigonal planar geometry of 1 and [UIII(N\u201c)3] can be attributed to differences in symmetry that may be useful to consider in the future design of UIII SMMs with greater relaxation times. Computational analyses of 1 and [UIII(N\u201d)3] have shown only minor differences in their calculated bonding schemes, therefore, the energy gained by pyramidalization, which leads to favorable agostic M\u22c5\u22c5\u22c5Si\u2013C\u03b3 interactions in [UIII(N\u201c)3],To conclude, we have prepared and fully characterized an unprecedented trigonal planar actinide triamide complex. Differences in the spectroscopic and magnetic data between Synthesis of 1: THF (20\u2005mL) was added to a precooled (\u221278\u2009\u00b0C) mixture of [K{N(SiMe2tBu)2}]2 and [U(I)3(THF)4] . The reaction mixture was allowed to warm to RT slowly with stirring over 48\u2005h, with precipitation of a pale solid. Volatiles were removed in vacuo, and the dark purple solid was extracted with hexanes (3\u00d710\u2005mL). Recrystallization from hexanes (5\u2005mL) at \u221230\u2009\u00b0C gave 1 as dark purple needles .1H\u2005NMR : \u03b4=\u221247.04 2), 3.79\u2005ppm 3); 13C{1H}\u2005NMR : \u03b4=\u22122.13 (Si(CH3)2), 1.45 (Si(CH3)2), 18.22 (SiC(CH3)3), 26.40 (SiC(CH3)3), 31.98\u2005ppm (SiC(CH3)3); 29Si{1H}\u2005NMR : \u03b4=\u2212296.04\u2005ppm ; FTIR (Nujol); 2), 825 , 761 , 655 (m), 604 (s) cm\u22121; \u03bceff=2.59\u2005\u03bcB (Evans method); elemental analysis calcd for C36H90Si6N3U (971.67\u2005g\u2009mol\u22121): C 44.5, H 9.34, N 4.33; found: C 38.29, H 9.10, N 4.22. Low carbon values were obtained upon repeating the analysis multiple times on different batches and is ascribed to 1 being a silicon-rich molecule, as was observed previously.42"} +{"text": "H-1,2,4-triazol-3-yl-\u03baN4)acetato-\u03baO]di\u00adaqua)\u00adnickel(II) dihydrate, is the first transition metal complex of 2-acetic acid (ATAA).The title compound, bis\u00ad[(5-amino-1 4H5N4O2)2(H2O)2]\u00b72H2O, represents the first transition metal complex of the novel chelating triazole ligand, 2-acetic acid (ATAA), to be structurally characterized. In the mol\u00adecule of the title complex, the nickel(II) cation is located on an inversion centre and is coordinated by two water mol\u00adecules in axial positions and two O and two N atoms from two trans-oriented chelating anions of the deprotonated ATAA ligand, forming a slightly distorted octa\u00adhedron. The trans angles of the octa\u00adhedron are all 180\u00b0 due to the inversion symmetry of the mol\u00adecule. The cis-angles are in the range 87.25\u2005(8)\u201392.75\u2005(8)\u00b0. The six-membered chelate ring adopts a slightly twisted boat conformation with puckering parameters Q = 0.542\u2005(2)\u2005\u00c5, \u0398 = 88.5\u2005(2) and \u03d5 = 15.4\u2005(3)\u00b0. The mol\u00adecular conformation is stabilized by intra\u00admolecular N\u2014H\u22efO hydrogen bonds between the amino group and the chelating carboxyl\u00adate O atom of two trans-oriented ligands. In the crystal, the complex mol\u00adecules and lattice water mol\u00adecules are linked into a three-dimensional framework by an extensive network of N\u2014H\u22efO, O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds.The title compound, [Ni(C C-amino-1,2,4-triazoles are employed as polydentate ligands for the synthesis of coordination compounds with various metals that demonstrate useful spectroscopic, magnetic, biological and catalytic properties can act as chelating polydentate ligands acetic acid 2(H2O)2]\u00b72H2O (1).In a continuation of our work on the synthesis and reactivity of amino\u00adtriazole carb\u00adoxy\u00adlic acids \u201392.75\u2005(8)\u00b0. The third water mol\u00adecule is not involved in coordination. The anions of ATAA coordinate the NiII cation through the nitro\u00adgen atom N1 of the triazole ring and the oxygen atom O53 of the carboxyl\u00adate group \u2005\u00c5, \u0398 = 88.5\u2005(2), \u03d5 = 15.4\u2005(3)\u00b0. The Ni\u2014N1 bond length is 2.051\u2005(2)\u2005\u00c5, and the Ni\u2014O1 and Ni\u2014O53 bond lengths are 2.083\u2005(2) and 2.059\u2005(2)\u2005\u00c5, respectively, within the normal ranges for other reported NiII complexes , the Nion Fig.\u00a02. The traup Fig.\u00a02, similards Fig.\u00a02.In the crystal, mol\u00adecules of the complex and lattice water mol\u00adecules are linked into a three\u2013dimensional framework by extensive N\u2014H\u22efO, O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds Table\u00a01.et al., 2010H-1,2,4-triazole-3-carb\u00adoxy\u00adlic acid (ATCA) with various metals acetate (2) in an aqueous solution of nickel nitrate 2\u00b76H2O in water (5\u2005ml). After standing at room temperature for two weeks, the formed crystals were collected by filtration yielding the target compound (1).All attempts to prepare crystals of complex (1) suitable for X-ray investigation by mixing solutions of te Fig.\u00a04. A solut2 group and refined as riding, with Uiso(H) = 1.2Ueq(C). The N,O-bound H atoms that are involved in hydrogen bonds were found from difference Fourier maps. Their distances to the parent atoms were refined to be equal, with a common Uiso(H) value for pairs of related H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814021436/wm5066sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814021436/wm5066Isup2.hklStructure factors: contains datablock(s) I. DOI: 1026535CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Each AgI ion is bridged by the ligands, forming a helical chain propagating along the b-axis direction. The right- and left-handed helical chains are alternately arranged via Ag\u22efAg [3.2639\u2005(5)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.523\u2005(1)\u2005\u00c5], resulting in the formation of a two-dimensional supra\u00admolecular network extending parallel to (101). Weak Ag\u22efF inter\u00adactions [longest Ag\u22efF inter\u00adaction = 3.153\u2005(2)\u2005\u00c5], as well as N\u2014H\u22efF and C\u2014H\u22efF hydrogen-bonding inter\u00adactions, occur between the helical chains and the anions.In the title polymeric complex, {[Ag(C I coordination polymers with symmetrical dipyridyl ligands, see: Lee et al. ]PF6 = 0.027wR(F2) = 0.071S = 1.052740 reflections199 parametersH-atom parameters constrainedmax = 0.86 e \u00c5\u22123\u0394\u03c1min = \u22120.44 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, New_Global_Publ_Block. DOI: 10.1107/S1600536814011465/sj5402Isup2.hklStructure factors: contains datablock(s) I. DOI: 1003661CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The coordinating water mol\u00adecules are hydrogen bonded to the carboxyl O atoms [O \u22ef O = 2.6230\u2005(17)\u2005\u00c5], enclosing an S(6) hydrogen-bonding motif, while inter\u00admolecular O\u2014H\u22efO hydrogen bonds link two of the non-coordinating water mol\u00adecules to the coordinating water mol\u00adecules and NA anions. In the crystal, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules, enclosing The asymmetric unit of the monomeric cobalt complex, [Co(C 11H13O2)2(C6H6N2O)2(H2O)2]\u00b72H2O, contains one half of the complex mol\u00adecule, one coordinating and one non-coordinating water mol\u00adecule, one 4-tert-butyl\u00adbenzoate (TBB) ligand and one nicotinamide (NA) ligand; the Co atom lies on an inversion centre. All ligands coordinating to the Co atom are monodentate. The four nearest O atoms around the Co atom form a slightly distorted square-planar arrangement, with the distorted octa\u00adhedral coordination completed by the two pyridine N atoms of the NA ligands at distances of 2.1638\u2005(11)\u2005\u00c5. The coordinating water mol\u00adecules are hydrogen bonded to the carboxyl O atoms [O \u22ef O = 2.6230\u2005(17)\u2005\u00c5], enclosing an S(6) hydrogen-bonding motif, while inter\u00admolecular O\u2014H\u22efO hydrogen bonds link two of the non-coordinating water mol\u00adecules to the coordinating water mol\u00adecules and NA anions. The dihedral angle between the planar carboxyl\u00adate group and the adjacent benzene ring is 29.09\u2005(10)\u00b0, while the benzene and pyridine rings are oriented at a dihedral angle of 88.53\u2005(4)\u00b0. In the crystal, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules, enclosing R22(8), R22(10) and R44(12) ring motifs, forming layers parallel to (001). The C and H atoms of the tert-butyl group of the TBB ligand are disordered over two sets of sites with an occupancy ratio of 0.631\u2005(5):0.369\u2005(5). The asymmetric unit of the mononuclear cobalt complex, [Co(C The Co\u2014O bond lengths are 2.1104\u2005(11)\u2005\u00c5 (for water oxygens) and 2.1252\u2005(9)\u2005\u00c5 (for benzoate oxygens) and the Co\u2014N bond length is 2.1638\u2005(11)\u2005\u00c5, close to standard values. The Co1\u2014O2\u2014C1\u2014C2 torsion angle [\u2212163.00\u2005(9)\u00b0] causes a slight downward tilt of the ligand.The dihedral angle between the planar carboxyl\u00adate group (O1/O2/C1) and the adjacent benzene (C2\u2013C7) ring is 29.09\u2005(10)\u00b0, while the benzene and pyridine (N1/C9\u2013C13) rings are oriented at a dihedral angle of 88.53\u2005(4)\u00b0.w\u22efOc hydrogen bonds (Table\u00a01S(6) hydrogen-bonding motifs, while inter\u00admolecular O\u2014Hw\u22efOw and O\u2014Hw\u22efOna (na = nicotinamide) hydrogen bonds link two of the non-coordinating water mol\u00adecules to the coordinating water mol\u00adecules and NA anions in water (75\u2005ml) and nicotinamide in water (25\u2005ml) with sodium 4-tert-butyl\u00adbenzoate in water (250\u2005ml). The mixture was filtered and set aside to crystallize at ambient temperature for five days, giving pink single crystals.The title compound was prepared by the reaction of CoSO2), H41, H42, H51 and H52 (for H2O) were located in a difference Fourier map and were refined freely. The C-bound H atoms were positioned geometrically, with C\u2014H = 0.93 and 0.96\u2005\u00c5 for aromatic and methyl H atoms, respectively, and constrained to ride on their parent atoms, with Uiso(H) = k \u00d7 Ueq(C), where k = 1.5 for methyl H atoms and k = 1.2 for aromatic H atoms. During the refinement process the disordered t-butyl group atoms were refined with major:minor occupancy ratios of 0.631\u2005(5):0.369\u2005(5).Experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0210.1107/S2056989016008689/pk2579sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016008689/pk2579Isup2.hklStructure factors: contains datablock(s) I. DOI: 1482507CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "One of these P atoms belongs to a bridging dppb ligand [Cu\u2014P = 2.2381\u2005(5)\u2005\u00c5] and two belong to a chelating dppb ligand [Cu\u2014P = 2.2450\u2005(6) and 2.2628\u2005(5)\u2005\u00c5]. The bridging dppb ligand lies on an inversion centre. In the crystal, the cation and the PF6\u2212 anion are linked by C\u2014H\u22efF inter\u00adactions, forming a tape along [110]. The cation and the diethyl ether solvent mol\u00adecule are also linked by a C\u2014H\u22efO inter\u00adaction.In the centrosymmetric dinuclear copper(I) complex cation of the title compound, [Cu For cop al. 1999; Kitagaw al. 1995. For our al. 2006.2(C28H28P2)3](PF6)2\u00b72C4H10O = 0.036wR(F2) = 0.090S = 1.069436 reflections523 parametersH-atom parameters constrainedmax = 0.41 e \u00c5\u22123\u0394\u03c1min = \u22120.35 e \u00c5\u22123\u0394\u03c1CrystalClear used to solve structure: SIR92 global, I. DOI: 10.1107/S1600536814009763/is5355Isup2.hklStructure factors: contains datablock(s) I. DOI: 1000315CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Two copper(II) complexes, a dinuclear and a hexa\u00adnuclear complex with bridging hydroxyl and nitrate ligands, were obtained from reaction of copper nitrate with di\u00adhydroxy\u00adbipyridine at neutral and slightly acidic pH. Formation of multi-nuclear complexes contrasts with the equivalent sulfate compounds which formed discrete mononuclear complexes. The complexes feature intra\u00admolecular and inter\u00admolecular hydrogen bonding. 3)2 and 6,6\u2032-di\u00adhydroxy\u00adbipyridine (dhbp) exhibit bridging nitrate and hydroxide ligands. The dinuclear complex (\u03bc-hydroxido-1:2\u03ba2O:O\u2032)(\u03bc-nitrato-1:2\u03ba2O:O\u2032)(nitrato-1\u03baO)dicopper(II), [Cu2(C10H7N2O2)(OH)(NO3)2(C10H8N2O2)] or [Cu(6-OH-6\u2032-O-bpy)(NO3)(\u03bc-OH)(\u03bc-NO3)Cu], (I), with a 2:1 ratio of nitrate to hydroxide anions and one partially deprotonated dhbp ligand, forms from a water\u2013ethanol mixture at neutral pH. The hexa\u00adnuclear complex bis\u00adtetra\u00adkis\u00adtetra\u00adkis\u00ad(\u03bc-hydroxido-\u03ba2O:O\u2032)bis\u00ad(methanol-\u03baO)tetra\u00adkis\u00ad(\u03bc-nitrato-\u03ba2O:O\u2032)hexa\u00adcopper(II), [Cu6(C10H6N2O2)2(CH4O)2(OH)4(NO3)4(C10H8N2O2)4] or [Cu(\u03bc-NO3)2(\u03bc-OH)Cu(\u03bc-OH)Cu(CH3OH)]2, (II), with a 1:1 NO3\u2013OH ratio and two fully protonated and fully deprotonated dhbp ligands, was obtained by methanol recrystallization of material obtained at pH 3. Complex (II) lies across an inversion center. Complexes (I) and (II) both display intra\u00admolecular O\u2014H\u22efO hydrogen bonding. Inter\u00admolecular O\u2014H\u22efO hydrogen bonding links symmetry-related mol\u00adecules forming chains along [100] for complex (I) with \u03c0-stacking along [010] and [001]. Complex (II) forms inter\u00admolecular O\u2014H\u22efO hydrogen-bonded chains along [010] with \u03c0-stacking along [100] and [001].Two multinuclear complexes synthesized from Cu(NO A new coordination mode is observed for the mono-deprotonated (6-O-6\u2032-OH-bpy) ligand in which it bridges two metals, through coordination of N1,N2 to Cu1 and through bridging to Cu2 via the pyridino\u00adlate oxygen O2 [1.946\u2005(3)\u2005\u00c5]. The C\u2014O bond lengths (\u00c5) for the dhbp ligands are 1.335\u2005(5) (C1\u2014O1), 1.322\u2005(5) (C11\u2014O3), and 1.316\u2005(4) (C20\u2014O4) for the protonated hydroxyl groups and slightly shorter at 1.310\u2005(4) (C10\u2014O2) for the pyridino\u00adlate, reflecting double-bond character. Both copper atoms have a distorted square-pyramidal geometry with \u03c4 = 0.394 at Cu1 and \u03c4 = 0.119 at Cu2 \u2005\u00c5 hydrogen bond from O5 to O11 (hydroxyl to nitrate) with a bond angle of 164\u2005(4)\u00b0. Numerical details of the hydrogen bonds are given in Table\u00a01The dinuclear copper(II) dhbp complex (I)et al., 1984N,N bound), one bridging hydroxide to Cu2 (O14), one bridging nitrate to Cu2 (O11), and one bridging nitrate (O7) which tethers the two asymmetric units. The coordination of Cu2 entails one deprotonated dhbp , one bridg\u00ading nitrate to Cu1 (O12), and two bridging hydroxides to Cu1 and Cu3 . The remaining coord\u00adination sphere of Cu3 entails one dhbp , one methanol (O15), one bridging hydroxide (O13), and one bridging nitrate (O8) which tethers the two asymmetric units. Each deprotonated oxygen of dhbp acts as an acceptor for intra\u00admolecular hydrogen bonds from the two protonated dhbp ligands, O2 to O3 at 2.499\u2005(3)\u2005\u00c5 (O\u2014H\u22efO bond angle of 172\u00b0), and O6 to O4 at 2.495\u2005(3)\u2005\u00c5 (O\u2014H\u22efO bond angle of 168\u00b0) shown in Fig.\u00a04The hexa\u00adnuclear copper(II)Comparison of complex (I)II ions is charge balanced with one terminal and one bridging nitrate each with a single negative charge, one deprotonated hydroxyl group of dhbp, and one bridging hydroxide. The bond lengths to the bridging hydroxide from Cu1 to O5 is 1.964\u2005(3)\u2005\u00c5 and Cu2 to O5 is 1.939\u2005(3)\u2005\u00c5 where the proton of O5 hydrogen bonds to one acceptor. A comparable bond length is 1.946\u2005(3)\u2005\u00c5 from Cu2 to O2 of the deprotonated dhbp ligand. The remaining oxygen atoms of the dhbp ligands are each protonated and engaged in hydrogen bonding as described above. The asymmetric unit of complex (II)The bridging oxygen species for both complexes (I)vide supra. The dinuclear complex (I)x, \u2212y, \u2212z at a distance of 3.894\u2005(3)\u2005\u00c5 between the centroids of the rings. The pyridine ring containing N1 inter\u00adacts with its symmetry-equivalent ring across the symmetry operation 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, \u2212z with a centroid-to-centroid distance of 3.969\u2005(3)\u2005\u00c5. The dhbp ligand coordinating to Cu2 also shows \u03c0-stacking via two alternating inversion-symmetry operations, forming chains along [100]. The pyridine rings containing N3 and N4 inter\u00adcross by the symmetry operation 1\u00a0\u2212\u00a0x, \u2212y, 1\u00a0\u2212\u00a0z where the centroid of the ring defined by N3 is at a distance of 3.604\u2005(2)\u2005\u00c5 from the centroid of the ring defined as N4 on side of the dhbp plane with the bridging hydroxide. These rings also \u03c0-stack on the opposite face of the plane at a distance of 3.768\u2005(2)\u2005\u00c5 from the centroid of the ring defined by N3 to N4 through the symmetry operation \u2212x, \u2212y, 1\u00a0\u2212\u00a0z. Inter\u00admolecular hydrogen bonding from the bridging hydroxide ligand to the terminal oxygen of the bridging nitrate ligand inter\u00adlinks neighboring mol\u00adecules primarily along [100]. See Fig.\u00a06Some inter\u00admolecular hydrogen-bonding inter\u00adactions in both complexes have already been discussed, ac plane and exhibit \u03c0-stacking but in a less regular fashion than for complex (I)x, \u2212y, \u2212z with a centroid-to-centroid distance of 3.784\u2005(2)\u2005\u00c5 of the pyridine rings containing N1 to the ring containing N2 and vice versa. A single ring of each dhbp ligand bound to Cu2 and Cu3 \u03c0-stack via translation at a distance of 3.551\u2005(2)\u2005\u00c5 between the centroids of the pyridine rings defined by N3 and N5, respectively. Additionally, the Cu3 dhbp ligand \u03c0-stacks via the symmetry-equivalent ring defined by N5 of a neighboring mol\u00adecule across the symmetry operation \u2212x, \u2212y, 1\u00a0\u2212\u00a0z at a centroid-to-centroid distance of 3.887\u2005(2)\u2005\u00c5. Close proximity occurs in plane between the pyridine ring containing N4 of the dhbp ligand bound to Cu2 at a distance of 3.818\u2005(2)\u2005\u00c5 between C18 to C19 and vice versa across the symmetry operation 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, \u2212z.The hexa\u00adnuclear complex (II)et al., 2007et al., 2013et al., 2009et al., 2013et al., 2006et al., 2006et al., 2002Although many structures have been reported featuring a hydroxide anion bridging two copper(II) ions each bound by 2,2\u2032-bi\u00adpyridine, no analogous structure has been reported with a 6,6\u2032-dihy\u00addroxy-2,2\u2032-bi\u00adpyridine ligand. A search of the Cambridge Structural Database Copper(II)The neutral copper hexanuclear complex (II) Copper(II)Crystal data, data collection, and structure refinement details are summarized in Table\u00a03Cell Now involve domain 2 only, mean I/\u03c3 8.2, 34813 data (9811 unique) involve 2 domains, mean I/sigma 9.5, 11 data (11 unique) involve 3 domains, mean I/\u03c3 8.7 and 4 data (2 unique) involve 4 domains, mean I/\u03c3 57.6 The exact correlation matrix as identified by the integration program was found to be 1.00336 0.02923 \u22120.02720, \u22120.01894 1.02272 \u22120.04903, 0.02520 0.05747 0.97055. The data were corrected for absorption using TWINABS arom = 0.95\u2005\u00c5 with Uiso(H) = 1.2Ueq(C), C\u2014Hmeth\u00adyl = 0.98\u2005\u00c5 with Uiso(H) = 1.5Ueq(C). O\u2014H were refined for complex (I)Uiso(HOH) were set to 1.5Ueq(O).C- and O-bound H atoms were placed in calculated positions and allowed to ride on their carrier atoms: aromatic C\u2014H10.1107/S205698901502037X/lh5780sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S205698901502037X/lh5780Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S205698901502037X/lh5780IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1433678, 1000450CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The coordination geometry of the metal ion is distorted trigonal bipyramidal, with the ligand O atoms in the axial sites. Two of the carboxyl\u00adate groups of the ligand remain protonated and form short symmetric O\u2014H\u22efO hydrogen bonds. In the crystal, the ribbons inter\u00adact via a network of O\u2014H\u22efO hydrogen bonds in which coordinating water mol\u00adecules act as donors and carboxyl\u00adate O atoms within adjacent ribbons act as acceptors, giving rise to a three-dimensional framework. O\u2014H\u22efN inter\u00adactions are also observed. The asymmetric unit contains quarter of the ligand and the complete ligand has 2/m symmetry; the Li+ ion lies on a special position with m.. site symmetry. Both bridging water mol\u00adecules have m2m site symmetry and both lattice water mol\u00adecules have m.. site symmetry; one of the latter was modelled with a site occupancy of 0.25.The title coordination polymer, {[Li For the structures of related lithium complexes with pyrazine-2,3,5,6-tetra\u00adcarboxyl\u00adate and water ligands, see: Starosta & Leciejewicz 2010, 2014 \u25b6.2(C8H2N2O8)(H2O)2]\u00b72.5H2O = 0.042wR(F2) = 0.122S = 1.071151 reflections85 parameters3 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.25 e \u00c5\u22123\u0394\u03c1min = \u22120.29 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: SHELXS97 I, New_Global_Publ_Block. DOI: 10.1107/S160053681401174X/hb7224Isup2.hklStructure factors: contains datablock(s) I. DOI: 1004410CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-(aryl\u00adsulfon\u00adyl)-4-fluoro\u00adbenzamides, namely 4-fluoro-N-(2-methyl\u00adphenyl\u00adsulfon\u00adyl)\u00adbenzamide, (I), N-(2-chloro\u00adphenyl\u00adsulfon\u00adyl)-4-fluoro\u00adbenzamide, (II), and N-(4-chloro\u00adphenyl\u00adsulfon\u00adyl)-4-fluoro\u00adbenzamide monohydrate, (III), are described and compared with related structures. The conformation of the three mol\u00adecules is very similar with the aromatic rings being inclined to one another by 82.83\u2005(11) and 85.01\u2005(10)\u00b0 in the two independent mol\u00adecules of (I), 89.91\u2005(10)\u00b0 in (II) and 81.82\u2005(11)\u00b0 in (III).The crystal structures of three N-aryl\u00adsulfonyl-4-fluoro\u00adbenzamides, namely 4-fluoro-N-(2-methyl\u00adphenyl\u00adsulfon\u00adyl)benzamide, C14H12FNO3S, (I), N-(2-chloro\u00adphenyl\u00adsulfon\u00adyl)-4-fluorobenzamide, C13H9ClFNO3S, (II), and N-(4-chloro\u00adphenyl\u00adsulfon\u00adyl)-4-fluoro\u00adbenzamide monohydrate, C13H9ClFNO3S\u00b7H2O, (III), are described and compared with related structures. The asymmetric unit of (I) contains two independent mol\u00adecules (A and B), while that of (II) contains just one mol\u00adecule, and that of (III) contains a mol\u00adecule of water in addition to one main mol\u00adecule. The dihedral angle between the benzene rings is 82.83\u2005(11)\u00b0 in mol\u00adecule A and 85.01\u2005(10)\u00b0 in mol\u00adecule B of (I), compared to 89.91\u2005(10)\u00b0 in (II) and 81.82\u2005(11)\u00b0 in (III). The crystal structure of (I) features strong N\u2014H\u22efO hydrogen bonds between the A and B mol\u00adecules, resulting in an R44(16) tetra\u00admeric unit. These tetra\u00admeric units are connected into sheets in the bc plane by various C\u2014H\u22efO inter\u00adactions, and adjacent sheets are further inter\u00adlinked via C\u2014H\u22ef\u03c0ar\u00adyl inter\u00adactions, forming a three-dimensional architecture. The crystal structure is further stabilized by \u03c0ar\u00adyl\u2013\u03c0ar\u00adyl and S=O\u22ef\u03c0ar\u00adyl inter\u00adactions. In the crystal of (II), mol\u00adecules are connected into R22(8) and R22(14) dimers via N\u2014H\u22efO hydrogen bonds and C\u2014H\u22efO inter\u00adactions, respectively; the dimers are further inter\u00adconnected via a weak C=O\u22ef\u03c0ar\u00adyl inter\u00adaction, leading to the formation of chains along [1-10]. In the crystal of (III), N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds involving both the main mol\u00adecule and the solvent water mol\u00adecule results in the formation of sheets parallel to the bc plane. The sheets are further connected by C\u2014H\u22efO inter\u00adactions and weak C\u2014Cl\u22ef\u03c0ar\u00adyl, C\u2014F\u22ef\u03c0ar\u00adyl and S=O\u22ef\u03c0ar\u00adyl inter\u00adactions, forming a three-dimensional architecture.The crystal structures of three An intra\u00admolecular C14B\u2013-H14B\u22efO2B hydrogen bond S(6) ring motif.The asymmetric unit of compound (I)B) Fig.\u00a01, that diB) Fig.\u00a01 containsB) Fig.\u00a01 crystallnt Fig.\u00a01. This isA\u2014H1A\u22efO1B and N1B\u2014H1B\u22efO1A hydrogen bonds , while in the second level the tetra\u00admeric unit has a graph-set motif of R44(16). Adjacent tetra\u00admers are connected into sheets in the bc plane ar\u00adyl\u2013\u03c0ar\u00adyl inter\u00adactions \u2005\u00c5; Cg1 and Cg2 are the centroids of the fluoro\u00adbenzoyl rings of mol\u00adecules A and B, respectively] and also by weak S1A=O2A\u22ef\u03c0ar\u00adyl inter\u00adactions .The crystal structure of (I)s Table\u00a01 between it Fig.\u00a04. The unine Fig.\u00a04 via C6A\u2014s Table\u00a01. AdjacenB) Fig.\u00a05 to form ns Fig.\u00a06 connect these dimers, thus forming a one-dimensional architecture .In the crystal of (II)re Fig.\u00a07b.via bridging water mol\u00adecules, through strong N1\u2014H1\u22efO4, O4\u2014H1O4\u22efO1, O4\u2014H2O4\u22efO2 and O4\u2014H2O4\u22efO3 hydrogen bonds \u2005\u00c5], C11\u2014F1\u22efCg2 [F1\u22efCg2 = 3.8674\u2005(17)\u2005\u00c5] and S1=O2\u22efCg1 inter\u00adactions [O2\u22efCg1 = 3.2039\u2005(17)\u2005\u00c5] , forming a complex three-dimensional architecture s Table\u00a03, resultie Figs. 8 and 9 \u25b8.viz N-(aryl\u00adsulfon\u00adyl)-4-(substituted)benzamides, gave 14 hits. These fourteen compounds along with the three title compounds, (I)\u2013(III), are grouped into three series; series 1: N-(2-methyl\u00adphenyl\u00adsulfon\u00adyl)benzamide, N-(2-methyl\u00adphen\u00adyl\u00adsulfon\u00adyl)-4-(chloro/meth\u00adyl/nitro/meth\u00adoxy)benzamides and (I)series 2: N-(2-chloro\u00adphenyl\u00adsulfon\u00adyl)benzamide, N-(2-chloro\u00adphenyl\u00adsulfon\u00adyl)-4-(chloro/meth\u00adyl/nitro/meth\u00adoxy)benzamides and (II)series 3: N-(4-chloro\u00adphenyl\u00adsulfon\u00adyl)benzamide, N-(4-chloro\u00adphenyl\u00adsulfon\u00adyl)-4-(chloro/meth\u00adyl/nitro)benzamides and (III)A search of the Cambridge Structural Database benzamide -4-nitro\u00adbenzamide -4-meth\u00adoxy\u00adbenzamide -4-chloro\u00adbenzamide -4-methyl\u00adbenzamide -4-fluoro\u00adbenzamide (I)ortho-methyl group on the benzene\u00adsulfonyl ring is syn to the N\u2014H bond in the central \u2013C\u2013SO2--N\u2013C(O)\u2013 segment. The values of the dihedral angle between the two aromatic rings in the mol\u00adecules of series 1 fall in the range 73.9\u2005(1)\u2013 89.4\u2005(1)\u00b0, the smallest dihedral angle being in N-(2-methyl\u00adphenyl\u00adsulfon\u00adyl)benzamide and the largest in N-(2-methyl\u00adphenyl\u00adsulfon\u00adyl)-4-chloro\u00adbenzamide hydrogen bonds. However, except for compound (I)1 Table\u00a04, the asye Table\u00a04. CompariSeries 2: The asymmetric units of all of the compounds in series 2 \u2013 segment. The values of the dihedral angle between the two aromatic rings in the mol\u00adecules fall in the range 73.3\u2005(1)\u201389.91\u2005(10)\u00b0, which is almost the same as in series 1, the smallest being in N-(2-chloro\u00adphenyl\u00adsulfon\u00adyl)benzamide -4-fluoro\u00adbenzamide (II)N-(2-chloro\u00adphenyl\u00adsulfon\u00adyl)-benzamide, N-(2-chloro\u00adphenyl\u00adsulfon\u00adyl)-4-chloro\u00adbenzamide -4-methyl\u00adbenzamide hydrogen bonds, while strong N\u2014H\u22efO(S) hydrogen bonds in N-(2-chloro\u00adphenyl\u00adsulfon\u00adyl)-4-nitro\u00adbenzamide -4-meth\u00adoxy\u00adbenzamide -4-fluoro\u00adbenzamide (II)via C=O\u22ef\u03c0ar\u00adyl inter\u00adactions, forming a one-dimensional architecture.2 Table\u00a05 contain Series 3: In series 3, the parent compound N-(4-chloro\u00adphenyl\u00adsulfon\u00adyl)benzamide -4-chloro\u00adbenzamide -4-methyl\u00adbenzamide -4-nitro\u00adbenzamide -4-fluoro\u00adbenzamide (III)N-(4-chloro\u00adphenyl\u00adsulfon\u00adyl)benzamide and the largest for N-(4-chloro\u00adphenyl\u00adsulfon\u00adyl)-4-methyl\u00adbenzamide (Table\u00a06e Table\u00a06. Except Compounds (I)\u2013(III) were prepared by refluxing a mixture of 4-fluoro\u00adbenzoic acid, the corresponding substituted benzene\u00adsulfonamides and phospho\u00adrousoxychloride for 3\u2005h on a water bath. The resultant mixtures were cooled and poured into ice-cold water. The solids obtained were filtered, washed thoroughly with water and then dissolved in sodium bicarbonate solutions. The compounds were later re-precipitated by acidifying the filtered solutions with dilute HCl. They were filtered, dried and recrystallized. [Melting point (m.p.) of (I)Uiso = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. In the final cycles of refinement, reflections (0 1 1), (0 0 2) and I, II, III, global. DOI: 1470505, 1470504, 1470503CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compounds, (I), (II) and (III), are indole derivatives. Compounds (I) and (II) present two independent moieties in the asymmetric unit, and their packing is led by C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions. In compound (III), the C\u2014H\u22efO hydrogen bonds form 17H13NO2S, (I), C17H13NO3S, (II), and C24H17ClN2O5S\u00b7CHCl3, (III), are indole derivatives. Compounds (I) and (II) crystalize with two independent mol\u00adecules in the asymmetric unit. The indole ring systems in all three structures deviate only slightly from planarity, with dihedral angles between the planes of the pyrrole and benzene rings spanning the tight range 0.20\u2005(9)\u20131.65\u2005(9)\u00b0. These indole ring systems, in turn, are almost orthogonal to the phenyl\u00adsulfonyl rings [range of dihedral angles between mean planes = 77.21\u2005(8)\u201389.26\u2005(8)\u00b0]. In the three compounds, the mol\u00adecular structure is stabilized by intra\u00admolecular C\u2014H\u22efO hydrogen bonds, generating S(6) ring motifs with the sulfone O atom. In compounds (I) and (II), the two independent mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, while in compound (III), the mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, generating R22(22) inversion dimers.The title compounds, C The corresponding bond lengths and bond angles of mol\u00adecules A and B [in compounds (I)A and B of compound (I)], 0.20\u2005(9) and 0.86\u2005(9) [mol\u00adecules A and B of compound (II)], and 1.34\u2005(14)\u00b0 [compound (III)].The mol\u00adecular structures of title compounds (I)The indole ring systems are almost orthogonal to the phenyl\u00adsulfonyl rings [dihedral angles = 77.21\u2005(8) and 89.26\u2005(8)\u00b0 in (I)sp2 bond lengths are longer than the mean value of 1.355\u2005(14)\u00c5 for the N\u2014C bond length ring motifs with the sulfone O atom (Tables 1The sums of the bond angles around atoms N1 are 351.55 and 356.16\u00b0 in (I) Tables 1 and 3 \u25b8.via inter\u00admolecular C16B\u2014H16B\u22efO2A hydrogen bonds running parallel to the [101] direction. The crystal packing is further stabilized by inter\u00admolecular C10A\u2014H10A\u22efCg1, C11A\u2014H11A\u22efCg2 and C16A\u2014H16A\u22efCg3 inter\u00adactions, with separations of 3.727\u2005(2), 3.546\u2005(2) and 3.699\u2005(3)\u2005\u00c5 at and , respectively. Cg2 is the centre of gravity of pyrrole ring N1B/C1B/C6B/C7B/C8B, and Cg1 and Cg3 are the centres of gravity of benzene rings C1B\u2013C6B and C1A\u2013C6A, respectively. C\u2014H\u22ef\u03c0 inter\u00adactions run parallel to the [210] direction n Table\u00a01.A and B) are linked by inter\u00admolecular C12B\u2014H12B\u22efO2A hydrogen bonds and are further connected by C5A\u2014H5A\u22efCg1 and C17B\u2014H17C\u22efCg2 inter\u00adactions, with separations of 3.550\u2005(2) and 3.729\u2005(2)\u2005\u00c5 at (1 and Cg2 are the centres of gravity of benzene rings C9A\u2013C14A and C1A\u2013C6A), respectively). The C12B\u2014H12B\u22efO2A and C17B\u2014H17C\u22efCg2 inter\u00adactions run parallel to the [101] direction and C5A\u2014H5A\u22efCg1 inter\u00adactions run along the tri\u00adphenyl\u00adphospho\u00adnium ylide in dry toluene (20\u2005ml) was refluxed for 12\u2005h under an N2 atmosphere. After consumption of the starting material [monitered by thin-layer chromatography (TLC)], removal of the solvent in vacuo followed by column chromatographic purification gave (I)A solution of tri\u00adphenyl\u00adphosphonium ylide and 5-chloro\u00adnitro\u00adbenzaldehyde in dry chloro\u00adform (50\u2005ml) was refluxed for 10\u2005h under an N2 atmosphere. Removal of the solvent in vacuo followed by titration of the crude product with methonal (10\u2005ml), gave (III)A solution of [(3-acetyl-1-phenyl\u00adsufanyl-1Uiso(H) = 1.5Ueq(methyl C) and 1.2Ueq(nonmethyl C). The rotation angles for methyl groups were optimized by least squares.Crystal data, data collection and structure refinement details for compounds (I)10.1107/S2056989015014917/bg2558sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989015014917/bg2558Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989015014917/bg2558IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989015014917/bg2558IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989015014917/bg2558IIsup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015014917/bg2558IIIsup6.cmlSupporting information file. DOI: 1417660, 1417659, 1417658CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the hydrogen phosphate anions are linked by O\u2014H\u22efO hydrogen bonds into chains parallel to [100]. The inorganic anionic chains and the organic cations are linked by N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, forming a two-dimensional supra\u00admolecular network extending parallel to (001). 6H9N2+\u00b7HPO42\u2212, comprises two 2-amino\u00adanilinium cations and one hydrogen phosphate dianion. In the crystal, the HPO42\u2212 dianions are linked by O\u2014H\u22efO hydrogen bonds into chains along [100]. The inorganic anionic chains and organic cations are linked by N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, forming a two-dimensional supra\u00admolecular network extending parallel to (001).The asymmetric unit of the title compound, 2C Organic mono\u00adhydrogen (HPO42\u2212) and di\u00adhydrogen phosphate (H2PO4\u2212) compounds provide a class of materials with numerous practical and potential uses in various fields such as biomolecular sciences, catalysis, liquid-crystal-material development, ferroelectrics, non-linear optical and supra\u00admolecular studies \u00b0].The asymmetric unit of the title compound comprises two 2-amino\u00adanilinium cations and one hydrogen phosphate dianion Fig.\u00a01. The exiviz. O\u2014H\u22efO, N\u2014H\u22efO and N\u2014H\u22efN (Table\u00a01D\u22efO2i hydrogen bond connects adjacent hydrogen phosphate anions, forming anionic chains extending along [100]. The oxygen atom O3 acts as a trifurcated hydrogen-bond acceptor for the donor nitro\u00adgen atom N1 at , and , forming a one-dimensional supra\u00admolecular ladder extending along [100] as shown in Fig.\u00a02C\u22efO3, N3\u2014H3A\u22efO3i and N3\u2014H3B\u22efO3iv hydrogen bonds, forming two types of fused rings of B\u22efO2, N1\u2014H1A\u22efO4ii and N1\u2014H1C\u22efO4iii hydrogen bonds, resulting in the formation of a two-dimensional organic\u2013inorganic supra\u00admolecular layered network parallel to (001) \u2212x, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a02; (v) x, \u22121\u00a0+\u00a0y, z] hydrogen bonds stabilize the (001) network. In the crystal structure Fig.\u00a03. In the re Fig.\u00a04, adjacenConQuest 1.17; Groom & Allen, 2014et al., 2002et al., 2012et al., 2010et al., 1998et al., 2005A CSD database search = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016004709/rz5186sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016004709/rz5186Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016004709/rz5186Isup3.cmlSupporting information file. DOI: 1469440CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two symmetry-related tetra\u00adbromo\u00adphthalate anions bridge the two CuII cations, forming a centrosymmetric dinuclear complex in which the Cu\u22efCu separation is 5.054\u2005(2)\u2005\u00c5. Intra\u00admolecular classic O\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22efO hydrogen bonds occur in the dinuclear mol\u00adecule. In the crystal, the mol\u00adecules are linked by weak C\u2014H\u22efBr and C\u2014H\u22efO inter\u00adactions into supra\u00admolecular chains propagating along the b-axis direction.In the title complex, [Cu For the al. 2004; Colacio al. 2009; Rodpun al. 2015; Yang et al. 2002. For the al. 2009; Kozlev\u010d al. 2004. For sup al. 2013; Stibran al. 2009; Aaker\u00f6y al. 2003; Vald\u00e9s- al. 1993; Julve e al. 1984. For mol al. 1984; Mendy e al. 2010.2(C8Br4O4)2(C6H16N2)2(H2O)2] = 0.016wR(F2) = 0.039S = 1.073739 reflections245 parameters2 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.80 e \u00c5\u22123\u0394\u03c1min = \u22120.41 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, New_Global_Publ_Block. DOI: 10.1107/S2056989015015194/xu5866Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015015194/xu5866fig1.tif. DOI: The structure with displacement ellipsoids drawn at the 30% probability level and H atoms shown as small spheres of arbitrary radii.Click here for additional data file.10.1107/S2056989015015194/xu5866fig2.tif. DOI: View of inter\u00adactions in the crystal.1418832CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I atoms in the dication of the title structure, [(C6H5)2PCH2P(C6H5)2Au2]Cl2\u00b7(CH3)2C=O\u00b7H2O, show an aurophilic inter\u00adaction of 2.9743\u2005(2)\u2005\u00c5.The Au 2{(C6H5)2PCH2P(C6H5)2}]Cl2\u00b7(CH3)2C=O\u00b7H2O, the dication forms an eight-membered {\u2014PCPAu}2 ring with a transannular aurophilic inter\u00adaction [Au\u22efAu = 2.9743\u2005(2)\u2005\u00c5]. The ring approximates a flattened boat conformation, with the two methyl\u00adene C atoms lying ca 0.58\u20130.59\u2005\u00c5 above the least-squares plane defined by the Au2P4 atoms (r.m.s. deviation = 0.0849\u2005\u00c5). One Cl\u2212 anion functions as a weak bridge between the AuI atoms [Au\u22efCl = 2.9492\u2005(13) and 2.9776\u2005(12)\u2005\u00c5]. The second Cl\u2212 anion forms two (water)O\u2014H\u22efCl hydrogen bonds about a centre of inversion, forming a centrosymmetric eight-membered {\u22efHOH\u22efCl}2 supra\u00admolecular square. Globally, the dications and loosely associated Cl\u2212 anions assemble into layers lying parallel to the ac plane, being connected by C\u2014H\u22efCl,\u03c0(phen\u00adyl) inter\u00adactions. The supra\u00admolecular squares and solvent acetone mol\u00adecules are sandwiched in the inter-layer region, being connected to the layers on either side by C\u2014H\u22efCl,O(acetone) inter\u00adactions.In the title complex salt, [Au These compounds are cytotoxic and kill cancer cells by initiating apoptotic pathways carbonimido\u00adthio\u00adates, 2C6H4} ]Cl2, was isolated as an acetone monosolvate monohydrate, (I)2(Ph2PCH2PPh2)]Cl2 salts characterized as an acetone solvate thiol\u00adates, reactions with the bipodal mol\u00adecule, {1,4-[MeOC(=S)N(H)]2(Ph2PCH2PPh2)]2+ dication, two Cl\u2212 anions, and a solvent mol\u00adecule each of acetone and water; all species are in general positions. The mol\u00adecular structure of the dication is shown in Fig.\u00a01I atoms are bridged by two Ph2PCH2PPh2 ligands, forming an eight-membered {\u2014PCPAu}2 ring. The ring has the form of a boat with the methyl\u00adene-C1 and C2 atoms lying to one side of the ring and 0.589\u2005(5) and 0.581\u2005(5)\u2005\u00c5, respectively, above the least-squares plane through the Au2P4 atoms which have a r.m.s. deviation of 0.0849\u2005\u00c5. There is a transannular Au1\u22efAu2 (aurophilic) inter\u00adaction of 2.9743\u2005(2)\u2005\u00c5. This inter\u00adaction is partly responsible for the deviations of the P1\u2014Au1\u2014P3 and P2\u2014Au2\u2014P4 angles from the ideal 180\u00b0, i.e. 173.24\u2005(4) and 170.04\u2005(4)\u00b0, respectively. The Au\u2014P bond lengths are almost equivalent, ranging from a short Au1\u2014P1 2.3061\u2005(12) to a long Au2\u2014P4 2.3130\u2005(12)\u2005\u00c5. The Cl1\u2212 anion forms a weak bridge between the two AuI atoms with Au1\u22efCl1 and Au2\u22efCl2 separations of 2.9492\u2005(13) and 2.9776\u2005(12)\u2005\u00c5, respectively. The second Cl\u2212 anion participates in hydrogen bonding as described below in Supra\u00admolecular features.The asymmetric unit of (I)2 supra\u00admolecular squares with edge lengths of 3.217\u2005(5) and 3.200\u2005(5)\u2005\u00c5, Table\u00a01ac plane, corresponding to the inter-layer region between layers of dications and Cl1\u2212 anions, Fig.\u00a02\u2212 anion forms a single (phen\u00adyl)C\u2014H\u22efCl contact, a reduced propensity reflecting its close association with the AuI atoms (see above). By contrast, the Cl2\u2212 anion forms four independent C\u2014H\u22efCl2 inter\u00adactions, i.e. a (methyl\u00adene)C\u2014H\u22efCl2 and three (phen\u00adyl)C\u2014H\u22efCl2 inter\u00adactions, providing links between the {\u22efHOH\u22efCl}2 rings and the cations. The acetone solvent mol\u00adecule accepts a (methyl\u00adene)- and a (phen\u00adyl)C\u2014H\u22efO contact.The most notable feature of the crystal packing of (I)2(Ph2PCH2PPh2)]Cl2 salt has been characterized twice previously, originally as an acetone solvate 2(Ph2PCH2PPh2)]Cl2 salts relates to the mode of association between the complex Au cations and Cl\u2212 anions. As noted above and shown in Fig.\u00a04a, the Cl1\u2212 anion in (I)I atoms. In the acetone solvate \u2005\u00c5. A similar pattern is noted in the aceto\u00adnitrile solvate ]Cl2 salts with loosely associated Cl\u2212 anions having greater distortions from linearity, in particular for the acetone solvate (AuCl)2 in aceto\u00adnitrile (50\u2005ml) was added NaOH and {1,4-[MeOC(=S)N(H)]2C6H4} in aceto\u00adnitrile (50\u2005ml). The resulting mixture was stirred at 323\u2005K for 2\u2005h. The final product was extracted with di\u00adchloro\u00admethane (100\u2005ml) and the solution was left for evaporation at room temperature. After 3 weeks a slurry formed. This was redissolved in a solvent mixture of acetone/aceto\u00adnitrile and left for slow evaporation. Colourless crystals were obtained after 10 days. Yield: 0.213\u2005g (43%). M.p. 477\u2013479\u2005K. 1H NMR : \u03b4 7.96 , 7.49 , 7.41 , 4.84 , 2.82 . Analysis calculated for C53H52Au2Cl2O2P4: C, 48.61; H, 4.00. Found: C, 48.64; H, 3.99. IR (cm\u22121): 3044 (m) \u03bd(C\u2014H), 1484 (s) \u03bd(C\u2014C).The title compound is an unexpected product from the reaction of bis\u00ad[chlorido\u00adgold(I)] bis\u00ad(di\u00adphenyl\u00adphosphane)methane with an equimolar amount of {1,4-[MeOC(=S)N(H)]Uiso(H) set to 1.2\u20131.5Uequiv(C). The water-bound H atoms were refined with O\u2014H = 0.84\u00b10.01\u2005\u00c5, and with Uiso(H) = 1.5Uequiv(O). The U33 parameter was elongated for the C93 atom. In the final refinement this was restrained to be nearly isotropic using the ISOR command in SHELXL I, global. DOI: 10.1107/S2056989015013341/wm5185Isup2.hklStructure factors: contains datablock(s) I. DOI: 1412185CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title platinum(II) complex shows a trigonal\u2013bipyramidal coordination and inter\u00admolecular C\u2014H\u22efCl, C\u2014H\u22ef\u03c0 and (C/O)\u2014H\u22efO hydrogen bonds. 11H13O2)Cl(C5H11N)]\u00b7C2H5OH, the PtII cation is penta\u00adcoordinated in a distorted square-planar geometry. In the crystal, inversion dimers showing C\u2014H\u22efCl and C\u2014H\u22ef\u03c0 inter\u00adactions are further stacked in columns along the a axis via C\u2014H\u22ef\u03c0 inter\u00adactions. The ethanol solvate mol\u00adecule inter\u00adacts with neighbouring meth\u00adoxy groups of methyl\u00adeugenol through O\u2014H\u22efO hydrogen bonds.In the title compound, [Pt(C In order to avoid steric hindrance between Cl1 and the two ring systems, especially atoms C2 and C12, both rings rotate along their bond with Pt1. This is easier for the piperidine ring [resulting in a C12\u2014N1\u2014Pt1\u2014Cl1 torsion angle of 70.7\u2005(2)\u00b0] than for the phenyl ring [C2\u2014C1\u2014Pt1\u2014Cl1 torsion angle of only \u221225.0\u2005(4)\u00b0]. As a consequence the H12B\u22efCl1 distance (2.831\u2005\u00c5) is larger than the H2\u22efCl1 distance (2.789\u2005\u00c5).In [PtCl(Meug-1H)(piperidine)], the Ptom Fig.\u00a01. The twoA\u22efCg1 inter\u00adactions, Cg1 is the centroid of the C1\u2013C6 aromatic ring, see Table\u00a01A\u22efCg1 inter\u00adactions. The ethanol mol\u00adecule inter\u00adacts by bifurcated O\u2014H\u22efO hydrogen bonds with both meth\u00adoxy groups of methyl\u00adeugenol and further on by C\u2014H\u22efO inter\u00adactions to a neighboring meth\u00adoxy group. No voids are present in the crystal packing.In the crystal packing Fig.\u00a02, the com3 torsion angles vary between \u221228 and +32\u00b0 (only 11 torsion angles are outside this region).The Cambridge Structural Database 2] was prepared from K[PtCl3(Meug)] in high yield (85%) according to Da et al. 2] and 6ml of acetone. The reaction mixture was stirred at room temperature for 30\u2005min. The obtained solution was cooled to 255\u2005K after which the precipitate was collected and washed with Et2O. The yield was 320\u2005mg (65%). The powder was dissolved in an acetone\u2013ethanol mixture. Colourless plate-like crystals were harvested after slow evaporation of acetone at room temperature.The title compound was synthesized by adding a solution of 1\u2005mmol of piperidine in 3\u2005ml of acetone to a mixture of 408\u2005mg (0.5\u2005mmol) of : C16H24ClNO2Pt, M = 491\u2013495 au; found : 490\u2013494 ([M\u2212H]+).IR with C\u2014H distances of 0.95 (aromatic), 0.98 (meth\u00adyl) and 0.99\u2005\u00c5 (methyl\u00adene), N\u2014H distance of 0.93 (NH) and O\u2014H distance of 0.84\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814023575/rz5134sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814023575/rz5134Isup2.hklStructure factors: contains datablock(s) I. DOI: 1031185CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the brightly green fluorescent Zn-complex of the hardly luminescent dipyrrin, the metal ion is bound by two dipyrrin ligands, as revealed first by its mass spectra. The crystal structure of this Zn-complex of the dipyrrin confirmed a regular 2:1 composition of the bidentate dipyrrin ligand and the metal ion. The latter was coordinated in a distorted tetrahedral fashion, as found in other dipyrrin Zn-complexes. The here studied Zn-complex of a designed dipyrrin ligand provides insights into the coordination properties of the proposed (2:1)- and (2:2)-complexes of phylloxanthobilin and bilirubin, respectively, which are two abundant natural bilin-type tetrapyrroles.A high yield preparation, spectroscopic and crystallographic investigation of the crystalline Zn-complex of a di( Dipyrrins (or dipyrromethenes) feature two conjugated pyrrole rings and represent dipyrrolic building blocks for the construction of porphyrins and related tetrapyrrolic macrocycles . The com\u03b2,\u03b2\u2032-sulfoleno)pyrrin 2.In the context of our recent interest in the metal coordination properties of yellow chlorophyll catabolites (phylloxanthobilins) , and of \u03b2,\u03b2\u2032-sulfoleno)pyrrin 2 relied on the earlier made corresponding dipyrromethane (1), available, in turn, from condensation of 3,5-di-tert-butylbenzaldehyde and \u03b2,\u03b2\u2032-sulfolenopyrrole + at m/z\u00a0=\u00a01087.2. Corresponding significant fragments occurred at m/z\u00a0=\u00a01023.3, 894.4, and 830.4, due to consecutive loss of the one, three, and four SO2-groups, respectively. The derived molecular formula of the Zn-complex 3 suggested the presence of two dipyrrin ligands 2 and one Zn(II)-ion, i.e. to represent Zn-(2)2 (see Scheme\u00a0The molecular formula of the dipyrrin Zn-complex 3 displayed a UV/Vis-spectrum in CH2Cl2 that featured a maximum at 487\u00a0nm (and a shoulder at 463\u00a0nm), corresponding to a 51\u00a0nm bathochromic shift, when compared with the spectrum of the dipyrrin 2 (see above). Similar bathochromic shifts of the absorption spectrum upon coordination of a Zn(II) ion were observed in Zn-complexes of bilirubin , a signal of an HN was lacking (which was found at 12.76\u00a0ppm in the spectrum of the dipyrrin 2), consistent with bidentate binding of the dipyrrin ligand 2 to the coordinated Zn(II) ion. Signals of aryl-o hydrogens and aryl-p hydrogen are slightly shifted to lower field while signals of \u03b2-methylene groups and pyrrole-\u03b1 hydrogens are slightly moved to higher field.In the 3 grew from a solution of 3 in CH2Cl2 when n-C6H14 was mixed in slowly at 4\u00a0\u00b0C. The Zn-complex 3 crystalized in the triclinic system with space group P-1 (no. 2). A unit-cell contained four molecules of 3. The crystal structure of 3 showed two bidentate dipyrrin moieties wrapped around one Zn(II) center leading to coordination in a distorted tetrahedral fashion with N\u2013Zn\u2013N angles of about 94, 113, and 121\u00b0 Zn-complexes -cycloaddition reactions. As was recently developed with porphyrinoids, such as tetra-sulfolenoporphyrins were reagent grade commercial chemicals from Fluka and were used as received; EtOAc, dichloromethane, methanol (MeOH), and n-C6H14 were from Acros and were distilled before use. Column chromatography (CC): Fluka silica gel 60 (230\u2013400 mesh). Thin layer chromatography (TLC): Merck 0.25\u00a0mm silica gel 60 plates. Equipment: UV/Vis: Agilent Cary 60 UV\u2013Visible, \u03bbmax in nm (log \u03b5). Fluorescence (FL): Varian Cary Eclipse, \u03bb in nm (rel. intensity); Nuclear magnetic resonance (1H) spectra: Bruker 300 at 298K, chemical shifts (\u03b4) in ppm, with \u03b4 (CHCl3)\u00a0=\u00a07.26\u00a0ppm, signal assignment follows the X-ray numbering scheme. FAB-MS: Finnigan MAT-95, positive ion mode, NOBA matrix; X-ray analyses: data collection on a Nonius Kappa CCD, equipped with graphite mono-chromatized Mo-K\u03b1-radiation (\u03bb\u00a0=\u00a00.71073\u00a0\u00c5) at 233K. Melting point: B\u00fcchi 535.Dichlorodicyano-1 (3.8\u00a0\u03bcmol) +), 894.4 ([M\u20133SO2]+), 830.4 ([M\u20134SO2]+).Dipyrrin 54H62N4O8S4Zn\u00a0\u00d7\u00a0CH2Cl2\u00a0\u00d7\u00a00.75 C6H14, formula weight: 1238.24; temperature 233(2)\u00a0K; radiation wavelength 0.71073\u00a0\u00c5; crystal system triclinic; space group P-1 (no. 2); unit cell dimensions a\u00a0=\u00a018.5998(4)\u00a0\u00c5, b\u00a0=\u00a018.9173(3)\u00a0\u00c5, c\u00a0=\u00a021.6741(5)\u00a0\u00c5, \u03b1\u00a0=\u00a098.771(1)\u00b0, \u03b2\u00a0=\u00a0104.394(1)\u00b0, \u03b3\u00a0=\u00a0109.204(1)\u00b0; volume 6743.1(2) \u00c53; Z\u00a0=\u00a04; density 1.220\u00a0g/cm3; absorption coefficient 0.618\u00a0mm\u22121; crystal size 0.45\u00a0\u00d7\u00a00.25\u00a0\u00d7\u00a00.03\u00a0mm3; F(000) 2606; theta range for data collection 1.335\u00b0\u201324.145\u00b0; index ranges \u221221\u00a0\u2264\u00a0h\u00a0\u2264\u00a021, \u221221\u00a0\u2264\u00a0k\u00a0\u2264\u00a021, \u221224\u00a0\u2264\u00a0l\u00a0\u2264\u00a024; reflections collected 36397; independent reflections 21231 [R(int)\u00a0=\u00a00.0379]; completeness to theta (24.145\u00b0) 98.5\u00a0%; absorption correction none; refinement method Full-matrix least-squares on F2; data/restraints/parameters 21231/29/1489; goodness-of-fit on F2 1.032; final R indices [I\u00a0>2\u03c3(I)] R1\u00a0=\u00a00.0769, wR2\u00a0=\u00a00.1998; R indices R1\u00a0=\u00a00.1148, wR2\u00a0=\u00a00.2209. Crystallographic data of 3 (Zn-(2)2) (excluding structure factors) have been deposited with the Cambridge Crystallographic Data Centre as supplementary publication number CCDC no. 1453954. Copies of the data can be obtained free of charge from the Cambridge Crystallographic Data Centre via http://summary.ccdc.cam.ac.uk/structure-summary-form.Crystallographic data: C"} +{"text": "S(6) ring and therefore establishes a syn relationship for the N atoms. In the crystal, mol\u00adecules are linked by N\u2014H\u22efN hydrogen bonds, generating [100] chains containing alternating A and B mol\u00adecules.The two mol\u00adecules of 2-(2-amino\u00adphen\u00adyl)-1,3-benzoxazole in the asymmetric unit feature an intra\u00admolecular N\u2014H\u22efN hydrogen bond, which closes an 13H10N2O, were grown from a di\u00adchloro\u00admethane/ketone/methanol solvent mixture. It crystallizes with two mol\u00adecules, A and B, in the asymmetric unit with very similar almost planar conformations . Each mol\u00adecule features an intra\u00admolecular N\u2014H\u22efN hydrogen bond, which closes an S(6) ring and therefore establishes a syn relationship for the N atoms. In the crystal, mol\u00adecules are linked by N\u2014H\u22efN hydrogen bonds, generating [100] chains containing alternating A and B mol\u00adecules. Weak aromatic \u03c0\u2013\u03c0 stacking [minimum centroid\u2013centroid separation = 3.6212\u2005(9)\u2005\u00c5] links the chains into a three-dimensional network.Crystals of the title compound, C The oriA and B, respectively. The dihedral angles between the benzene ring and the fused ring system are 0.74\u2005(8) and 0.67\u2005(6)\u00b0 for mol\u00adecules A and B, respectively. The two independent mol\u00adecules are very similar, with an r.m.s. overlay fit of 0.019\u2005\u00c5.The skeleton of each mol\u00adecule is practically planar: to analyse the planarity of the mol\u00adecule we use the torsion angle N3\u2014C2\u2014C8\u2014C9, indicating the rotation of the aromatic ring C8\u2014C13: these angles are \u22121.2\u2005(2) and 0.9\u2005(2)\u00b0 for mol\u00adecules 2 group forms an intra\u00admolecular hydrogen bond of the type N2\u2014H2B\u22efN3 (Table\u00a01A and 2.146\u2005(18)\u2005\u00c5 in mol\u00adecule B, and an inter\u00admolecular N2\u2014H2A\u22efN2 hydrogen bond with a distance of 2.289\u2005(15)\u2005\u00c5 for N2\u2014H2A\u22efN2\u2032 and 2.522\u2005(16)\u2005\u00c5 for N2\u2032\u2014H2A\u2032\u22efN2, forming zigzag chains propagating in the [100] direction and containing alternating A and B mol\u00adecules of o-amino\u00adphenol were added followed by the addition of 0.30\u2005ml (10% mol) of a solution of ZnCl2 (1 M). The mixture was then stirred and heated slowly to reflux temperature during 18\u2005h. The crude reaction product was concentrated on a rotary evaporator with an azeotropic mixture of AcOEt/xylene to obtain a reddish brown solid which was dissolved in EtOAc and washed with 10% aq. NaCl solution. The crude reaction product was purified by column chromatography to give 356\u2005mg (55%) of the amine (I)\u03b3max cm\u22121: 3408 NH2, 3051 C\u2014H(arom), 1624 C=N; : \u03b4 = 6.20 , 6.79 , 7.29 , 7.33 , 757 , 7.72 , 8.09 ; 13C NMR \u03b4 = 108.7, 110.4, 116.3, 116.8, 119.4, 124.3, 124.8, 128.8, 132.5, 141.9, 147.9, 149.3, 163.2 . Analysis calculated for C13H10N2O: C, 74.27; H, 4.79%; Found: C, 74.43; H, 5.05%.500\u2005mg (3.00\u2005mmol) of isatoic anhydride were dissolved in 50\u2005mL of sage 2003; 1H NMR The single crystal used in the experiment was obtained by the method of liquid\u2013liquid diffusion by slow evaporation. The pure compound was dissolved in the minimum amount of di\u00adchloro\u00admethane to be added by the walls of the tube the same amount of acetone followed by methanol. The tube was sealed to leave the solution in a vibration-free environment at room temperature. After a few days, the solution had evaporated, leaving colourless blocks of the title compound.isoU(H) = 1.2eqU(C). Hydrogen atoms of the amine group were found in a difference map and refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015000481/hb7320sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015000481/hb7320Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015000481/hb7320Isup3.cmlSupporting information file. DOI: 1042858CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure of the title compound, the sulfonamide N\u2014H group forms an inter\u00admolecular hydrogen bond to the amine N atom. 15H24N2O2S, was synthesized via a substitution reaction between the enanti\u00adopure -(+)-1,2-di\u00adamino\u00adcyclo\u00adhexane and 2,4,6-tri\u00admethyl\u00adbenzene-1-sulfonyl chloride. The cyclo\u00adhexyl and phenyl substituents are oriented gauche around the sulfonamide S\u2014N bond. In the crystal, mol\u00adecules are linked via N\u2014H\u22efN hydrogen bonds, forming chains propagating along [100].The title compound, C A weak intra\u00admolecular inter\u00adaction is present between the amine H2A atom and the sp2-hybridized sulfonamide N1 atom inter\u00adactions with an H2B\u22efO1 distance of 2.72\u2005\u00c5 between the donor amine N2\u2014H2B and the acceptor sulfonamide O1 atoms can also be noticed within this chain.Mol\u00adecules of the title compound are held together in the solid state by inter\u00admolecular hydrogen-bonding inter\u00adactions between the donor sulfonamide N1\u2014H1 and the acceptor amine N2 atoms Table\u00a01. These hD\u22efH = 0.860\u2005(7)\u2005\u00c5; H\u22efA = 2.160\u2005(8)\u2005\u00c5; D\u22efA = 3.011\u2005(8)\u2005\u00c5; D\u2014H\u22efA = 169.9\u2005(5)\u00b0].As for the racemic crystal FAVHEP, in the model deposited in the CSD there is one inter\u00admolecular hydrogen bond present between a donor sulfonamide N1\u2014H1 and a nearby amine acceptor N atom [et al., 2012et al., 2014et al., 2010trans-1,2-di\u00adamino\u00adcyclo\u00adhexane ring are bonded to a mesitylsulfonamide group. Structure FAVHEP -(+)-1,2-di\u00adamino\u00adcyclo\u00adhexane in 5\u2005ml of CH2Cl2 at 273\u2005K was added a solution of 2,4,6-tri\u00admethyl\u00adbenzene-1-sulfonyl chloride in 5\u2005ml CH2Cl2. After the addition was complete (20\u2005min), the mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was washed with H2O (3 \u00d7 25\u2005ml) and the aqueous layer was back-extracted with CH2Cl2 (20\u2005ml). The combined organic extracts were dried over Na2SO4 and the solvent was removed under reduced pressure. The residue was purified by column chromatography over silica gel (CH2Cl2/EtOAc 1:1 v/v) to afford a pale-yellow\u2013white solid . Part of the purified product was redissolved in CH2Cl2 and after slow evaporation for several days, white large chunky crystals (stained yellow) were formed that were suitable for analysis by X-ray diffraction (m.p. 406\u2013407\u2005K).To a stirred solution of (1Uiso(H) = 1.2Ueq(C) for methine, methyl\u00adene and aryl groups, and Uiso(H) = 1.5Ueq(C) for methyl groups. H atoms bonded directly to N atoms were located in difference-Fourier maps and refined isotropically.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901502191X/gk2646sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901502191X/gk2646Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901502191X/gk2646Isup3.cmlSupporting information file. DOI: 1437453CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "L2(H2O)4]\u00b72H2O , the CdII ion is situated on an inversion centre being coordinated by four aqua mol\u00adecules in the equatorial plane and two deprotonated phospho\u00adnate O atoms from two L ligands in the axial positions in a distorted octa\u00adhedral geometry. Each ligand L exists in a zwitterionic form, and with an intra\u00admolecular O\u2014H\u22efO inter\u00adaction forming an S(6) ring motif and two intra\u00admolecular N\u2014H\u22efO inter\u00adactions each generating an S(5) ring motif. In the crystal, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds link complex mol\u00adecules into a three-dimensional network with voids of 38\u2005\u00c53 filled with ordered lattice water mol\u00adecules, which are also involved in O\u2014H\u22efO hydrogen bonding.In the compound [Cd L2(H2O)4]\u00b72H2O , the CdII ion is situated on an inversion centre being coordinated by four aqua mol\u00adecules in the equatorial plane and two phosphonate O atoms from two deprotonated L ligands in the axial positions in a distorted octa\u00adhedral geometry. The asymmetric unit contains one-half of the complex mol\u00adecule and one lattice water mol\u00adecule. The ligand L exists in a zwitterionic form, with a positive charge on the NH3 group and a negative charge on the O atom of the non-coordinating phospho\u00adnate group, and with an intra\u00admolecular O\u2014H\u22efO inter\u00adaction forming an S(6) ring motif and two intra\u00admolecular N\u2014H\u22efO inter\u00adactions each generating an S(5) ring motif. In the crystal, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds link the complex mol\u00adecules into a three-dimensional network in which the voids of 38\u2005\u00c53 are filled with ordered lattice water mol\u00adecules, which are also involved in O\u2014H\u22efO hydrogen bonding.In the title compound, [Cd All bond lengths and angles in (I)et al., 1991et al., 1997et al., 2005et al., 2006et al., 2007et al., 2007et al., 2009et al., 2010et al., 2010II atom occupies a special position on an inversion centre and shows a slightly distorted octa\u00adhedral coordination environment formed by two phospho\u00adnic O atoms in trans positions and four aqua O atoms in the equatorial plane. The distorted octa\u00adhedral coordination polyhedron is slightly compressed in the axial direction; the Cd1\u2014O2 bond length is 0.1\u2005\u00c5 shorter than the Cd1\u2014O1W and Cd1\u2014O2W bonds. The values of the axial O\u2014Cd\u2014O angles are in the range 80.1\u2005(4)\u201399.9\u2005(4)\u00b0, indicating a significant deviation from ideal values. The ligand L exists in a zwitterionic form, with a positive charge on the NH3 group and a negative charge on the O atom of the non-coordinating phosphonate group, and with an intra\u00admolecular O\u2014H\u22efO inter\u00adaction forming an S(6) ring motif and two intra\u00admolecular N\u2014H\u22efO inter\u00adactions each generating an S(5) ring motif f Table\u00a01.b axis. Inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds at 273\u2005K to produce the title compound (I)1H and 13C{1H} NMR spectra were recorded on a Bruker Bio spin 400 spectrometer.All reactions and manipulations were carried out in air with reagent grade solvents. 1-Amino\u00adethane-1,1-diyldi\u00adphospho\u00adnic acid was prepared according to the literature method of Rukiah & Assaad 2013. The titSpectroscopic data for (I)1H NMR : \u03b4 1.67 . 13C{1H} NMR : \u03b4 20.5 , 54.7 . 31P{1H} NMR : \u03b4 13.61. IR : 3446.2 (NH3), 2351.5 (POH), 1605.0 (O=P\u2014O\u2014H).\u03b11 radiation (\u03bb= 1.54060\u2005\u00c5) selected with an incident-beam curved-crystal Ge(111) monochromator with a linear position-sensitive detector (PSD). The pattern was scanned over the angular range 6.0\u201390.0\u00b0 (2\u03b8). For pattern indexing, extraction of the peak positions was carried out with the program WinPLOTR = 37.1, F(20) = 78.5]. The best estimated monoclinic space group was P21/c.Crystal data, data collection and structure refinement details are summarized in Table\u00a02et al., 1988FULLPROF . These \u03bc\u00b7d values were determined experimentally. The preferred orientation was modelled with 12 coefficients using a spherical harmonics correction = 0.011009 and \u03c72 = 8.940) indicate that the preferred orientation improvement of the refinement is considerable.The powder pattern was subsequently refined with cell and resolution constraints on bond lengths were applied to normal values for these bonds. The final refinement cycles were performed varying isotropic displacement parameters for Cd and water O atoms, and fixed isotropic displacement parameters for P, C, N,O and H atoms. The final Rietveld plot is shown in Fig.\u00a03Before the final refinement, the H atoms of the CH10.1107/S2056989015004028/cv5484sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015004028/cv5484Isup2.rtvRietveld powder data: contains datablock(s) I. DOI: 1051338CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Nature Communications7: Article number: 10269 10.1038/ncomms10269 (2016); Published: 01052015; Updated: 05112016x axis in Supplementary Figure 3 (lower panel). The x axis in this panel should have been labelled \u201822\u2009\u00b0C Mock; 22\u2009\u00b0C GDA; 29\u2009\u00b0C Mock; 29\u2009\u00b0C GDA'. The correct version of this figure appears below.This Article contains errors in the labelling of the Supplementary Figure 3"} +{"text": "The cyclo\u00adhexene ring adopts an envelope conformation in both mol\u00adecules, with the C atom to which is attached the central benzene ring as the flap. The crystal packing, is stabilized by C\u2014H\u22ef\u03c0 inter\u00adactions. 27H26O3, crystallized with two independent mol\u00adecules (A and B) in the asymmetric unit. In mol\u00adecule A, the plane of the central benzene ring forms dihedral angles of 75.78\u2005(14) and 52.75\u2005(16)\u00b0 with that of the terminal benzene rings, and the dihedral angle between the planes of the terminal benzene rings is 51.49\u2005(17)\u00b0. The corresponding values for mol\u00adecule B are 75.18\u2005(14), 58.11\u2005(16) and 47.91\u2005(16)\u00b0, respectively. The cyclo\u00adhexene ring adopts an envelope conformation in both mol\u00adecules, with the C atom to which is attached the central benzene ring as the flap. The crystal packing, is stabilized by C\u2014H\u22ef\u03c0 inter\u00adactions.The title compound, C Cyclo\u00adhexenone and cyclo\u00adhexenone derivatives are known for anti-inflammatory and analgesic activities in the asymmetric unit and 1.224\u2005(3)\u2005\u00c5, respectively, indicate double-bond character. In both mol\u00adecules, the C\u2014O bond lengths are in the range 1.362\u2005(3)\u20131.414\u2005(4)\u2005\u00c5 and represent single-bond character. In mol\u00adecule A, the torsion angles C5\u2014C4\u2014C7\u2014C8 = 69.2\u2005(4)\u00b0 and C24\u2014C9\u2014C10\u2014C11 = \u221216.0\u2005(4)\u00b0 show that the benzyl\u00adoxyphenyl and meth\u00adoxy\u00adphenyl groups have a +sc and -sp orientation with respect to the cyclo\u00adhexene moiety. The arrangement in mol\u00adecule B is slightly different, with torsion angles C5A\u2014C4A\u2014C7A\u2014C8A = 111.5\u2005(3)\u00b0 and C24A\u2014C9A\u2014C10A\u2014C11A = 20.8\u2005(4)\u00b0. The cyclo\u00adhexene ring in both mol\u00adecules adopts an envelope conformation with atoms C7 and C7A as the flaps in mol\u00adecules A and B, respectively.The title compound crystallized with two mol\u00adecules -3-(4-Benzyl\u00adoxyphen\u00adyl)-1-(4-meth\u00adoxy\u00adphen\u00adyl)prop-2-en-1-one was synthesized following the literature procedure of Ezhilarasi et al. -3-(4-benzyl\u00adoxyphen\u00adyl)-1-(4-meth\u00adoxy\u00adphen\u00adyl)prop-2-en-1-one (0.01\u2005mol) and ethyl methyl ketone (0.01\u2005mol) were refluxed in absolute alcohol (50\u2005ml) in the presence of 10% sodium hydroxide solution (10\u2005ml) for 1\u2005h in an oil bath. The reaction mixture was then cooled and the precipitate obtained filtered, washed with distilled water and dried. The crude product was recrystallized twice from absolute alcohol (yield 80%), giving yellow block-like crystals. = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989014025390/su5016sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989014025390/su5016Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989014025390/su5016Isup3.cmlSupporting information file. DOI: 1035112CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The sulfonate group is disordered over two positions with occupancy factors of 0.655\u2005(5) and 0.345\u2005(5). The hexa\u00adaqua\u00adnickel(II) cation inter\u00adacts through hydrogen bonding with eight QOH mol\u00adecules and two water mol\u00adecules. The six-membered rings of quinoline show \u03c0\u2013\u03c0 stacking [centroid-to-centroid distances of 3.679\u2005(2)\u2005\u00c5 and 3.714\u2005(2)\u2005\u00c5].The asymmetric unit of the title compound, [Ni(H Quinine, cinchonine, chloro\u00adquine, plasmoquine and acriquine, for instance, are known to be able to cure malaria has been synthesized from eugenol and its anti\u00adbacterial activities have been reported . As quinoline rings are known to complex with metal ions, the formation of a complex between QOH and NiII was studied. The reaction product, however, could not be characterized unambiguously by IR or 1H NMR spectroscopic methods. The spectroscopic data are different from those obtained for free QOH and in favour of a deprotonated carb\u00adoxy\u00adlic acid group, but give no indication about a possible complex formation. X-ray diffraction now shows that QOH is not complexing directly with NiII.Recently, the new quinoline derivative 6-hy\u00addroxy-3-sulfoquinolin-7-yloxyacetic 6]2+ is located about an inversion center and has an octa\u00adhedral volume of 11.629\u2005\u00c53 with Ni\u2014O bond lengths between 2.034\u2005(3) and 2.106\u2005(2)\u2005\u00c5.The structure determination shows that the carboxyl group of rm Fig.\u00a01, which wQOH moieties and two water mol\u00adecules through O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonding (Table\u00a01The hexa\u00adaqua\u00adnickel(II) cation plays the role of glue in the crystal packing. In total, it inter\u00adacts with eight g Table\u00a01.Cg1\u22efCg1ix = 3.679\u2005(2)\u2005\u00c5, Cg1\u22efCg2ix = 3.714\u2005(2)\u2005\u00c5; Cg1 and Cg2 are the centroids of the rings C12/C13/N14/C15\u2013C17 and C15/C16/C18\u2013C21, respectively; symmetry code: (ix) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01; Fig.\u00a03Furthermore, \u03c0\u2013\u03c0 stacking between the quinoline rings results in the formation of inversion dimers [QOH mol\u00adecule and furthermore with the sulfonate group (O7) of a second QOH mol\u00adecule and the hexa\u00adaqua complex (O2). Whereas hydroxyl group O23\u2014H23 only inter\u00adacts with water mol\u00adecule O29, the second hydroxyl group O22\u2014H22 is involved in the formation of another type of inversion dimers through C\u2014H\u22efO hydrogen bonding and inter\u00adacts with a sulfonate group (O8) (Table\u00a01Lattice water mol\u00adecule O29 inter\u00adacts with the carboxyl\u00adate (O27) and hydroxyl (O23) groups of a neighboring ) Table\u00a01.A search of the Cambridge Structural Database was synthesized and further transformed to 5,6-dihy\u00addroxy-3-sulfoquinolin-7-yloxyacetic acid (QOH) according to a procedure described by Dinh et al. in ethanol\u2013water was added dropwise to a solution of QOH in ethanol\u2013water . The obtained solution was stirred for three hours, at 313\u2013323\u2005K, during reflux. A few days later, the green\u2013yellow precipitate was collected by filtration, washed consecutively with ethanol and diethyl ether and dried in vacuo. The obtained crystals are soluble in water and DMSO, but only slightly soluble in ethanol, acetone and chloro\u00adform. The yield was 65%. Single crystals suitable for X-ray investigation were obtained by slow evaporation from a ethanol\u2013water (1:1 v/v) solution at room temperature. IR : 3420 (\u03bdOH); 3080, 2918 (\u03bdC-H); 1620 (\u03bdCOOas); 1426(\u03bdCOOs); 1528 (\u03bdC=Cring or \u03bdC=N); 466 (\u03bdNi-O). 1H NMR : \u03b4 8.74 , 8.17 , 7.2 , 4.64 ; : \u03b4 9.26 , 9.01 , 7.01 , 4.80 .A solution containing NiClB, H3B, H4B, H14, H29A and H29B were located in difference Fourier maps. All other H atoms were placed at idealized positions and refined in riding mode, with C\u2014H distances of 0.95 (aromatic) and 0.99\u2005\u00c5 (methyl\u00adene), and O\u2014H distances of 0.84\u2005\u00c5. The H atoms of water mol\u00adecule O29 were refined with an O\u2014H distance restraint of 0.85\u2005\u00c5 and H\u22efH distance restraint of 1.39\u2005\u00c5. For all H atoms, Uiso(H) values were assigned as 1.2Ueq of the parent atoms (1.5Ueq for H22 and H23). The SO3 group is disordered over two positions, the occupancy ratio refines to 0.655\u2005(5):0.345\u2005(5) for part 1 and part 2 , respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015015662/vn2096sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015015662/vn2096Isup2.hklStructure factors: contains datablock(s) I. DOI: 1419884CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports5 Article number: 14289; 10.1038/srep14289 published online: 09232015; updated: 12182015.In Supplementary Figure 1a, the Linzizong volcanic rock samples \u201812LZ14-1\u2019 and \u201812LZ13-1\u2019 should read \u201813LZ14-1\u2019 and \u201813LZ13-1\u2019 respectively. The correct Supplementary Figure 1a appears below as In Table S1, samples \u201812LZ13-1@02\u2019 and \u201812LZ14-1@02\u2019 should read \u201813LZ13-1@02\u2019 and \u201813LZ14-1@02\u2019 respectively."} +{"text": "The angles at the NiII cation, which lies on a twofold rotation axis, are Cl\u2014Ni\u2014Cl = 115.58\u2005(3)\u00b0 and S\u2014Ni\u2014S = 94.55\u2005(3)\u00b0. All other angles at the central NiII atom range from 109.46\u2005(2) to 112.96\u2005(2)\u00b0. The C\u2014S\u2014Ni angle is 99.91\u2005(6)\u00b0. The planes of two imidazolium rings make a dihedral angle of 70.56\u2005(6)\u00b0.In the mol\u00adecular structure of the title compound, [NiCl DOI: 10.1107/S2056989015012281/hp2071Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015012281/hp2071fig1.tif. DOI: Mol\u00adecular structure of the title compound with anisotropic displacement parameters drawn at the 50% probability level.1408996CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit consists of one Cd2+ cation, two thio\u00adcyanate anions and two 4-(hy\u00addroxy\u00admeth\u00adyl)pyridine ligands in general positions. Two Cd2+ cations are linked by two \u03bc-1,3 N- and S-bonding thio\u00adycanate anions into dimers which are further linked into branched chains along [100] by two \u03bc-1,6 N- and O-bonding 4-(hy\u00addroxy\u00admeth\u00adyl)pyridine ligands. One additional N-bonded 4-(hy\u00addroxy\u00admeth\u00adyl)pyridine ligand and one additional N-bonded thio\u00adcyanate anion are only terminally bonded to the metal cation. Inter\u00adchain O\u2014H\u22efS hydrogen bonds between the hy\u00addroxy H atoms and one of the thio\u00adcyanate S atoms connect the chains into a three-dimensional network.The crystal structure of the title compound, [Cd(NCS) DOI: 10.1107/S2056989015008890/wm5156Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015008890/wm5156fig1.tifx y z x y z . DOI: x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01; (ii) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01.]Part of the crystal structure of the title compound with labelling and displacement ellipsoids drawn at the 50% probability level. [Symmetry codes: (i) \u2212Click here for additional data file.10.1107/S2056989015008890/wm5156fig2.tif. DOI: Crystal structure of the title compound in a view approximately along [001]. Inter\u00admolecular O\u2014H\u22efS hydrogen bonding is shown as dashed lines; the disordered pyridine rings are omitted for clarity.1063786CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Solvent-dependent outcomes are noted in co-crystallization experiments between DABCO and 4-nitrobenzoic acid with mono- and diprotonated forms of DABCO are isolated. 6H13N2+\u00b7C7H4NO4\u2212\u00b72H2O, (1), whereas from methanol, the salt C6H14N22+\u00b72C7H4NO4\u2212, (2), was isolated. In (1), the cation and anion are linked by a strong N\u2014H\u22efO hydrogen bond, and the carboxyl\u00adate anion is close to planar [dihedral angle between terminal residues = 6.83\u2005(9)\u00b0]. In (2), a three-ion aggregate is assembled by two N\u2014H\u22efO hydrogen bonds, and the carboxyl\u00adate anions are again close to planar [dihedral angles between terminal residues = 1.7\u2005(3) and 5.9\u2005(3)\u00b0]. Through the inter\u00advention of solvent water mol\u00adecules, which self-assemble into helical supra\u00admolecular chains along the b axis, the three-dimensional architecture in (1) is stabilized by water\u2013DABCO O\u2014H\u22efN and water\u2013carboxyl\u00adate O\u2014H\u22efO hydrogen bonds, with additional stability afforded by C\u2014H\u22efO inter\u00adactions. The global crystal structure comprises alternating layers of water mol\u00adecules and ion pairs stacked along the c axis. In the crystal of (2), the three-ion aggregates are assembled into a three-dimensional architecture by a large number of methyl\u00adene\u2013carboxyl\u00adate/nitro C\u2014H\u22efO inter\u00adactions as well as \u03c0\u2013\u03c0 contacts between inversion-related benzene rings [inter-centroid distances = 3.5644\u2005(16) and 3.6527\u2005(16)\u2005\u00c5]. The cations and anions assemble into alternating layers along the c axis.The 1:1 co-crystallization of 1,4-di\u00adaza\u00adbicyclo\u00ad[2.2.2]octane (DABCO) with 4-nitro\u00adbenzoic acid in ethanol\u2013water (3/1) gave the salt dihydrate C In this context, it is not surprising that a search of the Cambridge Structural Database comprises a 1,4-di\u00adaza\u00adbicyclo\u00ad[2.2.2]octan-1-ium mono-cation, a 4-nitro\u00adbenzoate anion and two water mol\u00adecules of hydration Fig.\u00a01. The mosi.e. 1.281\u2005(3) and 1.228\u2005(3)\u2005\u00c5, is slightly greater than that in C8\u2014O5, O6 of 1.273\u2005(3) and 1.231\u2005(3)\u2005\u00c5, respectively. In each case the longer bond forms a strong N\u2014H\u22efO hydrogen bond and 8.7\u2005(2)\u00b0, respectively, and the dihedral angle between the terminal groups is 1.7\u2005(3)\u00b0. The comparable angles for the O5-containing anion are 2.2\u2005(3), 7.4\u2005(2) and 5.9\u2005(3)\u00b0, respectively. As discussed below in Supra\u00admolecular features, the ions participate in strong N\u2014H\u22efO hydrogen bonds, forming a three-ion aggregate comprises a 1,4-di\u00adaza\u00adbicyclo\u00ad[2.2.2]octane-1,4-diium di-cation and two 4-nitro\u00adbenzoate anions Fig.\u00a02. In the d Table\u00a02. In ordete Fig.\u00a02 in whichb axis. Adjacent columns stack along the a axis to form layers in the ab plane. The layers are inter\u00adspersed by layers of water mol\u00adecules which self-assemble into helical chains along the b axis, where each independent water mol\u00adecule donates and accepts a water-O\u2014H\u22efO(water) hydrogen bond (Table\u00a01In (1), the cation and anion are linked by a strong N2\u2014H\u22efO1 hydrogen bond Fig.\u00a01. The twod Table\u00a01. This led Table\u00a01.W\u2014H\u22efN3 and water-OW2\u2014H\u22efO2(carboxyl\u00adate). Additional stability to the supra\u00admolecular assembly is afforded by methyl\u00adene-C\u2014H\u22efO2(carboxyl\u00adate) and O2W(water) inter\u00adactions; it is noteworthy that both of the former inter\u00adactions involve hydrogen atoms derived from the same methyl\u00adene-C12 atom \u2005\u00c5 for inter\u00adactions between the C2\u2013C8 rings and 3.6527\u2005(16)\u2005\u00c5 between C9\u2013C14 rings . An alternate description of the global crystal packing is based on alternating of layers of cations and layers of anions along the c axis , the di-cation is linked to two anions ds Fig.\u00a04. Globall\u00c5 Table\u00a02. All int\u00c5 Table\u00a02. Additiois Fig.\u00a05.Chemical context, there are 57 species in the crystallographic literature containing DABCO or its mono- or diprotonated forms and a carb\u00adoxy\u00adlic acid or carboxyl\u00adate anion. In fact, co-crystals are rare, being around 10% of all structures. Co-crystals are formed with several di\u00adcarb\u00adoxy\u00adlic acids where the functional groups are separated by long chains of over four carbon atoms , was mixed with 4-nitro\u00adbenzoic acid in a solution containing ethanol (30\u2005ml) and water (10\u2005ml). The solution was heated for 2\u2005h at 350\u2005K. The mixture was then left for slow evaporation and colourless crystals of (1) formed after 4 days. In a similar experiment, 1,4-di\u00adaza\u00adbicyclo\u00ad[2.2.2]octane was mixed with 4-nitro\u00adbenzoic acid in a solution of methanol (50\u2005ml). The solution was heated for 2\u2005h at 345\u2005K. The mixture was then left for slow evaporation and colourless crystals of (2) formed after 4 days.Uiso(H) = 1.2Ueq(C). The N-bound H-atoms were located in a difference Fourier map but were refined with a distance restraint: N\u2014H = 0.88\u2005(1)\u2005\u00c5 with Uiso(H) = 1.2Ueq(N). For (1), the water-bound H atoms were refined with distance restraints: O\u2014H = 0.84\u2005(1) and H\u22efH = 1.39\u2005(2)\u2005\u00c5 with Uiso(H) =1.5Ueq(O). For (2), the maximum and minimum residual electron density peaks of 0.60 and 0.58\u2005e\u2005\u00c5\u22123, respectively, were located 0.81 and 0.10\u2005\u00c5 from atoms O5 and H4N, respectively. In order to confirm the location of the N-bound H atoms, in a separate refinement these were refined without distance restraints. For (1), the N2\u2014H2N bond length was 0.948\u2005(17)\u2005\u00c5. For (2), the N3\u2014H3N and N4\u2014H4N bond lengths were 0.93\u2005(4) and 1.08\u2005(3)\u2005\u00c5, respectively. In the refinement of (1), one reflection, i.e. (180), was omitted from the refinement owing to poor agreement. For the same reasons, the following reflections were omitted from the final refinement of (2): (550), 1, 2, global. DOI: 10.1107/S1600536814011532/su00051sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S1600536814011532/su00052sup3.hklStructure factors: contains datablock(s) 2. DOI: Click here for additional data file.10.1107/S1600536814011532/su00051sup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S1600536814011532/su00052sup5.cmlSupporting information file. DOI: 1004283, 1004284CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III ion has a distorted O5Cl octa\u00adhedral coordination environment defined by two bidentate 2-eth\u00adoxy-6-formyl\u00adphenolato ligands, one Cl atom and one water mol\u00adecule. In the crystal, O\u2014H\u22efO hydrogen bonds link the two independent mol\u00adecules to form a dimer while the solvent mol\u00adecule is linked to the complex mol\u00adecule by a weak C\u2014H\u22efO hydrogen bond. Further weak C\u2014H\u22efO inter\u00adactions along with weak C\u2014H\u22efCl hydrogen bonds link the components into chains parallel to [001].In both complex mol\u00adecules in the asymmetric unit, the Fe L)2Cl(H2O)]\u00b70.5CH3CN, , there are two independent complex mol\u00adecules and one aceto\u00adnitrile solvent mol\u00adecule in the asymmetric unit. In each complex mol\u00adecule, the FeIII ion has a distorted O5Cl octa\u00adhedral coordination environment defined by two bidentate 2-eth\u00adoxy-6-formyl\u00adphenolato ligands, one Cl atom and one water mol\u00adecule. In the crystal, O\u2014H\u22efO hydrogen bonds link the two independent mol\u00adecules to form a dimer. The solvent mol\u00adecule is linked to the complex mol\u00adecule by a weak C\u2014H\u22efO hydrogen bond. Further weak C\u2014H\u22efO inter\u00adactions along with weak C\u2014H\u22efCl hydrogen bonds link the components into chains parallel to [001].In the title compound, [Fe( Those complexes display dominant ferromagnetic inter\u00adactions between metal ions.Recently, we have investigated the coordination behavior of the tridentate 2-hy\u00addroxy-benzaldehyde ramification ligand 3-eth\u00adoxy-2-hy\u00addroxy-benzaldehyde and reported two heterometallic polymers [ZnNa(ehbd)L)2Cl(H2O)]\u00b70.5CH3CN (HL = C9H10O3), was prepared similarly to the cubane cluster [Ni4(\u03bc3-OMe)4(heb)4(MeOH)1.05(H2O)2.95] \u00b76H2O was replaced by FeCl3\u00b76H2O in an attempt to prepare a cubane-type iron cluster. The crystals obtained, however, were those of the title mononuclear FeIII complex.The title compound, [Fe(L)2Cl(H2O)] mol\u00adecules and a aceto\u00adnitrile solvent mol\u00adecule. One of the independent mol\u00adecules is shown in Fig.\u00a01III ion is coordinated by four O atoms from two different \u2212L ligands, one Cl\u2212 ion and one terminal water mol\u00adecule, forming a distorted octa\u00adhedral geometry. The Fe\u2014O bond lengths are in the range 1.909\u2005(2)\u20132.157\u2005(2)\u2005\u00c5 (Table\u00a01trans-angles at the FeIII atom lie in the range 169.4\u2005(1)\u2013171.4\u2005(1)\u00b0, the cis-angles vary from 81.6\u2005(1) to 99.9\u2005(1)\u00b0. The \u2212L ligand displays a \u03bc1:\u03ba1:\u03ba1 coordination mode, which is the same as that of [Ni4(\u03bc3-OMe)4(heb)4(MeOH)1.05(H2O)2.95] 2(N3)6]n \u00c5 is observed, where Cg1 and Cg2 are the centroids defined by ring atoms C1\u2013C6 and C19\u2013C24, respectively. The solvent mol\u00adecule is linked to the complex mol\u00adecule by a weak C\u2014H\u22efO hydrogen bond. Further weak C\u2014H\u22efO inter\u00adaction along with weak C\u2014H\u22efCl hydrogen bonds link the components into chains parallel to [001] , 3-eth\u00adoxy-2-hy\u00addroxy-benzaldehyde , methanol (5\u2005mL) and aceto\u00adnitrile (5\u2005mL), with a pH adjusted to 7.5 by addition of tri\u00adethyl\u00adamine, was poured into a Teflon-lined autoclave (15\u2005mL) and then heated at 413K for 3 days. Black crystals of the title compound were collected by filtration, washed with methanol and dried in air. Phase pure crystals were obtained by manual separation .A mixture of FeCl2) or 0.97\u2005\u00c5 (CH3) with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(Cmeth\u00adyl). H atoms bonded to O atoms were included with O\u2014H = 0.84\u20130.85\u2005\u00c5 and with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S1600536814021205/lh5728sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814021205/lh5728Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814021205/lh5728Isup3.cdxSupporting information file. DOI: 1025672CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Hypoxia inducible factor 1\u03b1 (HIF-1\u03b1) is a stress-responsive transcription factor to hypoxia and its expression is correlated to tumor progression and angiogenesis. Several single nucleotide polymorphisms (SNPs) of HIF-1\u03b1 gene in the oxygen-dependent degradation (ODD) domain was reportedly associated with increased HIF-1\u03b1 activity.In this study, we focused on the relationship between SNP 1772 C\u2009>\u2009T (rs11549465) of HIF-1\u03b1 gene and its breast cancer risk, as well as its correlation with HIF-1\u03b1 expression and tumor angiogenesis. Ninety six breast cancer patients and 120 age-matched controls were enrolled. We found that 1772\u00a0T allele of HIF-1\u03b1 gene was associated with increased breast cancer risk . This SNP was not associated with clinicopathologic features of angiogenesis such as VEGF activity and the micro-vessel density and survival of breast cancer patients.Taken together, the 1772 C\u2009>\u2009T of HIF-1\u03b1 gene is a potential biomarker for breast cancer susceptibility. Single nucleotide polymorphisms (SNPs), the most common variants in human genome , are popTumor hypoxia is common in tumorigenesis. Hypoxia inducible factor-1 (HIF-1) is a crucial transcription factor in cellular response to tumor hypoxia and is considered as an adverse prognostic factor in breast cancers -14. AddiRecent studies demonstrated that another SNP located in ODD of HIF-1\u03b1, 1772 C\u2009>\u2009T (rs11549465), may lead to an amino acid change from proline 582 to serine (P582S) and are reportedly associated with renal ,19, headThe purpose of this study is to investigate the association between SNP 1772 C\u2009>\u2009T of the HIF-1\u03b1 gene in breast cancer patients and healthy control subjects. Furthermore, HIF-1 has been reported to transactivate many oxygen responsive genes such as vascular endothelial growth factor (VEGF) . TherefoBetween 1991 and 2001, a total of 96 randomly-selected female patients with breast cancer at Kaohsiung Medical University Hospital, Kaohsiung, Taiwan, were enrolled in this study. All patients underwent a standard modified radical mastectomy. Ninety-four patients received adjuvant systemic chemotherapy with 6\u00a0cycles of 5-fluorourcil, doxorubicin and cyclophosphamide. After completion of chemotherapy, all patients received hormone therapy with tamoxifen and 92 patients received radiation therapy. The principle of treatment was followed as described previously . We coll2, 0.2\u00a0\u03bcl of 10\u00a0mM dNTP each, 0.6\u00a0\u03bcl DMSO, 0.14\u00a0\u03bcl of Taq enzyme, 0.12\u00a0\u03bcl of 350\u00a0\u03bcg/ml primers mix (1:1), 2\u00a0\u03bcl DNA extracts and 5.64\u00a0\u03bcl distilled water. PCR was performed with the following protocol: 94\u00b0C (1\u00a0min); 4\u00a0cycles of 94\u00b0C (15\u00a0s), 64\u00b0C (15\u00a0s), 70\u00b0C (8\u00a0s); 4\u00a0cycles of 94\u00b0C (15\u00a0s), 61\u00b0C (15\u00a0s), 70\u00b0C (8\u00a0s); 4\u00a0cycles of 94\u00b0C (15\u00a0s), 58\u00b0C (15\u00a0s), 70\u00b0C (8\u00a0s); 60\u00a0cycles of 94\u00b0C for (15\u00a0s), 55\u00b0C (15\u00a0s), 70\u00b0C (8\u00a0s); 94\u00b0C (1\u00a0min) and 60\u00b0C (5\u00a0min). The available restriction enzyme for HIF-1\u03b1 1772 C\u2009>\u2009T (rs11549465) was retrieved from the SNP-RFLP freeware [Hph I restriction enzyme (NEB) at 37\u00b0C for overnight and then they were subjected to 3% agarose electrophoresis and stained with SYBR Safe\u2122 DNA gel stain (Invitrogen) for visualization of the PCR-restriction fragment length polymorphism (PCR-RFLP) patterns.Genomic DNA was isolated from paraffin-embedded tumor tissues of surgical specimens and peripheral blood of 120 normal controls as described ,28. The freeware -31. PCR Typical patterns of genotyping by PCR-RFLP have confirmed by sequencing. DNA amplicon from PCR reaction was purified using a MiniElute PCR purification kit (Qiagen) for commStreptoavidin-biotin based immunohistochemical staining (IHC) was performed to detect HIF-1\u03b1 and VEGF protein levels as previously described . ImmunorMicrovessel density (MVD) represents tumor angiogenesis by using immunostaining of endothelial cells with monocloncal antibody, recognizing the CD31 endothelial glycoprotein. Each slide was scanned at low magnification (\u00d7 100) to identify the four areas of high density of microvessels (hotspots). The number of stained vessels per in each hotspot was counted at high power fields (\u00d7 400). Any stained endothelia cell was considered as a countable single microvessel. Large vessels with thick muscular walls were excluded. MVD was classified as either low (\u226635.0) or high (>35.0/high power field (HPF)); 35.0 was the median value.p values smaller than 0.05 are regarded as significance.Statistical significance was evaluated by the chi-square test and Fisher exact test. Overall survival curves were analyzed by the Kaplan-Meier method, and differences between the curves were analyzed by log-rank test. The p\u2009=\u20090. 22).In Table\u00a0In Figure\u00a0p\u2009<\u20090.001). The allele frequencies in controls and cancer patients were 232 C (97%)/8\u00a0T (3%) and 127 C (66%)/65\u00a0T (34%), respectively. The T-allele distribution in breast cancer patients differed significantly from that of controls .Based on PCR-RFLP analysis, the genotype distribution of control group was 116 CC (97%), 0 CT (0%) and 4 TT (3%). In contrast, the genotype distributions of breast cancer patients were 53 CC (55%), 21 CT (22%), and 22 TT (23%). The genotype distribution in breast cancer patients differed significantly from that of controls , T-stage (p\u2009=\u20090.303), N-stage (p\u2009=\u20090.936), local recurrence (p\u2009=\u20090.817), distant metastasis (p\u2009=\u20090.572), HIF-1\u03b1 expression (p\u2009=\u20090.311), VEGF expression (p\u2009=\u20090.375) and microvessel density (p\u2009=\u20090.211).When connecting the results of these stainings with HIF-1\u03b1 genotypes with clinicopathological analysis Table\u00a0, there wp\u2009<\u20090.001) and microvessel density (Exp. (B)\u2009=\u20092.6082, p\u2009<\u20090.05) were the most influential factors and overall survival by multi-variable analyses. Similarly, Kaplan-Meier analysis and overall survival curves , respectively.In Table\u00a0rs Table\u00a0. HoweverThe SNP 1772 C\u2009>\u2009T of HIF-1\u03b1 gene chosen in current study are located within ODD of the HIF-1\u03b1. We found that T allele of the SNP 1772 C\u2009>\u2009T (P582S) of HIF-1\u03b1 gene was significantly higher in 96 breast cancer patients than in 120 controls. In contrast, the association results of SNP 1772 C\u2009>\u2009T of HIF-1\u03b1 gene with different kinds of cancers were not consistent in literature review. For example, the SNP 1772 C\u2009>\u2009T of HIF-1\u03b1 gene was detected in several cancers -21,23 buWithin ODD of the HIF-1\u03b1, proline residues 402 and 564 were reported to independently determine tightly binding to the von Hippel-Lindau (VHL) protein for HIF-1\u03b1 ubiquitination and degradation under nonhypoxia condition ,36-39. IFurthermore, the genotypes of SNP 1772 C\u2009>\u2009T of HIF-1\u03b1 gene are not significantly associated with clinicopathologic characteristics and clinical outcome of breast cancer (Table\u00a0p\u2009=\u20090.059) and disease-free survival (p\u2009=\u20090.110) [p\u2009=\u20090.0732) but weak association with overall survival (p\u2009=\u20090.3225) . SimilarThe phenomena mentioned above may be partly explained by the multigene theory for carcinogenesis . FurtherTaken together, SNP 1772 C\u2009>\u2009T (P582S) of HIF-1\u03b1 gene confers significant association with breast cancer risk but it show no association with the clinicopathologic features and survival of breast cancer patients."} +{"text": "In the title salt, the cations and anions are linked through O\u2014H\u22efO,N\u2014H\u22efO, N\u2014H\u22efN and \u03c0\u2013\u03c0 stacking inter\u00adactions, forming double layers parallel to (101). Weak C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds connect the double layers into a three-dimensional network. 6H9ClN3+\u00b7C6H3O4S\u2212, the cations and anions are linked via O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming R66(37) ring motifs that are inter\u00adconnected with each other, producing sheets. Separate parallel inversion-related sheets are linked through N\u2014H\u22efN and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.5414\u2005(13)\u2005\u00c5], forming double layers parallel to (101). Weak C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds, as well as C\u2014H\u22ef\u03c0 inter\u00adactions, connect the double layers into a three-dimensional network.In the title salt, C The mixture was cooled slowly and kept at room temperature. After a few days colourless plate-like crystals were obtained.3). Isotropic displacement parameters for these atoms were set to 1.2 (CH) or 1.5 (CH3) times Ueq of the parent atom. Idealized Me H atoms were refined as rotating groups. There are larger than expected residual density peaks close to the Cl and S atoms but these are not chemically sensible and are assumed to be related to the quality of the crystal.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016010148/lh5814sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016010148/lh5814Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016010148/lh5814Isup3.cmlSupporting information file. DOI: 1486940CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal of (I), pairs of [Fe(CN)4]\u2212 units are linked together through \u03c0\u2013\u03c0 stacking between the pyridyl rings of the 2,2\u2032-bipy ligands to form a graphite-like structure parallel to the ab plane. The three independent water mol\u00adecules are hydrogen-bonded alternately with each other, forming a ladder chain structure with R44(8) and R66(12) graph-set ring motifs, while the disordered [N(CH3)4]+ cations lie above and below the water chains, and the packing is stabilized by weak C\u2014H\u22efO hydrogen bonds. The water chains are further linked with adjacent sheets into a three-dimensional network via O\u2014H\u22efO hydrogen bonds involving the lattice water mol\u00adecules and the N atoms of terminal cyanide groups of the [Fe(CN)4]\u2212 building blocks, forming an R44(12) ring motif. Compound (II) features a two-dimensional {[Fe(CN)4Cd(en)2]}n layer structure (en is ethyl\u00adenedi\u00adamine) extending parallel to (010) and constructed from {[Fe(CN)4Cd(en)]}n chains inter\u00adlinked by bridging en ligands at the Cd atoms. Classical O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds involving the lattice water mol\u00adecule and N atoms of terminal cyanide groups and the N\u2014H groups of the en ligands are observed within the layers. The layers are further connected via \u03c0\u2013\u03c0 stacking inter\u00adactions between adjacent pyridine rings of the 2,2\u2032-bipy ligands, completing a three-dimensional supra\u00admolecular structure.The crystal structures of the building block tetra\u00admethyl\u00adammonium (2,2\u2032-bi\u00adpyridine-\u03ba M(CN)6]n\u2212 (n = 2\u20134), have been used extensively as building blocks for the design and construction of a large number of high-dimensional cyanide-bridged bimetallic coordination polymers because of their ability to act as multidentate ligands to link numerous metal atoms through all six cyanide groups 6]3\u2013 with octa\u00adhedrally coordinated transition metal ions such as 2,2\u2032-bi\u00adpyridine or 1,10-phenanthroline in cyanide-containing building blocks of general formula [M(L)(CN)4]n\u2212 instead of [M(CN)6]n\u2212 has been a recent development in the field of low-dimensionality cyanide-bridged bimetallic coordination compounds 4][Fe(CN)4(C10H8N2)]\u00b73H2O (I)4)(C10H8N2)(C2H8N2)2]\u00b7H2O (II)As part of our search for novel cyanide-bridged bimetallic coordination polymers, we herein describe the synthesis and crystal structure of [N4]+ and three water mol\u00adecules, as displayed in Fig.\u00a01III ion is coordinated by two nitro\u00adgen atoms from one 2,2\u2032-bipy ligand and four cyanide carbon atoms in a distorted octa\u00adhedral geometry. This distortion around the metal atom is defined by the sum of the octa\u00adhedral angular deviations from 90\u00b0 (\u03a3), in which the trigonal distortion angle = 0 for a perfect octa\u00adhedron \u00b0. The three trans angles are bent slightly from the ideal value of 180\u00b0. The iron atom and terminal cyanido groups, viz. [Fe1\u2014C3\u2261N3 = 178.7\u2005(3) and Fe1\u2014C4\u2261N4 = 179.8\u2005(4)\u00b0] are almost linear compared to the iron atom and the corresponding equatorial cyano groups [viz. Fe1\u2014C1\u2014N1 = 175.8\u2005(4) and Fe1\u2014C2\u2014N2 = 176.6\u2005(4)\u00b0]. This difference is probably caused by hydrogen bonding (see below). The Fe\u2014C bond lengths range from 1.917\u2005(4) to 1.969\u2005(4)\u2005\u00c5, whereas the Fe\u2014N bond lengths are 1.981\u2005(3) and 1.985\u2005(3)\u2005\u00c5. The whole mol\u00adecule of 2,2\u2032-bipy ligand is planar with an r.m.s. deviation of 0.016\u2005\u00c5; the dihedral angle between the two pyridyl rings is 1.57\u2005(18)\u00b0. Bond lengths and angles within the [Fe(CN)4]\u2212 anion in (I)4]\u00b7H2O (CN)4]\u00b7H2O (CN)4]\u00b7CH3CN consists of one [Fe(CN)III precursor was reduced to FeII under the crystallization conditions. The asymmetric unit contains half each of an [Fe(CN)4]\u2212 anion and a [Cd(en)2]2+ cation, with the mol\u00adecules lying across twofold rotation axes, Fig.\u00a02II ion is a distorted octa\u00adhedron with a \u03a3 of 28.90\u00b0. The Fe\u2014C\u2014N angles for both bridging [Fe1\u2014C1\u2014N1 = 178.15\u2005(14)\u00b0] and terminal [Fe1\u2014C2\u2014N2 = 176.85\u2005(16)\u00b0] cyanide groups deviate slightly from strict linearity. The Fe\u2014Ccyanide bond lengths at 1.8950\u2005(16) and 1.9363\u2005(17)\u2005\u00c5 are slightly shorter than the Fe\u2014N2,2\u2032-bipy bond length, 1.9976\u2005(14)\u2005\u00c5. The CdII ion is six-coordinated by two N atoms from two cyanide groups, two N atoms from a chelating en ligand and two N atoms from two different bridging en ligands in a highly distorted octa\u00adhedral geometry with a \u03a3 of 108.08\u00b0. The Cd\u2014N bond lengths and the N\u2014Cd\u2014N bond angles in (II)4N)[{Fe(CN)6}3{Cd(en)}4] 2] }3]\u00b74H2O ]2+cations, forming a chain of [Fe(CN)4Cd(en)] units running parallel to the a axis. Along the b axis, adjacent chains are then inter\u00adconnected through the N atoms of the bridging en ligands at the Cd atoms into a two-dimensional layer of [Fe(CN)4Cd(en)2], as shown in Fig.\u00a032}2{Cd(en)}2] units with an Fe\u22efCd distance through the cyanide bridge and a Cd\u22efCd distance through the en bridge of 5.1292\u2005(7) and 7.6692\u2005(12)\u2005\u00c5, respectively. The M\u22efM distances across the cyclic windows vary from 5.5614\u2005(10) to 14.0061\u2005(10)\u2005\u00c5.Compound (II)4]\u2212 mol\u00adecules are linked together through the parallel pyridyl rings of the 2,2\u2032-bipy ligands to generate a graphite-like layers parallel to the ab plane. Within the sheets, the neighbouring pyridyl moieties related by an inversion centre are in a head-to-head arrangement with centroid (Cg) to centroid distances of 4.005\u2005(3)\u2005\u00c5 [inter\u00adplanar angle = 0.0\u2005(4)\u00b0] and 3.903\u2005(3)\u2005\u00c5 [inter\u00adplanar angle = 0.0\u2005(3)\u00b0] for rings A\u22efAi and B\u22efBii , respectively. The FeIII\u22efFeIII separations along the \u03c0\u2013\u03c0 stacking of parallel rings A\u22efAi and rings B\u22efBii are 8.2821\u2005(12) and 8.4572\u2005(13)\u2005\u00c5, respectively. The adjacent pyridyl rings A and Biii related by translation parallel to the a axis are arranged alternately in a head-to-tail manner with a Cg\u22efCg distance of 3.865\u2005(2)\u2005\u00c5 [inter\u00adplanar angle = 1.51\u2005(12)\u00b0] and an FeIII\u22efFeIII separation of 6.8690\u2005(9)\u2005\u00c5.The three-dimensional supra\u00admolecular structure in (I)2O)4 and hexa\u00admeric (H2O)6 subunits into (H2O)10 units [the dihedral angle between the best plane of the (H2O)4 and (H2O)6 subunits is 55.2\u2005(2)\u00b0]; neighbouring units are further joined together, giving rise to ladder-like water chains running parallel to the a axis. As can be seen from Fig.\u00a05i, O2, and O2i (for symmetry code see Table\u00a012O)4 3(DMF)4(H2O)2]\u00b74DMF\u00b74H2O i, O1ii, O2, O2iii, O3, and O3iii ]+ cations lie above and below the water chains and take part in the formation of weak C\u2014H\u22efO hydrogen bonds with the water mol\u00adecule.The hexa\u00admeric water unit has crystallographically imposed inversion symmetry. The six water mol\u00adecules O1via \u03c0\u2013\u03c0 stacking between adjacent pyridyl rings with Cg\u22efCg distances of 4.2925\u2005(18) [inter\u00adplanar angle = 1.55\u2005(18)\u00b0] and 4.0642\u2005(18)\u2005\u00c5 [inter\u00adplanar angle = 0.0\u2005(3)\u00b0] for rings C\u22efCiv and C\u22efCv , respectively, Fig.\u00a06For (II)3)4[Fe(CN)4]\u00b73H2O (I)4[Fe(CN)4]\u00b7H2O . Analysis calculated for C18H26FeN7O: C, 48.66; H, 5.90; N, 22.07%. Found: C, 48.66; H, 5.90; N, 22.07%.The building block N(CH3)2\u00b74H2O and ethyl\u00adenedi\u00adamine were dissolved in distilled H2O (4\u2005ml), and this was pipetted into one side of an H-tube. N(CH3)4[Fe(CN)4]\u00b73H2O was dissolved in distilled H2O (4\u2005ml), and this was pipetted into the other side arm of the H-tube. The H-tube (15\u2005ml capacity) was then carefully filled with distilled H2O. Slow diffusion in the dark for three weeks yielded dark-yellow plate-shaped crystals of (II)18H26CdFeN10O: C, 38.15; H, 4.62; N, 24.72%. Found: C, 38.18; H, 4.60; N, 24.68%.For the synthesis of (II)Uiso(H) = 1.5Ueq(C) for methyl groups and 1.2Ueq otherwise. For (I)Uiso(H) = 1.5Ueq(O). For (II)Uiso(H) = 1.5Ueq(O). The tetra\u00admetyl\u00adammonium cation in (I)A, C17A and C18A, and 0.560\u2005(6) for atoms C16B, C17B, and C18B. Anisotropic displacement parameters of all atoms were restrained using enhanced rigid-bond restraints I, II. DOI: 10.1107/S2056989016006848/bg2584Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016006848/bg2584IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1476008, 1476007CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structures of two bromo\u2013hy\u00addroxy\u2013benzoic acid derivatives, namely, methyl 4-bromo-2-(meth\u00adoxy\u00admeth\u00adoxy)benzoate, (I), and 4-bromo-3-(meth\u00adoxy\u00admeth\u00adoxy)benzoic acid, (II), are compared. Compound (II) crystallizes with two independent mol\u00adecules in the asymmetric unit. In the crystal structures of both compounds, two-dimensional architectures are formed principally by C\u2014H\u22efO hydrogen bonds, and by Br\u22efO inter\u00adactions in (I) and by \u03c0\u2013\u03c0 inter\u00adactions in (II). 10H11BrO4, (I), and C9H9BrO4, (II), are derivatives of bromo\u2013hy\u00addroxy\u2013benzoic acids. Compound (II) crystallizes with two independent mol\u00adecules (A and B) in the asymmetric unit. In both (I) and (II), the O\u2014CH2\u2014O\u2014CH3 side chain is not in its fully extended conformation; the O\u2014C\u2014O\u2014C torsion angle is 67.3\u2005(3) \u00b0 in (I), and \u221265.8\u2005(3) and \u221274.1\u2005(3)\u00b0 in mol\u00adecules A and B, respectively, in compound (II). In the crystal of (I), mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, forming C(5) chains along [010]. The chains are linked by short Br\u22efO contacts [3.047\u2005(2)\u2005\u00c5], forming sheets parallel to the bc plane. The sheets are linked via C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional architecture. In the crystal of (II), mol\u00adecules A and B are linked to form R22(8) dimers via two strong O\u2014H\u22efO hydrogen bonds. These dimers are linked into \u22efA\u2013B\u22efA\u2013B\u22efA\u2013B\u22ef [C22(15)] chains along [011] by C\u2014H\u22efO hydrogen bonds. The chains are linked by slipped parallel \u03c0\u2013\u03c0 inter\u00adactions [inter-centroid distances = 3.6787\u2005(18) and 3.8431\u2005(17)\u2005\u00c5], leading to the formation of slabs parallel to the bc plane.The title compounds, C The dihedral angle between the benzene ring and the ester segment (O1/C7/O2/C8) is 14.5\u2005(2)\u00b0, while the plane through atoms C10/O4/C9 of the meth\u00adoxy\u00admeth\u00adoxy side chain is inclined to the benzene ring by 82.5\u2005(3)\u00b0.The mol\u00adecular structure of compound (I)A and B) in the asymmetric unit. The conformations of the two mol\u00adecules differ in the torsion angles of the \u2013O\u2013CH2\u2013O\u2013CH3 side chains and the orientation of the \u2013COO\u2013 group with respect to the benzene ring, as shown in the AutoMolFit diagram \u00b0, and torsion angle O3B\u2014C8B\u2014O4B\u2013-C9B in mol\u00adecule B is \u221274.1\u2005(3)\u00b0. The dihedral angle between the benzene ring and the plane through atoms C8A/O4A/C9A of the meth\u00adoxy\u00admeth\u00adoxy side chain in mol\u00adecule A is 79.2\u2005(3)\u00b0, while the corresponding dihedral angle in mol\u00adecule B, between the benzene ring and plane C9B/O4B/O8B is 67.1\u2005(3)\u00b0. This is less than in compound (I)A and 9.1\u2005(4)\u00b0 in B; also less than observed in compound (I)viz. 14.5\u2005(2)\u00b0.The mol\u00adecular structure of compound (II)am Fig.\u00a03. The \u2013O\u2013A\u22efO1 hydrogen bonds (Table\u00a01C(5) chains along the b axis. Adjacent chains are linked by short Br1\u22efO4i contacts leading to the formation of sheets parallel to plane (100). The sheets are linked by C5\u2014H5\u22ef\u03c0 inter\u00adactions (centroid of the benzene ring C1\u2013C6) along the a-axis direction, forming a three-dimensional structure s Table\u00a01, forminge Table\u00a01.A and B are linked via two strong O\u2014H\u22efO hydrogen bonds, namely, O2A\u2013-H2A\u22efO1B and O2B\u2013-H2B\u22efO1A, forming dimers with an B\u2014H8B2\u22ef\u00b7O3A hydrogen bonds \u2005\u00c5; Cg2 is the centroid of ring C1B\u2013C6B; inter-planar distance = 3.3691\u2005(12)\u2005\u00c5; slippage = 1.477\u2005\u00c5; symmetry code (ii): \u2212x, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01], and between A and B mol\u00adecules , thus forming slabs lying parallel to the bc plane f Table\u00a02. Adjacens Table\u00a02, formingne Fig.\u00a07.Synthesis of methyl 4-bromo-2-(meth\u00adoxy\u00admeth\u00adoxy) benzoate (I)N,N-diiso\u00adpropyl\u00adethyl\u00adamine (DIPEA), followed by chloro\u00admethyl methyl ether (MOM-Cl), at 273\u2005K and the reaction mixture was stirred at room temperature overnight. The reaction mixture was then diluted with water (50\u2005ml) and the organic layer was extracted with ethyl acetate (2 \u00d7 50\u2005ml). The combined organic layers were washed successively with water, brine, dried over anhydrous magnesium sulfate (MgSO4), filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography using ethyl acetate:hexane (1:9) as eluent to afford (I)1H NMR : \u03b4 = 3.39 , 3.79 , 5.29 7.29 , 7.44 , 7.60 .To a stirred solution of methyl 4-bromo-2-hy\u00addroxy-benzoate in di\u00adchloro\u00admethane (15\u2005ml) (DCM) was added Synthesis of 4-bromo-3-(meth\u00adoxy\u00admeth\u00adoxy)benzoic acid (II)N hydro\u00adchloric acid. The precipitated solid was filtered, dried under vacuum to afford (II)1H NMR : \u03b4 = 3.39 , 5.28 , 7.26 , 7.40 , 7.59 , 12.90 .A mixture of methyl 4-bromo-3-(meth\u00adoxy\u00admeth\u00adoxy) benzoate , 10% aqueous potassium hydroxide , tetra\u00adhydro\u00adfuran (5\u2005ml) and methanol (20\u2005ml) was stirred at room temperature for 2\u2005h. The mixture was then concentrated to remove organic solvents and the aqueous layer was acidified with 6\u2005Single crystals of compounds (I)Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. In the final cycles of refinement reflection (0 0 2) in (I)F2obs and F2calc, considerably improving the values of R1, wR2, and GOF.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016003777/su5284sup1.cifCrystal structure: contains datablock(s) I, II, Global. DOI: Click here for additional data file.10.1107/S2056989016003777/su5284Isup2.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016003777/su5284IIsup3.cmlSupporting information file. DOI: 1457944, 1457943CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The triflate ligand was found to be disordered over two sets of sites, with a site-occupancy ratio of 0.622\u2005(16):0.378\u2005(16). Weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efF hydrogen-bonding inter\u00adactions generate a two-dimensional supra\u00admolecular structure lying parallel to (100). This is only the second crystal structure reported of a mononuclear bis\u00ad(acetyl\u00adacetonato)iron(III) complex.The mononuclear title complex, [Fe(CF The dinuclear alkoxides, [Fe(acac)2(\u03bc-OR)] are also known iron(III) complex, [Fe(acac)2(OTf)(THF)], the title compound (I)Because of its ease-of-handling, relative stability and good solubility in most organic solvents, tris\u00ad(acetyl\u00adacetonato)iron(III) [Fe(acac)3+ atom in an slightly distorted octa\u00adhedral FeO6 environment. The coordination sphere of the metal comprises four oxygen atoms from two \u03ba2-acac ligands [Fe\u2014Oacac range = 1.9517\u2005(11)\u20131.9762\u2005(11)\u2005\u00c5], one oxygen atom of a THF solvate mol\u00adecule [Fe1\u2014O8 = 2.0781\u2005(11)\u2005\u00c5] and one oxygen atom of a disordered triflate anion [Fe1\u2014O5 = 2.063\u2005(4)\u2005\u00c5 or Fe1\u2014O5B = 2.066\u2005(6)\u2005\u00c5] (Table\u00a01cis angles range from 84.63\u2005(5)\u00b0 to 98.09\u2005(5)\u00b0 and the trans angles range from 172.60\u2005(5)\u00b0 to 174.9\u2005(6)\u00b0.The mol\u00adecular structure of the mononuclear complex (I)] Table\u00a01. The disa,b). A series of Cmeth\u00adyl\u2014H\u22efOacac, Cmeth\u00adyl\u2014H\u22efOtriflate, Cmeth\u00adyl\u2014H\u22efFtriflate, CTHF\u2014H\u22efOtriflate, and CTHF\u2014H\u22efFtriflate inter\u00adactions make up the layers, the details of these inter\u00adactions are presented in Table\u00a02c,d.There are no significant supra\u00admolecular features to discuss in the extended structure of (I)2Cl] acac range of 1.945\u20132.062\u2005\u00c5.Only one other mononuclear bis\u00ad(acetyl\u00adacetonato)iron(III) complex has been characterized crystallographically, [Fe(acac)2(C6H5)(OTf)] (THF)] -triflate complex, [Os(acac)3] in dry THF (5\u2005mL). The resulting purple\u2013red solution was stirred at room temperature for 1\u2005h. The reaction mixture was then concentrated under vacuum to a volume of approximately 2\u2005mL, and 20\u2005mL of pentane was added. A dark purple\u2013red microcrystalline solid precipitated. The mixture was filtered through a glass-frit and the microcrystalline solid was dried under vacuum . Crystals suitable for X-ray diffraction were grown by slow diffusion of pentane into a THF solution of the purple\u2013red solid. CH analysis calculated for C15H22F3FeO8S (MW: 475.235): C 37.91%; H 4.67%. Found: C 37.69%; H, 4.45%.Triflic acid was added to a solution of . Remaining H atoms were also included as riding idealized contributors .Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015016849/zs2341sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015016849/zs2341Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015016849/zs2341Isup3.tifSupporting information file. DOI: 1423096CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports6: Article number: 2427810.1038/srep24278; published online: 04072016; updated: 06102016.This Article contains an error in Figure S3; where the labels \u20180\u2009\u00b0C\u2019 and \u201825\u2009\u00b0C\u2019 are inverted. The correct Figure S3 appears below as"} +{"text": "In the crystal, mol\u00adecules are linked 12H13N2+\u00b7Cl\u2212, the chloride salt of 1,1\u2032-di\u00adphenyl\u00adhydrazine, the phenyl rings are inclined to one another by 78.63\u2005(17)\u00b0. The N\u2014+NH3 bond lengths is 1.445\u2005(3)\u2005\u00c5, and the N\u2014Cphen\u00adyl bond lengths are 1.435\u2005(3) and 1.447\u2005(4)\u2005\u00c5. In the crystal, mol\u00adecules are linked via N\u2014H\u22efCl hydrogen bonds, forming chains along [10-1], which enclose two adjacent R42(6) ring motifs. The chains are reinforced by C\u2014H\u22efCl hydrogen bonds.In the title compound, C It has been used as a starting reagent for the preparation of Schiff bases as fluorescent sensors for fluoride . The latter compound is one of a series that has been used to prepare bis\u00ad(imino)\u00adpyridyl iron and cobalt complexes to study the effect of nitro\u00adgen substituents on ethyl\u00adene oligomerization and polymerization via N\u2014H\u22efCl hydrogen bonds, forming chains along . There are no \u03c0\u2013\u03c0 inter\u00adactions present in the crystal structures of any of the three compounds.In the crystal of compound (II), mol\u00adecules are linked by N\u2014H\u22efN, N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds, forming two-dimensional networks parallel to (001). These sheets are linked N,N-di\u00adphenyl\u00adhydrazone) by a condensation reaction involving 1,1\u2032-di\u00adphenyl\u00adhydrazinium hydro\u00adchloride and 2,6-di\u00adacetyl\u00adpyridine in methanol.Brown block-like crystals of the title compound were obtained during an attempt to prepare the ligand 2,6-di\u00adacetyl\u00adpyridine bis\u00ad(Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S1600536814022879/lh5735sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S1600536814022879/lh5735Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814022879/lh5735Isup3.cmlSupporting information file. DOI: 1029761CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "However, the crystal structure exhibits pseudosymmetry as the two independent mol\u00adecules are related by an approximate but non-crystallographic inversion located close to in the selected asymmetric unit, and the structure exhibits partial inversion twinning. The approximate inversion relationship between the two mol\u00adecules in the selected asymmetric unit is clearly shown by comparison of the relevant torsion angle in the two mol\u00adecules; the corresponding torsion angles have similar, although not identical magnitudes but with opposite signs. The mean planes of the quinoline rings in the two independent mol\u00adecules are almost parallel, with a dihedral angle of only 0.16\u2005(3)\u00b0 between them, and the mutual orientation of these rings permits significant \u03c0\u2013\u03c0 stacking inter\u00adactions between them [centroid\u2013centroid distances = 3.7579\u2005(15) and 3.7923\u2005(15)\u2005\u00c5]. In addition, the bimolecular aggregates which are related by translation along [010] are linked by a further \u03c0\u2013\u03c0 stacking inter\u00adaction [centroid\u2013centroid distance = 3.7898\u2005(15)\u2005\u00c5], so forming a \u03c0-stacked chain running parallel to [010]. However, there are no C\u2014H\u22efN hydrogen bonds in the structure nor, despite the number of independent aromatic rings, are there any C\u2014H\u22ef\u03c0 hydrogen bonds; hence there are no direction-specific inter\u00adactions between adjacent \u03c0-stacked chains.The title compound, C \u00c5b = 7.1401 (3) \u00c5c = 13.0804 (5) \u00c5V = 2532.47 (17) \u00c53Z = 8K\u03b1 radiationMo \u22121\u03bc = 0.30 mmT = 173 K0.48 \u00d7 0.32 \u00d7 0.22 mmAgilent Eos Gemini diffractometerCrysAlis RED; Agilent, 2012Tmin = 0.808, Tmax = 0.936Absorption correction: multi-scan (29727 measured reflections5975 independent reflectionsI > 2\u03c3(I)5204 reflections with Rint = 0.037R[F2 > 2\u03c3(F2)] = 0.040wR(F2) = 0.097S = 1.085975 reflections331 parameters1 restraintH-atom parameters constrainedmax = 0.25 e \u00c5\u22123\u0394\u03c1min = \u22120.22 e \u00c5\u22123\u0394\u03c1x determined using 1610 quotients [(I+)\u2212(I\u2212)]/[(I+)+(I\u2212)] CrysAlis PRO global, I. DOI: 10.1107/S205698901500804X/hg5440Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901500804X/hg5440Isup3.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901500804X/hg5440fig1.tif. DOI: The two independent mol\u00adecules in the title compound showing the atom-labelling scheme. Displacement ellipsoids are shown at the 30% probability level.Click here for additional data file.10.1107/S205698901500804X/hg5440fig2.tif. DOI: The two mol\u00adecules in the selected asymmetric unit, viewed normal to the planes of the quinolone units, showing the ring overlap which leads to a \u03c0..\u03c0 sktacking inter\u00adaction. For the sake of clarity, the H atoms have been omitted.Click here for additional data file.10.1107/S205698901500804X/hg5440fig3.tif. DOI: A stereoview of part of the crystal structure of the title compound showing the formation of a \u03c0-stacked chain parallel to [010]. For the sake of clarity, the H atoms have been omitted.1061227CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The bond lengths and angles of the cyclo\u00adpenta\u00addienyl and phosphane ligands are very similar to that of the unsolvated complex [Taqui Khan et al. and 1.156\u2005(3)\u2005\u00c5 and has an N\u2014N\u2014Ru angle of 119.20\u2005(15)\u00b0, indicating a greater contribution of the canonical form Ru\u2014N=N(+)=N(-) for the bonding situation. An intra\u00admolecular C\u2014H\u22efN hydrogen-bonding inter\u00adaction between one ortho H atom of a phosphane ligand and the N atom coordinating to the metal is observed. A similar inter\u00admolecular inter\u00adaction is observed between a meta H atom of a phosphane ligand and the terminal azide N atom of a neighbouring complex. Finally, two C\u2014H\u22efN inter\u00adactions exists between the H atoms of the di\u00adchloro\u00admethane solvent mol\u00adecule and the terminal N atom of two azide anions. The solvent mol\u00adecule is located about a twofold rotation axis and shows disorder of the Cl atoms with an occupancy ratio of 0.62\u2005(3):0.38\u2005(3).The title solvated complex, [Ru(\u03b7 al. 1994. Acta Cr For oth al. 1999; Govinda al. 2005. For met al. 2005; Seok & al. 2005. Non-cla al. 2005 of respe al. 2002.5H5)(N3)(C18H15P)2]\u00b70.5CH2Cl2[Ru \u00c5b = 12.4559 (3) \u00c5c = 28.6781 (6) \u00c5\u03b2 = 94.213 (2)\u00b0V = 7189.7 (3) \u00c53Z = 8K\u03b1 radiationMo \u22121\u03bc = 0.63 mmT = 293 K0.72 \u00d7 0.51 \u00d7 0.20 mmAgilent Xcalibur Atlas Gemini diffractometerCrysAlis PRO 6038 reflections with Rint = 0.032R[F2 > 2\u03c3(F2)] = 0.027wR(F2) = 0.065S = 1.067106 reflections449 parametersH-atom parameters constrainedmax = 0.33 e \u00c5\u22123\u0394\u03c1min = \u22120.49 e \u00c5\u22123\u0394\u03c1CrysAlis PRO I, New_Global_Publ_Block. DOI: 10.1107/S1600536814019187/wm5050Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814019187/wm5050fig1.tif. DOI: Mol\u00adecular structure of the title compound showing the atom-numbering scheme. Displacement ellipsoids are drawn at the 30% probability level. Only the major component of the disordered di\u00adchloro\u00admethane solvate is shown. Hydrogen atoms of the metal complex have been removed for clarity.Click here for additional data file.10.1107/S1600536814019187/wm5050fig2.tif. DOI: View of the mol\u00adecular arrangement in the title structure viewed along [010]. Hydrogen bonds are denoted by dashed lines.1021189CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atom with a distorted octa\u00adhedral coordination environment defined by six N atoms from three bidentate phenanthroline ligands. The non-coordinating N,N\u2032-diglycine ligand links the cationic building blocks via C\u2014H\u22efO contacts and through lone-pair\u22ef\u03c0 inter\u00adactions. Further observed non-covalent inter\u00adactions contribute to the consolidation of the supra\u00admolecular network.The complex cation of the title compound includes one Co 12H8N2)3]2(NO3)4\u00b7C12H12N2O6\u00b78H2O, contains a CoII atom with a distorted octa\u00adhedral coordination environment defined by six N atoms from three bidentate 1,10-phenanthroline ligands. The asymmetric unit of the title compound is completed by one-half of the N,N\u2032-diglycine solvent mol\u00adecule, which is located on a centre of inversion, by two nitrate counter-anions and four solvent water mol\u00adecules. Two [Co(C12H8N2)3]2+ cations are connected through C\u2014H\u22efO contacts and through lone-pair\u22ef\u03c0 inter\u00adactions involving the non-coordinating N,N\u2032-diglycine and phenanthroline mol\u00adecules. The different aromatic ring systems are involved in \u03c0\u2013\u03c0 stacking and C\u2014H\u22ef\u03c0 inter\u00adactions, with centroid-to-centroid distances in the range 3.7094\u2005(8)\u20133.9973\u2005(9)\u2005\u00c5. The crystal structure is stabilized by further anion\u22ef\u03c0 inter\u00adactions and C\u2014H\u22efO contacts, as well as O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds between water mol\u00adecules, the non-coordinating nitrate anions, N,N\u2032-diglycine and phenanthroline mol\u00adecules. These non-covalent inter\u00adactions give rise to a three-dimensional supra\u00admolecular network.The complex cation of the title compound, [Co(C Application fields for these materials are in catalysis, in gas storage diglycine as metal-linking ligand, zigzag chains are formed, constructing inter\u00adpenetrating networks (see Database survey). In our synthetic approach, we offer such systems another electron-deficient bidentate aromatic ring system like phenanthroline or bi\u00adpyridine in order to block parts of the coordination sphere of the metal atoms so that these zigzag chains are truncated or not formed at all. Thus, an alternative route for the resultant system lies in the use of the offered \u03c0-inter\u00adaction possibilities as well as in stacking inter\u00adactions as a new linking mode. Recently, we have described the inter\u00adactions of a cobalt(III) bi\u00adpyridine complex with supra\u00admolecular synthons diglycine. The chosen ligand N,N\u2032-diglycine is a relatively rigid mol\u00adecule with one sp3-hybridized methyl\u00adene carbon atom that allows the acid moiety to rotate. Moreover, this ligand simultaneously possesses several coordination sites through the carb\u00adoxy\u00adlic group and the oxygen atom of the amide group. These functional groups can also be involved in hydrogen bonding and D\u2014H\u22ef\u03c0 inter\u00adactions.In previously synthesized transition metal complexes with N,N\u2032-diglycine solvent molecule linking two tris\u00ad(phenanthroline)cobalt(II) cationic building blocks via the mentioned non-classical inter\u00adactions.In the present contribution we have determined the structure of a novel cobalt(II) coordination polymer with a non-coordinating II complex cation in which three bidentate phenanthroline ligands define a distorted octa\u00adhedral coordination sphere. Distances and angles of this rather common cationic species, [Co(C12H8N2)3]2+, are well within expected ranges and are comparable to those found in the literature diglycine molecule. The asymmetric unit is completed by two non-coordinating nitrate counter-anions and four solvent water mol\u00adecules. The N,N\u2032-diglycine mol\u00adecule links two complex tris\u00adcobalt(II) cations via lone-pair\u22ef\u03c0 inter\u00adactions involving the carb\u00adoxy\u00adlic acid function and the phenanthroline aromatic system as well as C\u2014H\u22efO contacts between the oxygen atom of the amide group and one phenanthroline ligand. Moreover, \u03c0\u2013\u03c0 stacking inter\u00adactions between different aromatic ring systems and C\u2014H\u22ef\u03c0 as well as O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonding are observed and consolidate an extensive three-dimensional supra\u00admolecular network.The mol\u00adecular entities Fig.\u00a01 of the tvia O\u2014H\u22efO, C\u2014H\u22efO and partly via N\u2014H\u22efO hydrogen bonds with water, phenanthroline and N,N\u2032-diglycine mol\u00adecules diglycine solvent mol\u00adecules stack these components along the c axis \u2005\u00c5 and between Cg7\u22efCg7 is 3.9973\u2005(9)\u2005\u00c5 \u22efCg8 of 3.037\u2005(1)\u2005\u00c5, where Cg8 is the centroid defined by the ring atoms N5/C25\u2013C28/C36. Moreover, a relatively short N\u2014H\u22ef\u03c0 distance of 4.08\u2005(6)\u2005\u00c5 is observed diglycine solvent and the Cg2 centroid of a phenanthroline ligand are associated with a distance of 3.400\u2005(5)\u2005\u00c5. Similar distances of 3.461\u2005(5)\u2005\u00c5 prevail between the O10 atom of a water mol\u00adecule and the Cg3 centroid of a phenanthroline ligand, where Cg2 and Cg3 are the centroids defined by the ring atoms N1/C1\u2013C4/C12 and C4\u2013C7/C11/C12, respectively. The values are similar to those found in the literature \u2005\u00c5 that is comparable to previously reported structures diglycine resulted in six metal-organic compounds diglycine and shows a number of non-classical inter\u00adactions diglycine, was prepared by the method of Cleaver & Pratt (1955v/v) mixture of water and methanol (50\u2005ml) and refluxed for 30 minutes. The mixture was allowed to cool to room temperature, and a previously prepared aqueous solution of cobalt nitrate (1\u2005mmol) was slowly added under continuous stirring. Deep dark-orange block-shaped crystals of the title compound were obtained by slow evaporation at room temperature.The starting material, ratt 1955. Cesium Uiso(H) = 1.2 Ueq(C) and C\u2014H(aromatic) = 0.94\u2005\u00c5 and C\u2014H(methyl\u00adene) = 0.98\u2005\u00c5 using a riding model. The water H atoms were located in a different Fourier map and were refined with O\u2014H distances restrained to 0.82\u20130.87\u2005\u00c5 and with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015013006/wm5178sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989015013006/wm5178Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015013006/wm5178Isup4.cdxSupporting information file. DOI: 1410901CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I with the pyridyl\u00adimidazol ligand 4--1H-imidazol-2-yl)pyridine, afforded a nitrate-free one-dimensional looped-chain polymeric structure. The AgI cation adopts a highly distorted tetra\u00adhedral geometry coordinated by two pyridine N atoms and two imidazole N atoms from four individual ligands.The reaction of Ag 29H25N3)2]NO3\u00b7CH3OH\u00b7H2O}n, the AgI cation is four-coordinated by two pyridine N atoms and two imidazole N atoms from four individual 4--1H-imidazol-2-yl)pyridine (i-pro-pyim) ligands. This gives rise to a highly distorted tetra\u00adhedral geometry with bond angles falling in the range 100.33\u2005(19)\u2013122.76\u2005(19)\u00b0. Two crystallographically independent i-pro-pyim ligands (A and B) adopt very similar conformations to one another, such that the dihedral angles between the pyridyl and imidazolyl rings in the two ligands are 40.7\u2005(3) and 42.2\u2005(3)\u00b0, respectively. Each i-pro-pyim ligand binds two symmetry-related Ag+ cations, leading to the formation of 14-membered cyclic dimers, in which the AgI atoms are separated by 6.963\u2005(2)\u2005\u00c5 for the Ag\u2013A2\u2013Ag dimer and 7.020\u2005(2)\u2005\u00c5 for Ag\u2013B2\u2013Ag. These cyclic dimers are alternately connected to each other by sharing AgI atoms, resulting in the formation of a looped-chain structure extending along the [100] direction. Moreover, adjacent looped chains are connected by inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid-to-centroid distance = 3.689\u2005(4)\u2005\u00c5], giving rise to the formation of a two-dimensional supra\u00admolecular network propagating parallel to (110). Several inter\u00admolecular C\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds further contribute to the stabilization of the crystal structure.In the title compound, {[Ag(C The coordination polymer is obtained by addition of the ligand to AgNO3 in methanol/aceto\u00adnitrile. The title nitrate salt is closely related to the perchlorate salt because of their high efficiency and long-term stability , one nitrate anion, one methanol solvent mol\u00adecule, and two water solvent mol\u00adecules, each with an occupancy factor of 0.5, in the asymmetric unit. As shown in Fig.\u00a01I atom is coordinated by two pyridine N atoms and two imidazole N atoms from four individual i-pro-pyim ligands, giving rise to a highly distorted tetra\u00adhedral geometry with bond angles falling in the range of 100.33\u2005(19)\u2013122.76\u2005(19)\u00b0 and 42.2\u2005(3)\u00b0, respectively. Moreover, there are intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the pyridyl and phenyl rings of both ligand types, N3,C4\u2013C8 and C21\u2013C26 [centroid-to-centroid distance = 3.760\u2005(4)\u2005\u00c5] for A and N6,C33\u2013C37 and C51\u2013C56 [centroid-to-centroid distance = 3.716\u2005(4)\u2005\u00c5] for B.In the title compound there are two crystallographically independent ligands, A ligands link two AgI atoms, resulting in the formation of a 14-membered cyclic dimer with an Ag\u22efAg distance of 6.963\u2005(2)\u2005\u00c5 and a \u03c0\u2013\u03c0 inter\u00adaction [centroid-to-centroid distance = 3.890\u2005(4)\u2005\u00c5] between N3-containing pyridine rings \u2005\u00c5 and a \u03c0\u2013\u03c0 inter\u00adaction [centroid-to-centroid distance = 3.922\u2005(4)\u2005\u00c5] between N6-containing pyridine rings. The two cyclic dimers are connected alternately by sharing AgI atoms, leading to the formation of a looped-chain structure extending along the a axis \u2212x\u00a0+\u00a02, \u2212y, \u2212z\u00a0+\u00a01], resulting in the formation of a two-dimensional supra\u00admolecular network propagating parallel to (110) \u2005\u00c5] between the C50\u2013C55 and C500) Fig.\u00a03. No nota0) Fig.\u00a03.i-pro-pyim ligand was synthesized according to literature procedures and allowing the solution to evaporate slowly at room temperature.The W and O2W) were refined with site-occupancy factors of 0.5, and their H atoms were not included in the model. All H atoms except those of the water mol\u00adecules were positioned geometrically and refined using a riding model, with d(C\u2014H) = 0.95\u2005\u00c5 for Csp2\u2014H, 1.00\u2005\u00c5 for methine, C\u2014H, 0.98\u2005\u00c5 for methyl, and O\u2014H 0.84\u2005\u00c5 for hydroxyl H atoms. For all H atoms, Uiso(H) = 1.2\u20131.5Ueq of the parent atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901600949X/sj5501sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S205698901600949X/sj5501Isup2.hklStructure factors: contains datablock(s) I. DOI: 1484735CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title salt, crystalline water mol\u00adecules serve as donors for the weak inter\u00admolecular O\u2014H\u22efO and O\u2014H\u22efBr hydrogen bonds which link adjacent polymeric chains. 2(C5H9NO2)]\u00b7H2O}n, the CdII ion is coordinated by four bromido ligands and two carboxyl\u00adate oxygen atoms of two symmetry-related proline ligands, which exist in a zwitterionic form, in a distorted octa\u00adhedral geometry. There is an intra\u00admolecular N\u2014H\u22efO hydrogen bond between the amino group and the carboxyl\u00adate fragment. Each coordinating ligand bridges two CdII atoms, thus forming polymeric chains running along the c-axis direction. The water mol\u00adecules of crystallization serve as donors for the weak inter\u00admolecular O\u2014H\u22efO and O\u2014H\u22efBr hydrogen bonds that link adjacent polymeric chains, thus forming a three-dimensional structure. N\u2014H\u22efO and N\u2014H\u22efBr hydrogen bonds also occur.In the title coordination polymer, {[CdBr It is known that the chiral amino acids and their complexes are potential materials for NLO applications \u20132.7737\u2005(12)\u2005\u00c5] and two carboxyl\u00adate oxygen atoms [Cd\u2014O = 2.312\u2005(8) and 2.318\u2005(8)\u2005\u00c5] of two proline ligands in a slightly distorted octa\u00adhedral geometry. The title complex is extended as a polymeric chain which runs parallel to the c axis. Within one chain, adjacent CdII ions are separated by 3.727\u2005(1)\u2005\u00c5. The closest Cd\u22efCd distance between neighbouring polymeric chains is 8.579\u2005(2)\u2005\u00c5. The five endocyclic torsion angles of the pyrrolidine ring of the proline residue are N1\u2014C2\u2014C3\u2014C4 = 31.8\u2005(13)\u00b0, C2\u2014C3\u2014C4\u2014C5 = \u221239.1\u2005(15)\u00b0, C3\u2014C4\u2014C5\u2014N1 = 29.9\u2005(14)\u00b0, C2\u2014N1\u2014C5\u2014C4 = \u22129.7\u2005(12)\u00b0 and C5\u2014N1\u2014C2\u2014C3 = \u221213.1\u2005(11)\u00b0. The pyrrolidine ring exhibits twisted conformation on the C3\u2014C4 bond with a pseudo-rotation angle \u0394 = 249.3\u2005(12)\u00b0 and a maximum torsion angle \u03d5m = 38.5\u2005(8)\u00b0 et al., 1983A\u22efO2 hydrogen bond between the amino group and the carboxyl\u00adate fragment.In (I)The crystal structure of (I)viz. catena-[di\u00adchlorido-(4-hy\u00addroxy-l-proline)cadmium] (\u03bc2-l-pro\u00adline)cadmium monohydrate] manganese(II) monohydrate], has been structurally determined three times and has similar cell parameters and the same space group as the title compound and cadmium bromide tetra\u00adhydrate (Loba) in an equimolar ratio were dissolved in double-distilled water. The obtained solution of the homogeneous mixture was evaporated at room temperature to afford the white crystalline title compound, which was then recrystallized by slow evaporation from an aqueous solution.To prepare the title compound, et al., 1983W\u22efH2W distances of the water mol\u00adecules were restrained to 0.85\u2005(2) and 1.38\u2005(2)\u2005\u00c5, respectively, using the DFIX option and included in the structure-factor calculations with Uiso(H1W/H2W) = 1.1Ueq(O1W). The remaining hydrogen atoms were placed in geometrically idealized positions (C\u2014H = 0.97\u20130.98\u2005\u00c5 and N\u2014H = 0.89\u2005\u00c5) with Uiso(H) = 1.2Ueq(C/N) and were constrained to ride on their parent atoms. Reflections 110 and 020 were partially obscured by the beam stop and were omitted.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015001176/cv5483sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015001176/cv5483Isup2.hklStructure factors: contains datablock(s) I. DOI: 1044327CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Stevens classification of levels of measurement involves four types of scale: \u201cNominal\u201d, \u201cOrdinal\u201d, \u201cInterval\u201d and \u201cRatio\u201d. This classification has been used widely in medical fields and has accomplished an important role in composition and interpretation of scale. With this classification, levels of measurements appear organized and validated. However, a group theory-like systematization beckons as an alternative because of its logical consistency and unexceptional applicability in the natural sciences but which may offer great advantages in clinical medicine. According to this viewpoint, the Stevens classification is reformulated within an abstract algebra-like scheme; \u2018Abelian modulo additive group\u2019 for \u201cOrdinal scale\u201d accompanied with \u2018zero\u2019, \u2018Abelian additive group\u2019 for \u201cInterval scale\u201d, and \u2018field\u2019 for \u201cRatio scale\u201d. Furthermore, a vector-like display arranges a mixture of schemes describing the assessment of patient states. With this vector-like notation, data-mining and data-set combination is possible on a higher abstract structure level based upon a hierarchical-cluster form. Using simple examples, we show that operations acting on the corresponding mixed schemes of this display allow for a sophisticated means of classifying, updating, monitoring, and prognosis, where better data mining/data usage and efficacy is expected. In 1946, S. S. Stevens devised his classification of \u201clevels of measurement\u201d ,3.Thus, in the following, we re-interpret Stevens classification, and endeavour to give it meaning in some abstract algebra-like modelling. There, the most preferred construct is a vector-like structure of various sets of scores based on individual scales and operators that permit changes of score within the set. Additionally, classical datasets that are classified in terms of the Stevens scales of measurement can be mined and combined on a higher abstract structure level based upon a hierarchical-cluster form. To explore this possibility, we provide simple examples to help readers understand this modelling tool.Stevens classified the scales of measurement into four scale types ; \u0406) \u201cNomFor \u0406), the \u201cNominal scale\u201d, there seems to be little room where group theoretical operations apply because within that scale only a labelling scheme is permissible. Although some non-cyclic group might be definable, it seems that little meaning can be attached to operations for this sort of scale.N\u2009=\u2009{0, 1, 2, 3,\u2026, N \u2013 2, N \u2013 1}, where modulo N addition is postulated. With this assumption, given two elements \u2018Xj\u2019 and \u2018Xk\u2019 corresponding for example to the severity of a clinical symptom and/or finding, then composition (denoted by \u2018*\u2019) is taken to be modulo N addition; \u2018Xj*X(j\u2192k)\u2009=\u2009Xk\u2019 (with X(j\u2192k) \u2208 ZN). Here \u2018X(j\u2192k)\u2019 is an operator that produces the change in score, \u2018Xj\u2009\u2192\u2009Xk\u2019 \u2009=\u2009Xj-1*Xk\u2009=\u2009Xk \u2013 Xj\u2019). Then, all scores \u2018Xj\u2019s and operators \u2018X(j\u2192k)\u2019 are composable within a single Abelian modulo additive group \u2018ZN\u2019, where \u2018Xj*Xk\u2009=\u2009Xk*Xj\u2019 holds, at least, in terms of operation \u2018*\u2019. Thus a patient\u2019s state corresponding to a certain illness or disease can be changed through the application of a single operation determined by the two elements belonging to \u2018ZN\u2019 , the combination of all scores being \u20180\u2019 and represented by the identity element for group Y\u2009=\u2009{ZN\u00d7n, *}. Here, Y is the n-fold Cartesian product of \u2018ZN\u2019 (n: the number of components) that comprises all possible assessments related to each state of a given disease, for instance, \u2018hypertension\u2019, \u2018hyperglycaemia\u2019, \u2018diabetes mellitus\u2019, \u2018acute pancreatitis\u2019, \u2018systemic lupus erythematosus\u2019, and \u2018cerebral artery stroke\u2019. If in addition composition is given by modulo \u2018N\u2019 arithmetic, prime numbers are preferable , cj\u2009=\u2009Xj\u2009-\u2009[Xj], \u20180\u2009\u2264\u2009cj\u2009<\u20091\u2019; \u2018[X]\u2019 is the floor function meaning the highest integer below \u2018X\u2019). Similarly,\u2009-\u2009(v) can be redefined using the unit length \u20181\u2019 as an interval scale,0\u2019 (=0) that satisfies \u2018Xj \u25e6X0\u2009=\u2009X0 \u25e6Xj (=Xj\u2009+\u2009X0\u2009=\u2009Xj\u2009+\u20090)\u2009=\u2009Xj\u2019. Additionally, the inverse element is \u2018Xj-1\u2009=\u2009-Xj\u2019 satisfying \u2018Xj-1\u25e6Xj\u2009=\u2009Xj \u25e6Xj-1\u2009=\u2009Xj\u2009+\u2009Xj-1\u2009=\u2009Xj\u2009-\u2009Xj\u2009=\u2009X0 (=0)\u2019.There exists an identity element \u2018Xj\u2019s, i.e., U\u2009\u2261\u2009{Xj | Xj \u2208 R}. Because \u2018Xj , Xk, Xj \u2208 set U, the closure law holds. Therefore, this operation defines a group U\u2009=\u2009{Xj, \u25e6} (mEq/l), sodium: [Na+] (mEq/l), calcium: [Ca++] (mg/dl), chloride: [Cl-] (mEq/l) and bicarbonate: [HCO3-] (mEq/l)\u2019. Also, there are clinical scales whose ranges are the open interval like \u2018-\u221e\u2009<\u2009X\u2009<\u2009+\u221e\u2019 ; \u2018Anion gap [AG]\u2009=\u2009[Na+]\u2009-\u2009([Cl-]\u2009+\u2009[HCO3-]) (reference range for blood tests: 12\u2009\u00b1\u20092\u00a0mEq/l)\u2019 and \u2018Base excess [BE] (reference range for blood tests: 0\u2009\u00b1\u20092\u00a0mmol/l)\u2019. However, both can be treated using the notion of \u2018field\u2019 because those values are real numbers where all four arithmetic operations are included, with the exception of division by zero. Thus, the above clinical values could be definable over a \u2018field\u2019. In this regard, we assume a rule that each unit like \u2018mEq/l\u2019 accompanies the value automatically with the results of operations regardless of types of operation among the four arithmetic operations (Note that there are cases when units vanish as when ratios are taken \u2018mEq/mEq (unitless)\u2019 or displayed in reciprocal form like \u2018l/mEq\u2019). Examples for \u2018[WBC] (/mm3)\u2019, \u2018[Na+] (mEq/l)\u2019 are presented in Appendix C.Nevertheless, other clinical scales range over a semi-open continuous interval like \u20180\u2009\u2264\u2009X\u2009<\u2009+\u221e\u2019 , such as \u2018blood concentration of white blood cells: [WBC] (/mmj # Xk\u2009=\u2009Xl (\u2208V), where ordinal arithmetic calculations are performed excluding of course division by zero.In this case, we consider a set V and assume that \u2018#\u2019 means one of \u2018addition, subtraction, multiplication, and division\u2019 collectively; thus, \u2018Xj\u2009+\u2009Xk\u2009=\u2009Xk\u2009+\u2009Xj, and associative: (Xj\u2009+\u2009Xk)\u2009+\u2009Xl\u2009=\u2009Xj\u2009+\u2009(Xk\u2009+\u2009Xl). As for multiplication, set V meets the conditions of a \u2018monoid\u2019 (/mm3)\u201d, field V2\u2009\u2261\u2009{X2j | X2j \u2208 R (mEq/l)} and V2\u2009=\u2009{X2j, #} for \u201cadministration of a certain drug like lithium carbonate: [Li+] (mEq/l)\u201d, field V3\u2009\u2261\u2009{X3j | X3j \u2208 R (mEq/l)} and V3\u2009=\u2009{X3j, #} for \u201csodium: [Na+] (mEq/l)\u201d, field V4\u2009\u2261\u2009{X4j, #} for calcium: [Ca++] (mg/dl), field V5 for chloride: [Cl-] (mEq/l), field V6 for \u2018Anion gap [AG] (mEq/l)\u2019, field V7 for \u2018Base excess [BE] (mmol/l)\u2019,\u2026, VN,\u2026, . For each, an independent abstract algebraic treatment is possible as for ordinal abstract algebra.Furthermore, different fields based on different sets of clinical values can be described as follows: field Vj\u2019, j\u2009=\u20091, 2, 3,\u2026: number of sessions), where the mixed expression and its totality of operations that could be performed belong to a single set R. Because of the possible variety of operation rules, the genuine use of this set may be unwieldy at this stage.By making use of all types of scales of measurement, we propose a vector-like expression of a patient\u2019s state | clock time for the onset of sleep (/24\u00a0hrs) | blood concentration of white blood cell [WBC] (/mm3) | blood concentration of [Na+] (mEq/l)| a certain value ],Rj\u2019 changes to \u2018Rj+1\u2019 effected by operator \u2018R(j\u2192j+1)\u2019; we denote by \u2018\u25ca\u2019 the binary composition composed of the product of compositions for each component. Three possible states are:Next, suppose the patient\u2019s state \u2018R(1)1\u2019,\u2018X(2)1\u2019, and \u2018X(3)1\u2019, modulo 7 arithmetic (addition) is used. For the 2nd components, \u2018X(1)2, X(2)2, X(3)2\u2019, operations of Abelian addition are used. For the 3rd component, \u2018X(1)3, X(2)3, X(3)3\u2019, 4th \u2018X(1)4, X(2)4, X(3)4\u2019, the four arithmetic operators (those operations denoted by \u2018#\u2019) are required, and for the 5th, \u2018X(1)5, X(2)5, X(3)5\u2019, a certain operational unit is postulated. In the following examples, only addition/subtraction is presented; naturally, multiplication/division is also considered permissible.For the 1st component, \u2018X(1\u2192\u20092)\u2009=\u2009[X(1\u2192\u20092)1(mod\u20097)\u00a0|\u00a0X(1\u2192\u20092)2(/24\u00a0hrs)|\u00a0X(1\u2192\u20092)3(/mm3)|\u00a0X(1\u2192\u20092)4(mEq/l)|X(1\u2192\u20092)5(\u2026)], and R(2\u2192\u20093)\u2009=\u2009[X(2\u2192\u20093)1(mod\u20097)\u00a0|\u00a0X(2\u2192\u20093)2(/24\u00a0hrs)|\u00a0X(2\u2192\u20093)3(/mm3)|\u00a0X(2\u2192\u20093)4(mEq/l)|X(2\u2192\u20093)5(\u2026)]With related operators R(1\u21922)\u2019 and \u2018R(2\u21923)\u2019 from the three states given above are as follows:Then, using results in Appendix D, \u2018RThus, we confirm the relationDetails are illustrated in Appendix E.0)\u2009=\u2009[0 (mod 7)| 0 (/24\u00a0hrs)| 0 (/mm3)| 0 (mEq/l) | X0 (\u2026)]\u2019 such that \u2018Rj\u25caE\u2009=\u2009E\u25caRj\u2009=\u2009Rj\u2019. Additionally, there exists an inverse for any \u2018Rj\u2019, \u2018Rj-\u20091\u2009=\u2009[X(j)1-\u20091(mod\u20097)\u00a0|\u00a0X(j)2-\u20091(/24\u00a0hrs)|\u00a0X(j)3-\u20091(/mm3)|\u00a0X(j)4-\u20091(mEq/l)|X(j)5-\u20091(\u2026)] =\u2009[7\u2013X(j)1(mod\u20097)\u00a0|\u00a024\u2009-\u2009X(j)2(/24\u00a0hrs)|\u2009-\u2009X(j)3(/mm3)|\u2009-\u2009X(j)4(mEq/l)|X(j)5-\u20091(\u2026)]\u2019 that satisfies \u2018Rj-1\u25caRj\u2009=\u2009Rj\u25caRj-1\u2009=\u2009E\u2019. However, commutativity, \u2018Rj\u25caRk\u2009=\u2009Rk\u25caRj\u2019 and associativity, \u2018(Rj\u25caRk)\u25caRl\u2009=\u2009Rj\u25ca(Rk\u25caRl)\u2019 are not satisfied. Here, we assume that operators acting on \u2018Rj\u2019s should be performed from left to right, that is, from R1 to Rm . They should not be applied between \u2018Rj\u2019s. For any assortment of \u2018Rj\u2019s with scales of measurement among types I)\u2013IV), a single set R\u2009=\u2009{Rj| X(j)1\u2009\u00d7\u2009X(j)2\u2009\u00d7\u2009X(j)3\u2009\u00d7\u2009X(j)4\u2009\u00d7\u2009X(j)5} (\u2018\u00d7\u2019 means products among groups and fields) using a vector-like notation for the scoring of patient states can be structured where all possible assessments and/or clinical findings of the patient and treatment are included. The general form is the n-fold product; set R\u2009=\u2009{Rj| X(j)1\u2009\u00d7\u2009X(j)2\u2009\u00d7\u2009X(j)3\u2009\u00d7\u2009X(j)4\u2009\u00d7\u2009\u2026\u00d7X(j)(n-2)\u2009\u00d7\u2009X(j)(n-1)\u2009\u00d7\u2009X(j)n} .Note that, in general, there exists an identity \u2018E each with four component (\u2018n\u2009=\u20095\u2019) and arrows (only symbols) that indicate the possible changes among the \u2018Rj\u2019s, as displayed in Figure\u00a0j\u2019s, are not combined directly with each other in the sense of operations. Then, the arrows could be re-displayed according to our concepts as operators \u2018R(j\u2192k) that can be regarded as elements \u2018Rj\u2019 belonging to a set R as in Figure\u00a0j\u2019s are merely a collection of values and the arrows in Figure\u00a0j\u2019s and \u2018R(j\u2192k)\u2019s are elements of a single set R subject to axioms of an abstract algebra as indicated using composition symbol \u2018\u25ca\u2019 in Figure\u00a0j to Rk\u2019 can be traced at each session. Displayed in this way, Figure\u00a0As for the possible application to better data mining or data usage from the viewpoint of our reinterpretation, we provide a simple example that may help readers to follow an outline of the argument. Consider an example of 17 states \u201cR1\u2019 there are four outcomes \u2018R6\u2019, \u2018R9\u2019, \u2018R16\u2019, and \u2018R17\u2019 containing nodes at \u2018R2\u2019 \u2018R4\u2019 \u2018R10\u2019 \u2018R12\u2019 and \u2018R13\u2019. By making use of our previous examples \u2018R1\u2009-\u2009R3\u2019, the next simplest examples with \u2018n (component number)\u2009=\u20095\u2019 can be confirmed easily:Here, consider the scenario of Figure\u00a0Following these results, the next relations, according to the tree in Figure\u00a0The operator expressions are evaluated in Appendix F.Similarly, the next sequences are definable in principle,a to Rb (=Ra\u25caR(a\u2192b)\u2009=\u2009Rb)\u2019 and \u2018Ra to Rc (=Ra\u25caR(a\u2192c)\u2009=\u2009Rc)\u2019 as \u2018Ra[(\u25caR(a\u2192b))(\u25caR(a\u2192c))]\u2019 ; here \u2018\u2026\u2019 meaning simple juxtaposition. All paths belonging to the operational tree of Figure\u00a0In general, we denote a node divergence \u2018R6, R9, R16, and R17\u2019 and divergence point \u2018R4\u2019 a notation \u2018(=R6), (=R9), (=R16) and (=R17)\u2019 might be considered. Hence,To display for easy recognition, for example, end states like \u2018R(3\u2009\u2192\u20094)[(\u25caR(4\u2009\u2192\u20095)\u2009\u25ca\u2009R(5\u2009\u2192\u20096))(\u25caR(4\u2009\u2192\u20098)\u2009\u25ca\u2009R(8\u2009\u2192\u20099)\u2009\u25ca\u2009R(9\u2009\u2192\u200910))(\u25caR(4\u2009\u2192\u200915)\u2009\u25ca\u2009R(15\u2009\u2192\u200916))\u2009\u2026] from the left-hand side by \u2018R3\u2019. The subsequent result can be expressed in accordance with the single scheme presented in Figure\u00a0Moreover, composition with an operator as in operating on \u2018R\u2009\u2192\u20094, \u20182Rj1 = 21Rj2 \u222a 22Rj2\u2019 = [X(j)3 (/mm3) | X(j)4 (mEq/l)|X(j)5 (\u2026)], whereas at the third level, \u201822Rj2 = 221Rj3 \u222a 222Rj3\u2019 = [X(j)4 (mEq/l)|X(j)5 (\u2026)], 221Rj3 = [X(j)4 (mEq/l)], 222Rj3 = [X(j)5 (\u2026)] 1, X(j)2, X(j)3, X(j)4, X(j)5} = [X(j)1 (mod 7) | X(j)2 (/24 hrs) | X(j)3 (/mm3) | X(j)4 (mEq/l) | X(j)5 (\u2026)]. A hierarchy has additional levels as necessary to reach single units at its base \u2019 and \u2018R1 = [X(1)1| X(1)2 |\u2026| X(1)b]\u2019, \u2018{Rk, R1} =Rj [X(j)1| X(j)2 |\u2026| X(j)a | X(j)a+1| X(j)a+2 |\u2026| X(j)a+b] \u2019 . In this way, classical datasets that are classified in the Stevens scales of measurement could be mined and combined on a higher abstract structure level. To help better understand the concept, a sequence of schemes illustrating the principles of our model is presented in Figure\u00a0Additionally, we can include data mining in a more symbolic/abstract way as follows. For an arbitrary j , a hierarchical-cluster-like expression can be defined . For ins X(j)2 /2 hrs], \u20182Subject to future improvements, we envisage that this compact description is versatile to provide better data mining/data usage than from existing methods, although a final version is far from complete at this early stage.11\u2019 arises as a natural modular scale. In contrast, similar approaches might be difficult for a \u201cvisual analogue scale\u201d changes in the following manner: \u20185000 (/mm1 = 145 (mEq/l)\u2019 changes into \u2018[Na]2 = 128 (mEq/l)\u2019, because \u2018[Na]1 # [Na](1\u21922) = [Na]2\u2019, the operator for addition is obtain from \u2018[Na](1\u21922) = [Na]2 - [Na]1 = 128 - 145 = - 17 (mEq/l)\u2019. Collectively, the operator for division is \u2018[Na](1\u21922) = [Na]2/[Na]1 = 128/145 (mEq/l)\u2019.For an another example, if \u2018[Na](1\u21922) = R2 - R1R3) | 128 (mEq/l) | X(2)5 (\u2026)] - [2 (mod 7) | 21 (/24 hrs) | 5000 (/mm3) | 145 (mEq/l) | X(1)5 (\u2026)],= [5 (mod 7) | 19.5 (/24 hrs) | 18000 (/mm3) | 128 - 145 (mEq/l) | X(1\u21922)5 (\u2026)],= [5 - 2 (mod 7) | 19.5 - 21 (/24 hrs) | 18000 - 5000 (/mm3) | - 17 (mEq/l) | X(1\u21922)5 (\u2026)].= [3 (mod 7) | - 1.5 (/24 hrs) | 13000 (/mm(2\u21923) = R3 - R2R3) | 158 (mEq/l)] | X(3)5 (\u2026)] - [5 (mod 7) | 19.5 (/24 hrs) | 18000 (/mm3) | 128 (mEq/l) | X(2)5 (\u2026)],= [3 (mod 7) | 22 (/24 hrs) | 7000 (/mm3) | 158 - 128 (mEq/l) | X(2\u21923)5 (\u2026)],= [3 - 5 (mod 7) | 22 - 19.5 (/24 hrs) | 7000 - 18000 (/mm3) | 30 (mEq/l) | X(2\u21923)5 (\u2026)],= [- 2 (mod 7) | 2.5 (/24 hrs) | - 11000 (/mm3) | 30 (mEq/l) | X(2\u21923)5 (\u2026)].= [5 (mod 7) | 2.5 (/24 hrs) | - 11000 (/mm1\u25caR(1\u21922)\u25caR(2\u21923) = [2 (mod 7) | 21 (/24 hrs) | 5000 (/mm3) | 145 (mEq/l) | X(1)5 (\u2026)]\u25ca[3 (mod 7) | - 1.5 (/24 hrs) | 13000 (/mm3) | - 17 (mEq/l) | X(1\u21922)5 (\u2026)]\u25ca[5 (mod 7) | 2.5 (/24 hrs) | - 11000 (/mm3) | 30 (mEq/l) | X(2\u21923)5 (\u2026)],R3) | 145 - 17 +30 (mEq/l) | X(3)5 (\u2026)],= [2 + 3 + 5 (mod 7) | 21 - 1.5 + 2.5 (/24 hrs) | 5000 + 13000 - 11000 (/mm3) | 158 (mEq/l) | X(3)5 (\u2026)],= [10 (mod 7) | 22 (/24 hrs) | 7000 (/mm3) | 158 (mEq/l) | X(3)5 (\u2026)].= [3 (mod 7) | 22 (/24 hrs) | 7000 = R10 - R2 = [0 - 5 (mod 7) | 17 - 19.5 (/24 hrs) | 9000 - 18000 (/mm3) | 130 - 128 (mEq/l) | X(2\u219210)5 (\u2026)] = [- 5 (mod 7) | - 2.5 (/24 hrs) | - 9000 (/mm3) | 2 (mEq/l) | X(2\u219210)5 (\u2026)],R(10\u219211) = R11 - R10 = [6 - 0 (mod 7) | 20 - 17 (/24 hrs) | 20000 - 9000 (/mm3) | 149 - 130 (mEq/l) | X(10\u219211)5 (\u2026)] = [6 (mod 7) | 3 (/24 hrs) | 11000 (/mm3) | 19 (mEq/l) | X(10\u219211)5 (\u2026)],R(11\u219212) = R12 - R11 = [4 - 6 (mod 7) | 23 - 20 (/24 hrs) | 6000 - 20000 (/mm3) | 140 - 149 (mEq/l) | X(11\u219212)5 (\u2026)] = [- 2 (mod 7) | 3 (/24 hrs) | - 14000 (/mm3) | - 9 (mEq/l) | X(11\u219212)5 (\u2026)],R(10\u219213) = R13 - R10 = [1 - 0 (mod 7) | 18 - 17 (/24 hrs) | 5000 - 9000 (/mm3) | 135 - 130 (mEq/l) | X(10\u219213)5 (\u2026)] = [1 (mod 7) | 1 (/24 hrs) | - 4000 (/mm3) | 5 (mEq/l) | X(10\u219213)5 (\u2026)],R(13\u219217) = R17 - R13 = [2 - 1 (mod 7) | 23.5 - 18 (/24 hrs) | 3000 - 5000 (/mm3) | 150 - 135 (mEq/l) | X(13\u219217)5 (\u2026)] = [1 (mod 7) | 5.5 (/24 hrs) | - 2000 (/mm3) | 15 (mEq/l) | X(13\u219217)5 (\u2026)].RThe authors declare that they have no competing interests.JS conceived the main concept of this article and wrote the manuscript. SM revised the manuscript. JI gave advice on the potential validity from the viewpoint of clinical research and treatment. Additionally, all authors read and approved the final manuscript."} +{"text": "The aromatic substituents on the sulfonate group are oriented gauche to one another with a C\u2014O\u2014S\u2014C torsion angle of \u221262.0\u2005(3)\u00b0. The supra\u00admolecular features that contribute to the crystal lattice are offset \u03c0-\u03c0 and multiple C\u2014H\u22efO inter\u00adactions.The title compound, C 13H10N2O7S, was synthesized via a nucleophilic substitution reaction between 2,4-di\u00adnitro\u00adphenol and p-toluene\u00adsulfonyl chloride. This crystal structure is a polymorph of CSD entry WUVYUH . The aromatic substituents on the sulfonate group are oriented gauche to one another with a C\u2014O\u2014S\u2014C torsion angle of \u221262.0\u2005(3)\u00b0. The supra\u00admolecular features that contribute to the crystal stability are offset \u03c0\u2013\u03c0 [centroid\u2013centroid distance = 3.729\u2005(2)\u2005\u00c5] and multiple C\u2014H\u22efO inter\u00adactions.The title compound, C Analogous to the carbonyl group, nucleophilic substitution reactions of sulfonyl derivatives have also been reported reactions similar to those reported by others . The bond angle between the S=O groups (O1\u2014S1\u2014O2) is 121.20\u2005(17)\u00b0, while that of the aromatic substituents (O3\u2014S1\u2014C5) is 103.27\u2005(15)\u00b0. The two aromatic rings are in a gauche orientation about the O3\u2014S1 bond with a torsion angle (C8\u2014O3\u2014S1\u2014C5) of \u221262.0\u2005(3)\u00b0.The central sulfur atom (S1) is tetra\u00adhedral with S=O bond lengths of 1.415\u2005(3) and 1.414\u2005(3)\u2005\u00c5, and an S\u2014O bond length of 1.634\u2005(3)\u2005\u00c5 Fig.\u00a02a. The bet al., 2003ab). While the bond lengths of the two polymorphs agree within 0.01\u2005\u00c5 of each other, there are some differences between bond angles. The aromatic rings in WUVYUH are in an anti orientation along the S\u2014O bond, with a torsion angle of 141.02\u2005(9)\u00b0. The bond angle between the S=O groups (O1\u2014S1\u2014O2) is 119.80\u2005(6)\u00b0, while that of the aromatic substituents (O3\u2014S1\u2014C5) is 98.17\u2005(5)\u00b0.For comparison, the polymorph WUVYUH = 3.379\u2005(4)\u2005\u00c5, and O7(nitro)\u22efS1v(sulfonic ester) = 3.877\u2005(3)\u2005\u00c5. The relatively short inter\u00admolecular distance between N2 and O17 suggests the presence of favorable N\u22efO inter\u00adactions in the crystal is in proximity to the sulfonic ester of the C2\u2013C7et al., 2008ortho-substit\u00aduent is a nitro group and the para-position bears a second tosyl\u00adate group. The remaining entries have various electron-rich groups in the ortho-position including meth\u00adoxy .The CSD contains three additional structures where the position p-toluene\u00adsulfonyl chloride (5\u2005mmol) and pyridine (3\u2005mmol) in 10\u2005mL of di\u00adchloro\u00admethane for 30 minutes at room temperature. The reaction was heated to 353\u2005K for 30 minutes in a microwave reactor, then cooled to room temperature and stirred overnight in a fume hood. The reaction mixture was transferred to a scintillation vial where the pale yellow product crystallized upon standing after several days and was filtered from the mother liquor (m.p. 393.4\u2013394.7\u2005K).The title compound was prepared by stirring 2,4-di\u00adnitro\u00adphenol (5\u2005mmol), Uiso(H) = 1.2Ueq(C) for CH groups and Uiso(H) = 1.5 Ueq(C) for methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015015650/pk2562sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015015650/pk2562Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015015650/pk2562Isup3.cmlSupporting information file. DOI: 1419864CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-tosyl\u00adacryl\u00adamide compounds, (I) and (II), the conformation about the C=C bond is E. In (I), the furan, phenyl and 4-methyl\u00adbenzene rings are inclined to the acryl\u00adamide mean plane [\u2013NH\u2014C(= O)\u2014C=C\u2013] by 26.47\u2005(11), 69.01\u2005(8) and 82.49\u2005(9)\u00b0, respectively. In (II), the phenyl, and 3-methyl and 4-methyl\u00adbenzene rings are inclined to the acryl\u00adamide mean plane by 11.61\u2005(10), 78.44\u2005(10) and 78.24\u2005(10)\u00b0, respectively. In the crystals of both compounds, mol\u00adecules are linked by pairs of N\u2014H\u22efO hydrogen bonds, forming inversion dimers with In the title N-tosyl\u00adacryl\u00adamide compounds, C20H17NO4S, (I), and C23H21NO3S, (II), the conformation about the C=C bond is E. The acryl\u00adamide groups, [\u2013NH\u2014C(=O)\u2014C=C\u2013], are almost planar, with the N\u2014C\u2014C=C torsion angle being \u2212170.18\u2005(14)\u00b0 in (I) and \u2212168.01\u2005(17)\u00b0 in (II). In (I), the furan, phenyl and 4-methyl\u00adbenzene rings are inclined to the acryl\u00adamide mean plane by 26.47\u2005(11), 69.01\u2005(8) and 82.49\u2005(9)\u00b0, respectively. In (II), the phenyl, 3-methyl\u00adbenzene and 4-methyl\u00adbenzene rings are inclined to the acryl\u00adamide mean plane by 11.61\u2005(10), 78.44\u2005(10) and 78.24\u2005(10)\u00b0, respectively. There is an intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction present in compound (II). In the crystals of both compounds, mol\u00adecules are linked by pairs of N\u2014H\u22efO hydrogen bonds, forming inversion dimers with an R22(8) ring motif. In (I), the dimers are reinforced by C\u2014H\u22efO hydrogen bonds and linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming chains along [011]. In the crystal of (II), the dimers are linked via C\u2014H\u22efO hydrogen bonds, forming chains along [100]. The chains are further linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming layers parallel to (010).In the title There is also a C\u2014H\u22ef\u03c0 inter\u00adaction present, linking the chains to form layers lying parallel to (010).In the crystal of both compounds, mol\u00adecules are linked by pairs of N\u2014H\u22efO hydrogen bonds Tables 1 and 2 \u25b8,et al., 2016N-(phenyl\u00adsulfon\u00adyl)acryl\u00adamide yielded five hits. Four of these compounds involve the 4-methyl\u00adbenzene\u00adsulfonyl group and one compound involves a phenyl\u00adsulfonyl group. This later compound, 2-(4-chloro\u00adphen\u00adyl)-3-(2-fur\u00adyl)-N-(phenyl\u00adsulfon\u00adyl)acryl\u00adamide : 4-methyl\u00adbenzene\u00adsulfonyl azide (4.5\u2005mmol), CuI , Et4NI , ethynyl\u00adbenzene (4.5\u2005mmol), and furan-2-carbaldehyde (3\u2005mmol) were suspended in CH2Cl2 (5\u2005ml) in a 10\u2005mL Schlenk tube under nitro\u00adgen at rt. LiOH was then added, and the resulting solution was stirred at this temperature. Upon full consumption of furan-2-carbaldehyde, the reaction was quenched by saturated aqueous NH4Cl (5\u2005ml) and extracted with CH2Cl2 (10\u2005ml \u00d7 3). The combined organic layers were dried over anhydrous Na2SO4 and concentrated in vacuo. The crude residue was purified by column chroma\u00adtography on silica gel (n-hexa\u00adne/EtOAc 5:1 v/v) to afford compound (I)n-hexa\u00adne/EtOAc and after slow evaporation over several days, colourless crystals suitable for analysis by X-ray diffraction were formed.Compound (II): 4-methyl\u00adbenzene\u00adsulfonyl azide (4.5\u2005mmol), CuI , Et4NI , 1-eth\u00adyn\u00adyl-3-methyl\u00adbenzene (4.5\u2005mmol), and benzaldehyde (3\u2005mmol) were suspended in CH2Cl2 (5\u2005ml) in a 10\u2005mL Schlenk tube under nitro\u00adgen at rt. LiOH was then added, and the resulting solution was stirred at this temperature. Upon full consumption of benzaldehyde, the reaction was quenched by saturated aqueous NH4Cl (5\u2005ml) and extracted with CH2Cl2 (3 \u00d7 10\u2005ml). The combined organic layers were dried over anhydrous Na2SO4 and concentrated in vacuo. The crude residue was purified by column chroma\u00adtography on silica gel (n-hexa\u00adne/EtOAc 5:1 v/v) to afford compound (II)n-hexa\u00adne/EtOAc and after slow evaporation over several days, colourless block-like crystals were obtained.Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016007611/su5296sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: Click here for additional data file.10.1107/S2056989016007611/su5296Isup2.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016007611/su5296IIsup3.cmlSupporting information file. DOI: 1478730, 1478729CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, supra\u00admolecular chains are formed 13H10N2O2 , features an almost planar (r.m.s. deviation = 0.0095\u2005\u00c5) central C3O2 core consolidated by an intra\u00admolecular hy\u00addroxy-O\u2014H\u22efO(carbon\u00adyl) hydrogen bond. Twists are evident in the mol\u00adecule, as seen in the dihedral angles between the central core and the 2- and pyridin-3-yl rings of 8.91\u2005(7) and 15.88\u2005(6)\u00b0, respectively. The conformation about the C=C bond [1.3931\u2005(17)\u2005\u00c5] is Z, and the N atoms lie to the same side of the mol\u00adecule. In the mol\u00adecular packing, supra\u00admolecular chains along the a axis are mediated by \u03c0(pyridin-2-yl)\u2013\u03c0(pyridin-3-yl) inter\u00adactions [inter-centroid distance = 3.7662\u2005(9)\u2005\u00c5]. The observation that chains pack with no directional inter\u00adactions between them is consistent with the calculated electrostatic potential, which indicates that repulsive inter\u00adactions dominate.The title compound, C III and GaIII octa\u00adhedral building blocks for network structures linked by AgI ions -2,4-penta\u00adnedione as well as the mixed-MOF with AgNO3 in a two-dimensional honeycomb structure while at higher AgI concentrations, a one-dimensional ladder motif was formed versus hy\u00addroxy-O atoms is not readily confirmed by a great disparity in the C1\u2014O1 [1.2871\u2005(14)\u2005\u00c5] and C3\u2014O2 [1.3041\u2005(14)\u2005\u00c5] bond lengths. The assignment was based on an unrestrained refinement of the H1O atom which resulted in a O2\u2014H1O bond length of 1.090\u2005(18)\u2005\u00c5. More certainty is associated with the assignment of the nitro\u00adgen atoms in the pyridyl rings. Thus, the short C5\u2014N1 and C4\u2014N1 [1.3325\u2005(17) and 1.3484\u2005(15)\u2005\u00c5] and C10\u2014N2 and C11\u2014N2 [1.3371\u2005(17) and 1.3397\u2005(18)\u2005\u00c5] bond lengths cf. the C\u2014C bonds in the rings confirm their assignment. The central C3O2 residual in (I)syn arrangement of the oxygen atoms enables the formation of an intra\u00admolecular hy\u00addroxy-O\u2014H\u22efO(carbon\u00adyl) hydrogen bond, Table\u00a01Z, and, to a first approximation, the N1 and N2 atoms lie to the same side of the mol\u00adecule.In (I)Cg(N1-pyridin\u00adyl)\u22efCg(N2-pyridin\u00adyl) = 3.7662\u2005(9)\u2005\u00c5, angle of inclination = 7.45\u2005(6)\u00b0 for symmetry operation 1\u00a0+\u00a0x, y, z]. The result is the formation of a linear supra\u00admolecular chain, Fig.\u00a02a. The chains pack with no directional inter\u00adactions between them in accord with the distance criteria in PLATON , is viewed as bright-red spots near these atoms on the Hirshfeld surface mapped over dnorm, Fig.\u00a04From the Hirshfeld surface mapped over electrostatic potential, Fig.\u00a03gs Fig.\u00a02a, is via, and those delin\u00adeated into H\u22efH, C\u22efC, O\u22efH/H\u22efO, C\u22efH/H\u22efC and N\u22efH/H\u22efN contacts are illustrated in Fig.\u00a05b\u2013f, respectively; their relative contributions to the surface are qu\u00adanti\u00adfied in Table\u00a02et al., 2007b, with a single peak at less than the van der Waals separation corresponding to a short H13\u22efH13 contact of 2.33\u2005\u00c5 . The short inter\u00adatomic C5\u22efC10 contact and \u03c0\u2013\u03c0 stacking inter\u00adactions appear as an arrow-like distribution of points with the tip at de + di \u223c 3.3\u2005\u00c5 . The presence of \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridyl rings is also apparent from the appearance of red and blue triangle pairs on the Hirshfeld surface mapped with shape-index property identified with arrows in the image of Fig.\u00a06The overall 2D fingerprint plot, Fig.\u00a05\u2005\u00c5 Fig.\u00a05c. The pd\u2013f. However, the points are distributed at distances greater than their respective van der Waals separations. This is consistent with the repulsion between the atoms having electrostatic negative potential dominating the mol\u00adecular packing, hence the lack of specific inter\u00admolecular inter\u00adactions between supra\u00admolecular chains.The two-dimensional fingerprint plots delineated into O\u22efH/H\u22efO, C\u22efH/H\u22efC and N\u22efH/H\u22efN inter\u00adactions exhibit their usual characteristic features in their respective plots; Fig.\u00a04et al., 2016et al. solution afforded colourless crystals. Yield: 4.03\u2005g (70.7%). M.p: 377\u2013378\u2005K. IR (KBr pellet) \u03bdmax/cm\u22121: 3121 (m), 3053 (m), 2922 (m), 2853 (m), 1611 (s), 1595 (s), 1539 (m), 1458 (m), 1418 (m), 1221 (m), 1188 (m), 1146 (m), 1115 (m), 1067 (m), 1018 (m), 989 (m), 926 (m), 775 (s), 739 (m), 679 (s), 611 (m). Analysis calculated for C13H10N2O2: C, 69.03; H, 4.42; N, 12.19. Found: C, 68.73; H, 4.54; N, 12.16. MS: m/z 226. 1H NMR \u03b4 9.22 , 8.82 , 8.44 , 8.17 , 8.09 , 7.70 , 7.63 .2-Acetyl\u00adpyridine was added to a suspension of NaH in anhydrous THF (10\u2005ml) at room temperature with stirring. Ethyl nicotinate in anhydrous THF (10ml) was added dropwise to the mixture over 3\u2005min. The yellow mixture was refluxed under a nitro\u00adgen atmosphere for 1.3\u2005h and then quenched with ice\u2013water (50\u2005ml). Glacial acetic acid was added to adjust the pH to 6\u20137. The resulting yellow precipitate was collected by filtration, washed with cold water and dried under vacuum. Recrystallization from di\u00adchloro\u00admethane\u2013hexane (1:1 Uiso(H) set to 1.2Ueq(C). The hy\u00addroxy-H atom was located in a difference map and refined with O\u2014H = 0.82\u00b10.01\u00c5, and with Uiso(H) set to 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S205698901600832X/hb7587sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901600832X/hb7587Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901600832X/hb7587Isup3.cmlSupporting information file. DOI: 1481225CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A and B) in the asymmetric unit. In the crystal, individual mol\u00adecules are linked by pairs of O\u2014H\u22efO hydrogen bonds, forming A\u2013A and B\u2013B inversion dimers with The title compound, crystallized with two conformationally similar mol\u00adecules in the asymmetric unit. The two mol\u00adecules have the same conformation; the mol\u00adecular overlap gives weighted and unit-weight r.m.s. fits of 0.047 and 0.043\u2005\u00c5, respectively. The conformation of the N-(hydroxeth\u00adyl)formamide chains are very similar, as indicated by the C\u2014N(H)\u2014C=O and C\u2014N(H)\u2014C\u2014O(H) torsion angles, which are, respectively, \u22121.8\u2005(3) and \u221291.5\u2005(2)\u00b0 for mol\u00adecule A, and \u22122.1\u2005(3) and \u221295.7\u2005(2)\u00b0 for mol\u00adecule B. In the crystal, individual mol\u00adecules are linked by pairs of O\u2014H\u22efO hydrogen bonds, forming A\u2013A and B\u2013B inversion dimers with R22(12) ring motifs. The dimers are linked via N\u2014H\u22efO hydrogen bonds, forming alternating layers of A and B mol\u00adecules parallel to the bc plane. Within the layers of B mol\u00adecules, there are weak C\u2014H\u22efCl hydrogen bonds present.The title compound, C This has led to the use of formamides as key inter\u00admediates in numerous organic synthetic endeavours (Kobayashi A and B) in the asymmetric unit. The arbitrarily chosen chirality of atoms C2 in mol\u00adecule A and C5 in mol\u00adecule B is the same. The backbones of the two mol\u00adecules have almost identical conformations with weighted and unit-weight r.m.s. overlay fits of 0.047 and 0.043\u2005\u00c5, respectively, for the six atoms in each mol\u00adecule formamide, yielded 25 hits. The majority concern metal complexes of the ligand N-(hy\u00addroxy\u00admeth\u00adyl)nicotinamide. Only one compound, N,N\u2032-diformamide formamide chain as indicated by the C\u2014N(H)\u2014C\u2014O(H) and C\u2014N(H)\u2014C=O torsion angles: 1.6\u2005(2) and \u221299.09\u2005(14)\u00b0 for the above mentioned compound compared to \u22121.8\u2005(3) and \u221291.5\u2005(2)\u00b0 for mol\u00adecule A and \u22122.1\u2005(3) and \u221295.7\u2005(2)\u00b0 for mol\u00adecule B of the title compound formamide was dissolved in hot ethanol, followed by treatment with charcoal. The filtered solution was left to crystallize by slow evaporation, forming colourless block-like crystals (m.p. 393\u2005K).The title compound can be synthesized following a literature procedure I. DOI: 10.1107/S2056989015020459/hg5462Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015020459/hg5462Isup3.cmlSupporting information file. DOI: 1009715CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two aromatic rings present in the mol\u00adecule are almost coplanar, forming a dihedral angle of 1.4\u2005(2)\u00b0. The five-membered ring involving the metal atom is puckered, with an amplitude Q = 0.358\u2005(2)\u2005\u00c5 and \u03d5 = 204.1\u2005(6)\u00b0. In the crystal, pairs of inversion-related mol\u00adecules are linked by O\u2014H\u22efN hydrogen bonds. An O\u2014H\u22efO hydrogen bond connects the water ligand to the di\u00admethyl\u00adformamide solvent mol\u00adecule. The crystal packing also features \u03c0\u2013\u03c0 [centroid\u2013centroid distance of 3.688\u2005(2)\u2005\u00c5] and C\u2014H\u22efO inter\u00adactions.The title compound, [Mo(C The complex adopts a distorted octa\u00adhedral geometry around the Mo atom \u2005\u00c5] is close to the reported C\u2014O single bond length and \u22121.4\u2005(4)\u00b0, respectively]. This configuration is similar to that of the metal-free ligand \u00c5 and \u03d5 = 204.1\u2005(6)\u00b0.The ligand adopts D\u22efA distances 2.891\u2005(4) and 2.701\u2005(4)\u2005\u00c5 respectively, and a non-classical C\u2014H\u22efO inter\u00adaction with a D\u22efA distance of 3.421\u2005(5)\u2005\u00c5. These inter\u00adactions connect pairs of mol\u00adecules along with the solvent di\u00admethyl\u00adformamide. The complex mol\u00adecule is stacked along the b axis through two different types of O\u2014H\u22ef\u03c0 inter\u00adaction with a centroid-centroid distance of 3.688\u2005(2)\u2005\u00c5 for 3\u2005h. The brown precipitate obtained was filtered, washed with methanol, dried and recrystallized from di\u00admethyl\u00adformamide . FT\u2013IR \u03bdmax: 3400, 3194, 1657, 1546, 1345, 937, 810.The complex was synthesized by refluxing a methano\u00adlic solution of benzoyl hydrazone and MoClUiso(H) = 1.2Ueq(carrier) or 1.5Ueq(methyl C). The O\u2014H distances were restrained with 1,2 and 1,3 distance restraints of 0.86\u2005(1) and 1.36\u2005(2)\u2005\u00c5. Reflections (0 0 2), (1 0 1) and I. DOI: 10.1107/S2056989015009639/pk2550Isup2.hklStructure factors: contains datablock(s) I. DOI: 1401828CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Weak C\u2014H\u22efCl and C\u2014H\u22efS hydrogen-bonding inter\u00adactions consolidate the packing of the structure.The title cobalt(II) complex consists of a CoCl 3CS)2C=NC\u00a0 N}2CoCl2], consists of a CoII atom coordinated in a distorted tetra\u00adhedral manner by two Cl\u2212 ligands and the terminal N atoms of two dimethyl N-cyano\u00addithio\u00adimino\u00adcarbonate ligands. The two organic ligands are almost coplanar, with a dihedral angle of 5.99\u2005(6)\u00b0 between their least-squares planes. The crystal packing features pairs of inversion-related complexes that are held together through C\u2014H\u22efCl and C\u2014H\u22efS inter\u00adactions and \u03c0\u2013\u03c0 stacking [centroid-to-centroid distance = 3.515 (su?) \u00c5]. Additional C\u2014H\u22efCl and C\u2014H\u22efS inter\u00adactions, as well as Cl\u22efS contacts < 3.6\u2005\u00c5, consolidate the crystal packing.The structure of the mononuclear title complex, [{(H N-cyano\u00addithio\u00adimino\u00adcarbonate with its two N and two S atoms has four possible coordination sites and hence should present a high coordination ability. The behaviour of N and S atoms according to Pearson\u2019s concept as hard and soft donors, respectively, may allow coordination to both hard and soft Lewis acids. Despite this coordination property, the ligand has scarcely been studied. Only one crystalline compound with dimethyl N-cyano\u00addithio\u00adimino\u00adcarbonate as a ligand has been reported previously chloride hexa\u00adhydrate and dimethyl N-cyano\u00addithio\u00adimino\u00adcarbonate which has yielded the title complex, [{(H3CS)2C=NC\u00a0 N}2CoCl2].Dimethyl II atom coordinated in a distorted tetra\u00adhedral manner by two Cl\u2212 ligands and the cyanide N atoms of two dimethyl N-cyano\u00addithio\u00adimino\u00adcarbonate ligands \u00b0]. The CoII atom lies 0.437\u2005(2) and 0.557\u2005(2)\u2005\u00c5 from the mean planes of the two ligands.The structure of the title complex consists of a Cods Fig.\u00a01. Co\u2014Cl ads Fig.\u00a01. The Cl\u2014t Table\u00a01. Despite3C\u2014S groups of the adjacent mol\u00adecule, presumably reducing steric inter\u00adactions. Apart from C\u2014H\u22efCl and C\u2014H\u22efS inter\u00adactions (Table\u00a02(su?) \u00c5 prevails within a pair of complex mol\u00adecules. In the crystal, these pairs are arranged parallel to (110) Fig.\u00a02. Additio0) Fig.\u00a02.2\u00b76H2O in aceto\u00adnitrile (30\u2005ml) and dimethyl N-cyano\u00addithio\u00adimino\u00adcarbonate in aceto\u00adnitrile (20\u2005ml) at room temperature. The resulting blue solution was stirred for about 2\u2005h. Blue crystals suitable for single-crystal X-ray diffraction were obtained after six days of slow solvent evaporation at room temperature (300\u2005K).All chemicals were purchased from Aldrich (Germany) and were used as received. The title compound was prepared by mixing of CoCl\u03bd(C\u00a0N) 2224\u2005cm\u22121, \u03bd(C=N) 1458\u2005cm\u22121, \u03bd(CS2) + rocking CH3 1024 and 962\u2005cm\u22121. Melting point 398\u2005K. Elemental analyses of C8H12Cl2CoN4S4: calculated (found): C 22.75 (21.91), H 2.86 (3.43), N 13.27 (12.63), S 30.37 (29.40).Infra-red bands: Uiso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989015023439/wm5249sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015023439/wm5249Isup2.hklStructure factors: contains datablock(s) I. DOI: 1440755CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II complex, the MnII atom is coordinated by two 4-nitro\u00adbenzoate (NB) anions, two nicotinamide (NA) ligands and two water mol\u00adecules; the NB and NA ligands act as monodentate ligands. The resulting MnN2O4 coordination polyhedron is a distorted octa\u00adhedron.In the title centrosymmetric Mn 7H4NO4)2(C6H6N2O)2(H2O)2], contains one MnII atom, one 4-nitro\u00adbenzoate (NB) anion, one nicotinamide (NA) ligand and one water mol\u00adecule; NA and NB each act as a monodentate ligand. The MnII atom, lying on an inversion centre, is coordinated by four O atoms and two pyridine N atoms in a distorted octa\u00adhedral geometry. The water mol\u00adecules are hydrogen bonded to the carboxyl\u00adate O atoms. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 24.4\u2005(3)\u00b0, while the benzene and pyridine rings are oriented at a dihedral angle of 86.63\u2005(11)\u00b0. In the crystal, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules, forming a layer parallel to the ab plane. The layers are further linked via weak C\u2014H\u22efO hydrogen bonds, a \u03c0\u2013\u03c0 stacking inter\u00adaction [centroid\u2013centroid distance = 3.868\u2005(2)\u2005\u00c5] and a weak C\u2014H\u22ef\u03c0 inter\u00adaction, resulting in a three-dimensional network.The asymmetric unit of the title compound, [Mn(C A deficiency of this vitamin leads to loss of copper from the body, known as pellagra disease. The NA ring is the reactive part of nicotinamide adenine dinucleotide (NAD) and its phosphate (NADP), which are the major electron carriers in many biological oxidation\u2013reduction reactions \u00admanganese \u00adbis\u00ad(4-nitro\u00adbenzoato)manganese of the two symmetry-related monodentate NB anions and the two symmetry-related water O atoms (O6 and O6iii) around the MnII atom form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination sphere is completed by the two pyridine N atoms (N2 and N2iii) of the two symmetry-related monodentate NA ligands in the axial positions in the carboxyl\u00adate group indicates delocalized bonds rather than localized single and double bonds. The Mn\u2014O bond lengths [2.156\u2005(2) and 2.115\u2005(2)\u2005\u00c5] and the Mn\u2014N bond length [2.134\u2005(3)\u2005\u00c5] are close to the standard values. Atom Mn1 lies 0.4172\u2005(1)\u2005\u00c5 above the O1/O2/C1 plane of the carboxyl\u00adate group. The O\u2014Mn\u2014O and O\u2014Mn\u2014N bond angles deviate slightly from the ideal value of 90\u00b0. The dihedral angle between the carboxyl\u00adate group (O1/O2/C1) and the adjacent benzene (C2\u2013C7) ring is 24.4\u2005(3)\u00b0, while the benzene ring and the pyridine (N2/C8\u2013C12) ring are oriented at a dihedral angle of 86.63\u2005(11)\u00b0.na\u22efOna (na = nicotinamide), N\u2014Hna\u22efOc (c = carboxyl\u00adate group) and O\u2014Hw\u22efOna (w = water) hydrogen bonds \u2005\u00c5; symmetry code: (ix) 1\u00a0\u2212\u00a0x, \u2212y, 1\u00a0\u2212\u00a0z, where Cg1 is the centroid of the C2\u2013C7 ring].In the crystal, inter\u00admolecular N\u2014Hs Table\u00a01 link thene Fig.\u00a02. In the n Table\u00a01 and a \u03c0\u20134\u00b7H2O in H2O (25\u2005ml) and nicotinamide in H2O (25\u2005ml) with sodium 4-nitro\u00adbenzoate in H2O (150\u2005ml). The mixture was filtered and set aside to crystallize at ambient temperature for one week, giving colourless single crystals.The title compound was prepared by the reaction of MnSOA and H3B of the NH2 group were located in a difference Fourier map, and their coordinates were refined with distance restraints of O\u2014H = 0.85\u2005(2)\u2005\u00c5 and N\u2014H = 0.86\u2005(2)\u2005\u00c5, and with Uiso(H) = 1.5Ueq. The C-bound H atoms were positioned geometrically with C\u2014H = 0.93\u2005\u00c5 and were constrained to ride on their parent atoms with Uiso(H) = 1.2Ueq(C).The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0210.1107/S2056989016005612/is5449sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016005612/is5449Isup2.hklStructure factors: contains datablock(s) I. DOI: 1472331CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III atom in the title mol\u00adecule adopts a distorted octa\u00adhedral coordination sphere, being C,N-chelated by two 2-(pyridin-2-yl)phenyl ligands and N,O-chelated by one ancillary 2-[(phenyl\u00adimino)\u00admeth\u00adyl]phenolate ligand. The crystal packing is stabilized by inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 inter\u00adactions.The Ir 11H8N)2(C13H10NO)]\u00b7CH2Cl2, the IrIII ion is six-coordinated by two C,N-bidentate 2-(pyridin-2-yl)phenyl ligands and one N,O-bidentate 2-[(phenyl\u00adimino)\u00admeth\u00adyl]phenolate anion, giving rise to a distorted octa\u00adhedral environment. The C,N-bidentate ligands, in which the C and N atoms are statistically disordered over two sites and therefore both pairs of C and N atoms are trans and cis relative to each other, are almost perpendicular to each other [the dihedral angle between the least-square planes is 87.00\u2005(4)\u00b0]. An intra\u00admolecular C\u2014H\u22efO hydrogen bond, as well as inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 inter\u00adactions, contribute to the stabilization of the mol\u00adecular and crystal structure.In the title compound, [Ir(C III complexes are of great inter\u00adest due to their excellent phospho\u00adrescent properties and electroluminescence applications. In particular, heteroleptic IrIII complexes with imine-based ancillary ligands exhibit aggregation-induced phospho\u00adrescent emission (AIPE), resulting in enhanced phospho\u00adrescence phenomena in the solid state 2(C13H10NO)]\u00b7CH2Cl2, a heteroleptic IrIII complex with an ancillary salicyl\u00adimine ligand.Cyclo\u00admetallated IrIII ion, two 2-(pyridin-2-yl)phenyl ligands, and one 2-[(phenyl\u00adimino)\u00admeth\u00adyl]phenolate anion. The IrIII ion adopts a distorted octa\u00adhedral coordination geometry, being N,O-chelated by the 2-[(phenyl\u00adimino)\u00admeth\u00adyl]phenolate ligand and C,N-chelated by two 2-(pyridin-2-yl)phenyl ligands, in which the C and N atoms are equally disordered over two sites and therefore both pairs of C and N atoms are trans and cis relative to each other. The equatorial plane is formed by N1/O1/N2/C12 atoms, the mean deviation from the least-squares plane being 0.002\u2005\u00c5. The IrIII ion is displaced by 0.0481\u2005(9)\u2005\u00c5 from the equatorial plane towards the axial imino N3 atom. The C,N-bidentate ligands are nearly perpendicular to each other, with a dihedral angle between the least-squares planes of 87.00\u2005(4)\u00b0. Within the C,N-bidentate ligands, the dihedral angles between the aromatic rings are 3.70\u2005(10) (between rings C1\u2013C6 and N1/C7\u2013C11) and 7.67\u2005(16)\u00b0 (between rings C12\u2013C17 and N2/C18\u2013C22). As shown in Table\u00a01III compounds, e.g. {(E)-2-[\u00admeth\u00adyl]phenolato-\u03ba2N,O}bis\u00adiridium(III) -2-[\u00admeth\u00adyl]phenolato-\u03ba2N,O}bis\u00adiridium(III) -2-[(phenyl\u00adimino)\u00admeth\u00adyl]phenolato-\u03ba2N,O}bis\u00adiridium(III) \u2005\u00c5 and Cg3\u22efCg4 = 3.8873\u2005(17)\u2005\u00c5; Cg1, Cg3 and Cg4 are the centroids of the N1/C7\u2013C11, C12\u2013C17 and C30\u2013C35 rings, respectively; symmetry code: (ii) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01] contribute to the stabilization of the crystal structure = 1.2Ueq of the parent atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016008100/wm5290sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989016008100/wm5290Isup2.hklStructure factors: contains datablock(s) I. DOI: 1480710CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II cation while a water mol\u00adecule further coordinates to the CuII cation to complete the elongated distorted octa\u00adhedral coordination geometry.In a hydrated copper(II) complex, 2-amino-7-methyl-4-oxidopteridine-6-carboxyl\u00adate and 1,10-phenanthroline ligands chelate the Cu 8H5N5O3)(C12H8N2)(H2O)]\u00b73H2O, the CuII cation is O,N,O\u2032-chelated by the 2-amino-7-methyl-4-oxidopteridine-6-carboxyl\u00adate anion and N,N\u2032-chelated by the 1,10-phenanthroline (phen) ligand. A water mol\u00adecule further coordinates to the CuII cation to complete the elongated distorted octa\u00adhedral coordination geometry. In the mol\u00adecule, the pteridine ring system is essentially planar [maximum deviation = 0.055\u2005(4)\u2005\u00c5], and its mean plane is nearly perpendicular to the phen ring system [dihedral angle = 85.97\u2005(3)\u00b0]. In the crystal, N\u2014H\u22efO, O\u2014H\u22efN and O\u2014H\u22ef\u00b7O hydrogen bonds, as well as weak C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, link the complex mol\u00adecules and lattice water mol\u00adecules into a three-dimensional supra\u00admolecular architecture. Extensive \u03c0\u2013\u03c0 stacking between nearly parallel aromatic rings of adjacent mol\u00adecules are also observed, the centroid-to-centroid distances being 3.352\u2005(2), 3.546\u2005(3), 3.706\u2005(3) and 3.744\u2005(3)\u2005\u00c5.In the title compound, [Cu(C Location of the pyrazine ring N atom (N6) in the equatorial plane is in agreement with earlier observations on related copper and cobalt complexes \u2005\u00c5] may be elucidated in the light of Joule\u2019s hypothesis complexes chelated by the pterin-6-carboxyl\u00adate anion have been reported by Kohzuma al. 1989 and Funa al. 1999. In both8H7N5O3\u00b71.5H2O) was obtained by a published procedure containing Cu(NO3)2\u00b73H2O , 1,10-phenanthroline monohydrate and pterin dissolved in NaOH for 60\u2005h at 310\u2013312\u2005K under subdued light; additional NaOH solution was added for adjusting the initial pH at 10.5. Within a short while the initial bright-green solution turned hazy blue due to the presence of a fine white precipitate which gradually disappeared substanti\u00adally. The final blue solution was slightly hazy. Upon storage under aerobic conditions for one week the clear blue solution yielded green crystals, suitable for X-ray structure determination. Analysis calculated for C20H21CuN7O7: C 44.90, H 3.93, N 18.33%; found: C 44.38, H 4.06, N 17.65%. ESIMS data: the mol\u00adecular ion peak [M + 2H]+ appeared at 536.4 (relative abundance = 41.2%); the [M \u2212 4H2O \u2212 3H]+ peak was observed at 459.2 (relative abundance = 100%), indicating stability of the desolvated ternary species arising from the title complex.Method B. Using NaBH4 reduction in equimolar proportion of the original complex (obtained by Method A) and subsequent aerial reoxidation of the reduced complex to the present crystals merits attention due to the involvement of intricate redox chemistry. The NaBH4 treatment to be Na2[Cu2I(L\u2032)2(phen)(H2O)4]\u00b72H2O, where L\u2032 is the 7,8-di\u00adhydro form of the present pterin ligand anion (C8H5N5O3) .Method B, represent better accuracy [R = 0.057 and wR(F2)= 0.135] as compared to the other one [R = 0.113 and wR(F2) = 0.279].Although the title compound could be obtained by two alternative methods, the present structural data obtained using the crystals from E\u00b0\u2032 value of \u22120.68\u2005V; now using an E\u00b0\u2032 value of \u22120.80\u2005V for NaBH4 in neutral medium for the FeIII\u2013tetra\u00adhydro\u00adbiopterin reduction in phenyl\u00adalanine hy\u00addroxy\u00adlase ; using an E\u00b0\u2032 value of 0.70\u2005V for the O2/H2O2 couple, an Ecell value of 1.37\u2005V is obtained, indicating facile aerial oxidation. Now using an E\u00b0\u2032 value of 0.19\u2005V for the chelated pterin ligand = 1.2\u20131.5Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S1600536814022302/xu5822sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S1600536814022302/xu5822Isup2.hklStructure factors: contains datablock(s) I. DOI: 1028413CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The highlighted ruthenium complex forms discrete dimers by \u03c0\u2013\u03c0 stacking inter\u00adactions and hydrogen bonds. This combination of inter\u00adactions results in an unusual nearly face-to-face \u03c0\u2013\u03c0 stacking mode. 14H16N4)(C16H8N4)](PF6)2\u00b71.422CH3CN, discrete dimers of complex cations, [Ru(L\u2013N4H2)tape]2+ are formed {L\u2013N4H2 = 2,11-di\u00adaza\u00ad[3.3]pyridino\u00adphane; tape = 1,6,7,12-tetra\u00adaza\u00adperylene}, held together by \u03c0\u2013\u03c0 stacking inter\u00adactions via the tape ligand moieties with a centroid\u2013centroid distance of 3.49\u2005(2)\u2005\u00c5, assisted by hydrogen bonds between the non-coordinating tape ligand \u03b1,\u03b1\u2032-di\u00adimine unit and the amine proton of a 2,11-di\u00adaza\u00ad[3.3]-pyridino\u00adphane ligand of the opposite complex cation. The combination of these inter\u00adactions leads to an unusual nearly face-to-face \u03c0\u2013\u03c0 stacking mode. Additional weak C\u2014H\u22efN, C\u2014H\u22efF, N\u2014H\u22efF and P\u2014F\u22ef\u03c0-ring (with F\u22efcentroid distances of 2.925\u20133.984\u2005\u00c5) inter\u00adactions are found, leading to a three-dimensional architecture. The RuII atom is coordinated in a distorted octa\u00adhedral geometry, particularly manifested by the Namine\u2014Ru\u2014Namine angle of 153.79\u2005(10)\u00b0. The counter-charge is provided by two hexa\u00adfluorido\u00adphosphate anions and the asymmetric unit is completed by aceto\u00adnitrile solvent mol\u00adecules of crystallization. Disorder was observed for both the hexa\u00adfluorido\u00adphosphate anions as well as the aceto\u00adnitrile solvate mol\u00adecules, with occupancies for the major moieties of 0.801\u2005(6) for one of the PF6 anions, and a shared occupancy of 0.9215\u2005(17) for the second PF6 anion and a partially occupied aceto\u00adnitrile mol\u00adecule. A second CH3CN mol\u00adecule is fully occupied, but 1:1 disordered across a crystallographic inversion center.In the title compound, [Ru(C The microwave-assisted synthesis of the precursor [Ru(L\u2013N4Me2)]2+, starting from [Ru(DMSO)4Cl2] and L\u2013N4Me2, in an ethano\u00adlic solution finished within 30\u2005min. It is not only fast, but also reproducible with only few byproducts, and hence requires no labor-intensive workup. Moreover, using the C2v symmetric macrocycle rather than bi\u00adpyridine-type ligands avoids the formation of mono- and dinuclear complexes with multiple stereoisomeric forms -pyridino\u00adphane (L\u2013N4H2) as a new ligand for RuII. Herein, we present the structure of the complex [Ru(L\u2013N4H2)tape](PF6)2, , obtained as its aceto\u00adnitrile solvate.Heteroaromatic ligands with more than three fused rings are commonly called large-surface ligands. Such ligands have attracted attention due to their use as connecting building blocks for supra\u00admolecular assemblies. If large-surface ligands feature more than one ligand donor site, connection between neighboring complexes can be realized through normal metal coordination (Ishow 4H2)tape]2+ in [Ru(C14H16N4)(C16H8N4](PF6)2\u00b71.422CH3CN. The Ru\u2014N bond lengths formed by the tape ligand (Table\u00a014Me2)tape]2+ \u00b0] from the idealized value of 180\u00b0 is slightly larger than for analogous ruthenium L\u2013N4Me2 complexes 2+, the dimers are also connected through bifurcated hydrogen bonds between one of the two L\u2013N4H2 ligand amine protons and both nitro\u00adgen atoms of the non-coordin\u00adating tape ligand \u03b1,\u03b1\u2032-di\u00adimine unit of the second complex cation of the dimer. In the crystal structure, these additional hydrogen bonds result in a short Ru\u22efRu distance of 8.8306\u2005(2)\u2005\u00c5, a tape ligand centroid\u2013centroid distance of 3.49\u2005(2)\u2005\u00c5 and an angle of 13.7 (1.4)\u00b0 between the ring normal and the centroid-to-centroid vector. Therefore, the \u03c0\u2013\u03c0 stacking motif can be described as parallel-displaced, but near to face-to-face -pyridino\u00adphane, see Brietzke, Mickler, Kelling, Schilde et al. tape](PF6)2 was synthesized as reported for [Ru(L\u2013N4Me2)tape](PF6)2 instead of L\u2013N4Me2. A yield of 44% was obtained; m.p. > 573\u2005K. 1H NMR = (MeCN\u2013d3): \u03b4 = 8.69 , 8.56 , 8.01 , 7.77 , 7.72 , 7.65 , 5.6 , 4.83 , 4.47 p.p.m. 13C NMR = (MeCN-d3): \u03b4 = 160.0 (C2 + C6), 152.4 (Ce), 150.28 (Cd), 150.24 (Ca), 145.7 (Cf), 138.9 (C4), 136.7 (Cb\u2032), 123.5 (Cb), 122.7 (C3 + C5), 122.0 (Cc), 119.3 (Ce\u2032), 64.8 (CH2) p.p.m. ESI=-MS: calculated for [M\u2013PF6]+ 743.0809; found 743.0778.The syntheses of the ligands L\u2013N4H2)tape](PF6)2. The solution was filled into a test tube, which was placed into a diethyl ether-containing bottle. Dark-green crystals began to form at ambient temperature within a few days.Crystals suitable for X-ray structure analysis were obtained by vapor diffusion of diethyl ether into a saturated aceto\u00adnitrile solution of [Ru(L\u2013N6 anions were refined as disordered over one major and one minor moiety each. The geometry of the minor moieties were each restrained to be similar to that of the major moieties (within an estimated standard deviation of 0.02\u2005\u00c5). The minor moieties were subjected to a rigid bond restraint , and the anisotropic displacement parameters of the major and minor phospho\u00adrus atoms were each constrained to be identical. Associated with the major moiety of the PF6 anion of P1 is an aceto\u00adnitrile mol\u00adecule that is absent for the minor moiety. Subject to the restraints and constraints used, the occupancy ratios refined to 0.9215\u2005(17) to 0.0785\u2005(17) for the PF6 units of P1A and P1B, and to 0.801\u2005(6) and 0.199\u2005(6) for those of P2A and P2B.Disorder was observed for both the hexa\u00adfluorido\u00adphosphate anions as well as the aceto\u00adnitrile solvate mol\u00adecules. Both PFUij components to their neighbors closer than 2\u2005\u00c5, including those of symmetry-related atoms .A second aceto\u00adnitrile mol\u00adecule is disordered across a crystallographic inversion center, with substantial overlap for the two carbon atoms of symmetry-related mol\u00adecules. The geometry of the mol\u00adecule was restrained to be similar to that of the first aceto\u00adnitrile mol\u00adecule, and the ADPs of its C and N atoms were restrained to be have similar 3), 0.99 (CH2), 0.95 (Carom), N\u2014H = 1.0\u2005\u00c5, and with Uiso(H) = 1.2Ueq(C) with the exception of methyl hydrogen atoms, which were refined with Uiso(H) = 1.5Ueq(C).All hydrogen atoms connected to C and N atoms were placed in their expected calculated positions and refined as riding with C\u2014H = 0.98 global, I. DOI: 10.1107/S1600536814021060/zl2602Isup2.hklStructure factors: contains datablock(s) I. DOI: 1025408CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Water mol\u00adecules play an important role in the structure connectivity.In a novel one-dimensional hybrid compound, the inorganic part contains zigzag chains formed by corner-sharing [SbCl catena-poly[[[bis\u00ad [tetra\u00adachlorido\u00adanti\u00admonate(III)]-\u03bc-chlorido-[tetra\u00adchlorido\u00adanti\u00admonate(III)]-\u03bc-chlorido]] monohydrate], {(C6H14N2)2[Sb2Cl10]\u00b7H2O}n, is self-assembled into alternating organic and inorganic layers parallel to the bc plane. The anionic inorganic layer consists of infinite zigzag chains of corner-sharing [SbCl6]3\u2212 octa\u00adhedra running along the b axis. The organic part is made up of 1,4-diazo\u00adniabi\u00adcyclo\u00ad[2.2.2]octane dications (dabcoH22+). The water mol\u00adecules in the structure connect inorganic and organic layers. Hydrogen-bonding inter\u00adactions between the ammonium groups, water mol\u00adecules and Cl atoms ensure the structure cohesion.The hybrid title compound, Ra)n+MbX3b+na} are able to combine desirable characteristics from both types of constituents into a mol\u00adecular scale composite. These hybrids have been extensively studied for their excitonic and magneto-optical properties. In recent years, a significant number of organic\u2013inorganic hybrid materials based on anti\u00admony\u2013halide units have been studied. Six-coordinate anti\u00admony halides can arrange themselves in three-, two- or one-dimensional networks through sharing halides in the SbX6 octa\u00adhedra, separated by organic cations 2[Sb2Cl10]\u00b7H2O, (I)2)2+ dications, a corner sharing bi-octa\u00adhedron deca\u00adchlorido\u00addianti\u00admonate(III) anion and one crystallization water mol\u00adecule. The cations are labeled Cat1 (containing atoms N1 and N2) and Cat2 (containing N3 and N4) and the atomic numbering scheme is shown in Fig.\u00a01The asymmetric unit of the new chlorido\u00adanti\u00admonate(III) compound, 2)2+ cations located in the holes around the corner-sharing octa\u00adhedra. The layers are stacked along the a axis and water mol\u00adecules connect the organic and inorganic components ts Fig.\u00a02.6]3\u2212 and [Sb2Cl6]3\u2212) joined by the Cl2 ion. Both octa\u00adhedra are severely distorted with Sb\u2014Cl bond lengths lying in the range of 2.5233\u2005(18)\u20133.073\u2005(2)\u2005\u00c5 for the bridging ones and 2.4277\u2005(15)\u20132.8233\u2005(17)\u2005\u00c5 for the terminal ones. The two bridging halides (Cl2 and Cl4) connecting the central octa\u00adhedron to its neighbours are related cis, leading to zigzag chain of corner-sharing [SbCl6]3\u2212 octa\u00adhedra running along the b axis is Fig.\u00a03.P63/m) 2+ cations exhibit deviations from ideal Dh3 symmetry. The observed lowering symmetry is probably due to the distortion of the (dabcoH2)2+ cation and can be related to the complex hydrogen-bond network linking the mol\u00adecular components .It is worth noting that at room temperature the DABCO mol\u00adecule crystallizes in the hexa\u00adgonal system (Pna21). These are two indispensable conditions making this phase a potential promising candidate for non-linear optical (NLO) behaviour as is the case for the well-known KTiOPO4 (KTP) and equivalent efficient NLO materials.The studied compound crystals are transparent and the structure is noncentrosymmetric (2)2+ cations and two water mol\u00adecules via hydrogen bonds \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0\u2212\u00a0x\u00a0+\u00a0y\u00a0\u2212\u00a0z\u00a0\u2212\u00a0via Cl1\u22efH2iii\u2014N2iii, Cl3\u22efH2iii\u2014N2iii and Cat2 by Cl3\u22efH4ii\u2014N4ii 3\u2212 octa\u00adhedra via O\u2014H13A\u22efCl5 and O\u2014H13B\u22efCl9iii \u00b72H2O containing isolated [Sb2Cl10]4\u2212 double octa\u00adhedra, (dabcoH2)2+ cations and water mol\u00adecules 2NH(CH2)2NH3]2+, see: Bujack & Angel and DABCO (0.5\u2005mmol) was dissolved in a hydro\u00adchloric aqueous solution and stirred for several minutes at 353\u2005K. Colourless crystals suitable for X-ray diffraction analysis were obtained by slow evaporation at room temperature after two weeks.A mixture of SbClSHELXL I, New_Global_Publ_Block. DOI: 10.1107/S2056989015007379/vn2091Isup2.hklStructure factors: contains datablock(s) I. DOI: 943047CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure of 2,2\u2032\u2032-bis\u00ad-1,1\u2032:4\u2032,1\u2032\u2032-terphenyl as the tris\u00ad(tri\u00adethyl\u00adamine) solvate, the diol host mol\u00adecule possesses a \u2018folded\u2019 mol\u00adecular conformation with inversion symmetry. Two of the three solvent mol\u00adecules form O\u2014H\u22efN hydrogen bonds and the third one forms C\u2014H\u22efN hydrogen bonds with the host compound. 44H26Cl4O2\u00b73C6H15N, the asymmetric part of the unit cell comprises two halves of the diol mol\u00adecules, 2,2\u2032\u2032-bis\u00ad-1,1\u2032:4\u2032,1\u2032\u2032-terphenyl, and three mol\u00adecules of tri\u00adethyl\u00adamine, i. e. the diol mol\u00adecules are located on crystallographic symmetry centres. Two of the solvent mol\u00adecules are disordered over two positions [occupancy ratios of 0.567\u2005(3):0.433\u2005(3) and 0.503\u2005(3):0.497\u2005(3)]. In the diol mol\u00adecules, the outer rings of the 1,1\u2032:4\u2032,1\u2032\u2032-terphenyl elements are twisted with reference to their central arene ring and the mean planes of the fluorenyl moieties are inclined with respect to the terphenyl ring to which they are connected, the latter making dihedral angles of 82.05\u2005(8) and 82.28\u2005(8)\u00b0. The presence of two 9-fluoren-9-ol units attached at positions 2 and 2\u2032\u2032 of the terphenyl moiety induces a \u2018folded\u2019 geometry which is stabilized by intra\u00admolecular C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions, the latter formed between the fluorenyl units and the central ring of the terphenyl unit [centroid\u2013centroid distances = 3.559\u2005(1) and 3.562\u2005(1)\u2005\u00c5]. The crystal is composed of 1:2 complex units, in which the solvent mol\u00adecules are associated with the diol mol\u00adecules via O\u2014H\u22efN hydrogen bonds, while the remaining solvent mol\u00adecule is linked to the host by a C\u2014H\u22efN hydrogen bond. The given pattern of inter\u00admolecular inter\u00adactions results in formation of chain structures extending along [010].In the title solvate, C Moreover, the location of the central ring of the terphenyl unit between the fluorenyl units [ring centroid distances = 3.559\u2005(1) and 3.562\u2005(1)\u2005\u00c5] indicate the presence of \u03c0\u2013\u03c0 stacking inter\u00adactions with respect to the inner ring (A or A\u2032) Fig.\u00a02 of the td(H\u22efN) 1.91\u20131.95\u2005\u00c5] (Table\u00a01via C\u2014H\u22efO hydrogen bonding , giving an overall chain structure extending along [010] -1,1\u2032:2\u2032,1\u2032\u2032:4\u2032\u2032,1\u2032\u2032\u2032:2\u2032\u2032\u2032,1\u2032\u2032\u2032\u2019-quinquephenyl bis\u00ad(di\u00adethyl\u00adamine) clathrate bis\u00ad(propan-1-ol) clathrate bis\u00ad(but\u00adan-1-ol) clathrate bis\u00ad(ethanol) clathrate bis\u00ad(propan-1-ol) clathrate bis\u00ad(di\u00adethyl\u00adamine) clathrate bis\u00ad(butan-1-ol) clathrate to a cold solution (195\u2005K) of 2,2\u2032\u2032-di\u00adiodo-1,1\u2032:4\u2032,1\u2032\u2032-terphenyl in 20\u2005ml of dry THF. After 45\u2005min of stirring, 4,4\u2032-di\u00adchloro\u00adbenzo\u00adphenone , dissolved in 4\u2005ml benzene and 15\u2005ml THF, was added. The colourless reaction mixture was warmed to room temperature and stirred for 4\u2005h. The solution was extracted twice with diethyl ether. The combined organic extracts were washed with water, dried over anhydrous sodium sulfate and concentrated under reduced pressure. Colourless crystals were isolated by recrystallization from hexane (yield: 7.0%). M.p. 543\u2013546\u2005K; ESI\u2013MS [M + H]\u2212m/z 731.3. IR (KBr) \u03bd (cm\u22121) 3547, 3056, 3025, 1913, 1641, 1591, 1575, 1489, 1331, 1182, 1157, 1097, 1014, 919, 903, 840, 761. 1H NMR : \u03b4 = 2.84 , 6.75 , 7.09 , 7.11 , 7.22 , 7.26 , 7.32 . 13C NMR : \u03b4 = 82.68 (C-OH), 126.89, 127.43, 128.10, 129.11, 129.33, 129.83, 133.40, 140.24, 141.01, 144.06, 145.58 (Ar-C). EA calculated for C44H30O2Cl4: C 72.1, H 4.1%; found: C 72.2, H 4.4%. Crystals of (IIa) suitable for X-ray diffraction were obtained from a solution of (II) in tri\u00adethyl\u00adamine upon slow evaporation of the solvent at room temperature.The unsolvated compound (II) was prepared by addition of a solution of Uiso(H) = 1.2Ueq(C) for aromatic and methyl\u00adene, with C\u2014H = 0.98 and O\u2014H = 0.84\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for methyl and hy\u00addroxy groups, respectively. Two mol\u00adecules of tri\u00adethyl\u00adamine are each disordered over two positions with occupancy ratios of 0.567\u2005(3):0.433\u2005(3) and 0.503\u2005(3):0.497\u2005(3). They were modelled with restrained bond lengths based on average values of 1.47\u2005(1)\u2005\u00c5 for N\u2014C and 1.53\u2005(1)\u2005\u00c5 for C\u2014C bonds.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015018824/zs2345sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989015018824/zs2345Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015018824/zs2345Isup3.cmlSupporting information file. DOI: 1430018CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Cations and anions are linked through C\u2014H\u22efO hydrogen bonds.The asymmetric unit of the title compound consists of one [Eu(C 3H7NO)8][PMo12O40], the asymmetric unit comprises one \u03b1-Keggin-type [PMo12O40]3\u2212 polyoxidometalate anion and one distorted dodeca\u00adhedral [Eu(C3H7NO)8]3+ complex cation. In the crystal, the isolated polyoxidometalate anions are packed into hexa\u00adgonally arranged rows extending parallel to [001]. The complex cations are situated between the rows and are linked to the neighbouring anions through weak C\u2014H\u22efO hydrogen-bonding inter\u00adactions, leading to the formation of a three-dimensional network structure.In the title salt, [Eu(C The latter species are fused together by sharing corner atoms (Ob) and consist of three MoO6 octa\u00adhedra condensed in a triangular arrangement by sharing edges (Oc). There is also a terminal oxygen atom (Od) in every MoO6 octa\u00adhedron. The P\u2014O bond lengths range from 1.521\u2005(5)\u2005\u00c5 to 1.536\u2005(4)\u2005\u00c5 and the Mo\u2014O bond lengths from 1.690\u2005(5)\u2005\u00c5 to 2.438\u2005(4)\u2005\u00c5. The O\u2014P\u2014O angles [109.1\u2005(2)\u2013109.8\u2005(3)\u00b0] indicate only a slight distortion of the central PO4 tetra\u00adhedron. The EuIII cation is coordinated by eight di\u00admethyl\u00adformamide ligands through their oxygen atoms with Eu\u2014O distances from 2.369\u2005(5) to 2.416\u2005(6)\u2005\u00c5. These values are comparable to those of related oxido-europium(III) species, e.g for the [Eu(thd)3(DMF)2] complex with Eu\u2014O = 2.494\u2005(5)\u20132.442\u2005(5)\u2005\u00c5 8]3+ cation is linked to four neighbouring \u03b1-Keggin-type [PMo12O40]3\u2212 anions through C\u2014H\u22efO hydrogen-bonding inter\u00adactions between the methyl groups of the DMF ligands and the terminal-oxygen (Od) and the bridging-oxygen atoms of the [PMo12O40]3\u2212 anions distances are between 3.174\u2005(10) and 3.541\u2005(11)\u2005\u00c5 while the C\u22efO distances are between 3.289\u2005(11) and 3.473\u2005(12)\u2005\u00c5. In the crystal packing, the POM anions are packed into hexa\u00adgonally arranged rows extending parallel to [001] with the [Eu(DMF)8]3+ cations located between the rows 4N)4H3][PMo11O39] was prepared using a literature method and isonicotinic acid (C6H5NO2) were dissolved in 10\u2005ml of di\u00admethyl\u00adformamide. This solution was added dropwise to a yellow di\u00admethyl\u00adformamide solution of [(C4H9)4N)4H3][PMo11O39] (0.33\u2005mmol in 10\u2005ml). The mixture was heated under stirring for 1\u2005h at 333\u2005K. Single crystals of the title compounds were obtained by slow diffusion of 2-propanol through the di\u00admethyl\u00adformamide solution. UV\u2013vis spectrum in di\u00admethyl\u00adformamide: \u03bbmax (nm) 315 and 205.The starting material [, \u03bdas(Mo=Od), \u03bdas(Mo\u2014Ob\u2014Mo) and \u03bdas(Mo\u2014Oc\u2014Mo) appear at 1065, 951, 885 and 974\u2005cm\u22121, respectively (rocking vibration), \u03b4a(CH3), \u03b4s(CH3) and \u03bd(C-H) of the di\u00admethyl\u00adformamide ligand Uiso(H) = 1.2Ueq(C) for methine groups and C\u2014H = 0.96\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for methyl groups. The refined Flack parameter I. DOI: 10.1107/S2056989016003546/wm5274Isup2.hklStructure factors: contains datablock(s) I. DOI: 1456619CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Molecules in both polymorphs of the title compound display deviations from planarity owing to crystal packing effects. 12H11N3O2, is a second monoclinic polymorph of the previously reported monoclinic form [Akhmad Aznan et al. to 26.24\u2005(19)\u00b0. The geometry-optimized structure [B3LYP level of theory and 6\u2013311\u2005g+ basis set] has the same features except that the entire mol\u00adecule is planar. In the crystal, the three-dimensional architecture is consolidated by a combination of C\u2014H\u22efO, C\u2014H\u22ef\u03c0, nitro-N\u2014O\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions [inter-centroid distances = 3.649\u2005(2)\u20133.916\u2005(2)\u2005\u00c5].The title compound, C al. 2010. Acta Cr Similar conformations were observed for the two independent mol\u00adecules in the previously reported P21/c polymorph inter\u00adaction to close the 10-membered {\u22efHC2NO}2 synthon is a little longer that the standard distance criteria incorporated in PLATON inter\u00adactions ], and toluene C18\u2013C23 and N8-pyridine rings . As summarized in Table\u00a01via meth\u00adyl\u2013toluene C\u2014H\u22ef\u03c0 inter\u00adactions, and layers are connected into a three-dimensional architecture \u22ef(C18\u2013C23) = 3.916\u2005(2)\u2005\u00c5, with an angle of inclination of 11.04\u2005(19)\u00b0, and inter\u00adcentroid distance for (C6\u2013C11)\u22ef(N5/C13\u2013C17) = 3.913\u2005(2)\u2005\u00c5, with an angle of inclination of 13.44\u2005(19)\u00b0 and symmetry operation .Globally, the crystal packing features alternating layers of mol\u00adecules that stack along the s Table\u00a01. Ten-memns Fig.\u00a03. These cns Fig.\u00a04. The \u03c0\u2013\u03c0re Fig.\u00a05 via weakN-(3-chloro\u00adphen\u00adyl)-3-nitro\u00adpyridin-2-amine hydrogen bond, the coplanarity of the nitro group and pyridine ring, and a conrotatory twist of the two rings, i.e. dihedral angles of 9.88\u2005(5) and 84.77\u2005(10)\u00b0, respectively. Finally, the structure of the all-phenyl analogue, 2-nitro\u00addiphenyl\u00adamine, has been reported -3-nitro\u00adpyridin-2-amine , prepared according to the literature procedure of Akhmad Aznan et al. set at 1.2Ueq(C). N-bound H atoms were located in a difference Fourier map but were refined with a distance restraint of N\u2014H = 0.88\u00b10.01\u2005\u00c5 and with Uiso(H) set at 1.2Ueq(N). In the absence of significant anomalous scattering effects, 4208 Friedel pairs were averaged in the final refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814012227/hb0011sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S1600536814012227/hb0011Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814012227/hb0011Isup3.cmlSupporting information file. DOI: 1004278CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The furan and one of the phenanthrene groups are fused in an almost coplanar arrangement [dihedral angle = 5.14\u2005(8)\u00b0] and the furan unit makes dihedral angles of 70.27\u2005(11) and 57.58\u2005(8)\u00b0 with the planes of the phenyl and the second phenanthrene group, respectively. In the crystal, neighbouring mol\u00adecules are connected via two inter\u00admolecular hydrogen-bonding inter\u00adactions (O\u2014H\u22efO and C\u2014H\u22efO) towards the carbonyl O atom with donor\u2013acceptor distances of 2.824\u2005(2) and 3.277\u2005(3)\u2005\u00c5, creating an inversion dimer. A non-classical C\u2014H\u22efCl inter\u00adaction [3.564\u2005(2)\u2005\u00c5] and three C\u2014H\u22ef\u03c0 inter\u00adactions, with C\u22ef\u03c0 distances of 3.709\u2005(3), 3.745\u2005(2) and 3.628\u2005(3)\u2005\u00c5, connect the mol\u00adecules, forming a three-dimensional supra\u00admolecular architecture in the solid state.In the title compound, C The cenb-axis direction -furan\u00adone [2-(4-chloro\u00adphen\u00adyl)-2-hy\u00addroxy-1-oxa\u00adcyclo\u00adpenta\u00ad[l]phenanthren-3-one] (4) (65%), which was purified by recrystallization from a mixture of methanol and di\u00adchloro\u00admethane (2:1 v/v). The title compound (3) was the minor product formed along with (4) during the reaction . Diffraction-quality single crystals were generated by slow evaporation from methanol. Yield 1.90\u2005g (14%); m.p. 459\u2005K; IR : 3374 (OH), 1591 (C=O) cm\u22121; 1H NMR (CDCl3): \u03b4 8.79\u20137.26 , 8.69 ; MS: m/z 548 (M+). Analysis calculated for C37H21ClO3: C 80.94, H 3.86%; found: C 80.82, H 3.66%.A mixture of phenanthrene\u00adquinone (1) , 4-chloro\u00adaceto\u00adphenone (2) and powdered potassium hydroxide (1\u2005g) in methanol (30\u2005ml) was stirred at 333\u2005K for 4\u2005h and then kept in a refrigerator for 48\u2005h. The main product obtained was a 3 = 1.2Ueq(C). The phenanthroline atom H3 was located from a difference Fourier map and refined with a distance restraint of O\u2014H = 0.86\u2005(1)\u2005\u00c5. The reflection 101 was omitted owing to bad agreement. Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814022338/zl2597sup1.cifCrystal structure: contains datablock(s) 3. DOI: 10.1107/S1600536814022338/zl25973sup2.hklStructure factors: contains datablock(s) 3. DOI: 1024475CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Page 1: In the Acknowledgements: \u201cproject ID RFMEFI57614X0061\u201d should read \u201cproject ID RFMEFI60414X0022.\u201dVolume 2, no. 6,"} +{"text": "The torsion angles indicate a non-helical conformation of the mol\u00adecule.In a de\u00adhydro\u00adamino acid with a C=C bond between the \u03b1- and \u03b2-C atoms, the amino acid residues are linked 11H17BrN2O5, is a de\u00adhydro\u00adamino acid with a C=C bond between the \u03b1- and \u03b2-C atoms. The amino acid residues are linked trans to each other and there are no strong intra\u00admolecular hydrogen bonds. The torsion angles indicate a non-helical conformation of the mol\u00adecule. The dipeptide folding is influenced by an inter\u00admolecular N\u2014H\u22efO hydrogen bond and also minimizes steric repulsion. In the crystal, mol\u00adecules are linked by strong N\u2014H\u22efO hydrogen bonds, generating (001) sheets. The sheets are linked by weak C\u2014H\u22efO and C\u2014H\u22efBr bonds and short Br\u22efBr [3.4149\u2005(3)\u2005\u00c5] inter\u00adactions.The title compound, C This reaction proceeds in two steps, namely by halogenation of de\u00adhydro\u00adamino acids, which gives \u03b1-bromo-imines, followed by tautomerization to the desired products upon treatment with an amine is 175.79\u2005(16)\u00b0, while \u03c93 (O5\u2014C6\u2014N8\u2014C9) is 176.12\u2005(15)\u00b0. There are no strong intra\u00admolecular hydrogen bonds. The values of the \u03d52,3 and \u03c82,3 angles corresponds to a non-helical conformation \u00b0], which may correspond to the presence of the bromine atom in the structure. The other angles are normal, as the backbone of the mol\u00adecule is folded to minimize steric repulsion. The Boc group features two short intra\u00admolecular C\u2014H\u22efO contactsThe mol\u00adecular structure of the title compound, (I)A\u22efO17i and N12\u2014H12A\u22efO7ii) and one weak accompanying C9\u2014H9A\u22efO11i hydrogen bonds \u2212x\u00a0+\u00a03, \u2212y, \u2212z\u00a0+\u00a01]. The sheets are connected to each other by weak C14\u2014H14A\u22efO11iii and C19\u2014H19B\u22efBr15iii hydrogen bonds and one Br\u22efBriv [3.4149\u2005(3)\u2005\u00c5] halogen bond \u2212x\u00a0+\u00a03, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01].In the crystal, mol\u00adecules form two strong twin N\u2014H\u22efO of Boc\u2013Gly\u2013\u0394Ala\u2013OMe was dissolved in 2.5\u2005ml of di\u00adchloro\u00admethane and cooled to 193\u2005K. Then, bromine 0.027\u2005ml (0.5\u2005mM) was added. The solution was stirred over 10 minutes followed by addition of tri\u00adethyl\u00adamine 0.210\u2005ml (1.5\u2005mM). After 15 minutes, the mixture was quenched with 20\u2005ml of saturated aqueous NaHCO3 and warmed to room temperature. The product was extracted by di\u00adchloro\u00admethane (3 \u00d7 15\u2005ml). The organic layer was washed with brine (3 \u00d7 10\u2005ml) and dried over anhydrous Na2SO4. Evaporation of the solvent at reduced pressure gave 0.119\u2005g (0.35\u2005mM) of crude product (70% yield). Recrystal\u00adlization was performed from mixtures of diethyl ether/ethyl acetate\u00ad(2:1)/hexane solvents, yielding irregular colourless crystals. It is worth noting that in the case of our study, the formation of only the Z isomer was observed while in the preceding paper, the bromination of de\u00adhydro\u00adalanine-containing compound gave the E isomer. 1H NMR \u03b4 1.38 , 3.67 , 3.69 , 7.05 , 7.30 , 9.63 . 13C NMR \u03b4 28.21, 42.79, 52.54, 78.12, 113.26, 132.88, 155.80, 162.63, 168.80. Melting point = 386\u2013388\u2005K.Boc\u2013Gly\u2013\u0394Ala and its methyl ester were prepared according to the methodology described by Makowski al. 1985 and Coss al. 2008. The \u03b2-bUiso (H) = 1.5Ueq(C); for N atoms, N\u2014H = 0.86\u2005\u00c5 and Uiso (H) = 1.2Ueq(C); for secondary C atoms, C\u2014H = 0.97\u2005\u00c5 and Uiso (H) = 1.2Ueq(C), with no refinement of their parameters.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814025677/hb7312sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S1600536814025677/hb7312Isup2.hklStructure factors: contains datablock(s) I. DOI: 1035539CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the complex cation, the SbV atom lies on an inversion center and is octa\u00adhedrally coordinated by four N atoms from a macrocyclic tetra\u00adphenyl\u00adporphyrinate ligand and two chloride ions. The complex cation has approximately a planar core with a maximum deviation of 0.018\u2005(5)\u2005\u00c5 from the porphyrin mean plane. The average Sb\u2014N distance is 2.062\u2005(11)\u2005\u00c5, while the Sb\u2014Cl distance is 2.355\u2005(1)\u2005\u00c5. The SbV atom of the anion is also located on an inversion center. The [SbCl6]\u2212 octa\u00adhedron exhibits rhombic distortion characterized by the Sb\u2014Cl bond lengths . In the crystal, the cations and anions are linked C\u2014H\u22ef Cl hydrogen bonds, forming a layer parallel to (211).The asymmetric unit of the title compound, [Sb(C Cl2][SbCl6] = 0.065wR(F2) = 0.173S = 1.134749 reflections266 parametersH-atom parameters constrainedmax = 2.55 e \u00c5\u22123\u0394\u03c1min = \u22121.45 e \u00c5\u22123\u0394\u03c1COLLECT used to solve structure: SIR2004 I, New_Global_Publ_Block. DOI: 10.1107/S1600536814012653/is5356Isup2.hklStructure factors: contains datablock(s) I. DOI: 1006075CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III ion in the title cationic complex is coordinated by four N atoms from the macrocyclic ligand, one water mol\u00adecule and one chloride in a cis geometry, displaying a distorted octa\u00adhedral environment. The crystal packing is stabilized by N\u2014H\u22efCl, O\u2014H\u22efCl and O\u2014H\u22efO hydrogen bonds.The Cr cis-[CrCl(cycb)(H2O)][ZnCl4]\u00b73H2O , has been determined from synchrotron data. In the complex cation, the CrIII ion is bound by four N atoms from the tetra\u00addentate cycb ligand, a chloride ion and one water mol\u00adecule in a cis arrangement, displaying a distorted octa\u00adhedral coordination geometry. The distorted tetra\u00adhedral [ZnCl4]2\u2212 anion and three additional water mol\u00adecules remain outside the coordination sphere. The Cr\u2014N(cycb) bond lengths are in the range of 2.0837\u2005(14) to 2.1399\u2005(12)\u2005\u00c5 while the Cr\u2014Cl and Cr\u2014(OH2) bond lengths are 2.2940\u2005(8) and 2.0082\u2005(13)\u2005\u00c5, respectively. The crystal packing is stabilized by hydrogen-bonding inter\u00adactions between the N\u2014H groups of the macrocyclic ligand, the O\u2014H groups of the water mol\u00adecules and the Cl atoms of the tetra\u00adchlorido\u00adzincate anion, leading to the formation of a three-dimensional network.The structure of the title compound, C-meso or racemic-5,5,7,12,12,14-hexa\u00admethyl-1,4,8,11-tetra\u00adaza\u00adcyclo\u00adtetra\u00addecane (cyca and cycb) ligands are known to exist in trans or cis octa\u00adhedral coordination geometries when combined with two auxiliary ligands (OH2)]ZnCl4\u00b73H2O, (I)Chromium(III) complexes containing III ion with an O1A\u2014Cr1A\u2014Cl1A bond angle of 85.74\u2005(4)\u00b0. The rest of the coordination sites are occupied by four nitro\u00adgen atoms of the tetra\u00addentate macrocyclic cycb ligand, giving rise to a distorted octa\u00adhedral coordination sphere.In the mol\u00adecular structure of the complex cation, there is one chlorine atom and one water mol\u00adecule coordinating the Crb ligand is folded about the N2A\u2014Cr1A\u2014N4A line and is in its most stable cis-V conformation bond lengths are in the range 2.0837\u2005(14) to 2.1399\u2005(12)\u2005\u00c5, in good agreement with those observed in cis-[Cr(OH)2(cycb)]ClO4\u00b72H2O ClO4\u00b7H2O Br\u00b7H2O Cl ClO4\u00b70.5H2O (NO3)2 Cl (NO3)2 (OH2)](ClO4)2\u00b7H2O and 172.43\u2005(5)\u00b0, respectively. The angles N1A\u2014Cr1A\u2014N2A and N3A\u2014Cr1A\u2014N4A are 87.01\u2005(5) and 87.77\u2005(5)\u00b0, reflecting the distorted octa\u00adhedral coordination sphere. The tetra\u00adhedral [ZnCl4]2\u2212 anion and three additional water mol\u00adecules remain outside the coordination sphere of CrIII. The complex anion is distorted due to its involvement in hydrogen-bonding inter\u00adactions. Zn\u2014Cl bonds in the anion span a range from 2.2569\u2005(7) to 2.3131\u2005(8)\u2005\u00c5, and the Cl\u2014Zn\u2014Cl angles from 106.02\u2005(4) to 111.49\u2005(3)\u00b0.The cycon Fig.\u00a01. The Cr\u2014Extensive hydrogen-bonding inter\u00adactions occur in the crystal structure Table\u00a01. The supIII complexes involving the macrocyclic rac-5,5,7,12,12,14-hexa\u00admethyl-1,4,8,11-tetra\u00adaza\u00adcyclo\u00adtetra\u00addecane ligand. The crystal structures of cis-[Cr(OH)2(cycb)]ClO4\u00b72H2O 2(cycb)]ClO4\u00b7H2O (cycb)]Br\u00b7H2O 2(cycb)]Cl (cycb)]ClO4\u00b70.5H2O (OH2)](NO3)2 (cyca)(OH2)](ClO4)2\u00b7H2O (OH2)]2+ cationic complex with any anion was found, although the preparation of cis-[CrCl(cycb)(OH2)](ClO4)2\u00b70.4HClO4\u00b73H2O has been reported ]Cl\u00b7H2O was prepared according to literature procedures ]Cl\u00b7H2O (0.07\u2005g) was dissolved in 4\u2005mL of 0.01 M HCl at 353\u2005K and the 1\u2005mL of 6 M HCl containing 0.15\u2005g of solid ZnCl2 were added to this solution. The mixture was refluxed for 30\u2005min and then cooled to room temperature. The resulting solution was filtered and the filtrate was allowed to stand at room temperature for one day to afford purple crystals of compound (I)All chemicals were reagent grade materials and used without further purification. The starting material, Uiso(H) values of 1.2 or 1.5 \u00d7 Ueq of the parent atoms. The hydrogen atoms of water mol\u00adecules were located in difference maps restrained with O\u2014H = 0.84\u2005\u00c5 using DFIX and DANG commands.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015015212/wm5196sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015015212/wm5196Isup2.hklStructure factors: contains datablock(s) I. DOI: 1419197CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The structures of two isomeric compounds of iso\u00adquinoline with 3-chloro-2-nitro\u00adbenzoic acid and 4-chloro-2-nitro\u00adbenzoic acid have been determined at 190\u2005K. In each compound, the acid and base mol\u00adecules are held together by a short hydrogen bond between a carb\u00adoxy O atom and a base N atom. In the hydrogen-bonded unit of the former, the H atom is disordered over two positions, while in the latter, an acid\u2013base inter\u00adaction involving H-atom transfer occurs and the H atom is located at the N site. 9H7.3N\u00b7C7H3.7ClNO4, (I), and C9H8N\u00b7C7H3ClNO4, (II), of iso\u00adquinoline with 3-chloro-2-nitro\u00adbenzoic acid and 4-chloro-2-nitro\u00adbenzoic acid, the two components are linked by a short hydrogen bond between a base N atom and a carb\u00adoxy O atom. In the hydrogen-bonded unit of (I), the H atom is disordered over two positions with N and O site occupancies of 0.30\u2005(3) and 0.70\u2005(3), respectively, while in (II), an acid\u2013base inter\u00adaction involving H-atom transfer occurs and the H atom is located at the N site. In the crystal of (I), the acid\u2013base units are connected through C\u2014H\u22efO hydrogen bonds into a tape structure along the b-axis direction. Inversion-related adjacent tapes are further linked through \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.6389\u2005(7)\u20133.7501\u2005(7)\u2005\u00c5], forming a layer parallel to (001). In the crystal of (II), the acid\u2013base units are connected through C\u2014H\u22efO hydrogen bonds into a ladder structure along the a-axis direction. The ladders are further linked by another C\u2014H\u22efO hydrogen bond into a layer parallel to (001).In each of the title isomeric compounds, C In the hydrogen-bonded unit, the iso\u00adquinoline ring system make dihedral angles of 54.12\u2005(15) and 71.89\u2005(5)\u00b0, respectively, with the carb\u00adoxy group and the benzene ring of the acid. In the acid mol\u00adecule, the benzene ring makes dihedral angles of 26.59\u2005(15) and 67.69\u2005(15)\u00b0, respectively, with the carb\u00adoxy and nitro groups.The mol\u00adecular structure of (II)i; Table\u00a01b-axis direction , Cg1\u22efCg2iv = 3.6674\u2005(7), Cg1\u22efCg3iii = 3.6637\u2005(7) and Cg1\u22efCg3iv = 3.6389\u2005(7)\u2005\u00c5, where Cg1, Cg2 and Cg3 are the centroids of the C1\u2013C6 benzene ring of the acid, and the N2/C8\u2013C10/C15/C16 rings of the base, respectively. Symmetry codes: (iii) \u2212x, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01; (iv) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01.]In the crystal of (I)on Fig.\u00a03. Adjacenon Fig.\u00a03, formingi and C13\u2014H13\u22efO3ii; Table\u00a02a-axis direction on Fig.\u00a05. AdjacenA search of the Cambridge Structural Database Uiso(H) = 1.5Ueq(N or O). The positional parameters were refined with bond restraints of O\u2014H = 0.84\u2005(2)\u2005\u00c5 and N\u2014H = 0.88\u2005(2)\u2005\u00c5. Atom H2 in (II)Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989014026152/lh5740sup1.cifCrystal structure: contains datablock(s) General, I, II. DOI: 10.1107/S2056989014026152/lh5740IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1036583, 1036582CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the components are linked by N\u2014H\u22efCl, N\u2014H\u22efO, O\u2014H\u22efCl and O\u2014H\u22efO hydrogen bonds, generating double layers propagating in (100).In the cation of the title hydrated mol\u00adecular salt, C As long ago as 1887, it was demonstrated -N--5-[amino]-2-hy\u00addroxy-4-oxo\u00adcyclo\u00adhexa-2,5-dien-1-iminium chloride, 4, which generates the zwitterion 5 when treated with base.In the course of our ongoing studies ] in the form of purple needles. This reaction must proceed via the elusive inter\u00admediate 6 which spontaneously hydrolyses. The first hydrolysis product should be inter\u00admediate 7. This contains a conjugated iminium salt and a vinyl\u00adogous amide, which must hydrolyse rapidly, possibly because of the stability of the acidic enol formed. It appears to be a rapid hydrolysis for an amide under mild conditions and so stabilization of a tetra\u00adhedral inter\u00admediate by the positive iminium salt might occur.By careful oxidation of the tetra\u00adhydro\u00adchloride salt of amine 6H7N2O2+ cation (r.m.s. deviation for the non-hydrogen atoms = 0.028\u2005\u00c5), a chloride counter-ion and a water mol\u00adecule of crystallization on Fig.\u00a01. Despitevia the C\u2014N and C\u2014C bonds between them. In terms of the \u2018oxygen side\u2019 of the cation, the C6\u2014O2 bond [1.320\u2005(4)\u2005\u00c5] is short for a C\u2014O single bond whereas C2\u2014O1 [1.227\u2005(4)\u2005\u00c5] is slightly lengthened for a nominal C=O double bond. This in combination with the C1\u2014C2 and C1\u2014C6 bond lengths again implies a degree of delocalization over these five atoms. However, the long C2\u2014C3 and C5\u2014C6 bonds imply little, if any, conjugation between the two delocalized components (O2/C6/C1/C2/O1 and N2/C5/C4/C3/N1) of the cation.The short C3\u2014C4 and C4\u2014C5 bonds correlate with the approximately equal C3\u2014N1 [1.320\u2005(4)] and C5\u2014N2 [1.306\u2005(4)\u2005\u00c5] bond lengths, which imply equal delocalization of the positive charge of the cation over atoms N1 and N2, mediated viz. N1\u2014H2n\u22efO1 and N2\u2014H4n\u22efO2 (Table\u00a01S(5) rings.The cation features two intra\u00admolecular N\u2014H\u22efO hydrogen bonds, 2 Table\u00a01, which bc-glide symmetry. Each link in the chain comprises two cations and two anions and In the crystal, the components are linked by N\u2014H\u22efCl, N\u2014H\u22efO, O\u2014H\u22efCl and O\u2014H\u22efO hydrogen bonds Table\u00a01. If the via the intra\u00admolecular N1\u2014H2n\u22efO1 hydrogen bond).When the cation and water mol\u00adecule are considered together, an [001] chain also arises Fig.\u00a03. The watw\u22efCl1 hydrogen bond providing the key link between the sheets. Overall, the chloride ion accepts four hydrogen bonds (three N\u2014H\u22efCl and one O\u2014H\u22efCl inter\u00adactions) in an irregular geometry.When all components are considered together, (100) double sheets result Fig.\u00a04, with thE)-N--5-[amino]-2-hy\u00addroxy-4-\u03bfxo\u00adcyclo\u00adhexa-2,5-dien-1-iminium chloride chloro\u00adform monosolvate and stirred at room temperature for 24\u2005h. The brown mixture was neutralized with NaHCO3 giving a brown or red precipitate, which was then extracted with CH2Cl2 (10 \u00d7 50\u2005ml). The yellow extracts were combined, deca\u00adnted, then stirred with methanol (50\u2005ml) containing five drops of conc. HCl(aq). The yellow solution turned purple. This was evaporated to dryness, then the product was dissolved in methanol (50\u2005ml) to yield a red solution and recrystallized by slow evaporation to leave the title compound as purple needles: m.p. > 473\u2005K; \u03bbmax (ethanol)/nm 503 (log \u220a 2.90) and 325(3.99); \u03bd (diamond anvil)/cm\u22121 2953br, 1688s, 1547vs, 1401vs, 1310vs, 1251vs, 1141vs, 871vs, 853s, 711vs, 654vs, 579vs, 454s and 420s; m/z (orbitrap ASAP) 139.0498 , C6H7N2O2 requires 139.0502. The UV/visible spectrum of (I)1,2,4,5-Benzene\u00adtetra\u00adamine tetra\u00adhydro\u00adchloride in distilled water (75\u2005ml) was treated with an excess of KUiso(H) = 1.2Ueq(carrier) was applied in all cases. The crystal studied was found to be a twin with the components related by a 180\u00b0 rotation about [001].Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016005107/sj5497sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016005107/sj5497Isup2.hklStructure factors: contains datablock(s) I. DOI: 1470620CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A 1:1.4 molar equivalent of benzene-1,3,5-tri\u00adcarb\u00adoxy\u00adlic acid cocrystallized with 4-hy\u00addroxy\u00adpyridine yields the 4-hy\u00addroxy\u00adpyridin-1-ium 3,5-di\u00adcarb\u00adoxy\u00adbenzoate salt. 5H6NO+\u00b7C9H5O6\u2212, (I), shows that 4-hy\u00addroxy\u00adpyridine has abstracted an H atom from benzene-1,3,5-tri\u00adcarb\u00adoxy\u00adlic acid, yielding a pyridinium cation and carboxyl\u00adate anion. The two ions form an extensive three-dimensional hydrogen-bonded network throughout the crystal. The hydrogen bonds that comprise the core of the network are considered strong, with O\u2014H\u22efO and N\u2014H\u22efO donor-to-acceptor distances ranging from 2.533\u2005(2) to 2.700\u2005(2)\u2005\u00c5. Packing is further enhanced by \u03c0-stacking of the cations and anions with like species [centroid\u2013centroid distance = 3.6206\u2005(13)\u2005\u00c5].The structure of the title salt, C This result allows for the hy\u00addroxy O and pyridine N atom to both act as hydrogen-bond donors, rather than the donor/acceptor situation of the 4-pyridone species. These two mol\u00adecules have been incorporated as linker species in metal\u2013organic frameworks shows that the 4-hy\u00addroxy\u00adpyridine has abstracted an H atom from the benzene\u00adtri\u00adcarb\u00adoxy\u00adlic acid, yielding a pyridinium cation and a carboxyl\u00adate anion and 1.348\u2005(3)\u2005\u00c5, respectively]. The remaining bonds within the ring display typical aromatic distances [C2\u2014C3 = 1.405\u2005(3)\u2005\u00c5 and C3\u2014C4 = 1.402\u2005(3)\u2005\u00c5]. The C3\u2014O1 distance of 1.326\u2005(2)\u2005\u00c5 is typical for a hy\u00addroxy O atom bound to an aromatic ring. Bond angles within the pyridine ring are unexceptional.The structure of \u2005\u00c5 and C14\u2014O7 = 1.332\u2005(3)\u2005\u00c5; C12\u2014O2 = 1.224\u2005(2)\u2005\u00c5 and C14\u2014O6 = 1.204\u2005(3)\u2005\u00c5]. The remaining carboxyl\u00adate group displays C\u2014O bond distances that are similar to each other and indicate delocalization of the C\u2014O bonds , supporting the proposed single negative charge on the benzene\u00adtri\u00adcarb\u00adoxy\u00adlic acid mol\u00adecule. This is further supported by the presence of H atoms, located in a difference Fourier map, on atoms O3 and O7. Bond distances and angles within the benzene ring are as expected.i [symmetry code: (i) \u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0\u2212\u00a0ii and O2iii, respectively [symmetry codes: (ii) \u2212x, \u2212y, z\u00a0\u2212\u00a0x, \u2212y\u00a0+\u00a01, z\u00a0+\u00a0The local inter\u00admolecular contacts consist of the pyridinium cation forming a hydrogen bond from the hy\u00addroxy group to the anionic carboxyl\u00adate group \u2005\u00c5, i.e. the c-axis spacing. The centroid-to-perpendicular distances are 3.3629\u2005(9)\u2005\u00c5 for the cation and 3.4372\u2005(9)\u2005\u00c5 for the anion. Both measurements are within accepted \u03c0\u2013\u03c0 contact ranges -one in MeOH (3\u2005ml) in a 20\u2005ml vial was added a solution of 4-hy\u00addroxy\u00adpyridine in MeOH (3\u2005ml). The mixture was shaken vigorously, covered with perforated Parafilm and allowed to evaporate slowly over a period of 5\u2005d, yielding colorless rod-like crystals.Uiso(H) = 1.2 Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a03x parameter refined to 0.20\u2005(8), which suggests the possibility of a small amount of inversion twinnning I, New_Global_Publ_Block. DOI: 10.1107/S2056989015011780/pk2555Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015011780/pk2555Isup3.cmlSupporting information file. DOI: 1407819CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Additional \u03c0\u2013\u03c0 inter\u00adactions between pyridinium rings stabilize this arrangement.In the crystal structure of the title compound, the mol\u00adecular components, 7H11N2)3[Cr(C2O4)3]\u00b74H2O, the central CrIII ion of the complex anion (point group symmetry 2) is coordinated by six O atoms from three chelating oxalate(2\u2212) ligands in a slightly distorted octa\u00adhedral coordination sphere. The Cr\u2014O bond lengths vary from 1.9577\u2005(11) to 1.9804\u2005(11)\u2005\u00c5, while the chelate O\u2014Cr\u2014O angles range from 82.11\u2005(6) to 93.41\u2005(5)\u00b0. The 4-(di\u00admethyl\u00adamino)\u00adpyridinium cations are protonated at the pyridine N atoms. In the crystal, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds link the cations and anions into a three-dimensional network. \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings of adjacent cations provide additional stabilization of the crystal packing, with the closest centroid-to-centroid distances being 3.541\u2005(1) and 3.575\u2005(1)\u2005\u00c5.In the title hybrid salt, (C The chelate O\u2014Cr\u2014O angles range from 82.11\u2005(6) to 93.41\u2005(5)\u00b0. The Cr\u2014O bond lengths vary from 1.9577\u2005(11) to 1.9804\u2005(11)\u2005\u00c5 and are similar to those found in the guanidinium tris\u00adchromate(III) salt \u00adpyridinium cations and lattice water mol\u00adecules Table\u00a01. In addi6(H2O)17[Cr(C2O4)3]4}\u00b77H2O , with an aqueous solution of 4-(di\u00admethyl\u00adamino)\u00adpyridine and oxalic acid . The mixture was stirred at 333\u2005K for about 30 minutes and then cooled to room temperature and filtered. The title compound crystallized by slow evaporation of the solvent at room temperature in form of light-violet crystals with dimensions up to 3\u2005mm within a few weeks.The title compound was obtained by reaction of an aqueous solution of the freshly prepared barium-oxalatochromate(III) salt {BaUiso(H) = 1.2Ueq(C) for aromatic and 0.96\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for methyl H atoms. H atoms of water mol\u00adecules as well as those bonded to N atoms were located from a difference Fourier map. Water H atoms were refined with soft restraints on O\u2014H and H\u22efH distances [O\u2014H = 0.82\u2005(1)\u2005\u00c5 and H\u22efH = 1.30\u2005(2)\u2005\u00c5] and Uiso(H) = 1.5Ueq(O) whereas H atoms bonded to N atoms were refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015020113/wm5230sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015020113/wm5230Isup2.hklStructure factors: contains datablock(s) I. DOI: 1400490CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports5: Article number: 17676; 10.1038/srep17676 Published online: 12032015; Updated: 04072016This Article contains an error in Supplementary Figure 1 where the value of the male chemotaxis index towards ascr#9 \u20180.26\u2009\u00b1\u20090.07\u2019 was incorrectly depicted as \u20180.38\u2009\u00b1\u20090.07\u2019. The correct Supplementary Figure 1 appears below as"} +{"text": "N-[2-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl]benzamides are reported. The 3-fluoro\u00adbenzamide crystallized with two independent mol\u00adecules in the asymmetric unit; the dihedral angles between the two benzene rings are 43.94\u2005(8) and 55.66\u2005(7)\u00b0. In the 3-bromo\u00adbenzamide and the 3-iodo\u00adbenzamide, this dihedral angle is much smaller, viz. 10.40\u2005(12) and 12.5\u2005(2)\u00b0, respectively.The crystal structures of three 14H9F4NO, (I), C14H9BrF3NO, (II), and C14H9F3INO, (III), the two benzene rings are inclined to one another by 43.94\u2005(8)\u00b0 in mol\u00adecule A and 55.66\u2005(7)\u00b0 in mol\u00adecule B of compound (I), which crystallizes with two independent mol\u00adecules in the asymmetric unit, but by only 10.40\u2005(12)\u00b0 in compound (II) and 12.5\u2005(2)\u00b0 in compound (III). In the crystals of all three compounds, N\u2014H\u22efO hydrogen bonds link the mol\u00adecules to form chains propagating along the a-axis direction for (I), and along the b-axis direction for (II) and (III). In the crystal of (I), \u2013A\u2013B\u2013A\u2013B\u2013 chains are linked by C\u2014H\u22efO hydrogen bonds, forming layers parallel to (010). Within the layers there are weak offset \u03c0\u2013\u03c0 inter\u00adactions present [inter\u00adcentroid distances = 3.868\u2005(1) and 3.855\u2005(1)\u2005\u00c5]. In the crystals of (II) and (III), the chains are linked via short halogen\u2013halogen contacts [Br\u22efBr = 3.6141\u2005(4)\u2005\u00c5 in (II) and I\u22efI = 3.7797\u2005(5)\u2005\u00c5 in (III)], resulting in the formation of ribbons propagating along the b-axis direction.In the title compounds, C The dihedral angle between the two benzene rings is 43.94\u2005(8)\u00b0 in mol\u00adecule A, while in mol\u00adecule B it is larger, being 55.66\u2005(7)\u00b0. The torsion angle of the central \u2013Car\u2014C(=O)\u2014N\u2014Car\u2013 segment is 176.74\u2005(12)\u00b0 in mol\u00adecule A and \u2212179.58\u2005(12)\u00b0 in mol\u00adecule B.The mol\u00adecular structure of compound (I)am Fig.\u00a02. In both3 substitution on the aniline ring are anti to one another, and the 3-bromo and 3-iodo substituents are anti to the N\u2014H bond in the central \u2013Car\u2014C(=O)\u2014N\u2014Car\u2013 segment of the mol\u00adecules, similar to situation observed in (I)A and B of compound (I)ar\u2014C(=O)\u2014N\u2014Car\u2013 segment is \u2212175.5\u2005(2)\u00b0 in (II)A and B of compound (I)The mol\u00adecular structures of compounds (II)A\u2013B\u2013A\u2013B\u2013 C(4) chains running along the a-axis direction \u2005\u00c5 and Cg2\u22efCg3i = 3.8553\u2005(9)\u2005\u00c5; Cg1 and Cg3 are the centroids of the aniline rings C1\u2013C6 and C15\u2013C20, respectively; Cg2 and Cg4 are the centroids of the benzoic acid rings C8\u2013C13 and C22\u2013C27, respectively; symmetry code (i) x\u00a0\u2212\u00a01, y, z]. The crystal structure does not feature any C\u2014H\u22efF or F\u22efF inter\u00adactions n Table\u00a01. Neighbos Table\u00a01, formingne Fig.\u00a06. Within ns Fig.\u00a06.C(4) chains running parallel to the b axis \u2005\u00c5], forming ribbons along [010]; see Fig.\u00a07The crystal structure of (II)is Fig.\u00a07. AdjacenC(4) chains running parallel to the b axis (Table\u00a03via short I\u22efI contacts [3.7797\u2005(5)\u2005\u00c5], forming ribbons along [010]; see Fig.\u00a08The crystal structure of (III)s Table\u00a03. Adjacenmeta position of the benzoic acid ring have a similar effect on the mol\u00adecular conformations and the supra\u00admolecular architectures exhibited by this class of compounds, whereas the fluoro substitution has a very different influence. For instance, there are two mol\u00adecules in the asymmetric unit of (I)A and B) of (I)From the above observations, it can be concluded that the bromo and iodo substitutions on the et al., 2016viz. N-(2-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl)aryl\u00adamides, gave four hits. They include N-(2-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl)benzamide, for which there are three reports: JOZFUB and JOZFUB01 in space group P43 phen\u00adyl)benzamide \u2014Car\u2013 segment is 175.1\u2005(5)\u00b0, which is very close to the values observed for the two independent mol\u00adecules in compound (I)meta position of the benzoyl ring, as in compound (I)A search of the Cambridge Structural Database were dissolved in phospho\u00adrous oxychloride taken in a 250\u2005ml round-bottomed flask. The mixtures were refluxed for an hour and later cooled to 273\u2005K. An equimolar amount of 2-(tri\u00adfluoro\u00admeth\u00adyl)aniline was added dropwise to these mixtures with continuous stirring. After completion of the addition, the reaction mixtures were brought to room temperature and stirring was continued for 1\u2005h. The reaction mixtures were poured into ice-cold water. The solids that separated were washed thoroughly with water, followed by washing with dilute hydro\u00adchloric acid, water, aqueous sodium hydrogen carbonate solution and again with water. The compounds were filtered under suction, dried and recrystallized from aqueous ethanol to constant melting points. Prismatic colourless single crystals of all three compounds were obtained by slow evaporation of solutions in methanol, with a few drops of water.Uiso = 1.2Ueq(C). In the final cycles of refinement of compound (III)R1, wR2, and GOF.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989016007866/su5298sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989016007866/su5298Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016007866/su5298IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989016007866/su5298IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989016007866/su5298Isup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016007866/su5298IIsup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016007866/su5298IIIsup7.cmlSupporting information file. DOI: 1479657, 1479656, 1479655CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal of the title compound, a supra\u00admolecular sheet structure is formed through N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. 19H19NO5, the amide carbonyl O atom is positioned anti to the other two carbonyl O atoms. The 4-hy\u00addroxy\u00adhydro\u00adcinnamate fragment is disordered over two positions with an occupancy ratio of 0.729\u2005(12):0.271\u2005(12). The N\u2014(C=O)\u2014C plane of the acetamide group and the acetate O\u2014(C=O)\u2014C plane are almost co-planar; the acetamide plane makes dihedral angles of 1.9\u2005(6) and 16.0\u2005(19)\u00b0, respectively, with the acetate planes of the major and minor occupancy components. In the crystal, N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a supra\u00admolecular sheet structure parallel to (102).In the title compound, C The carbonyl O1 and O16 atoms are positioned anti with respect to the carbonyl O12 atom. These C=O bond lengths are in the range 1.176\u2005(12)\u20131.226\u2005(6)\u2005\u00c5.The fragment O1/O12/N10/C2\u2013C9/C11/C13 including the acetamide group is almost planar with an r.m.s. deviation of 0.034\u2005(11)\u2005\u00c5. The 4-hy\u00addroxy\u00adhydro\u00adcinnamate fragment is disordered over two positions with occupancy ratio of 0.729\u2005(12):0.271\u2005(12). The acetamide plane O12/N10/C11/C12 makes dihedral angles of 1.9\u2005(6) and 16.0\u2005(19)\u00b0, respectively, with the disordered acetate planes O14/O16/C15/C17 and O14via N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds -2-chloro\u00adacetamide in the presence of dimethyl formamide (DMF) solvent and tri\u00adethyl\u00adamine base as eluent. The single crystals were obtained from a solvent mixture of ethyl acetate/n-hexane (3:1) upon slow evaporation at room temperature . FTIR \u03bdmax cm\u22121: 3428 (N\u2014H), 3354 (O\u2014H), 2971 (sp2 C\u2014H), 2887 (sp3 C\u2014H), 1735 (C=O ester), 1646 (C=O amide), 1601 (C=C aromatic), 1154 .The title compound was synthesized by direct condensation of 4-hy\u00addroxy\u00adphenyl propanoic acid with se Fig.\u00a04. The reaA, O25A and C15A\u2013C24A of the minor component were refined isotropically. Planarity restraints were applied for atoms C18\u2013C24, O25, C18A\u2013C24A and O25A. Bond-distance restraints were also applied for C20, C22, C23, O16A and C15A\u2013C24A. H10 and H25 of the NH and OH groups, respectively, were located in a difference Fourier map and the coordinates were refined with Uiso(H) = 1.2Ueq(N) and 1.5Ueq(O) [N\u2014H = 0.86\u2005(5)\u2005\u00c5 and O\u2014H = 0.95\u2005(12)\u2005\u00c5]. H25A of the minor occupancy OH group was refined with a restraint of O\u2014H = 0.90\u2005(2)\u2005\u00c5, and with Uiso(H) = 1.5Ueq(O). All other H atoms were included as riding atoms, with C\u2014H = 0.93\u20130.97\u2005\u00c5 and with Uiso(H) = 1.5Ueq(C) for methyl H atoms or 1.2Ueq(C) otherwise.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901600894X/is5453sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901600894X/is5453Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901600894X/is5453Isup3.cmlSupporting information file. DOI: 1483293CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II cation is chelated by four pyrrole-N atoms of the porphyrinate anion and coordinated by a pyridyl-N atom of the 4-cyano\u00adpyridine axial ligand in a distorted square-pyramidal geometry. The non-coordinating 4-cyano\u00adpyridine mol\u00adecule is disordered over two positions in the supra\u00admolecular channel formed by complex mol\u00adecules.In the crystal, the Zn 72H44N4O8)(C6H4N2)]\u00b7C6H4N2 or [Zn(TPBP)(4-CNpy]\u00b7(4-CNpy) , the ZnII cation is chelated by four pyrrole-N atoms of the porphyrinate anion and coordinated by a pyridyl-N atom of the 4-CNpy axial ligand in a distorted square-pyramidal geometry. The average Zn\u2014N(pyrrole) bond length is 2.060\u2005(6)\u2005\u00c5 and the Zn\u2014N(4-CNpy) bond length is 2.159\u2005(2)\u2005\u00c5. The zinc cation is displaced by 0.319\u2005(1)\u2005\u00c5 from the N4C20 mean plane of the porphyrinate anion toward the 4-cyano\u00adpyridine axial ligand. This porphyrinate macrocycle exhibits major saddle and moderate ruffling and doming deformations. In the crystal, the [Zn(TPBP)(4-CNpy)] complex mol\u00adecules are linked together via weak C\u2014H\u22efN, C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions, forming supra\u00admolecular channels parallel to the c axis. The non-coordinating 4-cyano\u00adpyridine mol\u00adecules are located in the channels and linked with the complex mol\u00adecules, via weak C\u2014H\u22efN inter\u00adactions and \u03c0-\u03c0 stacking or via weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. The non-coordinating 4-cyano\u00adpyridine mol\u00adecule is disordered over two positions with an occupancy ratio of 0.666\u2005(4):0.334\u2005(4).In the title compound, [Zn(C However, only three structures of zinc\u20134-NCpy non-porphyrinic species [CSD refcodes CYPYZN (4-CNpy)]\u00b7(4-CNpy) (I)During the last two decades, renewed attention to zinc metalloporphyrins has been noted for their applications in different fields II cation of the [Zn(TPBP)(4-CNpy)] complex has a distorted square-pyramidal coordination geometry bond length [2.159\u2005(2)\u2005\u00c5] is in the range (2.055\u20132.248\u2005\u00c5) of those of the zinc\u20134-CNpy complexes reported in the literature [CSD refcodes LIMWUZ ] [CSD refcodes ATUSOX . This value is close to those of the related five-coordinated zinc metalloporphyrins [Zn(TPP)(DMSO)] nearly bis\u00adects the \u2018cis\u2019 Np\u2014Fe\u2014Np angle, which is also the case for the title zinc\u20134-CNpy deriv\u00adative (I)The via weak non-classical C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and by C\u2014H\u22ef\u03c0 inter\u00adactions \u2005\u00c5 and 3.457\u2005(4)\u2005\u00c5, respectively.Within the crystal structure of (I)s Table\u00a01. The nitc axis atom C82A of the free 4-NCpy mol\u00adecule and atom O8 of a TPBP porphyrin [C82A\u2014H82A\u22efO8 distance = 3.226\u2005(5)\u2005\u00c5], (ii) the centroid (Cg18) of the C80A\u2013C81A\u2013C82A\u2013N8A\u2013C83A\u2013C84A ring of the disordered free 4-CNpy mol\u00adecule and the carbon atom C49 of an adjacent TPBR porphyrinato ligand with a C49\u2014H49\u22efCg18 contact length of 3.448\u2005(4)\u2005\u00c5, (iii) by aromatic \u03c0\u2013\u03c0 inter\u00adactions between the centroid (Cg19) of the C80A\u2013C81B\u2013C82B\u2013N8B\u2013C83B\u2013C84B ring of a free disordered 4-CNpy mol\u00adecule and the centroid (Cg11) of the phenyl porphyrin ring C28\u2013C33 and the N8B nitro\u00adgen atom of this second 4-CNpy mol\u00adecule is weakly bonded to the carbon atom C72 of a phenyl ring of a nearby TPBR porphyrinato ligand [C72__H72\u22efN8B distance = 3.226\u2005(15)\u2005\u00c5] (4-CNpy)] complexes parallel to the is Fig.\u00a04. Each fr\u00c5] Fig.\u00a05.2TPBP) and the starting [Zn(TPBP)] complex were synthesized using modified reported methods dissolved in 33\u2005mL of di\u00adchloro\u00admethane was added dropwise and stirred at 273\u2005K and then at room temperature for 12\u2005h. Upon completion, the reaction mixture was filtered and the solvent was evaporated to dryness, to afford 9.3\u2005g of a pale-yellow solid (yield 86%), m.p. = 356\u2013358\u2005K, C14H10O3: C 74.33, H 4.46%; found: C 73.98, H 4.35%. Spectroscopic analysis: 1H NMR \u03b4H (p.p.m.) 10.04 , 8.17 , 8.04 , 7.80 , 7.64 , 7.56 . 13C NMR \u03b4C (p.p.m.) 192.09, 164.12, 155.21, 134.31, 134, 131.13, 129.91, 129.03, 128.47, 122.90.Benzoic acid , 4-hy\u00addroxy\u00adbenzaldehyde and di\u00admethyl\u00adamino\u00adpyridin DMAP were dissolved at 273\u2005K in 20\u2005mL of di\u00adchloro\u00admethane. To this solution, 10.12\u2005g of v/v as an eluent). A purple solid was obtained and dried under vacuum .4.5\u2005mg of 4-formyl\u00adphenyl\u00adbenzoate (19.9\u2005mmol) were dissolved in 50\u2005mL of propionic acid. The solution was heated under reflex at 413\u2005K. Freshly distilled pyrrole was then added dropwise and the mixture was stirred for another 40\u2005min. The mixture was cooled overnight at 277\u2005K and filtered under vacuum. The crude product was purified using column chromatography \u03b4 (p.p.m.) 8.94 , 8.39 , 8.29 , 7.71 , 7.62 , \u22122.80 . UV/Vis (CHCl3): \u03bbmax 420 (512.7), 516 (16.7), 552 (7.4), 591 (4.8), 646 (4.0).Spectroscopic analysis: 2TPBP porphyrin and [Zn(OAc)2]\u00b72H2O in CHCl3 (30\u2005mL) and CH3OH (5\u2005mL) was stirred at room temperature overnight. The solvent was evaporated and a light-purple solid of the [Zn(TPBP)] complex was obtained .A mixture of the H1H NMR \u03b4(p.p.m. ) 9.04 , 8.40 , 8.30 , 7.85 , 7.64 , \u22122.80 . UV/Vis (CHCl3):\u03bbmax (10\u22123 \u220a) 425 (613.5), 554 (23.0), 596 (6.9).Spectroscopic analysis: To a solution of [Zn(TPBP)] in di\u00adchloro\u00admethane (5\u2005mL) was added an excess of 4-cyano\u00adpyridine . The reaction mixture was stirred at room temperature for 2\u2005h. Single crystals of the title complex were obtained by diffusion of hexa\u00adnes through the di\u00adchloro\u00admethane solution.1H NMR \u03b4(p.p.m. ) 9.04 , 8.40 , 8.30 , 7.67 , 7.53 . UV/Vis (CHCl3): \u03bbmax 425 (613.5), 554 (23.0), 596 (6.9).Spectroscopic analysis: \u22121 domain using a Perkin\u2013Elmer Spectrum Two FTIR spectrometer. The spectrum presents characteristic IR bands of the TPBP porphyrinato moiety. The C\u2014H stretching frequencies of the porphyrin are in the range 3060\u20132860\u2005cm\u22121, the ester group of the meso-substituents of this porphyrin are identified by a strong band at 1736\u2005cm\u22121, \u03bd(C=O) stretch and by two strong bands at 1264 and 1061 corresponding to the \u03bd(C\u2014O) stretching vibration. The IR spectrum of (I)\u22121 attributed to the nitrile stretching frequency \u03bd(C\u00a0N). The value of this band is almost identical to the one of the free 4-cyano\u00adpyridine (2236\u2005cm\u22121) which could be attributed both to the 4-CNpy ligand or the free 4-CNpy mol\u00adecule in (I)et al., 2000\u22121 (vw: very weak), 1523\u2005cm\u22121 (vw), 1505\u2005cm\u22121(w: weak), 1406\u2005cm\u22121 (m: medium), 996\u2005cm\u22121 (s: strong), 707\u2005cm\u22121 (m), 685\u2005cm\u22121 (m) and 538\u2005cm\u22121 (w) attributed to the pyridyl group of the coordinating and the free 4-cyano\u00adpyridine species (4-CNpy)]\u00b7(4-CNpy) (I)Uiso(H) = 1.2 Ueq(C). The non-coordinating 4-cyano\u00adpyridine mol\u00adecule is disordered over two positions A and B with refined occupancies of 0.666\u2005(4) and 0.334\u2005(4), respectively. The bond lengths and angles of this mol\u00adecule were restrained to ensure proper geometry using DFIX and DANG instructions of SHELXL2014 I, global. DOI: 10.1107/S2056989016000062/xu5882Isup2.hklStructure factors: contains datablock(s) I. DOI: 1445100CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The central C4N2O2 group of atoms in the dication are almost planar (r.m.s. deviation = 0.009\u2005\u00c5), and the carbonyl groups lie in an anti disposition to enable the formation of intra\u00admolecular amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds. To a first approximation, the pyridinium and amide N atoms lie to the same side of the mol\u00adecule [Npy\u2014C\u2014C\u2014Namide torsion angle = 34.8\u2005(2)\u00b0], and the anti pyridinium rings are approximately perpendicular to the central part of the mol\u00adecule [dihedral angle = 68.21\u2005(8)\u00b0]. In the anion, one carboxyl\u00adate group is almost coplanar with the ring to which it is connected [Cben\u2014Cben\u2014Cq\u2014O torsion angle = 2.0\u2005(3)\u00b0], whereas the other carboxyl\u00adate and carb\u00adoxy\u00adlic acid groups are twisted out of the plane . In the crystal, anions assemble into layers parallel to (10-4) via hy\u00addroxy-O\u2014H\u22efO(carbon\u00adyl) and charge-assisted hy\u00addroxy-O\u2014H\u22efO(carboxyl\u00adate) hydrogen bonds. The dications are linked into supra\u00admolecular tapes by amide-N\u2014H\u22efO(amide) hydrogen bonds, and thread through the voids in the anionic layers, being connected by charge-assisted pyridinium-N\u2014O(carboxyl\u00adate) hydrogen bonds, so that a three-dimensional architecture ensues. An analysis of the Hirshfeld surface points to the importance of O\u2014H\u22efO hydrogen bonding in the crystal structure.The asymmetric unit of the title salt, C N,N\u2032-bis\u00ad(pyridin-n-ylmeth\u00adyl)ethanedi\u00adamides, n = 2, 3 or 4, the mol\u00adecule with n = 2 appears to have attracted the least attention in co-crystallization studies; for the chemical structure of the diprotonated form of the n = 2 isomer see Scheme 1. By contrast, the n = 3 and 4 mol\u00adecules have attracted inter\u00adest from the crystal engineering community in terms of their ability to form co-crystals with iodo-containing species leading to aggregates featuring N\u22efI halogen bonding ethanedi\u00adamide conducted in ethanol. The harvested crystals were shown by crystallography to comprise (2-pyridinium)CH2N(H)C(=O)C(=O)CH2N(H)(2\u2013pyridinium) dications and 3,5-di\u00adcarb\u00adoxy\u00adbenzoate anions in the ratio 1:2; as the dication is located about a centre of inversion, one anion is found in the asymmetric unit. The confirmation for the transfer of protons during the co-crystallization experiment is found in (i) the pattern of hydrogen-bonding inter\u00adactions as discussed in Supra\u00admolecular features, and (ii) the geometric characteristics of the ions. Thus, the C\u2014N\u2014C angle in the pyridyl ring has expanded by over 3\u00b0 cf. that found in the only neutral form of N,N\u2032-bis\u00ad(pyridin-2-ylmeth\u00adyl)ethanedi\u00adamide characterized crystallographically in an all-organic mol\u00adecule, i.e. in a 1:2 co-crystal with 2-amino\u00adbenzoic acid and 1.250\u2005(2)\u2005\u00c5 is consistent with deprotonation and the formation of a carboxyl\u00adate group, and contrasts the great disparity in the C15\u2014O4, O5 [1.206\u2005(2) and 1.320\u2005(2)\u2005\u00c5] and C16\u2014O6, O7 [1.229\u2005(2) and 1.315\u2005(2)\u2005\u00c5] bond lengths.The title salt, Fig.\u00a014N2O2 chromophore is almost planar, having an r.m.s. deviation of 0.009\u2005\u00c5 and, from symmetry, the carbonyl groups are anti. An intra\u00admolecular amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bond is noted, Table\u00a02syn as seen in the value of the N1\u2014C1\u2014C6\u2014N2 torsion angle of 34.8\u2005(2)\u00b0. This planarity does not extend to the terminal pyridinium rings which are approximately perpendicular to and lying to either side of the central chromophore, forming dihedral angles of 68.21\u2005(8)\u00b0. The central C7\u2014C7i bond length of 1.538\u2005(4)\u2005\u00c5 is considered long for a C\u2014C bond involving sp2-hybridized atoms \u2005\u00c5, and almost perpendicular in the salt, with N1\u2014C1\u2014C6\u2014N2 being 73.84\u2005(15)\u00b0. These differences are highlighted in the overlay diagram shown in Fig.\u00a02In the dication, the central Cet al., 2009et al., 2015et al., 1998i.e. C_IV \u00b0, and for one of the carb\u00adoxy\u00adlic acid groups, i.e. 22.50\u2005(10)\u00b0.In the anion, the C13\u2014C8\u2014C14\u2014O2 and C9\u2014C10\u2014C15\u2014O4 torsion angles of 15.3\u2005(3) and 16.4\u2005(3)\u00b0, respectively, indicate twisted conformations between these residues and the ring to which they are attached whereas the C11\u2014C12\u2014C16\u2014O6 torsion angle of 2.0\u2005(3)\u00b0 shows this carb\u00adoxy\u00adlic acid group to be co-planar with the ring. The conformational flexibility in 3,5-di\u00adcarb\u00adoxy\u00adbenzoate anions is well illustrated in arguably the four most closely related structures in the crystallographic literature hydrogen bonds to form a familiar eight-membered {\u22efHOCO}2 synthon. These are connected by charge-assisted hy\u00addroxy-O\u2014H\u22efO(carboxyl\u00adate) hydrogen bonds that form C(8) chains. The result is a network of anions lying parallel to chains featuring pairs of amide-N\u2014H\u22efO(amide) hydrogen bonds and 10-membered {\u22efHNC2O}2 synthons, Fig.\u00a03b. The tapes are aligned along the a axis and, in essence, thread through the voids in the anionic layers to form a three-dimensional architecture, Fig.\u00a03c. The links between the anionic layers and cationic tapes are hydrogen bonds of the type charge-assisted pyridinium-N\u2014O(carboxyl\u00adate). In this scheme, no apparent role for the carbonyl-O4 atom is evident. However, this atoms accepts two C\u2014H\u22efO inter\u00adactions from pyridyl- and methyl\u00adene-H to consolidate the mol\u00adecular packing. Additional stabilization is afforded by pyridyl-C\u2014H\u22efO inter\u00adactions, Table\u00a02The mol\u00adecular packing may be conveniently described in terms of O\u2014H\u22efO hydrogen bonding to define an anionic network which is connected into a three-dimensional architecture by N\u2014H\u22efO hydrogen bonds; Table\u00a02Crystal Explorer 3.1 region between 1.7 and 2.7\u2005\u00c5. A small but significant contribution to the Hirshfeld surface of the dication due to N\u22efO/O\u22efN contacts is the result of inter\u00admolecular amide-N\u2014H\u22efO(amide) inter\u00adactions.The overall two-dimensional fingerprint plot (FP) of the salt together with those of the dication and anion, and FP\u2019s delineated into H\u22efH, O\u22efH/H\u22efO, C\u22efH/H\u22efC and C\u22efO/O\u22efC contacts are illustrated in Fig.\u00a07et al., 2014i.e. 0.8, due to a 23.7% contribution from the 54.5% available Hirshfeld surface and anti\u00adcipated 29.7% random contacts. The ER value of 1.4 corresponding to O\u22efH/H\u22efO contacts results from a relatively high 43.2% contribution by O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions. The carbon and oxygen atoms involved in the inter\u00admolecular C\u2014H\u22efO inter\u00adactions and short inter C\u22efO/O\u22efC contacts are at distances shorter than the sum of their respective van der Waals radii, hence they also have a high formation propensity, so the ER value is > 1. The C\u22efH/H\u22efC contacts in the crystal are enriched due to the poor nitro\u00adgen content and the presence of short inter\u00adatomic C\u22efH/H\u22efC contacts so the ratio is close to unity, i.e. 0.99. Finally, the ER value of 1.68 corresponding to N\u22efO/O\u22efN contacts for the surface of dication is the result of the charge-assisted N\u2014H\u22efO inter\u00adactions consistent with their high propensity to form.The inter\u00admolecular inter\u00adactions were further analysed using a recently reported descriptor, the enrichment ratio, ER ethanedi\u00adamide (LH2), has not been as well studied as the n = 3 and 4 isomers. This notwithstanding, the coordin\u00adation chemistry of LH2 is more advanced and diverse. Thus, co-crystals have been reported with a metal complex, i.e. [Mn3][ClO4]2\u00b7(LH2) 2 Pt}22+ \u00b7H2O}n 2(LH2)3}n chemistry, as in the aforementioned [CuL(LH2)(OH2)]2 , prepared in accord with the literature procedure (Schauer Uiso(H) set to 1.2Ueq(C). The oxygen- and nitro\u00adgen-bound H atoms were located in a difference Fourier map but were refined with distance restraints of O\u2014H = 0.84\u00b10.01\u2005\u00c5 and N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.5Ueq(O) and 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S2056989016000980/hb7560sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016000980/hb7560Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016000980/hb7560Isup3.cmlSupporting information file. DOI: 1447965CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Refinement of the site-occupancy factors of the three disordered bromide ions converges with occupancies 0.701\u2005(2), 0.831\u2005(2) and 0.456\u2005(2) summing to approximately two bromide ions per formula unit. The structure was refined as a two-component inversion twin with volume fractions 0.109\u2005(8):0.891\u2005(8) for the two domains. The central C3N unit of the bis\u00adamidinium ion is linked to the aliphatic propyl chain by a C\u2014N single bond. The other two bonds in this unit have double-bond character as have the four C\u2014N bonds to the outer NMe2 groups. In contrast, the three C\u2014N bonds to the central N atom of the (di\u00admethyl\u00adaza\u00adnium\u00adyl)propyl group have single-bond character. Delocalization of the two positive charges occurs in the N/C/N and C/N/C planes, while the third positive charge is localized on the di\u00admethyl\u00adammonium group. The crystal structure is stabilized by O\u2014H\u22efO, N\u2014H\u22efBr, O\u2014H\u22efBr and C\u2014H\u22efBr hydrogen bonds, forming a three-dimensional network.The asymmetric unit of the title hydrated salt, C N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adchloro\u00adformamidinium chloride, see: Tiritiris & Kantlehner (2008N-[3-(di\u00admethyl\u00adamino)\u00adprop\u00adyl]-N--N\u2032,N\u2032,N\u2032\u2032,N\u2032\u2032-tetra\u00admethyl\u00adguanidinium bis\u00ad(tetra\u00adphenyl\u00adborate), see: Tiritiris & Kantlehner (2015N\u2032\u2032-[3-(di\u00admethyl\u00adamino)\u00adprop\u00adyl]-N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adguanidine, see: Tiritiris & Kantlehner \u00c5b = 12.2932 (7) \u00c5c = 10.6633 (6) \u00c5\u03b2 = 97.454 (3)\u00b0V = 1190.39 (12) \u00c53Z = 2K\u03b1 radiationMo \u22121\u03bc = 3.40 mmT = 100 K0.41 \u00d7 0.29 \u00d7 0.25 mmBruker Kappa APEXII DUO diffractometerTmin = 0.334, Tmax = 0.481Absorption correction: multi-scan 6391 reflections with Rint = 0.033R[F2 > 2\u03c3(F2)] = 0.031wR(F2) = 0.069S = 0.997244 reflections265 parameters1 restraintH atoms treated by a mixture of independent and constrained refinementmax = 0.37 e \u00c5\u22123\u0394\u03c1min = \u22120.23 e \u00c5\u22123\u0394\u03c1Absolute structure: refined as an inversion twinAbsolute structure parameter: 0.109 (8)APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S2056989015024305/sj5490Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015024305/sj5490fig1.tif. DOI: The structure of the title compound with displacement ellipsoids at the 50% probability level. All carbon-bonded hydrogen atoms are omitted for the sake of clarity.Click here for additional data file.10.1107/S2056989015024305/sj5490fig2.tifac . DOI: ac view).N\u2014H\u22efBr, O\u2014H\u22efBr and O\u2014H\u22efO hydrogen bonds (black dashed lines) in the crystal structure of the title compound . The N\u2014H\u22efBr, O\u2014H\u22efBr, O\u2014H\u22efO and C\u2014H\u22efBr hydrogen bonds are depicted by black dashed lines.Mol\u00adecular packing of the title compound (1443022CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Vortioxetine, a new drug used to treat patients with major depressive disorder, has been crystallized as the free base and its methanol monosolvate. In both structures, the vortioxetine mol\u00adecules have similar conformations. 18H22N2S, (1), systematic name 1-{2-[sulfan\u00adyl]phen\u00adyl}piperazine, a new drug used to treat patients with major depressive disorder, has been crystallized as the free base and its methanol monosolvate, C18H22N2S\u00b7CH3OH, (2). In both structures, the vortioxetine mol\u00adecules have similar conformations: in (1), the dihedral angle between the aromatic rings is 80.04\u2005(16)\u00b0 and in (2) it is 84.94\u2005(13)\u00b0. The C\u2014S\u2014C bond angle in (1) is 102.76\u2005(14)\u00b0 and the corresponding angle in (2) is 103.41\u2005(11)\u00b0. The piperazine ring adopts a chair conformation with the exocyclic N\u2014C bond in a pseudo-equatorial orientation in both structures. No directional inter\u00adactions beyond normal van der Waals contacts could be identified in the crystal of (1), whereas in (2), the vortioxetine and methanol mol\u00adecules are linked by N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds, generating [001] chains.Vortioxetine, C Views of the asymmetric units of (1) and (2), with atom labelling, are presented in Figs. 1A\u22efO1i [symmetry code: (i) x, \u2212y\u00a0+\u00a0z\u00a0+\u00a0c-axis direction (Table\u00a01There are no hydrogen bonds or \u03c0\u2013\u03c0 stacking inter\u00adactions linking the mol\u00adecules in (1), while in (2) the presence of the additional methanol solvent mol\u00adecule results in the formation of zigzag chains mediated by alternating O1\u2014H1\u22efN2 and N2\u2014H2n Table\u00a01. a packiVortioxetine was supplied by Zhejiang Jingxin Pharmaceutical Co., Ltd. Crystals of (1) and (2) suitable for X-ray diffraction were recrystallized by slow evaporation from aceto\u00adnitrile and methanol\u2013water solutions, respectively, at room temperature over a few days.Uiso(H) = 1.2Ueq or 1.5Ueq(carrier atom).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015012256/hb7395sup1.cifCrystal structure: contains datablock(s) 1, 2, global. DOI: 10.1107/S2056989015012256/hb73951sup3.hklStructure factors: contains datablock(s) 1, 2. DOI: 10.1107/S2056989015012256/hb73952sup4.hklStructure factors: contains datablock(s) 2. DOI: Click here for additional data file.10.1107/S2056989015012256/hb73951sup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015012256/hb73952sup5.cmlSupporting information file. DOI: 1408949, 1408948CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A and B) in the asymmetric unit, the A and B mol\u00adecules are linked via pairs of N\u2014H\u22efS hydrogen bonds, forming dimers with an via pairs of C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming ribbons propagating along [100].In the crystal of the title compound, which crystallized with two independent mol\u00adecules in the asymmetric unit. Both mol\u00adecules have an L-shape but differ in the orientation of the benzyl ring with respect to the 3,4-di\u00admeth\u00adoxy\u00adbenzyl\u00adidine ring, this dihedral angle is 65.59\u2005(8)\u00b0 in mol\u00adecule A and 73.10\u2005(8)\u00b0 in mol\u00adecule B. In the crystal, the A and B mol\u00adecules are linked via pairs of N\u2014H\u22efS hydrogen bonds, forming dimers with an R22(8) ring motif. The dimers are linked via pairs of C\u2014H\u22efO hydrogen bonds, giving inversion dimers of dimers. These units are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming ribbons propagating in the [100] direction.The title compound, C S-benzyl\u00addithio\u00adcarbazate (SBDTC) whose derivatives have shown promising biological activities in the asymmetric unit. Both mol\u00adecules have an l-shape but differ in the orientation of the benzyl ring with respect to the 3,4-di\u00admeth\u00adoxy\u00adbenzyl\u00adidine ring, this dihedral angle being 65.59\u2005(8) \u00b0 in mol\u00adecule A and 73.10\u2005(8) \u00b0 in mol\u00adecule B are 1.331\u2005(2) and 1.282\u2005(2)\u2005\u00c5, respectively, in mol\u00adecule A, and 1.336\u2005(2) and 1.280\u2005(2)\u2005\u00c5, respectively, in mol\u00adecule B. The shorter length of the C\u2014N bond suggests the existence of a double bond which belongs to the imine group. Similarly, the shorter C\u2014S bond length relative to that of suggests that the former possesses double-bond character, indicating that the mol\u00adecule exists in its thione form in the solid state. The functional group identities proposed from these bond lengths are further supported by data obtained from the IR analysis reported below. Furthermore, the bond distances in the title compound are similar to those found for other carbodi\u00adthio\u00adate-derived Schiff bases \u2014C bond lengths (C1\u2014N1 and N2\u2014C9 in A and B) crystallizes in the conformer in which the two aromatic rings of the compound are cis with respect to each other across the C=N bonds, while the thione sulfur atom is trans with respect to the same bond.Both mol\u00adecules ring motif -2-benzyl\u00adidenehydrazine-1-carbodi\u00adthio\u00adates gave 13 hits. One of these concerns a structure very similar to the title compound, namely benzyl (E)-2-(4-meth\u00adoxy\u00adbenzyl\u00adidene)hydrazine-1-carbodi\u00adthio\u00adate \u00b0 in mol\u00adecule A and 73.10\u2005(8)\u00b0 in mol\u00adecule B of the title compound.A search of the Cambridge Structural Database . IR : 3360, 3122, 1602, 1069, 1023, 950, 788, 695. LCMS (ESI+): 347.1 [M+H]+.1.98\u2005g (0.01\u2005mol) of Uiso(H) = 1.5Ueq(C) for methyl H atoms and = 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901500095X/su5044sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901500095X/su5044Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901500095X/su5044Isup3.cmlSupporting information file. DOI: 1043887CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atom in this one-dimensional coordination polymer, with aqua, 4-formyl\u00adbenzoate and bridging pyrazine ligands, is located on a twofold rotation axis and has a slightly distorted octa\u00adhedral coordination sphere. Strong intra\u00admolecular O\u2014H\u22efO hydrogen bonds link the water mol\u00adecules to the carboxyl\u00adate O atoms.The Co 8H5O3)2(C4H4N2)(H2O)2]n, the CoII atom is located on a twofold rotation axis and has a slightly distorted octa\u00adhedral coordination sphere. In the equatorial plane, it is coordinated by two carboxyl\u00adate O atoms of two symmetry-related monodentate formyl\u00adbenzoate anions and by two N atoms of two bridging pyrazine ligands. The latter are bis\u00adected by the twofold rotation axis. The axial positions are occupied by two O atoms of the coordinating water mol\u00adecules. In the formyl\u00adbenzoate anion, the carboxyl\u00adate group is twisted away from the attached benzene ring by 7.50\u2005(8)\u00b0, while the benzene and pyrazine rings are oriented at a dihedral angle of 64.90\u2005(4)\u00b0. The pyrazine ligands bridge the CoII cations, forming linear chains running along the b-axis direction. Strong intra\u00admolecular O\u2014H\u22efO hydrogen bonds link the water mol\u00adecules to the carboxyl\u00adate O atoms. In the crystal, weak O\u2014Hwater\u22efOwater hydrogen bonds link adjacent chains into layers parallel to the bc plane. The layers are linked via C\u2014Hpyrazine\u22efOform\u00adyl hydrogen bonds, forming a three-dimensional network. There are also weak C\u2014H\u22ef\u03c0 inter\u00adactions present.In the title polymeric compound, [Co(C Atoms N1 and N2 of the pyrazine ligand and Co1 are located on a twofold rotation axis x, y\u00a0+\u00a01, z] is 7.1193\u2005(4)\u2005\u00c5.The asymmetric unit of the title compound contains a Cois Fig.\u00a01. The pyron Fig.\u00a02. The disIIO4N2 coordination sphere is composed of two carboxyl\u00adate O atoms of two symmetry-related monodentate formyl\u00adbenzoate anions and two N atoms of two bridging pyrazine ligands, which are bis\u00adected by the twofold rotation axis. The axial positions are occupied by two O atoms (O4 and O4i) of the coordinating water mol\u00adecules.The equatorial plane of the CoA (C2\u2013C7) is 7.50\u2005(8)\u00b0, while the benzene and pyrazine rings are oriented at a dihedral angle of 64.90\u2005(4)\u00b0.The near equality of the C1\u2014O1 [1.272\u2005(2)\u2005\u00c5] and C1\u2014O2 [1.245\u2005(2)\u2005\u00c5] bonds in the carboxyl\u00adate group indicates a delocalized bonding arrangement, rather than localized single and double bonds. The Co\u2014N bond length is 2.165\u2005(9)\u2005\u00c5, while the Co\u2014O bond lengths are 2.0551\u2005(9)\u2005\u00c5 (for benzoate oxygen) and 2.1491\u2005(11)\u2005\u00c5 (for water oxygen), close to standard values. The Co1 atom is displaced by 0.1034\u2005(2)\u2005\u00c5 from the mean plane of the carboxyl\u00adate group (O1/C1/O2). The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring water\u22efOwater hydrogen bonds were located in a difference Fourier map and were refined freely. The methine H atom was also located in a difference Fourier map and the C\u2014H distance restrained to 0.984\u2005(13)\u2005\u00c5. The aromatic C-bound H atoms were positioned geometrically with C\u2014H = 0.93\u2005\u00c5, and constrained to ride on their parent atoms, with Uiso(H) = 1.2Ueq(C).The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a024\u00b77H2O in H2O (25\u2005ml) and pyrazine in H2O (25\u2005ml) with sodium 4-formyl\u00adbenzoate in H2O (70\u2005ml) at room temperature. The mixture was filtered and set aside to crystallize at ambient temperature for one week, giving orange single crystals.The title compound was prepared by the reaction of CoSO10.1107/S205698901500403X/wm5129sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901500403X/wm5129Isup2.hklStructure factors: contains datablock(s) I. DOI: 1051344CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Compound (I) was the expected product and compound (II) was the oxidation product from air exposure.The title compounds C 17H14BrNO2, (I), and C17H15NO3, (II), were obtained from the reaction of 6-meth\u00adoxy-3,4-di\u00adhydro-2H-naphthalen-1-one and 2-bromo\u00adnicotinaldehyde in ethanol. Compound (I) was the expected product and compound (II) was the oxidation product from air exposure. In the crystal structure of compound (I), there are no short contacts or hydrogen bonds. The structure does display \u03c0\u2013\u03c0 inter\u00adactions between adjacent benzene rings and adjacent pyridyl rings. Compound (II) contains two independent mol\u00adecules, A and B, in the asymmetric unit; both are non-planar, the dihedral angles between the meth\u00adoxy\u00adbenzene and 1H-pyridin-2-one mean planes being 35.07\u2005(9)\u00b0 in A and 35.28\u2005(9)\u00b0in B. In each mol\u00adecule, the 1H-pyridin-2-one unit participates in inter\u00admolecular N\u2014H\u22efO hydrogen bonding to another mol\u00adecule of the same type (A to A or B to B). The structure also displays \u03c0\u2013\u03c0 inter\u00adactions between the pyridyl and the benzene rings of non-equivalent mol\u00adecules .The title compounds C Our research strategy to synthesize novel compounds considered analogs of the natural product chalcone, which contains two aromatic rings and an \u03b1-\u03b2-unsaturated ketone. Chalcones, bioactive defense mol\u00adecules found in plants and used in traditional Chinese medicine, have demonstrated anti\u00adcancer, anti\u00adbacterial, anti\u00adfungal, and anti-inflammatory properties and O1B\u2014C1B = 1.257\u2005(3)\u2005\u00c5. The mol\u00adecules are also non-planar, the benzene\u2013pyridyl angle being 36.18\u2005(10)\u00b0 in A and 35.91\u2005(10)\u00b0 in B.Compound (I)Cg1i distance is 3.635\u2005(3)\u2005\u00c5 and has a \u2018face-on\u2019 geometry. There are two \u03c0\u2013\u03c0 inter\u00adactions in the crystal between adjacent benzene rings, Cg1\u22efCg1ii = 3.944\u2005(4)\u2005\u00c5 and between adjacent pyridyl rings, Cg2\u22efCg2iii = 3.639\u2005(4)\u2005\u00c5 . The \u03c0\u2013\u03c0 inter\u00adactions form ribbons in the ne Fig.\u00a03, which ane Fig.\u00a03.H-pyridin-2-one unit participates in inter\u00admolecular N\u2014H\u22efO hydrogen bonding, with a classical A to A or B to B), see Fig.\u00a05A\u2013B or B\u2013A inter\u00adactions) and Cg5\u22efCg6ii = 3.857\u2005(4)\u2005\u00c5 .In each one of the independent mol\u00adecules in (II)s) Fig.\u00a06; Cg3\u22efCg4et al., 2016et al., 2002et al., 2005et al., 2002A search of the Cambridge Structural Database and 2-bromo\u00adnicotinaldehyde (1\u2005mmol) were dissolved in ethanol (5\u2005mL). An NaOH solution was added and the reaction was stirred until a precipitate formed. The reaction mixture was cooled in an ice bath for 20 minutes. The solids were filtered off and recrystallized from MeOH/H2O. Slow evaporation of a methano\u00adlic solution gave dark purple/brown crystals, which proved to be 3-[(E)-meth\u00adyl]pyridin-2(1H)-one, (II), and lighter purple crystals which proved to be (E)-2-[(2-bromopyridin-3-yl)methyl\u00adidene]-6-meth\u00adoxy-3,4-di\u00adhydro\u00adnaph\u00adthalen-1(2H)-one, (I).6-Meth\u00adoxy-3,4-di\u00adhydro-2Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016009300/bg2587sup1.cifCrystal structure: contains datablock(s) General, I, II. DOI: 10.1107/S2056989016009300/bg2587Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016009300/bg2587IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1484124, 1484123CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In this model CD8+ T cells mediate biliary ductular damage, whereas CD4+ T cells mediate induction of colon-specific autoimmunity. Importantly, IL-2R\u03b1\u2212/\u2212 mice have high levels of interferon \u03b3 (IFN-\u03b3), and interleukin-17A (IL-17A). We produced unique double deletions of mice that were either IL-17A\u2212/\u2212IL-2R\u03b1\u2212/\u2212 or IFN-\u03b3\u2212/\u2212IL-2R\u03b1\u2212/\u2212 to specifically address the precise role of these two cytokines in the natural history of autoimmune cholangitis and colitis. Of note, deletion of IL-17A in IL-2R\u03b1\u2212/\u2212 mice led to more severe liver inflammation, but ameliorated colitis. In contrast, there were no significant changes in the immunopathology of double knock-out IFN-\u03b3\u2212/\u2212 IL-2R\u03b1\u2212/\u2212 mice, compared to single knock-out IL-2R\u03b1\u2212/\u2212 mice with respect to cholangitis or colitis. Furthermore, there was a significant increase in pathogenetic CD8+ T cells in the liver of IL-17A\u2212/\u2212IL-2R\u03b1\u2212/\u2212 mice. Our data suggest that while IL-17A plays a protective role in autoimmune cholangitis, it has a pro-inflammatory role in inflammatory bowel disease. These data take on particular significance in the potential use of anti-IL-17A therapy in humans with primary biliary cirrhosis.IFN-\u03b3 is a signature Th1 cell associated cytokine critical for the inflammatory response in autoimmunity with both pro-inflammatory and potentially protective functions. IL-17A is the hallmark of T helper 17 (Th17) cell subsets, produced by \u03b3\u03b4T, CD8+ T, NK and NKT cells. We have taken advantage of our colony of IL-2R\u03b1 We report herein that deletion of IL-17A in IL-2R\u03b1\u2212/\u2212 mice aggravated cholangitis but ameliorated colitis. In contrast, there was no significant effect of the deletion of IFN-\u03b3 on the immunopathology of either autoimmune cholangitis or colitis. Importantly, T cells, particularly CD8+ T cells, were significantly increased in IL-17A\u2212/\u2212IL-2R\u03b1\u2212/\u2212 mice. Our data suggests that IL-17A plays a protective role in autoimmune cholangitis but a proinflammatory role in colitis in IL-2R\u03b1\u2212/\u2212 mice.To examine in specific detail the role of IFN-\u03b3 versus IL-17A in autoimmune cholangitis and colitis, we took advantage of IL-17A\u2212/\u2212 and IFN-\u03b3\u2212/\u2212 (B6.129S7-IFN-\u03b3tm1Ts) mice on a C57B/6J background were initially obtained from the Jackson Laboratory . IL-17A\u2212/\u2212 mice were donated by Dr. Yoichiro Iwakura . All genetically modified mice studied herein have been backcrossed to C57BL/6 background for at least 10 generations. IL-2R\u03b1\u2212/\u2212 were bred to heterozygosity to these strains. To generate IL-17A\u2212/\u2212IL-2R\u03b1\u2212/\u2212 and IFN-\u03b3\u2212/\u2212IL-2R\u03b1\u2212/\u2212 mice, IL-17A\u2212/\u2212 or IFN-\u03b3\u2212/\u2212 mice were mated with IL-2R\u03b1+/\u2212 mice to obtain IL-17A+/\u2212IL-2R\u03b1+/\u2212 or IFN-\u03b3+/\u2212IL-2R\u03b1+/\u2212 mice, which were subsequently backcrossed with IL-17A\u2212/\u2212 or IFN-\u03b3\u2212/\u2212 mice to obtain IL-17A\u2212/\u2212IL-2R\u03b1+/\u2212 or IFN-\u03b3\u2212/\u2212IL-2R\u03b1+/\u2212 mice; IL-17A\u2212/\u2212IL-2R\u03b1\u2212/\u2212 or IFN-\u03b3\u2212/\u2212IL-2R\u03b1\u2212/\u2212 mice were obtained by inter-breeding. The IL-2R\u03b1 gene was identified by flow cytometric analysis based on mean fluorescent intensity of CD25. All mice were studied between 12 and 16 weeks of age and animals were individually housed in ventilated cages under the specific pathogen-free conditions. All animal experimental protocols were approved by the Institutional Animal Care and Use Committee of the School of Life Sciences, University of Science and Technology of China, Hefei, China. Animals were anesthetized for blood collection with Xylazine (5 mg/kg)/Ketamine (25 mg/kg) administered subcutaneously. Euthanasia was performed by an overdose of CO2 by inhalation, consistent with the recommendations on the Panel on Euthanasia of the American Veterinary Medical Association. There was no surgery performed and any animals exhibiting discomfort or distress by the veterinary staff were euthanized.IL-2R\u03b12. Cells were then stained for surface markers using anti-CD4 PerCP/CY5.5, anti-NK1.1 PE/CY7, anti-CD3 Pacific Blue and anti-CD8\u03b2 FITC (Biolegend), then intracellular stained with anti-IFN-\u03b3 PE and anti-IL-17A APC (Biolegend) after fixation and permeabilization with Fixation Buffer and Permeabilization Wash Buffer (Biolegend), Rat IgG1, \u03ba PE and Rat IgG1, \u03ba APC were used as isotype controls for anti-IFN-\u03b3 PE and anti-IL-17A APC respectively. Finally, samples were subjected to multiple-color analysis by BD FACSVerse flow cytometer (BD Biosciences). Acquired data were analyzed with FlowJo Software .Lymphocytes from spleen and liver were isolated using 40%/70% percoll Liver and colon were fixed in 4% paraformaldehyde, embedded in paraffin and cut into 4 and 6 \u00b5m sections respectively, then deparaffinized, stained with hematoxylin and eosin (H&E), and evaluated using light microscopy. The quantitation of tissue damage was performed by a \u201cblinded\u201d pathologist. Firstly, the degree of portal inflammation was evaluated and scored according to the most severe lesions as follows: 0, no change; 1, minimal inflammation; 2, mild inflammation; 3, moderate inflammation; 4, severe inflammation. In addition, the degree of inflammatory frequency in a specimen was determined by the percentage of affected tissue within the total hepatic lobules per specimen and coded as follows: 0, none, 1, 1%\u201310%; 2, 11\u201320%; 3, 21\u201350%; 4, more than 50%. Finally, a summary score that includes severity and frequency analysis was generated as the sum of these scores. We note that lobular inflammation was scored in the same fashion as portal inflammation. Second, bile duct damage was evaluated firstly by the degree of severity in the most severe lesions as follows: 0, no change; 1, epithelial damage (only cytoplasmic change); 2, epithelial damage with cytoplasmic and nuclear change; 3, non-suppurative destructive cholangitis (CNSDC); 4, bile duct loss. The frequency of bile duct damage was then scored as follows: 0, none; 1, 1%\u201310%; 2, 11\u201320%; 3, 21\u201350%; 4, more than 50%. To evaluate bile duct loss, bile duct epithelium was highlighted by immunostaining using rabbit anti-keratin wide antibody . To obtain an integrative evaluation, the scores of severity and frequency were added together. Third, colon histopathology was scored as follows: 0, no significant changes; 1, minimal scattered mucosal inflammatory cell infiltrates, with or without minimal epithelial hyperplasia; 2, mild scattered to diffuse inflammatory cell infiltrates, sometimes extending into the submucosa and associated with erosions, with mild to moderate epithelial hyperplasia and mild to moderate mucin depletion from goblet cells; 3, moderate inflammatory cell infiltrates that were sometimes transmural, with moderate to severe epithelial hyperplasia and mucin depletion; and 4, marked inflammatory cell infiltrates that were often transmural and associated with crypt abscesses and occasional ulceration, with marked epithelial hyperplasia, mucin depletion, and loss of intestinal glands \u2212\u0394\u0394Ct method Total RNA from liver and colon was extracted with RNeasy Mini Kit , and cDNA synthesized with the PrimeScript RT reagent Kit . Quantitative PCR was performed using a SYBR Premix Ex TaqTM II . Data were collected by an ABI StepOne real-time PCR system . The PCR primers used in this study are listed in The levels of IFN-\u03b3, tumor necrosis factor \u03b1 (TNF-\u03b1), IL-2, IL-4, IL-6 and IL-10 from serum were measured simultaneously with a cytometric bead array kit , using a FACSVerse flow cytometer with CBA software (BD Biosciences).Data are presented as the mean \u00b1 standard error. The histological scores of liver inflammation and colitis were compared using one-way analysis of variance (ANOVA) followed by Kruskal-Wallis multiple comparisons. The two-tailed unpaired Mann-Whitney test was used in other comparisons. A value of p<0.05 was considered statistically significant; a value of 0.05 4.7\u2005\u00c5).In the extended structure of (I)s Table\u00a01 to the sC(7) O2\u2014H2A\u22efO4i hydrogen bonds (Table\u00a02i to not form an inter\u00admolecular hydrogen bond (H3\u22efO4i = 3.2\u2005\u00c5), but instead forms an intra\u00admolecular link to O1. A very long inter\u00admolecular C\u2014H\u22efN inter\u00adaction is observed but there is no \u03c0\u2013\u03c0 stacking in (II)The extended structure of (II)s Table\u00a02 with alm1 screw axis, so that the C1-benzene ring is \u2018flipped\u2019 from one side of the chain to the other in adjacent mol\u00adecules. As noted above, the hydroxyl group is disordered over two orientations. The hydrogen bond from the major orientation of O2A\u2014H2A is still a bond to O4i forms an O\u2014H\u22efO hydrogen bond in the opposite chain direction to O1ii : O1 also accepts an intra\u00admolecular N\u2014H\u22efO hydrogen bond, as seen in (II)The packing in the centrosymmetric structure of (III)i Table\u00a03, where i2OH)CON(CH3)N=CH\u2013 fragments reported in Version 5.36 of the Cambridge Structural Database CONHN=CH\u2013 groupings with different substituents at each end of the fragment, all of which have been reported by us in the last few years -(S)-ROCONHCH(CH2OH)CONHN=CH-benzene compound was added to a solution of the appropriate , b = 13.003\u2005(19), c = 22.94\u2005(3)\u2005\u00c5, \u03b1 = 92.93\u2005(2), \u03b2 = 91.48\u2005(3), \u03b3 = 98.13\u2005(3)\u00b0, V = 2804\u2005(7)\u2005\u00c53]. An atomic model could be developed in space group P1 with Z = 6, but a PLATON ]. The C-bound H atoms were placed geometrically (C\u2014H = 0.95\u20131.00\u2005\u00c5) and refined as riding atoms. The constraint Uiso(H) = 1.2Ueq(carrier) or 1.5Ueq(methyl carrier) was applied in all cases. The H atoms of the hydroxyl groups were allowed to rotate about their C\u2014O bond (SHELXL HFIX 83 instruction with O\u2014H = 0.84\u2005\u00c5 and C\u2014O\u2014H = 109.5\u00b0) to best fit the electron density. The methyl groups were allowed to rotate, but not to tip, to best fit the electron density (AFIX 137 instruction).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989015010440/hg5443sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989015010440/hg5443Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989015010440/hg5443IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989015010440/hg5443IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989015010440/hg5443Isup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015010440/hg5443IIsup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015010440/hg5443IIIsup7.cmlSupporting information file. DOI: 1404006, 1404005, 1404004CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "These represent the chiral atropoisomers distinguished by the mutual arrangement of the two acet\u00adyl\u2013hydrazone groups with a cis conformation of the C=N bonds. The two cyclo\u00adpenta\u00addienyl (Cp) rings are planar and nearly parallel, the tilt between the two rings being 3.16\u2005(16)\u00b0 [4.40\u2005(18)\u00b0 for the second independent mol\u00adecule]. The conformation of the Cp rings is close to eclipsed, the twist angle being 0.1\u2005(2)\u00b0 [3.3\u2005(2)\u00b0]. The two acet\u00adyl\u2013hydrazone substituents are also planar and are inclined at 13.99\u2005(15)/9.17\u2005(16)\u00b0 [6.83\u2005(17)/14.59\u2005(15)\u00b0] relative to the Cp rings. The Fe\u2014C bond lengths range from 2.035\u2005(3) to 2.065\u2005(2)\u2005\u00c5, with an average of 2.050\u2005(3)\u2005\u00c5 , which agrees well with those reported for most ferrocene derivatives. In the crystal, the mol\u00adecules form dimers via two strong N\u2014H\u22efN hydrogen bonds. The dimers are linked into a three-dimensional framework by weak N\u2014H\u22efN hydrogen bonds.The title compound, [Fe(C DOI: 10.1107/S1600536814014366/rk2429Isup2.hklStructure factors: contains datablock(s) I. DOI: 1009066CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The CoII atoms are linked by the oxalate anions into a chain running parallel to [100]. The chains are linked by the BPMO ligands into a three-dimensional architecture. In addition, N\u2014H\u22efO hydrogen bonds stabilize the crystal packing.In the polymeric title compound, [Co(C DOI: 10.1107/S1600536814015608/bt6986Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814015608/bt6986fig1.tifx y z x y z x y z x y z . DOI: x\u00a0+\u00a01, \u2212y, \u2212 z\u00a0+\u00a01; (ii) \u2212x, \u2212y, \u2212 z\u00a0+\u00a01; (iii) x, \u2212 y\u00a0+\u00a0z\u00a0+\u00a0x\u00a0+\u00a01, \u2212 y\u00a0+\u00a0z\u00a0\u2212\u00a0A view of the mol\u00adecule of (I). Displacement ellipsoids are drawn at the 30% probability level. (i) \u2212 Click here for additional data file.10.1107/S1600536814015608/bt6986fig2.tif. DOI: View of the three-dimensional structure of (I).1012047CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II cations that are linked by thio\u00adcyanato anions into chains which are further connected into layers by inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efS hydrogen bonding via additional methanol mol\u00adecules.The crystal structure of the title compound consists of Cd 2(C6H6N2S)]\u00b72CH3OH}n, consists of one cadmium(II) cation that is located on a centre of inversion as well as one thio\u00adcyanate anion, one pyridine-4-carbo\u00adthio\u00adamide ligand and one methanol mol\u00adecule in general positions. The CdII cations are octa\u00adhedrally coordinated by the pyridine N atom of two pyridine-4-carbo\u00adthio\u00adamide ligands and by the S and N atoms of four thio\u00adcyanate anions and are linked into chains along [010] by pairs of anionic ligands. These chains are further linked into layers extending along (201) by inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efS hydrogen bonds. One of the amino H atoms of the pyridine-4-carbo\u00adthio\u00adamide ligand is hydrogen-bonded to the O atom of a methanol mol\u00adecule, and a symmetry-related methanol mol\u00adecule is the donor group to the S atom of another pyridine-4-carbo\u00adthio\u00adamide ligand whereby each of the pyridine-4-carbo\u00adthio\u00adamide ligands forms two pairs of centrosymmetric N\u2014H\u22efS and O\u2014H\u22efS hydrogen bonds. The methanol mol\u00adecules are equally disordered over two orientations.The asymmetric unit of the polymeric title compound, {[Cd(NCS) The CdII cation is sixfold coordinated by two N-bonding pyridine\u00adthio\u00adamide ligands as well as two N- and two S-coordinating thio\u00adcyanate anions in an all trans distorted octa\u00adhedral environment nt Fig.\u00a01. As expent Fig.\u00a01. The CdI0] Fig.\u00a02. The metvia the methanol solvent mol\u00adecules by inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efS hydrogen bonding es Fig.\u00a03. Each pyes Fig.\u00a03. The hydes Fig.\u00a03. This aret al., 2014According to the Cambridge Structural Database 2\u00b73H2O were purchased from Alfa Aesar. Cd(NCS)2 was synthesized by stirring 17.5\u2005g (57.0\u2005mmol) Ba(NCS)2\u00b73H2O and 14.6\u2005g (57.0\u2005mmol) CdSO4\u00b73/8H2O in 300\u2005ml water at room temperature for 3\u2005h. The white residue of BaSO4 was filtered off and the resulting solution dried at 353\u2005K. The homogeneity of the product was checked by X-ray powder diffraction and elemental analysis. The title compound was obtained by reaction of 11.4\u2005mg Cd(NCS)2 (0.05\u2005mmol) with 27.6\u2005mg pyridine-4-carbo\u00adthio\u00adamide (0.2\u2005mmol) in boiling methanol (2\u2005ml). Crystals suitable for single-crystal x-ray diffraction formed after cooling.CdSOUiso(H) = 1.2Ueq (1.5 for methyl and O\u2014H hydrogen atoms) using a riding model with C\u2014H = 0.95\u2005\u00c5 for aromatic, C\u2014H = 0.98\u2005\u00c5 for methyl, N\u2014H = 0.88\u2005\u00c5 and O\u2014H = 0.84\u2005\u00c5, respectively. The methanol mol\u00adecule is equally disordered over two orientations and was refined using a split model using SAME restraints I, New_Global_Publ_Block. DOI: 10.1107/S2056989016002632/wm5271Isup2.hklStructure factors: contains datablock(s) I. DOI: 1453442CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S-benzyl-\u03b2-N-(2-hy\u00addroxy\u00adphenyl\u00adethyl\u00adidene)di\u00adthio\u00adcarbazate groups, connected through an S\u2014S single bond. The two moieties are twisted with a dihedral angle of 87.88\u2005(4)\u00b0 between the S2C=N planes.The title mol\u00adecule consists of two Schiff base moieties, namely two 32H30N4O2S4, consists of two Schiff base moieties, namely two S-benzyl-\u03b2-N-(2-hy\u00addroxy\u00adphenyl\u00adethyl\u00adidene)di\u00adthio\u00adcarbazate groups, which are connected through an S\u2014S single bond. These two moieties are twisted with respect to each other, with a dihedral angle of 87.88\u2005(4)\u00b0 between the S2C=N planes. A bifurcated intra\u00admolecular O\u2014H\u22ef hydrogen bond is observed in each moiety. In the crystal, mol\u00adecules are linked by pairs of C\u2014H\u22efO hydrogen bonds into inversion dimers. The dimers are further stacked in a column along the b axis through weak C\u2014H\u22ef\u03c0 inter\u00adactions.The title compound, C S-benzyl\u00addithio\u00adcarbazate (SBDTC) . The (imino\u00adeth\u00adyl)phenol fragments (C1\u2013C8/O1/N1 and C25\u2013C32/O2/N4) are essentially planar with maximum deviations of 0.0559\u2005(12)\u2005\u00c5 for N1 and 0.0200\u2005(11)\u2005\u00c5 for N4 and make dihedral angles of 18.17\u2005(4) and 17.49\u2005(4)\u00b0 with the N2/S1/S2/C9 and N3/S3/S4/C17 planes, respectively. The C\u2014S distances of 1.7461\u2005(12)\u20131.8220\u2005(13)\u2005\u00c5 are comparable to the values for the most similar di\u00adthio\u00adcarbazate derivatives di\u00adthio\u00adcarbazate was prepared by Pramanik et al. , was prepared according to the literature method . The prepared compound (0.17\u2005g) was dissolved in aceto\u00adnitrile (20\u2005ml) on warming and mixed with ethanol (10\u2005ml). Light-yellow platelet single crystals of the title compound (m.p. 386\u2013387\u2005K) suitable for X-ray study were obtained after 17 days along with colorless needle-shaped crystalline solids (m.p. 413\u2013418\u2005K).The ligand precursor, Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016002371/is5440sup1.cifCrystal structure: contains datablock(s) General, I. DOI: 10.1107/S2056989016002371/is5440Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016002371/is5440Isup3.cmlSupporting information file. DOI: 1452193CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "R stereochemical configuration. No head-to-tail hydrogen bonding is observed in the crystal packing, as is the case in estrone and other derivatives.A new ferrocene complex, 16-ferrocenylmethyl-3\u03b2-hy\u00addroxy\u00adestra-1,3,5(10)-trien-17-one, has been synthesized and structurally characterized by single-crystal X-ray diffraction techniques. The ferrocenylmethyl group is positioned at the \u03b2 face of the estrone moiety; as a result, a new stereogenic center is formed leading to an 5H5)(C24H27O2)]\u00b7C2H6OS, has been synthesized and structurally characterized by single-crystal X-ray diffraction techniques. The mol\u00adecule crystallizes in the space group P21 with one mol\u00adecule of dimethyl sulfoxide. A hydrogen bond links the phenol group and the dimethyl sulfoxide O atom, with an O\u22efO distance of 2.655\u2005(5)\u2005\u00c5. The ferrocene group is positioned in the \u03b2 face of the estrone moiety, with an O\u2014C\u2014C\u2014C torsion angle of 44.1\u2005(5)\u00b0, and the carbonyl bond length of the hormone moiety is 1.216\u2005(5)\u2005\u00c5, typical of a C=O double bond. The average Fe\u2014C bond length of the substituted Cp ring [Fe\u2014C(Cp*)] is similar to that of the unsubstituted one [Fe\u2014C(Cp)], i.e. 2.048\u2005(3) versus 2.040\u2005(12)\u2005\u00c5. The structure of the complex is compared with those of estrone and eth\u00adoxy\u00admethyl\u00adestrone.A new ferrocene complex, 16-ferrocenylmethyl-3\u03b2-hy\u00addroxy\u00adestra-1,3,5(10)-trien-17-one dimethyl sulfoxide monosolvate, [Fe(C In this context, we present herein the synthesis and crystal structure of 16-ferrocenylmethyl-3\u03b2-hy\u00addroxy\u00adestra-1,3,5(10)-trien-17-one dimethyl sulfoxide monosolvate (2) and compare it with the structure of estrogen (1) and 16\u03b2-eth\u00adoxy\u00admethyl\u00adestrone (3) \u2005\u00c5, which is very similar to in estrogen and 16\u03b2-eth\u00adoxy\u00admethyl\u00adestrone , corresponding to a carbon\u2013oxygen double (C=O) bond. However, the substitution at C16 of the steroid in 2 and 3, ferrocenylmethyl and eth\u00adoxy\u00admethyl groups, respectively, makes torsion angles and bond angles at the 16-position slightly different. Both substituents are located on the beta face but, the torsion angle (between C19 and carbonyl group) defined as C19\u2013C16\u2014C17\u2014O2 in 2 is smaller than in 3 (between the carbonyl and the meth\u00adoxy groups), 44.1\u2005(5) and 49.7\u2005(2)\u00b0, respectively. The ferrocene moiety is positioned at 112.6\u2005(3)\u00b0 from C16 (\u2220C20\u2014C19\u2014C16) while the eth\u00adoxy\u00admethyl group is at 108.4\u2005(1)\u00b0 (\u2220C16\u2014C1\u2014O3). The average Fe\u2014C bond length of the substituted Cp ring [Fe\u2014C vs 2.040\u2005(12)\u2005\u00c5 as mobile phase, affording 67% of 2 as a yellow solid. Yellow rod-shaped crystals were obtained after dissolving the solid 16-ferro\u00adcenyl\u00admethyl-3\u03b2-hy\u00addroxy\u00adestra-1,3,5(10)-trien-17-one in a solution of CH2Cl2 with a few drops of dimethyl sulfoxide, to assure a concentrate solution, layered in hexane.In a 500\u2005mL Parr bottle, 16-ferrocenyl\u00adidene-3\u03b2-hy\u00addroxy\u00adestra-1,3,5(10)-trien-17-one complex was dissolved in a mixture of tetra\u00adhydro\u00adfuran (THF) and ethanol (1:1) and Pd/C . The system was purged three times with Hd(C\u2014H) = 0.95\u2005\u00c5, Uiso(H) = 1.2Ueq(C); d(C\u2014H2) = 0.99\u2005\u00c5,Uiso(H) = 1.2 Ueq (C); d(C\u2014H3) = 0.98\u2005\u00c5, Uiso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016008446/bg2586sup1.cifCrystal structure: contains datablock(s) I. DOI: 1479699CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The 5-fluoro\u00adcytosine and melamine mol\u00adecules inter\u00adact via N\u2014H\u22efO, N\u2014H\u22efN and N\u2014H\u22efO, N\u2014H\u22efN, C\u2014H\u22efF hydrogen bonds.The asymmetric unit of the title compound comprises two independent 5-fluoro\u00adcytosine mol\u00adecules and one half-mol\u00adecule of melamine. The 5-fluoro\u00adcytosine mol\u00adecules are linked through two different homosynthons; one is formed 4H4FN3O\u00b7C3H6N6, comprises of two independent 5-fluoro\u00adcytosine (5FC) mol\u00adecules (A and B) and one half-mol\u00adecule of melamine (M). The other half of the melamine mol\u00adecule is generated by a twofold axis. 5FC mol\u00adecules A and B are linked through two different homosynthons [R22(8) ring motif]; one is formed via a pair of N\u2014H\u22efO hydrogen bonds and the second via a pair of N\u2014H\u22efN hydrogen bonds. In addition to this pairing, the O atoms of 5FC mol\u00adecules A and B inter\u00adact with the N2 amino group on both sides of the melamine mol\u00adecule, forming a DDAA array of quadruple hydrogen bonds and generating a supra\u00admolecular pattern. The 5FC (mol\u00adecules A and B) and two melamine mol\u00adecules inter\u00adact via N\u2014H\u22efO, N\u2014H\u22efN and N\u2014H\u22efO, N\u2014H\u22efN, C\u2014H\u22efF hydrogen bonds forming R66(24) and R44(15) ring motifs. The crystal structure is further strengthened by C\u2014H\u22efF, C\u2014F\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions.The asymmetric unit of the title compound, 4C The twofold axis of melamine coincides with the crystallographic twofold axis. An ORTEP view of the crystal structure is shown in Fig.\u00a01A and 1.3492\u2005(18)\u2005\u00c5 in 5FC B and the corresponding internal angles at the carbon-carrying fluorine atom [C2A\u2014N3A\u2014C4A = 119.96\u2005(13) in 5FC A and C2B\u2014N3B\u2014C4B = 119.92\u2005(13)\u00b0 in 5FC B] agree with those reported in the literature mol\u00adecules are involved in the generation of a quadruple hydrogen-bonded DDAA array having a fused-ring sequence of via N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds. These quadruple arrays are further linked by three large ring motifs: A mol\u00adecules, two 5FC B mol\u00adecules and two melamine mol\u00adecules through several N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, generating a hexa\u00admeric supermolecule. The A mol\u00adecule, two 5FC B mol\u00adecules and one melamine mol\u00adecule through N\u2014H\u22efO, N\u2014H\u22efN and C\u2014H\u22efF hydrogen bonds, generating a tetra\u00admeric supermolecule. Similarly, the A mol\u00adecules, one 5FC B mol\u00adecule and one melamine mol\u00adecule through N\u2014H\u22efO, N\u2014H\u22efN and C\u2014H\u22efF hydrogen bonds, generating another tetra\u00admeric supermolecule. The association of these DDAA array and A and B mol\u00adecules with an inter\u00adplanar distance of 3.475\u2005(6)\u2005\u00c5, centroid-to-centroid distance of 3.6875\u2005(11)\u2005\u00c5, and slip angle of 19.52\u00b0. The crystal structure is further strengthened by a C\u2014F\u22ef\u03c0 inter\u00adaction [3.4541\u2005(14)\u2005\u00c5] between 5-fluoro\u00adcytosinium mol\u00adecule A and the melamine mol\u00adecule while the melamine mol\u00adecule inter\u00adacts with them via N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, generating the supra\u00admolecular architecture.In this co-crystal, 5FC mol\u00adecules et al., 1982et al., 2009et al., 2013abet al., 2012et al., 2013et al., 2001et al., 2014abet al., 2005et al., 2007The crystal structure of 5-fluoro\u00adcytosine monohydrate (Louis Hot aqueous solutions of 5-fluoro\u00adcytosine (32\u2005mg) and melamine (31\u2005mg) were mixed in a 1:1 molar ratio. The resulting solution was warmed to 353\u2005K using a water bath for half an hour and kept at room temperature for crystallization. After one week, colourless crystals were obtained.A, N4B) groups were located in a difference Fourier map and refined freely. The other hydrogen atoms were positioned geometrically and were refined using a riding model with Uiso(H) = 1.2Ueq(parent atom).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901600476X/hg5470sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S205698901600476X/hg5470Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901600476X/hg5470Isup3.cmlSupporting information file. DOI: 1469709CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit is completed by one-half of the 2-({4-[(carboxyl\u00adatometh\u00adyl)carbamo\u00adyl]phen\u00adyl}formamido)\u00adacetate dianion, which is located on a centre of inversion, and by three water mol\u00adecules. Two [Co(CO3)(C10H8N2)2]+ cations are connected through C\u2014H\u22efO contacts by the uncoordinating anions. The aromatic rings of the 2,2\u2032-bi\u00adpyridine ligands and di\u00adacetate anions are involved in \u03c0\u2013\u03c0 stacking and C\u2014H\u22ef\u03c0 inter\u00adactions. The centroid\u2013centroid distances are in the range 3.4898\u2005(4)\u20133.6384\u2005(5)\u2005\u00c5. The crystal structure is stabilized by further O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, which give rise to a three-dimensional supra\u00admolecular network.The complex cation of the title compound, [Co(CO DOI: 10.1107/S160053681400631X/wm5010Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S160053681400631X/wm5010Isup3.cdxSupporting information file. DOI: 992984CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Its geometry is different from saturated peptides but is in excellent agreement with other de\u00adhydro\u00adalanine compounds. In the crystal, an N\u2014H\u22efO hydrogen bond links the mol\u00adecules in a herringbone packing arrangement.In the crystal structure of the de\u00adhydro\u00addipeptide (Boc-Phe-\u0394Ala-OiPr), the mol\u00adecule has a 20H28N2O5), the mol\u00adecule has a trans conformation of the N-methyl\u00adamide group. The geometry of the de\u00adhydro\u00adalanine moiety is to some extent different from those usually found in simple peptides, indicating conjugation between the H2C=C group and the peptide bond. The bond angles around de\u00adhydro\u00adalanine have unusually high values due to the steric hindrance, the same inter\u00adaction influencing the slight distortion from planarity of the de\u00adhydro\u00adalanine. The mol\u00adecule is stabilized by intra\u00admolecular inter\u00adactions between the isopropyl group and the N atoms of the peptide main chain. In the crystal, an N\u2014H\u22efO hydrogen bond links the mol\u00adecules into ribbons, giving a herringbone head-to-head packing arrangement extending along the [100] direction. In the stacks, the mol\u00adecules are linked by weak C\u2014H\u22efO hydrogen-bonding associations.In the title compound, the de\u00adhydro\u00addipeptide (Boc\u2013Phe\u2013\u0394Ala\u2013OiPr, C The unsaturated amino acid is introduced into the structure of these polycyclic peptides by post-translational modification of selected serine and threonine residues -phenyl\u00adalanylde\u00adhydro\u00adalanine isopropyl ester and its structure determination by single-crystal X-ray crystallographic methods are presented.De\u00adhydro\u00adpeptides are a class of compounds containing at least one residue of an \u03b1,\u03b2-de\u00adhydro\u00adamino acid. These compounds are of inter\u00adest in many fields of science because of their structural and chemical properties. De\u00adhydro\u00adamino acids are found in natural products (Bonauer N-(tert-but\u00adoxy\u00adcarbon\u00adyl)phenyl\u00adalanylde\u00adhydro\u00adalanine isopropyl ester is shown in Fig.\u00a01trans-conformation of the N-methyl\u00adamide group. The geometry of the de\u00adhydro\u00adalanine is to some extent different from those usually found in simple peptides glycine , C17\u2014N19\u2014C20 = 126.8\u2005(2) and O18\u2014C17\u2014N19 = 123.5\u2005(2)\u00b0] due to the steric hindrance between atoms C21 and O18. The same inter\u00adaction influences the slight distortion from planarity of the de\u00adhydro\u00adalanine moiety. The \u03c9, \u03d5 and \u03c8 torsion angles of the de\u00adhydro\u00adalanine residue are \u2212166.9\u2005(2), 175.1\u2005(2) and 178.0\u2005(2)\u00b0. The geom\u00adetries of the phenyl\u00adaniline and the protecting groups are normal. There are four intra\u00admolecular C\u2014H\u22efO close contacts but three of them have a D\u2014H\u22efA angle of less than 120\u00b0.The mol\u00adecular structure of i hydrogen bonds of Boc\u2013Phe\u2013\u0394Ala was dissolved in 5\u2005ml of methanol and calcium carbonate 0.329\u2005g (1\u2005mM) was added. The mixture was stirred for one\u2005h at room temperature, after which the solvent was evaporated. The residue was dissolved in 7\u2005ml of DMF and isopropyl iodide was added in portions to the stirred mixture at room temperature during the reaction, the progress of which was monitored by thin-layer chromatography, using 5% methanol in chloro\u00adform as eluent. After completion of the reaction, the solvent was evaporated and the oily residue was dissolved in ethyl acetate and washed consecutively with: 1 M HCl, saturated KHCO3, 0.1 M Na2S2O3 and brine. The organic layer was dried over anhydrous MgSO4 and the title compound was obtained in 81% yield (m.p. = 367\u2013369\u2005K). Recrystallization was performed using mixture of diethyl ether and hexane.The de\u00adhydro\u00addipeptide was obtained by condensation of al. 2008. For thi1H NMR \u03b4 1.26 , 1.30 , 2.76 , 3.02 , 4.27\u20134.39 , 5.01 , 5.70 , 6.23 , 7.15\u20137.36 , \u03b4 9.30 . 13C NMR \u03b4 21.43, 28.10, 36.63, 56.34, 69.40, 78.41, 108.65, 126.29, 128.07, 129.25, 132.71, 138.03, 155.53, 162.81, 171.53. IR 3600\u20132800 broad (H-bonding), 1715 (C=Oester), 1700 (C=Ourethane), 1690 IAB (C=Oamide), 1632 (C=C), 1526 IIAB (C\u2013N and N\u2013H), 1317 (CO\u2013N\u2013C=and N\u2013(C=C)\u2013CO), 1196 and 1166 (C\u2013O\u2013C), 896 (=CH2).Uiso (H) = 1.2Ueq(N), C\u2014Haromatic = 0.95\u2005\u00c5 and Uiso (H) = 1.2Ueq(C), C\u2014Hmeth\u00adyl = 0.98\u2005\u00c5 and Uiso (H) = 1.5Ueq(C); C\u2014Hmethyl\u00adene = 0.99\u2005\u00c5 or C\u2014Hmethine = 0.95\u2005\u00c5 and Uiso (H) = 1.2Ueq(C). Although not definitive, the absolute structure factor configuration, was \u22120.1\u2005(6) for 1095 Friedel pairs.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814025197/zs2321sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S1600536814025197/zs2321Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814025197/zs2321Isup3.cmlSupporting information file. DOI: 1034604CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Packing in the title keto ester compound is dominated by the formation of inversion dimers by both non-classical hydrogen bonds and offset \u03c0\u2013\u03c0 stacking inter\u00adactions. 15H10BrClO3, was synthesized in a single-step reaction by condensation of 3-bromo\u00adbenzoic acid with 2-bromo-1-(4-chloro\u00adphen\u00adyl)ethanone in di\u00admethyl\u00adformamide in the presence of tri\u00adethyl\u00adamine as a catalyst. The structure consists of an aryl ketone moiety linked to an aryl ester unit by a methyl\u00adene group. Both units are reasonably planar and are almost orthogonal, with an angle of 88.60\u2005(3)\u00b0 between them. In the crystal, mol\u00adecules form five separate sets of inversion dimers. Three of these are generated by two C\u2014H\u22efO inter\u00adactions and a C\u2014H\u22efBr contact, and form chains along c and along the ab cell diagonal. In addition, two inversion-related \u03c0\u2013\u03c0 stacking inter\u00adactions between like aryl rings again form chains of mol\u00adecules but in this instance along the bc diagonal. These contacts generate infinite layers of mol\u00adecules parallel to (011) and stack the mol\u00adecules along the a-axis direction.2-(4-Chloro\u00adphen\u00adyl)-2-oxoethyl 3-bromo\u00adbenzoate, C There is an r.m.s. deviation of 0.119\u2005\u00c5 from the best-fit plane through atoms Br1, C1\u2013C8, O1, O2 [maximum deviation 0.2477\u2005(11)\u2005\u00c5 for O1] while the plane of the carboxyl\u00adate unit subtends an angle of 15.5\u2005(2)\u00b0 to that of the bromo\u00adbenzene ring. In addition, the plane of the aryl ketone unit C8\u2013C15, O3, Cl1 has an r.m.s. deviation of 0.010\u2005\u00c5 [maximum deviation 0.0171\u2005(15)\u2005\u00c5 for C15]. The aryl ketone and aryl ester planes are almost orthogonal with an angle of 88.61\u2005(3)\u00b0 between them. Bond lengths and angles in the mol\u00adecule are normal and are generally similar to those found in closely related mol\u00adecules \u2005\u00c5, and neighbouring 4-chloro\u00adphenyl rings Cg2\u22efCg2v = 3.8585\u2005(11)\u2005\u00c5, in this case along the bc diagonal, Fig.\u00a03Cg1 and Cg2 are the centroids of the C1\u2013C6 and C10\u2013C15 rings, respectively; symmetry codes (iv) \u2212x, 2\u00a0\u2212\u00a0y, \u2212z; (v) 2\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z]. These contacts combine to generate extended layers of mol\u00adecules parallel to (011), Fig.\u00a04a-axis direction, Fig.\u00a05In the crystal structure, each mol\u00adecule forms five separate inversion dimers. C8\u2014H8et al., 2011aet al., 2011b Chidan Kumar et al., 2014abet al., 2014cA search of the Cambridge Crystallographic Database , followed by a solution of 2-bromo-1-(4-chloro\u00adphen\u00adyl)ethanone (1.0\u2005mmol). The reaction mixture was stirred for 2\u2005h. Progress of the reaction was monitored by TLC. After completion, the mixture was poured into water and the precipitated solid was filtered, dried and recrystallized (EtOAc/hexa\u00adne) to afford 2-(4-chloro\u00adphen\u00adyl)-2-oxoethyl 3-bromo\u00adbenzoate (1). The formation of keto ester (3) was indicated by the appearance of two typical stretching vibrations \u03bd(C=O) ester (1724) and \u03bd(C=O) keto (1698)\u2005cm\u22121, respectively and the disappearance of characteristic IR stretching absorptions ascribable to the carb\u00adoxy\u00adlic acid group in the region of 3400\u20132400\u2005cm\u22121. In the 1H NMR spectrum, the signals for the aromatic protons appeared in their respective regions and the disappearance of a characteristic signal for the COOH proton confirmed the formation of the title compound (1). The 13C NMR spectrum displayed two characteristic signals for the keto and ester carbonyl carbon atoms at 190.7 and 165.3 p.p.m., respectively. Yield: 88%; m.p. 372\u2013373\u2005K; Rf: 0.72 (10% EtOAc/hexa\u00adne); IR : 3089 (Csp2-H), 2933, 2856 (Csp3-H), 1724 (C=O ester), 1698 (C=O ketone), 1585, 1479 (C=C Ar), 1225 (C\u2014O); 1H NMR : \u03b4 8.06\u20138.03 , 7.94\u20137.90 , 7.73\u20137.70 , 7.52\u20137.48 , 7.46\u20137.36 , 5.57 ; 13C NMR : \u03b4 190.7, 165.3, 140.6, 134.5, 133.1, 132.4, 132.0, 131.0, 129.3, 129.3, 127.3, 122.1, 66.5.The preparation followed a procedure developed for the preparation of a related compound (Khan Uiso(H) = 1.2Ueq(C) for aromatic, and C\u2014H = 0.99\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for the methyl\u00adene H atoms.All H atoms were refined using a riding model, with C\u2014H = 0.95\u2005\u00c5 and 10.1107/S1600536814021643/hg5410sup1.cifCrystal structure: contains datablock(s) global, 1. DOI: 10.1107/S1600536814021643/hg54101sup2.hklStructure factors: contains datablock(s) 1. DOI: 864789CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compounds, N\u2014H\u22efO hydrogen bonds lead to dimers; the dimers are linked by weak inter\u00adactions into a three-dimensional network in one case and chains in the other. N-(aryl\u00adsulfon\u00adyl)aryl\u00adamides, namely N-(3-fluoro\u00adbenzo\u00adyl)benzene\u00adsulfonamide, C13H10FNO3S, (I), and N-(3-fluoro\u00adbenzo\u00adyl)-4-methyl\u00adbenzene\u00adsulfonamide, C14H12FNO3S, (II), are described and compared with related structures. The dihedral angle between the benzene rings is 82.73\u2005(10)\u00b0 in (I) compared to 72.60\u2005(12)\u00b0 in (II). In the crystal of (I), the mol\u00adecules are linked by C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions, resulting in a three-dimensional grid-like architecture, while C\u2014H\u22efO inter\u00adactions lead to one-dimensional ribbons in (II). The crystals of both (I) and (II) feature strong but non-structure-directing N\u2014H\u22efO hydrogen bonds with R22(8) ring motifs. The structure of (I) also features \u03c0\u2013\u03c0 stacking inter\u00adactions.The crystal structures of two N-(Aryl\u00adsulfon\u00adyl)aryl\u00adamides have received much attention as they constitute an important class of drugs for Alzheimers disease aryl\u00adamides are known to be potent anti\u00adtumour agents against a broad spectrum of human tumour xenografts in nude mice syn to the N\u2014H bond in the central \u2013C\u2014SO2\u2014N\u2014C(=O)\u2013 segment. By contrast, in (II)anti with respect to the meta-fluoro substitution on the benzoyl ring. The dihedral angle between the benzene rings is 82.73\u2005(10)\u00b0 in (I)The C7 chains, forming columns propagating along the b-axis direction inter\u00adactions link the mol\u00adecules into chains along the c axis. These chains are inter\u00adconnected via C2\u2014H2\u22ef\u03c0ar\u00adyl (\u03c0 system of the sulfonyl\u00adbenzene ring) and C11\u2014H11\u22ef\u03c0ar\u00adyl (\u03c0 system of the sulfonyl\u00adbenzene ring) inter\u00adactions, forming a three-dimensional grid-like structure rs Fig.\u00a03. These don Fig.\u00a03. In addire Fig.\u00a04. The cryvia C13\u2014H13\u22efO1 inter\u00adactions, as observed in (I)via C5\u2014H5\u22efO3 inter\u00adactions into ac plane rs Fig.\u00a05. The molne Fig.\u00a05. The oveN-(aryl\u00adsulfon\u00adyl)aryl\u00adamides, namely N-(benzo\u00adyl)benzene\u00adsulfonamide (III), N-(3-chloro\u00adbenzo\u00adyl)benzene\u00adsulfonamide (IV), N-(3-methyl\u00adbenzo\u00adyl)benzene\u00adsulfonamide (V), N-(benzo\u00adyl)-4-methyl\u00adbenzene\u00adsulfon\u00adamide (VI) and N-(3-methyl\u00adbenzo\u00adyl)-4-meth\u00adylbenzene\u00adsulfonamide (VII) have previously been reported. A comparison of the dihedral angle between the two benzene rings in these closely related structures indicates that introducing a methyl substituent into the para position of the benzene\u00adsulfonyl ring lowers the dihedral angle with compound (VII) being an exception. The dihedral angle values are 80.3\u2005(1)\u00b0 in (III) C(4) chains via strong structure-directing N\u2014H\u22efO hydrogen bonds. The structures do not feature any other type of inter\u00adactions. However, in (I)ar\u00adyl. Furthermore, introducing the methyl substit\u00aduent into the benzene\u00adsulfonyl ring of (I)The crystal structures of five related Compounds (I)Uiso = 1.2 or 1.5Ueq(parent atom). To improve considerably the values of R1, wR2 and GOOF, reflections with very bad agreement (\u221220 0 0), (\u221220 0 10) and (\u221219 1 15) in (I)Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016003248/hb7565sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989016003248/hb7565Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016003248/hb7565IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989016003248/hb7565Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016003248/hb7565IIsup5.cmlSupporting information file. DOI: 1418689, 1418688CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A and B) in the asymmetric unit, which differ essentially in the orientation of the terminal amino\u00adphenyl ring with respect to the central benzene ring. In the crystal, mol\u00adecules are linked via N\u2014H\u22efO hydrogen bonds forming \u2013A-B\u2013A\u2013B\u2013 zigzag chains propagating along [010].The title compound crystallized with two independent mol\u00adecules in the asymmetric unit. There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond in each mol\u00adecule with the phenol ring being inclined to the central benzene ring by 4.93\u2005(14) and 7.12\u2005(14)\u00b0 in mol\u00adecules A and B, respectively. The conformation of the two mol\u00adecules differs essentially in the orientation of the terminal amino\u00adphenyl ring with respect to the central benzene ring; this dihedral angle is 50.51\u2005(4)\u00b0 in mol\u00adecule A and 54.61\u2005(14)\u00b0 in mol\u00adecule B. The two outer aromatic rings are inclined to one another by 51.39\u2005(14) and 49.88\u2005(14)\u00b0 in mol\u00adecules A and B, respectively. In the crystal, mol\u00adecules are connected by N\u2014H\u22efO hydrogen bonds generating \u2013A-B\u2013A\u2013B\u2013 zigzag chains extending along [010]. The chains are linked via C\u2014H\u22ef\u03c0 inter\u00adactions involving neighbouring A mol\u00adecules, forming slabs lying parallel to (100).The title compound, C In mol\u00adecules A and B the phenol rings (C1\u2013C6 and C20\u2013C25) are inclined to the central benzene rings (C8\u2013C13 and C27\u2013C32) by 4.93\u2005(14) and 7.12\u2005(14)\u00b0, respectively.The title compound crystallized with two independent mol\u00adecules \u00b0 in mol\u00adecule B. The two outer aromatic rings are inclined to one another by 51.39\u2005(14) and 49.88\u2005(14)\u00b0 in mol\u00adecules A and B, respectively. The C\u2014N, C\u00a0N and C\u2014C bond lengths are normal and close to the values observed in related structures with respect to the central benzene rings (C8\u2013C13 and C27\u2013C32); this dihedral angle is 50.51\u2005(4)\u00b0 in mol\u00adecule A-B\u2013A\u2013B\u2013 zigzag chains extending along [010]; Table\u00a01via C\u2014H\u22ef\u03c0 inter\u00adactions involving neighbouring A mol\u00adecules, forming slabs lying parallel to (100); see Table\u00a01In the crystal, mol\u00adecules are connected by N\u2014H\u22efO hydrogen bonds, generating \u2013et al., 2013et al., 2012viz. N-[(E)-4-chloro\u00adbenzyl\u00adidene]-N\u2032-phenyl\u00adbenzene-1,4-di\u00adamine (II) \u00b0 in mol\u00adecule B, owing to the presence of the intra\u00admolecular O\u2014H\u22efN hydrogen bond. The outer phenyl ring is inclined to the central six-membered ring by 44.18\u2005(11)\u00b0 in (II), compared to 50.51\u2005(4) and 54.61\u2005(14)\u00b0 in mol\u00adecules A and B, respectively, of the title compound.There are very few examples of similar compounds in the literature although some metal complexes of similar ligands have been reported on of salicyladehyde in 5\u2005ml of absolute ethanol was added dropwise with stirring. The mixture was stirred for 10\u2005min, two drops of glacial acetic acid were then added and the mixture was further refluxed for 2\u2005h. The resulting reddish yellow precipitate was recovered by filtration, washed several times with a small portions of EtOH and then with diethyl ether to give 120\u2005mg (75%) of the title compound. Crystals suitable for X-ray analysis was obtained within 3 days by slow evaporation of a solution in methanol.100\u2005mg (1\u2005mmol) of Uiso(H) = 1.2Ueq(N) and = 1.5Ueq(O). All C-bound H atoms were positioned geometrically and refined using a riding model with C\u2014H = 0.93\u2005\u00c5 and with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989014026309/su5028sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989014026309/su5028Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989014026309/su5028Isup3.cmlSupporting information file. DOI: 1036844CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atom is located on a center of symmetry with an overall octa\u00adhedral coordination environment. Both intra- and inter\u00admolecular inter\u00adactions occur between the amino and acetate groups, leading to a layered structure.In a cadmium complex incorporating 1,3-di\u00adamino\u00adpropane and nitro\u00adphenyl\u00adacetate ligands, the Cd 8H6NO4)2(C3H10N2)2], the CdII atom is located on a center of symmetry with one independent Cd\u2014O distance of 2.3547\u2005(17)\u2005\u00c5 and two Cd\u2014N distances of 2.3265\u2005(18) and 2.3449\u2005(19)\u2005\u00c5. The CdII atom has an overall octa\u00adhedral coordination environment. Several types of hydrogen-bonding inter\u00adactions are evident. Both intra- and inter\u00admolecular inter\u00adactions occur between the amino groups and the O atoms of the acetate group. These N\u2014H\u22efO hydrogen bonds lead to a layered structure extending parallel to the bc plane. In addition, weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions involving the nitro groups exist, leading to the formation of a three-dimensional network structure.In the structure of the title compound, [Cd(C Several other weak inter\u00admolecular hydrogen-bonding inter\u00adactions of the C\u2014H\u22efO type also exist in the structure involving the O atoms of nitro groups and neighboring C\u2014H groups.Somewhat weaker inter\u00admolecular N\u2014H\u22efO inter\u00adactions involving the same types of donor and acceptor groups occur between neighboring mol\u00adecules Table\u00a01 and leadne Fig.\u00a02. It shou2, 0.4\u2005mmol (29.7\u2005mg) of 1,3-di\u00adamino\u00adpropane, and 0.4\u2005mmol (72.5\u2005mg) of 4-nitro\u00adphenyl\u00adacetic acid were added to 2\u2005ml of methanol in a 5\u2005ml beaker. The sample was covered with aluminum foil containing several small vent holes and left for a week to evaporate. The slow evaporation method was used to crystallize a colorless mononuclear species and crystals were gathered for X-ray crystallographic analysis.0.2\u2005mmol (36.7\u2005mg) of anhydrous CdClUiso(H) = 1.2Ueq(C) and C\u2014H distances of 0.93\u2005\u00c5 for aromatic hydrogen atoms, Uiso(H) = 1.2Ueq(C) and C\u2014H distances of 0.97\u2005\u00c5 for methylene hydrogen atoms, and Uiso(H) = 1.2Ueq(N) and N\u2014H distances of 0.90\u2005\u00c5 for amino hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016000943/wm5258sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016000943/wm5258Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016000943/wm5258Isup3.cdxSupporting information file. DOI: 1447705CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, pairs of inversion-related mol\u00adecules are linked into inversion dimers 23H19NO2, an oxazine Mannich base derivative, the oxazine ring has a half-chair conformation. The 2-hy\u00addroxy\u00adnaphthalen-1-yl substituent is placed in an axial position. There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond, forming an S(6) graph-set motif. In the crystal, mol\u00adecules are connected by a pair of C\u2014H\u22ef\u03c0 inter\u00adactions into an inversion dimer, which is reinforced by another pair of weak C\u2014H\u22ef\u03c0 inter\u00adactions. The dimers are linked by a \u03c0\u2013\u03c0 inter\u00adaction [centroid-centroid distance = 3.6268\u2005(17)\u2005\u00c5], consolidating a column along the a axis. Furthermore, the columns inter\u00adact with each other by a weak C\u2014H\u22ef\u03c0 inter\u00adaction, generating a three-dimensional network.In the title compound, C Recently, we synthesized the title compound by a reaction between the cyclic aminal 1,3,6,8-tetra\u00adaza\u00adtri\u00adcyclo\u00adundecane (TATU) with 2-naphthol solvent-free at low temperature. Because a wide range of cured properties can be obtained \u2005\u00c5, \u03b8 = 50.0\u2005(3)\u00b0 and \u03c6 = 98.3\u2005(4)\u00b0 for the ring O1/C1/N1/C2/C12/C11. The methyl group bonded to atom N1 of the oxazine ring is placed in an axial position. The pendant naphthyl group (C21\u2013C30) makes a dihedral angle of 59.94\u2005(4)\u00b0 with the oxazine ring plane defined by atoms C11, C12 and O1. The bond lengths, N1\u2014C1 and O1\u2014C1, are normal and comparable to the corresponding values observed in the related structure of 6-bromo-2,4-bis\u00ad(3-meth\u00adoxy-phen\u00adyl)-3,4-di\u00adhydro-2H-1,3-naphthoxazine graph-set motif, where the N\u22efO distance is longer by about 0.04 and 0.03\u2005\u00c5, respectively, than the observed values in related structures of 1-(piperidin-1-ylmeth\u00adyl)-2-naphthol \u2005\u00c5 . Neighboring columns are connected by a weak C\u2014H\u22ef\u03c0 inter\u00adaction were manually ground together, heated to 313\u2005K and stirred for 12\u2005h under solvent-free conditions. Progress of the reaction was determined by TLC monitoring. After completion of the reaction, the mixture was cooled to room temperature and the solid residue was purified by silica gel column chromatography with benzene\u2013ethyl acetate (4:1) as the eluent to give 1-{oxazin-2(3H)-yl]meth\u00adyl}naphthalen-2-ol as a brown solid in 28% yield. This compound was obtained in its crystalline form by recrystallization from an absolute ethanol solution (m.p. 443\u2005K).2-Naphthol and 1,3,6,8-tetra\u00adaza\u00adtri\u00adcyclo\u00ad[4.3.1.1Uiso(H) value and the C\u2014C\u2014O\u2014H torsion angle were refined. C-bound H atoms were fixed geometrically (C\u2014H = 0.95 or 0.99\u2005\u00c5) and treated as riding with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015015583/is5412sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015015583/is5412Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015015583/is5412Isup3.cmlSupporting information file. DOI: 1419687CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In this complex, all CuII atoms have a square-pyramidal coordination sphere, with long axial and short basal Cu\u2014O distances.The dinuclear title compound crystallizes as a dimer forming a tetra\u00adnuclear copper(II) complex, [Cu 4(CH3O)4(C11H13O2)4], consists of dimeric dinuclear copper(II) complexes oriented around a centre of inversion. Within each dinuclear fragment, the two CuII atoms are in a distorted square-planar coordination sphere. Two neighbouring fragments are linked by four apical Cu\u2014O contacts, yielding an overall square-pyramidal coordination environment for each of the four CuII atoms. The mol\u00adecules are arranged in layers parallel to (101). Non-classical C\u2014H\u22efO hydrogen-bonding inter\u00adactions are observed between the layers.The structure of the title compound, [Cu The distances and coordination modes between CuII atoms vary and thus, the compound is a suitable study case for investigating different spin-coupling paths. This knowledge is deemed important for the design of tailor-made magnetic compounds.The title compound was obtained as a by-product in the synthesis of an unsymmetrically substituted copper(II) salophene complex ions Cu1 and Cu2 within the binuclear fragment is 2.9938\u2005(2)\u2005\u00c5 title compound consists of two dinuclear complex fragments oriented around a centre of inversion. Within each fragment the two Cu\u2005\u00c5 Fig.\u00a01 which isi bond to the methoxido ligand of the neighbouring fragment. For the Cu2 atom, the situation is comparable, with slightly shorter Cu\u2014O distances in comparison with Cu1: Cu2\u2014O3 1.8939\u2005(8)\u2005\u00c5, Cu2\u2014O4 1.9473\u2005(9)\u2005\u00c5, Cu2\u2014O5 1.9455\u2005(8)\u2005\u00c5 and Cu2\u2014O6 1.9081\u2005(8)\u2005\u00c5. The longer distance Cu2\u2014O1i of 2.4994\u2005(9)\u2005\u00c5 to the phenoxido ligand atom of the neighbouring fragment causes less sterical congestions at the Cu2 atom and thus, appears to be the cause for the shorter basal Cu\u2014O distances.Short distances Cu1\u2014O1 of 1.9166\u2005(8)\u2005\u00c5, Cu1\u2014O2 of 1.9557\u2005(9)\u2005\u00c5, Cu1\u2014O5 of 1.9522\u2005(8)\u2005\u00c5 and Cu1\u2014O6 of 1.9154\u2005(9)\u2005\u00c5 are found for the Cu1 atom to the basal O atoms within the binuclear fragment. A substanti\u00adally longer distance of 2.3703\u2005(9)\u2005\u00c5 is observed for the apical Cu1\u2014O52versus \u03bc3) of the two methoxido ligands in each fragment can be distinguished by the angles C24\u2014O5\u2014O6 versus C23\u2014O6\u2014O5 , resulting in a more pyramidal-like geometry. This differs to the more trigonal-planar geometry of O6 (see Table\u00a01versus 1.2963\u2005(13)\u2005\u00c5].The binding modes \u00b0 due to repulsion of the In the crystal, the tetra\u00adnuclear mol\u00adecules arrange in layers parallel to (101) Fig.\u00a02. Weak notert-butyl\u00adsalicyl\u00adaldehyde in 10\u2005ml THF, the mixture was stirred for 22\u2005h at room temperature. Addition of hexane yielded the title compound as a dark crystalline material from the reaction mixture .After treatment of 102\u2005mg (0.35\u2005mmol) 4-Br-salicyl-2-(2-amino)\u00adphenyl\u00adimine with 113\u2005mg of copper(II)acetate monohydrate (0.445\u2005mmol), 1\u2005ml tri\u00adethyl\u00adamine in 10\u2005ml THF, and 65.5\u2005mg (0.368\u2005mmol) 4-Uiso(H) = 1.5Ueq(C) for sp2 C atoms and of 0.98\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for sp3 C atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901500376X/wm5122sup1.cifCrystal structure: contains datablock(s) general, I. DOI: 10.1107/S205698901500376X/wm5122Isup2.hklStructure factors: contains datablock(s) I. DOI: 1050914CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Single crystals of the ligand have been grown and analyzed.Hydrazones and their metal complexes were heavily studied due to their pharmacological applications such as antimicrobial, anticonvulsant analgesic, anti-inflammatory and anti-cancer agents. This work aims to synthesize and characterize novel complexes of VO2+, Co2+, Ni2+, Cu2+, Zn2+, Zr4+ and Pd2+ complexes have the formulas: [VO(OBH\u2013H)2]\u00b7H2O, [Co(OBH)2Cl]Cl\u00b7\u00bdEtOH, [Ni2(OBH)Cl4]\u00b7H2O\u00b7EtOH, [Cu(OBH)2Cl2]\u00b72H2O, [Zn(OBH\u2013H)2], [Zr(OBH)Cl4]\u00b72H2O, and [Pd2(OBH)(H2O)2Cl4]\u00b72H2O. All complexes are nonelectrolytes except [Co(OBH)2Cl]Cl\u00b7\u00bdEtOH. OBH ligates as: neutral tetradentate (NNOO) in the Ni2+ and Pd2+ complexes; neutral bidentate (OO) in [Co(OBH)2Cl]Cl\u00b7\u00bdEtOH, [Zr(OBH)Cl4]\u00b72H2O and [Cu(OBH)2Cl2]\u00b72H2O and monobasic bidentate (OO) in the Zn2+ and VO2+ complexes. The NMR (1H and 13C) spectra support these data. The results proved a tetrahedral for the Zn2+ complex; square-planar for Pd2+; mixed stereochemistry for Ni2+; square-pyramid for Co2+ and VO2+ and octahedral for Cu2+ and Zr4+ complexes. The TGA revealed the outer and inner solvents as well as the residual part. The molecular modeling of [Ni2(OBH)Cl4]\u00b7H2O\u00b7EtOH and [Co(OBH)2Cl]Cl\u00b7\u00bdEtOH are drawn and their molecular parameters proved that the presence of two metals stabilized the complex more than the mono metal. The complexes have variable activities against some bacteria and fungi. [Zr(OBH)Cl4]\u00b72H2O has the highest activity. [Co(OBH)2Cl]Cl\u00b7\u00bdEtOH has more activity against Fusarium.Oxalo bis [OBH] has a monoclinic crystal with P 1 21/n 1 space group. The VO2+ and Co2+ complexes have square-pyramid structure. [Cu(OBH)2Cl2]\u00b72H2O and [Ni2(OBH)Cl4]\u00b7H2O\u00b7EtOH decomposed to their oxides while [VO(OBH\u2013H)2]\u00b7H2O to vanadium. The energies obtained from molecular modeling calculation for [Ni2(OBH)Cl4]\u00b7H2O\u00b7EtOH are less than those for [Co(OBH)2Cl]Cl\u00b7\u00bdEtOH indicating the two metals stabilized the complex more than mono metal. The Co(II) complex is polar molecule while the Ni(II) is non-polar.Oxalo bis structure was proved by X-ray crystallography. It coordinates with some transition metal ions as neutral bidentate; mononegative bidentate and neutral tetradentate. The complexes have tetrahedral, square-planar and/or octahedral structures. The VOThe online version of this article (doi:10.1186/s13065-015-0135-y) contains supplementary material, which is available to authorized users. In the preparation of VO\u22123\u00a0mol\u00a0L\u22121 DMSO solution of the compounds was measured on Orion 3 STAB Conductivity Bridge. The IR spectra were recorded as KBr discs on a FT/IR-6300 type A (400\u20134000\u00a0cm\u22121). The electronic spectra of the complexes were recorded on a Cary 5 UV\u2013vis spectrophotometer, varian (200\u2013900\u00a0nm). The 1H NMR spectra of the ligand and the diamagnetic complexes were recorded in DMSO-d6, on a Bruker WP 200 SY Spectrometer (400\u00a0MHz) at room temperature using tetramethylsilane (TMS) as an external standard. The magnetic measurements were carried out on a Johnson-Matthey magnetic balance, UK. The TGA thermograms were recorded (25\u2013800\u00a0\u00b0C) on a Shimadzu TGA-60; the nitrogen flow and heating rate were 50\u00a0ml/min and 10\u00a0\u00b0C min\u22121, respectively. The X-ray single crystal diffraction data were collected on a Rigaku R-Axis Rapid diffractometer using filtered Mo-K \u03b1-radiation. The structure was solved by the direct methods and expanded using Fourier techniques at Kuwait University. The ligand and its complexes were investigated for antimicrobial activity against Bacillus, Aspergillus, Escherichia coli, Pennicillium and Fusarium as reported earlier \u00b7H2O, [Co(OBH)2Cl]Cl\u00b7\u00bdEtOH, [Cu(OBH)2Cl2]\u00b72H2O, [Zn(OBH\u2013H)2], [Zr(OBH)Cl4]\u00b72H2O and binuclear complexes: [Ni2(OBH)Cl4]\u00b7H2O\u00b7EtOH and [Pd2(OBH)(H2O)2Cl4]\u00b72H2O. All complexes are colored, solid and stable towards air and moisture at room temperature. They have high melting points and are insoluble in most common organic solvents and completely soluble in DMSO. The molar conductance values complex supports the formation of [Co(OBH)2Cl]+Cl\u2212\u00b7\u00bdEtOH and \u03bd(C=N) vibrations at 3325, 1701 and 1605, respectively in its IR spectrum Cl4]\u00b72H2O coordinating through the two amidic carbonyl groups based on the following observations: the \u03c5(C=O) band observed at 1701\u00a0cm\u22121 in ligand spectrum was shifted to 1686\u20131699\u00a0cm\u22121 in complexes having little intensity indicating that the two amidic carbonyl groups (C=Oamidic) participated in bonding while the other two carbonyl (C=Oketonic) still at the same position. The new band at 464\u2013495\u00a0cm\u22121 is due to \u03c5(M\u2013O) vibration showed splitting of NH signal as a result of conversion of one of NHC=O to N=C\u2013OH and the existence of the others without participation ketonic, (C=O)amidic free, (C=O)amidic bonded, (C=N), (C=N)*, (C\u2013O) and CH3, respectively. The appearance of (C=N)* (due to conversion of NHC=O to N*=C\u2013O) and (C\u2013O) peaks confirm enolization process and \u03c5(M\u2013ed water and abse2(OBH)Cl4]\u00b7H2O\u00b7EtOH (Structure\u00a02(OBH)(H2O)2Cl4]\u00b72H2O amidic to 1644 in the Pd(II) complex and to 1552 and 1676 in the Ni(II) complex together with appearance of \u03c5(M\u2013N) \u00b7H2O\u00b7EtOH has a value of 371.7 (the base peak) corresponding to Ni(OBH)Cl\u00b7\u00bdEtOH meaning that this species is highly stable. Multi peaks were observed ending with one at 128.9 (intensity 65\u00a0%) due to ZrO2.Moreover, the mass spectrum of \u00b7H2O\u00b7EtOH is 1.36 BM which is less than the normal values reported for tetrahedral or octahedral coordination containing two unpaired electrons. Its electronic spectrum showed a broad band at 19,050\u00a0cm\u22121 (\u025b\u00a0=\u00a0180\u00a0mol\u22121\u00a0L) typical of a square-planar structure with some distortion [2(OBH)(H2O)2Cl4]\u00b72H2O proved the square-pyramid structure in which the metal is surrounded by NO donors, two chloro and one coordinated water. The bands at 37,540 and 28,470\u00a0cm\u22121 are attributed to charge transfer transitions, probably O\u00a0\u2192\u00a0Pd transition \u00b7H2O exhibits one band with maximum at 20080\u00a0cm\u22121 assigned to the 2E2g\u00a0\u2192\u00a02T2g transition in an octahedral geometry [g level. The magnetic moment value (1.45 BM) was found lower than the values reported for the d9\u2013system containing one unpaired electron (1.73\u20132.25\u00a0BM) suggesting interactions between the copper centers.The electronic spectrum of [Cu(OBH)geometry . The ban2Cl]Cl\u00b7\u00bdEtOH showed three decomposition steps at mid- points of 60, 319 and 500\u00a0\u00b0C corresponding to the removal of \u00bdCl2\u00a0+\u00a0\u00bdEtOH ; C16H24N4O6Cl and C4H4N2 leaving [CoO4N2] moiety .The decomposition steps, the DTG maximum temperature and the removing species are shown in Table\u00a02]\u00b7H2O showed also three steps; the first (mid. point 56\u00a0\u00b0C) represents the removal of the outside water molecule ; the second (mid. point 289\u00a0\u00b0C) represents the loss of C16H24N4O6 and the third for the repulsion of C4H4N4O2 . The residue is vanadium metal .The TG curve of [VO(OBH-H)2Cl2]\u00b72H2O thermogram showed decomposition steps ending with copper oxide at Temp. >400\u00a0\u00b0C. The decomposition showed the removal of the two hydrated water in the first step at mid. point of 59\u00a0\u00b0C. The other two steps were observed at 291 and 374\u00a0\u00b0C corresponding to the removal of C16H24N4O6 and C4H4N4O3, respectively, leaving CuO as a residue.[Cu(OBH)2(OBH)Cl4]\u00b7H2O\u00b7EtOH showed four steps. The first at 72\u00a0\u00b0C is due to the removal of the outside water and EtOH . The second step (368\u00a0\u00b0C) represents the loss of Cl2\u00a0+\u00a0C8H12N2O2 . The third step represents the repulsion of Cl2 . The fourth step corresponding to the removal of C2H2N2. The residue is 2NiO . The first three steps observed at mid. points of 76, 313 and 449\u00a0\u00b0C are corresponding to the removal of (Cl2\u00a0+\u00a0H2O); (H2O\u00a0+\u00a0C8H12N2O2) and Cl2, respectively Cl2(OBH)(H2O)2Cl4]\u00b72H2O showed two main steps at 75 and 322\u00a0\u00b0C due to the liberation of the outside water and 2H2O\u00a0+\u00a02Cl2\u00a0+\u00a0C4H12, respectively. High residue \u00a0% was found over 500\u00a0\u00b0C.The thermogram of [Pd2Cl]Cl\u00b7\u00bdEtOH and [Ni2(OBH)Cl4]\u00b7H2O\u00b7EtOH (Structure\u00a02(OBH)Cl4]\u00b7H2O\u00b7EtOH are less than those of [Co(OBH)2Cl]Cl\u00b7\u00bdEtOH indicating that the presence of two metals stabilized the complex more than the mono metal lowering the energy. The dipole moment calculated for the Co(II) complex is 4.949\u00a0D proving the polar nature of the complex. The value of Ni(II) complex is 0.413\u00a0D indicating its non-polarity.Trials to grow single crystals for the investigated complexes were failed. In order to calculate the molecular parameters, \u00b72H2O has higher activity against all tested microorganisms except E. coli. The activity is highest and more with Penicillium (9\u00a0mm zone inhibition). The higher activity may be due the presence of non-ionizable chlorine and to the less planarity of the complex making it more lipophilic. Most compounds have high activity against Fusarium. [Cu(OBH)2Cl2]\u00b72H2O has higher value against Fusarium (15\u00a0mm). Comparing these data with that of ampicillin and those obtained for different hydrazone complexes showed more or less activity \u00b72H2O and [Ni2(OBH)Cl4]\u00b7H2O\u00b7EtOH decomposed to their oxides while [VO(OBH\u2013H)2]\u00b7H2O to the metal. The energies from molecular modeling calculation is less in [Ni2(OBH)Cl4]\u00b7H2O\u00b7EtOH than those for [Co(OBH)2Cl]Cl\u00b7\u00bdEtOH indicating that the presence of two metals stabilized the complex more than the mono metal. The Co(II) complex is polar molecule while the Ni(II) is non-polar.Oxalo bis has been prepared and characterized by x-ray crystallography. It coordinates as neutral bidentate; mononegative bidentate and neutral tetradentate. The complexes have tetrahedral, square-planar and/or octahedral structures. The VOCrystallographic data for the structure reported in this paper have been deposited with Cambridge Crystallographic Data Center as supplementary publication CCDC-985982."} +{"text": "This article describes data related to the research article titled \u201cFunctional characterization of importin \u03b18 as a classical nuclear localization signal receptor\u201d Specifications TableValue of the data\u2022These data are valuable to researchers interested in the molecular mechanisms by which the importin \u03b1-cNLS substrate complex dissociates in the nucleus.\u2022These data show that importin \u03b11 and \u03b18 have substantial differences in dimer-forming ability, despite both proteins belonging to the same subfamily.\u2022These data provide a new insight into the function of nuclear-localized importin \u03b1s.1To examine whether importin \u03b1s can directly bind with other importin \u03b1 subtypes, FLAG-tagged importin \u03b16, \u03b17, or \u03b18 recombinant proteins were incubated with either GST-importin \u03b18, or GST-importin \u03b11, and analyzed by western blotting A. To inv22.1EcoRI and NotI, and then subcloned into the pGEX6P3 plasmid . The construct integrity of pGEX6P3/FLAG-h-importin \u03b16 and pGEX6P3/FLAG-h-importin \u03b17 was confirmed by DNA sequencing using the following primers: pGEX 5\u2032 sequencing primer: 5\u2032-GGGCTGGCAAGCCACGTTTGGTG-3\u2032, pGEX 3\u2032 sequencing primer: 5\u2032-CCGGGAGCTGCATGTGTCAGAGG-3\u2032, importin \u03b16 sequencing primer: 5\u2032-GCATCTGGAACTTTTCTGCATACC-3\u2032, or importin \u03b17 sequencing primer: 5\u2032-GTACATTACAGTTTGAAGCTGCCT-3\u2032.The N-terminus FLAG-tagged cDNAs encoding full-length human importin \u03b16 or full-length human-importin \u03b17 were amplified from either HEK293 cells or MRK-nu-1 cells by PCR using the following primers: importin \u03b16 Forward: 5\u2032-CCCGAATTCCGCCATGGACTACAAGGACGACGACGACAAGATGGATGCCATGGCTAGTCC-3\u2032 and importin \u03b16 Reverse: 5\u2032-CCCGCGGCCGCCTCGAGTTAAAGTTGAAATCCATCC-3\u2032 or importin \u03b17 Forward: 5\u2032-CCCGAATTCCGCCATGGACTACAAGGACGACGACGACAAGATGGAGACCATGGCGAGC-3\u2032 and importin \u03b17 Reverse: 5\u2032-CCCGCGGCCGCCTCGAGTTATAGCTGGAAGCCCTCC-3\u2032. The PCR program was as follows: 2\u00a0min at 94\u00a0\u00b0C followed by 40 cycles of 30\u00a0s at 98\u00a0\u00b0C and 15\u00a0min at 68\u00a0\u00b0C. The PCR products were digested with EcoRI and NotI sites of pGEX6P3, and then sequenced using either the pGEX 5\u2032 or pGEX 3\u2032 sequencing primer and the importin \u03b18 sequencing primer: 5\u2032-CAACATCGCTTCAGGGACTTCG-3\u2032.The human-importin \u03b18 cDNA with the FLAG-tag at the N-terminus was amplified by PCR from the pcDNA5/3xFLAG-h-importin \u03b18 plasmid The construct encoding SV40 large T antigen NLS was subcloned from pGEX2T-SV40TNLS-GFP BamHI and XhoI sites of pGEX6P1, and then verified by sequencing.The human cDNA encoding the tumor protein p53 (NM_000546) was amplified from MCF7 cells by PCR performed using the following primers: p53 Forward: 5\u2032-CACGGATCCATGGAGGAGCCGCAGTCAGATC-3\u2032 and p53 Reverse: 5\u2032-GGACTCGAGTCAGTCTGAGTCAGGCCCTTCTG-3\u2032. The PCR program was as follows: one cycle of 2\u00a0min at 94\u00a0\u00b0C; 40 cycles of 15\u00a0s at 94\u00a0\u00b0C, 30\u00a0sec at 64\u00a0\u00b0C, and 1\u00a0min 20\u00a0s at 68\u00a0\u00b0C; and one cycle of 10\u00a0min at 68\u00a0\u00b0C. The PCR product was subcloned into the 2.2Escherichia coli Rosetta, and then the cells were grown at 37\u00a0\u00b0C in LB medium containing 50\u00a0\u03bcg/mL ampicillin. Expression was induced by addition of 1\u00a0mM isopropyl-\u03b2-d-thiogalactopyranoside (IPTG), followed by incubation at 20\u00a0\u00b0C for 12\u00a0h. The bacteria were lysed in lysis buffer , 0.2\u00a0mM phenylmethylsulfonyl fluoride (PMSF), 1\u00a0\u03bcg/mL aprotinin , 1\u00a0\u03bcg/mL pepstatin , and 1\u00a0\u03bcg/mL leupeptin (Peptide Institute)) by freeze\u2013thawing twice and passing through a French press . The cell lysates were sonicated using a Sonifier 250 , and centrifuged at 20,400g at 4\u00a0\u00b0C for 30\u00a0min. The resultant supernatant was incubated with glutathione-Sepharose 4B beads at 4\u00a0\u00b0C for 12\u00a0h. After the GSH beads were washed five times with lysis buffer, GST-tagged proteins were eluted with elution buffer . Cleavage of GST from the GST-fused protein was performed using PreScission protease with 10\u00a0units/mg of fusion protein at 4\u00a0\u00b0C for 12\u00a0h in cleavage buffer . Finally, the purified proteins were dialyzed against dialysis buffer and concentrated by ultrafiltration using Amicon Ultra centrifugal filter units .Recombinant proteins fused to GST were purified as follows: The expression vectors were transformed into 2.32.4The following antibodies were used for western blotting: anti-FLAG M2 antibody , anti-GST antibody , anti-GFP antibody , anti-p53 (FL-393) antibody , and horseradish peroxidase (HRP)-conjugated anti-mouse or anti-rabbit IgG secondary antibodies 2.5http://rsbweb.nih.gov/ij/).Samples were loaded on a 10% SDS-PAGE gel, and the separated proteins in the gel were transferred onto an Immobilon-P membrane using a semi-dry transfer blotting system . The membrane was blocked with blocking buffer consisting of 3% skim milk in Tris-buffered saline with 0.05% Tween (TBS-T) for 1\u00a0h. The membrane was probed with primary antibodies diluted in Can Get Signal Immunoreaction Enhancer Solution 1 at 4\u00a0\u00b0C overnight, and then incubated with the HRP-conjugated secondary antibody diluted in Can Get Signal Immunoreaction Enhancer Solution 2 (TOYOBO) at room temperature (RT) for 1\u00a0h. After the membrane was washed with TBS-T, it was developed with Chemi-Lumi One L or Super . The intensity of each western blot signal was quantified by Image J software ("} +{"text": "I with the unsymmetrical ligand N-(pyridine-2-ylmeth\u00adyl)pyridine-3-amine afforded right- and left-handed helical chains. The AgI atom of the right-handed helical chain adopts a slightly distorted linear coordination geometry, while that of the left-handed helical chain displays a bent geometry.The reaction of Ag 11H11N3)]CF3SO3}n, there are two AgI atoms, two N-(pyridine-2-ylmeth\u00adyl)pyridine-3-amine ligands (A and B) and two CF3SO3\u2212 anions. Both AgI atoms are bridged by two pyridine N atoms from two symmetry-related A or B ligands, forming right- or left-handed helical chains, respectively. The AgI atom of the right-handed helical chain adopts a slightly distorted linear coordination geometry [N\u2014Ag\u2014N = 170.69\u2005(14)\u00b0], while that of the left-handed helical chain adopts a bent geometry [N\u2014Ag\u2014N = 149.42\u2005(14)\u00b0]. Both helical chains have the same pitch length [10.8437\u2005(5)\u2005\u00c5], propagate along the b-axial direction and are alternately arranged via Ag\u22efAg [3.0814\u2005(5)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.514\u2005(3) and 3.487\u2005(3)\u2005\u00c5], resulting in the formation of a two-dimensional supra\u00admolecular network extending parallel to the ab plane. Weak Ag\u22efO and Ag\u22efF [3.017\u2005(3)\u2005\u00c5] inter\u00adactions as well as N\u2014H\u22efO and C\u2014H\u22efO, C\u2014H\u22efN and C\u2014H\u22efF hydrogen-bonding inter\u00adactions occur between the helical chains and the anions.In the asymmetric unit of the title compound, {[Ag(C The structure of title compound is related to that of the perchlorate salt , two N-(pyridine-2-ylmeth\u00adyl)pyridine-3-amine and two tri\u00adfluoro\u00admethane\u00adsulfonate anions. The Ag1 atom is coordinated by two pyridine N atoms from two symmetry-related A ligands giving a geometry which is slightly distorted from linear [N1\u2014Ag1\u2014N2 = 170.69\u2005(14)\u00b0], forming a right-handed helical chain, while the Ag2 atom is coordinated by two pyridine N atoms from two symmetry-related B ligands in a bent arrangement [N4\u2014Ag2\u2014N5 = 149.42\u2005(14)\u00b0], forming a left-handed helical chain. Two pyridine rings coordinating to the Ag1 and Ag2 atoms are tilted by 14.1\u2005(3) and 28.9\u2005(2)\u00b0, respectively, with respect to each other.The molecular components of the title structure are shown in Fig.\u00a01b-axial direction and are alternately arranged via Ag1\u22efAg2 inter\u00adactions [3.0814\u2005(5)\u2005\u00c5], resulting in the formation of a two-dimensional supra\u00admolecular network extending parallel to the ab plane \u2005\u00c5; symmetry code: (iv) \u2212x, y\u00a0\u2212\u00a0z\u00a0+\u00a03SO3\u2212 anions.Both helical chains in the structure have the same pitch length [10.8437\u2005(5)\u2005\u00c5], propagate along the ne Fig.\u00a02. Further] Figs. 1 and 2 \u25b6 N-(pyridin-2-ylmeth\u00adyl)pyridine-3-amine) was prepared according to a procedure described by Lee et al. = 0.95\u2005\u00c5 for Csp2\u2014H, 0.88\u2005\u00c5 for amine N\u2014H and 0.99\u2005\u00c5 for methyl\u00adene C\u2014H. For all H atoms Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814022922/zs2318sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S1600536814022922/zs2318Isup2.hklStructure factors: contains datablock(s) I. DOI: 1029928CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A near-perfect octa\u00adhedral zinc(II) complex coordinated to two coumarin fluoro\u00adphores. 14H14NO4)2(H2O)2]\u00b72C2H6OS, shows that the ZnII cation adopts an octa\u00adhedral geometry and lies on an inversion center. Two organic ligands occupy the equatorial positions of the coordination sphere, forming a chelate ring motif via the O atom on the formyl group and another O atom of the carbonyl group (a pseudo-\u03b2-diketone motif). Two water mol\u00adecules occupy the remaining coordination sites of the ZnII cation in the axial positions. The water mol\u00adecules are each hydrogen bonded to a single dimethyl sulfoxide mol\u00adecule that has been entrapped in the crystal lattice. The structure of the title coordination complex, [Zn(C This is indicated by the short C=O bond of the dione (O3\u2014C4) and the C=O bond length of the formyl moiety (O4\u2014C9), with values of 1.2686\u2005(10) and 1.2603\u2005(10)\u2005\u00c5, respectively. The Zn\u2014O bonds complete the stable six-membered chelating motif, which is favorable for smaller metal ions distance is 2.09\u2005\u00c5 to 93.18\u2005(3)\u00b0. A single DMSO solvent mol\u00adecule completes the asymmetric unit.The mol\u00adecular structure of (1) is shown in Fig.\u00a01II ion S1\u2014O6\u22efH52\u2014O5 [1.983\u2005(9)\u2005\u00c5]. Inter\u00adestingly, there are also two C\u2014H\u22efO hydrogen-bonding inter\u00adactions from the methyl moiety of DMSO; one with the O atom on the formyl functional group in the equatorial position (H13A\u22efO4 = 2.52\u2005\u00c5) and an additional hydrogen-bonding inter\u00adaction from the carbonyl\u00addione group occupying another equatorial position (H12B\u22efO3 = 2.62\u2005\u00c5). Together these two inter\u00adactions form three C\u22efO6 (2.29\u2005\u00c5), forming an infinite chain.The crystal structure of the title compound shows an extensive array of hydrogen-bonding inter\u00adactions Table\u00a01 forming et al., 2013A) and the electron-rich region of the second coumarin ring system (C4A\u2013C8A) of an adjacent compound, whereby the centroids are 3.734\u2005\u00c5 apart and 5(b); however, these did not yield any results. Therefore a modification of the search by specifically searching structures that have a bidentate chelating \u03b2-diketone motif coordinated to the zinc(II) in the equatorial position, with two water mol\u00adecules in the axial position, as shown in Fig.\u00a05c) was carried out. This refined search yielded two similar structures with ZnII octahedrally coordinated, the first by Solans et al., whereby two 1,3-bis\u00ad(2-hy\u00addroxy\u00adphen\u00adyl)propane-1,3-dionate ligands coordinate to the ZnII ion, with the remaining two coordination sites occupied by two ethanol mol\u00adecules was then added to the solution. After stirring for 20\u2005min, a yellow solid formed, which was collected by filtration and dried. A small amount of the solid (20\u2005mg) was redissolved in a 1:1 mixture of MeOH and DMSO to form a saturated solution (1\u2005mL) which was was allowed to stand for several weeks to form the title compound as colorless needles suitable for X-ray analysis. 1H NMR : \u03b4 9.68 , 7.91 , 6.53 , 6.33 , 3.41 , 1.23 ; 13C NMR \u03b4 192.2, 169.1, 165.8, 159.5, 157.7, 153.3, 128.3, 108.4, 108.0, 102.8, 96.9, 44.9, 40.6, 29.7, 12.5; LRMS\u2013ESI (negative mode), NaCl was added as a charging agent [M \u2212 2H2O + Cl]\u2212 = 619 m/z, [M \u2212 H2O \u2212 C14H15NO4 + 2Cl]\u2212 = 396 m/z, CID 396 yields [C14H15NO4]\u2212 = 261 m/z; IR (ATR solid); 3364 \u03bdOH, 2972, 2926 (m) \u03bdCH, 1722 (m) \u03bdCO (\u03b4-lactone), 1689 \u03bdCO (ketone), 1590 \u03bdCO (form\u00adyl), 564 \u03bdCO (Zn\u2014O) cm\u22121.7-(Di\u00adethyl\u00adamino)-4-hy\u00addroxy\u00adcoumarin was dissolved in 2-propanol (20\u2005mL), tri\u00adethyl \u00adorthoformate and 2-amino\u00adpyriimidine were added and the solution was heated to reflux for 4\u2005h. Upon cooling, the solid was collected and used without further purification. This compound was then dissolved in methanol (10\u2005mL), to which Zn(OAc)sp2, 0.99\u2005\u00c5 for CH2, and 0.98\u2005\u00c5 for methyl groups. Those on O atoms were assigned from difference maps, and their positions refined, with O\u2014H distances restrained to 0.86\u2005(1)\u2005\u00c5. Uiso values for H atoms were assigned as 1.2 times Ueq of the attached atoms (1.5 for methyl and water groups).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016009853/zl2668sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016009853/zl2668Isup2.hklStructure factors: contains datablock(s) I. DOI: 1486125CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal of the title purine derivative, mol\u00adecules are linked by O\u2014H\u22efN, N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, forming a three-dimensional framework. 7H7N5O3, is an adduct of guanine with glyoxal. In the mol\u00adecule, the di\u00adhydro\u00adimidazole ring adopts a twisted conformation on the C\u2014C bond, and the two hydroxyl groups lie on opposite sides of the mean plane of the ring. In the crystal, the mol\u00adecules are linked by N\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efN hydrogen bonds forming a three-dimensional framework. The crystal packing is reinforced by C\u2014H\u22efO hydrogen bonds and by offset \u03c0\u2013\u03c0 stacking of the purine ring systems of inversion related mol\u00adecules [inter\u00adcentroid distance = 3.4839\u2005(12)\u2005\u00c5].The title purine derivative, C Purine derivatives have been developed as inhibitors of cyclin-dependent kinase \u2005\u00c5, inter\u00adplanar distance = 3.311\u2005(1)\u2005\u00c5, slippage = 1.112\u2005\u00c5; Cg2 and Cg3 are the centroids of the N3/C4/C3/N4/C5 and N1/C1/C4/C3/N2/C2 rings, respectively; symmetry code: (i) \u2212x, \u2212y, \u2212z\u00a0+\u00a02].In the crystal, mol\u00adecules are also linked e Table\u00a01. The laye Table\u00a01. Within e Table\u00a01, and invet al., 2016H-6-one as substructure, gave 61 hits. Many of these compounds concern guanine and guaninium and some metal complexes, but none involve a fused third ring. The structure of the title compound has not been reported previously.A search of the Cambridge Structural Database medium.The title compound was synthesized according to a literature method = 1.2Ueq(C) and 1.5Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016009087/xu5887sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016009087/xu5887Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016009087/xu5887Isup3.cmlSupporting information file. DOI: 1469976CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III compounds with 4-amino\u00adbenzoic acid and 4-chloro-3-nitro\u00adbenzoic acid, discrete centrosymmetric bridged dinuclear complex units are present giving an overall three-dimensional hydrogen-bonded structure in the first complex and a one-dimensional coordination polymer in the second.In the structures of two Er 2-4-amino\u00adbenzoato-\u03ba2O:O\u2032)bis\u00ad[bisdi\u00adaqua\u00aderbium(III)] dihydrate, [Er2(C7H6NO2)6(H2O)4]\u00b72H2O, (I), and 4-chloro-3-nitro\u00adbenzoic acid (CLNBAH), namely poly[hexa\u00adkis\u00ad(\u03bc2-4-chloro-3-nitro\u00adbenzoato-\u03ba2O:O\u2032)bis\u00ad(dimethyl sulfoxide-\u03baO)dierbium(III)], [Er2(C7H3ClNO4)6(C2H6OS)2]n, (II), have been determined. In the structure of solvatomorphic compound (I), the symmetry-related irregular ErO8 coordination polyhedra in the discrete centrosymmetric dinuclear complex comprise two monodentate water mol\u00adecules and six carboxyl\u00adate O-atom donors, four from two bidentate carboxyl\u00adate O,O\u2032-chelate groups and two from the bis-monodentate O:O\u2032-bridging group of the third 4-ABA anion. The Er\u2014O bond-length range is 2.232\u2005(3)\u20132.478\u2005(3)\u2005\u00c5 and the Er\u22efEr separation in the dinuclear complex unit is 4.7527\u2005(4)\u2005\u00c5. One of the coordinating water mol\u00adecules is involved in an intra-unit O\u2014H\u22efO hydrogen-bonding association with an inversion-related carboxyl\u00adate O-atom acceptor. In contrast, the anhydrous compound (II) is polymeric, based on centrosymmetric dinuclear repeat units comprising ErO7 coordination polyhedra which involve four O-atom donors from two bidentate O:O\u2032-bridging carboxyl\u00adate groups, one O-atom donor from the monodentate dimethyl sulfoxide ligand and two O-atom donors from the third bridging CLNBA anion. The latter provides the inter-unit link in the one-dimensional coordination polymer extending along [100]. The Er\u2014O bond-length range in (II) is 2.239\u2005(6)\u20132.348\u2005(6)\u2005\u00c5 and the Er\u22efEr separation within the dinuclear unit is 4.4620\u2005(6)\u2005\u00c5. In the crystal of (I), extensive inter-dimer O\u2014H\u22efO and N\u2014H\u22efO hydrogen-bonding inter\u00adactions involving both the coordinating water mol\u00adecules and the solvent water mol\u00adecules, as well as the amine groups of the 4-ABA anions, give an overall three-dimensional network structure. Within this structure are also weak \u03c0\u2013\u03c0 ring inter\u00adactions between two of the coordinating ligands [ring-centroid separations = 3.676\u2005(3) and 3.711\u2005(2)\u2005\u00c5]. With (II), only weak intra-polymer C\u2014H\u22efO, C\u2014H\u22efCl and C\u2014H\u22efS inter\u00adactions are present.The crystal structures of two erbium(III) complexes with 4-amino\u00adbenzoic acid (4-ABAH), namely bis\u00ad(\u03bc RE) metals has been investigated extensively and the structures of a large number of complexes with various ligand types are known series of complexes (La\u2013Tb as well as Dy and Er), in which the structures are two-dimensional, the second triclinic (P2(4-ABA)6(H2O)2]; triclinic [[Tb2(4-ABA)6(H2O)2]\u00b72H2O]} is of inter\u00adest and its occurrence was indicated as being dependent on pH control in the preparation.The coordination chemistry of the rare earth (RE series (predominantly triclinic) might also show the same effect so this was tested with Er in a reaction of erbium(III) acetate with 4-ABA in aqueous ethanol under mild reaction conditions, with no additional pH control. The title triclinic complex [Er2(C7H6NO2)6(H2O)4]\u00b72H2O, (I)3+ complex with 4-ABA , b = 11.0117\u2005(1), c = 12.7430\u2005(2)\u2005\u00c5, \u03b1 = 89.372\u2005(5), \u03b2 = 72.0360\u2005(6), \u03b3 = 75.0730\u2005(7)\u00b0, V = 1169.97\u2005(2)\u2005\u00c53, confirming that the two are isotypic.It was considered that some of the other later members of the 2(C7H3ClNO4)6(C2H6OS)2]n, was obtained in a similar reaction to (I)Complex (II)8 complex units , four O-atom donors from two slightly asymmetric bidentate O,O\u2019 chelate carboxyl\u00adate groups (the A and B 4-ABA ligands) and two bridging O-atom donors from two symmetry-related ligands (C). The Er\u22efEri separation in the dinuclear unit is 4.7527\u2005(4)\u2005\u00c5. Unlike the polymeric solvatomorphic ErIII complex [Er2(4-ABA)6(H2O)2]n\u00b7nH2O carboxyl\u00adate hydrogen bond is present between one of the the coordinating water mol\u00adecules (O1W) and an inversion-related carboxyl\u00adate O-atom (O11Ai) ] Table\u00a01, compris) Table\u00a02. This stO,O\u2032-chelate ligands (A) and the bridging ligand (C), the groups are essentially coplanar with the benzene ring , while in the second bidentate chelate ligand (B) the group is rotated out of the plane [corresponding torsion angle = 155.9\u2005(4)\u00b0]. Such a \u2019planar\u2019 conformation is also found in the structure of the parent acid III atoms , a monodentate DMSO O-atom and O-donors (O12Ci) and O11Ci from the C ligand which extends the dinuclear unit into a one-dimensional coordination polymer lying along [100] \u2005\u00c5. Also present within the repeat unit are a C2B\u2014H\u22efO11 hydrogen bond [3.298\u2005(13)\u2005\u00c5] and a C2A\u2014H\u22efS1 inter\u00adaction [3.743\u2005(10)\u2005\u00c5] ms Fig.\u00a02 being se0] Fig.\u00a03. The Er\u20140] Fig.\u00a03 and the ] Table\u00a04.A/B/C\u2014C1A/B/C\u2014C11A/B/C\u2014O11A/B/C = 158.7\u2005(9), 177.2\u2005(9) and 160.3\u2005(8)\u00b0, respectively. The torsion angles of the nitro groups C2A/B/C\u2014C3A/B/C\u2014N3A/B/C\u2014O32A/B/C are \u2212150.4\u2005(12), 174.1\u2005(16) and 120.3\u2005(13)\u00b0, respectively. In the structure of the parent CLNBAH acid water and Ocarbox\u00adyl hydrogen bonds give a three-dimensional network structure \u2005\u00c5] and C ligands [C\u22efCviii = 3.676\u2005(3)\u2005\u00c5] 6(H2O)4]2\u00b76H2O e Figs. 4 and 5 \u25b8.With (II)The title compounds were synthesized by warming together for 10\u2005min, a solution obtained by mixing 5\u2005ml of ethano\u00adlic 4-amino\u00adbenzoic acid (1\u2005mmol: 135\u2005mg) [for (I)] or 4-chloro-3-nitro\u00adbenzoic acid (1\u2005mmol: 200\u2005mg) [for (II)], with 10\u2005ml of aqueous erbium(III) acetate hexa\u00adhydrate (0.3\u2005mmol: 216\u2005mg). Partial room-temperature evaporation of these solutions provided pale-pink block-like single crystals of (I)Uiso(H) = 1.5Ueq(O) or 1.2Ueq(N). Other H atoms were included in the refinement at calculated positions , using a riding-model approximation. In the refinement of (II)\u22123) located within 1.0\u2005\u00c5 of the Er1 site were present. These are possibly due to poor crystal quality coupled to effects of an insufficient absorption correction.Crystal data, data collection and structure refinements for (I)10.1107/S2056989015020319/wm5228sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989015020319/wm5228Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989015020319/wm5228IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1433543, 1433542CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-[3-(di\u00admethyl\u00adamino)\u00adprop\u00adyl]-N\u2032-(2-hy\u00addroxy\u00adphen\u00adyl)oxamide trianion bridges two CuII cations to form the binuclear complex, in which the CuII cations have distorted square-planar and square-pyramidal coordination geometries.The 2(C13H16N3O3)(C12H6N2O2)(H2O)]ClO4\u00b70.5H2O, consists of a cis-oxamide-bridged binuclear CuII complex cation, a perchlorate anion and half a solvent water mol\u00adecule. One CuII cation is N,N\u2032,N\",O-chelated by an N-[3-(di\u00admethyl\u00adamino)\u00adprop\u00adyl]-N\u2032-(2-hy\u00addroxy\u00adphen\u00adyl)oxamide trianion in a distorted square-planar geometry, whereas the other CuII cation is O,O\u2032-chelated by the oxamide moiety of the anion and N,N\u2032-chelated by a 1,10-phenanthroline-5,6-dione mol\u00adecule, and a water mol\u00adecule further coordinates the second CuII cation, completing a distorted square-pyramidal coordination geometry. In the crystal, classical O\u2014H\u22efO hydrogen bonds, weak C\u2014H\u22efO hydrogen-bonding inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions link the complex cations, anions and solvent water mol\u00adecules into a three-dimensional supra\u00admolecular architecture. In the crystal, the di\u00admethyl\u00adamino\u00adpropyl unit of the oxamide anion is disordered over two positions with an occupancy ratio of 0.561\u2005(11):0.439\u2005(11); the solvent water mol\u00adecule is also disordered over two positions, the occupancy ratio being 0.207\u2005(10):0.293\u2005(10).The title compound, [Cu Its crystal structure and supra\u00admolecular structure are reported here. 1,10-Phenanthroline-5,6-dione (Phdo) is a multifaceted ligand since the structure and electronic properties thereof incorporate the features of the di\u00adimine and quinone functionalities \u2005\u00c5, which is consistent with structures reported previously \u2005\u00c5 for N4], and a water mol\u00adecule (O4) occupies the apical position, completing a distorted square-pyramidal coordination geometry with a \u03c4 value of 0.06 cations with three planar five-membered chelate rings and one six-membered ring, the latter being disordered over two positions. The puckering parameters of the first component (containing atoms C10A and C11A) are Q = 0.554\u2005(8)\u2005\u00c5, \u03b8 = 47.6\u2005(6)\u00b0 and \u03d5 = 206.0\u2005(7)\u00b0, and those of the other are Q = 0.565\u2005(11)\u2005\u00c5, \u03b8 = 123.4\u2005(8)\u00b0 and \u03d5 = 38.8\u2005(9)\u00b0; both suggest an approximate half-chair conformation.The hexa\u00addentate oxamide anion, Dmapoxx, 1\u00a0\u2212\u00a0y, \u2212z [symmetry code (iv)], and vice versa , 3.211\u2005(4) (C3iv) and 3.252\u2005(4)\u2005\u00c5 (C19iv).Besides classical O\u2014H\u22efO hydrogen bonds, weak C\u2014H\u22efO hydrogen bonds and aromatic stacking inter\u00adactions are important to the supra\u00admolecular structure. As illustrated in Fig.\u00a02sa Fig.\u00a03. The sepII complexes of 1,10-phenanthroline-5,6-dione have been reported previously, for example, Chetana et al. \u00adprop\u00adyl]-N\u2032-2-(oxidophen\u00adyl)oxamide 2\u00b76H2O in methanol (5\u2005ml) was added dropwise to a solution of H3Dmapox and piperidine in methanol (5\u2005ml). The solution was stirred continuously for 0.5\u2005h. Then a solution of Phdo in methanol (5\u2005ml) was added dropwise, and the mixture was stirred continuously at 313\u2005K for 6\u2005h and then filtered. Dark-blue crystals of the title compound suitable for X-ray analysis were obtained from the filtrate by slow evaporation at room temperature for 7\u2005d. Yield: 0.026\u2005g (71.62%). Analysis calculated for Cu2C25H25ClN5O10.5: C 41.44, H 3.48, N 9.67%; found: C 42.57, H 3.15, N 9.19%.A\u2013C13A, with occupancies of 0.561\u2005(11); C10B\u2013C13B, 0.439\u2005(11)], three oxygen atoms of the perchlorate ion and the solvent water mol\u00adecule ; O7B, 0.293\u2005(10)]. The occupancies were refined freely except for the sum of atoms O7A and O7B which was fixed at 0.5. Some restraints on distances (DFIX) and anisotropic displacement parameters (SIMU) were applied to the disordered atoms to avoid unreasonable geometries. The hydrogen atoms of the water mol\u00adecules were found in a difference Fourier map and then refined as riding. Other H atoms were placed in calculated positions, with C\u2014H = 0.96 (meth\u00adyl), 0.97 (methyl\u00adene) and 0.93\u2005\u00c5 (aromatic), and refined using a riding model, with Uiso(H) = 1.2 Ueq(C) or 1.5 for methyl groups.Crystal data, data collection, and refinement details are summarized in Table\u00a0310.1107/S2056989015009391/xu5850sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015009391/xu5850Isup2.hklStructure factors: contains datablock(s) I. DOI: 1401557CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The latter is coordinated by two Cl atoms and two N,N\u2032-di\u00admethyl\u00adthio\u00adurea (Dmtu) ligands through their S atoms, defining a distorted tetra\u00adhedral coordination sphere with bond angles in the range 102.47\u2005(4)\u2013118.32\u2005(4)\u00b0. Intra- and inter\u00admolecular hydrogen bonds of the type N\u2014H\u22efCl with S(6) and R22(12) ring motifs are present. The inter\u00admolecular contacts make up polymeric chains extending parallel to [101].The mol\u00adecular structure of the title compound, [HgCl L) or derivatives thereof have shown that in combination with a halide or pseudohalide X, some of the complexes exist as mononuclear species [HgX2L2] 2 complexes with methyl\u00adthio\u00adurea as an auxiliary ligand complexes with thio\u00adurea ligands (2 with di\u00admethyl\u00adthio\u00adurea (Dmtu) as an additional ligand, [HgCl2(C3H8N2S)2], (I)In this article, we report on synthesis and crystal structure of HgCl2L2] complexes et al., 1995S(6) loop motifs 2] 2] 2] 2Cl2] and [Hg(tetra\u00admethyl\u00adthio\u00adurea)2Cl2] 2] and [CdBr2(Dmtu)2], the coordination spheres around Cd deviate only slightly from ideal tetra\u00adhedral values. On the other hand in [Hg(Dmtu)2(CN)2], the HgII atom exhibits a severely distorted tetra\u00adhedral coordination sphere with bond angles in the range 94.31\u2005(2) to 148.83\u2005(13)\u00b0 HgClUiso(H) = 1.5Ueq(C) and Uiso(H) = 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015015406/wm5202sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015015406/wm5202Isup2.hklStructure factors: contains datablock(s) I. DOI: 1419298CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In both dimers, the platinum(II) complexes are arranged anti\u00adparallel to each other. Each complex exhibits a slightly distorted square-planar coordination environment around the central Pt(II) atom. The dihedral angles between two chelate rings including the PtII atom in these complexes are 0.08\u2005(12) and 1.54\u2005(9)\u00b0.The title cyclo\u00admetalated platinum(II) complex with 2-(4-bromo\u00adphen\u00adyl)pyridinato and acetyl\u00adacetonato ligands, [Pt(C Although these complexes afford luminescence in the solid state, their crystal structures have not been sufficiently explored. We report herein the crystal structure of the cyclo\u00admetalated platinum(II) complex with 2-(4-bromo\u00adphen\u00adyl)pyridinato and acetyl\u00adacetonato ligands, [Pt(Brppy)(acac)].Square-planar cyclo\u00admetalated platinum(II) complexes with luminescent properties have recently attracted attention because of their potential applications are slightly distorted from an ideal square-planar configuration, with angles around Pt1 in the range 81.89\u2005(18)\u201393.04\u2005(17)\u00b0 and around Pt2 in the range 81.73\u2005(18)\u201393.57\u2005(16)\u00b0. The Pt\u2014C bond lengths [Pt1\u2014C11 = 1.970\u2005(5) and Pt2\u2014C27 = 1.969\u2005(5)\u2005\u00c5] are slightly shorter than the Pt\u2014N bond lengths [Pt1\u2014N1 = 1.995\u2005(4) and Pt2\u2014N2 = 1.999\u2005(4)\u2005\u00c5] due to the stronger electron-donating ability of a C atom compared to that of an N atom. Pt\u2014O bond lengths are compiled in Table\u00a01The asymmetric unit of the title compound contains two complex mol\u00adecules with very similar configurations (r.m.s. deviation of fit of two molecules = 0.07\u2005\u00c5). The structure of one of the complex mol\u00adecules of the title compound is shown in Fig.\u00a01II complex (Pt1) is directed to the b axis, on the other hand, that of the other complex (Pt2) is directed to the a axis. The shortest inter\u00admolecular contacts are C4\u22efC15i = 3.406\u2005(7) and C22\u22efO3ii = 3.402\u2005(6)\u2005\u00c5 . Weak C\u2014H\u22efO and C\u2014H\u22efBr inter\u00adactions might also help to consolidate the crystal packing and Pt2\u22efPt2ii = 3.723\u2005(1)\u2005\u00c5] are longer than the van der Waals diameter of the Pt atom (\u03bc-Cl)]2 ([Pt(C\u03bc2:-Cl)] A mixture of 2-(4-bromo\u00adphen\u00adyl)pyridine and K2PtCl4 in a 2-eth\u00adoxy\u00adethanol\u2013water mixture (45\u2005ml/15\u2005ml) was stirred for 6\u2005h at 333\u2005K under an Ar atmosphere. After cooling to room temperature, the yellow\u2013green precipitate was filtered off, washed with di\u00adchloro\u00admethane, and dried in vacuo. Yield: 0.535\u2005g, (48.2%).11H7BrN)(C5H7O2)]:: 262\u2005(29800), 280\u2005(27500), 317 , 330 , 363\u2005(6400), 389\u2005(4200). 1H NMR ; 8.97 , 7.81 , 7.71 , 7.57 , 7.31-7.45 , 7.14 , 5.48 , 2.03 , 2.01 .Analysis found = 1.2Ueq(C) for Csp2\u2013H, and Uiso(H) = 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015017478/wm5214sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015017478/wm5214Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015017478/wm5214Isup3.tifSupporting information file. DOI: 1425736CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-(11-hy\u00addroxy\u00adundec\u00adyl)isoxazole-3-carboxamide hemihydrate, a derivative of anti\u00adviral \u2018WIN compounds\u2019, are reported.The crystal structure and supra\u00admolecular features of 5-{3-prop\u00adyl}- 29H42N4O5\u00b70.5H2O, comprises four structural units. A flexible prop\u00adyloxy unit in a gauche conformation, with a \u2013C(H2)\u2014C(H2)\u2014C(H2)\u2014O\u2013 torsion angle of \u221264.32\u2005(18)\u00b0, connects an isoxazole ring and an approximately planar phenyl\u00adoxa\u00addiazole ring system [with a maxixmum devation of 0.061\u2005(2)\u2005\u00c5], which are oriented almost parallel to one another with a dihedral angle of 10.75\u2005(7)\u00b0. Furthermore, a C11-alkyl chain with a terminal hy\u00addroxy group links to the 3-position of the isoxazole ring via an amide bond. In the crystal, a half-occupancy solvent water mol\u00adecule connects to a neighbouring mol\u00adecule via an inter\u00admolecular O\u2014H\u22efO(water) hydrogen bond to the C11-alkyl chain hy\u00addroxy group.The title compound, C The three heterocyclic rings are approximately coplanar to one another, having dihedral angles between the rings of 11.57\u2005(8) (C19\u2013C21/N22/O23 and C7\u2013C12), 10.68\u2005(9) (C19\u2013C21/N22/O23 and C2/O3/N4/C5/N6) and 4.81\u2005(9)\u00b0 (C7\u2013C12 and C2/O3/N4/C5/N6), maintaining the WIN framework in a linear conformation. The dihedral angle between the isoxazole ring (C19\u2013C21/N22/O23) and the approximately planar phenyl\u00adoxa\u00addiazole ring system is 10.75\u2005(7)\u00b0. The isoxazole and phenyl\u00adoxa\u00addiazole ring systems are connected by a prop\u00adyloxy unit (O15\u2013C18), which is in a gauche conformation, with a C18\u2014C17\u2014C16\u2014O15 torsion angle of \u221264.32\u2005(18)\u00b0. The amide group (N26\u2013C24) at the 3-position of the isoxazole ring which joins the C11-alkyl chain (C27\u2013O38) and the WIN framework is likewise almost coplanar with the isoxazole ring, with a dihedral angle of 10.92\u2005(9)\u00b0 between the amide (H26/N26/C24/O25) and isoxazole planes. The amide hydrogen (H26) and the acidic isoxazole hydrogen (H20) are on opposite sides, with a torsion angle (N26\u2014C24\u2014C21\u2014C20) of 172.31\u2005(15)\u00b0. The C11-alkyl chain (C27\u2013C37) is in an all-anti conformation, with an average torsion angle of 178.80\u00b0. The WIN framework and the C11-linker arm structural units are aligned roughly in a 160\u00b0 angle and the total length of the title mol\u00adecule measures up to 3.4\u2005nm.The mol\u00adecular structure of the title compound is shown in Fig.\u00a0111-alkyl chain hy\u00addroxy [O\u2014H\u22efO = 1.90\u2005(1)\u2005\u00c5], solvent water [O\u2014H\u22efO = 1.87\u2005(1)\u2005\u00c5], amide carbonyl and isoxazole hydrogen (C\u2014H\u22efO = 2.56\u2005\u00c5) groups of two parallel neighbouring mol\u00adecules and The title compound packs in the crystal lattice in layers, in which the mol\u00adecules are held together by solvent-mediated O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds (motif 1), as well as C\u2014H\u22efN and C\u2014H\u22efO inter\u00admolecular inter\u00adactions between the heterocyclic isoxazole and phenyl\u00adoxa\u00addiazole units of neighbouring mol\u00adecules (motif 2) Table\u00a01. In the es Fig.\u00a02. In a sips Fig.\u00a02.et al., 2014et al., 2014et al., 2014et al., 2004et al., 2014et al., 2004et al., 2014et al., 2014ca 60.8\u00b0. In all of the structures, the prop\u00adyloxy unit is in a gauche conformation, with torsion angles in the range 62.4\u201369.2\u00b0.A search of the Cambridge Structural Database \u00adprop\u00adyl]-N-ethyl\u00adcarbodi\u00adimide (0.19\u2005mmol) and a catalytic amount of 1-hy\u00addroxy\u00adbenzotriazole at 273\u2005K gave the title compound in 68% yield after subsequent chromatographic purification in silica with a di\u00adchloro\u00admethane\u2013methanol mixture (95:5 v/v). Needle-like crystals of the title compound were obtained from an ethanol solution by vapor diffusion with water.An amide coupling reaction of 5-{3-prop\u00adyl}isoxazole-3-carb\u00adoxy\u00adlic acid = 1.5Ueq(C) for methyl and 1.2Ueq(C) for other H atoms, and N\u2014H = 0.88\u2005\u00c5 and Uiso(H) = 1.2Ueq(N). The positions of the O-bound H atoms were located in a difference Fourier map and refined as riding atoms with Uiso(H) = 1.5Ueq(O). The O\u2014H distance of the half-occupied water molecule was restrained to 0.84\u2005(1)\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015007367/lh5758sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015007367/lh5758Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015007367/lh5758Isup3.cmlSupporting information file. DOI: 1059505CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information: interactive version of Fig. 3Enhanced figure:"} +{"text": "The basal plane consists of two N atoms [Cu\u2014N = 2.008\u2005(3) and 2.032\u2005(3)\u2005\u00c5] from the phenanthroline ligand, and of two carboxyl\u00adate O atoms [Cu\u2014O = 1.942\u2005(3) and 1.948\u2005(3)\u2005\u00c5] from two 2,3,4,5-tetra\u00adfluoro\u00adbenzoate anions. Another 2,3,4,5-tetra\u00adfluoro\u00adbenzoate anion provides the apical carboxyl\u00adate O atom [Cu\u2014O = 2.262\u2005(3)\u2005\u00c5] and bridges two CuII ions into a binuclear centrosymmetric dimer. Intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between one of the tetra\u00adfluoro\u00adbenzene rings and the middle of the phenenanthroline rings [3.617\u2005(3)\u2005\u00c5] stabilize the mol\u00adecular configuration. O\u2014H\u22efO hydrogen bonds between the lattice water mol\u00adecules and the unbound carboxyl\u00adate O atoms of the metal complexes leads to the formation of a chain structure parallel to [100].In the title compound, [Cu DOI: 10.1107/S1600536814022065/wm5062Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814022065/wm5062fig1.tifx y z . DOI: x\u00a0+\u00a01, \u2013y\u00a0+\u00a01, \u2013z\u00a0+\u00a02.The mol\u00adecular structure of the title compound, with displacement ellipsoids drawn at the 30% probability level for non-H atoms. The non-labelled atoms are generated by symmetry code \u2013Click here for additional data file.10.1107/S1600536814022065/wm5062fig2.tif. DOI: The packing of the mol\u00adecular entities of the title compound. O\u2014H\u22efO hydrogen-bonding inter\u00adactions are indicated by dashed lines.1027857CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II ion atom is octa\u00adhedrally coordinated in di\u00adaqua\u00adtris\u00ad(1-ethyl-1H-imidazole)\u00adsulfato\u00adnickel(II). There are three organic ligands, two water and the sulfate anion coordinated around the NiII centre. Two complex mol\u00adecules form an inversion dimer via two pairs of O\u2014H\u22efO hydrogen bonds between the coordinating sulfate anion and a water mol\u00adecule in the unit cell.The Ni 4)(C5H8N2)3(H2O)2], the NiII ion is coordinated by three facial 1-ethyl-1H-imidazole ligands, one monodentate sulfate ligand and two water mol\u00adecules in a slightly distorted octa\u00adhedral coordination environment. In the crystal, two pairs of O\u2014H\u22efO hydrogen bonds link complex mol\u00adecules, forming inversion dimers incorporating R24(8), R22(8) and R22(12) rings. The dimeric unit also contains two symmetry-unique intra\u00admolecular O\u2014H\u22efO hydrogen bonds. In addition, weak C\u2014H\u22efO hydrogen bonds, weak C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 inter\u00adactions with a centroid\u2013centroid distance of 3.560\u2005(2)\u2005\u00c5 combine to form a three-dimensional network. One of the ethyl groups is disordered over two sets of sites with occupancies in the ratio 0.586\u2005(7):0.414\u2005(7).In the title complex, [Ni(SO H-imidazole mol\u00adecules as ligands. The title compound was prepared by the reaction of NiSO4\u00b76H2O and 1-ethyl-1H-imidazole. The crystal structure of the title compound is presented herein.In spite of efforts in the past decades to synthesize structurally highly varying metal-organic complexes, no structures up to this point have been reported which contain the combination of a hydro\u00adphilic sulfate anion, water mol\u00adecules and hydro\u00adphobic 1-ethyl-1II ion is coordinated in a slightly distorted octa\u00adhedral geometry by three facially arranged 1-ethyl-1H-imidazole ligands, one monodentate sulfate ligand and two water mol\u00adecules. The Ni\u2014N bond lengths are in the range 2.0630\u2005(16)\u20132.0817\u2005(17)\u00c5 and the Ni\u2014O bond lengths are in the range 2.1195\u2005(15)\u20132.1502\u2005(14). The Niii ion is displaced by 0.1038\u2005(3)\u2005\u00c5 from the O1/O2/N11/N13 plane. The distances of two water O atoms O1 and O2 from the S1/O3/Ni1/N12 plane are the same within experimental error, with values of 1.520\u2005(2) and \u22121.504\u2005(2)\u2005\u00c5, respectively. The sulfate atom O6 is displaced by only 0.144\u2005(2)\u2005\u00c5 from the S1/O3/Ni1/N12 plane, while atoms O4 and O5 are displaced by 1.114\u2005(2) and \u22121.298\u2005(2)\u2005\u00c5, respectively, from this plane .In the crystal, two pairs of O\u2014H\u22efO hydrogen bonds Table\u00a01 link comds Fig.\u00a03. In addiet al., 2004et al., 2003et al., 2009et al., 2006et al., 1995et al., 2011et al., 2002et al., 2000A search of the Cambridge Structural Database I. DOI: 10.1107/S2056989016002863/lh5802Isup2.hklStructure factors: contains datablock(s) I. DOI: 1454040CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the complex cation, two MnII atoms are bridged by two O atoms of two different 2-benzoyl\u00adbenzoate ligands, each MnII atom being further coordinated by two 2,2\u2032-bi\u00adpyridine (bipy) ligands in a distorted octa\u00adhedral environment. Within the binuclear mol\u00adecule, the Mn\u22efMn separation is 4.513\u2005(7)\u2005\u00c5. Inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef \u03c0 inter\u00adactions link the mol\u00adecules into a three-dimensional network. The title compound, [Mn DOI: 10.1107/S2056989015023671/bg2577Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015023671/bg2577fig1.tifx y z . DOI: x\u00a0+\u00a01, \u2212y, \u2212z\u00a0+\u00a02.The mol\u00adecular structure of the title compound, (displacement ellipsoids are shown at 50% probability levels). Symmetry code: (i) \u2212Click here for additional data file.10.1107/S2056989015023671/bg2577fig2.tifc . DOI: c axis, showing O\u2014H\u22efO, C\u2014H\u22efC hydrogen bonds and C\u2014H\u22ef \u03c0, and \u03c0\u2013\u03c0 stacking inter\u00adactions drawn as dotted lines.Packing view drawn along the 1014518CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, weak C\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22ef\u03c0 inter\u00adactions connect mol\u00adecules forming a three-dimensional network.In the title mol\u00adecule, C 21H20ClF2NO2, the piperidine ring adopts a slightly distorted boat conformation. The two benzene rings form a dihedral angle of 87.43\u2005(1)\u00b0. A weak intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction is observed. In the crystal, weak C\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22ef\u03c0 inter\u00adactions connect the mol\u00adecules, forming a three-dimensional network.In the title mol\u00adecule, C The N1\u2014C14 [1.356\u2005(2)\u2005\u00c5] and C14\u2014O1 [1.221\u2005(2)\u2005\u00c5] bond distances indicate the presence electron delocalization in this part of the mol\u00adecule. The six-membered piperidine ring adopts a slightly distorted boat conformation. The benzene rings form a dihedral angle of 87.43\u2005(1)\u00b0. The equatorial and axial orientation of the methyl substituents bonded to atom C2 are described by the N1\u2014C1\u2014C2\u2014C6 and N1\u2014C1\u2014C2\u2014C7 torsion angles of \u2212117.45\u2005(16)\u00b0 and \u221257.2\u2005(2)\u00b0, respectively. A weak intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction is observed, which involves the C8\u2013C13 benzene ring -t-3,t-5-di\u00admethyl\u00adpiperidin-4-one [50.4\u2005(1)\u00b0] -c-3,t-3-di\u00admethyl\u00adpiperidin-4-one [77.23\u2005(7)\u00b0] -t-3-isopropyl-1-nitro\u00adsopiperidin-4-one [21.56\u00b0] -t-3-iso\u00adpropyl\u00adpiperidin-4-one [52.4\u2005(1)\u00b0] piperidin-4-one , and tri\u00adethyl\u00adamine in benzene (20\u2005ml), di\u00adchloro\u00adacetyl\u00adchloride in benzene (20\u2005ml) was added dropwise for about half an hour. Stirring was continued with mild heating using a magnetic stirrer for 7\u2005h. The progress of the reaction was monitored by TLC. After the completion of reaction, it was poured into water and extracted with ether. The collected ether extracts were then washed well with 3% sodium bicarbonate solution and dried over anhydrous Na2SO4. The pasty mass obtained was purified by crystallization from a benzene\u2013petroleum ether solution (333\u2013353\u2005K) in the ratio of 95:5. X-ray quality crystals were grown by slow evaporation of an ethanol solution of the title compound at ambient temperature.The synthesis followed the procedure of Aridoss al. 2007. To a stUiso(H) = 1.2Ueq(C) or 1.5Ueq(Cmeth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814021278/lh5727sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814021278/lh5727Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814021278/lh5727Isup3.tifSupporting information file. DOI: Click here for additional data file.10.1107/S1600536814021278/lh5727Isup4.cmlSupporting information file. DOI: 1026047CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The ligand adopts an E conformation and the gold(I) atom displays the expected linear geometry with a Cl atom also bonded to the metal ion [Cl\u2014Au\u2014S = 174.23\u2005(5)\u00b0]. One of the pyridyl rings is protonated, giving the gold complex an overall positive charge. Two solvent water mol\u00adecules, one of which is located on a twofold rotation axis, and a non-coordinating chloride ion complete the structural assembly. The mol\u00adecular structure is stabilized by intra\u00admolecular and inter\u00admolecular N\u2014H\u22efCl, N\u2014H\u22efN, O\u2014H\u22efCl and O\u2014H\u22efO hydrogen bonding.The title complex, [AuCl(C They are compounds that can coordinate to transition metals and exhibit keto\u2013enol tautomerism of the starting material [HPy][AuClsp hybridization of the metal.The gold(I) atom displays the expected linear geometry, with a Cl\u2014Au\u2014S coordination angle of 174.23\u2005(5)\u00b0, close to the ideal angle of 180\u00b0 expected for et al., 2006The C12\u2014S1 bond length reported for di-2-pyridyl ketone phenyl\u00adthio\u00adsemicarbazone is 1.676\u2005(2)\u2005\u00c5 and it is lengthened to 1.713\u2005(4)\u2005\u00c5 on coordination to gold; this is typical of the ketone form with a concomitant shortening of the N3\u2014N4 bond in 5\u2005ml of CH3CN. A clear yellow solution was formed after heating the mixture to reflux for three\u2005h. Orange crystals deposited upon slow cooling of the solvent. Yield: 69%, m.p. 491\u2005K. Elemental analysis, found: C, 33.71; H, 3.15; N, 10.04%; calculated for C36H38Au2Cl4N10O3S2: C, 33.87; H, 3.16; N, 10.97%. IR (\u03bdmax cm\u22121): 3421 (O\u2014H), 3281 (N\u2014H), 2927 (N\u2014H+), 1694 (C=N), 1150 (N\u2014N), 765 (C=S).Di-2-pyridyl ketone phenyl\u00adthio\u00adsemicarbazone (1\u2005mmol) was dissolved in about 5\u2005ml of CHUiso = 1.5Ueq(O). Other H atoms were included in the refinement at calculated positions and treated as riding with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015015480/zl2637sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015015480/zl2637Isup3.hklStructure factors: contains datablock(s) I. DOI: 1419509CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit contains a [1-NaMePy]+ cation and one half of an Ni(imnt)22\u2212 anion. The NiII ion lies on an inversion centre and adopts a square-planar configuration with Ni\u2014S bond lengths of 2.200\u2005(1) and 2.216\u2005(1)\u2005\u00c5. In the [1-NaMePy]+ cation, the naphthyl ringsystem and the pyridinium ring make a dihedral angle of 90.0\u2005(2)\u00b0. In the crystal, C\u2014H\u22efN and C\u2014H\u22efNi hydrogen bonds, as well as \u03c0\u2013\u03c0 inter\u00adactions between the chelate ring and the pyridinium ring [centroid\u2013centroid distance = 3.675\u2005(2)\u2005\u00c5] link the ions into a three-dimensional network.A new ion-pair complex, (C Transition metal complexes with di\u00adthiol\u00adate ligands such as 2,2-di\u00adcyano\u00adethene-1,1-di\u00adthiol\u00adate (imnt) or 1,2-di\u00adcyano\u00adethene-1,2-di\u00adthiol\u00adate (mnt) are important mol\u00adecular materials with inter\u00adesting electrical conductivity, superconductivity, optical and magnetic properties 22\u2212 anion located about an inversion center. The NiS4 core exhibits a square-planar configuration, with Ni\u2014S bond lengths of 2.200\u2005(1) and 2.216\u2005(1)\u2005\u00c5. The S1\u2014Ni1\u2014S2 bond angle within the four-membered ring (Ni1/S1/C1/S2) is 78.91\u2005(3)\u00b0. The N1 and N2 atoms of the C\u00a0N groups deviate from the Ni1/S1/C1/S2 plane by 0.078\u2005(3) and 0.169\u2005(3)\u2005\u00c5, respectively. The [1-NaMePy]+ cation adopts a conformation in which both the naphthyl ring system and the pyridinium ring are twisted with respect to the N3/C11/C10 reference plane, making dihedral angles of 10.5\u2005(2)\u00b0 and 87.3\u2005(3)\u00b0, respectively. The naphthyl ring system and the pyridinium ring make a dihedral angle of 90.0\u2005(2)\u00b0.The asymmetric unit of the title compound consists of one [1-NaMePy]22\u2212 anion and [1-NaMePy]+ cation. The first is a \u03c0\u2013\u03c0 contact between the chelate ring of the anion and the pyridinium ring of the cation on Fig.\u00a02 with a don Fig.\u00a02. The comon Fig.\u00a02.22\u2212 anion have been reported, typical examples being [TBA]2[Ni(imnt)2] and [4NO2BzPy]2[Ni(imnt)2] 2] [4FBzPy is 1-(4-fluoro\u00adbenz\u00adyl)pyrid\u00adin\u00adium] 2] (Bz2NH2Py is 1-benzyl-2-amino\u00adpyridinium) 2] [BzDMAP is 1-benzyl-4-(di\u00admethyl\u00adamino)\u00adpyridinium] 2] and [2-NaMe-4-MePy]2[Ni(imnt)2] 2] and [Bz-4-MeQl]2[Ni(imnt)2] al., 2009. For a d al. 2013.2\u00b76H2O, K2imnt and 1-(4-naphthyl\u00admethyl\u00adene)pyridinium bromide in water = 0.93\u2005\u00c5, Uiso(H) = 1.2Ueq(C) for aromatic and d(C\u2014H) = 0.97\u2005\u00c5, Uiso(H) = 1.2Ueq(C) for CH2 atoms. Crystal data, data collection and structure refinement details are summarized in Table\u00a02All H-atoms were positioned geometrically and refined using a riding model with 10.1107/S1600536814017012/kp2472sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S1600536814017012/kp2472Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814017012/kp2472Isup3.cmlSupporting information file. DOI: 1015644CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In both cases, the Pd2+ cations are coordinated by the Se and N atoms of the chelating bidentate 2-[(di\u00admethyl\u00adamino)\u00admeth\u00adyl]benzene\u00adseleno\u00adlate ligand. The chloride ligand lies trans to selenium and the tri\u00adphenyl\u00adphosphane ligand is trans to nitro\u00adgen. The Pd\u2014Se bond lengths in the two independent coordination environments of Pd are 2.3801\u2005(4) and 2.3852\u2005(4)\u2005\u00c5, the Pd\u2014P bond lengths are 2.2562\u2005(8) and 2.2471\u2005(8)\u2005\u00c5, the Pd\u2014N bond lengths are 2.172\u2005(2) and 2.158\u2005(2)\u2005\u00c5, and the Pd\u2014Cl bond lengths are 2.3816\u2005(8) and 2.3801\u2005(8)\u2005\u00c5. The square-planar coordination around one Pd2+ cation is less distorted than that around the other.The asymmetric unit of the title compound, [PdCl(C DOI: 10.1107/S1600536814010678/nk2221Isup2.hklStructure factors: contains datablock(s) I. DOI: 1002116CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The backbone torsion angles for \u03b23,3-Ac6c-OH are restricted to gauche conformations in all the derivatives, with a chair conformation of the cyclo\u00adhexane ring. In the crystal structure of (I), the packing of mol\u00adecules shows both carb\u00adoxy\u00adlic acid R22(8) O\u2014H\u22efO and centrosymmetric R22(14) N\u2014H\u22efO hydrogen-bonding inter\u00adactions, giving rise to chains along the c-axis direction. In (II), centrosymmetric carb\u00adoxy\u00adlic acid R22(8) O\u2014H\u22efO dimers are extended through N\u2014H\u22efO hydrogen bonds and together with inter-ring \u03c0\u2013\u03c0 inter\u00adactions between Fmoc groups [ring centroid distance = 3.786\u2005(2)\u2005\u00c5], generate a layered structure lying parallel to (010). In the case of compound (III), carb\u00adoxy\u00adlic acid O\u2014H\u22efNpyrazine hydrogen bonds give rise to zigzag ribbon structures extending along the c-axis direction.N-Protected derivatives of 1-amino\u00adcyclo\u00adhexa\u00adneacetic acid (\u03b2 Valeroyl-\u03b23,3-Ac6c-OH (I)3,3-Ac6c-OH (II)3,3-Ac6c-OH (III)In order to investigate the effect of protecting groups and disubstitution on the conformation of \u03b2-amino acids, N-protected derivatives of 1-amino\u00adcyclo\u00adhexa\u00adneacetic acid 3,3-Ac6c-OH (II)3,3-Ac6c-OH (III)B \u2014C1A and N1\u2014C1B\u2014C1A\u2014C1\u2032) adopt a gauche conformation in all three compounds . In a 3,3-disubstituted \u03b2-amino acid residue, \u03b23,3-Ac6c-OH, the cyclo\u00adhexane ring imposes a restriction on the torsion angles \u03d5 and \u03b8. The protecting groups at the N-terminus of (I)trans geometry [\u03c90 (C4\u2014C0\u2032\u2014N1\u2014C1B) = 177.4\u2005(2) for (I)0 (O\u2014C0\u2032\u2014N1\u2014C1B) = \u2212175.64\u2005(19) for (II)0 (C6\u2014C0\u2014N1\u2014 C1A) = \u2212170.04\u2005(17)\u00b0 for (III)]. In the case of the N-protected tert-butyl\u00adoxycarbonyl (Boc) group, the protecting group adopts a cis geometry with \u03c90 = 14.50\u00b0 3,3-Ac6c-OH residue and N3 of the pyrazine ring as shown in Fig.\u00a03c. There are no intra\u00admolecular hydrogen bonding inter\u00adactions observed in the crystal structures of derivatives (I)The mol\u00adecular conformations of Valeroyl-\u03b2ii bond pairs . In (II)c). Also present in the structure are \u03c0\u2013\u03c0 inter\u00adactions between the Fmoc groups with an inter\u00adcentroid distance of 3.786\u2005(2)\u2005\u00c5. Fig.\u00a04c shows the aromatic rings of Fmoc groups stacked in a face-to-face and edge-to-face manner, together with inter-plane distances that are within the range for stabilizing \u03c0\u2013\u03c0 inter\u00adactions s Table\u00a01 give a cif Fig.\u00a04a. In 3,3Ac6c-OH was dissolved in 5\u2005ml of a 2M NaOH solution and a solution of 5\u2005mmol of valeric anhydride (931\u2005mg) dissolved in 1,4-dioxane was added, after which the mixture was stirred for 4\u2005h at room temperature. On completion of the reaction, the 1,4-dioxane was evaporated and the product was extracted with diethyl ether (3 \u00d7 5\u2005ml). The aqueous layer was acidified with 2M HCl and extracted with ethyl acetate (3 \u00d7 10ml) and the combined organic layer was washed with brine solution. The organic layer was passed over anhydrous Na2SO4 and evaporated to give Valeroyl-\u03b23Ac6c-OH . Single crystals were grown by slow evaporation from a solution in methanol/water.Preparation of Valeroyl-\u03b23,3Ac6c-OH (II)3,3Ac6c-OH was dissolved in 1M Na2CO3 solution and Fmoc-OSu dissolved in CH3CN was added. The reaction mixture was stirred at room temperature for 6\u2005h. After completion of the reaction, the CH3CN was evaporated and the residue was extracted with diethyl ether (3 \u00d7 10\u2005ml). The aqueous layer was acidified with 2M HCl and extracted with ethyl acetate (3 \u00d7 15\u2005ml). The combined organic layer was washed with brine solution. The ethyl acetate layer was passed over anhydrous Na2SO4 and evaporated. The residue was purified by crystallization in ethyl acetate/n-hexane, affording Fmoc-\u03b23,3Ac6c-OH . Single crystals were obtained by slow evaporation from an ethyl acetate/n-hexane solution.Preparation of Fmoc-\u03b23,3Ac6c-OH (III)2Cl2 and then 200\u2005\u00b5l of N-methyl\u00admorpholine was added, followed by \u03b23,3Ac6c-OMe. HCl and EDCI. HCl at 273\u2005K. The reaction mixture was stirred at room temperature for 12\u2005h. After completion of the reaction, water was added and the reaction mixture was extracted with CH2Cl2 (3 \u00d7 5ml). The combined organic layer was washed with 2M HCl (2 \u00d7 5ml), Na2CO3 (2 \u00d7 5ml) and brine solution (2 \u00d7 5ml). The organic layer was passed over anhydrous Na2SO4 and evaporated to give Pyr-\u03b23,3Ac6c-OMe . Pyr-\u03b23,3Ac6c-OMe was dissolved in 2\u2005ml of methanol and 1\u2005ml of 2M NaOH, and the reaction mixture was stirred at room temperature for 4\u2005h. Methanol was evaporated and the residue was extracted with diethyl ether (2 \u00d7 5ml). The aqueous layer was acidified with 2M HCl and extracted with ethyl acetate (3 \u00d7 5ml). The combined organic layer was washed with brine solution (1 \u00d7 5ml). The ethyl acetate layer was passed over anhydrous Na2SO4 and evaporated to give Pyr-\u03b23,3Ac6c-OH . Single crystals were grown from an ethanol/water solution.Preparation of Pyr-\u03b2Uiso values were refined. The remaining H atoms were positioned geometrically and were treated as riding on their parent C atoms, with C\u2014H distances of 0.96\u20130.98\u2005\u00c5 and with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C). For derivatives (II)Uiso values were refined. In (II)Pca21, the structure was inverted in the final cycles of refinement as the Flack parameter was 0.8\u2005(14). The inverted structure gave a value of 0.2\u2005(14) for 1585 Friedel pairs.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S1600536814020777/zs2313sup1.cifCrystal structure: contains datablock(s) I, II, III, New_Global_Publ_Block. DOI: 10.1107/S1600536814020777/zs2313Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S1600536814020777/zs2313IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S1600536814020777/zs2313IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S1600536814020777/zs2313Isup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S1600536814020777/zs2313IIsup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S1600536814020777/zs2313IIIsup7.cmlSupporting information file. DOI: 1024488, 1024489, 1024490CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "By contrast to this almost 17H14PS)2], is a second monoclinic polymorph of the previously reported monoclinic form [Fang et al. \u00b0. By contrast to this almost syn disposition, in the C2/c polymorph, the Fe atom lies on a centre of inversion so that the S atoms are strictly anti, with a pseudo-S\u2014P\u22efP\u2014S torsion angle of 180\u00b0. The significant difference in mol\u00adecular conformation between the two forms does not result in major perturbations in the P=S bond lengths nor in the distorted tetra\u00adhedral geometries about the P atoms. The crystal packing of the new monoclinic polymorph features weak Cp\u2014C\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adactions consolidating linear supra\u00admolecular chains along the a axis. These pack with no directional inter\u00adactions between them.The title compound, [Fe(C al. 1995. Polyhed R3PAu(S2CNR\u20322), attract on-going inter\u00adest owing to impressive biological activities against both cancer ferrocene (dppf), were isolated as orange needles, being a side-product of a reaction, see Synthesis and crystallization for details. Crystallography shows the title compound to be a new monoclinic polymorph of a previously described C2/c form di\u00adthio\u00adcarbamates, 2 is shown in Fig.\u00a012P=S units linked via the P atoms through a C5H4FeC5H4 link. The S atoms lie to the same side of the mol\u00adecule and might be described as having a syn conformation. When viewed down the P\u22efP axis, the S atoms are gauche with the pseudo S\u2014P\u22efP\u2014S torsion angle being \u221253.09\u2005(3)\u00b0. This represents the major difference between dppfS2 and its C2/c\u2013dppfS2 polymorph and Cg(C6\u2013C10) are 1.6487\u2005(8) and 1.6451\u2005(8)\u2005\u00c5, respectively, and the Cg(C1\u2013C5)\u22efFe\u22efCg(C6\u2013C10) angle is 178.92\u2005(5)\u00b0. The comparable parameters for the C2/c\u2013dppfS2 polymorph are 1.650\u2005(3)\u2005\u00c5 and 180\u00b0, and the Cp rings are strictly staggered when viewed down the Cg(C1\u2013C5)\u22efFe\u22efCg(C1\u2013C5)i axis. In dppfS2, the P=S bond lengths are experimentally distinct, i.e. P1=S1 of 1.9449\u2005(6)\u2005\u00c5 is shorter than P2=S2 of 1.9530\u2005(6)\u2005\u00c5, with the former being equivalent to P1=S1 of 1.9384\u2005(18)\u2005\u00c5 in C2/c\u2013dppfS2. Finally, the P1 and P2 atoms have distorted tetra\u00adhedral environments with the range of angles subtended at P1 of 103.94\u2005(7)\u2013113.78\u2005(6)\u00b0 being comparable to those subtended at P2, i.e. 105.55\u2005(7)\u2013114.92\u2005(5)\u00b0; the equivalent range of angles in C2/c\u2013dppfS2 is 104.8\u2005(2)\u2013114.28\u2005(15)\u00b0. In each case, the angles involving the S atom are wider than those involving C atoms only, and the narrowest angle always involves the two ipso-C atoms.The conformational differences in the polymorphs are highlighted in the overlay diagram shown in Fig.\u00a02a axis. Based on the distance criteria employed in PLATON C5H4FeC5H4P(=Y)Ph2, Y = 0, O, S and Se, have been described in the crystallographic literature. The parent compound, i.e. with Y = lone pair, has the Fe atom situated on a centre of inversion C5H4FeC5H4P(=Y)Ph2, Y = 0, O, S and Se, compounds suggesting a low energy barrier for the inter\u00adchange from one conformation to another. The structural data for Ph2P(=Y)C5H4FeC5H4P(=Y)Ph2 are summarized in Table\u00a02The structures of several oxidation products of dppf, Ph2 mol\u00adecule can function as a ligand in metal complexes, often forming zero-dimensional mononuclear species and CS2 (84.6\u2005\u00b5l) were added. Chloro\u00adform (150\u2005ml) was then added and the reaction mixture was stirred for 2\u2005h. A second solution containing bis\u00ad[chlorido\u00adgold(I)] (1.4\u2005mmol) was prepared by dissolving potassium tetra\u00adchlorido\u00adaurate(III) (1.06\u2005g) in a solvent mixture of acetone and water . Drop-wise addition of sodium sulfite (0.71\u2005g) in water (10\u2005ml) followed. Upon discolouration, bis\u00ad(di\u00adphenyl\u00adphos\u00adphane)ferrocene in chloro\u00adform (25\u2005ml) was added. After stirring for 15 mins, the resulting gold precursor was extracted with chloro\u00adform (150\u2005ml). Aceto\u00adnitrile (50\u2005ml) was added to this to form solvent mixture of chloro\u00adform and aceto\u00adnitrile (3:1). The solution containing the di\u00adthio\u00adcarbamate was added to that containing the gold precursor. The resulting mixture was stirred for 3\u2005h. and then filtered. After three weeks, orange needles appeared, along with the precipitate, and these were subjected to the crystallographic study. Yield: 0.0890\u2005g, 10.3% (based on dppf). M.p.: 519.5\u2013519.9\u2005K. IR: \u03bd(P=S) 628 (m).Two solutions were prepared. Firstly, a solution sodium salt of piperazine di\u00adthio\u00adcarbamate (0.7\u2005mmol) was prepared by dissolving piperazine (0.0582\u2005g) in aceto\u00adnitrile (50\u2005ml). NaOH (112\u2005\u00b5l of 50% Uiso(H) set to 1.2Uequiv(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015012682/hg5450sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015012682/hg5450Isup2.hklStructure factors: contains datablock(s) I. DOI: 1409866CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the canagliflozin mol\u00adecules and lattice water mol\u00adecules are connected A and B) and one water mol\u00adecule in the asymmetric unit of the title compound, C24H25FO5S\u00b70.5H2O [systematic name: -2-(3-{[5-(4-fluoro\u00adphen\u00adyl)thio\u00adphen-2-yl]meth\u00adyl}-4-methylphen\u00adyl)-6-(hy\u00addroxy\u00admeth\u00adyl)-3,4,5,6-tetra\u00adhydro-2H-pyran-3,4,5-triol hemihydrate]. The dihedral angles between the methyl\u00adbenzene and thio\u00adphene rings are 115.7\u2005(4) and 111.7\u2005(4)\u00b0, while the dihedral angles between the fluoro\u00adbenzene and thio\u00adphene rings are 24.2\u2005(6) and 20.5\u2005(9)\u00b0 in mol\u00adecules A and B, respectively. The hydro\u00adpyran ring exhibits a chair conformation in both canagliflozin mol\u00adecules. In the crystal, the canagliflozin mol\u00adecules and lattice water mol\u00adecules are connected via O\u2014H\u22efO hydrogen bonds into a three-dimensional supra\u00admolecular architecture.There are two canagliflozin mol\u00adecules ( The conformational difference is also shown by the angle C10\u2014C11\u2014C12, which is 115.7\u2005(4)\u00b0 in mol\u00adecule A and 111.7\u2005(4)\u00b0 in mol\u00adecule B. The terminal aromatic rings (C1\u2013C6) are inclined to the thio\u00adphene rings, forming dihedral angles of 24.2\u2005(6) and 20.5\u2005(9)\u00b0 in mol\u00adecules A and B, respectively. The tetra\u00adhydro\u00adpyran rings exhibit a distorted chair conformation in both mol\u00adecules A and B.The conformations of the two canagliflozin mol\u00adecules are somewhat different with regard to the orientation of the central benzene ring (C12\u2013C17) with respect to the thio\u00adphene ring, as indicated by torsion angles C9B\u2014H3B1\u22efO4Bi, O2B\u2013H2B1\u22efO4Aiii, and O5B\u2014H5B1\u22efO3Biv link canagliflozin mol\u00adecules, generating a ring of graph-set motif B\u2014H4B\u22efO6, O6\u2014H61\u22efO2A and O4A\u2014H4A\u22efO5Bii hydrogen bonds propagating along the a axis; the chains are stacked along the c axis by further hydrogen-bonding inter\u00adactions, O3A\u2014H3A1\u22efO2Bi and O2A\u2013-H2A1\u22efO2Bi = 1.2Ueq or 1.5Ueq(carrier atom).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016006769/xu5886sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016006769/xu5886Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016006769/xu5886Isup3.cmlSupporting information file. DOI: 1475516CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title complex is composed of saddle-shaped mol\u00adecules which closely inter\u00adact in a pairwise fashion through \u03c0\u2013\u03c0 and C\u2014H\u22ef\u03c0 contacts to form \u2018dimers\u2019. These \u2018dimers\u2019 further inter\u00adact through C\u2014H\u22efS and C\u2014H\u22ef\u03c0 contacts to construct a complex three-dimensional extended structure. 20H12S4, (I), also known as exTTF, is reported. The mol\u00adecular structure of (I) consists of a di\u00adhydro\u00adanthracene moiety with two 1,3-di\u00adthiol-2-yl\u00adidene substituents. This is a saddle-shaped mol\u00adecule, which inter\u00adacts with a close neighbor through various \u03c0\u2013\u03c0 and C\u2014H\u22ef\u03c0 contacts to form a \u2018dimer\u2019. These \u2018dimers\u2019 inter\u00adact through a series of C\u2014H\u22efS and C\u2014H\u22ef\u03c0 contacts to construct a complex three-dimensional extended structure.The crystal structure of the well-studied 9,10-bis\u00ad-9,10-di\u00adhydro\u00adanthracene mol\u00adecule, C The mol\u00adecule is saddle shaped in that the 1,3-di\u00adthiol-2-yl\u00adidene groups bend significantly up out of the plane of the central ring and the two benzene rings of di\u00adhydro\u00adanthracene moiety bend down out of the plane, Fig.\u00a01b. The central six-membered ring (C4\u2013C5\u2013C10\u2013C11\u2013C12\u2013C17) is in a boat conformation in which the 1,3-di\u00adthiol-2-yl\u00adidene-substituted carbon atoms [C4 and C11] are bent out of the plane defined by C5, C10, C12, and C17. The torsion angles C10\u2014C12\u2014C17\u2014C4 = 17.97\u2005(12)\u00b0 and C17\u2014C5\u2014C10\u2014C11 = 17.22\u2005(16)\u00b0 for these two carbon atoms are quite similar.The mol\u00adecular structure of (I)The benzene rings bend out of the C5\u2013C10\u2013C12\u2013C17 plane; the dihedral angle between this plane and the plane of the C5\u2013C6\u2013C7\u2013C8\u2013C9\u2013C10 ring is 17.72\u2005(15)\u00b0 while the dihedral angle for the C12\u2013C13\u2013C14\u2013C15\u2013C16\u2013C17 ring is 20.14\u2005(13)\u00b0. The 1,3-di\u00adthiol-2-yl\u00adidene groups are bent more sharply out of the C5-C10-C12-C17 plane as evidenced by the torsion angles C3\u2014C4\u2014C5\u2014C10 \u03c4 = 138.06\u2005(15)\u00b0 and C18\u2014C11\u2014C12\u2014C17 \u03c4 = 139.23\u2005(15)\u00b0. The five-membered rings both adopt an envelope conformation with the carbon atom bonded to the di\u00adhydro\u00adanthracene [C3 and C18] being the one puckered out of the plane. The torsion angles C3\u2014S1\u2014C1\u2014C2 \u03c4 = \u22128.09\u2005(14)\u00b0 and C18\u2014S4\u2014C20\u2014C19 \u03c4 = \u22126.65\u2005(15)\u00b0 show that the bend in each ring is fairly similar.The average C\u2014C bond length within the benzene rings is 1.391\u2005\u00c5 as is typical of phenyl rings. The length of the edges shared with the central ring are slightly longer C5\u2014C10 = 1.419\u2005(2)\u2005\u00c5 and C12\u2014C17 = 1.412\u2005(2)\u2005\u00c5. The remaining C\u2014C distances making up the central ring are longer still with an average of 1.477\u2005\u00c5. Since the distances within the central ring are in between those of typical C\u2014C single and double bonds; this supports the idea of a highly delocalized bonding motif throughout the di\u00adhydro\u00adanthracene ring system. The bond distances between the di\u00adhydro\u00adanthracene and the 1,3-di\u00adthiol-2-yl\u00adidene groups are on the order of typical C=C bonds, C3=C4 = 1.360\u2005(2)\u2005\u00c5 and C11=C18 = 1.361\u2005(2)\u2005\u00c5.i\u2013C2i\u2013S2i\u2013C3i\u2013S1i ring and is rather long at 4.068\u2005(15)\u2005\u00c5. There are five C\u2014H\u22ef\u03c0 inter\u00adactions between the two mol\u00adecules in which atoms H1 and H2 of one mol\u00adecule inter\u00adact with various \u03c0 systems of the neighbor. The shortest contact is between H1 and the C11i=C18i double bond at 2.606\u2005(12)\u2005\u00c5 . There is another short contact between H1 and the central ring of the di\u00adhydro\u00adanthracene, H1\u22efcentroid (C4i\u2013C5i\u2013C10i\u2013C11i\u2013C12i\u2013C17i) 2.852\u2005(11)\u2005\u00c5. Two other C\u2014H\u22ef\u03c0 inter\u00adactions involve H1; H1\u22efcentroid (C18i\u2013S3i\u2013C19i\u2013C20i\u2013S4i) 3.167\u2005(11)\u2005\u00c5, and H1\u22efcentroid (C5i\u2013C6i\u2013C7i\u2013C8i\u2013C9i\u2013C10i) 3.553\u2005(15)\u2005\u00c5. The fifth inter\u00adaction between the \u2018dimer\u2019 mol\u00adecules is H2\u22efcentroid (C5i\u2013C6i\u2013C7i\u2013C8i\u2013C9i\u2013C10i) 3.222\u2005(12)\u2005\u00c5.Through a series of C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions, each mol\u00adecule of (I)ii 2.922\u2005(12)\u2005\u00c5 and one C\u2014H\u22ef\u03c0 contact H14\u22efcentroid (C5ii\u2013C6ii\u2013C7ii\u2013C8ii\u2013C9ii\u2013C10ii) 3.779\u2005(17)\u2005\u00c5 . H15 also inter\u00adacts with two groups on a neighboring mol\u00adecule through two C\u2014H\u22ef\u03c0 contacts; H15\u22efcentroid (C12iii\u2013C13iii\u2013C14iii\u2013C15iii\u2013C16iii\u2013C17iii) 3.385\u2005(17)\u2005\u00c5 and H15\u22efcentroid (C4iii\u2013C5iii\u2013C10iii\u2013C11iii\u2013C12iii\u2013C17iii) 3.543\u2005(14)\u2005\u00c5 . It should be noted that the mol\u00adecules generated by symmetry operations (ii) and (iii) form a \u2018dimer\u2019. The final inter\u00adaction involving a CH group on the di\u00adhydro\u00adanthracene is H6\u22efcentroid (S1iv\u2013C1iv\u2013C2iv\u2013S2iv\u2013C3iv) 2.865\u2005(11)\u2005\u00c5 . Taking these inter\u00adactions into account, a two-dimensional layered structure is formed ed Fig.\u00a03 in whichv\u2013C6v\u2013C7v\u2013C8v\u2013C9v\u2013C10v) 2.829\u2005(18)\u2005\u00c5, H19\u22efcentroid (C4v\u2013C5v\u2013C10v\u2013C11v\u2013C12v\u2013C17v) 3.301\u2005(11)\u2005\u00c5, and H20\u22efcentroid (C12v\u2013C13v\u2013C14v\u2013C15v\u2013C16v\u2013C17v) 2.767\u2005(11)\u2005\u00c5 . These hydrogen atoms also inter\u00adact with another mol\u00adecule via C\u2014H\u22efS contacts; H19\u22efS4vi 3.367\u2005(12)\u2005\u00c5 and H20\u22efS3vi 3.288\u2005(14)\u2005\u00c5 [symmetry operation: (vi) \u2212x, y\u00a0+\u00a00.5, \u2212z\u00a0+\u00a0a axis to form a three-dimensional extended structure, Fig.\u00a04There are also five C\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions in which the CH group involved resides on the 1,3-di\u00adthiol-2-yl\u00adidene portion of (I)et al., 1990P21/c) and has a similar saddle shape. It also appears to form similar \u2018dimers\u2019 in which there are both C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions between the two mol\u00adecules.Many derivatives of (I)\u22121 less stable than the predicted favorite. The study details \u03c0\u2013\u03c0 stacking between two of the di\u00adthiol rings, C\u2014H\u22ef\u03c0 contacts between the di\u00adthiol H atoms and the anthracene rings, \u03c0\u2013\u03c0 stacking between anthracene units, as well as an inter\u00adaction between the partial positive charge of the S atoms and the anthracene rings for the preferred computational \u2018dimer\u2019. The study briefly describes the C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions found in (I)A recent computational study focused on predicting the most energetically favored \u2018dimers\u2019 of (I)et al., 1989The title complex, 9,10-bis\u00ad-9,10-di\u00adhydro\u00adanthracene (I)Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2056989015020800/bg2573sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015020800/bg2573Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015020800/bg2573Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015020800/bg2573Isup4.cmlSupporting information file. DOI: 1434765CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The components of the 2:1 co-crystal are linked by hy\u00addroxy-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonds into a three-mol\u00adecule aggregate having the shape of the letter Z. These are connected into a supra\u00admolecular ladder by tight amide-N\u2014H\u22efO(nitro) hydrogen bonds. 7H5NO4\u00b7C14H14N4O2, in which the complete di\u00adamide mol\u00adecule is generated by crystallographic inversion symmetry, features a three-mol\u00adecule aggregate sustained by hydroxyl-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonds. The p-nitro\u00adbenzoic acid mol\u00adecule is non-planar, exhibiting twists of both the carb\u00adoxy\u00adlic acid and nitro groups, which form dihedral angles of 10.16\u2005(9) and 4.24\u2005(4)\u00b0, respectively, with the benzene ring. The di\u00adamide mol\u00adecule has a conformation approximating to a Z shape, with the pyridyl rings lying to either side of the central, almost planar di\u00adamide residue (r.m.s. deviation of the eight atoms being 0.025\u2005\u00c5), and forming dihedral angles of 77.22\u2005(6)\u00b0 with it. In the crystal, three-mol\u00adecule aggregates are linked into a linear supra\u00admolecular ladder sustained by amide-N\u2014H\u22efO(nitro) hydrogen bonds and orientated along [10-4]. The ladders are connected into a double layer via pyridyl- and benzene-C\u2014H\u22efO(amide) inter\u00adactions, which, in turn, are connected into a three-dimensional architecture via \u03c0\u2013\u03c0 stacking inter\u00adactions between pyridyl and benzene rings [inter-centroid distance = 3.6947\u2005(8)\u2005\u00c5]. An evaluation of the Hirshfeld surfaces confirm the importance of inter\u00admolecular inter\u00adactions involving oxygen atoms as well as the \u03c0\u2013\u03c0 inter\u00adactions.The title 2:1 co-crystal, 2C The results of this investigation are reported herein.Arguably, the most prominent motivation for the study of co-crystals relates to their potential applications in the pharmaceutical industry whereby co-crystals of active pharmaceutical ingredients (APIs) formed with generally regarded as safe (GRAS) co-crystal coformers might provide drugs with enhanced useful properties, p-nitro\u00adbenzoic acid mol\u00adecule in a general position, and a N,N\u2032-bis\u00ad(pyridin-3-ylmeth\u00adyl)ethanedi\u00adamide mol\u00adecule situated about a centre of inversion. This results in the 2:1 co-crystal stoichiometry.The title co-crystal, Fig.\u00a01A2/a: Tonogaki et al., 1993P21/m: Bolte, 2009Twists are noted in the acid mol\u00adecule so that the dihedral angle between the benzene ring and the non-hydrogen atoms of the carb\u00adoxy\u00adlic acid group is 10.16\u2005(9)\u00b0. The comparable angle involving the nitro group is 4.24\u2005(4)\u00b0, consistent with a smaller twist. The substituents have a conrotatory disposition forming a dihedral angle of 13.50\u2005(8)\u00b0. The crystal structure of the free acid was first reported almost fifty years ago \u2005\u00c5 in the title co-crystal.The di\u00adamide features an essentially flat central residue with the r.m.s. deviation for the eight non-hydrogen atoms being 0.025\u2005\u00c5. This planar arrangement allows for the formation of intra\u00admolecular amide-N\u2014H\u22efO(amide) hydrogen bonds Table\u00a02. The pyret al., 20085NO\u22efHNC2NC3N}2 supra\u00admolecular rings, Fig.\u00a02i.e. Cg(pyrid\u00adyl)\u22efCg(benzene)i = 3.7214\u2005(8)\u2005\u00c5, with an angle of 4.69\u2005(7)\u00b0 between the rings; symmetry operation: (i) 2\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z. The connections between the layers to consolidate the three-dimensional architecture, Fig.\u00a04i.e. Cg(pyrid\u00adyl)\u22efCg(benzene)ii = 3.6947\u2005(8)\u2005\u00c5 with an angle of inclination = 4.69\u2005(7)\u00b0; symmetry operation: (ii) 2\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z.In the packing, the anti\u00adcipated inter\u00adaction and the similar long spike at the same ed + id distance in the lower right region indicates the contribution of the amide-N\u2014H\u22efO(nitro) inter\u00adaction. The donor\u2013acceptor contributions of these co-crystal constituents are highlighted with the label \u2018d\u2032 in the fingerprint plot, Fig.\u00a07i.e. 32.3%, to the Hirshfeld surface of the acid, a smaller contribution, i.e. 8.3%, is provided by the di\u00adamide; the reverse is true for for the O\u22efH contacts, i.e. 8.5 and 30.4%, respectively. The overall fingerprint plot for the co-crystal when delineated into O\u22efH/H\u22efO contacts leads to the pair of spikes corresponding to donors and acceptors with a 37.1% contribution to surface and amide-N\u2014H\u22efO(nitro) inter\u00adactions between the pair of acid mol\u00adecules and a di\u00adamide mol\u00adecule can be observed through their corresponding Hirshfeld surfaces mapped over the electrostatic potential, Fig.\u00a05i.e. the enrichment ratio, ER hydrogen bonding was observed hydrogen bonding is seen along with hy\u00addroxy-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonding 2C6H3CO2\u2212 anion , prepared in accord with the literature procedure (Schauer Uiso(H) set to 1.2Uequiv(C). The oxygen- and nitro\u00adgen-bound H-atoms were located in a difference Fourier map but were refined with distance restraints of O\u2014H = 0.84\u00b10.01\u2005\u00c5 and N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.5Ueq(O) and 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989015024068/hb7557sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015024068/hb7557Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015024068/hb7557Isup3.cmlSupporting information file. DOI: 1442547CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title chalcone derivative, mol\u00adecules are linked into a three-dimensional network by C\u2014H\u22efO hydrogen bonds and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions are also observed. The inter\u00admolecular inter\u00adactions in the crystal structure were qu\u00adanti\u00adfied and analysed using Hirshfeld surface analysis. 16H11ClF2O2, the enone group adopts an E conformation. The dihedral angle between the benzene rings is 0.47\u2005(9)\u00b0 and an intra\u00admolecular C\u2014H\u22efF hydrogen bond closes an S(6) ring. In the crystal, mol\u00adecules are linked into a three-dimensional network by C\u2014H\u22efO hydrogen bonds and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions are also observed [centroid\u2013centroid separation = 3.5629\u2005(18)\u2005\u00c5]. The inter\u00admolecular inter\u00adactions in the crystal structure were qu\u00adanti\u00adfied and analysed using Hirshfeld surface analysis.In the title chalcone derivative, C The mol\u00adecule is slightly twisted at the C9\u2014-C10 bond with a C8\u2014C9\u2014C10\u2014C15 torsion angle of \u22122.2\u2005(4)\u00b0 and a maximum deviation of 0.193\u2005(16)\u2005\u00c5 for atom O1. The dihedral angle between the terminal benzene rings (C1\u2013C6 and C10\u2013C15) is 0.47\u2005(9)\u00b0. The least-squares plane through the enone moiety (O1/C7\u2013C9) makes dihedral angles of 2.87\u2005(14) and 3.33\u2005(14)\u00b0 with the C1\u2013C6 and C10\u2013C15 benzene rings, respectively. An intra\u00admolecular C8\u2014H8A\u22efF1 hydrogen bond (Table\u00a01S(6) ring motif. The bond lengths and angles are comparable with the equivalent data for previously reported structures; d Table\u00a01 is observia C2\u2014H2A\u22efO1 , centroid-to-centroid distance = 3.5629\u2005(18)\u2005\u00c5, where Cg1 and Cg2 are the centroids of the C1\u2013C6 and C10\u2013C15 rings, respectively].In the crystal, mol\u00adecules are linked into a three-dimensional network s Table\u00a01, as showCrystal Explorer 3.1 with neighbouring mol\u00adecules connected by C2\u2014H2A\u22efO1 and C3\u2014H3A\u22efO2 hydrogen bonds. This finding is corroborated by Hirshfeld surfaces mapped over the electrostatic potential showing the negative potential around the oxygen atoms as light-red clouds and the positive potential around hydrogen atoms as light-blue clouds.The strong C\u2014H\u22efO inter\u00adactions are visualized as bright-red spots between the respective donor and acceptor atoms on the Hirshfeld surfaces mapped over rm Fig.\u00a03a with nal Fig.\u00a03b showined = id \u223c1.4\u2005\u00c5 with overall Hirshfeld surfaces of 27.5%. The contribution from the O\u22efH/H\u22efO contacts, corresponding to C\u2014H\u22efO inter\u00adactions, is represented by a pair of sharp spikes characteristic of a strong hydrogen-bond inter\u00adaction having almost the same ed + id \u223c2.3\u2005\u00c5 .Significant inter\u00admolecular inter\u00adactions are plotted in Fig.\u00a04\u2005\u00c5 Fig.\u00a04a with o\u2005\u00c5 Fig.\u00a04b.ed = id \u223c1.8\u2005\u00c5 . The presence of the \u03c0\u2013\u03c0 stacking inter\u00adactions is also indicated by the appearance of red and blue triangles on the shape-indexed surfaces, identified with black arrows in Fig.\u00a05The C\u22efC contacts, which refer to \u03c0\u2013\u00b7\u03c0 stacking inter\u00adactions, contribute 13.7% of the Hirshfeld surfaces. This appears as a distinct triangle at around \u2005\u00c5 Fig.\u00a04c. The pA mixture of 3-fluoro-4-meth\u00adoxy\u00adaceto\u00adphenone and 2-chloro-6-fluoro\u00adbenzaldehyde was dissolved in methanol (20\u2005ml). A catalytic amount of NaOH was added to the solution dropwise with vigorous stirring. The reaction mixture was stirred for about 5\u20136\u2005h at room temperature. After stirring, the contents of the flask were poured into ice-cold water (50\u2005ml) and the resulting crude solid was collected by filtration. Brownish blocks of (I)Uiso(H) = 1.2Ueq(C). In the final refinement, the most disagreeable reflection (020) was omitted.Crystal data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016006526/hb7578sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016006526/hb7578Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016006526/hb7578Isup3.cmlSupporting information file. DOI: 1474605CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In [Cu(Hspar)2](bdc)\u00b72H2O,the Cu2+ ion lies on a crystallographic inversion centre and a CuO4 square-planar geometry arises from its coordination by two O,O\u2032-bidentate Hspar mol\u00adecules. The bdc dianion acts as a counter-ion to the cationic complex and does not bond to the metal ion.The Mn 2O,O\u2032)bis\u00admanganese(II) dihydrate, [Mn(C8H4O4)(C19H22F2N4O3)2(H2O)0.25]\u00b72H2O or [Mn(bdc)(Hspar)2(H2O)0.25]\u00b72H2O, (I), and bis\u00adcopper(II) benzene-1,4-di\u00adcarboxyl\u00adate dihydrate, [Cu(C19H22F2N4O3)2](C8H4O4)\u00b72H2O or [Cu(Hspar)2](bdc)\u00b72H2O, (II), are reported . The Mn2+ ion in (I) is coordinated by two O,O\u2032-bidentate Hspar neutral mol\u00adecules (which exist as zwitterions) and an O,O\u2032-bidentate bdc dianion to generate a distorted MnO6 trigonal prism. A very long bond [2.580\u2005(12)\u2005\u00c5] from the Mn2+ ion to a 0.25-occupied water mol\u00adecule projects through a square face of the prism. In (II), the Cu2+ ion lies on a crystallographic inversion centre and a CuO4 square-planar geometry arises from its coordination by two O,O\u2032-bidentate Hspar mol\u00adecules. The bdc dianion acts as a counter-ion to the cationic complex and does not bond to the metal ion. The Hspar ligands in both (I) and (II) feature intra\u00admolecular N\u2014H\u22efO hydrogen bonds, which close S(6) rings. In the crystals of both (I) and (II), the components are linked by N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, generating three-dimensional networks.The syntheses and crystal structures of 0.25-aqua\u00ad(benzene-1,4-di\u00adcarboxyl\u00adato-\u03ba As well as their biological significance, this class of compounds is of inter\u00adest in coordination chemistry due to their potential to act as multi-dentate and bridging ligands in the construction of mononuclear and dinuclear complexes complexes with nickel with BF4\u2212 0.25]\u00b72H2O (I)2](bdc)\u00b72H2O (II)8H4O42\u2212).As a continuation of these studies, we now describe the syntheses and crystal structures of the title mixed-ligand complexes Mn\u2014O bond to the partly occupied O13 water mol\u00adecule. Together, these lead to a distorted MnO6+1 trigonal\u2013prismatic polyhedron n Table\u00a01 with theds Fig.\u00a02. The meads Fig.\u00a02 for the 2 groups mutually syn.The conformation of the \u2013O2\u2013C1\u2013C2\u2013C3\u2013O3\u2013Mn1\u2013 chelate ring approximates to a shallow envelope with the metal atom as the flap, displaced by \u22120.222\u2005(4)\u2005\u00c5 from the mean plane of the ligand atoms (r.m.s. deviation = 0.022\u2005\u00c5). The \u2013O5\u2013C20\u2013C21\u2013C22\u2013O6\u2013Mn1\u2013 ring can be described in the same way, with Mn1 displaced by 0.128\u2005(4)\u2005\u00c5 from the other atoms (r.m.s. deviation = 0.019\u2005\u00c5). The dihedral angle between the near-planar segments of the chelate rings is 29.74\u2005(13)\u00b0. Both Hspar mol\u00adecules are orientated in the same sense with respect to the metal ion, with the NH6+1 grouping is unusual and calls for some further comment: the dihedral angle between the top (O3/O6/O8) and bottom (O2/O5/O7) triangular faces of the prism is 14.40\u2005(11)\u00b0, which is largely due to the O7\u22efO8 edge of the prism (the two O atoms of the bdc dianion) being much shorter [2.174\u2005(3)\u2005\u00c5] than the O2\u22efO3 [2.799\u2005(3)\u2005\u00c5] and O5\u22efO6 [2.802\u2005(3)\u2005\u00c5] edges, which correspond to the C1- and C-20 Hspar mol\u00adecules, respectively. The metal atom is displaced from the top and bottom faces of the prism by \u22121.2513\u2005(14) and 1.3670\u2005(12)\u2005\u00c5, respectively. The degree of twist of the prism may be estimated from the pseudo torsion angles involving the centroids of the triangular faces (denoted X1 for the O3/O6/O8 face and X2 for the O2/O5/O7 face) and the pairs of atoms forming the edges of the prism: values of X1\u22efO7\u22efO8\u22efX2 (\u201314.6), X1\u22efO5\u22efO6\u22efX2 (\u201311.2) and X1\u22efO2\u22efO3\u22efX2 (\u20138.5\u00b0) arise. These angles would be zero for a perfect triangular prism.The capped trigonal\u2013prismatic geometry of the MnOsp2 hybridization for this atom. The dihedral angle between the N2 ring and the C5 ring (r.m.s. deviation = 0.028\u2005\u00c5), which are fused at the C4\u2014C9 bond, is 7.9\u2005(2)\u00b0, indicating a substantial puckering to the quinolone system. The piperazinium ring adopts a typical chair conformation with the exocyclic N\u2014Cq (q = quinolone) bond in an equatorial orientation. The dihedral angle between the four C atoms that form the \u2018seat\u2019 of the chair and the C5 ring is 60.3\u2005(2)\u00b0. There was some suggestion that atoms C14 and C17 of this ring are positionally disordered, but refinements that attempted to model this effect were inconclusive.The most important geometrical features of the first Hspar mol\u00adecule (containing C1) are as follows: the C1\u2014O1 and C1\u2014O2 bond lengths of 1.251\u2005(4) and 1.256\u2005(4)\u2005\u00c5, respectively, are typical for a delocalized carboxyl\u00adate group and the dihedral angle between C1/O1/O2 and the adjacent N2-containing ring (r.m.s. deviation = 0.045\u2005\u00c5) is 8.6\u2005(8)\u00b0. The dihedral angle between the cyclo\u00adpropane ring and the N2 ring is 67.5\u2005(3)\u00b0. The N2 bond-angle sum of 359.8\u00b0 is consistent with a bonding model of S(6) ring. The C45/O7/O8 and C46/O9/O10 carboxyl\u00adate groups of the bdc dianion are rotated by 3.90\u2005(7) and 25.28\u2005(14)\u00b0, respectively with respect to the central ring plane. The O7\u2014Mn1\u2014O8 bite angle is 56.58\u2005(8)\u00b0.The second Hspar mol\u00adecule (containing C20) has a broadly similar geometry: the C20\u2014O4 and C20\u2014O5 bond lengths are 1.254\u2005(4) and 1.257\u2005(4)\u2005\u00c5, respectively, and the dihedral angle between C20/O4/O5 and the N6 ring (r.m.s. deviation = 0.050\u2005\u00c5) is 8.8\u2005(7)\u00b0. The dihedral angle between the N6 (bond-angle sum = 359.7\u00b0) ring and the pendent three-membered ring is 69.8\u2005(2)\u00b0. The N6 and C24 rings (r.m.s. deviation for the latter = 0.020\u2005\u00c5), fused at the C23\u2014C28 bond, are tilted by 8.1\u2005(2)\u00b0. The piperazine ring adopts a chair conformation and the dihedral angle between the chair seat and the C24 ring is 58.71\u2005(9)\u00b0. Each Hspar mol\u00adecule features an intra\u00admolecular N\u2014H\u22efO hydrogen bond Table\u00a02, which c2+ cation lying on a crystallographic inversion centre, a neutral, zwitterionic, Hspar mol\u00adecule, half a bdc dianion and a water mol\u00adecule of crystallization on Fig.\u00a03.O,O-bidentate Hspar mol\u00adecules in the usual bonding mode of quinoline O atom + syn-carboxyl\u00adate O atom with a bite angle of 93.24\u2005(8)\u00b0, which generates a six-membered chelate ring. The result is a CuO4 square-planar coordination polyhedron (Table\u00a03The copper ion in (II)n Table\u00a03 with a mIn the Hspar mol\u00adecule, the C1\u2014O1 and C1\u2014O2 bond lengths are distinctly different at 1.226\u2005(4)\u2005\u00c5 and 1.283\u2005(4)\u2005\u00c5, respectively, unlike the situation in (I)In the bdc dianion, the C23/O4/O5 carboxyl\u00adate group is rotated by 2.7\u2005(6)\u00b0 with respect to the aromatic ring plane. The C23\u2014O4 and C23\u2014O5 bond lengths of 1.244\u2005(4) and 1.253\u2005(4)\u2005\u00c5, respectively, are consistent with the approximately equal delocalization of the negative charge over both C\u2014O bonds.In the crystal of (I)In (II)2+ ions and Hspar mol\u00adecules. The O,O-chelating mode of the Hspar mol\u00adecules is normal for other divalent transition metals ] : as these authors note, the high-spin d5 electronic configuration of Mn2+ is the \u2018least unexpected\u2019 to show a trigonal\u2013prismatic geometry because it has no crystal-field stabilization energy, which normally favours octa\u00adhedral over trigonal\u2013prismatic geometry ] have almost the same energy and the trigonal\u2013prismatic geometry is adopted in the crystal because of favourable packing inter\u00adactions 2]2+ cations seen here. In [Cu(H2spar)(H2O)(phen)]BF4\u00b73H2O and the N,N-bidentate phen ligand in a square-planar arrangement; the water mol\u00adecule completes the square-based pyramidal coordination polyhedron in the apical site. Finally, in the novel bimetallic complex [Cu2(spar)4]\u00b74H2O \u2005\u00c5] arising from the \u2013NH2 group of an adjacent spar\u2212 anion generating a centrosymmetric, bimetallic assembly. It is thus notable that sparfloxacin can bind to Cu2+ ions in its anionic, neutral and cationic forms and we are continuing our explorations of these systems.Compound (II)3CO2)2\u00b74H2O (0.25\u2005mmol), sparfloxacin (0.5\u2005mmol), 1,4-benzene\u00addicarb\u00adoxy\u00adlic acid (0.25\u2005mmol), sodium hydroxide (1\u2005mmol) and water (15\u2005ml) was stirred for 30 minutes in air. The mixture was placed in a sealed 25\u2005ml Teflon-lined hydro\u00adthermal reactor and heated to 423\u2005K for 72\u2005h under autogenous pressure. Upon cooling, colourless prisms of (I)46H52.5MnF4N8O12.25: C 52.90 (52.63), H 5.07 (4.91), N 10.73 (10.58). IR : br3420, br3300, s1633 (C=O pyridone), s1562 (CO2asym), s1443, s1375 (CO2symm), s1292, w1184, m819, m756, m686, m517 [IR assignments following Llin\u00e0s et al. (2008To prepare (I) al. 2008].3CO2)2]\u00b7H2O (0.25\u2005mmol) used in place of the manganese acetate tetra\u00adhydrate and the vessel heated to 413\u2005K for 72\u2005h. Upon cooling, green blocks of (II)46H50CuF4N8O12: C 52.80 (52.70), H 4.82 (4.72), N 10.71 (10.64). IR : br3427, br3304, s1633 (C=O pyridone), s1556 (CO2asym), s1435, s1358 (CO2symm), s1294, w1182, w1012, w928, w814, m748, m527.Compound (II)Both (I)Uiso(H) = 1.2\u20131.5Ueq(C) applied. The N- and O-bound H atoms were located in difference maps and refined as riding atoms in their as-found relative positions.Crystal data, data collection and structure refinement details for (I)10.1107/S205698901502424X/sj5492sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S205698901502424X/sj5492Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S205698901502424X/sj5492IIsup3.hklStructure factors: contains datablock(s) II. DOI: 932077, 932078CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The central cyclo\u00adhexane ring has a chair conformation. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming layers parallel to (100). 28H22O5S3, the central cyclo\u00adhexane ring adopts a chair conformation. The atoms of the furan ring attached to the 6-position of the central cyclo\u00adhexane ring are disordered over two sets of sites with occupancies of 0.832\u2005(5) and 0.168\u2005(5). The hy\u00addroxy group is disordered over two positions (at the 4- and 6-positions of the cyclo\u00adhexane ring) in the ratio 0.832\u2005(5):0.168\u2005(5). In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming layers parallel to (100).In the title compound, C The resulting product is a racemate crystallizing in a centrosymmetric space group.Domino or cascade reactions have many applications in organic chemistry (Tietze QT = 0.586\u2005(3)\u2005\u00c5, \u03b8 = 0.0\u2005(3)\u00b0 and \u03c6 = 169\u2005(17)\u00b0]. The mean plane of this ring makes dihedral angles of 80.42\u2005(14), 59.57\u2005(17), 85.65\u2005(17), 66.82\u2005(19), 84.88\u2005(18) and 83.1\u2005(8)\u00b0, respectively, with the five associated five-membered rings .In the title compound, Fig.\u00a01a- and b-axis directions, Figs. and 3. Short S3\u22efS3ii contacts at 3.5210\u2005(12)\u2005\u00c5 may also contribute to the crystal packing method -2,6-bis\u00ad(furan-2-yl)-4-hy\u00addroxy-4-(thio\u00adphen-2-yl)cyclo\u00adhexane-1,3-di\u00adyl]bis\u00ad(thio\u00adphen-2-yl\u00admethanone) was synthesized according to a literature method were found from a difference Fourier map and their positions were constrained to the expected geometries [C\u2014H = 0.95\u00b10.02\u2005\u00c5] with a fixed U value of 0.05\u2005\u00c52. All other H atoms were placed in calculated positions and refined using a riding model with Uiso(H) = 1.2Ueq(carrier).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016009452/sj5500sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989016009452/sj5500Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016009452/sj5500Isup3.cmlSupporting information file. DOI: 1484675CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In this article we report a synthetic procedure and structure of the novel dinuclear copper(II) complex, with a bridging oxalodi\u00adhydroxamate ligand and terminal 2,2\u2032-bi\u00adpyridine and DMSO ligands completing the square pyramidal coordination spheres of the Cu(II) centres.. 2(C2H2N2O4)(C10H8N2)2(C2H6OS)2](ClO4)2, contains two copper(II) ions, connected through an N-deprotonated oxalodi\u00adhydroxamic acid dianion, two terminal 2,2\u2032-bi\u00adpyridine ligands, and two apically coordinating dimethylsulfoxide mol\u00adecules. Two non-coordinating perchlorate anions assure electrical neutrality. The copper(II) ions in the complex dication [Cu2(C10H8N2)2(\u03bc-C2H2N2O4)(C2H6SO)2]2+ are in an O2N3 square-pyramidal donor environment, the Cu\u2013Cu separation being 5.2949\u2005(4)\u2005\u00c5. Two hydroxamate groups in the deprotonated oxalodi\u00adhydroxamic acid are located trans to one each other. In the crystal, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the complex cations to the perchlorate anions. Further C\u2014H\u22efO hydrogen bonds combine with \u03c0\u2013\u03c0 contacts with a centroid-to-centroid separation of 3.6371\u2005(12)\u2005\u00c5 to stack the mol\u00adecules along the a-axis direction.The centrosymmetric binuclear complex, [Cu The twoet al., 2000et al., 2003et al., 2004et al., 2000et al., 1999The C\u2014N and C\u2014C bond lengths in the 2,2\u2032-bi\u00adpyridine ligands are also normal for 2-substituted pyridine derivatives \u2005\u00c5 to stack the cations along the a-axis direction, Fig.\u00a02In the crystal structure, O5\u2014H5trans-form. The bond lengths in oxalodi\u00adhydroxamic acid itself and in its ammonium and thallium salts do not differ significantly [C\u2014C bonds are in the range 1.51\u2005(2)\u20131.528\u2005(3)\u2005\u00c5, C=O 1.231\u2005(3)\u20131.248\u2005(3)\u2005\u00c5, C\u2014N 1.310\u2005(4)\u20131.33\u2005(2)\u2005\u00c5 while the N\u2014O bond lengths vary from 1.36\u2005(2) to 1.388\u2005(1)\u2005\u00c5; Lowe-Ma & Decker, 1986et al., 1987et al., 1991II complexes with ligands derived from doubly or triply deprotonated oxalodi\u00adhydroxamic acid. In one of these complexes 2\u00b76H2O in 10\u2005ml of DMSO the solution of 2,2\u2032-bi\u00adpyridine in 10\u2005ml of methanol was added upon stirring. The resulted solution was stirred for 1\u2005h and then left for slow evaporation.To the warm mixture containing 0.060\u2005g (0.5\u2005mmol) of oxalodi\u00adhydroxamic acid and 0.370\u2005g (1\u2005mmol) of Cu of the title compound.Uiso = 1.2\u20131.5 Ueq(parent atom). The highest peak is located 0.99\u2005\u00c5 from atom Cu1 and the deepest hole is located 0.82\u2005\u00c5 from atom Cu1.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016000050/sj5487sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989016000050/sj5487Isup2.hklStructure factors: contains datablock(s) I. DOI: 1445115CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III ion in the title compound is coordinated by six N atoms of three chelating 1,2-cyclo\u00adhexa\u00adnedi\u00adamine (chxn) ligands, displaying a distorted octa\u00adhedral environment. The crystal packing is stabilized by extensive hydrogen-bonding inter\u00adactions between the N\u2014H groups of the chxn ligands, O\u2014H groups or O atoms of the water mol\u00adecules, chloride ions and Cl atoms of the disordered [ZnCl4]2\u2212 anions.The Cr rac-chxn)3][ZnCl4]Cl\u00b73H2O , has been determined from synchrotron data. The CrIII ion is coordinated by six N atoms of three chelating chxn ligands, displaying a slightly distorted octa\u00adhedral coordination environment. The distorted tetra\u00adhedral [ZnCl4]2\u2212 anion, the isolated Cl\u2212 anion and three lattice water mol\u00adecules remain outside the coordination sphere. The Cr\u2014N(chxn) bond lengths are in a narrow range between 2.0737\u2005(12) and 2.0928\u2005(12)\u2005\u00c5; the mean N\u2014Cr\u2014N bite angle is 82.1\u2005(4)\u00b0. The crystal packing is stabilized by hydrogen-bonding inter\u00adactions between the amino groups of the chxn ligands and the water mol\u00adecules as donor groups, and O atoms of the water mol\u00adecules, chloride anions and Cl atoms of the [ZnCl4]2\u2212 anions as acceptor groups, leading to the formation of a three-dimensional network. The [ZnCl4]2\u2212 anion is disordered over two sets of sites with an occupancy ratio of 0.94:0.06.The structure of the title double salt, [Cr( The synthetic procedures, crystal structures and detailed spectroscopic properties of such [Cr(chxn)3]3+ complexes with chloride or nitrate anions have been reported previously 6][ZnCl4]Cl 2(cyclam)][ZnCl4]Cl\u00b7H2O 3][ZnCl4]Cl\u00b73H2O, (I)3]Cl3\u00b72H2O with 98\u2005K synchrotron data to determine the exact composition and coordination geometry of the CrIII ion. The complex crystallizes in the space group Id with eight formula units in a cell of dimensions a = 18.893\u2005(3) and c = 14.069\u2005(3)\u2005\u00c5. The Cr\u2014N(chxn) bond lengths are in the range 2.0723\u2005(19) to 2.0937\u2005(19)\u2005\u00c5, and the N\u2014Cr\u2014N bite angles are in the range 82.53\u2005(7) to 82.69\u2005(10)\u00b0. In comparison with the bond lengths and angles of the structure of this complex determined with 223\u2005K data rac-chxn)3]3+, three lattice water mol\u00adecules, together with one tetra\u00adhedral [ZnCl4]2\u2212 and one isolated Cl\u2212 counter-ion. The nitro\u00adgen atoms of the three 1,2-cyclo\u00adhexa\u00adnedi\u00adamine ligands define a distorted octa\u00adhedral coordination environment around the Cr(III) ion with a mean N\u2014Cr\u2014N bite angle of 82.1\u2005(4)\u00b0. The resulting five-membered chelate rings of chxn ligands have the expected stable gauche conformation. The Cr\u2014N(chxn) bond lengths are in the range 2.0737\u2005(12) to 2.0928\u2005(12)\u2005\u00c5, in good agreement with those determined in [Cr(RR-chxn)3](NO3)3\u00b73H2O 3]Cl3\u00b72H2O 3](NO3)3\u00b73H2O 3]Cl3\u00b72H2O 3][Co(SS-chxn)3]Cl6\u00b74H2O 3]Cl3\u00b72H2O was prepared according to the literature chloride salt suitable for X-ray structural analysis.Commercially available (Aldrich) Uiso(H) values of 1.2 or 1.5Ueq of the parent atoms. The hydrogen atoms of water mol\u00adecules were restrained using DFIX and DANG commands during the least-squares refinement I. DOI: 10.1107/S2056989016005788/wm5284Isup2.hklStructure factors: contains datablock(s) I. DOI: 1472901CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "All complexes adopt the expected square-planar coordination geometry, and the benzo\u00adthia\u00adzole is engaged in bonding to the metal atom through the imine N atom (Pt\u2014N).Four new platinum(II) complexes, [PtBr3(C8H7NS)] (1), tetra\u00adethyl\u00adammonium tri\u00adbromido\u00adplatinate(II), [NEt4][PtBr3(C9H9NOS)] (2), tetra\u00adethyl\u00adammonium tri\u00adbromido\u00adplatinate(II), [NEt4][PtBr3(C10H11NS)] (3), and tetra\u00adethyl\u00adammonium tri\u00adbromido\u00adplatinate(II), [NEt4][PtBr3(C8H6N2O2S)] (4), have been synthesized and structurally characterized by single-crystal X-ray diffraction techniques. These species are precursors of compounds with potential application in cancer chemotherapy. All four platinum(II) complexes adopt the expected square-planar coordination geometry, and the benzo\u00adthia\u00adzole ligand is engaged in bonding to the metal atom through the imine N atom (Pt\u2014N). The Pt\u2014N bond lengths are normal: 2.035\u2005(5), 2.025\u2005(4), 2.027\u2005(5) and 2.041\u2005(4)\u2005\u00c5 for complexes 1, 2, 3 and 4, respectively. The benzo\u00adthia\u00adzole ligands are positioned out of the square plane, with dihedral angles ranging from 76.4\u2005(4) to 88.1\u2005(4)\u00b0. The NEt4 cation in 3 is disordered with 0.57/0.43 occupancies.Four new platinum(II) complexes, namely tetra\u00adethyl\u00adammonium tri\u00adbromido\u00ad(2-methyl-1,3-benzo\u00adthia\u00adzole-\u03ba All complexes showed the benzo\u00adthia\u00adzoles to coordinate the PtII atom through the imino nitro\u00adgen atom. Also, the benzo\u00adthia\u00adzole is positioned out of the square plane with dihedral angles between 76.4\u2005(4) and 88.1\u2005(4)\u00b0, as previously reported in other platinum\u2013benzo\u00adthia\u00adzole complexes. Given that benzo\u00adthia\u00adzoles have anti\u00adcancer properties, these platinum complexes may have enhanced properties as a result of potential synergism between the ligand and PtII. This deserves further studies as suggested by Noolvi et al. complexes with benzo\u00adthia\u00adzole ligands of general formula (1) crystallizes in an ortho\u00adrhom\u00adbic unit cell with eight formula units. It is a square-planar complex with Pt\u2014N and average Pt\u2014Br bond lengths of 2.035\u2005(5) and 2.433\u2005(6)\u2005\u00c5, respectively, which are within the expected range for PtII complexes. There is no trans-influence observed in the Pt\u2014Br bond trans to the Pt\u2014N bond. The benzo\u00adthia\u00adzole ligand is planar and the methyl group resides in the ligand plane. The dihedral angle between the PtBr3N unit and the benzo\u00adthia\u00adzole ring is 88.1\u2005(4)\u00b0, similar to those observed in other PtII\u2013benzo\u00adthia\u00adzole complexes, as a result of reducing the steric strain between PtBr3 and the benzo\u00adthia\u00adzole ligand ] (2), [NEt4][PtBr3] (3) and [NEt4][PtBr3(5-NO2-2-Me-benzo\u00adthia\u00adzole)] (4) crystallize in the same type of unit cell and space group, monoclinic P21/n, containing four formula units. The Pt\u2014N and average Pt\u2014Br bond lengths for 2, 3, and 4 are 2.025\u2005(4)/2.430\u2005(6)\u2005\u00c5, 2.027\u2005(5)/2.425\u2005(6)\u2005\u00c5 and 2.041\u2005(4)/2.431\u2005(8)\u2005\u00c5, respectively, which are within the expected range. The dihedral angle between PtBr3N and the benzo\u00adthia\u00adzole in 2 is 86.7\u2005(3)\u00b0 and the torsion angle between the aromatic ring and the OCH3 group is 11.9\u2005(7)\u00b0. The C\u2014O (OCH3) bond length is 1.427\u2005(7)\u2005\u00c5, and the C\u2014O\u2014CH3 angle is 116.3\u2005(5)\u00b0. In contrast to 1 and 2, [NEt4][PtBr3] and [NEt4][PtBr3(5-NO2-2-Me-benzo\u00adthia\u00adzole)] have lower dihedral angles between the PtBr3N unit and the benzo\u00adthia\u00adzole ring, 78.6\u2005(4) and 76.(4)\u00b0, respectively. The methyl groups on 3 and 4 are almost co-planar with the benzo\u00adthia\u00adzole plane with deviations \u2264 1.60\u00b0 but in 4, the NO2 group is out of the benzo\u00adthia\u00adzole plane with a torsion angle of 7.5\u2005(7)\u00b0. The C\u2014NO2 bond length is 1.476\u2005(7)\u2005\u00c5, and the O\u2014N\u2014O angle is 117.4\u2005(5)\u00b0. The C\u2014NO2 bond length and O\u2014N\u2014O angle in 4 are smaller than those observed in nitro\u00adbenzene [C\u2014NO2 = 1.486\u2005(2)\u2005\u00c5 and O\u2014N\u2014O = 123.9\u2005(5)\u00b0], which suggests higher electron delocal\u00adization between the nitro group and the aromatic ring in 4 ]\u2212 anions. The [NEt4][PtBr3(2-Me-benzo\u00adthia\u00adzole)] and [NEt4][PtBr3(6-OMe-2-Me-benzo\u00adthia\u00adzole)] complexes showed partial \u03c0-stacking between the phenyl and the thia\u00adzole rings and the corresponding amount of ligand was added with stirring. For 2-methyl-1,3-benzo\u00adthia\u00adzole (99%) 18\u2005\u03bcL were added; for 2-methyl-5-nitro-1,3-benzo\u00adthia\u00adzole (98%) were added, and for 2-methyl-6-meth\u00adoxy-1,3-benzo\u00adthia\u00adzole (97%) were added. The reaction mixtures were stirred without heating until the volume reduced considerably; then the samples were placed in desiccators containing CaCl2 at room temperature to evaporate slowly, leading to the formation of X-ray quality single crystals. For the synthesis with 2,5,6-trimethyl-1,3-benzo\u00adthia\u00adzole (99%), the ligand was added to 20\u2005mL of an acetone solution of [NEt4]2[Pt2Br6] with stirring, and a portion of the reaction mixture was slowly evaporated at 277\u2005K in a small beaker in a secondary container which also contained CaCl2 to form X-ray quality single crystals.Acetone solutions of [NEtd(C\u2014H) = 0.95\u2005\u00c5, Uiso(H) = 1.2Ueq(C); d(C\u2014H2) = 0.99\u2005\u00c5, Uiso(H) = 1.2Ueq(C); d(C\u2014H3) = 0.98\u2005\u00c5, Uiso(H) = 1.5Ueq(C). The NEt4 cation in 3 presented disorder with 0.57/0.43 occupancies.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016002826/bg2580sup1.cifCrystal structure: contains datablock(s) 1, 2, 3, 4. DOI: 10.1107/S2056989016002826/bg25801sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989016002826/bg25802sup3.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S2056989016002826/bg25803sup4.hklStructure factors: contains datablock(s) 3. DOI: 10.1107/S2056989016002826/bg25804sup5.hklStructure factors: contains datablock(s) 4. DOI: 1441324, 1441327, 1441326, 1441325CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I complex, the AgI atom is coordinated by two S atoms of thio\u00adurea and two P atoms of tri\u00adphenyl\u00adphosphane in a distorted tetra\u00adhedral geometry. In the crystal, the component ions are linked by C\u2014H\u22efO, C\u2014H\u22efS, N\u2014H\u22efO and N\u2014H\u22efS hydrogen bonds, generating (10-1) sheets.In the nitrate salt of this Ag 4N2S)2(PPh3)2]NO3, the AgI atom is coordinated by two thio\u00adurea S atoms and two tri\u00adphenyl\u00adphosphane P atoms in a distorted tetra\u00adhedral geometry, with bond angles in the range 102.90\u2005(4)\u2013123.29\u2005(4)\u00b0. The Ag\u2014S=C bond angles are 101.75\u2005(19) and 111.29\u2005(18)\u00b0. In the crystal, the component ions are linked by C\u2014H\u22efO, C\u2014H\u22efS, N\u2014H\u22efO and N\u2014H\u22efS hydrogen bonds, generating (10-1) sheets.In the title salt, [Ag(CH In the cationic complex, [Ag(PPh3)2(tu)2]+, the silver(I) atom is bound to two P atoms of PPh3 and two sulfur atoms of thio\u00adurea, assuming a slightly distorted tetra\u00adhedral geometry , B (C7\u2013C12), C (C13\u2013C18), D (C19\u2013C24), E (C25\u2013C30) and F (C31\u2013C36) are as follows: A/B, A/C, B/C, D/E, D/F and E/F = 82.67\u2005(15), 62.77\u2005(17), 86.59\u2005(14), 73.72\u2005(14), 85.01\u2005(16) and 84.06\u2005(17)\u00b0, respectively. The thio\u00adurea units G (S1/C37/N1/N2) and H (S2/C38/N3/N4) are almost planar with r.m.s. deviations of 0.0031 and 0.0007\u2005\u00c5, respectively, and are oriented at a dihedral angle of 76.82\u2005(11)\u00b0 to each other.In (I)S(6) and In the asymmetric unit, strong N\u2014H\u22efS, N\u2014H\u22efO hydrogen bonds complete distorted s Table\u00a02 and lead3 and PPh3 in a methanol\u2013aceto\u00adnitrile medium . Mixing resulted in the formation of a white precipitate. After stirring for half an hour, the mixture was filtered and the filtrate was left for crystallization. Colorless crystals of (I)et al., 2010The title complex was prepared by adding one equivalent of thio\u00adurea dissolved in 10\u2005ml methanol to a 1:1 mixture of AgNOUiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015001395/hb7352sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015001395/hb7352Isup2.hklStructure factors: contains datablock(s) I. DOI: 1044766CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(9) ring motif. In the crystal, mol\u00adecules are linked via C\u2014H\u22efO hydrogen bonds forming inversion dimers with an R44(10) ring motif. The dimers are linked by C\u2014H\u22efN hydrogen bonds, forming ribbons along [01-1].In the title compound, there is an intra\u00admolecular O\u2014H\u22efN hydrogen bond forming an 22H19ClN4O, the quinolinol moiety is almost planar [r.m.s. deviation = 0.012\u2005\u00c5]. There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond involving the hy\u00addroxy group and a pyridine N atom forming an S(9) ring motif. The dihedral angles between the planes of the quinolinol moiety and the pyridine rings are 44.15\u2005(9) and 36.85\u2005(9)\u00b0. In the crystal, mol\u00adecules are linked via C\u2014H\u22efO hydrogen bonds forming inversion dimers with an R44(10) ring motif. The dimers are linked by C\u2014H\u22efN hydrogen bonds, forming ribbons along [01-1]. The ribbons are linked by C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions [inter-centroid distance = 3.7109\u2005(11)\u2005\u00c5], forming layers parallel to (01-1).In the title compound, C Analysis calculated for C22H19ClN4O: C 67.60, H 4.90, N 14.33%; found: C 67.50, H 5.01, N 14.37%.A suspension of paraformaldehyde and bis\u00ad(2-pyridyl\u00admeth\u00adyl)amine in 100\u2005ml of MeOH was stirred for 18\u2005h at room temperature. The solvent was removed under vacuum. To the product obtained was added 100\u2005ml of toluene and 5-chloro-8-quinolinol , and the mixture was heated for 24\u2005h at 353\u2005K. The solvent was removed under vacuum to give an oily product, which was crystallized from hexa\u00adne\u2013di\u00adchloro\u00admethane. The crude solid was recrystallized from aceto\u00adnitrile to obtain yellow crystals of the title compound . HRMS (Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015022410/su5241sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015022410/su5241Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015022410/su5241Isup3.cmlSupporting information file. DOI: 1438483CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II coordination polymer has been prepared via solvothermal synthesis using dimethyl sulfoxide as solvent. The coordination polymer forms double-chains along [010] and exhibits \u03c0\u2013\u03c0 stacking and C\u2014H\u22ef\u03c0 inter\u00adactions forming the inter\u00adior of the double-chains, separated from a C\u2014H\u22ef\u03c0 hydrogen-bonding network in the space between the double-chains.A one-dimensional Ni 7H3NO4)(C4H4N2)(C2H6OS)]n, consists of [010] chains composed of NiII ions linked by bis-monodentate-bridging pyrazine mol\u00adecules. Each of the two crystallographically distinct NiII ions is located on a mirror plane and is additionally coordinated by a dimethyl sulfoxide (DMSO) ligand through the oxygen atom and by a tridentate 2,6-pyridine-di\u00adcarb\u00adoxy\u00adlic acid dianion through one of each of the carboxyl\u00adate oxygen atoms and the pyridine nitro\u00adgen atom, leading to a distorted octa\u00adhedral coordination environment. The title structure exhibits an inter\u00adesting complementarity between coordinative bonding and \u03c0\u2013\u03c0 stacking where the Ni\u2014Ni distance of 7.0296\u2005(4)\u2005\u00c5 across bridging pyrazine ligands allows the pyridine moieties on two adjacent chains to inter\u00addigitate at halfway of the Ni\u2014Ni distance, resulting in \u03c0\u2013\u03c0 stacking between pyridine moieties with a centroid-to-plane distance of 3.5148\u2005(2)\u2005\u00c5. The double-chain thus formed also exhibits C\u2014H\u22ef\u03c0 inter\u00adactions between pyridine C\u2014H groups on one chain and pyrazine mol\u00adecules on the other chain. As a result, the inter\u00adior of the double-chain structure is dominated by \u03c0\u2013\u03c0 stacking and C\u2014H\u22ef \u03c0 inter\u00adactions, while the space between the double-chains is occupied by a C\u2014H\u22efO hydrogen-bonding network involving DMSO ligands and carboxyl\u00adate groups located on the exterior of the double-chains. This separation of dissimilar inter\u00adactions in the inter\u00adior and exterior of the double-chains further stabilizes the crystal structure.The title coordination polymer, [Ni(C II complex obtained by reacting 2,6-pyridine di\u00adcarb\u00adoxy\u00adlic acid and nickel carbonate using water as solvent , where each of the NiII atoms is coordinated by a 2,6-pyridine-di\u00adcarb\u00adoxy\u00adlic acid dianion, a pyrazine mol\u00adecule, and a DMSO ligand \u2005\u00c5, i.e. the length of the b axis.The asymmetric unit contains two half Nind Fig.\u00a01. The triII chains form a double-chain structure via \u03c0\u2013\u03c0 stacking between their pyridine moieties . Double-chains of mol\u00adecule A are linked by C21A\u2014H21E\u22efO2Ai/iv, C12A\u2014H12A\u22efO1Ai, C21A\u2014H21D\u22efO4Aiii, and C22A\u2014H22D\u22efO4Aiii hydrogen bonds to form sheets extending along the same direction. Thus, alternating sheets with an ABAB pattern can be observed. Two neighboring sheets are connected via C11A\u2014H11A\u22efO5B and C11B\u2014H11B\u22efO5A hydrogen bonds to form a three-dimensional network. The hydrogen-bond lengths and angles are summarized in Table\u00a01In contrast to the \u03c0\u2013\u03c0 stacking and C\u2014H\u22ef\u03c0 inter\u00adactions forming the inter\u00adior of the double-chains, the exterior of the double-chains is mainly occupied by polar DMSO mol\u00adecules and carboxyl\u00adate groups. As a result, a network of C\u2014H\u22efO hydrogen bonds exists in the space between the double-chains Fig.\u00a03, linkingIn summary, a separation of dissimilar inter\u00adactions can be observed between the non-covalent lipophilic \u03c0\u2013\u03c0 stacking and C\u2014H\u22ef\u03c0 inter\u00adactions in the inter\u00adior of the double-chains and the polar hydrogen bonds in the exterior of the double-chains, further stabilizing the crystal structure.3 , 2,6-pyridine di\u00adcarb\u00adoxy\u00adlic acid , and pyrazine were dissolved in 10\u2005ml dimethyl sulfoxide. The resulting mixture was transferred into a stainless steel autoclave which was heated at 373\u2005K for 24\u2005h and cooled to room temperature at a cooling rate of 0.1\u2005K per minute. Green needle-like crystals of the title compound were collected by filtration. Selected IR bands : 1640.6 (C=O), 1367.9 (C\u2014O), 950.9 (S=O), 480.6 (bridging pyrazine).Anhydrous NiCOUiso(H)= 1.2/1.5Ueq(C). Methyl H atoms were allowed to rotate around the corresponding C\u2014C bond. There are two disordered parts, both of which are in mol\u00adecule A. The carboxyl\u00adate atom O2A sits just outside of the mirror plane (occupancy 0.5) and one of the DMSO methyl groups is disordered over two positions in a ratio of 0.54\u2005(2):0.46\u2005(2). The C atom of this group was refined with isotropic displacement parameters.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016007064/wm5288sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016007064/wm5288Isup2.hklStructure factors: contains datablock(s) I. DOI: 1476677CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "We have evaluated the efficacy of dapagliflozin in patients with type 1 diabetes mellitus (DM1) without adequate control. We expected that adding dapagliflozin to this population on top of their base treatment would lower their HbA1c levels.We conducted a pragmatic, open, 24-week study of treatment with 10\u00a0mg of oral dapagliflozin in patients with DM1 and chronic hyperglycemia. We evaluated glycemic control, lipid profile, weight, and insulin dose. Safety was assessed by adverse event reporting.p\u2009=\u20090.08). The hemoglobin A1C (HbA1C) level decreased from 9.18\u2009\u00b1\u20091.02 (77\u2009\u00b1\u200911.1\u00a0mmol/mol) to 8.05\u2009\u00b1\u20091.09\u00a0% (64\u2009\u00b1\u200911.9\u00a0mmol/mol) (p\u2009=\u20090.0156); total cholesterol decreased from 299\u2009\u00b1\u200912 to 199\u2009\u00b1\u20097\u00a0mg/dL (p\u2009=\u20090.02); triglycerides decreased from 184\u2009\u00b1\u200915 to 160\u2009\u00b1\u200911\u00a0mg/dL (p\u2009=\u20090.0002), HDL-C decreased from 40\u2009\u00b1\u200917 to 42\u2009\u00b1\u20099\u00a0mg/dL (p\u2009=\u20090.54); and LDL-C decreased from 187\u2009\u00b1\u200919 to 170\u2009\u00b1\u200921\u00a0mg/dL (p\u2009=\u20090.049). No adverse events were reported.Fasting glucose levels decreased from 176.42\u2009\u00b1\u200945.33\u00a0mg/dL to 139.67\u2009\u00b1\u200944.42\u00a0mg/dL (p\u2009=\u20090.05); although no significant valued was reached, postprandial glucose showed a decreased tendency from 230.25\u2009\u00b1\u200952.06\u00a0mg/dL to 193.83\u2009\u00b1\u200945.43\u00a0mg/dL (The beneficial effects of SGLT2 inhibitors on metabolic control and their safety after a 24-week open study demonstrate their potential indication as an adjunctive treatment with insulin in patients with DM1; however, long-term clinical trials should be considered. Treatment of Type 1 diabetes mellitus (DM1) is presently restricted to insulin and in selected cases, pramlintide and islet or pancreas transplantation .In contrast the management of DM1 with oral drugs is limited. Sodium/glucose cotransporter 2 inhibitors (SGLT2I), as initial or in combination therapy have recently been used to treat type 2 diabetes mellitus (DM2) . These dThe aim of this study was to evaluate the efficacy of dapagliflozin in a group of patients with poorly controlled DM1. We expected that adding dapagliflozin to this population\u2019s insulin regimen, would improve their glycemic control.We performed an open, 24-week, pragmatic clinical trial in a group of patients with poorly controlled DM1 in a private clinic in Monterrey, Nuevo Leon, M\u00e9xico from 2013\u20132014. Twelve patients met the following inclusion criteria: DM1 according to the American Diabetes Association classification ; both geChanges in fasting and postprandial glucose and HbA1C were considered to be the primary outcomes. Appearance of hypoglycemia, genitourinary or micotic infections and ketonuria were routinely revised.The study protocol was approved by the local research ethics committee.2.Twelve patients were included with a mean age of 27\u2009\u00b1\u200911\u00a0years and a mean duration of disease of 9.17\u2009\u00b1\u20097.41\u00a0years. All were overweight with a body mass index (BMI) of 27.98\u2009\u00b1\u20091.97\u00a0kg/mp\u2009=\u20090.05). The postprandial glucose decreased from 230.25\u2009\u00b1\u200952.06\u00a0mg/dL to 193.83\u2009\u00b1\u200945.43\u00a0mg/dL; however, it did not reach a significant value (p\u2009=\u20090.08). HbA1C significally decreased from 9.18\u2009\u00b1\u20091.02\u00a0% (77\u2009\u00b1\u200911.1\u00a0mmol/mol) to 8.05\u2009\u00b1\u20091.09\u00a0% (64\u2009\u00b1\u200911.9\u00a0mmol/mol) (p\u2009=\u20090.0156). The lipid profile was modified favorably, with total cholesterol decreasing from 299\u2009\u00b1\u200912 to 199\u2009\u00b1\u20097\u00a0mg/dL (p\u2009=\u20090.02), LDL-C from 187\u2009\u00b1\u200919 to 170\u2009\u00b1\u200921\u00a0mg/dL (p\u2009=\u20090.049) and triglycerides from 184\u2009\u00b1\u200915 to 160\u2009\u00b1\u200911\u00a0mg/dL (p\u2009=\u20090.0002). HDL-C was unchanged at 40\u2009\u00b1\u200917 and 42\u2009\u00b1\u20099\u00a0mg/dL (p\u2009=\u20090.54) [\u00a0weeks [8The lipid profile also improved following treatment, with reductions in total cholesterol, c-LDL cholesterol and triglycerides . This is inconsistent with previous SGLT-2 studies and may have resulted from improved glycemic control, weight loss, or other causes , 9.An unattained metabolic control of DM1 has prompted a search for other therapeutic options, like pramlintide or islets transplantation , 10\u201312.Evidence of efficacy of SGLT2I in DM1 is limited ; howeverWe believe that dapagliflozin is safe and well tolerated with no clinically documented adverse effects. In addition, our study had the strength of a pragmatic design with a relatively long treatment period.However, recently the U.S. Food and Drug Administration (FDA) issued a Drug Safety Communication warning of an increased risk of diabetic ketoacidosis associated with the used of all the approved SGLT2I. This potential complication related is predictable, detectable, and preventable, with the full picture still favoring the use of SGLT2I DM1 patient .Nevertheless, we recognize that the data should be cautiously considered because the study was performed in a specialized care center with educated patients and a small sample size of overweight individuals. Also, we do not have information regarding the follow up of the patients.Dapagliflozin as an adjunct therapy to insulin caused significant changes in the HbA1C level, fasting and postprandial glucose and atherogenic lipid in patients with DM1. Thus, dapagliflozin may represent a new therapeutic approach, although long-term controlled clinical trials with a greater number of patients are needed to confirm our data."} +{"text": "N-{4-[(6-chloro\u00adpyridin-3-yl)meth\u00adoxy]phen\u00adyl}-2,6-di\u00adfluoro\u00adbenzamide is reported. The crystal packing is stabilized by N\u2014H\u22efN, C\u2014H\u22efO, C\u2014H\u22efF and C\u2014H\u22ef\u03c0 hydrogen bonds supplemented by offset \u03c0\u2013\u03c0 stacking inter\u00adactions.The mol\u00adecular and crystal structure of 19H13ClF2N2O2, the conformation of the N\u2014H bond in the amide segment is anti to the C=O bond. The mol\u00adecule is not planar, with dihedral angles between the central benzene ring and the outer benzene and pyridyl rings of 73.35\u2005(7) and 81.26\u2005(6)\u00b0, respectively. A weak intra\u00admolecular C\u2014H\u22efO hydrogen bond occurs. In the crystal, N\u2014H\u22efN, C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds lead to the formation of dimers. The N\u2014H\u22efN inversion dimers are linked by \u03c0\u2013\u03c0 contacts between adjacent pyridine rings [centroid\u2013centroid = 3.8541\u2005(12)\u2005\u00c5] and C\u2014H\u22ef\u03c0 inter\u00adactions. These contacts combine to stack the mol\u00adecules along the a axis.In the title compound, C The mixture was stirred at 283\u2013288\u2005K for 2\u2005h, then washed with 0.5% hydro\u00adchloric acid solution, and a saturated aqueous solution of sodium hydrogen carbonate, dried and evaporated. The residue was recrystallized from di\u00adchloro\u00admethane, giving colourless blocks of the title compound after three weeks.N-(4-(pyridin-3-ylmeth\u00adoxy)phen\u00adyl)benzamide or its substituted derivatives gave no hits. However, structures of eight substituted 2,6-di\u00adfluoro-N-phenyl\u00adbenzamide derivatives were found, see for example Cockroft et al. = 0.93\u20130.97\u2005\u00c5 and Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015023701/sj5486sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015023701/sj5486Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015023701/sj5486Isup3.cmlSupporting information file. DOI: 1441555CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The position of the second non-linking cage C atom was established unambiguously by geometric and crystallographic methods and there is no evidence of C/B disorder.In 1,1\u2032-bis[1,7-dicarba- 4H22B20, the two {1,7-closo-C2B10H11} cages are linked across a centre of inversion, with C\u2014C = 1.5401\u2005(16)\u2005\u00c5. The position of the second non-linking cage C atom was established unambiguously by geometric and crystallographic methods and there is no evidence of C/B disorder.In the title compound, C There are several isomeric possibilities for bis\u00ad(carboranes) composed of two C2B10 icosa\u00adhedra. Bis(ortho-carborane), 1,1\u2032-bis\u00ad , 1,1\u2032-bis, the subject of this study is, however, less well known. It was first prepared by Zakharkin & Kovredov ], (I)Whilst the chemistry of single-cage carboranes is well developed linked across a crystallographic inversion centre by the C1\u2014C1A bond \u00b0, and the facing penta\u00adgons B2/B3/B4/B5/B6 and B2A/B3A/B4A/B5A/B6A are staggered. The five C1\u2014B distances span the range 1.7107\u2005(12)\u20131.7385\u2005(12)\u2005\u00c5, whilst C7\u2014B connectivities lie between 1.6967\u2005(13) and 1.7180\u2005(13)\u2005\u00c5, with, in both cases, the two shortest distances being to the B atoms (B2 and B3) that lie between the C atoms. The B2\u2014B3 connectivity, 1.7947\u2005(13)\u2005\u00c5, is the longest B\u2014B link, with all (19) others lying between 1.7709\u2005(13) and 1.7891\u2005(15)\u2005\u00c5. In general terms these C\u2014B and B\u2014B distances are fully consistent with the averages recently calculated, 1.705\u2005(14) and 1.772\u2005(11)\u2005\u00c5, respectively z] Fig.\u00a01. The twoB at 2.39\u2005\u00c5 . Although CH units and BH units in carboranes are protonic and hydridic, respectively, there is no evidence of di\u00adhydrogen bonding, the shortest such contact being H7\u22efH12C at 2.61\u2005\u00c5 [symmetry code: (C) \u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a0The only H\u22efH contact less than 2.40\u2005\u00c5 is H6\u22efH6closo-C2B10 fragment using Conquest (Version 1.16) returned 132 hits of which only two involve the 1,1\u2032-bis\u00ad unit. In DUWJAH 2} groups attached to C7 and C7\u2032 whilst in DUWJEL (OMe)} units. Of the remaining 130 hits there are five cases of the parent mol\u00adecule 1,7-closo-C2B10H12 co-crystallized with other mol\u00adecules, the first of these to be reported being the hexa\u00admethyl\u00adphospho\u00adr\u00adamide adduct TOKGOP was refined and then analysed by both the Vertex-to-Centroid Distance = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S1600536814022132/hb7290sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814022132/hb7290Isup2.hklStructure factors: contains datablock(s) I. DOI: 1027936CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H = (1\u20133/Nr)In the Methods section, Equation 2b is incorrect. The correct equation is: \u03b8"} +{"text": "I ions complexed by ferrocenediyl-1,1\u2032-dicarbo\u00adnitrile forming a paddle-wheel with two acetone mol\u00adecules, with one coordinating on top of one trigonal\u2013planar-coordinated copper ion, and the other as a packing solvent.The first ferrocenylcarbo\u00adnitrile copper complex is reported. The structure consists of two Cu O)tris\u00addicopper(I) bis\u00ad(tetra\u00adfluorido\u00adborate) acetone monosolvate, [Cu2Fe3(C6H4N)6(C3H6O)](BF4)2\u00b7C3H6O, consists of two CuI ions bridged by a ferrocene-1,1\u2032-dicarbo\u00adnitrile moiety in a paddle-wheel-architectured sandwich complex with two BF4\u2212 units as counter-ions. One of the latter is equally disordered over two sets of sites. The two CuI ions are complexed in a trigonal\u2013planar manner by three nitrile N-donor atoms. Further inter\u00adactions by the O atom of an acetone mol\u00adecule to one of the CuI atoms and a weak \u03b72,\u03c0-inter\u00adaction of two atoms of a cyclo\u00adpenta\u00addienyl ring to the other CuI atom complete a distorted trigonal\u2013pyramidal environment for each of the metal ions. A further acetone mol\u00adecule is also present as a solvent mol\u00adecule. The crystal packing is consolidated by several \u03c0\u2013\u03c0 inter\u00adactions.The mol\u00adecular structure of (acetone-\u03ba M\u2014C\u00a0C\u2014 , showing moderate electron communication between two redox-active metallocenyl termini in the mixed-valence species \u2013phenyl\u00adene bridge shows a moderate delocalization. In contrast, a weak electron transfer by generation of the mixed-valence species [Ru(N\u00a0CFc)(NH3)5]3+ [Fc = Fe(\u03b75-C5H4)(\u03b75-C5H5)] has been described bridging unit between two redox-active ferrocenyl moieties 2CO}{\u03bc-Fe(\u03b75-C5H4C\u00a0N)2}3](BF4)2\u00b7(CH3)2CO, (I)The electron-transfer properties of the acetyl\u00adide function have been investigated intensively by using bridging units of the type \u2014C\u00a0C\u2014 al. 2011 describe2Fe3 complex mol\u00adecule in the asymmetric unit consisting of two CuI ions bridged by three 1,1\u2032-ferrocenediyl dicarbo\u00adnitrile ligands that form a triangular paddle-wheel sandwich-type complex with iron\u22efiron distances ranging from 9.1739\u2005(13) (Fe2\u22efFe3) to 10.0385\u2005(12)\u2005\u00c5 . The complex crystallizes with two BF4\u2212 counter-ions and two mol\u00adecules of acetone. One acetone mol\u00adecule coordinates with its oxygen atom to Cu1 [Cu1\u2014O1 2.375\u2005(2)\u2005\u00c5], leading to an 18 VE complex and an overall distorted trigonal\u2013pyramidal environment. The Cu2 ion exhibits a weak inter\u00admolecular \u03b72, \u03c0 inter\u00adaction as compared to Cu2 [0.0602\u2005(16)\u2005\u00c5] due to a stronger inter\u00adaction with the axial moiety. The Cu\u22efCu distance [3.3818\u2005(7)\u2005\u00c5] exceeds the sum of the van der Waals radii \u00b0; \u03b1 C5\u22efCu2, 93.23\u2005(1) \u00b0; Table\u00a01Besides the already noted inter\u00admolecular inter\u00adaction between Cu2 and the mid-point of the C26\u2014C27 bond, \u03c0\u2013\u03c0 inter\u00adactions are present in the crystal packing between the C23 atom and its symmetry-generated equivalent (BF4)2\u00b7(CH3)2CO: Copper powder , Cu(BF4)2\u00b75H2O and ferrocene-1,1\u2032-dicarbo\u00adnitrile were stirred in 5\u2005ml of di\u00adchloro\u00admethane at room temperature overnight. The resulting orange precipitate was filtered off using zeolite and washed several times with 20\u2005ml of di\u00adchloro\u00admethane until the filtrate was colorless. The residue was taken up in acetone and this solution was evaporated to dryness using a rotary evaporator affording (I)4]2\u00b75H2O). IR : \u03bd = 2248 (CN). 1H NMR = 5.12 , 4.82 . 13C{1H} NMR: Data not available due to low solubility. HRMS (ESI\u2013TOF): M+ C12H8N2CuFe (C24H16N4CuFe2): m/z = 534.9342 ; M+ C24H16N4CuFe2 (C12H8N2CuFe): m/z = 298.9342 .Ferrocene-1,1\u2032-dicarbo\u00adnitrile was prepared according to a published procedure (Strehler Uiso(H) = 1.2Ueq(C) and a C\u2014H distance of 0.93\u2005\u00c5 for aromatic and Uiso(H) = 1.5Ueq(C) and a C\u2014H distance of 0.96\u2005\u00c5 for methyl H atoms. The F atoms of one of the two BF4\u2212 ions were refined as equally disordered over two sets of sites using DFIX [B\u2014F 1.38\u2005(2)\u2005\u00c5] and DANG [F\u2014F 2.25\u2005(4)\u2005\u00c5] instructions. Since some anisotropic displacement ellipsoids were rather elongated, DELU/SIMU/ISOR restraints were also applied I. DOI: 10.1107/S2056989015001760/wm5104Isup2.hklStructure factors: contains datablock(s) I. DOI: 1045804CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "AbstractParaplonobia Wainstein and Neopetrobia Wainstein (Trombidiformes: Tetranychidae) are reported for the first time from Saudi Arabia. Three new species Paraplonobia (Anaplonobia) arabica Mirza & Alatawi, sp. n., Paraplonobia (Anaplonobia) haloxylonia Alatawi & Mirza, sp. n. and Paraplonobia (Anaplonobia) tabukensis Kamran & Alatawi, sp. n. are described and illustrated based on adult females, collected from Prosopisjuliflora (SW.) Dc. (Fabaceae) and Haloxylonsalicornicum Bunge (Amaranthaceae) from two different regions of Saudi Arabia. Neopetrobiamcgregori (Pritchard and Baker) is redescribed and illustrated based on female collected from Cynodondactylon L. (Poaceae).The diagnostic morphological features including leg chaetotaxy of all known species of the subgenus Anaplonobia is tabulated. A key to the world species of the genus Paraplonobia is also provided.The two tetranychid genera Paraplonobia Wainstein belongs to the tribe Hystrichonychini Pritchard and Baker of the subfamily Bryobiinae (Acari: Prostigmata: Tetranychidae). Anaplonobia and Paraplonobia as subgenera of Aplonobia Womersley. Later, Anaplonobia and Paraplonobia as two valid genera. After that, Paraplonobia into three subgenera: Anaplonobia Wainstein, Brachynychus Mitrofanov & Strunkova and Paraplonobia Wainstein on the basis of coxal setal count and the aspect of peritremes and considered the genus Anaplonobia as subgenus of Paraplonobia , Eutetranychuspalmatus Attiah, Oligonychusafrasciaticus (McGregor), Oligonychuspratensis (Banks), Tetranychuscinnabarinus , Tetranychusturkrestzni (Ugarov & Nikolskii), and Tetranychusurticae (Koch) harteni Meyer and Paraplonobia (Paraplonobia) dactyloni Smiley & Baker were reported from Yemen (A few tetranychid species have been reported from Saudi Arabia (SA): e (Koch) . The genom Yemen .Paraplonobia and Neopetrobia, are reported upon for the first time from Saudi Arabia with three new species: Paraplonobia (Anaplonobia) arabica sp. n., Paraplonobia (Anaplonobia) haloxylonia sp. n. and Paraplonobia (Anaplonobia) tabukensis sp. n. which are described and illustrated based on adult females. The male of Paraplonobia (Anaplonobia) haloxylonia sp. n. is also described and illustrated. Neopetrobiamcgregori (Pritchard & Baker) is redescribed and illustrated based on female.Two genera, Anaplonobia are provided including body morphological features, leg I length in comparison to body length, and leg chaetotaxy haloxylonia sp. n., female paratype each of Paraplonobia (Anaplonobia) arabica sp. n., and Paraplonobia (Anaplonobia) tabukensis sp. n., with Accession numbers, OSAL 0115769, OSAL 00115768, OSAL 0110333 and OSAL 0110332 respectively, that were deposited at UniversityOhio State Acarology Laboratory(OSAL), USA.The mite specimens were collected by shaking the plant parts, especially leaves, onto a white sheet of paper. Mites found moving on paper were collected with the help of a camel hairbrush and preserved in small vials containing 70% ethanol. Preserved mite specimens were observed under a stereomicroscope and mounted on glass slides in Hoyer\u2019s medium. The mounted specimens were examined under phase contrast microscope . Different body parts were pictured using an auto montage software system , then drawn with Adobe Illustrator . All measurements are in micrometers. The terminology used in this paper follows that of Taxon classificationAnimaliaProstigmataTetranychidaeGenusWainstein, 1960Aplonobia (Paraplonobia) Wainstein, 1960: 140.Paraplonobia : Aplonobia (Paraplonobia) echinopsili Wainstein, 1960 by original designation.Based on Baker and Tuttle 1968, Gutierrez 1955, 1 normal in position, coxal setal formula variable, most species with 2\u20132\u20131\u20131 except one species of the subgenus Brachynychus having 4\u20133\u20132\u20132 setae on coxae I\u2013IV respectively; anal setae three pairs; peritremes simple/anastomosing; tarsus I with two sets of duplex setae, present close to distal end of tarsus; claws and empodium pad-like each with tenant hairs Eight species with dorsal body setae slightly shorter/as long as or longer than distances to the bases of consecutive setae (Tables Paraplonobia (Anaplonobia) arabica sp. n., Paraplonobia (Anaplonobia) haloxylonia sp. n., and Paraplonobia (Anaplonobia) tabukensis sp. n.) reported in this study , comparative length of setae with respect to the distance of setae next behind, shape of peritremes , propodosomal shield , hysterosoma , comparative length of leg I with respect to body length (shorter/longer) and leg chaetotaxy are the major diagnostic characters vary among/within the species of subgenus ia Table , 2.Anaplonobia have been reported from USA, Mexico, South Africa and Pakistan and collected mostly from three host plants families Asteraceaea, Fabaceae and Poaceae (Most species of the subgenus Poaceae (Table 1Paraplonobia (Anaplonobia) arabica sp. n., collected from Prosopisjuliflora from three different regions of Saudi Arabia, are morphologically similar except for some variations in setal counts on Tibia II and Tarsus I\u2013II\u2013III. (Table Paraplonobia (Anaplonobia) prosopis had been found also in the description made by Anaplonobia, setal variations on genua, tibiae and tarsi have been found among the different specimens collected from the same host and location within the same species. i.e. genua I (8\u20139) in Paraplonobia (Anaplonobia) candicans, tibia I (12\u201313) and tarsus II (12\u201313) of Paraplonobia (Anaplonobia) glebulanta, and tarsus III (12\u201313) of Paraplonobia (Anaplonobia) theroni . Measurement of holotype followed by 8 paratypes (in parenthesis) Figs \u20138.Dorsum , sc1 40 (38\u201341), sc2 41 (40\u201343), c1 45 (44\u201348), c2 42 (40\u201344), c3 40 (39\u201344), d1 34 (32\u201338), d2 44 (43\u201346), e1 45 (44\u201348), e2 44 (43\u201345), f1 45 (44\u201345), f2 44 (42\u201345), h1 46 (45\u201348). Distance between dorsal setae: v2\u2013v2 53 (51\u201355), v2\u2013sc1 97 PageBreak(95\u201398), sc1\u2013sc2 56 (54\u201357), sc1\u2013sc1 166 (162\u2013167), sc2\u2013sc2 263 (260\u2013266), c1\u2013c1 94 (92\u201396), c1\u2013c2 82 (80\u201385), c2\u2013c3 41 (39\u201344), c2\u2013c2 261 (260\u2013264), c3\u2013c3 345 (340\u2013346), d1\u2013d1 82 (80\u201384), d1\u2013d2 81 (80\u201382), d2\u2013d2 226 (224\u2013228), c1\u2013d1 75 (74\u201378), c3\u2013d2 97 (95\u201399), e1\u2013e1 63 (61\u201365), e1\u2013e2 78 (74\u201379), e2\u2013d2 85 (83\u201386), e2\u2013e2 79 (75\u201379), f1\u2013f1 63 (60\u201365), f2\u2013f2 107 (105\u2013108), f1\u2013f2 53 (50\u201354), f1\u2013d1 69 (66\u201370), h1\u2013h1 57 (55\u201359).sum Fig. . Body ovVenter , g2 40 (38\u201342), g1\u2013g1 40 (39\u201344), g2\u2013g2 56 (52\u201357), g1\u2013g2 12 (10\u201314); anal setae three pairs, ps1 21 (18\u201324), ps2 37 (35\u201339), ps3 58 (54\u201360), ps1\u2013ps1 33 (30\u201334), ps2\u2013ps2 26 (24\u201327), ps3\u2013ps3 19 (18\u201322); para-anal setae two pairs, h2 33 (31\u201334), h2\u2013 h2 17 (16\u201318), h3 38(35\u201340), h3\u2013h3 46 (45\u201348).ter Fig. . IdiosomGnathosoma , Deesa Valley, Dessa, Tabuk, SA, 27\u00b036.048'N, 036\u00b025.592'E, October, 18, 2015, coll. J.H. Mirza.; seven paratype females, Prosopisjuliflora (Fabaceae), Sharma, Near Red Sea, Tabuk, SA, 28\u00b003.479'N, 035\u00b017.186'E, October, 19, 2015, coll. M. Kamran.Holotype and one paratype female, Paraplonobia (Anaplonobia) arabica sp. n. relates to Paraplonobia (Anaplonobia) prosopis , Paraplonobia (Anaplonobia) algarrobicola and Paraplonobia (Anaplonobia) boutelouae Tuttle & Baker, 1968 because of sharing following similar characters: dorsal body setae spatulate and distinctly shorter to the distances of setae next behind and widely spaced dorsal hysterosomal striations. Also, the new species closely resembles Paraplonobia (Anaplonobia) prosopis by setae c1 at least reaching half distance to the bases of setae d1. However, the new species differs PageBreakfrom all related species by having stylophore anteriorly with slight incision (notch). The new species is also distinguished from Paraplonobia (Anaplonobia) prosopis by setae c1 reaching to the distance of setae d1 (2/3 vs.1/2), setae c1\u2013c1 almost 1.5 times widely spaced than setae f1\u2013f1 vs. almost sub/equally spaced in Paraplonobia (Anaplonobia) prosopis. The new species can be separated from other related species Paraplonobia (Anaplonobia) algarrobicola and Paraplonobia (Anaplonobia) boutelouae by the setae c1 reaching 2/3 to the distance of d1 vs. less than half as long as distances to the bases setae next behind in later speciesThe Taxon classificationAnimaliaProstigmataTetranychidaehttp://zoobank.org/09E8353-E635-4C38-B277-8D6DDC56C31ADorsal setae lanceolate, densely serrate, not set on tubercles and distinctly shorter to the distances of setae next behind, dorsocentral setae almost 1/3 to the distance of setae next behind, propodosoma medially with weak, longitudinal irregular striations, hysterosoma with transverse and closely spacedstriations medially, stylophore slightly notched anteriorly, peritremes anastomosed distally, with few long thread like branches, and hysterosomal striations closely spaced, leg I shorter than body.n = 39). Measurements of holotype followed by 38 paratypes (in parenthesis) , sc1 24 (23\u201325), sc2 22 (21\u201324), c1 19 (18\u201321), c2 22 (21\u201323), c3 25 (24\u201328), d1 15 (12\u201316), d2 18 (17\u201319), e1 16 (15\u201317), e2 20 (19\u201320), f1 25 (24\u201328), f2 31 (29\u201332), h1 34 (32\u201335). Distance between dorsal setae: v2\u2013v2 72 (70\u201373), v2\u2013sc1 75 (72\u201378), sc1\u2013sc2 66 (63\u201367), sc1\u2013sc1 167 (163\u2013172), sc2\u2013sc2 254 (250\u2013259), c1\u2013c1 90 (88\u201392), c1\u2013c2 75 (71\u201378), c2\u2013c3 81 (78\u201385), c2\u2013c2 231 (229\u2013234), c3\u2013c3 373 (372\u2013375), d1\u2013d1 91 (89\u201392), d1\u2013d2 65 (62\u201369), d2\u2013d2 204 (201\u2013206), c1\u2013d1 103 (100\u2013104), c3\u2013d2 160 (158\u2013161), e1\u2013e1 55 (53\u201357), e1\u2013e2 53 (50\u201354), e2\u2013d2 85 (82\u201386), e2\u2013e2 150 (148\u2013152), f1\u2013f1 60 (59\u201362), f2\u2013f2 80 (78\u201383), f1\u2013f2 28 (25\u201329), f1\u2013d1 93 (91\u201394), h1\u2013h1 31 (28\u201332).sum Fig. . Body ovVenter , g2 20 (19\u201321), g1\u2013g1 32 (31\u201333), g2\u2013g2 35 (34\u201336), g1\u2013g2 10 (10\u201312); anal setae three pairs, ps1 11 (10\u201312), ps2PageBreakPageBreakPageBreak16 (15\u201317), ps3 17(16\u201318), ps1\u2013ps1 16 (15\u201318), ps2\u2013ps2 22 (20\u201323), ps3\u2013ps3 26 (25\u201326); para\u2013anal setae two pairs, h2 16 (15\u201317), h2\u2013 h2 14 (13\u201316), h3 17 (15\u201317), h3\u2013h3 31 (30\u201332) Fig. .Gnathosoma \u20139\u20139\u20139; tarsi I with 15 tactile setae, two sets of duplex setae at distal end, 11 tactile setae and two solenidia well proximal to duplex setae, two eupathidia; tarsi II with 10 tactile setae, one set of duplex setae, two eupathidia and one solenidion; tarsi III with 12 tactile setae and one solenidion; tarsi IV with 12 tactile setae and one solenidion. True claws pad like each with one pair of tenant hair; empodium pad-like with two rows of small tenant hairs.egs Figs . Length Male (n = 11) Figs \u201328. Dorssum Fig. . Body alVenter , anal setae three pairs ; para-anal setae two pairs ; aedeagus up turned, broadly sigmoid, sharply tapering distally , Salbookh Road, Dariyah, Riyadh, SA, 24\u00b030.649'N, 46\u00b046.615'E, September, 18, 2012, coll. M. Kamran; four males and 22 female paratypes, Hilaria sp. (Poaceae), Tashlia, Heyer Road, Riyadh, SA, 24\u00b029.000'N, 46\u00b047.890'E, January, 17, 2015, coll. J.H. Mirza; five males and four females paratypes, Hilaria spp. (Poaceae), Sanabal Farm, Kharaj, Riyadh, SA, 24\u00b016.999'N, 47\u00b011.854'E, January, 23, 2015, coll. M. Kamran.Holotype female, one male and two female paratypes, Remarks. The Paraplonobia (Anaplonobia) haloxylonia sp. n. closely resembles Paraplonobia (Anaplonobia) contiguus (Paraplonobia (Anaplonobia) contiguus by comparative length of leg I (shorter than body vs. longer than body), dorsocentral setae almost 1/3 to the distance of setae next behind vs. more than half, number of setae on genu I (5 vs. 4) in Paraplonobia (Anaplonobia) contiguus.ontiguus because Taxon classificationAnimaliaProstigmataTetranychidaehttp://zoobank.org/57BF2D3A-80B0-4C7E-90CD-FACB4543B5FFDorsal setae slightly lanceolate, densely serrate, not present on tubercles and distinctly shorter to the distances of setae next behind, prodorsum entirely with longitudinal striaitons, hysterosomal striations closely spaced, peritremes complex anastomosed distally, stylophore slightly rounded anteriorly, leg I shorter than body length, number of setae on femur I\u2013IV 8\u20136\u20133\u20133, number of setae on genu I\u2013IV 4\u20135\u20133\u20133.(n = 3). Measurements of holotype followed by 2 paratypes (in parenthesis) Figs \u201336.Dorsum , sc1 29 (28\u201331), sc2 30 (28\u201332), c1 28 (26\u201330), c2 26 (24\u201328), c3 29 (28\u201332), d1 23 (21\u201325), d2 22 (21\u201324), e1 21 (20\u201323), e2 22 (21\u201324), f1 23 (21\u201324), f2 26 (24\u201327), h1 27 (25\u201329). Distance between dorsal setae: v2\u2013v2 89 (85\u201390), v2\u2013sc1 68 (65\u2013690), sc1\u2013sc2 68 (67\u201370), sc1\u2013sc1 204 (202\u2013206), sc2\u2013sc2 301 298\u2013302), c1\u2013c1 138 (135\u2013140), c1\u2013c2 91 (89\u201392), c2\u2013c3 79 (75\u201380), c2\u2013c2 327 (325\u2013328), c3\u2013c3 424 (422\u2013426), d1\u2013d1 119 (118\u2013120), d1\u2013d2 91 (89\u201392), d2\u2013d2 295 (292\u2013298), c1\u2013d1 88 (86\u201389), c3\u2013d2 110 (109\u2013112), e1\u2013e1 27 (25\u201328), e1\u2013e2 85 (84\u201386), e2\u2013d2 85 (84\u201386), e2\u2013e2 229 (228\u2013231), f1\u2013f1 78 (76\u201380), f2\u2013f2 113 (110\u2013114), f1\u2013f2 35 (33\u201336), f1\u2013d1 82 (81\u201384), h1\u2013h1 53 (52\u201356).sum Fig. . Body roVenter , g2 39 (35\u201340), g1\u2013g1 52 (50\u201355), g2\u2013g2 60 (58\u201364), g1\u2013g2 12 (10\u201313); anal setae three pairs, ps1 20 (18\u201321), ps2 26 (24\u201327), ps3 28 (27\u201329), ps1\u2013ps1 23 (20\u201324), ps2\u2013ps2 32 (31\u201335), ps3\u2013ps3 23 (21\u201326); para-anal setae two pairs, h2 27 (26\u201328), h2\u2013 h2 11 (10\u201313), h3 32 (31\u201334), h3\u2013h3 28 (27\u201329).ter Fig. . IdiosomGnathosoma , 30 km Tabuk road, Sharma, Tabuk region, SA, 28\u00b003.479'N, 035\u00b017.186'E, October, 19, 2015, coll. M. Kamran and J.H. Mirza.Holotype female, two paratype females, Paraplonobia (Anaplonobia) tabukensis sp. n. closely resembles Paraplonobia (Anaplonobia) theroni (Paraplonobia (Anaplonobia) theroni by shape of stylophore anteriorly (rounded vs. slightly indented), number of setae on femur I\u2013IV (8\u20136\u20133\u20133 vs. 9\u20136\u20134\u20134), number of setae on genu I\u2013IV (4\u20135\u20133\u20133 vs. 5\u20135\u20136\u20136), number of setae on tibia III (8 vs. 6) and on tarsi I\u2013II excluding duplex setae and solenidia (10\u20137 vs. 18\u201314) in Paraplonobia (Anaplonobia) theroni.The theroni because theroni , 1987. TPageBreakTaxon classificationAnimaliaProstigmataTetranychidaeGenusWainstein, 1956Monoceronychus : Neopetrobia : Wainstein 1956: 151, Neopetrobiadubinini Wainstein, 1956.Based on Baker and Tuttle 1968, Gutierrez 1955, 1) widely spaced, not normal as c1; peritremes anastomosing distally.True claws pad like, each bearing a pair of tenant hairs; empodial pad longer than true claws, bearing a row of tenant hairs, distally not coalescent; dorsum with 3 pairs of prodorsal setae which are short and spindle shaped or spatulate; setal tubercles small or nonexistent; fourth pair of dorsocentral setae Monoceronychusmcgregori Pritchard & Baker, 1955.Neopetrobiamcgregori (Pritchard & Baker) Meyer, 1987. Female (n=9). Body oval; length of idiosoma 369\u2013372, maximum width 238\u2013241, length of body (gnathosoma + idiosoma) 430\u2013433.Dorsum . Length of intercoxal and coxal setae: 1a 18\u201319, 3a 19\u201320, 4a 15\u201316, 1b 30\u201331, 1c 13\u201314, 2b 16\u201317, 2c 10\u201313, 3b 15\u201317, 4b 11\u201312; aggenital setae (ag) 26\u201327, ag\u2013ag 38\u201339; genital setae two pairs, g1 17\u201318, g2 21\u201322, g1\u2013g1 41\u201342, g2\u2013g2 76\u201378, g1\u2013g2 21\u201322; anal setae three pairs, ps1 11\u201312, ps2 10\u201311, ps3 12\u201313, ps1\u2013ps1 11\u201313, ps2\u2013ps2 16\u201318, ps3\u2013ps3 11\u201313; para-anal setae two pairs, h2 11\u201313, h2\u2013 h2 7\u20139, h3 7\u20138, h3\u2013h3 17\u201319.ter Fig. . IdiosomGnathosoma , near exit10, King Abdullah Road, Riyadh, SA, 24\u00b045.826'N, 46\u00b045.470'E, September 07, 2015, coll. M. Kamran and E. M. Khan.12 females, Neopetrobiamcgregori was originally described very briefly under the genus Monoceronychus and has been only reported from Miami shores of Florida, USA . WorldwiPageBreak"} +{"text": "II complex with a -N-phenyl-carbamohydrazino\u00adthio\u00adate ligand is reported. The crystal structure shows DMSO mol\u00adecules bridging the complex units, building an one-dimensional H-bonded polymer.The synthesis and crystal structure of a Zn N-phenyl-2-hy\u00addra\u00adzine\u00adcar\u00adbo\u00adthio\u00adamide ligand with zinc acetate dihydrate in a 2:1 molar ratio yielded a yellow solid, which was crystallized from DMSO to obtain the title compound, [Zn(C17H16N3S)2]\u00b7C2H6OS. The ZnII ion is four-coordinated in a distorted tetra\u00adhedral environment by two deprotonated ligands. Each ligand acts as an N,S-donor, forming a five-membered metallacycle. The maximum deviation from the mean plane of the N\u2013N\u2013C\u2013S chelate group is 0.0029\u2005(14)\u2005\u00c5 for the N-donor atom of one ligand and 0.0044\u2005(14)\u2005\u00c5 for the non-coordinating N atom of the second. The dihedral angle between the planes of the two chelate groups is 72.80\u2005(07)\u00b0. Bond lengths in the ligands are compared with those in the crystal structure of the free ligand. In the crystal, complex mol\u00adecules are connected by dimethyl sulfoxide solvate mol\u00adecules via N\u2014H\u22efO hydrogen-bonding inter\u00adactions, building a one-dimensional hydrogen-bonded polymer along the a-axis direction. The S atom and one C atom of the dimethyl sulfoxide solvate mol\u00adecules are disordered over two sets of sites with an occupancy ratio of 0.6:0.4.The reaction of the II complex with the N-phenyl-2-hy\u00addra\u00adzine\u00adcar\u00adbo\u00adthio\u00adamidate ligand. Thio\u00adsemicarbazone derivatives are N,S-donors with a wide range of coordination modes and a variety of applications in biological inorganic chemistry /N21\u2014N22 = 1.393\u2005(3)\u2005\u00c5, N2\u2014C11 = 1.303\u2005(3)/N22\u2014C31 = 1.304\u2005(3)\u2005\u00c5 and C11\u2014S1 = 1.755\u2005(2)/C31\u2014S21 = 1.749\u2005(2)\u2005\u00c5.The acidic hydrogen of the hydrazine fragment is lost by the reaction with the acetate anion. The negative charge of the deprotonated ligand is delocalized over the N\u2014N\u2014C\u2014S entity, as indicated by their inter\u00admediate bond lengths. The bond lengths in the ligand are also affected by the coordination with the metal atom, especially the C\u2014S bond length, which is consistent with increased single-bond character. In the crystal structure of the free ligand is 58.25\u2005(11)\u00b0. In the second ligand, the corresponding angle is 49.99\u2005(11)\u00b0 between the C25\u2013C30 and C32\u2013C37 rings. In addition, the aliphatic rings are also not planar. The maximum deviation from the mean plane for the C1\u2013C5/C10 ring is 0.355\u2005(3)\u2005\u00c5 for C3 and for the C21\u2013C25/C30 ring the maximum deviation is 0.359\u2005(3)\u2005\u00c5 for C23, with both of the aliphatic rings having an envelope conformationII complex mol\u00adecules and the DMSO solvent mol\u00adecules build a monomeric entity. The DMSO mol\u00adecule bridges two complex mol\u00adecules via inter\u00admolecular N\u2014H\u22efO hydrogen-bonding inter\u00adactions, building a one-dimensional hydrogen-bonded polymer along the a-axis direction hy\u00addra\u00adzine\u00adcar\u00adbo\u00adthio\u00adamide dissolved in THF (2\u2005mmol/40\u2005mL) with zinc acetate dihydrate dissolved in ethanol (1\u2005mmol/30\u2005mL) was refluxed for 4\u2005h under continuous stirring. An orange solid was obtained, filtered and washed with ethanol. Suitable crystals for X-ray diffraction were obtained in DMSO by slow evaporation of the solvent.Starting materials were commercially available and used without further purification. The ligand synthesis was adapted from a procedure reported previously = 1.2 Ueq(C) (1.5 for methyl H atoms) using a riding model with C\u2014H = 0.95\u2005\u00c5 for aromatic, C\u2014H = 0.99\u2005\u00c5 for methyl\u00adene, C\u2014H = 0.98\u2005\u00c5 for methyl and N\u2014H = 0.88\u2005\u00c5. In the DMSO solvate mol\u00adecule, the S atom and methyl\u00adene C atom C42 and attached H atoms are disordered and were refined using a split model with an occupancy ratio of 0.4:0.6.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901500612X/im2462sup1.cifCrystal structure: contains datablock(s) I, publication_text. DOI: 10.1107/S205698901500612X/im2462Isup2.hklStructure factors: contains datablock(s) I. DOI: 1056177CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "All four phenyl rings in the two independent mol\u00adecules are arranged in a propeller-like conformation. The two mol\u00adecules exhibit S,R- and R,S- chirality and are connected via C\u2014H\u22efO inter\u00admolecular inter\u00adactions.In the title compound, C 34H28O4, the cyclo\u00adhexa\u00addiene ring has a screw-boat conformation with a torsion angle between the double bonds being on average ca 15\u00b0 [15.2\u2005(3) and \u221215.3\u2005(3) in the two independent mol\u00adecules]. All four phenyl rings in both mol\u00adecules are arranged in a propeller-like conformation. The two mol\u00adecules exhibit S,R- and R,S- chirality, respectively, and are connected via C\u2014H\u22efO inter\u00admolecular inter\u00adactions. In turn, these weakly bound dimers form the mol\u00adecular crystal.In the title compound, C Addition reactions of tetra\u00adphenyl\u00adcyclo\u00adpenta\u00addienone, often abbreviated to \u2018tetra\u00adcyclone\u2019, were reviewed by Allen 1945, 1962 \u25b8.This study provides an opportunity to investigate the geometry of 1,3-cyclo\u00adhexa\u00addiene rings surrounded by bulky substituents with no strong inter\u00admolecular inter\u00adactions.s-gauche \u2005\u00c5, \u03b8 = 115.8\u2005(3)\u00b0 and \u03c6 = 213.1\u2005(3); (C101\u2013C106) Q = 0.463\u2005(2)\u2005\u00c5, \u03b8 = 63.7\u2005(2)\u00b0 and \u03c6 = 33.5\u2005(3)\u00b0.The cyclo\u00adhexa\u00addiene rings and \u221215.3\u2005(3) for the two independent mol\u00adecules form short contacts with di\u00admethyl\u00admaleate following Allen & Sheps 1934. GC\u2013MS aUiso = 1.2iUso(C). All aromatic hydrogen atoms were refined with riding coordinates with C\u2014H = 0.95\u20130.98\u2005\u00c5 and Uiso = 1.2Uiso(C). Idealized methyl groups were refined as rotating groups with Uiso = 1.5Uiso(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989016009403/zl2665sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016009403/zl2665Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016009403/zl2665Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016009403/zl2665Isup4.cmlSupporting information file. DOI: 1484412CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds link complex mol\u00adecules, forming a three-dimensional network incorporating The title complex, [Mg(C 7H7N4O2)2(H2O)4], lies across an inversion centre and the MgII atom is coordinated in a slightly distorted octa\u00adhedral environment by four aqua ligands in the equatorial sites and two 1,3-dimethyl-2,6-dioxo-3,7-di\u00adhydro-1H-purin-9-ide ligands, through imidazole ring N atoms, in the axial sites. An intra\u00admolecular O\u2014H\u22efO hydrogen bond forms an S(7) graph-set motif. In the crystal, O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds link complex mol\u00adecules forming a three-dimensional network incorporating R42(8) and R22(18) graph-set motifs.The title complex, [Mg(C II ion in a basic solution to give rise to a tetra\u00adaqua mononuclear MgII complex, (I)Co-crystallization represents a crystal engineering approach for modifying properties of active pharmaceutical ingredients (APIs) Sun, 2013. Metal cII atom is coordin\u00adated in a slightly distorted octa\u00adhedral environment ], in the axial sites. The symmetry-unique purine ring system is essentially planar, with a maximum deviation of 0.030\u2005(2)\u2005\u00c5 for N3 and the bonded methyl C atoms C4 and C5 deviate from this mean plane by \u22120.118\u2005(3) and 0.136\u2005(2)\u2005\u00c5, respectively.The mol\u00adecular structure of (I)t Table\u00a01 by four et al., 1995iii = 2.829\u2005(3)\u2005\u00c5 and O4\u22efO1iv = 2.780\u2005(2)\u2005\u00c5; symmetry codes: (iii) \u2212x, \u2212y, \u2212z; (iv) x, y, z\u00a0+\u00a01] between a coordinating water mol\u00adecule and a carbonyl group of a symmetry-related theophylline group. The mononuclear units are connected into a layer parallel to (010) \u2005\u00c5; symmetry code: (ii) x, \u2212y\u00a0+\u00a0z\u00a0+\u00a0In the crystal, the coordinating water mol\u00adecules are involved in various hydrogen-bonding inter\u00adactions Table\u00a02. A (8) g0) Fig.\u00a02, which i0) Fig.\u00a02 by hydroA search of the Cambridge Structural Database in water (15\u2005ml) was then added. The resulting solution was kept in air and, after several days, colourless block-shaped crystals were obtained.Theophylline was dissolved in water (20\u2005ml). An aqueous solution (15\u2005ml) of NaOH was added slowly. MgClUiso(H) = 1.2Ueq(C) or 1.5Ueq(C). H atoms bonded to O atoms were located in difference Fourier maps and were refined with a distance restraint of O\u2014H = 0.87\u2005(1)\u2005\u00c5. The isotropic displacement parameters were refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015003758/lh5752sup1.cifCrystal structure: contains datablock(s) I, LOU. DOI: 10.1107/S2056989015003758/lh5752Isup2.hklStructure factors: contains datablock(s) I. DOI: 1050901CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Two \u03bc3-bridging I\u2212 anions are linked by three AgI ions, leading to the formation of a dicapped triangular motif with Ag\u22efAg separations in the range 3.0823\u2005(5)\u20133.2999\u2005(5)\u2005\u00c5. Each AgI atom exhibits a distorted tetra\u00adhedral geometry, with coordination to two I atoms and two P atoms from bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)methane ligands. In the crystal, the I\u2212 anion is linked to the ptu mol\u00adecule through two N\u2014H\u22efI hydrogen bonds [graph-set motif R21(6)]. These N\u2014H\u22efI hydrogen bonds, in addition to weak C\u2014H\u22efS and C\u2014H\u22efI hydrogen bonds, form zigzag chains along [010]. Two of the phenyl rings of two dppm ligands are disordered over two sets of sites with refined occupancies of 0.557\u2005(16) and 0.443\u2005(16).The title complex, [Ag For the al. 2015.3I2(C25H22P2)3]I\u00b7C7H8N2S = 0.039wR(F2) = 0.104S = 1.0814160 reflections983 parameters216 restraintsH-atom parameters constrainedmax = 1.16 e \u00c5\u22123\u0394\u03c1min = \u22121.04 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I. DOI: 10.1107/S2056989015017120/lh5785Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015017120/lh5785fig1.tif. DOI: The mol\u00adecular structure with displacement ellipsoids drawn at the 50% probability level. All H atoms and the minor component of disorder are omitted for clarity.Click here for additional data file.10.1107/S2056989015017120/lh5785fig2.tif. DOI: Part of the crystal structure showing inter\u00admolecular N\u2014H\u22efI, C\u2014H\u22efS and C\u2014H\u22efI hydrogen bonds as dashed lines, forming a chain along [010].1424053CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports6: Article number: 2086410.1038/srep20864; Published online: 02122016; Updated: 04292016This Article contains typographical errors.In the Results section under subheading \u2018Impact of ZLD1039 on breast cancer cell growth\u2019,50 values of 0.99\u2009\u00b1\u20090.23 and 0.089\u2009\u00b1\u20090.019\u2009\u03bcM, respectively\u201d.\u201cAmong the cell lines, MCF-7 and ZR-75-1 were the most sensitive to ZLD1039 with ICshould read:50 values of 0.99\u2009\u00b1\u20090.23 and 1.089\u2009\u00b1\u20090.019\u2009\u03bcM, respectively\u201d.\u201cAmong the cell lines, MCF-7 and ZR-75-1 were the most sensitive to ZLD1039 with ICIn Figure 3d, the x-axis \u20181\u20138 days of treatment\u2019 was incorrectly given as \u20180\u20137 days of treatment\u2019. The correct Figure 3d appears below as Fig.\u00a01"} +{"text": "Z\u2032 = 2) oriented almost perpendicular to each other [dihedral angle between the central core of each mol\u00adecule = 77.95\u2005(3)\u00b0]. The two mol\u00adecules exhibit similar conformations with an extended structure and are linked by bifurcated hydrogen bonds into a ribbon along the a-axis direction.The title compound crystallizes with the two mol\u00adecules in the asymmetric unit oriented almost perpendicular to each other [dihedral angle between the central core of each mol\u00adecule = 77.95\u2005(3)\u00b0]. The two mol\u00adecules exhibit similar conformations with an extended structure. An intra\u00admolecular C\u2014H\u22efN hydrogen bond occurs in each mol\u00adecule. The two mol\u00adecules are linked by a bifurcated N\u2014H\u22ef hydrogen bond involving the NH group in mol\u00adecule A as donor. They are further linked into a ribbon along the a-axis direction by further bifurcated N\u2014H\u22ef hydrogen bonds involving the NH group in mol\u00adecule B as donor. C\u2014H\u22efO inter\u00adactions are also observed.The title compound, C E)-(meth\u00adoxy\u00adimino){2-[(2-methyl\u00adphen\u00adoxy)meth\u00adyl]phen\u00adyl}acetate] derivatives are broad spectrum fungicides labeled A and B and shown in Fig.\u00a01A, the nitro group is coplanar with the p-nitro\u00adphenyl ring . The central ethane hydrazide moiety (N2A/N3A/C8A/O3A) is strictly planar with an r.m.s. deviation of 0.000\u2005\u00c5 for the fitted atoms. The dihedral angles between this moiety and the adjacent aromatic are 18.99\u2005(4)\u00b0 for the nitro\u00adbenzylidene ring (C1A\u2013C6A) and 62.20\u2005(4)\u00b0 for the benzene ring (C11A\u2013C16A).The title compound crystallizes with two mol\u00adecules in the asymmetric unit , 0.043\u2005(2) and 0.127\u2005(2)\u2005\u00c5, respectively]. The central ethane hydrazide moiety (N2B/N3B/C8B/O3B) is planar (r.m.s. deviation = 0.002\u2005\u00c5). The dihedral angles between this moiety and the adjacent aromatic rings are 12.43\u2005(4)\u00b0 for the nitrobenzylidene ring (C1B\u2013C6B) and 57.99\u2005(4)\u00b0 for the benzene ring (C11B\u2013C16B).In mol\u00adecule A and B are oriented almost perpendicular to each other, the dihedral angle between their central cores (atoms C7 N2 N3 and C8) being 77.95\u2005(3)\u00b0.Mol\u00adecules For both mol\u00adecules, bond lengths and angles are all within the normal ranges; however, comparisons with similar mol\u00adecules cannot be made as there are no similar overall structures although, of course, their fragments exist.A\u2014H17B\u22efN4A and C17B\u2014H17C\u22efN4B; Table\u00a01An intra\u00admolecular hydrogen bond . The mol\u00adecules are further linked into a ribbon along the a-axis direction a bifurcated N3B\u2014H3B\u22ef hydrogen bond involving the corresponding NH group in the other independent mol\u00adecule, as shown in Fig.\u00a02The two independent mol\u00adecules are linked by a bifurcated hydrogen bond Table\u00a01 between A search of the Cambridge Structural Database -2-Meth\u00adoxy\u00adimino-2-{2-[(2-methyl\u00adphen\u00adoxy)meth\u00adyl]phenyl}ethane\u00adhydrazide was refluxed with p-nitro\u00adbenzaldehyde in the presence of 5 drops of glacial acetic acid in 20\u2005ml absolute ethanol for about 10\u2005h to get a white-colored product. This was dissolved in methanol and white crystals were obtained by slow evaporation.(2isoU(H) = 1.5Ueq(C) for methyl H atoms and = 1.2eqU(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015004569/hg5434sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015004569/hg5434Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015004569/hg5434Isup3.cmlSupporting information file. DOI: 1052231CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The cations also form base pairs via N\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds.The N 12H10N5O+\u00b7NO3\u2212, the adenine unit has an N9-protonated N(7)\u2014H tautomeric form with non-protonated N1 and N3 atoms. The dihedral angle between the adenine ring system and the phenyl ring is 51.10\u2005(10)\u00b0. The typical intra\u00admolecular N7\u2014H\u22efO hydrogen bond with an S(7) graph-set motif is also present. The benzoyl\u00adadeninium cations also form base pairs through N\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds involving the Watson\u2013Crick face of the adenine ring and the C and O atoms of the benzoyl ring of an adjacent cation, forming a supra\u00admolecular ribbon with R22(9) rings. Benzoyl\u00adadeninum cations are also bridged by one of the oxygen atoms of the nitrate anion, which acts as a double acceptor, forming a pair of N\u2014H\u22efO hydrogen bonds to generate a second ribbon motif. These ribbons together with \u03c0\u2013\u03c0 stacking inter\u00adactions between the phenyl ring and the five- and six-membered adenine rings of adjacent mol\u00adecules generate a three-dimensional supra\u00admolecular architecture.In the title molecular salt, C N6-substituted adenine compounds continue to attract inter\u00adest due to their biological activity as they can act as plant hormones and have anti-allergenic, anti\u00adbacterial, anti\u00adviral and anti\u00adfungal properties et al., 1997et al., 2003et al., 2005et al., 2011Non-covalent inter\u00adactions, such as hydrogen bonding, halogen bonding and \u03c0\u2013\u03c0 inter\u00adactions play major roles in mol\u00adecular recognition and pharmaceutical drug design processes \u2014H tautomeric form with N9 protonated and N1, N3 non-proton\u00adated. The inter\u00adnal angles at N7 [C8\u2014N7\u2014C5 = 108.9\u2005(2)\u00b0] and N9 [C8\u2014N9\u2014C4 = 107.9\u2005(2)\u00b0] are similar as both carry hydrogen atoms ring motif. A similar bond was found in the crystal structure of the neutral N6-benzoyl adenine via N\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds \u2005\u00c5 . The other face of the phenyl ring makes offset \u03c0\u2013\u03c0 contacts with both the imidazole [Cg1\u22efCg3ii = 3.7213\u2005(17)\u2005\u00c5] and the pyrimidine rings , Fig.\u00a03Cg1, Cg2 and Cg3 are the centroids of the imidazole, pyrimidine and phenyl rings, respectively. Similar contacts are found in related structures s Table\u00a01 involvinN6-substituted adenines, adeninium salts and their metal complexes have been investigated in a variety of crystalline environments. Neutral mol\u00adecules include N6-benzyl\u00adadenine and N6-benzoyl\u00adadenine\u2013dl-tartaric acid (1:1). In these, the benzoyl\u00adadenine mol\u00adecule has a conformation similar to that reported for the neutral benzoyl\u00adadenine crystal structure , a few drops of nitric acid were added. The resulting solution was warmed over a water bath for half an hour and then kept at room temperature for crystallization. After a week colourless prismatic crystals of (I)To a hot methanol solution of Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015024871/sj5489sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015024871/sj5489Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989015024871/sj5489Isup3.pdfSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015024871/sj5489Isup4.cmlSupporting information file. DOI: 1444600CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The free 2,4-D anions are linked to the complex units through duplex water\u2013carboxyl\u00adate O\u2014H\u22efO hydrogen bonds through the coordinating water mol\u00adecules. In the crystal, inter-unit O\u2014H\u22efO hydrogen-bonding inter\u00adactions involving coordinating water mol\u00adecules as well as the solvent water mol\u00adecule (occupancy 0.5) with carboxyl\u00adate O-atom acceptors, give a layered structure lying parallel to (001), in which \u03c0\u2013\u03c0 ligand\u2013cation inter\u00adactions [minimum ring centroid separation = 3.6405\u2005(17)\u2005\u00c5] and a short O\u2014H\u22efCl inter\u00adaction are also found.In the crystal structure of the title magnesium salt of the phen\u00adoxy herbicide acetic acid , [Mg(C These include discrete monomeric {[MgL2(H2O)4] [L = 2-(2-fluoro\u00adphen\u00adoxy)acetate 5]\u00b7L }n (H2O)5](C8H5Cl2O3)\u00b70.5H2O, was obtained from the reaction of 2,4-D with MgCO3 in aqueous ethanol and its crystal structure is reported herein.The phen\u00adoxy\u00adacetic acids comprise an important group of chemicals which has among its members those ring-substituted representatives having selective herbicidal activity, 6 complex units have, as expected, essentially octa\u00adhedral stereochemistry [Mg\u2014O bond length range = 2.031\u2005(2)\u20132.094\u2005(2)\u2005\u00c5], comprising a carboxyl\u00adate O-donor from a monodentate 2,4-D\u2212 ligand and five water mol\u00adecules. The free 2,4-D\u2212 counter-anion is linked to the complex unit through an unusual duplex water\u2013carboxyl\u00adate O\u2014H\u22efO hydrogen-bonding association involving the coordinating water mol\u00adecules O1W and O2W being 179.0\u2005(2) and 174.8\u2005(2)\u00b0 (ligand A), and 175.7\u2005(2) and 178.7\u2005(2)\u00b0 (anion B), respectively]. This contrasts with the parent acid 2,4-D \u2005\u00c5] is also observed [for symmetry codes (i) and (iii), see: Table\u00a01In the crystal of the title compound, inter-unit O\u2014H\u22efO hydrogen-bonding inter\u00adactions Table\u00a01 involvin3 to 15\u2005ml of a hot aqueous solution of acetic acid (0.1\u2005mmol) in ethanol\u2013water (1:10 v/v). After completion of the reaction, excess MgCO3 was removed by filtration and the solution was allowed to evaporate at room temperature, providing colourless prisms of the title compound from which a specimen was cleaved for the X-ray analysis.The title compound was synthesized by the addition of excess MgCOUiso(H) = 1.5Ueq(O). Other H atoms were included in the refinement at calculated positions (aromatic C\u2014H = 0.95\u2005\u00c5 or methyl\u00adene 0.99\u2005\u00c5), with Uiso(H) = 1.2Ueq(C), using a riding-model approximation. The site-occupancy factor for the water mol\u00adecule of solvation was determined as 0.502\u2005(4) and was subsequently fixed at 0.50.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814019357/wm5045sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S1600536814019357/wm5045Isup2.hklStructure factors: contains datablock(s) I. DOI: 1021287CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Within the complex cation, the CrIII atom is coordinated by six O atoms of six urea ligands, displaying a slightly distorted octa\u00adhedral coordination environment. The Cr\u2014O bond lengths involving the urea ligands are in the range 1.9534\u2005(13)\u20131.9776\u2005(12)\u2005\u00c5. The Cr2O72\u2212 anion has a nearly staggered conformation, with a bridging angle of 130.26\u2005(10)\u00b0. The individual components are arranged in rows extending parallel to [100]. The Br\u2212 anion links the complex cation, as well as the solvent water mol\u00adecule, through N\u2014H\u22efBr and O\u2014H\u22efBr hydrogen-bonding inter\u00adactions. The supra\u00admolecular architecture also includes N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonding between urea N\u2014H and water O\u2014H donor groups and the O atoms of the Cr2O72\u2212 anion as acceptor atoms, leading to a three-dimensional network structure.The title bromide salt, [Cr{CO(NH On the other hand, CrVI is toxic and recognized as a carcinogen to humans and wildlife. The dichromate ion is environmentally important due to its high toxicity in aqueous solution have been investigated. It was found that [Cr(urea)6]3+ is suitable to target these oxoanions Cl\u00b7H2O has been reported , b = 10.393\u2005(1), c = 17.794\u2005(3)\u2005\u00c5 and \u03b2 = 94. 86\u2005(2)\u00b0. Within our broader study of CrIII complexes as industrial materials Br\u00b7H2O, (I)Counter-ionic species in coordination compounds play important roles in chemistry, pharmacy, mol\u00adecular assembly, biology and catalysis, as well as contributing significantly to environmental pollution; however, their binding characteristics have not received much recognition Cl\u00b7H2O investigated previously In order to check if compound (I)III ion is coordinated by six urea ligands through oxygen atoms with CrA\u2014OA bond lengths ranging from 1.9534\u2005(13) to 1.9776\u2005(12)\u2005\u00c5, and with OA\u2014CrA\u2014OA bond angles in the range 85.10\u2005(5)\u201392.95\u2005(5)\u00b0. The CrA\u2014OA bond lengths involving the urea ligand are in good agreement with the value of 1.9630\u2005(17)\u2005\u00c5 for [Cr(urea)6](BF4)3 2(cyclam)]ClO4 (cyclam)]ClO4 (cyclam)](ClO4)2\u00b70.5H2O 2(cyclam)]NO2 (acac)] \u00b0, 173.94\u2005(5)\u00b0, and 175.89\u2005(5)\u00b0, respectively. The bond lengths within the urea ligand are in the ranges of 1.263\u2005(2)\u20131.276\u2005(2) and 1.316\u2005(2)\u20131.328\u2005(2)\u2005\u00c5 for C=O and C\u2014N bonds, respectively. The C=O bonds are slightly longer than that in free non-coordinating urea 2Cr2O7, the tetra\u00adhedral CrO4 groups are in an almost eclipsed conformation and 1.8011\u2005(17)\u2005\u00c5 are longer than the terminal CrB\u2014OB bonds that are in the range of 1.6014\u2005(16)\u20131.6299\u2005(14)\u2005\u00c5. The Cr1B\u2014O7B\u2014Cr2B bridging angle in the complex anion is 130.26\u2005(10)\u00b0. The OB\u2014CrB\u2014OB bond angles in the two tetra\u00adhedral CrO4 groups are between 105.21\u2005(8) and 110.98\u2005(10)\u00b0, indicating slight angular distortions.It is of inter\u00adest to compare the conformation of Cr6]3+ moiety in compound (I)2O72\u2212 anions due to its high positive charge and the large number of hydrogen-bond donor groups of its six urea ligands.It is confirmed that the Br3\u00b73H2O was used as the starting material and was prepared according to literature procedures , dissolved in 10\u2005mL of water, was added to this solution. The mixture was refluxed at 353\u2005K for 10\u2005min and then cooled to room temperature. Green crystals of (I)2)2}6](Cr2O7)Br\u00b7H2O: C, 9.92; H, 3.61; N, 23.14%; found: C, 10.32; H, 3.08; N, 23.38%.All chemicals were reagent-grade materials and used without further purification. Chromium(III) tribromide hexa\u00adhydrate was obtained from Aldrich Chemical Co. and used as supplied. [Cr(urea)Uiso(H) = 1.2Ueq(N). H atoms of the solvent water mol\u00adecule were found from difference maps and refined with Uiso(H) = 1.2Ueq(O) and restrained to O\u2014H = 0.84\u2005(1) and H\u22efH = 1.36\u2005(2)\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015019258/wm5225sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015019258/wm5225Isup2.hklStructure factors: contains datablock(s) I. DOI: 1430688CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked 33H26F2N2O2, both pyrrolidine rings of the pyrrolizidine moiety adopt an envelope conformation. The di\u00adfluoro\u00adphenyl group is oriented at an angle of 54.3\u2005(1)\u00b0 with respect to the oxindole moiety. The crystal packing features an N\u2014H\u22efO hydrogen bond, which forms an R22(8) motif, and a C\u2014H\u22efO inter\u00adaction, which generates a C(8) chain along [010]. In addition, this chain structure is stabilized by C\u2014H\u22ef\u03c0 inter\u00adactions. In one of the pyrrolidine rings, the methyl\u00adene group forming the flap of an envelope and the H atoms of the adjacent methyl\u00adene groups are disordered over two sets of sites, with site-occupancy factors of 0.571\u2005(4) and 0.429\u2005(4)In the title pyrrolizidine derivative, C H-indole-2,3-dione) has been exploited extensively as a key inter\u00admediate in organic multicomponent reactions due to its anti\u00adbacterial -1\u2032,2,2\u2032,3,5\u2032,6\u2032,7\u2032,7a\u2032-octa\u00adhydro-1H-indan-2-spiro-3\u2032-(3\u2032H- pyrrolizine)-1\u2032-spiro-3\u2032\u2032-1H-indoline-1,2\u2032\u2032,3-trione (IV) sp3 hybridization. The fluorine atoms, F1 and F2, deviate by 0.006\u2005(2) and \u22120.010\u2005(2)\u2005\u00c5, respectively, from the plane of the benzene ring (C1\u2013C6) to which they are attached. The oxindole group system is planar with maximum deviations from its plane for the carbonyl C30 [\u22120.048\u2005(2)\u2005\u00c5] and O2 atoms [\u22120.122\u2005(1)\u2005\u00c5]. The di\u00adfluoro\u00adphenyl group is oriented at an angle of 54.3\u2005(1)\u00b0 with respect to the oxindole moiety. The benzene rings C7\u2013C12 and C21\u2013C26 are oriented at a dihedral angle of 52.7\u2005(1)\u00b0. The dihedral angles subtended by these two benzene rings with respect to the oxindole moiety are 21.2\u2005(1) and 31.6\u2005(1)\u00b0, respectively. The dihedral angle between the benzene rings of the biphenyl group is 44.3\u2005(1)\u00b0. Atom C18 of the pyrrolizidine ring system, and the adjacent methyl\u00adene group H atoms, are disordered over two sets of sites, with the site-occupancy factors of 0.571\u2005(4) and 0.429\u2005(4).The sum of the angles at N1 of the pyrrolizidine ring system (340\u00b0) is in accordance with q2 = 0.393\u2005(2)\u2005\u00c5 and \u03d5 = \u2212167.8\u2005(2)\u00b0 for N1/C20/C14\u2013C16 ring, and q2 = 0.280\u2005(3)\u2005\u00c5 and \u03d5 = 104.8\u2005(4)\u00b0 for N1/C16\u2013C19. In the N1/C20/C14\u2013C16 ring, atom C14 deviates by 0.594\u2005(2)\u2005\u00c5 from the least-squares plane through the remaining four atoms, whereas in the N1/C16-C19 ring, atoms C18 and C18\u2032 deviate by \u22120.401\u2005(5) and 0.434\u2005(4)\u2005\u00c5, respectively, from the plane through the remaining four atoms.In the pyrrolizidine ring system, both pyrrolidine rings adopt envelope conformations; the puckering parameters are: via N\u2014H\u22efO hydrogen bonds into inversion dimers, generating an C(8) chains propagating along [010]; see Fig.\u00a04C(8) chains propagating along [010]; see Fig.\u00a05The geometry of inter\u00adactions observed in this structure are given in Table\u00a01To a solution of isatin (1\u2005mmol) and L-proline (1\u2005mmol) in methanol (25\u2005ml), 1-[4-phen\u00adyl]3-phenyl\u00adprop-2-en-1-one (1\u2005mmol) was added and the solution was refluxed for 6\u20138\u2005h. The completion of reaction was monitored by thin layer chromatography. After completion, the reaction mixture was poured onto crushed ice. The precipitate obtained was filtered and dried at room temperature. Suitable crystals were obtained by slow evaporation of a solution of the title compound in aceto\u00adnitrile at room temperature.Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms. Atom C18 is disordered over two positions, with the major component having 0.571\u2005(4) occupancy. Pairs of C\u2014C distances were restrained to 1.54\u2005(1)\u2005\u00c5. The temperature factor of C18\u2032 was set to that of C18 with the EADP instruction of SHELXL2014/7 I, global. DOI: 10.1107/S2056989015012931/gk2637Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015012931/gk2637Isup3.cmlSupporting information file. DOI: 1410535CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The platinum(II) complex with notable antitumor activity shows a slightly distorted square-planar coordination and intramolecular C\u2014H\u22efCl and intermolecular N\u2014H\u22efCl and C\u2014H\u22efO hydrogen bonds. 10H6NO2)Cl(C5H11N)], crystallizes with one mol\u00adecule in the asymmetric unit. The PtII cation has a slightly distorted square-planar coordination environment defined by a chloride anion, the quinoline N atom and a carboxyl\u00adate O atom of the bidentate quinaldate ligand and a piperidine N atom. An intra\u00admolecular C\u2014H\u22efCl hydrogen bond occurs. In the crystal, mol\u00adecules are stacked into columns along the c axis by the formation of N\u2014H\u22efCl and C\u2014H\u22efO hydrogen bonds.The title compound, [Pt(C The Cl\u2212 and the PtII atoms are displaced from the least-squares plane of the quinoline ring and all other coord\u00adinating atoms by 0.2936\u2005(7) and 0.0052\u2005(1)\u2005\u00c5, respectively. The piperidine ring adopts a chair conformation and is almost perpendicular to the coordination plane of the PtII cation [dihedral angle between the best plane through the piperidine ring and the four atoms coordinating to the PtII cation = 79.66\u2005(13)\u00b0]. Bond lengths are normal and agree well with related platinum compounds ] , prepared according to the synthetic protocol of Da et al. (2001v/v) was added gradually while stirring at room temperature for 1\u2005h. The reaction mixture was stirred further for 4\u2005h. The precipitated yellow substance was filtered off and washed consecutively with a 0.1 M HCl solution (2 \u00d7 2\u2005ml), warm water (3 \u00d7 2\u2005ml) and cold ethanol (2\u2005ml). The product was then dried in a vacuum at 323\u2005K for 4\u2005h. The yield was 80%. Single crystals suitable for X-ray diffraction analysis were obtained by slow evaporation from an ethanol\u2013water (1:1 v/v) solution at room temperature. Positive ESI\u2013MS: m/z 1973 [4M + Na]+, 1483 [3M + Na]+, 998 [2M + Na]+, 510 [M + Na]+, 977 [2M + H]+, 489 [M + H]+; IR (KBr) cm\u22121: 3192 (\u03bdNH); 3080, 2930, 2866 (\u03bdCH); 1678 (\u03bdC=O); 1592, 1459 (\u03bdC=C arom); 1334 (\u03bdC\u2014O); 1H NMR : 9.50 , 8.51 , 8.06 , 7.91\u20137.88 , 7.71 , 3.52 , 3.27 = 12.5\u2005Hz, C510HNH), 1.76\u20131.61 , 4.00 .The starting complex K[PtCl al. 2001, was dis2 and N\u2014H = 0.93\u2005\u00c5 for amino H atoms, with Uiso = 1.2Ueq.All H atoms were refined using a riding model, with C\u2014H = 0.95\u2005\u00c5 for aromatic, C\u2014H = 0.99\u2005\u00c5 for CH10.1107/S160053681401191X/wm0005sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S160053681401191X/wm0005Isup2.hklStructure factors: contains datablock(s) I. DOI: 1004305CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Cu\u2014Cl bond measures 2.2287\u2005(9)\u2005\u00c5, and the two Cu\u2014S bonds are significantly different from each other, with values of 2.2270\u2005(10) and 2.2662\u2005(10)\u2005\u00c5. Also, the S\u2014Cu\u2014Cl angles differ, with values of 113.80\u2005(4) and 124.42\u2005(4)\u00b0, while the S\u2014Cu\u2014S angle is 121.51\u2005(4)\u00b0. The two imidazole rings are almost parallel, making a dihedral angle of 2.1\u2005(2)\u00b0. In the crystal, the shortest C\u2014H\u22efCl interactions stabilize a three-dimensional network with molecules linked into centrosymmetric dimers that are stacked along the b-axis direction.The mol\u00adecular structure of the title compound, [CuCl(C DOI: 10.1107/S1600536814024404/zq2228Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814024404/zq2228fig1.tif. DOI: Mol\u00adecular structure of the title compound with anisotropic displacement parameters drawn at the 50% probability level.1032971CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound comprises a 2-imino\u00adpyridine ring fused with a cyclo\u00adoctane ring, which adopts a twist boat\u2013chair conformation. Inter\u00admolecular C\u2014H\u22efN inter\u00adactions form 25H24ClN3, comprises a 2-imino\u00adpyridine ring fused with a cyclo\u00adoctane ring, which adopts a twist boat\u2013chair conformation. In the crystal, C\u2014H\u22efN inter\u00adactions form R22(14) ring motifs and mol\u00adecules are further connected by weak C\u2014H\u22ef\u03c0 inter\u00adactions. The resulting supra\u00admolecular structure is a two-dimensional framework parallel to the ab plane.The title compound, C They have gained importance in the medicinal and pharmaceutical fields due to their broad spectrum of biological activity, including anti-inflammatory, analgesic, anti\u00admicrobial, anti\u00adconvulsant, anti\u00adtubercular \u00b0 shows the linearity of the cyano group, a feature systematically observed in carbo\u00adnitrile compounds. Nitrile atoms C38 and N2 are displaced from the mean plane of the pyridine ring by 0.0258\u2005(1) and 0.0363\u2005(1)\u2005\u00c5, respectively. The imino C1=N1 bond length is 1.286\u2005(2)\u2005\u00c5. The imino group is nearly coplanar with the pyridine ring, as indicated by the N1=C1\u2014N3\u2014C5 torsion angle of \u2212178.89\u2005(14)\u00b0. The chloro\u00adbenzene ring is attached to the pyridine ring with a C2=C3\u2014C31\u00a0C36 torsion angle of 100.99\u2005(19)\u00b0, indicating a (+)anti\u00adclinal conformation. The C33\u00a0C34\u00a0C35 bond angle of 121.11\u2005(15)\u00b0 deviates from 120\u00b0 due to the presence of the chlorine substituent. The chlorine atom bonded to C34 deviates by 0.0446\u2005(1)\u2005\u00c5 from the mean plane of the phenyl ring. The chlorine is attached to the benzene ring with a C32\u00a0C33\u00a0C34\u2014Cl1 torsion angle of 178.95\u2005(13)\u00b0. In the pyridine ring, the formal double bonds [C4=C5 = 1.375\u2005(2) and C2=C3 = 1.369\u2005(2)\u2005\u00c5] are longer than standard C=C bonds (1.34\u2005\u00c5), while the other bond lengths are slightly shorter than standard C\u2014C and C\u2014N bond lengths, evidencing that there is a homo-conjugation effect for this ring.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01et al., 1995ab plane.In the crystal, pairs of C\u2014H\u22efN inter\u00adactions form b]pyrid\u00adine-3-carbo\u00adnitrile -5,6,7,8,9,10-hexa\u00adhydro\u00adcyclo\u00adocta\u00ad[p-toluene\u00adsulfonic acid (0.5\u2005mmol) was added. The reaction mixture was refluxed for 2\u20133\u2005h. After completion of the reaction (followed by thin-layer chromatography), the mixture was poured into crushed ice and extracted with ethyl acetate. The excess of solvent was removed under reduced pressure and the residue was chromatographed using a petroleum ether/ethyl acetate mixture (97:3 v/v) as eluent, to afford the pure product. The product was recrystallized from ethyl acetate, affording colourless crystals .Cyclo\u00adocta\u00adnone (1\u2005mmol), 4-chloro\u00adbenzaldehyde (1\u2005mmol) and malono\u00adnitrile (1\u2005mmol) were mixed in ethanol (10\u2005ml), and 2). Imine atom H1 was found in a difference map and refined freely, with the N\u2014H distance restrained to 0.84\u2005(2)\u2005\u00c5. Isotropic displacement parameters for H atoms were calculated as Uiso(H) = 1.2Ueq(C) for CH and CH2 groups, while the Uiso factor for H1 was refined. Crystal data, data collection and structure refinement details are summarized in Table\u00a02.C-bound H atoms were placed in calculated positions and allowed to ride on their carrier atoms, with C\u2014H = 0.93 (aromatic CH) or 0.97\u2005\u00c5 global, I. DOI: 10.1107/S160053681401962X/bh2503Isup2.hklStructure factors: contains datablock(s) I. DOI: 1021949CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "We used Sca1154Q/2Q mice, an established knock-in mouse model of the polyglutamine disease spinocerebellar ataxia type 1 (SCA1), which replicates human SCA1 features including ataxia, cognitive impairment, and neuronal death. We found that Sca1154Q/2Q mice exhibited greater synaptic instability than controls, without synaptic loss, in the cerebral cortex, where obvious neuronal death is not observed, even before the onset of distinct symptoms. Interestingly, this abnormal synaptic instability was evident in Sca1154Q/2Q mice from the synaptic developmental stage, and persisted into adulthood. Expression of synaptic scaffolding proteins was also lower in Sca1154Q/2Q mice than controls before synaptic maturation. As symptoms progressed, synaptic loss became evident. These results indicate that aberrant synaptic instability, accompanied by decreased expression of scaffolding proteins during synaptic development, is a very early pathology that precedes distinct neurological symptoms and neuronal cell death in SCA1.Late-onset neurodegenerative diseases are characterized by neurological symptoms and progressive neuronal death. Accumulating evidence suggests that neuronal dysfunction, rather than neuronal death, causes the symptoms of neurodegenerative diseases. However, the mechanisms underlying the dysfunction that occurs prior to cell death remain unclear. To investigate the synaptic basis of this dysfunction, we employed Many late-onset neurodegenerative diseases, including Alzheimer\u2019s, Parkinson\u2019s, and prion and polyglutamine diseases, share common features, such as the aggregation of toxic proteins in neurons, and progressive neuronal cell death1ATXN1 geneSca1154Q/2Q knock-in mouse model, harbouring 154 CAG repeats within the endogenous ATXN1 locus, closely reproduces the features of human SCA16Sca1154Q/2Q mice is much higher than that in human patients, another knock-in SCA1 mouse model harbouring 78 CAG repeats, similar to the number in patients, displays only mild behavioural deficits late in lifeSca1154Q/2Q mice are suitable for studying symptom progression. Sca1154Q/2Q mice develop motor learning impairment before any obvious Purkinje cell death occurs or nuclear inclusions form in the cerebellumSca1154Q/2Q mice show nuclear inclusions in pyramidal neurons, and cognitive deficits are observed without evident neuronal lossMany neurodegenerative diseases are attributed to multiple factors, including genetic and environmental predispositions. On the other hand, spinocerebellar ataxia type 1 (SCA1), a polyglutamine disease, is a monogenic disorder caused by the expansion of an unstable CAG trinucleotide repeat tract encoding a polyglutamine stretch in the Sca1154Q/2Q knock-in mice to elucidate the synaptic basis of neuronal dysfunction. We analysed the dynamics, morphology, and density of dendritic protrusions, which are excitatory postsynaptic structures classified into mature \u2018spines\u2019 and immature \u2018filopodia\u2019. These features are strongly associated with synaptic developmentSca1154Q/2Q knock-in mice in vivo, maintaining contributions from peripheral tissues and non-neuronal cells expressing mutant ataxin-1, as well as neurons. To evaluate neuronal dysfunction while excluding the effects of neuronal death, we focused on the cerebral cortex and hippocampus, in which apparent neuronal death does not occur despite the presence of cognitive dysfunction in both Sca1154Q/2Q mice and human SCA1 patients5In the present study, we focused on SCA1 as a genetic model of neurodegenerative disease, and used Sca1154Q/2Q mice are observed by 5 weeks of age, and spatial and fear memory deficits by 8 weeks. Although nuclear inclusions of mutant ataxin-1 are observed by 6 weeks, there is no neuronal death in the limbic area during such early stages of the diseaseSca1154Q/2Q mice as a possible early SCA1 phenotype. We performed in vivo two-photon imaging in layer 1 dendrites of the primary somatosensory cortex in Sca1154Q/2Q and control Sca12Q/2Q mice, and analysed the morphology, formation, and elimination of dendritic protrusions over a 1\u2009h period under anaesthesia vs Sca1154Q/2Q (0.33\u2009\u00b1\u20090.05/\u03bcm), p\u2009=\u20090.4532; spines: Sca12Q/2Q (0.31\u2009\u00b1\u20090.02/\u03bcm) vs Sca1154Q/2Q (0.24\u2009\u00b1\u20090.03/\u03bcm), p\u2009=\u20090.0909; filopodia: Sca12Q/2Q (0.070\u2009\u00b1\u20090.005/\u03bcm) vs Sca1154Q/2Q (0.09\u2009\u00b1\u20090.02/\u03bcm), p\u2009=\u20090.4159; unpaired t-test]. There were also no significant differences between 4-week-old Sca12Q/2Q and Sca1154Q/2Q mice in the number of spines or filopodia as a percentage of the total protrusions (Sca12Q/2Q (81\u2009\u00b1\u20091%) vs Sca1154Q/2Q (76\u2009\u00b1\u20092%), p\u2009=\u20090.0747; filopodia: Sca12Q/2Q (19\u2009\u00b1\u20091%) vs Sca1154Q/2Q (24\u2009\u00b1\u20092%), p\u2009=\u20090.0747; unpaired t-test]. Next, we analysed the dynamics of the protrusions in order to estimate synaptic stability. Notably, we found that the rates of formation and elimination of spines over 1\u2009h were significantly higher in Sca1154Q/2Q mice than in Sca12Q/2Q mice (Sca12Q/2Q (3.6\u2009\u00b1\u20090.9%) vs Sca1154Q/2Q (9\u2009\u00b1\u20092%), p\u2009=\u20090.0101; elimination rate: Sca12Q/2Q (4.5\u2009\u00b1\u20090.8%) vs Sca1154Q/2Q (12\u2009\u00b1\u20092%), p\u2009=\u20090.0017; unpaired t-test]. The elimination rate of filopodia over 1\u2009h was also significantly higher in Sca1154Q/2Q mice, but the difference in their formation rate did not reach statistical significance (Sca12Q/2Q (27\u2009\u00b1\u20096%) vs Sca1154Q/2Q (35\u2009\u00b1\u20094%), p\u2009=\u20090.2711; elimination rate: Sca12Q/2Q (23\u2009\u00b1\u20094%) vs Sca1154Q/2Q (42\u2009\u00b1\u20097%), p\u2009=\u20090.0341; unpaired t-test]. Anaesthetics can increase the formation of filopodia within a 1\u2009h periodin vivo imaging over 48\u2009h, during which time the mice were allowed to recover from the anaesthesia after the first imaging session and were returned to their home cages until the next session. We confirmed that both formation and elimination rates of spines in Sca1154Q/2Q mice were also significantly higher than those in Sca12Q/2Q mice over a 48\u2009h period (Sca12Q/2Q (8\u2009\u00b1\u20091%) vs Sca1154Q/2Q (23\u2009\u00b1\u20093%), p\u2009<\u20090.0001; elimination rate: Sca12Q/2Q (10\u2009\u00b1\u20091%) vs Sca1154Q/2Q (28\u2009\u00b1\u20094%), p\u2009=\u20090.0003; unpaired t-test]. Neither the formation nor elimination rates of filopodia over 48\u2009h were significantly different between groups, probably owing to the very high rates measured over 48\u2009h even in control mice (Sca12Q/2Q (59\u2009\u00b1\u20099%) vs Sca1154Q/2Q (52\u2009\u00b1\u20099%), p\u2009=\u20090.5763; elimination rate: Sca12Q/2Q (70\u2009\u00b1\u200910%) vs Sca1154Q/2Q (72\u2009\u00b1\u20097%), p\u2009=\u20090.8919; unpaired t-test]. These data indicate that Sca1154Q/2Q mice show abnormal synaptic instability during synaptic development, before the onset of obvious symptoms, and that these altered dynamics are detectable by imaging under anaesthesia during a 1\u2009h period.Motor learning impairments in esthesia . Dendrits of age . The number of spines as a percentage of total protrusions was significantly lower in Sca1154Q/2Q mice than in Sca12Q/2Q mice at 6 weeks (Sca12Q/2Q (88\u2009\u00b1\u20092%) vs Sca1154Q/2Q (81\u2009\u00b1\u20092%), p\u2009=\u20090.0284; unpaired t-test], whereas that of filopodia was higher (Sca12Q/2Q (12\u2009\u00b1\u20092% vs Sca1154Q/2Q (19\u2009\u00b1\u20092%), p\u2009=\u20090.0284; unpaired t-test]. Spine dynamics were significantly greater in Sca1154Q/2Q mice than in Sca12Q/2Q mice at 6 weeks of age (Sca12Q/2Q (2.3\u2009\u00b1\u20090.5%) vs Sca1154Q/2Q (9\u2009\u00b1\u20092%), p\u2009=\u20090.0018; elimination rate: Sca12Q/2Q (2.1\u2009\u00b1\u20090.6%) vs Sca1154Q/2Q (13\u2009\u00b1\u20092%), p\u2009<\u20090.0001; unpaired t-test]. Regarding the filopodium dynamics of 6-week-old Sca1154Q/2Q mice, the elimination rate was significantly higher, but the formation rate was not different from that in Sca12Q/2Q mice (Sca12Q/2Q (18\u2009\u00b1\u20097%) vs Sca1154Q/2Q (36\u2009\u00b1\u20096%), p\u2009=\u20090.0785; elimination rate: Sca12Q/2Q (14\u2009\u00b1\u20093%) vs Sca1154Q/2Q (35\u2009\u00b1\u20097%), p\u2009=\u20090.0100; unpaired t-test]. In 8-week-old Sca1154Q/2Q mice, total protrusion and spine densities were significantly lower than those in age-matched Sca12Q/2Q mice, whereas filopodium density was not different between the two groups (Sca12Q/2Q (0.36\u2009\u00b1\u20090.02/\u03bcm) vs Sca1154Q/2Q (0.30\u2009\u00b1\u20090.02/\u03bcm), p\u2009=\u20090.0343; spines: Sca12Q/2Q (0.33\u2009\u00b1\u20090.02/\u03bcm) vs Sca1154Q/2Q (0.25\u2009\u00b1\u20090.01/\u03bcm), p\u2009=\u20090.0175; filopodia: Sca12Q/2Q (0.033\u2009\u00b1\u20090.004/\u03bcm) vs Sca1154Q/2Q (0.044\u2009\u00b1\u20090.006/\u03bcm), p\u2009=\u20090.1621; unpaired t-test]. The number of spines as a percentage of the total protrusions was significantly lower in 8-week-old Sca1154Q/2Q mice than in Sca12Q/2Q mice (Sca12Q/2Q (90\u2009\u00b1\u20091%) vs Sca1154Q/2Q (85\u2009\u00b1\u20092%), p\u2009=\u20090.0403; unpaired t-test], whereas that of filopodia was higher in 8-week-old Sca1154Q/2Q mice than in Sca12Q/2Q mice (Sca12Q/2Q (10\u2009\u00b1\u20091%) vs Sca1154Q/2Q (15\u2009\u00b1\u20092%), p\u2009=\u20090.0403; unpaired t-test]. Both formation and elimination rates of spines in 8-week-old Sca1154Q/2Q mice were significantly higher than those in Sca12Q/2Q mice (Sca12Q/2Q (2.7\u2009\u00b1\u20090.7%) vs Sca1154Q/2Q (13\u2009\u00b1\u20092%), p\u2009<\u20090.0001; elimination rate: Sca12Q/2Q (2.0\u2009\u00b1\u20090.8%) vs Sca1154Q/2Q (12\u2009\u00b1\u20094%), p\u2009=\u20090.0176; unpaired t-test]. The formation rate of filopodia in 8-week-old Sca1154Q/2Q mice was higher than that in Sca12Q/2Q, whereas the elimination rate was not significantly different (Sca12Q/2Q (5\u2009\u00b1\u20093%) vs Sca1154Q/2Q (25\u2009\u00b1\u20098%), p\u2009=\u20090.0134; elimination rate: Sca12Q/2Q (21\u2009\u00b1\u20098% vs Sca1154Q/2Q (42\u2009\u00b1\u200911%), p\u2009=\u20090.1166; unpaired t-test]. These data indicate that SCA1 mice have immature dendritic morphology and a loss of dendritic protrusions associated with persisting synaptic instability and the progression of SCA1 symptoms.We demonstrated that synaptic instability occurs in SCA1 mice before the onset of distinct symptoms at 4 weeks of age. Next, we evaluated the progression of synaptic pathology in SCA1 mice until 8 weeks of age, by which time dendritic spines have normally stabilised and the density of filopodia has reached a minimumice at 6 and 8 weice at 6 . In 6-wee groups , whereas at 12 weeks, dendritic protrusion density was significantly lower in Sca1154Q/2Q mice than in Sca12Q/2Q mice (Sca12Q/2Q (1.53\u2009\u00b1\u20090.06/\u03bcm) vs Sca1154Q/2Q (1.28\u2009\u00b1\u20090.04/\u03bcm), p\u2009=\u20090.0011; unpaired t-test]. An abnormal frequency distribution of dendritic protrusion width was observed in Sca1154Q/2Q mice, particularly at 12 weeks of age, when the distribution curve shifted to the left . The mean dendritic protrusion width in 12-week-old Sca1154Q/2Q mice was lower than that in Sca12Q/2Q mice . These results show that the dendritic protrusion width in Sca1154Q/2Q mice decreased as SCA1 symptoms developed. An abnormal frequency distribution of protrusion length in Sca1154Q/2Q mice was evident at 5 weeks of age, and the frequency distribution curve was shifted to the right . No difference was observed in the mean length of dendritic protrusions between Sca1154Q/2Q and Sca12Q/2Q mice at either age . These results indicate that the protrusion lengths in Sca1154Q/2Q mice differ little from those in Sca12Q/2Q mice from early life through to adulthood. In summary, SCA1 mice demonstrated progressive deficits in dendritic protrusions in the hippocampus from adult ages./2Q mice . We choss of age with EDTA-free complete protease inhibitors and PhosSTOP (Roche Applied Science). Immunoblotting was performed using the following antibodies: anti-Homer1b/c , anti-pan-Shank , anti-PSD95 , anti-mGluR5 , anti-NR2B , anti-GluR1 , anti-Bassoon , and anti-\u03b2-actin . Total protein (10\u2009\u03bcg/lane) was separated on sodium dodecyl sulphate-polyacrylamide gels, transferred to immunoblot polyvinylidene difluoride membranes , incubated with 3% BSA in TBST for 1\u2009h at room temperature, and incubated with each primary antibody overnight at 4\u2009\u00b0C. The membranes were washed in TBST and further incubated with anti-mouse/rabbit IgG-horseradish peroxidase conjugate . After washing in TBST, the membranes were developed with Super Signal West Dura or Femto extended duration substrate (Pierce) and analysed using a ChemiImager . Proteins were identified by band position relative to a molecular weight marker.t-test, one-way ANOVA followed by Tukey multiple-comparison test, two-way ANOVA followed by Bonferroni multiple-comparison test, or the Kolmogorov\u2013Smirnov test. All statistical analyses were performed with GraphPad Prism . All data are presented as the mean\u2009\u00b1\u2009SEM.To determine statistical significance, we used the Student How to cite this article: Hatanaka, Y. et al. Abnormalities in synaptic dynamics during development in a mouse model of spinocerebellar ataxia type 1. Sci. Rep.5, 16102; doi: 10.1038/srep16102 (2015)."} +{"text": "In the title compound, the ring conformations of the tricycles are in an envelope, a half-chair and a chair. In the crystal, inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions link the mol\u00adecules into a three-dimensional architecture. 36H44O10\u00b7C6H6, the dioxolane ring adopts an envelope conformation with the C atom bonded to the H atom as the flap, while the cyclo\u00adhexene and cyclo\u00adhexane rings are in half-chair and chair conformations, respectively. In the crystal, a pair of O\u2014H\u22efO hydrogen bonds with an R22(26) graph-set motif connect the benzoate mol\u00adecules into an inversion dimer. The dimers are linked by a weak C\u2014H\u22efO inter\u00adaction into a tape structure along [01-1]. The benzene mol\u00adecule links the tapes through C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions, forming a sheet parallel to (100).In the title compound, C The title compound has been obtained in our synthetic study of paclitaxel as a cyclization precursor to build the taxane skeleton = 0.165\u2005(2)\u2005\u00c5 and \u03d5(2) = 114.5\u2005(6)\u00b0. The flap atom C4 deviates from the mean plane of other atoms by 0.270\u2005(3)\u2005\u00c5. The cyclo\u00adhexene ring (C5\u2013C10), which is spiro-fused to the dioxolane ring, is in a half-chair conformation with puckering parameters of Q = 0.469\u2005(2)\u2005\u00c5, \u03b8 = 127.5\u2005(2)\u00b0, \u03d5(2) = 197.2\u2005(3)\u00b0, Q(2) = 0.372\u2005(2)\u2005\u00c5 and Q(3) = \u22120.285\u2005(2)\u2005\u00c5. Atoms C5 and C6 deviate from the mean plane of the other atoms by \u22120.493\u2005(4) and 0.212\u2005(4)\u2005\u00c5, respectively. The cyclo\u00adhexane ring (C24\u2013C29) is in a chair conformation with puckering parameters Q = 0.587\u2005(2)\u2005\u00c5, \u03b8 = 4.6\u2005(2)\u00b0, \u03d5 = 246\u2005(3)\u00b0, Q(2) = 0.042\u2005(2)\u2005\u00c5 and Q(3) = 0.585\u2005(2)\u2005\u00c5. The large substituents are in the equatorial positions. The meth\u00adoxy\u00admeth\u00adoxy group (O42/C43/O44/C45) shows a helical form with torsion angles of 76.5\u2005(3)\u00b0 for C29\u2014O42\u2014C43\u2014O44 and 64.8\u2005(3)\u00b0 for O42\u2014C43\u2014O44\u2014C45 held by weak intra\u00admolecular C\u2014H\u22efO inter\u00adactions was provided according to the reported procedure (Nicolaou Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C). The H atom of hy\u00addroxy group (O41) was placed guided by difference maps and then treated as riding, with O\u2014H = 0.84\u2005\u00c5 and with Uiso(H) = 1.5Ueq(O). 13 problematic reflections were omitted from the final refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989014026048/is5382sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989014026048/is5382Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989014026048/is5382Isup3.cmlSupporting information file. DOI: 1036428CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit contains two Co2+ ions, one L3\u2212 anion originating from 5-(4-carb\u00adoxy\u00adphen\u00adoxy)isophthalic acid (H3L), one OH\u2212 ligand, one 1,4-bis\u00ad[(1H-imidazol-l-yl)meth\u00adyl]benzene (bix) ligand and one disordered lattice water mol\u00adecule (occupancy 0.25). The two Co2+ ions have different environments. One has an octa\u00adhedral O4N2 coordin\u00adation sphere, defined by four O atoms from three carboxyl\u00adate groups and one OH\u2212 ligand, and two N atoms from two symmetry-related bix ligands. The other has a trigonal-bipyramidal O5 coordination sphere resulting from three carboxyl\u00adate groups and two OH\u2212 ligands. The dihedral angles between the two benzene rings in the L3\u2212 ligand and between the benzene ring and the two imidazole rings in the bix ligand are 67.05\u2005(15), 75.27\u2005(17) and 82.05\u2005(17)\u00b0, respectively. Four neighbouring Co2+ ions are linked by six carboxyl\u00adate groups and two \u03bc3-OH ligands, forming a butterfly-shaped secondary building unit (SBU). These SBUs are connected by L3\u2212 anions into layers parallel to (1-10). Adjacent layers are cross-linked by the bix ligands, forming a three-dimensional framework that has a bimodal -connected tfz-d topology. The disordered lattice water mol\u00adecule is located in the voids of the framework and has O\u22efO and O\u22efN contacts of 2.81\u2005(2) and 2.95\u2005(2)\u2005\u00c5, suggesting medium-strength hydrogen bonds. The title compound may be a good candidate for artificial eye lenses.The title coordination polymer, {[Co For top al. 2010.2(C15H7O7)(OH)(C14H14N4)]\u00b70.25H2O = 0.037wR(F2) = 0.086S = 1.045314 reflections396 parameters1 restraintH atoms treated by a mixture of independent and constrained refinementmax = 0.37 e \u00c5\u22123\u0394\u03c1min = \u22120.37 e \u00c5\u22123\u0394\u03c1APEX2 I, New_Global_Publ_Block. DOI: 10.1107/S1600536814022806/wm5040Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814022806/wm5040fig1.tifx y z x y z x y z x y z x y z . DOI: x, 2\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z; ii) 2\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z; iii) \u22121\u00a0+\u00a0x, \u22121\u00a0+\u00a0y, z; iv) 2\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y, \u2212z; v) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, z.]The extended asymmetric unit of the title compound. Displacement ellipsoids are drawn at the 50% probability level. The disordered lattice water mol\u00adecule has been omitted for clarity. The tetra\u00adnuclear SBU in the structure of the title compound. [Symmetry code: A) 1\u00a0\u2212\u00a0Click here for additional data file.10.1107/S1600536814022806/wm5040fig3.tifL 3\u2212 . DOI: L3\u2212 anions.View of the layered network formed by the SBUs and the 1029647CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The geometry of the title hemibi\u00adquinone is different from previous examples and may be correlated with the weak inter\u00adactions in the crystal. 16H9BrN2O4\u00b70.5C6H6, the mol\u00adecules stack in a centrosymmetric unit cell in a 2:1 stoichiometry with co-crystallized benzene solvent mol\u00adecules and inter\u00adact via various weak inter\u00adactions. This induces a geometry different from that predicted by theory, and is unique among the hemibi\u00adquinones heretofore reported.In the crystal of the title compound, C Biphenyl derivatives have garnered great attention to themselves as conductors of electricity mol\u00adecule, 4\u2032-bromo-2\u2032,5\u2032-dimeth\u00adoxy-2,5-dioxo--3,4-dicarbo\u00adnitrile 1 Fig.\u00a01, has beenter 1974, where aet al., 20152(CN)2], Fig.\u00a021 is comparable to that in the hydro\u00adquinone [1.481\u2005(2)\u2005\u00c5]. In this mol\u00adecule, the C5\u2014C4\u2014C7\u2014C8 torsion is 125.3\u2005(2)\u00b0, compared to the hydro\u00adquinone torsion angle of \u2212126.50\u00b0, \u00b0 and C5\u2014C6\u2014O1\u2014C14 bent out of plane by \u22127.0\u2005(3)\u00b0. The meth\u00adoxy group bond angle C3\u2014O2\u2014C13 is measured at 117.4\u2005(1)\u00b0, and C6\u2014O1\u2014C14 is measured at 117.3\u2005(1)\u00b0. The methyl portions of each of these groups point away from the sterically restricting groups 1 [1.346\u2005(2)\u2005\u00c5] is shorter than than the corresponding hydro\u00adquinone C9\u2014C10 bond [1.408\u2005(2)\u2005\u00c5] and the C1\u2014C6 bond [1.334\u2005(6)\u2005\u00c5] of BrHBQBr \u2005\u00c5] compared to the same bond in the hydro\u00adquinone precursor [1.885\u2005(1)\u2005\u00c5], but is still shorter than that of the starting material BrHBQBr . Additionally, there is a short contact between C10 and C8, at 3.486\u2005(3)\u2005\u00c5. The N2 nitrile atom possibly accepts a very weak inter\u00adaction from the meth\u00adoxy C13 and H13A pair (2.82\u2005\u00c5 and 97\u00b0). There is a long C13\u22efN2 [(3.101\u2005(3)\u2005\u00c5] inter\u00adaction as well. Even longer than those inter\u00adactions, H13 also has a weak H\u22ef\u03c0 inter\u00adaction with the di\u00admeth\u00adoxy\u00adbenzene ring on an adjacent mol\u00adecule (H\u22ef\u03c0 = 2.88\u2005\u00c5). The packing is shown in Fig.\u00a05Each HBQ mol\u00adecule inter\u00adacts with a total of three benzene mol\u00adecules by short contacts. As mentioned above, one mol\u00adecule of benzene is sandwiched between two quinone rings. Additionally, the 3-substituted nitrile group accepts a C\u2014H\u22efN hydrogen bond from a solvent mol\u00adecule . The third benzene mol\u00adecule exhibits short contacts to the 4\u2032-bromine atom on the opposite end of the mol\u00adecule, where H17 and H18 link to Br1 almost symmetrically . Since the benzene mol\u00adecule \u03c0-stacks parallel to the quinones, the benzene mol\u00adecule is oriented in the same direction relative to the di\u00admeth\u00adoxy\u00adbenzene ring. Although, in previous HBQ crystals the 4 and 4\u2032 groups show evidence of inter\u00admolecular halogen bonding, due to the excess electron density around the aryl bromine atoms and the nitrile groups, an attractive inter\u00adaction is not possible, rather a slightly repulsive inter\u00adaction is favored. Instead, the protons on C17 and C18 bifurcate to Br1 as an acceptor, forming slightly asymmetric hydrogen bonds between the di\u00admeth\u00adoxy\u00adbenzene ring and the benzene solvent mol\u00adecule. As discussed above, the quinone carbonyl groups are deflected from perfect planarity. In previous structures, meth\u00adoxy oxygen atoms tended to deflect the carbonyl groups through repulsive effects. However, this structure contains some attractive inter\u00admolecular hydrogen bonding character, including the C14\u2014H142O and 100\u2005mL of benzene. FeCl3 was added in one portion. The resulting mixture was capped and stirred overnight. The resulting phases were separated, and the organic phase was washed with water and dried over anhydrous Na2SO4. Evaporation of the solvent produced a crude product. The pure product was precipitated from a chloro\u00adform solution by addition of hexane, yielding 0.0460\u2005g (36.7%). Black, block-shaped crystals of 1 were grown from chloro\u00adform solution with residual benzene at 296\u2005K. 1H NMR \u03b4 = 7.22 , 7.12 , 6.71 , 3.87 , 3.76 .4\u2032-Bromo-2,5-dihy\u00addroxy-2\u2032,5\u2032-dimeth\u00adoxy--3,4-dicarbo\u00adnitrile was suspended in a mixture of 100\u2005mL of H3, 0.88\u2005\u00c5. Methyl H atoms were allowed to rotate but not to tip to best fit the experimental electron density. Uiso(H) values were set to a multiple of Ueq(C) with 1.5 for CH3. Crystal data, data collection and structure refinement details are summarized in Table\u00a01H atoms attached to carbon atoms were positioned geometrically and constrained to ride on their parent atoms, with C\u2014H-bond distances of 0.95\u2005\u00c5 for aromatic C\u2014H, 1.00, 0.99 and 0.98\u2005\u00c5 for aliphatic CH10.1107/S2056989016005120/hb7567sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016005120/hb7567Isup2.hklStructure factors: contains datablock(s) I. DOI: 1470649CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two 2,2-di\u00adallyl\u00admalonate anions bridge four AgI ions in a \u03bc4- mode, setting up an Ag4O8P4 core (point group symmetry -4..) of corner-sharing tetra\u00adhedra. The shortest intra\u00admolecular Ag\u22efAg distance of 3.9510\u2005(3)\u2005\u00c5 reveals that no direct d10\u22efd10 inter\u00adactions are present. Four weak intra\u00admolecular C\u2014H\u22efO hydrogen bonds are observed in the crystal structure of the title compound, which most likely stabilize the tetra\u00adnuclear silver core.In the tetra\u00adnuclear mol\u00adecule of the title compound, [Ag On a mol\u00adecular level this is typically achieved by designing low aggregated metal compounds. In the case of silver, this can be realized by the use of phosphanes as a Lewis base; however, the concomitant increase of the mol\u00adecular weight of the transition metal complex may decrease its vapour pressure. Circumventing this difficulty, we have investigated the use of olefines as ligands for silver(I) carboxyl\u00adates, in which the olefin is covalently bonded to the carboxyl\u00adate. In the context of this approach, the title compound [{ mode \u2005\u00c5], which is supported by the respective bond angles around Ag1 (Table\u00a01The asymmetric unit of (I)de Fig.\u00a01. There i1 Table\u00a01 summing 1 Table\u00a01. However3P tetra\u00adhedra gives the tetra\u00adnuclear structure of (I)4O4 heterocubanes O,O\u2032)-, - or -chelation Ag}4{(O2C)2C(CH2CH=CH2)2}2] was prepared by the addition of PPh3 to a suspension of [(AgO2C)2C(CH2CH=CH2)2] in di\u00adchloro\u00admethane (30\u2005ml) at 273\u2005K. After stirring for 2\u2005h at this temperature, the reaction mixture was filtered through a pad of celite. Afterwards, all volatiles were removed in oil-pump vacuum, and (I)2C)2C(CH2CH=CH2)2]).Complex [{(Ph90H80Ag4O8P4 (1844.96): C 58.59, H 4.37. Found: C 58.53, H 4.34. 1H NMR : \u03b4 = 2.79 , 4.97 , 5.03 , 5.90 , 7.30\u20137.51 . 31P{1H} NMR : \u03b4 = 15.7 . IR : \u03bd = 1637 , 1559 , 1440 , 692 (vs), 521 (vs).Analysis calculated for CUiso(H) = 1.2Ueq(C) and a C\u2014H distance of 0.93\u2005\u00c5 for aromatic and vinylic as well as 0.97\u2005\u00c5 for methyl\u00adene protons. The unit cell contains two voids of 34(1.4)\u2005\u00c53. Void volume calculation using the SQUEEZE routine in PLATON ; however, this ambiguity is resolved as the Flack parameter of the inverted structure is calculated to 1.052\u2005(9). This indicates that the original absolute structure has been assigned correctly.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S1600536814019394/wm5047sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814019394/wm5047Isup2.hklStructure factors: contains datablock(s) I. DOI: 1021407CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The chloride ion occupies an equatorial site and the dihedral angle between the 2,2\u2032-bpy ring systems is 72.02\u2005(6)\u00b0. In the crystal, the components are linked by C\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions [shortest centroid\u2013centroid separation = 3.635\u2005(2)\u2005\u00c5], generating a three-dimensional network.In the title hydrated salt, [NiCl(C DOI: 10.1107/S1600536814009064/hb7220Isup2.hklStructure factors: contains datablock(s) I. DOI: 998760CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the structure of bis\u00ad(2-amino-5-chloro\u00adpyridinium) tetra\u00adchlorido\u00adcobaltate(II), the essentially planar cations are connected through N\u2014H\u22efCl hydrogen bonds to the tetra\u00adhedral anion. 5H6ClN2)2[CoCl4], was synthesized by slow evaporation of an aqueous solution at room temperature. The asymmetric unit consists of two essentially planar (C5H6ClN2)+ cations [maximum deviations = 0.010\u2005(3) and 0.014\u2005(3)\u2005\u00c5] that are nearly perpendicular to each other [dihedral angle = 84.12\u2005(7)\u00b0]. They are bonded through N\u2014H\u22efCl hydrogen bonds to distorted [CoCl4]2\u2212 tetra\u00adhedra, leading to the formation of undulating layers parallel to (100). The structure is isotypic with the Zn analogue The title salt, (C The bond angles C4\u2014N2\u2014C5 [123.6\u2005(3)\u00b0] and C9\u2014N3\u2014C10 [123.3\u2005(3)\u00b0] in the rings of cat1 and cat2, respectively, confirm the presence of pyridinium cations. Previous studies to 2.2934\u2005(12)\u2005\u00c5 and the Cl\u2014Co\u2014Cl angles range from 104.84\u2005(5) to 118.58\u2005(5)\u00b0, revealing considerable distortions from the ideal tetra\u00adhedral geometry. These values are in agreement with those observed in similar compounds by hydrogen bonds with a second symmetry-related cat1 cation. The hydrogen-bonding environments of the two cations are similar. Both are linked to two CoCl4 tetra\u00adhedra by three hydrogen bonds 2[ZnCl4]\u00b7H2O 2[ZnCl4] 2[CdCl4]\u00b7H2O 2[CuCl4] 2[CuBr4] 2[ZnCl4] chloride and 2-amino-5-chloro\u00adpyridine (molar ratio 1:1) was dissolved in an aqueous solution of hydro\u00adchloric acid with 5\u2005ml of ethanol. The mixture was stirred and then kept at room temperature. Blue crystals of the title compound were obtained after two weeks.Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015007707/wm5146sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989015007707/wm5146Isup2.hklStructure factors: contains datablock(s) I. DOI: 990478CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The ReI ion is coordinated by two N atoms from the 1,10-phenanthroline ligand and one N atom from the 1,2-bis\u00ad(pyridin-4-yl)ethane ligand [mean Re\u2014N = 2.191\u2005(15)\u2005\u00c5] and by three carbonyl ligands [mean Re\u2014C = 1.926\u2005(3)\u2005\u00c5] in a distorted octa\u00adhedral geometry. The electrostatic forces and weak C\u2014H\u22efF(O) hydrogen bonds pack cations and anions into the crystal with voids of 82\u2005\u00c53, which are filled by solvent mol\u00adecules. The crystal packing exhibits short inter\u00admolecular O\u22efO distance of 2.795\u2005(5)\u2005\u00c5 between two cations related by inversion.The asymmetric unit of the title compound, [Re(C DOI: 10.1107/S1600536814014135/cv5465Isup2.hklStructure factors: contains datablock(s) I. DOI: 1008626CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The equatorial plane is formed by two N,O-bidentate 1H-pyrazole-3-carboxyl\u00adate ligands in a trans configuration. The axial positions are occupied by two water mol\u00adecules. The mononuclear complex mol\u00adecules are arranged in layers parallel to the ab plane. Each complex mol\u00adecule is linked to four adjacent species through inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds that are established between the coordinating water mol\u00adecules and carboxyl\u00adate O atoms or protonated N atoms of the organic ligands. These layers are further connected into a three-dimensional network by additional hydrogen bonds involving solvent water mol\u00adecules and non-coordinating carboxyl\u00adate O atoms.In the title compound, [Cu(C H-pyrazole-3-carboxyl\u00adate ligand, see: Artetxe et al. , nickel(II) and zinc complexes of the 1 al. 2015; L\u00f3pez-V al. 2014.4H3N2O2)2(H2O)2]\u00b72H2O = 0.028wR(F2) = 0.074S = 1.091216 reflections109 parameters4 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.36 e \u00c5\u22123\u0394\u03c1min = \u22120.35 e \u00c5\u22123\u0394\u03c1CrysAlis PRO used to solve structure: OLEX2 I, global. DOI: 10.1107/S2056989015021593/im2473Isup2.hklStructure factors: contains datablock(s) global, a2013072_cupyc. DOI: Click here for additional data file.10.1107/S2056989015021593/im2473fig1.tif4 3 2 2 2 2 2 2 . DOI: 4H3N2O2)2(H2O)2] \u00b72H2O showing the atom labelling for the asymmetric unit and 50% probability displacement ellipsoids.Mol\u00adecular structure of [Cu. Projection of a layer of [Cu(C4H3N2O2)2(H2O)2] complexes along the [010] direction (below). Cu(II) centres are represented as translucent octa\u00adhedra and the O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds are depicted as dashed red lines.View of the crystal packing along the crystallographic 1437048CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "X is said to be End-completely-regular if its endomorphism monoid End(X) is completely regular . In this paper, we will show that if X[Y] is End-completely-regular , then both X and Y are End-completely-regular . We give several approaches to construct new End-completely-regular graphs by means of the lexicographic products of two graphs with certain conditions. In particular, we determine the End-completely-regular and End-inverse lexicographic products of bipartite graphs.A graph Endomorphism monoids of graphs are generalizations of automorphism groups of graphs. In recent years, much attention has been paid to endomorphism monoids of graphs and many interesting results concerning graphs and their endomorphism monoids have been obtained. The aim of this research is to develop further relationship between graph theory and algebraic theory of semigroups and to apply the theory of semigroups to graph theory. The bipartite graph is a class of famous graphs. Their endomorphism monoids are studied by several authors. In , the conX considered in this paper are undirected finite simple graphs. The vertex set of X is denoted by V(X) and the edge set of X is denoted by E(X). If two vertices x1 and x2 are adjacent in X, the edge connecting x1 and x2 is denoted by {x1, x2} and we write {x1, x2} \u2208 E(X). A subgraph H is called an induced subgraph of X if for any a, b \u2208 V(H), {a, b} \u2208 E(H) if and only if {a, b} \u2208 E(X). A graph X is called bipartite if X has no odd cycle. It is known that if a graph X is a bipartite graph, then its vertex set can be partitioned into two disjoint nonempty subsets such that no edge joins two vertices in the same set.The graphs X and Y be two graphs. The join of X and Y, denoted by X + Y, is a graph such that V(X + Y) = V(X) \u222a V(Y) and E(X + Y) = E(X) \u222a E(Y)\u222a{{x1, x2} | x1 \u2208 V(X), x2 \u2208 V(Y)}. The lexicographic product of X and Y, denoted by X[Y], is a graph with vertex set V(X[Y]) = V(X) \u00d7 V(Y), and with edge set E(X[Y]) = {{, }\u2223{x, x1} \u2208 E(X), or x = x1 and {y, y1} \u2208 E(Y)}. Denote Yx = { | y \u2208 V(Y)} for any x \u2208 V(X).Let X and Y be graphs. A mapping f from V(X) to V(Y) is called a homomorphism (from X to Y) if {x1, x2} \u2208 E(X) implies that {f(x1), f(x2)} \u2208 E(Y). A homomorphism f is called an isomorphism if f is bijective and f\u22121 is a homomorphism. A homomorphism f from X to itself is called an endomorphism of X (see \u03c1f the equivalence class containing a \u2208 V(X) with respect to \u03c1f.Aa of a semigroup S is called regular if there exists x \u2208 S such that axa = a. An element a of a semigroup S is called completely regular if a = axa and xa = ax hold for some x \u2208 S. A semigroup S is called regular if all its elements are regular . An inverse semigroup is a regular semigroup in which the idempotents commute. A graph X is said to be End-regular if its endomorphism monoid End(X) is regular . Clearly, End-completely-regular graphs as well as End-inverse graphs are End-regular.An element For undefined notation and terminology in this paper, the reader is referred to \u201314. We lX[Y] is End-regular, then both X and Y are End-regular.If G be a graph and let f \u2208 End(G). Then f is completely regular if and only if f|If \u2208 Aut(If).Let X be a bipartite graph. Then X is End-completely-regular if and only if X is one of K1, K2, P2, 2K1, 2K2, and K1 \u22c3 K2.Let X and Y be two bipartite graphs. Then X + Y is End-completely-regular if and only if one of them is End-completely-regular and the other is K1 or K2.Let G be a graph and f \u2208 End(G). Then f is completely regular if and only if there exists g \u2208 Idpt(G) such that \u03c1g = \u03c1f and Ig = If.Let X be a bipartite graph. Then X is End-inverse if and only if X = K1 or X = K2.Let X and Y be two graphs. Then End(X[Y]) = End(X)[End(Y)] if and only if for any f \u2208 End(X[Y]) and x \u2208 V(X), there exists x\u2032 \u2208 V(X) such that f(Yx)\u2286Yx\u2032.Let X[Y] is End-completely-regular, then both X and Y are End-completely-regular.In this section, we will characterize the End-completely-regular and End-inverse lexicographic products of two graphs. We first show that if X and Y be two graphs. If X[Y] is End-completely-regular, then both X and Y are End-completely-regular.Let X is End-completely-regular, it is only necessary to verify that f|If is an automorphism of If for each f \u2208 End(X). Define a mapping F from V(X[Y]) to itself byF \u2208 End(X[Y]). Since X[Y] is End-completely-regular, by F|IF is an automorphism of IF. It is easy to see that IF = If[Y]. For any distinct x1, x2 \u2208 V(If) and y \u2208 V(Y), F) = (f(x1), y) and F) = (f(x2), y) hold. Since F|IF is an automorphism of IF, (f(x1), y)\u2260(f(x2), y). Hence f(x1) \u2260 f(x2) and so f|If is an automorphism of If.By g \u2208 End(Y). Define a mapping G from V(X[Y]) to itself byG \u2208 End(X[Y]). Since X[Y] is End-completely-regular, by G|IG is an automorphism of IG. It is easy to see that IG = X[Ig]. For any x \u2208 V(X) and y1, y2 \u2208 V(Ig), G) = ) and G) = ). Since G|IG is an automorphism of IG, )\u2260), we get that g(y1) \u2260 g(y2) and so g|Ig is an automorphism of Ig, as required.Let X and Y being End-completely-regular does not yield that X[Y] is End-completely-regular.The following example shows that X and Y be two graphs with V(X) = {x1, x2}, V(Y) = {y1, y2}, E(X) = {{x1, x2}}, and E(Y) = \u03d5. By X and Y are End-completely-regular. It is easy to see that X[Y]\u2245C4. Also by Let X[Y] to be End-completely-regular. To this aim, we need the following result due to Fan ) = End(X)[End(Y)], where End(X)[End(Y)] is the wreath product of the monoids End(X) and End(Y).Let X and Y be two K3-free connected graphs such that girth(X) or girth(Y) is odd. In is End-regular. Here we prove that if X is an End-completely-regular graph and Y is an unretractive graph, then X[Y] is End-completely-regular.Let odd. In , Fan proX and Y be two K3-free connected graphs with girth(X) or girth(Y) being odd, and assume thatX\u2009\u2009is End-completely-regular,Y is unretractive.Then X[Y] is End-completely-regular.Let X and Y be two graphs satisfying the assumptions. To show that X[Y] is End-completely-regular, we prove that for any F \u2208 End(X[Y]), there exists an idempotent endomorphism G \u2208 End(X[Y]) such that \u03c1F = \u03c1G and IF = IG.Let F \u2208 End(X)[End(Y)]. Since End(X[Y]) = End(X)[End(Y)], F = for some s \u2208 End(X) and f \u2208 End(Y)V(X). Thus, for any u \u2208 V(X), there exists fu = f(u) \u2208 End(Y). Let X and Y be K3-free connected graphs with girth(X) or girth(Y) being odd. By u \u2208 V(X), F(Yu)\u2286Yv for some v \u2208 V(X). Note that Y is unretractive. Then F(Yu) = Yv. Since X is End-completely-regular and s \u2208 End(X), by t \u2208 Idpt(X) such that \u03c1t = \u03c1s and It = Is. Clearly, Is is an induced subgraph of X. Hence IF = Is[Y] is an induced subgraph of X[Y].Let X is End-completely-regular, s|Is is an automorphism of Is. Thus for any u \u2208 Is, there exists only one vertex u1 \u2208 Is such that s(u1) = u. Then F(Yu1) = Yu. Now for any \u2208 IF, there exists only one vertex \u2208 IF such that F) = . Define a mapping G from V(X[Y]) to itself in the following way. If \u2208 V(IF), then G) = ; if \u2209 V(IF), then F) = for some \u2208 V(IF). Now let G) = , where is the only vertex in V(IF) such that F) = . Then it is easy to see that G is well-defined. Let \u2208 V(X[Y]). If \u2208 V(IF), then x \u2208 Is. Thus t(x) = x. Hence G) = \u2208 Yt(x). If \u2209 V(IF), then t(x) = u1. Hence G) = \u2208 Yt(x). Therefore, G) \u2208 Yt(x) for any \u2208 V(X[Y]).Since x1, y1), \u2208 V(X[Y]) be such that {, } \u2208 E(X[Y]). If , \u2208 V(IF), then {G), G)} = {, } \u2208 E(X[Y]). If \u2208 V(IF) and \u2209 V(IF), then x1 \u2260 x2 and {x1, x2} \u2208 E(X). Thus G) \u2208 Yt(x1) and G) \u2208 Yt(x2). Since t \u2208 Idpt(X) and {x1, x2} \u2208 E(X), {t(x1), t(x2)} \u2208 E(X). Hence {G), G)} \u2208 E(X[Y]). If \u2209 V(IF) and \u2209 V(IF), there are two cases.Let . Then G) \u2208 Yt(x1) and G) \u2208 Yt(x2). Since t \u2208 Idpt(X) and {x1, x2} \u2208 E(X), {t(x1), t(x2)} \u2208 E(X). Hence we have {G), G)} \u2208 E(X[Y]).Case 2. Assume that x1 = x2 and {y1, y2} \u2208 E(Y). Then we have , \u2208 Yx1 and G), G) \u2208 Yt(x1). Since G|Yx1 is an isomorphism from Yx1 to Yt(x1), {G), G)} \u2208 E(X[Y]). Therefore, G \u2208 End(X[Y]).x, y) \u2208 V(IF), then G2) = G) = . If \u2209 V(IF), then G) \u2208 V(IF). Thus G2) = G)) = G). Hence G \u2208 Idpt(X[Y]). Clearly, IG = IF.If ]\u03c1F = {, ,\u2026, } for some , ,\u2026, \u2208 V(X[Y]). In fact, it is easy to prove that \u03c1F\u2286\u03c1G\u2286\u03c1F. Let \u03c1F. Then, by the definition of G, we have G) = G) = for some \u2208 V(IF) with F) = F = F). So \u03c1G and thus \u03c1F\u2286\u03c1G\u2286\u03c1F. Hence \u03c1F = \u03c1G.Suppose [ such that If = R. Let g \u2208 End(R[Y]). Since R[Y] is not End-completely-regular, there exists g \u2208 End(R[Y]) such that g is not completely regular. By g|Ig is not an automorphism of Ig. Thus there exist x1, x2 \u2208 Ig with x1 \u2260 x2 such that g(x1) = g(x2). Define a mapping F from V(X[Y]) to itself byF \u2208 End(X[Y]) and IF = R[Y]. Now it is easy to see that gF \u2208 End(X[Y]) and IgF = Ig. It follows from (gF)(x1) = (gF)(x2) that (gF)|IgF is not an automorphism of IgF. Hence X[Y] is not End-completely-regular.Let X and Y be two graphs. If at least one of X and Y is not End-completely-regular, then X \u222a Y is not End-completely-regular (where X \u222a Y is the disjoint union of X and Y).Let X is not End-completely-regular. By f \u2208 End(X) such that f|If is not an automorphism of If. Define a mapping F from V(X \u222a Y) to itself byF \u2208 End(X \u222a Y). Now it is easy to see that IF = If \u222a Y and F(x) = f(x) for any x \u2208 V(X). Since f|If is not an automorphism of If, F|IF is not an automorphism of IF. Hence X \u222a Y is not End-completely-regular.Without loss of generality, we may suppose that X and Y be two bipartite graphs. Then X[Y] is End-completely-regular if and only ifX = K1 and Y is End-completely-regular orX is End-completely-regular and Y = K1 or K2.Let Sufficiency. Since K1[Y] = Y and X[K1] = X, we have immediately that K1[Y](X[K1]) is End-completely-regular if and only if Y(X) is End-completely-regular. If X = Y = K2, then X[Y] = K4. Thus End(X[Y]) is a group. Since any group is a completely regular semigroup, X[Y] is End-completely-regular. If X = 2K1 and Y = K2, then X[Y] = 2K2. By X[Y] is End-completely-regular for the following cases (see ases see .Case 1. X = P2 and Y = K2. Let f \u2208 End(X[Y]). If [x1]\u03c1f = {x1}, then [x2]\u03c1f = {x2}. Otherwise, f(x2) = f(z1) or f(x2) = f(z2). Without loss of generality, we can suppose f(x2) = f(z1). Since z1 is adjacent to every vertex of {z2, y1, y2} and {x1, x2} \u2208 E(X[Y]), f(z1) is adjacent to every vertex of {f(z2), f(y1), f(y2), f(x1)}. Note that there is no vertex in X[Y] adjacent to 4 vertices. This is a contradiction. Hence f \u2208 Aut(X[Y]) and so f is completely regular. If [x1]\u03c1f \u2260 {x1}, then f(x1) = f(z1) or f(x1) = f(z2). Without loss of generality, we can suppose f(x1) = f(z1). Then f(x2) = f(z2). Otherwise, a similar argument as above will show that f(x1) is adjacent to every vertex of {f(x2), f(y1), f(y2), f(z2)}, which yield a contradiction. Thus If\u2245K4. Since any endomorphism f maps a clique to a clique of the same size, f(If) = If. By f is completely regular. Hence P2[K2] is End-completely-regular.Case 2. X = K1 \u222a K2 and Y = K2. Let f \u2208 End(X[Y]). If [c1]\u03c1f = {c1}, then [c2]\u03c1f = {c2}. Otherwise, f(c2)\u2208{f(a1), f(a2), f(b1), f(b2)}. Without loss of generality, we can suppose f(c2) = f(a1). Since any endomorphism f maps a clique to a clique of the same size and there is only one clique of size 4 in X[Y], {f(a1), f(a2), f(b1), f(b2)} = {a1, a2, b1, b2}. Note that {c1, c2} \u2208 E(X[Y]). Then {f(c1), f(c2)} = {f(c1), f(a1)}\u2208\u2009E(X[Y]). Thus f(c1)\u2208{f(a1), f(a2), f(b1), f(b2)}, which is a contradiction. Clearly, [x]\u03c1f = {x} for any x \u2208 {a1, a2, b1, b2}. Hence f \u2208 Aut(X[Y]) and so f is completely regular. If [c1]\u03c1f \u2260 {c1}, then f(c1) = f(t) for some t \u2208 {a1, a2, b1, b2}. Without loss of generality, we can suppose f(c1) = f(a1). Then f(c2)\u2208{f(a2), f(b1), f(b2)}. Thus If\u2245K4. Hence f(If) = If and f is completely regular. Consequently, (K1 \u222a K2)[K2] is End-completely-regular.Case 3. X = 2K2 and Y = K2. Let f \u2208 End(X[Y]). If [x]\u03c1f = {x} for any x \u2208 V(X[Y]), then f \u2208 Aut(X[Y]) and so f is completely regular. If f(x) = f(y) for some x, y \u2208 V(X[Y]) with x \u2260 y, without loss of generality, we can suppose f(a1) = f(c1). Since b1, b2, c1, c2 is a clique of size 4 in X[Y], f(b1), f(b2), f(c1), f(c2) is also a clique of size 4 in X[Y]. Note that a2, d1, d2 are adjacent to a1. Then f(a2), f(d1), f(d2) are adjacent to f(a1) = f(c1). Thus f(a2), f(d1), f(d2)\u2208{f(b1), f(b2), f(c2)} and If\u2245K4. Hence f(If) = If and f is completely regular. Consequently, (2K2)[K2] is End-completely-regular. Necessity. We only need to show that X[Y] is not End-completely-regular in the following cases.Case 1\u2009\u2009(X = K2). Then X[Y] = Y + Y. By K2[Y] is not End-completely-regular for the corresponding Y.Case 2\u2009\u2009(X = P2). Then K2 is a retract of X. Since K2[Y] is not End-completely-regular for Y = P2, 2K1, 2K2, K1 \u222a K2, by P2[Y] is not End-completely-regular for the corresponding Y.Case 3\u2009\u2009(X = 2K1). Then X[Y] = 2Y. If Y is bipartite, then X[Y] is also bipartite. By K1)[Y] is not End-completely-regular for the corresponding Y.Case 4\u2009\u2009(X = 2K2). Then X[Y] = 2(Y + Y). Since Y + Y is not End-completely-regular for Y = P2, 2K1, 2K2, K1 \u222a K2, by K2)[Y] is not End-completely-regular for the corresponding Y.Case 5\u2009\u2009(X = K1 \u222a K2). Then X[Y] = Y \u222a (Y + Y). Since Y + Y is not End-completely-regular for Y = P2, 2K1, 2K2, K1 \u222a K2, by K1 \u222a K2)[Y] is not End-completely-regular for the corresponding Y.X and Y under which X[Y] is End-inverse.Next we start to seek the conditions for a lexicographic product of bipartite graphs X and Y be two graphs. If X[Y] is End-inverse, then both X and Y are End-inverse.Let X[Y] is End-inverse, X[Y] is End-regular. By X and Y are End-regular. To show that X is End-inverse, we only need to prove that the idempotents of End(X) commute.Since f1 and f2 be two idempotents in End(X). Define two mappings g1 and g2 from V(X[Y]) to itself byg1 and g2 are two idempotents of End(X[Y]) and so g1g2 = g2g1, since X[Y] is End-inverse. For any \u2208 V(X[Y]), we havef1f2 = f2f1. Hence X is End-inverse.Let f3 and f4 be two idempotents in End(Y). Define two mappings g3 and g4 from V(X[Y]) to itself byg3 and g4 are two idempotents of End(X[Y]) and so g3g4 = g4g3, since X[Y] is End-inverse. For any \u2208 V(X[Y]), we havef3f4 = f4f3. Hence Y is End-inverse, as required.Similarly, let The next theorem characterizes the End-inverse lexicographic products of bipartite graphs.X and Y be two bipartite graphs. Then X[Y] is End-inverse if and only if X[Y] is one of K1[K1], K1[K2], K2[K1], and K2[K2].Let Necessity. This follows directly from Sufficiency. It is easy to see that K1[K1] = K1, K1[K2] = K2[K1] = K2, and K2[K2] = K4 are End-inverse, since they are unretractive."} +{"text": "The aromatic rings attached to the SO 14H13NO5S, was synthesized via a nucleophilic substitution reaction between 3,5-di\u00admethyl\u00adphenol and 2-nitro\u00adbenzene\u00adsulfonyl chloride. The aromatic rings attached to the SO3 group are oriented in a gauche fashion around the ester S\u2014O bond, with a C\u2014S\u2014O\u2014C torsion angle of 84.68\u2005(11)\u00b0. The mol\u00adecules form centrosymmetric dimers via \u03c0\u2013\u03c0 stacking inter\u00adactions between 3,5-di\u00admethyl\u00adphenyl groups (centroid\u2013centroid distance = 3.709\u2005\u00c5). An inter\u00admolecular S=O\u22efN inter\u00adaction between the sulfonyl and nitro groups, with an O\u22efN distance of 2.9840\u2005(18)\u2005\u00c5, organizes the dimers into columns extending along [011]. These columns are further assembled into (111) layers through C\u2014H\u22efO inter\u00adactions.The title compound, C Owing to steric hindrance between the ortho substituents of the benzene ring, the nitro group is twisted relative to the benzene best plane by 39.91\u2005(2)\u00b0, so that the shortest contact of 2.7941\u2005(16)\u2005\u00c5 between the O atoms of these groups is close to the sum of the van der Waals radii.In the title mol\u00adecule Fig.\u00a02, the O1=via inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions between the relatively electron-rich C7\u2013C12 benzene rings layers. The geometry of these inter\u00adactions is given in Table\u00a01o-nitro\u00adaryl\u00adsulfonyl moiety bonded to an aromatic ring through an ester linkage. These are CSD refcodes FEMQUK \u2013103.43\u2005(13)\u00b0. In each of these structures there are either intra- or inter\u00admolecular S=O\u22efN inter\u00adactions between the sulfonate and o-nitro groups.The Cambridge Structural Database portion-wise. The mixture was stirred at 273\u2005K for 30 mins and then at room temperature for 24\u2005h. The product precipitated from the reaction mixture after sitting at 277\u2005K for two weeks. The product was redissolved in di\u00adchloro\u00admethane, and the solvent was allowed to evaporate slowly to give large block-shaped crystals that were suitable for analysis by X-ray diffraction (m.p. 374\u2013378\u2005K).3,5-Di\u00admethyl\u00adphenol was dissolved in chilled di\u00adchloro\u00admethane (25\u2005ml). This was followed by the addition of pyridine . The resulting solution was cooled in an ice bath under an NUiso(H) = 1.2Ueq(C) for CH groups and 1.5Ueq(C) for methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015015078/gk2639sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015015078/gk2639Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015015078/gk2639Isup3.cmlSupporting information file. DOI: 1418463CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of a binuclear mono\u00adcarboxyl\u00adato dirhenium(III) complex with a fulvalene derivative is reported. This compound represents a radical cation salt containing a cluster unit with rhenium\u2013rhenium quadruple bond. 10H8S8)[Re2Br6(CH3COO)]\u00b70.5C2H3Cl3, contains one bis\u00ad(ethyl\u00adenedi\u00adthio)\u00adtetra\u00adthia\u00adfulvalene (ET) radical cation, one \u03bc2-acetato-bis\u00ad[tri\u00adbromido\u00adrhenate(III)] anion and a 1,1,2-tri\u00adchloro\u00adethane mol\u00adecule with half-occupancy disordered about a twofold rotation axis. The tetra\u00adthia\u00adfulvalene fragment adopts an almost planar configuration typical of the ET radical cation. The C atoms of both ethyl\u00adenedi\u00adthio fragments in the cation are disordered over two orientations with occupancy factors 0.65:0.35 and 0.77:0.23. In the anion, six Br atoms and a \u03bc2-acetate ligand form a strongly distorted cubic O2Br6 coordination polyhedron around the Re2 dinuclear centre. In the crystal, centrosymmetrically related ET cations and Re2O2Br6 anions are linked into dimers by \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid-to-centroid distance = 3.826\u2005(8)\u2005\u00c5] and by pairs of additional Re\u22efBr contacts [3.131\u2005(3)\u2005\u00c5], respectively. The dimers are further packed into a three-dimensional network by non-directional inter\u00adionic electrostatic forces and by C\u2014H\u22efBr and C\u2014H\u22efS hydrogen bonds. The disordered 1,1,2-tri\u00adchloro\u00adethane mol\u00adecules occupy solvent-accessible channels along the b axis.The asymmetric unit of the title salt, (C Neither acetic acid nor acetate was used in the synthesis of this radical cation salt. Evidently, the acetate ligand arose by hydrolysis of CH3CN ] anions and 1,1,2-tri\u00adchloro\u00adethane mol\u00adecules in the stoichiometric molar ratio of 1:1:0.5. The solvent mol\u00adecule is disordered over two orientations of equal occupancy about a twofold rotation axis inter\u00adsecting the mid-point of the C\u2014C ethane bond. The tetra\u00adthia\u00adfulvalene fragment adopts an almost planar configuration (r.m.s. deviation = 0.033\u2005\u00c5) that is typical for ET radical cations. The dihedral angle between the five-membered rings is 0.3\u2005(6)\u00b0. The carbon atoms of both ethyl\u00adenedi\u00adthio fragments (C4/C5 and C9/C10) are disordered over two sets of sites with occupancy ratios of 0.65:0.35 and 0.77:0.23, respectively.The title compound Fig.\u00a01 consistsIII atom is coordinated by three Br atoms forming ReBr3 units which are linked by a Re\u2014Re multiple bond [2.2174\u2005(10)\u2005\u00c5] and a bridging \u03bc2-acetate ligand, forming a strongly distorted cubic O2Br6 coordination polyhedron around the Re2 core. The length of the Re\u2014Re bond is very close to the mean value of 2.222\u2005\u00c5 for quadruple bonds \u20132.2731\u2005(9)\u2005\u00c5 for compounds with no bridging ligands and in the range 2.2168\u2005(8)\u20132.2532\u2005(2)\u2005\u00c5 for compounds with O,O-bridging ligands \u2005\u00c5]. Cationic and anionic dimers are packed into a three-dimensional network by non-directional inter\u00admolecular electrostatic forces and by C\u2014H\u22efBr and C\u2014H\u22efS hydrogen bonds -hexa\u00adchlorido\u00addirhenate anion exhibiting the same structure of the title compound anion in a two-electrode U-shaped glass cell with platinum electrodes. The initial current intensity of 0.1\u2005\u00b5A was increased by 0.05\u2005\u00b5A per day to a final value of 0.45\u2005\u00b5A. A mixture of 1,1,2-tri\u00adchloro\u00adethane/aceto\u00adnitrile (12:1 v/v) was used as solvent. [(C4H9)4N]2[Re2Br8] (0.008\u2005mol\u2005l\u22121) was used as electrolyte. After a period of 6\u20137 weeks, black shiny plate-shaped crystals of the title salt suitable for X-ray analysis were formed.The synthesis of the radical cation title salt was performed by galvanostatic anodic oxidation of ET (0.002\u2005mol\u2005lUiso(H) = 1.2Ueq(C) or 1.5Ueq(C) for methyl H atoms. The 1,1,2-tri\u00adchloro\u00adethane mol\u00adecule is disordered over two sets of sites about a twofold rotation axis with equal occupancy. The C4\u2013C5 and C9\u2013C10 groups of the ET cations are disordered over two orientations with occupancy factors of 0.65/0.35 and 0.77/0.23, respectively. These occupancies were initially obtained as free variables by the full-matrix refinement, and were then fixed in the final refinement cycles. The C\u2014C and C\u2014Cl bond lengths in the solvent mol\u00adecule were constrained to be 1.52\u2005(1) and 1.80\u2005(1)\u2005\u00c5, respectively, and the C\u2014Cl bonds of the solvent mol\u00adecule were restrained to have the same lengths to within 0.01\u2005\u00c5. The C\u2014S and C\u2014C bonds of the disordered fragments of the ET cation were also restrained to have the same lengths to within 0.005\u2005\u00c5. The atoms of each disordered fragment, including the solvent mol\u00adecule, were restrained to have approximately the same displacement parameters to within 0.02\u20130.04\u2005\u00c52. DELU restraints to within 0.01\u2005\u00c52 were applied to atoms C4B, C5B, C9B, C10B, C1S and Cl2S. In addition, all non-hydrogen atoms of the solvent mol\u00adecule were restrained to be approximately isotropic to within 0.03\u20130.06\u2005\u00c52. Several outlier reflections (67) that were believed to be affected by the contribution of several unresolved minor twin domains were omitted from the final cycles of refinement, reducing the R factor from 0.061 to 0.052. Attempts to refine the structure using a two-component twin model were unsuccessful. Moreover, the crystals of the title compound are stable but show a strong tendency to splicing. The poor quality of the available crystal may account for the rather low bond precision of the C\u2014C bonds and the presence of several large residual density peaks.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016006058/rz5188sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016006058/rz5188Isup2.hklStructure factors: contains datablock(s) I. DOI: 1473493CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "One O atom from the adjacent hy\u00addroxy\u00adimino-tosyl\u00adhydrazone ligand bridges the CuII cation, forming the centrosymmetric dimeric complex. The cation is in an overall distorted N2O3 square-pyramidal coordination environment. The methyl\u00adbenzene ring is twisted with respect to the hydrazine fragment, with a dihedral angle of 89.54\u2005(9)\u00b0 between the planes. An intra\u00admolecular C\u2014H\u22efO hydrogen bond occurs. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22efS inter\u00adactions. Weak \u03c0\u2013\u03c0 stacking is also observed between parallel benzene rings of adjacent mol\u00adecules, the centroid\u2013centroid distance being 3.9592\u2005(17)\u2005\u00c5.In the title compound, [Cu DOI: 10.1107/S1600536814016651/xu5805Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814016651/xu5805fig1.tifx y z . DOI: x\u00a0+\u00a01,-y\u00a0+\u00a01,-z\u00a0+\u00a01The mol\u00adecular structure of the title compound with labeling and displacement ellipsoids drawn at the 40% probability level showing the dimeric structure. Symmetry code: (i)-Click here for additional data file.10.1107/S1600536814016651/xu5805fig2.tifb . DOI: b-axis showing the column of the aromatic rings with very weak \u03c0\u2013\u03c0 inter\u00adactions.Mol\u00adecules of the title compound arranged along 1014769CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II ions through eight- and 12-membered rings, respectively, forming polymeric chains running along the a-axis direction.The asymmetric unit of the title zinc(II) coordination polymer contains two 4-nitro\u00adbenzoate (NB) anions and two nicotinamide (NA) ligands. Only one of the two NB anions and one of the two NA ligands bridge adjacent Zn 7H4NO4)2(C6H6N2O)2]n, contains two 4-nitro\u00adbenzoate (NB) anions and two nicotinamide (NA) ligands. The ZnII atom has a slightly distorted octa\u00adhedral coordination sphere. In the equatorial plane, it is coordinated by three carboxyl\u00adate O atoms of the NB anions and one O atom of one of the two NA ligands. The axial positions are occupied by the pyridine N atoms of the two NA ligands. In the two NB anions, the carboxyl\u00adate groups are twisted away from the attached benzene rings by 13.8\u2005(2) and 13.4\u2005(2)\u00b0, while the benzene rings are oriented at a dihedral angle of 11.5\u2005(2)\u00b0. The dihedral angle between the NA rings is 10.3\u2005(1)\u00b0. Only one of the two NB anions and one of the two NA ligands bridge adjacent ZnII ions through eight- and twelve-membered rings, respectively, forming polymeric chains running along the a-axis direction. In the crystal, N\u2014H \u22ef O hydrogen bonds link adjacent chains, enclosing R(16), R22(20) and R66(16) ring motifs, forming layers parallel to (01-1). The layers are linked via a number of C\u2014H\u22efO hydrogen bonds, forming a three-dimensional network.The asymmetric unit of the title coordination polymer, [Zn(C N,N-di\u00adethyl\u00adnicotinamide (DENA) is an important respiratory stimulant is one form of niacin. A deficiency of this vitamin leads to loss of copper from the body, known as pellagra disease. Victims of pellagra show unusually high serum and urinary copper levels , 5.855\u2005(4), 4.480\u2005(3)\u2005\u00c5 and 4.67\u2005(6), 3.668\u2005(4), 4.256\u2005(4)\u2005\u00c5, respectively ; see Fig.\u00a03The asymmetric unit of the title polymeric compound contains two 4-nitro\u00adbenzoate (NB) anions and two nicotinamide (NA) ligands; the NB anions act as monodentate ligands Fig.\u00a01. Only onon Fig.\u00a02. In the II cation form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two pyridine N atoms (N3 and N5) of the two NA ligands in the axial positions of the three NB anions and one O atom (O10) of one of the two NA ligands in the equatorial plane around the Zns Table\u00a01.carboxyl\u00adate and Zn\u2014N distances are 2.147\u2005(2)\u2005\u00c5 and 2.285\u2005(3)\u2005\u00c5, respectively, while Zn1\u2014O10 distance is 2.280\u2005(2)\u2005\u00c5. The Zn1 atom lies 1.4330\u2005(4)\u2005\u00c5 and 0.1897\u2005(4)\u2005\u00c5 above the planar (O1/O2/C1) and (O5/O6/C8) carboxyl\u00adate groups, respectively. The average O\u2014Zn\u2014O and O\u2014Zn\u2014N bond angles are 89.93\u2005(10) and 89.99\u2005(10)\u00b0, respectively.The near equality of the C1\u2014O1 [1.247\u2005(4)\u2005\u00c5], C1\u2014O2 [1.261\u2005(4)\u2005\u00c5], C8\u2014O5 [1.248\u2005(4)\u2005\u00c5] and C8\u2014O6 [1.255\u2005(4)\u2005\u00c5] bonds in the carboxyl\u00adate groups indicate delocalized bonding arrangements, rather than localized single and double bonds. The average Zn\u2014OA (C2\u2014C7) and B (C9\u2014C14)] are 13.8\u2005(2) and 13.4\u2005(2)\u00b0, respectively, while the benzene rings are oriented at a dihedral angle of 11.5\u2005(2)\u00b0. The dihedral angle between the nicotin\u00adamide rings [C (N3/C15\u2014C19) and D (N5/C21\u2014C25)] is 10.3\u2005(1)\u00b0, and they are oriented with respect to benzene rings A and B at dihedral angles of A/C = 17.3\u2005(1), A/D = 7.7\u2005(1), B/C = 28.8\u2005(1) and B/D = 18.9\u2005(1)\u00b0.The dihedral angles between the planar carboxyl\u00adate groups [(O1/O2/C1) and (O5/O6/C8)] and the adjacent benzene rings [c (c = carboxylate) and N\u2014H\u22efOn (n = nicotinamide) hydrogen bonds (Table\u00a02R(16), et al., 1995n\u22efOc and inter\u00admol\u00adec\u00adular C\u2014Hn\u22efOnb (nb = nitro\u00adbenzoate) and C\u2014Hn\u22efOn hydrogen bonds Fig.\u00a04. Weak ins Table\u00a01 link the4\u00b7H2O in H2O (25\u2005ml) and nicotinamide in H2O (25\u2005ml) with sodium 4-nitro\u00adbenzoate in H2O (150\u2005ml). The mixture was filtered and set aside to crystallize at ambient temperature for one week, giving yellow block-like crystals.The title compound was prepared by the reaction of ZnSOUiso(H) = 1.2Ueq. The highest residual electron density and the deepest hole were found 0.29\u2005\u00c5 and 0.48\u2005\u00c5 from atoms N6 and Zn1, respectively.The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0310.1107/S2056989015006490/su5099sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015006490/su5099Isup2.hklStructure factors: contains datablock(s) I. DOI: 1057121CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H,3H,5H)-trione, which crystallized with two mol\u00adecules in its asymmetric unit along with two solvent water mol\u00adecules. A hydrogen-bonded sheet is formed by a combination of N\u2014H\u22efO and Owater\u2014H\u22efO hydrogen bonds, which are further inter\u00adconnected by C\u2014H\u22ef\u03c0ar\u00adyl inter\u00adactions, leading to a three-dimensional supra\u00admolecular architecture.Reaction of 4-methyl benzyl chloride with barbituric acid gave 5,5-bis\u00ad(4-methyl\u00adbenz\u00adyl)pyrimidine-2,4,6, with similar conformations and two independent water mol\u00adecules. In the crystal, N\u2014H\u22efO and Owater\u2014H\u22efO hydrogen bonds link all moieties into two crystallographically independent kinds of sheets parallel to the ac plane. These independent sheets, each containing either A or B mol\u00adecules, are further alternately stacked along the b axis and inter\u00adconnected via C\u2014H\u22ef\u03c0ar\u00adyl inter\u00adactions.The asymmetric unit of the title compound, C In B, the corresponding angles are 50.44\u2005(18), 69.90\u2005(19) and 59.8\u2005(2)\u00b0, respectively. In the related compound 5,5-di\u00adbenzyl\u00adbarbituric acid monohydrate (II) via strong N2\u2014HN2\u22efO4 (A) and N3\u2014HN3\u22efO7 (B) hydrogen bonds C(6). Six chains pass through the unit cell. These chains are linked via water mol\u00adecules through N1\u2014H1\u22efO1 and O1\u2014H1B\u22efO5 (for A) and N4\u2014HN4\u22efO2 and O2\u2014H2A\u22efO8 (for B) hydrogen bonds (Table\u00a01D(2). In addition, the symmetry-dependent parallel chains are inter\u00adconnected via bridging water mol\u00adecules through O\u2014H\u22efO3 and O1\u2014HIB\u22efO5 (for A) and O2\u2014H2B\u22efO6 and O2\u2014H2A\u22efO8 (for B) hydrogen bonds crystallizes in the monoclinic space group P21/n. Secondly, (I)There are several inter\u00adesting differences between the two chemically closely related structures (I)To an ice-cooled stirring solution of acetonitrile (5\u2005ml), 4-methyl benzyl chloride , 1,8-diazabicycloundec-7-ene (DBU) and barbituric acid were added. The reaction mixture was stirred to the room temperature and then refluxed for 8\u2005h. Thin-layer chromatography showed the absence of any starting material. The reaction mixture was cooled and poured into ice-cold water. The solid obtained was extracted with ethyl acetate and the organic layer was washed with saturated ammonium chloride solution and dried over anhydrous sodium sulphate. The solvent was removed under reduced pressure to give the title compound as a white solid .Colourless prisms of the title compound suitable for diffraction studies were grown from an ethyl acetate\u2013petroleum ether solvent system in the ratio 2.5:7.5, by the solvent evaporation technique.Ueq(C) for methyl H atoms and = 1.2Ueq for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901402619X/cv5478sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901402619X/cv5478Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901402619X/cv5478Isup3.cmlSupporting information file. DOI: 1036677CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecular components, enclosing In the title salt, [Zn(C 8H4NO2)(C6H6N2O)(H2O)3](C8H4NO2), contains one complex cation and one 4-cyano\u00adbenzoate (CNB) counter-anion. The ZnII atom in the cation is coordinated by one 4-cyano\u00adbenzoate ligand, one nicotinamide (NA) ligand and three water mol\u00adecules, the CNB anion thereby coordinating in a bidentate O,O\u2032-mode through the carboxyl\u00adate group. The latter, together with one water O atom and the N atom of the NA ligand, form a distorted square-planar arrangement, while the considerably distorted octa\u00adhedral coordination sphere of the ZnII atom is completed by the two O atoms of additional water mol\u00adecules in the axial positions. The dihedral angles between the planar carboxyl\u00adate groups and the adjacent benzene rings in the two anions are 10.25\u2005(10) and 5.89\u2005(14)\u00b0. Inter\u00admolecular O\u2014H\u22efO hydrogen bonds link two of the coordinating water mol\u00adecules to two free CNB anions. In the crystal, further hydrogen-bonding inter\u00adactions are present, namely N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds that link the mol\u00adecular components, enclosing R22(12), R33(8) and R33(9) ring motifs and forming layers parallel to (001). \u03c0\u2013\u03c0 contacts between benzene rings [centroid-to-centroid distances = 3.791\u2005(1) and 3.882\u2005(1)\u2005\u00c5] may further stabilize the crystal structure.The asymmetric unit of the title salt, [Zn(C N,N-di\u00adethyl\u00adnicotinamide (DENA), an important respiratory stimulant , one form of niacin (C6H6ON2)(H2O)3](C8H4O2N), is composed of one complex cation and one 4-cyano\u00adbenzoate (CNB) counter-anion. The ZnII atom is coordinated by one 4-cyano\u00adbenzoate (CNB) anion, one nicotinamide (NA) ligand and three water mol\u00adecules, the CNB anion and NA ligand coordinating in bidentate and monodentate modes, respectively in the axial positions around the Zn1 atom show a distorted square-planar arrangement, while the considerably distorted octa\u00adhedral coordination environment of Zns Table\u00a01.9H10NO2)(C6H6N2O)\u00b72H2O] (C6H6N2O)]\u00b7H2O 2(C6H6N2O)] \u2005\u00c5], C1\u2014O2 [1.2591\u2005(19)\u2005\u00c5] and C15\u2014O7 [1.266\u2005(2)\u2005\u00c5], C15\u2014O8 [1.237\u2005(2)\u2005\u00c5] bonds in the carboxyl\u00adate groups indicate delocalized bonding arrangements, rather than localized single and double bonds. The average Zn\u2014O bond lengths are 2.19\u2005(11)\u2005\u00c5 for benzoate oxygen atoms and 2.10\u2005(9)\u2005\u00c5 for water oxygen atoms; the Zn\u2014N bond length is 2.0545\u2005(12)\u2005\u00c5, close to the values in related structures. The Zn1 atom lies 0.0093\u2005(2)\u2005\u00c5 above the planar (O1/O2/C1) carboxyl\u00adate group, with a bite angle of 59.48\u2005(4)\u00b0. Corresponding O\u2014Zn\u2014O angles are 60.03\u2005(6)\u00b0 in [Zn(CA (C2\u2013C7) and C (C16\u2013C21)] are 10.25\u2005(10) and 5.89\u2005(14)\u00b0, respectively, while the benzene rings and benzene and pyridine [B (N2/C9\u201413)] rings are oriented at dihedral angles of A/C = 77.84\u2005(6), A/B = 8.97\u2005(5) and B/C = 71.43\u2005(5)\u00b0.The dihedral angles between the planar carboxyl\u00adate groups [(O1/O2/C1) and (O7/O8/C15)] and the adjacent benzene rings , may further stabilize the structure, with centroid-to-centroid distances of 3.791\u2005(1)\u2005\u00c5 and 3.882\u2005(1)\u2005\u00c5, respectively.In the crystal, N\u2014H\u22efOs Table\u00a02 link the4\u00b77H2O in H2O (30\u2005ml) and nicotinamide in H2O (50\u2005ml) with sodium 4-cyano\u00adbenzoate in H2O (100\u2005ml). The mixture was filtered and set aside to crystallize at ambient temperature for several days, giving colourless single crystals.The title compound was prepared by the reaction of ZnSO2 group) and H41, H42, H51, H52, H61 and H62 (as part of the water mol\u00adecules) were located in a difference Fourier map and were refined freely. The aromatic C-bound H atoms were positioned geometrically with C\u2014H = 0.93\u2005\u00c5, and constrained to ride on their parent atoms, with Uiso(H) = 1.2Ueq(C).The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0310.1107/S2056989015009743/wm5158sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015009743/wm5158Isup2.hklStructure factors: contains datablock(s) I. DOI: 1401948CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports6: Article number: 2019910.1038/srep20199; published online: 02012016; updated: 04202016.This Article contains typographical errors in in the Methods section.K\u03b1 radiation was used with the X-ray source that was run at 14.5\u2009kV. The energy analyzer operated at a constant pass energy of 50\u2009eV. Since the XPS detectable depth for Fe 2p and Pt 4f electrons are only 5.19\u2009nm and 4.17\u2009nm43, the NiTi(Nb)-SMA/L10-FePt(3\u2009nm)/Ta(5\u2009nm) samples with different elastic strain treatments were designed for the XPS tests.\u201d\u201cMg should read:K\u03b1 radiation was used with the X-ray source that was run at 14.5\u2009kV. The energy analyzer operated at a constant pass energy of 50\u2009eV. Since the XPS detectable depth for Fe 2p and Pt 4f electrons are only 3.9\u2009nm and 3.2\u2009nm43, the NiTi(Nb)-SMA/L10-FePt(3\u2009nm)/Ta(5\u2009nm) samples with different elastic strain treatments were designed for the XPS tests.\u201d\u201cAl"} +{"text": "The supra\u00admolecular inter\u00adactions between cations and anions lead to a two-dimensional network structure parallel to (001).The complex cation of the title salt, (C9H6NO3S)2. In its crystal structure, the complex cation [Cd(TEA)2]2+ and two independent NBTA\u2212 units with essentially similar geometries and conformations are present. In the complex cation, each TEA mol\u00adecule behaves as an N,O,O\u2032,O\u2032\u2032-tetra\u00addentate ligand, giving rise to an eight-coordinate CdII ion with a bicapped trigonal\u2013prismatic configuration. All ethanol groups of each TEA mol\u00adecule form three five-membered chelate rings around the CdII ion. The Cd\u2014O and Cd\u2014N distances are in the ranges 2.392\u2005(2)\u20132.478\u2005(2) and 2.465\u2005(2)\u20132.475\u2005(3)\u2005\u00c5, respectively. O\u2014H\u22efO hydrogen bonds between the TEA hy\u00addroxy groups and carboxyl\u00adate O atoms connect cationic and anionic moieties into chains parallel to [110]. Each NBTA\u2212 anion is additionally linked to a symmetry-related anion through \u03c0\u2013\u03c0 stacking inter\u00adactions between the benzene and thia\u00adzoline rings [minimum centroid-to-centroid separation = 3.604\u2005(2)\u2005\u00c5]. Together with additional C\u2014H\u22efO inter\u00adactions, these establish a double-layer polymeric network parallel to (001).The reaction of 2-acetic acid (NBTA) and tri\u00adethano\u00adlamine (TEA) with Cd(CH The N\u2014Cd\u2014O bond angles range from 68.58\u2005(8) to 122.59\u2005(10)\u00b0 and the O\u2014Cd\u2014O angles are in an inter\u00adval of 72.54\u2005(9) to 162.13\u2005(11)\u00b0. Both thia\u00adzoline rings (C1/C6/N1/C7/S1 and C1A/C6A/N1A/C7A/S1A) and bicyclic benzo\u00adthia\u00adzole units (N1/S1/C1\u2013C7 and (N1A/S1A/C1A\u2013C7A) are close to planar, the largest deviations from the least-squares planes being 0.002\u2005(2), 0.004\u2005(2) and 0.008\u2005(3), 0.005\u2005(3)\u2005\u00c5, respectively. The dihedral angles between the plane of the carboxyl\u00adate group and the attached benzo\u00adthia\u00adzole ring system are 77.895\u2005(3) and 71.408\u2005(3)\u00b0 in the two anions.The structure of the mol\u00adecular entities of (I)\u2212 anions into a chain structure extending parallel to [110], whereby each cation is surrounded by four NBTA\u2212 anions. The H atoms of all hydroxyl groups of the TEA ligands form a hydrogen bond with a carboxyl\u00adate O atom of the NBTA\u2212 ions. In addition, there is weak hydrogen bond between one \u2013CH2 group and the O1 atom of the NBTA anion, with a C\u22efO distance of 3.282\u2005(6)\u00c5 and thia\u00adzoline rings (centroid Cg2) of adjacent inversion-related mol\u00adecules [Cg1\u22efCg2 = 3.604\u2005(2)\u2005\u00c5] \u00c5 Table\u00a01. The abofs Fig.\u00a02. The NBT\u00c5] Fig.\u00a03. Togethes-, d-, p-, and f-block elements have been reported. Structures containing the [Cd(TEA)2]2+ cation are deposited in the CSD with reference codes EYIPAD, MEVQIN and YOVBIU.A survey of the Cambridge Structural Database 2 was slowly added an ethano\u00adlic solution (5\u2005ml) containing TEA (132\u2005\u00b5l) and NBT under constant stirring. A bright-yellow crystalline product was obtained at room temperature by solvent evaporation after four weeks. Yield: 75%; calculated for C30H42CdN4O12S2: C, 43.56; H, 5.12; N, 6.77, found: C, 43.61; H, 5.15; N, 6.69To an aqueous solution (2.5\u2005ml) of Cd(CHUiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016004515/wm5280sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016004515/wm5280Isup2.hklStructure factors: contains datablock(s) I. DOI: 1468939CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The corresponding coordination polyhedron is a distorted trigonal bipyramid, with Cd\u2014Cl distances in the range 2.4829\u2005(4)\u20132.6402\u2005(4)\u2005\u00c5. The bipyramids are condensed into a polyanionic zigzag chain extending parallel to [101]. The tetra\u00admethyl\u00adguanidinium cations are situated between the polyanionic chains and are linked to them through N\u2014H\u22efCl hydrogen bonds, forming a layered network parallel to (010).In the structure of the title salt, {(C Several cationic complexes of Pd, Ga and Pt have been reported with tetra\u00admethyl\u00adguanidine acting as a ligand x, y, 1\u00a0\u2212\u00a0z] are in axial positions with a Cl3\u2014Cd1\u2014Cl4i angle of 166.347\u2005(10)\u00b0. The equatorial Cd\u2014Cl bond lengths range from 2.4829\u2005(4)\u2005\u00c5 to 2.5829\u2005(4)\u2005\u00c5 while the axial bond lengths Cd1\u2014Cl3 and Cd1\u2014Cl4i are 2.5854\u2005(4)\u2005\u00c5 and 2.6403\u2005(4)\u2005\u00c5, respectively. The CdCl4 moieties of the asymmetric unit are related by an inversion center, generating an extended zigzag chain of edge-sharing trigonal bipyramids running parallel to [101]. These 1\u221e[CdCl4/2Cl1/1]\u2212 chains are formed by the bridging atoms Cl2, Cl3, Cl4 and Cl4i with a Cd\u2014Cd\u2014Cd angle of 137.893\u2005(6)\u00b0. The corrugation of the chains results in rather short Cd\u22efCd distances of 3.8720\u2005(3) and 3.8026\u2005(3)\u2005\u00c5. The same kind of zigzag chain is found, for example, in the [CdCl3]\u2212 salt obtained with benzyl\u00adtri\u00adethyl\u00adammonium as counter-cation \u00b0 in the structure of the benzyl\u00adtri\u00adethyl\u00adammonium compound compared with 129.859\u2005(2)\u00b0 for the present structure. The tetra\u00admethyl\u00adguanidinium cation has the central atom C1 in an almost trigonal\u2013planar configuration. The three N\u2014C\u2014N angles range from 119.26\u2005(14) to 121.14\u2005(14)\u00b0 and the r.m.s deviation from the least-squares plane calculated with atoms C1, N1, N2 and N3 is only 0.005\u2005\u00c5. The corresponding C\u2014N bond lengths of 1.330\u2005(2), 1.3360\u2005(19), and 1.3441\u2005(19)\u2005\u00c5 indicate a partial double-bond character. Hence the positive charge may be considered as delocalized in the CN3 plane 1\u221e[CdCl4/2Cl1/1]\u2212 chains are inter\u00adconnected through N\u2014H\u22ef\u00b7Cl hydrogen bonds by pairs of tetra\u00admethyl\u00adguanidinium cations linked to symmetry-related Cl1 atoms = 1.5Ueq(C). H atoms bonded to the N atoms were located from a Fourier difference map and were refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015020836/wm5207sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015020836/wm5207Isup2.hklStructure factors: contains datablock(s) I. DOI: 1434977CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked via pairs of O\u2014H\u22efO hydrogen bonds, involving the boronic acid OH groups, forming inversion dimers. The dimers are linked via O\u2014H\u22efO hydrogen bonds, involving a boronic acid OH group and the carbonyl O atom, forming undulating sheets parallel to (10-2). Within the sheets there are also C\u2014H\u22efO hydrogen bonds present, also involving the carbonyl O atom. The sheets are linked via C\u2014H\u22ef\u03c0 and offset face-to-face \u03c0-inter\u00adactions between inversion-related mol\u00adecules , forming a three-dimensional structure.In the title compound, C The unit cell consists of four mol\u00adecular units which form \u03c0-aggregated pairs in a head-to-tail fashion and are stabilized through offset face-to-face \u03c0-inter\u00adactions ; see Fig.\u00a04A of the boronic acid is in an inversion-related hydrogen-bonded network with the oxygen O2 of the boronic acid at a distance of 2.762\u2005(1)\u2005\u00c5, forming a head-to-head hydrogen-bonded network with adjacent mol\u00adecules (2-methyl\u00adimidazol\u00adyl)boronate monohydrate, tris\u00ad(4-(di\u00admethyl\u00adamino)\u00adpyridinium) bis\u00ad(4-(di\u00admethyl\u00adamino)\u00adpyridine) tris\u00ad(4-carb\u00adoxy\u00adbenzene\u00adboronate) trihydrate and 4-acetyl\u00adpyridine oxime 4-carb\u00adoxy\u00adbenzene\u00adboronate dihydrate which presented out-of-plane tilt angles of ca 10.45\u201327.74\u00b0 and O\u2014B\u2014O bond angles of ca 114.23\u2013124.94\u00b0 bis\u00ad(4-carboxyl\u00adato\u00adphenyl\u00adboronic acid) tetra\u00adhydrate, where M is nickel, manganese or cobalt. These structures where similar to the title compound and exhibited similar characteristics for the O\u2014B\u2014O bond angle and the out-of-plane tilt of the carboxyl acid group compared to the title compound. The carb\u00adoxy\u00adlic group deviated from the plane with an angle of ca 3.40\u20134.53\u00b0 and the O\u2014B\u2014O bond angles were in the range of ca 121.43\u2013122.18\u00b0 4-(di\u00adhydroxy\u00adbor\u00adyl)benzoic acid monohydrate exhibited a tilt angle of ca 2.14\u00b0 for the carb\u00adoxy\u00adlic group and an O\u2014B\u2014O bond angle of ca 126.53\u00b0 . The compound was then crystallized from a solution of 1% MeOH in CH2Cl2 layered with hexane to give a single crystal suitable for X-ray diffraction.The compound was purchased from Alfa Aesar and was purified with silica gel column chromatography using CHUiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Uiso(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015015923/su5188sup1.cifCrystal structure: contains datablock(s) I, publication_text. DOI: 10.1107/S2056989015015923/su5188Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015015923/su5188Isup3.cmlSupporting information file. DOI: 1416885CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The grid layers are held together by hydrogen bonds between the organic guest mol\u00adecules and the host framework and gives rise to a three-dimensional supra\u00admolecular architecture.{[Fe(H 2Fe(CN)4(H2O)2]\u00b72C12H10N2}n, the asymmetric unit contains one FeII cation, two water mol\u00adecules, two di\u00adcyanido\u00adargentate(I) anions and two uncoordinating 1,2-bis\u00ad(pyridin-2-yl)ethyl\u00adene mol\u00adecules. Each FeII atom is six-coordinated in a nearly regular octa\u00adhedral geometry by four N atoms from di\u00adcyanido\u00adargentate(I) bridges and two coordinating water mol\u00adecules. The FeII atoms are bridged by di\u00adcyanido\u00adargentate(I) units to give a two-dimensional layer with square-grid spaces. The inter\u00adgrid spaces with inter\u00adlayer distance of 6.550\u2005(2)\u2005\u00c5 are occupied by 2,2\u2032-bpe guest mol\u00adecules which form O\u2014H\u22efN hydrogen bonds to the host layers. This leads to an extended three-dimensional supra\u00admolecular architecture. The structure of the title compound is compared with some related compounds containing di\u00adcyanido\u00adargentate(I) ligands and N-donor organic co-ligands.In the title compound, {[Ag These structural properties lead to potential inter\u00adesting applications in the filed of magnetism, sensing, porous mater\u00adials and catalysis 4(H2O)2]\u00b72C12H10N2}n, (I)One-, two- and three-dimensional frameworks containing di\u00adcyanido\u00adargentate(I) and N-donor linkers such as pyrazine, 4,4\u2032-bpy and 4,4\u2032-bpe ligands have been studied ligands, two water mol\u00adecules and two uncoord\u00adinating 2,2\u2032-bpe mol\u00adecules ligands and two water mol\u00adecules.The asymmetric unit consists of one Fees Fig.\u00a01. Ag1 and\u00c52 Fig.\u00a02. The FeIW\u22efN5 = 2.07\u2005\u00c5 and O2\u2013H4W\u22efN6 = 2.09\u2005\u00c5) , while the other two arrange themselves across the host layer to form also hydrogen bonds (O1\u2014H1W\u22efN7 = 2.14\u2005\u00c5 and O2\u2014H3W\u22efN8 = 2.15\u2005\u00c5) to the host layers. These hydrogen bonds generate an extended three-dimensional supra\u00admolecular framework.Four independent 2,2\u2032-bpe mol\u00adecules are located between adjacent grid layers of which two are parallel (blue) to the grid layers and two non-parallel (red) Fig.\u00a03. The int\u00c5) Fig.\u00a04a, while\u00c5) Fig.\u00a04b to the4[Ag(CN)2]2]n (imH = imidazole), a one-dimensional chain via bridging di\u00adcyanido\u00adargentate(I) is found, while all imidazole mol\u00adecules act as a terminal ligand 2]2]n (3-Fpy = 3-fluoro\u00adpyridine) consists of four cyanide moieties occupying the equatorial positions generating a square grid-type structure similar to that of the title compound, while the axial positions are occupied by two terminal 3-Fpy ligands instead of two water mol\u00adecules in (I)et al., 20072]2].pz}n (pz = pyrazine), [Mn2[Ag(CN)2]2]n, [Fe2[Ag(CN)2]2]n and [Fe(bpe)2[Ag(CN)2]2]n 2] was added dropwise to an MeOH\u2013H2O mixed solution of (NH4)2[Fe(SO4)2]\u00b76H2O and 2,2\u2032-bpe at room temperature. After filtration and slow evaporation for 1\u2005d, yellow crystals were obtained.An aqueous solution (5\u2005ml) of K[Ag(CN)isoU(H) = 1.2eqU(C). Water H atoms were located in difference Fourier maps and refined isotropically.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S1600536814016250/vn2085sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814016250/vn2085Isup2.hklStructure factors: contains datablock(s) I. DOI: 1013775CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In this manuscript the mol\u00adecular and supra\u00admolecular structure of 2\u2032,3,6\u2032-tri\u00adchloro\u00adbiphenyl-2-ylboronic acid tetra\u00adhydro\u00adfuran monosolvate is presented 12H8BCl3O2\u00b7C4H8O, crystallizes as a tetra\u00adhydro\u00adfuran monosolvate. The boronic acid group adopts a syn\u2013anti conformation and is significantly twisted along the carbon\u2013boron bond by 69.2\u2005(1)\u00b0, due to considerable steric hindrance from the 2\u2032,6\u2032-di\u00adchloro\u00adphenyl group that is located ortho to the boronic acid substituent. The phenyl rings of the biphenyl are almost perpendicular to one another, with a dihedral angle of 87.9\u2005(1)\u00b0 between them. In the crystal, adjacent mol\u00adecules are linked via O\u2014H\u22efO inter\u00adactions to form centrosymmetric dimers with R22(8) motifs, which have recently been shown to be energetically very favourable. The hy\u00addroxy groups are in an anti conformation and are also engaged in hydrogen-bonding inter\u00adactions with the O atom of the tetra\u00adhydro\u00adfuran solvent mol\u00adecule. Cl\u22efCl halogen-bonding inter\u00adactions [Cl\u22efCl = 3.464\u2005(1)\u2005\u00c5] link neigbouring dimers into chains running along [010]. Further aggregation occurs due to an additional Cl\u22efCl halogen bond [Cl\u22efCl = 3.387\u2005(1)\u2005\u00c5].The title compound, C In the structure of the related (2-bi\u00adphenyl\u00adyl)boronic acid \u00b0], whereas in (2-bi\u00adphenyl\u00adyl)boronic acid they are rotated by only 48.4 or 45.4\u00b0 for the two unique mol\u00adecules in the asymmetric unit.The B\u2014C [1.5907\u2005(16)\u2005\u00c5] and B\u2014O bonds in the title compound (I)on Fig.\u00a01. The boranti-oriented OH group is engaged in an inter\u00admolecular O\u2014H\u22efO hydrogen bond \u2005\u00c5; the sum of the van der Waals radii for Cl is 3.50\u2005\u00c5]. In terms of geometry of this contact, it can be classified as a type I halogen bond , \u2005\u00c5] .In the crystal, centrosymmetric O\u2014H\u22efO hydrogen-bonded dimers are formed. The d Table\u00a01 with thend Fig.\u00a02a, A solution of 2-iodo-2\u2032,3,6\u2032-tri\u00adchloro\u00adbiphenyl in THF (50\u2005mL) was added to a stirred solution of n-BuLi (10\u2005mmol) in THF (30\u2005mL) at 195\u2005K. The resulting colorless solution was stirred for 1\u2005h to give a colorless precipitate. The electrophile, B(OMe)3 was then added to the stirred mixture to give a colorless solution which was stirred for 1\u2005h and then hydrolyzed with H2O (100\u2005mL). Dilute aq. H2SO4 was added until the pH was slightly acidic. Et2O (50\u2005mL) was next added and the mixture stirred for 10\u2005min. The organic phase was separ\u00adated and the aqueous phase was extracted with Et2O (20\u2005mL). The combined organic solutions were dried over MgSO4 and evaporated to give a colorless precipitate, yield 2.0\u2005g (66%). 1H NMR : \u03b4 = 8.00 , 7.51 , 7.39 , 7.05 ;13C{1H} NMR : \u03b4 = 141.49, 139.62, 139 (br), 136.17, 134.84, 130.49, 129.99, 128.24, 128.14, 127.53.Crystals suitable for X-ray diffraction analysis were grown by slow evaporation of a THF solution.Uiso(phenyl H) = 1.2Ueq(C). The positions of the OH hydrogen atoms were first found in a difference map. Then their bond lengths were restrained in the last least-squares cycles, with an O\u2014H distance of 0.85\u2005\u00c5 and their coordinates refined with Uiso(hydroxyl H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901502054X/sj5482sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901502054X/sj5482Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901502054X/sj5482Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S205698901502054X/sj5482Isup4.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S205698901502054X/sj5482Isup5.cmlSupporting information file. DOI: 1434205CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Eight O\u2014H\u22efO hydrogen bonds [d(O\u22efH) < 2.00\u2005\u00c5], form a two-dimensional network parallel to the ab plane.In the crystal structure of the title compound, [Fe(dmbpy)(H 12H12N2)(H2O)4]SO4, the central FeII ion is coordinated by two N atoms from the 5,5\u2032-dimethyl-2,2\u2032-bi\u00adpyridine ligand and four water O atoms in a distorted octa\u00adhedral geometry. The Fe\u2014O coordination bond lengths vary from 2.080\u2005(3) to 2.110\u2005(3)\u2005\u00c5, while the two Fe\u2014N coordination bonds have practically identical lengths [2.175\u2005(3) and 2.177\u2005(3)\u2005\u00c5]. The chelating N\u2014Fe\u2014N angle of 75.6\u2005(1)\u00b0 shows the largest deviation from an ideal octa\u00adhedral geometry; the other coordination angles deviate from ideal values by 0.1\u2005(1) to 9.1\u2005(1)\u00b0. O\u2014H\u22efO hydrogen bonding between the four aqua ligands of the cationic complex and four O-atom acceptors of the anion leads to the formation of layers parallel to the ab plane. Neighbouring layers further inter\u00adact by means of C\u2014H\u22efO and \u03c0\u2013\u03c0 inter\u00adactions involving the laterally positioned bi\u00adpyridine rings. The perpen\u00addicular distance between \u03c0\u2013\u03c0 inter\u00adacting rings is 3.365\u2005(2)\u2005\u00c5, with a centroid\u2013centroid distance of 3.702\u2005(3)\u2005\u00c5.In the title compound, [Fe(C The FeII atom is in a distorted octa\u00adhedral coordination environment and the equatorial plane of the octa\u00adhedron is formed by a pair of nitro\u00adgen donors from the 5,5\u2032-dimethyl-2,2\u2032-bipyridyl ligand and two mol\u00adecules of water, while the axial sites are occupied by two other water mol\u00adecules. The equatorial donor atoms are nearly coplanar (r.m.s. deviation = 0.0062\u2005\u00c5), while the deviation of the Fe atom from the least-squares plane is somewhat larger [0.021\u2005(2)\u2005\u00c5]. The bi\u00adpyridine chelating angle N1\u2014Fe\u2014N2 of 75.6\u2005(1)\u00b0 shows the most significant deviation from an ideal octa\u00adhedral geometry. The other angular distortions from an ideal octa\u00adhedral geometry are in the range 0.1\u2005(1) to 9.1\u2005(1)\u00b0. The S\u2014O bond lengths [1.466\u2005(3)\u20131.480\u2005(3)\u2005\u00c5] and O\u2014S\u2014O angles [108.8\u2005(2)\u2013109.9\u2005(2)\u00b0] indicate a nearly ideal tetra\u00adhedral geometry for the anion.A mol\u00adecular view of complex (I)2O)4]2+ cations engages all four coordinating water mol\u00adecules in hydrogen bonding to four [SO4]2\u2212 anions . The anions surrounding the cationic unit are positioned at similar Fe\u22efS distance of 4]2\u2212 anions appears surrounded with four cationic units, where its four O atoms engage as acceptors in bifurcated O\u2014H\u22efO hydrogen bonds towards neighbouring cations . Such a mutual arrangement leads to the formation of a two-dimensional hydrogen-bonded network parallel to the ab plane . Laterally arranged aromatic rings of the 5,5\u2032-dimethyl-2,2\u2032-bi\u00adpyridine ligand in neighbouring layers inter\u00adact by means of weak C\u2014H\u22efO and \u03c0\u2013\u03c0 inter\u00adactions, forming the three-dimensional crystal packing in water\u2013ethanol . The mixture was transferred to a Teflon-lined autoclave and heated at 410\u2005K for 3\u2005d. The autoclave was then allowed to cool to ambient temperature. Red crystals of (I)The title compound, (I)Uiso(H) values were equal to 1.2 and 1.5 times Ueq of the corresponding C(sp2) and C(sp3) atoms. The H atoms of the four water mol\u00adecules were initially located in a difference Fourier map. During the refinement, these H atoms were allowed to ride on their parent O atoms and also to rotate about the corresponding Fe\u2014O bonds. The Uiso(H) values were set equal to 1.2 times Ueq of the parent O atom. The reflections (100) and (002) were excluded from the refinement because they were nearly completely obscured by the beamstop.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814024982/vn2087sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S1600536814024982/vn2087Isup2.hklStructure factors: contains datablock(s) I. DOI: 1034106CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The core is decorated by four tri\u00adphenyl\u00adphosphine ligands bonded to the AgI cations. Ag\u22efAg distances as short as 3.133\u2005(9)\u2005\u00c5 suggest the presence of argentophilic (d10\u22efd10) inter\u00adactions.In the title compound a distorted Ag 6H5)3P]Ag}4{NCO}4], a distorted Ag4N4-heterocubane core is set up by four AgI ions being coordinated by the N atoms of the cyanato anions in a \u03bc3-bridging mode. In addition, a tri\u00adphenyl\u00adphosphine ligand is datively bonded to each of the AgI ions. Intra\u00admolecular Ag\u22efAg distances as short as 3.133\u2005(9)\u2005\u00c5 suggest the presence of argentophilic (d10\u22efd10) inter\u00adactions. Five moderate-to-weak C\u2014H\u22efO hydrogen-bonding inter\u00adactions are observed in the crystal structure, spanning a three-dimensional network. A region of electron density was treated with the SQUEEZE procedure in PLATON following unsuccessful attempts to model it as being part of disordered tetra\u00adhydro\u00adfuran solvent mol\u00adecules. The given chemical formula and other crystal data do not take into account these solvent mol\u00adecules.In the title compound, [{[(C The characterization of the crystals obtained by X-ray diffraction, NMR and IR spectroscopy is in accordance with the formation of the title compound, [{((C6H5)3P)Ag}4{NCO}4], (I)To get access to a large range of metal allophanates 3 moieties. However, the respective components of both disordered Ag(PPh3) units share one phenyl ring (C41\u2013C46 and C59\u2013C64). The Ag4N4-heterocubane is distorted which is reflected by the variation of the Ag\u2014N distances in the range 2.273\u2005(13)\u20132.605\u2005(12)\u2005\u00c5. Likewise, the Ag\u2014N\u2014Ag [78.7\u2005(3) \u2013 98.5\u2005(3)\u00b0] and N\u2014Ag\u2014N [80.9\u2005(3) \u2013 98.5\u2005(3)\u00b0] angles significantly deviate from 90\u00b0. The Ag2N2-faces of the Ag4N4-core are not planar [r.m.s. deviations in the range 0.0293 to 0.1947\u2005\u00c5 ], however, the opposing least-squares planes are almost parallel [angles between planes: 0.40\u2005(3) and 3.2\u2005(3)\u00b0]. Opposing planes are twisted by some degrees relative to each other, which is reflected by the Ag\u2014N\u2014Ag\u2014N and N\u2014Ag\u2014N\u2014Ag torsion angles ranging from 2.8\u2005(3)\u201319.4\u2005(3)\u00b0. As a result of the distortion of the Ag4N4-core, the Ag\u22efAg and N\u22efN separations differ significantly. The shortest distances are observed between Ag1 and Ag2 as well as Ag3/Ag3\u2032 and Ag4/Ag4\u2032 (Table\u00a01I(PPh3) fragment. The scattering contributions of two severely disordered THF solvent mol\u00adecules were treated with the SQUEEZE procedure in PLATON \u00b70.25THF. This discrepancy may be due to a facile evaporation of the co-crystallized solvent under reduced pressure.The title compound consists of a Agde Fig.\u00a01. Each Ag\u2032 Table\u00a01. Conside\u2032 Table\u00a01 a weak a\u2032 Table\u00a01. The Ag\u2014Five moderate-to-weak C\u2014H\u22efO hydrogen bonds in the CSD database \u03bc3-\u03baN coordination of the cyanate anions towards AgI has been described for five compounds only in anhydrous ethanol (40\u2005ml), was added at 273\u2005K ethyl allophanate . The reaction was heated to reflux overnight. The colourless precipitate formed was filtered off, washed thrice with ethanol (each 20\u2005ml) and dried under vacuum (yield: 850\u2005mg). The resulting solid material (407\u2005mg) was dissolved in water (20\u2005ml) and was added dropwise to a solution of silver nitrate in water (15\u2005ml). The suspension obtained was stirred at ambient temperature overnight. Filtration afforded a colourless solid, which was washed with cold water (20\u2005ml) and dried under vacuum (yield: 250\u2005mg). A suspension of this solid (120\u2005mg) in anhydrous THF (20\u2005ml) was treated with PPh3 at 273\u2005K. After stirring overnight at this temperature, the reaction mixture was filtered through a pad of celite. Removal of all volatiles under reduced pressure afforded a pale purple solid . Colourless crystals of (I)To a solution of sodium ethano\u00adlate in ethanol, generated 1H NMR : \u03b4 = 7.40\u20137.28 . 13C{1H} NMR : \u03b4 = 134.0 , 132.1 , 130.4 , 129.1 . The resonance signal of the cyanate carbon atom is not observed under the measurement conditions applied. 31P{1H} NMR : \u03b4 = 9.0 (s). IR : \u03bd = 3449 (w), 3356 (w), 2170 , 1603 (w), 1429 (w), 1388 (w), 1300 (m), 1206 (m), 638 (m).M.p. 458\u2005K (decomp.). viz. (240), (Uiso(H) = 1.2Ueq(C) and a C\u2014H distance of 0.93\u2005\u00c5. Atoms Ag3 and Ag4 and two of the four P atoms of the PPh3 moieties with attached phenyl rings are disordered over two sets of sites, with occupancy ratios of 0.54\u2005(4):0.46\u2005(4) and 0.55\u2005(2):0.45\u2005(2), respectively. A phenyl ring of another PPh3 moiety is likewise disordered over two sets of sites in a 0.67\u2005(5):0.33\u2005(5) ratio. The disordered phenyl rings were treated by rigid-group refinements. If necessary, the respective C\u2014P distances were restrained to 1.85\u2005(2)\u2005\u00c5. Anisotropic displacement parameters of all atoms were restrained using enhanced rigid-bond restraints I, New_Global_Publ_Block. DOI: 10.1107/S2056989015017636/wm5201Isup2.hklStructure factors: contains datablock(s) I. DOI: 1426238CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Fdd2 space group. Instead, there are two mol\u00adecules in the asymmetric unit, and both of them display a pseudo-trans conformation.The title compound was obtained in crystalline form after preparative HPLC. Conformation of the proposed mol\u00adecular structure was obtained by single-crystal X-ray analysis at 173\u2005K. The mol\u00adecules do not take advantage of the twofold axis provided as an available symmetry option by the 14H16N2S3, crystallized with two independent mol\u00adecules [(1a) and (1b)] in the asymmetric unit. Both mol\u00adecules display a pseudo-trans conformation. The two consecutive S\u2014S bond lengths of the tris\u00adulfane unit of mol\u00adecule (1a) are 2.06\u2005(3) and 2.08\u2005(3)\u2005\u00c5, and 2.08\u2005(3) and 2.07\u2005(2)\u2005\u00c5 for mol\u00adecule (1b). Torsion angles about each of the two S\u2014S bonds are 86.6\u2005(2) and 87.0\u2005(2)\u00b0 for (1a), and \u221284.6\u2005(2) and \u221285.9\u2005(2)\u00b0 for (1b). The core atoms, viz. the N\u2014S\u2014S\u2014S\u2014N moiety, of the two mol\u00adecules superimpose well if one is inverted on the other, but the phenyl groups do not. Thus, the two units are essentially conformational enanti\u00adomers. In mol\u00adecule (1a), the two phenyl rings are inclined to one another by 86.7\u2005(3)\u00b0, and in mol\u00adecule (1b), by 81.1\u2005(3)\u00b0. In the crystal, mol\u00adecules are linked via C\u2014H\u22ef\u03c0 inter\u00adactions, forming sheets lying parallel to (010).The title compound, C The mol\u00adecules do not take advantage of the twofold axis provided as an available symmetry option by the Fdd2 space group. Instead, there are two mol\u00adecules, (1a) and (1b), in the asymmetric unit . All bond distances and angles in both mol\u00adecules are within expected ranges. Selected geometric parameters for compound (1) are given in Table\u00a011a) are 2.064\u2005(3) and 2.078\u2005(3)\u2005\u00c5, and for mol\u00adecule (1b) are 2.076\u2005(3) and 2.067\u2005(2)\u2005\u00c5. These values are similar to the value of 2.07\u2005\u00c5 reported for the S\u2014S bond length in elemental sulfur (S8). Torsion angles about each of the two S\u2014S bonds (comprising the tris\u00adulfane) are, respectively, 86.6\u2005(2) and 87.0\u2005(2)\u00b0 for (1a), and \u221284.6\u2005(2) and \u221285.9\u2005(2)\u00b0 for (1b). The core atoms, viz. the N\u2014S\u2014S\u2014S\u2014N moiety, of the two units superimpose well if one is inverted on the other, but the phenyl groups do not. Thus, the two units are essentially conformational enanti\u00adomers. Moreover, with respect to the four measured torsion angles, which range in absolute value from 84.6\u2005(2) to 87.0\u2005(2)\u00b0, these are slightly smaller than the theoretical optimum of 90.0\u00b0 . The theoretically possible pseudo-cis conformation , mol\u00adecules are linked via C\u2014H\u22ef\u03c0 inter\u00adactions, forming sheets lying parallel to (010) \u00adtris\u00adulfane, (2) amino]\u00adtris\u00adulfane, (3) , compounds (2) and (3) each have a unique conformation in the unit cell (Z\u2032 = 1). Selected geometric parameters of (1) and the comparison compounds, (2) and (3), are given in Table\u00a01ca 2.07\u2005\u00c5, the corresponding value is longer (2.09\u2005\u00c5) in (3) and shorter (2.04\u2005\u00c5) in (2). The absolute value of the average torsion angle of the title compound (1) is ca 86.0\u00b0, while the corresponding value is larger (93.2 and \u221289.5\u00b0) and closer to the theoretical optimum in (2), and significantly larger (109.7 and 95.9\u00b0) in (3).A search of the Cambridge Structural Database \u00adtris\u00adulfan, (2) amino]\u00adtris\u00adulfan, (3) , was synthesized and obtained in crystalline form after preparative HPLC, as described by Schroll & Barany (198637) in that publication.The title compound, (rany 1986: compounUiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015011342/su5144sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989015011342/su5144Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015011342/su5144Isup3.cmlSupporting information file. DOI: 1406065CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystals of both compounds, mol\u00adecules are linked 13H8ClN3O6, (I), and C13H9N3O6, (II), differ in the orientation of the two aromatic rings. In (I), they are essentially coplanar, making a dihedral angle of 8.2\u2005(1)\u00b0, while in (II), they are inclined to one another by 76.2\u2005(1)\u00b0. The two nitro groups are essentially coplanar with the attached benzene rings, as indicated by the dihedral angles of 1.4\u2005(2) and 2.3\u2005(2)\u00b0 in (I), and 4.96\u2005(18) and 5.4\u2005(2)\u00b0 in (II). The carbamate group is twisted slightly from the attached benzene ring, with a C\u2014N\u2014C\u2014O torsion angle of \u2212170.17\u2005(15)\u00b0 for (I) and 168.91\u2005(13)\u00b0 for (II). In the crystals of of both compounds, mol\u00adecules are linked via N\u2014H\u22efO hydrogen bonds, forming chains propagating along [010]. In (I), C\u2014H\u22efO hydrogen bonds also link mol\u00adecules within the chains. The crystal packing in (I) also features a very weak \u03c0\u2013\u03c0 inter\u00adaction [centroid\u2013centroid distance = 3.7519\u2005(9)\u2005\u00c5].The title compounds, C Within via N\u2014H\u22efO hydrogen bonds, this time involving the carbonyl O atom O5, forming chains propagating along the b axis; see Table\u00a02In the crystal of (II)N-phenyl\u00adcarbamate gave 16 hits for similar compounds, including two ortho\u00adrhom\u00adbic poylmorphs of phenyl N-phenyl\u00adcarbamate itself \u00b0 in (I)A search of the Cambridge Structural Database dissolved in 100\u2005ml of dry THF, and to it was added the calculated amount (with 5% excess) of 4-chloro\u00adphenyl\u00adchloro\u00adformate for compound (I)Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015010245/su5141sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989015010245/su5141Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989015010245/su5141IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989015010245/su5141Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015010245/su5141IIsup5.cmlSupporting information file. DOI: 1403525, 1403524CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal of the former, mol\u00adecules are linked 25H29BrN5O7, (I) dioxola-1-benzena\u00adcyclo\u00adnona\u00adphane], and C24H29N5O7S, (II) thia\u00adzola-6-triazola-3-furodioxola-1-benzena\u00adcyclo\u00adnona\u00adphane], the triazole rings adopt almost planar conformations. In (I), the fused pyrrolidine rings adopt envelope conformations with the C atoms opposite the fused N\u2014C bond as the flap in each ring, and their mean planes are inclined to one another by 52.8\u2005(3)\u00b0. In (II), the pyrrolidine and thia\u00adzole rings are both twisted on the fused N\u2014C bond, and their mean planes are inclined to one another by 70.8\u2005(2)\u00b0. In both (I) and (II), the furan ring adopts an envelope conformation with the adjacent C atom of the macrocycle as the flap. In the crystal of (I), mol\u00adecules are linked via C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds, forming sheets parallel to (10-1), while in (II), mol\u00adecules are linked via C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds, forming helical chains propagating along [010], which are linked via C\u2014H\u22efS hydrogen bonds, forming slabs parallel to (001).In the title compounds, C In compound (I)D = N1/C11\u2013C13/C7 and E = N1/C8\u2013C11) and the furan ring (B = O3/C15/C19/C20/C14) have envelope conformations with the maximum deviations from the respective mean planes of 0.465\u2005(5)\u2005\u00c5 for atom C13, 0.490\u2005(7)\u2005\u00c5 for C9 and 0.500\u2005(4)\u2005\u00c5 for C14. The dioxalane ring (C = O4/C15/C19/O5/C16) has a twisted conformation on bond O5\u2014C15. The mean planes of rings B and C are inclined to one another by 70.0\u2005(3)\u00b0, and the mean planes of rings D and E are inclined to one another by 52.8\u2005(3)\u00b0. The molecular structures of compounds (I) and (II) are illustrated in Figs. 1D) and thia\u00adzole rings (E = N1/C8/S9/C10/C11) have twist conformations on bond N1\u2014C11. The furan and dioxolane rings (B and C) adopt envelope conformations with maximum deviations from the mean planes of 0.631\u2005(3)\u2005\u00c5 for atom C14 and 0.319\u2005(4)\u2005\u00c5 for C16. The mean planes of rings B and C are inclined to one another by 68.5\u2005(2)\u00b0 and the mean planes of rings D and E are inclined to one another by 70.8\u2005(2)\u00b0. This latter dihedral angle is much larger than that in compound (I)cf. 52.8\u2005(3)\u00b0.In compound (II)A) makes dihedral angles of 74.0\u2005(3), 65.8\u2005(3) and 65.8\u2005(3)\u00b0 with the mean planes of rings B and D and the benzene ring (C1\u2013C6), respectively. The corresponding dihedral angles in compound (II)A/B and A/D; 74.0\u2005(3) and 65.8\u2005(3), respectively, for (IIn compound (I)via C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds, forming sheets parallel to ; Table\u00a02In the crystal of (I)Compound (I): A solution of 5-bromo-2--2,2-dimethyl-5-[(Z)-2-nitro\u00advin\u00adyl]tetra\u00adhydro\u00adfurodi\u00adoxol-6-yl}\u00adoxy)meth\u00adyl]-1H-1,2,3-triazol-1-yl}eth\u00adoxy)benzalde\u00adhyde (1\u2005mmol) and proline (1.5\u2005mmol) was refluxed in dry aceto\u00adnitrile (50\u2005ml) under a nitro\u00adgen atmosphere for 9\u2005h. After completion of the reaction, as indicated by TLC, the aceto\u00adnitrile was evaporated under reduced pressure. The crude product was purified by column chromatography using hexa\u00adne/EtOAc (3:7) as eluent (yield 75%). After purification the compound was recrystallized in CHCl3 by slow evaporation yielding colourless block-like crystals.Compound (II): A solution of 5-bromo-2--2,2-dimethyl-5-[(Z)-2-nitro\u00advin\u00adyl]tetra\u00adhydro\u00adfurodioxol-6-yl}\u00adoxy)meth\u00adyl]-1H-1,2,3-triazol-1-yl}eth\u00adoxy)benzalde\u00adhyde (1\u2005mmol) and thia\u00adzolidine-4-carb\u00adoxy\u00adlic acid (1.5\u2005m\u2005mol) was refluxed in dry aceto\u00adnitrile (50\u2005ml) under a nitro\u00adgen atmosphere for 9\u2005h. After completion of reaction, as indicated by TLC,the aceto\u00adnitrile was evaporated under reduced pressure. The crude product was purified by column chromatography using hexa\u00adne/EtOAc (4:6) as eluent (yield 75%). After purification the compound was recrystallized in CHCl3 by slow evaporation yielding colourless block-like crystals.Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms. Compound (I) was refined using the instructions TWIN/BASF global, I, II. DOI: 10.1107/S205698901501141X/su5103Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S205698901501141X/su5103IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1023614, 1023839CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atom is located on an inversion centre and is coordinated by two 4-cyano\u00adbenzoate (CNB) anions, two nicotinamide (NA) ligands and two water mol\u00adecules. The four O atoms in the equatorial plane form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination sphere is completed by the two N atoms of the NA ligands in the axial positions.In the title complex, the Co 8H4NO2)2(C6H6N2O)2(H2O)2], the CoII atom is located on an inversion centre and is coordinated by two 4-cyano\u00adbenzoate (CNB) anions, two nicotinamide (NA) ligands and two water mol\u00adecules. The four O atoms in the equatorial plane form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination sphere is completed by the two N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 22.11\u2005(15)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 89.98\u2005(5)\u00b0. In the crystal, inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds link the mol\u00adecules, enclosing R22(8) and R44(8) ring motifs, forming layers parallel to (100). The layers are linked via C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds, resulting in a three-dimensional network. A weak C\u2014H\u22ef\u03c0 inter\u00adaction is also observed.In the title complex, [Co(C N,N-di\u00adethyl\u00adnicotinamide (DENA) is an important respiratory stimulant is one form of niacin. A deficiency of this vitamin leads to loss of copper from the body, known as pellagra disease. Victims of pellagra show unusually high serum and urinary copper levels bis\u00ad(nico\u00adtinamide-\u03baN1)cobalt(II), [Co(C8H4O2N)2(C6H6N2O)2(H2O)2], and report herein its crystal structure.The structure\u2013function\u2013coordination relationships of the aryl\u00adcarboxyl\u00adate ion in ZnII atom is located on an inversion centre and is coordinated by two 4-cyano\u00adbenzoate (CNB) anions, two nicotinamide (NA) ligands and two water mol\u00adecules, with all ligands coordinating in a monodentate manner and the two symmetry-related water O atoms (O4 and O4i) form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination sphere is completed by the two symmetry-related N atoms (N2 and N2i) of the two NA ligands in the axial positions \u2005\u00c5] and C1\u2014O2 [1.256\u2005(2)\u2005\u00c5], bond lengths of the carboxyl\u00adate group indicate delocalized bonding arrangements, rather than localized single and double bonds. The Co\u2014O bond lengths are 2.0835\u2005(12)\u2005\u00c5 (for benzoate oxygen atoms) and 2.1350\u2005(13)\u2005\u00c5 (for water oxygen atoms), and the Co\u2014N bond length is 2.1390\u2005(15)\u2005\u00c5, close to standard values. The Co1 atom lies 0.3921\u2005(1)\u2005\u00c5 above the planar (O1/O2/C1) carboxyl\u00adate group. The O\u2014Co\u2014O and O\u2014Co\u2014N bond angles deviate only slightly from ideal values, with average values of 90\u2005(3)\u00b0 and 90\u2005(2)\u00b0, respectively.A (C2\u2013C7)] is 22.11\u2005(15)\u00b0, while the benzene and pyridine [B (N2/C9\u2013C13)] rings are oriented at a dihedral angle of 89.98\u2005(5)\u00b0.The dihedral angle between the planar carboxyl\u00adate group (O1/O2/C1) and the adjacent benzene ring [c (c = carboxyl\u00adate), N\u2014H\u22efOn (n = nicotinamide), O\u2014Hw\u22efOc (w = water) and O\u2014Hw\u22efOn hydrogen bonds and C\u2014Hn\u22efNcnb hydrogen bonds in H2O (50\u2005ml) and nicotinamide in H2O (50\u2005ml) with sodium 4-cyano\u00adbenzoate in H2O (100\u2005ml). The mixture was filtered and set aside to crystallize at ambient temperature for several days, giving pink-coloured single crystals.The title compound was prepared by the reaction of CoSO2) and H41 and H42 (for H2O) were located in a difference Fourier map and were refined freely. The aromatic C-bound H atoms were positioned geometrically with C\u2014H = 0.93\u2005\u00c5, and constrained to ride on their parent atoms, with Uiso(H) = 1.2Ueq(C). The highest electron density and the deepest hole were found 0.80\u2005\u00c5 and 0.83\u2005\u00c5, respectively, from Co1.The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0210.1107/S2056989015008270/wm5151sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015008270/wm5151Isup2.hklStructure factors: contains datablock(s) I. DOI: 1061935CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III ion in the title compound shows a slightly distorted FeCl2N2O2 octa\u00adhedral coordination geometry. In the crystal, two complex mol\u00adecules are linked by duplex O\u2014H\u22efO hydrogen bonds. Additional hydrogen-bonding inter\u00adactions lead to the formation of undulating sheets parallel to (010).The Fe 10H15N2O2)Cl2]\u00b72H2O, the FeIII ion is coordinated by two N and two O atoms of the tetra\u00addentate 2-{(2-hy\u00addroxy\u00adeth\u00adyl)(pyridin-2-ylmeth\u00adyl)amino}\u00adethano\u00adlate ligand and by two chloride anions, resulting in a distorted octa\u00adhedral coordination sphere. The average Fe\u2014X (X = ligand N and O atoms) and Fe\u2014Cl bond lengths are 2.10 and 2.32\u2005\u00c5, respectively. In the crystal, duplex O\u2014H\u22efO hydrogen bonds between the hydroxyl and eth\u00adoxy groups of two neighbouring complexes give rise to a dimeric unit. The dimers are connected to the lattice water mol\u00adecules through O\u2014H\u22efCl hydrogen bonds, forming undulating sheets parallel to (010). Weak C\u2014H\u22efCl hydrogen bonds are also observed.In the title compound, [Fe(C The chloride anions are trans to the deprotonated eth\u00adoxy O atom and the N atom of the pyridine group of the Hpmide ligand, respectively, and coordinate in cis position to each other. The average Fe\u2014XHpmide bond length is 2.10\u2005\u00c5 and the Fe\u2014Cl bond lengths are 2.2773\u2005(5) and 2.3581\u2005(7) \u00c5. Both the average Fe\u2014N (2.182\u2005\u00c5) and Fe\u2014O (2.010\u2005\u00c5) distances in (I)2O2-chelated high-spin FeIII complexes The hydroxyl substituent of the Hpmide ligand forms a strong hydrogen bond with the O atom of the deprotonated eth\u00adoxy group of a neighbouring mol\u00adecule. These duplex inter\u00adactions lead to a dinuclear dimeric unit. The dimers are linked through O\u2014H\u22efCl inter\u00adactions to the lattice water mol\u00adecules, that are likewise connected to each other through O\u2014H\u22efO hydrogen bonds. All these hydrogen-bonding inter\u00adactions was added dropwise a MeOH solution (3\u2005ml) of H2pmide . The colour became yellow, and then the solution was stirred for 30\u2005min at room temperature. Yellow crystals of (I)10H15Cl2FeN2O2: C 37.30, H 4.70, N 8.70%; found: C 37.19, H 4.58, N 8.78%.The HUiso(H) values of 1.2Ueq(C) of the parent atoms. One lattice water mol\u00adecule (OW1) was found to be equally disordered over two positions. The H atoms of this disordered water mol\u00adecule (H1W1 and H1W2) were located from difference Fourier maps and refined with restraints and a fixed O\u2014H distances of 0.84\u2005\u00c5, with Uiso(H) values of 1.2Ueq(O). Moreover, the second water mol\u00adecule (O2W) was modelled without hydrogen atoms because difference Fourier maps did not suggest suitable H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814022089/wm5071sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814022089/wm5071Isup2.hklStructure factors: contains datablock(s) I. DOI: 1027864CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Supra\u00admolecular chains about a 4 10H7N3O4S, the dihedral angle between the terminal five-membered rings is 27.4\u2005(2)\u00b0, with these lying to the same side of the plane through the central CN2C(=O) atoms (r.m.s. deviation = 0.0403\u2005\u00c5), leading to a curved mol\u00adecule. The conformation about the C=N imine bond [1.281\u2005(5)\u2005\u00c5] is E, and the carbonyl O and amide H atoms are anti. In the crystal, N\u2014H\u22efO hydrogen bonds lead to supra\u00admolecular chains, generated by a 41 screw-axis along the c direction. A three-dimensional architecture is consolidated by thienyl-C\u2014H\u22efO(nitro) and furanyl-C\u2014H\u22efO(nitro) inter\u00adactions, as well as \u03c0\u2013\u03c0 inter\u00adactions between the thienyl and furanyl rings [inter-centroid distance = 3.515\u2005(2)\u2005\u00c5]. These, and other, weak inter\u00admolecular inter\u00adactions, e.g. nitro-N\u2014O\u22ef\u03c0(thien\u00adyl), have been investigated by Hirshfeld surface analysis, which confirms the dominance of the conventional N\u2014H\u22efO hydrogen bonding to the overall mol\u00adecular packing.In the title carbohydrazide, C The thienyl ring is also planar within experimental error [r.m.s. deviation = 0.005\u2005\u00c5] and orientated so that the sulfur atom is syn to the carbonyl-O1 atom. Overall, the mol\u00adecule is curved with the rings lying to the same side of the plane through the bridging CN2C(=O) atoms, r.m.s. deviation = 0.0403\u2005\u00c5, with twists noted in both the S1\u2014C1\u2014C5\u2014O1 and N2\u2014C6\u2014C7\u2014O2 torsion angles of \u22129.8\u2005(5) and 5.4\u2005(6)\u00b0, respectively; the dihedral angle between the five-membered rings is 27.4\u2005(2)\u00b0.In (I)anti relationship between the carbonyl-O and amide-H atoms enables the formation of directional N\u2014H\u22efO hydrogen bonds leading to supra\u00admolecular chains, generated by a 41 screw-axis propagating along the c-axis direction, Fig.\u00a02a and Table\u00a01y, x, \u2212z. A view of the unit-cell contents is shown in Fig.\u00a02b.The Crystal Explorer 3.1 Cg = 3.506\u2005(4)\u2005\u00c5, O4\u22efCg = 3.639\u2005(4)\u2005\u00c5 and N3\u2014O4\u22efCg = 74.0\u2005(2)\u00b0] is also evident from the light-blue and red regions corresponding to their respective potentials on the Hirshfeld surface mapped over electrostatic potential in Fig.\u00a04The presence of a short inter\u00admolecular C\u22efC contact between thienyl-C2 and furanyl-C10 atoms, Table\u00a02a and those delineated into O\u22efH/H\u22efO, H\u22efH, N\u22efH/H\u22efN, C\u22efH/H\u22efC, C\u22efC, C\u22efO/O\u22efC and S\u22efH/H\u22efS contacts distances greater than their van der Waals separations with the comparatively low contribution, i.e. 13.6%, due to the relatively low hydrogen-atom content in the mol\u00adecule. The absence of characteristic wings in the fingerprint plot delineated into C\u22efH/H\u22efC and the low contribution to the Hirshfeld surface, Fig.\u00a06e and Table\u00a03de + di \u223c2.9\u2005\u00c5 belong to short inter\u00adatomic C\u22efH contacts, Table\u00a02de + di \u223c2.7\u2005\u00c5 in the fingerprint plot corresponding to N\u22efH/H\u22efN contacts, Fig.\u00a06e, with a 7.5% contribution to the Hirshfeld surface is the result of short inter\u00adatomic N\u22efH/H\u22efN contacts, Table\u00a02de = di \u223c1.7\u2005\u00c5, Fig.\u00a06f. The presence of \u03c0\u2013\u03c0 stacking inter\u00adactions between the symmetry-related thienyl and furanyl rings is also indicated by the appearance of red and blue triangle pairs on the Hirshfeld surface mapped with the shape-index property identified with arrows in the images of Fig.\u00a07h, they do not have a significant influence on the mol\u00adecular packing as they are separated at distances greater than the sum of their van der Waals radii.The overall two-dimensional fingerprint plot is shown in Fig.\u00a06et al., 2014i.e. 0.99, corresponding to the C\u22efO/O\u22efC contacts. The high propensity to form \u03c0\u2013\u03c0 stacking inter\u00adactions between the thienyl and furanyl rings is reflected from the high enrichment ratio 2.66 for C\u22efC contacts. The ER value of 1.26 resulting from 6.75% of the surface occupied by nitro\u00adgen atoms and a 7.5% contribution to the Hirshfeld surface from N\u22efH/H\u22efN contacts is due to the presence of short N\u22efH contacts in the structure, Table\u00a02The final analysis based on the Hirshfeld surfaces is an evaluation of enrichment ratios (ER) methyl\u00adene]thio\u00adphene-2-carbohydrazide +; = 1.2Ueq(C). The N-bound H atom was located from a difference map and refined with (N\u2014H = 0.88\u00b10.01\u2005\u00c5), and with Uiso(H) = 1.2Ueq(C). The slightly elongated displacement ellipsoid for the C2 atom in the thienyl ring is likely due to unresolved disorder in the ring where the second, co-planar orientation related by 180\u00b0 to that modelled is present. However, this was not modelled as the maximum residual electron density peak was only 0.46\u2005e\u2005\u00c5\u22123, 0.61\u2005\u00c5 from the C2 atom. It is also noted that the relevant S\u2014C and C\u2014C bond lengths show the expected values.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989016009968/hb7593sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016009968/hb7593Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016009968/hb7593Isup3.cmlSupporting information file. DOI: 1486459CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure consists of hydrogen-bonded planar arrays held by X\u2014H\u22efO and X\u2014O\u22ef\u03c0(ring) inter\u00adactions, leaving inter\u00adstitial columnar voids.A dinuclear Cd 3-carb\u00adoxy-6-methyl\u00adpyridine-2-carboxyl\u00adate complex is reported, in which the anionic ligand displays an unusual \u03bc 3N,O2:O2;\u03ba3O2:N,O2-bis\u00ad[(nitrato-\u03baO)cadmium] methanol monosolvate, [Cd2(C8H6NO4)2(NO3)2(C10H8N2)2]\u00b7CH3OH, was isolated as colourless crystals from the reaction of Cd(NO3)2\u00b74H2O, 6-methyl\u00adpyridine-2,3-di\u00adcarb\u00adoxy\u00adlic acid (mepydcH2) and 2,2\u2032-bi\u00adpyridine in methanol. The asymmetric unit consists of a CdII cation bound to a \u03bc-\u03ba3N,O2:O2-mepydcH\u2212 anion, an N,N\u2032-bidentate 2,2\u2032-bi\u00adpyridine group and an O-mono\u00addentate nitrate anion, and is completed with a methanol solvent mol\u00adecule at half-occupancy. The Cd complex unit is linked to its centrosymmetric image through a bridging mepydcH\u2212 carboxyl\u00adate O atom to complete the dinuclear complex mol\u00adecule. Despite a significant variation in the coordination angles, indicating a considerable departure from octa\u00adhedral coordination geometry about the CdII atom, the Cd\u2014O and Cd\u2014N distances in this complex are surprisingly similar. The crystal structure consists of O\u2014H\u22efO hydrogen-bonded chains parallel to a, further bound by C\u2014H\u22efO contacts along b to form planar two-dimensional arrays parallel to (001). The juxtaposed planes form inter\u00adstitial columnar voids that are filled by the methanol solvent mol\u00adecules. These in turn inter\u00adact with the complex mol\u00adecules to further stabilize the structure. A search in the literature showed that complexes with the mepydcH\u2212 ligand are rare and complexes reported previously with this ligand do not adopt the \u03bc-\u03ba3 coordination mode found in the title compound.The centrosymmetric dinuclear complex bis\u00ad(\u03bc-3-carb\u00adoxy-6-methyl\u00adpyridine-2-carboxyl\u00adato)-\u03ba The two deprotonated forms pydcH\u2212 and 2\u2212pydc have been shown to adopt a wide range of coordination modes through their carboxyl\u00adate oxygen and pyridyl nitro\u00adgen atoms , only a single structure was found involving the monoanionic mepydcH\u2212 ligand similar to that reported here complexes [M(bpee)2(mepydcH)2] ethyl\u00adene) with octa\u00adhedral geometry around MII. Both mepydcH\u2212 fragments act in a simple \u03ba22N,O-chelating mode binding to a single nucleus while the two N-bound bpee ligands are trans-monodentate. The formation of these mononuclear complexes is unusual considering the obvious bridging potential of the bpee ligands. Mixed-ligand complexes based on non-methyl\u00adated 2,3-pyridinedi\u00adcarboxyl\u00adate and 4,4\u2032-bi\u00adpyridine-like ligands usually generate stable polymeric structures with the exo-bidentate ligands adopting a bridging role 2(mepydcH)2(NO3)2]\u00b7MeOH (I)2- coordination behaviour and the fact that the ligand is only singly deprotonated has no counterpart in complexes of the non-methyl\u00adated ligands and makes this a genuinely novel structure. The closest relatives with 2,2\u2032-bi\u00adpyridine as the auxiliary ligand are found with di-anionic 2\u2212pydc ligands, but these are either mononuclear chelates through the pyridine N and carboxyl\u00adate O atoms. A chelating 2,2\u2032-bi\u00adpyridine that binds through both nitro\u00adgen atoms and a unidentate nitrate anion complete the coordination sphere; the asymmetric unit also contains a non-coordinating half-occupancy methanol solvate. This five coordinate CdII unit, in turn, binds to its centrosymmetric image through the carboxyl\u00adate oxygen atom of the mepydcH\u2212 ligand, forming a pair of Cd\u2013O\u2013Cd bridges. As a result, a dimeric unit forms distances are reasonable, spanning the range 2.304\u2005(2)\u20132.332\u2005(3)\u2005\u00c5. However, the coordination angles vary widely ; the result is a rather distorted octa\u00adhedral geometry around Cd1. Selected geometric parameters are shown in Table\u00a01The complex consists of a Cdms Fig.\u00a01 with eacviz., hydrogen bonds Contacts #1 when a lone pair provided by a carbonyl oxygen points toward the centroid of an aromatic ring Strong inter\u00admolecular O\u2014H\u22efO contacts #2 C\u2014H\u22efO inter\u00adactions #3, #4 and #5 . These planes juxtapose along [001] with rather weak direct inter\u00adactions. In the process, however, significant columnar voids parallel to the chains are formed in which the partial occupancy methanol solvate mol\u00adecules reside. These are not free, but enter instead into a number of weak C\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22ef\u03c0 interactions Fig.\u00a03a. ThesemepydcH2, 2.2\u2032-bi\u00adpyridine and (I)Resolutions Pro software, using a Diamond ATR accessory. The FT\u2013IR spectrum of (I)\u22121 range, and confirms the structural data indicating the presence of the coordinating nitrate and mepydcH\u2212 anions. Bands due to the unidentate NO3\u2212 group were found at 1478 and 1298\u2005cm\u22121 and appear due to the \u03bdasym(ONO) and \u03bdsym(ONO) vibrations, with a shoulder at 1010\u2005cm\u22121 due to the \u03bd(NO) stretching modes of nitrate groups \u22121, \u03bd(OH) stretching for a hydrogen-bonded system stretch. The deprotonated carboxyl\u00adate (COO\u2212) is characterized by the asymmetric and symmetric stretching modes \u03bdas at 1593\u2005cm\u22121 and \u03bds at 1322\u2005cm\u22121. This confirms the unidentate coordination of the carboxyl\u00adate O atom, with the difference between these frequencies being > 200\u2005cm\u22121(\u0394 = \u03bdas \u2212 \u03bds = 271\u2005cm\u22121) of almost equal intensity due to the \u03bd(C=C) + \u03bd(C=N) vibrations from the coordinating 2,2\u2032-bi\u00adpyridine ligand \u00b74H2O and mepydcH2 in MeOH (4.0\u2005mL). The mixture was stirred to dissolve the 2,2\u2032-bi\u00adpyridine and was then allowed to stand undisturbed at room temperature in an uncovered 10\u2005mL beaker. Colourless single crystals of compound (I)Solid 2,2\u2032-bi\u00adpyridine was added to a solution prepared by disolving Cd = 1.2Ueq(C); C\u2014Hmeth\u00adyl: 0.97\u2005\u00c5, Ueq(H) = 1.5Ueq(C). The O\u2014H hydrogen atom was refined with a restrained O\u2014H distance [0.85\u2005(1)\u00c5], and with U(H) = 1.2Ueq(O). The methanol solvate was refined at half occupancy.Relevant crystallographic data for (I)10.1107/S2056989015012384/sj5468sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015012384/sj5468Isup2.hklStructure factors: contains datablock(s) I. DOI: 1409269CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound is an example of a \u2018false conglomerate\u2019 with two mol\u00adecules of opposite handedness in the asymmetric unit. Each mol\u00adecule exists as a zwitterion, with proton transfer from the amino acid carb\u00adoxy\u00adlic acid group to the amine group. In the crystal, the components are linked by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, generating (100) sheets. 6H9NO4\u00b7H2O [systematic name: -rel-\u03b1-amino-2-carb\u00adoxy\u00adcyclo\u00adpropane\u00adacetic acid monohydrate], crystallizes with two organic mol\u00adecules and two water mol\u00adecules in the asymmetric unit. The space group is P21 and the organic mol\u00adecules are enanti\u00adomers, thus this is an example of a \u2018false conglomerate\u2019 with two mol\u00adecules of opposite handedness in the asymmetric unit (r.m.s. overlay fit = 0.056\u2005\u00c5 for one mol\u00adecule and its inverted partner). Each mol\u00adecule exists as a zwitterion, with proton transfer from the amino acid carb\u00adoxy\u00adlic acid group to the amine group. In the crystal, the components are linked by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, generating (100) sheets. Conformationally restricted glutamate analogs are of inter\u00adest due to their selective activation of different glutamate receptors, and the naturally occurring (+)-CCG-III is an inhibitor of glutamate uptake and the key geometrical parameters are discussed.The title compound, C CCG-I, CCG-III and CCG-IV glycines IV Fig.\u00a01 are natuII Fig.\u00a01 have beeThe racemic title compound Fig.\u00a02 crystallX = \u22124.3\u00b0 and O3A\u2014C6A\u2014C2A\u2014X = \u221211.1\u00b0 indicate that the carb\u00adoxy\u00adlic acid attached to the cyclpropane ring adopts a bis\u00adected conform\u00adation ; C2A\u2014C3A = 1.516\u2005(2)\u2005\u00c5] and are longer than the cyclo\u00adpropane bonds distal to the C2 carb\u00adoxy\u00adlic acid . These distances and torsion angles are consistent with other cyclo\u00adpropane carb\u00adoxy\u00adlic acids , between the \u03b1- and \u03b3-carboxyl\u00adate carbon atoms (2d), and their sum (1d + 2d). The classifications \u2018folded\u2019, \u2018semi-folded\u2019, \u2018semi-extended\u2019, and \u2018extended\u2019 are defined by (1d + d2) \u2264 7.5\u2005\u00c5, 7.5\u2005\u00c5 \u2264 (1d + d2) \u2264 8.0\u2005\u00c5, 8.0\u2005\u00c5 \u2264 (1d + d2) \u2264 8.5\u2005\u00c5, and (1d + d2) \u2265 8.5\u2005\u00c5, respectively , 8.24 and 8.30\u2005\u00c5, respectively. From these values, these conformers of CCG-III can be considered to be in the \u2018semi-extended\u2019 class.Conformationally restricted glutamic acid analogs can be classified into one of four categories, which are characterized by the distances between the nitro\u00adgen atom of the amino group and the \u03b3-carboxyl\u00adate carbon atom ; Table\u00a01The racemic title compound was prepared according to the literature procedure I, New_Global_Publ_Block. DOI: 10.1107/S2056989015011500/hb7407Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015011500/hb7407Isup3.cmlSupporting information file. DOI: 1406594CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III irida\u00adbenzenes that contain an open coordination site.Treatment of an irida\u00adbenzene with either bromine or iodine generates high-oxidation-state Ir 3)2], with either bromine or iodine produced di\u00adbromido\u00ad(tri\u00adphenyl\u00adphosphine)iridium(III), [IrBr2{\u2014C(Ph)=C(Ph)\u2014CH=CH\u2014CH=}(PPh3)], (I), and di\u00adiodido\u00ad(tri\u00adphenyl\u00adphosphane)iridium(III), [IrI2{\u2014C(Ph)=C(Ph)\u2014CH=CH\u2014CH=}(PPh3)], (II), respectively, which are two rare examples of 16-electron metalla\u00adbenzenes. Structural elucidation of (I) and (II) reveals that these isotypic irida\u00adbenzenes are unusual, not only in their electron count, but also in their coordination sphere of the IrIII atom where they contain an apparent open coordination site. The crystal structures of (I) and (II) confirm that the mol\u00adecules are complexes containing five-coordinated IrIII with only one tri\u00adphenyl\u00adphosphine group bound to the iridium atom, unambiguously proving that the mol\u00adecules are indeed 16-electron, high-oxidation-state irida\u00adbenzenes. The coordination geometry of the IrIII atom in both structures can be best described as a distorted square pyramid with the P, two Br (or I) and one C atom in the basal plane and another C atom in the apical position.Treatment of carbon\u00adylbis\u00ad(tri\u00adphenyl\u00adphosphane)iridium, [IrCO(\u2014C(Ph)=C(Ph)\u2014CH=CH\u2014CH=)(PPh The mol\u00adecular structures of (I)III is five-coordinated in these complexes with only one tri\u00adphenyl\u00adphosphine group bound to the iridium atom, unambiguously proving that the mol\u00adecules are indeed 16-electron, high-oxidation-state irida\u00adbenzenes. The coordination geometry of the IrIII atom in both structures can be best described as a distorted square pyramid with the P1, Br1(I1), Br2(I2) and C1 atoms in the basal plane and the C5 atom in the apical position. The Br1(I1), Br2(I2), P1, C1 fragments are planar within 0.17\u2005\u00c5 (Br) and 0.21\u2005\u00c5 (I)I irida\u00adbenzenes (2.01\u20132.05\u2005\u00c5), reflecting the higher IrIII oxidation state et al., 2007cf, Br1(I1)\u2014Ir\u2014C5, 110.5\u2005(2) [113.6\u2005(4)\u00b0]}. This site is typically occupied by CO in all of our previous irida\u00adbenzene studies, such as (III) in (I)ca 3.18\u2005\u00c5. This H29A atom is on the opposite side from the C5 atom (the C5\u2014Ir1\u22efH29A angle is 147\u00b0). If present, the Ir\u2014H distance would be around 1.5\u20131.6\u2005\u00c5. In such a case, the distance between this H atom and the H29A atom from the phenyl ring should be 1.6\u20131.7\u2005\u00c5. This distance is too short as a typical H\u22efH contact is 2.4\u2005\u00c5. It follows then that if one H atom does not fit, H2 will not either. The displacement parameters of most C atoms in the phenyl rings are elongated perpendicular to the average plane of the Ph rings showing their flexibility or statistical disorder.Additionally, the irida\u00adbenzene ring in both structures significantly deviates from planarity (Zhu I) Fig.\u00a03. The steX inter\u00adactions with C\u22efX distances in the ranges of 3.533\u2005(7)\u2013 3.717\u2005(5) and 3.699\u2005(17)\u20133.707\u2005(12)\u2005\u00c5, respectively, for Br and I (Tables 1Compounds (I) Tables 1 and 2 \u25b8.et al., 1999Reaction of irida\u00adbenzene (Gilbertson Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015018952/wm5215sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989015018952/wm5215Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989015018952/wm5215IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1430096, 1430095CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III(C48H36N4O2)(H2O)2][K(C12H24O6)(H2O)2](SO3CF3)2\u00b72C12H24O6, the FeIII atom is situated on an inversion centre and is octa\u00adhedrally coordin\u00adated by four pyrrole N atoms of the deprotenated 5,10,15,20-tetra\u00adkis\u00ad(4-meth\u00adoxy\u00adphen\u00adyl)porphyrinate ligand and two water mol\u00adecules. The average equatorial Fe\u2014N(pyrrole) bond length [2.043\u2005(6)\u2005\u00c5] is consistent with a high-spin (S = 5/2) iron(III) metalloporphyrin derivative. The K+ cation, which also lies on an inversion centre, is chelated by the six O atoms of one 18-crown-6 mol\u00adecule and is additionally coordinated by two water mol\u00adecules in a distorted hexa\u00adgonal\u2013bipyramidal geometry. In the crystal, the cations, anions and one non-coordinating 18-crown-6 mol\u00adecule are linked by classical O\u2014H\u22efO hydrogen bonds and non-conventional C\u2014H\u22efO hydrogen bonds, leading to a one-dimensional supra\u00admolecular architecture along [10-1]. The crystal packing is further stabilized by weak C\u2014H\u22ef\u03c0 inter\u00adactions involving pyrrole and phenyl rings of the porphyrins, as well as weak C\u2014H\u22efF contacts involving the (SO3CF3)\u2212 counter-ion and the 18-crown-6 mol\u00adecules.In the title compound, [Fe DOI: 10.1107/S2056989015021039/im2471Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015021039/im2471fig1.tifORTEP III 2 2 + 2 2 + 3 3 \u2212 x y z . DOI: ORTEP view of the [FeIII(TMPP)(H2O)2]+ and [K(18-crown-6)(H2O)2]+ cations, the (SO3CF3)\u2212 anion and the 18-crown-6 mol\u00adecule. Displacement ellipsoids are drawn at 50% probability level. H atoms have been omitted for clarity. .An Click here for additional data file.10.1107/S2056989015021039/im2471fig2.tifb Cg . DOI: b axis. The O\u2014H\u22efO classic H bonds are drawn as dashed light blue lines, the C\u2014H\u22efO contacts as dashed dark red lines while the C\u2014H\u22efCg inter\u00admolecular inter\u00adactions are shown as dashed green lines.A drawing showing the one-dimensional supra\u00admolecular structure of the title compound viewed down the 976053CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Two novel G3P[4] rotavirus strains were detected from children with acute diarrhea in Sendai, Japan, identified as a G3\u2013P[4]\u2013I2\u2013R2\u2013C2\u2013M2\u2013A2\u2013N2\u2013T2\u2013E2\u2013H2 genotype constellation by whole-genome sequence analysis. The VP7 gene of the two strains displayed the highest nucleotide sequence identity (91\u00a0%) and showed a close genetic relationship to an equine rotavirus reported in India. The other gene segments were related to human group A rotaviruses. This report suggests a possible reassortment event between human and equine rotaviruses.The online version of this article (doi:10.1007/s11262-014-1135-z) contains supplementary material, which is available to authorized users. Rotaviruses are a leading cause of acute diarrhea in young children and cause approximately 453,000 deaths per year worldwide . In JapaReoviridae, genus Rotavirus. Its double-stranded RNA genome consists of 11 gene segments that encode six structural viral proteins and six non-structural proteins (NSP1\u20136). The genes of the two antigenically distinct outer capsid proteins, VP7 (G type) and VP4 (P type), are used for genotyping [x\u2013P[x]\u2013Ix\u2013Rx\u2013Cx\u2013Mx\u2013Ax\u2013Nx\u2013Tx\u2013Ex\u2013Hx (where \u201cx\u201d depicts the number of the genotype) for symbolizing the VP7\u2013VP4\u2013VP6\u2013VP1\u2013VP2\u2013VP3\u2013NSP1\u2013NSP2\u2013NSP3\u2013NSP4\u2013NSP5 genes followed by G3P[8], G9P[8], and G2P[4] . Rotavir\u00b0C for 5\u00a0min, and cooled on ice. Then 2\u00a0\u03bcl of 5\u00d7\u00a0first strand buffer (F.C: 1\u00d7), 0.5\u00a0\u03bcl of 0.1\u00a0M DTT (F.C: 5\u00a0mM), 0.25\u00a0\u03bcl of 40U/\u03bcl RNaseOUT (F.C: 1U/\u03bcl), and 0.25\u00a0\u03bcl of 200\u00a0U/\u03bcl superscript III RT (F.C: 5\u00a0U/\u03bcl) were added into the mixture. The reverse transcription step was carried out at 25\u00a0\u00b0C for 10\u00a0min, 50\u00a0\u00b0C for 50\u00a0min, followed by heating at 70\u00a0\u00b0C for 15\u00a0min to inactivate the enzyme, and cooling at 4\u00a0\u00b0C immediately. For screening of RVA, real-time PCR was performed , and 0.5\u00a0\u03bcl of 300\u00a0ng/\u03bcl random hexamer primers (F.C: 15\u00a0ng/\u03bcl) and 0.5\u00a0\u03bcl of 10\u00a0mM dNTP (F.C: 0.5\u00a0mM), heated at 98\u00a0erformed and for erformed . Nucleothttp://rotac.regatools.be/) . Interestingly, two samples were of the G3P[4] genotype. One of the samples (S13\u201330) was collected from a 4-year-old boy in the last week of February 2013 and the other (S13\u201345) was collected from a 1-year-old infant girl in the middle of March 2013. Both children did not receive rotavirus vaccines.In Table\u00a0Phylogenetic trees of all 11 segments were constructed based on complete nucleotide sequences of each gene Below is the link to the electronic supplementary material."} +{"text": "Acridine\u2013acridine stacking, thio\u00adphene\u2013thio\u00adphene stacking and acridine\u2013thio\u00adphene C\u2014H\u22ef\u03c0 inter\u00adactions also occur in the crystal.In the title co-crystal, C The inter\u00adnal angle at N1 [C6\u2014N1\u2014C18 = 119.30\u2005(15)\u00b0] and bond lengths [C18\u2014N1 = 1.346\u2005(2) and C6\u2014N1 = 1.354\u2005(2)\u2005\u00c5] agree with those reported for neutral acridine structures , Cg1\u22efCg1ii = 3.7526\u2005(9), Cg1\u22efCg2ii = 3.7293\u2005(12), Cg2\u22efCg3i = 3.6748\u2005(12) and Cg2\u22efCg3ii = 3.7298\u2005(12)\u2005\u00c5 where Cg1 is the centroid of the N1/C6/C11/C12/C13/C18 ring, Cg2 is the centroid of the C6\u2013C11 ring and Cg3 is the centroid of the C13\u2013C18 ring; symmetry codes: (i) \u2212x, 2\u00a0\u2212\u00a0y,1\u00a0\u2212\u00a0z; (ii) 1\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y,1\u00a0\u2212\u00a0z] and between the thio\u00adphene rings . The crystal structure also features C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional supra\u00admolecular architecture (Table\u00a01The 3TPC and acridine moieties are linked by an O\u2014H\u22efN hydrogen-bonding inter\u00adaction between (O1\u2014H1) of the carboxyl group and the acridine nitro\u00adgen atom (N1) Table\u00a01. This O\u2014e Table\u00a01.et al., 2001aet al., 2014et al., 2011et al., 2001bet al., 2011The crystal structures of a number of acridine co-crystals, acridinium salts and their metal complexes have been investigated in a variety of crystalline environments such as diphenic acid\u2013acridine (1:1) (Shaameri 1) were obtained.To 10\u2005ml of a hot methanol solution of 3TPC were added 10\u2005ml of a hot methano\u00adlic solution of acridine . The resulting solution was warmed over a water bath for half an hour and then kept at room temperature for crystallization. After a week yellow plate-like crystals of (Uiso(H) = kUeq, where k = 1.5 for hy\u00addroxy and 1.2 for all other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016005685/hg5473sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016005685/hg5473Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016005685/hg5473Isup3.cmlSupporting information file. DOI: 1472507CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure is formed by an alternating packing of organic and inorganic layers along [001] and contains isolated (Bi2Cl10)4\u2212 bi\u00adocta\u00adhedra.The title salt, (C 6H14N2)2[Bi2Cl10]\u00b72H2O, was ob\u00adtain\u00aded by slow evaporation at room temperature of a hydro\u00adchloric aqueous solution (pH = 1) containing bis\u00admuth(III) nitrate and 1,4-di\u00adaza\u00adbicyclo\u00ad[2.2.2]octane (DABCO) in a 1:2 molar ratio. The structure displays a two-dimensional arrangement parallel to (100) of isolated [Bi2Cl10]4\u2212 bi\u00adocta\u00adhedra (site symmetry -1) separated by layers of organic 1,4-diazo\u00adniabi\u00adcyclo\u00ad[2.2.2]octane dications [(DABCOH2)2+] and water mol\u00adecules. O\u2014H\u22efCl, N\u2014H\u22efO and N\u2014H\u22efCl hydrogen bonds lead to additional cohesion of the structure.The title compound bis\u00ad di-\u03bc-chlorido-bis\u00ad[tetra\u00adchlorido\u00adbis\u00admuthate(III)] dihydrate, (C These components are linked by strong hydrogen bonds. The inorganic moiety is an edge-sharing di\u00adocta\u00adhedron located site with symmetry 2)2+ dications to 176.64\u2005(15)\u00b0 for the trans arrangement. Using Shannon\u2019s method 2[Bi2Cl10]\u00b72H2O structure is very close to that of (C6H14N2)2[Sb2Cl10]\u00b72H2O in the Pnnm anti\u00admony unit cell to be comparable to the bis\u00admuth one \u00d7 10\u22123] . It is worth noting that the water mol\u00adecule plays a more efficient role in the bis\u00admuth based compound. In (C6H14N2)2[Sb2Cl10]\u00b72H2O, the H2O mol\u00adecules are only linked to (DABCOH2)2+ and in the (C6H14N2)2[Bi2Cl10]\u00b72H2O structure they are directly hydrogen bonded to both the organic and inorganic parts \u2005\u00c5] because the metallic coordination polyhedra are aligned along this axis. On the other hand, a roughly equivalent decrease of the b parameter is observed causing the unit-cell volume of the two compounds approximately to be the same. A general comparison of the two structures reveals that they have a similar 3D pattern, built up by isolated bi\u00adocta\u00adhedra, (DABCOH2)2+ cations and water mol\u00adecules leaving empty the same voids. On the other hand, the water mol\u00adecule immediate environment is more regular in the Sb structure and the (DABCOH2)2+ cation is more distorted in the Bi structure explaining the lowering of the symmetry in the title compound.The x, \u2212y\u00a0+\u00a00.5, z\u00a0+\u00a00.5] and O\u2014HW2\u22efCl5 inter\u00adactions. The DABCO cations are hydrogen bonded to water mol\u00adecules, leading to chains composed of organic moieties, inorganic clusters and H2O mol\u00adecules running along the b direction 2[Bi2Cl10]\u00b72H2O crystals were obtained at ambient conditions by dissolving Bi(NO3)3\u00b75H2O and DABCO (C6H12N2) in water in a 1:2 molar ratio. The pH of the solution was adjusted to 1 with HCl. The mixture was stirred and kept for several days. Colourless crystals were obtained after a few weeks. I. DOI: 10.1107/S2056989015019933/vn2102Isup2.hklStructure factors: contains datablock(s) I. DOI: 971956CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "B-Spline curves. Since modal intervals are used in many fields, we introduce a new sequence space h(gI) called the Hahn sequence space of modal intervals. We have given some new definitions and theorems. Some inclusion relation and some topological properties of this space are investigated. Also dual spaces of this space are computed.The history of modal intervals goes back to the very first publications on the topic of interval calculus. The modal interval analysis is used in Computer graphics and Computer Aided Design (CAD), namely, the computation of narrow bounds on Bezier and Goes and S. Goes \u03b2\u03b2.(ii)E(gI)]\u03b2\u03b2\u03b2 = [E(gI)]\u03b2.[(iii)E1(gI)\u2283E(gI) then [E1(gI)]\u03b2 \u2282 [E(gI)]\u03b2.If Let The same results hold for dual also.Let (i) Suppose E(gI). Hence E(gI)\u2282[E(gI)]\u03b2\u03b2. The proof for (ii) follows similarly.This means that E1(gI)\u2283E(gI), E1(gI)]\u03b2 \u2282 [E(gI)]\u03b2.(iii) Suppose B-transform of a sequence Define the sequence l\u221e(gI) is the set of all sequences such that the B-transforms of them are in l\u221e(gI). That is, That is, \u03c3(l\u221e(gI)) is a complete metric space with the metric \u03c3(l\u221e(gI)).\u03c3(l\u221e(gI)) where \u03b5 > 0, there exists a positive integer n0(\u03b5) such that k \u2208 N from that i \u2265 n0(\u03b5), taking limit j \u2192 \u221e first and letting m \u2192 \u221e in (\u03c3(l\u221e(gI)), we have Consider the sequence ave from for eachm \u2192 \u221e in , we obtaNow, \u03c3(l\u221e(gI)) is complete.Since \u03b2-dual and \u03b3-dual of h(gI) are \u03c3(l\u221e(gI)).The \u03b5 > 0, there exists n0 such that Let k and n.Since M > 0 such that k and n.Hence there exists an M-test.Now, k \u2208 N.Thus h(gI)]\u03b2 = \u03c3(l\u221e(gI)). This completes the proof.Hence gI. From and 31)(31)v~k\u2208\u03c3Considerh(gI) \u2282 l1(gI)\u2229\u222bc0(gI).(i) h(gI) = l1(gI)\u2229\u222bbv(gI) = l1(gI)\u2229\u222bbv0(gI).(ii) (i) Let Consider And also since h(gI) \u2282 l1(gI)\u2229\u222bc0(gI).Therefore, k = 1,2,\u2026, (ii) For h(gI) \u2282 l1(gI). Hence also h(gI) \u2282 \u222bbv(gI).The last series is convergent since Hence versely, implies h(gI) = l1(gI)\u2229\u222bbv(gI).Therefore, nce from and 36)(36)l1gI\u2229Similarly, we can prove other equalities."} +{"text": "II atom coordinated by two Cl atoms and two dimethyl N-cyano\u00addithio\u00adimino\u00adcarbonate ligands bonded through the terminal N atom in a distorted tetra\u00adhedral manner. The complex mol\u00adecules inter\u00adact through C\u2014H\u22efCl and Cl\u22efS inter\u00adactions to give a layered structure in the crystal.The title complex consists of a Zn II atom in the title complex, [ZnCl2(C4H6N2S2)2], is coordinated in a distorted tetra\u00adhedral manner by two Cl atoms and two terminal N atoms of two dimethyl N-cyano\u00addithio\u00adimino\u00adcarbonate ligands. In the crystal, the complex mol\u00adecules are connected through C\u2014H\u22efCl hydrogen bonds and Cl\u22efS contacts, leading to a two-dimensional structure extending parallel to the ab plane.The Zn N-cyano\u00addithio\u00adimino\u00adcarbonate (DMCDIC), which are expected to act as hard and soft donors, respectively, according to Pearson\u2019s concept, give an inter\u00adesting coordination potential to this mol\u00adecule. However, only one structure of a metal complex with DMCDIC acting as a ligand has been reported 2] ; \u03c44 = [360 - (\u03b1 + \u03b2)]/141, where \u03b1 and \u03b2 are the two largest tetra\u00adhedral angles.The structure of the title compound Fig.\u00a01 is isotyB\u22efCl1ii and C7\u2014H7B\u22efCl1ii; Table\u00a01B\u22efCl2i hydrogen bond (Table\u00a01i short contact [3.3812\u2005(7)\u2005\u00c5], giving infinite chains along , leading to a layer parallel to the ab plane .All chemicals are purchased from Aldrich Company, Germany and used as received. Dimethyl cyano\u00adcarbonimidodi\u00adthio\u00adate was mixed in aceto\u00adnitrile with ZnCl2CNCN)2Cl2] = 1.5Ueq(C), and were allowed to rotate to minimize their contribution to the electron density.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016002607/is5443sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016002607/is5443Isup2.hklStructure factors: contains datablock(s) I. DOI: 1453191CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound, an aryl\u00adsulfonyl glycinyl aryl hydrazone Schiff base, crystallizes with two independent mol\u00adecules in the asymmetric unit. In the crystal, a series of N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 and slipped parallel \u03c0\u2013\u03c0 inter\u00adactions link the mol\u00adecules, forming a three-dimensional structure. 17H19N3O3S, containing an amino acid moiety and electron-donating substituents attached to both the phenyl rings, crystallized with two independent mol\u00adecules (A and B) in the asymmetric unit. The mol\u00adecules are bent at the S atom, with C\u2014SO2\u2014NH\u2014CH2 torsion angles of \u221267.3\u2005(2) and 67.7\u2005(3)\u00b0 in mol\u00adecules A and B, respectively. Further, the dihedral angles between the sulfonyl\u00adglycine segments and the p-toluene\u00adsulfonyl rings are 76.1\u2005(1) and 85.8\u2005(1)\u00b0 in mol\u00adecules A and B, respectively. The central hydrazone segments and the toluene rings attached to them are almost co-planar with their mean planes being inclined to one another by 5.2\u2005(2) (mol\u00adecule A) and 2.9\u2005(2)\u00b0 (mol\u00adecule B). The dihedral angles between the benzene rings are 86.83\u2005(12) (mol\u00adecule A) and 74.00\u2005(14)\u00b0 (mol\u00adecule B). In the crystal, the A mol\u00adecules are linked by a pair of N\u2014H\u22efO hydrogen bonds, forming inversion dimers with an R22(8) ring motif. The dimers are linked via three N\u2014H\u22efO hydrogen bonds involving the B mol\u00adecules, forming chains along [100] and enclosing R22(12) and R44(16) ring motifs. The chains are linked via C\u2014H\u22efO hydrogen bonds and a C\u2014H\u22ef\u03c0 inter\u00adaction, forming sheets parallel to (010). There is a further C\u2014H\u22ef\u03c0 inter\u00adaction and a slipped parallel \u03c0\u2013\u03c0 inter\u00adaction [inter-centroid distance = 3.8773\u2005(16)\u2005\u00c5] between the sheets, leading to the formation of a three-dimensional framework.The title acyl\u00adhydrazone derivative, C N-acyl\u00adhydrazones containing a glycine residue have been investigated extensively for their biological and medical activities due to the presence of the C=O group, an additional donor site. N-(ar\u00adyl)-amides of the title compound are shown in Fig.\u00a01A has an extended conformation while mol\u00adecule B is U-shaped. In mol\u00adecule A, the conformations of the hydrazide N\u2014H and C\u2014H bonds are syn to each other, while the N\u2014H and C=O bonds are anti to each other. On the sulfonamide side, the conformations of the sulfonamide N\u2014H and C=O bonds are syn to each other. In mol\u00adecule B, the conformations of the hydrazide N\u2014H and C\u2014H bonds, the hydrazide N\u2014H and C=O, and the C=O and sulfonamide N\u2014H bonds are all syn to each other.The mol\u00adecular structures of the two independent mol\u00adecules \u00b0. The mean plane through atoms C9/N3/N2/C8/O3/C7 [maximum deviation of 0.043\u2005(2)\u2005\u00c5 for N2], the central section of the mol\u00adecule, is inclined to the two benzene rings, C1\u2013C6 and C10\u2013C15, by 86.38\u2005(12) and 7.22\u2005(12)\u00b0, respectively. In mol\u00adecule B, the benzene rings (C18\u2013C23 and C27\u2013C32) are inclined to one another by 74.00\u2005(14)\u00b0, and by 76.85\u2005(13) and 2.91\u2005(12)\u00b0, respectively, to the mean plane through atoms C26/N6/N5/C25/O6/C24 [maximum deviation of 0.061\u2005(2)\u2005\u00c5 for C26]. The different conformations of mol\u00adecules A and B are further demonstrated by the differences in the equivalent torsion angles; N1\u2014C7\u2014C8\u2014N2 = 29.3\u2005(3) \u00b0 in A, compared to N4\u2014C24\u2014C25\u2014N5 = 177.2\u2005(2)\u00b0 in B, and C1\u2014S1\u2014N1\u2014C7 = \u221267.3\u2005(2)\u00b0 in A, compared to C18\u2014S2\u2014N4\u2014C24 = 67.7\u2005(3)\u00b0 in B.In mol\u00adecule A and C25\u2014O6 in B, are 1.214\u2005(3) and 1.229\u2005(3)\u2005\u00c5, respectively, indicating that the mol\u00adecules exist in the keto form in the solid state. The C9=N3 and C26=N6 bond lengths, both 1.272\u2005(3)\u2005\u00c5 in mol\u00adecules A and B, respectively, confirm their significant double-bond character. The N2\u2014N3 and N5\u2014N6 bond distances are 1.383\u2005(3) and 1.379\u2005(3)\u2005\u00c5, respectively, and the C8\u2014N2 and C25\u2014N5 bond distances are 1.339\u2005(3) and 1.334\u2005(3)\u2005\u00c5, respectively, which indicates significant delocalization of \u03c0-electron density over the hydrazone portions of the mol\u00adecules.The carbonyl bonds lengths, C8\u2014O3 in A mol\u00adecules are linked by a pair of N\u2014H\u22efO hydrogen bonds, forming inversion dimers with an via three N\u2014H\u22efO hydrogen bonds involving the B mol\u00adecules, forming chains along [100] that enclose via C\u2014H\u22efO hydrogen bonds and a C\u2014H\u22ef\u03c0 inter\u00adaction, forming sheets parallel to (010). The is a C\u2014H\u22ef\u03c0 inter\u00adaction and a slipped parallel \u03c0\u2013\u03c0 inter\u00adaction , between the sheets, leading to the formation of a three-dimensional framework \u2013NH\u2013N=CH\u2013 yielded only one hit, namely N-(2-hy\u00addroxy-1-naphthyl\u00admethyl\u00adene)-N\u2032-(N-phenyl\u00adglyc\u00adyl)hydrazine -N-{2-[2-(3-chloro\u00adbenzyl\u00adidene) hydrazin\u00adyl]-2-oxoeth\u00adyl}-4-methyl\u00adbenzene\u00adsulfonamide monohydrate was added to glycine (0.02\u2005mol) dissolved in an aqueous solution of potassium carbonate . The reaction mixture was stirred at 373\u2005K for 6\u2005h, left overnight at room temperature, then filtered and treated with dilute hydro\u00adchloric acid. The solid N-(p-toluene\u00adsulfon\u00adyl)glycine (L1) obtained was crystallized from aqueous ethanol.L1 (0.02\u2005mol) dissolved in ethanol (30\u2005ml) and the mixture was refluxed. The reaction was monitored by TLC at regular inter\u00advals. After completion of the reaction, the reaction mixture was concentrated to remove the excess ethanol. The product, N-(p-toluene\u00adsulfon\u00adyl)glycine ethyl ester (L2) was poured into water, neutralized with sodium bicarbonate and recrystallized from acetone.Sulfuric acid (0.5\u2005ml) was added to L2 (0.01\u2005mol) was then added in small portions to a stirred solution of 99% hydrazine hydrate (10\u2005ml) in 30\u2005ml ethanol and the mixture was refluxed for 6\u2005h. After cooling to room temperature, the resulting precipitate was filtered, washed with cold water and dried to give N-(p-toluene\u00adsulfon\u00adyl)glycinyl hydrazide (L3).The pure L3 (0.01\u2005mol) and p-methyl\u00adbenzaldehyde (0.01\u2005mol) in anhydrous methanol (30\u2005ml) and two drops of glacial acetic acid was refluxed for 8\u2005h. After cooling, the precipitate was collected by vacuum filtration, washed with cold methanol and dried. It was recrystallized to constant melting point from methanol (455\u2013457\u2005K). Prism-like colourless single crystals were grown from a DMF solution by slow evaporation of the solvent. The purity of the compound was checked by TLC and characterized by its IR spectrum. The characteristic absorptions observed are 3286.7, 1678.1, 1606.7, 1323.2 and 1157.3\u2005cm\u22121 for the stretching bands of N\u2014H, C\u2014O, C\u2014N, S\u2014O asymmetric and S\u2014O symmetric, respectively.A mixture of Uiso(H) = 1.2Ueq(N). The C-bound H atoms were positioned with idealized geometry and refined using a riding model: C\u2014H = 0.93\u20130.97\u2005\u00c5 with Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015009330/su5131sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015009330/su5131Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015009330/su5131Isup3.cmlSupporting information file. DOI: 1401257CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Deionised water (negative) and Solcoseryl (positive) were applied on the control groups. Histochemical and biochemical assays were used to evaluate cellular and bio-molecular changes in the wound.Excision wounds were created on the dorsum of Sprague Dawley rats and treated topically twice/day with 20\u00a0\u03bcL of PE and WE (5\u00a0mg extract/mL), 0.5 \u03bcgSe2+) only increased significantly (p\u2009<\u20090.05) wound total protein content on day 10 post wounding. PES increased significantly (p\u2009<\u20090.05) the number of fibroblasts/high power field (HPF) (75.60\u2009\u00b1\u20099.66) but decreased significantly (p\u2009<\u20090.05) the number of polymorphonuclear leukocytes/HPF (59.20\u2009\u00b1\u200912.64) in the wound compared to positive control on day 4. Similar results were recorded for WES. PES demonstrated increased neovascularization, TGF-\u03b21 and VEGFA expressions at day 4 and increased collagen at day 10.PES (PE\u2009+\u20090.5\u00a0\u03bcg Se2+ addition.Papaya extract improved wound repair by increasing fibroblasts recruitment and reducing polymorphonuclear leukocytes infiltration through early transient expressions of TGF-\u03b21 and VEGFA at the wound area. The processes were amplified with SeThe online version of this article (doi:10.1186/s12906-015-0900-4) contains supplementary material, which is available to authorized users. Carica papaya L (papaya) has been attributed to its high content of cystein endopeptidase, mineral nutrients and vitamins .2O); gently shaken at 37\u00a0\u00b0C for 8\u00a0h, centrifuged at 200\u2009\u00d7\u2009g for 30\u00a0min at 4\u00a0\u00b0C and finally freeze-dried . Glucosamine hydrochloride (0.05\u00a0mg) was used as a standard. Values were corrected to the initial wound tissue dry weight.Wound tissue samples were completely dried in oven at 60\u00a0\u00b0C and ground to powder. The HAM content was determined at 530\u00a0nm following the established protocol . A correp\u2009<\u20090.05.Data were analyzed using the statistical software SPSS . Data were expressed as mean\u2009\u00b1\u2009standard error of mean (SEM). One way-Anova was used to analyse all data and Tukey\u2019s post-hoc test was used to find the mean differences within the groups at different time interval. Data were considered statistically significant at p\u2009<\u20090.05) increase in the total protein content of the wound tissue (95.14\u2009\u00b1\u20091.15\u00a0mg/g tissue) as compared to the treatment with PC (80.42\u2009\u00b1\u20090.86\u00a0mg/g tissue). In contrast, Zn added extracts did not show any significant changes in the wound total protein content on day 4 and day 10 post wounding as compared to NC increase in HOP content (20.07\u2009\u00b1\u20093.19\u00a0\u03bcg/mg tissue and 19.80\u2009\u00b1\u20092.51\u00a0\u03bcg/mg tissue respectively) on day 4 compared to NC (8.47\u2009\u00b1\u20091.50\u00a0\u03bcg/mg tissue) and significantly (p\u2009<\u20090.05) increase HOP content (55.15\u2009\u00b1\u20091.06\u00a0\u03bcg/mg tissue and 52.61\u2009\u00b1\u20091.02\u00a0\u03bcg/mg tissue respectively) on day 10 compared to NC (25.35\u2009\u00b1\u20091.04\u00a0\u03bcg/mg tissue), PC (40.62\u2009\u00b1\u20090.60\u00a0\u03bcg/mg tissue), PE (37.74\u2009\u00b1\u20091.24\u00a0\u03bcg/mg tissue) and WE (27.25\u2009\u00b1\u20091.03\u00a0\u03bcg/mg tissue). Treatment with PEZ and WEZ also resulted in significant (p\u2009<\u20090.05) increase in HOP content (30.52\u2009\u00b1\u20091.23\u00a0\u03bcg/mg tissue and 31.32\u2009\u00b1\u20091.34\u00a0\u03bcg/mg tissue respectively) on day 10 compared to NC (25.35\u2009\u00b1\u20091.04\u00a0\u03bcg/mg tissue) (Table\u00a0p\u2009<\u20090.05) after treatment with PE (53.41\u2009\u00b1\u20096.21\u00a0mg/g tissue), PES (60.84\u2009\u00b1\u20096.08\u00a0mg/g tissue) and WES (53.81\u2009\u00b1\u20097.36\u00a0mg/g tissue) compared to NC (20.71\u2009\u00b1\u20094.36\u00a0mg/g tissue) and WE (25.45\u2009\u00b1\u20095.32\u00a0mg/g tissue) on day 4 and only after treatment with PES (18.12\u2009\u00b1\u20093.32\u00a0mg/g tissue) compared to NC (5.82\u2009\u00b1\u20091.28\u00a0mg/g tissue) on day 10. Treatment with only PES resulted in significantly increased (p\u2009<\u20090.05) HAM content (35.23\u2009\u00b1\u20094.95\u00a0mg/g tissue and 18.21\u2009\u00b1\u20093.03\u00a0mg/g tissue) both on day 4 and day 10 respectively compared to NC (12.54\u2009\u00b1\u20094.53\u00a0mg/g tissue day 4 and 4.59\u2009\u00b1\u20091.57\u00a0mg/g tissue day 10) (Table\u00a0Treatment with PES and WES resulted in significant (p\u2009<\u20090.05) lower both on day 4 of the mean number of PMNL/HPF compared to NC (164.40\u2009\u00b1\u200925.93), PC (101.00\u2009\u00b1\u200927.99) and WE (126.00\u2009\u00b1\u200923.53) on day 4 higher on day 4 in all treatment groups compared to NC (5.60\u2009\u00b1\u20093.21). However, significantly increased (p\u2009<\u20090.05) number of fibroblasts/HPF was observed on day 4 in PES (75.60\u2009\u00b1\u20099.66) treated only group compared to that of PC, PE and WE groups when treated with PES (69.60\u2009\u00b1\u200913.07) as compared to NC (40.80\u2009\u00b1\u20099.09) Fig.\u00a0. On day 09) Fig.\u00a0.On day 4, wound tissue sections from all groups exhibited disrupted wound-dermis interface, with the absence of overlying epithelia which are affected due to injury [2+ supplementation was shown to improve healing of the wound caused by surgical incision, trauma, and burns [2+ added unripe C. papaya improves healing of excision wound through enhanced anti-inflammatory and antioxidant effect [Sentiation , 21. Whiferation . Both Seo injury . In reland burns . Earliert effect .The present study used female rats for the experimentation. Female rats are known to exhibit shorter period for complete wound healing and this has been linked to the circulating oestrogen level \u201326. Thou2+ added papaya mediates improved wound healing by reducing inflammation associated oxidative damage apparently via cyclooxygenase specific inhibition, arginine metabolism and up-regulation of antioxidant enzymes. These could be linked to the added Se2+ [Depending on the type and aetiology of wound, wound healing involves inflammation (day 1\u20134 or more), proliferative/repair day 5\u201310 or 11), and remodelling phases (day 11\u201321 or more) , 10. Pro or 11, aded Se2+ .2+ added PE/WE treated groups albeit the reduction was not as significant as Se2+ added PE/WE treated groups and biochemical events , a model is proposed Fig.\u00a0 to descrPE through mechanisms associated with cyclooxygenase and prostaglanding E2 inhibition reduced Consistently, PE induced a transient up-regulation of TGF-\u03b21 and VEGFA on day 4 which in turn enhanced cellular activity as evidenced in the increased fibroblasts recruitment and early appearance of new blood vessels in PE treatment. These effect was more pronounced in Se added PE.In addition, the establishment of new blood vessels enhanced nutrient delivery to the fibroblasts and enhanced fibroblastic synthesis of provisional matrix, primary molecules in the extracellular matrix and collagen. In line with this, Se added PE exclusively enhanced the synthesis of primary molecules of ground substances i.e. HUA and HAM. PE alone showed little effect in these regard. However, hydroxyproline an index for collagen turnover was found to be markedly increased, later on day 10, with PE treatment. Concurrently, well aligned collagen fibres and reconstructed dermis and epidermis were observed later on day 10 after treatment with PE and PES especially. Collectively, the increased recruitment of fibroblasts and their bio-molcular products resulted in the significant increase in wound total protein content later on day 10 in PE treated group.2+\u00a0added papaya extracts might (i) reduce PMNL infiltration to speed up the inflammatory phase as well as (ii) induced transient up-regulation of TGF-\u03b21 and VEGFA expression to enhance angiogenesis and fibroblast recruitment thereby enhanced cutaneous wound repair.Se"} +{"text": "III ion in the title complex is coordinated by two 1,10-phenanthroline (phen) ligands, one water mol\u00adecule and a chloride in a cis geometry, displaying a distorted octa\u00adhedral environment. The [ZnCl4]2\u2212 anion has a slightly distorted tetra\u00adhedral coordination geometry.The Cr 12H8N2)2(H2O)][ZnCl4]\u00b7H2O, has been determined from synchrotron data. The CrIII ion is bonded to four N atoms from two 1,10-phenanthroline (phen) ligands, one water mol\u00adecule and a Cl atom in a cis arrangement, displaying an overall distorted octa\u00adhedral coordination environment. The Cr\u2014N(phen) bond lengths are in the range of 2.0495\u2005(18) to 2.0831\u2005(18)\u2005\u00c5, while the Cr\u2014Cl and Cr\u2014(OH2) bond lengths are 2.2734\u2005(7) and 1.9986\u2005(17)\u2005\u00c5, respectively. The tetra\u00adhedral [ZnCl4]2\u2212 anion is slightly distorted owing to its involvement in O\u2014H\u22efCl hydrogen bonding with coordinating and non-coordinating water mol\u00adecules. The two types of water mol\u00adecules also inter\u00adact through O\u2014H\u22efO hydrogen bonds. The observed hydrogen-bonding pattern leads to the formation of a three-dimensional network structure.The structure of the title compound, [CrCl(C These complexes are promising materials for the development of new mol\u00adecule-based magnets, solar energy storage media or tunable solid state lasers ][ZnCl4]\u00b7H2O , (I)We report here on the synthesis and crystal structure of the title compound, (ClO4)3\u00b7H2O 2]ClO4\u00b7H2O 2]Cl bond lengths in (I)2) bond length is comparable to those of 1.947\u2005(4), 1.9579\u2005(10) and 1.996\u2005(4)\u2005\u00c5 found in cis-[Cr(dpp)(phen)2(H2O)](NO3)2\u00b7H2O\u00b7CH3CN [Hdpp = (C6H5O)2\u00b7PO2H] 2(H2O)](ClO4)2\u00b72H2O (H2O)](ClO4)2\u00b7H2O 2]Cl 2Cl2]Cl A\u2014Cr1A\u2014N2A and N3A\u2014Cr1A\u2014N4A are 79.76\u2005(5) and 80.23\u2005(7)\u00b0.The Cr\u2014N(phen) bond lengths are in the range of 2.0495\u2005(18) to 2.0831\u2005(18)\u2005\u00c5 and are in good agreement with those observed in 2\u2212 anion and the second water mol\u00adecule remain outside the coordination sphere. The ZnII atom in the complex anion exhibits a slightly distorted tetra\u00adhedral coordination sphere caused by the influence of hydrogen bonding on the Zn\u2014Cl bond lengths and the Cl\u2014Zn\u2014Cl angles. The Zn\u2014Cl bond lengths range from 2.2443\u2005(7) to 2.2854\u2005(7)\u2005\u00c5 and the Cl\u2014Zn\u2014Cl angles from 107.54\u2005(4) to 111.57\u2005(3)\u00b0.The [ZnClB of the [ZnCl4]2\u2212 anion and the Cl1A ligand atom are not involved in hydrogen bonding. An extensive array of O\u2014H\u2014O and O\u2014H\u22efCl contacts (phen)2(H2O)](NO3)2\u00b7H2O\u00b7CH3CN 3\u00b7H2O 2]ClO4 2]Cl ]2+ with any anions have been deposited.A search of the Cambridge Structural Database 2]ClO4\u00b7H2O was prepared according to a literature procedure 2]ClO4\u00b7H2O (0.2\u2005g) was dissolved in 10\u2005mL of 0.01\u2005M HCl at 313\u2005K, and 5\u2005mL of 1\u2005M HCl containing 1.2\u2005g of solid ZnCl2 were added to this solution. The mixture was refluxed at 328\u2005K for 30\u2005min and then cooled to room temperature. The resulting solution was filtered and allowed to stand at room temperature for 3\u20135 days, giving purple crystals of (I)All chemicals were reagent-grade materials and used without further purification. The starting material, Uiso(H) set to 1.2Ueq(C). The H atoms of water mol\u00adecules were located from difference Fourier maps and refined with restraints and an O\u2014H distance of 0.84\u2005(1)\u2005\u00c5, with Uiso(H) values of 1.2 Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015003266/wm5123sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015003266/wm5123Isup2.hklStructure factors: contains datablock(s) I. DOI: 1049598CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of the title compound consists of discrete octa\u00adhedral cobalt(II) complexes that are linked by a variety of hydrogen-bonding inter\u00adactions into a three-dimensional network. 2(C6H6N2O)4]\u00b72C6H6N2O\u00b7C2H5OH, comprises one CoII cation, two thio\u00adcyanate anions, four coordinating and two solvent isonicotinamide molecules and one ethanol solvent mol\u00adecule. The CoII cations are octa\u00adhedrally coordinated by four N-coordinating isonicotinamide ligands and two terminally N-bonded thio\u00adcyanate anions. These discrete complexes are linked by inter\u00admolecular N\u2014H\u22efO and N\u2014H\u22efS hydrogen-bonding inter\u00adactions into a three-dimensional network. The two isonicotinamide and the ethanol solvent mol\u00adecules are embedded in channels of this network and are linked through further N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds to the network. The ethanol solvent mol\u00adecule is disordered over two sets of sites (occupancy ratio 0.6:0.4).The asymmetric unit of the title compound, [Co(NCS) However, instead of the expected chain compound, a discrete complex with additional solvate mol\u00adecules of composition [Co(NCS)2(C6H6N2O)4]\u00b72C6H6N2O\u00b7C2H5OH was obtained in the current study and characterized by single-crystal X-ray diffraction.There is an increasing inter\u00adest in compounds showing cooperative magnetic properties, such as ferromagnetism, anti\u00adferromagnetism and metamagnetism or a slow relaxation of the magnetization, indicative of single-mol\u00adecule or single-chain magnetism and one positionally disordered ethanol solvent mol\u00adecule. The CoII cation is coordinated by two terminal N-bonded thio\u00adcyanate anions and four N-coordinating isonicotinamide ligands, forming a slightly distorted octa\u00adhedron \u2013178.32\u2005(11)\u00b0 for trans angles] are indicative for a slight distortion and are comparable with those in similar coordination compounds with CoII, thio\u00adcyanate anions and N-bound co-ligands.The asymmetric unit of the title compound consists of one Coon Fig.\u00a01. Bond lea axis by inter\u00admolecular N\u2014H\u22efO hydrogen-bonding inter\u00adactions with 73.3\u2005mg isonicotinamide (0.6\u2005mmol) in ethanol (1.5\u2005ml) after being allowed to stand for a few days at room temperature.Cobalt(II) thio\u00adcyanate and isonicotinamide were obtained from Alfa Aesar and were used without further purification. Single crystals suitable for structure analysis were obtained by the reaction of 26.3\u2005mg Co(NCS)Uiso(H) = 1.2Ueq (1.5 for methyl and O\u2014H hydrogen atoms) using a riding model with C\u2014H = 0.95\u2005\u00c5 for aromatic, C\u2014H = 0.98\u2005\u00c5 for methyl, N\u2014H = 0.88\u2005\u00c5 and O\u2014 H = 0.84\u2005\u00c5, respectively. The ethanol mol\u00adecule was found to be disordered over two sets of sites and was refined with fixed occupation factors of 0.6 and 0.4, respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016010951/wm5305sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016010951/wm5305Isup2.hklStructure factors: contains datablock(s) I. DOI: 1491089CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Within the complex, intra\u00admolecular C\u2014H\u22efS, C\u2014H\u22ef\u03c0, N\u2014H\u22efBr and \u03c0\u2013\u03c0 stacking inter\u00adactions are observed. In the crystal, the components are linked by N\u2014H\u22efBr and C\u2014H\u22efN hydrogen bonds and weak \u03c0\u2013\u03c0 stacking inter\u00adactions, generating chains propagating in the [100] direction.In the solvated dinuclear complex \u00b72C2H3N or [Cu2Br2(ATU)(dppm)2]\u00b72CH3CN. Both Cu+ ions adopt distorted tetra\u00adhedral geometries, being coordinated by one terminal Br atom, one \u03bc2-S atom of the bridging ATU ligand and two P atoms of the bridging dppm ligands. Within the complex, intra\u00admolecular C\u2014H\u22efS, C\u2014H\u22ef\u03c0, N\u2014H\u22efBr and \u03c0\u2013\u03c0 stacking inter\u00adactions are observed. In the crystal, the components are linked by N\u2014H\u22efBr and C\u2014H\u22efN hydrogen bonds and weak \u03c0\u2013\u03c0 stacking inter\u00adactions, generating chains propagating in the [100] direction.The reaction of cuprous bromide with a mixture of 1,1-bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)methane (dppm: C Both CuI atoms share a tetra\u00adhedral corner via an S-atom bridge.The title complex [Cu2I2(C3H8N2S)(C25H22P2)2]\u00b71.5CH3CN 3]\u00b7CH3CN 2P2 plane of the Cu1\u2013S1\u2013Cu2\u2013P3\u2013C38\u2013P4 chair conformation by an angle [72.59\u2005(3)\u00b0] that is significantly larger than that for the Cu1\u2013S1\u2013Cu2\u2013P1\u2013C13\u2013P2 plane [45.68\u2005(2)\u00b0]. Thus, the C\u2014H\u22efS effect might be the influence that leads to the longer distance between the centroids of the phenyl rings, Cg8\u22efCg9 [4.0356\u2005(17)\u2005\u00c5] compared with the Cg4\u22efCg5 distance [3.6097\u2005(19)\u2005\u00c5]. The intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction (C31\u2014H31\u22efCg6) also imposes a contact, of 3.476\u2005(3)\u2005\u00c5, between the Csp2 atom of the phenyl ring and another phenyl ring centroid (C20\u2013C25) of the other dppm ligand. An inter\u00adaction (C12\u2014H12\u22efN2) between Cg5 and the NH2 group is also observed. In addition, two intra\u00admolecular N\u2014H\u22efBr inter\u00adactions are found between the NH2 group of ATU and Br atoms. A perspective view of all the intra\u00admolecular inter\u00adactions in (I)The intra\u00admolecular inter\u00adaction C44\u2014H44\u22efS1 Table\u00a02 has an ex, 2\u00a0\u2212\u00a0y, \u2212z), which generates Cg8\u22efCg8iii rings generating a chain of alternating N\u2014H\u22efBr links and \u03c0\u2013\u03c0 stacking running along [100], as shown in Fig.\u00a03ii and C50\u2014H50\u22efN3i; symmetry codes: (i) \u2212x\u00a0+\u00a01, y\u00a0+\u00a0z\u00a0+\u00a0x, y\u00a0\u2212\u00a0z\u00a0+\u00a0In the crystal, neighbouring dinuclear mol\u00adecules form a hydrogen-bonded dimer held together by two N\u2014H\u22efBr bonds as a cyclic pattern with its symmetry\u2013equivalent partner, generated by a crystallographic inversion center (symmetry code: \u22122I2(C3H8N2S)(dppm)2]\u00b71.5CH3CN, studied by Nimthong et al. was added and further refluxed for four\u2005h and the precipitate slowly disappeared. Colorless crystals were obtained after the clear solution was left to evaporate at room temperature for a few days. The yield was 33% based on CuBr. Calculated for C58H58Br2Cu2N4P4S: C 55.55, H 4.66, N 4.47 and S 2.56%. Found: C 55.28, H 4.28, N 3.68 and S 2.54%. The main IR bands : 3282 [\u03bd(NH2)], 3168 [\u03bd(NH2)], 3058 [\u03bd(=C\u2014H)ph], 1580 [\u03bd(C=S)] and 1110 [\u03bd(P\u2014Cph)].Copper(I) bromide was added to a solution of 1,1-bis\u00ad(di\u00adphenyl\u00adphosphino)methane in 30\u2005ml aceto\u00adnitrile at 338\u2005K. The mixture was refluxed for two\u2005h and a white precipitate was formed. After that, Uiso(H) = 1.2Ueq(C) or 1.5Ueq(Cmethyl). The N\u2014H atoms were located in a difference map and the coordinates were refined with an N\u2014H distance restraint of 0.86\u2005\u00c5 and with Uiso(H) = 1.2eqU(N). Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015015637/hb7479sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015015637/hb7479Isup2.hklStructure factors: contains datablock(s) I. DOI: 1419827CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports6: Article number: 2482310.1038/srep24823; published online: 04212016; updated: 05312016This Article contains a typographical error in the Results section under subheading \u20185G2\u2009mAb recognized sugar chain epitopes\u2019,a GlcNAc), Lec , core1 , core2 GalNAc), and Sia\u03b12-6core1 GalNAc) structures .\u201d\u201cThe 5G2\u2009mAb recognized the Leshould read:a GlcNAc), Lec , core1 , core2 GalNAc), and Sia\u03b12-6core1 GalNAc) structures .\u201d\u201cThe 5G2\u2009mAb recognized the Le"} +{"text": "H)-one compound that has been designed and synthesized to explore a new electron-donor (D) and -acceptor (A) conjugated complex, the two cyclo\u00adpenta\u00addienyl rings adopt an eclipsed conformation. The anthracene tricycle is distorted towards a butterfly conformation.In a ferrocen\u00adyl\u2013anthracen-9(10 5H5)(C27H16NO)], designed and synthesized to explore a new electron-donor (D) and -acceptor (A) conjugated complex, the two cyclo\u00adpenta\u00addienyl rings adopt an eclipsed conformation. The anthracene tricycle is distorted towards a butterfly conformation and the mean planes of the outer benzene rings are inclined each to other at 22.7\u2005(3)\u00b0. In the crystal, mol\u00adecules are paired into inversion dimers via \u03c0\u2013\u03c0 inter\u00adactions. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions link further these dimers into one-dimensional columns along the b axis, with the ferrocenylethynyl arms arranged between the stacks to fill the voids.In the title compound, [Fe(C D) and -acceptor (A) mol\u00adecules have attracted much attention owing to their unique structures and various characteristic properties demonstrate guest-mol\u00adecule absorption and valence tautomerization etc. We have synthesized the title compound 1-(ferrocenylethyn\u00adyl)-10-(phenyl\u00adimino)\u00adanthracen-9(10H)-one [1-(Fc)AqPHI] and herein we report its crystal structure, determined by synchrotron radiation (SR) X-ray powder diffraction.Compounds containing a mixture of electron-donor 2Aq. Distances between the ring centroids cover the range from 4.09\u2005\u00c5 in 1,4 Fc2Aq down to 3.68\u2005\u00c5 in 1,2-(FcPh)2Aq. The smallest perpendicular distance for all the materials was close to 3.45\u2005\u00c5 . C\u2014H\u22ef\u03c0 inter\u00adactions are also found in 1,4-Fc2Aq, 1,5-Fc2Aq and 1,2-(FcPh)2Aq. Two kinds of C\u2014H\u22ef\u03c0 inter\u00adactions in 1,4-Fc2Aq connect the CP rings and the rings of the Aq groups of neighbouring mol\u00adecules. A C\u2014H\u22ef\u03c0 inter\u00adaction in 1,5-Fc2Aq links a CH\u2013 group from the Aq unit and a CP ring of Fc fragment. There are three C\u2014H\u22ef\u03c0 inter\u00adactions in 1,2-(FcPh)2Aq.In the reported examples compiled in the Cambridge Structural Database -one , ethynylferrocene , Pd(PPh3)2Cl2 (3.1\u2005mg), and CuI (5\u2005mg) were suspended in Et3N (15\u2005ml). After refluxing for 5\u2005h, Et3N was removed in vacuo, and the resultant residue was dissolved in CH2Cl2. The solution was washed with water (150\u2005ml), and dried over Na2SO4. After evaporation of the solvent, the crude product was purified with alumina column chromatography (activity II\u2013III) with a mixture of di\u00adchloro\u00admethane and hexane (1:2 v/v) as eluent. The third fraction was collected, and produced a red\u2013brown solid of the title compound . Very small single crystals unsuitable for conventional X-ray structure analysis were obtained by recrystallization from di\u00adchloro\u00admethane\u2013hexane. 1H NMR : \u03b4 8.1\u20138.5 , 7.0\u20137.9 , 6.80 , 4.1\u20134.8 . IR (KBr pellet): 2208 (\u03bd C=C/ cm\u22121), 1668 (\u03bd C=O/ cm\u22121), 1483 (\u03bd C=N/ cm\u22121). MALDI\u2013TOF\u2013MS: m/z = 490.1.Under a nitro\u00adgen atmosphere, 1-bromo-10-(phenyl\u00adimino)\u00adanthracen-9 standard powder sample was used for wavelength calibration. The calibrated wavelength was 0.80200\u2005(1)\u2005\u00c5. The powder profile was measured at 100\u2005K with 120\u2005min X-ray exposure time.The size of 1-(Fc)AqPHI crystals was small, less than 1\u2005\u00b5m. SR powder-diffraction techniques were employed for the structure determination. The powder crystallites were installed in a 0.4\u2005mm glass capillary. The X-ray powder diffraction data were measured using a large Debye\u2013Scherrer camera with an imaging-plate (IP) as a detector installed at SPring-8 BL02B2 was 63.2. The space group P21/n was assigned on the basis of systematic extinctions.Indexing was carried out using the program SP . Uiso for H atoms connected to the Aq and Ph parts were fixed at 0.05\u2005\u00c52. isoU for H atoms connected to the C25\u2013C29 and C30\u2013C34 CP rings were fixed at 0.09\u2005\u00c52 and 0.04\u2005\u00c52, respectively. A split-type pseudo-Voigt profile function global, I, 1FcAqPHI-100K_powder_data. DOI: 10.1107/S1600536814025252/cv5475Isup2.rtvRietveld powder data: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814025252/cv5475Isup3.molSupporting information file. DOI: 1034683CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two compounds are isotypic and the two-dimensional polymeric structure is based on centrosymmetric dinuclear bridged complex units. Within the layers, which lie parallel to (100), the coordinating water mol\u00adecule forms an O\u2014H\u22efO hydrogen bond to the single bridging carboxyl\u00adate O atom. 3-2-acetato]\u00addipotassium], [K2(C8H5Cl2O3)2(H2O)]n, and poly[\u03bc-aqua-bis\u00ad[\u03bc3-2-acetato]\u00addirubidium], [Rb2(C8H5Cl2O3)2(H2O)]n, respectively, have been determined and are described. The two compounds are isotypic and the polymeric structure is based on centrosymmetric dinuclear bridged complex units. The irregular six-coordination about the alkali cations comprises a bridging water mol\u00adecule lying on a twofold rotation axis, the phen\u00adoxy O-atom donor and a triple bridging carboxyl\u00adate O atom of the oxo\u00adacetate side chain of the 3,5-D ligand, and the second carb\u00adoxy\u00adate O-atom donor also bridging. The K\u2014O and Rb\u2014O bond-length ranges are 2.7238\u2005(15)\u20132.9459\u2005(14) and 2.832\u2005(2)\u20133.050\u2005(2)\u2005\u00c5, respectively, and the K\u22efK and Rb\u22efRb separations in the dinuclear units are 4.0214\u2005(7) and 4.1289\u2005(6)\u2005\u00c5, respectively. Within the layers which lie parallel to (100), the coordinating water mol\u00adecule forms an O\u2014H\u22efO hydrogen bond to the single bridging carboxyl\u00adate O atom.The two-dimensional coordination polymeric structures of the hydrated potassium and rubidium salts of acetic acid , namely, poly[\u03bc-aqua-bis\u00ad[\u03bc The comparative M\u2014O bond length range for the two metals (Tables 1O:O1)-chelate inter\u00adaction .The hydrated complexes (I)on Fig.\u00a02, three b Tables 1 and 2 \u25b8 M\u2014O complex network with the aromatic rings of the ligands peripherally located between the layers. Within the layers there are a number of short metal\u22efmetal contacts, the shortest being across an inversion centre [K\u22efKii = 4.0214\u2005(7)\u2005\u00c5 and Rb\u22efRbii = 4.1289\u2005(6)\u2005\u00c5], the longest being K\u22efKvi = 4.3327\u2005(5)\u2005\u00c5 and Rb\u22efRbvi = 4.5483\u2005(5)\u2005\u00c5 or Rb2CO3 (115\u2005mg) (for (II)] to a hot solution of acetic acid (220\u2005mg) in 10\u2005ml of 50% (v/v) ethanol/water. After heating for 5\u2005min, partial room temperature evaporation of the solutions gave in all two cases, colourless needles from which specimens were cleaved for the X-ray analyses.Compounds (I)aromatic = 0.95\u2005\u00c5 or C\u2014Hmethyl\u00adene = 0.99\u2005\u00c5] and were allowed to ride in the refinements, with Uiso(H) = 1.2Ueq(C). The water H-atom in both structures was located in a difference Fourier map and was allowed to ride in the refinements with an O\u2014H distance restraint of 0.90\u00b10.02\u2005\u00c5 and with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details for (I)10.1107/S2056989015016722/wm5206sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989015016722/wm5206Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989015016722/wm5206IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989015016722/wm5206Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015016722/wm5206IIsup5.cmlSupporting information file. DOI: 1422835, 1422834CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "O\u2014H\u22efO hydrogen bonds between water mol\u00adecules and ClO4 units lead to the formation of a three-dimensional network in the structures.The crystal structures of the tetra- and hexa\u00adhydrate phases of Ca6(ClO4)]2 dimers by sharing two ClO4 tetra\u00adhedra. The dimers are arranged in sheets parallel (001) and alternate with layers of non-coordinating ClO4 tetra\u00adhedra. O\u2014H\u22efO hydrogen bonds between the water mol\u00adecules as donor and ClO4 tetra\u00adhedra and water mol\u00adecules as acceptor groups lead to the formation of a three-dimensional network in the two structures. Ca(ClO4)2\u00b76H2O was refined as a two-component inversion twin, with an approximate twin component ratio of 1:1 in each of the two structures.The title compounds, calcium perchlorate tetra\u00adhydrate and calcium perchlorate hexa\u00adhydrate, were crystallized at low temperatures according to the solid\u2013liquid phase diagram. The structure of the tetra\u00adhydrate consists of one Ca The resulting coordination polyhedron is a distorted square anti-prism . The Ca\u2014O bond lengths involving the water mol\u00adecules range from 2.3284\u2005(17) to 2.4153\u2005(16)\u2005\u00c5 and are considerably shorter than the Ca\u2014O bond lengths involving the perchlorate O atoms [2.5417\u2005(16) to 2.5695\u2005(17)\u2005\u00c5].The Casm Fig.\u00a01b. The C2+ cations in Ca(ClO4)\u00b76H2O are each coordinated by six water mol\u00adecules and two perchlorate tetra\u00adhedra . Nevertheless, according to the bond-valence model 6(ClO4)]2 dimers oriented in layers parallel to (001).The two different Cara Fig.\u00a02. Again, 4)\u00b74H2O are shared between two adjacent Ca2+ ions, forming chains extending parallel to 2 sheets along [001] 2\u00b74H2O phases, see: Robertson & Bish (2010M = Mg); Hennings et al. (2014M(ClO4)2\u00b76H2O phases, see: Ghosh et al. ; Ghosh & Ray 2\u00b74H2O was crystallized from an aqueous solution of 62.96 wt% Ca(ClO4)2 at 273\u2005K after one day and Ca(ClO4)2\u00b76H2O from an aqueous solution of 57.55 wt% Ca(ClO4)2 at 238\u2005K after one week. For the preparation of these aqueous solutions, Ca(ClO4)2\u00b74H2O was used. The Ca2+ content was analysed via complexometric titration with EDTA. The crystals remain stable in the saturated aqueous solution over at least four weeks.Ca2\u00b74H2O, distance restraints were applied for all water mol\u00adecules, with O\u2014H and H\u2014H distance restraints of 0.82\u2005(1) and 1.32\u2005(1)\u2005\u00c5, respectively. For Ca(ClO4)2\u00b76H2O, Uiso values were set at 1.2Ueq(O) using a riding-model approximation. Distance restraints were applied for that structure for all water mol\u00adecules, with O\u2014H and H\u2014H distance restraints of 0.84\u2005(2) and 1.4\u2005(2)\u2005\u00c5, respectively. Ca(ClO4)2\u00b76H2O was refined as a two-component inversion twin, with an approximate twin component ratio of 1:1.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S1600536814024532/wm5079sup1.cifCrystal structure: contains datablock(s) CaClO4_4H2O_200K, CaClO4_6H2O_180K. DOI: 10.1107/S1600536814024532/wm5079CaClO4_4H2O_200Ksup2.hklStructure factors: contains datablock(s) CaClO4_4H2O_200K. DOI: 10.1107/S1600536814024532/wm5079CaClO4_6H2O_180Ksup3.hklStructure factors: contains datablock(s) CaClO4_6H2O_180K. DOI: Click here for additional data file.10.1107/S1600536814024532/wm5079CaClO4_4H2O_200Ksup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S1600536814024532/wm5079CaClO4_6H2O_180Ksup5.cmlSupporting information file. DOI: 1033323, 1033324CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Crystal structures of morpholinium hydrogen bromanilate have been determined at 130, 145 and 180\u2005K. The asymmetric unit comprises one morpholinium cation and two halves of crystallographically independent bromanilate monoanions. The conformations of the two independent bromanilate anions are different from each other with respect to the O\u2014H orientation. The two different anions are linked alternately into a chain though a short O\u2014H\u22efO hydrogen bond, in which the H atom is disordered over two positions. 4H10NO+\u00b7C6HBr2O4\u2212, were determined at three temperatures, viz. 130, 145 and 180\u2005K. The asymmetric unit comprises one morpholinium cation and two halves of crystallographically independent bromanilate monoanions, which are located on inversion centres. The conformations of the two independent bromanilate anions are different from each other with respect to the O\u2014H orientation. In the crystal, the two different anions are linked alternately into a chain along [211] through a short O\u2014H\u22efO hydrogen bond, in which the H atom is disordered over two positions. The refined site-occupancy ratios, which are almost constant in the temperature range studied, are 0.49\u2005(3):0.51\u2005(3), 0.52\u2005(3):0.48\u2005(3) and 0.50\u2005(3):0.50\u2005(3), respectively, at 130, 145 and 180\u2005K, and no significant difference in the mol\u00adecular geometry and the mol\u00adecular packing is observed at the three temperatures. The morpholinium cation links adjacent chains of anions via N\u2014H\u22efO hydrogen bonds, forming a sheet structure parallel to (-111).Crystal structures of the title compound , C The anomalous behavior was ascribed to a drastic temperature variation of the disordered O\u2014H\u22efO hydrogen bond, as revealed by multi-temperature X-ray diffraction \u2005\u00c5 at 114\u2005K to 1.2951\u2005(10)\u2005\u00c5 at 180\u2005K; C6\u2014O4: from 1.290\u2005(10)\u2005\u00c5 at 114\u2005K to 1.2946\u2005(10) at 180\u2005K], which corresponds to population changes of the two disordered proton sites in the hydrogen bond (Tobu i: 3.1698\u2005(13)\u2005\u00c5 (130\u2005K), 3.1725\u2005(13)\u2005\u00c5 (145\u2005K) and 3.1763\u2005(13)\u2005\u00c5 (180\u2005K); Br2\u22efC1i: 3.2673\u2005(15)\u2005\u00c5 (130\u2005K), 3.2716\u2005(15)\u2005\u00c5 (145\u2005K) and 3.2808\u2005(15)\u2005\u00c5 (180\u2005K); symmetry code: (i) x- 1, y\u00a0\u2212\u00a01, z] are observed. A weak C\u2014H\u22efBr inter\u00adaction is also observed between the sheets.In the crystal, the two independent bromanilate anions with different conformations are linked alternately by short O\u2014H\u22efO hydrogen bonds Tables 1 and 3 \u25b8,A form of bromanilic acid (200\u2005mg) with morpholine (60\u2005mg) at room temperature.Uiso(H) = 1.2Ueq(C). The N-bound H atom was located in a difference Fourier map and refined freely [refined N\u2014H = 0.85\u2005(3)\u20130.89\u2005(3)\u2005\u00c5]. Two disordered positions of the H atom in the O\u2014H\u22efO hydrogen bond were located in a difference Fourier map. Since site occupancy factors and isotropic displacement parameters are correlated and bonding effects also make the site-occupancy factors less reliable, the positional parameters and the occupancies of the H atom were refined, with Uiso(H) = 1.5Ueq(O), and with distance restraints of O\u2014H = 0.84\u2005(2)\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989015017272/lh5788sup1.cifCrystal structure: contains datablock(s) General, 1, 2, 3. DOI: 10.1107/S2056989015017272/lh57881sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989015017272/lh57882sup3.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S2056989015017272/lh57883sup4.hklStructure factors: contains datablock(s) 3. DOI: Click here for additional data file.10.1107/S2056989015017272/lh57881sup5.cmlSupporting information file. DOI: 1424714, 1424713, 1424712CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal packing features \u03c0\u2013\u03c0 stacking inter\u00adactions between tpy\u2013tpy ligands.In this first crystal structure of an Ru complex with 6\u2032,6\"-bis\u00ad(pyridin-2-yl)-2,2\u2032:4\u2032,4\":2\",2\"\u2019-quaterpyridine, a \u2018half\u2019 of the ligand (one of the two terpyridyl units) is N^N^N 10H8N2)(C30H20N6)]PF6, which contains the bidentate ligand 2,2\u2032-bi\u00adpyridine (bpy) and the tridendate ligand 6\u2032,6\u2032\u2032-bis\u00ad(pyridin-2-yl)-2,2\u2032:4\u2032,4\u2032\u2032:2\u2032\u2032,2\u2032\u2032\u2032-quaterpyridine (tpy\u2013tpy). The [RuCl(bpy)(tpy\u2013tpy)]+ monocation has a distorted octa\u00adhedral geometry at the central RuII ion due to the restricted bite angle [159.32\u2005(16)\u00b0] of the tridendate ligand. The Ru-bound tpy and bpy moieties are nearly planar and essentially perpendicular to each other with a dihedral angle of 89.78\u2005(11)\u00b0 between the least-squares planes. The lengths of the two Ru\u2014N bonds for bpy are 2.028\u2005(4) and 2.075\u2005(4)\u2005\u00c5, with the shorter bond being opposite to Ru\u2014Cl. For tpy\u2013tpy, the mean Ru\u2014N distance involving the outer N atoms trans to each other is 2.053\u2005(8)\u2005\u00c5, whereas the length of the much shorter bond involving the central N atom is 1.936\u2005(4)\u2005\u00c5. The Ru\u2014Cl distance is 2.3982\u2005(16)\u2005\u00c5. The free uncoordinated moiety of tpy\u2013tpy adopts a trans,trans conformation about the inter\u00adannular C\u2014C bonds, with adjacent pyridyl rings being only approximately coplanar. The crystal packing shows significant \u03c0\u2013\u03c0 stacking inter\u00adactions based on tpy\u2013tpy. The crystal structure reported here is the first for a tpy\u2013tpy complex of ruthenium. We report the structural characterization of \u00adchlorido\u00adruthenium(II) hexa\u00adfluorido\u00adphosphate, [RuCl(C In this and other related photocatalysts containing the {(tpy)Ru(bpy)(L)} moiety (L = H2O or Cl\u2212), the aqua species is typically formed by easy ligand substitution from its chlorido precursor in water Ru(tpy\u2013tpy)Ru(bpy)(Cl)]3+ crystallizes in the triclinic \u00b0 in agreement with those found in similar chlorido RuII-bpy complexes \u00b0]. The RuII atom and atoms N2, N4, N5, and Cl1 form an equatorial plane with a maximum deviation of 0.032\u2005(4)\u2005\u00c5. The Ru-bound tpy moiety and bpy are approximately planar and their mean planes are essentially perpendicular to each other with a dihedral angle of 89.78\u2005(11)\u00b0 between planes. For the tridentate ligand, the mean Ru\u2014N distance involving the outer N1 and N3 atoms trans to each other is 2.053\u2005(8)\u2005\u00c5, whereas the bond distance involving the central N2 is much shorter [1.936\u2005(4)\u2005\u00c5] as a result of the structural constraint imposed by these mer-arranged ligands and plane\u2013plane dihedral angles (\u03b1) are respectively: 3.723\u2005(3)\u2005\u00c5 and 2.8\u2005(2)\u00b0 for \u22ef ; 3.812\u2005(4)\u2005\u00c5 and 3.2\u2005(2)\u00b0 for \u22ef ; 3.826\u2005(4)\u2005\u00c5 and 5.6\u2005(3)\u00b0 for \u22ef ; and 3.630\u2005(4)\u2005\u00c5 and 15.5\u2005(3)\u00b0 for \u22ef . In all these \u03c0\u2013\u03c0 stacking inter\u00adactions, the slip angles from the parallel displacement are smaller than 30\u00b0.The intra\u00admolecular Cl\u22efH contact of 2.70\u2005\u00c5 involving the hydrogen of the nearest C atom at bpy (H25) is similar to that observed earlier for complexes containing the {RuCl(bpy)} moiety -2,2\u2032:4\u2032,4\u2032\u2032:2\u2032\u2032,2\u2032\u2032\u2032-quaterpyridine. Other than one structure for the metal-free ligand itself (DMSO)2Cl2 into a solution of the tpy\u2013tpy ligand at reflux. The reaction solution was refluxed for another 2.5\u2005h and then cooled down to room temperature. After evaporation of the solvent on a rotavap, water was added to dissolve the solid and excess NH4PF6 was added to form the precipitate, which was filtered off and dried under vacuum. Further purification was performed by column chromatography using alumina and a mixture of aceto\u00adnitrile/toluene (1:2) as the eluant. The product was collected from the first band. The solvent was evaporated and the dark-red solid was collected and dried under vacuum (yield: 30%). Analysis calculated for C40H28N8F6PClRu: C, 53.25; H, 3.13; N, 12.42. Found: C, 52.71; H, 3.12; N, 11.86. Single crystals for X-ray structural analysis were grown by slow diffusion of diethyl ether into aceto\u00adnitrile solutions of the complexes in long thin tubes.Compound + was also characterized in MeCN solutions by other techniques. Mass spectra (ESI\u2013MS: m/z 757) are in agreement with the formulation for the cation, i.e. [1(-PF6)]+ . 1H-NMR : \u03b4 10.27\u201310.26 , 9.07 , 8.89 , 8.73\u20136.95 . Electrochemical measurements by cyclic voltammetry gave a redox potential of 0.83\u2005V vs SCE for the reversible RuII/RuIII couple. This potential is anodically shifted by only 20\u2005mV relative to the [Ru(Cl)(bpy)(tpy)]+ complex (bpy)(tpy\u2013tpy)]Uiso(H) = 1.2Ueq(C). Each atom in the anion was modeled in two positions, with site occupancies tied to 1.0. A total of 48 temperature-factor restraints were used to force convergence. The SQUEEZE routine in PLATON I. DOI: 10.1107/S2056989015014632/pk2553Isup2.hklStructure factors: contains datablock(s) I. DOI: 1416756CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked by N\u2013H\u22efN 11H9N5OS2, a 1-thio\u00adphen-2-yl\u00admethyl\u00adene\u00adamino\u00adpyrimidine derivative, displays an essentially planar C\u2014NH2 group. The conformation across the N=C bond linking the pyrimidine and thienyl groups is E. The pyrimidine and thienyl ring systems subtend an inter-planar angle of 42.72\u2005(5)\u00b0. In the crystal, mol\u00adecules are linked by N\u2013H\u22efNnitrile and N\u2013H\u22efO=C hydrogen bonds, forming chains parallel to the b axis.The title compound, C Many drugs, such as 5-fluoro\u00aduracil, containing a pyrimidine moiety have been developed and used as anti\u00adcancer agents. It is difficult to find a general method for the introduction of specific substituents into the pyrimidine nucleus directly, and thus many synthetic methods have been developed for the construction of pyrimidine rings bearing potential functional groups \u2005\u00c5. Both ring systems are, as expected, planar (r.m.s. deviations are 0.017\u2005\u00c5 for the pyrimidine and 0.001\u2005\u00c5 for the thienyl ring). Atom N2 lies 0.189\u2005(2)\u2005\u00c5 out of the pyrimidine plane; all other immediate substituent atoms lie effectively in the ring plane. Carbon atom C7 of the thio\u00admethyl group is rotated slightly out of the ring plane, with torsion angle N3\u2014C4\u2014S1\u2014C7 being \u22126.30\u2005(10)\u00b0. The inter-planar angle between the rings is 42.72\u2005(5)\u00b0; the relative orientation is influenced by the torsion angles C6\u2014N1\u2014N2\u2014C10 = \u221251.78\u2005(13), N1\u2014N2\u2014C10\u2014C11 = 174.68\u2005(9) and N2\u2014C10\u2014C11\u2014S12 = 5.22\u2005(15)\u00b0. The NH2 group is planar; the nitro\u00adgen atom lies only 0.048\u2005(9)\u2005\u00c5 out of the plane of its substituents. The intra\u00admolecular contact H041\u22efN2 = 2.22\u2005(2)\u2005\u00c5 may be construed as a hydrogen bond, although the angle at the H atom is necessarily narrow at 108.4\u2005(14) \u00b0 \u2005\u00c5 and H042\u22efO1 2.14\u2005(2)\u2005\u00c5 Table\u00a01.A search of the Cambridge Structural Database was added to a stirred solution of 1-cyano\u00adacetyl-4-thio\u00adphene\u00admethyl\u00adidenesemicarbazide (0.01\u2005mol) in dry dioxane (50\u2005ml), containing potassium hydroxide (0.01\u2005mol), at room temperature. The solution was stirred overnight at room temperature, after which a colourless solid product was collected by filtration and crystallized from ethanol (m.p. 541\u2013542\u2005K), giving colourless block-like crystals.Dimethyl Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms. The methyl group was refined as an idealized rigid group, allowed to rotate but not to tip.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901501885X/su5217sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901501885X/su5217Isup2.hklStructure factors: contains datablock(s) I. DOI: 1430030CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dihedral angle between the mean plane of the benzene and piperidine rings is 89.03\u2005(3)\u00b0. In the crystal structure, inversion dimers are formed via N\u2014H\u22efCl hydrogen-bond inter\u00adactions, resulting in chains parallel to the [001] direction. The benzene rings within the chains show \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid-to-centroid distances of 3.801\u2005(3)\u2005\u00c5] and neighbouring chains inter\u00adact via N\u2014H\u22efO hydrogen bonds.In the title complex, [PtCl Then, in the reaction solution, the cis complex converts into the trans complex and the thermodynamics of this conversion are currently under investigation by us.To explain this we suppose that et al. \u00b0]. The piperidine ring adopts the usual chair conformation, with the N2\u2014Pt1 bond in the equatorial position. The piperidine ring is oriented nearly perpendicular to the coordination plane of the PtII atom, thereby reducing the van der Waals repulsion; the dihedral angle between the least-squares mean planes through the piperdine ring and the four atoms coordinated to the Pt atom is 89.6\u2005(2)\u00b0. One short intra\u00admolecular contact is observed, i.e. H7B\u22efCl8 = 2.83\u2005\u00c5. The mean planes through the piperidine ring and the benzene ring make a dihedral angle of 89.0\u2005(3)\u00b0. The dihedral angle between the mean planes of the nitro substituent and the benzene ring is 16.6\u2005(3)\u00b0.The mol\u00adecular structure of the title compound is illustrated in Fig.\u00a01via N\u2014H\u22efCl inter\u00adactions between the aniline N atom and both Cl atoms, resulting in chains of mol\u00adecules along the [001] direction \u2005\u00c5; Cg is the centroid of the C11\u2013C16 ring; symmetry code: (iv) \u2212x, y, \u2212z\u00a0+\u00a0via N\u2014H\u22efO hydrogen bonds between the piperidine N atom and a nitro O atom , with the remaining 239 structures showing a trans coordination. There is no difference in the Pt\u2014Cl distances between both configurations. The average Pt\u2014Cl distances are 2.300\u2005(15) and 2.299\u2005(12)\u2005\u00c5 for the cis and trans arrangements, respectively, and correspond to the observed distances of 2.3039\u2005(11) and 2.2917\u2005(12)\u2005\u00c5 for Pt1\u2014Cl8 and Pt1\u2014Cl9, respectively.A search of the Cambridge Structural Database ] , prepared according to the synthetic procedure of Da et al. (2001p-nitro\u00adaniline (1\u2005mmol) in ethanol (10\u2005ml) was added gradually while stirring at 413\u2013318\u2005K. After 1\u2005h, a brown powder appeared and the reaction mixture was then stirred further for 24\u2005h until all the precipitate was completely dissolved. The solvent was removed in vacuo to give a brown\u2013yellow product. The product was washed consecutively with a 0.1 M HCl solution (2 \u00d7 2\u2005ml), warm water (2 \u00d7 2\u2005ml) and diethyl ether (2 \u00d7 2\u2005ml). The yield was 80%. Single crystals suitable for X-ray determination were obtained by slow evaporation within 12\u2005h from an acetone solution at room temperature. IR : 3199, 3113 (\u03bdNH); 3070, 2927, 2862 (\u03bdCH); 1596, 1525, 1479 (\u03bdC=C arom); 1342, 1325 (\u03bdNO); 1H NMR : \u03b4 8.21 , 7.47 , 5.49 , 3.66 , 3.26 , 2.99 = 13.0\u2005Hz, C510HNH), 1.69\u20131.43 . 13C{1H} NMR : \u03b4\u00a0149.6, 125.1, 124.2 (O2N6CH4NH2), 54.0, 27.2, 24.3 (5CH10NH).The starting complex K[PtCl al. 2001 with sliUiso(H) values assigned as 1.2Ueq of the parent atoms, with C\u2014H distances of 0.95 (aromatic) and 0.99\u2005\u00c5 (methyl\u00adene), and N\u2014H distances of 0.93 (NH) and 0.92\u2005\u00c5 (NH2).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015009196/rz5159sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015009196/rz5159Isup2.hklStructure factors: contains datablock(s) I. DOI: 1400786CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-(ar\u00adyl)-2,2,2-tri\u00adbromo\u00adacetamides show different weak inter\u00adactions in their crystal structures.Three N-ar\u00adyl-2,2,2-tri\u00adbromo\u00adacetamides, namely, 2,2,2-tri\u00adbromo-N-(2-fluoro\u00adphen\u00adyl)\u00adacetamide, C8H5Br3FNO, (I), 2,2,2-tri\u00adbromo-N-[3-(tri\u00adfluoro\u00admethyl)\u00adphen\u00adyl]\u00adacetamide, C9H5Br3F3NO, (II) and 2,2,2-tri\u00adbromo-N-(4-fluoro\u00adphen\u00adyl)\u00adacetamide, C8H5Br3FNO, (III) were synthesized and their crystal structures were analysed. In the crystal structure of (I), C\u2014Br\u22ef\u03c0ar\u00adyl inter\u00adactions connect the mol\u00adecules into dimers, which in turn are connected via Br\u22efBr contacts [3.6519\u2005(12)\u2005\u00c5], leading to the formation of a one-dimensional ladder-type architecture. The crystal structure of (II) features chains linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. Two such chains are inter\u00adlinked to form ribbons through Br\u22efBr [3.6589\u2005(1)\u2005\u00c5] and Br\u22efF [3.0290\u2005(1)\u2005\u00c5] inter\u00adactions. C\u2014Br\u22ef\u03c0ar\u00adyl and C\u2014F\u22ef\u03c0ar\u00adyl inter\u00adactions between the ribbons extend the supra\u00admolecular architecture of (II) from one dimension to two. In (III), the mol\u00adecules are connected into R22(8) dimers via pairs of C\u2014H\u22efF inter\u00adactions and these dimers form ribbons through Br\u22efBr [3.5253\u2005(1)\u2005\u00c5] contacts. The ribbons are further inter\u00adlinked into columns via C\u2014Br\u22efO=C contacts, forming a two-dimensional architecture.Three N-Ar\u00adyl-halo\u00adamides show a broad spectrum of pharmacological properties, including anti\u00adbacterial \u00adacetamide, (I)N-[3-(tri\u00adfluoro\u00admethyl)\u00adphen\u00adyl]\u00adacetamide, (II)N-(4-fluoro\u00adphen\u00adyl)\u00adacetamide, (III)The mol\u00adecular structures of (I)syn to the 2-fluoro substituent in the benzene ring, similar to that observed in the crystal structures of other ortho substituted compounds (see database survey). Contrast to the above, in (II)anti to the 3-CF3 substituent.In (I)In (I)The dihedral angle between the benzene ring and the C1\u2013N1\u2013C7(O)\u2013C8 segment in (II)The dihedral angle between the benzene ring and the C1\u2013N1\u2013C7(O)\u2013C8 segment in (III)ar\u00adyl inter\u00adactions \u00b0. The crystal structure of (I)In the crystal structure of (I)s Table\u00a04 connect re Fig.\u00a04. The Br1ar\u00adyl and C9\u2014F2\u22ef\u03c0ar\u00adyl inter\u00adactions between the ribbons extend the supra\u00admolecular architecture of (II)et al., 2012cis contact.The crystal structure of (II)via pairs of C3\u2014H3\u22efF1 inter\u00adactions ns Fig.\u00a07, formings Table\u00a06, formings Table\u00a06. The pacN-ar\u00adyl-2,2,2-tri\u00adbromo\u00adacetamides, namely, 2,2,2-tri\u00adbromo-N-phen\u00adyl\u00adacetamide, 2,2,2-tri\u00adbromo-N-(2/3/4-chloro\u00adphen\u00adyl)\u00adacetamides and 2,2,2-tri\u00adbromo-N-(2/3/4-methyl\u00adphen\u00adyl)\u00adacetamides have been previously reported. Comparison of the crystal systems of these series of compounds show that all the chloro-substituted compounds crystallize in the ortho\u00adrhom\u00adbic crystal system, while the methyl-substituted compounds crystallize in the monoclinic system (Table\u00a07i.e. (I)Seven m Table\u00a07. Howeversyn to the 2-fluoro substituent in the benzene ring, similar to that observed in the crystal structures of 2,2,2-tri\u00adbromo-N-(2-chloro\u00adphen\u00adyl)\u00adacetamide (Ia) \u00adacetamide (Ib) \u00adacetamide (Ia) \u00adacetamide (Ib) intra\u00admolecular hydrogen bonds. Further, compounds (I)In (I)i.e. unsubstituted) compound is closer to those of chloro-substituted ones, thus the order is F < Cl(=H) < CH3.A comparison of the dihedral angle between the benzene ring and the C1\u2013N1\u2013C7(O)\u2013C8 segment in all of the compounds shows that the dihedral angles in the fluoro-substituted compounds are smaller than those observed in chloro-substituted ones, which in turn have smaller values than the methyl-substituted tri\u00adbromo\u00adacetamides Table\u00a07. The diha)] reported in the literature feature strong N\u2014H\u22efO hydrogen bonds leading into C(4) chains forming a one-dimensional architecture. Compound (Ia) (2-chloro derivative) does not exhibit any conventional inter\u00admolecular inter\u00adactions and therefore exhibits a zero-dimensional supra\u00admolecular architecture. However, the packing of mol\u00adecules in the three structures reported here are very different and are controlled by inter\u00adactions mainly involving the halogen atoms.The crystal structures of all of the seven compounds [except = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0810.1107/S2056989015015248/hb7472sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989015015248/hb7472Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989015015248/hb7472IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989015015248/hb7472IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 1064065, 1419188, 1419189CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In membrane proteins, proline-mediated helix kinks are indispensable for the tight packing of transmembrane (TM) helices. However, kinks invariably affect numerous interhelical interactions, questioning the acceptance of proline substitutions and evolutionary origin of kinks. Here, we present the structural and thermodynamic basis of proline-induced integrin \u03b1IIb\u03b23 TM complex stabilization to understand the introduction of proline kinks in membrane proteins. In phospholipid bicelles, the A711P substitution in the center of the \u03b23 TM helix changes the direction of adjacent helix segments to form a 35\u2009\u00b1\u20092\u00b0 angle and predominantly repacks the segment in the inner membrane leaflet due to a swivel movement. This swivel repacks hydrophobic and electrostatic interhelical contacts within intracellular lipids, resulting in an overall TM complex stabilization of \u22120.82\u2009\u00b1\u20090.01\u2009kcal/mol. Thus, proline substitutions can directly stabilize membrane proteins and such substitutions are proposed to follow the structural template of integrin \u03b1IIb\u03b23(A711P). In the evolution of globular proteins, structural complexity and functionality can be increased by combining independently folding protein domains13456Indirect support for this hypothesis is abundant. Inspection of membrane protein structures reveals that helix kinks are frequently centered around proline residues 4689. Eve468TM, of \u22124.84\u2009\u00b1\u20090.01\u2009kcal/mol and \u03b23(W715), which are located at the N- and C-helix termini, and by detecting NOEs to the aromatic ring of \u03b1IIb(F993) in fractionally deuterated samples (21To understand the basis of \u03b23(A711P), we determined the structure of the \u03b1IIb\u03b23(A711P) TM complex in isotropic phospholipid bicelles by multidimensional heteronuclear NMR spectroscopy. In the structure determination of the wild-type \u03b1IIb\u03b23 TM complex, we had used selectively methyl-labeled protein and deuterated lipids to obtain interhelical NOE distance restraintsH\u03b1 pairs , albeit samples . Membranamples 21.15N chemical shift differences between \u03b1IIb when complexed with either \u03b23 or \u03b23(A711P) illustrated that structural changes predominantly took place for IMC residues and residues that pack near the mutation site \u2212 (\u0394G\u00b0\u03b1IIb\u03b23(A711P),mutant \u2212\u0394G\u00b0\u03b1IIb\u03b23(A711P)) was quantified to compare the disturbance created by a mutation relative to its respective \u03b1IIb\u03b23 and \u03b1IIb\u03b23(A711P) reference structure. In accordance with largely invariant OMC interactions, \u0394\u0394G\u00b0,\u2032 was small for \u03b1IIb(G972A) with 0.16\u2009\u00b1\u20090.03\u2009kcal/mol and \u03b1IIb\u03b23 TM complexes. Specifically, \u0394\u0394G\u00b0,\u2032\u2009=\u2009, prolines in membrane protein structures are frequently encountered near the center of TM helices422TM must be balanced with the affinity of intra- and extracellular receptor agonists and with the stability of the inactive versus the active ectodomains15\u03a4\u039c came at the expense of \u03b1IIb(R995)-\u03b23(D723) destabilization. This interaction is disrupted during talin-mediated integrin activationThe alternative to maintaining interhelical contacts near the proline kink would be to preserve interactions at the TM helix termini. When inspecting this possibility for \u03b1IIb\u03b23(A711P), it is apparent that mostly \u03b1IIb(R995)-\u03b23(D723) benefits whereas packing on \u03b23(G708) and \u03b1IIb(G976) would be less intimate . This mo7-glucose, 99% 15ND4Cl and 99% D2O. A fractionally deuterated 2H/13C/15N-\u03b1IIb(A963C)\u2013\u03b23(G690C/A711P) sample was prepared by growing E. coli cells in 60% D2O using protonated precursors. Freeze-dried peptide was reconstituted in 320 \u03bcL of 350\u2009mM 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC), 105\u2009mM 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 6% D2O, 0.02% w/v NaN3 buffered by either 25\u2009mM NaH2PO4/Na2HPO4, pH 7.4 or 25\u2009mM HEPES\u00b7NaOH, pH 7.4 for a final concentration of 0.8\u2009mM and bicelle q-factor of 0.3.The disulfide-linked \u03b1IIb(A963C)\u2013\u03b23(G690C/A711P) dimer was prepared applying published protocols1HN, 15N, 13C\u03b1, 13C\u03b2, and 13C\u2032 assignment of the \u03b1IIb\u03b23 TM complex and the \u03b23(A711P/K716A) TM segment152H/13C/15N-\u03b1IIb(A963C)\u20132H/13C/15N-\u03b23(G690C/A711P) were achieved employing HNCA, HNCO, HNCACB and NOESY-TROSY experiments. 15N-edited NOESY-TROSY experiments using 2H/15N-\u03b1IIb(A963C)\u2013\u03b23(G690C/A711P) or \u03b1IIb(A963C)\u20132H/15N-\u03b23(G690C/A711P) dimers were acquired with mixing times of 120, 150 and 175\u2009ms. Using [60% 2H]/13C/15N-\u03b1IIb(A963C)\u2013\u03b23(G690C/A711P), an aromatic 13C-edited NOESY-HSQC experiment (mixing time 150\u2009ms) was recorded. Sidechain assignments started again from the \u03b1IIb\u03b23 TM complex and were similar to the aforementioned NOESY spectra. In a general case, NOESY experiments for 2H/14N-\u03b1\u20131H/15-\u03b2 and 1H/15N-\u03b1\u20132H/14-\u03b2 can establish sidechain assignments in combination with standard experiments. Sidechain and NOE assignments were carried out manually using the program CARA. H-N residual dipolar couplings (RDC) were measured twice in compressed polyacrylamide gels using 2H/15N-\u03b1IIb(A963C)\u20132H/15N-\u03b23(G690C/A711P) dimerStarting from the 15N, 13C\u03b1, 13C\u03b2, and 13C\u2032 chemical shift patterns\u03b1-C\u2032, N-C\u2032 RDCs measured for these monomers to further restrict the individual \u03b1IIb and \u03b23(A711P) backbone conformations. An employed torsion angle potential of mean force1 angles detected in the monomeric \u03b1IIb and \u03b23(A711P/K716A) TM segments, which mostly corresponded to their default values. Moreover, the sidechains of \u03b1IIb(Phe992) and \u03b23(Lys716) were adjusted to snorkel. Aside from standard force field terms for covalent geometry and nonbonded contacts , dihedral angle restraints were implemented using quadratic square-well potentials. In addition, a backbone-backbone hydrogen-bonding potential was employed1D). The final values for the force constants of the different terms in the simulated annealing target function were as previously describedStructure calculations were carried out by simulated annealing, starting at 3000\u2009K using the program XPLOR-NIHsn-glycero-3-phosphocholine (DHPC), 17\u2009mM 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 25\u2009mM NaH2PO4/Na2HPO4 pH 7.4 at 28\u2009\u00b0C. Prior to data analysis, the measurements were corrected for the heat of dilutions of the \u03b1IIb and \u03b23 peptides. The \u03b1IIb\u03b23 complex stoichiometry was fixed at 1:1XY were calculated from the measured heat changes, \u03b4Hi, as described previouslyITC measurements of the peptides listed in How to cite this article: Schmidt, T. et al. Structural and thermodynamic basis of proline-induced transmembrane complex stabilization. Sci. Rep.6, 29809; doi: 10.1038/srep29809 (2016)."} +{"text": "E conformation about the azo\u00adbenzene linkage and the benzene rings are almost coplanar to one another [dihedral angle = 1.36\u2005(7)\u00b0]. In the crystal, a combination of O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions leads to the formation of slabs parallel to (001).The title compound has an 16H14N2O3, has an E conformation about the azo\u00adbenzene [\u2014N=N\u2013 = 1.2481\u2005(16)\u2005\u00c5] linkage. The benzene rings are almost coplanar [dihedral angle = 1.36\u2005(7)\u00b0]. The O atoms of the carb\u00adoxy\u00adlic acid group are disordered over two sets of sites and were refined with an occupancy ratio of 0.5:0.5. The two disordered components of the carb\u00adoxy\u00adlic acid group make dihedral angles of 1.5\u2005(14) and 3.8\u2005(12)\u00b0 with the benzene ring to which they are attached. In the crystal, mol\u00adecules are linked via pairs of O\u2014H\u22efO hydrogen bonds, forming inversion dimers. The dimers are connected via C\u2014H\u22efO hydrogen bonds, forming ribbons lying parallel to [120]. These ribbons are linked via C\u2014H\u22ef\u03c0 inter\u00adactions, forming slabs parallel to (001).The title compound, C Hence, liquid crystallinity may be induced by the formation of hydrogen-bonded dimers. A number of liquid crystal (LC) systems containing hydrogen bonds that function between identical mol\u00adecules have been reported linkage, the length of the N1\u2014N2 bond is 1.2481\u2005(16)\u2005\u00c5 and the torsion angle for the azo unit (C7\u2014N1=N2\u2014C10) is 179.99\u2005(10)\u00b0, which is comparable with the values of ca \u00b1180\u00b0 observed in 4,4-azinodi\u00adbenzoic acid -ethyl-4-{[4-(deca\u00adnoxl\u00adoxy)phen\u00adyl]diazenly} benzoate -2-[4-(but-3-en-1-yl\u00adoxy)phen\u00adyl]-diazen-1-yl}benzoic acid, , as shown in Fig.\u00a03In the crystal, mol\u00adecules are linked s Table\u00a01. The dimet al., 2012. The terminal double bonds-containing azo\u00adbenzene compound was hydrolysed under basic conditions to yield the title compound. Red plate-like crystals were obtained by crystallization from an ethanol\u2013ethyl acetate mixture (1:1); m.p. 494\u2005K. 1H NMR (CDCl3): \u03b4 8.18 , 7.94 , 7.93 , 7.05 , 6.04 , 5.45 , 5.31 , 4.60 .The title compound was synthesized by a literature procedure (Rahman Uiso(H) = 1.5 Ueq(O). The C-bound H atoms were positioned geometrically and refined using a riding model: C\u2014H = 0.93\u20130.97\u2005\u00c5 with Uiso(H) = 1.2Ueq(C). Two outlier reflections, 341 and 309, were omitted from the refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814023745/su2790sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814023745/su2790Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814023745/su2790Isup3.cmlSupporting information file. DOI: 1031374CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 contacts, resulting in a three-dimensional supra\u00admolecular structure.The title spiro-compound bears 17H18O3, the two non-spiro C atoms of the cyclo\u00adpropane ring bear a formyl and a phenyl substituent which are trans-oriented. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 contacts resulting in a three-dimensional supra\u00admolecular structure.In the title compound, C The angles in the three-membered ring range from 58.80\u2005(13)\u00b0 (C2\u2014C4\u2014C3) to 61.67\u2005(13)\u00b0 (C3\u2014C2\u2014C4) being close to the ideal value of 60\u00b0 for such a ring. The six-membered ring containing the spiro atom C4 and ring atoms C5\u2013C9 adopts a chair conformation with a puckering amplitude Q of 0.491\u2005(2)\u2005\u00c5 and \u03b8 = 16.8\u2005(2)\u00b0, which indicates a slight deviation from an ideal chair conformation with \u03b8 = 0\u00b0. The plane of the central cyclo\u00adpropane ring forms dihedral angles of 66.89\u2005(16) and 89.33\u2005(16)\u00b0, respectively, with the plane of the phenyl ring and the mean plane of the cyclo\u00adhexane ring [maximum deviation from this plane is 0.272\u2005(2)\u2005\u00c5 for atom C7]. The latter two planes form a dihedral angle of 64.15\u2005(10)\u00b0. The plane of the formyl group, consisting of atoms C1, H1 and O1, is almost normal to the cyclo\u00adpropane ring with a dihedral angle of 81.3\u2005(3)\u00b0.The mol\u00adecular structure of the title compound is depicted in Fig.\u00a01B\u22efO2 contacts (red dotted lines) which are further linked to double strands along the 21-screw axes along [100] by C3\u2014H3\u22efO1 contacts (blue dotted lines). The remaining C\u2014H\u22efO as well as the C\u2014H\u22ef\u03c0 inter\u00adactions are displayed in Fig.\u00a03B\u22efO3 contacts (green dotted lines). These strands are linked by two different C\u2014H\u22ef\u03c0 contacts (Table\u00a01Cg is the centroid of this ring). Along [100] the strands are linked by C12\u2014H12\u22efiCgv contacts (orange dotted lines) while along [001] the links are established by C17\u2014H17B\u22efCgv inter\u00adactions (violet dotted lines) enclosing angles between the C\u2014H bond and the plane of the \u03c0-system of ca 39\u00b0 and 75\u00b0 respectively. As a result of these inter\u00adactions, a three-dimensional supra\u00admolecular structure is formed.The crystal packing of the title compound shows weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions Table\u00a01 and 3 \u25b8.et al., 1998trans-1,2-bis\u00ad(meth\u00adoxy\u00adcarbon\u00adyl)-6,6-di\u00admethyl\u00adspiro\u00ad(2.5)octane-4,8-dione -1,1-dichloro-6,6-dimethyl-2-[(1\u2032S*)-1\u2032-nitro\u00adeth\u00adyl]spiro\u00ad[2.5]octane-4,8-dione octane 1,4-dioxane solvate -5-benzyl-2,2,3-trimethyl-4-oxo-1-[(E)-3-phenyl\u00adallyl\u00adidene]-imidazolidin-1-ium hexa\u00adfluoro\u00adphosphate and phenyl\u00adiodo\u00adnium-4,4-di\u00admethyl\u00adcyclo\u00adhexane-2,6-dione in aceto\u00adnitrile (5\u2005ml). After 24\u2005h stirring at ambient temperature, water (10\u2005ml) was added. The aqueous phase was extracted with CH2Cl2 (15\u2005ml). The organic layers were combined, washed with brine, and dried over MgSO4. After evaporation of the solvent under vacuum, the crude product was purified by column chromatography (n-penta\u00adne/Et2O: 7/3 and 6/4) to give the title compound as colourless crystals (m.p. 397\u2013399\u2005K).A 10\u2005ml round-bottomed flask equipped with a magnetic stirring bar was charged with a solution of the (Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. The methyl groups were allowed to rotate along the C\u2014C bonds to best fit the experimental electron density. As a result of the absence of anomalous scatterers and high angle data, the Flack test results can be considered meaningless. The synthesis resulted in a racemic mixture, hence the structure was refined as an inversion twin.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901600164X/su5275sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901600164X/su5275Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901600164X/su5275Isup3.cmlSupporting information file. DOI: 1450224CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "If Mo\u2014Mo bonds are ignored, each Mo atom can be considered as six-coordinated in a distorted octa\u00adhedral geometry. The Mo\u2014Mo distance of 2.6880\u2005(5)\u2005\u00c5 for two the Mo atoms bridged by the acetate ligand is shorter than the other two Mo\u2014Mo distances [2.7490\u2005(5) and 2.7566\u2005(5)\u2005\u00c5]. One ethyl group is disordered between two conformations in a 0.65\u2005(3):0.35\u2005(3) ratio. In the crystal, weak C\u2014H\u22efO inter\u00adactions link the trinuclear mol\u00adecules related by translation in [100] into chains. The crystal packing exhibits short inter\u00admolecular S\u22efS contacts of 3.1886\u2005(13)\u2005\u00c5. In other words, in this crystal packing, a supramolecular structure is constructed by the C\u2014H\u22efO and S\u22efS interactions.In the title compound, [Mo DOI: 10.1107/S1600536814012501/cv5459Isup2.hklStructure factors: contains datablock(s) I. DOI: 1005735CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The least-squares planes through the two aromatic ring systems make an angle of 47.3\u2005(3)\u00b0 for (I) and 38.6\u2005(2)\u00b0 for (II). Both structures show disorder for the PtCl2 fragment, in the case of (I) even further extended towards the CH2\u2014CH=CH2 ligand. An intra\u00admolecular C\u2014H\u22efCl hydrogen bond occurs in (I). In the crystal packing of (I), which is dominated by N\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions, a partially occupied water mol\u00adecule is observed on a twofold rotation axis with a refined site occupancy of 0.10\u2005(1). A C\u2014H\u22ef\u03c0 inter\u00adaction is also present. In (II), inversion dimers form chains along the b-axis direction by N\u2014H\u22efO hydrogen bonds.In the title complexes, In both complexes, the Cl atoms are trans with respect to each other 2CH2=CH\u2014CH2 fragment is disordered over two positions [population parameters 0.679\u2005(8) and 0.321\u2005(8)], while in (II)2 fragment is disordered over two positions [population parameters 0.872\u2005(6) and 0.128\u2005(6)]. The angles between the best planes through the two aromatic rings are 47.3\u2005(3) and 38.6\u2005(2)\u00b0 for (I)A\u22efCl9 distance of 2.73\u2005\u00c5. In (I)A\u22efCl9.The complexes crystallize in different space groups, er Fig.\u00a01. The eugThe crystal packing of (I)AB\u22efO28i hydrogen bonds are further linked into chains in the b-axis direction by N2\u2014H2AA\u22ef hydrogen bonds .The crystal packing of (II)B\u22efCg1iii, H27B\u22efCg1iii = 2.72\u2005\u00c5; Cg1 is the centroid of the C20-C25 ring; symmetry code: (iii) \u2212x\u00a0+\u00a01, y, \u2212z\u00a0+\u00a0No \u03c0\u2013\u03c0 inter\u00adactions are observed in the packing of either structure. For (I)B) due to the disorder from the average Pt\u2014N distance of 2.09\u2005(5)\u2005\u00c5 for Pt\u2014NH2\u2014phenyl fragments present in the Cambridge Structural Database and 2.305\u2005(2)\u2005\u00c5 and agree well with the average Pt\u2014Cl distance of 2.32\u2005(3)\u2005\u00c5 for trans complexes present in the CSD. One Pt1B\u2014Cl distance [2.151\u2005(2)\u2005\u00c5] deviates significantly from this average.The Pt\u2014N distances in (I)2 and C=C shows 11 hits. As fourth ligand we notice an additional Cl atom or C atom (two hits) or O atom (one hit). In the complex [PtCl(methyl\u00adeugenol)(o-toluidine)] A search in the CSD for Pt complexes with Pt coordinated by Cl, NH3]:Synthesis of K[Pt(Alkeug)Cl3] were synthesized following the protocol of Da and coworkers Cl2(Alkeug)(C6H6NCl)]:Synthesis of trans-[PtClv/v) was added to a mixture of 1.0\u2005mmol [K[Pt(Alkeug)Cl3] and 10\u2005mL acetone/ethanol (1:1 v/v). After two\u2005h stirring, a white precipitate of KCl was separated out. The remaining solution was stirred for two\u2005h at room temperature to obtain a yellow precipitate, which was collected by filtration, washed with ethanol and diethyl ether and dried in vacuum. The obtained crystals are soluble in chloro\u00adform and acetone, slightly soluble in ethanol and insoluble in water. The yield was 70\u201380%. Single crystals suitable for X-ray investigation were obtained by slow evaporation from a chloro\u00adform/ethanol (1:2 v/v) solution at room temperature.A solution of 127.0\u2005mg (1.0\u2005mmol) p-chloro\u00adaniline in 10\u2005mL acetone/ethanol (1:1 2(Meteug)(C6H6NCl)] (I)Data for (II)Data for and refined with constraints for the bond lengths present in this fragment. Refinement of the population parameter of oxygen atom O34 converged to 0.10\u2005(1). Water H atoms were not located.In (I)2 fragment is disordered over two positions [population parameters 0.872\u2005(6) and 0.128\u2005(6)].In (II)Uiso(H) values assigned as 1.2Ueq of the parent atoms (1.5 times for methyl groups), with C\u2014H distances of 0.95 (aromatic and =CH2), 0.98 (CH3), 0.99 (CH2) and 1.00\u2005\u00c5 (CH), and N\u2014H distances of 0.91\u2005\u00c5 (NH2). Enhanced rigid bond restraints were used for the anisotropic temperature factors of the non-H atoms. In the final cycles of refinement, 7 and 15 outliers were omitted for (I)All H atoms were placed in idealized positions and refined in riding mode, with 10.1107/S2056989016008872/rz5189sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989016008872/rz5189Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016008872/rz5189IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1483067, 1483066CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The complexes each form tetrahedral C 2(C8H11BrO3)(CO)6], (1), and [Co2(C12H16Br2O4)(CO)6], (2), result from the replacement of two carbonyl ligands from dicobalt octa\u00adcarbonyl by the alkynes 4-hy\u00addroxy\u00adbut-2-ynyl 2-bromo-2-methyl\u00adpropano\u00adate and but-2-yne-1,4-diyl bis\u00ad(2-bromo-2-methyl\u00adpropano\u00adate), respectively. Both mol\u00adecules have classic tetra\u00adhedral C2Co2 cluster cores with the CoII atoms in a highly distorted octa\u00adhedral coordination geometry. The alkyne ligands both adopt a cis-bent conformation on coordination. In the crystal structure of (1), classical O\u2014H\u22efO and non-classical C\u2014H\u22efO contacts form inversion dimers. These combine with weak O\u22efO and Br\u22efO contacts to stack the mol\u00adecules into inter\u00adconnected columns along the b-axis direction. C\u2014H\u22efO and C\u2014H\u22efBr contacts stabilize the packing for (2), and a weak Br\u22efO contact is also observed. Inter\u00adconnected columns of mol\u00adecules again form along the b-axis direction.The title compounds, [Co The two carbonyl groups in (2) each lie on the same side of the mol\u00adecule, with the 2-bromo-2-methyl\u00adpropano\u00adate units arranged symmetrically with respect to the central Cet al., 1995A\u22efO1 and C8\u2014H8A\u22efO12 hydrogen bonds and Br1\u22efO1 contacts stack the mol\u00adecules into inter\u00adconnected columns along the b-axis direction , classical O1\u2014H1\u22efO3 hydrogen bonds Table\u00a03 are augmds Fig.\u00a03. These con Fig.\u00a04.B\u22efO2 and C8\u2013H8A\u22efO2 contacts \u2005\u00c5, symmetry operation x, \u22121\u00a0+\u00a0y, z], form b-axis direction , although in this mol\u00adecule no classical hydrogen bonds are possible. Bifurcated C3\u2014H3s Table\u00a04 produce gs Fig.\u00a05. The oth0] Fig.\u00a06. The neton Fig.\u00a07.et al., 2002The first structure, of dicobalt hexa\u00adcarbonyl di\u00adphenyl\u00adacetyl\u00adene, was reported using film data Sly, 1959. The cur2(CO)8 were allowed to react at room temperature for 1\u2005h in CH2Cl2 under nitro\u00adgen. The reaction mixtures were filtered through silica gel to remove any insol\u00aduble impurities and the filtrates taken to dryness in vacuo. The complexes were then purified by recrystallization from hexane at 273\u2005K. Yields were in the range 70\u201380%. Complexation was confirmed by the absence of a band at 1860\u2005cm\u22121 in the infrared spectrum, attributable to the \u03bc2 (bridging) carbonyl groups of the dicobalt octa\u00adcarbonyl starting material. In addition, a hypsochromic shift of approximately 30\u2005cm\u22121 of the remaining carbonyl stretching frequencies is seen, due to the decrease in electron density at the metal atoms upon coordination of these alkynes. Characteristic IR spectra were recorded for both products as follows: IR : (1): 3300 , \u03bd(C\u2261O) 2099, 2062, 2032, \u03bd(C=O) 1735; (2): \u03bd(C\u2261O) 2096, 2058, 2031, \u03bd(C=O) 1734.In typical preparations, 1:1 molar qu\u00adanti\u00adties of 4-hy\u00addroxy\u00adbut-2-ynyl 2-bromo-2-methyl\u00adpropano\u00adate for (1) or a 2:1 molar ratio of but-2-yne-1,4-diyl bis\u00ad(2-bromo-2-methyl\u00adpropano\u00adate) for (2) with Cod(C\u2014H) = 0.99\u2005\u00c5, Uiso = 1.2Ueq (C) for CH2, 0.98\u2005\u00c5, Uiso = 1.5Ueq (C) for CH3 atoms. In the final refinement, two reflections from the data for (2) with Fo << Fc were omitted from the refinement.All H atoms bound to carbon were refined using a riding model with 10.1107/S1600536814009659/hb0001sup1.cifCrystal structure: contains datablock(s) global, 1, 2. DOI: 10.1107/S1600536814009659/hb00011sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S1600536814009659/hb00012sup3.hklStructure factors: contains datablock(s) 2. DOI: 1004257, 1004258CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The benzyl\u00adidine and benzoate rings are inclined to one another by 24.58\u2005(8)\u00b0, and the conformation about the C=N bond is E.The title p-hy\u00addroxy Schiff base, C17H17NO4, was synthesized via the condensation reaction of benzocaine with vanillin. The benzyl\u00adidine and benzoate rings are inclined to one another by 24.58\u2005(8)\u00b0, and the conformation about the C=N bond is E. In the crystal, mol\u00adecules are linked by O\u2014H\u22efN hydrogen bonds, forming zigzag chains propagating along [010]. Adjacent chains are linked by C\u2014H\u22ef\u03c0 and weak offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.819\u2005(1)\u2005\u00c5], forming sheets parallel to (10-2).The title The m-meth\u00adoxy group (O1/C13/C16) is slightly out of the plane of the benzene ring (C11\u2013C14/C20/C21) to which it is attached by 5.37\u2005(18)\u00b0, while the mean plane of the ethyl\u00adacetate group (O3/O17/C1/C2/C4) is inclined to the benzene ring (C5\u2013C8/C18/C19) to which it is attached by 10.23\u2005(11)\u00b0. This non-linearity is consistent for Schiff bases.The title Schiff base, (I)Cg1\u22efCg1i = 3.819\u2005(1)\u2005\u00c5, inter\u00adplanar distance = 3.672\u2005(2)\u2005\u00c5, slippage = 1.05\u2005\u00c5, Cg1 is the centroid of ring C5\u2013C8/C18/C19; symmetry code: (i) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01],In the crystal, mol\u00adecules are linked by O\u2014H\u22efN hydrogen bonds, forming zigzag chains propagating along [010]; see Table\u00a01para rather than the ortho position, this Schiff base cannot form the intra\u00admolecular C=N\u22efO\u2014H hydrogen bond responsible for keto\u2013enol tautomerism. However, the close proximity of the C=N and O\u2014H groups gives rise to the possibility that external stimulation of the material by heat or light may lead to the zwitterionic form. The potential for compound (I)The crystal structure analysis of compound (I)et al., 2016et al., 2010et al., 2012In the Cambridge Structural Database Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016008999/su5304sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989016008999/su5304Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016008999/su5304sup3.pdfSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016008999/su5304Isup4.cmlSupporting information file. DOI: 1483394CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dihedral angles between the benzo\u00adthia\u00adzole ring systems and the phenol rings are 4.1\u2005(4) and 5.8\u2005(4)\u00b0, indicating an almost planar geometry. Weak intra- and inter\u00admolecular C\u2014H\u22efO hydrogen bonds are observed. In the crystal, weak \u03c0\u2013\u03c0 inter\u00adactions between aromatic and thia\u00adzole rings [centroid\u2013centroid distances = 3.626\u2005(3) and 3.873\u2005(3)\u2005\u00c5] link the mol\u00adecules into a two-dimensional supra\u00admolecular network along the bc plane.In the title complex, [Cu(C DOI: 10.1107/S2056989015015303/bq2400Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015015303/bq2400fig1.tif. DOI: Mol\u00adecular structure of the title complex, showing the atom-numbering scheme and 30% probability ellipsoids.Click here for additional data file.10.1107/S2056989015015303/bq2400fig2.tifvia . DOI: via C\u2014H\u22efO (black dashed lines) and \u03c0-\u03c0 (red) inter\u00adactions.Dimeric formation Click here for additional data file.10.1107/S2056989015015303/bq2400fig3.tif. DOI: Part of the crystal structure of the title complex, showing the 2-D network of mol\u00adecules linked by inter\u00admolecular C\u2014H\u22efO hydrogen bonds (black dashed lines) and \u03c0-\u03c0 inter\u00adactions (red).1419096CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The packing is stabilized 42H42N2P22+\u00b72Br\u2212\u00b72CH2Cl2, lies on a crystallographic twofold rotation axis. The 1,2-di\u00adamino\u00adcyclo\u00adhexane fragment has a chair conformation with two N atoms in a transoid conformation [N\u2014C\u2014C\u2014N = 163.4\u2005(2)\u00b0]. In the crystal, the cations are linked to the anions by N\u2014H\u22efBr and C\u2014H\u22efBr hydrogen bonds, forming a chain structure along the c axis. The di\u00adchloro\u00admethane mol\u00adecule takes part in the hydrogen-bond network through C\u2014H\u22ef\u03c0 and C\u2014H\u22efBr inter\u00adactions.The cation of the title solvated salt, C P,P,P-triaryl-P-amino\u00adphospho\u00adnium salts bearing a primary amino group are isolable inter\u00admediates in the Horner & Oediger \u00b0; symmetry code: (i) \u2212x\u00a0+\u00a01, y, \u2212z\u00a0+\u00a0The cation is situated on a crystallographic twofold rotation axis Fig.\u00a01. The 1,2trans-1,2-di\u00adamino\u00adcyclo\u00adhexane mol\u00adecule and the last is donated by the solvent di\u00adchloro\u00admethane mol\u00adecule in 100\u2005ml of the same solvent. After four\u2005h of stirring at room temperature and the formation of white precipitate, a mixture of half an equivalent of (\u00b1)-trans-1,2-di\u00adamino\u00adcyclo\u00adhexane and one equivalent of tri\u00adethyl\u00adamine in 10\u2005ml of CH2Cl2 was added dropwise under stirring at 273\u2005K. The suspension was left under continuous stirring for 12\u2005h at room temperature. Then the reactant was extracted twice with 25\u2005ml of distilled water, and the organic phase was dried over MgSO4. All volatiles were eliminated under vacuum, and the resulting light-yellow solid was stirred with Et2O overnight. After filtration, 6.0\u2005g of the title compound was obtained as a white powder . Single crystals suitable for X-ray diffraction were grown by slow evaporation of a di\u00adchloro\u00admethane solution at room temperature.Under an NUiso(H) = 1.2Ueq(N). Other H atoms were positioned geometrically (C\u2014H = 0.93 or 0.97\u2005\u00c5) and constrained using the riding-model approximation with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016004576/is5441sup1.cifCrystal structure: contains datablock(s) Global, I. DOI: 10.1107/S2056989016004576/is5441Isup2.hklStructure factors: contains datablock(s) I. DOI: 1469040CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The ruthenocenecarbo\u00adnitrile mol\u00adecule exhibits mirror symmetry in the solid state. 5-C5H4C\u00a0N)(\u03b75-C5H5)], exhibits point group symmetry m, with the mirror plane bis\u00adecting the mol\u00adecule through the C\u00a0N substituent. The RuII atom is slightly shifted from the \u03b75-C5H4 centroid towards the C\u00a0N substituent. In the crystal, mol\u00adecules are arranged in columns parallel to [100]. One-dimensional inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [3.363\u2005(3)\u2005\u00c5] between the C\u00a0N carbon atom and one carbon of the cyclo\u00adpenta\u00addienyl ring of the overlaying mol\u00adecule are present.The mol\u00adecular structure of ruthenocenecarbo\u00adnitrile, [Ru(\u03b7 A synthesis for ferrocenecarbo\u00adnitrile has already been described in 1957 \u2005\u00c5]. Both cyclo\u00adpenta\u00addienyl rings adopt an ideally eclipsed conformation and are virtually oriented parallel towards each other, which is expressed by the bond angle at the RuII between the two centroids (= D), with D(C5H4)\u2014Ru1\u2014D(C5H5) = 178.87\u2005(1)\u00b0. However, the RuII atom is slightly shifted from the centre of the C5 ring to the nitrile-bonded C2 atom, which can be explained best by the significantly different Ru\u2014C bond lengths sp (ethyn\u00adyl) as compared to (I)et al., 2008The ruthenocene backbone is hardly described in the literature. Reported derivatives contain et al., 2008et al., 2010et al., 1996et al., 2007et al., 2007Comparison of the C\u2014C [1.431\u2005(3)\u2005\u00c5] and the C\u00a0N distances [1.148\u2005(3)\u2005\u00c5] with the respective ferrocene carbo\u00adnitrile derivatives solution were added in a single portion, and stirring was continued for additional 20\u2005min. Solid particles were removed by filtration and the filtrate was extracted with ethyl acetate (3 \u00d7 50\u2005ml). The combined organic layers were dried over MgSO4. All volatiles were removed under reduced pressure and the crude product was purified by flash chromatography on aluminum oxide using di\u00adchloro\u00admethane as eluent. Greenish crystals of (I)\u22121): \u03bd = 2226 , 2854 (s), 2925 (s), 3082 . 1H NMR : \u03b4 4.69 , 4.70 , 4.70 . 13C{1H} NMR : \u03b4 = 55.3 (Ci-C5H4), 72.4 (C5H4), 72.9 (C5H5), 73.5 (C5H4), 119.4 (CN). HRMS : C11H9NRu: m/z = 256.9792 .Formyl\u00adruthenocene was prepared according to a published procedure (Mueller-Westerhoff isoU(H) = 1.2Ueq(C) and a C\u2014H distance of 0.93\u2005\u00c5. Crystal data, data collection and structure refinement details are summarized in Table\u00a02C-bonded H atoms were placed in calculated positions and constrained to ride on their parent atoms, with 10.1107/S205698901500540X/wm5119sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901500540X/wm5119Isup2.hklStructure factors: contains datablock(s) I. DOI: 1054219CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Zn\u22efZn separation is 11.3893\u2005(14)\u2005\u00c5 and the ligands wrap around the two ZnII atoms, forming a triple helix as defined by the Zn\u2014N\u2014N\u2014Zn torsion angles of 104.05\u2005(18), 99.06\u2005(19) and 101.40\u2005(19)\u00b0. The Zn\u2014N(pyrid\u00adyl) distances in the octahedral ZnN6 coordination sphere are in the range 2.128\u2005(5)\u20132.190\u2005(5)\u2005\u00c5 and the Zn\u2014N(imine) distances are in the range 2.157\u2005(5)\u20132.277\u2005(5)\u2005\u00c5.The asymmetric unit of the title compound, [Zn DOI: 10.1107/S205698901502455X/fk2093Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901502455X/fk2093fig1.tif. DOI: Mol\u00adecular structure of the title compound. Anisotropic displacement ellipsoids are drawn at the 50% probability level and H atoms are represented by circles of arbitrary size.1443648CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The [Re6Se8]2+ cluster core displays a non-crystallographic center of symmetry and is bonded through the ReIII atoms to five tri\u00adethyl\u00adphosphane ligands and one p-toluene\u00adsulfonate ligand. One p-toluene\u00adsulfonate counter-ion and two di\u00adchloro\u00admethane solvent mol\u00adecules are also present in the asymmetric unit. One of the ethyl chains of one triethylphos\u00adphane ligand and one of the CH2Cl2 solvent molecules are disordered over two sets of sites . The Re\u2014O(sulfon\u00adate) bond length of 2.123\u2005(5)\u2005\u00c5 is similar to other Re\u2014O bond lengths of hexa\u00adnuclear rhenium chalcogenide clusters containing other O-donor ligands such as dimethyl sulfoxide (DMSO), di\u00admethyl\u00adformamide (DMF) and hydroxide.The title compound, [Re Later, Eremenko et al. (1993iPr)3)2(CO)(NO)(OTs)2] (OTs\u2212 = p-toluene\u00adsulfonate anion) which represented the first structural report of a rhenium complex containing tosyl\u00adate ligands. In the synthesis of octa\u00adhedral rhenium chalcogenide cluster complexes, the substitution of either halide or nitrile ligands has proven an effective means for generating a variety of new cluster complexes cluster complexes (C6H15P)5](C7H7O3S)\u00b72CH2Cl2 = 0.036wR(F2) = 0.097S = 1.0216405 reflections693 parametersH-atom parameters constrainedmax = 2.67 e \u00c5\u22123\u0394\u03c1min = \u22121.26 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: DIRDIF99 I, isu0804. DOI: 10.1107/S2056989015014334/pj2021Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015014334/pj2021fig1.tif6 8 3 5 3 6 4 + . DOI: 6Se8(PEt3)5(O3SC6H4Me)]+ ion showing the atom labelling scheme. Non-hydrogen atoms are represented by Gaussian ellipsoids at the 30% probability level. Hydrogen atoms omitted for clarity.Perspective view of the [Re1010097CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Each water mol\u00adecule is disordered over three sets of sites, with a refined occupancy ratio of 0.310\u2005(9):0.275\u2005(9):0.415\u2005(9) for one mol\u00adecule and 0.335\u2005(9):0.288\u2005(9):0.377\u2005(9) for the other mol\u00adecule. The MnII atoms exhibit a distorted octa\u00adhedral geometry, while the WV atoms adopt a distorted square-anti\u00adprismatic geometry. The MnII and WV atoms are linked alternatively through cyanide groups, forming a tetra\u00adnuclear 12-atom rhombic metallacycle. Adjacent metallacycles are further connected by \u03bc2-bridging cyanide anions, generating a 3,2-chain structure running parallel to [101]. Inter\u00adchain \u03c0\u2013\u03c0 inter\u00adactions are observed [centroid\u2013centroid distances = 3.763\u2005(3) and 3.620\u2005(2)\u2005\u00c5].The asymmetric unit of the title compound, {[Mn DOI: 10.1107/S1600536814007235/rz5113Isup2.hklStructure factors: contains datablock(s) I. DOI: 994877CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N2-amino group of the guanine residue exists in the cyclic amino\u00adglycoside form.Crystallographic analysis of a nucleoside analog of the 2\u2032-de\u00adoxy\u00adguanosine/abasic site cross-link is presented. This structure corroborates an earlier two-dimensional NMR analysis, concluding that the 2-de\u00adoxy\u00adribose unit attached at the exocyclic R,4S,5R)-4-hy\u00addroxy-5-(hy\u00addroxy\u00admeth\u00adyl)tetra\u00adhydro\u00adfuran-2-yl]-2-{[-4-meth\u00adoxy-5-(meth\u00adoxy\u00admeth\u00adyl)tetra\u00adhydro\u00adfuran-2-yl]amino}-1H-purin-6(9H)-one, C17H25N5O7, crystallizes with two independent mol\u00adecules (A and B) in the asymmetric unit. In the crystal, the guanosine moieties of mol\u00adecules A and B are linked by N\u2014H\u22efN and O\u2014H\u22efN hydrogen-bonding inter\u00adactions, forming ribbons which are stacked to form columns along [100]. These columns are then linked by O\u2014H\u22efO hydrogen bonds between the ribose moieties and numerous C\u2014H\u22efO inter\u00adactions to complete the three-dimensional structure.The title compound, 9-[(2 The disposition of these furan\u00adose rings relative to the purine rings can be described by the torsion angle C2\u2014N4\u2014C6\u2014O2, which is 70.9\u2005(3)\u00b0 in mol\u00adecule A and 61.7\u2005(3)\u00b0 in mol\u00adecule B. The furan\u00adose ring attached to the N5 position in mol\u00adecule A is again a half-chair, with the maximum deviation from planarity between C11A and C12A , while this furan\u00adose ring in mol\u00adecule B is an envelope with C11B at the flap . The disposition of these furan\u00adose rings relative to the purine rings can be described by the angle C1\u2014N5\u2014C11\u2014O5, which is \u221287.4\u2005(2)\u00b0 in mol\u00adecule A and \u221293.7\u2005(2)\u00b0 in mol\u00adecule B.The two independent mol\u00adecules were dissolved in 0.8\u2005ml of a 3:1 mixture of DMSO and 25\u2005mM sodium phosphate buffer (pH 7.0) in a round-bottom flask. The flask was heated to 333\u2005K and the mixture stirred for 22\u2005h. The solvent removed in vacuo and the product purified by column chromatography on silica gel eluted with 0\u201315% methanol in di\u00adchloro\u00admethane to yield 36\u2005mg (12% yield) of the title compound as a colorless oil. The precursor 3,5-bis-O-methyl-2-de\u00adoxy-d-ribo\u00adfuran\u00adose was synthesized according to previously reported procedures and 3,5-bis-Uiso(H) = 1.2Ueq(C). Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901600517X/hb7568sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901600517X/hb7568Isup2.hklStructure factors: contains datablock(s) I. DOI: 1448235CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A stable oxopiperidinium trication salt was synthesized. In the crystal, N\u2014H\u22efCl, C\u2014H\u22efCl and C\u2014H\u22efO hydrogen bonds link cations and anions into a three-dimensional network. 28H40N3O3+\u00b73Cl\u2212\u00b72H2O, the central heterocyclic ring adopts a sofa conformation, with the exocyclic N\u2014C bond in an equatorial orientation. The dihedral angles between the planar part of this heterocyclic ring and the two almost flat side-chain fragments, which include the aromatic ring and bridging atoms, are 28.8\u2005(1) and 41.1\u2005(1)\u00b0. Both di\u00adethyl\u00adaza\u00adniumyl substituents have a tetra\u00adhedral geometry, while the dihedral angles between the above-mentioned flat part of the aryl fragments and the imaginary planes drawn through atoms C\u2014N\u2014C of the di\u00adethyl\u00adaza\u00adniumyl substituents are 86.3\u2005(2) and 80.4\u2005(1)\u00b0, respectively. In the crystal, N\u2014H\u22efCl hydrogen bonds link the cations and anions into [100] chains. The chains are cross-linked by numerous C\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions, generating a three-dimensional network. One of the chloride ions is disordered over two adjacent positions in a 0.895\u2005(4):0.105\u2005(4) ratio.In the trication of the title hydrated mol\u00adecular salt, C The dihedral angles between the flat part of the heterocycle and the two almost planar fragments that include the phenyl-ring and the bridging atoms are 28.7\u2005(1) and 41.1\u2005(1)\u00b0 for (C7\u2013C13) and (C18\u2013C24), respectively. Such non-planarity might partly be caused by the presence of short intra\u00admolecular contacts H2AB\u22efH24A and H6AB\u22efH13A with distances 2.18 and 2.14\u2005\u00c5, respectively, which are shorter than the doubled van der Waals radius of the H atom -3,5-bis\u00ad[4-(di\u00adethyl\u00adamino)\u00adbenzyl\u00adidene[\u22121-methyl-4-piperidone was obtained according to a literature procedure = k \u00d7 Ueq(C), where k = 1.2 for CH and CH2 and 1.5 for CH3 H atoms. All N-bound H atoms were located using difference Fourier maps, but in the final refinement their distances were constrained at 0.90\u2005\u00c5 (DFIX). H atoms of the two water mol\u00adecules were not localized properly, since they appeared to be disordered over several positions. These H atoms were therefore removed from the refinement, but they were still included in the resulting chemical formula. Atom Cl3 is disordered over two positions in a 0.895\u2005(4):0.105\u2005(4) ratio.Crystal data, data collection, and structure refinement details are summarized in Table\u00a0210.1107/S2056989015020952/hb7503sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015020952/hb7503Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015020952/hb7503Isup3.cmlSupporting information file. DOI: 1435161CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title aryl\u00adsulfonyl glycinyl hydrazone Schiff base compound crystallizes as a monohydrate. In the crystal, a series of O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds leads to the formation of corrugated sheets lying parallel to (100). 16H16ClN3O3S\u00b7H2O, is L-shaped being bent at the S atom; the S\u2014N\u2014C\u2014C torsion angle is 132.0\u2005(3)\u00b0. The central part of the mol\u00adecule, C\u2014C\u2014N\u2014N=C, is almost linear, with the C\u2014C\u2014N\u2014N and C\u2014N\u2014N=C torsion angles being \u2212174.1\u2005(2) and 176.0\u2005(2)\u00b0, respectively. The dihedral angle between the p-toluene\u00adsulfonyl ring and the S\u2014N\u2014C\u2014C(=O) segment is 67.5\u2005(4)\u00b0, while that between the two aromatic rings is 52.17\u2005(11)\u00b0. In the crystal, the water H atom is involved in O\u2014H\u22efO hydrogen bonds with a sulfonamide O atom and the carbonyl O atom. The water O atom is itself hydrogen bonded to both NH hydrogen atoms. These four hydrogen bonds lead to the formation of corrugated sheets lying parallel to (100). There are also weak C\u2014H\u22efO contacts present within the sheets.The mol\u00adecule of the title compound, C N-Acyl\u00adhydrazones containing a glycine residue have been investigated extensively in recent years for their biological and medical activities -amides \u2005\u00c5 indicates that the mol\u00adecule exists in the keto form in the solid state. The C9\u2014N3 bond length of 1.266\u2005(3)\u2005\u00c5 confirms its significant double-bond character. The N2\u2014N3 and C8\u2014N2 bond distances are 1.384\u2005(3) and 1.337\u2005(3)\u2005\u00c5, respectively, which indicate significant delocalization of the \u03c0-electron density over the hydrazone portion of the mol\u00adecule. The mol\u00adecule is bent at the S-atom with a S1\u2014N1\u2014C7\u2014C8 torsion angle of 132.0\u2005(2)\u00b0. The other central part of the mol\u00adecule is almost linear with the C7\u2014C8\u2014N2\u2014N3, C8\u2014N2\u2014N3\u2014C9 and N2\u2014N3\u2014C9\u2014C10 torsion angles being \u2212174.1\u2005(2), 176.0\u2005(2) and \u2212176.7\u2005(2)\u00b0, respectively. The orientation of the sulfonamide group with respect to the attached p-toluene\u00adsulfonyl ring (C1\u2013C6) is given by torsion angles C2\u2014C1\u2014S1\u2014N1 = \u221299.8\u2005(2)\u00b0 and C6\u2014C1\u2014S1\u2014N1 = 76.6\u2005(2)\u00b0, while that of the hydrazone group with the attached benzene ring (C10-C15) is given by torsion angles C11\u2014C10\u2014C9\u2014N3 = 9.9\u2005(4)\u00b0 and C15\u2014C10\u2014C9\u2014N3 = \u2212172.1\u2005(2)\u00b0. The dihedral angles between the mean plane of the central segment and the benzene rings (C1\u2013C6 and C10\u2013C15) are 65.22\u2005(15) and 13.06\u2005(14)\u00b0, respectively. The two benzene rings are inclined to one another by 52.16\u2005(14)\u00b0.The mol\u00adecular structure of the title compound is illustrated in Fig.\u00a01In the crystal, the water O-atom, O4, shows bifurcated hydrogen bonding with the amino-H atom of the hydrazide segment (N2) and the sulfonamide-H atom (N1); see Table\u00a01viz. \u2013NH\u2013CH2\u2013C(=O)\u2013NH\u2013N=CH\u2013, yielded only one hit, namely N-(2-hy\u00addroxy-1-naphthyl\u00admethyl\u00adene)-N\u2032-(N-phenyl\u00adglyc\u00adyl)hydrazine was added to glycine (0.02\u2005mol) dissolved in an aqueous solution of potassium carbonate . The reaction mixture was stirred at 373\u2005K for 6\u2005h, then left overnight at room temperature, filtered and then treated with dilute hydro\u00adchloric acid. The solid N-(4-methyl\u00adbenzene\u00adsulfon\u00adyl)glycine (L1) obtained was crystallized from aqueous ethanol.The title compound was synthesized in a number of steps. Firstly L1 (0.02\u2005mol) dissolved in ethanol (30\u2005ml) and the mixture was refluxed. The reaction was monitored by TLC at regular inter\u00advals. After completion of the reaction, the reaction mixture was concentrated to remove excess ethanol. The product, N-(4-methyl\u00adbenzene\u00adsulfon\u00adyl)glycine ethyl ester (L2) obtained was poured into water, neutralized with sodium bicarbonate and recrystallized from acetone.Sulfuric acid (0.5\u2005ml) was added to L2 (0.01\u2005mol) was then added in small portions to a stirred solution of 99% hydrazine hydrate (10\u2005ml) in 30\u2005ml ethanol and the mixture was refluxed for 6\u2005h. After cooling to room temperature, the resulting precipitate was filtered, washed with cold water and dried to obtain N-(4-methyl\u00adbenzene\u00adsulfon\u00adyl)glycinyl hydrazide (L3).The pure L3 (0.01\u2005mol) and 3-chloro\u00adbenzaldehyde (0.01\u2005mol) in anhydrous methanol (30\u2005ml) and two drops of glacial acetic acid was refluxed for 8\u2005h. After cooling, the precipitate was collected by vacuum filtration, washed with cold methanol and dried. It was recrystallized to constant melting point from methanol (457\u2013458\u2005K). The purity of the title compound was checked and characterized by its IR spectrum. The characteristic absorptions observed are 3253.9, 1680.0, 1597.1, 1334.7 and 1161.2\u2005cm\u22121 for the stretching bands of N\u2014H, C\u2014O, C\u2014N, S\u2014O asymmetric and S\u2014O symmetric, respectively.A mixture of Prism-like colourless single crystals of the title compound were grown from a DMF solution by slow evaporation of the solvent.Uiso(H) = 1.5Ueq(O). The Ueq of atoms O1 and O2 were restrained to approximate isotropic behaviour. The NH H atoms were also located in a difference Fourier map and refined with Uiso(H) = 1.2Ueq(N). The C-bound H atoms were positioned with idealized geometry and refined using a riding model: C\u2014H = 0.93\u20130.97\u2005\u00c5 with Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015008506/su5128sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015008506/su5128Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015008506/su5128Isup3.cmlSupporting information file. DOI: 1062518CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound exists in the keto\u2013amine tautomeric form. In the crystal, supra\u00admolecular layers are formed by O\u2014H\u22efO hydrogen bonding. 12H16BrNO5 iminium\u00adyl}meth\u00adyl)-6-meth\u00adoxy\u00adbenzen-1-olate], C12H16BrNO5, is found in the keto\u2013amine tautomeric form, with an intra\u00admolecular iminium-N\u2014H\u22efO(phenolate) hydrogen bond and an E conformation about the C=N bond. Both gauche (two) and anti relationships are found for the methyl\u00adhydroxy groups. In the crystal, a supra\u00admolecular layer in the bc plane is formed via hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) and charge-assisted hy\u00addroxy-O\u2014H\u22efO(phenolate) hydrogen-bonding inter\u00adactions; various C\u2014H\u22efO inter\u00adactions provide additional cohesion to the layers, which stack along the a axis with no directional inter\u00adactions between them. A Hirshfeld surface analysis confirms the lack of specific inter\u00adactions in the inter-layer region.In the solid state, the title compound, C Thus, various species have been studied for their anticancer potential, Database survey. The conformation about the iminium bond [1.295\u2005(4)\u2005\u00c5] is E and this residue is almost coplanar with the benzene ring, forming a C2\u2014C1\u2014C7\u2014N1 torsion angle of 1.9\u2005(4)\u00b0. This arrangement allows for the formation of a tight charge-assisted iminium-N\u2014H\u22efO(phenolate) hydrogen bond (Table\u00a01gauche relationships about the C8\u2014C9 and C8\u2014C11 bonds , and an anti relationship about the C8\u2014C10 bond . The meth\u00adoxy group is almost coplanar with the ring it is connected to, as seen in the value of the C12\u2014O5\u2014C3\u2014C2 torsion angle of 177.7\u2005(2)\u00b0.The mol\u00adecular structure of (I)d Table\u00a01. The cona). Within this framework are a number of C\u2014H\u22efO inter\u00adactions, i.e. imine-C7\u2014H\u22efO(phenolate), methyl\u00adene-C11\u2014H\u22efO(phenolate) and methyl\u00adene-C9\u2014H\u22efO(hy\u00addroxy) . In accord with the distance criteria in PLATON As anti\u00adcipated from the chemical composition of (I)y) Fig.\u00a02b. In acdnorm contact distance within the range of \u22120.67 to 1.31\u2005\u00c5 through calculation of the inter\u00adnal (di) and external (de) Hirshfeld surface distances to the nearest nucleus over the range of \u22120.122 to 0.189 au. All Hirshfeld surface and fingerprints plots were generated using Crystal Explorer a), there are several red spots observed on the Hirshfeld surface of (I)deversus di , with the sum of contact distances being approximately 1.74\u2005\u00c5, signifying a strong inter\u00admolecular inter\u00adaction. Such strong inter\u00adactions constitute the second major contribution to the Hirshfeld surface, i.e. 25.4%, between the most prominent H\u22efH (38.2%) and other major contacts, like C\u22efH/H\u22efC (15.2%) and Br\u22efH/H\u22efBr (14.3%) and 3(d), C\u22efH and Br\u22efH contacts are at distances greater than their respective van der Waals radii. Fig.\u00a05The Hirshfeld surface map provides a visual summary of any close contacts (shown as red) in contrast to relatively long contacts (shown as white and blue). As displayed in Fig.\u00a03i Fig.\u00a03b, with %) Fig.\u00a04. Their ca), shows that the electronegative sites are predominantly converged on O atoms and that, upon crystallization, the electronegative and electropositive sites are connected . It is noteworthy that despite bromine being an electrophilic element, it did not form a significant non-covalent inter\u00adaction with neighbouring mol\u00adecules in the inter-layer region where these atoms are directed. The closest contact in this region occurs with methyl-C\u22efH12Ci, at 3.12\u2005\u00c5, i.e beyond the sum of the respective van der Waals radii , exists in the keto\u2013amine tautomeric form and has been the subject of several investigations \u00b0 in (I)There are several closely related structures to (I)12H16BrNO5: C 44.48, H 3.70, N 1.99%; found: C 44.81, H 3.42, N 1.64%. IR (cm\u22121): 3330 (b) \u03bd, 1640 (s) \u03bd(C=N), 1528 (m) \u03bd(\u2014O\u2014C=C\u2014), 1066 (m) \u03bd(C\u2014O\u2014C). 1H NMR : \u03b4 8.35 , 7.01\u20137.10 , 6.83\u20136.89 , 5.06 , 3.95 , 3.37\u20133.75 .A solution of tris\u00ad(hy\u00addroxy\u00admeth\u00adyl)amino\u00admethane was added to an ethano\u00adlic solution of 5-bromo-3-meth\u00adoxy-2-hy\u00addroxy\u00adbenzaldehyde and refluxed for 2\u2005h. The solution was allowed to stand at room temperature, during which an orange solid formed. This was recrystallized by slow evaporation of its ethanol solution. Yield: 2.67 (80%). Yellow crystals. M.p. 465\u2013466\u2005K. Analysis calculated for CUiso(H) set at 1.2\u20131.5Ueq(C). The O- and N-bound H atoms were located from difference Fourier maps and refined with distance restraints O\u2014H = 0.82\u00b10.01\u2005\u00c5 and N\u2014H = 0.86\u00b10.01\u2005\u00c5, and with Uiso(H) set at 1.5Ueq(O) and Uiso(H) set at 1.2Ueq(N), respectively. Owing to poor agreement, several reflections, i.e. (\u22129 7 7), (\u221212 4 6), (\u221210 5 6) and (\u22123 3 2), were omitted from the final cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016012159/hb7605sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016012159/hb7605Isup2.hklStructure factors: contains datablock(s) I. DOI: 1496206CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The complex exhibits twofold rotation symmetry, with the rotation axis bis\u00adecting the zinc cation. The structure is stabilized by weak inter\u00admolecular C\u2014H\u22efCl inter\u00adactions [C\u22efCl = 3.411\u2005(2) and 3.675\u2005(2)\u2005\u00c5], connecting neighbouring mol\u00adecules into layers perpendicular to the c axis.The structure of the title complex, [ZnCl DOI: 10.1107/S2056989015019143/zl2646Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015019143/zl2646fig1.tif11 9 3 2 2 x y -z . DOI: 11H9N3)2Cl2] with thermal ellipsoids plotted at the 30% probability level. Non-labelled atoms are created by the twofold symmetry axis .Mol\u00adecular structure of [Zn2Cl2] plotted down c, formed through weak C\u2013H\u22efCl inter\u00adactions.Two\u2013dimensional inter\u00adaction sheet of [Zn2Cl2] plotted down b axis, formed through weak C\u2013H\u22efCl inter\u00adactions.Two\u2013dimensional inter\u00adaction sheet of [Zn. H atoms are omitted for clarity.The arrangement of two-dimensional layers plotted down the 1430587CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Fe\u2014Fe), [Fe2(C8H8O)(CO)6], the diiron hexa\u00adcarbonyl moiety has a sawhorse arrangement, with the OC\u2014Fe\u2014Fe\u2014CO fragment forming the horizontal bar of the horse, and the other four carbonyl groups the legs. The Fe\u2014Fe distance is 2.795\u2005(2)\u2005\u00c5. Each Fe atom is also bonded to three C atoms of the cyclo\u00adocta\u00adtrienone ring. One Fe atom forms a \u03c3-bond with one ring C atom, with Fe\u2014C = 2.109\u2005(2)\u2005\u00c5, and also a metal\u2013olefin \u03c0-bond with two C atoms on the other side of the ring, with Fe\u2014C distances of 2.238\u2005(2) and 2.236\u2005(3)\u2005\u00c5. The second Fe atom forms a \u03b73-allyl bond with three other ring atoms, with Fe\u2014C bond lengths of 2.158\u2005(2), 2.062\u2005(2), and 2.123\u2005(3)\u2005\u00c5. Counting the \u03c0- and \u03c0-allyl inter\u00adactions as one bond, the coordinations of the Fe atoms can, respectively, be approximated as octa\u00adhedral and trigonal bipyramidal.In the title compound, bis\u00ad(tri\u00adcarbonyl\u00adiron)( I. DOI: 10.1107/S1600536814012690/pk2527Isup2.hklStructure factors: contains datablock(s) I. DOI: 1006076CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Ni\u2014N bond lengths to the pyridine rings are 2.1189\u2005(17) and 2.1241\u2005(17)\u2005\u00c5, whereas those to the thiocyanate anions are 2.0299\u2005(18) and 2.0359\u2005\u00c5. The Ni\u2014S bond lengths are 2.5357\u2005(6) and 2.5568\u2005(6)\u2005\u00c5. The Ni2+ cations are linked by N:S-bridging thio\u00adcyanate ligands into chains extending along [010]. The Ni\u22efNi distance within the chains is 5.5820\u2005(5)\u2005\u00c5. The asymmetric unit contains two Ni2+ cations of which one is located on a centre of inversion, whereas the second is located on a general position.In the title compound, [Ni(NCS) DOI: 10.1107/S1600536814009611/fk2081Isup2.hklStructure factors: contains datablock(s) I. DOI: 1000026CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "However, their intrinsic differences remain unclear. In this study, we analyzed gene expression profiles of the two subsets using Illumina HiSeq 2000 Sequencer. We identified 1995 transcripts related to the activation of V\u03b31+ \u03b3\u03b4 T cells, and 2158 transcripts related to the activation of V\u03b34+ \u03b3\u03b4 T cells. We identified 24 transcripts differentially expressed between the two subsets in resting condition, and 20 after PMA/Ionomycin treatment. We found that both cell types maintained phenotypes producing IFN-\u03b3, TNF-\u03b1, TGF-\u03b2 and IL-10. However, V\u03b31+ \u03b3\u03b4 T cells produced more Th2 type cytokines, such as IL-4 and IL-5, while V\u03b34+ \u03b3\u03b4 T cells preferentially produced IL-17. Our study provides a comprehensive gene expression profile of mouse peripheral V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells that describes the inherent differences between them.Peripheral \u03b3\u03b4 T cells in mice are classified into two major subpopulations, V\u03b31 Although considerable progress has been made in characterizing their biological significance, much remains unknown. \u03b3\u03b4 T cells arise earlier than \u03b1\u03b2 T cells during thymic ontogeny, predominately at the early stage of fetal development + and V\u03b34+ \u03b3\u03b4 T cells, depending on their TCR expression + and V\u03b34+ \u03b3\u03b4 T cells perform distinct functions in many disease models. For example, V\u03b31+ \u03b3\u03b4 T cells produce IL-4 and IFN-\u03b3 in the liver + \u03b3\u03b4 T cells produce IFN-\u03b3 or IL-17 depending on the studied models + and V\u03b34+ \u03b3\u03b4 T cells function as oppositional pairs in diseases including coxsackievirus B3 infection + and V\u03b34+ \u03b3\u03b4 T cells remains unresolved, partly due to a lack of comprehensive analysis and comparison of gene expression. Although, gene-expression profiles of emergent \u03b3\u03b4TCR+ thymocytes have been reported + and V\u03b34+ \u03b3\u03b4 T cells functional differences has not been reported. This is likely due to the limited number of cells that can be obtained from healthy mice.\u03b3\u03b4 T cells exert diverse functions, however, individual subsets within the population appear to be biased toward specialized functions + and V\u03b34+ \u03b3\u03b4 T cells simultaneously from the same pool of mouse splenocytes. We comprehensively analyzed gene expression profiles using Illumina\u2019s sequencing technology. We identified 1995 transcripts related to the activation of V\u03b31+ \u03b3\u03b4 T cells, and 2158 transcripts were related to the activation of V\u03b34+ \u03b3\u03b4 T cells. Interestingly, only 24 transcripts were differentially expressed between two subsets in resting condition, and 20 transcripts after PMA/Ionomycin-induced activation. Both cells produced high levels of IFN-\u03b3, TNF-\u03b1, TGF-\u03b2 and IL-10. However, V\u03b31+ \u03b3\u03b4 T cells produced more Th2 type cytokines, while V\u03b34+ \u03b3\u03b4 T cells tended to produce more IL-17. These findings describe the inherent differences between V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells.In this study, we expanded V\u03b31Male C57BL/6J mice aged 6\u20138 weeks were purchased from the National Institute for Food and Drug Control. All mice were maintained under specific pathogen-free conditions in the Experimental Animal Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences. All animal experiments were approved by and performed in accordance with the guidelines of the international Agency for Research on Cancer\u2019s Animal Care and Use Committee and IBMS/PUMC\u2019s Animal Care and Use Committee.+ and V\u03b34+ \u03b3\u03b4 T cells were expanded from splenocytes as described previously 7 cells were eluted and added to the Ab-coated wells (4\u00d7106 cells/well) and cultured in RPMI 1640 medium (Gibco BRL) supplemented with 10% fetal calf serum and IL-2 (200 IU/ml). After 8 days of expansion, the proportion of \u03b3\u03b4 T cells reached approximately 80% as determined by Flow Cytometry.V\u03b317 V\u03b31+ and 1.2\u00d7107 V\u03b34+ \u03b3\u03b4 T cells were sorted by Flow Cytometric Cell Sorting (FACS) with PE conjugated anti-mouse TCR V\u03b31.1/Cr4 antibody and APC conjugated anti-mouse TCR V\u03b32 antibody . The purity of sorted cells was more than 99%. 5\u00d7106 cells per well were seeded into 6-well culture plates at a concentration of 1\u00d7106/ml and rested overnight at 37\u00b0C in 5% CO2 in RPMI with 10% FCS. Cells were stimulated for 4 h with PBS or 20 ng/ml of PMA (Sigma) and 0.5\u00b5g/ml of Ionomycin (Sigma). Cells were washed with PBS and pelleted by centrifugation. Total RNA from each sample was extracted by Trizol reagent (Invitrogen) according to the manufacturer\u2019s instructions. The quality of total RNA from each sample was confirmed and comparable, based on results of Agilent Technologies 2100 Bioanalyzer.1.0\u00d710We prepared the Illumina libraries according to the manufacturer\u2019s instructions. Briefly, mRNAs were extracted from total RNA by mRNA enrichment kit followed by fragmentation of mRNA into 250\u2013350 bp sizes. The first strand cDNAs were synthesized using reverse transcriptase and random primers. Second strand cDNAs were synthesized using DNA Polymerase I followed by the addition of a single A base at the ends for the ligation to the adapters. After purification, the final cDNA library was created by PCR. Finally, 400\u2013500 bp products were used for cluster generation, 36 bp single-end sequencing was performed using Illumina HiSeq 2000 Sequencer according to the manufacturer\u2019s instructions (Beijing Berry Genomics Co. Ltd. China). The RNA-Seq raw data files have been deposited in NCBI\u2019s Sequence Read Archive (SRA) and are accessible through SRA Series accession number SRP042029.We performed base calling using CASAVA 1.7 software (Illumina). Low quality and polluted adapter reads were filtered; clean reads were stored on fastq files. The sequence reads were aligned to the mouse genome (mm9), and gene expression was calculated by RPKM value. Differentially expressed transcripts were identified using General Chi-square test analysis. Q values were obtained by the \u201cBH\u201d method + and V\u03b34+ \u03b3\u03b4 T cells were selected for verification from biological replicates with real-time quantitative PCR. RNA was extracted as described above. 500 ng of total RNA was reverse transcribed using PrimeScript RT reagent Kit with gDNA Eraser (Takara Bio). Gene-specific primers are listed in as described previously + and V\u03b34+ \u03b3\u03b4 T cells comprised approximately 35% and 25% respectively . \u03b3\u03b4 T cells were not screwed to one preferential subset after in vitro expansion, suggesting that in vitro expanded \u03b3\u03b4 T cells with anti-mouse TCR\u03b3\u03b4 antibodies plus IL-2 were still representative of in vivo subsets of \u03b3\u03b4 T cells. Expanded V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells were then sorted by FACS with PE-conjugated anti-mouse TCR V\u03b31.1/Cr4 antibody and APC conjugated anti-mouse TCR V\u03b32 antibody. We found the purities of sorted V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells were more than 99% or 20 ng/ml of PMA+0.5 \u00b5g/ml of Ionomycin (activated) before mRNA extraction and fragmentation. After cDNA synthesis, adapter ligation and PCR amplification, four cDNA libraries were constructed for the resting and activated \u03b3\u03b4 T cell subsets. 400\u2013500 bp-sized products were used for cluster generation and 36 bp single-end sequencing was performed by using Illumina HiSeq 2000 Sequencer. Approximately 28 million clean reads were obtained from each sample. More than 88% of reads were mapped to the mouse genome using the default setting in TopHat, suggesting high quality of RNA-seq . Cufflin+ and V\u03b34+ \u03b3\u03b4 T cells share similar transcript profiles in both the resting and activated subsets. We identified 24 transcripts with differential expression between the resting V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells (PMA/Ionomycin treatment induces a robust non-TCR mediated response in \u03b3\u03b4 T cells (q<0.05) . 2,158 t(q<0.05) . For a g(q<0.05) .+ \u03b3\u03b4 T cells were enriched for 32 KEGG pathways (p<0.05) (+ \u03b3\u03b4 T cells were enriched for 29 KEGG pathways (p<0.05) . 2,158 t(p<0.05) . Our com+ and V\u03b34+ \u03b3\u03b4 T cells , Itgb2 (Cd18), Itgal (Cd11a), Itgae (Cd103) and Itgb1 (Cd29) (Resting V\u03b31 subsets . Interes1 (Cd29) . However+ and V\u03b34+ \u03b3\u03b4 T cells, upregulating T cell activation markers CD25, CD69 and CD44 along with several cytokines. Therefore, we analyzed the expression of these representative markers in activated V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells. As expected, PMA/Ionomycin treatment induced expression of XCL1, CCL3, CCL4, CCL1, IFN-\u03b3, Lta, Csf2, TNF-\u03b1, IL-2, Gzmb and Gzmc technology. They concluded inherent gene expression differences in subsets defined their distinct functional responses Phylogenetic analysis suggests \u03b3\u03b4 T cells are precursors to modern B and \u03b1\u03b2 T cells + and V\u03b34+ \u03b3\u03b4 T cells are major subpopulations of peripheral \u03b3\u03b4 T cells in mice. Although global gene expression profiles of all emergent \u03b3\u03b4 thymocyte subsets have been reported by the Immunological Genome (ImmGen) Project and much knowledge has been obtained about the early divergence of gene expression programs between different \u03b3\u03b4 thymocyte subsets + and V\u03b34+ \u03b3\u03b4 T cells isn\u2019t available. A major hurdle has been the limited number of cells that can be obtained from healthy mice.V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells simultaneously from a single pool of mouse splenocytes. Our results proved that in vitro TCR-induced expansion for a week did not significantly change the proportion of V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells. We provide a comprehensive gene expression profile of mouse peripheral V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells in the resting and activated state. Although V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells share similar transcript profiles, we identified subset specific genes defining characteristics of each subset.In this study, we resolved the limited cell count issue by establishing a primary culture method expanding V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells, and 20 transcripts differentially expressed after PMA/Ionomycin treatment. Consistent with \u03b3\u03b4 thymocytes, expression levels of Rorc, Sox13 and Scart2 were higher in V\u03b34+ \u03b3\u03b4 T cells compared with V\u03b31+ \u03b3\u03b4 T cells Rorc expression is reported in \u03b3\u03b4 T cells, Th22 cells, NKT cells, CD4+ CD8+ thymocytes, and others that do not belong to the T or B cell lineage Rorc is recognized as a lineage-specific transcription factor of Th17 and is also required for IL-17 production Sox13 serves a general role in the differentiation of \u03b3\u03b4 T cells Sox13 was indispensable for the maturation of V\u03b34+ Th17 cells Scart2 is a marker of \u03b3\u03b4 T cells prepared to secrete IL-17A + \u03b3\u03b4 T cells compared with V\u03b31+ \u03b3\u03b4 T cells produce significantly more IL-17A and IL-17F after PMA/Ionomycin treatment are also consistent with findings in \u03b3\u03b4 thymocytes + \u03b3\u03b4 T cells produce significantly more IL-4 and IL-5 after PMA/Ionomycin treatment compared with V\u03b34+ \u03b3\u03b4 T cells. This finding is consistent with earlier reports showing V\u03b31+ \u03b3\u03b4 T cells preferentially produce IL-4, and the depletion of V\u03b31+ subset cells increases host resistance against Listeria monocytogenes infection + \u03b3\u03b4 T cells suppress V\u03b34+ \u03b3\u03b4 T cell mediated antitumor function through IL-4 We identified 24 transcripts differentially expressed in resting V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells. We found Bclaf1 and Atf2 were preferentially expressed in V\u03b34+ \u03b3\u03b4 T cells while Hmga1 and Bcl11b were preferentially expressed in V\u03b31+ \u03b3\u03b4 T cells. As a transcriptional repressor, Bclaf1 interacts with several members of the Bcl2 protein family and plays a role in the regulation of apoptosis and DNA repair Bclaf1 also plays an important role in lymphocyte homeostasis and activation Atf2 transcription factor is a member of the leucine zipper family of DNA binding proteins and forms a homodimer or a heterodimer with c-Jun, stimulating cAMP responsive element (CRE) dependent transcription. Atf2 expression is lower in CD8+ T cells compared with CD4+ T cells, a functional explanation to the differential response to glucocorticoids between CD8+ and CD4+ T cells Hmga1 binds preferentially to the minor groove of AT rich regions in double stranded DNA. It is involved in many cellular processes including regulation of inducible gene transcription, insulin resistance, diabetes and malignant transformation Hmga1 in transcriptional silencing in T cell lineages and leukemic cells Bclaf1, Atf2 and Hmga1 in \u03b3\u03b4 T cells have not been reported. Bcl11b is a T-cell specific gene and required for T-lineage commitment. Aberrant expression of Bcl11b contributes to human T-ALL Bcl11b was preferentially expressed in V\u03b34+ \u03b3\u03b4 thymocytes Bcl11b preferentially expressed in activated V\u03b31+ \u03b3\u03b4 T cells. The role of the transcript isoform of Bcl11b in V\u03b31+ \u03b3\u03b4 T cells needs further study.Alternative splicing plays an important role in increasing functional diversity of eukaryotes. Compared with the ImmGen Project, one of the advantages of RNA-seq is able to quantify individual transcript isoforms and identify differentially expressed transcripts between V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells shared similar signaling pathways. We found higher expression of IL-4 and IL-5 in activated V\u03b31+ \u03b3\u03b4 T cells. This suggests a role in asthma given that V\u03b34+ \u03b3\u03b4 T cells suppress airway hyperresponsiveness, compared with V\u03b31+ \u03b3\u03b4 T cells that enhance airway hyperresponsiveness and raise levels of Th2 cytokines and eosinophils infiltration in the airways Many of the differentially expressed gene transcripts identified in activated V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells exhibited high levels of transcripts for several chemokines and chemokine receptors, including CCL4, CCL5, CCR2, CCR5 and CXCR3. These data highlight the role of V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells in immunoregulatory and inflammatory processes. For example, CCL4 (MIP-1beta) and CCL5 (RANTES) are both Th1-associated chemokines that bind to CCR5. Up-regulation of CCR5 ligands may play a role in the recruitment process of blood monocytes, memory T helper cells and eosinophils. CCR2 is expressed on both V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells, and is necessary for the accumulation of \u03b3\u03b4 TILs to the tumor bed + and V\u03b34+ \u03b3\u03b4 T cells. CXCR6 plays a critical role in NK cell memory of haptens and viruses + and V\u03b34+ \u03b3\u03b4 T memory cells needs further examination.Both resting V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells. For example, Itgae (Cd103), implicated in epithelial T cell retention, is highly expressed on V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells Itgae contributes to clustering and activation of V\u03b3 5 TCRs expressed by epidermal T cells Integrins play key roles in immune responses, leukocyte trafficking and many human diseases. Most integrin related research has been focused on \u03b1\u03b2 T cells, with little published on \u03b3\u03b4 T cells. Our results show several integrins were highly expressed in V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells. We show after PMA/Ionomycin treatment several activation markers of T cells were upregulated including CD25, CD69 and CD44, along with most cytokine genes in both subsets. In addition, activated V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells produced high levels of XCL1, CCL3, CCL4, CCL1, IFN-\u03b3, TNF\u03b1, Lta, Csf2 and IL-10. IFN-\u03b3 and TNF\u03b1 are Th1 type cytokines. Previous reports show V\u03b34+ \u03b3\u03b4 T cells are the major \u03b3\u03b4 T subset producing IFN-\u03b3, and they steer CD4+ T cells toward a dominant Th1 cell response + \u03b3\u03b4 T cells produced significantly more IFN-\u03b3 compared with V\u03b31+ \u03b3\u03b4 T cells, partly due to the high expression level of eomesodermin + \u03b3\u03b4 T cells were the major \u03b3\u03b4 T subset producing IFN-\u03b3 in response to L. monocytogenes infection + and V\u03b34+ \u03b3\u03b4 T cells produce high levels of IL-10.PMA/Ionomycin induces a robust non-TCR mediated response in V\u03b31+ \u03b3\u03b4 thymocytes expressed high levels of Stat4, Maf, Gata3 and Eomes compared with V\u03b31+ \u03b3\u03b4 thymocytes + and V\u03b34+ \u03b3\u03b4 T cells expressed high levels of these transcription factors and the levels of Gata3 were slightly higher in V\u03b31+ \u03b3\u03b4 T cells compared with V\u03b34+ \u03b3\u03b4 T cells after PMA/Ionomycin treatment. Gata3 is critical for Th2 cell differentiation and required for IL-4 production. The higher level of Gata3 expression in V\u03b31+ \u03b3\u03b4 T cells is consistent with the phenotype of V\u03b31+ \u03b3\u03b4 T cells producing more IL-4 than V\u03b34+ \u03b3\u03b4 T cells. T-bet is a major factor for Th1 cell differentiation and IFN-\u03b3 production Eomes is also involved in Th1 differentiation and IFN-\u03b3 production T-bet and Eomes is consistent with the phenotype of both V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells that produce high levels of IFN-\u03b3. The difference between our results with the ImmGen Project may be due to the source of \u03b3\u03b4 T cells. The cells used in the ImmGen Project are \u03b3\u03b4 thymocytes, however the cells in our study were peripheral \u03b3\u03b4 T cells derived from the spleen.Narayan et al. reported that V\u03b34+ and V\u03b34+ \u03b3\u03b4 T cells maintain inflammatory and regulatory phenotypes. Both demonstrate an inflammatory cell phenotype via IFN-\u03b3 and TNF\u03b1 expression. And, both display a regulatory cell phenotype via TGF-\u03b2 and IL-10 production. V\u03b31+ \u03b3\u03b4 T cells produced more Th2 type cytokines, while V\u03b34+ \u03b3\u03b4 T cells tended to produce more IL-17. Thus, Th2 type cytokines may explain how V\u03b31+ \u03b3\u03b4 T cells affect anti-inflammatory functions in different infection models, and describe the enhancing effect on airway hyperresponsiveness (AHR) + \u03b3\u03b4 T cells in the infection models and the inhibitory effect on airway hyperresponsiveness (AHR). Although this study was performed in V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells expanded in vitro, which may not fully represent the true status of V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells in vivo, our results support the hypothesis that distinct \u03b3\u03b4 TCR types direct cells to acquire a certain type of functional programming during thymic development Taken together, this study shows both V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells following PMA/Ionomycin treatment. Although both \u03b3\u03b4 T cell populations have similar transcript profiles, subset-specific transcripts define distinct characteristics and describe the inherent differences between V\u03b31+ and V\u03b34+ \u03b3\u03b4 T cells.Complementary to the ImmGen Project, this report provides a comprehensive gene expression profile of mouse peripheral V\u03b31Dataset S1Raw data and differential expression analysis in RNA-seq.(XLSX)Click here for additional data file.Dataset S2+ \u03b3\u03b4 T cells.Differentially expressed genes between the resting and activated V\u03b31(XLSX)Click here for additional data file.Dataset S3+ \u03b3\u03b4 T cells.Differentially expressed genes between the resting and activated V\u03b34(XLSX)Click here for additional data file.Dataset S4Transcription factors related to Th cell differentiation and cytokine secretion.(XLSX)Click here for additional data file."} +{"text": "The packing features supra\u00admolecular layers sustained by O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonding.A distorted square-pyramidal CdNS 6H12NOS2)2(C4H10N2)], features a distorted square-pyramidal coordination geometry about the central CdII atom. The di\u00adthio\u00adcarbamate ligands are chelating, forming similar Cd\u2014S bond lengths and define the approximate basal plane. One of the N atoms of the piperazine mol\u00adecule, which adopts a chair conformation, occupies the apical site. In the crystal, supra\u00admolecular layers propagating in the ac plane are formed via hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy), hy\u00addroxy-O\u2014H\u22efN and coordinated-piperazine-N\u2014H\u22efO(hy\u00addroxy) hydrogen bonds; the layers also feature methine-C\u2014H\u22efS inter\u00adactions and S\u22efS [3.3714\u2005(10)\u2005\u00c5] short contacts. The layers stack along the b-axis direction with very weak terminal-piperazine-N\u2014H\u22efO(hy\u00addroxy) inter\u00adactions between them. An evaluation of the Hirshfeld surfaces confirms the importance of inter\u00admolecular inter\u00adactions involving oxygen and sulfur atoms.The title compound, [Cd(C Layers thus formed stack along the b axis, with very weak terminal-piperazine-N4\u2014H\u22efO2(hy\u00addroxy) inter\u00adactions between them, Table\u00a02In the extended structure, hy\u00addroxy-O1\u2014H\u22efO2(hy\u00addroxy), hy\u00addroxy-O2\u2014H\u22efN4 and (coordinated-piperazine)-N3\u2014H\u22efO1(hy\u00addroxy) hydrogen bonds Table\u00a02 lead to Crystal Explorer , and there is no contribution from C\u22efC contacts in the structure as the result of the absence of C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions. Finally, the presence of S\u22efS contacts can also be viewed in the delineated fingerprint plot, Fig.\u00a07g, by the density of points in the ed, id region around 1.7\u20132.4\u2005\u00c5 as a broken line segment.The overall two-dimensional fingerprint plot, Fig.\u00a072% Fig.\u00a07d, and tet al., 2014The identified inter\u00admolecular inter\u00adactions were further evaluated by an analysis of the enrichment ratios (ER) that give a qu\u00adanti\u00adtative measure of the likelihood of specific inter\u00admolecular inter\u00adactions to occur based on a Hirshfeld surface analysis 2]2 compounds are usually binuclear CH2CH2OH]2 CH2CH2OH]2\u00b7EtOH]x for x = 2 and, unprecedented, n form transforming in solution to the thermodynamic, binuclear form (x = 2). Other recrystallization conditions led to decomposition of the di\u00adthio\u00adcarbamate ligands and subsequent formation of a co-crystal and some salts. This behaviour, along with the unexpected structure of {Cd[S2CN(iPr)CH2CH2OH]22] 2(MeOH)2 CH2CH2OH]2 : \u03bd (O\u2014H) 3281, \u03bd(C\u2014N) 1442, \u03bd (C\u2014O) 1169, \u03bd (C\u2014S) 1029.The reagents Cd[SUiso(H) set to 1.2Ueq(C). The oxygen- and nitro\u00adgen-bound H atoms were located in a difference Fourier map but were refined with a distance restraints of O\u2014H = 0.84\u00b10.01\u2005\u00c5 and N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.5Ueq(O) and 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989016000165/hb7558sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016000165/hb7558Isup2.hklStructure factors: contains datablock(s) I. DOI: 1445316CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In three potentially anti\u00adconvulsant compounds, of which two are isoindoline derivatives and one an iso\u00adquinoline derivative, the central moiety is planar. In the crystals of all three compounds, there are C\u2014H\u22efO hydrogen bonds present linking the mol\u00adecules into two-dimensional slabs for the isoindoline derivatives, and into a three-dimensional framework for the iso\u00adquinoline derivative. 9H7NO3, (1), C10H7NO5, (2), and C14H9NO5, (3), are three potentially anti\u00adconvulsant compounds. Compounds (1) and (2) are isoindoline derivatives and (3) is an iso\u00adquinoline derivative. Compounds (2) and (3) crystallize with two independent mol\u00adecules (A and B) in their asymmetric units. In all three cases, the isoindoline and benzoiso\u00adquinoline moieties are planar . The substituents attached to the N atom are almost perpendicular to the mean planes of the heterocycles, with dihedral angles of 89.7\u2005(3)\u00b0 for the N\u2014O\u2014Cmeth\u00adyl group in (1), 71.01\u2005(4) and 80.00\u2005(4)\u00b0 for the N\u2014O\u2014C(=O)O\u2014Cmeth\u00adyl groups in (2), and 75.62\u2005(14) and 74.13\u2005(4)\u00b0 for the same groups in (3). In the crystal of (1), there are unusual inter\u00admolecular C=O\u22efC contacts of 2.794\u2005(1) and 2.873\u2005(1)\u2005\u00c5 present in mol\u00adecules A and B, respectively. There are also C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [inter-centroid distance = 3.407\u2005(3)\u2005\u00c5] present, forming slabs lying parallel to (001). In the crystal of (2), the A and B mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, forming slabs parallel to (10-1), which are in turn linked via a number of \u03c0\u2013\u03c0 inter\u00adactions [the most significant centroid\u2013centroid distances are 3.4202\u2005(7) and 3.5445\u2005(7)\u2005\u00c5], forming a three-dimensional structure. In the crystal of (3), the A and B mol\u00adecules are linked via C\u2014H\u22efO hydrogen bonds, forming a three-dimensional structure, which is consolidated by \u03c0\u2013\u03c0 inter\u00adactions [the most significant inter-centroid distances are 3.575\u2005(3) and 3.578\u2005(3)\u2005\u00c5].The title compounds, C The compound is a dual MES/6Hz active compound. Compounds (2) and (3) showed similar activity.2-Meth\u00adoxy\u00adisoindoline-1,3-dione, (1), studied by X-ray techniques, was inactive in MES and scPTZ in mice, but showed MES protection in rat studies at 50\u2005mg\u2005kgThe title compounds, containing either an isoindoline-1,3-dione moiety, (1) Fig.\u00a01 and (2) In compound (1), the isoindoline ring is planar [r.m.s. deviation = 0.017\u2005(4)\u2005\u00c5]. The meth\u00adoxy O atom, O3, deviates from this plane by 0.176\u2005(6)\u2005\u00c5 while the methyl C atom, C9, is out of the plane by 1.105\u2005(9)\u2005\u00c5. The meth\u00adoxy substituent is oriented almost perpendicular to the indoline ring with the dihedral angle between the mean planes of the indoline ring and the meth\u00adoxy substituent being 89.7\u2005(3)\u00b0.A and B) in the asymmetric unit. The isoindoline ring is planar [r.m.s. deviation = 0.0327\u2005(9) for A and 0.0147\u2005(9)\u2005\u00c5 for B] with the dione O atoms significantly out of the plane for mol\u00adecule A but not for mol\u00adecule B . The carbonato moiety is planar in both mol\u00adecules and makes dihedral angles of 71.50\u2005(3) and 80.03\u2005(4)\u00b0 with the benzoiso\u00adquinoline ring in A and B, respectively, indicating that these substituents are oriented almost perpendicular to the benzoiso\u00adquinoline ring system.In compound (2), there are two mol\u00adecules (A and B) in the asymmetric unit. In both mol\u00adecules, the benzoiso\u00adquinoline ring systems are planar . The meth\u00adoxy O atom deviates from this plane by 0.126\u2005(1) for atom O5A in A and 0.156\u2005(1)\u2005\u00c5 for atom O5B in B. The methyl carbonate moieties are planar and these substituents are oriented almost perpendicular to the iso\u00adquinoline rings, making dihedral angles of 71.50\u2005(3) and 80.04\u2005(4)\u00b0 for A and B, respectively. As in (2), these dihedral angles are significantly smaller than that found for (1).In compound (3), there are also two mol\u00adecules \u2005\u00c5; Cg1 and Cg2 are the centroids of rings N1/C1/C2/C7/C8 and C2\u2013C7, respectively; symmetry codes: (i) x\u00a0\u2212\u00a01, y, z; (ii) x\u00a0+\u00a01, y, z].In the crystal of (1), there are C\u2014H\u22efO hydrogen bonds Fig.\u00a04 and \u03c0\u2013\u03c0 A and B mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds for Cg1\u22efCg5ii and 3.5445\u2005(7)\u2005\u00c5 for Cg2\u22efCg4ii .In the crystal of (2), the ds Fig.\u00a05, formingA and B mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds , Cg2\u22efCg3iii = 3.575\u2005(3)\u2005\u00c5 and Cg9\u22efCg10iv; Cg1, Cg2, Cg3, Cg9 and Cg10 are the centroids of rings N1A/C1A\u2013C5A, C2A/C3A/C6A\u2013C9A, C3A/C4A/C9A\u2013C12A, C2B/C3B/C6B\u2013C9B and C3B/C4B/C9B\u2013C12B, respectively; symmetry codes: (iii) x, \u2212y\u00a0+\u00a0z\u00a0\u2212\u00a0x, \u2212y\u00a0+\u00a0z\u00a0+\u00a0In the crystal of (3), the ds Fig.\u00a06, formingA and O1B) has a short inter\u00admolecular inter\u00adactions with the central C atom of the carbonato group , which is perpendicular to the carbonato plane indicating that both atoms, C9A and C9B, must have significant positive character. These inter\u00adactions link the mol\u00adecules into dimers as shown in Figs. 6A (but not for mol\u00adecule B), where a longer inter\u00admolecular inter\u00adaction of 3.060\u2005(3)\u2005\u00c5 is observed between atoms O2A and C13A, resulting in weakly associated dimers similar to that seen in the case of (2).Inter\u00adestingly, in the crystal of (2) one of the two dione moieties for each mol\u00adecule :N-hy\u00addroxy\u00adphthalimide in absolute ethanol (350\u2005ml), and the red reaction mixture was stirred at room temperature for 30\u2005min. The brick-red precipitate was collected, washed with water, and dried in the oven at 373\u2005K for 30\u2005min to give 17.45\u2005g (95%) of sodium phthalimide oxide as brick-red crystals; m.p. > 573\u2005K. To the solution of sodium phthalimide oxide in water (15\u2005ml) was added acetone (10\u2005ml), followed by a solution of bromo\u00admethane . The reaction mixture was stirred at room temperature for 16\u2005h, during which the red color disappeared. On standing at room temperature for 48\u2005h, the product solidified in the aqueous mixture and was collected. Recrystallization from 2-propanol gave 0.72\u2005g (78%) of compound (1) as plate-like colorless crystals: m.p. 395\u2013397\u2005K; 1H NMR (CDC13) \u03b4 3.36 , 5.52, s, 1 H,CH, 7.87 .To a freshly prepared solution of sodium in absolute ethanol (60\u2005ml) was added a solution of Compound (2):1H NMR (CDC13) \u03b4 3.8 , 7.86 .To a solution of sodium phthalimide oxide in water (15\u2005ml) was added acetone (10\u2005ml), followed by a solution of bromo\u00ad(meth\u00adoxy)methanone . The reaction mixture was stirred at room temperature for 16\u2005h, during which the red color disappeared. On standing at room temperature for 48\u2005h, the product solidified in the aqueous mixture and was collected. Recrystallization from ethanol gave 0.82\u2005g (74%) of compound (2) as colorless crystals: m.p. 410\u2013411\u2005K; Compound (3):1H NMR (CDCl3) \u03b4 3.79 , 5.66 , 7.65\u20138.50 .To a solution of sodium naphthalimide oxide, , in water (50\u2005ml), was added bromo\u00ad(meth\u00adoxy)methanone in acetone (10\u2005ml). The red reaction mixture was stirred at room temperature. The red color disappeared within 5\u2005min and the reaction mixture was filled with a white precipitate. After standing for 4\u2005h, the white precipitate was collected, washed with water, and recrystallized from ethanol to give 1.46\u2005g (89%) of compound (3) as colorless crystals: m.p. 483\u2013485\u2005K; isoU(H) = 1.5Ueq(C) for methyl H atoms and = 1.2eqU(C) for other H atoms.Crystal data, data collection and structure refinement details for (1), (2) and (3) are summarized in Table\u00a0410.1107/S1600536814023769/su2795sup1.cifCrystal structure: contains datablock(s) 1, 2, 3. DOI: 10.1107/S1600536814023769/su27951sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S1600536814023769/su27952sup3.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S1600536814023769/su27953sup4.hklStructure factors: contains datablock(s) 3. DOI: Click here for additional data file.10.1107/S1600536814023769/su27951sup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S1600536814023769/su27952sup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S1600536814023769/su27953sup7.cmlSupporting information file. DOI: 1031391, 1031392, 1031393CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound, bis\u00ad\u00adcobalt(III) tris\u00ad(perchlorate) methanol monosolvate monohydrate, is a novel complex in which the metal centre is hexa\u00adcoordinated. 25H30N4O4)2](ClO4)3\u00b7CH3OH\u00b7H2O, the metal atom is coordinated by two tridentate crown ether terpyridine ligands, forming a distorted CoN6 octa\u00adhedron. The three pyridine rings in each crown\u2013terpyridine ligand are approximately coplanar [maximum deviations = 0.088\u2005(12) and 0.102\u2005(15)\u2005\u00c5] and the mean planes through the three pyridine rings are perpendicular to each other, making a dihedral angle of 89.95\u2005(17)\u00b0. An intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction is observed between the two terpyridine ligands. In the crystal, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, a \u03c0\u2013\u03c0 stacking inter\u00adaction [centroid\u2013centroid distance = 3.923\u2005(7)\u2005\u00c5] and a C\u2014H\u22ef\u03c0 inter\u00adaction connect the complex cation, the perchlorate anions and the two types of solvent molecules, forming a three-dimensional network.In the title compound, [Co(C M(R-terpy)]X2 , have been investigated because of their inter\u00adesting properties such as magnetic and photochemical characteristics. Cobalt(II) complexes with R-terpy ligands are known as spin-crossover compounds. Previously, we observed the unique spin-transition behavior in [Co(II)(R-terpy)2](BF4)2 with long-alkyl\u00adated terpyridine ligands, and showed that the magnetic behaviors are influenced not only by inter-chain inter\u00adactions between long alkyl chains but also by \u03c0\u2013\u03c0 stacking inter\u00adactions between terpyridine moieties compounds. The Co\u2014N distances of the central pyridine in the terpyridine unit [1.844\u2005(9)\u2005\u00c5] are shorter than the Co\u2014N distances of the side pyridine in the terpyridine unit [1.910\u2005(10)\u20131.949\u2005(10)\u2005\u00c5], which induces a pronounced distortion of the CoN6 octa\u00adhedron. The three pyridine rings in each crown\u2013terpy ligand are approximately coplanar [maximum deviations 0.102\u2005(15) and 0.088\u2005(12)\u2005\u00c5], and the two mean planes through the three pyridine rings in the complex are nearly perpendicular to each other, making a dihedral angle of 89.95\u2005(17)\u00b0.The asymmetric unit of the title compound consists of one [Co(crown\u2013terpy)The overall packing of structure is shown in Fig.\u00a024)3 dissolved in methanol (20\u2005ml) was poured dropwise into a solution of the crown\u2013terpy ligand in 1:1 methanol\u2013chloro\u00adform. The precipitate formed immediately and was filtered. Single crystals of the title compound suitable for X-ray diffraction were obtained from a methanol solution.The crown-terpyridine ligand was prepared by the reaction of 4\u2032-bromo-2,2\u2032:6\u2032,2\u2032\u2032-terpyridine and 1,4,7,10-tetra\u00adoxa-13-aza\u00adcyclo\u00adpenta\u00addecane in DMF. The mixed solution was evaporated to give the ligand as a white powder. Co(ClOUiso(H) = 1.2Ueq(C) and 1.5Ueq. The positions of the H atoms of the water mol\u00adecule were refined with restraints of O\u2014H = 0.85\u2005(2) and H\u22efH = 1.38\u2005(2)\u2005\u00c5, and with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015014164/is5400sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015014164/is5400Isup2.hklStructure factors: contains datablock(s) I. DOI: 1415327CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title complex is a centrosymmetric dimer with a copper\u2013copper distance of 4.0408\u2005(3)\u2005\u00c5. The Cu ions in the dimer are bridged by two triazole rings and oxygen donor ligands from water mol\u00adecules and nitrate anions in a distorted octa\u00adhedral coordination geometry. 2(C10H8N7)2(NO3)2(H2O)2]\u00b72H2O, consists of centrosymmetric dimeric units with a copper\u2013copper separation of 4.0408\u2005(3)\u2005\u00c5. The CuII ions in the dimer display a distorted octa\u00adhedral coordination geometry and are bridged by two triazole rings, forming an approximately planar Cu2N4 core (r.m.s. deviation = 0.049\u2005\u00c5). In the crystal, O\u2014H\u22efO, O\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions link the mol\u00adecules into a three-dimensional network.The structure of the dinuclear title complex, [Cu The study of this type of coordination compound is promising since, as a result, a compound can be obtained with useful physical properties such as optical, magnetic or catalytic \u2005\u00c5 for atom N2], with a Cu\u22efCu separation of 4.0408\u2005(3)\u2005\u00c5, in good agreement with the values usually observed in \u03bc-triazolyl-bridged complexes \u2005\u00c5; \u03b82 = 88.62\u2005(16)\u00b0], while the five-membered Cu1/N2/C1/C2/N4 chelate ring adopts a flattened envelope conformation with the Cu atom as flap .The structure of the title complex mol\u00adecule Fig.\u00a01 has a crCg1\u22efCg2ii = 3.8296\u2005(13)\u2005\u00c5 and Cg3\u22efCg3iii = 3.5372\u2005(10), and perpendic\u00adular inter\u00adplanar distances Cg1\u22efCg2ii = 3.5584\u2005(9) and Cg3\u22efCg3iii = 3.3234\u2005(10)\u2005\u00c5 .In the crystal, the complex molecules and water mol\u00adecules of crystallization are linked through O\u2014H\u22efO, O\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds Table\u00a01, forming2\u2013Cu. The most similar are: di\u00adaqua\u00adbis\u00ad-1,2,4-triazolato-N\u2032,N1,N2,N\u2032\u2032)bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfonato-O)dicopper(II) -1,2,4-triazolato]di\u00adaqua\u00addicopper diperchlorate -5-(meth\u00adyl)-1,2,4-triazol-1-ide]tri\u00adaqua\u00adtricopper di\u00adperchlorate dihydrate bis(acetonitrile)\u00adbis\u00ad(perchlorato-O)dicopper 2\u00b73H2O was added to a hot solution of 2-[5--triazol-1-yl-methyl-1H--triazol-3\u00adyl]pyridine in water (7\u2005ml). The transparent blue solution was left to evaporate slowly in the air and after few hours, blue single crystals suitable for X-ray analysis were obtained (yield: 67%).A water solution of Cu(NOUiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016003479/rz5185sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016003479/rz5185Isup2.hklStructure factors: contains datablock(s) I. DOI: 1456451CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The aza\u00adadamantane structure in (I) is slightly distorted, with N\u2014CH2\u2014CH2\u2014N torsion angles of 10.4\u2005(3) and \u22129.0\u2005(3)\u00b0. These values differ slightly from the corresponding torsion angles in the free aminal cage (0.0\u00b0) and in related co-crystalline adducts, which are not far from a planar geometry and consistent with a Dd2 mol\u00adecular symmetry in the tetra\u00adaza\u00adtri\u00adcyclo structure. The structures also differ in that there is a slight elongation of the N\u2014C bond lengths about the N atom that accepts the hydrogen bond in (I) compared with the other N\u2014C bond lengths. In the crystal, the two mol\u00adecules are not only linked by a classical O\u2014H\u22efN hydrogen bond but are further connected by weak C\u2014H\u22ef\u03c0 inter\u00adactions, forming a two-dimensional supra\u00admolecular network parallel to the bc plane.In the crystal of the title co-crystalline adduct, C Reaction between TATD with 4-chloro-3,5-di\u00admethyl\u00adphenol in solution yields symmetrical bis-benzoxazines -1,8,10,12-tetra\u00adaza\u00adtetra\u00adcyclo\u00adpenta\u00addecane with 4-nitro\u00adphenol -4-bromo\u00adphenol \u20134-chloro-3,5-di\u00admethyl\u00adphenol under solvent-free conditions by simply grinding together the components at room temperature. Herein, we describe the synthesis of the title co-crystalline adduct 1,3,6,8-tetra\u00adaza\u00adtri\u00adcyclo\u00addodecane (TATD) and a 4-chloro-3,5-di\u00admethyl\u00adphenol mol\u00adecule linked via an O\u2014H\u22efN hydrogen bond, forming a D motif ; p-bromo\u00adphenol (pKa = 9.37) and hydro\u00adquinone (pKa = 9.85) et al., 2007et al., 2015et al., 1987et al., 20142dD point group, two small differences are noted. The aza\u00adadamantane structure in (I)2\u2014CH2\u2014N torsion angles of 10.4\u2005(3)\u00b0 (N1\u2014C1\u2014C2\u2014N2) and \u22129.0\u2005(3)\u00b0 (N3\u2014C7\u2014C8\u2014N4). These values differ slightly from the values of the corresponding torsion angles in the free aminal cage and 1.480\u2005(2)\u2005\u00c5], compared to the the other N\u2014C bond lengths \u2005\u00c5; symmetry operator: (i) x, \u2212y, z\u00a0\u2212\u00a0c-axis direction e Table\u00a02. Furtheron Fig.\u00a03.meth\u00adyl = 1.502\u2005(3), 1.500\u2005(3), 1.514\u2005(3) and 1.505\u2005(3)\u2005\u00c5]. For 1,3,6,8-tetra\u00adaza\u00adtri\u00adcyclo\u00addodecane, two comparable structures were retrieved from the CSD 3,8]dodecane co-crystallized with hydro\u00adquinone 3,8]dodecane is rather limited.The geometric parameters of 4-chloro-3,5-di\u00admethyl\u00adphenol in (I)3,8]dodecane (TATD) and 4-chloro-3,5-di\u00admethyl\u00adphenol was ground using a mortar and pestle, at room temperature for 15\u2005min., as required to complete the reaction (TLC). The mixture was then dissolved in methanol. Crystals suitable for X-ray diffraction were obtained from a methanol solution upon slow evaporation of the solvent at room temperature.A mixture of 1,3,6,8-tetra\u00adaza\u00adtri\u00adcyclo\u00ad[4.4.1.Uiso(H) values set at 1.2Ueq (1.5 for methyl groups) of the parent atom. The methyl groups were allowed to rotate but not to tip.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015010257/zs2335sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015010257/zs2335Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015010257/zs2335Isup3.cmlSupporting information file. DOI: 1403518CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Fe\u2014N bond lengths are indicative of a d6 low-spin state for the FeII ion in the complex. The dihedral angle between the phen ligands in the cation is 87.64\u2005(6)\u00b0. The Tf2N\u2212 counter-anion is non-coordinating, with the \u2013CF3 groups arranged in a trans fashion with respect to each other, leading to an anti,anti conformation of the \u2013CF3 groups and \u2013SO2N\u2013 moieties relative to the S\u2014C bonds. The water mol\u00adecule of crystallization connects two O atoms of the Tf2N\u2212 anions through weak hydrogen bonds. C\u2014H\u22efO hydrogen-bonding inter\u00adactions are also observed, consolidating the packing of the mol\u00adecules into a three-dimensional network structure.The crystal structure of the title complex, [Fe(C DOI: 10.1107/S2056989014026966/wm5100Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989014026966/wm5100fig1.tif3 2+ 2 \u2212 2 x y z . DOI: 3]2+, Tf2N\u2212 and H2O mol\u00adecular units. Displacement ellipsoids are represented at the 30% probability level. Hydrogen atoms were omitted for clarity. [Symmetry code: i) \u2212x\u00a0+\u00a01,y,-z\u00a0+\u00a0View of the [Fe(phen)Click here for additional data file.10.1107/S2056989014026966/wm5100fig2.tif2 2 \u2212 x y z x y z . DOI: 2O mol\u00adecule and Tf2N\u2212 anions. [Symmetry codes: ii) x\u00a0\u2212\u00a0y\u00a0\u2212\u00a01/2,z, iii) \u2212x\u00a0+\u00a0y\u00a0\u2212\u00a0z\u00a0+\u00a0Hydrogen bonds between the H1038289CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The cationic NiII center is in a distorted square-planar coordination environment chelated by the imine N and phenolate O donor atoms of the two Schiff base ligands. The N and O donor atoms of the two ligands are mutually trans with Ni\u2014N and Ni\u2014O bond lengths of 1.9242\u2005(10) and 1.8336\u2005(9)\u2005\u00c5, respectively. The fluoro\u00adphenyl ring is almost orthogonal to the coordination plane and makes a dihedral angle of 82.98\u2005(7)\u00b0 with the phenolate ring. In the crystal, mol\u00adecules are linked into screw chains by weak C\u2014H\u22efF hydrogen bonds. Additional C\u2014H\u22ef\u03c0 contacts arrange the mol\u00adecules into sheets parallel to the ac plane.The asymmetric unit of the title complex, [Ni(C In particular they can be photoluminescent -(4-fluoro\u00adbenz\u00adyl)imino\u00admeth\u00adyl]-6-meth\u00adoxy\u00adphenolato-\u03ba2N,O1}nickel(II) -(4-meth\u00adoxy\u00adbenz\u00adyl)imino\u00admeth\u00adyl]phenolato-\u03ba2N,O1}nickel(II) 2] (1), and its characterization by spectros\u00adcopy and elemental analysis. Crystal structure determination confirms the binding mode of the [(4-fluoro\u00adbenz\u00adyl)imino\u00admeth\u00adyl]phenolate ligand to the NiII cation \u00b0 and O1\u2014Ni1\u2014N1i = 87.44\u2005(4)\u00b0 . As expected under inversion symmetry, the trans angles (N11\u2014Ni1\u2014N1i and O1\u2014Ni1\u2014O1i) are found to be linear. The Ni1\u2014N1 and Ni1\u2014O1 distances in the N2O2 coordination plane are 1.9242\u2005(10)\u2005\u00c5 and 1.8336\u2005(9)\u2005\u00c5, respectively. These compare well with those observed in the two other closely related NiII complexes with N2O2 coordinating Schiff base ligands \u2005\u00c5, \u03b8 = 65.3\u2005(3) and \u03d5 = 4.0\u2005(3)\u00b0. Other bond lengths and angles observed in the structure are also normal. The fluoro\u00adphenyl ring (C9\u2013C14) makes a dihedral angle of 82.98\u2005(7)\u00b0 with the phenolate ring (C1\u2013C6).The asymmetric unit of (1) contains one-half of the mol\u00adecule with the Nind Fig.\u00a01. The catA\u22efF1 inter\u00adactions phenolate residues. No corresponding structures with a benzyl or substituted benzyl unit bound to the imino N atom were found. However extending the search to allow additional substitution on the phenolate ring resulted in eight discrete structures including the two closely related structures mentioned previously revealed a total of 1191 NiE)-2-[(4-fluoro\u00adbenzyl\u00adimino)\u00admeth\u00adyl]phenol. Nickel(II) acetate tetra\u00adhydrate was dissolved separately in absolute ethanol (10\u2005ml) and added to a flask containing the cooled ligand solution. The mixture was stirred and refluxed for 3\u2005h upon which a dark-green solid formed. This was filtered off, washed with ice-cold ethanol and air-dried at room temperature. The solid product was recrystallized from chloro\u00adform, yielding green crystals. Yield 68.6%; m.p. 471\u2013473\u2005K. Analytical data for C28H22F2N2O2Ni: C, 65.28; H, 4.30; N, 5.44. Found: C, 65.87; H, 4.39; N, 5.55. IR : \u03bd(C=N) 1612 (s), \u03bd(C\u2014N) 1390 (w), \u03bd(C\u2014O) 1221 (s), \u03bd(Ni\u2014N) 597 (w), \u03bd(Ni\u2014O) 451 (w). The infrared spectra of the title complex revealed a strong band of 1612\u2005cm\u22121 in the spectrum assignable to C=N stretching frequency upon complexation was added to salicyl\u00adaldehyde , dissolved in absolute ethanol (2\u2005ml), forming a bright-yellow solution. The mixture was heated under reflux for an hour to produce the ligand, = 0.95\u2005\u00c5 for aromatic and 0.99\u2005\u00c5 for CH2 hydrogen atoms. The Uiso values were constrained to be 1.2Ueq of the carrier atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814020546/sj5425sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S1600536814020546/sj5425Isup2.hklStructure factors: contains datablock(s) I. DOI: 1024161CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efF inter\u00adactions connect the mol\u00adecules, forming supra\u00admolecular chains propagating along the 22H23F2N3O2, the bicyclic ring system exists in a twin-chair conformation with an equatorial disposition of the 4-fluoro\u00adphenyl groups on the heterocycle. These aromatic rings are inclined to one another by 19.4\u2005(1)\u00b0. In the crystal, mol\u00adecules are linked by pairs of N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds into inversion dimers, incorporating R12(7) and R22(8) ring motifs; the same O atom accepts both hydrogen bonds. These dimers are further linked by a pair of C\u2014H\u22efF hydrogen bonds, enclosing R22(28) ring motifs, forming supra\u00admolecular chains along [010]. The NH group of the pyridine ring is not involved in hydrogen bonding, probably due to the steric hindrance of the fluoro\u00adphenyl groups.In the title compound, C The fluoro\u00adphenyl groups on the heterocycle occupy equatorial positions and are inclined to one another by 19.4\u2005(1)\u00b0. The geometric parameters of the title mol\u00adecule agree well with those reported for similar structures, for example, 2,4-bis\u00ad(4-fluoro\u00adphen\u00adyl)-3-aza\u00adbicyclo\u00ad[3.3.1]nonan-9-one, (II) i and C2\u2014H2\u22efO3i .In the crystal, pairs of bifurcated acceptor N3\u2014H3\u22efO3i Table\u00a01 hydrogenfs Fig.\u00a02. These di Table\u00a01 hydrogenviz. compound (III) in the asymmetric unit. In all three compounds, the bi\u00adcyclo rings have twin-chair conformations with equatorially disposed 4-fluoro\u00adphenyl groups on the heterocycle. The fluoro\u00adphenyl rings are oriented at an angle of 28.7\u2005(1)\u00b0 in (II), and 55.3\u2005(1) (mol\u00adecule A) and 56.4\u2005(1)\u00b0 (mol\u00adecule B) for (III), compared to 19.4\u2005(1)\u00b0 in the title compound, (I)38 \u2018hits\u2019 for crystal structures containing the 3-aza\u00adbicyclo\u00ad[3.3.1]nonane subunit were obtained for a search of the Cambridge Structural Database medium, with the addition of few drops of acetic acid, was stirred for 10\u201312\u2005h. After completion of the reaction a solid mass was formed. The precipitate was filtered off and washed with an ethanol\u2013water mixture. The crude product was then recrystallized from ethanol\u2013chloro\u00adform to obtain colourless diffraction-quality crystals of title compound.A mixture of 2,4-diphenyl-3-aza\u00adbicyclo\u00ad[3.3.1]nonan-9-one (0.1\u2005mmol), methyl hydrazine\u00adcarboxyl\u00adate (1.5\u2005mmol) in an ethanol\u2013chloro\u00adform (1:1 Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq for all other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814018935/su2773sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S1600536814018935/su2773Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814018935/su2773Isup3.cmlSupporting information file. DOI: 1020373CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the planes of the phen\u00adoxy and phthalo\u00adnitrile rings are oriented at a dihedral angle of 60.39\u2005(5)\u00b0. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds, forming slabs parallel to (100). The slabs are linked by a pair of inversion-related C\u2014H\u22efN hydrogen bonds, forming a three-dimensional structure. 20H20N4O2\u00b7H2O, the planes of the phen\u00adoxy and phthalo\u00adnitrile rings are oriented at a dihedral angle of 60.39\u2005(5)\u00b0. The 3-(di\u00ad\u00admethyl\u00adamino)\u00adpropyl chain has an extended conformation and is cis with respect to the phthalo\u00adnitrile ring. In the crystal, O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules to form slabs parallel to (100). There are also C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions present within the slabs. The slabs are linked by a pair of inversion-related C\u2014H\u22efN hydrogen bonds, involving phthalo\u00adnitrile rings, forming a three-dimensional structure.In the title compound, C Well-known application fields for amino derivatives are their use as synthetic inter\u00admediates of anti\u00adcancer agents, anti\u00adbiotics and other drugs. They also exhibit exceptionally low ocular irritation and oral toxicity, being well tolerated by human tissue , O\u2014Hw. .Oamd and O\u2014Hw\u22efNdma (dma = di\u00admethyl\u00adamino) hydrogen bonds , and the O\u2014Hw\u22ef Oamd, C\u2014Hphen\u22efOamd and C\u2014Hphen\u22efOw hydrogen bonds form via a pair of inversion-related Cphn\u2014H\u22efNphn hydrogen bonds, forming a three-dimensional structure phthalo\u00adnitrile \u00b0.A search of the Cambridge Structural Database = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for the other H atoms.The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a02N,N-di\u00admethyl\u00adpropane-1,3-di\u00adamine and K2CO3 in dry tetra\u00adhydro\u00adfuran , stirred in an ice bath for 15\u2005min, was added over a period of 40\u2005min, 4-benzoyl chloride in dry THF (5\u2005ml). The reaction mixture was then stirred for 5\u2005h at room temperature and monitored by thin-layer chromatography [THF\u2013hexane (3:4 v/v) as a mobile phase on silica-gel plates]. The oily residue obtained was dissolved in MeOH. The solvent was evaporated slowly and colourless block-like crystals appeared in ca 10\u2005d .To a mixture of 10.1107/S2056989015014991/su5187sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015014991/su5187Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015014991/su5187Isup3.cmlSupporting information file. DOI: 1418026CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "P,S-donor set is observed in the title polymorph; an intra\u00admolecular Au\u22efO inter\u00adaction is noted. The packing is consolidated by C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions to generate a three-dimensional network.A linear geometry defined by a 8H7ClNOS)(C18H15P)], is a monoclinic polymorph of the previously reported triclinic form [Tadbuppa & Tiekink coordination geometry [P\u2014Au\u2014S = 175.62\u2005(5)\u00b0 in the title polymorph], being coordinated by thiol\u00adate S and phosphane P atoms, a Z conformation about the C=N bond and an intra\u00admolecular Au\u22efO contact. The major conformational difference relates to the relative orientations of the residues about the Au\u2014S bond: the P\u2014Au\u2014S\u2014C torsion angles are \u22128.4\u2005(7) and 106.2\u2005(7)\u00b0 in forms \u03b1 and \u03b2, respectively. The mol\u00adecular packing of form \u03b2 features centrosymmetric aggregates sustained by aryl-C\u2014H\u22efO inter\u00adactions, which are connected into a three-dimensional network by aryl-C\u2014H\u22ef\u03c0 contacts. The Hirshfeld analysis of forms \u03b1 and \u03b2 shows many similarities with the notable exception of the influence of C\u2014H\u22efO inter\u00adactions in form \u03b2.The title compound, [Au(Ckink 2010. Acta Cr N-aryl-O-alkyl\u00adthio\u00adcarbamates, i.e. compounds of general formula R3PAu[SC(OR\u2032)=NR\u2032\u2032] continues owing to their recently disclosed exciting biological activities. Thus, various tri\u00adphenyl\u00adphosphane derivatives display excellent cytotoxicity profiles against HT-29 colon cancer cells, a particularly virulent form of cancer, and mechanistic studies have shown these to induce both intrinsic and extrinsic pathways of cell death leading to apoptosis et al., 2005et al., 2007i.e. 3.052\u2005(3)\u2005\u00c5. The pattern of bond angles about the quaternary carbon atom, C1, follow the expected trends with the widest angle involving the sulfur and doubly bonded nitro\u00adgen atom and with the narrowest angle involving the single-bonded atoms. The conformation about the formal C1=N1 bond, Table\u00a01Z.The mol\u00adecular structure of the new monoclinic form of (I)P21/n with Z\u2032 = 1. The earlier polymorph, by contrast, crystallizes in triclinic space group PZ\u2032 = 1. A comparison of the key geometric parameters is given in Table\u00a01Form \u03b2 crystallizes in the monoclinic space group a and Table\u00a02b and Table\u00a023P ligands align to form a so-called six-fold phenyl embrace (6PE) Supra\u00admolecular dimers feature in the mol\u00adecular packing of form \u03b2 of (I)i.e. forms \u03b1 and \u03b2, were studied through Hirshfeld surface analysis by mapping on the normalized contact distance (dnorm) upon computation of the inner (di) and outer (de) distances of the Hirshfeld surface to the nearest nucleus Supra\u00admolecular features, C\u2014H\u22ef\u03c0 inter\u00adactions feature in both structures. To a first approximation the decomposed fingerprint plots look similar, as seen from Fig.\u00a05b. However, relatively shorter contacts are found in form \u03b2 cf. form \u03b1, i.e. 2.62 vs 2.68\u2005\u00c5. The clear distinction between the two forms is readily noted from the decomposed fingerprint plots for the O\u22efH/H\u22efO contacts with very distinct spikes evident for form \u03b2, Fig.\u00a05c, correlating with the C\u2014H\u22efO inter\u00adactions leading to dimer formation. While beyond the sum of their respective van der Waals radii and asphericity (\u03a9) indices. All these indicators suggest that the polymorphs arise as a result of a simple inter\u00adplay between mol\u00adecular conformation and crystal-packing effects.In general, the observation of generally shorter contacts in form \u03b2 may indicate greater crystal-packing efficiency , is inter\u00admediate between those found in the polymorphic forms of (I)The most closely related structure to (I)Chemical context, biological considerations motivate ongoing investigations into the chemistry of phosphanegold(I) N-aryl-O-alkyl\u00adthio\u00adcarbamates. This notwithstanding, the relative ease of growing crystals have prompted several crystal engineering studies. Thus, correlations between Au\u22efAu (aurophilic) and solid-state luminescence responses have been made for the series of compounds, R3PAu[SC(OMe)=NC6H4NO2-p] , and bidentate phosphane analogues, Ph2P\u2013(CH2)n\u2013PPh2 for n = 1\u20134 and when the bridge is Fc (ferrocen\u00adyl) 4PPh2){AuSC(OR\u2032)=NC6H4Y-p}2] for R\u2032 = Me, Et or iPr and Y = H, NO2 or Me was undertaken =NR\u2032\u2032], for R = Ph, o-tol, m-tol or p-tol, and R\u2032\u2019 = Ph, o-tol, m-tol, p-tol or C6H4NO2-p, where it proved possible to induce a conformational change in the mol\u00adecule so that an intra\u00admolecular Au\u22ef\u03c0 inter\u00adaction formed rather than Au\u22efO species yielding binuclear mol\u00adecules also with intra\u00admolecular Au\u22ef\u03c0 inter\u00adactions in MeOH , followed by addition of the thio\u00adcarbamide, MeOC(=S)N(H)C6H4Cl3 , prepared following literature precedents : 1434 (s) \u03bd(C=N), 1180 (s) \u03bd(C\u2014O), 1098 (s) \u03bd(C\u2014S).Preparation of (I)Uiso(H) set to 1.2\u20131.5Ueq(C). The maximum and minimum residual electron density peaks of 2.04 and 1.06\u2005e\u2005\u00c5\u22123, respectively, were located 1.01 and 0.77\u2005\u00c5 from the Au atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989016010781/hb7598sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016010781/hb7598Isup2.hklStructure factors: contains datablock(s) I. DOI: 1489737CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the pyrrolidine and cyclo\u00adhexane rings exhibit envelope and chair conformations, respectively. In the crystal, C\u2014H\u22efO inter\u00adactions connect the mol\u00adecules into tape structures. 19H25NO2, the pyrrolidine ring adopts an envelope form, with the spiro C atom as the flap, while the cyclo\u00adhexane ring shows a chair form. A weak intra\u00admolecular C\u2014H\u22efO inter\u00adaction supports the mol\u00adecular conformation, generating an S(6) ring motif. In the crystal, pairs of C\u2014H\u22efO inter\u00adactions connect the mol\u00adecules into inversion dimers with an R22(16) ring motif. The dimers are linked by a second pair of C\u2014H\u22efO inter\u00adactions, enclosing an R42(12) ring motif, into a tape structure along the b axis.In the title compound, C N-hy\u00addroxy or N-alk\u00adoxy substituent have been widely explored in organic synthesis. These substances show specific and intriguing reactivity caused by a covalent bond between the electronegative heteroatoms. Among these compounds, for example, the N-alk\u00adoxy\u00adamines are known to be initiators for stable free radical polymerization = 0.1965\u2005(16)\u2005\u00c5 and \u03c6(2) = 151.8\u2005(5)\u00b0. The flap atom C5 deviates from the mean plane of other four atoms by 0.314\u2005(2)\u2005\u00c5. For the N-alk\u00adoxy-N-alkyl\u00adamide moiety, the geometry around atom N1 is a little deformed from a planar to a pyramidal configuration. The shift of atom N1 from the C2/C5/O14 plane is 0.2163\u2005(13)\u2005\u00c5, and the sum of angles for C2\u2014N1\u2014O14, O14\u2014N1\u2014C5 and C5\u2014N1\u2014C2 is 353.0\u00b0.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01Q = 0.5782\u2005(17)\u2005\u00c5, \u03b8 = 1.82\u2005(17)\u00b0, \u03c6 = 347\u2005(5)\u00b0, Q(2) = 0.0197\u2005(17)\u2005\u00c5 and Q(3) = 0.5779\u2005(17)\u2005\u00c5. The equatorially oriented C10\u2014C11 bond makes an angle of 70.60\u2005(9)\u00b0 with the normal to the Cremer & Pople plane of the cyclo\u00adhexane ring, and the vinyl group (C11=C12) is positioned in syn-periplanar geometry to the cyclo\u00adhexane framework, with a C9\u2014C10\u2014C11=C12 torsion angle of 10.9\u2005(2)\u00b0.The cyclo\u00adhexane ring (C5\u2013C10), which is spiro-fused to the pyrrolidine ring, adopts a chair form with puckering parameters of S(6) graph-set motif. No intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction is observed.An intra\u00admolecular C\u2014H\u22efO inter\u00adaction (C15\u2014H15\u22efO13) supports the mol\u00adecular conformation, generating an i; Table\u00a01b axis by weak C\u2014H\u22efO inter\u00adactions , are registered , but no compound with an N-alk\u00adoxy substituent, (c).In the Cambridge Structural Database , which is related to the title compound, (e), has also been reported , the pyrrolidine ring adopts a similar conformation to the title compound. The spiro-C atom is at the flap of the envelope, and the geometry around the N atom shows a little deformation to a pyramidal configuration with the sum of the C(carbon\u00adyl)\u2014N\u2014O, O\u2014N\u2014C and C\u2014N\u2014C(carbon\u00adyl) angles being 345.8\u2005(5)\u00b0. No intra\u00admolecular C\u2014H\u22efO inter\u00adaction is observed in (d).The structure of an et al., 2015m/z calculated for C19H25NO2Na+ [M + Na]+: 322.1783; found: 322.1779. Analysis calculated for C19H25NO2: C 76.22, H 8.42, N 4.68%; found: C 76.31, H 8.44, N 4.58%.The title compound was synthesized convergently from hex-5-en-1-ol, methyl 4-chloro-4-oxobutyrate and 1-phenyl\u00adethanol (Yamamoto Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015021209/is5431sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015021209/is5431Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015021209/is5431Isup3.cmlSupporting information file. DOI: 1435676CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two tridentate terpyridine ligands define the coordination of the Ni2+ cation, resulting in a nearly octa\u00adhedral coordination sphere, although there is not any imposed crystallographic symmetry about the Ni2+ site. The two nearly linear dicyanidoaurate(I) anions [C\u2014Au\u2014C = 179.0\u2005(2) and 178.2\u2005(2)\u00b0] contain a short aurophilic inter\u00adaction of 3.1017\u2005(3)\u2005\u00c5. The structure does not demonstrate any \u03c0\u2013\u03c0 stacking. Non-classical C\u2014H\u22efN inter\u00adactions between the cations and anions build up a three-dimensional network.The title compound, [Ni(C M(terpy)2](X) have been known since the 1970\u2032s 2][Au(CN)2]2, (I)Derivatives of the compound 2 was synthesized and analysed ]2+ cation with dicyanidoaurate(I) anions. However, the important difference between the two compounds is that there are no metal\u2013metal inter\u00adactions in the [Ni(terpy)][Au(Br)2(CN)2]2 structure containing the d8 Au(III) ion, whereas the [Ni(terpy)2][Au(CN)2]2 structure contains a d10 gold(I) dicyanidoaurate(I) anion that has a strong propensity to form aurophilic inter\u00adactions. This makes the title compound of inter\u00adest because it contains short aurophilic inter\u00adactions, contained within dimeric [Au(CN)2]2 moieties, with Au\u22efAu distances of 3.1017\u2005(3)\u2005\u00c5 \u2005\u00c5 Fig.\u00a01.D\u22efA distance of 3.5\u2005\u00c5 as the upper defined limit. Details of the inter\u00adactions can be found in Table\u00a01Structural commentary above. There are no \u03c0\u2013\u03c0 stacking inter\u00adactions in the structure.A packing diagram of the title compound is illustrated in Fig.\u00a02M Ni(NO3)2 (1\u2005ml) and 0.1 M 2,2\u2032:6\u2032.2\"-terpyridine (1\u2005ml) were mixed together. Following the mixture of these two compounds, 2\u2005ml of 0.05 M KAu(CN)2 (50:50 ethanol/water v/v) was added dropwise. A precipitate formed and the suspension was mixed thoroughly and centrifuged. The brownish-red solution was deca\u00adnted from the solid precipitate and placed in a test tube to allow for slow evaporation. After approximately one week, the formation of brownish-red crystals had begun. The grown single crystals were then gathered and isolated.Ethanol solutions of 0.1 Uiso(H) = 1.2Ueq(C) and C\u2014H distances of 0.93\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814024672/wm5063sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S1600536814024672/wm5063Isup2.hklStructure factors: contains datablock(s) I. DOI: 1033527CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit contains a quarter each of two crystallographically independent propane-1,3-di\u00adammonium dicat\u00adions, [AlF6]3\u2212 and [AlF4(H2O)2]\u2212 anions and four water mol\u00adecules. The cations, anions and three of the independent water mol\u00adecules are situated on special positions mm, while the fourth water mol\u00adecule is disordered about a mirror plane. In the crystal, inter\u00admolecular N\u2014H\u22efF and O\u2014H\u22efF hydrogen bonds link the cations and anions into a three-dimensional framework with the voids filled by water mol\u00adecules, which generate O\u2014H\u22efO hydrogen bonds and further consolidate the packing.The title compound, (C The [AlF6] octa\u00adhedron is regular whereas [AlF4(H2O)2] exhibits a pronounced distortion due to the strong influence of the crystal field created by the heteroligands (F\u2212/H2O). The value of the calculated valences (3.08 for Al1 and 3.01 for Al2) of the individual Al3+ cations 2] octa\u00adhedron is linked via N\u2014H\u22efF or O\u2014H\u22efF hydrogen bonds (Table\u00a01a axis. These chains are linked to each other by the AlF63\u2212 dications and form infinite (H2dap)[AlF4(H2O)2] layers parallel to the ac plane anions or extended 1D inorganic chains, 2D inorganic layers or 3D networks are mentioned. Eight compounds with AlF63\u2212 anions exist 2\u2212 anion 2\u2212 anions have been reported.In the Cambridge Structural Database in 40% HF (1.5\u2005ml) and ethanol (5\u2005ml). 1,3-Di\u00adamino\u00adpropane (0.54\u2005ml) was added and mild hydro\u00adthermal conditions (463\u2005K) were applied in a Teflon-lined autoclave (25\u2005ml). The resulting product was washed with ethanol and dried in air giving colourless single crystals.The title compound was prepared from a starting mixture of AlF3 and CH2 groups of the organic mol\u00adecule were fixed geometrically [N\u2014H = 0.89\u2005(1) and C\u2014H = 0.97\u2005(1)\u2005\u00c5 with Uiso(H) = 1.2Ueq]. All H atoms of the water mol\u00adecules were located from a Fourier difference map. The O\u2014H distances and H\u2014O\u2014H angles were fixed [O\u2014H = 0.84\u2005(1) and H\u22efH = 1.34\u2005(1)\u2005\u00c5 with Uiso(H) =1.5Ueq(O)]. The water mol\u00adecule OW5 is disordered over two positions with the occupanies fixed to 0.5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814024155/cv5471sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S1600536814024155/cv5471Isup2.hklStructure factors: contains datablock(s) I. DOI: 1032262CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atom in the title complex is surrounded by a distorted Cl2N4 coordination set. In the crystal structure, adjacent complex mol\u00adecules are connected through C\u2014H\u22efCl hydrogen-bonding inter\u00adactions into a layered arrangement parallel to (100). Additional C\u2014H\u22efBr hydrogen-bonding inter\u00adactions along with \u03c0\u2013\u03c0 stacking inter\u00adactions complete a three-dimensional supra\u00admolecular network.The Ru 2(C12H9BrN2)2] or [RuCl2(PM-BrA)2] (PM-BrA = 4-bromo-N-(2\u2032-pyridyl\u00admethyl\u00adene)aniline), the RuII cation is located on a centre of inversion and is surrounded by four N atoms of two PM-BrA ligands in the equatorial plane and by two Cl atoms in a trans axial arrangement, displaying a distorted octa\u00adhedral coordination environment. Two C atoms in the benzene ring of the PM-BrA ligand are equally disordered over two sets of sites. The benzene and pyridine rings of the PM-BrA ligand are oriented at dihedral angles of 62.1\u2005(10) and 73.7\u2005(11)\u00b0 under consideration of the two orientations of the disordered benzene ring. In the crystal, the complex mol\u00adecules are connected via C\u2014H\u22efCl hydrogen-bonding inter\u00adactions into a layered arrangement parallel (100). C\u2014H\u22efBr hydrogen bonding and weak aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions complete a three-dimensional supra\u00admolecular network.In the title complex, [RuCl Their complexes have found utility in a wide range of applications \u2005\u00c5 with an N1\u2014Ru1\u2014N2 bite angle of 76.9\u2005(1)\u00b0. The reduced bite angle of the chelating ligand is one of the main factors accounting for the distortion from the ideal octa\u00adhedral geometry of the coordination polyhedron, with the the largest cis angle being 103.1\u2005(2)\u00b0. The Ru\u2014N bond lengths are 2.073\u2005(5) and 2.084\u2005(5)\u2005\u00c5, and the Ru\u2014Cl bond length is 2.3908\u2005(14)\u2005\u00c5, in agreement with those observed in the structures of similar compounds re Fig.\u00a01. The cooX inter\u00adactions are compiled in Table\u00a01In the crystal, weak inter\u00admolecular C\u2014H\u22efCl hydrogen-bonding inter\u00adactions between the C atoms of the benzene ring and the Cl atoms connect the complex mol\u00adecules into a supra\u00admolecular layered arrangement parallel to (100) Fig.\u00a02. As showtrans-[RuCl2(Hpyrimol)2] (Hpyrimol = 4-methyl-2-N-(2-pyridyl\u00admethyl\u00adene)amino\u00adphenol) with a closely related Schiff base N2 donor set for each ligand has been reported aniline in dry methanol (5\u2005ml) was placed in a test tube. A solution of RuCl3 in dry methanol (5\u2005ml) was then carefully layered on the top of a methano\u00adlic solution. After slow diffusion at room temperature for three days, pale-green plate- or block-like crystals of complex (I)A solution of the ligand 4-bromo-Uiso(H) = 1.2Ueq(C) using a riding model with C\u2014H = 0.95\u2005\u00c5. C atoms C11 and C12 and attached H atoms in the benzene ring are disordered over two set of sites and were refined using a split model with equal occupancy.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901501556X/wm5204sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901501556X/wm5204Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901501556X/wm5204Isup3.cdxSupporting information file. DOI: 1419653CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E conformation with respect to the azomethine C=N bond and the aromatic rings are inclined to one another by 3.29\u2005(4)\u00b0. In the crystal, mol\u00adecules are linked via C\u2014H\u22efO hydrogen bonds, forming zigzag chains along [10-1].The title compound has an 14H13NO2 {systematic name: (E)-2-[(4-meth\u00adoxy\u00adbenzyl\u00adidene)amino]\u00adphenol}, is a product of the condensation reaction between 4-meth\u00adoxy\u00adbenzaldehyde and 2-amino\u00adphenol. The mol\u00adecule adopts an E conformation with respect to the azomethine C=N bond and is almost planar, the dihedral angle between the two substituted benzene rings being 3.29\u2005(4)\u00b0. The meth\u00adoxy group is coplanar with the benzene ring to which it is attached, the Cmeth\u00adyl\u2014O\u2014C\u2014C torsion angle being \u22121.14\u2005(12)\u00b0. There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond generating an S(5) ring motif. In the crystal, mol\u00adecules are linked via C\u2014H\u22efO hydrogen bonds, forming zigzag chains along [10-1]. The chains are linked via C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional structure.The title aza\u00adstilbene derivative, C The mol\u00adecule is almost planar with a dihedral angle of 3.29\u2005(4)\u00b0 between the two substituted benzene rings. The meth\u00adoxy group is co-planar with the benzene ring to which it is attached, the C14\u2014O1\u2014C4\u2014C5 torsion angle being \u22121.14\u2005(12)\u00b0. There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond ring motif. The bond lengths are comparable with those found for some closely related structures . Colourless block-shaped crystals, suitable for X-ray structure analysis, were obtained by recrystallization from methanol by slow evaporation at room temperature after several days (m.p. 388\u2013390\u2005K).A solution of 4-meth\u00adoxy\u00adbenzaldehyde in water (20\u2005ml) and 2-amino\u00adphenol in water (20\u2005ml) were mixed and stirred at room temperature for around 8\u2005h until a white precipitate appeared. The resulting white solid was filtered, washed several times with cold ethanol and then dried 3OH) \u03bbmax (log\u220a): 275 (1.93), 340 (0.61)\u2005nm; FT\u2013IR (KBr) \u03bd: 3337, 1595, 1510, 1248, 1027\u2005cm\u22121.; 1H NMR \u03b4, p.p.m.: 8.87 , 8.61 , 7.98 , 7.18 , 7.06 , 7.03 , 6.83 , 6.09 , 3.84 . The UV\u2013Vis spectroscopic data showed absorption bands of an aza\u00adstilbene (275 and 340\u2005nm) while the FT\u2013IR spectrum exhibited the stretching vibrations of O\u2014H (3337\u2005cm\u22121), C=N (1595\u2005cm\u22121), C=C (1510\u2005cm\u22121), C\u2014N (1248\u2005cm\u22121) and C\u2014O (1027\u2005cm\u22121). The successful synthesis was also supported by the 1H NMR spectroscopic data, which showed the characteristic signals of an olefinic proton at 8.61 and para-substituted aromatic protons at 7.98 and 7.06 , respectively. Moreover the 1H NMR spectrum also showed typical signals of ortho-substituted aromatic protons at 7.18 , 7.03 , 6.83 and 6.09 and a meth\u00adoxy proton at 3.84 .UV\u2013Vis = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015008348/su5124sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015008348/su5124Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015008348/su5124Isup3.cmlSupporting information file. DOI: 1062128CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I atom within an S(di\u00adthio\u00adcarbamate) and P(phosphane) donor set.The centrosymmetric mol\u00adecule features a linearly coordinated Au 34H28FeP2)[Au(C5H8NS2)]2}, comprises half a mol\u00adecule, with the full mol\u00adecule being generated by the application of a centre of inversion. The independent AuI atom is coordinated by thiol\u00adate S and phosphane P atoms that define an approximate linear geometry [S\u2014Au\u2014P = 169.35\u2005(3)\u00b0]. The deviation from the ideal linear is traced to the close approach of the (intra\u00admolecular) non-coordinating thione S atom [Au\u22efS = 3.1538\u2005(8)\u2005\u00c5]. Supra\u00admolecular layers parallel to (100) feature in the crystal packing, being sustained by phen\u00adyl\u2013thione C\u2014H\u22efS inter\u00adactions, with the non-coordinating thione S atom in the role of a dual acceptor. Layers stack with no specific inter\u00adactions between them.The asymmetric unit of the title compound, {(C R3PAu(S2CNR\u20322), date back over a decade CH2CH2OH], R = Ph and Cy, exhibited specific activity against Gram-positive bacteria while the R = Et derivative displayed broad-range activity against both Gram-positive and Gram-negative bacteria. Motivated by observations that 1,1\u2032-bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)ferrocene (dppf) derivatives also possess biological activity 4]}2, (I)Investigations into the potential anti-cancer activity of phosphanegold(I) di\u00adthio\u00adcarbamates, II atom in dppf{Au[S2CN(CH2)4]}2, (I)I central atom exists in the anti\u00adcipated linear geometry defined by thiol\u00adate-S and phosphane-P atoms. The Au\u2014S1 bond length is considerably longer than the Au\u2014P1 bond, i.e. 2.3378\u2005(8) cf. 2.2580\u2005(8)\u2005\u00c5. The di\u00adthio\u00adcarbamate ligand is orientated to place the S2 atom in close proximity to the AuI atom. However, the resulting intra\u00admolecular Au\u22efS2 inter\u00adaction is long at 3.1538\u2005(8)\u2005\u00c5, consistent with a monodentate mode of coordination for the di\u00adthio\u00adcarbamate ligand. The pattern of C1\u2014S1, S2 bond lengths supports this conclusion in that the strongly bound S1 atom forms a longer, i.e. weaker, C1\u2014S1 bond [1.757\u2005(3)\u2005\u00c5] cf. with C1\u2014S2 of 1.689\u2005(3)\u2005\u00c5. Nevertheless, the close approach of the S2 atom to the AuI central atom is correlated with the deviation from the ideal linear geometry, i.e. S1\u2014Au\u2014P1 is 169.35\u2005(3)\u00b0. The FeDatabase survey. The pyrrolidine ring is twisted about the C2\u2014C3 bond. Owing to being located on a centre of inversion, the FeII atom is equidistant from the ring centroids of the Cp rings and the Cg\u2014Fe\u2014Cgi angle is constrained by symmetry to be 180\u00b0; symmetry operation (i): 1\u00a0\u2212\u00a0x, \u2212y, 2\u00a0\u2212\u00a0z. Again, from symmetry, the Cp rings have a staggered relationship.Similar features are noted in related structures as outlined below in the bc plane, Fig.\u00a02a axis, i.e. 10.9635\u2005(4)\u2005\u00c5, and the layers stack along this axis with no directional inter\u00adactions between them, Fig.\u00a03In the crystal packing, the most prominent inter\u00adactions are of the type C\u2014H\u22efS. Data for the phenyl-C\u2014H\u22efS(thione) inter\u00adactions are collected in Table\u00a01i.e. Ph3PAu(S2CNEt2), by Wijnhoven et al. (19723P 2\u00b72CHCl3, bis\u00ad[chlorido\u00adgold(I)] (synthesized by the reduction of KAuClUiso(H) set to 1.2Ueq(C). The maximum and minimum residual electron density peaks of 1.57 and 1.11\u2005e\u2005\u00c5\u22123, respectively, were located 0.92 and 0.79\u2005\u00c5 from the Au atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015016382/vn2097sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989015016382/vn2097Isup2.hklStructure factors: contains datablock(s) I. DOI: 1421954CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Sodium bicarbonate intake has been shown to improve exercise tolerance, but the effects on high-intensity intermittent exercise are less clear. Thus, the aim of the present study was to determine the effect of sodium bicarbonate intake on Yo-Yo intermittent recovery test level 2 performance in trained young men.2max: 61.3\u2009\u00b1\u20093.3 mlO2\u2009\u00b7\u2009kg\u22121\u2009\u00b7\u2009min\u22121; means\u2009\u00b1\u2009SEM) performed the Yo-Yo intermittent recovery test level 2 (Yo-Yo IR2) on two separate occasions in randomized order with (SBC) and without (CON) prior intake of sodium bicarbonate (0.4 g\u2009\u00b7\u2009kg\u22121 body weight). Heart rate and rating of perceived exertion (RPE) were measured during the test and venous blood samples were taken frequently.Thirteen men aged 23\u2009\u00b1\u20091 year . Blood lactate was 0.9\u2009\u00b1\u20090.1 and 0.8\u2009\u00b1\u20090.1 mmol\u2009\u00b7\u2009l\u22121 at baseline and increased to 11.3\u2009\u00b1\u20091.4 and 9.4\u2009\u00b1\u20090.8 mmol\u2009\u00b7\u2009l\u22121 at exhaustion in SBC and CON, respectively, being higher (P\u2009=\u20090.03) in SBC. Additionally, peak blood lactate was higher (P\u2009=\u20090.02) in SBC than in CON (11.7\u2009\u00b1\u20091.2 vs 10.2\u2009\u00b1\u20090.7 mmol\u2009\u00b7\u2009l\u22121). Blood glucose, plasma K+ and Na+ were not different between trials. Peak heart rate reached at exhaustion was 197\u2009\u00b1\u20093 and 195\u2009\u00b1\u20093 bpm in SBC and CON, respectively, with no difference between conditions. RPE was 7\u00a0% lower (P\u2009=\u20090.003) in SBC than in CON after 440 m, but similar at exhaustion (19.3\u2009\u00b1\u20090.2 and 19.5\u2009\u00b1\u20090.2).Yo-Yo IR2 performance was 14\u00a0% higher (P\u2009=\u20090.04) in SBC than in CON . Blood pH and bicarbonate were similar between trials at baseline, but higher (P\u2009=\u20090.003) immediately prior to the Yo-Yo IR2 test in SBC than in CON (7.44\u2009\u00b1\u20090.01 vs 7.32\u2009\u00b1\u20090.01 and 33.7\u2009\u00b1\u20093.2 vs 27.3\u2009\u00b1\u20090.6 mmol\u2009\u00b7\u2009lIn conclusion, high-intensity intermittent exercise performance is improved by prior intake of sodium bicarbonate in trained young men, with concomitant elevations in blood alkalosis and peak blood lactate levels, as well as lowered rating of perceived exertion. The remaining whole blood was then centrifuged at 4000 rpm for 3 min before plasma was extracted and stored on ice for ~30 min prior to being frozen at -80\u2009Denmark) .RPE was assessed at 160, 280, 440 m and exhaustion according to the 20-stage Borg scale .P\u2009<\u20090.05. Results are presented as mean\u2009\u00b1\u2009SEM unless stated otherwise.Differences between SBC and CON in Yo-Yo IR2 performance were analyzed using a paired-samples t-test. Differences between SBC and CON in blood and plasma variables were analyzed using a two-way ANOVA with repeated measures (supplement x time). If a significant F-value was observed, a Tukey post-hoc test was used to identify the points of difference. Statistical significance was accepted at Yo-Yo IR2 performance was 735\u2009\u00b1\u200961 m in SBC, which was 14\u00a0% higher (P\u2009=\u20090.04) than in CON to 7.44\u2009\u00b1\u20090.01 in SBC before the Yo-Yo IR2 test and decreased (P\u2009=\u20090.005) in CON in SBC before the Yo-Yo IR2 test (33.7\u2009\u00b1\u20093.2 mmol\u2009\u00b7\u2009l\u22121) and was unchanged in CON (27.3\u2009\u00b1\u20090.6 mmol\u2009\u00b7\u2009l\u22121) compared to baseline. At exhaustion, blood [bicarbonate] declined (P\u2009<\u20090.001) in SBC and CON (25.2\u2009\u00b1\u20092.6 and 20.2\u2009\u00b1\u20090.8 mmol\u2009\u00b7\u2009l\u22121). Pre-exercise and exhaustive blood [bicarbonate] was higher (P\u2009=\u20090.01 and 0.03) in SBC than in CON 11.3\u2009\u00b1\u20091.4 and 9.4\u2009\u00b1\u20090.8 mmol\u2009\u00b7\u2009l\u22121 at exhaustion in SBC and CON, respectively, being higher (P\u2009=\u20090.03) in SBC (Table\u00a0\u22121).Blood lactate was 0.9\u2009\u00b1\u20090.1 and 0.8\u2009\u00b1\u20090.1 mmol\u2009\u00b7\u2009l\u22121 at baseline to 5.1\u2009\u00b1\u20090.5 and 5.2\u2009\u00b1\u20090.6 mmol\u2009\u00b7\u2009l\u22121 at exhaustion, with no difference between trials (Table\u00a0Blood glucose rose (P\u2009<\u20090.05) from 4.2\u2009\u00b1\u20090.4 and 4.1\u2009\u00b1\u20090.2 mmol\u2009\u00b7\u2009l+] rose (P\u2009<\u20090.05) from 4.3\u2009\u00b1\u20090.4 and 4.2\u2009\u00b1\u20090.4 mmol\u2009\u00b7\u2009l\u22121 at baseline to 5.7\u2009\u00b1\u20090.5 and 5.8\u2009\u00b1\u20090.6 mmol\u2009\u00b7\u2009l\u22121 at exhaustion in SBC and CON, respectively, with no differences between trials during the test, reaching 197\u2009\u00b1\u20093 and 195\u2009\u00b1\u20093 bpm at exhaustion in SBC and CON, respectively. No difference in heart rate response between conditions was determined during or after the test.RPE was not different between trials after 160 and 280 m , but was 7\u00a0% lower (P\u2009=\u20090.003) after 440 m in SBC than in CON (16.8\u2009\u00b1\u20090.4 vs 17.9\u2009\u00b1\u20090.3). At exhaustion, RPE was similar in SBC and CON (19.3\u2009\u00b1\u20090.2 and 19.5\u2009\u00b1\u20090.2).+ and Na+ as well as cardiovascular loading during high-intensity intermittent exercise were unaffected by sodium bicarbonate intake.In the present study, we observed that prior intake of sodium bicarbonate in capsular form using a protocol with gradual intake enhanced high-intensity intermittent exercise performance in young trained males. The performance improvement after sodium bicarbonate ingestion was accompanied by an elevated blood alkalosis and concentration of bicarbonate. In addition, blood lactate concentrations at exhaustion and peak values reached during the experimental protocol were higher, while the rating of perceived exertion was lower during intense exercise after sodium bicarbonate supplementation. In contrast, blood glucose, plasma KPerformance in the Yo-Yo IR2 test increased by 14\u00a0% after sodium bicarbonate intake, which is comparable to a 16\u00a0% increase after caffeine intake in a comparable athlete population , but lowIn the present study, we did not apply a placebo, since the gastrointestinal effects of high doses of bicarbonate are usually easy to trace . A poten+ gradient and [Na+] between the sodium bicarbonate and control trials. However, since the potential fatiguing effect of a homeostatic imbalance in these ions is exerted in the muscle interstitial compared to intracellular environment [+ regulation, Street et al. [+ accumulation rate during intense exercise was attenuated after drug-induced alkalosis.Fatigue development during high-intensity intermittent exercise may be caused by a complex interplay between intra- and extracellular concentrations and gradients of ions such as K\u2212 and H+ , 29. In ironment \u201332, a soironment . In suppt et al. found, umax and blood lactate and plasma K+ were markedly elevated. Moreover, although further distance was covered before exhaustion, the RPE scores remained the same as in the control trial at the point of fatigue. Thus, a higher performance level was achieved while reporting an equal level of perceived fatigue at exhaustion. Peripheral alterations are likely to lead to modulation of neural strategies, for example via group III and IV muscle afferents, widely distributed through muscle and responsive to a variety of chemical stimuli, including altered H+ [In the present study, RPE was lowered after 440 m of running in the Yo-Yo IR2 test in the sodium bicarbonate trial compared to the control, despite the fact that the heart rate and blood lactate concentration at this time-point were similar between trials. This may suggest that centrally mediated mechanisms were affected. The participants experienced less exertion late in the Yo-Yo IR2 test in the SBC trial when the heart rate was >95\u00a0% of HRtered H+ , 34, whitered H+ , 35.\u22121 body weight) improved high-intensity intermittent exercise performance in trained young men, with concomitant increased blood alkalosis. Sodium bicarbonate supplementation increased blood lactate levels at exhaustion and lowered rating of perceived exertion during intense intermittent exercise. The results indicate a link between improved fatigue resistance during high-intensity intermittent exercise and a sodium-bicarbonate-induced improved buffer capacity that may affect perceived exertion during intense intermittent exhaustive exercise.High-dose sodium bicarbonate intake (0.4 g\u2009\u00b7\u2009kg"} +{"text": "Single crystals were obtained from HSO3F by slow cooling in a sealed tube. The mol\u00adecular structure is characterized by the Xe atom covalently bonded to two O atoms of two fluoro\u00adsulfate tetra\u00adhedra in an almost linear fashion [O\u2014Xe\u2014O = 179.13\u2005(4)\u00b0]. The crystal packing is strongly influenced by inter\u00admolecular van der Waals forces.Thermally unstable Xe(SO For XeF(OSO2F), partial ionic bonding (XeF+\u00b7OSO2F\u2212) was discussed. Obviously, both XeF2 and Xe(SO3F)2 have a higher covalent character. The S\u2014O bonds in Xe(SO3F)2 involving the O atoms that are also bonded to the xenon atom (S1\u2014O1 and S2\u2014O4) are about 0.1\u2005\u00c5 longer than the terminal S\u2014O bonds has fewer contacts was warmed to 273\u2005K and the PFA tube placed in a dewar filled with 273\u2005K ethanol and cooled slowly to 193\u2005K in a freezer. The light-yellow single crystals of Xe(SO3F)2 that had formed were deca\u00adnted off and mounted in a cold nitro\u00adgen stream. At 100\u2005K, the crystals are colorless. The compound decomposes rapidly in moist air and can ignite organic materials.550\u2005mg fluoro\u00adsulfuric acid were placed in a 8\u2005mm PFA tube. 170\u2005mg (1\u2005mmol) of XeFCrystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015004788/wm5134sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015004788/wm5134Isup2.hklStructure factors: contains datablock(s) I. DOI: 1052852CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit is composed of a copper(II) tetra\u00adacetate paddle-wheel complex, a Cl\u2212 anion situated on a twofold rotation axis, half a 1,3-bis\u00adimidazolium cation (the whole mol\u00adecule being generated by twofold rotation symmetry) and one and a half of a di\u00adchloro\u00admethane solvent mol\u00adecule (one being located about a twofold rotation axis). The central metal-organic framework comprises of a tetra\u00adnuclear copper(II) acetate \u2018paddle-wheel\u2019 complex which arises from the dimerization of the copper(II) tetra\u00adacetate core comprising of three \u03bc2-bidentate acetate and one \u03bc3-tridentate acetate ligands per binuclear paddle-wheel complex. Both CuII atoms of the binuclear component adopt a distorted square-pyramidal coordination geometry (\u03c4 = 0.04), with a Cu\u22efCu separation of 2.6016\u2005(2)\u2005\u00c5. The apical coordination site of one CuII atom is occupied by an O atom of a neighbouring acetate bridge [Cu\u2014O = 2.200\u2005(2)\u2005\u00c5], while that of the second CuII atom is occupied by a bridging chloride ligand [Cu\u22efCl = 2.4364\u2005(4)\u2005\u00c5]. The chloride bridge is slightly bent with respect to the Cu\u22efCu inter\u00adnuclear axis [Cu\u2014Cl\u2014Cu = 167.06\u2005(6)\u00b0] and the tetra\u00adnuclear units are located about a twofold rotation axis, forming the one-dimensional polymer that propagates along [101]. Charge neutrality is maintained by the inclusion of the 1,3-bis\u00adimidazolium cation within the crystal lattice. In the crystal, the cation and di\u00adchloro\u00admethane solvent mol\u00adecules are linked to the coordin\u00adation polymer by various C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds. There are no other significant inter\u00admolecular inter\u00adactions present.The title copper(II) complex, {(C For the al. 2005; Bull et al. 2008. For det al. 1964; Zhang e al. 2005; Cotton al. 2000, and for al. 2003. For exa al. 1973; Ackerma al. 2000. For chl al. 2015. For imi al. 2015. For the al. 1984.27H37N2)[Cu4(C2H3O2)8Cl]\u00b73CH2Cl2 \u00c5b = 13.146 (2) \u00c5c = 23.607 (3) \u00c5\u03b2 = 117.122 (4)\u00b0V = 6103.5 (13) \u00c53Z = 4K\u03b1 radiationMo \u22121\u03bc = 1.74 mmT = 100 K0.22 \u00d7 0.13 \u00d7 0.05 mmBruker SMART CCD area-detector diffractometerSADABS; Bruker, 2001Tmin = 0.700, Tmax = 0.918Absorption correction: multi-scan (26111 measured reflections7274 independent reflectionsI > 2\u03c3(I)5215 reflections with Rint = 0.079R[F2 > 2\u03c3(F2)] = 0.054wR(F2) = 0.098S = 0.987274 reflections348 parametersH-atom parameters constrainedmax = 0.70 e \u00c5\u22123\u0394\u03c1min = \u22120.46 e \u00c5\u22123\u0394\u03c1SMART used to solve structure: SHELXS97 I, global. DOI: 10.1107/S2056989015013675/su5152Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015013675/su5152fig1.tif. DOI: A view of the mol\u00adecular structure of the asymmetric unit of the title compound. Displacement ellipsoids are drawn at the 50% probability level.Click here for additional data file.10.1107/S2056989015013675/su5152fig2.tifx y z x y z x y z x y z x y z . DOI: x, y, \u2212z\u00a0+\u00a0x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a01; (c) x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a0x, y, \u2212z\u00a0+\u00a0x\u00a0+\u00a01, y, \u2212z\u00a0+\u00a0A view of the tetra\u00adnuclear paddle-wheel unit of the title polymeric compound [symmetry codes: (a) \u2212Click here for additional data file.10.1107/S2056989015013675/su5152fig3.tifb 2 2 . DOI: b axis of the crystal packing of title compound. Colour code: coordination polymer black, organic cation red; CH2Cl2 solvent mol\u00adecules green and blue.A view along the 999046CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information: interactive version of Fig. 1dEnhanced figure:"} +{"text": "The O\u2014O bond length of the peroxide is 1.485\u2005(6)\u2005\u00c5 and the mid-point of this bond is located at the inversion centre of the dimer. The U atom exhibits a distorted hexa\u00adgonal\u2013bipyramidal coordination geometry with two uran\u00adyl(VI) O atoms occupying the axial positions and one O atom of the monodentate nitrate ion, both O atoms of the peroxide ion and the three N atoms of the chelating tridentate 2,2\u2032:6\u2032,2\u2032\u2032-terpyridine (terpy) ligand in the equatorial positions. Two of the N atoms of the terpy ligand lie above and below the mean plane containing the equatorial ligand atoms and the U atom . The dihedral angle between the terpy ligand and the mean plane is 35.61\u2005(7)\u00b0. The bond lengths around the U atom decrease in the order U\u2014N > U\u2014Onitrate > U\u2014Operoxo > U=O. The dimeric complexes pack in a three-dimensional network held together by weak \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.659\u2005(3)\u2005\u00c5] between pyridyl rings of the terpy ligands in neighbouring dimers, together with inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. Weak intra\u00admolecular C\u2014H\u22efO inter\u00adactions are also observed.In the title dimeric complex, [{UO For the al. 2001; Goff et al. 2008; John et al. 2004; Sigmon al. 2009; Takao & al. 2009. For theuwer 2004.2(NO3)2(O2)O4(C15H11N3)2] = 0.030wR(F2) = 0.058S = 0.894695 reflections235 parametersH-atom parameters constrainedmax = 2.24 e \u00c5\u22123\u0394\u03c1min = \u22121.57 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXS97 I, global. DOI: 10.1107/S2056989015007987/cq2015Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015007987/cq2015fig1.tif2 3 15 11 3 2 2 x y z . DOI: 2(NO3)(C15H11N3)}2O2]. Displacement ellipsoids are drawn at the 50% probability level. H atoms are omitted for clarity. Structure of the dimer [{UOClick here for additional data file.10.1107/S2056989015007987/cq2015fig2.tif2 3 15 11 3 2 2 . DOI: 2(NO3)(C15H11N3)}2O2]. Dashed lines and dotted lines are \u03c0\u2013\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions, respectively.Packing diagram of [{UO1061056CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "These complexes are relevant to catalysis for CO2 reduction.The structures of two facially coordinated Group VII metal complexes, fac-[ReCl(C10H8N2O2)(CO)3]\u00b7C4H8O (I\u00b7THF) and fac-[MnBr(C10H8N2O2)(CO)3]\u00b7C4H8O (II\u00b7THF), are reported. In both complexes, the metal ion is coordinated by three carbonyl ligands, a halide ligand, and a 6,6\u2032-dihy\u00addroxy-2,2\u2032-bi\u00adpyridine ligand in a distorted octa\u00adhedral geometry. Both complexes co-crystallize with a non-coordinating tetra\u00adhydro\u00adfuran (THF) solvent mol\u00adecule and exhibit inter\u00admolecular but not intra\u00admolecular hydrogen bonding. In both crystal structures, chains of complexes are formed due to inter\u00admolecular hydrogen bonding between a hy\u00addroxy group from the 6,6\u2032-dihy\u00addroxy-2,2\u2032-bi\u00adpyridine ligand and the halide ligand from a neighboring complex. The THF mol\u00adecule is hydrogen bonded to the remaining hy\u00addroxy group.The structures of two facially coordinated Group VII metal complexes, In this paper we report the synthesis and structural characterization of fac-[Re(CO)3Cl] as well as the synthesis and structural characterization of the related and previously unknown complex, fac-[Mn(CO)3Br]. Both complexes co-crystallize with a tetrahydrofuran (THF) solvent molecule.The fac-[Re(CO)3Cl]\u00b7THF (I\u00b7THF) and [Mn(CO)3Br]\u00b7THF (II\u00b7THF), respectively. Complexes I and II exhibit distorted octa\u00adhedral geometries and contain primary coord\u00adination spheres similar to those of other fac-[Re(\u03b1-di\u00adimine)(CO)3Cl] and fac-[Mn(\u03b1-di\u00adimine)(CO)3Br] complexes, including [Re(bi\u00adpyridine)(CO)3Cl] (III) and 18.93\u2005(3)\u00b0 for the Re and Mn complexes, respectively. Neither I\u00b7THF nor II\u00b7THF exhibit intra\u00admolecular hydrogen bonding.In I, the bi\u00adpyridine rings present a bite angle of 74.09\u2005(8)\u00b0 to Re, similar to that found in III [74.41\u2005(9)\u00b0] and IV [74.9\u2005(2)\u00b0]. The bi\u00adpyridine\u2013Mn bite angle in II, 78.35\u2005(4)\u00b0, is similar to that in V [79.0\u2005(5)\u00b0]. The bi\u00adpyridine ligands are not strictly planar. The dihedral angles between the pyridine rings are 11.68\u2005(9)\u00b0 in I and 9.49\u2005(5)\u00b0 in II. Additionally, the bi\u00adpyridine ligands are not oriented strictly perpendicularly to the coordination planes of the metal ions. The dihedral angles between the mean plane through the \u03b1-di\u00adimine ligands and the COa axis is formed by an O\u2014H\u22efCl hydrogen bond between a hy\u00addroxy group (O16\u2014H16) and the chloride ligand from the neighboring complex. The other hy\u00addroxy group (O26\u2014H26) is hydrogen-bonded to the O atom of the THF mol\u00adecule. The nearest pyridine rings between neighboring complexes have centroid\u2013centroid distances of 3.9448\u2005(16)\u2005\u00c5, longer than the maximum distance typically given for \u03c0\u2013\u03c0 inter\u00adactions is hydrogen-bonded to O1S of the solvent THF mol\u00adecule. There are weak \u03c0\u2013\u03c0 stacking inter\u00adactions between pairs of complexes from neighboring chains. The centroid\u2013centroid distance between pairs of pyridine rings is 3.7019\u2005(9)\u2005\u00c5 and the angle between the ring normal and the vector between the ring centroids is 9.3\u00b0, within the parameters typically given for such \u03c0\u2013\u03c0 inter\u00adactions 5 and ReCl(CO)5 were purchased commercially and used as received. The ligand 6,6\u2032-dihy\u00addroxy-2,2\u2032-bi\u00adpyridine was synthesized according to the synthetic procedure of Umemoto et al. were heated at 333\u2005K in 50\u2005mL methanol under nitro\u00adgen for five\u2005h. The flask was covered with aluminum foil to keep out light. The reaction was then allowed to cool to room temperature and the solvent was removed under vacuum to give a yellow precipitate. Slow cooling of a hot THF solution of the complex in a glove box under a nitro\u00adgen atmosphere gave yellow plate-shaped crystals suitable for single crystal X-ray diffraction. Due to limited solubility of the complex in THF, this method could not be used for a bulk recrystallization of the complex.I\u00b7THF: 6,6\u2032-dihy\u00addroxy-2,2\u2032-bi\u00adpyridine and ReCl(CO)5 were heated at 333\u2005K in 24\u2005mL methanol under nitro\u00adgen for five\u2005h. The flask was covered with aluminum foil to keep out light. The reaction was then allowed to cool to room temperature and the solvent was removed under vacuum to give an orange precipitate. The complex was recrystallized in bulk by layering pentane on a THF solution of the complex in a glove box under a nitro\u00adgen atmosphere at room temperature, giving the pure product in near qu\u00adanti\u00adtative yield. Slow diffusion of diethyl ether into a THF solution of the complex in a glove box under a nitro\u00adgen atmosphere gave yellow rod-shaped crystals suitable for single crystal X-ray diffraction.II\u00b7THF: 6,6\u2032-dihy\u00addroxy-2,2\u2032-bi\u00adpyridine and MnBr(CO)Uiso(H) = 1.5 Ueq(O). C-bound H atoms were placed in calculated positions and refined with riding coordinates, with Uiso(H) = 1.2 Ueq(C). In I\u00b7THF, disorder occurs for one carbon and six hydrogens of the THF solvent with occupancies of 0.748\u2005(11) and 0.252\u2005(11). Rigid bond (DELU) and similar ADP (SIMU) restraints were used for atoms O1S, C1S, C2S, C3T, C3S and C4S.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016011841/is5457sup1.cifCrystal structure: contains datablock(s) Re_complex, Mn_complex, global. DOI: 10.1107/S2056989016011841/is5457Re_complexsup4.hklStructure factors: contains datablock(s) Re_complex. DOI: 10.1107/S2056989016011841/is5457Mn_complexsup5.hklStructure factors: contains datablock(s) Mn_complex. DOI: 1495018, 1495017CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The largest deviation from the ideal octa\u00adhedral geometry is reflected by the small N\u2014Fe\u2014N bite angle of 76.0\u2005(1)\u00b0. The Fe\u2014N coordination bonds have markedly different lengths [2.1361\u2005(17) and 2.243\u2005(2)\u2005\u00c5], with the shorter one to the pyrimidine N atom. The four Fe\u2014O coordination bond lengths vary from 2.1191\u2005(18) to 2.1340\u2005(17)\u2005\u00c5. In the crystal, the cations and anions are arranged by means of medium-strength O\u2014H\u22efO hydrogen bonds into layers parallel to the ab plane. Neighbouring layers further inter\u00adconnect by N\u2014H\u22efO hydrogen bonds involving the imidazole fragment as donor group to one sulfate O atom as an acceptor. The resulting three-dimensional network is consolidated by C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions.In the title compound, [Fe(C The distorted octa\u00adhedral environment of the central FeII ion is defined by two N donor atoms of the pyim ligand and by the O atoms of two water mol\u00adecules in the equatorial plane, while the two remaining water mol\u00adecules coordinate at the axial sites. The bite angle N1\u2014Fe\u2014N2 of 76.04\u2005(7)\u00b0 shows the most significant deviation from the ideal octa\u00adhedral geometry, with the other coordination angles deviating by 0.21\u2005(7) to 11.91\u2005(7)\u00b0.Fig.\u00a012O)4]SO4 complex where dmbpy is 5,5\u2032-dimethyl-2,2\u2032-bi\u00adpyridine \u00b0. The dihedral angle of 5.5\u2005(1) \u00b0 between the aromatic rings of the pyim ligand is within the range of the values reported for the eight independent mol\u00adecules in the crystal structure of the non-coordinating ligand SO4 complex, which range from 2.079\u2005(2) to 2.110\u2005(2)\u2005\u00c5.The Fe\u2014N coordination bonds with the chelate ligand have markedly different lengths, Fe\u2014N1 = 2.243\u2005(2) and Fe\u2014N2 = 2.1361\u2005(17)\u2005\u00c5, which are also dissimilar to those in the previously reported [Fe(dmbpy)(H2O)4]2+ unit is surrounded by five [SO4]2\u2212 anions. Similarly to the crystal structure of [Fe(dmbpy)(H2O)4]SO4, pairs of axially and equatorially coordinating water mol\u00adecules bind to pairs of O acceptor atoms from the same [SO4]2\u2212 group, forming eight medium-strength inter\u00adactions = \u2212x\u00a0+\u00a0y\u00a0+\u00a0z; Cg1 is the centroid of the imidazole ring] and weak \u03c0\u2013\u03c0 inter\u00adactions . C\u2014H\u22efO inter\u00adactions are also observed s Table\u00a01. These hne Fig.\u00a02. Additione Fig.\u00a02. The vicd Table\u00a01.H-imidazole and potassium tri\u00adcyano\u00admethanide KC(CN)3 in water-ethanol . The mixture was transferred to a Teflon-lined autoclave and heated at 423\u2005K for 48\u2005h. The autoclave was then allowed to cool to ambient temperature. Block-like yellow crystals of (I)The title compound was obtained under hydro\u00adthermal conditions from a mixture of iron(II) sulfate hepta\u00adhydrate , 2-(pyridin-2-yl)-1Uiso(H) = 1.2Ueq(C). The H atoms attached to O and N atoms were located in a difference Fourier map and were refined isotropically.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015004417/wm5132sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015004417/wm5132Isup2.hklStructure factors: contains datablock(s) I. DOI: 1051905CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the compound, a succession of MnII ions (situated on inversion centers) adopting a distorted octa\u00adhedral coordination and bridged by oxalate ligands forms parallel zigzag chains running along the c axis. These chains are inter\u00adconnected through O\u2014H\u22efO hydrogen-bonding inter\u00adactions to form anionic layers parallel to (010). Individual layers are held together via strong hydrogen bonds involving the guanidinium cations (N\u2014H\u22efO and N\u2014H\u22efCl) and the disordered non-coordinating water mol\u00adecule (O\u2014H\u22efO and O\u2014H\u22efCl), as well as by guanidinium \u03c0\u2013\u03c0 stacking. The structural data were confirmed by IR and UV\u2013Visible spectroscopic analysis.As part of our studies on the synthesis and the characterization of oxalate-bridged compounds A bond-valence-sum calculation, assuming Mn\u2014O and Mn\u2014Cl bonds, gives a BVS value and IDa = 0.22\u2005(4)%, respectively \u2005\u00c5] and one oxygen atom from the bridging oxalato group [Mn1\u2014O4 = 2.248\u2005(2)\u2005\u00c5]. The two oxalato groups are almost perpendicular with a dihedral angle of 89.09\u2005(6)\u00b0. The oxalate ion is located on an inversion center that also relates the two Mn atoms bonded to the oxalate ion with each other. The bridged metal ions are nearly coplanar with the oxalate plane with a mean deviation of 0.0147\u2005(8)\u2005\u00c5.The equatorial plane of the MnOII ion, as a d5 high-spin system with a spherical electron distribution, has a limited number of commonly observed coordination geometries that are based on minimization of ligand\u2013ligand repulsion. Among the Mn\u2014O distances, the shortest are those involving an oxygen atom from the oxalate ion trans to another oxygen atom from the second oxalate ion. The range of these distances is 2.180\u2005(1) to 2.194\u2005(1)\u2005\u00c5, which is in accord with those observed in other oxalate-bridged compounds such as one of the polymorphs of catena-poly[[di\u00adaqua\u00admanganese(II)]-\u03bc-oxalato-\u03ba4O1,O2:O1\u2032,O2\u2032] and 2.248\u2005(2)\u2005\u00c5.The MnII chains running along the c axis. The intra-chain Mn\u22efMn distances through bridging oxalate are 5.695\u2005(2) and 5.778\u2005(2)\u2005\u00c5, somewhat longer than the value of 5.652\u2005\u00c5 previously observed for {[Mn(C2O4)(C8H7N3)]\u00b71.5H2O}n \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z], leading to the formation of anionic layers parallel to (010). A disordered non-coordin\u00adating water mol\u00adecule acts as acceptor x, y\u00a0\u2212\u00a01, z]. Both disorder components of the non-coordinating water mol\u00adecules act as hydrogen-bond donors towards oxygen atom O3 \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a02, \u2212z]. The combined water hydrogen bonds link the anionic layers into a 3D framework.Neighbouring oxalate-bridged zigzag chains are connected with each other s Table\u00a01 towards s Table\u00a01, OW1\u2014HW2or Fig.\u00a03 for the O3 Fig.\u00a03 via the A\u22efO2i, N1\u2014H1B\u22efO4, N2\u2014H2A\u22efCl1ii, N2\u2014H2B\u22efCl1iii, N2\u2014H2A\u22efO4, N3\u2014H3A\u22efO1iv and N3\u2014H3B\u22efCl1iii , consolidating the anionic layers and giving additional stability to the three-dimensional structure as illustrated in Fig.\u00a04via \u03c0\u2013\u03c0 stacking with an inter\u00adplanar distance of 3.547\u2005(3)\u2005\u00c5 between C3 and C3 \u22121 assigned to [\u03bd(O\u2014H) + \u03bdas(NH2)] and \u03bds(NH2), respectively ], 1312 and 1409\u2005cm\u22121 [\u03bds(COO)] and 793\u2005cm\u22121 [\u03b4(COO)] et al., 2009et al., 1996n\u2192\u03c0* transition \u00b7H2O dissolved in 10\u2005cm3 of water in a 1:2:1 molar ratio. The resulting solution was left at room temperature and colourless crystals suitable for X-ray diffraction were obtained after two weeks of slow evaporation.Aqueous solutions of ammonium oxalate and guanidine hydro\u00adchloride were added to Mn using adequate HFIX instructions and refined with AFIX instructions. Hydrogen atoms of the coordinating water mol\u00adecule were found in Fourier difference maps. O\u2014H distances were restrained to a value of 0.85\u2005(1)\u2005\u00c5 and H\u22efH distances were restrained to a value of 1.387\u2005(1)\u2005\u00c5.Supra\u00admolecular features section and Fig.\u00a03The oxygen atom of the non-coordinating water mol\u00adecule had unusually high displacement parameters, and was refined as disordered over two alternative mutually exclusive positions. The solvent mol\u00adecule may be considered as being located vertically between negative-charged anionic layers formed by hydrogen-bonded polymeric chains and located horizontally between positive-charged pairs of guanidinium cations. This pseudo-channel affects its hydrogen-bonding inter\u00adactions, see the discussion in the first paragraph of the W2 and OW2B which were restrained to have similar geometries. Their hydrogen atoms were located from the Fourier difference maps. The O\u2014H bond lengths were restrained to a value of 0.85\u2005(1)\u2005\u00c5 and the H\u22efH distances were restrained to a value of 1.387\u2005(1)\u2005\u00c5. The inter\u00adatomic distances between the two pairs OW2 and HW5 and OW2B and HW3 were restrained to be equal using a SADI instruction with an effective standard deviation of 0.02. The hydrogen-bonding distance of hydrogen atom HW6 to chlorine atom Cl1 was restrained to 2.80\u2005(1)\u2005\u00c5. Subject to these and the above conditions, the occupancy ratio of the disordered non-coordinating water mol\u00adecule refined to 0.816\u2005(13):0.184\u2005(13).The disordered oxygen atom was refined as disordered over two positions O10.1107/S2056989016006605/zl2659sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016006605/zl2659Isup2.hklStructure factors: contains datablock(s) I. DOI: 1474882CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "O\u2014H\u22efO inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings organize the crystal components into columns extending along the b axis while N\u2014H\u22efN hydrogen bonds link these columns into a two-dimensional framework parallel to (100).In C 6H6N2O2\u00b7H2O, consists of N-hy\u00addroxy\u00adpicolinamide and water mol\u00adecules connected through O\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds. The O\u2014H\u22efO inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings [centroid\u2013centroid distance = 3.427\u2005(1)\u2005\u00c5] organize the components into columns extending along the b axis and the N\u2014H\u22efN hydrogen bonds link these columns into a two-dimensional framework parallel to (100). The N-hy\u00addroxy\u00adpicolinamide mol\u00adecule adopts a strongly flattened conformation and only the O\u2014H group H atom deviates significantly from the mol\u00adecule best plane. The dihedral angle between the hydroxamic group and the pyridine ring is 5.6\u2005(2)\u00b0. The conformation about the hydroxamic group C\u2014N bond is Z and that about the C\u2014C bond between the pyridine and hydroxamic groups is E.The crystal structure of the title compound, C R\u2014C(=O)\u2014NH\u2014OH. HA can exist as keto and imino\u00ad(enol) tautomers with two isomers, E and Z, for each form, and in the zwitterionic form (see Scheme below). They have found broad application in coordination chemistry due to their diversity and comparatively facile synthesis are weak organic acids with the general formula N-Hy\u00addroxy\u00adpicolinamide, known also as picoline-2-hydroxamic acid (o-PicHA), has been used extensively for the synthesis of polynuclear complexes, especially in the synthesis of diverse metallacrowns and 1.325\u2005(2)\u2005\u00c5, respectively] and a water mol\u00adecule. The N-hy\u00addroxy\u00adpicolinamide mol\u00adecule adopts a strongly flattened conformation and only the O\u2014H group H atom deviates significantly from the mol\u00adecular best plane. The maximum deviation from this plane for non-hydrogen atom is 0.083\u2005(1)\u2005\u00c5 for O1 and the hydroxyl group H2 atom is displaced from the mean plane by 0.80\u2005(1)\u2005\u00c5 in the direction of the water mol\u00adecule. The dihedral angle between the hydroxamic group and the pyridine ring is 5.6\u2005(2)\u00b0. The configuration about the hydroxamic group C\u2014N bond is Z and that about the C\u2014C bond between the pyridine and hydroxamic groups is E .The mol\u00adecular structure of the title compound is presented in Fig.\u00a01b axis while the N\u2014H\u22efN hydrogen bonds link these columns into a two-dimensional framework parallel to (100) .The mol\u00adecular components of the title compound are connected by O\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds Table\u00a01 into a tet al., 2007et al., 2001et al., 2008A search of the Cambridge Structural Database or Uiso = 1.5Ueq(O). The H atoms of the water mol\u00adecule were located in the difference Fourier maps, the O\u2014H distances standardized to 0.85\u2005\u00c5 and refined in riding-model approximation with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015024706/gk2650sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015024706/gk2650Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015024706/gk2650Isup3.cmlSupporting information file. DOI: 1444026CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the carbazole ring system is essentially planar (maximum deviation = 0.025\u2005\u00c5). The crystal packing is stabilized by inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional supra\u00admolecular network. 29H24N2, the C=N bond of the central imine group adopts an E conformation. The dihedral angles between the mean plane of the essentially planar carbazole ring system [r.m.s. deviation = 0.039\u2005(2)\u2005\u00c5] and the two phenyl rings of the 3,3-di\u00adphenyl\u00adallyl\u00adidene unit are 75.9\u2005(1) and 64.6\u2005(1)\u00b0. In the crystal, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional supra\u00admolecular network.In the title compound, C It has been reported that carbazole derivatives possess various biological activities, such as anti\u00adtumor (Itoigawa E conformation. The carbazole ring system (N1/C1\u2013C12) is essentially planar [maximum deviation = 0.039\u2005(2)\u2005\u00c5 for atom C9]. The phenyl rings C18\u2013C23 and C24\u2013C29 of the unit are oriented at dihedral angles of 75.9\u2005(1) and 64.6\u2005(1)\u00b0, respectively, to the mean plane of the carbazole ring system. The dihedral angle between the two phenyl rings is 76.1\u2005(1)\u00b0. The sum of the bond angles around atom N1 (359.7\u00b0) of the pyrrole ring is in accordance with sp2 hybridization. The geometric parameters of the title mol\u00adecule agree well with those reported for similar structures , and a methyl\u00adene H atom of the ethyl group and a benzene ring of an adjacent mol\u00adecule (C13\u2014H13A\u22efCg1iii); see Table\u00a01In the crystal, mol\u00adecules are linked by six inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional supra\u00admolecular network Table\u00a01. Four ofH-carbazol-3-amine (1\u2005mmol), 3,3-di\u00adphenyl\u00adacryl\u00adaldehyde (1\u2005mmol) and sulfated SnO2-Bi2O3-fly ash catalyst (20\u2005mg) in water (15\u2005ml) and the mixture was refluxed at 363\u2005K for 1h. On completion of the reaction (monitored by TLC with ethyl acetate and hexane as an eluent 20%) the mixture was cooled to ambient temperature. Di\u00adchloro\u00admethane (20\u2005ml) was then added to separate the organic and aqueous layers. The organic layer was filtered, dried on anhydrous Na2SO4 and the solvent removed using a rotary evaporator. The crude product obtained was purified by column chromatography on silica gel (200 mesh) with hexane and ethyl acetate (4:1) as eluent, to afford the title compound in good yield (93%). Red crystals suitable for X-ray diffraction analysis were obtained after recrystallization in CH2Cl2.A 25\u2005ml round-bottom flask was charged with 9-ethyl-9Uiso(H) = 1.5Ueq for methyl H atoms and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015005770/su5095sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015005770/su5095Isup2.hklStructure factors: contains datablock(s) I. DOI: 967497CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The hydroxide ion is surrounded by five caesium cations, which form a distorted quadratic pyramidal polyhedron. A three-dimensional network is formed by Cs\u2014Sn [3.8881\u2005(7)\u2005\u00c5 to 4.5284\u2005(7)\u2005\u00c5] and Cs\u2014NH3 [3.276\u2005(7)\u20133.636\u2005(7)\u2005\u00c5] contacts.The title compound, penta\u00adcaesium nona\u00adstannide hydroxide tetra\u00adammonia, crystallized from a solution of CsSnBi in liquid ammonia. The Sn DOI: 10.1107/S1600536814011817/ru2058Isup2.hklStructure factors: contains datablock(s) I. DOI: 1004528CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the title compound contains two crystallographically unique copper complexes. In each complex, the Cu atom is bound to two chloride ligands and to three N atoms of the 2-(4-tri\u00adfluoro\u00admethyl\u00adbenzyl\u00adidene\u00adamino)\u00adethyl\u00adamine-bis\u00ad(2\u20132amino\u00adeth\u00adyl)amine ligand to give a distorted square-based pyramidal geometry in which the axial Cu\u2014Cl bond is elongated, indicative of Jahn\u2013Teller distortion. A Cu atom from a symmetry-related mol\u00adecule is in nearby proximity to the remaining axial Cu site, thus the overall geometry about each Cu atom could be described as being between distorted square-based pyramidal and very elongated Jahn\u2013Teller-distorted octa\u00adhedral. II atom in the title compound, [CuCl2(C14H21F3N4)], adopts a coordination geometry that is between distorted square-based pyramidal and very Jahn\u2013Teller-elongated octa\u00adhedral. It is coordinated by three N atoms from the bis\u00ad(2-amino\u00adeth\u00adyl)(2-{[4-(tri\u00adfluoro\u00admeth\u00adyl)benzyl\u00adidene]amino}\u00adeth\u00adyl)amine and two chloride ligands. The two crystallographically unique copper complexes present in the asymmetric unit exhibit noticeable differences in the coordination bond lengths. Considering the CuII atoms as having square-pyramidal geometry, the basal Cu\u2014Cl bond lengths are typical [2.2701\u2005(12) and 2.2777\u2005(12)\u2005\u00c5], while the apical distances are considerably elongated [2.8505\u2005(12) and 2.9415\u2005(12)\u2005\u00c5]. For each mol\u00adecule, a CuII atom from inversion-related mol\u00adecules are in nearby proximity to the remaining axial CuII sites, but the Cu\u22efCl distances are very long [3.4056\u2005(12) and 3.1645\u2005(12)\u2005\u00c5], attributable to van der Waals contacts. Nonetheless, these contacts appear to have some structure-directing properties, leading to association into dimers. These dimers associate via stacking of the aromatic rings to form extended zigzag chains.The Cu However, an elongated Cu1\u22efCl2i distance of 3.4056\u2005(12)\u2005\u00c5 is also observed, which can be attributed to a van der Waals contact direction \u2005\u00c5 and a dihedral angle of 10.6\u2005(3)\u00b0 Fig.\u00a03. Inspecton Fig.\u00a03.i, N3\u22efCl2, N7\u22efCl4; symmetry code: (i) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z] have severely constrained N\u2014H\u22efCl angles and are merely contacts to chlorine atoms bonded to the same CuII atom. The remaining hydrogen-bond contacts are inter\u00admolecular inter\u00adactions, and while relatively long, they likely contribute to structure-directed organization.Inspection of inter\u00admolecular/intra\u00admolecular contacts reveals that amine nitro\u00adgen atoms N2, N3, N6 and N7 are involved in N\u2014H\u22efCl hydrogen-bonds Table\u00a02. HoweverN-2-bis(2-amino\u00adeth\u00adyl)amino\u00adethyl ligand skeleton and 2.609\u2005(2)\u2005\u00c5 \u00adeth\u00adyl)amine ligand: In a drybox, tris(2-(amino)\u00adeth\u00adyl)amine was dissolved in 100\u2005mL methanol in a 250\u2005mL round-bottom flask : \u03b4 2.94 , 3.70 , 7.56 , 8.08 . 13C NMR : \u03b4 55.62, 60.32, 122.85 (q), 125.73 (q), 128.35, 132.44 (q), 139.62, 160.42. FT\u2013IR (solid) v (cm\u22121): 1321 (s), 1169 (s), 1118 (s), 1062 (s), 834 (s). Melting Point: 344\u2005K. TOF\u2013ESI\u2013MS: (m/z) [M + (H)]+ calculated for C30H28N4F9 = 615.2165, found 615.2194 (4.8 p.p.m.).sk Fig.\u00a04. Ligand Synthesis of 2-(4-tri\u00adfluoro\u00admethyl\u00adbenzyl\u00adidene\u00adamino)\u00adeth\u00adyl)amine-bis(2-\u03b1minoeth\u00adyl)amine copper(II) chloride complex:tris(2-(4-Tri\u00adfluoro\u00admethyl\u00adbenzyl\u00adidene\u00adamino)\u00adeth\u00adyl)amine was dissolved in 20\u2005mL methanol in a 100\u2005mL round-bottom flask. CuCl2 was added to the flask to give a teal-colored solution. The reaction was allowed to mix for six\u2005h then 20\u2005mL of pentane was slowly added to the solution to generate a teal-colored precipitate. Solvent was removed from the round-bottom flask by connecting it to a rotary evaporator. The precipitate obtained was washed twice by transferring 15\u2005mL of pentane into the flask and stirring vigorously for thirty minutes. Solvent was removed and precipitate dried under vacuum for one\u2005h to yield a teal-colored solid . FT\u2013IR (solid): v (cm\u22121) = 1636 (m), 1506 (s), 1473 (s), 1317 (s), 1163 (s), 1109 (br), 830 (s). UV\u2013Vis (MeOH) \u03bbmax = 668\u2005nm. TOF\u2013ESI\u2013MS: (m/z) [M \u2013 2(Cl)]2+ calculated for C30H27N4F9Cu = 677.1383, found 677.1381 (0.2 p.p.m.). Blue single crystal plates suitable for X-ray analysis were obtained by slow diffusion of diethyl ether into a complex solution made in aceto\u00adnitrile at room temperature. The structure obtained is indicative of hydrolysis occuring on two amine positions of the intended copper(II) complex.Uiso(H) = 1.2Ueq for methyl\u00adene, aromatic and amide groups with C\u2014H distances set at 0.99\u2005\u00c5 (methyl\u00adene), 0.95\u2005\u00c5 (aromatic) and N\u2014H = 0.91\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989015024147/pk2570sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015024147/pk2570Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989015024147/pk2570Isup3.pdfSupporting information file. DOI: 1442779CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E conformation about the C=N bond and the mol\u00adecules is planar . In the crystal, the lattice water mol\u00adecule (Ow) links the mol\u00adecules via Ow\u2014H\u22efO, Ow\u2014H\u22efN and N\u2014H\u22efOw hydrogen bonds, forming sheets lying parallel to (100).The title compound, has an 13H10BrN3O2\u00b7H2O, the conformation about the azomethine double bond is E. The mol\u00adecule exists in the amido form with a C=O bond length of 1.229\u2005(2)\u2005\u00c5. There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond forming an S(6) ring motif. The whole mol\u00adecule is almost planar, with an r.m.s. deviation of 0.021\u2005\u00c5 for all non-H atoms, and the dihedral angle between the planes of the pyridine and benzene rings is 0.74\u2005(12)\u00b0. In the crystal, the water mol\u00adecule of crystallization links the organic mol\u00adecules via Ow\u2014H\u22efO, Ow\u2014H\u22efN and N\u2014H\u22efOw hydrogen bonds and short C\u2014H\u22efOw contacts, forming sheets lying parallel to (100). Within the sheets there is a weak \u03c0\u2013\u03c0 inter\u00adaction involving the pyridine and benzene rings [centroid-to-centroid distance = 3.8473\u2005(15)\u2005\u00c5]. The sheets are linked via C\u2014H\u22efBr inter\u00adactions, forming a three-dimensional network.In the title compound, C The two aromatic rings (C1\u2013C6 and N3/C9\u2013C13), are inclined to the almost planar hydrazone moiety by 2.12\u2005(9) and 1.40\u2005(8)\u00b0, respectively, and to each other by 0.74\u2005(12)\u00b0. There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond present in the mol\u00adecule that involves the phenolic oxygen, O1 and the azomethine nitro\u00adgen atom, N1, forming an S(6) ring motif nico\u00adtino\u00adhydra\u00adzide. The crystal structure of N\u2032-(2-hy\u00addroxy\u00adbenzyl\u00adidene)nicotinohydrazide itself is reported as a monohydrate \u00b0]. In the crystal structure of N\u2032-(2-hy\u00addroxy\u00adbenzyl\u00adidene)nico\u00adtino\u00adhydrazide monohydrate (IDASUB), the water mol\u00adecule forms three hydrogen bonds and is another example of a system where a single atom acts both as donor and acceptor.A search of the Cambridge Structural Database . Elemental analysis calculated for C13H10N3O2Br\u00b7H2O: C, 46.17, H, 3.58, N, 12.43%; found: C, 46.14, H, 3.57, N, 12.44%. IR FT\u2013IR 3059 (NH), 3269(OH), 1680 (C=O), 1584 (C=N).The title compound was prepared by adapting a reported procedure = 1.2Ueq(C). Three reflections were omitted owing to bad agreement, viz. 100, 110 and 200.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015009627/su5132sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015009627/su5132Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015009627/su5132Isup3.cmlSupporting information file. DOI: 1401825CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecule is planar, with the hy\u00addroxy H atom forming an intra\u00admolecular O\u2014H\u22efN hydrogen bond. In the crystal, mol\u00adecules form centrosymmetric dimers via two O\u2014H\u22efN hydrogen bonds. Thus, the hy\u00addroxy H atoms are involved in bifurcated O\u2014H\u22efN hydrogen bonds, leading to the formation of a central planar four-membered N2H2 ring. The dimers are bound by inter\u00admolecular \u03c0\u2013\u03c0 stacking [the shortest C\u22efC distance is 3.2997\u2005(17)\u2005\u00c5] and C\u2014H\u22ef\u03c0 inter\u00adactions into a three-dimensional framework. The crystal grown represents a new monoclinic polymorph in the space group P21/n. The mol\u00adecular structure of the present monoclinic polymorph is very similar to that of the ortho\u00adrhom\u00adbic polymorph (space group Fdd2) studied previously . The structures of the two polymorphs are distinguished by the different geometries of the hydrogen-bonded dimers, which in the crystal of the ortho\u00adrhom\u00adbic polymorph possess twofold axis symmetry, with the central N2H2 ring adopting a butterfly conformation.In an attempt to grow 8-hy\u00addroxy\u00adquinoline\u2013acetamino\u00adphen co-crystals from equimolar amounts of conformers in a chloro\u00adform\u2013ethanol solvent mixture at room temperature, the title compound, C al. 1978. Acta CrSaha 1986. Acta Cr \u00c5b = 9.243 (4) \u00c5c = 11.070 (4) \u00c5\u03b2 = 90.718 (6)\u00b0V = 677.3 (5) \u00c53Z = 4K\u03b1 radiationMo \u22121\u03bc = 0.09 mmT = 100 K0.30 \u00d7 0.25 \u00d7 0.20 mmBruker APEXII CCD diffractometerSADABS; Bruker, 2003Tmin = 0.972, Tmax = 0.981Absorption correction: multi-scan (7049 measured reflections1795 independent reflectionsI > 2\u03c3(I)1494 reflections with Rint = 0.023R[F2 > 2\u03c3(F2)] = 0.039wR(F2) = 0.109S = 1.081795 reflections103 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.39 e \u00c5\u22123\u0394\u03c1min = \u22120.20 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXTL global, I. DOI: 10.1107/S1600536814016110/rk2430Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814016110/rk2430Isup3.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S1600536814016110/rk2430fig1.tifI . DOI: I. Displacement ellipsoids are presented at the 50% probability level. H atoms are depicted as small spheres of arbitrary radius. The intra\u00admolecular O\u2014H\u22efN hydrogen bond is drawn by dashed line.Mol\u00adecular structure of Click here for additional data file.10.1107/S1600536814016110/rk2430fig2.tifI . DOI: I. The hydrogen bonds are drawn by dashed lines.The centrosymmetric H\u2013bonded dimers in the monoclinic polymorph of Click here for additional data file.10.1107/S1600536814016110/rk2430fig3.tifI . DOI: I, in which the mol\u00adecules are related by the twofold axis. The hydrogen bonds are drawn by dashed lines.The H\u2013bonded dimers in the ortho\u00adrhom\u00adbic polymorph of Click here for additional data file.10.1107/S1600536814016110/rk2430fig4.tifI . DOI: I. The hydrogen bonds are drawn by dashed lines.A portion of crystal packing of the H\u2013bonded dimers in the monoclinic polymorph of 1013310CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports5: Article number: 1580110.1038/srep15801; published online: 10302015; updated: 02022016This Article contains errors in Figure 4 and Figure 5.B/f.u.)\u2019 was incorrectly given as \u2018M(\u00b7B/f.u.)\u2019. In Figure 5, the y-axis \u2018\u2212\u0394SM (10\u22123Jule/Kg.K)\u2019 was incorrectly given as \u2018\u2212\u00b7SM (10\u22123Jule/Kg.K)\u2019. The correct Figure 4 and Figure 5 appear below as In Figure 4, the y-axis \u2018M (\u03bc"} +{"text": "The AgI atom adopts a slightly distorted linear coordination geometry. The symmetry-related right- and left-handed helical chains are arranged alternately via Ag\u22efAg and Ag\u22efF inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions, resulting in the formation of a two-dimensional supra\u00admolecular network.The reaction of AgBF L]\u00b7BF4\u00b70.5CH3OH}n, L = N-(pyridin-4-ylmeth\u00adyl)pyridine-3-amine, C11H11N3, contains one AgI ion, one ligand L, one tetra\u00adfluorido\u00adborate anion disordered over two orientations in a 0.669\u2005(13):0.331\u2005(13) ratio and one half of a methanol solvent mol\u00adecule situated on an inversion center. Each AgI ion is coordinated by two N atoms from two L ligands in a distorted linear geometry [N\u2014Ag\u2014N = 174.70\u2005(19)\u00b0]. Each L ligand bridges two AgI ions, thus forming polymeric helical chains propagating in [010]. In the crystal, Ag\u22efAg [3.3369\u2005(10)\u2005\u00c5] and \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings [centroid-to-centroid distance = 3.676\u2005(4)\u2005\u00c5] link these chains into layers parallel to (10-1). Ag\u22efF and weak N(C)\u2014H\u22efF inter\u00adactions further consolidate the crystal packing.The asymmetric unit of the title compound, {[Ag The helical chain propagates along [010] \u00b0 with respect to each other. The two pyridine rings in the L ligand are almost perpendicular, the dihedral angle between their mean planes being 89.34\u2005(15)\u00b0.The mol\u00adecular components of the title structure are shown in Fig.\u00a010] Fig.\u00a02 with a p al. 2014. The twoA = 2.84\u2005(2), Ag1\u22efF1B = 2.815\u2005(15) and Ag1\u22efF4B = 2.879\u2005(10)\u2005\u00c5] and \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings of adjacent helical chains [centroid-to-centroid distance = 3.676\u2005(4)\u2005\u00c5], resulting in the formation of a two-dimensional supra\u00admolecular network parallel to the \u2005\u00c5] and Ag\u22efF inter\u00adactions [Ag1\u22efF1ne Fig.\u00a02. Furtherne Fig.\u00a02 between et al. nitrate and perchlorate complexes of the same ligand have been reported by Zhang al. 2013. Our groN-(pyridin-4-ylmeth\u00adyl)pyridine-3-amine ligand was synthesized according to a literature method = 0.95\u2005\u00c5 for Csp2\u2014H, 0.88\u2005\u00c5 for amine N\u2014H, 0.84\u2005\u00c5 for hydroxyl O\u2014H, 0.98\u2005\u00c5 for methyl C\u2014H and 0.99\u2005\u00c5 for methyl\u00adene C\u2014H. For all H atoms Uiso(H) = 1.2\u20131.5Ueq of the parent atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901501837X/cv5498sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S205698901501837X/cv5498Isup2.hklStructure factors: contains datablock(s) I. DOI: 1428966CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Co\u2014O bond lengths in the [Co(H2O)6]2+ complex cation show very similar distances as in the related Tutton salt (NH4)2[Co(H2O)6)](SO4)2 [average 2.093\u2005(17)\u2005\u00c5], but are significantly longer than in the isotypic NiII compound (\u0394d \u2243 0.04\u2005\u00c5). The cobalt cation reaches an overall bond-valence sum of 1.97 valence units. The S\u2014O distances are nearly equal, ranging from 1.454\u2005(4) to 1.470\u2005(3)\u2005\u00c5 [mean 1.465\u2005(12)\u2005\u00c5]; however, the O\u2014S\u2014O angles vary clearly from 108.1\u2005(2) to 110.2\u2005(2)\u00b0 [average bond angle 109.5\u2005(9)\u00b0]. The non-coordinating water mol\u00adecules and di\u00admethyl\u00adammonium cations connect the sulfate tetrahedra and the [Co(H2O)6]2+ octa\u00adhedron via O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds of weak up to medium strength into a three-dimensional framework whereby the complex metal cations and sulfate anions are arranged in sheets parallel to (001).The title salt, (C DOI: 10.1107/S2056989015003400/fk2085Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015003400/fk2085fig1.tifx y z . DOI: x, \u2212y\u00a0+\u00a01, \u2212z\u00a0\u2212\u00a01.]The mol\u00adecular entities in the structure of the title compound. Displacement ellipsoids are drawn at the 50% probability level. [Symmetry code: (i) \u2212Click here for additional data file.10.1107/S2056989015003400/fk2085fig2.tif. DOI: (100)-projection of the crystal structure of the title compound. Colour scheme: S (yellow), Co (red), O (blue), N (orange), C (grey), H (colourless), H\u22efO bonds up to 1.8\u2005\u00c5 are given as red dashed lines, and from 1.85 to 2.7\u2005\u00c5 as light-blue dashed lines.1050102CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N)bis\u00ad[bis\u00ad(pyridin-2-yl-\u03baN)amine]\u00adcobalt(III)} sulfate dihydrate is comprised of discrete [Co(dpa)2(N3)2]+ cations, SO42\u2212 anions and solvent water mol\u00adecules in a 2:1:2 ratio; extensive hydrogen-bonding inter\u00adactions link the species into a three-dimensional supra\u00admolecular framework.The crystal structure of bis\u00ad{bis\u00ad(azido-\u03ba 10H9N3) as a chelating ligand, led to the synthesis and structure determination of the title compound, [Co(N3)2(dpa)2]2SO4\u00b72H2O. The crystal structure comprises discrete [Co(dpa)2(N3)2]+ cations, sulfate anions, as well as H2O solvent mol\u00adecules. The CoIII cations display a slightly distorted octa\u00adhedral coordination sphere defined by two N atoms from azide anions and four N atoms from the pyridyl rings of two dpa ligands. In the crystal, extensive C\u2014H\u22efO, N\u2014H\u22efO, and O\u2014H\u22efO inter\u00adactions result in supra\u00admolecular sheets that lie parallel to the ab plane. The sheets are further linked through O\u2014H\u22efN inter\u00adactions between the water mol\u00adecules of one sheet and azide anions of another sheet, forming a supra\u00admolecular framework.The search for new mol\u00adecular materials with inter\u00adesting magnetic properties, using the pseudohalide azide ion and di-2-pyridyl\u00adamine (dpa, C The sul\u00b0 Table\u00a01. While t0 Table\u00a01, and the0 Table\u00a01. The twoet al., 2004et al., 2004et al., 2001et al., 2005et al., 20052(N3)2]\u00b72H2O, the two pyridyl rings of each chelating dpa ligand still coordinate to the metal in a cis-disposition, but the azide anions are coordinated trans to each other [CSD refcode: XUYWIX 2]ClO4 +, the Py1cent\u2014N2\u2014Py3cent and N1\u2014Co1\u2014N3 angles are 120.92\u2005(7) and 86.48\u2005(7)\u00b0, and the Py4cent\u2014N5\u2014Py5cent and N4\u2014Co1\u2014N6 angles are 125.49\u2005(7) and 88.08\u2005(7)\u00b0 (Table\u00a01In [Co(dpa)) Table\u00a01. The C\u2014N) Table\u00a01.III cation 2(N3)2]+ complex cation inter\u00adacts with one water mol\u00adecule through a C\u2014H\u22efO hydrogen bond (C10\u2014H10\u22efO3). The sulfate anions are sandwiched between two symmetry-related layers of complex cations and water mol\u00adecules 2(N3)2]+ cations through twelve hydrogen bonds . The extensive C\u2014H\u22efO, N\u2014H\u22efO, and O\u2014H\u22efO inter\u00adactions result in two-dimensional supra\u00admolecular sheets parallel to the ab plane , forming a supra\u00admolecular framework are observed between neighboring dpa ligands and azide anions within the coordination sphere of the Con Table\u00a02. The comes Fig.\u00a04. Each suds Fig.\u00a05. As the ne Fig.\u00a05. Finallyrk Fig.\u00a06.cis\u2013trans conformation with an intra\u00admolecular C\u2014H\u22efN hydrogen bond between the two pyridyl rings \u00b7H2O with M = Mn [CSD refcode: JANPOE 2(N3)2]\u00b72H2O, the azide anions coordinate to the CuII ion weakly in a trans-fashion, resulting in a tetra\u00adgonally elongated octa\u00adhedral coordination sphere for the CuII ion, and hydrogen bonding and face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions result in two-dimensional supra\u00admolecular sheets that lie parallel to the bc-plane [CSD refcode: XUYWIX 2]ClO4 is most closely related to the title complex in that the CoIII ions are coordinated by two chelating dpa ligands and two azide anions in a cis-fashion to form [Co(dpa)2(N3)2]+ complex cations [CSD refcode: HUFNUR in water\u2013methanol . The mixture was sealed in a Teflon-lined autoclave and heated at 423\u2005K for two days and cooled to room temperature at 10\u2005K\u2005h\u22121. The crystals were obtained in ca 20% yield based on cobalt.The title compound was synthesized hydro\u00adthermally under autogenous pressure from a mixture of cobalt(II) sulfate hepta\u00adhydrate , di-2-pyridyl\u00adamine and sodium azide NaNCAUTION! Although not encountered in our experiments, azido compounds of metal ions are potentially explosive. Only a small amount of the materials should be prepared, and it should be handled with care.Uiso(H) = 1.2Ueq(C). The N\u2014H and O\u2014H-atoms were located in difference Fourier maps and then refined as riding on the carrying nitro\u00adgen or oxygen atom with Uiso(H) = 1.2Ueq(N) or Uiso(H) = 1.5Ueq(O). Two reflections considered to be affected by beam stop inter\u00adference, 0 0 2 and 2 0 0, were omitted from the refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989016003662/zl2656sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016003662/zl2656Isup2.hklStructure factors: contains datablock(s) I. DOI: 1457112CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The amide group of the ACP\u2013AZT anion is disordered (occupancy ratio 0.5:0.5), with one part forming an N\u2014H\u22efO (involving C=O\u22efH4N+) hydrogen bond and the other an O\u2014H\u22efN (involving C\u2014NH2\u22efOH2) hydrogen bond with the components of the split NH4+/H2O position. The pseudorotation parameters of ACP\u2013AZT set it apart from previously studied AZT and thymidine. In the crystal, the various components are linked by N\u2014H\u22efO, O\u2014H\u22efO, N\u2014H\u22efN, C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds, forming a three-dimensional framework.The asymmetric unit of the title compound, NH Atoms C3\u2032 and C4\u2032 deviate from the plane of atoms C1\u2032/O4\u2032/C2\u2032 by 0.458 and \u22120.101\u2005\u00c5, respectively. Unlike the AZT mol\u00adecules and the mol\u00adecule of thymidine, which exhibit a C3\u2032-exo- class of pucker, the ACP\u2013AZT mol\u00adecule exhibits a C3\u2032-endo pucker. The orientation of the thymine base relative to the de\u00adoxy\u00adribose ring in the ACP\u2013AZT mol\u00adecule is anti, similar to that in natural thymidine and AZT, the glycosyl torsion angle \u03c7ACP\u2013AZT(O4\u2032\u2014C1\u2032\u2014N1\u2014C2) = \u2212147.75\u2005(16)\u00b0. The geometric parameters of the azido residue and the orientation relative to the de\u00adoxy\u00adribose ring in ACP\u2013AZT and AZT coincide within experimental error.The mol\u00adecular structure of the title compound, ACP\u2013AZT, is illustrated in Fig.\u00a012 group of ACP\u2013AZT is disordered, one part forming a C=O\u22efH4N+ hydrogen bond and the other a C\u2014NH2\u22efOH2 hydrogen bond with the components of the NH4+/H2O position NHn Table\u00a01. In the n Table\u00a01, formingon Fig.\u00a02; howeveret al., 1986et al., 1987et al., 1987et al., 1988et al., 1988Earlier, in 1986, we studied the crystal and mol\u00adecular structures of AZT and then some other HIV replication inhibitors by X-ray analysis = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms. The other distance restraints and SIMU parameters are given below: DFIX 1.234 0.005 O7A C6\u2032 O7 C6\u2032; DFIX 0.9 N7 H7a N7 H7b; DFIX 0.95 N2S H2Sc N2S H2Sd N2S H2Sa N2S H2Sb O2S H2Sb O2S H2Sa; DFIX 1.325 0.005 N7 C6\u2032 N7A C6\u2032; DFIX 0.9 N7A H7Aa N7A H7Ab; SIMU 0.01 0.005 1.7 N2S O2S; SIMU 0.01 0.005 1.7 N7A O7 O7A N7. The split NH4+/H2O position was refined with an occupancy of 0.5 for each atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814022405/su2792sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S1600536814022405/su2792Isup2.hklStructure factors: contains datablock(s) I. DOI: 1028847CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "This induces a geometry quite different than that predicted by theory.In the crystal, mol\u00adecules pack in a centrosymmetric fashion and inter\u00adact 14H10Br2O4, the dihedral angle between the aromatic rings is 67.29\u2005(19)\u00b0. Both meth\u00adoxy-group C atoms lie close to the plane of their attached ring [deviations = \u22120.130\u2005(4) and 0.005\u2005(5)\u2005\u00c5]. In the crystal, mol\u00adecules pack in a centrosymmetric fashion and inter\u00adact via a mixture of weak \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centoid separations = 4.044\u2005(2) and 4.063\u2005(3)\u2005\u00c5], weak C\u2014H\u22efO hydrogen bonding, and Br\u22efBr halogen bonding. This induces a geometry quite different than that predicted by theory.In the title compound, C This effect is rationalized as a loss of overlap between two \u03c0 systems. Inter\u00adrupting conjugation is a prerequisite for the design of unimolecular rectifiers \u00b0 in the crystal structure. DFT (B3LYP-DGDZVP) calculations performed on the target mol\u00adecule in the gas phase predict an angle of \u221238.54\u00b0. This significant discrepancy is probably due to packing inter\u00adactions in the solid phase.ortho to these positions. Finally, the quinone ring is slightly buckled (r.m.s. deviation = 0.064\u2005\u00c5), probably due to supra\u00admolecular packing effects.Substituents on the HBQ system behave as expected. The C\u2014Br bond distances reflect the natures of the electron-deficient quinone and electron-rich di\u00admeth\u00adoxy\u00adbenzene rings: the C1\u2014Br1 bond distance is 1.872\u2005(5)\u2005\u00c5, while the C10\u2014Br2 bond is 1.897\u2005(4)\u2005\u00c5. Thus Br1 has a slightly stronger \u03c0-donating character into the quinone moiety, strengthening the bond relative to the C10\u2014Br2 bond of the di\u00admeth\u00adoxy\u00adbenzene ring. The meth\u00adoxy substituents are nearly coplanar to the benzene ring, with a C12\u2014C11\u2014O4\u2014C14 torsion angle of 1.5\u2005(6)\u00b0 and a C9\u2014C8\u2014O3\u2014C13 torsion angle of \u22124.4\u2005(5)\u00b0. The methyl portions of each of these groups point away from the sterically restricting groups c axis show some face-to-face \u03c0-stacking.Each mol\u00adecule is surrounded by eight neighboring mol\u00adecules, which inter\u00adact through hydrogen bonding, halogen bonding, and \u03c0\u2013\u03c0 inter\u00adactions Figs. 2 and 3 \u25b8.a axis, the benzene rings \u2018nestle\u2019 closely to one another in an anti\u00adparallel geometry, where one quinone points up and the layer behind it points down. Within the cb plane, the benzene rings are coplanar; hydrogen atoms from C14 on one mol\u00adecule project closely to O3 on the adjacent mol\u00adecule and vice versa for a hydrogen atom attached to C13 to the adjacent O4 . The Br1\u22efBr2 separation is 3.4204\u2005(8)\u2005\u00c5, with almost linear C1\u2014Br1\u22efBr2 and C10\u2014Br2\u22efBr1 angles of 178.2\u2005(4) and 170.9\u2005(4)\u00b0, respectively. Equivalent rings from mol\u00adecules packed along this axis are parallel to one another; the quinone and benzene rings aligned coplanar to the corresponding ring in the next mol\u00adecule.Mol\u00adecules are aligned linearly in a head-to-tail manner where the bromine atoms participate in Br\u22efBr halogen bonding Fig.\u00a03. As disc2O. A solution of 2-bromo-1,4-di\u00admeth\u00adoxy\u00adbenzene in 25\u2005ml of aceto\u00adnitrile was quickly added with vigorous stirring. After three hours, the product had precipitated as a grey\u2013green powder. The precipitate was filtered, washed with water, and dried. The crude product was purified using flash chromatography , yielding 0.0959\u2005g of the desired product (20.3%). Crystals were obtained by slow evaporation of a solution in chloro\u00adform.Cerium(IV) ammonium nitrate was dissolved in 30\u2005ml of HUiso(H) = 1.5Ueq(C) for methyl H atoms, C\u2014H = 0.95\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for other H atoms]. Hydrogen atoms on methyl groups were refined with a riding rotating model. Crystal data, data collection and structure refinement details are summarized in Table\u00a02Hydrogen atoms were placed in calculated positions, and their coordinates and displacement parameters were constrained to ride on the carrier atom [C\u2014H = 0.98\u2005\u00c5 and 10.1107/S2056989015020472/hb7514sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989015020472/hb7514Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015020472/hb7514Isup3.cmlSupporting information file. DOI: 1433845CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A receptors are the main inhibitory neurotransmitter receptors in the brain and are targets for numerous clinically important drugs such as benzodiazepines, anxiolytics and anesthetics. We previously identified novel ligands of the classical benzodiazepine binding pocket in \u03b11\u03b22\u03b32 GABAA receptors using an experiment-guided virtual screening (EGVS) method. This screen also identified novel ligands for intramembrane low affinity diazepam site(s). In the current study we have further characterized compounds 31 and 132 identified with EGVS as well as 4-O-methylhonokiol. We investigated the site of action of these compounds in \u03b11\u03b22\u03b32 GABAA receptors expressed in Xenopus laevis oocytes using voltage-clamp electrophysiology combined with a benzodiazepine site antagonist and transmembrane domain mutations. All three compounds act mainly through the two \u03b2+/\u03b1\u2212 subunit transmembrane interfaces of the GABAA receptors. We then used concatenated receptors to dissect the involvement of individual \u03b2+/\u03b1\u2212 interfaces. We further demonstrated that these compounds have anesthetic activity in a small aquatic animal model, Xenopus laevis tadpoles. The newly identified compounds may serve as scaffolds for the development of novel anesthetics.GABA A) receptor. These receptors are composed of five homologous subunits organized around a central Cl\u2212 selective channelA receptor have been cloned , integrating experimental data with homology modeling of the GABAin vivo, revealing potencies similar to propofol.Here we describe the actions of two compouds identified by EGVS, 31 and 132A receptors we screened 198 compounds for displacement of the high affinity benzodiazepine site antagonist [3H]-Ro 15-1788 at receptors expressed in HEK-cells. Many high affinity ligands were identified3H]-Ro 15-1788 from site 1 but potently enhanced GABAA receptor activation. 4-O-methylhonokiol shares these characteristics, potentiating \u03b11\u03b22\u03b32 receptors with an EC50 of 5.4\u2009\u00b1\u20091.8\u2009\u03bcM, independent of the high affinity site for benzodiazepinesIn an attempt to find novel ligands for the high affinity site for benzodiazepines on GABA1\u03b22\u03b32 GABAA receptors expressed in Xenopus oocytes. These compounds elicited at the concentration of 3\u2009\u03bcM and 30\u2009\u03bcM currents amounting to 0.1\u2009\u00b1\u20090.06% and 0.3\u2009\u00b1\u20090.13% , respectively of the maximal current amplitude elicited by GABA in the same oocytes. Thus neither of the compounds tested acts as an appreciable agonist on \u03b11\u03b22\u03b32 receptors.First, we investigated if compounds 31 and 132 were able to act as agonists. 1\u03b22\u03b32 receptors. For compound 31, at high concentrations apparent desensitization was observed, that could be partly due to open channel blocker effect. For both compounds no saturation at the highest concentration was obtained, because of poor solubility we could not test higher concentrations.Both compounds strongly enhanced currents elicited by GABA. We established the concentration response curves of this positive allosteric modulation. After two applications of GABA at a concentration eliciting 0.5\u20131.5% of the maximal current amplitude, the same concentration of GABA was co-applied with increasing concentrations of the tested compounds. 1\u03b22\u03b32 receptors. Nevertheless, we tested whether 1\u2009\u03bcM of the site 1 antagonist Ro 15-1788 inhibits potentiation of GABA currents by compounds 31 or 132. Either compound (3\u2009\u03bcM) strongly potentiated currents elicited by GABA. Potentiation by either drug was not inhibited by 1\u2009\u03bcM Ro 15-1788 (t test) for compound 31, and 115\u2009\u00b1\u200914% for compound 132. This confirms that potentiation by compounds 31 and 132 does not result from action at the benzodiazepine site 1.From the binding data we did not expect that the two compounds are acting through the high affinity benzodiazepine binding site 1 in \u03b1 15-1788 . Relativ1\u03b22\u03b32 receptors, \u03b11S269I, \u03b22N265I and \u03b32S280I, eliminated the potentiation by high concentrations of diazepam1M, \u03b22M and \u03b32M. Recently, we described the potency of GABA to activate all of these receptor subtypes expressed in Xenopus oocytes3 receptor3 subunits were renamed \u03b11, \u03b22 and \u03b32 , and by compound 132 to 118\u2009\u00b1\u200944% . Modulation in \u03b11\u03b22\u03b32M receptors by compound 31 was 87\u2009\u00b1\u200952% , and by compound 132 was 91\u2009\u00b1\u200921% . In contrast, modulation by both compounds was strongly impaired in \u03b11\u03b22M\u03b32 and triply mutated \u03b11M\u03b22M\u03b32M receptors. Potentiation by 3\u2009\u03bcM compound 31 in \u03b11\u03b22M\u03b32 was 10.4\u2009\u00b1\u20098.5% , and in \u03b11M\u03b22M\u03b32M receptors was 2.3\u2009\u00b1\u20090.8% . Potentiation by compound 132 relative to that in \u03b11\u03b22\u03b32 was also dramatically reduced in \u03b11\u03b22M\u03b32 7.3\u2009\u00b1\u20096.8% and \u03b11M\u03b22M\u03b32M \u22120.4\u2009\u00b1\u20095.1% , respectively.1\u03b22M\u03b32 and \u03b11M\u03b22M\u03b32M receptors, similar to compounds 31 and 132. Relative to wild-type \u03b11\u03b22\u03b32 receptors, modulation in \u03b11M\u03b22\u03b32 mutated amounted to 63\u2009\u00b1\u20096% , and in \u03b11\u03b22\u03b32M receptors to 79\u2009\u00b1\u200916% . Relative to \u03b11\u03b22\u03b32, residual potentiation in \u03b11\u03b22M\u03b32 receptors amounted to 15.6\u2009\u00b1\u20091.5% , and to 2.2\u2009\u00b1\u20090.2% in \u03b11M\u03b22M\u03b32M.Potentiation by 1\u2009\u03bcM 4-O-methylhonokiol was also significantly reduced only in \u03b11\u03b22\u03b32 GABAA receptors.The above data suggests that the modulatory site(s) for the three compounds we studied is located in one or both of the \u03b2+/\u03b1\u2212 TMD subunit interfaces on \u03b1A receptor contains two \u03b2+/\u03b1\u2212 subunit interfaces. Combined mutation at these interfaces greatly reduces the modulatory effects of compounds 31, 132 and 4-O-methylhonokiol. Using \u03b11-\u03b22-\u03b11 and \u03b32-\u03b22 subunit concatemers, we studied the effects of individual mutated interfaces. We designated receptors containing the mutant in the \u03b32-\u03b22 construct interface 1 M, and the mutation in the \u03b11-\u03b22-\u03b11 construct interface 2 M potentiation for all three compounds was abolished. Relative to \u03b11-\u03b22-\u03b11/\u03b32-\u03b22 concatenated receptors, in the double mutant \u03b11-\u03b22M-\u03b11/\u03b32-\u03b22M receptor modulation by compound 31 was reduced to 1\u2009\u00b1\u20095% , by compound 132 to \u22122\u2009\u00b1\u20090.4% , and by 4-O-methylhonokilol to 1\u2009\u00b1\u20090.6% . In receptors containing only one mutation, either interface 1 or interface 2, modulation by compound 31 was reduced significantly compared to concatenated wild type receptors \u03b11-\u03b22-\u03b11/\u03b32-\u03b22. With interface 1 M, residual relative potentiation was 28\u2009\u00b1\u20096% and with interface 2\u2009M it was 51\u2009\u00b1\u20093% of wild-type. Potentiation of interface 1\u2009M and interface 2\u2009M receptors differed significantly . Therefore, both TMD \u03b2+/\u03b1\u2212 sites seem to contribute differently to modulation by compound 31, interface 1 being more efficacious.Wild type concatenated receptors \u03b1Tukey posthoc test), and in interface 2\u2009M receptors to 39\u2009\u00b1\u200919% . Again, interface 1\u2009M produced a larger impact than 2\u2009M, although the difference was at the statistical limit . Modulation by 4-O-methylhonokiol in interface 1\u2009M and 2\u2009M receptors was reduced to 11\u2009\u00b1\u20096% and 28\u2009\u00b1\u20098% , respectively. The two mutations produced significantly different effects , with the interface 1\u2009M effect again larger.Likewise, modulation by compound 132 was sensitive to the mutations at both sites. In interface 1 M receptors relative potentiation was reduced to 8\u2009\u00b1\u20094% , to 125\u2009\u00b1\u200925% in the mutated receptor . In contrast, this mutation did not significantly reduce potentiation by 3\u2009\u03bcM compound 132: 415\u2009\u00b1\u2009126% in wild-type receptors (mean\u2009\u00b1\u2009SD n\u2009=\u20096) versus 280\u2009\u00b1\u200936% in the mutated receptor . We have shown earlier that potentiation by 4-O-methylhonokiol in receptors carrying the \u03b22N265S mutation was greatly reduced to about 40%The \u03b21 subunit by different \u03b1 subunit isoforms: \u03b11\u03b22\u03b32, \u03b12\u03b22\u03b32, \u03b13\u03b22\u03b32, \u03b14\u03b22\u03b32, \u03b15\u03b22\u03b32 and \u03b16\u03b22\u03b32 , \u03b12\u03b22\u03b32 receptors, 407\u2009\u00b1\u2009172% , \u03b13\u03b22\u03b32 receptors, 735\u2009\u00b1\u2009234%, , and \u03b14\u03b22\u03b32 receptors, 412\u2009\u00b1\u2009175% . The \u03b15\u03b22\u03b32 receptor showed a significant decrease in potentiation compared to \u03b11\u03b22\u03b32 receptors to 253\u2009\u00b1\u200992% , and the \u03b16\u03b22\u03b32 receptor an increase in potentiation, amounting to 780\u2009\u00b1\u2009236% . This discrepancy in modulation of \u03b15\u03b22\u03b32 and \u03b16\u03b22\u03b32 receptors maybe explained by the fact that 3 of the first 7 residues of M1 located at the minus side of the \u03b1 subunit are different in \u03b12\u03b22\u03b32 receptors, 652\u2009\u00b1\u2009215% in \u03b13\u03b22\u03b32 receptors, 476\u2009\u00b1\u2009152% in \u03b14\u03b22\u03b32 receptors, 399\u2009\u00b1\u2009147% in \u03b15\u03b22\u03b32 receptors, and 542\u2009\u00b1\u2009175% in \u03b12\u03b22\u03b32 receptor type. These results indicate that although the type of \u03b1 subunit has differential effects on potentiation between compounds 31 and 132, these compounds modulate all receptor subtypes studied.From the above experiments we inferred that the \u03b2+/\u03b1\u2212 TMD subunit interfaces mediate potentiation of compounds 31 and 132. We also wanted to know if potentiation by these compounds depends on subunit isoforms. First we replaced the \u03b1d \u03b16\u03b22\u03b32 . Compounifferent . Compoun2 by \u03b21 or \u03b23. For compounds 31 and 132, \u03b11\u03b23\u03b32 receptors showed a similar potentiation as \u03b11\u03b22\u03b32 receptors. Amounting to 381\u2009\u00b1\u2009115% for compound 31 and 555\u2009\u00b1\u2009169% for compound 132 . In the case of \u03b11\u03b21\u03b32 receptors, potentiation by both compounds was significantly reduced compared to that in \u03b11\u03b22\u03b32, 41\u2009\u00b1\u200913% for compound 31 and 37\u2009\u00b1\u20097% for compound 132 . These results indicate that the type of \u03b2 subunit is important for the potentiation by both compounds. It is interesting to note in this context that \u03b21 and \u03b22/\u03b23 differ not only in the residue 265, but also in the fourth residue of M3 predicted to be close to the latter residue . Compound 132 also showed a similar potentiation between both receptors, potentiation in \u03b11\u03b22 receptors amounting to 625\u2009\u00b1\u2009219% . When the \u03b32 subunit was replaced by a \u03b4 subunit, in the case of \u03b11\u03b22\u03b4 receptors, potentiation was affected only in the case of compound 31, where a significant reduction was observed relative to \u03b11\u03b22\u03b32 receptors. Where potentiation amounted to 168\u2009\u00b1\u200948% for compound 31, and 640\u2009\u00b1\u2009275% for compound 132. In \u03b14\u03b23\u03b4 receptors, potentiation by both compounds was statistically reduced relative to \u03b14\u03b23\u03b32 receptors. For compound 31 potentiation amounted to 345\u2009\u00b1\u200977% in \u03b14\u03b23\u03b32 receptors, and was reduced to 125\u2009\u00b1\u200922% in \u03b14\u03b23\u03b4 receptors. Potentiation of compound 132 in \u03b14\u03b23\u03b32 receptors was 728\u2009\u00b1\u2009203% , and was reduced to 214\u2009\u00b1\u200971% in \u03b14\u03b23\u03b4 receptors.When the \u03b35 and \u03b16 subunits. The type of \u03b2 subunit was also important, as the presence of the \u03b21 subunit strongly reduced potentiation by this compound. On the contrary, the presence of a \u03b3 or a \u03b4 subunit did not affect potentiation.Previous workXenopus tadpoles, 50 of 2.7\u2009\u03bcM , while the EC50 compound 132 was 1.2\u2009\u03bcM , and 4-O-methylhonokiol EC50\u2009=\u20091.0\u2009\u03bcM . For comparison, the EC50 for propofol-induced LoRR in tadpoles is 1.3\u2009\u03bcMThe anesthetic activity for compounds 31, 132 and 4-O-methylhonokiol was determined as loss of righting reflex (LoRR) in 1\u03b22\u03b32 GABAA receptors that do not act at site 1. We suspected that they instead act through the low affinity TMD site(s) for benzodiazepines (site 3). Indeed, potentiation by compounds 31, 132 and 4-O-methylhonokiol was abolished in the triple mutant receptor \u03b11S269I\u03b22N265I\u03b32S280I as well as by the single mutation \u03b22N265I, but unaffected by the homologous mutations \u03b11S269I and \u03b32S280I. Assuming that these TM2 mutations alter drug actions through local steric effects in adjacent TMD interfacial sites, our results indicate that of the five such sites, only the two \u03b2+/\u03b1\u2212 interfaces 1 and 2 mediate the potentiating effects of these three compounds.Here we functionally characterized compounds 31, 132 and further investigated the properties of 4-O-methylhonokiol. All three compounds are potent allosteric potentiators of \u03b1We further dissected the contribution of the individual \u03b2+/\u03b1- subunit interfaces using concatenated subunit assemblies. The \u03b3\u03b2+/\u03b1\u2212\u03b2 interface (interface 1) and \u03b1\u03b2+/\u03b1\u2212\u03b3 interface (2) participated differently in modulation by the three compounds studied. For all compounds the contribution of the interface 1 to drug modulation is apparently greater than that of the interface 2.1\u03b22\u03b32 receptors, \u03b22N265I reduced potentiation by all compounds, \u03b11S269I reduced potentiation exclusively by pentobarbital, and the \u03b32S280I mutation increased potentiation by etomidate, while reducing potentiation by propofol and pentobarbital23243342We and others have shown that the intravenous anesthetics etomidate, propofol and pentobarbital also act via TMD interfacial sites262728292M286W) and different concatenated subunit assemblies suggest that etomidate interactions are equivalent in the two \u03b2+/\u03b1\u2212 sites of \u03b11\u03b22\u03b32 receptorsWork by our group using receptor concatenation determined that both \u03b2+/\u03b1\u2212 subunit interfaces participated equally in modulation by propofol. In contrast, modulation by etomidate was found to be more affected by the \u03b3\u03b2+/\u03b1\u2212\u03b2 interface site (interface 1) than the \u03b1\u03b2+/\u03b1\u2212\u03b3 site (interface 2)2N265 in \u03b21 and \u03b23 are serine and asparagine, respectively, and this single residue dramatically influences sensitivity to loreclezole262829391\u03b21\u03b32 receptors, compared to \u03b11\u03b22\u03b32 receptors. Potentiation by 4-O-methylhonokiol is also reduced by the \u03b22N265S mutation or substitution of \u03b21 for \u03b22The homologs of \u03b24 and \u03b16 subunits, contrasting with benzodiazepine site 1 agonists. Both compounds also potentiated receptors carrying the \u03b4 subunit, although to different degrees. Thus, these compounds not only acted at receptors shown to be located synaptically as \u03b11\u03b22\u20133\u03b32, \u03b12\u03b22\u03b32 and \u03b13\u03b22\u03b325\u03b22\u03b32 receptors and receptors containing the \u03b4 subunit, which are all located extra-synaptically546In subunit specificity studies, compounds 31 and 132 potentiated receptors containing the \u03b1Xenopus laevis tadpoles with EC50s comparable to the anesthetics propofolThe similarities between the three compounds we studied and the clinical anesthetics propofol, etomidate and pentobarbital suggested their possible use as sedative-hypnotics in animals. Indeed, all three compounds induced reversible loss of righting reflexes (LoRR) in A receptors at molecular sites different from the classical benzodiazepine pocket. These compounds, together with 4-O-methylhonokiol, act through \u03b2+/\u03b1\u2212 TMD interfaces, with strongest effects through the interface 1. These compounds potently produced LoRR in aquatic animals and thus may be useful lead compounds in the search for novel anesthetic, sedative-hypnotic or anxiolytic drugs.In summary, the newly identified compounds 31 and 132 modulate both synaptic and extrasynaptic GABA1S269I, \u03b22N265I, \u03b22N265S and \u03b32S280I were prepared using the QuikChangeTM mutagenesis kit .The point mutations \u03b12-\u03b22, and triple construct \u03b11-\u03b22-\u03b11 has been described previously2N265 to I was done in the tandem construct and the triple construct using the QuikChangeTM mutagenesis kit .Construction of tandem and triple subunit cDNAs. The tandem construct \u03b3Capped cRNAs were synthesized from the linearized plasmids with a cytomegalovirus promotor (pCMV vectors) containing the different subunits, respectively. A poly-A tail of about 400 residues was added to each transcript using yeast poly-A polymerase . The concentration of the cRNA was quantified on a formaldehyde gel using Radiant Red stain (Bio-Rad) for visualization of the RNA. Known concentrations of RNA ladder (Invitrogen) were loaded as standard on the same gel. cRNAs were precipitated in ethanol/isoamylalcohol 19:1, the dried pellet dissolved in water and stored at \u221280\u2009\u00b0C. cRNA mixtures were prepared from these stock solutions and stored at \u221280\u2009\u00b0C.1, \u03b22 and \u03b32 subunits of the GABAA receptors at a concentration of 10 nM:10 nM:50 nMAnimal experiments were carried out in strict accordance to the Swiss ethical guidelines, and have been approved by the local committee of the Canton Bern Kantonstierarzt, Kantonaler Veterin\u00e4rdienst Bern (BE85/15). Surgery was done under anesthesia, and all efforts were made to diminish animal suffering. Xenopus laevis oocytes were prepared, injected and defolliculated as described previously492, 1\u2009mM CaCl2, and 5\u2009mM Na-HEPES (pH 7.4). Concentration response curves for the compounds were fitted with the equation I(c)\u2009=\u2009Imax/[1 + (EC50/c)n], where c is the concentration of the compound, EC50 the concentration eliciting half-maximal current amplitude, Imax is the maximal current amplitude, I the current amplitude, and n is the Hill coefficient. Maximal current amplitudes (Imax) were obtained from the fits of the concentration-response curves. For all receptors studied, modulation was measured at a GABA concentration eliciting 0.5\u20131.5% of the maximal GABA current amplitude. GABA was applied twice alone for 20\u201360\u2009s, and then in combination with the different compounds for 45\u2009s or 1\u2009min. The duration of washout periods was 4\u2009min in between agonist or agonist/drug aplications to prevent receptor desentization. At the beginning of the experiments, GABA applications were repeated when the elicited current amplitude altered by >5%. Potentiation was calculated by the following equation: (IModulator + GABA/IGABA\u2009\u2212\u20091) * 100%. The perfusion solution was applied through a glass capillary with an inner diameter of 1.35\u2009mm, the mouth of which was placed about 0.4\u2009mm from the surface of the oocyte. This allowed fast changes in agonist concentration around the oocyte. The rate of change was estimated 70% in less than 0.5\u2009sCurrents were measured using a modified two-electrode voltage clamp amplifier Oocyte clamp OC-725 (Warner Instruments) in combination with a XY-recorder (90% response time 0.1\u2009s) or digitized at 100\u2009Hz using a PowerLab 2/20 (AD Instruments) using the computer programs Chart . Tests with a model oocyte were performed to ensure linearity in the larger current range. The response was linear up to 15\u2009\u03bcA. The holding potential was \u221280\u2009mV. The perfusion medium contained 90\u2009mM NaCl, 1\u2009mM KCl, 1\u2009mM MgClt test was used to compare two means. One-way analysis of variance (ANOVA) was used for multiple comparisons followed by a Tukey post hoc test. *p\u2009<\u20090.05; **p\u2009<\u20090.01; ***p\u2009<\u20090.001.All data are from at least two different batches of oocytes. Data represent mean\u2009\u00b1\u2009SD. An unpaired Xenopus laevis tadpoles as previously described415250\u2009\u2212\u2009Log[Drug])*HillSlope)) using Graphpad Prism 6. Results are reported as EC50s and 95% confidence intervals.Animals were used and experiments were carried out with approval and according to the guidelines of the MGH Institutional Animal Care and Use Committee. General anesthetic potency was assessed in How to cite this article: Maldifassi, M. C. et al. Novel positive allosteric modulators of GABAA receptors with anesthetic activity. Sci. Rep.6, 25943; doi: 10.1038/srep25943 (2016)."} +{"text": "The mol\u00adecule is arranged around a center of symmetry with half the crown ether mol\u00adecule and one mol\u00adecule of aceto\u00adnitrile symmetry independent. All O\u2014C\u2014C\u2014O torsion angles are gauche while all C\u2014O\u2014C\u2014C angles are trans. The sequence of torsion angles is [(+ttg)(\u2212ttg)]3; the geometry of oxygen atoms is close to pseudo-Dd3 with three atoms below and three atoms above the mean plane, with an average deviation of \u00b10.16\u2005(1)\u2005\u00c5 from the mean plane. This geometry is identical to that observed in metal ion complexes of di\u00adcyclo\u00adhexane-18-crown-6 but differs significantly from the conformation of a free unsolvated mol\u00adecule. Each aceto\u00adnitrile mol\u00adecule connects to a crown ether mol\u00adecule via two of its methyl group H atoms (C\u2014H\u22efO). Weaker inter\u00adactions exist between the third H atom of the aceto\u00adnitrile methyl group and an O atom of a neighbouring crown ether mol\u00adecule (C\u2014H\u22efO); and between the N atom of the aceto\u00adnitrile mol\u00adecule and a H atom of another neighbouring crown ether mol\u00adecule. All these inter\u00admolecular inter\u00adactions create a three-dimensional network stabilizing the disolvate.The title compound (systematic name: \u00c5b = 9.5286 (5) \u00c5c = 9.8927 (6) \u00c5\u03b1 = 80.415 (2)\u00b0\u03b2 = 81.697 (2)\u00b0\u03b3 = 80.927 (2)\u00b0V = 632.53 (6) \u00c53Z = 1K\u03b1 radiationMo \u22121\u03bc = 0.09 mmT = 173 K0.49 \u00d7 0.34 \u00d7 0.28 mmBruker PHOTON-100 CMOS diffractometerSADABS; Bruker, 2014Tmin = 0.956, Tmax = 1.000Absorption correction: multi-scan (20203 measured reflections3045 independent reflectionsI > 2\u03c3(I)2476 reflections with Rint = 0.031R[F2 > 2\u03c3(F2)] = 0.041wR(F2) = 0.104S = 1.073045 reflections229 parametersAll H-atom parameters refinedmax = 0.29 e \u00c5\u22123\u0394\u03c1min = \u22120.19 e \u00c5\u22123\u0394\u03c1APEX2 used to solve structure: SHELXT I. DOI: 10.1107/S2056989015011056/zl2627Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015011056/zl2627Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015011056/zl2627Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015011056/zl2627fig1.tifx z . DOI: x, 1 \u2212 y, 1\u00a0\u2212\u00a0z).Structure of the title compound with atom labeling. The second half of the crown ether mol\u00adecule and the second aceto\u00adnitrile mol\u00adecule mol\u00adecule are created by an inversion center located at the center of the crown ether mol\u00adecule (symmetry operator: 1\u00a0\u2212\u00a0Click here for additional data file.10.1107/S2056989015011056/zl2627fig2.tif. DOI: Inter\u00admolecular short contacts of aceto\u00adnitrile mol\u00adecules with neighboring crown ether mol\u00adecules.1405283CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The coordinating water mol\u00adecule is located on a twofold rotation axis while the L2\u2212 anion sits about an inversion center. The L2\u2212 anions bridge the CuII cations, forming polymeric chains propagating along the [101] direction. In the crystal, O\u2014H\u22efO, N\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22ef\u03c0 inter\u00adaction link the polymeric chains and the solvent water mol\u00adecules into a three-dimensional supra\u00admolecular architecture.In the title polymeric complex, {[Cu(C DOI: 10.1107/S205698901500626X/xu5844Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901500626X/xu5844fig1.tifx y z x y z . DOI: x, y, \u2212z\u00a0+\u00a0x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a01]The mol\u00adecular structure of (I), a drawing of the asymmetric unit (multi-colored portion) with displacement ellipsoids at the 30% probability level. [symmetry code: (i) 1\u00a0\u2212\u00a0Click here for additional data file.10.1107/S205698901500626X/xu5844fig2.tif. DOI: The polymeric chain of (I).1056415CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The geometry around the cationic PdII centre is distorted square planar, chelated by the imine N- and phenolate O-donor atoms of the two Schiff base ligands. The N- and O-donor atoms of the two ligands are mutually trans, with Pd\u2014N and Pd\u2014O bond lengths of 2.028\u2005(2) and 1.9770\u2005(18)\u2005\u00c5, respectively. The fluoro\u00adphenyl ring is tilted away from the coordination plane and makes a dihedral angle of 66.2\u2005(2)\u00b0 with the phenolate ring. In the crystal, mol\u00adecules are linked into chains along the [101] direction by weak C\u2014H\u22efO hydrogen bonds. Weak \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances of 4.079\u2005(2)\u2005\u00c5 stack the mol\u00adecules along c.The asymmetric unit of the title complex, [Pd(C II and NiII complexes of Schiff bases have attracted much attention as they play important roles in bioinorganic chemistry and may provide the basis for models of active sites of biological systems 2], is related to the previously reported compound bis\u00ad{2-[(E)-(4-fluoro\u00adbenz\u00adyl)imino\u00admeth\u00adyl]phen\u00ado\u00adlato-\u03ba2N,O1}nickel(II) was screened for catalytic activity in the Suzuki cross-coupling reaction between phenyl\u00adboronic acid and iodo\u00adbenzene with a catalyst loading of 1\u2005mmol%. The conversion of iodo\u00adbenzene was found to occur with a yield of 52%.II cation lying on an inversion centre and the Schiff base anion acting as an N,O bidentate chelate ligand \u2005\u00c5, \u03b8 = 112.8\u2005(3) and \u03d5 = 206.9\u2005(3)\u00b0. Other bond lengths and angles observed in the structure are also normal. The fluoro\u00adphenyl ring (C1\u2013C6) makes a dihedral angle of 66.2\u2005(2)\u00b0 with the phenolate ring (C9\u2013C14).The asymmetric unit of (1) contains one-half of the mol\u00adecule with the Pdnd Fig.\u00a01. The PdIs Table\u00a01 are typiA\u22efO1 inter\u00adactions \u2005\u00c5 . These combine with the C\u2014H\u22efO contacts to generate sheets in the ac plane , the mol\u00adecules are linked into chains along the [101] direction by weak C4\u2014H4ns Fig.\u00a02. A weak ns Fig.\u00a02, with a ne Fig.\u00a04. These sII complexes with related Schiff base N2O2 donor sets have been reported -2-(1-(4-fluoro\u00adbenzyl\u00adimino)\u00adeth\u00adyl)phenol in a round-bottomed flask. Palladium(II) acetate was dissolved separ\u00adately in CH3CN (20\u2005mL) and was then added into the flask containing the ligand solution. The mixture was stirred and refluxed for 5\u2005h upon which a turmeric yellow solid was formed. The solid was filtered off, washed with ice-cold CH3CN and air dried at room temperature. The solid product was recrystallized from chloro\u00adform, yielding yellow crystals (yield 48.5%). 1H NMR, 13C NMR and IR spectral bands have been studied and agree well with the structure obtained from the values of the CHN analyses and X-ray structure determination.The ligand, : 1598 \u03bd(C=N), 1319 \u03bd(C\u2014N), 1216 \u03bd(C\u2014O), 1321 \u03bd(CH3), 556 \u03bd(Pd\u2014N), 450 \u03bd(Pd\u2014O). 1H NMR : \u03b4 (p.p.m.) 2.32 , 5.11 , 6.53\u20137.46 (ArC). 13C NMR : \u03b4 (p.p.m.) 19.5 (C\u2014CH3), 54.2 (CH2), 115.3, 115.6, 121.3, 128.6, 130.2 (ArC), 169.8 (C=N).Melting point 508\u2013510\u2005K. Analytical data for C\u22121 assignable to the C=N stretching frequency of the azomethine moiety. Weak bands at 556 and 450\u2005cm\u22121 attributable to Pd\u2014N and Pd\u2014O vibrations, respectively as that observed by Gupta et al. (20132) and methyl (\u2013CH3) protons appear at 5.11 and 2.32 p.p.m., respectively. The 13C chemical shift for the imine carbon (C=N) is found at 169.8 p.p.m., agreeing with data reported by \u015eenol et al. (2011The infrared spectrum of (1) exhibits a strong band at 1598\u2005cm al. 2013. Singlet al. 2011.The title compound was screened for catalytic activity in the Suzuki cross-coupling reaction between phenyl\u00adboronic acid with iodo\u00adbenzene. The reaction was carried out under nitro\u00adgen at 373\u2005K in di\u00admethyl\u00adacetamide with a catalyst loading of 1\u2005mmol%. The conversion of iodo\u00adbenzene was monitored using GC\u2013FID after 24\u2005hours of reaction time. This resulted in a 52% conversion of iodo\u00adbenzene in the reaction.d(C\u2014H) = 0.93\u2005\u00c5 for aromatic, 0.97\u2005\u00c5 for CH2 and 0.96\u2005\u00c5 for CH3 hydrogen atoms. The Uiso values were constrained to be 1.5Ueq of the carrier atom for methyl H atoms and 1.2Ueq for the remaining H atoms. A rotating group model was used for the methyl groups.Crystal data, data collection and crystal structure refinement details are summarized in Table\u00a0310.1107/S2056989015004405/sj5444sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015004405/sj5444Isup2.hklStructure factors: contains datablock(s) I. DOI: 1045879CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular conformation and crystal packing are governed by a network of hydrogen bonds and by \u03c0\u2013\u03c0 stacking.The crystal structure of the title compound allowed the 14H15ClN2O4, prepared by reaction of a methacryloyl dimer with nitro\u00adaniline, was determined to establish the relative substituent orientation on the cyclo\u00adpenta\u00adnone ring. In agreement with an earlier proposed reaction mechanism, the amide group and the methyl group adjacent to the chloro substituent adopt equatorial positions and relative cis orientation, whereas the Cl substituent itself and the methyl group adjacent to the amide have axial orientations relative to the mean plane of the five-membered ring. The conformation of the mol\u00adecule is stabilized by one classical N\u2014H\u22efO (2.18\u2005\u00c5) and one non-classical C\u2014H\u22efO (2.23\u2005\u00c5) hydrogen bond, each possessing an S(6) graph-set motif. The crystal packing is defined by several non-classical intra\u00admolecular hydrogen bonds, as well as by partial stacking of the aromatic rings.The structure of the title compound, C Compounds (3) and (4) show similar 1H and 13C NMR spectra, making the direct assignment of the relative orientation of the cyclo\u00adpenta\u00adnone substituents almost impossible. The crystal structure of (1), as well as the crystal structure of another aromatic amide, cis-3-chloro-N--1,3-dimethyl-2-oxo\u00adcyclo\u00adpenta\u00adnecarboxamide, solved and reported earlier -1,3-dimethyl-2-oxo\u00adcyclo\u00adpenta\u00adne\u00adcarboxamide with cis orientation of two methyl groups, see Fischer et al. (1985Dimer (2) forms in the oxa-Diels\u2013Alder reaction of two methacryloyl chloride mol\u00adecules and, in the presence of a Lewis acid \u2005\u00c5 for N1. The conformation of the amide is stabilized by one classical N1\u2014H1\u22efO1 (2.18\u2005\u00c5) and one non-classical C10\u2014H10\u22efO2 (2.23\u2005\u00c5) hydrogen bonds graph-set motif \u2005\u00c5], the aromatic rings of neighboring mol\u00adecules show partial stacking with several short contacts centered near their nitro-substituent: C14\u22efC13i , and C13\u22efN2i [3.1860\u2005(14)\u2005\u00c5]. The C7\u2014H7A\u22efO1i hydrogen bond (2.53\u2005\u00c5) provides additional cohesion between neighboring enanti\u00adomeric mol\u00adecules in the columns \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01], and along the a axis by C7\u2014H7C\u22efO4ii non-classical hydrogen bonds \u2212x\u00a0+\u00a01, y\u00a0+\u00a0z\u00a0+\u00a0The crystal packing is governed by several short contacts, which may be classified as non-classical hydrogen bonds and readily afforded large transparent X-ray quality crystals upon slow evaporation of CHCl3/heptane solution (m.p. 402\u2013403\u2005K). 1H NMR : \u03b4 8.89 , 8.26\u20138.16 , 7.78\u20137.70 , 2.91\u20132.78 , 2.49\u20132.40 , 2.12\u20132.05 , 2.05\u20131.98 , 1.75 , 1.51 . 13C NMR : \u03b4 212.4, 168.9, 143.7, 143.3, 125.0, 119.3, 69.7, 55.0, 35.6, 29.4, 25.0, 24.1. MS (EI): m/z (%) 310\u2005(85) [M]+., 173\u2005(85) [M\u2013NHAr]+. HRMS (EI): m/z [M]+ calculated for C14H15ClN2O4 310.07203, found 310.07170.The title compound was prepared as described by Warneke al. 2014 by reactUiso(H) values were fixed at 1.5Ueq(C) for methyl H atoms, and 1.2Ueq for all other carrier atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S1600536814017711/hg5403sup1.cifCrystal structure: contains datablock(s) 1. DOI: 10.1107/S1600536814017711/hg54031sup2.hklStructure factors: contains datablock(s) 1. DOI: 1017486CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The three mol\u00adecular components are held together through hydrogen bonds.The crystal structure of the title salt hydrate contains discrete DABCOH 6H13N2+\u00b7S2O32\u2212\u00b72H2O, contains a centrosymmetric cyclic motif of eight hydrogen-bonded mol\u00adecular subunits. Two DABCOH+ cations are linked to two water mol\u00adecules and two thio\u00adsulfate anions via O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds, respectively. Two other water mol\u00adecules close the cyclic motif through O\u2014H\u22efO contacts to the first two water mol\u00adecules and to the two thio\u00adsulfate anions. A second pair of DABCOH+ cations is N\u2014H\u22efO hydrogen bonded to the two anions and is pendant to the ring. Adjacent cyclic motifs are bridged into a block-like arrangement extending along [100] through O\u2014H\u22efO inter\u00adactions involving the second pair of water mol\u00adecules and neighbouring thio\u00adsulfate anions.The crystal structure of the hydrated title salt, 2C The thio\u00adsulfate anion exhibits approximate Cv3 symmetry. However, in the crystal it has C1 symmetry with S\u2014O distances in the range 1.4688\u2005(8) to 1.4898\u2005(8)\u2005\u00c5 and an S\u2014S bond length of 2.0047\u2005(4)\u2005\u00c5, and O\u2014S\u2014O and S\u2014S\u2014O angles ranging from 107.47\u2005(4) to 110.48\u2005(5)\u00b0. In both DABCOH+ cations, the three N\u2014C bonds involving the protonated N atom are elongated [mean value 1.499\u2005(2)\u2005\u00c5] compared to the three N\u2014C bonds involving the non-protonated N atoms [mean value 1.472\u2005(4)\u2005\u00c5].The asymmetric unit Fig.\u00a01 consistsvia charge-assisted N\u2014H\u22efO hydrogen bonds to two DABCOH+ cations. The third oxygen atom (O2) of the anion acts as a hydrogen-bond acceptor for one of the water mol\u00adecules (O4). The second hydrogen bond involving this water mol\u00adecule is directed towards a symmetry-related thio\u00adsulfate anion. The second water mol\u00adecule (O5) is the donor of one O\u2014H\u22efO hydrogen bond to the other water mol\u00adecule and of one N\u2014H\u22efO hydrogen bond to one of the DABCOH+ cations. Numerical details of the hydrogen-bonding inter\u00adactions are given in Table\u00a01+ cations global, I. DOI: 10.1107/S2056989016001535/wm5264Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016001535/wm5264Isup3.cmlSupporting information file. DOI: 1449673CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Weak \u2018secondary\u2019 Te bonds and O\u2014H\u22efO hydrogen-bonding inter\u00adactions, involving water mol\u00adecules and layer O atoms, link the layers and inter\u00adlayer species. IR spectroscopic data are also presented.A new hydrated yttrium copper tellurite nitrate, yttrium(III) copper(II) bis\u00ad[trioxidotellurate(IV)] nitrate trihydrate, has been synthesized hydro\u00adthermally in a Teflon-lined autoclave and structurally determined using synchrotron radiation. The new phase is the first example containing yttrium, copper and tellurium in one structure. Its crystal structure is unique, with relatively strongly bound layers extending parallel to (020), defined by YO Tellurite and nitrate anions (involving atoms O1\u2013O9) are clearly distinguished from water mol\u00adecules OW1\u2013OW3 by their bond-valence sums (Table\u00a012\u2212 and 2 \u00d7 H2O at 2.290\u2005(3)\u20132.497\u2005(3)\u2005\u00c5. Cu is in square-planar coordination, with four close oxygen neighbours at 1.904\u2005(3)\u20131.999\u2005(3)\u2005\u00c5. Two more oxygen ligands at 2.811\u2005(4) and 2.817\u2005(4)\u2005\u00c5 complete an octa\u00adhedron that is very elongated due to the Jahn-Teller distortion. Te1 is trigonal\u2013pyramidally coordin\u00adated by three oxygen atoms at 1.883\u2005(3)\u20131.911\u2005(3)\u2005\u00c5. Three \u2018secondary bonds\u2019 to O atoms at 2.657\u2005(3)\u20132.837\u2005(3)\u2005\u00c5 complete a polyhedron that can be described as an octa\u00adhedron that is very distorted due to the lone-pair stereoactivity. Te2 has very similar coordination, with three primary Te\u2014O bonds of 1.893\u2005(3)\u20131.905\u2005(3)\u2005\u00c5 and three secondary bonds of 2.681\u2005(4)\u20132.798\u2005(3)\u2005\u00c5. In each case, two of the secondary bonds provide additional bracing within the {Y\u22efCu\u22efTe} layer, while the third is to a nitrate oxygen , and thus provides weak bridging between the layers and inter\u00adlayer species. The nitrate oxygen atom O9 makes a seventh very distant ligand for both Te1 [3.231\u2005(4)\u2005\u00c5] and Te2 [3.350\u2005(4)\u2005\u00c5], further than the shortest Te\u22efCu distances and with bond valences < 0.05 valence units, using the parameters of Mills & Christy are defined by YOs Table\u00a01. Within isty 2013.et al. 4]4\u2212 of {Cu\u22efTe\u22efCu\u22efTe} squares running parallel to [001] 8 polyhedra 2] and its Er\u2014Cl and Er\u2014Br analogues , DTGS (Deuterated Triglycine Sulfate) detector, 4\u2005cm al. 2013. Numeric3)2(NO3)(H2O)3 were synthesized hydro\u00adthermally. For the synthesis, Y(NO3)3\u00b76H2O , Cu(NO3)2\u00b73H2O (Sigma\u2013Aldrich \u226599%) and Te 200\u2005mm mesh were used as starting materials. A 1:1:1 molar ratio of the reagents in 20\u2005ml water was reacted in a Teflon autoclave bomb at 473\u2005K for 3 days. Crystals of YCu(TeO3)2(NO3)(H2O)3 were separated manually from a blue powder of undetermined composition in a few percent yield. Several unsuccessful attempts were made to synthesize YCu(TeO3)2(NO3)(H2O)3 from a stoichiometric mixture of the reagents, using the molar ratio 1:1:2. We also were unsuccessful in producing new compounds, with the same structure type or not, using La, Ce, Nd or Gd in place of Y.Dark blue prisms of YCu\u2005\u00c5.Single crystal X-ray diffraction experiments were carried out on the micro-focus macromolecular beam line MX2 of the Australian Synchrotron. Details of data collection and structure refinement are provided in Table\u00a0410.1107/S2056989016011464/wm5307sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016011464/wm5307Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016011464/wm5307sup3.pdfSupporting information file. DOI: 1493330CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecules form a closely spaced lattice structure in which neighbouring porphyrins are oriented in inversion-related dimers.The title compound contains one independent mol\u00adecule which exhibits an overall 48H60N4O2)], contains one independent mol\u00adecule. The average Ni\u2014N bond length is 1.917\u2005(13)\u2005\u00c5. The mol\u00adecules are arranged in a closely spaced lattice structure in which neighbouring porphyrins are oriented in inversion-related dimers. The nickel(II) porphyrin is characterized by a significant degree of a ruffled (Bu1) conformation with small contributions from saddle (Bu2) and wave (y) [gE(y)], as determined using normal structural decomposition. Disorder in the 2,5-di\u00ad\u00admeth\u00adoxy\u00adphenyl substituent was modelled over two positions with a 60% occupancy for the major moiety. One of the ethyl groups is also disordered over two positions and was modelled with the major moiety being present in 51.3% occupancy.The asymmetric unit of the title nickel(II) porphyrin, [Ni(C There are many studies available on metal coordination porphyrinato]nickel(II), which has an average Ni\u2014N bond length of 1.925\u2005\u00c5 and the meso carbon (Cm) can be used to determine structural differences between similar porphyrins and differences within the individual porphyin structure. The C\u03b1\u2014Cm(but\u00adyl)\u2014C\u03b1 angle of 119.12\u2005(2)\u00b0 is smaller than the C\u03b1\u2014Cm(H)\u2014C\u03b1 angle, and the C\u03b1\u2014Cm\u2014C\u03b1 angle at 123.2\u2005(2)\u00b0 is similar to both C\u03b1\u2014Cm(H)\u2014C\u03b1 angles, 122.1\u2005(3)\u00b0 (C20) and 124.8\u2005(3)\u00b0 (C15). The 2,5-di\u00admeth\u00adoxy\u00adphenyl group is tilted at an angle of 75.80\u2005(7)\u00b0 from the 24-atom least-squares plane of the porphyrin ring.The average Ni\u2014N distance is 1.917\u2005(13)\u2005\u00c5. The largest deviation occurs at the Ni\u2014N2 bond [1.906\u2005(2)\u2005\u00c5], which lies between both substituted et al., 1998ruffled (Bu1) with small contributions from saddle (Bu2) and wave (y) [gE(y)] , [gE(x)] and domed (Au2), which is similar to both highly substituted and other Ni(II) porphyrins . The tilt of the pyrrole rings against the 24-atom plane are N1 [24.85\u2005(8)\u00b0], N2 [25.22\u2005(8)\u00b0], N3 [15.79\u2005(10)\u00b0] and N4 [17.58\u2005(8)\u00b0], with the highest deviation from the mean plane associated with the pyrrole rings closest to the butyl group at C5. The maximum deviations from the least-squares plane are associated with the meso C atoms. C5 deviates from the least-squares plane by 0.880\u2005(2)\u2005\u00c5, whereas C10, C15 and C20 deviate from the plane at 0.551\u2005(2), 0.512\u2005(3) and 0.667\u2005(2)\u2005\u00c5, respectively. Table\u00a01A conformational analysis was performed using the NSD method developed by Shelnutt and co-workers (Shelnutt )] Fig.\u00a02. There an-butyl group (C51 > C54) with the phenyl meth\u00adoxy unit, specifically between H54A\u22efC104, at 2.851\u2005(4)\u2005\u00c5, the meth\u00adoxy group (O1 > C111) with the ethyl group (C181 > C182) between O1\u22efH18C at 2.552\u2005(4)\u2005\u00c5, the meth\u00adoxy group (O2B > C108) with the ethyl group (C21 > C22) between O2B\u22efH22A at 2.486\u2005(3)\u2005\u00c5 and the ethyl group (C121 > C122) with the C15 atom, between C15\u22efH12E at 2.833\u2005(3)\u2005\u00c5. However, there are no \u03c0\u2013\u03c0 inter\u00adactions or hydrogen bonds evident in the crystal structure.The unit cell of the title compound consists of two mol\u00adecules, each at a distance of 4.949\u2005\u00c5 from the 24-atom mean plane of the other. The mol\u00adecules are arranged in a closely spaced lattice structure in which ethyl groups and butyl groups point towards each other to form a cage-like inversion-related dimer Fig.\u00a03. Mol\u00adecuet al. (1992a\u2014Cm(H)\u2014Ca angles (122.85\u2013123.58\u00b0) compared to the title compound. We also determined the structure of nickel(II) with and without deuterated chloro\u00adform nickel(II) and zinc(II) derivative exhibits angles that are similar to the title compound, 122.12\u2013122.35\u00b0 for the substituted meso-positions and 123.42\u2013123.78\u00b0 for the unsubstituted meso-positions. The average Ni\u2014N bond length of 1.923\u2005\u00c5 is comparable to that of the title compound. The zinc derivative of this compound exhibits a larger average metal\u2013nitro\u00adgen bond length of 2.054\u2005\u00c5 and wider C\u03b1\u2014Cm\u2014C\u03b1 angles, 124.85\u2013125.95\u00b0 for the substituted meso-positions and 126.81\u2013127.78\u00b0 for unsubstituted meso-positions, as to be expected for zinc porphyrins.A search of the Cambridge Structural Database . The solution was heated to room temperature and over the course of 1\u2005h added to a solution of nickel(II) yielding purple crystals of the title compound . The compound was recrystallized from a solution of 1%vol MeOH in CH2Cl2 layered with hexane to yield single crystals suitable for X-ray diffraction.The title compound was prepared as reported previously (Senge Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms. Disorder in the 2,5-di\u00admeth\u00adoxy\u00adphenyl substituent was modelled over two positions with a 60% occupancy for the major moiety. The ethyl group at C12 was modelled over two positions with the major moiety being present in 51.3% occupancy. Restraints and constraints were used to model the disorder with SHELXL2014 I, publication_text. DOI: 10.1107/S2056989015020058/wm5222Isup2.hklStructure factors: contains datablock(s) I. DOI: 1427139CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Each Cd2+ cation is coordinated by two N atoms and one O atom of the tridentate ligand and three chloride anions, forming a distorted CdNOCl3 octahedron. Each pair of adjacent metal cations is linked by two bridging chloride ligands, resulting in a dinuclear complex unit. The mol\u00adecular conformation is stabilized by intra\u00admolecular N\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds. In the crystal, mol\u00adecules are linked by nonclassical C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds into a three-dimensional network. In addition, \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.777\u2005(2) and 3.631\u2005(2)\u2005\u00c5] contribute to the stabilization of the crystal packing.The title compound, [Cd DOI: 10.1107/S1600536814010630/rz5124Isup2.hklStructure factors: contains datablock(s) I. DOI: 1001915CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S)-ABOC], two amino acid residues [(S)-Ala and (S)-Phe], and protecting groups of Boc and OBn. The tetra\u00admer folds into a right-handed mixed 11/9 helix stabilized by intra\u00admolecular i,i + 3 and i,i-1 C=O\u22efH\u2014N hydrogen bonds. The oligomers are linked into chains by N\u2014H\u22efO=C hydrogen bonds with the chloro\u00adform solvent mol\u00adecules inter\u00adcalated between the folded chains via C\u2014H\u22efO=C inter\u00adactions.In the title compound, the \u03b1,\u03b2-hybrid peptide contains two non-proteinogenic amino acid residues [( S)-2-[(S)-(1-{2-[(S)-(1-{[(tert-but\u00adoxy)carbon\u00adyl]amino}\u00adbicyclo\u00ad[2.2.2]octan-2-yl)formamido]\u00adpropanamido}\u00adbicyclo\u00ad[2.2.2]octan-2-yl)formamido]-3-phenyl\u00adpropano\u00adate chloro\u00adform monosolvate, C42H56N4O7\u00b7CHCl3, the \u03b1,\u03b2-hybrid peptide contains two non-proteinogenic amino acid residues of (S)-1-amino\u00adbicyclo\u00ad[2.2.2]octane-2-carb\u00adoxy\u00adlic acid [(S)-ABOC], two amino acid residues of (S)-2-amino\u00adpropanoic acid [(S)-Ala] and (S)-2-amino-3-phenyl\u00adpropanoic acid [(S)-Phe], and protecting groups of tert-but\u00adoxy\u00adcarbonyl (Boc) and benzyl ester (OBn). The tetra\u00admer folds into a right-handed mixed 11/9 helix stabilized by intra\u00admolecular i,i\u00a0+\u00a03 and i,i\u00a0\u2212\u00a01 C=O\u22efH\u2014N hydrogen bonds. In the crystal, the oligomers are linked by N\u2014H\u22efO=C hydrogen bonds into chains along the a-axis direction. The chloro\u00adform solvent mol\u00adecules are inter\u00adcalated between the folded chains via C\u2014H\u22efO=C inter\u00adactions.In the title compound, phenyl ( S)-1-amino\u00adbicyclo\u00ad[2.2.2]octane-2-carb\u00adoxy\u00adlic acid [(S)-ABOC] is a \u03b22,3,3-tris\u00adubstituted bicyclic amino acid which exhibits a high propensity to induce both a reverse turn into short peptides and helices in oligoureas and in \u03b1,\u03b2-hybrid peptides and the NH of the \u03b1-residue (i\u00a0+\u00a03) and two C9 pseudocycles between the CO of the \u03b1-residue (i) and the NH of the \u03b2-residue (i\u00a0\u2212\u00a01). The backbone torsion angles are quite similar to those of the characteristic 11/9 helix reported in the same \u03b1,\u03b2-hybrid oligomers \u2005\u00c5] and a C\u2014H\u22efO hydrogen bond [C\u22efO = 3.071\u2005(4)\u2005\u00c5].The inter\u00admolecular inter\u00adaction N2\u2014H2\u22efO5i Table\u00a01 connectson Fig.\u00a02. In the iPrCO instead of Boc are not isomorphous. This latter crystallized in the space group P21 with two independent mol\u00adecules in the asymmetric unit. One independent mol\u00adecule shows a single fully folded 11/9 helix as the title compound while the hydrogen-bond network is incomplete in the other mol\u00adecule. The last C9 hydrogen bond between the carbonyl of the Phe residue and the \u03b2-residue amide proton was disrupted by the incorporation of a water mol\u00adecule parameters were fixed at 1.2Ueq for methine, methyl\u00adene, aromatic groups and NH groups, and at 1.5Ueq(C) for methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015016941/is5415sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015016941/is5415Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015016941/is5415Isup3.cmlSupporting information file. DOI: 1423394CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal packing features C\u2014H\u22ef\u03c0 and \u03c0\u2014\u03c0 inter\u00adactions, which generate a three-dimensional network.In the title compound, C 25H20ClN3O, the C=N bond of the central imine group adopts an E conformation. The mean planes through the essentially planar carbazole [maximum deviation = 0.052\u2005(2)\u00c5] and quinoline [maximum deviation = 0.050\u2005(2)\u2005\u00c5] ring systems form a dihedral angle of 50.2\u2005(1)\u00b0. In the crystal, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 and \u03c0\u2014\u03c0 inter\u00adactions [centroid\u2013centroid distances ranging from 3.635\u2005(2) to 3.739\u2005(2)\u2005\u00c5], forming a three-dimensional supra\u00admolecular network.In the title compound, C The sum of the bond angles around N1 (360.05\u00b0) of the pyrrole ring is in accordance with sp2 hybridization. Atom Cl1 deviates from the plane of the attached quinoline ring system by 0.100\u2005(1)\u2005\u00c5. The geometric parameters of the title mol\u00adecule agree well with those reported for similar structures , 3.739\u2005(2), 3.739\u2005(2), 3.635\u2005(2) and 3.635\u2005(2)\u2005\u00c5, respectively, forming a three-dimensional supra\u00admolecular network \u2212x, \u2212y, 1\u00a0\u2212\u00a0z; (ii) 1\u00a0\u2212\u00a0x, y, z; (iii) 1\u00a0\u2212\u00a0x, \u2212y, 1\u00a0\u2212\u00a0z and (iv) \u2212x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z).In the crystal, mol\u00adecules are linked by two C\u2014H\u22ef\u03c0 inter\u00adactions; the first one between the benzene H atom of the carbazole ring system and the benzene ring of an adjacent mol\u00adecule, with a C1\u2014H1\u22efH-carbazol-3-amine (1\u2005mmol) and sulfated SnO2-fly ash catalyst (50\u2005mg) in water (15\u2005ml) and was refluxed at 353\u2005K for 5\u201310 minutes. The completion of the reaction was monitored by TLC (ethyl acetate and hexane as an eluent 20%). After completion, the reaction mixture was cooled to ambient temperature. Then di\u00adchloro\u00admethane (20\u2005ml) was added and the organic layer filtered, dried on anhydrous Na2SO4 and the solvent removed using a rotary evaporator. The crude product was purified by column chromatography on silica gel (200 mesh) with hexane and ethyl acetate (4:1) as eluent to afford the title compound in good yield (10%). Red blocks suitable for X-ray diffraction analysis were obtained by recrystallization from di\u00adchloro\u00admethane solution at room temperature.A 25\u2005ml round-bottom flask was charged with dimedone (1\u2005mmol), 2-chloro-6-meth\u00adoxy\u00adquinoline-3-carbaldehyde (1\u2005mmol) 9-ethyl-9Uiso(H)=1.5Ueq for methyl H atoms and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989015005794/hb7384sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989015005794/hb7384Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015005794/hb7384Isup3.cmlSupporting information file. DOI: 1027676CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two independent cations are conformationally similar with the comparative dihedral angles between the central pyridine ring and the three benzene substituent rings being 3.0\u2005(2), 36.4\u2005(1) and 24.2\u2005(1)\u00b0, and 3.7\u2005(2), 36.5\u2005(1) and 24.8\u2005(1)\u00b0, respectively. In the crystal, the cations, anions and water mol\u00adecules are linked through O\u2014H\u22efO and O\u2014H\u22efBr hydrogen bonds, forming an insular unit. Within the cations there are also intra\u00admolecular N\u2014H\u22efO hydrogen bonds. Adjacent centrosymmetrically related aggregates are linked by \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine ring and a benzene ring in both cations [ring-centroid separations = 3.525\u2005(3) and 3.668\u2005(3)\u2005\u00c5], forming chains extending across the ac diagonal. Voids between these chains are filled by dichloromethane molecules.The asymmetric unit in the structure of the title compound, C DOI: 10.1107/S2056989015021386/zs2352Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015021386/zs2352Isup3.molSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015021386/zs2352Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015021386/zs2352fig1.tif. DOI: The asymmetric unit in the structure of the title compound, with displacement ellipsoids shown at the 50% probability level. Hydrogen bonds are shown as dashed lines.Click here for additional data file.10.1107/S2056989015021386/zs2352fig2.tifx y z . DOI: x\u00a0+\u00a0y\u00a0+\u00a0z operation.The result of superposition of one independent cation with another shifted by an Click here for additional data file.10.1107/S2056989015021386/zs2352fig3.tifA x y z . DOI: A indicates the symmetry operator \u2212x\u00a0+\u00a01, y\u00a0\u2212\u00a0z\u00a0+\u00a0Insular hydrogen bonded aggregates in the structure. Hydrogen bonds are shown as dashed lines. Suffix Click here for additional data file.10.1107/S2056989015021386/zs2352fig4.tif. DOI: Chains formed by \u03c0\u2013\u03c0 stacking inter\u00adactions between aromatic ring systems in adjacent H-bonded frameworks.1436252CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mononuclear complex exhibits a distorted tetra\u00adhedral coordination geometry about the metal atom, arising from one S atom of the benzene\u00adcarbo\u00adthio\u00adamide ligand, two P atoms of two tri\u00adphenyl\u00adphosphane mol\u00adecules and one bromide ion. An intra\u00admolecular N\u2014H\u22efBr hydrogen bond is observed and in the crystal structure, inversion dimers linked by pairs of N\u2014H\u22efBr and C\u2014H\u22efBr hydrogen bonds are observed. In addition, C\u2014H\u22ef\u03c0 inter\u00adactions occur, leading to [101] chains. 7H7NS)(C18H15P)2], was obtained from the reaction of silver(I) bromide with benzene\u00adcarbo\u00adthio\u00adamide (C7H7NS) and tri\u00adphenyl\u00adphosphane (C18H15P) in the mixed solvent of aceto\u00adnitrile and ethanol. The mononuclear complex exhibits a distorted tetra\u00adhedral coordination geometry about the metal atom, arising from one S atom of a benzene\u00adcarbo\u00adthio\u00adamide ligand, two P atoms of two tri\u00adphenyl\u00adphosphane mol\u00adecules and one bromide ion. An intra\u00admolecular N\u2014H\u22efBr hydrogen bond is observed and in the crystal structure, inversion dimers linked by pairs of N\u2014H\u22efBr and C\u2014H\u22efBr hydrogen bonds are observed. In addition, C\u2014H\u22ef\u03c0 inter\u00adactions occur, leading to [101] chains. Hirshfeld-surface analyses are presented and discussed.The title complex, [AgBr(C I-containing phospho\u00adrus and sulfur donor ligands have been studied and published extensively in recent years (C18H15P)2] previously synthesized by us (C18H15P)2], 2.5580\u2005(5)\u2005\u00c5. The Ag\u2014P bond lengths of 2.4682\u2005(7) and 2.4671\u2005(6)\u2005\u00c5 for Ag1\u2014P1 and Ag1\u2014P2, respectively, are similar to those of the Ag\u2014P bond lengths in [AgCl(C7H7NS)(C18H15P)2] [2.4529\u2005(5) and 2.4578\u2005(5)\u2005\u00c5], and similar to the Ag\u2014P distances of analogous tetra\u00adhedrally coordinated AgI complexes such as [Ag(NO3)(C2H3N3S)(C18H15P)2] \u00b72NO3 and C17\u2014H17\u22efBr1i [3.789\u2005(3)\u2005\u00c5] hydrogen bonds between a pair of adjacent complex mol\u00adecules; these are similar to the those in the above-mentioned complex [AgCl(C7H7NS)(C18H15P)2] \u2005\u00c5].In the crystal, the dimeric inter\u00admolecular inter\u00adactions are generated through a crystallographic inversion center by linking through the N1\u2014H1et al., 2004Crystal Explorer 3.1 and outside (de) of the Hirshfeld surface analyse the inter\u00admolecular inter\u00adactions via the mapping of normd, as depicted in Fig.\u00a04a\u2013d) for Hirshfeld surfaces of the complex are shown with characteristic pseudo-symmetry wings in the upper left and lower right sides of the de and di diagonal axes that represent the overall 2D fingerprint plot and those delineated into H\u22efH, H\u22efBr/Br\u22efH, and C\u22efH/H\u22efC contacts are shown in Fig.\u00a05a\u2013d, respectively. The fingerprint plot of H\u22efH contacts represented by the largest contribution within the Hirshfeld surfaces (60.8%) are shown as one distinct pattern with a minimum value of de + id \u223c2.6\u2005\u00c5. The reciprocal H\u22efBr/Br\u22efH contacts consist of 5.4% of the total Hirshfeld surface with de + id \u223c3.3\u2005\u00c5, exhibited by two symmetrical narrow pointed wings indicating the inter\u00admolecular hydrogen-bond inter\u00adactions N1\u2014H1A\u22efBr1 and C17\u2014H17\u22efBr1 in the crystal packing. The presence of C\u2014H\u22ef\u03c0 inter\u00adactions on the fingerprint plot, which contribute 29.7% of overall Hirshfeld surface, are indicated by de + id \u223c3.0\u2005\u00c5.For the title complex, the Hirshfeld-surfaces analysis bromide was dissolved in the mixed solvent of 15\u2005ml of aceto\u00adnitrile and 15\u2005ml of ethanol and then tri\u00adphenyl\u00adphosphane was added. The mixture was refluxed for 2\u2005h at 343\u2005K and a white precipitate was formed. After that, benzene\u00adcarbo\u00adthio\u00adamide was added and continually refluxed for 5\u2005h. At that time, the white precipitate dissolved. The clear yellow solution was filtered and left to evaporate at room temperature. After a day, pale-yellow blocks of the title compound were filtered off and dried Uiso(H) = 1.2 Ueq(C). N-bound H atoms were found from difference maps and refined isotropically with distance restraint N\u2014H = 0.85\u20130.86\u2005\u00c5.Crystal data and details of structure determination are summarized in Table\u00a0210.1107/S2056989016009518/hb7592sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016009518/hb7592Isup2.hklStructure factors: contains datablock(s) I. DOI: 1484796CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The PH 21H21O5PS\u00b7H2O\u00b7CH2Cl2, the phospho\u00adnium\u2013sulfonate zwitterion has the acidic H atom located on the P atom rather than the sulfonate group. The S\u2014O bond lengths [1.4453\u2005(15)\u20131.4521\u2005(14)\u2005\u00c5] are essentially equal. In the crystal, the water mol\u00adecules bridge two zwitterions via Owater\u2014H\u22efOsulfonate hydrogen bonds into a centrosymmetric dimer. The dimers are further linked by weak CAr\u00adyl\u2014H\u22efOsulfonate hydrogen bonds into chains extending along [100]. The PH+ group is not involved in inter\u00admolecular inter\u00adactions.In the title compound, C Addition of an H+ at phospho\u00adrous should not add significant steric congestion and therefore it is not surprising that 1 also adopts the exo3 conformation. The Ometh\u00adoxy\u22efP distances, 2.7691\u2005(14) and 2.7940\u2005(14)\u2005\u00c5, are shorter than the sum of the O and P van der Waals radii (3.35\u2005\u00c5). The O3\u22efH1(P1) distance is 2.44\u2005(2)\u2005\u00c5.Compound up Fig.\u00a01. The S\u2014Oexo3 conformation versus alternative exo2 and exo1 conformations was investigated by DFT calculations using the hybrid exchange-correlation functional PBE0 or two (exo1) meth\u00adoxy groups were rotated away from the PH+ group. The exo2 and exo1 conformers were calculated to be 1.2 and 2.5 kcal mol\u22121 less stable than the exo3 isomer, respectively. The HOMO of the exo3 conformer is comprised of p orbitals of the sulfonate O atoms, while the LUMO is delocalized over the phenyl rings and P\u2014Caromatic bonds and 1.98\u2005(3)\u2005\u00c5 (H2O)][2-{(o-CF3-Ph)2P}-4-Me-benzene\u00adsulfonate] . The structure of the tri\u00adethyl\u00adammonium salt of 2-[bis\u00ad(2-meth\u00adoxy\u00adphen\u00adyl)phos\u00adphanyl]benzene\u00adsulfonate has also been reported . The product was purified by recrystallization . Crystals of 1\u00b7H2O\u00b7CH2Cl2 (I) suitable for the X-ray diffraction analysis were obtained by layering Et2O on a CH2Cl2 solution of 1 at 277\u2005K.Compound Uiso(H) set to 1.2\u20131.5Ueq(C). The P- and O-bound H atoms were located in a difference Fourier map and refined isotropically.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016000669/cv5502sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016000669/cv5502Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016000669/cv5502Isup3.cmlSupporting information file. DOI: 1447138CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Melting points of compounds Cu3V2O8 and Cu2V2O7 are 780\u00b0C and 790\u00b0C, respectively. The addition of small amounts of Mg (II), MgxCu3-xV2O8 (x\u2009<\u20091.0) and MgyCu2-yV2O7 (y\u2009<\u20090.5) fused compositions, was not sufficient to stabilize structures at 800\u00b0C. For the Mg2CuV2O8 (x\u2009=\u20092.0) composition fired at 800\u00b0C, Mg (II) incorporated into the monoclinic Cu3V2O8 structure stabilizes this crystalline phase. At 1000\u00b0C, orthorhombic Mg3V2O8 structure from this composition was obtained. Solid solutions with orthorhombic symmetry were detected from the prepared compositions fired at 1000\u00b0C when 1.0\u2009\u2264\u2009x\u2009\u2264\u20093.0. The difference of coloration of Cu, Mg vanadates might be explained by the presence of a strong charge transfer band in visible spectra.In this study, MgThe online version of this article (doi:10.1186/s40064-015-0908-8) contains supplementary material, which is available to authorized users. Charge transfer effects in which an electron is transferred between an anion and a cation are often responsible for intense colours as in, for example, permanganate (purple) and chromates (yellow). In solids, there is an additional source of colour; it involves the transition of electrons between energy bands. Colour may be measured in a quantitative way by spectroscopic techniques. At higher frequencies than the infrared, electronic transitions associated with d-level splitting, impurity ions, crystal defects, etc., are possible. Many of these occur in the visible region and are responsible for colour.3V2O8 compound with monoclinic symmetry and space group P121/c1 is similar to the orthorhombic structure of Mg3V2O8 compound with space group Cmca and MgyCu2-yV2O7 (0\u2009\u2264\u2009y\u2009\u2264\u20092) compositions was made to investigate the possible formation of solid solutions with orthovanadate or divanadate structure. Coloration of these materials was also measured and related with structure in prepared compositions. A partial substitution of Cu (II) by Mg (II) ions might increase the melting point of materials with orthovanadates and divanadates structures. These potential solid solutions could be applied in ceramic industry.In this study, structural characterization of MgxCu3-xV2O8 (0\u2009\u2264\u2009x\u2009\u2264\u20093) and MgyCu2-yV2O7 (0\u2009\u2264\u2009y\u2009\u2264\u20092) compositions were synthesized by the chemical coprecipitation method. The starting materials were Cu(NO3)2\u2009\u00b7\u20092.5H2O , MgCl2.6H2O (Panreac) of reagent grade chemical quality, and NH4VO3 . The stoichiometric amount of NH4VO3, Cu(NO3)2\u2009\u00b7\u20092.5H2O and MgCl2\u2009\u00b7\u20096H2O was added on 200\u00a0mL of water with vigorous stirring at room temperature. These starting materials were added in solid state and the concentration of the various cations is different in each prepared composition. After that, a solution of ammonium hydroxide was added dropwise until pH\u2009=\u20098. The obtained precipitates were dried by an infrared lamp and dry samples were fired at 300\u00b0C for 12\u00a0hours, 600\u00b0C for 12\u00a0hours, 800\u00b0C for 1\u00a0hour and 1000\u00b0C for 1\u00a0hour.MgxCu3-xV2O8 (0\u2009\u2264\u2009x\u2009\u2264\u20093) and MgyCu2-yV2O7 (0\u2009\u2264\u2009y\u2009\u2264\u20092) fired compositions to investigate the possibility of formation of solid solution in these synthetic conditions. The diffraction patterns were collected running from 5 to 110 o2\u03b8, using monochromatic CuK\u03b1 radiation, a step size of 0.02 o2\u03b8 and a sampling time of 10\u00a0s. The initial structural information was obtained of the Inorganic Crystal Structure Database Rietveld . Unit ceUV\u2013vis-NIR spectroscopy (diffuse reflectance) allows the Cu (II) site and the V (V)-O and Cu (II)-O charge bands in samples to be studied. A Jasco V-670 spectrophotometer was used to obtain the UV\u2013vis-NIR (ultraviolet visible near infrared) spectra in the 200 to 2500\u00a0nm range. X-Rite spectrophotometer was used to obtain CIEL*a*b* colour parameters on fired samples: L* is the lightness axis (black (0)\u2009\u2192\u2009white (100)), a* the green (\u2212)\u2009\u2192\u2009red (+) axis, and b* is the blue (\u2212)\u2009\u2192\u2009yellow (+) axis CIE .xCu3-xV2O8 (0\u2009\u2264\u2009x\u2009\u2264\u20093) and MgyCu2-yV2O7 (0\u2009\u2264\u2009y\u2009\u2264\u20092) compositions. XRD patterns from MgxCu3-xV2O8 and MgyCu2-yV2O7 compositions are shown in Additional files Tables\u00a0xCu3-xV2O8 (0\u2009\u2264\u2009x\u2009<\u20091.0) compositions the major crystalline phase is the triclinic Cu3V2O8 polymorph at 600\u00b0C. Cu3V2O8 compound melts incongruently at 780\u00b0C. This fact can explain the melting of samples with a presence higher than 50% in Cu3V2O8 triclinic crystalline phase . This crystalline phase with triclinic structure is not detected in samples fired at 800 and 1000\u00b0C.In Mg3V2O8 structure and monoclinic symmetry was obtained at 300 and 600\u00b0C in compositions with 1.0\u2009\u2264\u2009x\u2009\u2264\u20092.0. This crystalline phase with monoclinic Cu3V2O8 structure is also present when 1.5\u2009\u2264\u2009x\u2009\u2264\u20092.0 at 800\u00b0C. The crystalline phase with the structure of MgCu2V2O8 compound (\u03b2\u2009=\u200990.44 (2) and Z\u2009=\u20094) is detected when 1.0\u2009\u2264\u2009x\u2009\u2264\u20092.0 at 800\u00b0C but the crystalline phase with the ordered metal distributions in Mg2CuV2O8 compound (\u03b2\u2009=\u2009116.22 (3), Z\u2009=\u20092, Cu1 in 2a sites and Mg1 in 4e sites, ICSD-404851) is not detected in conditions of this study. In the prepared Mg2CuV2O8 composition (MgxCu3-xV2O8 with x =2.0), crystalline phase with Cu3V2O8 structure and monoclinic symmetry in 2b sites and M2 in 4e sites) is the only crystalline phase detected at 800\u00b0C. Figure\u00a02V2O8 (x\u2009=\u20091.0), Mg2CuV2O8 (x\u2009=\u20092.0) and Mg2.5Cu0.5V2O8 (x\u2009=\u20092.5) compositions fired at 800\u00b0C.A crystalline phase with Cu3V2O8 structure is obtained when 2.0\u2009\u2264\u2009x\u2009\u2264\u20093.0 at 600\u00b0C and when 2.5\u2009\u2264\u2009x\u2009\u2264\u20093.0 at 800\u00b0C. At 1000\u00b0C, this phase is the major crystalline phase in the unfused samples (x\u2009\u2265\u20091.0). Figure\u00a0xCu3-xV2O8 compositions with x\u2009=\u20091.0, x\u2009=\u20092.0 and x\u2009=\u20092.5 fired at 1000\u00b0C. The orthorhombic Mg3V2O8 crystalline phase is the major phase in them.Crystalline phase with orthorhombic MgCu divanadates and Mg divanadates are present together with Cu and Mg orthovanadates in some samples.2V2O7 structure (orthorhombic symmetry) is only present in MgyCu2-yV2O7 compositions when y\u2009<\u20090.5 at 600\u00b0C. These compositions melt at 800\u00b0C. Diffraction peak intensity associated with this crystalline phase is weak or very weak at 300\u00b0C.Crystalline phase with \u03b1-Cu2V2O7 structure (monoclinic symmetry) is more extended than the crystalline phase with \u03b1-Cu2V2O7 structure in the prepared compositions. At 300\u00b0C, crystalline phase with \u03b2-Cu2V2O7 structure is present when y\u2009<\u20090.5 with diffraction peaks of medium intensity. This crystalline phase is present when 0.25\u2009\u2264\u2009y\u2009<\u20091.50 at 600\u00b0C and when 0.50\u2009\u2264\u2009y\u2009\u2264\u20090.75 with diffraction peak of strong or medium intensity. Figure\u00a00.25Cu1.75V2O7 composition fired at 600\u00b0C.Presence of crystalline phase with \u03b2-Cu2V2O7 structure (triclinic symmetry) is present in compositions when 0.25\u2009\u2264\u2009y\u2009\u2264\u20090.50 at 300\u00b0C but its intensity of diffraction peak is weak at 600 and 800\u00b0C is detected in a compositional range more extended than the crystalline phase with triclinic Mg2V2O7 structure from MgyCu2-yV2O7 prepared compositions. Crystalline phase with monoclinic \u03b1-Mg2V2O7 structure is identified when 0.75\u2009\u2264\u2009y\u2009\u2264\u20092.00 at 600\u00b0C, when 0.50\u2009\u2264\u2009y\u2009\u2264\u20092.00 at 800\u00b0C, and when 1.50\u2009\u2264\u2009y\u2009\u2264\u20092.00 at 1000\u00b0C.Crystalline phase with \u03b1-Mg2V2O7 structure is observed when 1.5\u2009\u2264\u2009y\u2009\u2264\u20092.00 in compositions fired at 800\u00b0C and only when y\u2009=\u20092.0 at 300\u00b0C.Crystalline phase with triclinic MgTables\u00a0xCu3-xV2O8 compositions solid solutions with triclinic Cu3V2O8 structure are obtained when 0.0\u2009\u2264\u2009x\u2009\u2264\u20091.0 at 600\u00b0C. At this temperature, the c unit cell parameter increases slightly with the replacement of Cu (II) ion by a slightly smaller one (Mg (II)) from MgxCu3-xV2O8 compositions when 0.0\u2009\u2264\u2009x\u2009\u2264\u20091.0. Ionic radii values do not explain this fact. A slight contraction of unit cell is expected. Structural distortion might explain the c increase with incorporation of Mg (II) ions in a structure (Tena From Mgure Tena .xCu3-xV2O8 (x\u2009\u2265\u20091.0) compositions fired at 800\u00b0C is noticeable in monoclinic Cu3V2O8 structure (detected when 1.5\u2009\u2264\u2009x\u2009\u2264\u20092.0) and it indicates the formation of solid solutions with monoclinic Cu3V2O8 structure in 1.5\u2009\u2264\u2009x\u2009\u2264\u20092 compositional range at 600\u00b0C and 800\u00b0C.Variation of unit cell parameters obtained from Mg3V2O8 structure are also obtained when 2.5\u2009\u2264\u2009x\u2009\u2264\u20093.0 at 600, 800\u00b0C and when 1.0\u2009\u2264\u2009x\u2009\u2264\u20093.0 at 1000\u00b0C. Figure\u00a0xCu3-xV2O8 samples fired at 1000\u00b0C/1\u00a0h. At 1000\u00b0C, the a and b unit cell parameters decrease with the replacement of Cu (II) ion by a slightly smaller one (Mg (II)). Average changes of interatomic distances are very slight. Changes in intensities with composition are detected in some samples is slightly different at 600 and 800\u00b0C in the conditions of this study. This slight difference between the two crystalline phases with the same structure is evidenced with differences in unit cell parameters values obtained. In MgyCu2-yV2O7 compositions fired at 600\u00b0C, the values of a and \u03b2 parameters decrease when y increases in \u03b2-Cu2V2O7 structure . At 800\u00b0C this crystalline phase is obtained with a weight fraction\u2009>\u200935% only when 0.5\u2009\u2264\u2009y\u2009\u2264\u20090.75. In crystalline phase with \u03b1-Mg2V2O7 structure, \u03b2 angle value is close to 100\u00b0 at 300\u00b0C and it is close to 90\u00b0 at 800\u00b0C. When two crystalline phases with this structure are detected in the same sample, unit cell parameters are 0.2\u00a0\u00c5 or 8 degrees (\u03b2 parameter) longer and wider in one of them. This fact is in accordance with the formation of solid solutions in this monoclinic \u03b1-Mg2V2O7 structure with the replacement of Mg (II) ion by Cu (II) and with an important distortion structural in these formed solid solutions. Solid solutions with monoclinic \u03b1-Mg2V2O7 structure are obtained when 0.5\u2009\u2264\u2009y\u2009\u2264\u20092.0 at 600 and 800\u00b0C. At 1000\u00b0C, these last solid solutions are obtained when y\u2009\u2265\u20091.5, including all unmelted samples and Mg (II) ions are also six.Coordination number of V (V) ion in Cu and Mg orthovanadate and divanadate structures is four. Coordination number of Cu (II) ion is four and five and coordination number of Mg (II) ion is six in these structures. Values obtained from samples are in accordance with literature about these structures. In distorted monoclinic structure of CuyCu2-yV2O7 compositions considering all the crystalline phases in each composition and its weight fraction in samples fired at 600, 800 and 1000\u00b0C. These M-O interatomic distances increase with magnesium amount (y) when 0\u2009\u2264\u2009y\u2009\u2264\u20091.25. This increased average M-O distance is coincident with destabilization of structures of Cu divanadate and a change is observed when y\u2009>\u20091.25. Structures of Cu divanadate are unstable with temperature and structures of Mg divanadates are stable at 1000\u00b0C when 1.5\u2009\u2264\u2009y\u2009\u2264\u20092.0 and MgyCu2-yV2O7 (0\u2009\u2264\u2009y\u2009\u2264\u20092) compositions fired at 600, 800 and 1000\u00b0C. Visible spectra obtained from MgyCu2-yV2O7 (0\u2009\u2264\u2009y\u2009\u2264\u20092) compositions fired at 800\u00b0C/1\u00a0h and MgxCu2-xP2O7 (0\u2009\u2264\u2009x\u2009\u2264\u20091.5) compositions fired at 800\u00b0C/5d (Tena 9) a transition d-d is allowed. Experimentally electronic spectra of Cu (II) ion are often characterized by a single highly asymmetric band. In this study spectra show a strong absorbance in 700\u20131400\u00a0nm wavelength range with the absorption maximum at 800\u2013900\u00a0nm. It can be associated with Cu (II) d-d transition. Bands due to d-d transitions are not expected from V (V) ion. Strong absorbance in 430\u2013600\u00a0nm wavelength range (in visible wavelength range) with the absorption maximum depending of copper amount in the sample is detected in MgxCu3-xV2O8 (0\u2009\u2264\u2009x\u2009\u2264\u20093) and MgyCu2-yV2O7 (0\u2009\u2264\u2009y\u2009\u2264\u20092) compositions at 300\u00b0C/12\u00a0h, 600\u00b0C/12\u00a0h, 800\u00b0C/1\u00a0h and 1000\u00b0C/1\u00a0h. The strong absorbance in 350\u2013600\u00a0nm wavelength range is not observed in MgxCu2-xP2O7 compositions and MgyCu2-yV2O7 (y\u2009=\u20092) compositions. It is associated with charge transfer between Cu-O in orthovanadates and divanadates when 0\u2009\u2264\u2009x\u2009<\u20093 or when 0\u2009\u2264\u2009y\u2009<\u20092. When x\u2009=\u20093 or y\u2009=\u20092, the charge transfer is associated with V-O charge transfer (\u03bb\u2009<\u2009430\u00a0nm in studied temperature range). Maximum of this band is observed at higher wavelength when copper amount is hight than when the copper amount is low. This strong absorbance in visible wavelength range explains the colour of these materials. At 600 and 800\u00b0C, coloration from samples with vanadate structures is red-brown, orange and yellow and is very different to the weak blue coloration obtained from samples with phosphate structures (Tena xCu3-xV2O8 (x\u2009\u2260\u20093) compositions and in MgxCu2-yV2O7 (y\u2009\u2260\u20092) compositions with different crystalline phases detected by XRD and MgyCu2-yV2O7 (0\u2009\u2264\u2009y\u2009\u2264\u20092) samples fired at 600, 800 and 1000\u00b0C are shown in Table\u00a0CIE L*, a* and b* parameters of Mg3V2O8 structure is detected. The best yellow colour is obtained from Mg2CuV2O8 (x\u2009=\u20092.0) solid solution with monoclinic Cu3V2O8 structure. This is the only crystalline phase detected at 800\u00b0C in conditions of this study. The Mg (II) incorporated into this structure stabilizes this crystalline phase at temperatures higher than 780\u00b0C (melting point of Cu3V2O8). In Mg2CuV2O8 composition fired at 800\u00b0C, average distances of M1-O \u2009=\u20091.9587\u00a0\u00c5 and M2-O \u2009=\u20092.1129\u00a0\u00c5 are obtained. Yellow colorations are not obtained from Cu, Mg divanadates.Interesting yellow colorations are obtained when x\u2009=\u20091.5 and 2.0 at 600\u00b0C/12 and when x\u2009=\u20092.0 at 800\u00b0C/1\u00a0h. In these samples that development yellow colorations, crystalline phase with monoclinic Cu0.5Cu1.5V2O7 (y\u2009=\u20090.5) fired at 600\u00b0C /12\u00a0h is the most noticeable colour but it is unstable at 800\u00b0C /1\u00a0h. Dark brown colour is obtained from this composition at 800\u00b0C. The major crystalline phase detected by XRD is the crystalline phase with \u03b2-Cu2V2O7 structure in this sample fired at 600\u00b0C.The colour red-brown obtained from Mg2V2O7 and Cu3V2O8 or \u03b2-Cu2V2O7 and \u03b1-Mg2V2O7 structures is detected by XRD.Orange colour is obtained when y\u2009=\u20090.75, 1.00 and 1.25 at 600\u00b0C/12\u00a0h and when y\u2009=\u20091.25 and 1.50 at 800\u00b0C/1\u00a0h. In these orange materials, a mixture of monoclinic crystalline phases with \u03b2-CuyCu2-yV2O7 compositions fired at 600\u00b0C/12\u00a0h are obtained. These solid solutions with monoclinic Cu3V2O8 structure are obtained when 1.5\u2009\u2264\u2009x\u2009\u2264\u20092.0 at 600\u00b0C and 800\u00b0C. Solid solutions with orthorhombic Mg3V2O8 structure are obtained when 2.5\u2009\u2264\u2009x\u2009\u2264\u20093.0 at 600, 800\u00b0C and when 1.0\u2009\u2264\u2009x\u2009\u2264\u20093.0 at 1000\u00b0C. In this study, the most stable solid solutions are obtained with Mg orthovanadate structure (orthorhombic).From MgyCu2-yV2O7 compositions, the formation of two kind of solid solutions with \u03b2-Cu2V2O7 and \u03b1-Mg2V2O7 structures is detected. Solid solutions with monoclinic \u03b2-Cu2V2O7 structure are obtained when 0.25\u2009\u2264\u2009y\u2009\u2264\u20091.50 at 600\u00b0C and when 0.50\u2009\u2264\u2009y\u2009\u2264\u20090.75 at 800\u00b0C. Solid solutions with monoclinic \u03b1-Mg2V2O7 structure are obtained when 0.5\u2009\u2264\u2009y\u2009\u2264\u20092.0 at 600 and 800\u00b0C and showing an important structural distortion. At 1000\u00b0C, solid solutions with \u03b1-Mg2V2O7 structure are obtained in 1.5\u2009\u2264\u2009y\u2009\u2264\u20092.0 compositional range. It is proposed the existence of a new polymorph of Mg2V2O7 compound with \u03b1-Mn2V2O7 structure detected when y\u2009\u2265\u20091.25 at 300\u00b0C.From MgxCu3-xV2O8 (0\u2009\u2264\u2009x\u2009<\u20093) and MgyCu2-yV2O7 (0\u2009\u2264\u2009y\u2009<\u20092) compositions which is associated with charge transfer between Cu-O in orthovanadates and divanadates. Wavelength of the abrupt change in absorbance is in accordance with Cu-O interatomic distances in these structures. In this study, the best yellow colour is obtained from Mg2CuV2O8 (x\u2009=\u20092.0) solid solution with monoclinic Cu3V2O8 structure. The colour red-brown is obtained from Mg0.5Cu1.5V2O7 (y\u2009=\u20090.5) fired at 600\u00b0C /12\u00a0h and it is unstable at 800\u00b0C /1\u00a0h. This red-brown colour is obtained when the average M-O distances are the shortest from divanadates. Orange colour is also obtained from some divanadates when average M-O distance is long.Strong absorbance in visible spectra is detected in MgStructural changes must be also considered to explain the colour of these materials. Thus, yellow colorations are obtained from orthovanadates and red-brown and orange colorations are obtained from divanadates."} +{"text": "The cationic diruthenium tetra\u00adacetate core lies on a crystallographic inversion center with Ru\u2014Ru and Ru\u2014N bond lengths of 2.2738\u2005(3) and 2.2920\u2005(17)\u2005\u00c5, respectively. The Ru\u2014Ru\u2014N bond angle is close to linear at 176.48\u2005(4)\u00b0, and a significant \u03c0-stacking inter\u00adaction of 3.5649\u2005(16)\u2005\u00c5 is seen between overlapping pyridine rings of adjacent cations.The title compound, [Ru Then, 3-chloro\u00adpyridine was added and the solution allowed to stir for 5\u2005min at room temperature. The volume of the solution was then reduced to 5\u2005ml under vacuum and allowed to cool to 278\u2005K overnight. The crystalline product was collected via suction filtration. Yield = 0.098\u2005g (63%). Crystals suitable for X-ray diffraction were obtained by slow diffusion of diethyl ether into a 1,2-di\u00adchloro\u00adethane solution of the complex. IR (cm\u22121): 2947 (\u03bdC\u2014H), 1447 (asym. \u03bdCOO), 1396 (sym. \u03bdCOO), 841, (\u03bdPF6), 766 (\u03bdC\u2014Cl), 692 (\u03b4C\u2014CH3). UV\u2013vis ): 427 (2.95), 263 (4.05), 210 (4.33).Synthesis of the title compound followed an earlier method developed in our lab (Vamvounis F2(obs) and F2, PLATON I. DOI: 10.1107/S2414314622002498/wm4161Isup2.hklStructure factors: contains datablock(s) I. DOI: 2156199CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Calophyllum gracilentum, is reported. The mol\u00adecule is characterized by a xanthone skeleton of three fused six-membered rings plus an additional fused pyrano ring and one 3-methyl\u00adbut-2-enyl side chain.The crystal structure of brasixanthone B, a naturally occurring xanthone isolated from the stem bark of 23H22O5, isolated from Calophyllum gracilentum, is characterized by a xanthone skeleton of three fused six-membered rings plus an additional fused pyrano ring and one 3-methyl\u00adbut-2-enyl side chain. The core xanthone moiety is almost planar, with a maximum deviation 0.057\u2005(4)\u2005\u00c5 from the mean plane. In the mol\u00adecule, an intra\u00admolecular O\u2014H\u22efO hydrogen bond forms an S(6) ring motif. The crystal structure features inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions.The title compound , C Calophyllum, frequently referred as \u2018bintangor\u2019 or \u2018penaga\u2019 in Malay is a part of the Calophyllaceae family and 94.2\u2005(4)\u00b0, respectively, suggesting a synclinal conformation \u2005\u00c5, \u03b8 = 65.4\u2005(7)\u00b0 and \u03c6 = 38.2\u2005(7)\u00b0. The core xanthone moiety is almost planar, with maximum deviation of 0.057\u2005(4)\u2005\u00c5 from the mean plane for C16. The dihedral angles between xanthone rings are: 2.29\u2005(19)\u00b0 for B and C, 2.94\u2005(19)\u00b0 for B and D, and 0.75\u2005(19)\u00b0 between C and D. There are two methyl groups attached to atom C1 in ring A with C\u2014C distances of 1.488\u2005(6) and 1.483\u2005(6)\u2005\u00c5.The orientation of the 3-methyl\u00adbut-2-enyl side chain attached to ring O3\u22efO4, forms an S(6) ring motif. In the crystal, the mol\u00adecules are linked by inter\u00admolecular hydrogen bonds O5\u2014H1O5\u22efO4, C11\u2014H11A\u22efO3 and C12\u2014H12A\u22efO5 mol\u00adecules are stacked by \u03c0\u2013\u03c0 inter\u00adactions with an inter\u00adplanar spacing of 3.319\u2005(4)\u2005\u00c5 between corresponding xanthone rings.In the title compound Fig.\u00a01, an intr5 Table\u00a01, forming5 Table\u00a01. Inversicalophyllum gracilentum was ground and extracted with n-hexane, chloro\u00adform, ethyl acetate and methanol. Fractionation of the hexane extract by gravity column chromatography over (Merck Kieselgel No. 1.09385.1000) silica gel with elution of n-hexa\u00adne: ethyl acetate and ethyl acetate: methanol in a step-wise gradual increment in polarity. This produced 28 fractions, which were combined and pooled together as 10 sub-fractions based on the TLC profile. Fraction 5 was subjected to further isolation by column chromatography over Sephadex LH20 eluted with methanol and several more purification steps using radial chromatography over silica (Merck Kieselgel No. 1.07749.1000), eluting with an n-hexa\u00adne:ethyl acetate (8:2) mixture. Yellow needle-like crystals were obtained. The melting point was found to be 500\u2013502\u2005K of Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314622011981/pk4037sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622011981/pk4037Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622011981/pk4037Isup3.cmlSupporting information file. DOI: 2227328CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-1,2,3-triazol-1-yl)eth\u00adyl]phospho\u00adnate ester, which was then hydrolyzed under acidic conditions (HCl) to give the \u2018free\u2019 phospho\u00adnic acid. The use of HCl for the hydrolysis caused protonation of the triazole ring, rendering the compound cationic, charged-balanced by a Cl\u2212 anion.This new triazole-functionalized phospho\u00adnic acid, PTEPHCl, was synthesized by the \u2018click\u2019 reaction of the alkyl azide diethyl-(2-azido\u00adeth\u00adyl)phospho\u00adnate with phenyl\u00adacetyl\u00adene to give the dieth\u00adylphospho\u00adnate ester, which was then hydrolyzed under acidic conditions (HCl) to give the \u2018free\u2019 phospho\u00adnic acid. The use of HCl for the hydrolysis caused protonation of the triazole ring, rendering the compound cationic, charged-balanced by a Cl\u2212 anion. There are extensive hydrogen-bonding inter\u00adactions in the structure of PTEPHCl, involving the phospho\u00adnic acid (\u2013PO3H2) group, the triazolium ring and the Cl\u2212 anion.The new triazole-functionalized phospho\u00adnic acid 5-phenyl-3-(2-phosphono\u00adeth\u00adyl)-1,2,3-triazol-1-ium chloride, C This ester was then hydrolyzed in acidic conditions to give 5-phenyl-3-(2-phosphono\u00adeth\u00adyl)-1,2,3-triazol-1-ium chloride (PTEPHCl). In the present work, we report the crystal structure of the above-mentioned triazole-functional\u00adized phospho\u00adnic acid PTEPHCl.The reaction of an organic azide with an alkyne to give a triazole is a well-known process and P1\u2014O2 = 1.5513\u2005(16)\u2005\u00c5] and one \u2018short\u2019 P\u2014O bond [P1\u2014O3 = 1.4805\u2005(14)\u2005\u00c5]. The \u2018long\u2019 P\u2014O bonds correspond to the P\u2014O\u2014H moieties and the \u2018short\u2019 P\u2014O bond corresponds to the phosphoryl P=O moiety. All P\u2014O bond lengths have the expected values \u2005\u00c5 and O2\u22efCl1 = 2.9422\u2005(17)\u2005\u00c5. The phosphoryl P=O group forms a hydrogen bond with the N\u2014H portion of the triazolium ring [O3\u22efN3 2.610\u2005(2)\u2005\u00c5]. Finally, the benzene ring inter\u00adacts with a phospho\u00adnate oxygen through a weak C\u2014H\u22efO contact at 3.476\u2005(3)\u2005\u00c5 (C6\u22efO2).The phospho\u00adnic acid moiety forms four hydrogen-bonding inter\u00adactions Fig.\u00a02. Specifi\u03c0\u2013\u03c0 stacking inter\u00adactionsThere is only one type of very weak \u03c0\u2013\u03c0 stacking inter\u00adaction in the structure of 5-phenyl-3-(2-phosphono\u00adeth\u00adyl)-1,2,3-triazol-1-ium chloride. The centroid-to-centroid distance is 4.0423\u2005(15)\u2005\u00c5, with the rings being \u2018shifted\u2019 from one another (slippage distance between the rings: 2.222\u2005\u00c5) and parallel.Crystal packingb axis. The chloride anions form corrugated sheets [\u2018short\u2019 Cl\u22efCl distances at 4.9455\u2005(12)\u2005\u00c5 and \u2018long\u2019 Cl\u22efCl distances at 6.4564\u2005(9)\u2005\u00c5] that are parallel to the bc plane.Fig.\u00a03Reagents and materialsl-ascorbic acid were from Serva. Sodium sulfate was from Merck. Di\u00adchloro\u00admethane, tetra\u00adhydro\u00adfuran (THF), hydro\u00adchloric acid (37%) and nitric acid (70%) were from Scharlau. Ion-exchange-column deionized water was used.All starting materials were obtained from commercial sources and used without further purification. The reagents diethyl 2-bromo\u00adethyl\u00adphospho\u00adnate (97%), phenyl\u00adacetyl\u00adene (98+%), copper sulfate penta\u00adhydrate (99%), zinc nitrate hexa\u00adhydrate (98%) and ethyl\u00adenedi\u00adamine\u00adtetra\u00adacetic acid (98%) were from Alfa Aesar. Sodium azide and Synthesis of 5-phenyl-3-(2-phosphono\u00adeth\u00adyl)-1,2,3-triazol-1-ium chloride (PTEPHCl)et al., 2018l-ascorbic acid to produce dieth\u00adyl[2-eth\u00adyl]phospho\u00adnate ester. The reaction mixture was heated at 313\u2005K under vigorous stirring for 48\u2005h. After filtration, the filtrate was mixed with di\u00adchloro\u00admethane (50\u2005mL) and an aqueous solution of the Cu2+ chelant ethyl\u00adenedi\u00adamine\u00adtetra\u00adacetic acid and the mixture was stirred for \u223c1\u2005h. After extraction with di\u00adchloro\u00admethane (4 \u00d7 50\u2005mL) and evaporation, dieth\u00adyl[2-eth\u00adyl]phospho\u00adnate ester was obtained in solid form. Finally, the latter (0.5\u2005g) was hydrolyzed with 25\u2005mL of H2O and 30\u2005mL of HCl at 373\u2005K for 48\u2005h, giving 5-phenyl-3-(2-phosphono\u00adeth\u00adyl)-1,2,3-triazol-1-ium chloride in crystalline form . The crystal used for the data collection was handled under inert conditions. It was manipulated while immersed in a perfluoro\u00adpolyether protecting oil and mounted on a MiTeGen Micromount\u2122.Three distinct steps were followed for the syntheses of the ligand PTEP. The first step was the synthesis of diethyl-(2-azido\u00adeth\u00adyl)phospho\u00adnate, following a properly adapted published procedure \u03b4 8.51 , 7.93 , 7.67 , 4.82 , 2.52 . 13C NMR \u03b4 146.71, 131.27, 129.38, 128.28, 125.53, 121.87, 45.42, 30.45 . 31P NMR \u03b4 20.17.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314622001894/tx4001sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622001894/tx4001Isup4.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622001894/tx4001Isup5.molSupporting information file. DOI: Click here for additional data file.10.1107/S2414314622001894/tx4001Isup4.cmlSupporting information file. DOI: 2145106CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title salt, all O\u2014H and N\u2014H groups in the cations (metforminium and ammonium) and lattice water mol\u00adecules are donor groups for hydrogen bonds, giving a highly compact crystal structure. 4)4(C4H12N5)2[V10O28]\u00b76H2O, crystallizes with the deca\u00advanadate anion placed on an inversion centre in space group PThe title compound, (NH The HMetf+ monocation has its charge located mainly on N2. Furthermore, this cation is characterized by a dihedral angle of 54.85\u2005(5)\u00b0 between planes C2\u2013C4/N3\u2013N5 and C1/N1\u2013N3. This twisted geometry is observed in several other compounds of metforminium(1+). Indeed, metformin and its cations HMetf+ and H2Metf2+ are highly flexible entities: the twist angle for 93 structures recovered from the CSD \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01], as well as between the anion and a water mol\u00adecule \u00b78H2O and [H2Metf]2[NH4]2(V10O28)\u00b710H2O \u00b76H2O (ca. 5% yield). These yields are poorly reproducible, and no powder diffraction was performed on the solid phases obtained by fractional crystallization to check their purity. Therefore, we cannot rule out the presence of other crystallized compounds in this reaction.Good-quality single crystals of the title compound were obtained during the reaction between ammonium metavanadate I. DOI: 10.1107/S2414314621006349/im4012Isup2.hklStructure factors: contains datablock(s) I. DOI: 2090930CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the dihedral angle between the mean planes of the cyclo\u00adhexane and 1,3-dithietane rings is 9.1\u2005(3)\u00b0. A short S\u22efO contact is observed in the crystal. 8H8O2S2, contains a cyclo\u00adhexane-1,3-dione ring, which has a twist-boat conformation. The C2S2 ring is close to planar (r.m.s. deviation = 0.023\u2005\u00c5) and the dihedral angle between the mean planes of the cyclo\u00adhexane and 1,3-dithietane rings is 9.1\u2005(3)\u00b0. Short intra\u00admolecular S\u22efO contacts occur [2.719\u2005(5) and 2.740\u2005(5)\u2005\u00c5]. In the crystal, the mol\u00adecules are linked by weak C\u2014H\u22efS hydrogen bonds and short [3.165\u2005(5)\u2005\u00c5] S\u22efO contacts, forming (010) layers. The prevalence of these inter\u00adactions is illustrated by an analysis of the three-dimensional Hirshfeld surface and by two-dimensional fingerprint plots.The title compound, C Ketene di\u00adthio\u00adacetals are useful inter\u00admediates in organic synthesis and have been used for the preparation of heterocyclic compounds \u2005\u00c5 whereas the C8\u2014S1 and C8\u2014S2 bond lengths differ slightly . The mol\u00adecule has local sC symmetry with a non-crystallographic mirror plane passing through atoms C8, C1, C2 and C5. The dihedral angle between the cyclo\u00adhexane and dithietane rings is 9.1\u2005(3)\u00b0 and short intra\u00admolecular S1\u22efO2 [2.719\u2005(5)\u2005\u00c5] and S2\u22efO1 [2.740\u2005(5)\u2005\u00c5] contacts are observed ed Fig.\u00a01.3.b-axis direction. The mol\u00adecules are linked by C5\u2014H5A\u22efS2 hydrogen bonds (Table\u00a01ii contacts, forming (010) layers a. A contribution of 30.7% was found for the H\u22efO/O\u22efH inter\u00adactions, representing the largest contribution; these contacts are represented by the spikes in the top left and bottom right of Fig.\u00a04b. Inter\u00adactions of the type H\u22efH appear in the middle of the scattered points in the fingerprint plots with a pair of spikes at de + di = 2.5\u2005\u00c5 and comprise 25.9% of the entire surface ; the van der Waals radius for this inter\u00adaction is 2.4\u2005\u00c5, which means it is a weak inter\u00adaction. The S\u22efH/H\u22efS contacts , which account for 23.8% of the Hirshfeld surface, are displayed on the fingerprint plot as a pair of long spikes at de + di = 2.7\u00c5. This distance differs by 0.3\u2005\u00c5 from the sum of the van der Waals radii, which means it is the strongest inter\u00adaction present. The S\u22efC/C\u22efS and S\u22efO/O \u22efS contacts are seen as pairs of spikes at de + di = 3.2 and 3.05\u2005\u00c5, respectively. These distances are shorter than the sums of the van der Waals radii of 3.5 and 3.32\u2005\u00c5, respectively. The C\u22efO/O\u22efC inter\u00adactions make a contribution of 0.7% to the Hirshfeld surface , their inter\u00adatomic distances (de + di = 3.3\u2005\u00c5) being larger than the sum of the van der Waals radius (3.22\u2005\u00c5), so this inter\u00adaction is very weak in this structure. The fingerprint plot corresponding to C\u22efH/H\u22efC contacts shows a fin-like distribution of points with the edges at de + di = 2.8\u2005\u00c5.The nature of the inter\u00admolecular inter\u00adactions in (I)ot Fig.\u00a03 shows rece Fig.\u00a04c; the vts Fig.\u00a04d, whichce Fig.\u00a04h, theirts Fig.\u00a04e shows 5.et al., 2016et al., 2002trans-2,4-bis\u00ad(isoprop\u00adyl)-2,4-bis\u00ad[(2-methyl-1-thioxo)propyl\u00adsulfan\u00adyl]-1,3-dithietane cyclo\u00adhexane-1,3-dione -5,5-dimethyl-2- [3-(4- nitro\u00adphen\u00adyl)allyl\u00adidene]cyclo\u00adhexane-1,3-dione -2-[(E)-amino]\u00adcyclo\u00adhex\u00adyl}isoindoline-1,3-di\u00adone \u03bbmax, 335\u2005nm (\u025b 18760); IR : 1640 (C=O), 1H NMR (CDCl3) \u03b4 (ppm): 4.35 , 2.52 , 1.97 ; 13C NMR (CDCl3) \u03b4 (ppm): 197.28 (CO),189.73 (C=C\u2014S), 119.93 (C=C\u2014S), 37.31 (CH2\u2014CH2\u2014CH2), 33.39 (CH2\u2014S), 18.62 (CH2\u2014CH2\u2014CH2).Potassium carbonate in DMF (50\u2005ml) was well stirred at room temperature. To this mixture, cyclo\u00adhexane-1,3-dione (0.1\u2005mol) was added and the resultant solution stirred at room temperature for 20\u2005min. Carbon di\u00adsulfide was then added in one lot. The reaction mixture was stirred and kept for 10\u2005min at room temperature. Di\u00adiodo\u00admethane (0.12\u2005mol) was added dropwise over 20\u2005min and the reaction mixture stirred for 7\u2005h at room temperature. Ice\u2013water (500\u2005ml) was added to the reaction mass, the solid was filtered and washed with water, dried and recrystallized from ethanol solution to give (I)7.Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details for the title compound are summarized in Table\u00a0310.1107/S2056989022009872/hb8028sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022009872/hb8028Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022009872/hb8028Isup3.cmlSupporting information file. DOI: 2211891CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "One hydro\u00adnium ion is complexed with an ordered 18-crown-6 mol\u00adecule with H2OH\u22efOC distances of 1.90\u20132.19\u2005\u00c5, and another hydro\u00adnium ion with a disordered 18-crown-6 mol\u00adecule with distances of 1.85\u20132.36\u2005\u00c5.The structure of the title compound, [H 3O+\u00b7C2F6NO4S2\u2212\u00b7C12H24O6 or [H3O+\u00b7C12H24O6][N(SO2CF3)2\u2212], which is an ionic liquid with a melting point of 341\u2013343\u2005K, has been determined at 113\u2005K. The asymmetric unit consists of two crystallographically independent 18-crown-6 mol\u00adecules, two hydro\u00adnium ions and two bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfon\u00adyl)amide anions; each 18-crown-6 mol\u00adecule complexes with a hydro\u00adnium ion. In one 18-crown-6 mol\u00adecule, a part of the ring exhibits conformational disorder over two sets of sites with an occupancy ratio of 0.533\u2005(13):0.467\u2005(13). One hydro\u00adnium ion is complexed with the ordered 18-crown-6 mol\u00adecule via O\u2014H\u22efO hydrogen bonds with H2OH\u22efOC distances of 1.90\u2005(6)\u20132.19\u2005(7)\u2005\u00c5, and the other hydro\u00adnium ion with the disordered crown mol\u00adecule with distances of 1.85\u2005(6)\u20132.36\u2005(6)\u2005\u00c5. The hydro\u00adnium ions are also linked to the anions via O\u2014H\u22efF hydrogen bonds. The crystal studied was found to be a racemic twin with a component ratio of 0.55\u2005(13):0.45\u2005(13).The structure of the title compound, H The asymmetric unit contains two crystallographically independent ion-pairs \u20132.19\u2005(7)\u2005\u00c5, and the other hydro\u00adnium ion with the disordered 18-crown-6 mol\u00adecule with 1.85\u2005(6)\u20132.36\u2005(6)\u2005\u00c5 distances (Table\u00a01via O\u2014H\u22efF hydrogen bonds with H2OH\u22efF3C distances of 2.12\u2005(4)\u20132.14\u2005(6)\u2005\u00c5, while the hydro\u00adnium ion with the disordered crown exhibits a weak O\u2014H\u22efF inter\u00adaction [H\u22efF = 2.50\u2005(4)\u2005\u00c5].Hydro\u00adnium\u00b718-crown-6 bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfon\u00adyl)amide is an ionized form of ternary equimolar mixture of 18-crown-6, imide superacid and water, the molten salt of which is known as a hydro\u00adnium solvate ionic liquid (m.p. 341 \u2212343\u2005K) with very strong Br\u00f8nsted acidity were used to correct the geometry of the disordered crown ether mol\u00adecule and hydro\u00adnium ion, and the displacement parameters of the disordered crown ether mol\u00adecule.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314620001625/is4042sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314620001625/is4042Isup2.hklStructure factors: contains datablock(s) I. DOI: 1982024CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "R(8)22 geometry.Pairs of mol\u00adecules related by inversion symmetry are linked by inter\u00admolecular C\u2014H\u22efF contacts with 25H18ClF2N5S, comprises almost co-planar fluoro\u00adphenyl, methyl\u00adthia\u00adzolyl, pyrazolyl and chloro\u00adphenyl rings with the second fluoro\u00adphenyl ring almost perpendicular to this plane. One fluoro\u00adphenyl group is disordered over two components of occupancy ratio 0.767\u2005(10):0.233\u2005(10) related by a 24.2\u2005(8)\u00b0 twist. In the crystal, two mol\u00adecules related by inversion symmetry are linked by a pair of C\u2014H\u22efF contacts in an R(8)22 geometry.The mol\u00adecule of the title compound, C Fluoro\u00adphenyl group E is disordered over two components with an occupancy ratio of 0.767\u2005(10):0.233\u2005(10) and related by a twist of 24.2\u2005(8)\u00b0.The mol\u00adecule of the title compound Fig.\u00a01 includesA\u2013D are close to coplanar with twist angles A/B, B/C and C/D of 4.76\u2005(10)\u00b0, 6.51\u2005(11)\u00b0 and 10.46\u2005(11)\u00b0 respectively. Ring E is almost perpendicular to A\u2013D with a C/E twist angle of 72.66\u2005(3)\u00b0 for the major component of E.Rings R(8)22 geometry to form a dimer. The pyrazolyl and fluoro\u00adphenyl rings of neighbouring mol\u00adecules are almost parallel with a centroid-to-centroid distance of 3.6510\u2005(13)\u2005\u00c5.In the crystal structure, two mol\u00adecules related by inversion symmetry are linked by a pair of C\u2014H\u22efF contacts Table\u00a01 with R, N\u2032-(4-fluoro\u00adphen\u00adyl)-2-oxo\u00adpropane\u00adhydrazonoyl bromide , and tri\u00adethyl\u00adamine in anhydrous ethanol (20\u2005ml) was stirred for 2\u2005h under reflux. The solid obtained on cooling was collected by filtration, washed with ethanol, dried and recrystallized from di\u00admethyl\u00adformamide solution to give colourless crystals of the title compound in 86% yield , m.p. 243\u00b0C, IR : 1590 (N=N), 1625 (C=C), 1650 (C=N).A mixture of 3-(4-chloro\u00adphen\u00adyl)-5-(4-fluoro\u00adphen\u00adyl)-4,5-di\u00adhydro-1SHELXL, e.s.d. = 0.01 and 0.02\u2005\u00c5) and Uij components of disordered atoms\u2019 ADPs were restrained to be similar to each other if within 2.0\u2005\u00c5 distance . Refinement gave an occupancy ratio of 0.767\u2005(10):0.233\u2005(10) for the two components related by a twist of 24.2\u2005(8)\u00b0.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314620007002/zl4041sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314620007002/zl4041Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314620007002/zl4041Isup3.cmlSupporting information file. DOI: 2005280CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compounds show different packing motifs including chains mediated by N\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds. viz. N\u2032-[(E)-pyridin-3-yl\u00admethyl\u00adidene]thio\u00adphene-2-carbohydrazide, C11H9N3OS, (I), N\u2032-[(E)-pyridin-2-yl\u00admethyl\u00adidene]thio\u00adphene-2-carbohydrazide, C11H9N3OS, (II), N-methyl-N\u2032-[(E)-pyridin-2-yl\u00admethyl\u00adidene]thio\u00adphene-2-carbohydrazide, C12H11N3OS, (III) and N\u2032-[(E)-pyridin-2-yl\u00admethyl\u00adidene]-2-(thio\u00adphen-2-yl)ethano\u00adhydrazide, C12H11N3OS, (IV) are described. The dihedral angles between the thio\u00adphene ring and the pyridine ring are 21.4\u2005(2), 15.42\u2005(14), 4.97\u2005(8) and 83.52\u2005(13)\u00b0 for (I)\u2013(IV), respectively. The thio\u00adphene ring in (IV) is disordered over two orientations in a 0.851\u2005(2):0.149\u2005(2) ratio. Key features of the packing include N\u2014H\u22efNp (p = pyridine) hydrogen bonds in (I), which generate C(7) chains propagating in the [001] direction; N\u2014H\u22efNp links also feature in (II), but in this case they lead to C(6) [001] chains; in (IV), classical amide (C4) N\u2014H\u22efO links result in [010] chains; in every case adjacent mol\u00adecules in the chains are related by 21 screw axes. There are no classical hydrogen bonds in the extended structure of (III). Various weak C\u2014H\u22efX inter\u00adactions occur in each structure, but no aromatic \u03c0\u2013\u03c0 stacking is evident. The Hirshfeld surfaces and fingerprint plots for (I)\u2013(IV) are compared.The crystal structures of four thio\u00adphene\u2013carbohydrazide\u2013pyridine derivatives, The oxygen atom of the carbonyl group and the sulfur atom of the thio\u00adphene ring lie on the same side of the mol\u00adecule [S1\u2014C4\u2014C5\u2014O1 = \u22124.9\u2005(6)\u00b0] whereas atom N3 of the pyridine ring lies on the opposite side. The dihedral angle between the thio\u00adphene and pyridine rings is 21.4\u2005(2)\u00b0 and the largest twist in the mol\u00adecule occurs about the C6\u2014C7 bond [N2\u2014C6\u2014C7\u2014C8 = \u221211.8\u2005(7)\u00b0]. The N1\u2014N2 bond length of 1.384\u2005(5)\u2005\u00c5 in (I)et al., 2016cIn (I)E configuration and C5\u2014N1\u2014N2\u2014C6 = 173.74\u2005(19)\u00b0 but unlike (I)In (II)N-methyl\u00adated derivative of (II)E configuration and C5\u2014N1\u2014N2\u2014C7 = 179.40\u2005(12)\u00b0. As with (II)et al., 2020Compound (III)et al., 2016cE and C6 and C7 are close to anti about the N\u2014N bond [C6\u2014N1\u2014N2\u2014C7 = \u2212177.90\u2005(14)\u00b0]. The dihedral angle between the aromatic rings (major disorder conformation for the thio\u00adphene moiety) in (IV)In (IV)3.p (p = pyridine) hydrogen bonds: in the former, these links generate [001] C(7) chains 1 screw axis but here the graph-set motif is C(6). The packing for (IV)C(4) amide N\u2014H\u22efO hydrogen bonds p link arising from the N-methyl group. The structures of (I)X inter\u00adactions although these are presumably very weak, given their H\u22efX lengths.Geometrical data for the directional inter\u00admolecular inter\u00adactions in (I)\u2013(IV) are listed in Tables 1ns Fig.\u00a05, with adds Fig.\u00a07 leading t\u22ef\u03c0p = 4.046\u2005(2)\u2005\u00c5 (slippage = 1.546\u2005\u00c5) for (I)t\u22ef\u03c0p = 4.0509\u2005(12)\u2005\u00c5 (slippage = 1.929\u2005\u00c5) for (II)t\u22ef\u03c0p = 4.7831\u2005(9)\u2005\u00c5 for (III)t\u22ef\u03c0p = 4.643\u2005(2)\u2005\u00c5 for (IV)The shortest aromatic ring centroid\u2013centroid separations in these structures are \u03c0CrystalExplorer show the expected red spots (close contacts) in the vicinities of the various donor and acceptor atoms.In order to gain more insight into these different packing motifs, the Hirshfeld surfaces and fingerprint plots for (I)\u2013(IV) were calculated using al. 2019. The HirThe fingerprint plots for (I)\u2013(IV) decomposed into the different percentage contact types Table\u00a05 show tha4.et al., 20162\u2014C(=O)\u2014NH\u2014N=CH\u2014Q sequence. None of these structures features a pyridine ring in the \u2018Q\u2019 position.A survey of the Cambridge Structural Database l. 2016a and Card al. 2014, respect6.SHELXL were used for the Uij values of equivalent atom pairs and a SAME card was used to restrain the nearest-neighbour and next-nearest-neighbour bond distances in the two disorder components to be equal with standard deviations of 0.02 and 0.04\u2005\u00c5, respectively. The N-bound H atoms in (I)Uiso(H) = 1.2Ueq(N). All C-bound H atoms were located geometrically (C\u2014H = 0.95\u20130.99\u2005\u00c5) and refined as riding atoms with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C). The methyl group in (III)Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989022005151/zl5030sup1.cifCrystal structure: contains datablock(s) I, II, III, IV, global. DOI: 10.1107/S2056989022005151/zl5030Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989022005151/zl5030IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989022005151/zl5030IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 10.1107/S2056989022005151/zl5030IVsup5.hklStructure factors: contains datablock(s) IV. DOI: Click here for additional data file.10.1107/S2056989022005151/zl5030Isup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022005151/zl5030IIsup7.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022005151/zl5030IIIsup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022005151/zl5030IVsup9.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022005151/zl5030sup10.docxHirshfeld surfaces. DOI: 2172437, 2172436, 2172435, 2172434CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound is an di\u00adiodo Schiff base mol\u00adecule under investigation for possible anti\u00admicrobial activity as well as for a ligand for medicinal activity. 20H14I2N2O2, a di\u00adiodo-Schiff base, crystallizes in space group Pbca with one mol\u00adecule per asymmetric unit. The mol\u00adecular structure reveals two intra\u00admolecular O\u2014H\u22efN hydrogen bonds that give the mol\u00adecule a twisted structure with non-coplanar rings. In the crystal structure, the mol\u00adecular packing is stabilized by \u03c0\u2013\u03c0 stacking, hydrogen- and halogen-bonding inter\u00adactions.The title compound, C E,1\u2032E)-bis\u00ad(methanylyl\u00adidene)}bis\u00ad(4-iodo\u00adphen\u00adol) (I) covalently bound through the imine C atoms, C7 and C14 respectively, to I1\u2014Ar(O1\u2014H1) and I2\u2014Ar(O2\u2014H2) rings , and 39.37\u2005(5)\u00b0 versus I2\u2014Ar(O2\u2014H2)] \u00adphen\u00adyl]salicylaldimine to I2\u2014Ar(O2\u2014H2) ring inter\u00adactions to T repeating pattern with the I1 and I2 atoms of the I1\u2014Ar(O1\u2014H1) and I2\u2014Ar(O2\u2014H2) rings inter\u00addigitated along the c-axis direction consists of a central ring (C8\u2013C13) with gs Fig.\u00a01. Two intgs Fig.\u00a01, O1\u2014H1\u22efN)] Fig.\u00a01. The intns Fig.\u00a02. This rens Fig.\u00a02. The addns Fig.\u00a02. Along ton Fig.\u00a03.o-phenyl\u00adenedi\u00adamine , and the reaction mixture brought to reflux with vigorous stirring for 2\u2005h. Upon cooling, the title compound (I) precipitated as an orange solid, and was filtered, washed with ethanol and dried under vacuum. Crystals of (I) suitable for single-crystal X-ray diffraction were grown from acetone layered with hexane. Yield: 4.37\u2005g (83%), m.p. 212\u2013214\u00b0C. 1H NMR \u03b4 8.54 , 7.73\u20137.54 , 7.37 , 7.22 , 6.84 . +: 568.9; observed: 568.8.To a solution of 2-hy\u00addroxy-5-iodo\u00adbenzaldehyde in ethanol (200\u2005ml) was added an ethanol (10\u2005ml) solution of Crystal data, data collection and structure refinement details of (I) are summarized in Table\u00a0210.1107/S2414314622008951/gg4010sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622008951/gg4010Isup2.hklStructure factors: contains datablock(s) I. DOI: 2205660CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "It crystallizes in the monoclinic space group P21/n.A dual emissive fluorescent substituted benzocarbazole moieties and carbon\u00adyl\u2013carbonyl inter\u00adactions between the two acetyl groups.The crystal of the title compound, C We obtained N-acetyl carbazole 2 in qu\u00adanti\u00adtative yield utilizing Buchwalds\u2019 method by treatment of di\u00adphenyl\u00adphenanthrene 1 as a substrate in the presence of Pd(OAc)2 (10\u2005mol %), NaOAc (1.0 equiv.), Cu(OAc)2 (2.0 equiv.) and powdered mol\u00adecular sieves in toluene under oxygen at 393\u2005K for 24\u2005h. Single crystals of 2 were grown from the a mixture of hexa\u00adnes and DCM (v/v = 1:1) at room temperature by slow thermal evaporation.In order to obtain the benzocarbazole, which are 22.2\u2005(1), 25.7\u2005(2)\u00b0 and 50.8\u2005(2), 59.7\u2005(2)\u00b0, respectively.In the structure of 3.2 to 4.553\u2005(1)\u2005\u00c5 , acet\u00adyl\u2013acetyl dipolar inter\u00adactions of 3.459\u2005(3) to 3.689\u2005(3)\u2005\u00c5 , C\u2014H\u22ef\u03c0 inter\u00adactions of 2.935\u2005(2) to 3.314\u2005(3)\u2005\u00c5 , and \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances of 3.801\u2005(2) to 5.672\u2005(2)\u2005\u00c5 between phenyl alkynyl moieties. Specifically, the crystal is stabilized by the phenyl groups of the alkynyl moiety, which inter\u00adacts weakly with each other through \u03c0\u2013\u03c0 stacking. Furthermore, the phenyl group also inter\u00adacts with another neighboring phenyl moiety and with the phenyl alkynyl moiety through C\u2014H\u22ef\u03c0 inter\u00adactions benzo[def]carbazoledi\u00adchloro\u00admethane solvate , Pd(OAc)2 , Cu(OAc)2 , NaOAc and powdered mol\u00adecular sieves were added under air and covered with a septum. The tube was evacuated and refilled with N2. Under a positive N2 pressure, toluene (2\u2005mL) was added via a syringe followed by degassing under a weak vacuum to this tube, and it was refilled with O2 three times. The reaction mixture was sealed and stirred at 293\u2005K for 24\u2005h under an O2 atmosphere. After completion of the reaction, the solution was cooled to room temperature and diluted with ethyl acetate followed by filtration through a thin pad of Celite. The crude product was purified by flash chromatography (hexa\u00adnes/EtOAc) on silica gel to afford N-acetyl benzo[def]carbazole 2. Crystals of the title compound were obtained by thermal evaporation of the pure compound from a 1:1 solution of di\u00adchloro\u00admethane and hexa\u00adnes.To a dried reaction tube, phenanthrene 6.Uiso(H) = 1.2 or 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022004509/ex2054sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022004509/ex2054Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022004509/ex2054Isup3.cmlSupporting information file. DOI: 2101657CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Z)-ethene-1,2-bis\u00ad(di\u00adphenyl\u00adphosphine) as well as its complex with PtII are described here. The structure of the phosphine sulfide features intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions and C\u2014H\u22efS hydrogen bonds, as well as inter\u00admolecular \u03c0\u2013\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions. The structure of the platinum(II) complex features inter\u00admolecular C\u2014H\u22efCl and C\u2014H\u22efS hydrogen bonds.The crystal structures of the di\u00adsulfide derivative of ( Z)-bis\u00ad(di\u00adphenyl\u00adphosphine sulfide), C26H22P2S2 (I), along with its complex with PtII dichloride, di\u00adchloridoplatinum(II), [PtCl2(C26H22P2S2)] (II), are described here. Compound I features P=S bond lengths of 1.9571\u2005(15) and 1.9529\u2005(15)\u2005\u00c5, with a torsion angle of 166.24\u2005(7)\u00b0 between the two phosphine sulfide groups. The crystal of compound I features both intra\u00admolecular C\u2014H\u22efS hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions. Mol\u00adecules of compound I are held together with inter\u00admolecular \u03c0\u2013\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions to form chains that run parallel to the z-axis. The inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction has a H\u22efCg distance of 2.63\u2005\u00c5, a D\u22efCg distance of 3.573\u2005(5)\u2005\u00c5 and a D\u2014H\u22efCg angle of 171\u00b0 (where Cg refers to the centroid of one of the phenyl rings). These chains are linked by relatively long C\u2014H\u22efS hydrogen bonds with D\u22efA distances of 3.367\u2005(4) and 3.394\u2005(4)\u2005\u00c5 with D\u2014H\u22efA angles of 113 and 115\u00b0. Compound II features Pt\u2014Cl and Pt\u2014S bond lengths of 2.3226\u2005(19) and 2.2712\u2005(19)\u2005\u00c5, with a P=S bond length of 2.012\u2005(3)\u2005\u00c5. The PtII center adopts a square-planar geometry, with Cl\u2014Pt\u2014Cl and S\u2014Pt\u2014S bond angles of 90.34\u2005(10) and 97.19\u2005(10)\u00b0, respectively. Mol\u00adecules of compound II are linked in the crystal by inter\u00admolecular C\u2014H\u22efCl and C\u2014H\u22efS hydrogen bonds.The crystal structures of ( Unfortu2.I was solved in the ortho\u00adrhom\u00adbic space group P212121. The mol\u00adecular structure of this compound is shown in Fig.\u00a02I has P=S bond lengths of 1.9571\u2005(15) and 1.9529\u2005(15)\u2005\u00c5, P\u2014C bond lengths that range from 1.804\u2005(4) to 1.824\u2005(4)\u2005\u00c5 and a C=C bond length of 1.338\u2005(5)\u2005\u00c5. The P=S bonds are oriented in opposite directions with a S1\u2014P1\u2014P2\u2014S2 torsion angle of 166.24\u2005(7)\u00b0. The \u03c44 descriptor for fourfold coordination around both phospho\u00adrus atoms P1 and P2 is 0.94, indicating a near tetra\u00adhedral geometry of the phosphine sulfide groups and 3.360\u2005(4)\u2005\u00c5 with D\u2014H\u22efA dihedral angles of 113 and 116\u00b0, respectively (Table\u00a01D\u22efA distances of 3.367\u2005(4) and 3.394\u2005(4)\u2005\u00c5 with D\u2014H\u22efA dihedral angles of 113 and 115\u00b0, respectively. The Flack parameter for this structure is \u22120.10\u2005(5) and 97.19\u2005(10)\u00b0, respectively . The \u03c44 descriptor for fourfold coordination around the PtII center is 0.05, indicating a nearly perfect square-planar orientation of the sulfur and chlorine atoms around the metal to 1.816\u2005(9)\u2005\u00c5, with a C=C bond length of 1.312\u2005(18)\u2005\u00c5. The \u03c44 descriptor for fourfold coordination of the phospho\u00adrus atom P1 is 0.91, indicating a slightly distorted tetra\u00adhedral geometry of the groups bonded to this atom, and that this tetra\u00adhedron is more distorted than what was observed for compound I.For the Pt3.I are held together in the crystal by inter\u00admolecular \u03c0\u2013\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions \u2005\u00c5 and a D\u2014H\u22efCg angle of 171\u00b0 (Cg is the centroid of the C15\u2013C20 ring). Together, these inter\u00admolecular \u03c0\u2013\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions link the mol\u00adecules into chains that propagate parallel to the z-axis and 3.561\u2005(5)\u2005\u00c5 with D\u2014H\u22efA angles of 155 and 135\u00b0, respectively. These hydrogen-bonding inter\u00adactions occur between the supra\u00admolecular chains of compound I.Mol\u00adecules of compound s Table\u00a01. Ring C9is Fig.\u00a05. Two potII are held together by C\u2014H\u22efCl \u2005\u00c5 with a D\u2014H\u22efA angle of 141\u00b0 . Sulfur atom S1 hosts the other inter\u00admolecular hydrogen bond with atom C3(H3) (symmetry code: x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a0D\u22efA distance of 3.538\u2005(9) with a D\u2014H\u22efA angle of 133\u00b0. The inter\u00admolecular C\u2014H\u22efCl inter\u00adactions form chains of compound II that run parallel to the z-axis. These chains are then linked into a three-dimensional network through the inter\u00admolecular C\u2014H\u22efS hydrogen bonds.Mol\u00adecules of compound 4.et al., 2016I resulted in 17 hits. The majority of these hits were metal\u2013ligand complexes, where the ligand was a triazole ring bearing two di\u00adphenyl\u00adphosphine sulfide groups. Crystal structures of this ligand bonded to copper(II), zinc(II), palladium(II), and cadmium(II) were reported and elemental sulfur were combined in a round-bottom flask and dissolved in tetra\u00adhydro\u00adfuran (5\u2005mL). The reaction mixture was stirred for three\u2005h at room temperature. The solvent was removed under reduced pressure to give a white, gelatinous solid. The crude product was recrystallized from benzene (5\u2005mL) at 333\u2005K and isolated by vacuum filtration with a Hirsch funnel to give a white solid. Analysis of the solid by 31P NMR (CDCl3) showed that the target compound I was present along with trans-dppeS2 and unreacted starting material. Single crystals of compound I grew serendipitously upon slow evaporation of this solution. 31P NMR : Compound I: 32.3 ppm; trans-dppeS2: 36.6 ppm; cis-dppe: \u221222 ppm.Compound II: Equimolar amounts of compound I and Pt(PhCN)2Cl2 were combined in a small vial and dissolved in 1\u2005mL CDCl3. Crystals of compound II formed serendipitously via slow evaporation of the solvent.Compound 6.I and II, all hydrogen atoms bonded to carbon atoms were placed in calculated positions and refined as riding: C\u2014H = 0.95\u20131.00\u2005\u00c5 with Uiso(H) = 1.2Ueq(C) for vinylic and aromatic hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022011847/pk2674sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989022011847/pk2674Isup4.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989022011847/pk2674IIsup5.hklStructure factors: contains datablock(s) II. DOI: 1481532, 2226172CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Heterodimers between nearly identical mol\u00adecules are connected via three hydrogen bonds from benzylic and ester methyl\u00adene groups to phospho\u00adnate. The dimers form chains along the a-axis direction, stabilized by C\u2014H\u22efO bridges.The title compound was prepared in three steps from 12H17I2O3P, was prepared in three steps from p-xylene. Heterodimers between nearly identical mol\u00adecules are connected via three hydrogen bonds from benzylic and ester methyl\u00adene groups to phospho\u00adnate. The dimers form chains along the a-axis direction, stabilized by C\u2014H\u22efO bridges.The title compound, C The A,B dimers are connected via three slightly bent C\u2014H\u22efO hydrogen bridges: C7A\u2014H7A\u22efO9B , C14A\u2014H14B\u22efO9B , and C11B\u2014H11D\u22efO9A . Three further C\u2014H\u22efO bridges connect neighbouring dimers to form chains along the a-axis direction \u2005\u00c5 157.9\u00b0, B shifted \u22121 along a], C7B\u2014H7C\u22efO9A and C14B\u2014H14D\u22efO9A .In a project focusing on phenyl\u00adene\u00advinyl\u00adene emissive materials I, global. DOI: 10.1107/S2414314621006544/bt4116Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314621006544/bt4116Isup3.cmlSupporting information file. DOI: 2091488CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the original publication , referenFigure 1. Frequency of use of A. rhizogenes strains in the world: 1\u2014ATCC15834; 2\u2014A4+ATCC43057; 3\u2014LBA9402; 4\u2014R1000; 5\u2014K599+NCPPB2659; 6\u2014NCIB8196; 7\u2014R1601; 8\u2014C58C1; 9\u2014NCPPB1855; 10\u2014MAFF301724; 11\u2014A4T; 12\u2014TR7; 13\u2014A4RS; 14\u2014ARqual; 15\u2014TR101; 16\u2014ATCC11325; 17\u20141334; 18\u2014TR105; 19\u2014MSU440; 20\u2014MTCC532; 21\u2014HRI; 22\u2014LMG150; 23\u2014A13; 24\u2014R1600 (this is a modified figure from ref. [68]).Biomics2015, 7, 70\u2013120.Newly added reference [68] should read: Kuluev, B.R.; Vershinina, Z.R.; Knyazev, A.V.; Chemeris, D.A.; Baymiev, A.K.; Chumakov, M.I.; Baymiev, A.K.; Chemeris, A.V. Plant hairy roots are important instrumentation for researchers and powerful phytochembiofactory for manufacturers. With this correction, newly added reference [68] and the order of some references has been adjusted accordingly.The authors would like to apologize for any inconvenience caused to the readers by these changes. The original article has been updated."} +{"text": "The morpholine ring in the cation adopts a chair conformation. The structure is stabilized by C\u2013H\u22efO, O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efO hydrogen-bonding inter\u00adactions and \u03c0\u2013\u03c0 stacking. The inter\u00admolecular inter\u00adactions of the synthesized compound were qu\u00adanti\u00adfied by Hirshfeld surface analysis.The title compound was synthesized 6H15N2O+\u00b7C6H2N3O7\u2212\u00b7H2O, was synthesized via slow evaporation of an aqueous solution of picric acid with the substituted morpholine base and crystallized with one cation (C6H15N2O)+, one anion (C6H2N3O7)\u2212 and a water mol\u00adecule in the asymmetric unit. The morpholine ring in the cation adopts a chair conformation. The structure is stabilized by C\u2014H\u22efO, O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efO hydrogen-bonding inter\u00adactions and \u03c0\u2013\u03c0 stacking. The inter\u00admolecular inter\u00adactions of the synthesized compound were qu\u00adanti\u00adfied by Hirshfeld surface analysis.The title compound, C All three protons of the NH3+ group are involved in hydrogen bonding. The cation forms a strong charge-assisted hydrogen bond N5\u2014H5A\u22efO1 with the picrate anion, while H5C inter\u00adacts with O9 from the solvating water mol\u00adecule [D\u22efA = 2.741\u2005(2)\u2005\u00c5] and H5B is involved in a bifurcated hydrogen bond with O5 from a neighbouring picrate anion and O9 from other water mol\u00adecule , respectively. Additionally, the two protons of the water mol\u00adecule inter\u00adact with a picrate anion or the nitro\u00adgen atom of the morpholinyl moiety . Further geometric details of these hydrogen bonds can be found in Table\u00a01The title compound crystallizes in the triclinic up Fig.\u00a01 with twoup Fig.\u00a01. The asypara-bound nitro group is nearly coplanar with the plane of the benzene ring [dihedral angle of \u22121.0\u2005(2)\u00b0] and two ortho-oriented nitro groups are, probably as a result of repulsion with the phenolic oxygen atom, twisted from the ring plane by \u221251.9\u2005(2) and 43.8\u2005(2)\u00b0. It has been mentioned previously that the nitro groups of the picrate anion play an important role in stabilizing the crystal packing via weak coulombic inter\u00adactions , C10\u2014H10B\u22efO6 and C12\u2014H12A\u22efO2 non-classical hydrogen bonds and 4.0303\u2005(19)\u2005\u00c5, respectively, for the rings related by symmetry operations 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 2\u00a0\u2212\u00a0z and 2\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 2\u00a0\u2212\u00a0z. Finally, a centrosymmetric twelve-membered ring [(picrate)O\u2212\u22efH\u2014N\u2014H\u22efO\u2014H]2 with a third order graph set Fig.\u00a03s Table\u00a01. Severalup Fig.\u00a05. Furtheral Fig.\u00a06.CrystalExplorer 21.5 Hirshfeld surface of the title compound mapped over the limits \u22120.6471 to 1.3714 a.u. with close contacts to neighboring mol\u00adecules is shown in Fig.\u00a07et al., 2016Analysis of the Hirshfeld surface and the associated two-dimensional fingerprint plot for 2-(morpholin\u00adyl)ethyl\u00adammonium picrate monohydrate was performed with de and di represent the distances from a point on the Hirshfeld surface to the nearest atoms outside and inside the surface, respectively morpholinium] tetra\u00adkis\u00ad[(\u03bc3-phosphito)tri\u00adzinc(II)] hemihydrate morph\u00ado\u00adlinium] tetra\u00adkis\u00ad(\u03bc-iodo)\u00adtetra\u00adkis\u00ad(iodo)\u00addilead(II)] and : 3384 (O\u2014H), 2905 (NH3), 3110 (C\u2014H), 1382 (CH2), 993 (C\u2014O); 1H NMR : 8.831 , 3.63 , 3.03 , 2.58 , 2.46 . A suitable single crystal of 2-(morpholin\u00adyl)ethyl\u00adammonium picrate monohydrate was selected for X-ray diffraction studies.2-(Morpholin\u00adyl)ethyl\u00adammonium picrate monohydrate was synthesized by mixing one mole of 4-(2-ammonio\u00adeth\u00adyl)morpholine and one mole of picric acid in double-distilled water at about 303\u2005K. The solution was then allowed to evaporate at room temperature, which yielded yellow plate-like crystals of 2-(morpholin\u00adyl)ethyl\u00adammonium picrate monohydrate. The reaction scheme is shown in Fig.\u00a096.Uiso(H) = 1.2 Ueq(C). The acidic protons were localized from the residual electron-density map and refined with distance restraints (0.82\u2005\u00c5 for O\u2014H and 0.86\u2005\u00c5 for N\u2014H) and Uiso(H) = 1.2Ueq(N) and 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022011409/jq2022sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022011409/jq2022Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022011409/jq2022Isup3.cmlSupporting information file. DOI: 2222322CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S-(Pyridn-2-yl) benzo\u00adthio\u00adesters are presented with varying para-phenyl motifs . These structures presented are the first in their class. Distinct changes are observed in the inter\u00adaction types present in the crystal lattice as a direct result of the electronic influence of the para-phenyl motif.The structures of three S-(pyridin-2-yl) benzo\u00adthio\u00adesters with varying para-phenyl substituents are presented, namely, S-(pyridin-2-yl) 4-nitro\u00adbenzo\u00adthio\u00adate , S-(pyridin-2-yl) 4-methyl\u00adbenzo\u00adthio\u00adate and S-(pyridin-2-yl) 4-meth\u00adoxy\u00adbenzo\u00adthio\u00adate . This class of compounds are used in the mono-acyl\u00adation of pyrrolic species to yield multifunctional tetra\u00adpyrroles. The structures presented herein are the first of their compound class. The dominant inter\u00adactions present in this series are \u03c0\u2013\u03c0 stacking and C\u2014H\u22efO inter\u00adactions, and as the para-phenyl motif changes from electron withdrawing to electron donating , changes are observed in the inter\u00adactions present in the crystal packing, from predominant \u03c0\u2013\u03c0 stacking in 1 to exclusively C\u2014H\u22efO/N inter\u00adactions in 3.The crystal structures of three Each mol\u00adecular structure shows an S-(pyridin-2-yl) benzo\u00adthio\u00adate where the para-phenyl motif is modified, from NO2 in 1, CH3 in 2, and OCH3 in 3. All of the groups utilized herein are found extensively in the field of tetra\u00adpyrroles.The single-crystal XRD structures of title compounds 3 Figs. 2\u20134 \u25b8 \u25b8, 1\u20133, the substituted phenyl moieties are all essentially planar with the pyridine ring twisted relative to this plane. This is seen in the plane normal to plane normal angle and the torsion angle described by C8\u2014S1\u2014C6\u2014N1. The twist of the methane\u00adthio\u00adate moiety to the phenyl ring also describes the change in the angle of the rings to each other. These values are shown in Table\u00a01In all structures 1 . Furthermore, considering the respective previously determined Hammett constants, it is observed that the most electron donating is the OCH3 group in 3 (\u03c3p = \u22120.27), with 2 (CH3) lying somewhere in between (\u03c3p = \u22120.17) and 3.371\u2005(5)\u2005\u00c5]. The pyridine N1 is also an acceptor to the phenyl C12-H [D\u22efA = 3.315\u2005(5)\u2005\u00c5]. The nitro group is a dual acceptor with inter\u00adactions between O18 and one pyridyl C3-H [D\u22efA = 3.396\u2005(5)\u2005\u00c5] and also a bifurcated inter\u00adaction between O17 and phenyl C14-H and C15-H .Compound s Table\u00a02 to the c2 presents C\u2014H\u22efN-paired dimers between the H6-pyridyl protons C2-H2 and N1 and a C16-H\u22efO9 inter\u00adaction [D\u22efA = 3.460\u2005(2)\u2005\u00c5].Compound 3 presents a multitude of non-classical hydrogen-bonding inter\u00adactions, of the C\u2014H\u22efOcarbon\u00adyl and the C\u2014H\u22efNpyrid\u00adyl type (Table\u00a04D\u22efA = 3.2566\u2005(15) and 3.4270\u2005(16)\u2005\u00c5, respectively]. There is another bifurcated hydrogen-bond inter\u00adaction between the pyridine N1 and C11 and C12 , linking the mol\u00adecules head to tail. The meth\u00adoxy groups form C17-H\u22efO16 inter\u00adactions [D\u22efA = 3.4475\u2005(17)\u2005\u00c5], comprising a supra\u00admolecular synthon linking two mol\u00adecules together. The meth\u00adoxy oxygen O16 is further linked by a phenyl C14-H\u22efO16 inter\u00adaction [D\u22efA = 3.3340\u2005(15)\u2005\u00c5].Compound e Table\u00a04. The car1 and 2. Weak dimeric offset \u03c0\u2013\u03c0 stacking is observed in 1 with columns of anti-parallel non-inter\u00adacting mol\u00adecules when viewed normal to (001) x, y, \u22121\u00a0+\u00a0z; x, y, 1\u00a0+\u00a0z] is 3.850\u2005(3)\u2005\u00c5 with a slippages of 1.823 and 1.856\u2005\u00c5, respectively, and angles between planes of 0.0\u2005(2)\u00b0. In 2, \u03c0-stacking occurs only through phenyl ring pairs with the closest centroid\u2013centroid distance being 3.8783\u2005(11)\u2005\u00c5, a slippage of 1.575\u2005\u00c5, and an angle between planes of 0.03\u2005(9)\u00b0, as seen normal to the (011) plane. In 3 there is no relevant \u03c0\u2013\u03c0 stacking, with the closest centroid\u2013centroid distance being 4.0847\u2005(7)\u2005\u00c5, with a slippage of 2.042\u2005\u00c5 and an angle between the planes of 5.14\u2005(6)\u00b0.\u03c0\u2013\u03c0 stacking is evident in both 1) Fig.\u00a06. The clo4.et al., 2016S-phenyl benzo\u00adthio\u00adate . The distinct C\u2014H\u22efN inter\u00adactions seen particularly in 3 do not exist in the phenyl homologue.A search in the Cambridge Structural Database benzo\u00adthio\u00adate benzo\u00adthio\u00adate \u00adbenzene-1,4-dicarbo\u00adthio\u00adate -5.1, 2, and 3 were synthesized following the reported procedure in a solution of CH2Cl2 was added dropwise over 0.5\u2005h to a stirring solution of 2-mercapto\u00adpyridine in CH2Cl2. The solution was left to stir for a further 2\u2005h at room temperature. Throughout the addition processes, minor exotherms were noted, particularly for 1. The solution was diluted with the same volume again of CH2Cl2, and the solution was washed with NaOH (2 M), water, brine, and the organic layer then dried (MgSO4). Excess solvent was removed under reduced pressure and the title compounds were purified in the following ways: for 1, crystals were generated via hot recrystallization from ethyl acetate, and for 2 and 3, crystals were generated via precipitation from diethyl ether and hexa\u00adnes. Compound 1 was yielded in 69%, with yields for 2 and 3 comparable to those previously reported .1:Analytical data for 1H NMR \u03b4 = 8.66\u20138.68 , 8.31 , 8.14 , 7.77\u20137.81 , 7.68\u20137.70 , 7.33\u20137.37 ; 13C{1H} NMR : \u03b4 = 188.3, 150.9, 150.3, 141.3, 137.7, 130.9, 128.7, 124.3, 124.2 ppm; RF = 0.58 ; m.p. = 427\u2013429\u2005K. Multiple attempts have been made to obtain a mol\u00adecular ion peak via ESI\u2013MS and all have been unsuccessful.6.Uiso(H) = 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989023001056/yz2028sup1.cifCrystal structure: contains datablock(s) 1, 2, 3, global. DOI: 10.1107/S2056989023001056/yz20281sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989023001056/yz20282sup3.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S2056989023001056/yz20283sup4.hklStructure factors: contains datablock(s) 3. DOI: Click here for additional data file.10.1107/S2056989023001056/yz20281sup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001056/yz20282sup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001056/yz20283sup7.cmlSupporting information file. DOI: 10.1107/S2056989023001056/yz2028sup8.pdf1H, 13C, and 1H-13C-HSQC NMR spectra of compound 1, along with overlayed aromatic regions of the 1H NMR spectra of compounds 1, 2, and 3. DOI: 2239845, 2239844, 2239843CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the packing of the title compound, \u03c0\u2013\u03c0, C\u2014H\u22efO, C\u2014H\u22efBr, and N\u2014H\u22efN inter\u00adactions are present. 12H14BrN3O2, the pyrazole and benzene rings are nearly co-planar with a dihedral angle between the rings of 2.36\u2005(5)\u00b0. In the crystal, inversion dimers linked by pairwise N\u2014H\u22efN hydrogen bonds generate R22(8) loops. The dimers are linked into a three-dimensional network by weak aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid separation = 3.7394\u2005(6)\u2005\u00c5] and C\u2014H\u22efO and C\u2014H\u22efBr hydrogen bonds.In the title compound, C Very weak C2\u2014H2\u22efO2, C11\u2014H11C\u22efO2 and C12\u2014H12B\u22efO2 intra\u00admolecular inter\u00adactions are present = 3.7394\u2005(6)\u2005\u00c5, where Cg1 is the centroid of the pyrazole ring.In the extended structure, pairwise N3\u2014H3if Fig.\u00a02. The dimon Fig.\u00a03 also occ5-Bromo-1H-indazol-3-amine (1): To a solution of 5-bromo-2-fluoro benzo\u00adnitrile (1.0\u2005mmol) in ethanol (20\u2005ml) was added hydrazine hydrate (99%) (10.0\u2005mmol). The reaction mixture was heated in sealed tube at 343\u2005K for 4\u2005h and progress of the reaction was monitored by TLC. The reaction mixture was concentrated to dryness. The brown-coloured solid was purified by recrystallization from ethanol solution to afford pale-yellow needles (90%), m.p. 407\u2005K : To a solution of compound (1) (5.0\u2005mmol) in di\u00adchloro\u00admethane (40\u2005ml) was added DMAP (5.0\u2005mmol). The reaction mixture cooled to 273\u2005K and boc anhydride (5.0\u2005mmol) was added. The reaction mixture was slowly warmed to room temperature and stirred for 15\u2005h. Progress of the reaction was monitored by TLC. The reaction mixture was diluted with di\u00adchloro\u00admethane (50\u2005ml) and washed with water and brine (25\u2005ml), dried over anhydrous sodium sulfate and concentrated. The crude compound was purified by column chromatography to afford a gummy solid, which solidifies as transparent crystals after 2\u2005d (62%), m.p. 389\u2005K.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621006945/hb4388sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2414314621006945/hb4388Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314621006945/hb4388Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2414314621006945/hb4388Isup4.cmlSupporting information file. DOI: 2094667CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Water mol\u00adecules donate O\u2014H\u22efO and accept N\u2014H\u22efO hydrogen bonds, forming helical arrangements.In the crystal, mol\u00adecules of the title compound, C 24H20N2O4\u00b7H2O, crystallizes with half a mol\u00adecule of 2--N\u2032-[2-acet\u00adyl]acetohydrazide and half a water mol\u00adecule in the asymmetric unit. In the crystal, mol\u00adecules form planes parallel to (011). Two mol\u00adecules are connected by water mol\u00adecules via O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds.The title compound, C An ORTEP representation is shown in Fig.\u00a01et al., 2020The asymmetric unit comprises half a mol\u00adecule of 2--N\u2032-[2-acet\u00adyl]acetohydrazide mol\u00adecules are connected by a water mol\u00adecule via O\u2014H\u22efO and N\u2014H\u22efO bonds Fig.\u00a02. Two 2- and 2-acetohydrazide in dry ethanol (10\u2005mL) was heated with stirring under reflux for 2\u2005h. The solid formed on cooling to room temperature. It was collected by filtration, washed with ethanol, dried and recrystallized from di\u00admethyl\u00adformamide to give colourless crystals, m.p. > 300\u00b0C I. DOI: 10.1107/S241431462100314X/bt4110Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S241431462100314X/bt4110Isup3.cmlSupporting information file. DOI: 2072979CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Thereby, the titanium atom is stabilized by the Lewis bases tri\u00adphenyl\u00adphosphine oxide ( 6H11)2N]3Ti\u2014CH3 with the Lewis acid B(C6F5)3, followed by addition of the Lewis bases (C6H5)3PO and p-F\u2014C6H4CN led to the complex salts tris\u00ad(di\u00adcyclo\u00adhexyl\u00adamido)(tri\u00adphenyl\u00adphosphine oxide)titanium methyl\u00adtris(penta\u00adfluoro\u00adphen\u00adyl)borate toluene sesquisolvate, [Ti(C12H22N)3(C18H15OP)](C19H3F15B)\u00b71.5C7H8, (1), and tris\u00ad(di\u00adcyclo\u00adhexyl\u00adamido)(4-fluoro\u00adbenzo\u00adnitrile)\u00adtitanium methyl\u00adtris\u00ad(penta\u00adfluoro\u00adphen\u00adyl)borate toluene sesquisolvate, [Ti(C12H22N)3(C7H4FN)](C19H3F15B)\u00b71.5C7H8, (2), both crystallizing with 1.5 equivalents of toluene solvent mol\u00adecules. The Lewis acid\u2013base adducts (1) and (2) can be described by dative donor bonds. The packing of the complex cations, anions and solvent mol\u00adecules in the crystal structure is consolidated by an intricate three-dimensional network of non-classical C\u2014H\u22efF inter\u00adactions. Disorder of some of the cyclo\u00adhexyl groups and the toluene solvent mol\u00adecules is observed.The reaction of [(C An intricate three-dimensional network of non-classical C\u2014H\u22efF inter\u00adactions is formed in the crystals of (1) and (2), involving the C\u2014H groups of both cations and solvent mol\u00adecules as donors, and some F atoms of the [H3CB(C6F5)3]\u2212 anions as acceptors and (2) were obtained from a saturated solution of toluene at 243\u2005K.All reactions were carried out under a dry nitro\u00adgen atmosphere using Schlenk techniques or in a glove box. Solvents were dried according to standard procedures over Na/K alloy with benzo\u00adphenone as an indicator and distilled under a nitro\u00adgen atmosphere. The cationic titanium complex was synthesized by reacting tris-(di\u00adcyclo\u00adhexyl\u00adamido)\u00admethyl\u00adtitanium and tris-penta\u00adfluorphenyl\u00adborane (Adler 5.1) shows disorder of one cyclo\u00adhexyl residue (C25 to C30), located at N3, over two sets of sites, with refined site occupancy factors 0.87:0.13. Compound (2) shows disorder of two of the six cyclo\u00adhexyl residues , located at the N1 and N3 atoms, respectively, with refined site occupation factors of 0.76:0.24 and 0.85:0.15. No restraints or constraints were applied during the refinement of this kind of disorder. All non-H atoms were refined anisotropically with the exception of the minor components of the disordered parts, which have been refined isotropically. In addition, the toluene solvent mol\u00adecule in compounds (1) and (2) are disordered. In (1), one toluene mol\u00adecule (C81\u2013C87) is equally disordered over an inversion centre, and one (C74\u2013C80) over two sets of sites, with refined site occupation factors of 0.764\u2005(4):0.236\u2005(4). In (2), one toluene mol\u00adecule (C63\u2013C69) is disordered over two sets of sites, the site occupancy was constrained to 0.5 for each component. The other toluene mol\u00adecule (C70\u2013C76) is disordered over two sets of sites with refined occupation factors of 0.29\u2005(3):0.210\u2005(3), both of which are additionally disordered over an inversion centre, resulting in a disorder over four sites.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022007952/wm5649sup1.cifCrystal structure: contains datablock(s) 1, 2. DOI: 10.1107/S2056989022007952/wm56491sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989022007952/wm56492sup3.hklStructure factors: contains datablock(s) 2. DOI: 2195527, 2195526CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "HLA\u2010DRB5*01:01:01\u2010DRB1*15:01:01\u2010DQA1*01:02:01\u2010DQB1*06:02:01 haplotype.Multiple sclerosis (MS) is a chronic neurological disease believed to be caused by autoimmune pathogenesis. The aetiology is likely explained by a complex interplay between inherited and environmental factors. Genetic investigations into MS have been conducted for over 50 years, yielding\u00a0>100 associations to date. Globally, the strongest linkage is with the human leukocyte antigen (HLA) DRB3, DRB4, DRB5, DRB1, DQA1, DQB1, DPA1 and DPB1 as well as their extended haplotypes and genotypes in 100 Swedish MS patients. Results were compared to 636 population controls.Here, high\u2010resolution sequencing of HLA was used to determine the alleles of HLA\u2010DR\u2010DQ genotypes were found. Three extended HLA\u2010DR\u2010DQ genotypes were found to be correlated to MS; HLA\u2010DRB5*01:01:01\u2010DRB1*15:01:01\u2010DQA1*01:02:01\u2010DQB1*06:02:01 haplotype together withThe heterogeneity in HLA associations with MS was demonstrated; among 100 patients, 69 extended HLA\u2010DRB4*01:01:01//DRB4*01:01:01:01\u2010DRB1*07:01:01\u2010DQA1*02:01//02:01:01\u2010DQB1*02:02:01,(A) HLA\u2010DRBX*null\u2010DRB1*08:01:01\u2010DQA1*04:01:01\u2010DQB1*04:02:01, and(B) HLA\u2010DRB3*01:01:02\u2010DRB1*03:01:01\u2010DQA1*05:01:01\u2010DQB1*02:01:01.(C) HLA\u2010DRB3*01:01:02 was considered protective against MS. However, when combined with HLA\u2010DRB3*01:01:02\u2010DRB1*03:01:01\u2010DQA1*05:01:01\u2010DQB1*02:01:01, this extended haplotype was considered a predisposing risk factor. This highlights the limitations as included with investigations of single alleles relative to those of extended haplotypes/genotypes.At the allelic level, HLA\u2010DRB5*01:01:01\u2010DRB1*15:01:01\u2010DQA1*01:02:01\u2010DQB1*06:02:01.In conclusion, with 69 genotypes presented among 100 patients, high\u2010resolution sequencing was conducted to underscore the wide polymorphisms present among MS patients. Additional studies in larger cohorts will be of importance to define MS among the patient group not associated with Studies in African\u2010Americans, among whom the HLA\u2010DQB1*06:02 allele is not found on HLA\u2010DRB1*15:01 haplotypes, has attributed the risk of MS to HLA\u2010DRB1*15:01. Additional HLA\u2010DRB1\u2010DQB1 alleles and haplotypes associated with MS includeHLA\u2010DRB1*03:01\u2010DQA1*05:01\u2010DQB1*02:01 is the most common inflammatory disease affecting the central nervous system. Epidemiological findings suggest a gene\u2013environment interaction, and the pathogenesis of MS is widely assumed to be of autoimmune origin included blood samples collected from the Neurology Department of the Sk\u00e5ne University Hospital in Malm\u00f6. The MS diagnosis of the patients was validated by treating physician (Table\u00a0n\u00a0=\u00a0636) included in the HLA Next Generation Sequencing (NGS) analysis were obtained as previously described and (B) LifeGene controls .2.2DNA was extracted using Qiagen Blood Maxi Kit according to manufacturer's protocol and as previously described , and second occasion: LifeGene controls (n\u00a0=\u00a0188)]; in the meantime the resolution of typing was improved; therefore, that some alleles are written as follows in the manuscript DRB4*01:01:01//DRB4*01:01:01:01.HLA NGS was performed by Scisco Genetics . Samples were sent blindly to the investigators. HLA NGS makes use of PCR\u2010based HLA amplification and sequencing with Illumina MiSeq technology, as described in previous studies , as previously described .A reference haplotype with a similar frequency between the study groups was selected in HLA 6 haplotypes with positive (A) and 1 haplotypes with negative (B) association for MS patients in comparison to population controls were identified :HLA\u2010DRB3*02:02:01\u2010DRB1*03:01:01\u2010DQA1*05:01:01\u2010DQB1*02:01:01, (2) HLA\u2010DRB4*01:03:01\u2010DRB1*04:02:01\u2010DQA1*03:01:01\u2010DQB1*03:02:01, (3) HLA\u2010DRB4*01:03:01\u2010DRB1*04:03:01\u2010DQA1*03:01:01\u2010DQB1*03:02:01, (4) HLA\u2010DRB5*01:01:01\u2010DRB1*15:01:01\u2010DQA1*01:02:01\u2010DQB1*06:02:01, (5) HLA\u2010DRB5*02:02//DRB5*02:02:01\u2010DRB1*16:01:01\u2010DQA1*01:02:02\u2010DQB1*05:02:01 and (6) HLA\u2010DRBX*null\u2010DRB1*01:02:01\u2010DQA1*01:01:02\u2010DQB1*05:01:01.Identified haplotypes with higher frequency among MS patients compared to GP were (1) HLA\u2010DRB4*01:03:01\u2010DRB1*07:01:01\u2010DQA1*02:01//*02:01:01\u2010DQB1*03:03:02 (Table\u00a0Identified haplotypes with lower frequency among MS patients compared to GP were (1) Based on the Haplo\u2010Score, 3.3HLA\u2010DRB3, DRB4, and DRB5 alleles, 47 different HLA\u2010DRB1 alleles, 23 different HLA\u2010DQA1 alleles, 24 different HLA\u2010DQB1 alleles, 14 HLA\u2010DPA1 and 30 HLA\u2010DPB1 alleles were found. The complete results and associations for all the different alleles found in the study are reported in Supplementary Table HLA\u2010DRB3*03:01:01 , HLA\u2010DRB1*03:01:01 , HLA\u2010DQA1*03:01:01 , HLA DQB1*02:01:01 , HLA\u2010DPA1*01:03:01 and HLA\u2010DPB1*02:01:02 . Based on the Haplo\u2010Score, 9 alleles with positive and 7 alleles with negative association for MS patients in comparison to population controls were identified , HLA\u2010DRB3*01\u2010DR3\u2010DQ2, while the association with HLA\u2010DR7\u2010DQ2 appear novel.From the present study, we first report that previously found HLA\u2010associations to MS, most prominently to HLA\u2010DPB1*104:01 (predisposing), HLA\u2010DQB1*03:01 and HLA\u2010DQB1*03:03 (protective), and HLA\u2010DRB1*14:04:01 (protective) . Due to high degree of polymorphism presented, we suggest that mechanistic studies would be necessary to fully determine the role of HLA\u2010DQB1*03:01 to risk and progression of MS.Here, we could not confirm recent allelic MS associations using high\u2010resolution sequencing to HLA\u2010DRB3*01:01:02 was considered protective against MS at the allelic level, the extended genotype, abbreviated HLA\u2010DR15\u2010DQ6\u2010DRB3*01\u2010DR3\u2010DQ2, was considered a predisposing risk factor. This highlights the limitations of studies of single alleles relative to studies of extended haplotypes and genotypes. We speculate that the shifting association for HLA\u2010DRB3*01:01:02 from protective to predisposing, could be related to mechanisms of epistasis. Previously, HLA\u2010DQA1*01:02 was demonstrated to increase risk of MS if found in trans position to the haplotype of HLA\u2010DR15\u2010DQ6 and two\u2010thirds of all patients (68 of 100) encompassed haplotypes of either HLA\u2010DR15\u2010DQ6, HLA\u2010DRB3*01\u2010DR3\u2010DQ2 or HLA\u2010DRB3*02\u2010DR3\u2010DQ2.Lastly, we report heterogeneity in HLA associations to MS given that, among 100 patients, 69 different extended HLA\u2010DR15\u2010DQ6 haplotype is consistently reported, both here and elsewhere, as the major genetic contributor to MS. Some suggestions of the presence of this extended haplotype at the age at onset as compared to 6% is of interest, complementary triggers and pathways of autoimmunity could be present. It cannot be excluded that environmental factors are different between MS patients negative for HLA\u2010DR15\u2010DQ6 but positive for HLA\u2010DR7\u2010DQ2. Studies in larger and diversified cohorts would be needed to define MS in this group of patients.The group of non\u2010HLA\u2010DRB1*03:01 allele has been associated with MS. An increased risk was reported when the HLA\u2010DRB1*03:01 allele is homozygous among both patients and controls, which precluded computation of the other rare allele variants among the families that were used as templates (Lind et\u00a0al., HLA\u2010DR15\u2010DQ6 patients.A weakness of our study was the fact that the It appears overly simplistic to characterize MS as having a single aetiology and pathogenesis. The heterogeneous clinical presentation, variable clinical course, inconsistency in genetic markers, unpredictable therapeutic response and diverse histopathological findings may be indicative of divergence in the demyelination pathways (Lucchinetti et\u00a0al., HLA\u2010DR15\u2010DQ6.2.In summary, we have confirmed previous HLA associations to MS, while also offering plausible explanations to help guide future research. Using high\u2010resolution sequencing, we have highlighted the widespread polymorphisms as 69 distinct genotypes present among 100 Swedish MS patients and further studies in a larger number of patients will be needed to further delineate MS not associated with There are no conflicts of interest.Supplement MaterialClick here for additional data file.Supplement MaterialClick here for additional data file."} +{"text": "There is a high carrying rate of \u03b1\u2010thalassemia in Fujian province. However, there are few large\u2010scale studies on the correlation between genotype and phenotype in Fujian province. The purpose of this study was to analyze the phenotype and genotype in a cohort of 2923 patients with \u03b1\u2010thalassemia in Fujian province, so as to provide reference data for screening and diagnosis of \u03b1\u2010thalassemia in Fujian province.The genotype of \u03b1\u2010thalassemia was detected by PCR reverse dot blot assay, gap\u2010PCR, single PCR, nested PCR, and sequencing. Clinical and hematological indices of 2923 patients were collected, and the correlation between genotype and phenotype was analyzed.SEA/\u03b1\u03b1 was the most common genotype, accounting for 64.80%. In addition, rare \u03b1\u2010thalassemia genotypes were detected in Fujian province, including \u2010\u2010THAI/\u03b1\u03b1 (0.41%), HK\u03b1\u03b1/\u2010\u2010SEA (0.03%), and the novel \u03b1\u2010thalassemia gene mutation CD5 (GCC>ACC) (HGVS named HBA1: c.16G>A) (0.03%). Patients with deletional genotypes of \u03b1\u2010thalassemia were found to have higher RBC and lower Hb, MCV, MCH, and HbA2 than patients with non\u2010deletional genotypes of \u03b1\u2010thalassemia (p\u2009<\u20090.05).Among 10,350 patients, 2923 cases were found with \u03b1\u2010thalassemia, with a detection rate of 28.24%. Among them, \u2010\u2010The clinical phenotype of \u03b1\u2010thalassemia is influenced by molecular mechanisms. HBA1: c.16G>A mutation is a novel mutation that was first reported in Fujian province, which enriches the human hemoglobin mutation spectrum. There were double peaks at nt224, G>A, in HBA1, corresponding to CD5 (GCC>ACC). It was named HBA1: c.16G>A using the Human Genome Variation Society (HGVS) nomenclature. HBA1: c.16G>A is a novel mutation that was first reported in Fujian province. The discovery of this novel mutation in \u03b1\u2010globin gene has enriched the database of hemoglobin variants, and the detailed genetic analysis and clinical symptom description of this mutation will contribute to the further study of hemoglobin function in the future. Its pathogenesis is due to the defect of \u03b1\u2010globin gene; the synthesis of \u03b1\u2010globin peptide chain was partially or completely inhibited, resulting in hereditary hemolytic anemia. It is one of the most common monogenic diseases with the highest incidence in the world and has attracted extensive attention at home and abroad because of its fatal and disabling nature, which can lead to birth death or birth defects.However, there are few large\u2010scale studies on the correlation between genotype and phenotype in \u03b1\u2010thalassemia patients in Chinese population. In this study, 10,350 patients from Fujian province were analyzed for genotype and phenotype. Such a study may provide more data for genetic counseling and clinical diagnosis in this region.22.1This study was reviewed and approved by the Ethics Review Committee of Fujian Maternity and Child Health Hospital College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University. Signed informed consent was obtained from all participants following a detailed description of the purpose of the study. All experiments were performed in accordance with relevant guidelines and regulations.2.2From January 2019 to November 2021, peripheral blood samples of patients who underwent thalassemia\u2010related examinations in outpatients and inpatients of Fujian Maternity and Child Health Hospital College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University were collected. A total of 10,350 patients with household registration in Fujian province were screened, including 2923 cases of \u03b1\u2010thalassemia. \u03b1\u2010thalassemia was confirmed by molecular analyses. Information about patients diagnosed with \u03b1\u2010thalassemia was collected, and their genotypes and phenotypes were statistically analyzed. The median age was 27 (8.31) years. The median age was 5.5 (1.30) years for males and 28 (25.31) years for females. Exclusion criteria are as follows: (1) \u03b2\u2010thalassemia; (2) other blood diseases such as iron deficiency anemia and megaloblastic anemia; (3) malignant tumor; (4) systemic immune disorders; and (5) allergic constitution. Peripheral blood samples were collected from selected subjects and anticoagulated with EDTA\u2010K2. Approximately 2\u2009ml of the anticoagulated blood sample was used for analysis of blood cell parameters on a Sysmex XN\u20102000 automatic hematology analyzer , and the hemoglobin components and levels were analyzed using an automated capillary electrophoresis system . Serum ferritin (SF) concentration, as a measure of iron status, was determined by the chemiluminescent microparticle immunoassay (CMIA) .2.3SEA/, \u2010\u03b13.7/, and \u2010\u03b14.2/) and the mutations of \u03b1\u2010thalassemia were analyzed by PCR reverse dot blot assay using commercial kits as described. The 17 common \u03b2\u2010thalassemia was performed using PCR reverse dot blot assay with the thalassemia gene detection kit following the manufacturer's instructions.DNA was extracted using the DNA Blood Extraction Kit . The deletions (\u2010\u20102.4THAI genotype was tested using gap polymerase chain reaction (gap\u2010PCR), and HK\u03b1\u03b1 genotype was tested by single PCR and nested PCR, as described previously.\u2010\u20102.5p\u2009<\u20090.05 were considered statistically significant.All data were entered into and managed using Microsoft Excel 2007 . The data were analyzed using the Statistical Package for the Social Sciences (SPSS) version 25 . Associations between hematological indices and the variant genotypes were assessed by a nonparametric Kruskal\u2013Wallis test, presented as median . The results with 33.1Among 10,350 patients, 2923 patients with \u03b1\u2010thalassemia mutation were detected, with a detection rate of 28.24%, including 2666 cases (91.21%) of \u03b1 deletional genotype, 181 cases (6.19%) of \u03b1 mutational genotype, 12 cases (0.41%) of \u03b1 deletion combined with mutation, and 64 cases (2.19%) of \u03b1/\u03b2 complex mutants.SEA/\u03b1\u03b1 (64.80%), \u2010\u03b13.7/\u03b1\u03b1 (18.95%), \u2010\u03b14.2/\u03b1\u03b1 (4.96%), \u03b1QS\u03b1/\u03b1\u03b1 (3.63%), \u03b1CS\u03b1/\u03b1\u03b1 (1.47%), \u2010\u2010SEA/\u2010\u03b13.7 (1.20%), and \u03b1WS\u03b1/\u03b1\u03b1 (0.99%). In addition, rare \u03b1\u2010thalassemia genotypes including \u2010\u2010THAI/\u03b1\u03b1 (0.41%), HK\u03b1\u03b1/\u2010\u2010SEA (0.03%), and the novel \u03b1\u2010thalassemia gene mutation CD5 (GCC>ACC) (HGVS named HBA1: c.16G>A) (0.03%) were detected in Fujian province \u2009\u00d7\u20091012/L) (p\u2009<\u20090.05).Conversely, the levels of Hb, MCV, MCH, and HbA2 in patients with deletional genotypes of \u03b1\u2010thalassemia were lower than that in patients with non\u2010deletional genotypes of \u03b1\u2010thalassemia g/L vs 119.0 g/L, 67.5 fl vs 76.1 fl, 21.2 pg vs 24.7 pg, and 2.3 % vs 2.5 %) (p\u2009<\u20090.05). in patients with deletional genotypes of \u03b1\u2010thalassemia was higher than that in patients with non\u2010deletional genotypes of \u03b1\u2010thalassemia \u2009\u00d7\u200910SEA/\u03b1\u03b1 were found to have higher RBC, HbF, and SF and lower Hb, MCV, MCH, and HbA2 than patients with \u2010\u03b13.7/\u03b1\u03b1 (p\u2009<\u20090.05). What is more, patients with \u2010\u2010SEA/\u03b1\u03b1 were found to have higher RBC and lower Hb, MCV, MCH, and HbA2 than patients with \u2010\u03b14.2/\u03b1\u03b1 (p\u2009<\u20090.05). Age of \u2010\u2010SEA/\u03b1\u03b1 was lower than that of \u2010\u03b13.7/\u03b1\u03b1 and \u2010\u03b14.2/\u03b1\u03b1 (p\u2009<\u20090.05) Among the non\u2010deletional genotypes of \u03b1\u2010thalassemia, patients with \u03b1QS\u03b1/\u03b1\u03b1 were found to have higher RBC and HbA2 and lower MCV and MCH than patients with \u03b1CS\u03b1/\u03b1\u03b1 (p\u2009<\u20090.05). Patients with \u03b1QS\u03b1/\u03b1\u03b1 were found to have lower MCV and MCH than patients with \u03b1WS\u03b1/\u03b1\u03b1 (p\u2009<\u20090.05). In addition, patients with \u03b1CS\u03b1/\u03b1\u03b1 were found to have lower HbA2 than patients with \u03b1WS\u03b1/\u03b1\u03b1 (p\u2009<\u20090.05). that was suspected to be a \u03b1\u2010thalassemia carrier. Sequencing the full\u2010length \u03b11\u2010globin gene and of the full\u2010length \u03b12\u2010globin gene of the DNA from the peripheral blood sample was done. The results showed there were double peaks at nt224, G>A, in HBA1, corresponding to CD5 (GCC>ACC). It was named HBA1: c.16G>A using the Human Genome Variation Society (HGVS) nomenclature with\u2010\u2010SEA/\u03b1\u03b1, 554 cases (18.95%) with \u2010\u03b13.7/\u03b1\u03b1, and 145 cases (4.96%) with \u2010\u03b14.2/\u03b1\u03b1, suggesting that the most common genotype of \u03b1 \u2010thalassemia in this region was \u2010\u2010SEA/\u03b1\u03b1, which was consistent with the reports from Guangxi, Guangdong, Chongqing, and Hainan.QS\u03b1/\u03b1\u03b1 (3.63%), followed by \u03b1CS\u03b1/\u03b1\u03b1 (1.47%) and \u03b1WS\u03b1/\u03b1\u03b1 (0.99%), which were the same as those in Chongqing and slightly different from those in Guangxi, Guangdong, and Hainan,SEA/\u03b1\u03b1 was dominant in these provinces, and the frequencies were respectively 57.55%, 52.78%, and 83.87%.There are two types of \u03b1\u2010thalassemia: deletional genotype and non\u2010deletional genotype. In addition to the most common deletional genotype, rare deletional genotype of \u03b1\u2010thalassemia has been found in different populations.THAI/\u03b1\u03b1 , HK\u03b1\u03b1/\u2010\u2010SEA , and the novel \u03b1\u2010thalassemia gene mutation CD5 (GCC>ACC) in Fujian Province. \u2010\u2010THAI genotype has been gradually found in the population of southern China, mainly distributed in Guangxi, Guangdong, Taiwan, and Fujian.SEA genotype, the phenotype of \u2010\u2010THAI is manifested as microcytic hypochromic anemia and the Thai\u2010type homozygote or Thai\u2010type heterozygotes with SEA\u2010type \u03b1\u2010thalassemia are manifested as Bart's hydrops fetalis.3.7 fragment and \u03b1\u03b1\u03b1anti4.2 fragment formed by recombination and unequal transposition of the homologous X segment of the \u03b1\u2010globin gene cluster. So far, \u03b1\u03b1\u03b1anti4.2 fragments cannot be detected through the routine thalassemia diagnostic kit. The genotype of \u2010\u03b13.7/\u03b1\u03b1, HK\u03b1\u03b1/\u2010\u03b13.7, and HK\u03b1\u03b1/\u03b1\u03b1 is entirely the result of \u2010\u03b13.7/\u03b1\u03b1 by gap\u2010PCR.3.7/\u03b1\u03b1. Therefore, the symptoms of thalassemia associated with HK\u03b1\u03b1/\u03b1\u03b1 and \u2010\u2010SEA were milder than those of hemoglobin H disease associated with \u2010\u03b13.7 and \u2010\u2010SEA, and only mild globin production disorder was observed . If both husband and wife carry HK\u03b1\u03b1/\u03b1\u03b1 and \u2010\u2010SEA respectively, the fetus can be kept. However, if the HK\u03b1\u03b1/\u03b1\u03b1 or anti\u2010HK\u03b1\u03b1/\u03b1\u03b1 genotypes are incorrectly identified as \u2010\u03b13.7/\u03b1\u03b1, it may lead to incorrect clinical treatment.Three rare \u03b1\u2010thalassemia genotypes were also detected in this study, including \u2010\u2010SEA/\u03b1\u03b1/\u03b2IVS\u20102\u2010654(C\u2192T)/\u03b2N (11 cases), \u2010\u03b13.7/\u03b1\u03b1/\u03b2IVS\u20102\u2010654(C\u2192T)/\u03b2N (11 cases), \u2010\u2010SEA/\u03b1\u03b1/\u03b2CD41\u201042(\u2010CTTT)/\u03b2N (11 cases), \u2010\u03b13.7/\u03b1\u03b1/\u03b2CD41\u201042(\u2010CTTT)/\u03b2N (4 cases), and \u2010\u2010SEA/\u03b1\u03b1/\u03b2\u221228(A\u2192G)/\u03b2N (4 cases). While patients with \u03b1\u2010 and \u03b2\u2010thalassemia have milder symptoms, their offspring are more likely than the general population to develop severe thalassemia, and the long\u2010term damage is much greater. Therefore, the clinical diagnosis of concurrent \u03b1\u2010 and \u03b2\u2010thalassemia should not be ignored. According to Xiong Fu et al.,In addition, 64 cases (2.19%) with concurrent \u03b1\u2010 and \u03b2\u2010thalassemia were detected in 2923 positive samples. Patients with concurrent \u03b1\u2010 and \u03b2\u2010thalassemia have been reported to suffer from mild anemia due to a reduction in \u03b1\u2010 and \u03b2\u2010globin chain synthesis, which alleviates the imbalance caused by reduced globin chain synthesis and thus reduces the severity of anemia.SEA/\u03b1\u03b1 was lower than that of \u2010\u03b13.7/\u03b1\u03b1 and \u2010\u03b14.2/\u03b1\u03b1 (p\u2009<\u20090.05); this may be related to the number of inactivated \u03b1\u2010globin genes of \u2010\u2010SEA/\u03b1\u03b1 were more than \u2010\u03b13.7/\u03b1\u03b1 and \u2010\u03b14.2/\u03b1\u03b1. Patients with \u2010\u2010SEA/\u03b1\u03b1 were associated with more severe anemia and came to the hospital earlier for diagnosis and treatment. Chen Suqin et al.4.2 or \u2010\u03b13.7 variants. Thus, the absence of \u03b1\u2010globin peptide chains or the relative excess of \u03b2\u2010globin peptide chains is more severe in non\u2010deletional patients. The hematologic appearance of patients with non\u2010deletional genotype of \u03b1\u2010thalassemia differs from that patients with deletional genotype of \u03b1\u2010thalassemia; anemia is more severe in patients with non\u2010deletional genotype of \u03b1\u2010thalassemia.p\u2009<\u20090.05).It may be related to the damage of erythrocyte membrane caused by excess \u03b2\u2010chain oxidation and \u03b1QS\u03b1 or \u03b1CS\u03b1 chain oxidation. This results in decreased EPO production and RBC in non\u2010deletional genotypes of \u03b1\u2010thalassemia.QS\u03b1/\u03b1\u03b1's Hb was lower than \u2010\u03b13.7/\u03b1\u03b1 and \u2010\u03b14.2/\u03b1\u03b1, which was partly supported that anemia in the non\u2010deletional genotype was more severe than in the deletional genotype. Patients with \u2010\u2010SEA/\u03b1\u03b1 were found to have higher RBC, HbF, and SF, it may be related to \u2010\u2010SEA/\u03b1\u03b1 was associated with more \u03b3\u2010globin mutations, and its RBC synthesis requires more iron, so the RBC, HbF, and SF were higher than \u2010\u03b13.7/\u03b1\u03b1.SEA/\u03b1\u03b1 were lower than \u2010\u03b13.7/\u03b1\u03b1 and \u2010\u03b14.2/\u03b1\u03b1 in the three common deletional genotypes, suggesting that the severity of anemia depends on the number of inactivated \u03b1\u2010globin genes.There is significant variation in clinical severity among patients with \u03b1\u2010thalassemia, which is indirectly reflected by the span of age at first diagnosis. Age of \u2010\u2010In conclusion, the clinical manifestations and hematologic phenotypes of \u03b1 \u2010thalassemia are related to genotype. The clinical phenotype of \u03b1\u2010thalassemia is influenced by molecular mechanisms. HBA1: c.16G>A is a novel mutation that was first reported in Fujian province. The discovery of this novel mutation in \u03b1\u2010globin gene has enriched the database of hemoglobin variants, and the detailed genetic analysis and clinical symptom description of this mutation will contribute to the further study of hemoglobin function in the future.Yali Pan, Meihuan Chen, Na Lin, Liangpu Xu, and Hailong Huang designed and prepared the study. YanHong Zhang, Min Zhang, and Lingji Chen collected the literature and the data and prepared the study. All authors approved the final study.This work was funded by the National Natural Science Foundation of China (no.81970170), the National Natural Science Foundation of Fujian Province (no.2019J01510), and the Fujian provincial health technology project (no.2018\u20101\u201021).The authors confirm that they have no competing interests."} +{"text": "Erratum to:Z Gerontol Geriat 202110.1007/s00391-021-02003-5The authors would like to apologize for an error concerning the mCAM-ED and make the following corrections:Page 109, Table\u00a02, mCAM-EDColumn: Screening vs. monitoring/Items:2.\u00a0If inattention present\u202f\u2192\u2009MSQ for identifying cognitive impairment, Comprehension subtest of CTD for identifying disorganized thinkingColumn: ScoringModified CAM algorithm: 1a\u00a0(acute onset) AND 1b\u00a0(fluctuating course)\u202f+\u20092\u202f+\u2009(3\u00a0or 4) positive\u202f=\u2009diagnosed delirium1a\u00a0OR 1b\u202f+\u20092\u202f+\u2009(3\u00a0or 4) positive\u202f=\u2009suspected delirium.Page 111, Legend table\u00a02:SPMSQ: MSQ mental status questionnaire.As a\u00a0replacement for"} +{"text": "Erep) is dominant.In the crystal, hydrogen-bonding inter\u00adactions between the 2,4,6-tri\u00adamino\u00adpyrimidine cation and the nitrate anions lead to a one-dimensional supra\u00admolecular network with weak anionic inter\u00adactions forming a three-dimensional network. Energy framework analysis showed that of the components of the framework energies, electrostatic repulsion \u00b0 between the mean plane of the cation and that defined by both anions. In the crystal, hydrogen-bonding inter\u00adactions between the 2,4,6-tri\u00adamino\u00adpyrimidine cation and the nitrate anions lead to a one-dimensional supra\u00admolecular network with weak anionic inter\u00adactions forming a three-dimensional network. These inter\u00adactions were investigated using Hirshfeld surface analysis, which indicates that the most important contributions for the packing arrangement are from O\u22efH/H\u22efO (53.2%), N\u22efH/H\u22efN (12.5%) and C\u22efH/H\u22efC (9.6%) inter\u00adactions. Energy framework analysis showed that of the components of the framework energies, electrostatic repulsion (Erep) is dominant.The title compound, C The plane of the anion containing N6 is inclined to the mean plane of the cation by 3.25\u2005(6)\u00b0 while that of the other anion is inclined by 2.84\u2005(6)\u00b0. Thus the whole asymmetric unit lies close to a common plane . The ring C\u2014N bond lengths in the cation [C1\u2014N2 = 1.3531\u2005(16)\u2005\u00c5 and C2\u2014N3 = 1.3267\u2005(16)\u2005\u00c5] are only slightly altered from those in the corresponding conjugate base . Pairs of adjacent ribbons are linked by N1\u2014H1B\u22efO3, N2\u2014H2\u22efO1 and N3\u2014H3A\u22efO3 hydrogen bonds with O3\u22efCg1i = 3.1369\u2005(11)\u2005\u00c5, N6\u22efCg1i = 3.4241\u2005(12)\u2005\u00c5, N6=O3\u22efCg1i = 92.16\u2005(7)\u00b0; O5\u22efCg1ii = 3.0265\u2005(11)\u2005\u00c5; N7\u22efCg1ii = 3.5176\u2005(12)\u2005\u00c5; N7=O5\u22efCg1ii = 102.62\u2005(7)\u00b0 \u00b0 Fig.\u00a03.4.et al. 2009et al., 2007Crystal Explorer17 through white to blue . Top and bottom views of the surface together with curvedness, and shape-index plots are given in Fig.\u00a04a\u2013d. The red spots symbolize N\u2014H\u22efO contacts and C\u2014H\u22efO inter\u00adactions. The fingerprint plots , followed by N\u22efH/H\u22efN contacts at 12.5% and C\u22efH/H\u22efC contacts at 9.6% .The Hirshfeld surface is mapped over ts Fig.\u00a05 give an 2% Fig.\u00a05b, follo5% Fig.\u00a05c and C\u22ef6% Fig.\u00a05d.5.To synthesize the title compound, 20\u2005mg of 2,4,6-tri\u00adamino\u00adpyrimidine were dissolved in ethanol (10\u2005mL) and the solution stirred for 3\u2005h. A mixture of ethanol (5\u2005mL) and nitric acid (0.5\u2005mL) was taken in a separate round-bottom flask and stirred for 3\u2005h at 333\u2005K. Afterwards, the 2,4,6-tri\u00adamino\u00adpyrimidine solution was added dropwise to the above mixture. The reaction was continued for 4\u2005h at the same temperature. After completion of the reaction, a pale-yellow solution was obtained, which was filtered and kept for slow evaporation at room temperature. After 15 days, pale-yellow crystals were obtained that were suitable for data collection Fig.\u00a06.6.CrystalExplorer 17.5. Electrostatic (Eele), polarization (Epol), dispersion (Edisp), and exchange-repulsion (Erep) are the four energy variables that make up the total inter\u00admolecular inter\u00adaction energy (Etot). Cylinder-shaped energy frameworks represent the relative strengths of inter\u00adaction energies in individual directions and give the topologies of pair-wise inter\u00admolecular inter\u00adaction energies within the crystal = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022005333/mw2185sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022005333/mw2185Isup2.hklStructure factors: contains datablock(s) I. DOI: 2173730CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The layers are connected by C\u2014H\u22efCl hydrogen bonds and C\u2014H\u22ef\u03c0(ring) inter\u00adactions, forming a three-dimensional structure.The meth\u00adoxy group lies very close to the plane of the phenyl ring while the acetamido group is twisted out of this plane. In the crystal, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds form layers of mol\u00adecules parallel to the 9H10ClNO2, the meth\u00adoxy group lies very close to the plane of the phenyl ring while the acetamido group is twisted out of this plane by 28.87\u2005(5)\u00b0. In the crystal, a three-dimensional structure is generated by N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds plus C\u2014H\u22ef\u03c0(ring) inter\u00adactions. A Hirshfeld surface analysis of the inter\u00admolecular inter\u00adactions was performed and indicated that C\u22efH/H\u22efC inter\u00adactions make the largest contribution to the surface area (33.4%).In the title mol\u00adecule, C The Cl1\u2014C1\u2014C2\u2014O1 torsion angle is 52.89\u2005(12)\u00b0, illustrating a + synclinal (+ gauche) conformation about the C1\u2014C2 bond. This places atom Cl1 at 1.299\u2005(1)\u2005\u00c5 from the plane defined by C1, C2, N1 and O1.The meth\u00adoxy group lies close to the mean plane of the phenyl ring C3\u2013C8, as indicated by the C7\u2014C6\u2014O2\u2014C9 torsion angle of \u2212174.61\u2005(10)\u00b0 and atom C9 deviating by only 0.065\u2005(1)\u2005\u00c5 from the mean plane through the C3\u2013C8 ring. In contrast, the acetamido group is rotated out of the above plane with the dihedral angle between the mean plane through the C3\u2013C8 ring and that defined by N1/C2/C1/O1 being 28.87\u2005(5)\u00b0 Fig.\u00a01. The sum3.1 axes. These chains are linked by C1\u2014H1A\u22efO2 hydrogen bonds yielded 15 hits of which 13 had X = Cl and R = OEt ) (SPh)(NO2) , O\u22efH/H\u22efO (19.5%) and Cl\u22efH/H\u22efCl (20%) inter\u00adactions, respectively.The analysis was performed with 6.0.047\u2005mol of 4-methoxyaniline were dissolved in 40\u2005mL of pure acetic acid and put in an ice bath. Subsequently, chloro\u00adacetyl chloride (0.047\u2005mol) was added portionwise under stirring. At the end of the reaction, a solution of sodium acetate (35\u2005mL) was added and a solid precipitate appeared after 30\u2005min of stirring at room temperature. The resulting solid was filtered and washed with cold water, dried and recrystallized from ethanol to give the title compound as colourless crystals.\u22121) 3292 (\u03c5 N\u2014H amide), 1029 (\u03c5 N\u2014C amide), 1660 (\u03c5 C=O amide), 3073 (\u03c5 C\u2014Harom), 827 (\u03c5 C\u2014Cl), 2959 , 1H NMR (DMSO\u2013d6) \u03b4 pm: 3.74 ; 4.24 , 6.93\u20137.5 , 10.23 , 13C NMR (DMSO\u2013d6) \u03b4 ppm: 43.48 (CH2), 55.23 (CH3), 131.53 (Carom\u2014N), 155.51 (Carom\u2014O), 113.92\u2013120.92 (Carom), 164.13 (C=O); HRMS (ESI\u2013MS) (m/z) calculated for C9H10ClNO2 199.04, found 199.0105.Yield 80%, m.p. = 398.6\u2013400.3\u2005K, FT\u2013IR (1.5 for the methyl group). The N-bound H atom was found in a difference-Fourier map and refined with a DFIX 0.91 0.01 instruction and an independent isotropic displacement parameter.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698902200576X/vm2264sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S205698902200576X/vm2264Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902200576X/vm2264Isup3.cmlSupporting information file. DOI: 2175514CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The packing of the title compound features weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. 22H20O3, the dihedral angle between the aromatic rings linked by the methine group is 81.265\u2005(4)\u00b0 and the eth\u00adoxy side chain adopts an extended conformation [C\u2014O\u2014C\u2014C = 177.24\u2005(10)\u00b0]. In the crystal, weak C\u2014H\u22ef\u03c0 and C\u2014H\u22efO inter\u00adactions link the mol\u00adecules into sheets.In the title compound, C These link the mol\u00adecules into sheets lying perpendicular to the c-axis direction \u00b0. In the crystal, weak C\u2014H\u22efO Table\u00a01 and C\u2014H\u22efon Fig.\u00a02.3-Eth\u00adoxy-4-hy\u00addroxy benzaldehyde (0.20\u2005mmol) and potassium carbonate (0.40\u2005mmol) were mixed in di\u00admethyl\u00adformamide (25\u2005ml) and stirred for 0.5\u2005h. A solution of di\u00adphenyl\u00adbromo\u00admethane (0.2\u2005mmol) in ethanol (20\u2005ml) was added dropwise and the mixture was stirred at 80\u00b0C for 24\u2005h. After that, the solution was concentrated under reduced pressure. The cream precipitate of the title compound formed by adding cold water (250\u2005ml) was filtered off and washed several times with cold ethanol. Colourless slabs were recrystallized from the mixed solvents of chloro\u00adform and ethanol (1:1).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621003564/hb4378sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314621003564/hb4378Isup2.hklStructure factors: contains datablock(s) I. DOI: 2075001CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Such ICCs are of potential utility, and there arenumerous examples of phenolic compounds that are biologically active,some of which suffer from low aqueous solubility. The propensity toform ICCs sustained by the PhOH\u00b7\u00b7\u00b7PhO\u2013 supramolecular heterosynthon was evaluated through a combinationof Cambridge Structural Database (CSD) mining, structural characterizationof nine novel ICCs, and calculation of interaction energies. Our analysisof these 9 ICCs and the 41 relevant entries archived in the CSD revealedthat phenol groups can reliably form ICCs through charge-assistedPhOH\u00b7\u00b7\u00b7PhO\u2013 interactions. This conclusionis supported by hydrogen-bond strength calculations derived from CrystalExplorerthat reveal the PhOH\u00b7\u00b7\u00b7PhO\u2013 interactionto be around 3 times stronger than the phenol\u2013phenol hydrogenbond. The PhOH\u00b7\u00b7\u00b7PhO\u2013 supramolecularheterosynthon could therefore enable crystal engineering studies ofa large number of phenolic pharmaceutical and nutraceutical compoundswith their conjugate bases.Although crystalengineering strategies are generally well exploredin the context of multicomponent crystals formed by neutralcoformers , cocrystals comprised of one or moresalts are understudied. We herein addressthe design, preparation, and structural characterization of ICCs formedby phenolic moieties, a common group in natural products and drugmolecules. Organic and inorganic bases were reacted with the followingphenolic coformers: phenol, resorcinol, phloroglucinol, 4-methoxyphenol,and 4-isopropylphenol. Nine ICCs were crystallized, each of them sustainedby the phenol\u2013phenolate supramolecular heterosynthon (PhOH\u00b7\u00b7\u00b7PhO CambridgeStructural Database mining and experimental studiesof model compounds reveal that ionic cocrystals sustained by phenol\u00b7\u00b7\u00b7phenolatehydrogen bonds are formed reliably Etter systematicallyclassified common hydrogen-bonding motifs using \u201cgraph set\u201dnotation Gda(r), where G is the pattern designator that could be C (chain), R (ring),and D (dimer or other finite set); a represents the number of acceptors; d is the number of donors; and r is thenumber of atoms in the repeat unit.18 Graph settheory remains a valuable tool for researchers to compare structuresto this day. Shortly after, Desiraju introduced the term \u201csupramolecularsynthon\u201d to represent a structural unit or building block incrystal structures.19 Supramolecular synthonswere subclassified by us in 2003 into supramolecular homosynthons,between the same functional groups and supramolecular heterosynthons and between differentbut complementary functional groups .14 The understandingof supramolecular synthons, their geometries, and their frequencyof occurrence in the Cambridge Structural Database (CSD) is key tothe rational design of novel multicomponent crystal forms such ascocrystals and hydrates.20Certain intermolecular interactions,such as neutral and charge-assistedhydrogen bonds, and coordination bonds, are amenable to crystal engineeringstudies thanks to their relatively high strength, predictability,ubiquity, and directionality.21 Cocrystals are of interestin part because they can be readily accessible and amenable to crystalengineering. Cocrystals include molecular cocrystals22 (MCCs), which are comprised of two or more nonvolatileneutral molecules (coformers), and ionic cocrystals23 (ICCs), which involve at least one coformer that is a salt.ICCs are necessarily sustained by charge-assisted hydrogen bonds or,if metal ions are involved, coordination bonds.25 ICCs are therefore comprised of at least three components, a cation,an anion, and an additional molecule or salt. Given that MCCs of acompound typically offer a single variable, the coformer, ICCs canexhibit greater diversity in terms of composition and physicochemicalproperties compared to MCCs.4 Additionally,if an ICC is formed in which one coformer is an API and another isan API salt, then this type of ICC provides an opportunity to generatelow-dosage solid forms since the biologically active component(s)represent most of the mass of the ICC. Although ICCs are generallyless studied than MCCs, the first cocrystal involving sodium chlorideand urea was an ICC26 and, to our knowledge,at least five ICCs have been selected and developed for use in marketeddrug products: Depakote 27 in 1983; Lexapro 28 in 2002; Odomzo 29 in 2015; Entresto 30 in 2015; Seglentis (celecoxib and tramadol hydrochloride)31 in 2021.Cocrystalsare solids that are crystalline single-phase materialscomprised of two or more different molecular and/or ionic compoundsgenerally in a stoichiometric ratio that are neither simple saltsnor solvates.2+ and l-ascorbicacid, and inhibit the formation of oxygen free radicals.33 Statistically, ca. 8.7% of approved small-molecule drugs containedin DrugBank Online (Version 5.1.9.)34 arephenolic drugs, and phenol groups are present in ca. 10.1% of single-componentbiologically active compounds archived in the CSD (Scheme S1). However, many such compounds exhibit low aqueoussolubility and would therefore be classified as BCS class II.35 Their efficacy would therefore be hindered andan otherwise promising drug molecule could be rendered unsuitablefor use in an orally delivered drug product.37 Molecular cocrystals are now well known to modulate the physicochemicalproperties of phenolic compounds including solubility,38 bioavailability,39 stability,40 and mechanical properties41 while preserving their inherent biological activity.Phenolic drug molecules and nutraceuticalcompounds are of topicalinterest because they can exhibit biological effects such as antioxidantand anti-inflammatory activity, inhibit lipid peroxidation initiatedin rat brain homogenates by Fe20 carboxylate moieties,42 and aromatic nitrogen bases.43 Conversely,the phenol\u2013phenolate (PhOH\u00b7\u00b7\u00b7PhO\u2013) interaction is considered to be a strong hydrogen bond. In solution,PhOH\u00b7\u00b7\u00b7PhO\u2013 systems known as homoconjugatedcomplexes readily form as measured by their conjugated constants.44 PhOH\u00b7\u00b7\u00b7PhO\u2013 complexes can have a negative impact on titration experiments andplay an important role in enzyme catalysis.45 Buytendyk et al. reported the PhOH\u00b7\u00b7\u00b7PhO\u2013 interaction energy in the gas phase to be as high as 26\u201330kcal/mol, 60% of that of the HF\u00b7\u00b7\u00b7F\u2013 interaction, the strongest hydrogen bond known.46 Crystal structures involving PhOH\u00b7\u00b7\u00b7PhO\u2013 interactions archived in the CSD were mainly studiedfor structural insight47 or applicationssuch as organic synthesis (Kolbe\u2013Schmitt synthesis)49 and NLO.50 As such, there remains a lackof systematic crystal engineering studies on ICCs containing the PhOH\u00b7\u00b7\u00b7PhO\u2013 supramolecular heterosynthon.With respect to crystal engineering, phenols offer medium hydrogen-bonddonor strength and are established as being able to form supramolecularheterosynthons with hydrogen-bond acceptors such as chloride anions,\u2013 supramolecular heterosynthons. Specifically, phenoland four other substituted phenol derivatives were selected as modelcompounds and reacted with tetraalkylammonium hydroxides and potassiumhydroxide (\u2013 and phenol\u2013phenol (PhOH\u00b7\u00b7\u00b7PhOH)interactions in the obtained ICCs were examined using CrystalExplorer.In this contribution,we address ICC formation involving phenolsthrough a systematic CSD and experimental study that explores PhOH\u00b7\u00b7\u00b7PhOydroxide 1.In addPHNTMA; phenol\u00b7phenolate\u00b7potassium, PHNKOH; 1,3,5-benzentriol\u00b71,3,5-benzentriolate\u00b7tetramethylammonium, PGNTMA; 1,3,5-benzentriol\u00b71,3,5-benzentriolate\u00b7tetraethylammonium, PGNTEA; 4-isopropylphenol\u00b74-isopropylphenolate\u00b7potassium, IPPKOH; 4-isopropylphenol\u00b74-isopropylphenolate\u00b7tetrapropylammonium, IPPTPA; 4-isopropylphenol\u00b74-isopropylphenolate\u00b7tetrabutylammonium, IPPTBA; 4-methoxyphenol\u00b74-methoxyphenolate\u00b7tetrabutylammonium, MOPTBA; and resorcinol\u00b7resorcinolate\u00b7tetramethylammonium, RESTMA.A library of cocrystal formers containing five phenolderivatives,four organic bases, and one inorganic base was chosen for this study1. CocrysAll reagents and chemicals were purchasedfrom Sigma-Aldrich and used without further purification. Single crystalsof ICCs were obtained via slow evaporation of stoichiometric amountsof starting materials in a range of solvents at room temperature andharvested from solution before complete evaporation of their motherliquors had occurred.PHN andTMA were dissolved in 10 mL of acetonitrile (MeCN).The solvent was evaporated under vacuum until a viscous liquid wasobtained. Colorless crystals were harvested after exposing the viscousliquid to ambient conditions overnight.PHN and KOH were dissolved in 1 mL of methanol (MeOH). Thesolution was slowly evaporated at room temperature, and colorlesscrystals were obtained after 1 day.PGN and TMA were dissolved in 1 mL of MeOH. The solutionwas slowly evaporated at room temperature, and colorless crystalswere obtained after 1 day.PGN andTEA were dissolved in 1 mL of MeOH. The solutionwas slowly evaporated at room temperature, and colorless crystalswere obtained after several hours.2O) were dissolved in 1 mL of MeOH. Thesolution was slowly evaporated at room temperature, and colorlesscrystals were obtained after 1 day.IPP and KOH were dissolved in 1 mL of MeCN. The solutionwas slowly evaporated at room temperature, and colorless crystalswere obtained after 1 day.IPP and TPA were slurried in 1 mL of H2O overnight. The undissolved solid was isolated by filtration anddried in an oven. Colorless single crystals were obtained by dissolvingthe solid in MeOH, followed by slow evaporation at room temperature.IPP andTBA , followed by slow evaporation at room temperature.MOP and TBA were slurried in 2 mL of HRES , IPP ,and TMA were dissolved in 1 mLof MeOH. The solution was slowly evaporated at room temperature, andcolorless crystals were obtained after 1 week.All PXRD data werecollected on an Empyrean diffractometer with the followingexperimental parameters: Cu K\u03b1 radiation (\u03bb = 1.54056\u00c5); 40 kV and 40 mA; scan speed 8\u00b0/min; step size 0.05\u00b0,2\u03b8 = 5\u201340\u00b0.51 Structures were solved using SHELXT and refinedusing SHELXL contained in Olex2.52 Reflectiondata for the nonhydrogen atoms were refined anisotropically. All hydrogenatoms bonded to carbon were placed geometrically and refined using a ridingmodel with isotropic thermal parameters: Uiso(H) = 1.5Ueq(\u2212CH3), Uiso(H) = 1.2Ueq(\u2212CH), Uiso(H) = 1.2Ueq(\u2212CH2). Hydrogen atoms on water (AFIX 5) and methanol (AFIX 147)were calculated geometrically and refined using a riding model withisotropic thermal parameter: Uiso(H) =1.5Ueq(\u2212OH). The hydrogen atomsof phenolic hydroxyl groups were located from electron density differencemaps and included in the refinement process using a riding model with Uiso(H) = 1.2Ueq(\u2212OH).Single-crystal data are presented in Crystal structureswere determined by single-crystal X-ray diffraction(SCXRD) with either Cu K\u03b1 (\u03bb = 1.5418 \u00c5) radiationor Mo K\u03b1 (\u03bb = 0.71073 \u00c5) radiation and a Bruker D8Quest fixed-chi diffractometer equipped with a Photon 100 detectorand the nitrogen-flow Oxford Cryosystem attachment. Unit cell determination,data reduction, and absorption correction (multiscan method) wereconducted using the Bruker APEX3 suite with implemented SADABS software.R factor \u2264 0.05, no disorder andsingle-crystal structure only) was conducted to find crystal structuresthat contain PhOH\u00b7\u00b7\u00b7PhO\u2013 interactions.The following parameters were addressed by the search: (1) numberof structures that exhibit phenol\u2013phenolate interactions excludingsingle-component structures, salts, and zwitterions; (2) number ofstructures that form a PhOH\u00b7\u00b7\u00b7PhOH supramolecular homosynthon;(3) number of structures that contain a phenol group or a phenolateanion with the restriction that only C, H, and O were considered assubstituents on the phenyl ring . The distance distribution of phenol\u2013phenolateand phenol\u2013phenol hydrogen bonds and the bond length distributionof the C\u2013O bond of the phenol or phenolate group were plottedbased on data from Searches 1, 2, and 3. The schematic structuresused in searches 1, 2, and 3 are presented in ACSD survey level in the CrystalExplorerprogram package followed by geometry optimization carried out usingthe CASTEP module with GGA-type PBE functional contained in MaterialsStudio 8.0. The total interaction energy was partitioned into electronic(E_ele), polarization (E_pol), dispersion (E_dis), and repulsion (E_rel)components.\u2013 interactions are present. Of these entries, 69 (44.2%)are single-component compounds with phenol and phenolate moietiesin the same molecule; 30 entries (19.2%) are salts, and 15 (9.6%)are zwitterionic cocrystals or solvates composed of a zwitterion anda neutral molecule; 42 (26.9%) structures meet the definition of anICC that contains a phenol molecule, phenolate anion, and cation.However, of these 42 cocrystals, only one paper54 classifies them as cocrystals (refcode: JICKIS).Our CSD search retrieved 156 entries in which PhOH\u00b7\u00b7\u00b7PhO\u2013 O\u00b7\u00b7\u00b7O\u2013 bond distance determined from 42 ICC structures rangedfrom 2.4195(52)\u20132.6599(24) \u00c5 with an average of 2.528\u00b1 0.08\u00c5. A histogram of the O\u00b7\u00b7\u00b7O\u2013 distance distribution for PhOH\u00b7\u00b7\u00b7PhO\u2013 interactions is presented in The PhOH\u00b7\u00b7\u00b7PhOO\u2013H\u00b7\u00b7\u00b7Obond distances of PhOH\u00b7\u00b7\u00b7PhOH interactions determinedfrom search 2 ranged from 2.5218(28)\u20133.0381(45) \u00c5, averaging2.812 \u00b1 0.103 \u00c5. A histogram of the O\u2013H\u00b7\u00b7\u00b7Ohydrogen-bond distance distribution for PhOH\u00b7\u00b7\u00b7PhOHinteractions is presented in \u2013 bonds range from 1.2341(27) to 1.3776(95) \u00c5 witha mean of 1.307 (0.023) \u00c5. The distribution plots in CSD search3 addressed the C\u2013O bond length in phenols vs phenolates. TheCSD was found to contain 8277 and 192 entries for phenols and phenolates,respectively. \u2013\u00b7\u00b7\u00b7PhOH; PhOH\u00b7\u00b7\u00b7PhOH;PhO\u2013\u00b7\u00b7\u00b7H2O/MeOH; PhOH\u00b7\u00b7\u00b7H2O/MeOH. Tetramethylammonium (TMA), tetraethylammonium (TEA),tetrapropylammonium (TPA), and tetrabutylammonium (TBA) cations exhibitno strong hydrogen bonds. Rather, coulombic forces occur with phenolateanions. The neutral or ionic nature of phenolic groups in the nineICCs was addressed through proton location56 from difference Fourier map inspection and C\u2013O bond lengths. Table S1 lists the C\u2013O bond lengths inthe neutral (C\u2013OH) and deprotonated (C\u2013O\u2013) moieties.Hydrogen bonds werefound to be present in each of the nine novel ICCs reported herein:PhO\u2013 and PhO\u2013\u00b7\u00b7\u00b7H2O hydrogen bonds. PHNTMA crystallized in the space group P21/c. Theasymmetric unit of PHNTMA contains three phenol molecules, three phenolateanions, three TMA cations, and three water molecules in the ratioof 1:1:1:1 (R1410(28) (\u2013 hydrogen bonds withinthe discrete unit exhibit O\u00b7\u00b7\u00b7O\u2013 bonddistances of 2.438(2), 2.434(2), and 2.466(2) \u00c5. At the (100)plane, adjacent discrete units are arranged vertically and interactvia TMA cations. The discrete units align along the a-axis.PHNTMA, IPPTPA, IPPTBA, and MOPTBA formed discreteadducts. PHNTMA and IPPTBA crystallized as hydrates sustained by PhOH\u00b7\u00b7\u00b7PhO 1:1:1:1 2a. Four 1410(28) 2b. The rP21/c (R42(8) hydrogen-bondedring with each molecule interacting with an extra IPP molecule outsidethe ring to generate a discrete unit (\u2013 hydrogen bond with O\u00b7\u00b7\u00b7O\u2013 = 2.528(3) \u00c5. IPP serves as a hydrogen-bond donor and is hydrogen-bondedto a water molecule in each motif. The discrete units are connectedby TBA cations and aligned along the a-axis.IPPTBA crystallized as a 2:1:1:1 ICC of 4-isopropylphenol, 4-isopropylphenolate,TBA, and water in the space group P21/c 2c. Two 4ete unit 2d. The rP1\u0305 with one (Z\u2032 = 1) and two (Z\u2032 = 2) formula units in the asymmetric unit, respectively.In both structures, two discrete hydrogen-bonding assemblies existin each asymmetric unit, forming two different motifs C32(7) and C21(5) in IPPTPA (C32(7) motifs in MOPTBA \u00c5, 2.562(2) \u00c5, 2.571(3) \u00c5, 2.589(3) \u00c5; O\u00b7\u00b7\u00b7O\u2013 distances in MOPTBA: 2.5920(16) \u00c5, 2.4413(17)\u00c5, 2.4369(17) \u00c5, 2.6111(17) \u00c5). CH\u00b7\u00b7\u00b7Ointeractions and cations link the discrete units.IPPTPA and MOPTBA both crystallized in space group n IPPTPA 3a andtwn MOPTBA 3b. These48 Herein,single crystals of PHNKOH were grown from MeOH and are isostructuralwith HERCIQ but with a lower unit cell volume (4359.37 vs 4494.18\u00c53), presumably due to data collection at 150 K vsroom temperature. IPPKOH crystallized in P21/n. Althoughthey adopt different space groups, PHNKOH and IPPKOH both exhibitasymmetric units comprising three phenol molecules, one potassiumcation, and one phenolate anion in a 3:1:1 ratio. Each potassium cationadopts octahedral coordination geometry with five oxygen atoms fromphenol molecules (or phenol moieties of IPP), and the sixth coordinationsite is occupied by the \u03c0 system of a phenolate anion (S1a). The 1D coordination polymers in PHNKOHand IPPKOH form zigzag chains (S1b) that are supportedby three nonequivalent charge-assisted hydrogen bonds between eachphenolate moiety and three phenol moieties (O\u00b7\u00b7\u00b7O\u2013 in PHNKOH = 2.5276(18) \u00c5, 2.6052(19) \u00c5,2.6315(19) \u00c5; O\u00b7\u00b7\u00b7O\u2013 in IPPKOH= 2.541(2) \u00c5, 2.617(2) \u00c5, 2.546(3) \u00c5).PHNKOH and IPPKOH form one-dimensional (1D) coordinationpolymers.The structure of PHNKOH and its 2:1:1 variant (CSD Refcode HEQZUY) were previouslyreported as phenol solvates of potassium phenolate from high-resolutionPXRD data by Dinnebier et al.RESTMA isa 1:1:1 ICC of resorcinol, resorcinolate,and TMA hydrogen-bonded networks. Deprotonated resorcinol serves as ahydrogen-bond donor and acceptor since only one hydroxyl group isdeprotonated. The phenolate moieties of the resorcinolate anion formthree charge-assisted PhOH\u00b7\u00b7\u00b7PhO\u2013 hydrogen bonds. The resorcinolate anions interact with each otherto form a chain supported by PhOH\u00b7\u00b7\u00b7PhO\u2013 hydrogen bonds. These interactions result in perpendicular chainsbridged by resorcinol molecules, resulting in grid-like networks and one water molecule (PhO\u2013\u00b7\u00b7\u00b7H2O). Two phenolate moieties hydrogen-bond to phenolic groupsvia PhOH\u00b7\u00b7\u00b7PhO\u2013 hydrogen bonds ring motif to expandinto a ladder-like plane composed of rails hydrogen-bonded networks through PhO\u2013\u00b7\u00b7\u00b7MeOHand PhOH\u00b7\u00b7\u00b7MeOH hydrogen bonds, respectively also feature PhOH\u00b7\u00b7\u00b7PhOHsupramolecular homosynthons. The two most common motifs observed arethose in which a phenolate moiety interacts with two or three hydroxyl donors . Phenols are generally more acidic than aliphaticalcohols but less acidic than most carboxylic acids. This study presentsa strategy to form ICCs of phenols and their conjugate bases by PhOH\u00b7\u00b7\u00b7PhO\u2013 interactions to drive and sustain crystal packing.It is a well-recognized rule of thumb for salt and cocrystal screeningthat if \u0394pKa (\u0394pKa = pKa (base)-pKa (acid)) is >3, then a salt is formed and that if \u0394pKa is <0, then a cocrystal is favored.57 In this work, \u0394pKa is >3 for all bases used, and as such, proton transferwasanticipated even though proton transfer could have been affected byfactors such as solvent and the composition of starting materials.In this study, several solvent systems were used for ICC screening.Except for PHNTMA, only the use of MeOH or H2O as solventafforded high-quality single crystals suitable for SCXRD data collection.Poor quality crystals or gel-like phases were produced using ethanol,2-propanol, or acetone.As detailed above, charge-assisted PhOH\u00b7\u00b7\u00b7PhO\u2013 and PhOH\u00b7\u00b7\u00b7PhOHinteractions in the nine new ICCs reported herein are tabulated in Table S2 and presented as a scatter plot in Table S2), and the average strengthis presented in Selectedhydrogen bond lengthsfor PhOH\u00b7\u00b7\u00b7PhO\u2013 and PhOH\u00b7\u00b7\u00b7PhOH hydrogen-bond distancesin the ninenovel ICCs fall in the \u201cred zone\u201d and \u201cgreenzone\u201d, respectively. These distance zones are based upon O\u00b7\u00b7\u00b7O\u2013 distance ranges in PhOH\u00b7\u00b7\u00b7PhO\u2013 and PhOH\u00b7\u00b7\u00b7PhOH hydrogen bonds (\u00c5), respectively,determined from crystal structures archived in the CSD. PhOH\u00b7\u00b7\u00b7PhO\u2013 hydrogen bonds were found to exhibit relatively shortdistances (2.427(2)\u20132.658(2) \u00c5) and angles nearing 180\u00b0,whereas PhOH\u00b7\u00b7\u00b7PhOH hydrogen-bond distances were longer(2.825(3)\u20132.6289(19) \u00c5) and angles were more acute (149(5)\u2013177(8)\u00b0).Some charge-assisted PhOH\u00b7\u00b7\u00b7PhO\u2013 hydrogenbonds have relatively long O\u00b7\u00b7\u00b7O\u2013 distances or small angle values (2.6315(19) \u00c5, 161(3)\u00b0in PHNKOH; 2.617(2) \u00c5, 158(8)\u00b0 in IPPKOH; 2.726(3) \u00c5,180(7)\u00b0 in PGNTEA). These parameters are likely affected by thedifferent crystal packing environment in each crystal structure. Nevertheless,as expected, charge-assisted PhOH\u00b7\u00b7\u00b7PhO\u2013 hydrogen bonds were found to be consistently shorter than PhOH\u00b7\u00b7\u00b7PhOHhydrogen bonds. Further, as presented in \u2013 interactions were calculated to offer ca. 3 times the energy ofPhOH\u00b7\u00b7\u00b7PhOH interactions, with average energies of91.6 \u00b1 8.1 and 28.53 \u00b1 0.67 kJ/mol, respectively. As alsodetailed in \u2013 and PhOH\u00b7\u00b7\u00b7PhOHhydrogen-bond strength. Specifically, for PhOH\u00b7\u00b7\u00b7PhOHinteractions, the electrostatic contribution comes from dipole\u00b7\u00b7\u00b7dipoleinteractions, whereas for PhOH\u00b7\u00b7\u00b7PhO\u2013 interactions, the phenolate oxygen and phenolic hydrogen are keyto the electrostatic contribution. The greater electrostatic energiesin PhOH\u00b7\u00b7\u00b7PhO\u2013 can therefore be attributedto coulombic forces. Polarization contributes to total energy forPhOH\u00b7\u00b7\u00b7PhO\u2013, while it is negligiblefor PhOH\u00b7\u00b7\u00b7PhOH. A small dispersion contribution wascalculated for all hydrogen bonds. Repulsion forces for PhOH\u00b7\u00b7\u00b7PhO\u2013 were consistently calculated to be greater than forPhOH\u00b7\u00b7\u00b7PhOH hydrogen bonds. Overall, although repulsiveforces for the charge-assisted hydrogen bonds are relatively high,the increase is more than offset by increases in permanent chargeand electronic and polarization forces. These high intermolecularinteraction energies and desirable hydrogen-bond geometry featuresvalidate that charge-assisted PhOH\u00b7\u00b7\u00b7PhO\u2013 hydrogen bonds are a strong and reliable interaction to enable theformation of ionic cocrystals.As shown by the scatter plot of \u2013 supramolecular heterosynthon is understudied in thecontext of crystal engineering of ICCs. To address this issue, ninenovel ICCs involving phenol and phenol-substituted compounds wereformed by reaction with bases and their crystal structures were determinedby SCXRD. Analysis of the resulting crystal structures revealed thatall nine ICCs are sustained by the phenol\u2013phenolate supramolecularheterosynthon with O\u00b7\u00b7\u00b7O\u2013 distancesranging from 2.427(2) to 2.658(2) \u00c5. A computational study validatedthe robustness of the charge-assisted PhOH\u00b7\u00b7\u00b7PhO\u2013 supramolecular heterosynthon vs. the phenol\u2013phenolsupramolecular homosynthon from an energetic perspective. Based onthese results, we consider that the PhOH\u00b7\u00b7\u00b7PhO\u2013 supramolecular heterosynthon is suitable for the reliableformation of ICCs of phenolic compounds, which are prevalent in drugmolecules and nutraceuticals. That phenols form ICCs with their conjugatebases is important from a pharmaceutical perspective as it means thatthe mass of a dosage form can be close to that of the parent phenol.Future studies will focus on pharmaceutical and nutraceutical cocrystals,their physicochemical properties, and polymorphism.Phenolgroups are found in 8.7% of approved small-molecule drugsin the DrugBank database and in 10.1% of bioactive single-componentcompounds deposited in the CSD. Our CSD survey revealed that the PhOH\u00b7\u00b7\u00b7PhO"} +{"text": "The packing of the title compound is consolidated by C\u2014H\u22efO inter\u00adactions. 23H19NO5, the cyano group adopts an axial orientation and the ester group an equatorial orientation. The dihedral angle between the pendant phenyl group and the benzene ring of the fused-ring system is 25.97\u2005(8)\u00b0. Intra\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds are observed and the packing is consolidated by C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, C H-benzo[c]chromen-6-ones or 3,4,5,6-dibenzo-\u03b1-pyran\u00adones) form an important group of biologically active natural products that occur in bacteria, fungi, lichens, higher plants and animal waste \u00b0. The Cremer\u2013Pople puckering parameters of the O1/C9/C10/C13\u2013C15 and C14/C15/C17\u2013C20 rings indicate half-chair conformations in each case with puckering amplitudes Q = 0.359\u2005\u00c5; \u03b8 = 104.52\u00b0; \u03c6 = 9.27\u00b0 and Q = 0.49\u2005\u00c5; \u03b8 = 134.17\u00b0; \u03c6 = 327.35\u00b0, respectively. The O atom attached to C17 is stabilized in its enol (hy\u00addroxy) form, presumably as a result of forming a strong intra\u00admolecular hydrogen bond to O2. The packing is consolidated by weak C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions , phenyl\u00adboronic acid , K3PO4 and Pd(PPh3)4 in degassed 1,4-dioxane (10\u2005mL) was stirred at 100\u00b0\u2005C for 12\u2005h under nitro\u00adgen. After completion of the coupling reaction (TLC), the mixture was cooled to room temperature, diluted with di\u00adchloro\u00admethane and deca\u00adnted. The residue was extracted with DCM (10\u2005mL \u00d7 2) twice. The solvent was removed from the combined DCM layers and the residue was subjected to column chromatography on silica gel (100\u2013200 mesh) by using increasing amounts of ethyl acetate in hexane (5% to 15%) as eluent to afford the title compound as a light-yellow solid in 90% yield (84\u2005mg); Rf = 0.4 ; m.p. 155\u2013158\u00b0\u2005C. A sample suitable for single-crystal X-ray analysis was obtained by recrystallization the 50\u2005mg of the solid from a mixture of 1\u2005mL of distilled chloro\u00adform and 0.5\u2005mL of distilled methanol.A mixture of ethyl 10-cyano-7-hy\u00addroxy-6-oxo-3-{[(tri\u00adfluoro\u00admeth\u00adyl)sulfon\u00adyl]\u00adoxy}-8,9,10,10Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314622001997/hb4401sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622001997/hb4401Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622001997/hb4401Isup3.cmlSupporting information file. DOI: 2153368CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-isopropylacrylamide) (pNIPAM) and poly(N-tert-butylacrylamide) (pNTBAM) hydrogels were assessed in terms of their chemical differences, mechanical strength, internal architecture, porosity and capacity to support cell viability, migration, and differentiation. pNTBAM polymerized with a Young\u2019s modulus of up to 371\u2009\u00b1\u200931\u00a0kPa compared to the more flexible pNIPAM, 16.5\u2009\u00b1\u20090.6\u00a0kPa. Viability testing revealed biocompatibility of both hydrogels with significantly increased cell numbers observed in pNTBAM (500\u2009\u00b1\u200995 viable cells/mm2) than in pNIPAM (60\u2009\u00b1\u20093 viable cells/mm2) (P\u2009\u2264\u20090.05). Mineralization determined through alkaline phosphatase (ALP) activity, calcium ion and annexin A2 markers of mineralization) and osteogenic behaviour (collagen I expression) were supported in both hydrogels, but to a greater extent in pNTBAM. pNTBAM supported significantly elevated levels of chondrogenic markers as evidenced by collagen II and glycosaminoglycan expression in comparison to little or no evidence in pNIPAM (P\u2009\u2264\u20090.05). In conclusion, structurally similar, chemically distinct, acrylamide hydrogels display variable capacities in supporting osteochondral cell behaviours. These systems demonstrate spatial control of cell interaction through simple changes in monomer chemistry. Fine control over chemical presentation during the fabrication of biomaterial implants could lead to greater efficacy and targeted regeneration of semi-complex tissues.Regeneration solutions for the osteochondral interface depth are limited, where multi-material implants have the potential to delaminate affecting the regeneration process and impacting the final integrity of tissue interface. Here we explore regionally mixed hydrogel networks, presenting distinct chemical features to determine their compatibility in supporting osteogenic or chondrogenic cell behaviour and differentiation. Poly( Furthermore, material characteristics influence the cellular capacity to perform biological functions and N-tert-butylacrylamide (NTBAM) have been described as co-polymers with potential to stimulate tissue regeneration including that of bone and cartilage was used to link the polymer network. Ammonium persulfate (APS) 10% w/v initiator was used alongside N,N,N\u2032,N\u2032-tetramethylethylenediamine (TMED) 2.6% v/v as an accelerator. NIPAM formed a transparent soft gelatinous structure requiring 2\u20133\u00a0min to form after rapid addition of the initiator. NTBAM was formed in a 1:1 combination of water and methanol heated to 37\u00a0\u00b0C. pNTBAM hydrogel formation required 10\u201315\u00a0min to completely gelate.Hydrogels were prepared by liquid phase atom transfer radical polymerization (Matyjaszewski Hydrogels were observed using a bench-top Hitachi S4500 scanning electron microscope (SEM) at 5\u00a0kV. Hydrogels were frozen at \u2013\u00a020\u00a0\u00b0C, freeze dried, mounted onto carbon tape, and gold sputter coated. Pore size measurements were performed using Image J, with over 100 features measured per sample and 3 repeat samples measured.Measurement of water contact angle was performed using a \u201cTheta Lite Attension One Attension version 2.4\u201d system. Samples were placed on a glass petri dish and compressed with a cover slip to acquire a flat surface while being dried at 70\u00a0\u00b0C in an oven. Water droplets, measured at 1 \u03bcL, were slowly placed onto test surfaces. Four measures were collected from each sample out of 4 samples.Compressive strengths were measured with the BOSE Electroforce system equipped with a 20\u00a0N loading cell and cross head speed of 0.05\u00a0mm/s. The samples were cylindrical in shape with dimensions of 4.5\u20135.0\u00a0mm height and 9.4\u201311.5\u00a0mm diameter. The load was applied until strain reached 90%. Compressive strength was determined from the maximum load of the applied stress\u2013strain curve. Four samples of each hydrogel were tested, and an average obtained.L-glutamine. Culture environments were 37\u00a0\u00b0C in 5%\u00a0CO2 culture incubators with media changed every 5\u00a0days. Primary human osteoblasts (hOB) and primary human chondrocytes (hCH) were obtained commercially from Promo Cell\u00ae and cultured under the conditions described above. Routine trypsinization protocols were followed for cell passaging and immortalized (human telomerase transcriptase (hTERT) transduced) chondrocytes (OK3H) or chondrogenic induction supplements .Differentiation was induced by addition of either osteogenic induction supplements . The major components of the assay kit are calcein AM and ethidium homodimer-1 reagents. Calcein AM identifies the presence of live cells by detecting intracellular esterase activity and cell membrane integrity. Live/dead staining solution was prepared by mixing both reagents with PBS at the following rates; 1:200 of calcein AM and 1:50 of ethidium homodimer-1 in PBS. The cells viability profile was assessed after 21\u00a0days of cell culturing. The hydrogel samples were removed from media and washed 3 times with PBS. Next, these samples were incubated with 1\u00a0mL/sample of live/dead staining solution for 30\u00a0min at room temperature protected from light. Hydrogels were viewed under an Olympus U-TBI90 confocal microscope to observe the viable (green fluorescence) and the dead (red fluorescence) cells. The viability of cells upon hydrogel samples was identified by calculating the number of live and dead cells per specific regions of each sample. The number of live cells (green) and dead cells (red) were counted over a 1 mm2 area for a maximum of 5 regions of a captured\u2009\u00d7\u20094 microscopic images obtained for individual samples and the average was taken. The whole process was carried out using the cell counting tool of Image J software.Viability of cells was examined with the live/deadCell migration was established after 21\u00a0days culture within the hydrogels. Hydrogel samples were removed from media, washed 3 times with PBS, and fixed in 10% paraformaldehyde for 30\u00a0min at room temperature. Then, each sample was incubated with 1\u00a0mL DAPI stain solution for 30\u00a0min at room temperature. Samples were then scanned across the z-axis using a 2\u00a0\u00b5m step size reaching a maximum of 150 slices of sample down from the top layer. The scanned distance for each sample was set to a maximum of 300\u00a0\u00b5m starting from the surface. Scanned files were processed via Image J software. The depth location was determined by nuclei localisation within the depth of the hydrogel.Primary antibodies for collagens type I, II, and X confirmed differentiation. Visualization was enabled through appropriate secondary antibodies; Goat Anti-Rabbit IgG H&L, Abcam cat. No. 6718 and 6717, respectively). Hydrogel samples were fixed with 10% formaldehyde for 30\u00a0min at room temperature. Samples were blocked with 5% bovine serum albumin (BSA) in PBS for 2\u20133\u00a0h at 4\u00a0\u00b0C. This was followed by sample incubation with primary antibody solution overnight at 4\u00a0\u00b0C. The primary solution was prepared by mixing primary antibody with 5% BSA in PBS at 1:200 ratio. The primary solution was then aspirated, and samples washed 4 times with 1% BSA in PBS solution. Samples were then incubated with secondary antibody (conjugated with either FITC or TRITC) in 5% BSA in PBS solution at 1:200 ratio at 4\u00a0\u00b0C for 4\u00a0h in the dark. Samples were then washed with 1% BSA in PBS followed by additional PBS washes. Nuclear staining was through DAPI stain for 30\u00a0min at room temperature followed by PBS washes. Hydrogels were imaged with an Olympus U-TBI90 laser fluorescent confocal microscope.Extraction of calcium minerals from hydrogel samples was performed using 0.5\u00a0M HCl solution. Hydrogels were first fixed with 10% paraformaldehyde in phosphate buffer saline (PBS) for 30\u00a0min at room temperature. Each hydrogel sample was incubated with 500 \u00b5L of HCl extraction solution at room temperature overnight on a rotary shaker. Calcium mineral was assessed using the calcium colorimetric assay kit (Sigma Aldrich MAK022) following manufacturer\u2019s instructions with calcium ions present in samples Morin detectedGAGs matrix proteins were assessed by dimethylmethylene blue (DMMB) assay . Hydrogels were digested in papain lysis buffer at 60\u00a0\u00b0C overnight. Cell lysates were then assessed for collagens I, II and annexin A2 subsequent to the application of the assay protocol specific for each marker. Absorbance was measured with a Synergy II BioTek plate reader.P\u2009\u2264\u20090.05) Fig.\u00a0B. pNTBAMP\u2009\u2264\u20090.05). Pore size increased with lowering hydrogel monomeric concentrations for both hydrogels (p\u2009\u2264\u20090.05). For pNIPAM, 0.058\u00a0g/mL and 0.04\u00a0g/mL concentration resulted in mean pore diameters of 29.9\u2009\u00b1\u20097.7\u00a0\u00b5m and 35.7\u2009\u00b1\u20099\u00a0\u00b5m, respectively. Similarly, with pNTBAM, 0.058\u00a0g/mL and 0.04\u00a0g/mL concentrations resulted in mean pore diameters of 16.3\u2009\u00b1\u20093.6 \u00b5mand 21.2\u2009\u00b1\u20095\u00a0\u00b5m, respectively .Osteosarcoma MG63 and chondrogenic OK3H cells seeded onto pNIPAM samples formed distinct aggregates or clusters while conversely, with pNTBAM, they attach and spread out across the hydrogel surface Fig.\u00a0A, B. ThiP\u2009\u2264\u20090.05) and 0.058\u00a0g/mL (44.8\u2009\u00b1\u200919 pNIPAM \u00b5m) and 0.042\u00a0g/mL (pNIPAM) for primary cell experimentation. Further, based on identified properties (stiffer mass and high cellular density) pNTBAM was selected as a putative best choice for support of mineralization while the smaller pores, hydrophobicity, lower cellular density, cluster growth, and flexibility suggested pNIPAM as a putative choice for chondrogenic tissue growth.2) when compared to hOBs (243.6\u2009\u00b1\u200938.26 cells/ mm2) on pNTBAM (P\u2009\u2264\u20090.05). We again noted that pNIPAM displayed reduced cell numbers (58\u2009\u00b1\u20097.3 cells/mm2 for hOBs and 65\u2009\u00b1\u20093.3 cells/mm2) when compared to pNTBAM, . pNTBAM produced 2.09\u2009\u00b1\u20090.04 versus 0.37\u2009\u00b1\u20090.04\u00a0\u00b5g/\u00b5L calcium ions whilst pNIPAM produced the lower amount of 1.15\u2009\u00b1\u20090.04 versus 0.47\u2009\u00b1\u20090.06\u00a0\u00b5g/\u00b5L calcium ions (P\u2009\u2264\u20090.05). Collagen I levels produced by hOBs were significantly elevated in osteogenic media when compared to control samples (P\u2009\u2264\u20090.05) . Complementary to increases in collagen I and calcium ion production we noted that ALP activity levels were increased from 0.2 \u00d7 103\u2009\u00b1\u20090.1 \u00d7 103 for controls to 3.3 \u00d7 103\u2009\u00b1\u20090.6 \u00d7 103 U/\u00b5L for pNTBAM osteogenic samples (P\u2009\u2264\u20090.05) which were themselves in turn significantly elevated (P\u2009\u2264\u20090.05) when compared to pNIPAM values of 1.4 \u00d7 103\u2009\u00b1\u20090.2 \u00d7 103 U/\u00b5L versus 0.3 \u00d7 103\u2009\u00b1\u20090.1 \u00d7 103 U/\u00b5L in untreated controls (P\u2009\u2264\u20090.05) Fig.\u00a0A.Fig. 7OP\u2009\u2264\u20090.05). pNTBAM produced 31.3 \u00d7 103\u2009\u00b1\u20094.7 \u00d7 103 in osteogenic compared to 2.7 \u00d7 103\u2009\u00b1\u20091.8\u00a0ng/g control, whereas pNIPAM produced 17.2 \u00d7 103\u2009\u00b1\u20092.4 \u00d7 103 versus 2.3 \u00d7 103\u2009\u00b1\u20091.2 \u00d7 103\u00a0ng/g of controls when compared to pNTBAM (0.52\u2009\u00b1\u20090.06\u00a0\u00b5g/\u00b5L pNIPAM versus 0.14\u2009\u00b1\u20090.06\u00a0\u00b5g/\u00b5L for pNTBAM). Both were significantly lower (P\u2009\u2264\u20090.05) than hOB samples in osteogenic media. Hydrogels seeded with hCHs in chondrogenic media displayed increased collagen I levels when compared to controls (pNTBAM (85\u2009\u00b1\u200923 versus 26\u2009\u00b1\u20096\u00a0ng/g) and pNIPAM (133\u2009\u00b1\u200933 versus 78\u2009\u00b1\u200919\u00a0ng/g) (P\u2009>\u20090.05) Fig.\u00a0A. Howeve Fig.\u00a0A. P\u2009\u2264\u20090.05). pNIPAMhCHs samples were not significantly different compared to their controls and were significantly lower (P\u2009\u2264\u20090.05) than pNTBAM (1.15\u2009\u00b1\u20090.49\u00a0\u00b5g pNIPAM versus 25\u2009\u00b1\u20090.74\u00a0\u00b5g for pNTBAM chondrogenic samples).Chondrogenic hCHs samples revealed significant elevation in GAG expression for pNTBAM hydrogels (25\u2009\u00b1\u20090.7\u00a0\u00b5g versus 12.26\u2009\u00b1\u20091.5\u00a0\u00b5g control) (P\u2009\u2264\u20090.05). pNIPAM displayed no significant changes (224\u2009\u00b1\u2009112 versus 74.8\u2009\u00b1\u200964.7\u00a0ng/g control). pNTBAM, again, showed significant elevation when compared to pNIPAM chondrogenic samples (3105\u2009\u00b1\u2009282 versus 224\u2009\u00b1\u2009112\u00a0ng/g for pNIPAM) (pNIPAM) and poly(N-tert-butylacrylamide) (pNTBAM) hydrogels were produced by polymerization induced phase separation resulting from differing wettablity between monomer and polymer network, a widely applied method in the production of porous polymer scaffolds with chondrogenic differentiation. We conclude that when taken together, these unique polymer characteristics create a potentially tuneable platform for 3D osteochondral scaffold generation for application in future tissue engineered complex repair situations."} +{"text": "The structure of a copper(II) complex is described. 6H4O2N)2]\u00b72C9H6O6\u00b72H2O, the Cu2+ ion lies on a center of inversion and coordinates with symmetry related pyridine nitro\u00adgen and carboxyl oxygen atoms from two pyridine-2-carb\u00adoxy\u00adlic acid anions, giving rise to a square-planar coordination geometry. There are weak axial bonds between Cu and an O atom of a symmetry-related trimesic acid moieties [Cu\u22efO = 2.837\u2005(2)\u2005\u00c5] The Cu\u22efO weak inter\u00adactions and hydrogen bonds stabilize the whole structure.In the title complex, [Cu(C In the axial position, a very weak inter\u00adaction Cu1\u22efO3 [2.837\u2005(2)\u2005\u00c5] is observed. Inter\u00adestingly, the 1,4-bis\u00ad(3-pyrid\u00adyl)-2,3-di\u00adaza-1,3-butadiene ligand decomposed during the hydro\u00adthermal process and is oxidized into pyridine-2-carb\u00adoxy\u00adlic acid. According to our earlier research, the occurrence of oxidation may be caused by excess of CuII salt, which may act as an oxidative agent to promote the formation of the carboxyl group ligand and one crystal water mol\u00adecule Fig.\u00a01. The Cu23\u2212 ligands are short, there are no \u03c0\u2013\u03c0 inter\u00adactions because the inter-centroid distance between the two benzene rings is 5.4029\u2005(15)\u2005\u00c5, which is much larger than the normal \u03c0\u2013\u03c0 stacking distance of 3.3\u20133.8\u2005\u00c5. The shortest distance between the two carbon atoms (C1 and C1\u2032) is 3.379\u2005(4)\u2005\u00c5. The other C\u22efC distances of the two rings are longer than 3.94\u2005\u00c5. In addition, the distance between the centroid of one benzene ring and the C atoms of another is longer than 4.28\u2005\u00c5.In the crystal, C\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds Table\u00a01 and toge2\u00b72H2O in 5\u2005mL of distilled H2O was stirred for 10\u2005min in air, and then the mixture was turned into a Parr Teflon-lined stainless steel vessel and heated at 160\u00b0C for 60\u2005h. Dark-red crystals suitable for X-ray diffraction were obtained in a yield of 78% (based on CuCl2\u00b72H2O).A mixture of trimesic acid , 1,4-bis\u00ad(3-pyrid\u00adyl)-2,3-di\u00adaza-1,3-butadiene and CuClCrystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621006726/bv4039sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314621006726/bv4039Isup3.hklStructure factors: contains datablock(s) I. DOI: 2092720CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In thermophiles, tRNAs are heavily modified to maintain their thermal stability under extreme growth temperatures6. Here we identified 2\u2032-phosphouridine (Up) at position 47 of tRNAs from thermophilic archaea. Up47 confers thermal stability and nuclease resistance to tRNAs. Atomic structures of native archaeal tRNA showed a unique metastable core structure stabilized by Up47. The 2\u2032-phosphate of Up47 protrudes from the tRNA core and prevents backbone rotation during thermal denaturation. In addition, we identified the arkI gene, which encodes an archaeal RNA kinase responsible for Up47 formation. Structural studies showed that ArkI has a non-canonical kinase motif surrounded by a positively charged patch for tRNA binding. A knockout strain of arkI grew slowly at high temperatures and exhibited a synthetic growth defect when a second tRNA-modifying enzyme was depleted. We also identified an archaeal homologue of KptA as an eraser that efficiently dephosphorylates Up47 in vitro and in vivo. Taken together, our findings show that Up47 is a reversible RNA modification mediated by ArkI and KptA that fine-tunes the structural rigidity of tRNAs under extreme environmental conditions.Post-transcriptional modifications have critical roles in tRNA stability and function Reversible internal RNA phosphrylation contributes to thermal stability and nuclease resistance of tRNA, and cellular thermotolerance of hyperthermophiles.\u00a0 Thus far, about 150 types of RNA modification have been reported in various RNA molecules from all domains of life7. In particular, tRNAs contain the widest variety and largest number of modified nucleosides, with 80% of RNA modifications identified in tRNA molecules. Diverse RNA modifications are clustered in the anticodon loop, especially at positions 34 and 37 (refs. 8). These modifications have critical roles in stabilizing and modulating codon\u2013anticodon interactions on the ribosome, ensuring accurate and efficient protein synthesis. Many RNA modifications are also found in the tRNA body composed of the D-loop, T\u03a8C loop (T-loop) and variable loop (V-loop)10 confer conformational rigidity to the tRNA core region by fixing C3\u2032-endo ribose puckering11.Recent advances in epitranscriptomics research have demonstrated the chemical diversity and biological importance of RNA modifications)10 Fig. . These R6. 5-Methyl-2-thiouridine (m5s2U or s2T) is found at position 54 in the T-loop of tRNAs from thermophiles12. m5s2U54 adopts a rigid conformation with C3\u2032-endo ribose puckering, thereby stabilizing the tRNA body in high-temperature environments11. The 2-thiolation level of m5s2U54 increases as the growth temperature rises14. m5s2U54 contributes to the thermotolerance of Thermus thermophilus15. In Pyrococcus furiosus, the relative abundance of N4-acetylcytidine (ac4C) and its 2\u2032-O-methyl derivative (ac4Cm) were markedly increased with rising growth temperature14. ac4C is a prevalent modification that is present in tRNAs, rRNAs and other RNAs in hyperthermophilic archaea16. ac4C favours the C3\u2032-endo form and stabilizes tRNAs18. Loss of ac4C results in a growth defect in Thermococcus kodakarensis at high temperature, contributing to cellular\u00a0thermotolerance19. In Bacillus stearothermophilus, 2\u2032-O-methylation in tRNAs increases when the growth temperature rises20. Archaeosine (G+) is a unique 7-deazaguanosine derivative found at position 15 in the D-loop of archaeal tRNAs21. On the basis of quantum mechanics calculations, G+15 stabilizes the Levitt base pair with C48 (ref. 22). In line with this, biochemical and genetic studies have shown that G+ confers thermal stability to tRNAs and contributes to thermotolerance23.In thermophilic bacteria and archaea, unique RNA modifications contribute to the thermal adaptation of tRNAsp) in tRNAs, which, to our knowledge, is the first known instance of internal RNA phosphorylation. Biochemical, structural and genetic studies showed that Up47 is a reversible RNA modification and confers thermal stability to tRNA, thereby contributing to cellular thermotolerance.Here we report the identification of 2\u2032-phosphouridine 27 and mapped 13 types of RNA modification at 18 positions in this tRNA molecule at position 47 of the RNA fragments digested with RNases modification in class I tRNAs bearing U47 in the V-loop, but not in class II tRNAs with a long V-loop , with a low error value of 4.4\u2009millimass unit\u00a0, indicating that N324 is a phosphorylated uridine residue. This prediction explains why the N324 nucleoside was not detected in our nucleoside analysis and biochemical approaches Fig. .p47 is a thermophile-specific modification found in the tRNA core region, we investigated whether Up47 stabilizes the tertiary structure of tRNA. To this end, we treated S.\u2009tokodaii tRNAVal3 with yeast Tpt1p (2\u2032-phosphotransferase) to remove the 2\u2032-phosphate of Up47. We measured the melting temperature (Tm) of S.\u2009tokodaii tRNAVal3 with and without Up47 . According to our analysis of Up47 distribution in archaeal species nucleotide analysis. A pUpm5C dimer was clearly observed in the parental strain (wild type) of T.\u2009kodakarensis (KU216), but was absent in the \u0394tk2051 strain .To identify a gene responsible for Uies Fig. . Among tp47 Fig. . Among tp47 Fig. . We chosp47 Fig. . STK_095ain Fig. . Therefop47 in T.\u2009kodakarensis. The \u0394arkI strain grew as well as the wild-type strain (KU216) at the nearly optimal temperature of 83\u2009\u00b0C formation, because G+15 thermally stabilizes tRNAs and contributes to cellular thermotolerance19. We confirmed the absence of Up47 and G+15 in tRNAs from the double-knockout strain and examined in vitro Up47 formation. Up47 was efficiently reconstituted only in the presence of ATP Fig. , TkArkI The electrostatic surface potential showed a large positive area on one side of the TkArkI structure Fig. . The posTo characterize the conserved residues in TkArkI, we constructed 14 TkArkI mutants in which targeted residues were replaced by alanine Fig. . All mut33. Tpt1/KptA homologues are distributed across all domains of life35 in the presence of NAD+, with the results indicating that the 2\u2032-phosphate of Up47 was efficiently removed , because N.\u2009viennensis is a mesophilic archaeon and its ArkI homologue was predicted to have efficient activity in E.\u2009coli. The class I tRNA fraction prepared from this strain was subjected to shotgun analysis to detect the Up47 modification. We clearly detected four Up47-containing fragments derived from various E.\u2009coli tRNA species -inducible promotor and quantified the peak intensity of each Up47-containing fragment when TkKptA expression was induced by addition of 10 or 100\u2009\u03bcM IPTG and two class II tRNAs (tRNALeu4 and tRNASer3) Fig. but was 2+. Especially at high temperatures, RNA is rapidly degraded. Similarly to 2\u2032-O-methylation, the 2\u2032-phosphorylation of Up47 also serves to prevent tRNA degradation. This property partly explains the RNase resistance of tRNA conferred by Up47 with different conformations in the core region to fill in for the missing G46, forming the non-canonical base triple \u03a813\u2013G22\u2013C9 has a critical role in RNA metabolism and function as a reversible RNA modification45. If Up47 is a reversible modification, it is expected that tRNA function and stability are dynamically regulated by a writer and eraser, raising the possibility of epitranscriptomic regulation of tRNAs in translation. The mechanism closely resembles post-translational modification of proteins. Phosphorylation and dephosphorylation rapidly and dynamically control protein function48. Because tRNA is a stable molecule with a low turnover rate and long lifetime in the cell, it would be reasonable for tRNA function to be regulated by Up47 modification. We found efficient dephosphorylation of Up47 by TkKptA in vitro , T. Yokogawa (Gifu University) and H. Hori (Ehime University), respectively. Sulfolobus acidocaldarius (JCM no. 8929), Saccharolobus solfataricus (JCM no. 8930), Aeropyrum pernix (JCM no. 9820), Pyrobaculum oguniense (JCM no. 10595) and N.\u2009viennensis (JCM no. 19564) were obtained from\u00a0Japan Collection of Microorganisms, RIKEN BRC which is participating in the National BioResource Project of the MEXT, Japan.S.\u2009tokodaii and S.\u2009acidocaldarius were cultured at 80\u2009\u00b0C in JCM medium no. 165 consisting of 1\u2009g\u2009l\u20131 yeast extract, 1\u2009g\u2009l\u20131 casamino acids, 1.3\u2009g\u2009l\u20131 (NH4)2SO4, 0.28\u2009g\u2009l\u20131 KH2PO4, 0.25\u2009g\u2009l\u20131 MgSO4\u00b77H2O, 0.07\u2009g\u2009l\u20131 CaCl2\u00b72H2O, 2.0\u2009mg\u2009l\u20131 FeCl3\u00b76H2O, 1.8\u2009mg\u2009l\u20131 MnCl2\u00b74H2O, 4.5\u2009mg\u2009l\u20131 Na2B4O7\u00b710H2O, 0.22\u2009mg\u2009l\u20131 ZnSO4\u00b77H2O, 0.05\u2009mg\u2009l\u20131 CuCl2\u00b72H2O, 0.03\u2009mg\u2009l\u20131 Na2MoO4\u00b72H2O, 0.03\u2009mg\u2009l\u20131 VOSO4\u00b7H2O and 0.01\u2009mg\u2009l\u20131 CoSO4\u00b77H2O (adjusted to pH\u20092.5 with H2SO4). S.\u2009solfataricus was cultured at 80\u2009\u00b0C in JCM medium no. 171 consisting of 1\u2009g\u2009l\u20131 yeast extract, 2.5\u2009g\u2009l\u20131 (NH4)2SO4, 3.1\u2009g\u2009l\u20131 KH2PO4, 0.2\u2009g\u2009l\u20131 MgSO4\u00b77H2O, 0.25\u2009g\u2009l\u20131 CaCl2\u00b72H2O, 1.8\u2009mg\u2009l\u20131 MnCl2\u00b74H2O, 4.5\u2009mg\u2009l\u20131 Na2B4O7\u00b710H2O, 0.22\u2009mg\u2009l\u20131 ZnSO4\u00b77H2O, 0.05\u2009mg\u2009l\u20131 CuCl2\u00b72H2O, 0.03\u2009mg\u2009l\u20131 Na2MoO4\u00b72H2O, 0.03\u2009mg\u2009l\u20131 VOSO4\u00b7H2O and 0.01\u2009mg\u2009l\u20131 CoSO4\u00b77H2O (adjusted to pH\u20094.0 with H2SO4). A.\u2009pernix was cultured at 90\u2009\u00b0C in JCM medium no. 224 consisting of 1\u2009g\u2009l\u20131 yeast extract, 1\u2009g\u2009l\u20131 peptone, 1\u2009g\u2009l\u20131 Na2S2O3\u00b75H2O, 24.0\u2009g\u2009l\u20131 NaCl, 7.0\u2009g\u2009l\u20131 MgSO4\u00b77H2O, 5.3\u2009g\u2009l\u20131 MgCl2\u00b76H2O, 0.7\u2009g\u2009l\u20131 KCl and 0.1\u2009g\u2009l\u20131 CaCl2\u00b72H2O (adjusted to pH\u20097.0 with NaOH). P.\u2009oguniense was cultured at 90\u2009\u00b0C in JCM medium no. 165 with addition of 1.0\u2009g\u2009l\u20131 Na2S2O3\u00b75H2O (adjusted to pH\u20097.25 with NaOH). N.\u2009viennensis was cultured at 42\u2009\u00b0C in JCM medium no. 1004 consisting of 1\u2009g\u2009l\u20131 NaCl, 0.5\u2009g\u2009l\u20131 KCl, 0.4\u2009g\u2009l\u20131 MgCl2\u00b76H2O, 0.2\u2009g\u2009l\u20131 KH2PO4, 0.1\u2009g\u2009l\u20131 CaCl2\u00b72H2O, 1.0\u2009ml\u2009l\u20131 modified trace element mixture , 1.0\u2009ml\u2009l\u20131 vitamin solution ), 1.0\u2009ml\u2009l\u20131 7.5\u2009mM EDTA\u00b7Na\u00b7Fe(III) solution (pH\u20097.0), 2.0\u2009ml\u2009l\u20131 1\u2009M NaHCO3 solution, 10\u2009ml\u2009l\u20131 HEPES solution (238.4\u2009g\u2009l\u20131 HEPES (free acid) and 24\u2009g\u2009l\u20131 NaOH), 1.0\u2009ml\u2009l\u20131 1\u2009M NH4Cl solution and 1.0\u2009ml\u2009l\u20131 1\u2009M sodium pyruvate solution (adjusted to pH\u20097.6 with NaOH).T.\u2009kodakarensis was cultured at 83\u2009\u00b0C, 87\u2009\u00b0C or 91\u2009\u00b0C, in nutrient-rich medium (ASW-YT-S0 or MA-YT-Pyr) or synthetic medium containing amino acids (ASW-AA-S0), under strict anaerobic conditions. ASW-YT-S0 medium contains 0.8\u00d7 artificial sea water (ASW)50, 10\u2009g\u2009l\u20131 yeast extract, 5.0\u2009g\u2009l\u20131 tryptone, 2.0\u2009g\u2009l\u20131 elemental sulfur and 0.1% (wt/vol) resazurin. MA-YT-Pyr medium contains 30.5\u2009g\u2009l\u20131 Marine Art SF-1 (Osaka Yakken), 10\u2009g\u2009l\u20131 yeast extract, 5.0\u2009g\u2009l\u20131 tryptone, 5.0\u2009g\u2009l\u20131 pyruvate sodium and 0.1% (wt/vol) resazurin. ASW-AA-S0 medium contains 0.8\u00d7 ASW, 0.5\u00d7 amino acid solution50, modified Wolfe\u2019s trace minerals 2, 0.01\u2009g\u2009l\u20131 H3BO3 and 0.01\u2009g\u2009l\u20131 NaMoO4\u00b72H2O), 5.0\u2009ml\u2009l\u20131 vitamin mixture51, 2.0\u2009g\u2009l\u20131 elemental sulfur and 0.1% (wt/vol) resazurin. For plate cultivation, 2.0\u2009ml\u2009l\u20131 polysulfide solution was added instead of elemental sulfur, and the media were solidified with 1.0% Gelrite . When pyrF-negative transformants were selected0, 75% 5-fluoroorotic acid (5-FOA) was added. We used ASW-YT-S0 medium for standard cultivation, MA-YT-Pyr medium for growth comparisons and ASW-AA-S0 medium for construction of the gene knockout strain.N-lauroylsarcosine sodium salt and 1\u2009mM 2-mercaptoethanol) and mixed with an equal volume of water-saturated phenol and 1/10 volume of 3\u2009M sodium acetate (pH\u20095.3). The mixture was shaken for 1\u2009h on ice and mixed with 1/5 volume of chloroform, followed by centrifugation at 8,000g for 10\u2009min at 4\u2009\u00b0C. The supernatant was collected and mixed with an equal volume of chloroform, followed by centrifugation at 8,000g for 10\u2009min at 4\u2009\u00b0C. Total RNA was obtained from the resultant supernatant by isopropanol precipitation. The total RNA prepared in this manner was separated by 10% denaturing PAGE, followed by staining with SYBR Gold or toluidine blue. The visualized tRNA fraction including class I and class II tRNAs was cut out and eluted from the gel slice with elution buffer (0.3\u2009M sodium acetate (pH\u20095.3) and 0.1% (wt/vol) SDS), followed by filtration to remove the gel pieces and ethanol precipitation for RNA-MS analysis of the tRNA fraction.For small-scale preparation (~100-ml culture), archaeal cells were resuspended in 3\u2009ml solution D , 0.5% (wt/vol) S.\u2009tokodaii, cell pellets (53\u2009g) were resuspended in 530\u2009ml solution D and then mixed with 53\u2009ml of 3\u2009M sodium acetate (pH\u20095.3) and 425\u2009ml neutralized phenol. The mixture was shaken for 1\u2009h on ice to which 106\u2009ml chloroform/isoamyl alcohol (49:1) was added, followed by centrifugation at 4,500g for 20\u2009min at 4\u2009\u00b0C. The supernatant was collected and mixed with 106\u2009ml chloroform/isoamyl alcohol (49:1), followed by centrifugation at 4,500g for 15\u2009min at 4\u2009\u00b0C. The aqueous phase was collected and then subjected to isopropanol precipitation. The collected RNA was resuspended in 53\u2009ml water and mixed with 80\u2009ml TriPure Isolation Reagent (Roche), followed by centrifugation at 10,000g for 20\u2009min at 4\u2009\u00b0C. The supernatant was collected and mixed with 36\u2009ml chloroform/isoamyl alcohol (49:1), followed by centrifugation at 10,000g for 10\u2009min at 4\u2009\u00b0C. The aqueous phase was collected and precipitated with isopropanol. The prepared total RNA (608\u2009mg) was dissolved in 250\u2009ml of buffer consisting of 20\u2009mM HEPES-KOH (pH\u20097.6), 200\u2009mM NaCl and 1\u2009mM DTT and then loaded on a DEAE Sepharose Fast Flow column (320-ml beads) and fractionated with a gradient of NaCl from 200 to 500\u2009mM. Fractions containing tRNA were collected by isopropanol precipitation.For large-scale preparation of tRNA fractions from 24 or chaplet column chromatography (CCC)52. Approximately 200 absorbance at 260\u2009nm (A260) units of the S.\u2009tokodaii tRNA fraction was subjected to RCC. The isolation procedure was carried out as follows: hybridization at 66\u2009\u00b0C in 6\u00d7 NHE buffer (30\u2009mM HEPES-KOH (pH\u20097.5), 15\u2009mM EDTA (pH\u20098.0), 1.2\u2009M NaCl, 1\u2009mM DTT), washing at 50\u2009\u00b0C with 0.1\u00d7 NHE buffer (0.5\u2009mM HEPES-KOH (pH\u20097.5), 0.25\u2009mM EDTA (pH\u20098.0), 20\u2009mM NaCl, 0.5\u2009mM DTT) and elution at 72\u2009\u00b0C with 0.1\u00d7 NHE buffer. Eluted tRNAs were recovered by ethanol precipitation. Mature and precursor tRNAs were separated by 10% denaturing PAGE and stained with SYBR Gold. Visualized bands of mature and precursor tRNAs were cut out and eluted from the gel slices with elution buffer, followed by filtration to remove the gel pieces and precipitation with ethanol.Isolation of individual tRNAs from thermophilic organisms is extremely difficult owing to their high melting temperatures, which are the consequence of their high G+C content and complex modifications. We thus optimized our original method for RNA isolation by RCCp47, we conducted large-scale isolation of S.\u2009tokodaii tRNAVal3 using CCC52. The S.\u2009tokodaii tRNA fraction was subjected to CCC with tandem affinity chaplet columns for tRNAVal3, tRNAIle2 and tRNAPhe. The isolation procedure was carried out as follows: hybridization at 66\u2009\u00b0C in 6\u00d7 NHE buffer, washing separately at 50\u2009\u00b0C with 0.1\u00d7 NHE buffer and elution at 72\u2009\u00b0C with 0.1\u00d7 NHE buffer. The eluted tRNAs were recovered by isopropanol precipitation. The sequences of the DNA probes are shown in Supplementary Table Val3 was further purified by anion exchange chromatography to completely remove tRNAVal2, as described below.To crystalize native tRNA bearing U 2,000\u2009A20 units w1 (Epicentre or Thermo Fisher Scientific) or RNase\u2009A (Ambion) and analysed with a linear ion trap\u2013Orbitrap hybrid mass spectrometer equipped with a custom-made nanospray ion source and a splitless nanoHPLC system as described previously27. To analyse \u03a8 sites, tRNA was treated with acrylonitrile to cyanoethylate \u03a853 and subjected to RNA-MS. For dephosphorylation of the Up47-containing fragment . To precisely map tRNA modifications, RNA fragments were decomposed by CID in the instrument. The normalized collision energy of LTQ Orbitrap XL was set to 40%. Mongo Oligo Mass Calculator v2.08 was used for assignment of the product ions in CID spectra.For tRNA fragment analysis by RNA-MS, 30\u2009ng (900\u2009fmol) of the isolated tRNA or 150\u2009ng (4.5\u2009pmol) of tRNA mixture was digested with RNase\u2009TVal3 was digested with 0.09\u2009U nuclease\u2009P1 in 20\u2009mM ammonium acetate (pH\u20095.2) at 50\u2009\u00b0C for 1\u2009h and mixed with 1/8 volume of 1\u2009M trimethylamine-HCl (TMA-HCl) (pH\u20097.2) and 0.06\u2009U phosphodiesterase\u2009I , followed by incubation at 37\u2009\u00b0C for 1\u2009h. To this mixture, 0.08\u2009U BAP was added, and the sample was incubated at 50\u2009\u00b0C for 1\u2009h. After that, 9 volumes of acetonitrile were added, followed by LC\u2013MS/MS analysis as described in refs. 54 with some modifications as follows. The samples were chromatographed with a ZIC-cHILIC column and eluted with 5\u2009mM ammonium acetate (pH\u20095.3) (solvent\u2009A) and acetonitrile (solvent\u2009B) at a flow rate of 100\u2009\u03bcl\u2009min\u20131 with a multistep linear gradient: 90\u201350% solvent\u2009B for 30\u2009min, 50% solvent\u2009B for 10\u2009min, 50\u201390% solvent\u2009B for 5\u2009min and then initialization with 90% solvent\u2009B. The chromatographed eluent was directly introduced into the electrospray ionization source of the Q Exactive Hybrid Quadrupole\u2013Orbitrap mass spectrometer (Thermo Fisher Scientific).For nucleoside analysis, 800\u2009ng (24\u2009pmol) of the isolated tRNA1 in 20\u2009mM ammonium acetate (pH\u20095.2) at 50\u2009\u00b0C for 1\u2009h and then mixed with 9 volumes of acetonitrile for LC\u2013MS. The digests were chromatographed with a ZIC-cHILIC column and analysed by Q Exactive Hybrid Quadrupole\u2013Orbitrap mass spectrometer (Thermo Fisher Scientific) or LTQ Orbitrap XL (Thermo Fisher Scientific) with a multistep linear gradient: 90\u201350% solvent\u2009B for 30\u2009min, 50% solvent\u2009B for 10\u2009min, 50\u201390% solvent\u2009B for 5\u2009min and then initialization with 90% solvent\u2009B.For nucleotide analysis, 800\u2009ng (24\u2009pmol) of the tRNA fraction or individual tRNA was digested with 0.09\u2009U nuclease\u2009PThe acquired LC\u2013MS data were analysed using Xcalibur 4.1 (Thermo Fisher Scientific) and were visualized with Canvas X .A260 units of the S.\u2009tokodaii tRNA fraction was completely digested with nuclease\u2009P1. Digests containing pN324m5C dinucleotide were subjected to periodate oxidation with 10\u2009mM NaIO4 for 1\u2009h on ice in the dark. The reaction was stopped by addition of 1\u2009M l-rhamnose and incubation for 30\u2009min. For \u03b2-elimination, an equal volume of 2\u2009M lysine-HCl (pH\u20098.5) was added, and the sample was incubated at 45\u2009\u00b0C for 90\u2009min. The product containing pN324p was then subjected to anion exchange chromatography with a Q Sepharose Fast Flow column equilibrated with 20\u2009mM triethylammonium bicarbonate (TEAB) (pH\u20098.2). The eluate with 2\u2009M TEAB was collected and dried by evaporation in vacuo. The pellet was dissolved with water and mixed with an equal volume of chloroform, followed by centrifugation at 20,000g for 5\u2009min at 4\u2009\u00b0C. The supernatant was recovered and dried again. This process was repeated five times. The resultant digest was mixed with 9 volumes of acetonitrile and subjected to LC\u2013MS/MS using an LCQ-Advantage ion trap mass spectrometer (Thermo Scientific), equipped with an electrospray ionization source and an HP1100 LC system (Agilent Technologies). For LC, the digest was chromatographed with a ZIC-HILIC column and eluted with 5\u2009mM formic acid (pH\u20093.4) (solvent\u2009A) and acetonitrile (solvent\u2009B) at a flow rate of 100\u2009\u03bcl\u2009min\u20131 with a multistep gradient: 90\u201370% solvent\u2009B for 25\u2009min, 70\u201310% solvent\u2009B for 15\u2009min, 10% solvent\u2009B for 5\u2009min and then initialized with 90% solvent\u2009B.Five arkI from T.\u2009kodakarensis, Methanocaldococcus fervens, P.\u2009oguniense, Aquifex aeolicus, Nautilia profundicola and Leptolyngbya sp. PCC7376 were designed with codons optimized for E.\u2009coli expression and synthesized by GENEWIZ or Thermo Fisher Scientific. Each gene was cloned into the pE-SUMO-TEV vector by the SLiCE method55. N.\u2009viennensis arkI was PCR amplified from genomic DNA with a set of primers (Supplementary Table Synthetic genes for E.\u2009coli BL21(DE3) or Rosetta2(DE3) cells transformed with the pE-SUMO-TEV vector carrying each arkI gene were cultured in 250\u2009ml or 1\u2009l of LB containing 50\u2009\u03bcg\u2009ml\u20131 kanamycin and 20\u2009\u03bcg\u2009ml\u20131 chloramphenicol when necessary. His6\u2013SUMO-tagged recombinant protein was expressed at 37\u2009\u00b0C for 3\u20134\u2009h by induction with 0.1 or 1\u2009mM IPTG or 2% (wt/vol) lactose when the cells reached OD610\u2009=\u20090.4\u20130.6. P.\u2009oguniense ArkI was expressed in cells cultured overnight at 18\u2009\u00b0C. The collected cells were resuspended in lysis buffer (50\u2009mM HEPES-KOH (pH\u20098.0), 150\u2009mM KCl, 2\u2009mM MgCl2, 20\u2009mM imidazole, 12% (vol/vol) glycerol, 1\u2009mM 2-mercaptoethanol and 1\u2009mM PMSF) and disrupted by sonication, followed by centrifugation at 15,000g for 15\u2009min at 4\u2009\u00b0C. The supernatant was boiled at 60\u2009\u00b0C for 20\u2009min and centrifuged at 15,000g for 15\u2009min at 4\u2009\u00b0C. The recombinant protein was affinity captured on an Ni-Sepharose 6 Fast Flow column and then eluted with lysis buffer containing 300\u2009mM imidazole, followed by gel filtration with a PD-10 column to remove the imidazole. The recombinant protein for N.\u2009viennensis ArkI was purified using a HisTrap column with a linear gradient of 0\u2013500\u2009mM imidazole, followed by dialysis using a Slide-A-Lyzer Dialysis Cassette (Thermo Fisher Scientific) to remove imidazole. The purified protein was subjected to Ulp1 digestion at 4\u2009\u00b0C overnight to cleave the His6\u2013SUMO tag and then passed through a Ni-Sepharose 6 Fast Flow column to remove the tag. Because ArkI homologues from M.\u2009fervens (MfArkI) and Leptolyngbya sp. PCC7376 (LeArkI) aggregated following tag removal, His6\u2013SUMO tag-fused proteins of these homologues were used for the phosphorylation assay. Purified protein was quantified by the Bradford method using BSA as a standard.T.\u2009kodakarensis ArkI for crystallization, the E.\u2009coli BL21(DE3) strain carrying pE-SUMO-TkArkI was cultured in 2\u2009l of LB containing 50 \u03bcg\u2009ml\u20131 kanamycin and TkArkI was expressed at 25\u2009\u00b0C overnight by induction with 0.1\u2009mM IPTG when the cells reached OD610\u2009=\u20090.4. The cells were collected and disrupted by sonication in lysis buffer (50\u2009mM HEPES-KOH (pH\u20098.0), 150\u2009mM KCl, 2\u2009mM MgCl2, 20\u2009mM imidazole, 12% (vol/vol) glycerol, 1\u2009mM 2-mercaptoethanol and 1\u2009mM PMSF). The protein was purified using a HisTrap column with a linear gradient of 20\u2013520\u2009mM imidazole. Fractions containing TkArkI were pooled and subjected to Ulp1 digestion at 4\u2009\u00b0C overnight to cleave the tag, followed by passage through a Ni-Sepharose 6 Fast Flow column to remove the tag fragment. The flow-through fraction was filtered through a 0.45-\u03bcm PVDF membrane to remove the resin. The protein was further purified by affinity chromatography with a HiTrap Heparin HP column using a linear gradient of 150\u20131,150\u2009mM KCl. TkArkI was further purified by size exclusion chromatography using a Superdex 75 10/300 GL column with buffer containing 20\u2009mM Tris-HCl (pH\u20098.0), 150\u2009mM NaCl and 10\u2009mM 2-mercaptoethanol and then concentrated to 5.74\u2009mg\u2009ml\u20131 and stored at \u201380\u2009\u00b0C.For large-scale preparation of T.\u2009kodakarensis kptA gene was PCR amplified from genomic DNA from T.\u2009kodakarensis with the primers listed in Supplementary Table tkkptA. The E.\u2009coli Rosetta2(DE3) strain carrying pE-SUMO-TEV-tkkptA was cultured in 1\u2009l LB containing 50\u2009\u03bcg\u2009ml\u20131 kanamycin and 20\u2009\u03bcg\u2009ml\u20131 chloramphenicol, and TkKptA was expressed at 37\u2009\u00b0C for 3\u2009h by induction with 0.1\u2009mM IPTG when the cells reached OD610\u2009=\u20090.6. The recombinant TkKptA was purified as described above. The gene encoding Tpt1p was PCR amplified from the genomic DNA of S.\u2009cerevisiae BY4742 with the set of primers listed in Supplementary Table The p47 by yeast Tpt1p was performed as described33. Individual tRNAs or the tRNA fraction was incubated for 3\u2009h at 30\u2009\u00b0C in a reaction mixture (25\u2009\u03bcl) consisting of 20\u2009mM Tris-HCl (pH\u20097.4), 0.5\u2009mM EDTA (pH\u20098.0), 1\u2009mM NAD+, 2.5\u2009mM spermidine, 0.1\u2009mM DTT, 0.9\u2009\u03bcM tRNA and 0.1\u2009\u03bcg\u2009\u03bcl\u20131 recombinant Tpt1p. The tRNA was extracted by phenol/chloroform treatment and recovered by ethanol precipitation, followed by desalting with Centri-Sep spin columns (Princeton Separations). For crystallization of Tpt1p-treated tRNA, S.\u2009tokodaii tRNAVal3 (202.5\u2009\u03bcg) was dephosphorylated by yeast Tpt1p in a 200-\u03bcl reaction mixture.Removal of the 2\u2032-phosphate of US.\u2009tokodaii tRNAVal3 (25\u2009pmol) with or without Up47 was dissolved in degassed buffer consisting of 50\u2009mM Tris-HCl (pH\u20097.4), 100\u2009mM NaCl and 1\u2009mM MgCl2 and incubated at 80\u2009\u00b0C for 5\u2009min, followed by cooling to 25\u2009\u00b0C at a rate of 0.1\u2009\u00b0C\u2009s\u20131. The samples were placed onto a Type 8 multi-micro UV quartz cell . The hyperchromicity of tRNA was monitored on a UV\u2013visible light spectrophotometer . The gradients were as follows: 25\u2009\u00b0C for 30\u2009s, 25\u201340\u2009\u00b0C at 5\u2009\u00b0C\u2009min\u20131, 40\u2009\u00b0C for 5\u2009min and 40\u2013105\u2009\u00b0C at 0.5\u2009\u00b0C\u2009min\u20131. The Tm was calculated using Spectra Manager v2 (JASCO). Melting curves were generated using Microsoft Excel.S.\u2009tokodaii tRNAVal3 (25\u2009pmol) with or without Up47 was labelled with 32P at the 3\u2032\u2009terminus by ligation with [5\u2032-32P]cytidine 3\u2032,5\u2032-bisphosphate (PerkinElmer). The labelled tRNA was separated on a 7.5% (wt/vol) polyacrylamide gel containing 7\u2009M urea, 1\u00d7 TBE and 10% (vol/vol) glycerol and was purified by gel extraction. Labelled tRNA was mixed with the S.\u2009tokodaii tRNA fraction as a carrier to a concentration of 100,000\u2009counts per minute (c.p.m.) per A260 unit and was precipitated with ethanol. The pellet was dissolved in water to a concentration of 0.1\u2009A260 units per \u03bcl. For the RNase degradation assay, the labelled tRNA was incubated at 65\u2009\u00b0C in a reaction mixture consisting of 10\u2009mM HEPES-KOH (pH\u20097.6), 0.5\u2009mM MgCl2, 100\u2009mM NaCl and 0.1\u2009U\u2009\u03bcl\u20131 RNase\u2009I (Promega). At time points of 1, 3, 5, 10, 15 and 30\u2009min after starting the reaction, aliquots were taken from the mixture and mixed well with chilled phenol/chloroform/isoamyl alcohol to stop the reaction, followed by centrifugation at 15,000g for 15 min at 4\u2009\u00b0C. The supernatant was collected and treated with an equal volume of chloroform, followed by centrifugation at 15,000g for 5\u2009min at 4\u2009\u00b0C. The supernatant was mixed with 2\u00d7 loading solution sucrose, 0.05% (wt/vol) xylene cyanol and 0.05% (wt/vol) bromophenol blue) and subjected to 10% denaturing PAGE. The gel was exposed to an imaging plate, and radioactivity was visualized by using an FLA-7000 imaging analyser (Fujifilm). Graphs were generated using Microsoft Excel.S.\u2009tokodaii tRNAVal3 (500\u2009\u03bcg), isolated as described above, was refolded in annealing buffer (50\u2009mM HEPES-KOH (pH\u20097.6), 5\u2009mM MgCl2 and 1\u2009mM DTT) by incubation for 5\u2009min at 80\u2009\u00b0C and cooling to 25\u2009\u00b0C with a rate of 0.1\u2009\u00b0C\u2009s\u20131. tRNAVal3 was further purified by anion exchange chromatography using a Mono Q 5/50 GL column with a linear gradient of 200\u20131,000\u2009mM NaCl. The major peak was collected, precipitated with isopropanol, dissolved in water and precipitated with ethanol. Tpt1p-treated tRNAVal3 was prepared with the same procedure as described above. The purified tRNA was dissolved in buffer consisting of 10\u2009mM Tris-HCl (pH\u20097.1) and 5\u2009mM MgCl2 to a concentration of 50\u2009\u03bcM. One microlitre of tRNA solution was mixed with 1\u2009\u03bcl Natrix 2 no. 32 and 30% (vol/vol) MPD) (Hampton Research) on silicon-coated glass and crystalized by the hanging drop vapor diffusion method at 20\u2009\u00b0C.\u20131 before crystallization. One microlitre of the protein solution was mixed with 0.5\u2009\u03bcl reservoir solution, containing 25% (vol/vol) ethylene glycol. TkArkI was crystallized by the hanging drop vapor diffusion method at 20\u2009\u00b0C.The concentration of TkArkI was adjusted to 5\u2009mg\u2009mlVal3 crystals, the crystals were cryoprotected with a portion of the reservoir solution. For data collection for the native TkArkI crystal, the crystal was cryoprotected with solution containing 25% (vol/vol) ethylene glycol, 2\u2009mM MgCl2 and 1\u2009mM ATP. For data collection for the iodide-derivative TkArkI crystal, the crystal was briefly soaked in and cryoprotected with solution containing 300\u2009mM potassium iodide and 22.5% (vol/vol) ethylene glycol, and the diffraction dataset was collected at a wavelength of 1.5\u2009\u00c5. The datasets were indexed, integrated and scaled using xds56. The initial phase of tRNAVal3 was determined by molecular replacement with Phaser57. The structure of T.\u2009thermophilus tRNAVal 58 was used for the model. The initial phase of TkArkI was determined by the SAD method using the anomalous signal of iodide ions. The iodine sites were located by SHELX59, and the initial phase was calculated by Phaser. Subsequent density modification and initial model building were performed with RESOLVE60. The model was further modified with Coot61 and refined with Phenix62. Crystal structures and their electron density maps were visualized using PyMOL, Cuemol or Coot. Torsion angles of the tRNAs were analysed with DSSR software63.The datasets were collected at beamline BL-17A at the Photon Factory at KEK, Japan. For data collection for the tRNA15N]adenosine (10\u2009pmol) and [15N]guanosine (10\u2009pmol) as tracer molecules, followed by addition of 4 volumes of methanol, an equal volume of chloroform and 3 volumes of water and vigorous mixing. The denatured protein was removed by centrifugation at 15,000g for 1\u2009min at 4\u2009\u00b0C. The supernatant was dried in vacuo and dissolved in 20\u2009\u03bcl water. Half of the extract was analysed by LC\u2013MS. The tracer molecules were prepared by dephosphorylation of [15N]ATP and [15N]GTP as follows: 1,000\u2009pmol each of [15N]ATP (Silantes) and [15N]GTP (Silantes) was treated with 0.04\u2009U alkaline phosphatase in 20\u2009mM ammonium acetate (pH\u20098.0) at 60\u2009\u00b0C for 30\u2009min. After dephosphorylation, PAP was heat denatured at 95\u2009\u00b0C for 5\u2009min.TkArkI purified by affinity chromatography with a HiTrap Heparin HP column (100\u2009pmol) was mixed with ATP to tRNA similarly to the kinetic studies of TkArkI (see below). tRNA phosphorylation was performed at 70\u2009\u00b0C for 15\u2009min in an 8-\u03bcl reaction mixture. For PAGE analysis, 4\u2009\u03bcl of the reaction mixture was mixed with 4\u2009\u03bcl of 2\u00d7 loading solution, resolved by 10% denaturing PAGE and exposed to an imaging plate to visualize radiolabelled RNA with an FLA-9000 imaging analyser (Fujifilm). The gel image was analysed using Multi Gauge (Fujifilm). Bar graphs with independent plots were prepared with R (R Foundation). For phosphorylation of total RNA, the reaction was performed at 70\u2009\u00b0C for 30\u2009min in an 8-\u03bcl reaction mixture consisting of 50\u2009mM HEPES-NaOH (pH\u20097.5), 1\u2009mM MgCl2, 1\u2009mM MnCl2, 1\u2009mM DTT, 10% (vol/vol) glycerol, 100\u2009\u03bcM [\u03b3-32P]ATP , 1.8\u2009\u03bcM TkArkI and 50\u2009ng\u2009\u03bcl\u20131 total RNA fraction (from the T.\u2009kodakarensis \u0394arkI strain). Then, 0.5\u2009\u03bcl of 50\u2009mM EDTA (pH\u20098.0) was added, and 4\u2009\u03bcl of reaction mixture was mixed with 2\u00d7 loading solution, resolved by 10% denaturing PAGE and visualized as described above.Up47 by other ArkI homologues was carried out at 70\u2009\u00b0C for 30\u2009min in a reaction mixture (30\u2009\u03bcl) containing 50\u2009mM PIPES-NaOH (pH\u20096.9), 125\u2009mM NaCl, 1\u2009mM MgCl2, 1\u2009mM MnCl2, 1\u2009mM DTT, 10% (vol/vol) glycerol, 500\u2009\u03bcM ATP, 0.05\u2009mg\u2009ml\u20131 BSA (Takara), 1\u2009\u03bcM tRNA transcript and 0.5\u2009\u03bcM ArkI protein. For NvArkI, the reaction temperature was set to 45\u2009\u00b0C. For ArkI homologue from N.\u00a0profundicola\u00a0(NpArkI), the reaction was carried out at 50\u2009\u00b0C for 60\u2009min. After the reaction, tRNA was prepared as described above. For PAGE analysis, Up47 formation was carried out in a reaction mixture (8\u2009\u03bcl) containing 50\u2009mM PIPES-NaOH (pH\u20096.9), 125\u2009mM NaCl, 1\u2009mM MgCl2, 1\u2009mM MnCl2, 1\u2009mM DTT, 10% (vol/vol) glycerol, 100\u2009\u03bcM [\u03b3-32P]ATP , 0.1\u2009mg\u2009ml\u20131 BSA (Takara), 0.75\u2009\u03bcM recombinant ArkI homologue and 50\u2009ng\u2009\u03bcl\u20131 E.\u2009coli total RNA. Then, the reaction mixture was mixed with 2\u00d7 loading solution, resolved by 10% denaturing PAGE and visualized as described above.Formation of Up47 by TkKptA was carried out at 60\u2009\u00b0C for 1\u2009h in a reaction mixture (30\u2009\u03bcl) containing 20\u2009mM Tris-HCl (pH\u20097.4), 0.5\u2009mM EDTA (pH\u20098.0), 1\u2009mM NAD+, 2.5\u2009mM spermidine, 0.1\u2009mM DTT, 0.9\u2009\u03bcM T.\u2009kodakarensis tRNA fraction and 0.1\u2009\u03bcg\u2009\u03bcl\u20131 recombinant TkKptA. After the reaction, the tRNA was extracted by acidic phenol/chloroform, desalted on a NAP-5 column and precipitated with isopropanol. For RNA-MS, the prepared tRNA was desalted by drop dialysis as described above.Dephosphorylation of Up47 formation was quantified by \u03b3-phosphate transfer from [\u03b3-32P]ATP to tRNA. For kinetic measurement of the tRNA substrate, tRNA phosphorylation was performed at 70\u2009\u00b0C in a reaction mixture (25\u2009\u03bcl) consisting of 50\u2009mM PIPES-NaOH (pH\u20096.9), 125\u2009mM NaCl, 1\u2009mM MgCl2, 1\u2009mM MnCl2, 1\u2009mM DTT, 10% (vol/vol) glycerol, 100\u2009\u03bcM [\u03b3-32P]ATP , 0.05\u2009mg\u2009ml\u20131 BSA (Takara), 0.05\u2009\u03bcM TkArkI and 0.1\u20135.0\u2009\u03bcM of in vitro-transcribed T.\u2009kodakarensis tRNAVal3. For kinetic measurement of the ATP substrate, the ATP concentration was altered from 15.6 to 1,000\u2009\u03bcM [\u03b3-32P]ATP and the tRNA concentration was increased to 1.0\u2009\u03bcM. At each time point (2 and 5\u2009min), 8-\u03bcl aliquots were taken and mixed with an equal volume of 2\u00d7 loading solution bromophenol blue, 0.2% (wt/vol) xylene cyanol and 50\u2009mM EDTA (pH\u20098.0)) to quench the reaction. Each sample was subjected to 10% denaturing PAGE. The gel was exposed on an imaging plate to measure radiolabelled tRNAs using an FLA-9000 imaging analyser. Kinetic parameters were calculated using Prism 7 (GraphPad).TkArkI-mediated Up47 was quantified by measuring the reduction in radioactivity for tRNA. In vitro-transcribed T.\u2009kodakarensis tRNAVal3 was phosphorylated by TkArkI with [\u03b3-32P]ATP as described above and then purified by gel extraction and isopropanol precipitation. In addition, the same tRNA was phosphorylated by TkArkI with unlabelled ATP. By mixing labelled and unlabelled tRNAs, the specific activity of the labelled tRNA was adjusted to 6,250\u2009c.p.m. per pmol in buffer consisting of 50\u2009mM HEPES-KOH (pH\u20097.6), 5\u2009mM MgCl2 and 1\u2009mM DTT. The labelled tRNA was incubated at 80\u2009\u00b0C for 5\u2009min and then cooled at room temperature, followed by isopropanol precipitation. The labelled tRNA was dissolved in water to a concentration of 8\u2009\u03bcM . Dephosphorylation of the labelled tRNA by TkKptA was performed at 70\u2009\u00b0C in a reaction mixture (30\u2009\u03bcl) consisting of 50\u2009mM PIPES-NaOH (pH\u20096.9), 125\u2009mM NaCl, 1\u2009mM MgCl2, 1\u2009mM MnCl2, 1\u2009mM DTT, 10% (vol/vol) glycerol, 1\u2009mM NAD+, 0.05\u2009mg\u2009ml\u20131 BSA (Takara), 1\u2009nM TkKptA and 12.5\u2013800\u2009nM 32P-labelled tRNA. At each time point (2 and 5\u2009min), 8-\u03bcl aliquots were spotted on Whatman 3MM filter paper, which was immediately soaked in 5% (wt/vol) trichloroacetic acid. The filter paper was washed three times for 15 min with ice-cold 5% (wt/vol) trichloroacetic acid, rinsed for 5\u2009min with ice-cold ethanol and dried in air. Radioactivity on the filter paper was measured by liquid scintillation counting . Kinetic parameters were calculated using Prism 7.TkKptA-mediated dephosphorylation of UN.\u2009viennensis arkI was PCR amplified and cloned into pMW118 (Invitrogen) under the control of the synthetic constitutive J23106 promoter68, followed by insertion of sequences encoding a His6 tag and a 3\u00d7Flag tag at the C terminus of the N.\u2009viennensis arkI gene, yielding pMW-J23106-nvarkI . The ampicillin resistance cassette (Ampr) was replaced with a chloramphenicol resistance cassette (Camr), yielding pQE-80LC-tkkptA, pQE-80LC-eckptA and pQE-80LC-sctpt1, respectively (Supplementary Table E.\u2009coli \u0394trmB\u0394tapT (Kanr) strain was transformed with pMW-J23106-nvarkI and further transformed with pQE-80LC-tkkptA, pQE-80LC-eckptA or pQE-80LC-sctpt1. The transformants were inoculated in 3\u2009ml LB supplemented with 20\u2009\u03bcg\u2009ml\u20131 chloramphenicol, 50\u2009\u03bcg\u2009ml\u20131 kanamycin and 100\u2009\u03bcg\u2009ml\u20131 ampicillin and cultured at 37\u2009\u00b0C until mid-log phase. When the OD610 reached 0.6, IPTG was added to a final concentration of 10 or 100\u2009\u03bcM to induce expression of the KptA/Tpt1p homologue and cells were cultured for 3.5\u2009h. A 1.5-ml aliquot of the culture was taken, and the tRNA fraction was extracted and analysed by shotgun RNA-MS as described above. Primers, E.\u2009coli strains and plasmids used are listed in Supplementary Tables Chemical structures were drawn with chemical structure drawing tools, including ACD/ChemSketch (ACD/Labs) or ChemDraw (PerkinElmer).Further information on research design is available in the\u00a0Any methods, additional references, Nature Research reporting summaries, source data, extended data, supplementary information, acknowledgements, peer review information; details of author contributions and competing interests; and statements of data and code availability are available at 10.1038/s41586-022-04677-2.Supplementary InformationThis file contains Supplementary Notes 1\u20137, Supplementary Figs. 1\u201310, Supplementary Tables 1\u20137 and the legend for Supplementary Video 1.Reporting SummaryPeer Review FileSupplementary Video 1S. tokodaii tRNAVal3.Structural switch of the core region in"} +{"text": "The crystal structure of 1 is similar to that of the bromide salt (2), possessing anion\u00b7\u00b7\u00b7C5F5N\u00b7\u00b7\u00b7C6H5 motifs, whilst that of 3 contains columns of alternating iodide anions and parallel tetrafluoropyridyl rings. All three crystal structures possess C(1)\u2013H\u2219\u2219\u2219X\u2212 and C(2)\u2013H\u2219\u2219\u2219X\u2212 hydrogen bonding. DFT calculations reveal that the strengths of the hydrogen bonding interactions lie in the order C(1)\u2013H\u00b7\u00b7\u00b7X\u2212 > C(3)\u2013H\u00b7\u00b7\u00b7X\u2212 > C(2)\u2013H\u00b7\u00b7\u00b7X\u2212 for the same halide (X\u2212) and Cl\u2212 > Br\u2212 > I\u2212 for each position. It is suggested that salt 3 adopts a different structure to salts 1 and 2 because of the larger size of iodide.The crystal structures of 1--3-benzylimidazolium chloride ( However, we hypothesize that simple monoatomic anions, which differ in size and consequent properties, such as polarizability, can also exert a strong influence on the crystal structures adopted by 1-polyfluoroaryl-3-benzylimidazolium salts. In order to test this hypothesis we chose to investigate the crystal structures of the halide salts of 1--3-benzylimidazolium and polively + and [(NC5F4NC3H3NCH2Ph).2X]\u2212 peaks in the positive and negative ion mass spectra, respectively of 1 (X = Cl) and 3 (X = I), and the absence of [(NC5F4NC3H3NCH2Ph)2.Br]+ and [(NC5F4NC3H3NCH2Ph).2Br]\u2212. Attempts to prepare the fluoride analogue by the same method were unsuccessful. Salt 1 crystallized from chloroform in the polar orthorhombic space group Pna21, the same as that of 2. Salt 3 crystallized from dichloromethane in the centrosymmetric monoclinic space group P21/c. Crystal data are given in 1 and 3, together with the positions of the closest halide anions, are shown in 1--3-benzylimidazolium chloride \u2013C(9) for 1 and 2, but parallel for 3 \u2013H\u00b7\u00b7\u00b7X\u2212 hydrogen bonding, with that of salt 3 possessing two C(1)\u2013H\u00b7\u00b7\u00b7X\u2212 interactions. As expected, the C\u00b7\u00b7\u00b7X\u2212 distances \u2212 378.0516. C30H20F8N635Cl3 requires 723.0784; found [2M + Cl]\u2212 723.1200.MS (positive ion): C3.\u00a0\u00a015H10F4N3 requires 308.0811; found [M \u2212 I]+ 308.1033. C30H20F8N6I requires 743.0666; found [2M \u2212 I]+ 743.1140. MS (negative ion): I requires 126.9045; found [M \u2212 C15H10F4N3]\u2212 126.9181. C15H10F4N3I2 requires 561.8900; found [M + I]\u2212 561.9393.MS (positive ion): C1 and 3 were obtained by slow evaporation of solvent from solutions in chloroform and dichloromethane, respectively. Crystal data are listed in \u03b1 radiation. The structures of 1 and 3 were solved using Olex2 [3)) was used for subsequent refinements. The function minimized was [w(|Fo|2 \u2212 |Fc|2)] with reflection weights w\u22121 = [\u03c32 |Fo|2 + (g1P)2 + (g2P)] where p = [max |Fo|2 + 2|Fc|2]/3. Crystals of ng Olex2 and refing Olex2 refineme1) and 2063364 (3) contain the supplementary crystallographic data for this paper. These data can be obtained free of charge from The Cambridge Crystallographic Data Centre via www.ccdc.cam.ac.uk/data_request/cif.CCDC 2063365 -3-benzylimidazolium bromide are 1.083 \u00c5 within experimental error [ 1.322 \u00c5 .\u2212\u00b7\u00b7\u00b7C5F4N\u00b7\u00b7\u00b7C6H5 motifs, whilst the iodide salt, because of the larger size of the anion, adopts a different crystal structure containing columns of alternating tetrafluoropyridyl rings and iodide anions. The strength of charge-assisted hydrogen bonding interactions lie in the order C(1)\u2013H\u00b7\u00b7\u00b7X\u2212 > C(3)\u2013H\u00b7\u00b7\u00b7X\u2212 > C(2)\u2013H\u00b7\u00b7\u00b7X\u2212 for the same anion and Cl\u2212 > Br\u2212 > I\u2212 for each position. The strengths of the X\u2212\u00b7\u00b7\u00b7C5F4N interactions also decreases in the order Cl\u2212 > Br\u2212 > I\u2212.1--3-benzylimidazolium chloride and bromide adopt similar crystal structures with X"} +{"text": "Correction to: BMC Biotechnol (2022) 22:33 10.1186/s12896-022-00765-3Following publication of the original article , the autIncorrect 22ama121022j0001\u2009\u2192\u2009Correct JP22ama121022j0001Incorrect 17H06305\u2009\u2192\u2009Correct JP17H06305Incorrect 20H03243\u2009\u2192\u2009Correct JP20H03243The original article has been corrected."} +{"text": "II ion has an N4 square-planar coordination geometry defined by the four N atoms of two bidentate 2,3-di-2-pyridyl\u00adpyrazine ligands.In the title complex, the central Pd 14H10N4)2](NO3)2\u00b7CH3CN, consists of a cationic PdII complex, two anions and one lattice solvent mol\u00adecule, all in general positions. In the complex, the PdII cation is four-coordinated in a slightly distorted square-planar geometry defined by the four N atoms of two bidentate 2,3-di-2-pyridyl\u00adpyrazine ligands. The complex, anions and solvent mol\u00adecule are linked by weak C\u2014H\u22efO inter\u00admolecular hydrogen bonds. In the crystal, the complex mol\u00adecules are stacked in columns along the a axis.The title compound, [Pd(C The Pd\u2014N bond lengths are almost equal [2.0170\u2005(18) to 2.0286\u2005(19)\u2005\u00c5]. In the crystal, the pyridine rings are considerably inclined to the least-squares plane of the [PdN4] unit [maximum deviation = 0.0204\u2005(7)\u2005\u00c5], with dihedral angles of 70.56\u2005(7) (ring N3\u22efC9), 67.63\u2005(6) (ring N4\u22efC14), 71.32\u2005(6) (ring N7\u22efC23) and 71.64\u2005(7)\u00b0 (ring N8\u22efC28). The nearly planar pyrazine rings [maximum deviation = 0.027\u2005(2)\u2005\u00c5] are fairly perpendicular to the [PdN4] unit plane, with dihedral angles of 82.07\u2005(7) (ring N1\u22efC4) and 84.20\u2005(7)\u00b0 (ring N5\u22efC18).The title compound consists of a cationic Pdle Fig.\u00a01. In the a axis. In the columns, numerous intra- and inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between adjacent six-membered rings are present. For Cg1 (the centroid of ring N4\u22efC14) and Cg1i [symmetry code: (i) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01), the centroid-to-centroid separation is 3.688\u2005(2)\u2005\u00c5 and the planes are parallel and shifted by 1.683\u2005\u00c5.In the crystal structure Fig.\u00a02, the com3)2\u00b72H2O and stirred for 1\u2005h at room temperature. The formed precipitate was recrystallized from MeOH/ether, washed with ether, and dried under vacuum, to give a white powder (0.378\u2005g). Crystals suitable for X-ray analysis were obtained by slow evaporation of a MeOH/CH3CN solution, at room temperature.To a solution of 2,3-di-2-pyridyl\u00adpyrazine in acetone (30\u2005ml) was added Pd(NO\u22123) and the deepest hole (\u22120.49\u2005e\u2005\u00c5\u22123) in the last difference-Fourier map are located 1.02 and 0.97\u2005\u00c5, respectively, from atom O3.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S241431462100153X/bh4059sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S241431462100153X/bh4059Isup2.hklStructure factors: contains datablock(s) I. DOI: 2062092CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of a biologically important pharmacophore containing chalcone is reported. 18H17FO5, the conformation about the C=C bond of the central enone group is trans. The dihedral angle between the benzene rings is 13.08\u2005(3)\u00b0. The hy\u00addroxy group attached to the benzene ring is involved in an intra\u00admolecular O\u2014H\u22efO hydrogen bond. In the crystal, weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into chains along [001].In the title mol\u00adecule, C Especially when they have a hydroxyl group at the ortho position of an aromatic ring adjacent to the carbonyl group, they play important roles as precursors to form other flavonoids such as flavones, flavanones, flavonols and isoflavones \u00b0. An intra\u00admolecular O5\u2014H5\u22efO1 hydrogen bond \u00b0] and the para meth\u00adoxy group is almost coplanar with the ring [C6\u2014C7\u2014O4\u2014C12 = 177.1\u2005(2)\u00b0]. The meth\u00adoxy group at the ortho position is rotated significantly from the ring plane [C4\u2014C5\u2014O2\u2014C10 = \u2212123.9\u2005(2)\u00b0]. In the crystal, weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into chains propagating along [001] ethanone in 40\u2005ml of anhydrous ethanol was added 2,3,4-tri\u00admeth\u00adoxy\u00adbenzaldehyde and the temperature was adjusted to around 275\u2013277\u2005K in an ice bath. To the cooled reaction mixture was added 3\u2005ml of 40% aqueous KOH solution and the reaction mixture was stirred at room temperature for 20\u2005h. After completion of the reaction (monitored by thin-layer chromatography), this mixture was poured into ice water (100\u2005ml) and the resulting solution acidified with 6 Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314620000711/lh4051sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314620000711/lh4051Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314620000711/lh4051Isup3.cmlSupporting information file. DOI: 1979111CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II complexes bearing a tridentate polypyridine ligand and N-coordinating thio\u00adcyanato ligands at the axial or equatorial position are compared.The crystal structures of two Ru viz. cis-aqua\u00ad(thio\u00adcyanato-\u03baN)(tri\u00adphen\u00adyl\u00adphosphine-\u03baP)ruthenium(II) hexa\u00adfluorido\u00adphosphate\u2013acetone\u2013water (1/0.5/1), [Ru(NCS)(C21H13N5)(C18H15P)(H2O)]PF6\u00b70.5C3H6O\u00b7H2O (I) and trans-bis\u00ad(pyridine-\u03baN)(thiocyanato-\u03baN)ruthenium(II) thio\u00adcyanate, [Ru(NCS)(C21H13N5)(C5H5N)2]NCS (II), with an N-coordinating thio\u00adcyanato group and a tridentate polypyridyl supporting ligand, are reported. The RuII atom in each of the cationic complexes adopts a distorted octa\u00adhedral coordination sphere, defined by an N atom of the thio\u00adcyanato ligand, three N atoms from the tridentate polypyridyl ligand, and an O and P atom in (I) or two pyridine-N atoms in (II) derived from monodentate ligands. The thio\u00adcyanato ligand in (I) coordinates in an axial manner to the {Ru-dnp} unit , whereas it coordinates in an equatorial manner in (II). In the crystal structure of compound (I), intra\u00admolecular C\u2014H\u22efO, C\u2014H\u22efN and O\u2014H\u22efN hydrogen bonds as well as \u03c0\u2013\u03c0 contacts are present, in addition to inter\u00admolecular C\u2014H\u22efF, C\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds. In the crystal structure of compound (II), intra\u00admolecular C\u2014H\u22efN hydrogen bonds are observed along with inter\u00admolecular C\u2014H\u22efN and C\u2014H\u22efS hydrogen bonds as well as a \u03c0\u2013\u03c0 inter\u00adaction.The mol\u00adecular and crystal structures of two ruthenium(II) complexes, Their crystal structures are reported and compared in this communication.During the current study, the reaction of precursors with different axially bound ligands with the NCS2.II atoms in (I)et al., 20133 or pyridine and the non-coordinating N atoms in dnp or the monodentate ligands [OH2 in (I)NCS in (II)]. In (I)Cg1\u22efCg2 = 3.640\u2005(4)\u2005\u00c5 and Cg3\u22efCg4 = 3.749\u2005(3)\u2005\u00c5 where Cg1, Cg2, Cg3, and Cg4 are the centroids of the N1/C1\u2013C5, C29\u2013C34, N3/C9\u2013C13, and C35\u2013C40 rings, respectively] are present, with a slippage of 1.2\u2005\u00c5 for Cg1\u22efCg2. It is inferred from these results that both \u03c0\u2013\u03c0 inter\u00adactions are not exactly cofacial. The slippage angle \u03b2 is 19.2\u00b0 for Cg1\u22efCg2 and 16.2\u00b0 for Cg3\u22efCg4.Figs. 1II complexes containing polypyridines have been determined structurally, but the N-atom coordination is overwhelmingly dominant. These complexes can be distinguished crystallographically by the Ru\u2014X\u2014C bond angle (X = N or S) through the coordinating atom. For example, the Ru\u2014S\u2014C bond angles (for S-ligating examples) are 104\u2013106\u00b0 II atoms in both compounds exhibit an N-coordination.As mentioned above, it is important to distinguish the coordination atom of the thio\u00adcyanato ligand because of its ambidentate coordination mode. Both S- and N-coordinated RuII atom and the nitro\u00adgen atom in (I)II\u2013NCS}+ moieties per formula unit. Apart from Coulombic forces, there are weak C\u2014H\u22efF hydrogen bonds between the complex cation and the PF6\u2212 anion n Table\u00a01 and the n Table\u00a01.X (X = N or S) hydrogen-bonding inter\u00adactions exist between the complex cation and the NCS\u2212 anion (Table\u00a02Cg5\u22efCg5i = 4.0093\u2005(15)\u2005\u00c5; Cg5 is the centroid of the N5/C17\u2013C21 ring; symmetry code: (i) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z] with a centroid slippage of 1.263\u2005\u00c5 for Cg5\u22efCg5i are present. The slippage angle \u03b2 is 18.4\u00b0 for Cg5\u22efCg5i. These inter\u00adactions lead to the formation of a three-dimensional network structure n Table\u00a02 along wire Fig.\u00a04.4.II complexes with both N-coordinating thio\u00adcyanato and tridentate terpyridine derivative ligands (tpyR) of the form [Ru(tpyR)(NCS)L2]n have been reported, as revealed by a search of the Cambridge Crystal Structure Database 2(H2O)](PF6)2 . Crystals suitable for use in X-ray diffraction (XRD) studies were grown by vapor diffusion of diethyl ether into an acetone solution of (I)CN at 2130\u2005cm\u22121.A methano\u00adlic solution (40\u2005ml) containing [Ru(dnp)(PPh2(H2O)](PF6)2 . The yield was 9\u2005mg (40%). Single crystals suitable for XRD studies were obtained by recrystallization from acetone. FTIR using a KBr pellet showed \u03bdCN at 2121 (ligand) and 2055\u2005cm\u22121 (counter-ion).For the synthesis of compound (II)6.Uiso(H) = 1.2Ueq(C). The acetone solvent mol\u00adecule in (I)Table\u00a0310.1107/S2056989022004443/wm5641sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989022004443/wm5641Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989022004443/wm5641IIsup3.hklStructure factors: contains datablock(s) II. DOI: 2168839, 2168838CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecule is planar (r.m.s. deviation = 0.0164\u2005\u00c5) and in the crystal, inversion-symmetric dimers are formed as a result of pairs of strong O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds. A brief comparison is made with structurally related compounds deposited in the CSD. In addition, the synthesis and some spectroscopic details are presented.The structure of 2-chloro-1-(3-hy\u00addroxy\u00adphen\u00adyl)ethanone, C All bond lengths and angles fall within the expected ranges for organic structures.The mol\u00adecule of I Fig.\u00a01 is plana3.O\u22efO2i and O1i\u2014H1Oi\u22efO2 , giving an i and C2i\u2014H2i\u22efO2 (Table\u00a01i = 2.22\u2005(3)\u2005\u00c5]. These inter\u00adactions are all illustrated in Fig.\u00a02ii [symmetry code: (ii) \u2212x\u00a0+\u00a0y\u00a0\u2212\u00a0z\u00a0+\u00a01 screw-related mol\u00adecules, which loosely connect the dimers into layers parallel to . QAJNAS was added dropwise to a stirred mixture of 3-hy\u00addroxy\u00adaceto\u00adphenone in 5\u2005ml of methanol and 10\u2005ml of ethyl acetate/di\u00adchloro\u00admethane at 293\u2013303\u2005K. After completion of the addition, it was allowed to return to RT with stirring for 1\u2005h. The reaction was monitored by TLC. Then the solvent was removed under reduced pressure by rotary evaporation to give the desired product in 95% yield. An overall reaction scheme is depicted in Fig.\u00a04Spectroscopic data: Infrared and NMR spectroscopic details are as follows.\u22121): 3400 , 2987 (C\u2014H stretching), 1694 (C=C stretching), 1789 , 832 .FTIR : 4.7 , 5.671 , 7.14 , 7.36\u20137.4 , 7.493\u20137.51 .6.sp2\u2014H) and 0.99\u2005\u00c5 (R2CH2). The hydroxyl hydrogen atom coordinates were refined freely. In all cases, Uiso(H) values were set to 1.2Ueq of the attached atom.Crystal data, data collection, and structure refinement details are given in Table\u00a0210.1107/S2056989022009835/vm2272sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989022009835/vm2272Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022009835/vm2272Isup3.cmlSupporting information file. DOI: 2211527CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-[1-(2-pyrazin\u00adyl)ethyl\u00adidene]propane\u00adhydrazide, is a ligand able to form polynuclear metal complexes. The mol\u00adecule is not planar due to a twist between the oxime and amide groups. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds into supra\u00admolecular chains.The title compound, 2-hy\u00addroxy\u00adimino- 9H11N5O2, the oxime and hydrazide groups are situated in a cis-position in relation to the C\u2014C bond linking the two functional groups. The CH3C(=NOH)C(O)NH fragment deviates from planarity because of a twist between the oxime and amide groups. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds, forming zigzag chains in the [013] and [0In the mol\u00adecule of the title compound, C In recent decades, a number of ligands based on 2-hy\u00addroxy\u00adimino\u00adpropane\u00adhydrazide have been obtained. It was shown that such a type of ligand reveals a strong tendency for the formation of polynuclear complexes (Anwar N-[1-(pyrazin-2-yl)ethyl\u00adidene]propano\u00adhydrazide (1), was first described in the work of Feng and co-workers \u00adpropane\u00adhydrazide derivatives. Such a conformation is stabilized additionally by an H4\u22efN5 attractive inter\u00adaction (2.33\u2005\u00c5). Despite the distance being shorter than the sum of the van der Waals radii C(O)NH fragment deviates from planarity (r.m.s. deviation of 0.362\u2005\u00c5) because of a twist between the oxime and the amide groups about the C7\u2014C8 bond. The maximum deviations are 0.8763\u2005(9) and 0.3355\u2005(18)\u2005\u00c5, respectively, for hydrogen (H9C) and non-hydrogen (O1) atoms. The O1\u2014C7\u2014C8\u2014N5 torsion angle is 165.1\u2005(3)\u00b0, significantly less than the average value in 2-(hy\u00addroxy\u00adimino)\u00adpropane\u00adhydrazide derivatives published previ\u00adously [172.1\u2005(4)\u00b0]. Thus, such a twist distortion of the mol\u00adecule seems to be a result of the crystal packing.The CH3.i and C2\u2014H2A\u22efO2ii inter\u00admolecular hydrogen bonds [symmetry codes: (i) \u2212x\u00a0+\u00a0y\u00a0+\u00a01, z\u00a0+\u00a0x\u00a0+\u00a0y\u00a0\u2212\u00a01, z\u00a0\u2212\u00a0A\u22efN2iii inter\u00admolecular hydrogen bonds [symmetry code: (iii) \u2212x\u00a0+\u00a01, \u2212y\u00a0\u2212\u00a01, z\u00a0+\u00a0In the crystal, mol\u00adecules are linked by O2\u2014H2\u22efO1ns Fig.\u00a02. These c4.et al., 2007CrystalExplorer17 from red through white to blue . The Hirshfeld surface of the title compound mapped over dnorm, in the colour range \u22120.6441 to 1.3084 a.u. is shown in Fig.\u00a03A\u22efN2 are the most noticeable inter\u00admolecular inter\u00adactions. In addition, a C2\u2014H2A\u22efO2 weak inter\u00admolecular inter\u00adaction is observed.The Hirshfeld surface analysis pair with the full fingerprint plot outlined in grey. Fig.\u00a04d) reveals two sharp spikes along 1.9\u2005\u00c5 < id + ed < 2.4\u2005\u00c5, which are associated with the O2\u2014H2\u22efO1 hydrogen bond.A fingerprint plot delineated into specific inter\u00adatomic contacts contains information related to specific inter\u00admolecular inter\u00adactions. The blue colour refers to the frequency of occurrence of the level .The calculated geometric parameters are in good agreement with experimental values. It is important to note that the accuracy of the semi-empirical GFN2-xTB method is close to that of the DFT calculations, even though GFN2-xTB calculations are significantly computationally \u2018cheaper\u2019 and C7\u2014N4 is shorter than calculated. Such calculation errors are probably typical for hydrazide derivatives at this level of theory . The maximum number of metal centres per mol\u00adecule for the discrete complexes of this type is 12 \u00adpropane\u00adhydrazide derivatives. Most of them are polynuclear 37.et al., 20181H NMR, 400.13\u2005MHz, (DMSO-d6): 11.97 , 10.21 , 9.31 , 8.56 , 8.55 , 2.37 , 2.02 . IR : 1658 (CO amid I), 1034 (NO oxime). Analysis calculated for C9H11N5O2: C 48.86, H 5.01, N 31.66%; found: C 48.49, H 5.22, N 31.42%.The title compound was prepared according to a slightly modified procedure (Feng 8.Uiso = nUeq of the carrier atom (n = 1.5 for methyl groups and n = 1.2 for other hydrogen atoms).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022007927/vm2270sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022007927/vm2270Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022007927/vm2270Isup3.cdxSupporting information file. DOI: 2195126CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of the title compound was redetermined at 100\u2005K in order to achieve improved structural data, especially with respect to the C\u2014C distances and the hydrogen bonding. 2(C4H9)4Cl2(OH)2], was redetermined at 100\u2005K by use of an area detector to provide new data to improve the structural parameters for detailed analysis. Noteworthy is the folding of the central, non-symmetric, four-membered [SnO]2 ring [dihedral angle about the O\u22efO axis = 1.09\u2005(3)\u00b0], as well as the elongation of the Sn\u2014Cl bonds [mean value = 2.5096\u2005(4)\u2005\u00c5], as a result of inter\u00admolecular O\u2014H\u22efCl hydrogen bonds; the latter lead to a chain-like arrangement of dimeric mol\u00adecules along [101].The structure of the dimeric title compound, [Sn The structure of the title compound has been determined previously at room temperature using point detector data as part of a paper describing the series of dimeric di-tert-butyl\u00adtin(IV) hydroxide halides, [tBu2Sn(OH)X]2 with X = F, Cl and Br I]2 I]2 compounds [pure state: d(Sn\u2014C)mean = 2.190\u2005(3)\u2005\u00c5 mean = 2.193\u2005(10)\u2005\u00c5 to 1.532\u2005(2)\u2005\u00c5 [mean value: 1.527\u2005(4)\u2005\u00c5], Cmeth\u00adyl\u2014C\u2014Cmeth\u00adyl angles in the range 107.1\u2005(1) to 111.1\u2005(1)\u00b0 [mean value: 109.5\u2005(11)\u00b0] and Sn\u2014C\u2014C angles of 107.1\u2005(1) to 111.1\u2005(1)\u00b0 [mean value: 108.9\u2005(12)\u00b0]. These new data are of the same precision and absolute values as those found in the iodide compound both in the pure state , which are considerably longer in comparison with other Br\u00f8nstedt-Base (BB) stabilized diorganotin(IV)-hydroxide-chlorides 2 mol\u00adecules along [101], Fig.\u00a03These unusually long Sn\u2014Cl bonds in the title compound arise from the fact that the chloride atoms are involved in inter\u00admolecular O\u2014H\u22efCl hydrogen bonds Table\u00a01, resultiet al. I. DOI: 10.1107/S2414314623000561/tk4087Isup2.hklStructure factors: contains datablock(s) I. DOI: 2237568CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-indole ring system is nearly planar, while the conformation of the 4H-pyran ring is close to a flattened boat. The mean planes of these fragments are approximately perpendicular to each other. In the crystal, the mol\u00adecules are connected into layers by N\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds.In the title structure, the 2,3-di\u00adhydro-1 17H14BrN3O4, an admixture [0.0324\u2005(11)] of its 7-bromo isomer. The 2,3-di\u00adhydro-1H-indole ring system is nearly planar, while the conformation of the 4H-pyran ring is close to a flattened boat. The mean planes of these fragments form a dihedral angle of 86.67\u2005(9)\u00b0. The carboxyl\u00adate group lies near the plane of 4H-pyran, its orientation is stabilized by an intra\u00admolecular C\u2014H\u22efO contact. In the crystal, the mol\u00adecules are connected into layers by N\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds. The most important contributions to the crystal packing are from H\u22efH (33.1%), O\u22efH/H\u22efO (16.3%), N\u22efH/H\u22efN (12.1%), Br\u22efH/H\u22efBr (11.5%) and C\u22efH/H\u22efC (10.6%) inter\u00adactions.The crystal used for structure determination contained, along with the title compound, C The mean planes of the 2,3-di\u00adhydro-1H-indole system and the 4H-pyran ring are approximately perpendicular to each other, forming a dihedral angle of 86.67\u2005(9)\u00b0. The carboxyl\u00adate group lies near the plane of 4H-pyran, with O3\u2014C13\u2014C10\u2014C11 and O4\u2014C13\u2014C10\u2014C3 torsion angles of \u221213.4\u2005(3) and \u22128.8\u2005(2)\u00b0, respectively. An intra\u00admolecular C16\u2014H16A\u22efO3 contact stabilizes the conformation of the mol\u00adecule ring motif :0.0324\u2005(11) ratio, whereas the positions of other atoms of these isomers coincide with each other Fig.\u00a02. The 2,3le Fig.\u00a02, generat3.H-pyran and benzene rings link adjacent mol\u00adecules within these layers Table\u00a01 \u25b8 \u25b8. ICrystalExplorer17 ring motif. The mol\u00adecules are linked by pairs of N\u2014H\u22efO hydrogen bonds into ribbons extending along the b-axis direction and consisting of In the crystal of II, the six-membered pyran ring adopts a conformation close to a flattened boat, as in the title structure. The mol\u00adecules are joined by pairs of N\u2014H\u22efN hydrogen bonds into dimers, those are linked by N\u2014H\u22efO contacts to form ribbons along the a-axis direction.In the crystal of III, the pyran ring is nearly planar. The mol\u00adecules are joined by pairs of N\u2014H\u22efN hydrogen bonds into centrosymmetric dimers, which are linked by N\u2014H\u22efO contacts into ribbons along the c-axis direction.In the crystal of 5.The title compound was synthesized using the reported procedure , N8\u2014H8A = 0.88\u2005(3) and N8\u2014H8B = 0.86\u2005(3)\u2005\u00c5], but their isotropic displacement parameters were constrained to take a value of 1.2Ueq(N). All H atoms bound to C atoms were positioned geometrically and refined as riding with C\u2014H = 0.95 (aromatic), 0.99 (methyl\u00adene) and 0.98\u2005\u00c5 (meth\u00adyl), withUiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for all others.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989022008271/yk2174sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022008271/yk2174Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022008271/yk2174Isup3.cmlSupporting information file. DOI: 2202347CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The compound crystallizes with one mol\u00adecule in the asymmetric unit in the monoclinic space group P21/n with a rather long b axis [33.8752\u2005(2)\u2005\u00c5]. Weak C\u2014H\u22efO and C\u2014H\u22efN inter\u00adactions consolidate the crystal packing. The nitrite-O atoms each occupy a single position in the coordination geometry.The mol\u00adecular structure of the title compound, [Ag(NO The nitrito ligand coordinates in a near symmetric fashion with similar bond lengths . The pseudo-tetra\u00adhedral coordination environment exhibited around the AgI atom stems from the three coordinating ligands, with corresponding bond angles of P1\u2014Ag1\u2014P2 [124.597\u2005(16)\u00b0], P1\u2014Ag1\u2014O1 [116.26\u2005(6)\u00b0], P1\u2014Ag1\u2014O2 [125.62\u2005(4)\u00b0], P2\u2014Ag1\u2014O1 [107.68\u2005(7)\u00b0], and P2\u2014Ag1\u2014O2 [107.83\u2005(4)\u00b0]. The bidentate coordination of the nitrito ligand is underpinned by the O1\u2014Ag1\u2014O2 bite angle of 50.80\u2005(7)\u00b0. The ipso-aryl carbon atoms of each of the phosphine ligands overlap in a near-staggered fashion when viewed down the P1\u2014Ag1\u2014P2 axis, presumably due to the steric effect of the bulky phosphine ligands. Corresponding torsion angles are P2\u2014Ag1\u2014P1\u2014C1 = 9.90\u2005(7)\u00b0, P2\u2014Ag1\u2014P1\u2014C8 = \u2212108.02\u2005(8)\u00b0, P2\u2014Ag1\u2014P1\u2014C15 = 128.73\u2005(9)\u00b0, P1\u2014Ag1\u2014P2\u2014C22 = \u2212172.57\u2005(7)\u00b0, P1\u2014Ag1\u2014P2\u2014C36 = 70.75\u2005(8)\u00b0, and P1\u2014Ag1\u2014P2\u2014C29 = \u221247.35\u2005(7)\u00b0. All of the aforementioned bond lengths and angles closely correspond to those of related AgI phosphine complexes and silver nitrite (1\u2005mmol) were dissolved separately in aceto\u00adnitrile (10\u2005ml). The two solutions were carefully mixed together and heated to 353\u2005K for approximately 2\u2005h. The solution was left to crystallize, and small clear colourless crystals were obtained.Tris-Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314622011488/wm4174sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622011488/wm4174Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622011488/wm4174Isup3.cdxSupporting information file. DOI: 2223249CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure, the cations and anions are inter\u00adconnected via several N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds.The title compound, C 6H2N3O7\u2212\u00b7C10H13Cl2N2+, crystallizes with one 1-piperazine (DP) cation and one picrate (PA) anion in the asymmetric unit. In the crystal structure, the DP cation and PA anion are inter\u00adconnected via several N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. The DP cation and PA anion are further connected through C\u2014Cl\u22ef\u03c0 and N\u2014O\u22ef\u03c0 [3.7814\u2005(4)\u2005\u00c5] inter\u00adactions. The DP cations are further inter\u00adconnected via a weak inter\u00admolecular Cl\u22efCl [3.2613\u2005(4)\u2005\u00c5] halogen\u2013halogen inter\u00adaction. The combination of these supra\u00admolecular inter\u00adactions leads to a herringbone like supra\u00admolecular architecture.The title compound, C Inversion-related cation\u2013anion pairs are also linked through N1\u2014H1A\u22efO4, N1\u2014H1B\u22efO2 and C17\u2014H17\u22efO1 hydrogen bonds, forming an Cg1, C\u2014Cl\u22efCg3v and N\u2014O\u22efCg3; symmetry codes: (v) 1\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z] with C\u22ef\u03c0 distances of 3.8201\u2005(4) and 3.7785\u2005(4)\u2005\u00c5, and N\u22ef\u03c0 = 3.782\u2005(2)\u2005\u00c5, with C\u2014Cl\u22ef\u03c0 angles of 74.15\u2005(7) and 76.91\u2005(7)\u00b0 and an N\u2014O\u22ef\u03c0 angle of 68.80\u2005(12)\u00b0. The combination of N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014Cl\u22ef\u03c0 and N\u2014O\u22ef\u03c0 inter\u00adactions leads to the formation of a three-dimensional supra\u00admolecular herringbone architecture, which propagates along the a- and c-axis directions inter\u00adactions [3.2613\u2005(4)\u2005\u00c5] piperazine (DP) and picric acid (PA) were dissolved independently in water and ethanol. The reactants were then mixed together in a 100\u2005ml beaker and heated over a water bath at 90\u00b0C for 1\u2005h Fig.\u00a05. The cleCrystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621003795/bv4037sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314621003795/bv4037Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314621003795/bv4037Isup3.cmlSupporting information file. DOI: 2076126CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the pento\u00adfuran\u00adose ring has a twisted conformation while the other five-membered ring has an envelope conformation; the two hy\u00addroxy groups C are involved in an infinite network of O\u2014H\u22ef O bonds, forming a layer parallel to the (001) plane. 8H14O5, the pento\u00adfuran\u00adose five-membered ring has a twisted conformation on two carbon atoms while the five-membered ring of the iso\u00adpropyl\u00adidene group has an envelope conformation on an oxygen atom. Hy\u00addroxy groups are involved an infinite network of O\u2014H\u22efO hydrogen bonds that leads to the formation of a layer parallel to the (001) plane. Only weak C\u2014H\u22efO contacts exist between neighboring layers.In the title compound C Our inter\u00adest in 1 stems from the possibility of conducting de\u00adoxy\u00adgenation at its C3 position to obtain 3-de\u00adoxy-1,2-O-iso\u00adpropyl\u00adidene-\u03b2-d-threo-pento\u00adfuran\u00adose as a chiral synthon for further synthetic work side to furnish 1,2-O-iso\u00adpropyl\u00adidene-5-O\u2013t-butyl\u00addiphenyl\u00adsilyl-\u03b2-d-lyxo-furan\u00adose, whose desilylation gave the target 1. It should be pointed out that under these iso\u00adpropyl\u00adidenation conditions, d-lyxose furnished only its \u03b1,\u03b2-2,3-O-iso\u00adpropyl\u00adidene\u00adfuran\u00adose \u2005\u00c5, \u03c6 = 117.6\u2005(2)\u00b0]. The five-membered ring of the iso\u00adpropyl\u00adidene group has an envelope conformation on atom O1 . Puckering parameters distance is 3.389\u2005(2)\u2005\u00c5. Similar hydrogen bonds have been observed in various carbohydrates plane Table\u00a01. Only wep Table\u00a01 may addiet al. and a probability of the absolute configuration being correct of 1.000.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314620016302/zq4044sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314620016302/zq4044Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314620016302/zq4044Isup3.cdxSupporting information file. DOI: 10.1107/S2414314620016302/zq4044sup4.txtCuK CIF file for chirality confirmation. DOI: 2050681CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds connect the mol\u00adecules into [001] chains. 12H9ClN2O, the dihedral angle between the aromatic rings is 1.78\u2005(4)\u00b0 and an intra\u00admolecular O\u2014H\u22efN hydrogen bond closes an S(6) ring. In the crystal, C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds connect the mol\u00adecules into [001] chains.In the title compound, C S(6) ring. The disposition of the aromatic rings is trans as indicated by the C2\u2014N2\u2014C13\u2014C7 torsion angle of \u2212179.7\u2005(2)\u00b0 \u00b0 and an intra\u00admolecular O\u2014H\u22efN hydrogen bond closes an )\u00b0 Fig.\u00a01. In the )\u00b0 Fig.\u00a01 generateps Fig.\u00a02. These aps Fig.\u00a02.et al. and 2-hy\u00addroxy benzaldehyde (1.2\u2005mmol) were mixed in 20\u2005ml of absolute ethanol with the addition of few drops of piperidine as catalyst. The mixture was refluxed for 5\u2005h at 60\u201370\u00b0C. Colourless blocks of the title compound were obtained from the mother solution on cooling.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314620000115/hb4333sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314620000115/hb4333Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314620000115/hb4333Isup3.cmlSupporting information file. DOI: 1975774CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-021-00096-x, published online 19 October 2021Correction to: The original version of this Article contained an error in the legend of Figure\u00a01.A) Effect on IFN-\u03b3 (a) and IL-10 expression (b) and IFN-\u03b3 to IL-10 expression ratio (c) in LPS-stimulated macrophage cells. (B) Effect on IFN-\u03b3 (a) and IL-10 expression (b) and IFN-\u03b3 to IL-10 expression ratio (c) in resident macrophage cells. Macrophage cells (1\u2009\u00d7\u2009106/mL) isolated from peritoneal cavity were incubated with\u00a0Lactobacillus rhamnosus\u00a0NK210 or\u00a0Bifidobacterium longum\u00a0NK219 in the absence or presence of LPS. Normal control group (CON) was treated with saline instead of LPS. Data values were described as mean\u2009\u00b1\u2009SD (n\u2009=\u20094). Data values indicate mean\u2009\u00b1\u2009SD.\u00a0#p\u2009<\u20090.05 versus NC group. *p\u2009<\u20090.05 versus LP group.\u201d\u201cEffect of NK210 and NK219 on the expression of interferon (IFN)-\u03b3 and IL-10 in macrophages stimulated with or without LPS. (now reads:A) Effect on IFN-\u03b3 (a) and IL-10 expression (b) and IFN-\u03b3 to IL-10 expression ratio (c) in resident macrophage cells. (B) Effect on IFN-\u03b3 (a) and IL-10 expression (b) and IFN-\u03b3 to IL-10 expression ratio (c) in LPS-stimulated macrophage cells. Macrophage cells (1\u2009\u00d7\u2009106/mL) isolated from peritoneal cavity were incubated with\u00a0Lactobacillus rhamnosus\u00a0NK210 or\u00a0Bifidobacterium longum\u00a0NK219 in the absence or presence of LPS. Normal control group (CON) was treated with saline instead of LPS. Data values were described as mean\u2009\u00b1\u2009SD (n\u2009=\u20094). Data values indicate mean\u2009\u00b1\u2009SD.\u00a0#p\u2009<\u20090.05 versus NC group. *p\u2009<\u20090.05 versus LP group.\u201d\u201cEffect of NK210 and NK219 on the expression of interferon (IFN)-\u03b3 and IL-10 in macrophages stimulated with or without LPS. (The original Article has been corrected."} +{"text": "In the title Schiff base, the dihedral angle between the phenyl rings of the benzil unit is 74.14\u2005(5)\u00b0. 20H14ClNO, obtained from the reaction of 4-chloro aniline with benzil, has an approximate T shape. The dihedral angle between the phenyl rings of the benzil unit is 74.14\u2005(15)\u00b0. The extended structure features C\u2014H\u22efO hydrogen bonds.The title Schiff base, C The C1\u2013C6 phenyl ring makes dihedral angles of 20.56\u2005(6) and 74.03\u2005(6)\u00b0with the C9\u2013C10 and C15\u2013C16 phenyl ring, respectively, of the benzil unit. The dihedral angle between the phenyl rings of the benzil unit is 74.14\u2005(5)\u00b0. The C\u2014N iminium bond length [1.268\u2005(3)\u2005\u00c5] is comparable to that observed in (E)-1-[4-({4-[(4-meth\u00adoxy\u00adbenzyl\u00adidene)amino]\u00adphen\u00adyl}sulfan\u00adyl)phen\u00adyl]ethan-1-one . Together, these hydrogen bonds lead to the formation of a three-dimensional network. Aromatic \u03c0\u2013\u03c0 stacking generates inversion dimers featuring the C15\u2013C20 phenyl rings with a centroid\u2013centroid distance of 3.744\u2005(3)\u2005\u00c5 [Fig.\u00a03b)]. Along the c-axis direction, weak C\u2014H\u22ef\u03c0(ring) inter\u00adactions occur.In the crystal, the mol\u00adecules are aligned head-to-foot along the g Table\u00a01. The C18Crystal Explorer 3.1 and shape-index. The red spots in Fig.\u00a04a) reflect the formation of C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions. In the shape-index map [Fig.\u00a04b)], the adjacent red and blue triangle-like patches represent concave regions that indicate C\u2014H\u22ef\u03c0(ring) and \u03c0\u2013\u03c0 stacking inter\u00adactions. The two-dimensional FP plots indicate that the most important contributions to the packing, in descending percentage contribution, are from H\u22efC (37.7%), H\u22efH (34.6%), H\u22efCl (14.0%), H\u22efO (6.1%), H\u22efN (4.0%) and C\u22efC (1.9%) contacts.A Hirshfeld surface (HS) analysis was performed and the associated two-dimensional fingerprint (FP) plots , 1660 (C=O), 3064 (aromatic C\u2014H), 1212 (C\u2014N) and 718 (C\u2014Cl).To a solution of benzil and 1\u2005ml of acetic acid in ethanol (20\u2005ml) was added 4-chloro aniline (0.01\u2005mmol) dissolved in ethanol (15\u2005ml). The mixture was stirred for 3\u2005h under reflux. The product was isolated, recrystallized from ethanol solution and then dried in a vacuum to give the title compound . Yellow single crystals suitable for X-ray analysis were obtained by slow evaporation of a ethanol solution. IR \u03bd, cmCrystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314623000652/hb4414sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314623000652/hb4414Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623000652/hb4414Isup3.cmlSupporting information file. DOI: 2237868CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Compounds 1\u20133 are obtained by reacting Me\u2010cAAC=PK (amino)carbene) and dihaloaryl borane in toluene. All three compounds were characterized by X\u2010ray crystallography. Quantum mechanical studies indicated that these compounds have two lone pairs on the P center viz., an \u03c3\u2010type lone pair and a \u201chidden\u201d \u03c0\u2010type lone pair. Hence, these compounds can act as double Lewis bases, and the basicity of the \u03c0\u2010type lone pair is higher than the \u03c3\u2010type lone pair.Herein, the synthesis and characterization of the carbene\u2010stabilized boryl phosphinidenes What hides within: Three different singlet boryl phosphinidenes were synthesized by using cAAC carbene, and their inversely polarized phosphaalkene characteristics were investigated. Quantum mechanical studies suggest that the P center of these boryl phosphinidenes contains two lone pairs viz., an \u03c3\u2010type lone pair and a \u201chidden\u201d \u03c0\u2010type lone pair. FoiPr2NP\u0308:), which was formed as a transient intermediate while thermolysis of dibenzo\u20107\u2010phosphanorbornadiene (Scheme\u2005h).i). The reactivities of phosphino\u2010phosphinidene were studied using several nucleophiles such as CO, isocyanides, carbenes, and phosphines, etc. \u20131\u20133) which are also characterized as inversely polarized phosphaalkenes. DFT calculations are employed to obtain further insight into the bonding and structure\u2013property relationship of the newly developed carbene\u2010stabilized boryl phosphinidenes.In 2012, Cummins et\u2005al. reported a mono\u2010coordinated phosphinidene in a 1\u2009:\u20091 molecular ratio.Compounds 1, 2, and 3 were obtained in good yield at 0\u2009\u00b0C from the concentrated toluene solution. Compounds 1\u20133 were fully characterized by NMR and mass spectrometry (LIFDI). All these compounds are highly stable in the solid\u2010state and in solutions at room temperature under an inert atmosphere for months. However, they are unstable under ambient conditions and slowly change their intense yellow color to a colorless form. Single\u2010crystal X\u2010ray diffraction and elemental analysis confirmed the compositions of 1, 2, and 3.The block\u2010shaped yellow crystals of 13C, 31P, and 11B NMR spectroscopy provided a wealth of structural information on the inversely polarized phosphaalkene. The 13C{1H} signal of C=P unit of inversely polarized phosphaalkenes appeared at 220.9 , 220.8 , and 220.7 ppm for compounds 1, 2, and 3 respectively. The observed 13C{1H} NMR signals and JC\u2010P coupling constants for the C=P units are similar to the reported phosphaalkenes.31P NMR shifts for the compounds are strongly affected by the electronic nature of the substituents and the hybridization of the phosphorus atom. The introduction of an electropositive boryl group provides a deshielding effect on the phosphorus chemical shift. Also, the halogen atoms such as Cl, Br, and I on the boron center typically move the chemical shift significantly downfield. The 31P{1H} NMR signal appeared at 35.5, 47.1, and 66.9\u2005ppm for compounds 1, 2, and 3, respectively. This observation is in accordance with the increasing order of Lewis acidity of the boron center, which increases when the halogen changes from Cl to I.31P{1H} NMR spectroscopy supports the formation of delocalized C\u2212P\u2212B \u03c0\u2010system and the extent of delocalization increases from compound 1 to 3. Note that the 31P chemical shift values move downfield compared to inversely polarized phosphaalkenes.31P chemical shifts for phosphatriafulvenes show inverse polarity around \u221223.2 to \u221274.1\u2005ppm.11B{1H} nucleus and the asymmetric local fields in compounds usually result in broad 11B{1H} NMR resonances. The 11B{1H} NMR resonances appear at 72.83, 71.81, and 69.80\u2005ppm for the compounds 1, 2, and 3, respectively. The 31P and 11B NMR data indicate that the B\u2212P bond has considerable double\u2010bond character and 31P and 11B resonances are similar to those reported phosphinidene boranes (R\u2212B=P\u2212R).1, 2, and 3 consist of a C\u2212P\u2212B backbone with an almost coplanar halogen atom \u00b0 (1), 179.02 (6)\u00b0 (2), and 177.70 (6)\u00b0 (3). The C=P=B skeleton is significantly bent with a bond angle of 114.68\u00b0, 111.94\u00b0, 110.78\u00b0 and the dihedral angles between C17\u2212C16\u2212N1 and X1\u2212B1\u2212C1 planes are 25.06\u00b0, 21.43\u00b0 and 31.50\u00b0 for compounds 1, 2, and 3, respectively.The solid\u2010state structures of complexes cis to the C\u2212P bond (C16\u2212P1 1.7709 (15)\u2005\u00c5) in complex 1, whereas the B\u2212X bonds in complexes 2 (B1\u2212Br1 2.0085(15)\u2005\u00c5), and 3 (B1\u2212I1 2.2504 (15)\u2005\u00c5) are in trans position to the C\u2212P bonds, respectively. Furthermore, the nearly anti\u2010periplanar alignment represented by torsion angles N\u2212C\u2212P\u2212B (1 172.74(12)\u00b0, 2 164.82(9)\u00b0, 3 157.78(10)\u00b0) decrease from species 1 to 3. The P1\u2212C16 (1 1.7709 (15)\u2005\u00c5, 2 1.7850 (14)\u2005\u00c5, 3 1.7944(14)\u2005\u00c5) bond lengths are longer than reported double bond lengths in phosphaalkenes (1.61\u20131.71\u2005\u00c5)The molecular structures reveal that in the solid state, the B1\u2212Cl1 (1.8046(18)\u2005\u00c5) bond is 1 1.8607(18)\u2005\u00c5, 2 1.8607(15)\u2005\u00c5, 3 1.8456(15)\u2005\u00c5) bond lengths are elongated compared to phosphaborenes (1.8211(16)\u20131.8309(16)\u2005\u00c5)tert\u2010butyl\u20101\u2010phosphaethene)(diethylhydroborato)titanocene] (2.054(3)\u2005\u00c5)3 complex (1.931(4)\u20131.943(4)\u2005\u00c5)1\u20133), the most elongated P\u2212C bond length (1.7944(14)\u2005\u00c5) and the shortest B\u2212P bond length (1.8456(15)\u2005\u00c5) were found in 3 (X=I). This is due to the high Lewis acidity of the boron center of compound 3 compared to 1\u20132 (X=Cl and Br), and the phosphinidene lone pair being relatively more engaged in donating to the boron center and compared to back\u2010bonding to the carbene center of cAAC. Additionally, Ccarbene\u2212N bond lengths in 1\u20133 (1 1.3352(19)\u2005\u00c5, 2 1.3379(16)\u2005\u00c5, 3 1.3306(16)\u2005\u00c5) are elongated (1.9\u20132.5\u2009%) compared to that in free cAAC (1.3053(13)\u2005\u00c5),Similarly, P1\u2212B1 to I . The isomer 1\u2032, where the B\u2212Cl bond is oriented similar to B\u2212X bonds in complexes 2 and 3, is less stable by 1.82\u2005kcal\u2009mol\u22121. It can be attributed to the shorter P\u2212Cl distance (2.97\u2005\u00c5), resulting in strong repulsions between the lone pairs on P and Cl centers .Quantum mechanical calculations were carried out at the M06\u20102X/def2\u2010TZVPP//BP86/def2\u2010TZVP level of theory by incorporating Grimme empirical dispersion correction with Becke\u2013Johnson damping (GD3BJ) to explore the bonding and reactivity of the molecules 1 reveals that HOMO\u20102 is a \u03c3\u2010type lone pair orbital with a larger contribution from the P atom , P\u2212B (1.31\u20131.41) and Ccarbene\u2212N (1.29\u20131.32) bonds. Note that these Wiberg bond indices are lower than the Ccarbene\u2212N Wiberg bond index of free cAAC . The difference in electronegativity also induces polarity in the Ccarbene\u2212P, P\u2212B and Ccarbene\u2212N bonds, reducing the bond index values. These bond index values support the partial double bond character observed in these bonds. The overall high positive group charge on cAAC (0.14\u20130.18) implies that \u03c3\u2010donation from cAAC is higher than the \u03c0\u2010back donation from the P center. The NBO analysis shows considerable polarization of the P\u2212CcAAC \u03c3\u2010MO and \u03c0\u2010MO in opposite directions (Table\u2005S8), thus indicating the inversely polarized phosphaalkene nature. However, the net polarization is minimal, as seen from the natural charges on CcAAC (0.10\u20130.13 e) and P (0.06\u20130.08 e).The molecular orbital (MO) analysis of \u22121 when halogen atom is Cl (1), 2.6\u2005kcal\u2009mol\u22121 for Br (2), and 2.7\u2005kcal\u2009mol\u22121 for I (3). On the other hand, the energetics corresponding to the donation from halogen to B center is significantly high and decreases as 40.7\u2005kcal\u2009mol\u22121 for 1, 35.3\u2005kcal\u2009mol\u22121 for 2, and 30.31\u2005kcal\u2009mol\u22121 for 3. The strength of the donation from the halogen atom is a deciding factor for the P\u2212B and B\u2212X bond lengths. The P\u2212B bond length is shortest for 3 (1.860\u2005\u00c5); this is in agreement with the weaker donation of I lone pairs to the formally empty p\u2010orbital on B. Note that the strength of the donation from P\u2212C \u03c0\u2010NBO to the B center is also significant viz., 38.4\u2005kcal\u2009mol\u22121 for 1, 49.6\u2005kcal\u2009mol\u22121 for 2, and 56.0\u2005kcal\u2009mol\u22121 for 3. The balance between these second\u2010order interactions results in the intermediate dihedral angles between the planes defined by terminal B and C atoms with their corresponding substituents (23.5\u201333.7\u00b0).Three major interactions are responsible for the geometry of the compounds viz., the donation of \u03c3\u2010lone pair on P to B center, the donation of lone pair of electrons from halogen atom to B center, and the donation from P\u2212C \u03c0\u2010NBO to the B center. The strength of interaction corresponding to the donation of \u03c3\u2010lone pair on P to B center is 5.4\u2005kcal\u2009mol1\u20133 is monovalent, forming an electron sharing bond with B center while accepting a lone pair of electrons from cAAC. The bonding is schematically represented in Figure\u2005\u22121 . Hence, the P atom can act as a coordinating center for accepting electrons from cAAC. The calculated dissociation energy of compounds 1\u20133 to respective singlet boryl phosphinidene and cAAC was found to be moderately high, indicative of the strong binding between P\u2212C centers . P\u2212C bond dissociation energy values for carbene stabilized phosphinidenes have been calculated to fall in the range 43\u201375\u2005kcal\u2009mol\u22121 at BP86/TZ2P level of theory.Based on the geometrical, MO and NBO analyses, the P center in \u22121) and close to that of NHCs and C(PH3)2 but less than that in carbodicarbenes, C(NHCMe)2 .1\u20133. This suggests that the delocalized \u03c0\u2010MO in 1\u20133 is more basic than the \u03c3\u2010lone pair having a very high s\u2010character (53.8\u201354.4\u2009%). Hence, the delocalized \u03c0\u2010MO can be considered as a hidden lone pair. Frenking and co\u2010workers have used the term hidden lone pair for a delocalized \u03c0\u2010MO in carbodicarbenes and related C(0) compounds, which show very high proton affinity.A bent geometry with an active \u03c3\u2010lone pair and a delocalized \u03c0\u2010MO is the prominent feature of carbones and related species.carbene\u2212P (0.7\u20131.3\u2009%) and P\u2212B (2.5\u20132.7\u2009%) bonds and a shortening of the Ccarbene\u2212N bond . This corroborated well with increased cAAC group charge (0.49\u20130.52) and decreased negative natural charge on N (\u22120.35) atom. The MO analysis indicates an active lone pair (35.0\u201338.2\u2009% s character) with a major coefficient on the P atom . The presence of an active lone pair with relatively low s\u2010character induces the addition of the second proton at the P center. The calculated second proton affinity is also quite high , indicating that P (I) center in molecules 1\u20133 can act as a double Lewis base.The first protonation induces an elongation in C1\u20133 feature a P(I) center with two lone pairs, viz. a highly reactive \u201chidden\u201d \u03c0\u2010lone pair and a \u03c3\u2010lone pair.In conclusion, we have designed, synthesized, and characterized three cAAC\u2010stabilized boryl\u2010phosphinidenes, which also show inversely polarized phosphaalkene characteristics. Quantum mechanical calculations reveal that compounds Essential experimental: Crystal data for 1 at 100(2) K: C35H54Cl2BClNP, Mr=566.02\u2005g/mol, 0.401\u00d70.303\u00d70.139\u2005mm, monoclinic, P21/n, a=9.810(2)\u2005\u00c5, b=17.822(3)\u2005\u00c5, c=19.840(3)\u2005\u00c5, \u03b2=92.64(2)\u00b0, V=3465.0(11)\u2005\u00c53, Z=4, \u03bc(MoK\u03b1)=0.179\u2005mm\u22121, \u03b8max=25.510\u00b0, 78252 reflections measured, 6411 independent (Rint=0.0530), R1=0.0364 [I>2\u03c3(I)], wR2=0.0943 , res. density peaks: 0.383 to \u22120.237\u2005e\u2009A\u22123, CCDC: 2058117.2 at 100(2) K: C35H54BBrNP, Mr=610.48\u2005g/mol, 0.584\u00d70.317\u00d70.309\u2005mm, triclinic, P1\u203e, a=9.805(2)\u2005\u00c5, b=11.181(2)\u2005\u00c5, c=16.560(3)\u2005\u00c5, \u03b1=73.82(2)\u00b0, \u03b2=82.35(2)\u00b0, \u03b3=88.07(3)\u00b0, V=1728.0(6)\u2005\u00c53, Z=2, \u03bc(MoK\u03b1)=1.258\u2005mm\u22121, \u03b8max=27.934\u00b0, 53463 reflections measured, 8274 independent (Rint=0.0469), R1=0.0265 [I>2\u03c3(I)], wR2=0.0674 , res. density peaks: 0.368 to \u22120.357\u2005e\u2009A\u22123.Crystal data for 3 at 100(2) K: C42H62BINP, Mr=749.60\u2005g/mol, 0.341\u00d70.253\u00d70.235\u2005mm, triclinic, P1\u203e, a=10.676(2)\u2005\u00c5, b=12.167(2)\u2005\u00c5, c=16.836(3)\u2005\u00c5, \u03b1=107.24(2)\u00b0, \u03b2=94.60(2)\u00b0, \u03b3=101.62(3)\u00b0, V=2022.8(7)\u2005\u00c53, Z=2, \u03bc(Mo K\u03b1)=0.859\u2005mm\u22121, \u03b8max=27.200\u00b0, 82576 reflections measured, 9022 independent (Rint=0.0303), R1=0.0213 [I>2\u03c3(I)], wR2=0.0531 , res. density peaks: 0.452 to \u22120.462\u2005e\u2009A\u22123.Crystal data for \u03bb correction1 was applied using SADABS.F2 using SHELXLAll crystals were selected under cooling by using an X\u2010Temp2 device.1), 2058118 (2), and 2058119 (3)2058117 should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "In the title 1:2 co-crystal, C\u2014I\u22efN halogen bonds between one of the 1,2,3,5-tetra\u00adfluoro-4,6-di\u00adiodo\u00adbenzene mol\u00adecules and the 5-{[4-(di\u00admethyl\u00adamino)\u00adphen\u00adyl]ethyn\u00adyl}pyrimidine mol\u00adecule form [110] chains while the second 1,2,3,5-tetra\u00adfluoro-4,6-di\u00adiodo\u00adbenzene mol\u00adecule resides in [100] channels. 14H13N3\u00b72CF4I2. In the extended structure, two unique C\u2014I\u22efN halogen bonds from one of the 1,2,3,5-tetra\u00adfluoro-4,6-di\u00adiodo\u00adbenzene mol\u00adecules to the pyrimidine N atoms of the 5-{[4-(di\u00admethyl\u00adamino)\u00adphen\u00adyl]ethyn\u00adyl}pyrimidine mol\u00adecule generate [110] chains and layers of these chains are \u03c0-stacked along the a-axis direction. The second 1,2,3,5-tetra\u00adfluoro-4,6-di\u00adiodo\u00adbenzene mol\u00adecule resides in channels formed parallel to the a-axis direction between stacks of 5-{[4-(di\u00admethyl\u00adamino)\u00adphen\u00adyl]ethyn\u00adyl}pyrimidine mol\u00adecules and inter\u00adacts with them via C\u2014I\u22ef\u03c0 contacts.The treatment of 5-{[4-(di\u00admethyl\u00adamino)\u00adphen\u00adyl]ethyn\u00adyl}pyrimidine with a threefold excess of 1,2,3,5-tetra\u00adfluoro-4,6-di\u00adiodo\u00adbenzene in di\u00adchloro\u00admethane solution led to the formation of the unexpected 1:2 title co-crystal, C The APEP and the halogen-bonded 13DIFP mol\u00adecule are essentially coplanar: the inter\u00adplanar angle between the pyrimidine ring and the amino\u00adphenyl ring is 4.24\u2005(15)\u00b0 and the inter\u00adplanar angle between the pyrimidine ring and the halogen-bonded 13DIFP mol\u00adecule is 6.63\u2005(15)\u00b0. The two unique C\u2014I\u22efN halogen bonds that combine to form a zigzag alternating halogen-bonded chain, shown in Fig.\u00a02i = 2.853\u2005(2) and 2.901\u2005(2)\u2005\u00c5 and angles C15\u2014I1\u22efN1 and C17\u2014I2\u22efN2i = 174.8\u2005(9) and 173.8\u2005(8)\u00b0, respectively . These distances and angles are similar to those previously reported in the 1:1 co-crystal formed between these two mol\u00adecules of 2.920\u2005(2)\u2005\u00c5 and 178.27\u2005(6)\u00b0 1\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z] separations of 3.276\u2005(3) and 3.316\u2005(3)\u2005\u00c5, respectively. These are significantly less than the sum of the van der Waals radii of 3.68\u2005\u00c5 at 89 and 90%, respectively. The second I atom has close I\u22efF contacts to two neighboring 13DIFB mol\u00adecules with I3\u22efF6iii and I3\u22efF3iv separations of 3.2142\u2005(17) and 3.30129\u2005(15)\u2005\u00c5 as compared to the sum of the van der Waals radii of 3.38\u2005\u00c5 .The pair of loosely \u03c0-stacked 13DIFB mol\u00adecules inter\u00adact with the surrounding mol\u00adecules as shown in the Hirshfeld surface plot in Fig.\u00a05The pyrimidine APEP (8.3\u2005mg) was dissolved in 2\u2005ml of di\u00adchloro\u00admethane in a screw-cap vial. Three equivalents of 13DIFB were added and the solvent was allowed to slowly evaporate until crystals formed when the vial was sealed to prevent further loss of solvent.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2414314622003807/hb4404sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622003807/hb4404Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622003807/hb4404Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2414314622003807/hb4404Isup4.cmlSupporting information file. DOI: 2164881CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Two of the copper(I) atoms are additionally linked to PMe3 ligands, giving a distorted trigonal coordination.The mol\u00adecular structure of the title complex consists of an eight-membered Cu 4(C9H11)4(C3H9P)2] or [Cu4(Mes)4(PMe3)2] , was synthesized from copper(I) mesityl and tri\u00admethyl\u00adphosphane in THF as solvent. The mol\u00adecular structure of the complex has C2 symmetry and consists of four copper(I) atoms bridged by four \u03bc-mesityl groups, giving an eight-membered puckered {Cu4C4} ring. Additionally, two copper(I) atoms at opposite corners of the Cu4 rhomb are each linked to a terminal PMe3 ligand. The PMe3-bearing copper(I) atoms exhibit a distorted trigonal\u2013planar coordination mode whereas the remaining Cu atoms linked to two mesityl groups are nearly linearly coordinated.The title compound, [Cu I organyls, mesitylcopper is one of the most extensively studied compounds. Since its first synthesis in 1981 , the reaction proceeds under retention of the tetra\u00adnuclear cluster structure to form [Cu4(Mes)4(THT)2] 2]\u00b7C7H8 with a yet unknown crystal structure 2Cu][CuMes2] 4(PMe3)2] (1).In order to get some insight into the reactivity of mesitylcopper towards sterically less demanding phosphanes, tri\u00admethyl\u00adphosphane was chosen as a ligand. Treatment of a solution of mesitylcopper in THF with PMe1) comprises four copper(I) atoms that are linked by four \u03bc-mesityl groups to give an eight-membered {Cu4C4} ring \u20132.4625\u2005(5)\u2005\u00c5] suggest cuprophilic inter\u00adactions. The Cu\u22efCu separations between the copper atoms at opposite corners of the rhomb are 4.2013\u2005(5)\u2005\u00c5 for Cu2\u22efCu2i , [Cu4(o-Tol)4(SMe2)2] 2] exhibits two types of differently coordin\u00adated Cu atoms \u00b0] is clearly more pronounced than in [Cu4Mes4] (164.05\u2013165.70\u00b0). Apart from two mesityl groups, Cu2 bears a PMe3 unit as a third ligand. The increased coordination number leads to a further enlargement of the Cu\u2014C distances with values of 2.093\u2005(3) and 2.095\u2005(3)\u2005\u00c5. The coordination around Cu2 is planar with a C\u2014Cu\u2014C angle of 163.0\u2005(1)\u00b0 and C\u2014Cu\u2014P angles of 97.9\u2005(1)\u00b0 and 99.0\u2005(1)\u00b0 (sum of the angles around Cu2: 359.9\u00b0). Comparison of the bond lengths of compound (1) and related [Cu4Mes4L2] complexes reveals that the ligand PMe3 leads to a larger increase of the Cu\u2014C distances for the tricoordinate copper atoms than other ligands investigated so far. In [Cu4Mes4L2] complexes with L = piperidine, allyl methyl sulfide, 2,5-di\u00adthia\u00adhexane, tetra\u00adhydro\u00adthio\u00adphene and bis\u00ad{2-[1-(di\u00admethyl\u00adamino)\u00adeth\u00adyl]phenyl\u00adthiol\u00adato}magnesium, the mean Cu\u2014C distances for the tricoordinated copper atoms are in the range 2.054\u20132.064\u2005\u00c5. In the case of the dicoordinated Cu there is no particular effect. Furthermore, there is a slight influence on the C\u2014Cu\u2014C angles for the dicoordinated [138.3\u2005(1)\u00b0] and the tricoordinate copper atoms [163.0\u2005(1)\u00b0], which are smaller than in the [Cu4Mes4L2] complexes mentioned above .The mol\u00adecular structure of = 2.87\u2005\u00c5, C17\u2014H17A\u22efCg2 = 167\u00b0 .The mol\u00adecular packing reveals no special supra\u00admolecular features Fig.\u00a02. Most ofI aryl compounds with auxiliary phosphane ligands are relatively rare. According to the CSD database ] -type compounds are 1.922\u2005\u00c5 for the PPh3 derivative 4(PMe3)2] may be attributed to the lower coordination number of the copper atoms. The same effect is also visible for the Cu\u2014P distances of 2.189\u2005\u00c5 2] (R\u2032= Et) with two phosphane units attached to copper. In this case, the copper atom exhibits a distorted trigonal\u2013planar coordination with markedly enlarged Cu\u2014C (1.979\u2005\u00c5) and Cu\u2014P (2.250 and 2.256\u2005\u00c5) distances magnesium units as ligands 4(PMe3)2] (1) was precipitated by the addition of 30\u2005ml of n-hexane. After filtration, the colorless product was washed with diethyl ether (2 \u00d7 5\u2005ml) and dried under vacuum. Single crystals suitable for X-ray analysis were obtained by slow diffusion of n-hexane into a THF solution of the product. Yield: 0.33\u2005g (60%). C42H62Cu4P2 (883.01\u2005g\u2005mol\u22121). Analysis: Cu 29.0% IR (cm\u22121) 2997(w), 2963(m), 2901(m), 2855(w), 2842(w), 2802(w), 2705(w), 1589(w), 1637(w), 1376(w), 1448(w), 1418(m), 1362(w), 1302(w), 1286(m), 1257(w), 1215(w), 1164(w), 1024(w), 939(s), 873(w), 844(s), 730(s), 710(w), 670(m), 576(w), 538(m), 484(w), 357(m), 328(m), 301(m), 275(w). 1H NMR (C6D6): \u03b4 0.60 , 2.08 , 2.73 , 6.68 . 13C{1H} NMR (C6D6): \u03b4 15.4 , 21.4 , 28.7 , 125.6 , 127.4 , 134.4 , 150.0 . 31P{1H} NMR (C6D6): \u03b4 \u221244.6 (s br).A solution of 0.46\u2005g (2.5\u2005mmol) mesitylcopper I. DOI: 10.1107/S2414314621005940/wm4146Isup2.hklStructure factors: contains datablock(s) I. DOI: 2088681CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "It exhibits a defect disk-shaped architecture. DC magnetic property studies in the 2.0\u2013300\u2005K range revealed anti\u00adferromagnetic inter\u00adactions between the CoII ions.A mixed-valence Co 2L) bearing O2N donors for the preparation of a novel Co6 cluster is reported. The hexa\u00adnuclear cobalt com\u00adplex, namely, di-\u03bc2-acetato\u00adtetra\u00adkis\u00ad{\u03bc2-2-[(4-chloro-2-oxido\u00adbenzyl\u00adidene\u00adamino)\u00admeth\u00adyl]phenolato}tetra-\u03bc3-methano\u00adlato-tetra\u00adcobalt(II)dicobalt(III), [CoII4CoIII2(C14H10ClNO2)4(CH3COO)2(CH3O)4], was obtained using Co(CH3COO)2\u00b74H2O and H2L as starting materials in MeOH under solvothermal conditions. The six metal ions are linked together by the \u03bc3-O atoms of four deprotonated MeOH mol\u00adecules, two CH3COO\u2212 units and six phenolate O atoms of four L2\u2212 ligands to form a defect disk-shaped topology. DC magnetic susceptibility investigations revealed the existence of anti\u00adferromagnetic inter\u00adactions in the Co6 cluster.The employment of the new Schiff base ligand 2-[(4-chloro-2-hy\u00addroxy\u00adbenzyl\u00adidene\u00adamino)\u00admeth\u00adyl]phenol (H The tube was sealed and heated at 80 \u00b0C for 48\u2005h under autogenous pressure. It was then cooled to room temperature and dark-red needle-like crystals were obtained. The crystals were collected, washed with MeOH (2\u2005ml) and dried in air . Analysis calculated (%) for C64H58Cl4Co6N4O16: C 47.03, H 3.58, N 3.43; found (%): C 46.18, H 4.048, N 3.280. Selected IR data for 1 (cm\u22121): 1637 (s), 1590 (s), 1523 (s), 1450 (m), 1419 (m), 1286 (w), 1248 (s), 1185 (s), 1089 (m), 1021 (m), 933 (s), 874 (m), 852 (w), 755 (s).To a Pyrex tube (10\u2005ml) was added a mixture of HPLATON = 1.2Ueq(C). The H atoms of CH3 groups were refined as rotating groups, with Uiso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a012L and Co(CH3COO)2\u00b74H2O in MeOH in the presence of NEt3 under solvothermal conditions led to the isolation of 1 in moderate yield. Co(CH3COO)2\u00b74H2O is a good starting material because it not only serves as a convenient metal source, but also provides CH3COO\u2212 bridging ligands. In the solid state, com\u00adplex 1 is stable in air and its elemental analysis is consistent with the given mol\u00adecular formula.The reaction of H1 . The signals of the carb\u00adoxy\u00adl \u03bdas(CO2) and \u03bds(CO2) vibrations were found in the 1637\u20131419\u2005cm\u22121 range. The vibrations of the C=N bond appear at 1450\u2005cm\u22121. Several bands in the 1286\u20131185\u2005cm\u22121 range were assigned to the vibrations of the aromatic rings. The sharp signals in the 979\u2013766\u2005cm\u22121 range were ascribed to the vibrations of C\u2014H bonds.The vibrational bands in the IR spectrum agree well with the formulation of com\u00adplex 1 were obtained from MeOH under solvothermal conditions. Complex 1 crystallized in the ortho\u00adrhom\u00adbic space group P212121. The structure is shown in Fig.\u00a011 is com\u00adposed of six cobalt ions, four 2-[(4-chloro-2-oxido\u00adbenzyl\u00adidene\u00adamino)\u00admeth\u00adyl]phenolate (L2\u2212) ligands, two acetate ligands and four methanol-solvent-derived MeO\u2212 ligands. There exists an approximate C2 symmetry in the mol\u00adecule. The imine N atom and both phenolate O-atom donors of each L2\u2212 ligand coordinate each cobalt centre. Bond valence calculations ]. Of the six cobalt centres, the Co1, Co3, Co4 and Co5 ions are six-coordinated, and the Co2 and Co6 ions are five-coordinated. The coordination environments of the Co2 and Co6 ions, and the Co3 and Co5 ions are individually identical. The Co1 centre is present in a distorted octa\u00adhedral O6 coordination environment, among which two O atoms are from two \u03bc2-\u03ba4O:O,O\u2032,N L2\u2212 ligands and four O atoms are from four \u03bc3-O\u2212 MeO\u2212 ligands. The Co2 centre is enclosed by the N and O atoms of one \u03bc2-\u03ba4O:O,O\u2032,N L2\u2212 ligand, one O atom of a \u03bc3-\u03ba5O:O,N,O\u2032:O\u2032 L2\u2212 ligand and one O atom of one \u03bc3-O\u2212 MeO\u2212 ligand. The six-coordinate NO5 environment around the Co3 ion is accom\u00adplished by two \u03bc3-O\u2212 MeO\u2212 groups, one O atom from one acetate bridge and the N and O atoms of one \u03bc3-\u03ba5O:O,N,O\u2032:O\u2032 L2\u2212 ligand. The six O-donor atoms around the Co6 centre originate from bridging acetate ligands, two \u03bc3-O\u2212 MeO\u2212 groups and two \u03bc3-\u03ba5O:O,N,O\u2032:O\u2032 L2\u2212 ligands. The H2L ligand exhibits two types of coordination mode.Single crystals of SHAPE geometry for both atoms, with a minimum CShM (contunuous shape measure) value of 1.065 for Co2 and 1.172 for Co6.The geometries of the five-coordinated Co2 and Co6 atoms were analyzed with the program 1 joins a small family of Co6 clusters. Hexa\u00adnuclear cobalt com\u00adplexes mainly exhibit wheel, cage and ring topologies /C. The Curie constant C\u00a0= 12.09\u2005cm3\u2005mol\u22121\u2005K and the Weiss constant \u03b8\u00a0= \u221237.24\u2005K were obtained. The negative \u03b8 value proves the anti\u00adferromagnetic inter\u00adactions.Magnetic susceptibility data as a function of temperature for com\u00adplex 1 was also explored. The ac magnetic susceptibilities for 1 at 1000\u2005Hz under a zero-dc field in the temperature range 2\u201325\u2005K were shown in Fig. S2 (see supporting information). The \u03c7\u2032\u2032 susceptibilities at 1000\u2005Hz did not increase upon lowering the temperature and no peaks were determined. These phenomena revealed that com\u00adplex 1 is not a single-mol\u00adecule magnet.The magnetic dynamic behaviour of 2IIICo4II(L)4(CH3COO)2(MeO)4] (1), based on the hydroxy-con\u00adtaining Schiff base ligand 2-[(4-chloro-2-hy\u00addroxy\u00adbenzyl\u00adidene\u00adamino)\u00admeth\u00adyl]phenol (H2L) was prepared and char\u00adacterized. Complex 1 exhibits a defect disk-shaped top\u00adol\u00adogy. Four cobalt ions are six-coordinated and two cobalt ions are five-coordinated. An investigation of the magnetic properties revealed that there exist anti\u00adferromagnetic inter\u00adactions between the CoII ions.A hexa\u00adnuclear cobalt com\u00adplex of com\u00adposition [Co10.1107/S2053229622005885/wv3009sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2053229622005885/wv3009Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2053229622005885/wv3009sup3.pdfIR spectrum, magnetic susceptibility figure, geometry details and table of CShM values. DOI: 1991979CCDC reference:"} +{"text": "I atom exhibits a distorted tetra\u00adhedral coordination environment, defined by three P atoms of three benzyl-di\u00adphenyl\u00adphosphane ligands and one O atom of a nitrato ligand.The central Ag 3)(C19H17P)3], exhibits a severely distorted tetra\u00adhedral coordination environment around the central AgI atom, comprising one O and three P atoms. Apart from a primary Ag\u2014O coordination of the nitrato ligand of 2.667\u2005(3)\u2005\u00c5, a second (weaker) secondary inter\u00adaction of the nitrato ligand via the other O atom of 3.118\u2005(4)\u2005\u00c5 is observed. The compound crystallizes with a complete mol\u00adecule in the asymmetric unit. Weak C\u2014H\u22efO inter\u00adactions consolidate the packing.The mol\u00adecular structure of the title complex, [Ag(NO The nitrato coordinates to the AgI atom via O2 (Ag1\u2014O2 = 2.667\u2005(5)\u2005\u00c5). A secondary weak inter\u00adaction between O1 and Ag1 is also observed, with an inter\u00adaction distance of 3.118\u2005(4)\u2005\u00c5, which is thought to help stabilize the coordination around Ag1. The three N\u2014O bond lengths of the nitrato ligand are nearly identical with a range between 1.221\u2005(3) and 1.233\u2005(3)\u2005\u00c5. The NO3 ligand and the Ag1\u2014P3 bond all lie within the same plane, with P1 and P2 on either side of the plane. Corresponding torsion angles are N1\u2014O2\u2014Ag1\u2014P1 = 110.78\u2005(18)\u00b0, N1\u2014O2\u2014Ag1\u2014P2 = \u2212127.26\u2005(19)\u00b0, and N1\u2014O2\u2014Ag1\u2014P3 = \u22125.9\u2005(2)\u00b0. The concentration of bulky arene groups from the three phosphine ligands also does not appear to notably affect the tetra\u00adhedral environment of each of the P atoms, with typical C\u2032\u2014P\u2014C\u2032\u2032 bond angles between 99.02\u2005(10)\u2013105.80\u2005(11)\u00b0, and an average of 103.40\u00b0.The mol\u00adecular structure of the title compound is shown in Fig.\u00a013 rich layer and an alternating arene-rich layer was added to a solution of silver nitrate (1\u2005mmol) in 20\u2005ml aceto\u00adnitrile. The reaction mixture was heated under reflux for a few hours. It was filtered and left to form crystals. Small colourless crystals were obtained overnight.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314622011476/wm4175sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622011476/wm4175Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622011476/wm4175Isup3.cdxSupporting information file. DOI: 2223250CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title mol\u00adecule is essentially flat. In the crystal the mol\u00adecules are linked by a system of N\u2014H\u22efN hydrogen bonds formed by the hydrazinyl group, a Br\u22efBr halogen bond, and \u03c0-stacking between the pyridine rings. 5H6BrN3, crystallizes in the ortho\u00adrhom\u00adbic space group P212121 with two mol\u00adecules with different conformations in the asymmetric unit. In the crystal, N\u2014H\u22efN and bifurcated N\u2014H\u22ef hydrogen bonds link the mol\u00adecules into [100] chains; a short Br\u22efBr halogen bond and \u03c0\u2013\u03c0 stacking inter\u00adactions are also observed.The title compound, C The asymmetric unit contains two conformationally non-equivalent mol\u00adecules of 6-bromo\u00adpyridin-2-ylhydrazine, (I1) and (I2), as shown in Fig.\u00a01I1) and (I2), respectively. The spatial arrangements of the hydrazino groups, as defined by the torsion angles H2A\u2014N2\u2014N3\u2014H3A = 137\u2005(3)\u00b0 and H5\u2014N5\u2014N6\u2014H6A = 121\u2005(3)\u00b0, correspond to the low-energy conformation that has been calculated for acyl hydrazides and (I2), however. While in (I1) the hydrazine nitro\u00adgen atom N3 is in the syn-disposition with respect to the pyridine nitro\u00adgen atom N1, with N1\u2014C5\u2014N2\u2014N3 = 5.4\u2005(3)\u00b0, in (I2) the hydrazine group is in the anti-conformation, with the corresponding torsion angle N4\u2014C10\u2014N5\u2014N6 = 171.0\u2005(2)\u00b0. For comparison, in 3-chloro\u00adpyrid-2-ylhydrazine for which X-ray diffraction data are available is 2-hydrazino\u00adpyridine; however, no crystal structure of this mol\u00adecule as a free base is known. In crystalline palladium(II) is extensive and involves all nitro\u00adgen atoms of both hydrazine groups and pyridine rings (Table\u00a01I1) and (I2), as shown in Fig.\u00a01A and H3B, and N6 acts as a double acceptor \u2005\u00c5, symmetry code: (i) \u22121\u00a0+\u00a0x, y, z] is about 0.18\u2005\u00c5 shorter than the sum of the van der Waals radii. The aromatic rings of both (I1) and (I2) are involved in a well-defined system of staggered \u03c0\u2013\u03c0 stacking inter\u00adactions \u2005\u00c5], which satisfies the distance and directionality conditions Table\u00a02 for a has Table\u00a03. These vs Table\u00a03.et al., 2004The title compound was prepared following an established synthetic route I. DOI: 10.1107/S2414314623001694/hb4424Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623001694/hb4424Isup3.cmlSupporting information file. DOI: 2243041CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal and mol\u00adecular structure of 2-amino-4-ferrocenyl\u00adthia\u00adzole has been determined. The crystal packing features inter\u00admolecular N\u2014H\u22efN and C\u2014H\u22ef\u03c0 inter\u00adactions. 5H5)(C8H7N2S)], was synthesized by the direct reaction of acetyl\u00adferrocene, thio\u00adurea and resublimed iodine. The structure shows one mol\u00adecule in the asymmetric unit. The amino\u00adthia\u00adzole ring makes an angle of 14.53\u2005(13)\u00b0 with the ferrocenyl ring to which it is attached. In the crystal, pairs of complex mol\u00adecules inter\u00adact via inter\u00admolecular N\u2014H\u22efN hydrogen bonds, forming a cyclic dimer which then inter\u00adacts with other dimers through C\u2014H\u22ef\u03c0 inter\u00adactions.The title compound, [Fe(C The steric effect is also evident in the dihedral angle of 14.77\u2005(17)\u00b0 subtended by the planes of the heterocycle (C14/C15/S11/C12/N13) and the Cp plane (C1\u2013C5).3.Cg(C1\u2013C5) the H-to-ring distance is 2.89\u2005\u00c5, as shown in Table\u00a01bc plane and C\u2014H\u22ef\u03c0 inter\u00adactions. For C10\u2014H10\u22efne Fig.\u00a02. The str4.et al., 2016et al., 2005et al., 2007et al., 2006bet al., 2006aet al., 2005et al., 2006aet al., 2020et al., 20052sp hybridization for all C and N atoms.A search of the Cambridge Structural Database \u03bd 3099 (ArCH), 2921 (CH3), 1658 (C=N); 1H NMR : 4.62 ; 4.25 ; 4.10 ; 5.00 , 6.35 .The title compound was synthesized according to the reported method (Chopra 6.Uiso(H) = 1.2Ueq(N). C-bound H atoms were positioned geometrically (C\u2014H = 0.93\u20130.98\u2005\u00c5) and refined with isotropically Uiso(H) = 1.2Ueq(C) using a riding model.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022007228/dj2046sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022007228/dj2046Isup3.hklStructure factors: contains datablock(s) I. DOI: 1841501CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the salicyl\u00adaldehyde alcohol group is engaged in an intra\u00admolecular O\u2014H\u22efN hydrogen bond with the imine nitro\u00adgen atom, while the tertiary alcohol is engaged in a weak inter\u00admolecular O\u2014H\u22efF hydrogen bond with an adjacent mol\u00adecule. 28H24FNO2, crystallizes in the ortho\u00adrhom\u00adbic space group P212121. A hydrogen-bonding network between the tertiary alcohol group and the fluoro substituent results in [010] chains in the solid state.The title compound, C S)- or (R)-2-amino-1,1,3-tri\u00adphenyl\u00adpropanol imine, C28H24FNO2, crystallizes in the ortho\u00adrhom\u00adbic space group P212121 as shown in Fig.\u00a01l-enanti\u00adomer structure was collected at 100\u2005K while the d-enanti\u00adomer was collected at 293\u2005K. The unit-cell parameters in the current room-temperature structure are slightly larger (average 1.3%), presumably due to the higher temperature of the data collection. The absolute structure parameter of \u22120.1\u2005(3) has a large uncertainty but the absolute configuration was verified by synthesis and polarimetry.2-Hy\u00addroxy-5-fluoro-benzaldehyde 2The compound has the expected imine\u2013phenol structure as opposed to the iminium\u2013phenoxide tautomer seen in derivatives with less steric bulk. The C23\u2013C28 phenol aromatic ring is close to co-planar with atoms O2 [deviation from the ring plane = 0.040\u2005(2)\u2005\u00c5], C22 [\u20130.061\u2005(2)\u2005\u00c5], N1 [\u20130.034\u2005(2)\u2005\u00c5] and C2 [\u20130.039\u2005(2)\u2005\u00c5]. These four atoms exhibit less deviation from the plane than the enanti\u00adomer. The C22\u2014N1\u2014C2\u2014C1 torsion angle is 110.2\u2005(2)\u00b0, which places atom O1 1.555\u2005(2)\u2005\u00c5 above the plane of the ring. This deviation is 0.166\u2005\u00c5 larger than that for the enanti\u00adomer at 100\u2005K, although the torsion angle is almost identical.et al., 2019The bonds between C27\u2014C28, C23\u2014C28 and C23\u2014C24 are long at 1.39\u20131.41\u2005\u00c5 while those between C24\u2014C25, C25\u2014C26 and C26\u2014C27 are shorter at 1.36\u20131.37\u2005\u00c5. In contrast, the aromatic rings on the benzyl and phenyl substituents have typical C\u2014C bond distances ranging from 1.37\u20131.39\u2005\u00c5. The aromatic C28\u2014O2 bond at 1.349\u2005(3)\u2005\u00c5 is substanti\u00adally shorter than the aliphatic C1\u2014O1 bond [1.439\u2005(3)\u2005\u00c5]. This bonding motif has been seen in related structures (Sha S(6) ring and a long-range inter\u00admolecular hydrogen bond between the tertiary alcohol O1\u2014H1 and the F1 atom of an adjacent mol\u00adecule as shown in Fig.\u00a02There is an intra\u00admolecular O2\u2014H2\u22efN1 hydrogen bond Table\u00a01 between et al., 2019Preparative details of the material have been reported previously global, I. DOI: 10.1107/S2414314620015801/hb4369Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314620015801/hb4369Isup3.cmlSupporting information file. DOI: 1970566CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Zr\u2010bptc shows a high SO2 uptake of 6.2\u2005mmol\u2009g\u22121 at 0.1\u2005bar and 298\u2005K, reflecting excellent capture capability and removal of SO2 at low concentration (2500\u2005ppm). Dynamic breakthrough experiments confirm that the introduction of amine, atomically\u2010dispersed CuII or heteroatomic sulphur sites into the pores enhance the capture of SO2 at low concentrations. The captured SO2 can be converted quantitatively to a pharmaceutical intermediate, aryl N\u2010aminosulfonamide, thus converting waste to chemical values. In situ X\u2010ray diffraction, infrared micro\u2010spectroscopy and inelastic neutron scattering enable the visualisation of the binding domains of adsorbed SO2 molecules and host\u2013guest binding dynamics in these materials at the atomic level. Refinement of the pore environment plays a critical role in designing efficient sorbent materials.We report reversible high capacity adsorption of SO 2 uptake. The incorporation of atomically\u2010dispersed CuII, amine or heteroatomic sulphur sites enhances the uptake of SO2 at low concentrations. This work confirms that control of pore environments is an important approach for optimising the adsorption of SO2 at low concentrations.A series of Zr\u2010based metal\u2013organic framework materials have been investigated for reversible SO MOFs constructed from {Zr6} clusters are renowned for their high stability. and Zr\u2010DMTDC6(\u03bc3\u2010O)4(\u03bc3\u2010OH)4(OOCR)12} clusters bridged by dicarboxylates to give cubic structures of fcu topology 4(\u03bc3\u2010OH)4(OOCR)12} clusters and tetracarboxylate ligands in an open framework of ftw topology.2\u2009g\u22121. MFM\u20101336(OH)8(OH)8(OOCR)8} clusters and thcb4\u2212 ligands \u20102,2\u2032,4,4\u2032,6,6\u2032\u2010hexamethyl\u20101,1\u2032\u2010biphenyl) to form a flu topology. MFM\u2010133 shows an axially elongated octahedral cage (10.4\u00d710.4\u00d725.9\u2005\u00c5) and a BET surface area of 2156\u2005m2\u2009g\u22121 8(OH)8(OOCR)8} clusters with the tetratopic ligand 3,3\u2032\u2032,5,5\u2032\u2032\u2010tetrakis(4\u2010carboxyphenyl)\u2010p\u2010terphenyl (H4tcpt) to give a neutral framework of sqc topology. MFM\u2010422 is comprised of a trigonal cage and a hexagonal cage .Zr\u2010bptc is built from 12\u2010connected {Zr2 have been recorded for these MOFs at 273\u2013298\u2005K and from 0\u20131\u2005bar (36.7\u2005mmol\u2009g\u22121) under the same conditions.2, UiO\u201066\u2010CuII, Zr\u2010DMTDC, Zr\u2010bptc, MFM\u2010133 and MFM\u2010422, show fully reversible uptakes of SO2 of 8.6, 8.8, 8.2, 9.6, 7.8, 8.9 and 13.6\u2005mmol\u2009g\u22121, respectively . The comparable adsorption uptakes of UiO\u201066, UiO\u201066\u2010NH2 and UiO\u201066\u2010CuII at 1\u2005bar (8.2\u20138.8\u2005mmol\u2009g\u22121) suggest that decoration of the pore environment with functional groups or open CuII sites has little impact on the total uptake capacity, which is determined primarily by the surface area. The slightly higher uptake of Zr\u2010DMTDC (9.6\u2005mmol\u2009g\u22121) is consistent with its higher surface area (1345\u2005m2\u2009g\u22121), compared with the other three UiO\u201066 materials. In contrast, enhancements in the uptake at 0.1\u2005bar were observed for UiO\u201066\u2010NH2, UiO\u201066\u2010CuII and Zr\u2010DMTDC, compared with UiO\u201066 for SO2 uptake show decreasing values of 45\u201350, 44\u201332, 38\u201334, 32\u201329, 37\u201327, 31\u201327 and 26\u201319\u2005kJ\u2009mol\u22121 for Zr\u2010bptc, UiO\u201066\u2010NH2, UiO\u201066\u2010CuII, Zr\u2010DMTDC, UiO\u201066, MFM\u2010422 and MFM\u2010133, respectively. Compared with UiO\u201066, the materials UiO\u201066\u2010NH2, UiO\u201066\u2010CuII and Zr\u2010DMTDC show higher values for Qst, consistent with the enhanced adsorption at low pressure. The relatively low values of Qst for MFM\u2010133 and MFM\u2010422 are consistent with their large pores, reducing the strength of host\u2013guest interactions.Gravimetric adsorption isotherms of SO2 and N2 have also been recorded for Zr\u2010bptc, UiO\u201066\u2010NH2, UiO\u201066\u2010CuII, Zr\u2010DMTDC and UiO\u201066 to assess the adsorption selectivity 2/CO2 (1/99) and SO2/N2 (1/99) at 298\u2005K and 1.0\u2005bar. UiO\u201066, UiO\u201066\u2010NH2, Zr\u2010DMTDC and UiO\u201066\u2010CuII exhibit retention times for SO2 in the expected order of 33, 53, 58 and 100\u2005min\u2009g\u22121, respectively with retention times of 80, 175, 157 and 175\u2005min\u2009g\u22121 for UiO\u201066, UiO\u201066\u2010NH2, Zr\u2010DMTDC and UiO\u201066\u2010CuII, respectively and SO2/N2 (2500\u2005ppm SO2/75\u2009% CO2 in He), respectively 4(bdc)6\u2009\u22c5\u2009(SO2)7.7] reveal two binding sites I and II located in cage T (SO2/{Zr6}=5.1) and cage O (SO2/{Zr6}=2.6), respectively \u2005\u00c5] and dipole\u2013dipole interaction [O2S\u22c5\u22c5\u22c5phenyl ring=3.69(2)\u2005\u00c5] stabilise SO2 (I) is stabilised by two hydrogen bonds 4(bdc\u2212NH2)6\u2009\u22c5\u2009(SO2)8.1], two binding sites I\u2032 and II\u2032 are observed in cage T (SO2/{Zr6}=4.7) and cage O (SO2/{Zr6}=3.4), respectively \u2005\u00c5] was identified and works together with an interaction [O2S\u22c5\u22c5\u22c5phenyl ring=3.58(1)\u2005\u00c5] and hydrogen bonding [OSO\u22c5\u22c5\u22c5\u03bc3\u2010HO=2.94(5)\u2005\u00c5] that stabilise SO2 binding at site I\u2032 \u2005\u00c5, SO2\u22c5\u22c5\u22c5NH2=1.73(3), 2.43(6), 2.87(7), 3.21(1), 3.30(3) and 3.63(8)\u2005\u00c5], which work together with two further dipole\u2013dipole interactions [O2S\u22c5\u22c5\u22c5NH2=2.40(4) and 3.10(7)\u2005\u00c5] to stabilise SO2 at site II\u2032 4(DMTDC)2\u2009\u22c5\u2009(SO2)13.1], four binding sites were revealed (I\u2032\u2032\u2010IV\u2032\u2032). Sites I\u2032\u2032, II\u2032\u2032 and III\u2032\u2032 are localised in cage T and 3.52(7)\u2005\u00c5; O2S\u22c5\u22c5\u22c5thiophene ring=3.65(3)\u2005\u00c5] and 2.70(4)\u2005\u00c5; O2S\u22c5\u22c5\u22c5thiophene ring=3.72(3) and 3.72(3)\u2005\u00c5] and supramolecular interaction [O2S\u22c5\u22c5\u22c5\u03bc3\u2010O=3.63(9)\u2005\u00c5]. In addition, dipole\u2013dipole interaction between SO2 at sites I\u2032\u2032 and II\u2032\u2032 [OSO(I\u2032\u2032)\u22c5\u22c5\u22c5SO2(II\u2032\u2032)=2.81(7)\u2005\u00c5] was identified \u2005\u00c5] and [OSO\u22c5\u22c5\u22c5S\u2010ring=3.97(6)\u2005\u00c5] were identified between SO2(III\u2032\u2032) and the framework and 3.77(8)\u2005\u00c5] and a hydrogen bond [OSO\u22c5\u22c5\u22c5H3C=2.42(2)\u2005\u00c5] 4(bptc)3\u2009\u22c5\u2009(SO2)5.8], six binding sites were revealed (I\u2013VI) \u2005\u00c5] and by two hydrogen bonds [OSO\u22c5\u22c5\u22c5H\u2212C=2.79(8)\u2005\u00c5 and OSO\u22c5\u22c5\u22c5\u03bc3\u2010OH=2.36(5)\u2005\u00c5] \u22c5\u22c5\u22c5SO2(I)=2.06(8) and 3.07(2)\u2005\u00c5, OSO(I)\u22c5\u22c5\u22c5SO2(II)=3.28(1)\u2005\u00c5] with SO2 at site I \u22c5\u22c5\u22c5OSO(V)=2.99(7)\u2005\u00c5, OSO(III)\u22c5\u22c5\u22c5SO2(V)=3.13(5)\u2005\u00c5] with SO2 at site V immobilised by dipole\u2013dipole interactions [O2S\u22c5\u22c5\u22c5phenyl ring=3.72(5)\u2005\u00c5] and two\u2010fold electrostatic interactions [OSO\u22c5\u22c5\u22c5H\u2212C=2.78(1) and 2.93(9)\u2005\u00c5] \u2005\u00c5] and dipole\u2013dipole interaction [OSO(IV)\u22c5\u22c5\u22c5SO2(VI)=3.83(5)\u2005\u00c5] with SO2 at site VI , 1.88(6), 2.23(5), 2.23(5) and 2.69(7)\u2005\u00c5] and two dipole\u2013dipole interactions [O2S\u22c5\u22c5\u22c5OOC=3.07(2) and 3.07(2)\u2005\u00c5] and carboxylic groups and multiple strong hydrogen bonding at site VI jointly facilitate the exceptional SO2 uptake at low pressure.In SO2 (0\u20131\u2005bar) in the UiO\u201066 type systems have been analysed by in situ synchrotron infrared micro\u2010spectroscopy. For all the MOFs, clear binding of SO2 to the hydroxyl group is observed with a red shift of the \u2212OH stretching vibration at \u22483671\u2005cm\u22121 by 86, 95, 83 and 82\u2005cm\u22121 in UiO\u201066, UiO\u201066\u2010NH2, UiO\u201066\u2010CuII and Zr\u2010DMTDC, respectively mode was monitored to examine the displacement of bound CO2 by SO2 stretch corresponding to the bare and CO2\u2010loaded materials are approximately equal. Due to weak interaction between CO2 and the \u03bc3\u2010OH group, the bare \u03bc3\u2010OH band is not fully depleted but a new peak at 3643\u2005cm\u22121 appears and is assigned to the [OH\u22c5\u22c5\u22c5OCO] band , there is a steady change in the \u03bd(\u03bc3\u2010OH) region that includes new bands appearing in a similar manner to the pure SO2 experiment, indicating that bound CO2 does not impede SO2 adsorption . Upon 30\u2009% SO2\u2010loading, the characteristic [OH\u22c5\u22c5\u22c5OCO] band has fully disappeared showing that SO2 readily displaces bound CO2 in the pore as a result of stronger binding. Hence, selective capture of SO2 from a mixture of SO2/CO2 can be achieved as demonstrated in separation experiments. Furthermore, 40\u2009%, 45\u2009% and 50\u2009% SO2\u2010loadings fully displace CO2 in UiO\u201066\u2010NH2, Zr\u2010DMTDC and UiO\u201066, respectively . The competitive binding studies of SO2/CO2 further confirm enhanced SO2 binding in the decorated MOFs. The decreasing partial pressure of SO2 on full displacement of CO2 in UiO\u201066\u2010CuII, UiO\u201066\u2010NH2 and Zr\u2010DMTDC is consistent with that observed in static and dynamic adsorption studies.Upon stepwise dosing of the COIn situ INS, coupled with DFT calculations, enables the visualization of binding dynamics for SO2\u2010loaded Zr\u2010bptc. Seven major changes in the INS spectra were observed on the adsorption of SO2 in Zr\u2010bptc was observed at 0.1\u2005bar and 298\u2005K. Furthermore, the captured SO2 in Zr\u2010bptc can be converted readily into fine chemicals, paving new pathways to \u201cwaste\u2010to\u2010chemicals\u201d technologies. In situ SXPD, microFTIR and INS studies, coupled with DFT calculations, unravel the molecular details of host\u2013guest binding that result in the enhancement of SO2 adsorption at low pressure in these materials. These studies confirm that control of pore environments is an important approach for improving the adsorption of SO2.Powerful drivers exist for the development of new regenerable sorbents for SO6(OH)8(OH)8(tcpt)2], [Zr6O4(OH)4(bptc)3\u2009\u22c5\u2009(SO2)5.8], [Zr6O4(OH)4(DMTDC)6\u2009\u22c5\u2009(SO2)13.1], [Zr6O4(OH)4(bdc)6\u2009\u22c5\u2009(SO2)7.7] and [Zr6O4(OH)4(bdc\u2212NH2)6\u2009\u22c5\u2009(SO2)8.1] are available free of charge from the Cambridge Crystallographic Data Centre .Additional crystallographic information, gas adsorption data, thermogravimetric analysis, density function theory (DFT) calculations and breakthrough data are available in the Supporting Information. The crystal structures of [ZrThe authors declare no conflict of interest.1As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file.Supporting InformationClick here for additional data file.Supporting InformationClick here for additional data file.Supporting InformationClick here for additional data file.Supporting InformationClick here for additional data file."} +{"text": "In the crystal, the mol\u00adecules are linked by O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds into a three-dimensional network. 6H8NO+\u00b7C2H3O2\u2212, the cations and anions are linked by O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, generating a three-dimensional network.In the title molecular salt, C The C\u2014OH bond length (C2\u2014O1) of 1.3520\u2005(9)\u2005\u00c5 is similar to that observed for structures containing 2-hy\u00addroxy\u00adbenzenaminium as a cation [1.350\u2005(3)\u2005\u00c5; Jin & Wang, 2013C\u22efO3 cation\u2013anion hydrogen bonds generate a succession of infinite chains [graph set c-axis direction and acetic acid. This mixture was obtained by dissolution and agitation under reflux for 3\u2005h of 0.5\u2005g of the 2-amino\u00adphenol and 0.27\u2005g of acetic acid in a 1:1 stoichiometric ratio in a hot ethano\u00adlic solution (20\u2005ml). After warming for a few minutes using a water bath, the solution was cooled and kept at room temperature. Within a few days, yellow needle-like crystals suitable for the X-ray analysis were obtained (yield 60%) by evaporation of the solution.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314622001122/vm4050sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622001122/vm4050Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622001122/vm4050Isup3.cmlSupporting information file. DOI: 2149479CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The plane resulting from the NO2 coordination to Ag is nearly perpendicular to the plane from the coordination of the phosphine-P atoms to Ag [dihedral angle = 86.43\u2005(9)\u00b0].The title Ag P21/c with Z = 4. The asymmetric unit contains one complete silver complex mol\u00adecule, featuring an AgI atom, two diphenyl-p-tolyl\u00adphosphine ligands, and one NO2 coordinating in a bidentate fashion. Near-identical Ag\u2014P bond lengths are observed [Ag1\u2014P1 = 2.4209\u2005(7)\u2005\u00c5 and Ag1\u2014P2 = 2.4251\u2005(8)\u2005\u00c5]. The nitrito ligand is similarly coordinating in a near symmetric fashion (Ag1\u2014O1 = 2.422\u2005(2), Ag1\u2014O2 = 2.415\u2005(2), N1\u2014O1 = 1.253\u2005(4) and N1\u2013O2 = 1.255\u2005(4)\u2005\u00c5). As seen in Fig.\u00a01ipso-carbon atoms of each of the phosphine ligands overlap in a near-eclipsed fashion when viewed down the P1\u2014Ag1\u2014P2 plane Pl2. Corresponding torsion angles are Ag1\u2014P1\u2014C1\u2014C2 = \u221223.4\u2005(3)\u00b0, Ag1\u2014P1\u2014C7\u2014C8 = \u221251.9\u2005(3)\u00b0, Ag1\u2014P1\u2014C13\u2014C14 = 147.8\u2005(3)\u00b0, Ag1\u2014P2\u2014C20\u2014C21 = \u221229.0\u2005(3)\u00b0, Ag1\u2014P2\u2014C26\u2014C27 = 133.3\u2005(3) and Ag1\u2014P2\u2014C32\u2014C33 = 132.3\u2005(3)\u00b0. The complex packs in three dimensions as layers of mol\u00adecules, leaving thin corrugated channels in between the inorganic layers when viewed along the a axis was dissolved in aceto\u00adnitrile (10\u2005ml). Silver nitrite (1\u2005mmol) was dissolved in aceto\u00adnitrile (5\u2005ml). The diphenyl-p-tolyl\u00adphosphine solution (10\u2005ml) was added to the silver nitrite solution (5\u2005ml), to give a 2:1 molar ratio reaction. The mixture was heated under reflux for 2\u2005h after which the solution was left to crystallize.Diphenyl-For full experimental details including crystal data, data collection and structure refinement details, refer to Table\u00a0110.1107/S2414314622007714/tk4082sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622007714/tk4082Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622007714/tk4082Isup3.cdxSupporting information file. DOI: 2193913CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, weak C\u2014H\u22efCl/O/\u03c0 inter\u00adactions are observed.The title compound, [RuCl 2(C10H14)(C21H21O3P)], crystallizes with two complex mol\u00adecules in the asymmetric unit. The RuII atom has a classical three-legged piano-stool environment being coordinated by a cymene ligand [Ru\u2014centroid = 1.707\u2005(2)/1.704\u2005(2)\u2005\u00c5], a tris\u00ad(4-meth\u00adoxy\u00adphen\u00adyl)phosphane ligand [Ru\u2014P = 2.3629\u2005(15)/2.3665\u2005(15)\u2005\u00c5] and two chloride atoms with the Ru\u2014Cl bonds adopting two distinct values of 2.4068\u2005(16)/2.4167\u2005(16) and 2.4016\u2005(15)/2.4244\u2005(16)\u2005\u00c5. The effective cone and solid angles for the phosphane ligands were calculated to be 149.5/150.2\u00b0 and 25.3/25.6\u00b0, respectively. In the crystal, weak C\u2014H\u22efCl/O/\u03c0 inter\u00adactions are observed. The crystal was refined as a two-component twin.The title compound, [RuCl II\u2013arene complexes is well known in the catalytic transfer hydrogenation of carbonyl compounds 3, as part of ongoing structural investigations into these type of complexes.The activity of the half-sandwich RuPZ = 4), with its two unique mol\u00adecules adopting a distorted pseudo-octa\u00adhedral arrangement, revealing the typical three-legged piano-stool geometry. The coordination sphere of the ruthenium is occupied by a cymene, a tris\u00ad(4-meth\u00adoxy\u00adphen\u00adyl)phosphane and two chloride atoms and 1.704\u2005(2)\u2005\u00c5 for the two independent mol\u00adecules; the mean Ru\u2014C bond lengths are 2.217\u2005(6) and 2.214\u2005(6)\u2005\u00c5. The coordination of the remaining ligands to the Ru atom shows a slight deviation from the typical octa\u00adhedral geometry with Cl\u2014Ru\u2014Cl = 88.47\u2005(6) and 88.77\u2005(6)\u00b0, respectively; Cl\u2014Ru\u2014P = 86.50\u2005(5)/88.03\u2005(5) and 86.05\u2005(5)/88.21\u2005(6)\u00b0. The Ru\u2014P bond lengths are 2.3629\u2005(15) and 2.3665\u2005(15)\u2005\u00c5, while the Ru\u2014Cl bonds adopt two distinct values of 2.4068\u2005(16)/2.4167\u2005(16) and 2.4016\u2005(15)/2.4244\u2005(16)\u2005\u00c5 for Ru1 and Ru2, respectively. The above bond lengths are within normal ranges as data extracted from the Cambridge Structural Database RuCl2(PR3) systems from 429 hits, containing 535 usable Ru\u2014Cl observations, show a mean value of 2.412\u2005(12)\u2005\u00c5 in a range from 2.378 to 2.459\u2005\u00c5. The same group of structures show for the Ru\u2014P distance a mean value of 2.34\u2005(3)\u2005\u00c5 in a range from 2.235 to 2.434\u2005\u00c5. The geometries of the two independent mol\u00adecules are virtually identical, as seen from a superimposed fit with an r.m.s. deviation of 0.0525\u2005\u00c5 3 in CH2Cl2 (10\u2005ml) was added to a stirred orange solution of [Ru(p-cymene)Cl2]2 under Ar in the same solvent (15\u2005ml) and stirred at r.t. for 24\u2005h. The resulting red reaction mixture was filtered, the filtrate concentrated under reduced pressure to ca 5\u2005ml. Cold diethyl ether (10\u2005ml) was carefully added and the solvent left to slowly evaporate whereby a sample of [RuCl2(C10H14)(C21H21O3P)] suitable for single-crystal X-ray diffraction was obtained as orange crystals.A solution of P: \u03b4 (p.p.m.) 21.39 (s). 1H NMR : \u03b4 (p.p.m.) 1.11 ; 1.84 ; 2.87 ; 3.78 ; 4.93 ; 5.20 ; 6.85 ; 7.69 .Analytical data: \u22123) is located at 0.59\u2005\u00c5 from Ru1 and the highest peak (3.95\u2005e\u2005\u00c5\u22123) 0.90\u2005\u00c5 from Ru1. Initial refinement of data indicated a two-component twin with a 180\u00b0 rotation about the [100] reciprocal direction. Refinement with the appropriate twin law yields a batch scaling factor of 0.18.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621012591/tk4072sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314621012591/tk4072Isup2.hklStructure factors: contains datablock(s) I. DOI: 2124507CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal features 4 16H13ClN2O3)(C3H4N2)], the NiII ion is coordinated by two O atoms and one N atom derived from the dianionic N\u2032-[(1E)-1-(5-chloro-2-hy\u00addroxy\u00adphen\u00adyl)ethyl\u00adidene]-4-meth\u00adoxy\u00adbenzohydrazide ligand and one N atom from the imidazole mol\u00adecule. The N2O2 donor set defines an approximate square-planar geometry. The dihedral angles between the imidazole ring and the fused six-membered and meth\u00adoxy\u00adbenzene rings are 17.78\u2005(14) and 13.23\u2005(16)\u00b0, respectively; the dihedral angle between the C6 rings is 6.63\u2005(12)\u00b0. The most prominent feature of the mol\u00adecular packing is the formation of 41-helical chains mediated by imidazole-N\u2014H\u22efO(phenoxide) hydrogen bonding; these are linked by methyl-C\u2014H\u22efCl inter\u00adactions.In the title complex, [Ni(C In this complex, the Ni atom is located in a slightly distorted square-planar environment and C10\u2013C15 (B), and the imidazole ring (C) make dihedral angles of 6.63\u2005(12)\u00b0 (A/B), 17.78\u2005(14)\u00b0 (A/C) and 13.23\u2005(16)\u00b0 (B/C).The Nint Fig.\u00a01. The Ni\u2014c axis, which leads to a 41 helical chain. The chains are connected by C\u2014H\u22efCl inter\u00adactions (Table\u00a02The mol\u00adecular packing is consolidated by imidazole-N\u2014H\u22efO(phenoxide) hydrogen bonding Table\u00a02 along ths Table\u00a02 into a tN\u2032-[(1E)-1-(5-chloro-2-hy\u00addroxyphen\u00adyl)ethyl\u00adidene]-4-meth\u00adoxy\u00adbenzohydrazide , 1H-imidazole , Ni(NO3)2\u00b76H2O , methanol (10\u2005ml) and distilled water (5\u2005ml) were mixed in a 50\u2005ml flask. The mixture was stirred at room temperature for 1\u2005h, the pH was adjusted with saturated sodium carbonate solution to about 8 followed by filtration. Red rectangular block-shaped crystals were obtained after about one month by evaporating the filtrate in air (yield 31%).The Schiff base ligand, Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2414314622002954/tk4076sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622002954/tk4076Isup2.hklStructure factors: contains datablock(s) I. DOI: 2159183CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "STING is an innate immune sensor for immune surveillance of viral/bacterial infection and maintenance of an immune\u2010friendly microenvironment to prevent tumorigenesis. However, if and how STING exerts innate immunity\u2010independent function remains elusive. Here, the authors\u00a0report that STING expression is increased in renal cell carcinoma (RCC) patients and governs tumor growth through non\u2010canonical innate immune signaling involving mitochondrial ROS maintenance and calcium homeostasis. Mitochondrial voltage\u2010dependent anion channel VDAC2 is identified as a new STING binding partner. STING depletion potentiates VDAC2/GRP75\u2010mediated MERC (mitochondria\u2010ER contact) formation to increase mitochondrial ROS/calcium levels, impairs mitochondria function, and suppresses mTORC1/S6K signaling leading to RCC growth retardation. STING interaction with VDAC2 occurs through STING\u2010C88/C91 palmitoylation and inhibiting STING palmitoyl\u2010transferases ZDHHCs by 2\u2010BP significantly impedes RCC cell growth alone or in combination with sorafenib. Together, these studies reveal an innate immunity\u2010independent function of STING in regulating mitochondrial function and growth in RCC, providing a rationale to target the STING/VDAC2 interaction in treating RCC. STING is a canonical innate immune sensor triggering type\u2010I interferon production. Here, an innate immunity\u2010independent STING function in governing renal cancer cell (RCC) proliferation is reported through regulating mitochondria calcium homeostasis by binding mitochondrial calcium transporter VDAC2, as well as its therapeutic potential in treating RCC. Viral and bacterial DNA, as well as damaged genomic and mitochondria DNA, are sensed by the cytosolic DNA sensor cGAS (cyclic GMP\u2010AMP\u00a0synthase). Binding of DNA to cGAS activates its enzymatic activity by triggering cGAS dimerization and phase transition toward synthesizing a special unsymmetric cyclic dinucleotide 2\u20323\u2032\u2010cGAMP. Subsequently, 2\u20323\u2032\u2010cGAMP binds STING (stimulator of interferon response cGAMP interactr 1) on the endoplasmic reticulum (ER) to promote its trafficking to Golgi and other peri\u2010nuclear membrane\u2010coated compartments, where STING recruits TBK1 to phosphorylate IRF3. Phosphorylated IRF3 dimerizes and translocates into nuclei in triggering the expression of interferon \u03b2 (IFN\u03b2). This leads to expression of immune modulatory genes and activation of adaptive immunity. However, STING can also promote autophagy, lysosome\u2010mediated cell death of myeloid cells, and inhibition of viral replication via IFN\u03b2 \u2010independent mechanisms. In addition, STING also plays a critical role in responding to RNA viruses including influenza A viruses (IAV), vesicular stomatitis virus (VSV), dengue virus (DENV) and others, through directly binding and modulating function of essential RNA sensors RIG\u2010I and MAVS and other mechanisms.Innate immunity serves as the first line of defense against infectious pathogens in vertebrates to restrain pathogen invasion and activate adaptive immunity. Pathogen\u2010associated molecular patterns (PAMPs) help distinguish between self\u2010 and non\u2010self\u2010materials and initiate recognition and activation of innate immunity.1 Nucl) and genetic animal models . Moreover, evidence from gt/gtSting (Sting knockout) mice suggests that activation of the STING signaling creates an immune\u2010responsive microenvironment that suppresses tumorigenesis, and STING loss has been observed in colon cancer, gastric cancer and melanoma and restrains IFN\u2010mediated tissue repair and T cell priming. Activating STING has been shown to improve therapeutic effects of immune\u2010checkpoint blockades. STING also exerts a protective effect in metabolic diseases such as alcoholic liver disease, cardiovascular diseases including myocardial infarction, aging, and others through promoting IRF3 phosphorylation and activation of innate immune signaling.Other than nucleotide sensing, hyperactivation of STING has been observed and connected with inflammatory diseases from both pathological perspectives disease is caused by GOF mutations in STING18) an\u03b2 \u2010independent T cell death by inducing ER stress in T cells. STING depletion has been shown to promote cancer cell growth via accelerated S\u2010phase entry and increased chromosome instability in the absence of an immune environment. Interestingly, STING activation can mediate mitochondrial damage that leads to renal fibrosis and renal failure. Although extensive studies demonstrate that STING is involved in controlling tumorigenesis and renal failure, there is no evidence of STING involvement in the development of RCC, especially through a non\u2010canonical and innate immunity\u2010independent manner.Innate immunity\u2010independent STING function has also been investigated but is less understood. To this end, although STING activation mediates T cell tumor infiltration, STING was also reported to mediate IFN T cells.29 STI22.1 Our analyses of the TCGA dataset identified gene amplification of STING but not cGAS (MB21D1) showed STING overexpression.33 Our) Figure\u00a0. Further cGAS MB21 Figure\u00a0. Depleti) Figure\u00a0 led to r) Figure\u00a0 and decr) Figure\u00a0 with the) Figure\u00a0. In cont) Figure\u00a0 also red) Figure\u00a0. We next2.2STING\u2010S366A knockin A498 cells by CRISPR and mitochondrial ROS (by MitoSOX Red), but also mitochondrial calcium levels (by Rhod\u20102 AM) Figure\u00a0 were incA Figure\u00a0, suggest2.4 ATP2A2 (SERCA Ca\u2010ATPase located on ER membrane), and ATP1A1 . Binding of STING with VDAC2, ATP2A2, and ATP1A1 was confirmed in both A498 and mitochondrial calcium uniporters (such as VDACs). In this process, GRP75 (glucose\u2010regulated protein 75) at the mitochondria\u2010associated membrane (MAM) bridges IR3R interaction with VDACs. Given we observed STING depletion induced increased Ca2+ flux into mitochondria and upregulated mito\u2010ROS levels and ECAR (extracellular acidification rate) by seahorse. STING\u2010depleted A498 cells displayed a decreased basal respiration rate compared with control cells and a decreased ATP\u2010driven synthesis ability Figure\u00a0 may caus\u03b2L, Raptor, and Sin1 could promote mTORC1/S6K activation in A498 cells would be an approach to maximize tumor inhibition. In addition, considering that VDAC2 cysteine residue palmitoylation and oxidation were also previously detected by mass spectrometry, we analyzed if any of these cysteine mutations may affect VDAC2 binding to STING. We generated VDAC2\u2010C103A, C210A, C227A, and C103A/C210A/C227A (3A) mutants and found none of these mutants significantly affect VDAC2 binding to STING , we depleted the protein acyl transferases ZDHHC3 or ZDHHC7 using shRNAs Figure\u00a0, mimicki65 2\u2010Bs Figure\u00a0 and STINs Figure\u00a0, 2\u2010BP trs Figure\u00a0 to facils Figure\u00a0. Notablys Figure\u00a0 through s Figure\u00a0 as STINGs Figure\u00a0. Like STs Figure\u00a0, 2\u2010BP trs Figure\u00a0. As a res Figure\u00a0, while ds Figure\u00a0. Moreoves Figure\u00a0. Interess Figure\u00a0. Like STs Figure\u00a0, 2\u2010BP tr2.8 and currently various targeted therapies have been approved in treating RCC patients in clinic including tyrosine kinase inhibitors , HIF inhibitors (belzutifan) and mTOR inhibitors (temsirolimus and everolimus), we next examined if 2\u2010BP exerts any synergistic effects with clinically approved agents. To this end, although 2\u2010BP reduced mTORC1/S6K activation in RCC cells and THCA (thyroid carcinoma). If STING is also a vulnerability in these two types of cancers or if STING regulates mitochondria ROS/calcium homeostasis in STAD and THCA remains an interesting topic for further investigations.It remains unclear why unlike other cancers including colon cancer,21 gasTMEM173 (STING) gene is observed in different ethnic populations\u2010 for example, HAQ (R71H\u2010G230A\u2010R293Q) is dominant in East Asian, while Africans have no HAQ. If these various STING variants affect STING binding with VDAC2 and determine STING dependency for cell growth in cancers beyond RCC warrants further in\u2010depth investigations.Notably, the heterogeneity of e no HAQ.69 If 4l\u2010Glutathione (reduced) was purchased from Gold Biotechnology (G\u2010155\u201025). 4\u2010hydroxy Nonenal (4\u2010HNE) (32100) was purchased from Cayman. Sulfobutylether\u2010\u03b2\u2010Cyclodextrin (HY\u201017031), Sorafenib (HY\u201010201A), and RSL3 (HY\u2010100218A) were purchased from MedChemExpress. Everolimus (A8169) was purchased from ApexBio Technology. Temsirolimus (T3574) was purchased from TCI. H\u2010151 (6675) was purchased from Tocris.Bafilomycin A1 (S1413), cycloheximide (S6611), erastin (S7242), MG132 (S2619), MLN4924 (S7109), S6K1\u2010I (S2163), tBHQ (S4990) and rapamycin (AY\u201022989) were purchased from Selleck. 2\u2010bromohexadecanoic acid (2\u2010BP) (238422), puromycin (P8833), hygromycin (H3274), and blasticidin (15205) were purchased from Sigma. \u03b1 antibody (5324), anti\u2010GRP75 antibody (3593), anti\u2010HA antibody (3724), anti\u2010HIF\u20101\u03b1 antibody (36169), anti\u2010IRF3 antibody (4302), anti\u2010MCU antibody (14997), anti\u2010p\u20104E\u2010BP1 (Thr37/46) (2855), anti\u2010p62 antibody (88588), anti\u2010p70 S6 Kinase (2708), anti\u2010p\u2010Akt (Ser473) (4060), anti\u2010p\u2010eIF2\u03b1 (Ser51) antibody (3398), anti\u2010p\u2010Histone H2A.X (Ser139) antibody (9718), anti\u2010p\u2010IRF3 (Ser396) antibody (29047), anti\u2010PLK1 antibody (4513), anti\u2010p\u2010p70 S6 Kinase (Thr389) (9234), anti\u2010p\u2010S6 Ribosomal Protein (Ser235/236) (4858), anti\u2010p\u2010S6 Ribosomal Protein (Ser240/244) (5364), anti\u2010p\u2010STING (Ser366) (50907), anti\u2010p\u2010TBK1/NAK (Ser172) (5483), anti\u2010TBK1 antibody (51872), anti\u2010Skp2 antibody (2562), anti\u2010STING antibody (for WB and IP) (13647), anti\u2010STING antibody (for IF) (90947), anti\u2010ubiquitin antibody (3936), anti\u2010VDAC2 antibody (9412), anti\u2010rabbit IgG, HRP\u2010linked antibody (7074) and anti\u2010mouse IgG, HRP\u2010linked antibody (7076) were obtained from Cell Signaling Technology. Anti\u2010cyclin A antibody (sc\u2010751), anti\u2010GAPDH antibody (sc\u201047724), anti\u2010GST antibody (sc\u2010459), anti\u2010NQO1 antibody (sc\u201032793), anti\u2010p27\u2010antibody (sc\u20101641), anti\u2010PARP\u20101\u2010antibody (sc\u20108007), anti\u2010Ribosomal Protein S6 antibody (sc\u201074459) and anti\u2010vinculin antibody (sc\u201025336) were obtained from Santa Cruz Biotechnology. Polyclonal anti\u2010Flag antibody (F\u20107425), monoclonal anti\u2010Flag antibody , anti\u2010\u03b1\u2010tubulin antibody (T\u20105168), anti\u2010Flag agarose beads (A\u20102220), anti\u2010HA agarose beads (A\u20102095) and glutathione agarose beads (G4510) were obtained from Millipore Sigma. Anti\u2010ATP1A1 antibody (14418\u20101\u2010AP), anti\u2010COX IV antibody (11242\u20101\u2010AP), anti\u2010HIF\u20102\u03b1 antibody (26422\u20101\u2010AP), anti\u2010Histone H2A.X antibody (10856\u20101\u2010AP) and anti\u2010VDAC2 antibody (11663\u20101\u2010AP) were obtained from Proteintech. Anti\u2010Nrf2 antibody (ab62352) was obtained from Abcam. Invitrogen Alexa Fluor 488 goat anti\u2010rabbit IgG (H+L) (A11034), Invitrogen Alexa Fluor 594 goat anti\u2010mouse IgG (H+L) (A11032), and Invitrogen Alexa Fluor 594 Donkey anti\u2010Rabbit IgG (H+L) (A21207) were obtained from ThermoFisher Scientific.All antibodies were used at a 1:1000 dilution in TBST buffer with 5% bovine serum albumin or non\u2010fat milk for western blotting. Anti\u2010Akt antibody (4691), anti\u2010ATP2A2 antibody (9580), anti\u2010calnexin antibody (2679), anti\u2010cGAS antibody (83623), anti\u2010CHOP antibody (2895), anti\u2010c\u2010Myc antibody (18583), anti\u2010cyclin B1 antibody (4135), anti\u2010eIF2The renal cell carcinoma tumor specimens were obtained from the University of North Carolina at Chapel Hill Lineberger Comprehensive Cancer Center Tissue Procurement Facility reviewed and approved by the Office of Human Research Ethics as described previously.71\u22121 streptomycin unless otherwise stated.Human renal cell carcinoma cell lines 786\u2010o, A498, ACHN, Caki\u20101, RCC4, RCC10, RXF\u2010393, UMRC2, and UMRC6, human immortalized kidney cell lines HEK293, HEK293T, and HKC\u20108, human cervical adenocarcinoma cell line HeLa and human glioblastoma cell line U87MG were cultured in DMEM medium supplemented with 10% FBS. Human benign prostatic hyperplasia cell line BPH\u20101 cells were cultured in RPMI\u20101640 medium supplemented with 10% FBS. All cell culture media were supplemented with 100 units of penicillin and 100\u00a0mg\u00a0mL Packaging of lentiviral shRNA or cDNA expressing viruses, as well as subsequent infection of various cell lines were performed according to the protocols described previously. Following viral infection, cells were maintained in the presence of hygromycin (200\u00a0\u00b5g\u00a0mL\u22121) or puromycin (2\u00a0\u00b5g\u00a0mL\u22121), depending on the viral vector used to infect cells. Gene knockdown in shRNA\u2010tet\u2010on cells was achieved by refreshing medium containing 1\u00a0\u00b5g\u00a0ml\u22121 of doxycycline every 3 days.Cell transfection was performed using lipofectamine 3000 or polyethylenimine (PEI), as described previously.71 PacSTING into pCDNA3.0 vector using primer sets listed below. Flag\u2010STING\u2010HAQ\u2010H232R was constructed by PCR cloning HAQ\u2010H232R variant from THP\u20101 cell cDNA into pCDNA3.0 vector. Flag\u2010STING\u2010H232R was constructed by PCR cloning H232R variant from hORFeome V5.1 collection into pCDNA3.0 vector. Flag\u2010STING\u2010HAQ\u2010Y199C\u2010H232R was constructed by overlapping PCR to clone HAQ\u2010H232R variant into pCDNA3.0 vector using primers listed below. pBabe\u2010Flag\u2010STING\u2010WT, C91A and C88A/C91A were constructed by PCR cloning WT, C91A and C88A/C91A into pBabe\u2010hygro vector. pLenti\u2010Flag\u2010STING\u2010WT, C88A/C91A and R238A were constructed by cloning WT, R238A and C88A/C91A into pLenti\u2010hygro vector. CMV\u2010GST\u2010STING\u2010WT and S366A were constructed by (overlap) PCR cloning WT and S366A into CMV\u2010GST vector. Flag\u2010STING\u20101\u2010110, 1\u2013150, 42\u2013150, 82\u2013150, 111\u2013150, and 151\u2013379 were constructed by PCR cloning respective STING truncations into CMV\u2010GST vector using primer sets listed below. Flag\u2010cGAS was as described previously. His\u2010Ub was as described previously. HA\u2010VDAC2\u2010WT, C103A, C210A, C227A, C103A/C210A/C227A (3A), E84Q, Q101K, S128A, and L136F were constructed by (overlap) PCR cloning VDAC2 into pCDNA3 vector using primer sets listed below. pLenti\u2010HA\u2010VDAC2 and ATP2A2 were constructed by PCR cloning VDAC2 and ATP2A2 into pLenti\u2010GFP\u2010hygro vector using primer sets listed below. CMV\u2010GST\u2010GRP75 was constructed by PCR cloning GRP75 from hORFeome V5.1 collection into CMV\u2010GST vector. CMV\u2010GST\u2010ZDHHC7\u2010WT and C160S were constructed by (overlap) PCR cloning ZDHHC7 into CMV\u2010GST vector using primer sets listed below.Flag\u2010STING\u2010WT, C88A, C91A, C88A/C91A were constructed by (overlap) PCR cloning eviously.73 HisSTING\u2010BamHI\u2010F: GACACCGACTCTAGAGGATCCATGCCCCACTCCAGCCTGCASTING\u2010SalI\u2010Flag\u2010R: ATCCAGAGGTTGATTGTCGACTCACTTGTCGTCATCGTCTTTGTAGTC AGAGAAATCCGTGCGGAGAGSTING\u2010C88A\u2010F: TGGAGGACTGTGCGGGCCGCCCTGGGCTGCCCCCTCCGSTING\u2010C88A\u2010R: CGGAGGGGGCAGCCCAGGGCGGCCCGCACAGTCCTCCASTING\u2010C91A\u2010F: TGCGGGCCTGCCTGGGCGCCCCCCTCCGCCGTGGGGCCCSTING\u2010C91A\u2010R: GGGCCCCACGGCGGAGGGGGGCGCCCAGGCAGGCCCGCASTING\u2010C88A/C91A\u2010F: TGGAGGACTGTGCGGGCCGCCCTGGGCGCCCCCCTCCGCCGTGGGGCCCSTING\u2010C88A/C91A\u2010R: GGGCCCCACGGCGGAGGGGGGCGCCCAGGGCGGCCCGCACAGTCCTCCASTING\u2010P141A\u2010F: CAAGGGCCTGGCCGCAGCTGAGATCTCTGCAGTGSTING\u2010P141A\u2010R: CACTGCAGAGATCTCAGCTGCGGCCAGGCCCTTGSTING\u2010Y199C\u2010F: AGTGAGCCAGCGGCTGTGTATTCTCCTCCCATTGGSTING\u2010Y199C\u2010R: CCAATGGGAGGAGAATACACAGCCGCTGGCTCACTSTING\u2010R238A\u2010F: GCTGGCATCAAGGATGCGGTTTACAGCAACAGCATCTATGAGCSTING\u2010R238A\u2010R: GCTCATAGATGCTGTTGCTGTAAACCGCATCCTTGATGCCAGCSTING\u2010S366A\u2010F: AGCCTGAGCTCCTCATCGCTGGAATGGAAAAGCCCCTSTING\u2010S366A\u2010R: AGGGGCTTTTCCATTCCAGCGATGAGGAGCTCAGGCTSTING\u201042\u2010BamHI\u2010F: GCAT GGATCCATGCACACTCTCCGGTACCTGGTSTING\u201082\u2010BamHI\u2010F: GCAT GGATCCATGTGGAGGACTGTGCGGGCCTGSTING\u2010111\u2010BamHI\u2010F: GCAT GGATCCATGAATGCGGTCGGCCCGCCCTTSTING\u2010110\u2010SalI\u2010Flag\u2010R: GCATGTCGACTCACTTGTCGTCATCGTCTTTGTAGTCTGGGAGGGAGTAGTAGAAATSTING\u20101/150\u2010SalI\u2010Flag\u2010R: GCATGTCGACTCACTTGTCGTCATCGTCTTTGTAGTCTTTTTCACACACTGCAGAGASTING\u2010151\u2010BamHI\u2010F: GCATGGATCCATGGGGAATTTCAACGTGGCCCAVDAC2\u2010HA\u2010EcoRI\u2010F (for pCDNA3):GCATGAATTCATGTATCCATATGATGTTCCAGATTATGCTGCGACCCACGGACAGACTTGVDAC2\u2010HA\u2010XbaI\u2010F (for pLenti\u2010GFP hygro):GCATTCTAGAATGTATCCATATGATGTTCCAGATTATGCTGCGACCCACGGACAGACTTGVDAC2\u2010SalI\u2010R: GCAT GTCGAC TTAAGCCTCCAACTCCAGGGVDAC2\u2010C103A\u2010F: CAATTGAAGACCAGATTGCTCAAGGTTTGAAACTGAVDAC2\u2010C103A\u2010R: TCAGTTTCAAACCTTGAGCAATCTGGTCTTCAATTGVDAC2\u2010C210A\u2010F: CAATTTATCAGAAAGTTGCTGAAGATCTTGACACTTVDAC2\u2010C210A\u2010R: AAGTGTCAAGATCTTCAGCAACTTTCTGATAAATTGVDAC2\u2010C227A\u2010F: GGACATCAGGTACCAACGCCACTCGTTTTGGCATTGVDAC2\u2010C227A\u2010R: CAATGCCAAAACGAGTGGCGTTGGTACCTGATGTCCVDAC2\u2010E84Q\u2010F: GAGTATGGTCTGACTTTCACACAAAAGTGGAACACTGATAACACTCTGVDAC2\u2010E84Q\u2010R: CAGAGTGTTATCAGTGTTCCACTTTTGTGTGAAAGTCAGACCATACTCVDAC2\u2010Q101K\u2010F: CAGAAATCGCAATTGAAGACAAGATTTGTCAAGGTTTGAAACTGACVDAC2\u2010Q101K\u2010R: GTCAGTTTCAAACCTTGACAAATCTTGTCTTCAATTGCGATTTCTGVDAC2\u2010S128A\u2010F: GAAAAGTGGTAAAATCAAGTCTGCTTACAAGAGGGAGTGTATAAACCVDAC2\u2010S128A\u2010R: GGTTTATACACTCCCTCTTGTAAGCAGACTTGATTTTACCACTTTTCVDAC2\u2010L136F\u2010F: CAAGAGGGAGTGTATAAACTTTGGTTGTGATGTTGACTTTGATTTTGCVDAC2\u2010L136F\u2010R: GCAAAATCAAAGTCAACATCACAACCAAAGTTTATACACTCCCTCTTGATP2A2\u2010BglII\u2010HA\u2010F: GCATAGATCTATGTATCCATATGATGTTCCAGATTATGCTGAGAACGCGCACACCAAGACATP2A2\u2010Flag\u2010SalI\u2010R: GCATGTCGACTCACTTGTCGTCATCGTCTTTGTAGTCAGACCAGAACATATCGCTAAGRP75\u2010BamHI\u2010F: GCAT GGATCC ATGATAAGTGCCAGCCGAGCGRP75\u2010SalI\u2010R: GCAT GTCGAC TTACTGTTTTTCCTCCTTTTGZDHHC7\u2010BamHI\u2010F: GCATGGATCCATGCAGCCATCAGGACACAGZDHHC7\u2010SalI\u2010R: GCAT GTCGAC TCACACTGAGAACTCCGGGCZDHHC7\u2010sh3\u2010resist\u2010F: AGTGATTTTTCACCTCCCATTACAGTAATCCTGTTGATCTTZDHHC7\u2010sh3\u2010resist\u2010R: AAGATCAACAGGATTACTGTAATGGGAGGTGAAAAATCACTZDHHC7\u2010C160S\u2010F: GAAAATGGATCATCACTCCCCGTGGGTGAACAATTZDHHC7\u2010C160S\u2010R: AATTGTTCACCCACGGGGAGTGATGATCCATTTTCRT\u2010PCR primers are listed below:STING1\u2010F: CACTTGGATGCTTGCCCTCSTING1\u2010R: GCCACGTTGAAATTCCCTTTTTHMOX1\u2010F: AAGACTGCGTTCCTGCTCAACHMOX1\u2010R: AAAGCCCTACAGCAACTGTCGCYB5R3\u2010F: TCTACCTCTCGGCTCGAATTGCYB5R3\u2010R: CCTTGTCATCATCGCTGGAGATTXNIP\u2010qPCR\u2010F: ATATGGGTGTGTAGACTACTGGGTXNIP\u2010qPCR\u2010R: GACATCCACCAGATCCACTACTZDHHC3\u2010F: CCACTTCCGAAACATTGAGCGZDHHC3\u2010R: CCACAGCCGTCACGGATAAAZDHHC7\u2010F: CCCAAAGGAAACGCTACGAAAZDHHC7\u2010R: CGCGCTCGGGTTTAATACAGRNA18S\u2010F: TGCGGAAGGATCATTAACGGARNA18S\u2010R: AGTAGGAGAGGAGCGAGCGACCU6\u2010qPCR\u2010F: CTCGCTTCGGCAGCACAU6\u2010qPCR\u2010R: AACGCTTCACGAATTTGCGTPrimers used in generating plasmids used in this study are listed below:STING, cGAS, ZDHHC3, and ZDHHC7 were purchased from Sigma. Their target sequence is listed below:shScr: CCGGAACAGTCGCGTTTGCGACTGGCTCGAGCCAGTCGCAAACGCGACTGTTTTTTTTGshSTING\u20101: CCGGCCAACATTCGCTTCCTGGATACTCGAGTATCCAGGAAGCGAATGTTGGTTTTTTGshSTING\u20102: CCGGGCAGAGCTATTTCCTTCCACACTCGAGTGTGGAAGGAAATAGCTCTGCTTTTTTGshSTING\u20103: CCGGGTCCAGGACTTGACATCTTAACTCGAGTTAAGATGTCAAGTCCTGGACTTTTTTGshcGAS\u20101: CCGGCGTGAAGATTTCTGCACCTAACTCGAGTTAGGTGCAGAAATCTTCACGTTTTTTGshcGAS\u20102: CCGGATCTATTCTCTAGCAACTTAACTCGAGTTAAGTTGCTAGAGAATAGATTTTTTGshZDHHC3\u201010: CCGGGTATAGCATCATCAACGGAATCTCGAGATTCCGTTGATGATGCTATACTTTTTTGshZDHHC3\u201018: CCGGGCTTTGAAGAAGATTGGACAACTCGAGTTGTCCAATCTTCTTCAAAGCTTTTTTGshZDHHC3\u201079: CCGGCCAGAAGTACTTCGTCCTGTTCTCGAGAACAGGACGAAGTACTTCTGGTTTTTTGshZDHHC7\u20103: CCGGGATAACTGTAATCCTGTTGATCTCGAGATCAACAGGATTACAGTTATCTTTTTTGshZDHHC7\u201034: CCGGACTGCCCGTGGGTGAACAATTCTCGAGAATTGTTCACCCACGGGCAGTTTTTTTGshRNA plasmids to deplete endogenous shMCU: CCGGGCAAGGAGTTTCTTTCTCTTTCTCGAGAAAGAGAAAGAAACTCCTTGCTTTTTG\u03b1: CCGGCCAGTTATGATTGTGAAGTTACTCGAGTAACTTCACAATCATAACTGGTTTTTTGshHIF\u20101\u03b1: CCGGGGAGACGGAGGTGTTCTATTTCAAGAGAATAGAACACCTCCGTCTCCTTTTTTGshHIF\u20102shGRP75\u201051:CCGGGCACATTGTGAAGGAGTTCAACTCGAGTTGAACTCCTTCACAATGTGCTTTTTTGshGRP75\u201052:Other shRNA plasmids were constructed by inserting synthesized shRNAs into pLKO\u2010puro or pLKO\u2010hydro vectors. Their target sequence is listed below:shVDAC2: CCGGAAGGATGATCTCAACAAGAGCCTCGAGGCTCTTGTTGAGATCATCCTTTTTTTTGTet\u2010inducible shRNA plasmids were constructed by inserting synthesized shRNAs into Tet\u2010pLKO\u2010neo vector. Target sequence of STING was the same as that of shSTING\u20102 and \u20103. Target sequence of shVDAC2 is listed below:sgSTING\u20101B: GCTGGGACTGCTGTTAAACGsgSTING\u20106: CATTACAACAACCTGCTACGsgcGAS\u20101: CACGTGCTCATAGTAGCTCCsgcGAS\u20102: GGCCGCCCGTCCGCGCAACTsgIRF3\u20101: CGCTCACTGCCCAGTATGTGsgIRF3\u20104: GGCACCAACAGCCGCTTCAGsgMCU: GTGTTTTCTAGGTACACCAGsgRNA plasmids were constructed by inserting synthesized sgRNAs into lentiCRISPRv2\u2010puro vector. Their target sequence is listed below:STING\u2010S366A\u2010KI\u2010sgF: CACCGGCTTTTCCATTCCACTGATGSTING\u2010S366A\u2010KI\u2010sgR: AAACCATCAGTGGAATGGAAAAGCCSTING\u2010S366A\u2010KI\u2010ssODNAGGGCAGCTTGAAGACCTCAGCGGTGCCCAGTACCTCCACGATGTCCCAAGAGCCTGAGCTATTAATCGCTGGAATGGAAAAGCCCCTCCCTCTCCGCACGGATTTCTCTTGAGACCCAGGGTCACCAGGCCAGAGCCTCCSTING\u2010S366A knockin experiment was performed using STING sgRNAs and ssoDNA as listed below:STING\u2010S366A\u2010KI\u2010PCR\u2010F: GAGTGGGAATGGGTAAGATCCTCSTING\u2010S366A\u2010KI\u2010PCR\u2010R: GACGCATCTTAAGATGTCAAGTCCKnockin clones were screened by PCR using primers listed below to search for clones loss of SacI but gain of AseI site after knockin. Equal amounts of whole cell lysates were resolved by SDS\u2010PAGE and immunoblotted with indicated antibodies. For immunoprecipitations analysis, unless specified, 1\u00a0mg lysates were incubated with the indicated antibody (1\u20132\u00a0\u00b5g) for 3\u20134\u00a0h at 4\u00a0\u00b0C followed by 1\u00a0h incubation with 10\u00a0\u00b5L Protein A/G XPure Agarose Resin . Or 1\u00a0mg lysates containing tagged molecules were incubated with agarose beads coupled antibodies for the specific tag for 3\u20134\u00a0h at 4\u00a0\u00b0C. For endogenous IPs, incubation of cell lysates with antibodies was extended to overnight. The recovered immuno\u2010complexes were washed five times with NETN buffer before being resolved by SDS\u2010PAGE and immunoblotted with indicated antibodies. For VDAC2 oligomerization detection, cells were incubated with 250\u00a0\u00b5M ethylene glycol bis (succinimidyl succinate) , lyzed in Triton X\u2010100 buffer and nixed with 4 \u00d7 lithium dodecyl sulfate sample buffer . Load protein without boiling in SDS\u2010free PAGE gel and immunoblotted with indicated antibodies.Cells were lysed in EBC buffer or Triton X\u2010100 buffer supplemented with protease inhibitor cocktail and phosphatase inhibitor cocktail . The protein concentrations of whole cell lysates were measured by NanoDrop OneC using the Bio\u2010Rad protein assay reagent as described previously.71 Equ 1\u2010day post\u2010transfection, cells were selected with 1\u00a0\u00b5g\u00a0ml\u22121 puromycin for 3 days. Surviving cells were counted and each single cell was seeded into 96\u2010well plates. Each single clone grown up in 96\u2010well plates was amplified and one copy was used for genomic DNA extraction, followed by PCR and AseI/SacI digestion to screen for potential knockin clones. SacI negative but AseI positive clones are selected and sequenced to verify the knockin at the DNA level.Parental A498 cells were split into 24\u2010well plates and transfected with sgRNA against endogenous STING together with STING\u2010S366A\u2010ssoDNA following protocols as described.60 1\u2010dm urea to 100\u00a0\u00b5g\u00a0\u00b5l\u22121 and subjected to FASP trypsin digestion protocol. Briefly, proteins were reduced using 50\u00a0mM DTT for 15\u00a0min at 65\u00a0\u00b0C. Proteins were then transferred to a 30\u00a0000 MWCO spin filter , diluted with 200\u00a0\u00b5l of 8\u00a0m urea, and centrifuged at 10\u00a0000 \u00d7 g for 30\u00a0min at RT. The proteins were washed twice using 200\u00a0\u00b5l of 8\u00a0m urea\u00a0followed by 30 minutes of centrifugation at 10\u00a0000 \u00d7 g between each wash.\u00a0Proteins were alkylated using 100\u00a0\u00b5l of 15\u00a0mM 2\u2010chloroacetamide prepared in 8\u00a0m urea for 20\u00a0min in the dark at RT. The spin filter was then centrifuged at 10\u00a0000\u00a0\u00d7\u00a0g for 20\u00a0min at RT followed by two washes with 200\u00a0\u00b5l of 8\u00a0M urea with 20\u00a0min of centrifugation at 10000\u00a0\u00d7\u00a0g in between each wash. Buffer exchange into 50\u00a0mM ammonium bicarbonate (ABC) pH 8.0 was performed with two washes of 200\u00a0\u00b5l of 50\u00a0mM ABC with centrifugation at 10\u00a0000\u00a0\u00d7\u00a0g for 15\u00a0min at RT between each wash. 100\u00a0\u00b5l of 50\u00a0mM ABC was added to the spin filter along with 2.5\u00a0\u00b5g of trypsin (Promega V511C). Proteins were trypsinized overnight at 37\u00b0C for 18\u00a0h. Following trypsinization, peptides were recovered in a new receiver tube by centrifugation at 10\u00a0000\u00a0\u00d7\u00a0g for 15\u00a0min. Peptides were eluted twice\u00a0using 50\u00a0\u00b5l of 0.5% TFA in water at 10\u00a0000\u00a0\u00d7\u00a0g for 10\u00a0min.\u00a0Samples were then concentrated to 100\u00a0\u00b5l using a\u00a0speedvac followed by C18 desalting . Samples were then concentrated using a speedvac and resolubilized in 100\u00a0\u00b5l of LC\u2010Optima\u00a0MS\u2010grade water (Thermo). Ethyl acetate extraction followed by speedvac was performed to remove residual detergents.\u00a0QFP assay was performed for peptide quantification.A498 cells stably expressing either FLAG\u2010EV or FLAG\u2010STING were lysed in EBC lysis buffer FLAG immunoprecipitants were washed with NETN buffer. Proteins were diluted in 8\u00a0 Data was searched using MaxQuant (version 1.6.6.7), and all statistical analyses were done in Perseus (version 1.6.3.4).Detailed liquid chromatography\u2010mass spectrometry/mass spectrometry\u00a0methods and data filtering methods were described previously.76 DatRNA extraction was performed with an RNA miniprep super kit and QIAshredder . The final elution step was done with 50\u00a0\u00b5L of RNAse\u2010free water. The relative enrichment of mRNA was quantified with the NanoDrop OneC (ThermoFisher Scientific). At least two biological replicates were performed for RNA extraction. Reverse transcription was performed with iScript cDNA synthesis kit . Quantitative real\u2010time PCR was performed with iTaq universal SYBR green supermix using a QuantStudio\u202f6 Flex Real\u2010Time PCR Systems (ThermoFisher Scientific). Each mRNA level was normalized to RNA18S or U6 snRNA. The comparative Ct method was used to calculate fold change in expression. Statistical significance was determined by one\u2010way ANOVA tests.2) for 3\u00a0h. After thorough mixing, absorbance at 450\u00a0nm was measured using the BioTek Cytation 5 Cell Imaging reader. Combination index (CI) of co\u2010treatment of 2\u2010BP and sorafenib was calculated by CompuSyn software .Indicated number of cells were seeded in each well of 96\u2010well plates for cell viability assay to monitor cell viability at indicated time periods using Cell Counting Kit 8 according to manufacturer's instruction. Briefly, at indicated time points post\u2010cell seeding, 10\u00a0\u00b5L CCK8 solution was added into each well and incubated in the culture incubator (37\u00a0\u00b0C with 5% CO2 for 7\u201315 days (as indicated in figure legends) until formation of visible colonies. Colonies were washed with 1x PBS, fixed with methanol for 30\u00a0min, and stained with 0.5%\u00a0crystal violet for 30\u00a0min. Colonies were then washed with distilled water and air\u2010dried. Colony numbers were manually counted. Three independent experiments were performed to generate the error bars.Indicated cells were seeded into 6\u2010well or 24\u2010well plates (500 cells/well) and cultured in 37\u00a0\u00b0C incubator with 5% CO Briefly, the assays were preformed using 6\u2010well plates where the solid medium consists of two layers. The bottom layer contains 0.8% noble agar, and the top layer contains 0.4% agar suspended with 3\u00a0\u00d7 104 or indicated number of cells. 500\u00a0\u00b5L complete DMEM medium with 10% FBS was added every 4 days. About 4\u20136 weeks later the cells were stained with iodonitrotetrazolium chloride (1\u00a0mg\u00a0mL\u22121) (Sigma I10406) overnight for colony visualization, imaging, and counting. Three independent experiments were performed to generate the error bar.The anchorage\u2010independent cell growth assays were performed as described previously.53 Bri Briefly, for mouse xenograft growth experiments, A498 cells were infected with viruses expressing shScr or shSTING2/3. Two days later, 1\u00a0\u00d7 106 A498 cells in PBS were injected into the flank of indicated female nude mice . Tumor size was measured every two days with a digital caliper, and the tumor volume was determined with the formula: L \u00d7 W2 \u00d7 0.5, where L is the longest diameter and W is the shortest diameter. After 66 days, mice were sacrificed, and tumors were dissected and weighed. For inducible STING shRNA, 1\u00a0\u00d7 107 A498 cells in DMEM were injected into flank of male nude mice . When tumors became visible after 15 days, 2\u00a0mg\u00a0ml\u22121 doxycycline was added to drinking water containing 2% sucrose. Water was changed every three days and protected from light. Mice were sacrificed 39 days after injection. For combination therapy, 1\u00a0\u00d7 107 A498 cells in DMEM were injected into flank of male nude mice . After 25 days, when tumors reached a volume of \u2248150 mm3, mice were randomly divided into four groups. 2\u2010BP was dissolved in 10% DMSO+40% PEG300+5% Tween 80+ 45% saline. Sorafenib was dissolved in 3% DMSO+97% sulfobutylether\u2010\u03b2\u2010Cyclodextrin solution . Mice were daily treated with 40\u00a0mg\u00a0kg\u22121 2\u2010BP given by intraperitoneal injection or 40\u00a0mg\u00a0kg\u22121 sorafenib given by oral gavage or both drugs or their solvents. Tumor size was measured regularly, and the tumor volume was determined with the formula: L x W2 x 0.5, Mice were sacrificed after 13 days of treatment.All mouse work has been reviewed and approved by UNC Institutional Animal Care and Use Committee under IACUC#19\u2010031 which has been continued as #22\u2010056. Mouse xenograft assays were performed as described previously.71 Bri Reads overlapping blacklisted regions of the genome were then removed. Transcript abundance was then estimated using Salmon, and differentially expressed genes were detected using DESeq2 with the criteria of adjusted p\u2010values (adjP) <0.05. The ClusterProfiler R package (v3.14.3) was employed to analyze the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) for functional pathway annotation. Enrichment analysis for GO terms and KEGG pathways utilize enrichGO and enrichKEGG functions and visualizes the result with bubble plots. RNA\u2010seq data are deposited to GEO under accession number GSE190816.Total RNA from triplicate samples of A498 cells infected with shScr or shSTING was extracted with RNeasy mini kit . Library preparation and sequencing were performed by GENEWIZ as paired\u2010end 150\u2010bp reads. Reads were then filtered for adaptor contamination using cutadapt and filtered such that at least 90% of bases of each read had a quality score > 30. Reads were aligned to the reference genome (hg19) using STAR version 2.5.2b retaining only primary alignments.77 Rea4, 1\u00a0\u00b5M Alexa 647\u2010azide (Life Technologies), and 100\u00a0mM ascorbic acid (fresh) for 30\u00a0min. at room temperature in the dark. Finally, cells were resuspended in 1% BSA\u2010PBS with 1\u00a0\u00b5g\u00a0mL\u22121 DAPI (Life Technologies) and 100\u00a0\u00b5g\u00a0mL\u22121 RNAse A (Sigma Aldrich) and incubated overnight at 4\u00a0\u00b0C in the dark. Data were collected on an Attune NxT flow cytometer (Thermo Fisher Scientific) and analyzed using FCS Express 7 software (De Novo Software).To measure cell cycle phase distribution, cells were treated with 10\u00a0\u00b5M EdU for 30\u00a0min. before harvesting by trypsinization. Cells were washed with PBS and then fixed in 4% paraformaldehyde in PBS for 15\u00a0min. at room temperature. 1% BSA\u2010PBS was added, mixed, and cells were centrifuged. Fixed cells were permeabilized with 0.5% triton X\u2010100 in 1% BSA\u2010PBS at room temperature for 15\u00a0min. and centrifuged. Cells were then processed for EdU detection as follows: samples were incubated in PBS with 1\u00a0mM CuSOTo measure general ROS level, trypsinized cells were incubated with 10\u00a0\u00b5M CM\u2010H2DCFDA in PBS at 37\u00a0\u00b0C for 30\u00a0min. To measure mitochondrial ROS level, trypsinized cells were incubated with 1\u00a0\u00b5M MitoSOX Red mitochondrial superoxide indicator in PBS at 37\u00a0\u00b0C for 20\u00a0min. To measure mitochondrial calcium level, trypsinized (without EDTA) cells were incubated with 5\u00a0\u00b5M Rhod\u20102, AM in PBS solution containing 10% FBS and 10\u00a0mM glucose at 37\u00a0\u00b0C for 1\u00a0h. To measure mitochondrial membrane potential, cells were incubated with 2\u00a0\u00b5M JC\u20101 in PBS at 37\u00a0\u00b0C for 15\u00a0min. Cells were washed with PBS twice and analyzed with BD FACScanto II or BD FACSfortessa (BD Biosciences) with the BD Diva software (BD Biosciences). A minimum of 10\u00a0000 events were acquired for each sample. Data were analyzed using Flowjo 7.6.1 (Tree Star).Cells plated onto glass coverslips were fixed with 4% paraformaldehyde in PBS for 20\u00a0min at room temperature and permeabilized with 0.2% Triton X\u2010100 for 20\u00a0min at room temperature. Cells were incubated with blocking buffer for 1\u00a0h, incubated with primary antibodies at 4\u00a0\u00b0C overnight, incubated with secondary antibodies and/or mitochondria dye MitoView Green at room temperature for 1\u00a0h and mounted with ProLong Gold antifade reagent . Fluorescent signals were observed with an Olympus 1 \u00d7 51 inverted microscope at \u00d7 20 magnification, or with an Olympus FV1000 confocal microscope at 60\u00d7 magnification.\u22121. Added 800\u00a0\u00b5l reagent C and invert tubes several times to mix. Centrifuged tubes at 700 \u00d7 g for 10\u00a0min and collect supernatant as a mixture containing mitochondria, lysosome, and peroxisome. Centrifuged supernatant at 3000 \u00d7 g for 15\u00a0min and this pellet contains isolated mitochondria. Centrifuged supernatant at 12\u00a0000 \u00d7 g for 15\u00a0min and transferred new supernatant containing cytosolic fraction to a new tube. The pellet was a mixture of lysosome, peroxisome, and some mitochondria. Resuspended mitochondria pellet in 500\u00a0\u00b5l reagent C and centrifuged at 12\u00a0000 \u00d7 g for 5\u00a0min to obtain pure mitochondrial fraction. Samples were kept at 4\u00a0\u00b0C throughout the procedure.Cytosolic and mitochondrial fractions of cells were isolated with Mitochondria isolation kit for cultured cells according to manufacturer's instruction. Briefly, add 800\u00a0\u00b5l reagent A to cell pellets and incubate for 2\u00a0min. Added 10\u00a0\u00b5l reagent B and vortex 5 s\u00a0min Image analysis was performed using Adobe Illustrator.Electron microscopic analysis of indicated A498 cells was performed as detailed below. Sample prep: indicated A498 cell culture pellets were embedded in 2% agarose, secondary fixation with osmium tetroxide, dehydration , followed by embedding in EPON.\u00a0Sectioning: \u224880\u00a0nm thin sections on Cu grids, stained with UA and LC. Imaging: FEI Tecnai 12 at 120\u00a0kV, Gatan Rio16 CMOS camera. Cells were randomly selected for imaging. Images at 4400\u00d7 magnification were used for counting total and abnormal mitochondria. All clearly identified mitochondria in the images were analyzed. Images at 26\u00a0000\u00d7 magnification were randomly selected for mitochondria\u2010ER contact (MERC) length measurement. All clearly identified mitochondria membranes and associated ER membranes in the images were evaluated. Only MERC sites with a distance no more than 30\u00a0nm were regarded as MERC enabling calcium transfer from ER to mitochondria and their length was measured according to.80 Ima4 cells were seeded into XF24 cell culture microplate before the assay. Added 1\u00a0mL of Seahorse XF24 calibrant solution (pH 7.4) to each well of XF24 microplate and replaced green sensor cartridge on the top. Incubated entire cartridge in a non\u2010CO2 incubator at 37\u00b0C overnight. On the day following cell seeding, media was changed to phenol red\u2010free, Seahorse XF DMEM medium, pH 7.4 supplemented with 25\u00a0mM glucose and 1\u00a0mM sodium pyruvate for OCR or 2\u00a0mM glutamine and 1\u00a0mM sodium pyruvate for ECAR. Cells were equilibrated within 1\u00a0h in 37\u00a0\u00b0C non\u2010CO2 incubator. Loaded the sensor cartridge for the calibration process and replaced calibration plate with cell plate after that. The basal mitochondrial respiration or extracellular acidification rate was first measured by recording extracellular oxygen concentration. Then the OCR or ECAR trace was recorded in response to sequential addition of indicated compounds in the Seahorse XF Cell Mito Stress Test Kit or Glycolysis Stress Test Kit . Concentration of compounds for OCR was: 1\u00a0\u00b5M of oligomycin A, 1\u00a0\u00b5M of FCCP, and 1\u00a0\u00b5M of rotenone/antimycin A. Concentration of compounds for ECAR was: 10\u00a0mM of glucose, 1\u00a0\u00b5M of oligomycin, and 50\u00a0mM of 2\u2010DG. 3 to 5 technical replicates were utilized per sample to calculate OCR or ECAR.The OCR and ECAR were measured by an XFe24 extracellular flux analyzer (Agilent Technologies), according to the manufacturer's instructions. Briefly, 5 \u00d7 10 Briefly, cells were lysed in lysis buffer (LB) supplemented with protease inhibitor, PMSF and 50\u00a0mM N\u2010ethylmaleimide (NEM) . Equal amounts of lysates were incubated with the STING antibody overnight at 4\u00a0\u00b0C followed by 1\u00a0h incubation with 60\u00a0\u00b5L Protein A/G XPure Agarose Resin, or incubated with Flag agarose beads for 3\u20134\u00a0h at 4\u00a0\u00b0C. Beads were resuspended in LB+10\u00a0mM NEM and split into triplicates. 1/3 of beads were used as \u2010HAM control and 2/3 were used as +HAM treatment. Beads were resuspended with LB (pH 7.2)+0.1% SDS quickly, washed with LB (pH\u00a0= 7.2) three times, resuspended in LB (pH 7.2) with or without 1\u00a0m hydroxylamine (HAM) and incubated at room temperature for 1\u00a0h. Then, beads were washed with LB (pH 6.2) once and resuspended in LB (pH\u00a0= 6.2)+2\u00a0\u00b5M Biotin\u2010BMCC at 4\u00a0\u00b0C for 1\u00a0h. Beads were then washed with LB (pH\u00a0= 6.2) once and LB (pH 7.5) for three times. Beads were boiled and resolved by SDS\u2010PAGE and immunoblotted with streptavidin\u2010HRP antibody to detect biotin\u2010labeled palmitoylated STING.Protein palmitoylation detection was performed by immunoprecipitation and acyl\u2010biotin exchange as described previously.81 Brip \u2264 0.05 was considered statistically significant. The results were shown as means \u00b1 SD from at least two or three independent experiments as indicated in figure legends. Differences between control and experimental conditions were evaluated by One\u2010way ANOVA.Statistical analyses were performed using the Graphpad Prism 8 Software. The authors declare no conflict of interest.Conceptualization: ZZ, PL; Methodology: ZZ, JGC, BM, XT, HY, GD, PL; Investigation: ZZ, HD, XZ, EWC, JZ, LM, YW, XT, TS, JS; Visualization: ZZ, YW, JZ; Funding acquisition: JGC, DH, GD, PL; Project administration: JLS, EH; Supervision: JGC, BM, GD, PL; Writing \u2013 original draft: ZZ, PL; Writing \u2013 review & editing: HY, JGC, BM, GD.Supporting InformationClick here for additional data file."} +{"text": "The crystal structure of the title salt is held by N\u2014H\u22efBr charge-assisted hydrogen bonds and dipole\u2013dipole inter\u00adactions. In comparison with reference structures, elongation of C\u2014S bonds is observed. 12H13N2S+\u00b7Br\u2212, which crystallizes in the monoclinic P21/c centrosymmetric space group. The asymmetric unit contains one 2-[meth\u00adyl]iso\u00adthio\u00aduronium cation and one bromide anion. The methyl\u00adene carbon lies in plane of the naphthalene core. In comparison with reference structures, elongation of C\u2014S bonds as well as tilting of the iso\u00adthio\u00aduronium group is observed. Given the ionic nature of the compound, the structure is held by charge-assisted N\u2014H\u22efBr hydrogen bonds, with a noteworthy contribution of dipole\u2013dipole inter\u00adactions, which form bilayers in the structure. The bilayers are held by the weak London forces.Herein we report the crystal structure of 2-[meth\u00adyl]iso\u00adthio\u00aduronium bromide, C Since the relevant torsion angle of the title compound is \u221268.1\u2005(3)\u00b0, only the non-linear 2-benzyl\u00adiso\u00adthio\u00aduronia, will be used as a reference group [CCDC codes EBIFOK meth\u00adyl]iso\u00adthio\u00aduronia structures in the the CSD; therefore, we decided to compare the title compound with 2-benzyl\u00adiso\u00adthio\u00aduronia structures. The published structures of 2-benzyl\u00adiso\u00adthio\u00aduronia can be divided according to the CThe C11\u2014S1 bond is almost perpendicular to the naphthalene core, with C1\u2014C2\u2014C11\u2014S1 and C3\u2014C2\u2014C11\u2014S1 torsion angle values of 91.8\u2005(3) and \u221288.0\u2005(3)\u00b0, respectively. Among the reference group, such a conformation is unusual, with average values being 61.45 and 120.53\u00b0. The iso\u00adthio\u00aduronium group is significantly tilted, with C11\u2014S1\u2014C12\u2014N1 and C11\u2014S1\u2014C12\u2014N2 torsion angles of \u221268.5\u2005(3) and 110.7\u2005(3)\u00b0, respectively. Such a tilt is not observed among the reference group, where the average values are 20.46 and 160.94\u00b0. The tilting of the group can be explained by the steric demands of the 2-naphthyl\u00admethyl unit on the packing.n1\u22efBr1\u22efH2n1iii\u2014N1iii, and N2\u2014H1n2\u22efBr1ii\u22efH2n2ii\u2014N2ii classified as n1\u22efBr1\u22efH1n2iv\u2014N2iv\u2014C12iv\u2014N1iv, N1\u2014H2n1\u22efBr1i\u22efH1n2v\u2014N2v\u2014C12v\u2014N1v, and N1\u2014H2n1\u22efBr1i\u22efH2n2i\u2014N2i\u2014C12i\u2014N1i classified as x, y\u00a0\u2212\u00a01, z; (ii) x, \u2212y\u00a0+\u00a0z\u00a0\u2212\u00a0x, y\u00a0+\u00a01, z; (iv) x, \u2212y\u00a0+\u00a0z\u00a0+\u00a0x, \u2212y\u00a0+\u00a0z\u00a0+\u00a0vi iso\u00adthio\u00aduronium bromide.The 2-[meth\u00adyl]iso\u00adthio\u00aduronium bromide (20\u2005mg) was dissolved in 10\u2005ml of methanol and left to slowly evaporate at room temperature. After 5\u2005d, colorless platelets were collected.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314620015114/sj4217sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314620015114/sj4217Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314620015114/sj4217Isup3.molSupporting information file. DOI: 2044275CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The coordination sphere around each Cd2+ ion is completed by three N atoms of a tridentate 8-[2-(di\u00admethyl\u00adamino)\u00adethyl\u00adamino]quinoline ligand and another chloride ion to form a distorted fac-CdN3Cl3 octa\u00adhedron. The ethanol mol\u00adecule is both an acceptor of an N\u2014H\u22efO and a donor of an O\u2014H\u22efCl hydrogen bonds to its adjacent complex unit. In the crystal, weak aromatic \u03c0\u2013\u03c0 stacking is observed.The title solvated bimetallic complex, [Cd Similar perpendicular arrangement of the pendant Cl\u2212 ions is observed in the Cl\u2013(Cd2Cl2)\u2013Cl fragments of other complexes with different ligands \u00b0 (N3\u2014Cd1\u2014N2) to 105.73\u2005(3)\u00b0 (Cl1\u2014Cd1\u2013Cl2) for one Cd2+ ion and 71.04\u2005(9) \u00b0 (N6\u2014Cd2\u2014N5) to 102.09\u2005(7)\u00b0 (N5\u2014Cd2\u2014Cl2) for the other. The corres\u00adponding angles for the hydrate structure are in the range 69.48\u2005(5) to 101.08\u2005(4)\u00b0. The N\u2014C\u2014C\u2014N torsion angles in the ethane di\u00adamine are almost the same for both independent ligands [N1\u2014C3\u2014C4\u2014N2 = 63.0\u2005(4)\u00b0 and N4\u2014C16\u2014C17\u2014N5 = 63.3\u2005(5)\u00b0] in I.Both CdR(6)22 loop. In the extended structure, the quinoline ring systems of neighbouring complex units are involved in weak aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions. The groups involved are related by inversion symmetry with a c(i)\u22efc(i)\u2032 separation of 3.93\u2005(1)\u2005\u00c5 [c(i) = the midpoint of the C9\u2014C10 bond of the C5\u2013C13/N3 ring system]. If a second longer inversion-related contact c(ii)\u22efc(ii)\u2032 of 4.56\u2005(1)\u2005\u00c5 [c(ii) = midpoint of the C22\u2014C23 bond of the C18\u2013C26/N6 ring system] is considered to be a significant inter\u00adaction, infinite chains running parallel to [101] result \u00adethyl\u00adamino]\u00adquinoline ligand and cadmium dichloride were mixed in dry ethanol solvent at room temperature under a positive nitro\u00adgen pressure and the mixture was stirred at room temperature for several hours. The solution was then warmed to dissolve the material and the product was recrystallized on cooling to produce colourless crystals of Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621001504/hb4375sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314621001504/hb4375Isup2.hklStructure factors: contains datablock(s) I. DOI: 2062006CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of a resveratrol\u2013oxime ester is reported. 22H25NO6, the C=C double bond linking the benzene rings adopts an E configuration and the dihedral angle between the rings is 47.1\u2005(2)\u00b0. The oxime unit contains a C=N double bond, which also has an E configuration. In the crystal, pairs of C\u2014H\u22efN hydrogen bonds generate inversion dimers and weak C\u2014H\u22efO inter\u00adactions link the dimers into chains propagating along the b-axis direction.In the title compound, C The dihedral angle formed by benzene rings is 47.1\u2005(2)\u00b0. The C17=N1 imine double bond in the oxime unit also adopts an E configuration, which is defined by a torsion angle of 178.3\u2005(1)\u00b0 for C4\u2014C17\u2014N1\u2014O3. There are three meth\u00adoxy groups attached to carbon atoms C1, C5 and C13 in the benzene rings: those at the meta positions are essentially co-planar with their attached benzene rings [C6\u2014C1\u2014O1\u2014C7 = \u22120.2\u2005(2)\u00b0 and C6\u2014C5\u2014O6\u2014C22 = 3.9\u2005(2)\u00b0] whereas the meth\u00adoxy group at the para position (C13) is slightly twisted from the corresponding ring plane [C12\u2014C13\u2014O2\u2014C16 = 8.9\u2005(2)\u00b0]. In the crystal, pairs of C22\u2014H22\u22efN1 hydrogen bonds generate inversion dimers benz\u00adaldehyde I. DOI: 10.1107/S2414314621009500/hb4391Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314621009500/hb4391Isup3.cmlSupporting information file. DOI: 2109339CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Using the equation-of-motion coupled-cluster singles and doubles method and Fermi\u2019s golden rule, we quantitatively reproduced all rate constants relevant to the emission mechanism; prompt and delayed fluorescence, internal conversion (IC), intersystem crossing, and reverse intersystem crossing (RISC). In addition, the photoluminescence quantum yield and its prompt and delayed contributions were quantified by calculating the population kinetics of excited states and the transient photoluminescence decay curve. The calculations also revealed that TADF occurred via a stepwise process of 1)\u00a0thermally activated IC from the electronically excited lowest triplet state T1 to the second-lowest triplet state T2, 2)\u00a0RISC from T2 to the lowest excited singlet state S1, and 3)\u00a0fluorescence from S1.Molecules that exhibit multiple resonance (MR) type thermally activated delayed fluorescence (TADF) are highly efficient electroluminescent materials with narrow emission spectra. Despite their importance in various applications, the emission mechanism is still controversial. Here, a comprehensive understanding of the mechanism for a representative MR-TADF molecule are highly efficient electroluminescent materials, but their emission mechanism is still not fully understood. Here, the emission mechanism of a representative MR-TADF molecule is studied through quantum chemistry calculations, with two consecutive decay processes unveiled. An important factor for TADF efficiency is the energy difference between the lowest triplet state (T1) and the lowest excited singlet state (S1), \u2206E(T1\u2009\u2192\u2009S1); a small \u2206E(T1\u2009\u2192\u2009S1) (<200\u2009meV) is required to induce T1\u2009\u2192\u2009S1 reverse intersystem crossing (RISC). Another important factor is a large transition-dipole moment between S1 and ground-state S0 to accelerate the rate of S1\u2009\u2192\u2009S0 fluorescence. Experimentally, efficient RISC and high photoluminescence quantum yields (PLQYs) have been simultaneously realized by combining suitable donor and acceptor units that control the spatial overlap between the highest-occupied molecular orbitals (HOMOs) and the lowest-unoccupied molecular orbitals (LUMOs)9. It is important to note that the fluorescence spectra were broadened because of the charge-transfer character of S1, which was a significant drawback for OLED applications in displays.Since highly efficient thermally activated delayed fluorescence (TADF) was used in organic light-emitting diodes (OLEDs)10. Using a triphenylboron core possessing two nitrogen atoms . However, \u2206E(T1\u2009\u2192\u2009S1) and \u2206E(T1\u2009\u2192\u2009T2) were large, 0.59\u2009eV and 0.61\u2009eV, respectively, which could not explain efficient RISC from T1 to S1. Gao et al. examined the density-functional dependence of \u2206E(T1\u2009\u2192\u2009S1) within the framework of TD-DFT41. The MPWK1CIS functional reproduced the experimental \u2206E(T1\u2009\u2192\u2009S1)41, as reviewed by Suresh et al.21. However, the RISC rate constant (kRISC) was not calculated and the TADF mechanism was unclear. Also, they considered TADF in terms of direct (one-step) T1\u2009\u2192\u2009S1 RISC, which differed from the work of Northey et al40. Pershin et al. reported that TD-DFT methods overestimated \u2206E(T1\u2009\u2192\u2009S1) for MR molecules, and that the spin-component-scaling second-order approximate coupled-cluster (SCS-CC2) method outperformed TD-DFT for predicting \u2206E(T1\u2009\u2192\u2009S1)43. The partial inclusion of double excitations within the SCS-CC2 method was responsible for the improved accuracy in predicting \u2206E(T1\u2009\u2192\u2009S1). The SCS-CC2 calculation of DABNA-1-based molecules revealed the relationship between its molecular structure and electronic properties, \u2206E(T1\u2009\u2192\u2009S1), and the fluorescence rate constant. However, the TADF mechanism was still unclear because the kRISC was not calculated. Lin et al. calculated rate constants for fluorescence (kF), ISC (kISC), kRISC, and internal conversion (IC) (kIC) for DABNA-144. However, the calculated kRISC was 6.7\u2009\u00d7\u2009102\u2009s\u22121, which was much less than the experimental value of 1.0\u2009\u00d7\u2009104\u2009s\u2212110. The calculated kISC of 1.4\u2009\u00d7\u2009104\u2009s\u22121 was two orders of magnitude less than the experimental value of 4.5\u2009\u00d7\u2009106\u2009s\u2212110. In addition, the calculated nonradiative decay, kIC(S1\u2009\u2192\u2009S0), was greater than the kF(S1\u2009\u2192\u2009S0), suggesting that the PLQY of DABNA-1 was less than 50%, in contrast with the 88% experimental value10. All these previous studies44 only partially explained the photophysical properties of MR-TADF mainly because crucial RISC was not elucidated.Several theoretical studies have attempted to reveal the TADF mechanism in MR molecules. Northey et al. investigated an intersystem-crossing (ISC) mechanism in DABNA-1 using quantum dynamics and time-dependent density-functional theory (TD-DFT) at the PBE0/6-31\u2009G(d) levelE(T1\u2009\u2192\u2009S1) (and spin\u2013orbit coupling (SOC) in some cases) and oscillator strength have been considered to understand the TADF mechanism and to design TADF molecules. However, these are not sufficient for the above aim; quantifications of all types of rate constants and those of energy levels, including higher-lying states relevant to the emission process, are required for the complete understanding of the emission mechanism. Recently, we reported rate-constant predictions enabled by a proposed cost-effective calculation method based on Fermi\u2019s golden rule45. Using the method, we calculated kIC, kISC, kRISC, kF, and the rate constant for phosphorescence (kPhos), of benzophenone as an example. The calculated rate constants agreed well with experimental values. Here, we applied the same method to DABNA-1 and determined all relevant rate constants. We also determined the lifetime of the delayed component in transient PL (trPL), the rate constant of TADF (kTADF), and the PLQY, by calculating the population kinetics of the excited states and the trPL decay curve. The calculations also indicated that, after photoexcitation, T1 was first generated via S1\u2009\u2192\u2009T2 ISC and T2\u2009\u2192\u2009T1 IC. Then, triplet-to-singlet conversion occurred via thermally activated T1\u2009\u2192\u2009T2 IC and T2\u2009\u2192\u2009S1 RISC.Here, we report a comprehensive understanding of the TADF mechanism using the representative MR molecule, DABNA-1. So far, \u22060, S1, T1, and T2 geometries of DABNA-1 were obtained at the TPSSh/6-31+G(d) level. Then, excited states were computed using the equation-of-motion coupled-cluster singles and doubles (EOM-CCSD) method (see \u201cMethods\u201d section). At the TD-TPSSh level, S1 and T1 local-energy minima were located at the S1- and T1-optimized geometries, respectively. However, as shown in Supplementary Table\u00a01 and T1 energy levels calculated at the EOM-CCSD level were both located at the T1 geometry . In addition, the S1 and T1 structures were nearly the same, including the dihedral angles . Because S2 states were 0.89\u2009eV higher in energy than the S1 states, their contributions were neglected and that from T2 to T3 was smaller than that from T2 to T1 was only 0.06\u2009eV larger than the experimentally obtained 0.2\u2009eV. Thus, EOM-CCSD significantly improved the overestimation of \u2206E(T1\u2009\u2192\u2009S1), relative to those by previously reported TD-DFT methods44.S1. Figure\u00a0kIC, kISC, kRISC, kF, and kPhos. The raw rate constants in Table\u00a0kIC, kISC, kRISC, kF, and kPhos are given in Supplementary Equations\u00a0We examined triplet formation from SkF(S1\u2009\u2192\u2009S0) (1.4\u2009\u00d7\u2009108\u2009s\u22121) and kIC(S1\u2009\u2192\u2009S0) (1.2\u2009\u00d7\u2009107\u2009s\u22121) agreed quantitatively with experimental results determined by Hatakeyama et al.10 from PL decay curves of a 1-wt% DABNA-1: 9,9\u2032-biphenyl-3,3\u2032-diylbis-9H-carbazole film at 300\u2009K agreed with kISC(S1\u2009\u2192\u2009T2) (5.4\u2009\u00d7\u2009106\u2009s\u22121), rather than kISC(S1\u2009\u2192\u2009T1) (1.7\u2009\u00d7\u2009105\u2009s\u22121), suggesting that the experimental kISC should be assigned to kISC(S1\u2009\u2192\u2009T2). The kISC(S1\u2009\u2192\u2009T2) was ten times greater than kISC(S1\u2009\u2192\u2009T1), because of the larger S1\u2013T2 SOC (1.52\u2009cm\u22121) relative to that of S1\u2013T1 (0.06\u2009cm\u22121). The large S1\u2009\u2192\u2009T2 SOC enhanced S1\u2009\u2192\u2009T2 ISC, despite the uphill transition from S1 to T2 . The small S1\u2013T1 SOC resulted from very similar S1 and T1 orbital configurations (HOMO\u2009\u2192\u2009LUMO transition). These results suggested that the ISC (S1\u2009\u2192\u2009T1 conversion) of DABNA-1 occurred via the stepwise S1\u2009\u2192\u2009T2\u2009\u2192\u2009T1 process, rather than by the one-step S1\u2009\u2192\u2009T1 ISC, irrespective of the geometry relaxation is sufficiently small46. Because T2\u2009\u2192\u2009T1 IC was much faster than S1\u2009\u2192\u2009T2 ISC, the latter was the rate-determining step of the S1\u2009\u2192\u2009T2\u2009\u2192\u2009T1 process was approximately equal to kISC (S1\u2009\u2192\u2009T2)\u2009=\u20095.4\u2009\u00d7\u2009106\u2009s\u22121 (2.5\u2009s\u22121) was considerably less than the experimental value (1\u2009\u00d7\u2009104\u2009s\u22121)10, indicating that T1\u2009\u2192\u2009S1 conversion did not occur in one-step but via a more effective process. Because the S2 and T3 energy levels, as well as higher singlet and triplet states, could not contribute to the RISC from T1, as discussed above, and T2 was close to S1 , we calculated the trPL decay curve were then obtained from the trPL decay curve were calculated fromISC) was calculated fromFor RISC from TT1\u2009\u2192\u2009S1) .5\u2009s\u22121 waT1\u2009\u2192\u2009S1) .5\u2009s\u22121 wakTADF was determined from10Then, k(T1\u2009\u2192\u2009T2\u2009\u2192\u2009S1) was calculated fromFinally, P, \u03a6TADF, \u03a6ISC, \u03c4TADF, kTADF, k(T1\u2009\u2192\u2009T2\u2009\u2192\u2009S1), and k(S1\u2009\u2192\u2009T2\u2009\u2192\u2009T1) values. The \u03a6, \u03a6P, \u03a6TADF, and \u03a6ISC values were quantitatively consistent with the experimental values because of the quantitative predictions for kF(S1\u2009\u2192\u2009S0), kIC(S1\u2009\u2192\u2009S0), kISC(S1\u2009\u2192\u2009T2), and kISC(S1\u2009\u2192\u2009T1). The calculated conversion rate constant k(S1\u2009\u2192\u2009T2\u2009\u2192\u2009T1) was consistent with the experimental 1\u2009\u2192\u2009T2\u2009\u2192\u2009T1 ISC rather than direct S1\u2009\u2192\u2009T1 ISC. All the calculated single-step rate constants and k(S1\u2009\u2192\u2009T2\u2009\u2192\u2009T1) agreed well with the experimental values. However, the agreements of the calculated kTADF and k(T1\u2009\u2192\u2009T2\u2009\u2192\u2009S1) and the experiments were not as good as we expected, compared with the case of k(S1\u2009\u2192\u2009T2\u2009\u2192\u2009T1). Considering that they included the uphill-energy T1\u2009\u2192\u2009T2 IC, there was a possibility that the overestimation of\u00a0\u2206E(T1\u2009\u2192\u2009T2) leads to an underestimation of kIC(T1\u2009\u2192\u2009T2), kTADF, and k(T1\u2009\u2192\u2009T2\u2009\u2192\u2009S1).Table\u00a0E(T1\u2009\u2192\u2009T2) on kIC(T1\u2009\u2192\u2009T2), kTADF, and k(T1\u2009\u2192\u2009T2\u2009\u2192\u2009S1), we recalculated these rate constants using a corrected \u2206E(T1\u2009\u2192\u2009T2). The rate-determining step for the T2-mediated RISC was T1\u2009\u2192\u2009T2 IC; hence, \u2206E(T1\u2009\u2192\u2009T2) could be viewed as the activation energy for DABNA-1 TADF and RISC. As discussed above, the energy difference between the calculated and experimental T1\u2009\u2192\u2009S1 was 60\u2009meV. Thus, 60\u2009meV was subtracted from the EOM-CCSD-calculated \u2206E(T1\u2009\u2192\u2009T2) value of 252\u2009meV. The energy levels and rate constants calculated with the corrected \u2206E(T1\u2009\u2192\u2009T2) are also listed in Table\u00a0k(S1\u2009\u2192\u2009T2\u2009\u2192\u2009T1) was unchanged, suggesting that the T1 energy correction had little effect on the ISC. In contrast, kIC(T1\u2009\u2192\u2009T2) increased from 1.1\u2009\u00d7\u2009107\u2009s\u22121 to 3.3\u2009\u00d7\u2009108\u2009s\u22121. As a result, kTADF and k(T1\u2009\u2192\u2009T2\u2009\u2192\u2009S1) increased to 1.8\u2009\u00d7\u2009104\u2009s\u22121 and 1.9\u2009\u00d7\u2009104\u2009s\u22121, respectively, which were in much better agreements with the experiments. These results suggested that the \u2206E(T1\u2009\u2192\u2009T2) overestimation was responsible for the underestimation of kTADF and k(T1\u2009\u2192\u2009T2\u2009\u2192\u2009S1). Thus, kTADF and k(T1\u2009\u2192\u2009T2\u2009\u2192\u2009S1) depended largely on kIC(T1\u2009\u2192\u2009T2), which was the sum of IC rate constants for individual molecular vibrations, k\u03b1IC,(T1\u2009\u2192\u2009T2), where \u03b1 denoted the \u03b1th vibrational mode (Supplementary Equations\u00a0\u22121) had the largest k\u03b1IC,(T1\u2009\u2192\u2009T2) value method, whereas those of S1, T1, and T2 were performed using the TD-TPSSh/6-31+G(d) method with spin multiplicity\u2009=\u20091 -PCM model reproduced experimental DABNA-1 emission and absorption wavelengths in CH2Cl241. The\u00a0geometry optimization and frequency analysis were performed with the Gaussian 16 program package47. For MR molecules, EOM-CCSD method and algebraic diagrammatic construction of the second order, have been shown to be suitable for calculating singlet\u2013triplet energy differences50. Here, excited-state calculations were performed using the EOM-CCSD/6-31\u2009G method implemented in the Q-Chem program package51. Excitation energies, SOCs, vibronic couplings, transition-dipole moments, and permanent dipole moments were calculated using EOM-CCSD wave functions. Examples of Q-Chem input files for running SOC calculations are shown in the Supplementary Methods.Geometry optimization and frequency analysis of SkIC is described elsewhere52 and reviewed in Supplementary Equation\u00a0\u03b1 and each \u03b1 (Supplementary Equation S5). This protocol was also used for calculating the IC rate constants for Mathematical formulation of a method of calculating vibronic couplings and Supplementary InformationPeer Review File"} +{"text": "To the Editor:TP53 encodes the tumour suppressor protein p53, a master regulator of genomic integrity and cell survival, and is the most frequently mutated gene.3\u2010p53m\u2010RA competitively abolished p53\u2010R175H pulldown from cell lysates with N3\u2010p53m\u2010RA\u2010biotin. By contrast, there was almost no pulldown of wild\u2010type p53 (p53\u2010WT) Figure\u00a0. Structu) Figure\u00a0, but the) Figure\u00a0. Moreove) Figure\u00a0. TherefoA Figure\u00a0.3\u2010p53m\u2010RA, indicating successful conjugation for H1299\u2010p53\u2010R175H and SKBR3 cells were 1.06 and 1.28\u00a0\u00b5M, respectively demonstrated a slight difference in dp53m\u2010RA migration relative to Nn Figure\u00a0. Simulatn Figure\u00a0. Indeed,n Figure\u00a0, suggestn Figure\u00a0. Time\u2010con Figure\u00a0. To subsn Figure\u00a0. The haly Figure\u00a0. When thy Figure\u00a0. The expy Figure\u00a0. Furthery Figure\u00a0. Moreovey Figure\u00a0, and dp5y Figure\u00a0. These fWe next examined the biological consequences of dp53m\u2010RA\u2010mediated p53\u2010R175H degradation in cancer cells. dp53m\u2010RA treatment dramatically inhibited the proliferation of p53\u2010R175H\u2010expressing H1299 cells and SKBR3 cells. By contrast, dp53m\u2010RA treatment did not affect the proliferation of p53\u2010WT\u2010expressing H1299 cells and A549 cells Figure\u00a0. dp53m\u2010RTP53 mutation with dominant\u2010negative and oncogenic gain\u2010of\u2010function activities, as a binder in PROTAC design, we report the first selective mutant p53 degrader, dp53m\u2010RA. dp53m\u2010RA degraded p53\u2010R175H but not wild\u2010type p53 or other p53 mutants in a ubiquitin\u2010proteasome\u2010dependent manner. Importantly, dp53m\u2010RA inhibited the proliferation and migration of cancer cells specifically harbouring the p53\u2010R175H mutation (Figure\u00a0In summary, by harnessing an RNA aptamer specifically targeting p53\u2010R175H, the most common n Figure\u00a0, suggestThe authors declare they have no conflicts of interest.Supporting InformationClick here for additional data file."} +{"text": "E. The terminal benzene ring is approximately perpendicular to the central 1,4-di\u00adaza\u00adbutadiene mean plane, forming a dihedral angle of 81.2\u2005(3)\u00b0.The title mol\u00adecule is disposed about a centre of inversion and the conformation about the imine bond is 52H40N2, is disposed about a centre of inversion and the conformation about the imine bond [1.268\u2005(3)\u2005\u00c5] is E. The terminal benzene ring is approximately perpendicular to the central 1,4-di\u00adaza\u00adbutadiene mean plane, forming a dihedral angle of 81.2\u2005(3)\u00b0. Weak C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 [inter-centroid distance = 4.021\u2005(5)\u2005\u00c5] inter\u00adactions help to consolidate the packing.The title compound, C \u00c5] has an anti-disposition and the imine bonds [1.268\u2005(3)\u2005\u00c5] are E-configured. The dihedral angle between the pendent benzene ring and the 1,4-di\u00adaza\u00adbutadiene least-squares plane is 81.2\u2005(3)\u00b0, consistent with an almost perpendicular relationship. In the crystal, C\u2014H\u22ef\u03c0, Table\u00a01x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z.In this study, we designed and synthesized the title compound Fig.\u00a01 as a pot2O led to the corresponding triphenyl-substituted aniline I, New_Global_Publ_Block. DOI: 10.1107/S2414314620005313/tk4062sup3.txtL1-cif. DOI: 1949863CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The main inter\u00admolecular inter\u00adactions in the two title salts are O\u22efH/H\u22efO contacts, as revealed by Hirshfeld surface analyses. 5H5N4O+\u00b7C7H5O6S\u2212\u00b72H2O, (I), and 6-oxo-1,9-di\u00adhydro\u00adpurin-7-ium perchlorate monohydrate, C5H5N4O+\u00b7ClO4\u2212\u00b7H2O, (II), have been synthesized and characterized using single-crystal X-ray diffraction and Hirshfeld analysis. In both salts, the hypoxanthine mol\u00adecule is protonated at the N7 position of the purine ring. In salt (I), the cation and anion are connected through N\u2014H\u22efO inter\u00adactions. The protonated hypoxanthine cations of salt (I) form base pairs with another symmetry-related hypoxanthine cation through N\u2014H\u22efO hydrogen bonds with an R22(8) ring motif, while in salt (II), the hypoxanthine cations are paired through a water mol\u00adecule via N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds with an R33(11) ring motif. The packings within the crystal structures are stabilized by \u03c0\u2013\u03c0 stacking inter\u00adactions in salt (I) and C\u2014O\u22ef\u03c0 inter\u00adactions in salt (II). The combination of several inter\u00adactions leads to the formation of supra\u00admolecular sheets extending parallel to (010) in salts (I) and (II). Hirshfeld surface analysis and fingerprint plots reveal that O\u22efH/H\u22efO contacts play the major role in the crystal packing of each of the salts, with a 54.1% contribution in salt (I) and 62.3% in salt (II).Two salts of 1,9-di\u00adhydro\u00adpurin-6-one (hypoxanthine), namely, 6-oxo-1,9-di\u00adhydro\u00adpurin-7-ium 5-sulfosalicylate dihydrate, C When hypoxanthine inter\u00adacts with strong acids, it becomes protonated at position N7 or N9. A limited number of hypoxanthine salts like hypoxanthine nitrate , a potential oxygen-free radical generator, is a strong agent against cancer cells (Susithra I), and hypoxan\u00adthin\u00adium perchlorate monohydrate, (II), salts and the noncovalent inter\u00adactions that govern their crystal packings.The current article reports the crystal structures of hypoxanthinium 5-sulfosalicylate dihydrate, (2.I) crystallizes with two hypoxanthinium cations (A+ and B+), two 5-sufosalicylate anions and four solvent water mol\u00adecules in the asymmetric unit, as shown in Fig.\u00a01I), the B cation is equally disordered over two sets of sites for atoms C5B/C5C, C6B/C6C and O6B/O6C. Atoms H1B/H1C and H7B/H7C attached to N1B and N7B, respectively, are also disordered. The solvent water mol\u00adecule O3W is also disordered over two positions. Atoms N7A and N7B are protonated, which is confirmed by widening of the C5A\u2014N7A\u2014C8A angle to 107.1\u2005(4)\u00b0 compared to the value of 103.8\u00b0 in the two polymorphic forms of the neutral HX mol\u00adecule \u00b0 and N3B\u2014C4B\u2014C5C\u2014N7B\u00a0= \u2212178.3\u2005(6)\u00b0 are similar to those of the two forms of the neutral HX mol\u00adecule \u00b0 and O7B\u2014C9B\u2014C10B\u2014C11B\u00a0= 175.9\u2005(4)\u00b0], a situation that is likewise observed for previously reported crystal structures involving 5SCA\u2212 anions.Salt (II) crystallizes with one hypoxanthinium cation, one perchlorate anion (PCA\u2212) and one solvent water mol\u00adecule in the asymmetric unit. The mol\u00adecular structure of salt (II) is shown in Fig.\u00a02I). The PCA\u2212 anion has the characteristic tetra\u00adhedral shape, with Cl\u2014O bond lengths between 1.4116\u2005(15) and 1.4421\u2005(15)\u2005\u00c5, and O\u2014Cl\u2014O angles between 108.29\u2005(9) and 111.24\u2005(12)\u00b0.Salt (3.I), (010) sheets of cations and sheets of anions are stacked alternately along [010]. The crystal packing is governed by N\u2014H\u22efO, O\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds self assemble into sheets by inter\u00adaction of symmetry-related counterparts through O7A\u2014H7D\u22efO10A and O7B\u2014H7E\u22efO10B, respectively via N1B\u2014H1C\u22efO11A and C2B\u2014H2B\u22efO11A with an W\u2014H1WA\u22efO10B, O1W\u2014H1WB\u22efO12B, O2W\u2014H2WA\u22efN3A and O2W\u2014H2WB\u22efO12A is consolidated by \u03c0\u2013\u03c0 inter\u00adactions between the phenyl rings of the two 5SCA anions (C10A\u2013C15A and C10B\u2013C15B), and the imidazole ring (C4A\u2013N9A) and the pyrimidine ring (N1A\u2013C6A) of cation A+, with centroid-to-centroid distances of 3.547\u2005(3), 3.562\u2005(3), 3.554\u2005(3) and 3.533\u2005(3)\u2005\u00c5, and slippages of 0.815, 1.300, 1.182 and 1.105\u2005\u00c5 , (010) sheets of cations and sheets of anions are stacked alternately along [010]. The crystal packing of salt (II) is dominated by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, and to a minor extent by C\u2014H\u22efO hydrogen bonds is shown in Fig.\u00a08In the crystal structure of salt (s Table\u00a02. The pro4.I) and (II) were performed using CrystalExplorer17 and 10(b) for (I) and (II), respectively. Corresponding two-dimensional fingerprint plots and (II) are shown in Figs. 11I), 62.3% (II); N\u22efH/H\u22efN 3.1% (I), 6.8% (II); C\u22efH/H\u22efC 5.9% (I), 5.4% (II); H\u22efH/H\u22efH 16.0% (I), 5.3% (II); C\u22efC/C\u22efC 0.9% (I), 0.1% (II).Hirshfeld surface (HS) analyses of salts and (II) with the related salt forms of guanine, xanthinium and hypoxanthine reveal that, in most of the crystal structures containing purine derivatives, the purine forms base pairs through pairs of N\u2014H\u22efO or N\u2014H\u22efN hydrogen bonds with an et al., 2014I) and (II) and related systems compiled in Table\u00a03A comparison of salts (6.I) was synthesized by mixing an equimolar ratio of hypoxanthine (0.0340\u2005g) and 5-sulfosalicylic acid (0.0545\u2005g) in hot water. The solution was heated to 333\u2005K for 1\u2005h and then allowed to cool slowly to room tem\u00adper\u00adature. Colourless needle-shaped crystals were harvested from the mother liquid after one week.Salt (II) was synthesized by mixing an equimolar ratio of hypoxanthine (0.0340\u2005g) and iron perchlorate monohydrate (0.0681\u2005g) in hot water. The solution was heated to 333\u2005K with constant stirring for 1\u2005h and then allowed to cool slowly to room tem\u00adper\u00adature. Colourless plate-like crystals were harvested from the mother liquid after one week.Salt (7.I) and (II) are summarized in Table\u00a04I), carbon (C5 and C6) and oxygen (O6) atoms of cation B are equally disordered over two sets of sites, with a refined occupancy ratio of 0.503\u2005(18):0.497\u2005(18). The solvent water mol\u00adecule O3W is disordered over two positions, with a refined site-occupancy ratio of 0.58\u2005(6):0.42\u2005(6). The H atoms of water mol\u00adecules O1W and O2W were located from a difference Fourier map, and the O\u2014H distance restrained to 0.82\u2005\u00c5. Attempts to localize the H atoms of O3W and O4W in (I) from difference Fourier maps failed as there were no relevant electron densities close to these atoms. Hence, these H atoms are not part of the model but are included in the formula. All C- and N-bound H atoms in (I) were placed in idealized positions and refined freely using a riding model, with C\u2014H\u00a0= 0.95\u2005\u00c5 and N\u2014H\u00a0= 0.86\u2005\u00c5, and with Uiso(H)\u00a0= 1.2Ueq. In salt (II), the N-bound H atoms were located in a difference Fourier map and refined freely. The H atoms of the water mol\u00adecule were likewise located from a difference Fourier map. The geometry of the water mol\u00adecule was restrained using DFIX commands with an O\u2014H distance of 0.85\u2005\u00c5 and an H\u22efH distance of 1.36\u2005\u00c5. All C-bound H atoms were treated as for salt (I).Crystal data, data collection and structure refinement details of salts I, II, global. DOI: 10.1107/S2056989022004753/wm5640Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989022004753/wm5640IIsup3.hklStructure factors: contains datablock(s) II. DOI: 2170315, 2170314CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The 1,3-oxazolidine ring adopts an envelope conformation with the N atom in an endo position. The mean plane of the oxazolidine ring makes dihedral angles of 77.74\u2005(10) and 45.50\u2005(11)\u00b0, respectively, with the 4-bromo\u00adphenol and 1,3,5-tri\u00admethyl\u00adbenzene rings. In the crystal, adjacent mol\u00adecules are connected via C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions into layers parallel to the (200) plane. The packing is strengthened by van der Waals inter\u00adactions between parallel mol\u00adecular layers. A Hirshfeld surface analysis shows that H\u22efH (58.2%), C\u22efH/H\u22efC (18.9%), and Br\u22efH/H\u22efBr (11.5%) inter\u00adactions are the most abundant in the crystal packing.The title compound, C The mean plane of th oxazolidine ring makes dihedral angles of 77.74\u2005(10) and 45.50\u2005(11)\u00b0, respectively, with the 4-bromo\u00adphenol (C6\u2013C11) and the 1,3,5-tri\u00admethyl\u00adbenzene (C14\u2013C19) rings. The mol\u00adecular conformation is stabilized by intra\u00admolecular O11\u2014H11\u22efN3 and C20\u2014H20C\u22efO1 hydrogen bonds plane , C\u22efH/H\u22efC (18.9%), and Br\u22efH/H\u22efBr (11.5%) contacts are shown in Fig.\u00a05b\u2013d, while numerical details of the different contacts are given in Table\u00a02A Hirshfeld surface analysis was performed and the associated two-dimensional fingerprint plots were obtained with 4.et al., 2016S)-5-chloro-N-({2-oxo-3-[4-(3-oxomorpholin-4-yl)phen\u00adyl]oxa\u00adzolidin-5-yl}meth\u00adyl)-thio\u00adphene-2-carboxamide [(I): Shen et al., 2018et al., 2010et al., 2010et al., 2012R)-2-phen\u00adoxy-1-ethanone [(V): Caracelli et al., 2011A search of the Cambridge Structural Database , adjacent mol\u00adecules are connected via O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions into a zigzag chain along the b-axis direction. In (V), mol\u00adecules are linked into supra\u00admolecular arrays two mol\u00adecules thick in the bc plane through C\u2014H\u22efO, C\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions.In the crystal of (I)5.The title compound was synthesized using our recently reported procedure = 1.2 or 1.5Ueq(C). The hydroxyl H atom was found in a difference-Fourier map and was refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022005928/tx2051sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022005928/tx2051Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022005928/tx2051Isup3.cmlSupporting information file. DOI: 2176709CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The first crystal structure is reported of a mol\u00adecule containing the 1,4,2-di\u00adthia\u00adzole-5-thione heterocycle. The packing features aromatic \u03c0\u2013\u03c0 stacking, weak C\u2014H\u22efS inter\u00adactions and short S\u22efS inter\u00adactions. 8H5NS3, the dihedral angle between the heterocyclic ring and the phenyl ring is 2.62\u2005(5)\u00b0. In the extended structure, aromatic \u03c0\u2013\u03c0 stacking between the 1,4,2-di\u00adthia\u00adzole-5-thione moiety and the phenyl ring is observed [centroid\u2013centroid distances = 3.717\u2005(6) and 3.712\u2005(6)\u2005\u00c5]. The almost planar mol\u00adecules arrange themselves in parallel chains of head-to-tail mol\u00adecules oriented by a network of weak C\u2014H\u22efS contacts close to the sum of their van der Waals radii within the chains. All the hydrogen atoms participate in hydrogen-bonding inter\u00adactions with the sulfur and nitro\u00adgen atoms of adjacent mol\u00adecules. C=S\u22efS contacts between the chains that are significantly shorter than the sum of their van der Waals radii also impact the overall packing.In the title compound, C This pattern of bond lengths is in agreement with a more extensive, and less localized, \u03c0 delocalization in this heterocycle than in the comparable oxa\u00adthia\u00adzolone derivatives.Within the heterocycle moiety, the mol\u00adecule shows significant \u00b0] on the a axis. Head-to-toe hydrogen-bonding inter\u00adactions between C5 and C6 donors to the exocyclic thione S3 with H\u22efS distances of 2.96 and 3.11\u2005\u00c5, respectively , H5\u22efS1 (3.04\u2005\u00c5), H7\u22efS3 (3.06\u2005\u00c5) and H8\u22efS2 (3.10\u2005\u00c5). The sulfur\u2013sulfur inter\u00adactions occur as a chain out of plane between the thione S3 and S1 atoms within the ring = 0.671; UV\u2013visible (CH2Cl2) \u03bbmax nm (log \u025b): 256 (4.29), 361 (4.20), 1H NMR 60\u2005MHz, CDCl3) \u03b4 = 7.53 ppm (multiplet).A solution of tri\u00adchloro\u00admethane\u00adsulfenyl chloride in chloro\u00adform (10.17\u2005g) was added dropwise to a warmed solution of thio\u00adbenzamide in chloro\u00adform (240\u2005ml) according to a literature procedure (Greig 6.Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698902200888X/hb8037sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902200888X/hb8037Isup2.hklStructure factors: contains datablock(s) I. DOI: 2205416CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular structure features an angular (phenyl\u00adiodos\u00adyl)benzene cation, the geometry of which was hitherto undescribed in the literature: in the cation, both I\u2014C bonds are approximately normal to the I=O bond, forming a C\u2014I\u2014C angle of 95.36\u2005(4)\u00b0. The crystal structure displays O\u2014H\u22efO, O\u2014H\u22efI and O\u2014H\u22efS hydrogen bonding.The structure of the title salt (systematic name: oxodiphenyl-\u03bb This iodosyl salt arose from the reaction of iodoxybenzene with sodium hydroxide benzene tosyl\u00adate dihydrate \u00b0. The bond lengths of C1\u2014I1 and C7\u2014I1 are 2.1289\u2005(11) and 2.1370\u2005(12)\u2005\u00c5, respectively. These values are comparable to the sum of the van der Waals radii -one, i.e. the Dess\u2013Martin periodinane, with a C\u2014I bond length between the phenyl and iodine of 2.1025\u2005(16)\u2005\u00c5 \u2005\u00c5], resides nearly perpendicular at 77.55\u2005(4)\u00b0 to the I1\u2014O1 bond. This bond length is shorter than the sum (3.05\u2005\u00c5) of the covalent radii, and this is analogous to secondary bonding that was observed by Rentzeperis in his bis\u00ad(di\u00adphenyl\u00adiodo\u00adnium I-oxide) di\u00adacetate trihydrate, between the acetate anion and the iodo\u00adnium I-oxide centre di\u00adacetate trihydrate and 2.8118\u2005(10)\u2005\u00c5, respectively.Zhdankin synthesized a tosyl\u00adate derivative of 2-iodoxybenzoic acid. Crystals of the final product could not be isolated, but the inter\u00admediate mixed tosyl\u00adate-acetate derivative was analysed. The I\u2014O bond lengths in this iodine(V) compound had distances of 2.080\u2005(2), 2.213\u2005(2), 2.027\u2005(2) and 1.998\u2005(2)\u2005\u00c5 (Yusubov I-oxide) di\u00adacetate trihydrate also adopted a distorted octa\u00adhedral geometry, albeit via a dimeric coordinating structure \u00b0, the C7\u2014I1\u2014O3 angle of 176.33\u2005(4)\u00b0, the O1\u2014I1\u2014O4 angle of 77.55\u2005(4)\u00b0 with the coordinating tosyl\u00adate anion and the C1\u2014I1\u2014C7 angle of 95.36\u2005(4)\u00b0 complete the distorted octa\u00adhedral geometry. The accompanying tosyl\u00adate anion and water mol\u00adecules occupy apical and equatorial positions to stabilize the monomeric complex. Bis and 1.99\u2005(2)\u2005\u00c5, respectively, as viewed down the a axis di\u00adacetate trihydrate was added to the cooled filtrate and a white precipitate formed. The suspension was stirred for an additional 30\u2005min and then filtered. The compound was washed with a minimal amount of diethyl ether (10\u2005ml) and ice-cold water (10\u2005ml). The product matched known 1H, 13C and FTIR data benzene tosyl\u00adate dihydrate was synthesized according to a modified procedure by Chen 2007 and is iCrystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314622005223/bh4069sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2414314622005223/bh4069Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622005223/bh4069Isup3.cmlSupporting information file. DOI: 2173122CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "At 293\u2005K, [tBuNHC2Me][Br]2\u00b7H2O crystallizes in the P21/c space group, while [tBuNHC2Et][Br]2\u00b72H2O crystallizes in the P21/n space group at 100\u2005K. At 112\u2005K, [MesNHC2Me][Br]2\u00b72H2O crystallizes in the ortho\u00adrhom\u00adbic space group Pccn while [MesNHC2Et][Br]2\u00b74H2O crystallizes in the P21/c space group at 100\u2005K.The crystal structures of ligand precursor bis\u00ad(imidaozolium) salts 1,1\u2032-methyl\u00adenebis(3- tert-butyl\u00adimidazolium) dibromide monohydrate, C15H26N4+\u00b72Br\u2212\u00b7H2O or [tBuNHC2Me][Br]2\u00b7H2O, 1,1\u2032-bis\u00ad(3-tert-butyl\u00adimidazolium) dibromide dihydrate, C16H28N4+\u00b72Br\u2212\u00b72H2O or [tBuNHC2Et][Br]2\u00b72H2O, 1,1\u2032-methyl\u00adenebis[3-imidazolium] dibromide dihydrate, C25H30N42+\u00b72Br\u2212\u00b72H2O or [MesNHC2Me][Br]2\u00b72H2O, and 1,1\u2032-bis\u00ad[3-imidazolium] dibromide tetra\u00adhydrate, C26H32N42+\u00b72Br\u2212\u00b74H2O or [MesNHC2Et][Br]2\u00b74H2O, are reported. At 293\u2005K, [tBuNHC2Me][Br]2\u00b7H2O crystallizes in the P21/c space group, while [tBuNHC2Et][Br]2\u00b72H2O crystallizes in the P21/n space group at 100\u2005K. At 112\u2005K, [MesNHC2Me][Br]2\u00b72H2O crystallizes in the ortho\u00adrhom\u00adbic space group Pccn while [MesNHC2Et][Br]2\u00b74H2O crystallizes in the P21/c space group at 100\u2005K. Bond distances and angles within the imidazolium rings are generally comparable among the four structures. All four bis\u00ad(imidazolium) salts co-crystallize with one to four mol\u00adecules of water.The crystal structures of ligand precursor bis\u00ad(imidazolium) salts 1,1\u2032-methyl\u00adenebis(3- Bis(imidazolium) salts, [RNHC2R1][X]2 are relatively modular in that modifications can be relatively easily made to exterior groups attached to each NHC (R), the moiety linking the two NHC groups (R1), and the counter-ion (X). One general synthetic approach for synthesizing bis(imidazolium) salts is where two equivalents of an alkyl or aryl imidazole are combined with one equivalent of an organic dihalide reagent and refluxed to afford the final product salts can be relatively easily synthesized from alkyl or aryl imidazoles, and some are also commercially available.Bis(imidazolium) salts are common precursors for the synthesis of bidentate tert-butyl (tBu) and mesityl (Mes) exterior R groups and methyl\u00adene (Me) or ethyl\u00adene (Et) linking R1 groups. [MesNHC2Et][Br]2 was even reported to be a stand-alone catalyst for the conversion of aryl\u00adaldehydes to carb\u00adoxy\u00adlic acids in combination with water and K2CO3 in DMSO were reported to be more effective for hydro\u00adsilylation reactions with RhI complexes than ethyl\u00adene linkers salts are those with tBuNHC2Me][Br]2 and [tBuNHC2Et][Br]2 were used as precursors for synthesis of rhodium complexes 2-assisted deprotometalation salts with tert-butyl and mesityl ancillary groups.While bis\u00ad(imidazolium) salts are common ligand precursors, few have been structurally characterized salts were recrystallized from hot methanol and each compound co-crystallizes with one or more mol\u00adecules of water. Fig.\u00a01tBuNHC2Me][Br]2\u00b7H2O and [tBuNHC2Et][Br]2\u00b72H2O are mostly the same within experimental error, with backbone C2\u2014C3 distances of 1.348\u2005(4) and 1.349\u2005(3)\u2005\u00c5, respectively. The N\u2014C distances are also mostly comparable with [tBuNHC2Me][Br]2\u00b7H2O having an N1\u2014C2 and an N2\u2014C3 distance of 1.389\u2005(3)\u2005\u00c5 and N1\u2014C1 and N2\u2014C1 distances both being 1.337\u2005(3)\u2005\u00c5, while [tBuNHC2Et][Br]2\u00b72H2O has an N1\u2014C2 distance of 1.388\u2005(3)\u2005\u00c5, an N2\u2014C3 distance of 1.384\u2005(3)\u2005\u00c5, an N1\u2014C1 distance of 1.327\u2005(3)\u2005\u00c5 and an N2\u2014C1 distance of 1.331\u2005(3)\u2005\u00c5. For the linker, the N2\u2014C7 distance is 1.463\u2005(3)\u2005\u00c5 for [tBuNHC2Me][Br]2\u00b7H2O and 1.468\u2005(3)\u2005\u00c5 for [tBuNHC2Et][Br]2\u00b72H2O.Bond distances in the imidazolium rings of [tBuNHC2Me][Br]2\u00b7H2O and [tBuNHC2Et][Br]2\u00b72H2O. For [tBuNHC2Me][Br]2\u00b7H2O, bond angles include C1\u2014N1\u2014C2 at 108.2\u2005(2)\u00b0, N1\u2014C2\u2014C3 at 107.6\u2005(2)\u00b0, C2\u2014C3\u2014N2 at 106.9\u2005(2)\u00b0, C3\u2014N2\u2014C1 at 108.6\u2005(2)\u00b0, and N2\u2014C1\u2014N1 at 108.7\u2005(2)\u00b0. For [tBuNHC2Et][Br]2\u00b7H2O, bond angles include C1\u2014N1\u2014C2 at 108.21\u2005(19)\u00b0, N1\u2014C2\u2014C3 at 107.3\u2005(2)\u00b0, C2\u2014C3\u2014N2 at 106.9\u2005(2)\u00b0, C3\u2014N2\u2014C1 at 108.54\u2005(19)\u00b0, and N2\u2014C1\u2014N1 at 109.02\u2005(19)\u00b0.Bond angles in the imidazolium rings are also quite similar in [MesNHC2Me][Br]2\u00b72H2O while Fig.\u00a04MesNHC2Et][Br]2\u00b74H2O. Notably, [MesNHC2Et][Br]2\u00b74H2O is the only compound of the four for which the asymmetric unit contains only half of the mol\u00adecule.Fig.\u00a03MesNHC2Me][Br]2\u00b72H2O and [MesNHC2Et][Br]2\u00b74H2O are mostly the same within experimental error, with backbone C2\u2014C3 distances of 1.344\u2005(3) and 1.3506\u2005(19)\u2005\u00c5, respectively. N\u2014C distances are also mostly the same with [MesNHC2Me][Br]2\u00b72H2O having an N1\u2014C2 distance of 1.387\u2005(3)\u2005\u00c5, an N2\u2014C3 distance of 1.380\u2005(3)\u2005\u00c5, an N1\u2014C1 distance of 1.326\u2005(3)\u2005\u00c5, and an N2\u2014C1 distance of 1.341\u2005(3)\u2005\u00c5. Similarly, [MesNHC2Et][Br]2\u00b74H2O has an N1\u2014C2 distance of 1.3872\u2005(16)\u2005\u00c5, an N2\u2014C3 distance of 1.3841\u2005(16)\u2005\u00c5, an N1\u2014C1 distance of 1.3322\u2005(16)\u2005\u00c5 and an N2\u2014C1 distance of 1.3314\u2005(16)\u2005\u00c5. For the linker, the N2\u2014C7 distance is 1.457\u2005(3)\u2005\u00c5 for [MesNHC2Me][Br]2\u00b72H2O and 1.4653\u2005(16)\u2005\u00c5 for [MesNHC2Et][Br]2\u00b74H2O.Bond distances in the imidazolium rings of [MesNHC2Me][Br]2\u00b72H2O and [MesNHC2Et][Br]2\u00b74H2O. For [MesNHC2Me][Br]2\u00b72H2O, bond angles include C1\u2014N1\u2014C2 at 108.92\u2005(17)\u00b0, N1\u2014C2\u2014C3 at 107.20\u2005(19)\u00b0, C2\u2014C3\u2014N2 at 106.95\u2005(19)\u00b0, C3\u2014N2\u2014C1 at 108.96\u2005(17)\u00b0, and N2\u2014C1\u2014N1 at 107.96\u2005(18)\u00b0. For [MesNHC2Et][Br]2\u00b74H2O, bond angles include C1\u2014N1\u2014C2 at 108.51\u2005(11)\u00b0, N1\u2014C2\u2014C3 at 107.19\u2005(11)\u00b0, C2\u2014C3\u2014N2 at 106.87\u2005(11)\u00b0, C3\u2014N2\u2014C1 at 108.89\u2005(11)\u00b0, and N2\u2014C1\u2014N1 at 108.54\u2005(11)\u00b0. Overall, these data support that changing the linker R1 group from methyl\u00adene to ethyl\u00adene does not significantly affect the imidazolium ring structures.Bond angles in the imidazolium rings are also mostly the same for [3.tBuNHC2Me][Br]2\u00b7H2O is stabilized by hydrogen bonding \u2005\u00c5. One tert-butyl group has positional disorder.The supra\u00admolecular structure of [ng Fig.\u00a05. Distancng Fig.\u00a05. HydrogetBuNHC2Et][Br]2\u00b72H2O is stabilized by extensive hydrogen bonding and Br1\u22efH1A (2.398\u2005\u00c5).The supra\u00admolecular structure of [ng Fig.\u00a06. Distancng Fig.\u00a06. SeveralMesNHC2Me][Br]2\u00b72H2O is also stabilized by hydrogen bonding \u2005\u00c5, O2\u22efH1E at 2.001\u2005(3)\u2005\u00c5, and Br1\u22efH1D at 2.585\u2005(2)\u2005\u00c5.The supra\u00admolecular structure of [Br]2\u00b7H2O, [tBuNHC2Et][Br]2\u00b72H2O, or [MesNHC2Et][Br]2\u00b74H2O. A deposited dataset for [MesNHC2Me][Br]2\u00b72H2O was found . The procedure was adapted from a literature procedure , approximately 100\u2005mL of methanol, and approximately 25\u2005mL of deionized water and a stir bar, then heated to 343\u2005K under reflux. 7.81\u2005mL of 37% formaldehyde were added, followed by 3.70\u2005mL of ammonium hydroxide added dropwise over 5 minutes while stirring. The solution was refluxed at 343\u2005K for 5\u2005h, resulting in a light red\u2013orange solution. Excess solvent was removed in vacuo, and the resulting product was diluted with approximately 150\u2005mL of di\u00adchloro\u00admethane and washed twice with 50\u2005mL of deionized H2O until the aqueous layers ran clear. The product was vacuum distilled at \u223c373\u2005K, yielding a clear liquid, which was weighed in a tared vial, resulting in 7.95\u2005g (34% yield) of tBuIm, and characterized by 1H NMR spectroscopy in CDCl3.Synthesis of 1--1HMesIm).-imidazole, of tan\u2005crystals, which were characterized by 1H NMR spectroscopy and identified as MesIm.Synthesis of 1,1\u2032-di(tert-but\u00adyl)-3,3\u2032-methyl\u00adene-diimidazolium dibromide, [tBuNHC2Me][Br]2. 1.850\u2005g of tbuIm and 0.4194\u2005mL of di\u00adbromo\u00admethane, a stir bar, and \u223c20\u2005mL of toluene were stirred in a 50\u2005mL round-bottomed flask. The solution was then heated to 423\u2005K and refluxed for 46\u2005h, resulting in the formation of a dark orange\u2013brown solution. The solution was cooled in an ice bath, resulting in a fine white precipitate which was collected via vacuum filtration, washed twice with \u223c5\u2005mL of cold toluene, filtered and dried. 1.120\u2005g (78.02% yield) of a fine white solid identified as [tBuNHC2Me][Br]2 were isolated. Crystals suitable for X-ray diffraction were obtained by recrystallization from hot methanol. The product was characterized by 1H NMR spectroscopy. The 1H NMR data were consistent with those previously reported -3,3\u2032-ethyl\u00adene-diimidazolium dibromide [tBuNHC2Et][Br]2. A 250\u2005mL round-bottomed flask was charged with 2.017\u2005g of tbuIm, 0.562\u2005mL of di\u00adbromo\u00adethane, a stir bar, and \u223c20\u2005mL of toluene. The mixture was refluxed at 423\u2005K and stirred for 46\u2005h, at which point the solution was a rusty brown color. The flask was then placed in an ice bath, and the resulting precipitate was collected via vacuum filtration and washed twice with \u223c5\u2005mL of cold toluene. The resulting solids were dried and weighed, yielding 1.727\u2005g (61% yield) of [tBuNHC2Et][Br]2 and single crystals suitable for X-ray diffraction were obtained via recrystallization from hot methanol. 1H NMR data were consistent with those previously reported -3,3\u2032-methyl\u00adene-diimidazolium dibromide, [Br]2 suitable for X-ray diffraction and characterized by 1H NMR.MesNHC2Et][Br]2.Synthesis of 1,1\u2032-di(mesit\u00adyl)-3,3\u2032-ethyl\u00adene-diimidazolium dibromide, [Br]2.6.SHELXL and included as riding contributions with distances of 0.95\u2005\u00c5 for C\u2014H, 0.99\u2005\u00c5 for CH2 and 0.98\u2005\u00c5 for CH3. Methyl H atoms were allowed to rotate but not to tip to best fit the experimental electron density. Uiso values of riding H atoms were set to 1.2 times Ueq(C) for CH and CH2, and 1.5 times Ueq(C) for CH3 and H2O. For [tBuNHC2Me][Br]2, the SADI command of SHELX was used to model disorder in one of the tert-butyl moieties for N4\u2014C0AA and N4\u2014C12, C0AA\u2014C00N and C14\u2014C12, and C1AA\u2014C0AA and C13\u2014C12 to restrain distances within a sigma of 0.02\u2005\u00c5. The population parameters for the disordered tert-butyl groups are 0.54019 for C12\u2013C14, and 0.45981 for C00N, C0AA, and C1AA. The highest peak and deepest hole are both near a heavy atom Br1 with a distance of 0.88\u2005\u00c5 from the highest peak of 1.49\u2005e\u2005\u00c5\u22123 and a distance of 0.73\u2005\u00c5 from the deepest hole of \u22121.10\u2005e\u2005\u00c5\u22123.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022008003/mw2188sup1.cifCrystal structure: contains datablock(s) sces01006_0m, est01043_0m, at01019_0ma, est01041d_0ma, global. DOI: 10.1107/S2056989022008003/mw2188sces01006_0msup2.hklStructure factors: contains datablock(s) sces01006_0m. DOI: 10.1107/S2056989022008003/mw2188est01043_0msup3.hklStructure factors: contains datablock(s) est01043_0m. DOI: 10.1107/S2056989022008003/mw2188at01019_0masup4.hklStructure factors: contains datablock(s) at01019_0ma. DOI: 10.1107/S2056989022008003/mw2188est01041d_0masup5.hklStructure factors: contains datablock(s) est01041d_0ma. DOI: Click here for additional data file.10.1107/S2056989022008003/mw2188sces01006_0msup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022008003/mw2188est01043_0msup7.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022008003/mw2188at01019_0masup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022008003/mw2188est01041d_0masup9.cmlSupporting information file. DOI: 2195736, 2195735, 2195734, 2195733CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In recent years, a number of mass spectrometric and laser spectroscopic techniques have been developed and are increasingly used by the research community. Advances in this active research area, however, critically depend on the availability of suitable N2O isotope Reference Materials (RMs).Information on the isotopic composition of nitrous oxide have been analysed by specialised laboratories against isotope reference materials. A particular focus was on the 15N site\u2010specific isotopic composition, as this measurand is both highly diagnostic for source appointment and challenging to analyse and link to existing scales.Within the project Metrology for Stable Isotope Reference Standards (SIRS), seven pure N2O isotope RMs offer a wide spread in delta (\u03b4) values: \u03b415N: 0 to +104\u2030, \u03b418O: +39 to +155\u2030, and \u03b415NSP: \u22124 to +20\u2030. Conversion and uncertainty propagation of \u03b415N and \u03b418O to the Air\u2010N2 and VSMOW scales, respectively, provides robust estimates for \u03b415N(N2O) and \u03b418O(N2O), with overall uncertainties of about 0.05\u2030 and 0.15\u2030, respectively. For \u03b415NSP, an offset of >1.5\u2030 compared with earlier calibration approaches was detected, which should be revisited in the future.The established N2O isotope RMs anchored to the international isotope\u2010ratio scales was developed that will promote the implementation of the recommended two\u2010point calibration approach. Particularly, the availability of \u03b417O data for N2O RMs is expected to improve data quality/correction algorithms with respect to \u03b415NSP and \u03b415N analysis by mass spectrometry. We anticipate that the N2O isotope RMs will enhance compatibility between laboratories and accelerate research progress in this emerging field.A set of seven N For nitrogen, the 15N/14N isotope ratio is used, R(15N/14N)\u2009=\u00a0x(15N)/x(14N), where x is the isotopic abundance and tropospheric N2 is the international reference material for the Air\u2010N2 scale. For oxygen, the 18O/16O and 17O/16O ratios are used, which are related to the Vienna Standard Mean Ocean Water (VSMOW) scale. In addition, we adopt the following notation conventions: \u03b415N\u2009=\u00a0\u03b4 (average of both nitrogen atoms) and \u03b418O\u2009=\u00a0\u03b4. The 15N site preference (SP) is defined by the predominance of 15N substitution in the central (\u03b1) position as compared to the terminal (\u03b2) position, and calculated accordingly as \u03b415NSP\u00a0=\u00a0\u03b415N\u03b1\u00a0\u2212\u00a0\u03b415N\u03b2. All \u03b4 values in this paper are reported against Air\u2010N2 (for 15N/14N ratios) and against VSMOW (for 18O/16O and 17O/16O ratios).The isotopic composition of a sample is reported using the delta as well as \u03b417O and \u03b418O (water) have a different chemical identity than N2O sample gas. Thus, a chemical conversion reactionFurther progress in N2O by thermal decomposition of isotopically characterised ammonium nitrate (NH4NO3) has been suggested as an approach to link the position\u2010dependent nitrogen isotopic composition of N2O to the Air\u2010N2 scale.\u03b1\u2010position of of the formed N2O originates from NO3\u2212, while the \u03b2\u2010nitrogen comes from NH4+.4NO3 decomposition technique has been confirmed,\u03b415N\u03b1 and \u03b415N\u03b2 was found to be limited by non\u2010quantitative NH4NO3 decomposition in combination with substantially different isotope enrichment factors of \u22124 or \u221219\u2030 for the conversion of the NO3\u2212 or NH4+ nitrogen atom into the \u03b1\u2010 or \u03b2\u2010position of the N2O molecule.2O reference gases, USGS51 and USGS52, recently became available with assigned \u03b4 values based on a preliminary assessment by Naohiro Yoshida and Sakae Toyoda (Tokyo Institute of Technology).\u03b415N and \u03b418O values (< 1\u2030), which is not suitable for a two\u2010point calibration approach.The synthesis of N2O RMs within the framework of the European Metrology Programme for Innovation and Research (EMPIR) 16ENV06 project \u2018Metrology for Stable Isotope Reference Standards (SIRS)\u2019. The target isotopic composition of N2O RMs was selected according to discussions at a stakeholder workshop at the 19th GGMT conference at Empa (29 August 2017).2O RMs compared to RMs presented in Ostrom et al\u03b417O data in order to improve data quality/correction algorithms with respect to \u03b415NSP and \u03b415N analysis by mass spectrometry. In addition, the link of \u03b4 values to the international isotope\u2010ratio scales was revisited.In the present study, we report the development of additional N22O RMs, covering an extended range of delta values as compared to existing gases. Figure\u00a02O RMs.The main purpose of this study is the provision of isotopically characterised N15N/14N isotope ratios on the Air\u2010N2 scale were propagated from NH4+ and NO3\u2212 salts supplied by IAEA/USGS, through isotopic analysis of gravimetrically prepared NH4NO3 salts (section\u00a0\u03b415N\u03b2(N2O) /\u03b415N\u03b1(N2O) in the novel N2O RMs. The international RMs applied in this study are listed in Table\u00a02O site\u2010specific isotopic composition, the NH4NO3 decomposition reaction was optimised for high yield, reproducibility, and N2O purity , \u03b418O(N2O), \u03b417O(N2O) and \u03b415NSP(N2O) is described. In one laboratory (Empa), \u03b415NSP(N2O) in the N2O RMs was linked to the Air\u2010N2 scale making use of the traceability chain established in section\u00a0In section\u00a02.12.1.14NO3 salts (S1\u2013S6), covering a wide range of \u03b415N(NH4+) and \u03b415N(NO3\u2212) values, were produced by gravimetric mixing of five commercially available NH4NO3 salts (A\u2013E). A: unlabelled NH4NO3 , B: 15NH4NO3 , C: NH415NO3 , D: 15NH4+\u2010depleted NH4NO3 , E: 15NO3\u2212\u2010depleted NH4NO3 .Six NH4NO3 salts (Table\u00a04NO3 (A) was ground to a fine powder using a mortar and pestle and then dried at 120\u00b0C for 1\u00a0h (a temperature low enough to avoid triggering decomposition). From this, around 100\u2009g (S1\u2013S5) or around 40\u2009g (S6) were gravimetrically mixed with appropriate amounts of salts B, C, D, and E to obtain the desired isotopic composition. The salt mixtures were dissolved in deionised water , recrystallised, dried, and then stored in air\u2010tight sample containers. The isotopic homogeneity of S1\u2013S6 was confirmed by repeated IRMS analysis (MPI\u2010BGC), demonstrating \u03b415N(NH4NO3) values within <0.2\u2030 .For preparation of these six NHts Table\u00a0, approxi2.1.24NO3 salts (S1\u2013S6) were sent together with international reference materials ((NH4)2SO4, NaNO3, KNO3) provided by the IAEA and by USGS Table\u00a0.\u03b415N(NH4NO3), \u03b415N(NH4+) and \u03b415N(NO3\u2212) results from all laboratories were calibrated using the provided international IAEA and USGS reference materials, with \u03b415N values and uncertainties according to Brand et al\u03c3cal) was estimated from the uncertainty in the linear calibration function (Equation\u00a0\u03c3meas) in \u03b415Nmeas, following the law of error propagation from individual laboratories i were combined to a weighted mean value were weighed into round\u2010bottomed glass flasks with a break\u2010seal . In a variant of the NH4NO3 decomposition reaction according to Szab\u00f3 et al,4HSO4 and 0.2\u00a0g (NH4)2SO4 were added. Adding surplus NH4+ salt will lead to a loss in \u03b415N\u03b2 information but was included to test if very high reaction yields can be achieved, which might still be attractive. Therefore, for S1, both variants (with/without NH4HSO4/(NH4)2SO4) were tested, while for S2\u2013S6 only decomposition without NH4+ addition was performed. Thereafter, the flasks were evacuated (<10\u22121\u00a0mbar) and flame\u2010sealed. The sealed flasks were placed in a circulating\u2010air oven and heated to 270\u00b0C for 24\u2009h.Aliquots of approximately 1.0\u00a0g (12.5\u00a0mmol) of NH2O product gas, e.g. S1\u2010derived\u2010N2O (here: S1\u2010N2O) or S6\u2010derived\u2010N2O (S6\u2010N2O), was purified on a vacuum manifold by cryogenic distillation. Reaction by\u2010 and side\u2010products were trapped at \u221278\u00b0C (dry ice/ethanol bath); N2O was trapped at \u2212196\u00b0C (liquid N2) in a coiled stainless\u2010steel tube, while N2 and O2 (side products) were removed by evacuation with an oil\u2010sealed rotary vane pump . Thereafter, the N2O product was condensed into 10\u00a0mL stainless\u2010steel flasks under liquid\u2010nitrogen cooling. The cryogenic extraction was repeated five times to fully capture the produced N2O. Finally, the N2O yield was determined gravimetrically . The N2O purity, i.e. the absence of IR\u2010active impurities , was confirmed by FTIR spectroscopy .2 (<0.01%) and N2O recovery (>99.4%), using different gravimetric mixtures of high\u2010purity N2O and N2 .After the decomposition reaction, the N4NO3 decomposition reaction across the large range of \u03b4 values was tested. In detail, such tests were made by comparing \u03b415N\u03b1 of NH4NO3\u2010derived N2O gases (S1\u2010N2O\u2013S6\u2010N2O) with the \u03b415N(NO3\u2212) of substrate NH4NO3 salts (S1\u2013S6) and \u03b415N\u03b2 with \u03b415N(NH4+), respectively. While the link provided by the NH4NO3 decomposition reaction was assumed to be valid across a wide range of \u03b4 values, the analytics involved in \u03b415N\u03b1, \u03b415N\u03b2 or \u03b415N(NO3\u2212), \u03b415N(NH4+) analysis might display non\u2010linearities.First, the consistency of the NH2O gases S2\u2010N2O, S3\u2010N2O, S5\u2010N2O and S6\u2010N2O were analysed together with S1\u2010N2O and S4\u2010N2O using the QCLAS analyser and in preliminary experiments displayed a consistent offset between \u03b415N\u03b1(N2O), \u03b415N\u03b2(N2O) and \u03b415N(NO3\u2212), \u03b415N(NH4+) values (data not shown). For actual \u03b415N\u03b1 and \u03b415N\u03b2 of S1\u2010N2O and S4\u2010N2O, known \u03b415N(NO3\u2212) and \u03b415N(NH4+) values of the respective NH4NO3 salts were adopted and no correction for fractionation effects due to incomplete decomposition or branching isotope effects due to N2 production was applied. The uncertainty of actual \u03b415N\u03b1 and \u03b415N\u03b2 for S1\u2010N2O and S4\u2010N2O was estimated from the uncertainty of weighted mean \u03b415N(NO3\u2212) and \u03b415N(NH4+) values of the educt NH4NO3 salts S1/S4, were used to define a linear calibration function was tested against the actual \u03b415N\u03b1 values, i.e. the \u03b415N(NO3\u2212) of the respective NH4NO3 salts (S1\u2013S6). The same procedure was applied to \u03b415N\u03b2 and \u03b415N(NH4+).Measured 2.22.2.12O gases offer only limited isotopic variability. Therefore, high\u2010purity N2O was supplemented with defined amounts of 15N\u2010enriched/15N\u2010depleted and 18O\u2010enriched N2O dopant gas using a ten\u2010port two\u2010position valve with sample loops of different volumes , as well as 18O\u2010enriched N2O ((36.25\u2009\u00b1\u00a00.10)% NN16O, (63.75\u2009\u00b1\u00a00.76)% NN18O) and 15N\u03b2\u2010depleted N2O (\u03b415N\u03b1\u00a0=\u2009(\u22122.54\u2009\u00b1\u00a00.005)\u2030, \u03b415N\u03b2\u00a0=\u2009(\u2212162.21\u2009\u00b1\u00a00.03)\u2030, \u03b418O\u2009=\u2009(+38.92\u2009\u00b1\u00a00.003)\u2030), both produced and characterised at Empa. Details on the production and analysis of 18O\u2010enriched N2O and 15N\u03b2\u2010depleted N2O are provided in the supporting information\u00a0 or 150\u2009mL (Lab MPI) stainless\u2010steel flasks for isotopic analysis.The dopant gases were commercial ormation\u00a0. N2O RMs2.2.2\u03b415N\u03b1, \u03b415N\u03b2 and \u03b418O in the N2O gases, a QCLAS spectrometer \u22121 and an astigmatic Herriott multi\u2010pass absorption cell (204\u2009m path length) was applied. Prior to analysis, pure N2O gases were diluted to around 50\u2009\u03bcmol\u2009mol\u22121 using one cylinder of synthetic air ((20.5\u00a0\u00b1\u00a00.5)% O2 in N2, Messer Schweiz AG, Switzerland) into 2\u00a0L high\u2010pressure stainless\u2010steel cylinders using a ten\u2010port two\u2010position valve . A singular cylinder of synthetic air was used for all experiments to minimise differences in the oxygen content, which would otherwise affect pressure broadening of absorption lines, result in differences in apparent isotopologue mole fractions and increase uncertainties. The selection of synthetic air as diluent is somewhat arbitrary and not meant to represent an alternative for a full\u2010air matrix for high\u2010accuracy ambient N2O isotope analysis, which would enclose noble and trace gases depending on the analytics and accuracy requirements.For analysis of \u03b415N\u03b1\u00a0=\u2009(+25.73\u2009\u00b1\u00a00.24)\u2030, \u03b415N\u03b2\u00a0=\u2009(+25.44\u2009\u00b1\u00a00.36)\u2030, \u03b418O\u2009=\u2009(+35.86\u2009\u00b1\u00a00.22)\u2030) and CG2 (\u03b415N\u03b1\u00a0=\u2009(\u221248.59\u2009\u00b1\u00a00.25)\u2030, \u03b415N\u03b2\u00a0=\u2009(\u221246.11\u2009\u00b1\u00a00.43)\u2030, \u03b418O\u2009=\u2009(+27.37\u2009\u00b1\u00a00.11)\u2030). The isotopic composition of the calibration gases had been previously analysed by Sakae Toyoda at the Tokyo Institute of Technology using their analytical technique as a link to the international scales.The spectroscopically determined isotope ratios were related to the isotope\u2010ratio scales realised by Toyoda et al2O RMs by QCLAS, the site\u2010specific isotopic information provided by NH4NO3\u2010derived N2O gases S1\u2010N2O (\u03b415N\u03b1\u00a0=\u2009(\u22121.41\u2009\u00b1\u00a00.21)\u2030, \u03b415N\u03b2\u00a0=\u2009(+0.33\u2009\u00b1\u00a00.12)\u2030) and S4\u2010N2O (\u03b415N\u03b1\u00a0=\u2009(+52.36\u2009\u00b1\u00a00.15)\u2030, \u03b415N\u03b2\u00a0=\u2009(+53.06\u2009\u00b1\u00a00.16)\u2030) was propagated to the N2O RMs (RM1\u2013RM5). For this, the N2O RMs were analysed together with S1\u2010N2O and S4\u2010N2O, as described in the preceding section, to propagate the moiety\u2010specific isotopic composition defined by S1 and S4 to the novel RMs MAT 252 mass spectrometer against an isotopically characterised laboratory tank of pure N2O ; C1: \u03b415N\u2009=\u2009(\u22122.4\u00a0\u00b1\u00a00.4)\u2030, \u03b415N\u03b1\u00a0=\u2009(\u22124.5\u00a0\u00b1\u00a00.4)\u2030, \u03b415N\u03b2\u00a0=\u2009(\u22120.3\u00a0\u00b1\u00a00.8)\u2030, \u03b418O\u2009=\u2009(+23.3\u00a0\u00b1\u00a01.2)\u2030. IRMS analysis of the N2O intramolecular 15N distribution was based on the quantification of the fragment NO+ (m/z 30 and 31) and molecular N2O+ ions to calculate isotope ratios for the entire molecule and the central (\u03b1) and terminal (\u03b2) N atom. Analysis of \u03b415N (45/44) and \u03b415N\u03b1 involves correction for interfering 14N217O+ (m/z 45) and 14N17O+ (m/z 31) using actual \u039417O values analysed at the University of East Anglia (UEA). For the analysis of \u03b415N\u03b1 and \u03b415N\u03b2, rearrangement of N atoms (N\u03b1 and N\u03b2) in the ion source was considered. The \u03b415N, \u03b415N\u03b1 and \u03b415N\u03b2 values of the local reference gas were previously anchored to Air\u2010N2 by NH4NO3 decomposition,\u03b418O value was anchored to VSMOW by converting N2O into CO2 with graphite and a platinum foil . The analytical uncertainties were calculated from the uncertainty of the in\u2010house working N2O standard gases and the standard deviation for repeated measurements of the sample gas (N2O RM) and the in\u2010house working N2O standard following the law of error propagation. Specifically, the uncertainty of the in\u2010house working N2O standard gas for \u03b415N, \u03b415N\u03b1 and \u03b415N\u03b2 values comprises both the uncertainty in the \u03b415N(NH4+) and \u03b415N(NO3\u2212) analysis and the repeatability of the NH4NO3 decomposition reaction. For \u03b418O, the uncertainty of the in\u2010house working N2O standard gas includes the repeatability of the conversion reaction of N2O into CO2 with graphite. \u03b417O signatures of three N2O RMs were analysed by high\u2010resolution IRMS . Experimental details of this prototype analyses are provided in the supporting information . The system and the method used have been described by Sperlich et al.\u03b415N values of the sample N2O were scaled to IAEA\u2010N\u20101 and USGS32. In addition to the sample gases, an in\u2010house standard N2O gas NINO was analysed in each sample run, which was used as an anchor for \u03b415N measurements by DI\u2010IRMS. USGS40, and the in\u2010house standards Ali\u2010j3 (\u03b415N\u2009=\u2009(\u22121.51\u2009\u00b1\u00a00.1)\u2030; acetic anilide) and Caf\u2010j3 (\u03b415N\u2009=\u2009(\u221215.46\u2009\u00b1\u00a00.1)\u2030; caffeine), were analysed in each daily run as quality controls, but not used for data correction.2O RMs were analysed twice on a DI\u2010IRMS system using separately subsampled flasks. We note that the published \u03b4 values for USGS51 and USGS52 are average values with a rather large deviation between laboratories. Therefore, we scaled the DI\u2010IRMS \u03b415N analyses to the in\u2010house standard NINO using the value reported for the primary calibration using EA/IRMS (\u03b415N\u2009=\u2009(+0.54\u2009\u00b1\u00a00.21)\u2030). The \u03b418O value of NINO (\u03b418O\u2009=\u2009(+39.94\u2009\u00b1\u00a00.34)\u2030) was determined by setting the \u03b418O of USGS51 equal to the average value from the interlaboratory comparison (\u03b418O\u2009=\u2009(+41.45\u2009\u00b1\u00a00.34)\u2030) published by Ostrom et al.The N\u03b415N is measured from N2 gas, the DI\u2010IRMS method allows the analyses of \u03b415N and \u03b418O values by simultaneously recording m/z 44 (14N14N16O+), 45 and 46 ion currents. \u03b415N and \u03b418O values for N2O RMs were calculated according to Kaiser et al17O values determined by UEA were used.In contrast to the EA/IRMS technique, where \u03b415N and \u03b418O measurements of the N2O standard gases and from the standard deviation for repeated measurements of the sample gas (N2O RM) and the N2O standards, following the law of error propagation.The uncertainty of the analyses was calculated from the uncertainty of 2.2.52O RM samples and a N2O\u2010MG\u20106.0 working reference were diluted to 0.09\u2009mmol\u2009mol\u22121 in N2 , filled into 20\u2009mL serum vials and analysed for \u03b4(\u039d2\u039f) and \u03b4(\u039d2\u039f) on a custom\u2010built automated cryogenic extraction and purification system comprised of an autosampler, a valve system, and PoraPLOT Q pre\u2010 and main columns , coupled to a GEO 20\u201020 isotope ratio mass spectrometer .The N\u03b4(O2)\u2009=\u00a0\u03b417O,\u00a0\u03b4(O2)\u2009\u2248\u00a0\u03b418O (the error of this approximation is <0.01\u2030) and 29\u03b4(\u039d2)\u2009=\u00a0\u03b415\u039d after cryogenic N2O extraction and decomposition to N2 and O2 with a 500\u2009mm long pure gold tube held at 854\u00b0C. N2 and O2 were separated directly (without further cryofocussing) on a molecular\u2010sieve 5\u2010\u00c5 PLOT main column ). The quantitative conversion of N2O was verified by swapping the molecular\u2010sieve main column for the PoraPLOT Q main column and testing for residual N2O with the mass spectrometer. The raw \u03b417O and \u03b418O measurements were affected by scale compression. To correct for this, a logarithmic scale normalisation\u03b418O value of +112.4\u2030 (relative to N2O\u2010MG\u20106.0) derived from the \u03b4(\u039d2\u039f) measurements of the diluted RM4 sample measured on the GEO 20\u201020 mass spectrometer. The same normalisation was used for \u03b417O as for \u03b418O because no N2O reference material with a calibrated \u03b417O value was available. No scale\u2010normalisation was applied to the \u03b415N measurements. Uncertainties were calculated using the law of error propagation from the standard deviations of replicate measurements against the working reference gas and the calibration uncertainties of the working reference gas against Air\u2010N2 and VSMOW.Using the same mass spectrometer, these samples were also analysed for 2O RM samples were analysed for \u03b4(\u039d2\u039f) and \u03b4(\u039d2\u039f) with respect to the N2O working reference N2O\u2010MG\u20106.0 using the dual\u2010inlet system of a Finnigan MAT 253 isotope ratio mass spectrometer . The N2O\u2010MG\u20106.0 working reference has been calibrated by Kaiser et al,\u03b415N\u2009=\u2009(+1.01\u2009\u00b1\u00a00.06)\u2030 with respect to Air\u2010N2, as well as \u03b418O\u2009=\u2009(+38.45\u2009\u00b1\u00a00.22)\u2030 and \u03b417O\u2009=\u2009(+19.66\u2009\u00b1\u00a00.11)\u2030 with respect to VSMOW.\u03b417O values of N2O RMs analysed with the Sercon GEO 20\u201020 were used for the data correction according to Kaiser et al.2 and VSMOW.The N33.1\u03b415N\u03b1 and \u03b415N\u03b2 in N2O to the Air\u2010N2 scale and calculating uncertainties are described. Section\u00a0\u03b415N(NH4+) and \u03b415N(NO3\u2212) analyses in NH4NO3 salts (S1\u2013S6) by eight isotope laboratories against international IAEA and USGS standards. Section\u00a04NO3 decomposition at high yield, repeatability, and N2O purity. To enable the two\u2010point calibration, a number of NH4NO3 salts with different isotopic composition were produced and decomposed and the consistency of \u03b415N\u03b1 and \u03b415N\u03b2 of the N2O gases (S1\u2010N2O\u2013S6\u2010N2O) and the \u03b415N(NH4+) and \u03b415N(NO3\u2212) of NH4NO3 salts (S1\u2013S6) was tested , as analysed by the eight isotope laboratories and calibrated to Air\u2010N2 by analysis of IAEA and USGS standards, is indicated in Table\u00a04NO3 samples , all results obtained by EA/IRMS were included for calculation of the weighted mean value except for results by one laboratory (Lab 4), as this laboratory used a more complicated analytical procedure with higher uncertainties. For \u03b415N(NO3\u2212) and \u03b415N(NH4+), all laboratory results were included to calculate the weighted mean values, irrespective of the applied analytical technique.For \u03b415N(NH4NO3) with average \u03b415N(NH4+)/\u03b415N(NO3\u2212) values indicates a good agreement to within <0.2\u2030. Nonetheless, results by individual laboratories for moiety\u2010specific \u03b4 values deviate substantially from the weighted mean. As an example, \u03b415N(NO3\u2212) results from Lab 8 are substantially higher than those from other laboratories by an average of (+2.15\u2009\u00b1\u00a00.58)\u2030. This may be due to the specific preparation technique applied, NH4+ removal by ion exchange, a technique which is prone to preferential retention/elution of 15NO3.\u03b415N values of both NO3\u2212 and NH4+, which may be reflected in the \u03b415N(NH4+) values of Lab 6 but not those of Lab 5, where a similar technique was used. Conversely, systematic fractionation effects by preparation techniques should be accounted for by identical treatment (IT) of the provided IAEA and USGS standards used for data correction. In summary, analysis of \u03b415N(NH4+) and \u03b415N(NO3\u2212) is still challenging; however, the ensemble of techniques applied in this study provides good agreement with \u03b415N(NH4NO3) values.A comparison of 3.1.22O yield of 93\u201395% was achieved for the decomposition of NH4NO3 salts S1\u2013S6 (Table\u00a0n\u00a0=\u00a03), surpasses results by Westley et aln\u00a0=\u200920). A further increase in the yield of the NH4NO3 decomposition was achieved by conducting the reaction in a NH4HSO4\u2013(NH4)2SO4 melt (around 2%), as suggested for industrial applications by Szab\u00f3 et al\u03b415N\u03b1 values but a loss in the \u03b415N\u03b2 information due to NH4+ salt addition, and was thus not continued.Under optimised reaction conditions and distillation procedure, an average N\u03b415N\u03b1 and \u03b415N\u03b2 for the loss in N2O (around 5\u20137%), mainly due to uncertainties in the reaction mechanisms (incomplete decomposition or side\u2010reaction), which makes it difficult to estimate the effect on \u03b4 values. Assuming incomplete reaction accompanied by fractionation effects, according to our earlier study,\u03b415N\u03b1/\u03b415N\u03b2/\u03b415N values, respectively. However, a much smaller difference in \u03b415N was observed when comparing results of N2O RMs analysed by QCLAS with IRMS analyses. Therefore, our assumption is that the decrease in yield is at least partly caused by a \u201cbranching\u201d side reaction, e.g. nitrogen gas (N2) production,\u03b415N(N2) values.2 production has a minor effect on \u03b415N\u03b1, \u03b415N\u03b2 and \u03b415NSP, but the effect is expected to depend on the timing of N2 generation, which is not known.No correction was applied to 3.1.32 scale and to determine the N2O site\u2010specific isotopic composition across a wide range of \u03b4 values. Therefore, the consistency of the isotopic composition of the N2O gases and the NH4NO3 salts (\u03b415N(NH4+) and \u03b415N(NO3\u2212), S1\u2013S6) was tested. The detailed procedure is described in section\u00a04NO3 decomposition reaction, measured \u03b415N\u03b1 values of S1\u2010N2O/S4\u2010N2O and actual \u03b415N\u03b1 values, i.e. \u03b415N(NO3\u2212) of the educt NH4NO3 salts S1/S4, were used to define a linear calibration function. \u03b415N\u03b1cal values of S2\u2010N2O, S3\u2010N2O, S5\u2010N2O and S6\u2010N2O were calculated from measured \u03b415N\u03b1 using this correction function and compared against actual values /\u03b415N(NH4+)/\u03b415N(NO3\u2212) \u2013 \u03b415N(NH4+) of the respective salts . Assuming similar fractionation effects for decomposition of all NH4NO3 salts (S1\u2013S6), provided the comparable decomposition yield and \u03b415N(NH4+) analyses of the NH4NO3 salts. The latter is more plausible, as the QCLAS analyses using the same calibration approach showed good agreement with independent IRMS measurements for N2O RM with high 15N enrichment (e.g. S5), which agrees with earlier studies indicating challenges in \u03b415N(NH4+) analysis, but this may also be due to the lack of available international standards for \u03b415N(NH4+) that cover \u03b4 values above (+53.75\u2009\u00b1\u00a00.24)\u2030 (USGS26) and below (\u221230.41\u2009\u00b1\u00a00.27)\u2030 (USGS25).Results of O3 salts \u2013S6, prov2O gases from around zero (S1\u2010N2O) to 15N\u2010enriched (S4\u2010N2O) and of the substrate NH4NO3 salts (S1\u2013S4). Thereby, our study covers a much larger range of \u03b4 values than earlier studies,2 scale. At very high and low 15N enrichment , the calibration approach using NH4NO3 decomposition is more challenging, probably due to less satisfying analytical accuracy of \u03b415N(NH4+) measurements to date. As the N2O gases S5\u2010N2O and S6\u2010N2O were not included in the analysis of N2O RMs, their enhanced uncertainty in \u03b415N\u03b1cal and \u03b415N\u03b2cal does not affect the data quality of N2O RMs.In summary, our results demonstrate consistency of the isotopic composition of the N3.23.2.12O RMs (RM1\u2013RM5) were calibrated against Air\u2010N2 by both QCLAS (Lab Empa) and IRMS (Lab TT) analyses. For QCLAS analyses, two N2O gases produced by NH4NO3 decomposition were applied to define a calibration function and propagate the isotopic information of the NH4NO3 salts (\u03b415N(NO3\u2212), \u03b415N(NH4+)) to the N2O RMs . \u03b415NSP and \u03b415N were calculated using definitions and their uncertainty estimated using the law of error propagation. In Table\u00a0\u03b415NSP values acquired by QCLAS (Lab Empa) using the calibration approach established in this study are compared with results provided by DI\u2010IRMS (Lab TT) using a previously published link to the Air\u2010N2 scale.2O RMs are shown in Tables\u00a0The novel N\u03b415NSP measurements by DI\u2010IRMS and QCLAS (Lab Empa) across all N2O RMs. This is most likely attributable to the calibration of the position\u2010dependent \u03b4 values with respect to Air\u2010N2 via the NH4NO3 decomposition technique, which were performed independently for the two labs. Incidentally, for the NH4NO3 salts S1\u2013S4, the \u03b415N(NO3\u2212) results provided by Lab TT were always lower ((\u22120.63\u2009\u00b1\u00a00.59)\u2030), while \u03b415N(NH4+) values were higher than the respective weighted mean values ((+0.49\u2009\u00b1\u00a00.25)\u2030), which would lead to 1.12\u2030 lower \u03b415NSP values \u2030 vs. (+18.7\u00a0\u00b1\u00a02.2)\u2030 for ambient tropospheric N2O.\u03b415NSP measurements by the Tokyo Institute of Technology, using the NH4NO3 decomposition technique.A similar 1.5\u20132.0\u2030 difference in \u03b415NSP and the Air\u2010N2 scale with high accuracy is still challenging and the current realisation of the Air\u2010N2 scale for \u03b415NSP provided by USGS51 and USGS52\u03b415NSP values and should be revisited in future studies.We conclude that the realisation of the link between 3.2.2\u03b415N values of N2O RMs were analysed by IRMS in three different laboratories using independent links to the Air\u2010N2 scale \u2030 (USGS51) and (+0.07\u2009\u00b1\u00a00.22)\u2030 (USGS52). These values fall between results published by laboratories 7 and 8 (USGS and BGC\u2010IsoLab) in Ostrom et al,\u03b415N(N2O) measurements. A similar offset between laboratories had already been detected earlier and was attributed to differences in the propagation of the Air\u2010N2 scale to \u03b415N(N2O).2O RMs.le Table\u00a0. Results52 Table\u00a0 by a com17O data are available. Consistency of N2O RM flask subsamples was demonstrated using DI\u2010IRMS and 0.03\u2030 for \u03b418O(N2O) .In contrast, differences between analytical techniques applied within one lab, thus using the same link to the scale, were smaller than offsets between laboratories: (+0.10\u00a0\u00b1\u00a00.04)\u2030 for Lab MPI and (+0.12\u2009\u00b1\u00a00.14)\u2030 for Lab UEA. This indicates that both EA/IRMS and GC\u2010IRMS and DI\u2010IRMS can achieve high accuracy, provided that an accurate link to the scale and \u03943.2.3\u03b418O values of N2O RMs were analysed by IRMS in three different laboratories , indicating a potential scaling or scale compression issue. Measurements were not completely independent for all laboratories, as the results for Lab MPI\u2010II were referenced to average \u03b418O values of USGS51 in Ostrom et al,es Table\u00a0. Results\u03b417O values were determined by GC/IRMS at UEA (Lab UEA\u2010I) and showed a (0.98\u2009\u00b1\u00a00.27)\u2030 offset to prototypical measurements by HR\u2010IRMS (MAT253 ULTRA) at the Tokyo Institute of Technology 2SO4 melt. n indicates the number of decomposition experiments.Table S2. Isotopic composition of RMs, analysed by QCLAS at Empa versus S1\u2010N2O and S4\u2010N2O to calculate \u03b415N\u03b1, \u03b415N\u03b2, \u03b415NSP, and \u03b415N values. n indicates the number of analyses. Uncertainties are calculated using the law of error propagation involving the uncertainties in S1\u2010N2O and S4\u2010N2O, their analyses, and the analyses of N2O RMs but do not enclose deviations due to fractionation or branching effects during NH4NO3 decomposition. All values are reported in \u2030.Table S3. Isotopic composition of RMs, analysed by IRMS at Tokyo Institute of Technology (Lab TT) versus an isotopically characterised in\u2010house working standard to calculate \u03b415N\u03b1, \u03b415N\u03b2, \u03b415NSP, \u03b415N, and \u03b418O values on the \u201cTokyo Tech scale\u201d. n indicates the number of analyses. Uncertainties are calculated using the law of error propagation. All values are reported in \u2030.Table S4.\u03b415N of RMs, the in\u2010house N2O standard gas (NINO), and a number of quality control standards, analysed by Lab MPI versus primary reference materials and second scale anchor of the Air\u2010N2 scale . n indicates the number of analyses. Expanded uncertainties are calculated following the law of error propagation. For the quality control, standards target values and references are provided as well. All values are reported in \u2030.Table S5. Isotopic composition of RMs, analysed as N2O diluted to 0.09\u2009mmol\u2009mol\u22121 on Sercon GEO 20\u201320 IRMS (UEA\u2010I) after gold decomposition, scale\u2010normalised to the \u03b418O value of RM4. n indicates the number of analyses. Uncertainties are calculated using the law of error propagation from the standard deviations of replicate measurements against the working reference gas and the calibration uncertainties of the working reference gas against Air\u2010N2 and VSMOW.14 All values are reported in \u2030.Table S6. Isotopic composition of RMs, analysed as pure N2O on Finnigan MAT 253 IRMS (UEA\u2010II) using the actual \u039417O measurements. n indicates the number of analyses. Uncertainties are calculated using the law of error propagation from the standard deviations of replicate measurements against the working reference gas and the calibration uncertainties of the working reference gas against Air\u2010N2 and VSMOW.15 All values are reported in \u2030.Click here for additional data file."} +{"text": "The main difference between mol\u00adecules A and B involves the dihedral angles of the phenyl groups. One phenyl ring makes dihedral angles of 71.14\u2005(6)\u00b0 (mol\u00adecule A) and 82.81\u2005(7)\u00b0 (mol\u00adecule B) with respect to the core (C4N3O2) of the mol\u00adecule [14.56\u2005(9)\u00b0 (mol\u00adecule A) and 5.7\u2005(1)\u00b0 (mol\u00adecule B) for the other phenyl ring]. Another prominent feature is the intra\u00admolecular N\u2014H\u22efO hydrogen bond present in both crystallographically independent mol\u00adecules.The title compound, C E)-N-Phenyl-N-(phenyl\u00adcarbamo\u00adyl)-3-[prop\u00adyl(tri\u00admethyl\u00adsil\u00adyl)amino]\u00adacryl\u00adamide is an insertion product from prop\u00adyl(tri\u00admethyl\u00adsil\u00adyl)[2-(tri\u00admethyl\u00adsil\u00adyl)ethen\u00adyl]amine and phenyl iso\u00adcyanate. It was obtained in the course of our work on different types of silicon\u2013nitro\u00adgen compounds carbamo\u00adyl]-3-(meth\u00adyl\u00adsulfan\u00adyl)acryl\u00adamide \u2005\u00c5 in mol\u00adecule B] The planarity is presumably due to the conjugated system of double bonds. The C14\u2013C19 phenyl rings in both mol\u00adecules are not coplanar to the core of the mol\u00adecules but adopt dihedral angles to the latter of 14.56\u2005(9)\u00b0 (mol\u00adecule A) and 5.7\u2005(1)\u00b0 (mol\u00adecule B). This small deviation from planarity still allows conjugation between the C14\u2013C19 phenyl ring and the urea part of the mol\u00adecule.The core of the mol\u00adecule formed by N1, C4\u2013C6, O1, N2, C7, O2 and N3 is almost planar in both mol\u00adecules of the title compound [the average deviation from the plane is 0.05\u2005(6)\u2005\u00c5 in mol\u00adecule A) and 82.81\u2005(7)\u00b0 (mol\u00adecule B) with the core of the mol\u00adecule. This almost perpendicular conformation may be explained by the presence of the oxygen atom O2 in a vicinal position to the respective phenyl group.The C8\u2013C13 phenyl rings in both mol\u00adecules subtend dihedral angles of 71.14\u2005(6)\u00b0 -N-phenyl-N-(phenyl\u00adcarbamo\u00adyl)-3-[prop\u00adyl(tri\u00admethyl\u00adsil\u00adyl)amino])acryl\u00adamide was obtained from the reaction of prop\u00adyl(tri\u00admethyl\u00adsil\u00adyl)[2\u2013(tri\u00admethyl\u00adsil\u00adyl)ethen\u00adyl]amine and phenyl iso\u00adcyanate. As shown in Fig.\u00a04et al., 2018(n-pentane were added dropwise 0.35\u2005g (3\u2005mmol) phenyl\u00adiso\u00adcyanate at 0\u00b0C. After standing for six days at room temperature, volatiles were removed under reduced pressure. Storing the product mixture for five years at \u221228\u00b0C yielded crystals suitable for single-crystal X-ray diffraction. No qu\u00adanti\u00adtative yield can be given here, since only a few crystals at the wall of the Schlenk tube were available. NMR spectroscopy showed that the batch product is a mixture of many components. Further purification of the product mixture was not successful.To a solution of 0.46\u2005g (2\u2005mmol) prop\u00adyl(tri\u00admethyl\u00adsil\u00adyl)[2-(tri\u00admethyl\u00adsil\u00adyl)ethen\u00adyl]amine in 10\u2005ml Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314623001177/im4017sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314623001177/im4017Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314623001177/im4017Isup3.cmlSupporting information file. DOI: 2240673CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of the title compound has been characterized by single-crystal X-ray diffraction and Hirshfeld surface analyses. The mol\u00adecular structure and frontier orbitals were also investigated using DFT. 11H9BrN2O, is almost planar. The benzene and pyrimidine rings are essentially coplanar, with r.m.s. deviations of 0.0130\u2005\u00c5, and the largest displacement is for the flap atom of the di\u00adhydro\u00adpyrrole moiety [0.154\u2005(7)\u2005\u00c5]. Hirshfeld surface analyses revealed that the crystal packing is dominated by H\u22efH, Br\u22efH/H\u22efBr and O\u22efH/H\u22efO inter\u00adactions, and Br\u22efBr inter\u00adactions in the crystal structure are also observed. Theoretical calculations using density functional theory (DFT) with the B3LYP functional basis set gave numerical parameters for the frontier molecular orbitals.The mol\u00adecular structure of the title compound, C In comparison with a reported literature procedure at room temperature (293\u2013298\u2005K) and the reaction products separated by extraction after the reaction mixture was reduced to pH = 9\u201310 with NH4OH. As distinguished from the reported procedure, we carried out these reactions by cooling in an ice bath at a much lower temperature (273\u2013275\u2005K) and for a relatively longer period of time. The reaction products were finally separated by cold NH4OH at pH = 10\u201311. In general, the inter\u00adactions of 7-bromo-2,3-di\u00adhydro\u00adpyrrolo\u00adquinazolin-9(1H)-one with aldehydes are well-studied with the B3LYP functional basis set.2.et al., 2009et al., 2007aet al., 1992et al., 2009et al., 2005The mol\u00adecular structure of the title compound is shown in Fig.\u00a013.et al., 1964Cg inter\u00adactions, with Br\u22efCg1 = 3.6428\u2005(15)\u2005\u00c5, forming a layered network \u2005\u00c5 and a ring slippage of 1.569\u2005\u00c5, and Cg2\u22efCg3ii being 3.7513\u2005(16)\u2005\u00c5 and a ring slippage of 1.194\u2005\u00c5. Both short inter\u00admolecular contacts help to stack parallel mol\u00adecules along [100]. The resulting two-dimensional network extends parallel to (002), with neighbouring layers linked through C1\u2014H1B\u22efN4 short inter\u00admolecular contacts, H1B\u22efN4 = 2.73\u2005\u00c5, C1\u2014H1B\u22efN4 = 169\u00b0, to form the full three-dimensional structure \u2005\u00c5, which is shorter than the sum of van der Waals radii to the Hirshfeld surface . Besides these contacts, Br\u22efH/H\u22efBr (19.6%), O\u22efH/H\u22efO (11.3%), N\u22efH/H\u22efN (8.1%) and C\u22efH/H\u22efC (6.9%) inter\u00adactions contribute significantly to the total Hirshfeld surface; their decomposed fingerprint plots are shown in Fig.\u00a06c\u2013f. The contributions of further contacts are only minor and amount to N\u22efC/C\u22efN (3.5%), O\u22efC/C\u22efO (2.0%), Br\u22efC/C\u22efBr (0.9%), Br\u22efBr (0.8%), O\u22efN/N\u22efO (0.5%) and Br\u22efN/N\u22efBr (0.3%).In order to qu\u00adantify the inter\u00admolecular inter\u00adactions in the crystal of the title compound, a Hirshfeld surface (HS) analysis , chemical hardness (h) and chemical softness (s) can be calculated as follows: c = (I + A)/2; h = (I - A)/2; s = 1/2h, where I and A are the ionization potential and electron affinity, respectively, where I = \u2013EHOMO and A = \u2013ELUMO , hardness (h), potential (m), electrophilicity (w) and softness (s) for the title mol\u00adecule were calculated at the DFT/B3LYP level using the 6-311++G basis set indicates the relatively high stability of the title mol\u00adecule.t Table\u00a01. The val6.et al., 2016b]quinazolin-9(1H)-one moiety with a similar conformation to that in the title structure: de\u00adoxy\u00advasicinone -one moiety gave only two hits: N-propanamide sesquihydrate -one mono\u00adhydrate (0.142\u2005mol) of phospho\u00adroxychloride were added dropwise over 1\u2005h at 273\u2013275\u2005K. The reaction mixture was then heated at 368\u2013371\u2005K for 2\u2005h, it was subsequently cooled and finally poured over ice. The temperature of the mixture was kept at around 273\u2013275\u2005K. When the reaction mixture was completely decomposed, it was brought to pH = 10\u201311 with 25%wt ammonium hydroxide solution. The light-yellow precipitate was filtered off, dried and recrystallized from methanol. The yield of the product was 4.35\u2005g (82%), m.p. 431\u2013433\u2005K : 8.4 , 7.8 , 7.5 , 4.2 , 3.18 , 2.31 .8.Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022007800/wm5655sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022007800/wm5655Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022007800/wm5655Isup3.cmlSupporting information file. DOI: 2194365CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal of the title flavone derivative, O\u22efI halogen bonding and T\u2013shaped \u03c0\u2013stacking combine to generate a herringbone packing motif. 19H13IO2, the dihedral angle between the naphthyl ring system and the pendant iodo\u00adphenyl ring is 72.48\u2005(11)\u00b0. In the crystal, C\u2014H\u22ef\u03c0 inter\u00adactions and I\u22efO [3.293\u2005(2)\u2005\u00c5] halogen bonds are observed, which combine to generate a herringbone packing motif.In the title compound, C Our group is particularly inter\u00adested in the extended flavonol motif as it has been shown to release CO qu\u00adanti\u00adtatively with visible light (Popova Cg1\u22efCg2i = 4.929\u2005(2)\u2005\u00c5 and C6\u2014H6\u22efCg2i = 154.5\u2005(3)\u00b0, where Cg1 is the centroid of the pyran\u00adone ring containing atoms C4\u2013C7/C12/C13 and Cg2 is the centroid of the iodo\u00adphenyl ring containing atoms C14\u2013C19 \u2005\u00c5, with C17\u2014I1\u22efO2ii and I1\u22efO2ii\u2014C1ii angles of 177.21\u2005(10) and 127.9\u2005(2)\u00b0, respectively . This I\u22efO separation is some 0.25\u2005\u00c5 shorter than van der Waals\u2019inter\u00adaction distance of 3.5\u2005\u00c5 was added and the reaction was stirred until a precipitate formed. The reaction mixture was acidified to pH 4 with glacial acetic acid. The solids were filtered and taken directly to the next step. (E)-1--3-(4-iodo\u00adphen\u00adyl)prop-2-en-1-one was then suspended in ethanol (10\u2005ml). An NaOH solution was added and the reaction stirred until a precipitate formed. The reaction mixture was acidified to pH 1 with HCl (6 M). The white solid was collected by filtration and slow evaporation of a solution of the title compound in ethyl acetate gave colorless crystals .1-Acetyl-2-naphthol and 4-iodo\u00adbenzaldehyde were dissolved in ethanol (5\u2005ml). An NaOH solution \u03b4 = 9.46 , 7.95 , 7.80\u20137.75 , 7.65 , 7.44 , 7.26 , 7.16 , 5.54 , 3.16 , 2.95 ppm.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2414314620001108/hb4334sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314620001108/hb4334Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314620001108/hb4334Isup3.cmlSupporting information file. DOI: 1980111CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The TaV atom has a distorted tetra\u00adhedral coordination environment in a three-legged piano-stool fashion. The conformation of the penta\u00adfulvene exocyclic C atom to the three other ligands is staggered and not eclipsed, as found in the crystal structure of 1. Inter\u00admolecular inter\u00adactions include \u03c0\u2013\u03c0 stacking, H\u22ef\u03c0 inter\u00adactions and weak C\u2014H\u22efCl hydrogen bonds.The reaction of (\u03b7 The C6exo atom coordinates roughly opposite of Cl2 to the central tantalum atom [C6\u2014Ta1\u2014Cl2: 171.58\u2005(3)\u00b0]. Relative to the centroid of the five-membered ring (Ct), the angles to the chloride ligands are smaller than to the nitro\u00adgen ligands . The bond length Ta1\u2014N1 [2.0433\u2005(9)\u2005\u00c5] and the sum of angles at N1 [347.1\u2005(2)\u00b0] indicates a weak inter\u00adaction of the nitro\u00adgen lone pair with the metal. The penta\u00adfulvene coordinates in a \u03c0-\u03b75:\u03c3-\u03b71 fashion and exhibits typical distortion parameters . The C\u2014C bond lengths within the penta\u00adfulvene are summarized in Fig.\u00a02b. The penta\u00adfulvene has a ring slippage \u0394 of 0.31\u2005\u00c5 and a \u03b8 angle of the Cipso\u2014Cexo bond out of the plane of the five-membered ring of 36.30\u2005(12)\u00b0. The Cipso\u2014Cexo bond is a single to double bond . Consequently, the Ta1\u2014Cl2 bond [2.3965\u2005(3)\u2005\u00c5) is longer than the Ta1\u2014Cl1 bond [2.3452\u2005(3)\u2005\u00c5]. These pairs form a double-chain , linked by supra\u00admolecular contacts of the penta\u00adfulvene and the carbazolide ligands via \u03c0\u2013\u03c0 stacking [C1\u22efC17: 3.3867\u2005(15)\u2005\u00c5] and an H\u22ef\u03c0 inter\u00adaction [C15\u22efH10c: 2.773\u2005(6)\u2005\u00c5]. Numerical details of other hydrogen-bonding inter\u00adactions are summerized in Table\u00a01On the supra\u00admolecular level, around an inversion center, two mol\u00adecules mutually inter\u00adact in Fig.\u00a03b, linke1 was prepared according to Riley et al. was dissolved in tetra\u00adhydro\u00adfuran (20\u2005ml) and cooled to 223\u2005K. One equivalent of etheric HCl was added dropwise and the solution was slowly brought to room temperature. After stirring over night, the solvents were removed in vacuo and the residue was extracted with toluene (10\u2005ml). The solution was diluted with n-hexane (10\u2005ml) and stored at 277\u2005K for three days to yield a red crystalline material containing 1 and 2 (1:1). 1H NMR : \u03b4 = 0.79 , 0.85 , 1.27 , 1.49 , 1.53 , 2.13 , 2.62 , 3.27 , 3.65 , 6.30\u20138.29 p.p.m.Complex SHELXL . Refinement with OLEX2 . Refining all atoms anharmonically was dismissed, because it lowers the reliability factors only marginally, but more than triples the refinement parameters (263 versus 888 parameters).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314622012019/wm4176sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622012019/wm4176Isup2.hklStructure factors: contains datablock(s) I. DOI: 2231842CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Cp rings in both ferrocene groupings are close to eclipsed. In the crystal, O\u2014H\u22efO hydrogen bonds link the mol\u00adecules into [100] chains.In the title compound, [CoFe The resulting complexes may have applications in the fields of electroluminescent devices, environmental sensors, photodynamic therapy and biological imaging \u00b0. The Cp rings of both ferrocene groups are in nearly eclipsed conformations.The cobalt(II) atom has a octa\u00adhedral coordination environment defined by two tri\u00adfluoro\u00admethyl-\u03b2-diketone ferrocene ligands and two water ligands Fig.\u00a01 with thex, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z) contact .In the crystal, the mol\u00adecules are linked into [100] chains by O\u2014H\u22efO hydrogen bonds arising from the water mol\u00adecules Table\u00a01; an intrIn a 250\u2005ml round-bottom flask, tri\u00adfluoro\u00admethyl-\u03b2-diketone ferrocene (0.52\u2005g 1.6\u2005mmol), tri\u00adethyl\u00adamie (0.25\u2005g 2.45\u2005mmol) and cobalt acetate (0.13\u2005g 0.5\u2005mmol) were dissolved in 100\u2005ml of methanol and the mixture was stirred at 343\u2005K for 12\u2005h and then cooled to room temperature. A red solid was obtained by suction filtration. Crystals for X-ray analysis were obtained by recrystallization from methanol solution.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314620012407/hb4360sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314620012407/hb4360Isup3.hklStructure factors: contains datablock(s) I. DOI: 2031062CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A series of O\u2014H\u22efN inter\u00adactions connect the mol\u00adecules into twisting wires, which are cross-linked through van der Waals inter\u00adactions.A co-crystal consisting of a 1:1 ratio of p-vanillic acid (1/1)], C12H10N2\u00b7C8H8O4, the dihedral angle between the pyridine rings is 59.51\u2005(5)\u00b0. In the crystal, the mol\u00adecules are linked by O\u2014H\u22efN hydrogen bonds, generating [401] chains of alternating C12H10N2 and C8H8O4 mol\u00adecules.In the title 1:1 co-crystal [alternatively called bi\u00adpyridine ethyl\u00adene\u2013 The vanillic acid has two distinct O\u2014H\u22efN-type hydrogen-bonding inter\u00adactions (Table\u00a01para-position hydroxyl group and the other pyridine N atom of a BPyE mol\u00adecule resulting in a 2.6868\u2005(13)\u2005\u00c5 distance between heteroatoms was added to a 25\u2005ml scintillation vial to which methanol was added until both compounds dissolved (approximately 20\u2005ml). The resulting solution was vortexed for 30\u2005s at 3000 rpm on a VWR Mini Vortexer MV I. The solution was then stored in the dark uncapped to allow for crystal formation while the solvent slowly evaporated.A 1:1 molar ratio of bi\u00adpyridine ethyl\u00adene and Crystal data, data collection, and structure refinement details are summarized in Table\u00a0210.1107/S2414314622003042/hb4402sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622003042/hb4402Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622003042/hb4402Isup3.cmlSupporting information file. DOI: 2160226CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The structure exhibits disorder of one of the 4-meth\u00adoxy\u00adbenzyl groups, the hy\u00addroxy group bonded to the 1,3-diazinan ring, and the methyl group of the meth\u00adoxy residue. The crystal packing is sustained by C\u2014H\u22efO and O\u2014H\u22ef\u03c0 inter\u00adactions, giving rise to infinite chains running along the b-axis direction.The title compound resulted from the condensation reaction between 1,3-bis\u00ad{[(4-meth\u00adoxy\u00adphen\u00adyl)meth\u00adyl]amino}\u00adpropan-2-ol and 2-hy\u00addroxy\u00adbenzaldehyde in CH 26H30N2O4, at 173\u2005K has ortho\u00adrhom\u00adbic (Pbca) symmetry. It was previously described by Bolte et al. . The title compound resulted from the condensation reaction between 1,3-bis\u00ad{[(4-meth\u00adoxy\u00adphen\u00adyl)meth\u00adyl]amino}\u00adpropan-2-ol and 2-hy\u00addroxy\u00adbenzaldehyde in CH3OH. The structure exhibits disorder. One of the 4-meth\u00adoxy\u00adbenzyl groups, the hy\u00addroxy group bonded to the 1,3-diazinan ring, and the methyl group of the meth\u00adoxy residue are disordered over two orientations, with occupancies of 0.807\u2005(3)/0.193\u2005(3), 0.642\u2005(5)/0.358\u2005(5), and 0.82\u2005(4)/0.18\u2005(4), respectively. The dihedral angles between the mean planes of the central 1,3-diazinan-5-ol and the 4-meth\u00adoxy\u00adphenyl rings (both occupancy components of the disordered ring) are 88.65\u2005(13), 85.79\u2005(14) and 83.4\u2005(7)\u00b0. The crystal packing is sustained by C\u2014H\u22efO and O\u2014H\u22ef\u03c0 inter\u00adactions, giving rise to infinite chains running along the b-axis direction.The redetermined structure of 2-(2-hy\u00addroxy\u00adphen\u00adyl)-1,3-bis\u00ad(4-meth\u00adoxy\u00adbenz\u00adyl)-1,3-diazinan-5-ol, C It is noteworthy that the coupling constants with magnitudes between 2.9 and 3.1\u2005Hz provide a strong evidence of the presence of an axial OH group. In this regard, it has been reported that the presence of an intra\u00admolecular hydrogen bond may stabilize the hydroxyl group in the otherwise non-preferred axial position meth\u00adyl]amino}\u00adpropan-2-ol, easily obtained following the reported method \u2005\u00c5, \u03b8 = 3.1\u2005(3)\u00b0, \u03c6 = 250\u2005(4)\u00b0, Q(2) = 0.035\u2005(3)\u2005\u00c5 and Q(3) = \u22120.561\u2005(3)\u2005\u00c5. Atoms C2 and C3 deviate from the mean plane of the other four atoms by \u22120.242\u2005(3) and 0.249\u2005(3)\u2005\u00c5, respectively. Atoms N1 and N2 are essentially tetra\u00adhedral (bond-angle sums are 331.8\u00b0 for N1 and 330.1\u00b0 for N2), while the usual \u03a3H\u2013N\u2013H angle in an ammonia mol\u00adecule is 321\u00b0 \u2005\u00c5] and the C21\u2013C26, C31\u2013C36 and C31\u2032\u2013C36\u2032 phenyl rings of the 1,3- benzyl groups are 88.65\u2005(13), 85.79\u2005(19) and 83.4\u2005(7)\u00b0, respectively, whereas the mean plane is rotated by 81.22\u2005(13)\u00b0 towards the C11\u2013C16 phenyl ring of the 2-hy\u00addroxy\u00adphenyl substituent. The dihedral angles between this phenyl ring and the other two phenyl rings are 55.54\u2005(13)\u00b0 (C21\u2013C26), 84.27\u2005(19)\u00b0 (C31\u2013C36) and 77.9\u2005(7)\u00b0 (C31\u2032\u2013C36\u2032), respectively.S(6) graph-set motif (Table1). The N\u22efO distance [2.740\u2005(3)\u2005\u00c5] is long in comparison with the values observed in related structures is shorter than 4\u2005\u00c5. It is noteworthy that the inter\u00adplanar distance between the symmetry-related main parts of the C31\u2013C36 ring is only 3.66\u2005\u00c5; however, the corresponding Cg\u22efCg distance is too long at 4.619\u2005(3)\u2005\u00c5, indicating a significant horizontal shift of the rings precluding \u03c0\u2013\u03c0 stacking.In the crystal structure, the mol\u00adecules are inter\u00adlinked ds Fig.\u00a03 into chas Table\u00a01 amino}\u00adpropan-2-ol in methanol (20\u2005mL) salicyl\u00adaldehyde was added dropwise. The resulting mixture was heated at reflux for 2\u2005h and allowed to cool to room temperature. The solvent was removed under vacuum and the crude solid was washed with cold methanol and dried 1H NMR \u03b4 7.15 , 7.12\u20137.13 , 7.11 , 6.99 , 6.85 , 6.81 , 3.88 , 3.76 , 3.72\u20133.74 , 3.03 , 2.99 , 2.98 , 2.22 , 1.60 . The hydrogen atom of a hydroxyl group could not be assigned because of the overlaping and widening of that signal with those due to hydrogen bonds.26H30N2O4. Calculated, %: C, 71.89; H, 6.91; N, 6.45; O, 14.75.Elemental analysis : Found, %: C 71.87; H 6.91; N 6.45; O 14.74. C6.Uiso values of methyl H atoms were set to 1.5Ueq(C), while the Uiso values of H atoms bonded to the remaining C atoms were set to 1.2Ueq(C). The H atom bonded to O in the major occupied site was freely refined. The H atom bonded to O in the minor occupied site was refined using a riding model with Uiso(H) set to 1.5Ueq(O). In addition, the H\u2014O\u2014C\u2014C torsion angle was allowed to refine. The displacement ellipsoids of O4 and O4\u2032 were restrained to be similar. The distances O4\u2014C3 and O4\u2032\u2014C3 were restrained to be similar. Bond lengths and angles in the fragments C24\u2013O2\u2013C27\u2032 and C24\u2013O2\u2013C27 were restrained to be similar. The displacement ellipsoids of O2 and C27/C27\u2032 were restrained to be similar. Bond lengths, angles and displacement parameters in the fragments N2\u2013O3\u2032\u2013C31\u2032\u2013C32\u2032\u2013C33\u2032\u2013C34\u2032\u2013C35\u2032\u2013C36\u2032\u2013C37\u2032\u2013C8\u2032 and N2\u2013O3\u2013C31\u2013C32\u2013C33\u2013C34\u2013C35\u2013C36\u2013C37\u2013C8 were restrained to be similar. The following restraints implemented in SHELXL were used to restrain the geometry and ijU of the disordered parts.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022006508/jq2018sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022006508/jq2018Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022006508/jq2018Isup3.cmlSupporting information file. DOI: 2092230CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II part of aqua\u00adzinc(II) diperchlorate, [Zn(C8H20N4)H2O](ClO4)2, exhibits a slightly distorted square-pyramidal coordination environment with a water mol\u00adecule in the apical position.The cationic Zn II part of aqua\u00adzinc(II) bis\u00ad(perchlorate), [Zn(C8H20N4)(H2O)](ClO4)2, exhibits a slightly distorted square-pyramidal coordination environment with a water mol\u00adecule in the apical position. In the crystal, the macrocyclic ring alternates between two conformations with equal occupancies. Two of the three perchlorate anions are situated about a twofold rotation axis, and one of them shows disorder of the O atoms with occupancies of 0.62\u2005(7) and 0.38\u2005(7). In the crystal, the complexes are connected by inter\u00admolecular hydrogen bonding via the perchlorate anions.The cationic Zn The macrocyclic ring is disordered, and two alternate conformations of each N\u2013C\u2013C\u2013N bridge can be observed (conformation A and B) in the basal plane, with a ZnII-bound H2O mol\u00adecule occupying the apical position. Addison et al. (1984\u03b2\u00a0\u2212\u00a0\u03b1)/60\u00b0] to determine if the five-coordinate atom has a square-pyramidal or trigonal\u2013pyramidal coordination environment. The bond angles \u03b2 and \u03b1 are the largest and second-largest in the coordination sphere, respectively; an ideal square pyramid and an ideal trigonal bipyramid have \u03c4 = 0 and 1, respectively. In conformation A, the N\u2014ZnII\u2014N bond angles \u03b1 and \u03b2 are 138.2\u2005(3)\u00b0 and 138.7\u2005(3)\u00b0, respectively; the corresponding bond angles in conformation B are 137.4\u2005(4)\u00b0 and138.7(4)\u00b0. The \u03c4 values are 0.008 and 0.022 for conformations A and B, respectively. Therefore, the coordination geometry around the central ZnII cation can be described as slightly distorted square-pyramidal. The occupancies for the non-hydrogen atoms of cyclen except for the four carbon atoms were set to 0.50. Atom Zn1 is 0.755\u2005(5) and 0.763\u2005(3)\u2005\u00c5 above the basal plane formed by four N atoms in conformations A and B, respectively. The Zn1\u2014O1 bond length [1.9721\u2005(4)\u2005\u00c5] is within the typical range [1.94\u20132.03\u2005\u00c5] for similar five-coordinated Zn complexes 2 Fig.\u00a01, in whic al. 1984 proposedII complex I. DOI: 10.1107/S2414314621003977/vm4048Isup2.hklStructure factors: contains datablock(s) I. DOI: 2067247CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Three new styryl\u00adquinoline\u2013chalcone hybrids are been syntheized using a three-step reaction sequence. In two of them, a combination of hydro\u00adgen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions generates three-dimensional assemblies, but in the third, only a single weak \u03c0\u2013\u03c0 stacking inter\u00adaction is present, linking the mol\u00adecules into chains. 1H/13C NMR spectroscopy, and by high-resolution mass spectrometry, and the three products have been characterized by single-crystal X-ray diffraction. The mol\u00adecular conformations of (E)-3-{4-[(E)-2-phenyl\u00adethen\u00adyl]quinolin-2-yl}-1-(naph\u00adtha\u00adlen-1-yl)prop-2-en-1-one, C30H21NO, (IVa), and (E)-3-{4-[(E)-2-(4-fluoro\u00adphen\u00adyl)ethen\u00adyl]quinolin-2-yl}-1-(naph\u00adtha\u00adlen-1-yl)prop-2-en-1-one, C30H20FNO, (IVb), are very similar. In each com\u00adpound, the mol\u00adecules are linked into a three-dimensional array by hydro\u00adgen bonds, of the C\u2014H\u22efO and C\u2014H\u22efN types in (IVa), and of the C\u2014H\u22efO and C\u2014H\u22ef\u03c0 types in (IVb), and by two independent \u03c0\u2013\u03c0 stacking inter\u00adactions. By contrast, the conformation of the chalcone unit in (E)-3-{4-[(E)-2-(2-chloro\u00adphen\u00adyl)ethen\u00adyl]quinolin-2-yl}-1-(naph\u00adtha\u00adlen-1-yl)prop-2-en-1-one, C30H20ClNO, (IVc), differs from those in (IVa) and (IVb). There are only weak hydro\u00adgen bonds in the structure of (IVc), but a single rather weak \u03c0\u2013\u03c0 stacking inter\u00adaction links the mol\u00adecules into chains. Comparisons are made with some related structures.Three new styryl\u00adquinoline\u2013chalcone hybrids have been synthesized using a three-step pathway starting with Friedl\u00e4nder cyclo\u00adcondensation between (2-amino\u00adphen\u00adyl)chalcones and acetone to give 2-methyl-4-styryl\u00adquinolines, followed by selective oxidation to the 2-formyl analogues, and finally Claisen\u2013Schmidt condensation between the formyl inter\u00admediates and 1-acetyl\u00adnaph\u00adtha\u00adlene. All inter\u00admediates and the final products have been fully characterized by IR and Compound (IIc): reaction time 14\u2005h, yield 0.21\u2005g (73%), yellow solid, m.p. 388\u2013390\u2005K, RF\u00a0= 0.22 (12.5% ethyl acetate\u2013hexa\u00adne).Compound (IIa): reaction time 15\u2005h, yield 0.19\u2005g (86%), yellow solid, m.p. 367\u2013369\u2005K, v/v) to give the required formyl inter\u00admediates (IIIa)\u2013(IIIc) as solid com\u00adpounds.For the synthesis of com\u00adpounds (III), a suspension of the appropriate 2-methyl-4-styryl\u00adquinoline (II) (1.0\u2005mmol) and selenium dioxide (2.0\u2005mmol) in 1,4-dioxane (5\u2005ml) was stirred and heated at 373\u2005K for the appropriate time. After the complete consumption of (II) [as monitored by thin-layer chromatography (TLC)], di\u00adchloro\u00admethane (15\u2005ml) was added and the residual solid was removed by filtration. The solvent was removed under reduced pressure and the resulting crude products were purified by flash column chromatography on silica gel using hexa\u00adne\u2013ethyl acetate mixtures as eluent (compositions ranged from 7:1 to 2:1 RF\u00a0= 0.31 (9.1% ethyl acetate\u2013hexa\u00adne). Compound (IIIb): reaction time, 1\u2005h, yield 0.14\u2005g (89%), yellow solid, m.p. 417\u2013419\u2005K, RF\u00a0= 0.20 (9.1% ethyl acetate\u2013hexa\u00adne). Compound (IIIc): reaction time, 2\u2005h, yield 0.21\u2005g (92%), pale orange solid, m.p. 431\u2013433\u2005K, RF\u00a0= 0.28 (9.1% ethyl acetate\u2013hexa\u00adne).Compound (IIIa): reaction time, 1\u2005h, yield 0.23\u2005g (96%), yellow solid, m.p. 421\u2013423\u2005K, For the synthesis of com\u00adpounds (IV), a mixture of the appropriate 2-formyl inter\u00admediate (III) (1.0\u2005mmol), 1-aceto\u00adnaphthone (1.0\u2005mmol) and potassium hydroxide (1.1\u2005mmol) in ethanol (3\u2005ml) was stirred at 298\u2005K for the appropriate time. After complete consumption of (III) (monitored by TLC), the resulting precipitate was collected by filtration, washed with water (15\u2005ml) and ethanol (10\u2005ml), and then recrystallized from chloro\u00adform\u2013ethanol to afford the target mol\u00adecular hybrids (IV).RF\u00a0= 0.22 (13% ethyl acetate\u2013hexa\u00adne). Compound (IVb)RF\u00a0= 0.31 (13% ethyl acetate\u2013hexa\u00adne). Compound (IVc)RF\u00a0= 0.20 (9% ethyl acetate\u2013hexa\u00adne).Compound (IVa)supporting information.Full details of the spectroscopic characterization are included in the Uiso(H)\u00a0= 1.2Ueq(C).Crystal data, data collection and refinement details for com\u00adpounds (IVa)\u2013(IVc) are summarized in Table\u00a01et al., 20221H/13C NMR spectroscopy, and by high-resolution mass spectrometry (HRMS); full details of the spectroscopic characterization are provided in the supporting information.We have recently reported and (IIc) matched perfectly those of previously reported analogues \u2013(IVc). The formation of mol\u00adecular hybrids (IV) was established by disappearance of the formyl signals from both the 1H and 13C NMR spectra, and by the appearance of signals from the newly formed 3-aryl\u00adpropen-1-one fragment. As far as the Claisen\u2013Schmidt condensation is con\u00adcerned, it proceeded in a highly stereoselective manner, giving exclusively the E-stereoisomers, as indicated by the 1H NMR spectra. The trans configuration of the aryl\u00adpropen-1-one fragment was deduced on the basis of the coupling constant values between HA and HB , whose signals in the 1H NMR spectra appear at \u03b4 7.91\u20137.93 and 7.78\u20137.79, respectively.The presence of stretching vibration bands in the range 1727\u20131731\u2005cmE-configuration of both the styryl and the chalcone moieties \u2013(IVc) which fully confirm the mol\u00adecular structures deduced from the spectroscopic data, in particular, the s Figs. 1\u20133 \u25b8 \u25b8. et al., 2022et al., 2022ca 180\u00b0 around the C22\u2014C23 bond (Table\u00a02For each of (IVa)\u2013(IVc), the atom labelling Figs. 1\u20133 \u25b8 \u25b8 fet al., 1998abet al., 2000et al., 1990et al., 1995The supramolecular assembly in com\u00adpound (IVa)n, n) alternate with n, n), where n represents an integer in each case. The pyri\u00addine rings of the mol\u00adecules at and are strictly parallel with an inter\u00adplanar spacing of 3.2877\u2005(5)\u2005\u00c5 and a ring-centroid separation of 3.5372\u2005(7)\u2005\u00c5, corresponding to a ring-centroid offset of 1.305\u2005(2)\u2005\u00c5. This inter\u00adaction links the x, y, z) and the styryl ring at make an inter\u00adplanar angle of only 6.37\u2005(7)\u00b0; the ring-centroid separation is 3.7818\u2005(9)\u2005\u00c5 and the shortest perpendicular distance between the centroid of one ring and the plane of the other is 3.4535\u2005(6)\u2005\u00c5, corresponding to a ring-centroid offset of 1.541\u2005(2)\u2005\u00c5. This inter\u00adaction links the The linking of these dimeric units by C\u2014H\u22efN hydro\u00adgen bonds gives rise to a ribbon running parallel to the and quinoline, the mol\u00adecules are linked into cyclic centrosymmetric dimers by hydro\u00adgen bonds, of the C\u2014H\u22efN and C\u2014H\u22ef\u03c0 types, respectively, and these dimers are further linked by \u03c0\u2013\u03c0 stacking inter\u00adactions to form sheets in the fluoro com\u00adpound and chains in the tri\u00adfluoro\u00admethyl analogue. By contrast, there are no significant inter\u00admolecular inter\u00adactions in the structure of (E)-4--2-methyl\u00adquinoline. All of these type (II) com\u00adpounds have mol\u00adecular skeletons in which the styryl and quinoline units are non-coplanar, as reported here for com\u00adpounds (IVa)\u2013(IVc). This appears to be the case for all of the 4-styryl\u00adquinolines which have been structurally characterized so far, in contrast to the 2- and 8-styryl\u00adquinolines, where the two ring systems appear always to be effectively coplanar three products and all of the inter\u00admediates on the pathways leading to them, and we have determined the mol\u00adecular and supra\u00admolecular structures of the three products.We have developed a highly versatile and efficient three-step synthesis of a novel class of styryl\u00adquinoline\u2013chalcone hybrids based on only very simple and readily available starting materials, such as simple aldehydes and ketones, and we have characterized by spectroscopic means global, IVa, IVb, IVc. DOI: 10.1107/S2053229622011263/ov3164IVasup2.hklStructure factors: contains datablock(s) IVa. DOI: 10.1107/S2053229622011263/ov3164IVbsup3.hklStructure factors: contains datablock(s) IVb. DOI: 10.1107/S2053229622011263/ov3164IVcsup4.hklStructure factors: contains datablock(s) IVc. DOI: 10.1107/S2053229622011263/ov3164sup5.txtSpectroscopic data. DOI: 2221749, 2221750, 2221751CCDC references:"} +{"text": "A and B) are present in the asymmetric unit, with different conformations. The dihedral angle between the mean planes of the carbazole systems for mol\u00adecule A is 49.1\u2005(2)\u00b0 compared to 84.0\u2005(1)\u00b0 for mol\u00adecule B. In the crystal, numerous aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions [shortest centroid\u2013centroid separation = 3.7069\u2005(19)\u2005\u00c5] help to establish the three-dimensional supra\u00admolecular network.In the title compound, two independent mol\u00adecules are present in the asymmetric unit, with different conformations. The dihedral angle between the mean planes of the carbazole systems for mol\u00adecule A is 49.1\u2005(2)\u00b0 compared to 84.0\u2005(1)\u00b0 for mol\u00adecule B. In the crystal, numerous aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions [shortest centroid\u2013centroid separation = 3.7069\u2005(19)\u2005\u00c5] help to establish the three-dimensional supra\u00admolecular network.In the title compound, C Two independent mol\u00adecules (A and B) are present in the asymmetric unit, with different conformations. Each of the independent mol\u00adecules is composed of two carbazole systems connected by an iodo\u00adbenzene bridge \u00b0 compared to 84.0\u2005(1)\u00b0 for the N3 and N4 ring systems in mol\u00adecule B. In the crystal, numerous face-to-face [shortest centroid\u2013centroid separation = 3.7069\u2005(19)\u2005\u00c5] and edge-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions connect the mol\u00adecules into a three-dimensional supra\u00admolecular network , t-BuOK , and dimethyl sulfoxide (10\u2005ml) was stirred at 120\u00b0C before 1-iodo-3,5-di\u00adfluoro\u00adbenzene was injected. The reaction mixture was stirred at 140\u00b0C for 2\u2005h. After being cooled, the mixture was extracted with chloro\u00adform and the organic extracts were combined, washed with water, and the organic layer was dried over anhydrous MgSO4. After evaporating the solvent, the crude product was purified by column chromatography on silica gel with chloro\u00adform/n-hexane as the eluent to give a white powder. Yield: 83%. 1H NMR \u03b4 8.17 , 8.08 , 7.84 , 7.56 , 7.49 , 7.35 . Colourless blocks were obtained by recrystallization from mixed solvents of methyl\u00adene chloride and n-hexane (v:v = 1.2).A mixture of 9Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2414314621004284/hb4376sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314621004284/hb4376Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314621004284/hb4376Isup3.cmlSupporting information file. DOI: 2079137CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Nickel(II) Schiff base complexes containing thiol\u00adate S and polyamine N donor atoms exhibit electrocatalytic activity for proton reduction. The piperazine moiety in the Schiff base ligand gives a smaller bite angle, which is effective in reducing the overpotential. N)(meth\u00adyl)amino-\u03baN]eth\u00adyl}imino-\u03baN)meth\u00adyl]benzene\u00adthiol\u00adato-\u03baS}nickel(II) chloride, [Ni(C12H18N3S)]Cl (1), and [2-eth\u00adyl]imino-\u03baN}meth\u00adyl)benzene\u00adthiol\u00adato-\u03baS]nickel(II) hexa\u00adfluoro\u00adphosphate di\u00adchloro\u00admethane monosolvate, [Ni(C13H18N3S)]PF6\u00b7CH2Cl2 (2), were synthesized by the reactions of 2-(tert-butyl\u00adthio)\u00adbenzaldehyde, tri\u00adamines, and nickel(II) salts. Both complexes have a nickel ion surrounded by an N,N\u2032,N\u2032\u2032,S-tetra\u00addentate ligand, forming a square-planar geometry. The terminal N,N-chelating moiety is N,N-di\u00adalkyl\u00adethane-1,2-di\u00adamine for 1 and 1-alkyl\u00adpiperazine for 2. The N\u2014Ni\u2014N bite angle in the terminal N,N-chelate ring in 2 [76.05\u2005(10)\u00b0] is much smaller than that in 1 [86.16\u2005(6)\u00b0]. Cyclic voltammograms of 1 and 2 in aqueous media indicated that the reduction and oxidation potentials of 2 are more positive than those of 1. The smaller bite angle of the terminal piperazine chelate in 2 reduces the electron-donating ability of the tetra\u00addentate ligand, resulting in a positive shift of the redox potentials. Both complexes exhibit catalytic activity for proton reduction, and the piperazine moiety in 2 is effective in reducing the overpotential.The thiol\u00adate nickel complexes {2-[({2-[(2-amino\u00adethyl-\u03ba In the electrocatalytic proton reduction, the electrochemical properties of the precatalysts are directly related to the formation of real catalysts. Therefore the tuning of the redox properties is also required in the ligand design.The nickel(II) complex Cl (1) and [Ni(C13H18N3S)]PF6 (2), in which the N,N\u2032,N\u2032\u2032,S-tetra\u00addentate Schiff base ligands contain an additional N-methyl group and a terminal piperazine moiety, respectively. The electrochemical properties of these complexes were investigated by cyclic voltammetry in water, and compared with those of [Ni(C11H16N3S)]Cl (3) without N-substituents.In this work we synthesized two water-soluble N2.1 and 2 consist of an Ni2+ ion and a monoanionic N,N\u2032,N\u2032\u2032,S-tetra\u00addentate ligand, giving a square-planar geometry. The asymmetric unit in 1 comprises the complex cation and a chloride anion, whereas in 2 a hexa\u00adfluoro\u00adphosphate anion and a di\u00adchloro\u00admethane mol\u00adecule are incorporated into the crystal lattice.The complex cations in N,S-chelate, the N1\u2014Ni1\u2014S1 angles are 97.77\u2005(5)\u00b0 in 1 (Table\u00a012 (Table\u00a023 [98.5\u2005(2)\u00b0] and the tetra\u00adphenyl\u00adborate salt [Ni(C11H16N3S)]B(C6H5)4 . The bond distances in the chelate rings are also comparable \u00b0 in 1, 87.80\u2005(10)\u00b0 in 2, 86.1\u2005(3)\u00b0 in 3 complexes with S,N,N,S-tetra\u00addentate ligands, which have two amine N or two imine N donor atoms besides two S atoms \u00b0] is much smaller than those of 1 [86.16\u2005(6)\u00b0], 3 and 2 [N1\u2014C8\u2014C9\u2014N2 = 45.1\u2005(3)\u00b0], this bending is due to the rigid structure of the piperazine chelate, which fixes the direction of the methyl\u00adene groups on the tertiary amine N atom.In complex 3.1 shows hydrogen bonds between the terminal amine nitro\u00adgen atom in the complex cation and two chloride ions with the N3\u22efCl1 and N3\u22efCl1 and 3.8965\u2005(13)\u2005\u00c5].The crystal structure of y Table\u00a03, which aon Fig.\u00a03. The dis2 between the methyl\u00adene hydrogen atoms of the polyamine moiety and the \u03c0 system of the benzene ring distance of 3.114\u2005(3)\u2005\u00c5 = 2.47\u2005(4)\u2005\u00c5].Several inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions exist in g Table\u00a04. The pip\u00c5 Table\u00a04. In addi4.N,N\u2032,N\u2032\u2032,S-tetra\u00addentate Schiff base ligands studied here have not been reported so far for other transition-metal ions. A similar Schiff base structure that contains benzene\u00adthiol\u00adate and polyamines is found in a trinuclear nickel(II) complex with a C3-symmetric ligand based on a 1,3,5-trimercapto\u00adbenzene backbone \u00b0 complexes 1, 2, and 3 was investigated by cyclic voltammetry. Measurements were performed in 5 \u00d7 10\u22124M (1\u2005M = 1\u2005mol\u2005dm\u22123) aqueous solution containing KNO3 (0.1\u2005M) at a scan rate of 0.1\u2005V\u2005s\u22121. The working electrode was a glassy carbon disk electrode with a diameter of 3\u2005mm, the auxiliary electrode was a platinum wire, and the reference electrode was Ag/AgCl/saturated KCl. All complexes exhibit irreversible reduction and oxidation processes suggest the adsorption of the reduced species. In the oxidation process, the anodic wave appeared at 0.73\u2005V for 1, 0.79\u2005V for 2, and 0.68\u2005V for 3. In both processes, the redox potentials of 1 are slightly shifted to more positive values than those of 3, which suggests that the electronic and steric effects of the methyl group on the central N atom are not so significant. The voltammogram of 2 shows further positive shifts, and the shift in the reduction process is more pronounced. This is probably related to the smaller bite angle of the terminal piperazine chelate, which reduces the electron-donating ability of the Schiff base ligand toward the nickel center.The redox behavior of the es Fig.\u00a05. In the 1 and 2 were compared in a buffer solution of pH 4.6 (0.1 M acetic acid/sodium acetate). A catalytic current was observed during the reduction process, giving a peak at \u22121.28\u2005V for 1 and \u22121.23\u2005V for 2 under N2. Elemental analyses were performed by the Analytical Research Service Center at Osaka City University or A Rabbit Science Co., Ltd.12H18N3S)]Cl (1).PF6 (2).[Ni(C To a solution of N-(2-amino\u00adeth\u00adyl)piperazine and 2-(t-butyl\u00adthio)\u00adbenzaldehyde in ethanol (20\u2005mL) was added NiCl2\u00b76H2O . The resulting suspension was refluxed under a nitro\u00adgen atmosphere for 8\u2005h, during which time the color of the solution turned orange and a yellow\u2013green precipitate formed. The reaction mixture was filtered, and an ethanol solution (5\u2005mL) of ammonium hexa\u00adfluoro\u00adphosphate was added to the filtrate. The resulting orange precipitate was collected by filtration and dried under reduced pressure to give an orange powder of 2 . Suitable crystals for X-ray diffraction analysis were grown from a di\u00adchloro\u00admethane solution by layering with diethyl ether. 1H NMR : \u03b4 2.76\u20132.94 2NH), 2.83 , 3.91\u20134.03 2NH), 4.16\u20134.27 2NH), 4.31 , 7.10 , 7.28 , 7.52 , 7.74 , 8.16 . Analysis calculated for C13H18F6N3NiPS\u00b70.75CH2Cl2: C, 32.02; H, 3.81; N, 8.15. Found: C, 32.11; H, 3.93; N, 8.14.8.N,N-chelating moieties in 1 were modeled as disordered over two positions each, and the occupancy factors refined to 0.864\u2005(3) and 0.136\u2005(3). Hydrogen atoms on the disordered C atoms and the adjacent C atoms that belong to the minor site were placed in calculated positions with C\u2014H(meth\u00adyl) = 0.98\u2005\u00c5 and C\u2014H(methyl\u00adene) = 0.99\u2005\u00c5 and refined using a riding model with Uiso(H) = 1.5Ueq(C) and 1.2Ueq(C), respectively. Other H atoms were found in a difference-Fourier map and freely refined.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989022003954/dj2045sup1.cifCrystal structure: contains datablock(s) 1, 2. DOI: 10.1107/S2056989022003954/dj20451sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989022003954/dj20452sup3.hklStructure factors: contains datablock(s) 2. DOI: 2165891, 2165890CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II ions both show a distorted N3Cl3 octa\u00adhedral coordination environment defined by three N atoms of the tridentate 2,4,6-tri-2-pyridyl-1,3,5-triazine ligand, two bridging Cl\u2212 ligands and a Cl\u2212 anion.The Ni 2Cl4(C18H12N6)2], the NiII ions are hexa-coordinated in a distorted octa\u00adhedral coordination environment defined by three N atoms of the tridentate 2,4,6-tri-2-pyridyl-1,3,5-triazine ligand and three Cl\u2212 anions in a meridional geometry. The two NiII ions are bridged by two Cl anionic ligands, thereby forming a dinuclear complex. A crystallographic centre of inversion is located at the centroid of the Ni2Cl2 ring.In the title compound, [Ni On the other hand the Cl2\u2014Ni1\u2014Cl1i axis (symmetry code: (i) \u2212x, \u2212y\u00a0+\u00a01, \u2212z) is almost linear [Cl2\u2014Ni1\u2014Cl1i = 176.25\u2005(2)\u00b0]. The Ni\u2014N(pyrid\u00adyl) bonds [Ni1\u2014N4/N6 = 2.130\u2005(2) and 2.129\u2005(2)\u2005\u00c5] are slightly longer than the Ni\u2014N(triazine) bond [Ni1\u2014N1 = 1.970\u2005(2)\u2005\u00c5]. The three Ni\u2014Cl bond lengths are somewhat different . The two pyridyl rings that coordinat to the NiII atom are located approximately parallel to the respective triazine ring, making dihedral angles of 4.51\u2005(6) and 4.95\u2005(6)\u00b0, respectively. The dihedral angle between the non-coordinating pyridyl substituent and the triazine ring is 7.56\u2005(6)\u00b0.In the complex, the two Niex Fig.\u00a01. Each NiCg1 (the centroid of ring N5/C8\u2013C12) and Cg2ii [the centroid of ring N6/C14\u2013C18; symmetry code: (ii) x, \u2212y\u00a0+\u00a0z\u00a0\u2212\u00a0The complex displays numerous inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between adjacent six-membered rings. For e Table\u00a01.2\u00b76 H2O in ethanol (30\u2005ml) was added 2,4,6-tri-2-pyridyl-1,3,5-triazine . The solution was stirred for 12\u2005h at room temperature. The formed precipitate was separated by filtration, washed with ethanol and acetone, and dried at 323\u2005K, to give a pale-green powder . Brown crystals suitable for X-ray analysis were obtained by slow evaporation from a dimethyl sulfoxide (DMSO) solution at 363\u2005K.To a solution of NiCl\u22123) and minimum (\u22120.20\u2005e\u2005\u00c5\u22123) electron density in the difference Fourier map are located 0.73 and 1.28\u2005\u00c5, respectively, from atoms C2 and C9.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621000936/im4011sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314621000936/im4011Isup2.hklStructure factors: contains datablock(s) I. DOI: 2058987CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The units are further linked by O\u2014H\u22efO and N\u2014H\u22efCl hydrogen bonds.In the crystal of the title organic salt, the cation and the anion are connected H-benzimidazol-3-ium] 2,5-di\u00adchloro-3,6-dioxo\u00adcyclo\u00adhexa-1,4-diene-1,4-diolate}, 2C13H11N2O+\u00b7C6Cl2O42\u2212, the chloranilate anion is located on an inversion centre, so that the asymmetric unit contains one cation and one half of the chloranilate anion. In the crystal, the cation and the anion are connected by a bifurcated N\u2014H\u22ef hydrogen bond, forming a 2:1 unit. The units are linked into a layer lying parallel to -1 D\u2014H\u22efA hydrogen bonding in chloranilic acid\u2013organic base systems ; symmetry code as given in Table\u00a01via O\u2014H\u22efO and N\u2014H\u22efCl hydrogen bonds \u2005\u00c5 and C2\u2014Cl1\u22efCg3iv = 139.21\u2005(5)\u00b0; symmetry code: (iv) x, y, z\u00a0\u2212\u00a01] is observed between the layers, where Cg3 is the centroid of the C11\u2013C16 ring.We have prepared the title compound in order to continue our studies of H-benzimidazole in a ca 1:1 molar ratio at room temperature [150\u2005ml methanol solution of chloranilic acid (0.45\u2005g) and 2-(2-hy\u00addroxy\u00adphen\u00adyl)-1H-benzimidazole (0.45\u2005g)].Single crystals of the title salt were obtained by slow evaporation from a methanol solution of chloranilic acid with 2-(2-hy\u00addroxy\u00adphen\u00adyl)-1Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621011500/hb4396sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314621011500/hb4396Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314621011500/hb4396Isup3.cmlSupporting information file. DOI: 2119367CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Targeted drug delivery systems hold the remarkable potential to improve the therapeutic index of anticancer medications markedly. Here, we report a targeted delivery platform for cancer treatment using clathrin light chain (CLC)\u2010conjugated drugs. We conjugated CLC to paclitaxel (PTX) through a glutaric anhydride at high efficiency. Labeled CLCs localized to 4T1 tumors implanted in mice, and conjugation of PTX to CLC enhanced its delivery to these tumors. Treatment of three different mouse models of cancer\u2014melanoma, breast cancer, and lung cancer\u2014with CLC\u2010PTX resulted in significant growth inhibition of both the primary tumor and metastatic lesions, as compared to treatment with free PTX. CLC\u2010PTX treatment caused a marked increase in apoptosis of tumor cells and reduction of tumor angiogenesis. Our data suggested HSP70 as a binding partner for CLC. Our study demonstrates that CLC\u2010based drug\u2010conjugates constitute a novel drug delivery platform that can augment the effects of chemotherapeutics in treating a variety of cancers. Moreover, conjugation of therapeutics with CLC may be used as means by which drugs are delivered specifically to primary tumors and metastatic lesions, thereby prolonging the survival of cancer patients. ACNacetonitrileADCantibody\u2010drug conjugateBUNblood urea nitrogenCD31cluster of differentiation 31CLCclathrin light chainDAPIdiamidino\u20102\u2010phenylindoleDLSdynamic light scatteringDMEMDulbecco's modified Eagle's mediumDMSOdimethyl sulfoxideECMextracellular matrixEDC1\u2010ethyl\u20103\u2010(3\u2010dimethylaminopropyl)carbodiimideESIMSelectron spay ionization mass spectrometryE. coliEscherichia coliFBSfetal bovine serumGAPDHglyceraldehyde\u20103\u2010phosphate dehydrogenaseHPLChigh\u2010performance liquid chromatographyHRPhorseradish peroxidaseHSP70heat shock protein 70H&Ehematoxylin and eosinLLC1Lewis lung carcinoma 1LYVE1lymphatic vessel endothelial hyaluronan receptor 1MALDI\u2010TOFmatrix\u2010assisted laser desorption ionization time\u2010of\u2010flightMFImean fluorescence intensityMSmass spectrometryNHSN\u2010hydroxysuccinimideNMRnuclear magnetic resonancePDBprotein data bankPKpharmacokineticsPTXpaclitaxelROIregion of interestRPMIRoswell park memorial instituteSDstandard deviationSDS\u2010PAGEsodium dodecyl sulfate\u2010polyacrylamide gel electrophoresissiRNAsmall interfering ribonucleic acidTDLNtumor\u2010draining lymph nodeTGItumor growth inhibitionUV\u2013Visultraviolet\u2013visible1Targeted drug delivery systems can amplify the concentration of transported payloads at various tissues of interest, including tumors.Antibody\u2010drug conjugates (ADCs) are emerging platforms for the delivery of a range of cancer drugs.Here, we report a novel application of clathrin light chain A (CLC)\u2010based drug delivery. CLC is an endogenous small protein that forms a network of triskelions that constitute a polyhedral lattice around vesicles that assist in the sorting of cargo for intracellular trafficking.First, we confirmed that CLCs interact with cancer cells and could be used as vehicles for targeted delivery to malignant tumors. Next, CLCs were conjugated to the antineoplastic agent paclitaxel (PTX) via reaction with glutaric anhydride (CLC\u2010PTX) to evaluate their utility for cancer\u2010targeted delivery and treatment in a series of mouse models. CLC\u2010PTX enhanced the concentration of PTX at the tumor site and suppressed tumor growth in mouse models of breast cancer, melanoma, and lung carcinoma. In addition, CLC\u2010PTX was found to reduce the size of metastatic lesions in breast cancer.22.1Escherichia coli (E. coli), confirmed by sodium dodecyl sulfate\u2010polyacrylamide gel electrophoresis (SDS\u2010PAGE), and extracted at a purity of >85%, as demonstrated by Western blot (data not shown). PTX is a chemotherapeutic drug used for the treatment of numerous cancers.1H\u2010NMR was expressed in R Figure\u00a0 and 13C\u2010R Figure\u00a0. In the N\u2010hydroxysuccinimide (NHS) coupling to produce CLC\u2010PTX*, which was verified by ultraviolet\u2013visible (UV\u2013Vis) spectroscopy was used to confirm the conjugation of PTX to CLC. The maximal absorbance of PTX* is observed at a wavelength of around 500\u2009nm. We conjugated 2\u2032\u2010glutaryl PTX* to CLC via 1\u2010ethyl\u20103\u2010(3\u2010dimethylaminopropyl)carbodiimide (EDC)/sulfo\u2010y Figure\u00a0. The absy Figure\u00a0. After cy Figure\u00a0, whereasy Figure\u00a0. Next, Cy Figure\u00a0. Intravey Figure\u00a0A. In addy Figure\u00a0B and C.2.2We sought to determine whether CLC was internalized by 4T1 mouse breast cancer cells by incubating these cells with CLCs attached to Alexa Fluor\u2122 488 dye. The colocalization of the CLCs with a lysosome marker was confirmed Figure\u00a0, which iA virtual screening between CLC and a protein data bank (PDB) was conducted, resulting in the identification of three proteins with high binding energy Table\u00a0. Interes2.3iv to 4T1 tumor\u2010bearing BALB/c mice. The mice were euthanized at 1\u00a0day (1\u00a0d), 2\u2009days, and 3\u2009days following administration of CLCs, at which time points we measured the fluorescent signal of CLC\u2010IR800 in various organs, using an iBox Exploreriv with CLC\u2010PTX* and quantified the fluorescence of PTX*. PTX* did not clear significantly from the tumor within the span of the study, whereas the PTX* signal declined in the liver and the kidney over time e Figure\u00a0.The lung is the most common site of metastasis for breast cancer. Lung with 4T1 metastasis showed enhanced targeting signals of CLC as compared with na\u00efve lung Figure\u00a0. Interesiv administration to 4T1 breast tumor\u2010bearing BALB/c mice. Ex vivo fluorescent images of the tumors were acquired 1\u00a0day following administration of PTX* or CLC\u2010PTX* at an equivalent PTX* dose of 0.5\u00a0mg/kg, which showed that the accumulation of CLC\u2010PTX* in the tumor was higher than free PTX* quantified PTX. We detected significant amounts of PTX in the sera of the CLC\u2010PTX group as compared with the free PTX group at 6\u00a0h, indicating that conjugation to CLC enhanced the blood circulation of PTX . The tumors in the CLC\u2010PTX group displayed slower tumor progression in comparison to the two other treatment groups of 4T1 tumor\u2010bearing mice at 27\u2009days following implantation Figure\u00a0. The exp2.63, following a schedule and dosing identical to the 4T1 study above. Again, CLC\u2010PTX displayed the most potent inhibitory effect on tumor growth: at the end of the study, the tumors of the mice treated iv with CLC\u2010PTX were the smallest (0.81\u2009\u00b1\u20090.30\u2009\u00d7\u2009103\u2009mm3), as compared to the PBS\u2010treated mice (2.36\u2009\u00b1\u20090.61\u2009\u00d7\u2009103\u2009mm3) and free PTX\u2010treated mice (1.29\u2009\u00b1\u20090.50\u2009\u00d7\u2009103\u2009mm3) . Immunostaining demonstrated a decline of Melan\u2010A+ cells with an increase in caspase\u20103 expression in the CLC\u2010PTX\u2010treated mice as compared to other groups Figure\u00a0. TGI by s Figure\u00a0. The B16s Figure\u00a0.2.7iv to C57BL/6 mice bearing LLC1 murine lung carcinoma, following a schedule identical and dosing in the aforementioned tumor models. The tumors in the CLC\u2010PTX group (0.80\u2009\u00b1\u20090.21\u2009\u00d7\u2009103\u2009mm3) were significantly smaller than those in the mice that received PBS (1.68\u2009\u00b1\u20090.30\u2009\u00d7\u2009103\u2009mm3) and free PTX (1.21\u2009\u00b1\u20090.31\u2009\u00d7\u2009103\u2009mm3) . Next, we stained LLC1 tumor tissue sections with the cancer marker pan\u2010cytokeratin, apoptosis marker caspase\u20103, proliferation marker Ki67, and vascular marker CD31, and examined them by fluorescence microscopy. Treatment with CLC\u2010PTX reduced the amount of Pan\u2010Cytokeratin+ cancer cells significantly, whereas Caspase\u20103+ apoptotic cells increased Figure\u00a0. In addid Figure\u00a0. Consistd Figure\u00a0. Furtherd Figure\u00a0. Thus, t3Targeted drug delivery systems have attracted major interest and already entered into clinical practice for various diseases, but their application to cancer remains to be developed.Metastasis is the primary factor of cancer morbidity and mortality.Nonetheless, future in vivo targeting studies are required to define the mechanisms by which HSP70 functions as a target of CLC\u2010conjugated drug delivery. In addition, HSP70 produced by tumor cells may undergo differential glycosylation, so kinetic studies tailored to each type of cancer may be required for more specific and accurate assessment of the binding affinity of HSP70 to CLC. The performance of a mutational study to identify putative binding site of CLCOur data suggest that CLC targeted the primary tumor and metastatic lung lesions of 4T1 breast cancer in mice with high efficacy. While there was an increase in CLCs in the peripheral organs, in particular, the kidney; however, this retention faded after 3\u2009days. Nonetheless, the tumor uptake of exogenously administered CLCs remains relatively stable over time.CLC\u2010PTX enhanced the therapeutic efficacy of PTX on the primary 4T1 tumor, as demonstrated by suppression of tumor growth, enhancement of cancer cell apoptosis, and especially inhibition of angiogenesis. CLC\u2010PTX treatment was also found to reduce the density of lymphatic vasculature, which often contributes to the immunosuppressive environment of TDLN. Concomitantly, distant lung metastasis was also inhibited more effectively by treatment with CLC\u2010PTX than free PTX. Excessive accumulation of extracellular matrix (ECM) in the tumor stroma is referred to as the desmoplastic reaction, commonly reported as a major obstacle to the treatment of cancers.44.14T1 mouse breast cancer (ATCC\u00ae CRL\u20102539\u2122), B16 mouse melanoma (ATCC\u00ae CRL\u20106322\u2122), LLC1 mouse lung carcinoma (ATCC\u00ae CRL\u20101642\u2122), and HK02 human kidney tubular epithelial cell lines (ATCC\u00ae CRL\u20102190\u2122) were purchased from American Type Culture Collection . 4T1, B16, LLC1, and HK02 cells were cultured in RPMI\u20101640 or Dulbeccos modified Eagles medium with 10% fetal bovine serum and 1% penicillin/streptomycin (pen/strep).4.2C57BL/6 (JAX# 000664) and BALB/c (JAX# 000651) mice were obtained from The Jackson Laboratories. All animal experiments and methods were performed in accordance with the relevant guidelines and regulations approved by the Institutional Animal Care and Use Committee (protocols: 2016\u2009N000167/04977) of Brigham and Women's Hospital .4.3E. coli expression system as follows. The human CLC gene was codon\u2010optimized for expression in E. coli and chemically synthesized by Biomatik. The synthesized gene was subsequently cloned into NdeI/HindIII sites with pET30a vector. These expression plasmids were used to transform the E. coli BL21(DE3). His\u2010tag was applied to a Ni\u2010NTA column and eluted in a buffer containing 10\u2009mM Tris, 0.15\u2009M NaCl, 8\u00a0M urea, and 0.3\u00a0M imidazole at a pH of 8.0.Human CLC was expressed and optimized for an Gene sequence; ATGGCGGAACTGGACCCGTTCGGCGCTCCGGCAGGCGCACCGGGCGGTCCGGCGCTGGGTAACGGCGTTGCGGGTGCTGGTGAAGAAGACCCGGCAGCAGCGTTCCTGGCGCAGCAGGAATCTGAAATCGCAGGTATCGAAAACGATGAAGCGTTCGCGATCCTGGACGGTGGTGCTCCGGGTCCGCAGCCGCACGGTGAACCGCCGGGTGGTCCGGATGCGGTTGACGGTGTTATGAACGGCGAGTACTACCAGGAGTCTAACGGTCCGACCGATTCTTACGCGGCAATTAGCCAGGTTGATCGTCTGCAATCCGAACCGGAATCTATCCGTAAATGGCGTGAGGAGCAGATGGAACGCCTGGAAGCTCTGGACGCGAACTCTCGCAAACAGGAGGCGGAATGGAAAGAAAAAGCGATCAAAGAGCTGGAAGAATGGTATGCGCGTCAGGACGAACAGCTGCAAAAAACCAAAGCGAACAACCGTGTGGCGGACGAAGCATTCTACAAACAGCCGTTTGCGGACGTTATCGGTTACGTTACCAACATCAACCATCCGTGCTACTCTCTGGAGCAGGCAGCGGAAGAAGCGTTCGTGAACGACATCGACGAATCTAGCCCAGGCACCGAATGGGAACGTGTTGCGCGCCTGTGCGACTTCAACCCGAAATCTTCTAAACAGGCTAAAGACGTTTCTCGTATGCGTTCTGTTCTGATCTCTCTGAAGCAGGCTCCGCTGGTTCAC.Amino sequence; MAELDPFGAPAGAPGGPALGNGVAGAGEEDPAAAFLAQQESEIAGIENDEAFAILDGGAPGPQPHGEPPGGPDAVDGVMNGEYYQESNGPTDSYAAISQVDRLQSEPESIRKWREEQMERLEALDANSRKQEAEWKEKAIKELEEWYARQDEQLQKTKANNRVADEAFYKQPFADVIYVTNINHPCYSLEQAAEEAFVNDIDESSPGTEWERVARLCDFNPKSSKQAKDVSRMRSVLISLKQAPLVH.The purity of CLC was >85%, as estimated by a Coomassie blue\u2010stained SDS\u2010PAGE gel. The concentration of CLC was determined by Bradford protein assay, using BSA as a standard. Its molecular weight was 28.1\u00a0kDa, and its isoelectric point was 4.37.4.4www.rcsb.org. Three proteins with the highest binding energy were identified through AutoDock Vina: ADP\u2010ribosylation factor 3, HSP70, and serine/threonine\u2010protein phosphatase PP1\u2010beta catalytic subunit was labeled with Alexa Fluor\u2122 488 NHS Ester (Thermo Fisher Scientific) or Alexa Fluor\u2122 594 NHS Ester (Thermo Fisher Scientific) and incubated with 4T1 cells for 2\u00a0h at 37\u00b0C. After five washes with PBS buffer, 4T1 cells were stained with lysosomal staining kit . 4T1 cells were fixed with 4% paraformaldehyde (Electron Microscopy Sciences), and diamidino\u20102\u2010phenylindole (DAPI) was used to counterstain the cell nuclei. The cells were visualized using an EVOS\u2122 FL Auto 2 Imaging System (Thermo Fisher Scientific). After splitting each color channel, the area with fluorescence was measured for each color channel with Image J . The percent of uptake of CLC was calculated by dividing with measured area from DAPI channel.4.62O with 0.1% formic acid for 20\u2009min), using a Phenomenex Luna 5\u00a0\u03bcm C18 column . The product was detected at 14.1\u00a0min, and molecular weight was confirmed as 969\u2009g/mol by electron spay ionization mass spectrometry analysis ([M\u2010H]\u2212 m/z at 968]). 2'\u2010Glutaryl PTX was purified by reversed\u2010phase HPLC with a gradient solvent system (15%\u201375% ACN/H2O with 0.1% formic acid for 40\u2009min). The product was eluted at a retention time of 13.4\u00a0min under these HPLC conditions. 2\u2032\u2010Glutaryl PTX was confirmed by 1H\u2010NMR (proton nuclear magnetic resonance) and 13C\u2010NMR (carbon nuclear magnetic resonance) spectra. 2'\u2010glutaryl PTX (0.2\u00a0mg) dissolved in dimethyl sulfoxide (DMSO) (Thermo Scientific Fisher) was activated with 1\u2010ethyl\u20103\u2010(3\u2010dimethylaminopropyl)carbodiimide hydrochloride and Sulfo\u2010NHS (N\u2010hydroxysulfosuccinimide) for 15\u2009min at RT in 2\u2010(N\u2010morpholino)ethanesulfonic acid (MES) buffer . The EDC was quenched by 2\u2010mercaptoethanol for 10\u00a0min. Immediately, the pH of the solution was increased by NaHCO3 to ~8. CLC dissolved in PBS was mixed with the activated 2\u2032\u2010glutaryl PTX for 2\u00a0h at RT . Dialysis was performed twice by a centrifugal filter at 10,000\u2009rpm for 15\u2009min to remove the free PTX. The solution was purified further by a desalting column .Glutaric anhydride and PTX were prepared in a 4\u00a0ml vial dried under high vacuum for 24\u2009h and dissolved in 1\u00a0ml of pyridine. The solution was stirred at room temperature under Ar atmosphere for 2\u00a0h. The mixture was diluted with 300\u2009\u03bcl of methanol, and 5\u00a0\u03bcl of solution was injected into liquid chromatography/mass spectrometery (LC/MS) with a gradient reversed\u2010phase system (10%\u2013100% ACN/H4.7280\u2009nm) of CLC was extracted from the absorption spectrum of CLC solutions at different concentrations. The \u03b5CLC,280\u2009nm was 4.8\u2009\u00d7\u2009104, whereas \u03b5PTX,500\u2009nm was 4.2\u2009\u00d7\u2009104. The concentration of CLC (CCLC) in the CLC\u2010PTX* solution was calculated by /\u03b5CLC,280\u2009nm, and CPTX* was by .Absorbance was measured by UV\u2013Vis spectroscopy . The molar extinction coefficient at 1:1 ratio. The 2\u2009\u03bcl of the samples was spotted, air\u2010dried, and detected with a linear detector. The data were analyzed using Bruker Daltonics flexAnalysis. External calibration was performed with BSA with [M\u2010H] average mass (m/z) of 66,428.MALDI mass analyses of CLC and CLC\u2010PTX were performed with a Bruker MicroFlex\u2122 MALDI\u2010TOF in negative mode. The protein solution was mixed with sinapinic acid matrix solution was incubated in acetate buffer or phosphate buffer at 37\u00b0C. The solutions were dialyzed with 5% DMSO solution, and filtrates were analyzed by UV\u2013Vis spectroscopy to quantify the amount of free PTX* in comparison to the absorbance of the solution prior to incubation.4.105) was stained with CLC\u2010Alexa 594 or anti\u2010HSP70\u2010Alexa 594 in FACS buffer at 4\u00b0C for 30\u2009min. Cells were washed by FACS buffer and fixed with FACS buffer containing 1% formalin. The autofluorescence from the cell itself was unmixed and only single cells were gated (Cytek\u00ae Aurora). Analysis of flow cytometry data was performed by FlowJo software . The histogram over Alexa 594 channel was extracted and the total fluorescence signal was divided by the total number of the cell.The surface of 4T1 cells (1.0\u00a0\u00d7\u2009104.11Cells were transfected with Lipofectamine 2000 (Invitrogen), according to the manufacturer's instructions. Briefly, cells were plated at 20%\u201330% density in 12\u2010well plates 24\u2009h prior to transfection. For siRNA transfection, the equivalent of 200\u2009nM of siRNA per well of a 12\u2010well plate was utilized. After a 48\u2009h incubation period, the lysates of the cells were measured using the Bradford assay. Equal amounts of protein were separated by SDS\u2010PAGE and transferred to a polyvinylidene fluoride (PVDF) membrane. The membranes were immunoblotted with the following specific antibodies: anti\u2010HSP70, anti\u2010rabbit IgG\u2010HRP (Sigma\u2010Aldrich), anti\u2010rat IgG\u2010HRP (Sigma\u2010Aldrich), and anti\u2010GAPDH (Sigma\u2010Aldrich), using standard protocols. The blots were developed with West Dura chemiluminescent substrates using a Bio\u2010Rad ChemiDoc imaging system.4.12s.c. with 1.0\u2009\u00d7\u2009105 4T1 cells. At 14\u2009days postimplantation, these mice received a single iv injection of CLC\u2010IR800 conjugate (4\u00a0mg/kg). At 1\u00a0day, 2\u2009days, and 3\u2009days following injection, the mice were euthanized via carbon dioxide inhalation and cervical dislocation, and their major organs were harvested and imaged by using a UVP iBOX Explorer Imaging Microscope (UVP) equipped with a 750\u2013780\u2009nm band\u2010pass excitation filter and an 800\u2009nm long\u2010pass emission filter. MFI for each organ was measured using the region of interest (ROI) function of ImageJ .Animal studies were approved and conducted according to the Institutional Animal Care and Use Committee of Brigham and Women's Hospital, Boston, MA. CLC\u2010IR800 was prepared by incubating CLC with IRdye 800CW NHS ester for 2\u00a0h at RT. The crude was purified by a desalting column . For ex vivo biodistribution studies, BALB/c mice were injected 4.13s.c. implanted with 1.0\u2009\u00d7\u2009105 4T1 cells. At 14\u2009days postimplantation, these mice received a single iv injection of CLC\u2010PTX* conjugate . After 1\u00a0day, the mice were euthanized via carbon dioxide inhalation and cervical dislocation, and their major organs were harvested and imaged by using a UVP iBOX Explorer Imaging Microscope (UVP) equipped with a 455\u2013495\u2009nm band\u2010pass excitation filter and a 513\u2013557\u2009nm band\u2010pass emission filter. MFI of the organs was measured following subtraction of autofluorescence, using the ROI function of ImageJ.CLC\u2010PTX* was prepared with the identical protocol for the synthesis of CLC\u2010PTX except the use of PTX* . BALB/c mice were 4.1418: 100\u2009\u00d7\u20092.1\u2009mm, 2.6\u00a0\u03bcm) under the following gradient elution system: 10% acetonitrile/water to 100% acetonitrile/water for 20\u2009min with a flow rate of 0.3\u00a0ml/min. Extracted ion is 854\u2009m/z.BALB/c mice were injected intravenously with PTX or CLC\u2010PTX at an equivalent PTX dose of 2.5\u00a0mg/kg. Sera from three mice were collected at 6\u00a0h and 2\u2009days following drug administration. The serum samples were stored at \u221220\u00b0C until analysis. Serum samples (200\u2009\u03bcl) were mixed with acetate buffer for 1\u00a0day at 37\u00b0C. PTX in the serum was extracted by incubation with acetonitrile (6\u00a0ml) for 2\u00a0h at 37\u00b0C. After centrifugation , the supernatants were analyzed by HPLC. High\u2010resolution electrospray ionization mass spectra were acquired using Agilent LC\u2010q\u2010TOF Mass Spectrometer 6530\u2010equipped with a 1290 uHPLC system. The collected samples were injected into a reversed\u2010phase HPLC column were 4.165 4T1 cells in the left fourth mammary gland. C57BL/6 mice were inoculated subcutaneously with 1.0\u2009\u00d7\u2009105 B16, LLC1, or Pan02 cells in the right rear flanks. When the tumor size reached ~100\u2009mm3, the mice were randomly divided into three groups (n\u00a0=\u00a06). All groups received treatments iv; the first group was injected with PBS (control), the second group with free PTX , and the final group with CLC\u2010PTX . The treatment schedule consisted of twice\u2010per\u2010week injections for 2\u2009weeks. The tumor size and body weight of the mice were monitored during the treatment course. The length (l) and width (w) of the tumor was measured by a digital Vernier caliper, and tumor volume (V) was defined as V=l\u00d7w2/2. We evaluated the TGI rate of each group (TGI%=Vc\u2212Vt/Vc\u2212Vi\u00d7100), in which Vc is the volume of the control tumor at the end of the study, Vt is the volume of drug\u2010treated tumor at the end of the study, and Vi is the volume of tumor at the initial treatment.BALB/c mice were implanted subcutaneously with 1.0\u2009\u00d7\u200910Before the diameter of the tumor reached ~2\u00a0cm, the mice were euthanized, and lungs, livers, kidneys, spleen, tumor, and draining inguinal lymph nodes were harvested and embedded in optimum cutting temperature (OCT) compound .4.17Frozen OCT blocks of tumors and LNs were cut using a cryostat (Leica) into 8\u2010\u03bcm thick sections and cells were incubated in glass\u2010bottomed well plate (Lab Tek\u2122). Meanwhile, cell samples were also stained by the following procedure. The samples were stained using anti\u2010pan\u2010cytokeratin , anti\u2010Melan\u2010A , anti\u2010LYVE\u20101 , anti\u2010fibronectin , anti\u2010CD31 , anti\u2010caspase\u20103 , anti\u2010mouse/human Ki\u201067 antibodies, or anti\u2010HSP70 . Dye\u2010conjugated secondary antibodies were used for the addition of the fluorescence signals. DAPI was used to stain the cell nuclei. The stained tissue sections were imaged using a fluorescent confocal microscope and an EVOS FL2 auto microscope or FluoView FV\u201010i Olympus Laser Point Scanning Confocal Microscope . After splitting each color channel, the fluorescent area for each color channel was measured with ImageJ . The expression level for each marker was calculated on the basis of normalization with the DAPI signal.4.18Major organs were immediately fixed in 4% paraformaldehyde, embedded in paraffin, and sliced into 5\u2010\u03bcm\u2010thick sections, which were then stained with H&E for histological examination. Organ histology was viewed and imaged under light microscopy (EVOS FL2).4.19iv injection of CLCs was measured by the Infinity Urea kit (Thermo Fisher Scientific) and compared against a standard BUN solution of 100\u2009mg/dl (Sigma\u2010Aldrich) using a VersaMax microplate reader , as per the protocol provided by Thermo Fisher Scientific. The creatinine (Cr) of the serum following iv injection of CLCs was measured using the microplate reader, according to the instructions provided in the Mouse Creatinine Assay Kit .BUN of the C57BL/6 mice serum following 4.20n) in the experiments was as follows: six for tumor growth analysis, six for lung metastatic foci assay, three for the fluorescent biodistribution, and three for the tissue immunostaining study. The fluorescence in vitro assay was independently conducted three times for statistical analysis. All data are expressed as the mean\u2009\u00b1\u2009SD from at least three independent samples or experiments. Differences between the two groups were analyzed by an unpaired Student's t\u2010test. Comparisons between multiple groups were determined using one or two\u2010way analysis of variance with Holm\u2010Sidak's post hoc test. A p value <0.05 was considered statistically significant .All statistical analysis was conducted using GraphPad Prism 7 software . The expression level of each marker in the immunostaining experiments was determined by dividing each DAPI level. The sample size ; data curation (lead); formal analysis (lead); investigation (lead); methodology (lead); validation (lead); visualization (lead). Liwei Jiang: Conceptualization ; data curation (lead); formal analysis (lead); investigation ; methodology . Jing Zhao: Data curation (supporting); investigation ; methodology . Leonard D. Shultz: Funding acquisition ; resources ; supervision . Dale L. Greiner: Methodology ; resources ; supervision . Munhyung Bae: Data curation (supporting); methodology (supporting). Xiaofei Li: Data curation (supporting); methodology (supporting). Farideh Ordikhani: Conceptualization (supporting); formal analysis (supporting); methodology (supporting). Rui Kuai: Supervision (supporting); validation (supporting). John Joseph: Data curation (supporting); formal analysis (supporting); methodology (supporting). Vivek Kasinath: Data curation (supporting); formal analysis (supporting); funding acquisition ; supervision (supporting); validation (supporting). David R. Elmaleh: Conceptualization ; formal analysis (supporting); methodology (supporting); resources (supporting); supervision (supporting). Reza Abdi: Conceptualization (lead); data curation (lead); formal analysis (lead); funding acquisition (lead); investigation (lead); methodology (lead); project administration (lead); resources (lead); software (lead); supervision (lead); validation (lead); visualization (lead).All authors declare no conflict of interests.https://publons.com/publon/10.1002/btm2.10273.The peer review history for this article is available at Appendix S1: Supporting informationClick here for additional data file."} +{"text": "These aluminates have been characterised by X\u2010ray crystallography and NMR spectroscopy. While the lithium aluminate forms a monomer, the heavier alkali metal aluminates exist as polymeric chains propagated by non\u2010covalent interactions between the alkali metal cations and the alkyldihydropyridyl units. Solvates [(THF)Li(tBuDHP)(TMP)Al(iBu)2] and [(TMEDA)Na(tBuDHP)(TMP)Al(iBu)2] have also been crystallographically characterised. Theoretical calculations show how the dispersion forces tend to increase on moving from Li to Rb, as opposed to the electrostatic forces of stabilization, which are orders of magnitude more significant. Having unique structural features, these bimetallic compounds can be considered as starting points for exploring unique reactivity trends as alkali\u2010metal\u2010aluminium hydride surrog[ATES].A series of group\u20051 hydrocarbon\u2010soluble donor free aluminates [AM( Saturated 2,2,6,6\u2010tetramethylpiperidide, TMP, has long played a key role in synthesis, here combined with an unsaturated dihydropyridine unit, 2\u2010tBuC5H5N, both contribute to the stability of a series of alkali metal aluminate structures by engaging in non\u2010covalent interactions with alkali metal cations, the extent of which depends on the alkali metal. Thermal Volatility Analysis (TVA) of the resultant samples showed that the propensity of hydride expulsion of the AM(tBuDHP) complexes increased down the group from lithium to potassium, following the trend of decomposition of saline hydrides.In 2015, our group reported another strategy, wherein a saturated C\u2212H bond in an alkyldihydropyridyl anion bound to lithium in has a track record in stabilizing a range of organolithium components to form heterobimetallic \u2018ate\u2019 complexes. These ates have had an important role to play in the development of bimetallic cooperativity.[iBu)2Al(TMP)] in tandem have ascertained that the sterically encumbered TMP ligands allow the lithium amide to display its basic function by deprotonation of an aryl/alkyl substrate, while the organoaluminium moiety traps the new, less hindered organolithium molecule in a method labelled as \u201ctrans\u2010metal\u2010trapping\u201d (TMT).[tBuDHP)], an amide carrying a surrogate hydride, as an alternative to Li(TMP). From a structural perspective this would open up a new avenue to isolate rarer aluminates of heavier alkali metals. Hence, our target here was to synthesise and characterise a new class of ate compound combining the complete set of alkali metal dihydropyridines with the dialkylamidoaluminium reagent [(iBu)2Al(TMP)], in effect heteroleptic alkali metal aluminium hydride surrog[ATES].Di\u2010iso\u2010butylaluminium\u20102,2,6,6\u2010tetramethylpiperidide [2Al(TMP)] was subjected to a suspensiontBuDHP)] in n\u2010pentane, the resultant mixture turned homogeneous at room temperature (Scheme\u20056D6 solution hinted at the formation of a new species as evident from a definite shift in the dihydropyridyl C\u2212H resonances in comparison to those in free Li(tBuDHP) . Meanwhile, a concentrated solution of the above mixture in n\u2010pentane cooled to \u221220\u2009\u00b0C overnight yielded colourless crystalline blocks. Single crystal X\u2010ray analysis of the crystals unveiled a solvent\u2010free monomer of a bimetallic ate complex, [Li(tBuDHP)(TMP)Al(iBu)2] (1) analogues ether, PMDETA=N,N,N\u2032,N\u2032\u2032,N\u2032\u2032\u2010pentamethyldiethylenetriamine, and Me6TREN=tris[2\u2010(dimethylamino)ethyl]amine].4H4)]n allows the alkali metal to modify its bonding nature towards the anionic N\u2010heterocyclic ligand from \u03c3\u2010bonding in [(PMDETA)Na(NC4H4)]2 to \u03c0\u2010bonding in the corresponding lower\u2010order and higher\u2010order zincate derivatives .1 fall in the range of 2.272(3)\u2005\u00c5 to 2.411(3)\u2005\u00c5. The sp3 hybridized ring carbon is slightly elevated from the conjugated plane lying at a distance of 2.705(3)\u2005\u00c5 from lithium. The alternating arrangement of short and long bond lengths within C6\u2212C7\u2212C8\u2212C9\u2212N1 \u2005\u00c5 in contrast to those in the solvated structures of Li(tBuDHP) reported earlier by our group .1 is related to some of the diamido\u2010dialkyl lithium aluminate examples in the literature1 is unusual being a structurally well\u2010defined example of a donor\u2010free monomeric ate featuring \u03c0\u2010stabilization.Pleasingly, it was found that when an equivalent amount of (1\u2009a) cannot be cast as a substitute to Li(TMP) since one decisive criterion for a TMT system states that the lithium and aluminium species should not co\u2010complex with each other to any significant extent to form an ate in order to display unique reactivity profiles. The lower steric profile of Li(tBuDHP) rules it out of consideration as a partner in trans\u2010metal\u2010trapping chemistry as it readily co\u2010complexes with the organoaluminium moiety. However, the formation of 1 and 1\u2009a can be treated as among the first illustrations of a trapped amido anion with a surrogate hydride as opposed to the trapped carbanions in all previous examples.It may be assumed that Li(tBuDHP) and having recently reported rubidium and caesium aluminyl compounds,tBuDHP). Owing to the practical difficulties involved in the process of isolating a pure batch of heavy alkali metal tert\u2010butoxide, in benzene provided the corresponding alkali metal alkoxides [Rb\u2212OtAm and Cs\u2212OtAm], obtained as white powders upon evacuation of the solvent and other volatiles under reduced pressure (Scheme\u2005tBuDHP) in benzene at room temperature allowed the precipitation of unsolvated hydride surrogates of rubidium [Rb(tBuDHP)] and caesium [Cs(tBuDHP] in quantitative yields. Each compound can be stored under inert conditions as a solid powder after washing away any lithium impurities with benzene. The identities of the above compounds were confirmed by multinuclear NMR spectroscopy (See Supporting Information for NMR characterisation). The dihydropyridyl signals in the 1H\u2005NMR spectra were in the range of 3.2\u2005ppm to 6.7\u2005ppm and comparable to those of the lighter congeners.tBuDHP) and Cs(tBuDHP) respectively in deuterated THF. Following the solubility trend of alkali metal dihydropyridines, Rb(tBuDHP) and Cs(tBuDHP) were found to be insoluble in hydrocarbon solvents. A possible explanation for this observation is that with increase in atomic radii down the group, the tendency to form higher order oligomers/polymers also intensifies.On the back of the progress made with Li2Al(TMP)] to combine with the heavier members of the group one tBuDHPs and remain homogeneous in solution (Scheme\u2005tBuDHP) and [(iBu)2Al(TMP)] produced colourless blocks of crystals after leaving the mixture with slow evaporation of hexane at room temperature. The structure of the compound was determined by single crystal X\u2010ray diffraction studies and identified as [Na(tBuDHP)(TMP)Al(iBu)2]\u221e contact of 2.977(2)\u2005\u00c5 from a neighbouring dihydropyridyl anion to construct a polymeric zig\u2010zag chain. The asymmetric unit of 2 bears a resemblance to 1 by virtue of having a similar bridging alignment [M\u2212N1\u2212Al\u2212N2] with a distorted tetrahedral geometry at the aluminium centre.Benzene was found to be a suitable medium for [Na(tBuDHP)(TMP)Al(iBu)2] (2\u2009a) were formed aluminate [(TMEDA)Na(TMP)(iBu)Al(iBu)2] protons were found to be split into three separate multiplets in the 1H NMR spectrum , of which two protons appear shielded at \u22120.02 (1H) and \u22120.30\u2005ppm (1H) respectively indicating the close interaction of the sodium centre to the axial H\u2010 atoms of the TMP moiety in the donor\u2010free aluminate. The structures of 2 and 2\u2009a bear significance on the grounds of the paucity of structures of bisamido bridged sodium aluminates with only a few crystallographically characterised examples present in the literature from the Fedyushkin and Chivers groups.[When an equivalent amount of TMEDA was added to a benzene solution of d Figure\u2005\u2010top. Inte Figure\u2005\u2010bottom. s groups.21biBu)Al(iBu)2] .[iBu)2Al(TMP)] induced the deprotonation of one \u03b1\u2010CH3 of a TMP unit to give the monomer [(TMEDA)K(\u03bc\u2010TMP*)(\u03bc\u2010iBu)Al(iBu)], where TMP* represents a CH3\u2010 and NH\u2010deprotonated dianionic variant of TMP.tBuDHP) produced the first donor free bis\u2010amido potassium aluminate in [K(tBuDHP)(TMP)Al(iBu)2]\u221e (3) in a 74\u2009% yield. Similarly, crystals from a Rb\u2212Al combination in benzene were obtained in a yield of 75\u2009% after removing the solvent in vacuo followed by adding hexane to the resultant yellow oil at room temperature. Aluminates 3, and [Rb(tBuDHP)(TMP)Al(iBu)2]\u221e (4) K(\u03bc\u2010amide), which lies opposite to the sp3 carbon (C5) atom [3.926\u2005\u00c5 in 2>3.319\u2005\u00c5 in 3>3.294\u2005\u00c5 in 4]. Contrary to the above observation, it is found that the distances between neighbouring methyl groups (C1) on the tBu substituent and the alkali metal follows a reverse order . While Li prefers to sit \u2018trans\u2019 to the tBu group of the tBuDHP, Na likes to rest itself on the site \u2018cis\u2019 to the tBu group of the tBuDHP ligand. This is evident from the contrasting CH3(TMP)\u22c5\u22c5\u22c5AM contacts Na(TMP)(iBu)Al(iBu)2}iBu)Al(iBu)2}].4 are distinctively shorter than the ones reported earlier by our group in the homoleptic TMEDA solvate [(TMEDA)Rb(TMP)]2.The \u03c0\u2010stabilization offered by the tBuDHP) and [(iBu)2Al(TMP)] in benzene solution could be proved from the comparable 1H\u2005NMR resonances against those of its lighter siblings, attempts to crystallize the same from its oil were unsuccessful. Addition of hexane to the oily substance resulted in the formation of a sticky solid. The presence of a variety of nitrogen and oxygen donors such as PMDETA, TMEDA, THF or 18\u2010crown\u20106\u2010ether in a hexane suspension of the bimetallic mixture failed to produce crystalline samples suitable for X\u2010ray crystallographic study. However, the solubility of the compound in benzene, points to the effectiveness of the aluminium agent in minimizing the oligomerization of the surrogate caesium hydride.While the formation of a co\u2010complexation product from a mixture of Cs. The electrostatic interaction energy was calculated according to Equation\u2005(1). We minimized basis set superposition error (BSSE) by calculating a counterpoise corrected interaction energy which increased electronic interaction energies of these four complexes by an average of 32\u2005kJ\u2009mol\u22121.It has been well documented that attractive London Dispersion Forces (LDF) influence the stability, structure, and reactivity of organometallic compounds.28b Late\u22121 for lithium to \u2212393\u2005kJ\u2009mol\u22121 for rubidium. As evident from Figure\u2005E) (kJ\u2009mol\u22121) in order of increasing stability=\u2212630.23 (Li)>\u2212511.65 (Na)>\u2212410.02 (K)>\u2212370.45 (Rb)], which is antithetical to the calculated LDF values [Energy (E) (kJ\u2009mol\u22121) in order of increasing stability=\u22124.47 (Li)<\u221215.07 (Na)<\u221218.92 (K)<\u221222.67 (Rb)] . These results support the existence of the observed short contacts [AM\u22c5\u22c5\u22c5H\u2212C(TMP)] in the donor\u2010free heavy alkali\u2010metal aluminates (3 and 4). Surprisingly, considering the high utility of TMP and the numerous structures it is found in,In order to better visualize and understand the myriad of non\u2010covalent interactions (NCI) we used the NCIPlot software with ultrafine integration to generate the representations in Figure\u20051\u20134 at 85\u2009\u00b0C in deuterated benzene. This showed that lithium aluminate 1 is the most stable, with no evidence of MH elimination/rearomatized pyridine formation even after heating for 24\u2005h. However, this study provided evidence of metal hydride elimination, as seen by the appearance of the characteristic 2\u2010t\u2010butylpyridine resonances in the 1H\u2005NMR spectra over 2\u20133\u2005days. In order to preliminarily test the surrogate hydride reactivity of these aluminates, experiments with the lightest and heaviest aluminates of the series (Li and Cs) were carried out at room temperature in hexane and benzene solution respectively with stoichiometric amounts of benzophenone. Quenching these mixtures with H2O resulted in the precipitation of insoluble hydroxide salts along with the formation of benzhydrol, which was characterised by NMR spectroscopy . However, these results do not explain the origin of the hydride source, since a previous report by our group indicated the availability of masked (\u03b2) hydride on the isobutyl groups bound to aluminium in the reduction of benzophenone, mimicking a Meerwein\u2010Pondorf\u2010Verley type mechanism.1 in C6D6 at room temperature . The appearance of characteristic isobutene resonances (\u03b4=4.73 and 1.59\u2005ppm) in the 1H\u2005NMR spectrum suggests the role of the masked hydride from the isobutyl groups. Addition of a further equivalent of benzophenone to this mixture revealed a complete loss of isobutyl resonances, confirming the aforementioned possibility. To our surprise, addition of a third equivalent of benzophenone at room temperature did not yield any 2\u2010t\u2010butylpyridine. The dihydropyridyl resonances appear shifted compared to both the starting material and also homometallic [Li(tBuDHP)], while the 7Li resonance shifts from \u22122.10 in 1 to 1.90\u2005ppm , suggesting an unidentified bimetallic species is formed. This lack of reactivity of the dihydropyridyl moiety suggests that the presence of the aluminium component provides a stabilizing influence since [Li(tBuDHP)] itself readily reduces benzophenone with concomitant generation of 2\u2010t\u2010butylpyridine. When this experiment was replicated with isolated crystals of the rubidium aluminate (4), the appearance of both isobutene as well as the 2\u2010t\u2010butylpyridine resonances in the 1H NMR spectrum were seen, hinting towards increased surrogate hydride reactivity of the heavier group one aluminates . Resonances at 6.09 and 6.27\u2005ppm in the 1H\u2005NMR spectrum allude to the fact that there are potentially two distinct \u2212OC(H)Ph2 environments in the product which has been witnessed previously in homometallic [(TMP)(Ph2(H)CO)Al(\u03bc\u2010OC(H)Ph2)]2 which shows distinct bridging and terminal resonances.We next attempted to study the stability of these complexes in light of the presence of these stabilizing interactions, by heating samples of the donor free ates tBuDHP)] into the chemistry of aluminates. The dialkyl amido aluminium compound [(iBu)2Al(TMP)] plays a fundamental role as a stabilizer by delivering the alkali metal surrogate hydride [AM(tBuDHP)] into a hydrocarbon solvent when added in stoichiometric quantities. The crystal structures of [Li(tBuDHP)(TMP)Al(iBu)2] (1), [Na(tBuDHP)(TMP)Al(iBu)2]\u221e (2), [K(tBuDHP)(TMP)Al(iBu)2]\u221e (3), and [Rb(tBuDHP)(TMP)Al(iBu)2]\u221e (4) demonstrated that the dihydropyridyl anion can act as a N\u2010heterocyclic group with a surrogate hydride providing stability to the alkali metals through \u03c0\u2010donation. These structures are one of the first in a series of donor free ates with two distinct amido bridges. The difference in the aggregation states is reflected in their solubility. While the lithium congener is soluble in an aliphatic hydrocarbon solvent, it takes an aromatic (benzene) solvent to dissolve the other group one aluminates. The polymers can be broken down into monomeric units by the employment of neutral Lewis base donors as observed during the isolation of [(THF)Li(tBuDHP)(TMP)Al(iBu)2] (1\u2009a) and [(TMEDA)Na(tBuDHP)(TMP)Al(iBu)2] (2\u2009a). Energy calculations on the stable donor free complexes provided us with an insight into the nature of alkali\u2010metal\u2010ligand interactions where a definite trend in increased contribution of attractive London Dispersion Forces can be observed on moving down group one, which is in sharp contrast to the electrostatic forces of attraction. The NCI plots demonstrated that the methylene groups of a saturated TMP ligand can lend stability to the large cations of group one. Preliminary test reactions suggest that the stability imparted on the dihydropyridyl unit as a consequence of the presence of the organoaluminium fragment modifies its reactivity as a metal hydride source, with the degree of modification depending upon the identity of the alkali metal. Our ongoing work is to investigate further this facet of their behaviour as well as to probe other reactivity scenarios in both stoichiometric and catalytic applications.In summary, this study has exported alkali metal dihydropyridines : \u03b4 0.98\u2005ppm , \u03b4 3.83\u2005ppm , \u03b4 4.94\u2005ppm , \u03b4 5.34\u2005ppm , \u03b4 6.27\u2005ppm , \u03b4 7.14\u2005ppm , \u03b4 1.72\u2005ppm , \u03b4 0.92\u2005ppm , \u03b4 0.80\u2005ppm , \u03b4 1.96\u2005ppm , \u03b4 1.47\u20131.54\u2005ppm , \u03b4 1.11\u20131.16\u2005ppm , \u03b4 2.13\u2005ppm , \u03b4 0.19\u20130.35\u2005ppm , \u03b4 0.49\u20130.57\u2005ppm ; 13C {1 H} NMR : \u03b4 147.62\u2005ppm (\u2212CH(6)[DHP]), \u03b4 123.42\u2005ppm (\u2212CH(4)[DHP]), \u03b4 107.16\u2005ppm (\u2212CH(3)[DHP]), \u03b4 99.09\u2005ppm (\u2212CH(5)[DHP]), \u03b4 59.70\u2005ppm (\u2212CH(2)[DHP]), \u03b4 24.76\u2005ppm (\u2212tBu[DHP]), \u03b4 40.68\u2005ppm (quaternary[DHP]), \u03b4 45.45+46.24\u2005ppm (\u03b2\u2010CH2[TMP]), \u03b4 17.81\u2005ppm (\u03b3CH2[TMP]), \u03b4 36.69+38.01\u2005ppm (CH3[TMP]), \u03b4 51.99+51.76\u2005ppm (2\u00d7quaternary[TMP]), \u03b4 26.73\u2005ppm (\u2212CH[iBu]), \u03b4 27.97+28.10\u2005ppm (\u2212CH3[iBu]), The resonances of \u2212CH2[iBu] could not be observed;7Li \u03b4=\u22121.71\u2005ppm (s).tBuDHP)(TMP)Al(iBu)2)] (1\u2009a)Synthesis of [(THF)Li(: Li(tBuDHP)iBu2AlTMPtBuDHP)(TMP)Al(iBu)]\u221e (2)Synthesis of [Na(: Na(tBuDHP)iBu2AlTMP26H50AlNaN2] (440.66\u2005g\u2009mol\u22121): C 70.87, H 11.44, N 6.36; Found: C 70.80, H 11.36, N 5.98. 1H\u2005NMR : \u03b4 1.18\u2005ppm , \u03b4 3.76\u2005ppm , \u03b4 4.59\u2005ppm , \u03b4 4.74\u2005ppm , \u03b4 5.85\u2005ppm , \u03b4 7.11\u2005ppm , \u03b4 1.35\u2005ppm , \u03b4 \u22120.30\u2005ppm , \u03b4 \u22120.02\u2005ppm , \u03b4 1.73\u2005ppm , \u03b4 1.09+1.15+1.39+1.51\u2005ppm , \u03b4 1.09\u2005ppm , \u03b4 1.43\u20131.47\u2005ppm , \u03b4 2.46\u2005ppm , \u03b4 0.38\u20130.87\u2005ppm ; 13C {1 H} NMR : \u03b4 147.65\u2005ppm (\u2212CH(6)[DHP]), \u03b4 125.41\u2005ppm (\u2212CH(4)[DHP]), \u03b4 105.02\u2005ppm (\u2212CH(3)[DHP]), \u03b4 94.96\u2005ppm (\u2212CH(5)[DHP]), \u03b4 59.96\u2005ppm (\u2212CH(2)[DHP]), \u03b4 25.66\u2005ppm (\u2212tBu[DHP]), \u03b4 41.55\u2005ppm (quaternary[DHP]), \u03b4 45.27\u2005ppm (\u03b2\u2010CH2[TMP]), \u03b4 17.72\u2005ppm (\u03b3CH2[TMP]), \u03b4 37.56+38.80\u2005ppm (CH3[TMP]), \u03b4 27.34+27.51\u2005ppm (\u2212CH[iBu]), \u03b4 28.24+28.59+29.27+29.94\u2005ppm (\u2212CH3[iBu]), The resonances of \u2212CH2[iBu] and quatenary[TMP] could not be observed.tBuDHP)(TMP)Al(iBu)] (2\u2009a)Synthesis of [(TMEDA)Na(: Na(tBuDHP)iBu2AlTMPtBuDHP)(TMP)Al(iBu)2] (2a) was not obtained, which may be attributed to decomposition during shipping and/or sample preparation. Best values are given, nevertheless. Elemental analysis: Calculated values for C32H66AlN4Na (556.86\u2005g\u2009mol\u22121): C 69.02, H 11.95, N 10.06; Found: C 68.98, H 11.73, N 7.59. 1H\u2005NMR : \u03b4 0.94\u2005ppm , \u03b4 3.74\u2005ppm , \u03b4 4.70\u2005ppm , \u03b4 4.76\u2005ppm , \u03b4 6.02\u2005ppm , \u03b4 7.04\u2005ppm , \u03b4 2.37\u2005ppm , \u03b4 2.27\u2005ppm , \u03b4 1.70\u2005ppm , \u03b4 1.31\u20131.41\u2005ppm , \u03b4 1.04\u2005ppm , \u03b4 2.06\u2005ppm , \u03b4 0.04\u20130.47\u2005ppm ; 13C {1 H} NMR : \u03b4 147.06\u2005ppm (\u2212CH(6)[DHP]), \u03b4 123.98\u2005ppm (\u2212CH(4)[DHP]), \u03b4 105.62\u2005ppm (\u2212CH(3)[DHP]), \u03b4 93.75\u2005ppm (\u2212CH(5)[DHP]), \u03b4 60.21\u2005ppm (\u2212CH(2)[DHP]), \u03b4 25.19\u2005ppm (\u2212(CH3)3[DHP]), \u03b4 41.32\u2005ppm (quaternary[DHP]), \u03b4 57.91\u2005ppm (\u2212CH2[TMEDA]), \u03b4 46.73\u2005ppm (\u2212CH3[TMEDA]), \u03b4 45.83\u2005ppm (\u03b2\u2010CH2[TMP]), \u03b4 18.16\u2005ppm (\u03b3CH2[TMP]), \u03b4 26.09\u2005ppm (CH3[TMP]), \u03b4 51.65\u2005ppm (quaternary[TMP]), \u03b4 26.92+27.03\u2005ppm (\u2212CH[iBu]), \u03b4 27.97+28.60+29.06\u2005ppm (\u2212CH3[iBu]), The resonances of \u2212CH2[iBu] could not be observed.tBuDHP)(TMP)Al(iBu)]\u221e (3)Synthesis of (3) was not obtained, which may be attributed to decomposition during shipping and/or sample preparation. Best values are given, nevertheless. Elemental analysis: Calculated values for [C26H50AlKN2] (456.77\u2005g\u2009mol\u22121): C 68.37, H 11.03, N 6.13; Found: C 67.84, H 9.98, N 4.67. 1H\u2005NMR : \u03b4 1.25\u2005ppm 3[DHP]+6H, \u2212CH3[TMP]+2H, \u03b2\u2010CH2[TMP]), \u03b4 3.79\u2005ppm , \u03b4 4.54\u2005ppm , \u03b4 4.67\u2005ppm , \u03b4 5.74\u2005ppm , \u03b4 7.04\u2005ppm , \u03b4 1.38\u2005ppm , \u03b4 0.87\u2005ppm , \u03b4 1.47\u2005ppm , \u03b4 2.50\u2005ppm , \u03b4 0.41\u20130.83\u2005ppm ; 13C {1 H} NMR : \u03b4 147.92\u2005ppm (\u2212CH(6)[DHP]), \u03b4 125.67\u2005ppm (\u2212CH(4)[DHP]), \u03b4 104.02\u2005ppm (\u2212CH(3)[DHP]), \u03b4 95.18\u2005ppm (\u2212CH(5)[DHP]), \u03b4 66.28\u2005ppm (\u2212CH(2)[DHP]), \u03b4 26.08\u2005ppm (\u2212tBu[DHP]), \u03b4 41.78\u2005ppm (quaternary[DHP]), \u03b4 18.13\u2005ppm (\u03b3CH2[TMP]), \u03b4 51.50\u2005ppm (quaternary[TMP]), \u03b4 27.44+27.73\u2005ppm (\u2212CH[iBu]), \u03b4 28.44+28.76+29.63+29.88\u2005ppm (\u2212CH3[iBu]). The resonances of \u2212CH2[iBu] could not be observed. The resonances of \u03b2\u2010CH2[TMP] and \u2212CH3[TMP] were not assigned due to overlap of the methyl signals (from TMP and iBu groups) in 1H13C\u2010HSQC NMR spectrum.iBu)]\u221e (4)Synthesis of (4) was not obtained, which may be attributed to decomposition during shipping and/or sample preparation. Best values are given, nevertheless. Elemental analysis: Calculated values for [C26H50AlRbN2] (503.14\u2005g\u2009mol\u22121): C 62.07, H 10.02, N 5.57; Found: C 61.75, H 9.73, N 4.62. 1H\u2005NMR : \u03b4 1.30\u2005ppm , \u03b4 3.85\u2005ppm , \u03b4 4.56\u2005ppm , \u03b4 4.61\u2005ppm , \u03b4 5.66\u2005ppm , \u03b4 7.03\u2005ppm , \u03b4 0.83\u2005ppm , \u03b4 1.63\u2005ppm , \u03b4 1.34\u2005ppm , \u03b4 1.48\u2005ppm , \u03b4 2.50\u2005ppm , \u03b4 0.43\u20130.75\u2005ppm ; 13C {1 H} NMR : \u03b4 148.13\u2005ppm (\u2212CH(6)[DHP]), \u03b4 125.87\u2005ppm (\u2212CH(4)[DHP]), \u03b4 103.95\u2005ppm (\u2212CH(3)[DHP]), \u03b4 94.94\u2005ppm (\u2212CH(5)[DHP]), \u03b4 60.59\u2005ppm (\u2212CH(2)[DHP]), \u03b4 26.30\u2005ppm (\u2212tBu[DHP]), \u03b4 41.88\u2005ppm (quaternary[DHP]), \u03b4 43.66\u2005ppm (\u03b2\u2010CH2[TMP]), \u03b4 18.33\u2005ppm (\u03b3CH2[TMP]), \u03b4 33.69+34.08\u2005ppm (CH3[TMP]), \u03b4 51.55\u2005ppm (quaternary[TMP]), \u03b4 27.41+27.75\u2005ppm (\u2212CH[iBu]), \u03b4 28.58+29.09+29.68+29.76\u2005ppm (\u2212CH3[iBu]), The resonances of \u2212CH2[iBu] and quaternary[TMP] could not be observed.The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "In the crystal structure, centrosymmetric mol\u00adecules are linked through inter\u00admolecular N\u2014H\u22efO hydrogen bonds into sheets extending parallel to (0 2(en)2)] , has been synthesized. The complex mol\u00adecules are located on inversion centers. Two DNBA anions monodentately coordinate the CdII atom through an oxygen atom of the carboxyl\u00adate group while two en mol\u00adecules coordinate in a chelate fashion, resulting in a distorted O2N4 coordination set. There is a weak intra\u00admolecular hydrogen bond of 3.099\u2005(4)\u2005\u00c5 between the non-coordinating oxygen atom of the carboxyl\u00adate group and one of the en amine groups. Three relatively weak inter\u00admolecular N\u2014H\u22efO hydrogen bonds associate complex mol\u00adecules into sheets extending parallel to and H\u22efH (21.1%) inter\u00adactions.During systematic investigations of bioavailability and biological action enhancement of well known compounds with low bioactivity, a new mixed-ligand metal complex, [Cd(DNBA) It is a well known bidentate chelating ligand for coordination complexes to 102.66\u2005(12)\u00b0, indicating a rather strong distortion from the ideal octa\u00adhedral shape. The conformation of the complex mol\u00adecule is stabilized through a weak intra\u00admolecular hydrogen bond [3.099\u2005(4)\u2005\u00c5 and 143\u2005(3)\u00b0] between the N4\u2014H4A donor and the O2 acceptor (Table\u00a01S(6). Most coplanar with the aromatic ring is the N1O2 nitro group [dihedral angle of 3.873\u2005(3)\u00b0] while the carboxyl\u00adate group is considerably twisted from the aromatic ring [dihedral angle = 19.332\u2005(9)\u00b0]. The arrangement of the N2O2 nitro group is inter\u00admediate between the latter two, the corresponding dihedral angle being 13.529\u2005(6)\u00b0.In the crystal of the title compound, the complex mol\u00adecules are located on inversion centers. Two symmetry-related DNBA anions monodentately coordinate to Cdms Fig.\u00a01. The bonr Table\u00a01 definingA\u22efO4i and N4\u2014H4B\u22efO4ii hydrogen bonds define rings with graph-set notation via N3\u2014H3B\u22efO5iii hydrogen bonds, forming sheets extending parallel to (0Cg1\u22efCg1 = 3.715\u2005(3)\u2005\u00c5, slippage = 1.608\u2005\u00c5, symmetry operation: 1\u00a0\u2212\u00a0x, \u2212y, 1\u00a0\u2212\u00a0z; Cg1 is the centroid of the phenyl (C1\u2013C6) ring].There are three relatively weak inter\u00admolecular hydrogen bonds in the crystal structure Table\u00a01. N4\u2014H4A\u22ef1) Fig.\u00a02. The sheCrystal Explorer 17.5 analysis was carried out using rm Fig.\u00a03 shows thet al., 20162(en)2)] complex. The silver complexes with refcodes EQOKEA 2 was slowly added an ethanol solution (4\u2005ml) containing en (60 \u03bcl) and DNBA under constant stirring. A colourless crystalline product was obtained at room temperature by slow solvent evaporation after 6\u2005d. Single crystals for X-ray structure determination were selected from this product. Yield: 65%. Elemental analysis for C18H22CdN8O12 (654.83): calculated C 33.02; H 3.39; N 17.11%; found: C 32.96; H 3.32; N 17.08%.To an aqueous solution (2.5\u2005ml) of Cd I. DOI: 10.1107/S2414314620008433/wm4132Isup2.hklStructure factors: contains datablock(s) I. DOI: 2011747CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-022-13747-4, published online 11 June 2022Correction to: The original version of this\u00a0Article contained an error in the y-axis\u00a0label of\u00a0Figure\u00a04(C), where \u201cstereoacuity improvement (%)\u201d should read \u201cstereoacuity improvement\u201d.The original Figure\u00a0The original Article has been corrected."} +{"text": "In the crystal, corrugated layers parallel to the ab plane are formed by a combination of C\u2014H\u22efO, C\u2014H\u22ef\u03c0(ring) and \u03c0-stacking inter\u00adactions.The inner part of the ester substituent is nearly perpendicular to the di\u00adhydro\u00adpyridazine ring while the 17H20N2O4, the inner part of the ester substituent is nearly perpendicular to the di\u00adhydro\u00adpyridazine ring, forming a dihedral angle of 83.21\u2005(7)\u00b0. In the crystal, inversion dimers are formed by pairwise C\u2014H\u22efO inter\u00adactions with the dimers connected into chains extending along the b-axis direction by C\u2014H\u22ef\u03c0(ring) inter\u00adactions. The chains are connected by \u03c0-stacking inter\u00adactions to give corrugated layers parallel to the ab plane. The terminal ethyl group is disordered over two two sets of sites with the major component having a site occupancy factor of 0.715\u2005(10)In the title mol\u00adecule, C This latter angle indicates that the inner end of the substituent on N2 is nearly perpendicular to the tetra\u00adhydro\u00adpyridazine ring. The C2\u2014C3\u2014C5\u2014C6 torsion angle of \u22129.4\u2005(2)\u00b0 indicates that the centroid of the 4-meth\u00adoxy\u00adphenyl ring is only slightly below the plane of the pyridazine ring. This conformation appears to be the result of the inter\u00admolecular \u03c0-stacking inter\u00adaction (see below).B\u22efO1 inter\u00adactions (Table\u00a01b-axis direction by C16\u2014H16B\u22efCg1 inter\u00adactions (Table\u00a01i\u2013C11i rings [symmetry code: (i) \u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a0ab plane -one pre\u00adcursor. To this pyridazine derivative (0.05\u2005mol) was added potassium carbonate (0.1\u2005mmol), tetra\u00adbutyl\u00adammonium bromide (0.01\u2005mmol) and 2-ethyl bromo\u00adacetate (0.1\u2005mol) in di\u00admethyl\u00adformamide (20\u2005ml). The mixture was stirred for 24\u2005h at room temperature. At the end of the reaction, the solution was filtered and the solvent evaporated under reduced pressure. The residue was washed with water and methyl\u00adene chloride. The solvent was removed and colourless blocks of the title compound were obtained by recrystallization of the product from its acetone solution.A mixture of 3-(4-meth\u00adoxy\u00adbenzyl\u00adidene)-4-oxo\u00adpenta\u00adnoic acid (0.05\u2005mol) and hydrazine hydrate (0.1\u2005mol) in ethanol (100\u2005ml) was refluxed for 2\u2005h. The precipitate that formed was filtered off and recrystallized from acetone solution to obtain the 5-(4-meth\u00adoxy\u00adbenz\u00adyl)-6-methyl\u00adpyridazin-3(2\u22121): 1743 , 1660 (C=ON), 1599 (C=C), 1205 (C\u2014N), 1011 and 1145 . 1H NMR (p.p.m.): 1.23 ; 2.22 ; 2.33 ; 3.85 ; 4.17 ; 4.87 ; 6.48 ; 6.93\u20136.96 ; 7.25\u20137.27 . 13C NMR (p.p.m.): 14.11 (CH3); 21.03 ; 25.21 ; 37.67 (CH2); 51.34 (CH2); 60.95 (CH2); 127.13\u2013127.44 (CH aromatic); 129.13\u2013130.35 (CH aromatic); 132.12 (C\u2014C\u03b1 aromatic); 136.51 ; 138.49 ; 144.97 ; 147.17 (C=N); 161.19 ; 169.52 .Yield 79%; m.p. 406\u2013408\u2005K. IR global, I. DOI: 10.1107/S241431462200582X/tk4077Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S241431462200582X/tk4077Isup3.cmlSupporting information file. DOI: 2175897CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II ion has an N4Cl2 octa\u00adhedral coordination sphere defined by three N atoms of the tridentate 2,3,5,6-tetra-2-pyridyl\u00adpyrazine ligand, one N atom of the pyridine ligand and two Cl\u2212 anions.The central Ni 2(C5H5N)(C24H16N6)], the NiII ion is six-coordinated in a distorted octa\u00adhedral coordination environment defined by three N atoms of the tridentate 2,3,5,6-tetra-2-pyridyl\u00adpyrazine ligand, one N atom of the pyridine ligand and two Cl\u2212 anions, with the latter being mutually trans. The complex is disposed about a twofold rotation axis along the a axis. The complex molecules are connected in the crystal via C\u2014H\u22efCl, C\u2014H\u22efN and \u03c0\u2013\u03c0 [closest inter-centroid separation = 3.7446\u2005(14)\u2005\u00c5 between pyridyl rings].In the title complex, [NiCl The Ni\u2014N[pyrazine\u00ad(N1) or pyrid\u00adyl] bond lengths are roughly equivalent, with distances of 2.008\u2005(3) \u2013 2.1026\u2005(19)\u2005\u00c5. The pyrazine ring (N1\u2014C1i) slightly deviates from planarity, with a maximum deviation of 0.057\u2005(2)\u2005\u00c5 for the C2 atom from the least-squares plane of the ring. The dihedral angles between the nearly planar pyridyl rings and the least-squares plane of their carrier pyrazine ring are 14.90\u2005(4)\u00b0 for the coordinating pyridyl ring (N3\u2014C7) and 54.42\u2005(9)\u00b0 for the non-coord\u00adinating pyridyl ring (N4\u2014C12), respectively. The dihedral angle between the pyrazine ring and the pyridine ligand (N5\u2014C13i) is 57.8\u2005(1)\u00b0.In the title complex, the central Nins Fig.\u00a01. The comCg1 (the centroid of the ring N3/C3\u2013C7) and Cg1ii [symmetry code: (ii) x, x\u00a0\u2212\u00a0y, \u2212z\u00a0+\u00a0In the crystal, the complex displays numerous inter- and intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between adjacent six-membered rings. The most significant inter\u00adaction of this kind is that between s Table\u00a01 that cons Table\u00a01.2\u00b76H2O in ethanol (20\u2005ml) was added 2,3,5,6-tetra-2-pyridyl\u00adpyrazine , followed by stirring for 24\u2005h at rooom temperature. The formed precipitate was separated by filtration, washed with ethanol and acetone, and dried at 323\u2005K, to give a brown powder (0.5045\u2005g). Brown crystals suitable for X-ray analysis were obtained by slow evaporation from its pyridine/N,N-di\u00admethyl\u00adformamide (DMF) solution at 333\u2005K.To a solution of NiClCrystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621000948/wm4144sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314621000948/wm4144Isup2.hklStructure factors: contains datablock(s) I. DOI: 2058988CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Yarrowia\u2009lipolytica as an alternative host for ergothioneine production. We expressed the biosynthetic enzymes EGT1 from Neurospora\u2009crassa and EGT2 from Claviceps\u2009purpurea to obtain 158\u2009mg\u00b7L\u22121 of ergothioneine in small\u2010scale cultivation, with an additional copy of each gene improving the titer to 205\u2009mg\u00b7L\u22121. The effect of phosphate limitation on ergothioneine production was studied, and finally, a phosphate\u2010limited fed\u2010batch fermentation in 1\u2009L bioreactors yielded 1.63\u2009\u00b1\u20090.04\u2009g\u00b7L\u22121 ergothioneine in 220\u2009h, corresponding to an overall volumetric productivity of 7.41\u2009mg\u00b7L\u22121\u00b7h\u22121, showing that Y.\u2009lipolytica is a promising host for ergothioneine production.Ergothioneine is a naturally occurring antioxidant that has shown potential in ameliorating neurodegenerative and cardiovascular diseases. In this study, we investigated the potential of the Crabtree\u2010negative, oleaginous yeast Yarrowia\u2009lipolytica is a viable host to produce the antioxidant ergothioneine. Yarrowia\u2009lipolytica could reach ergothioneine production levels comparable to or exceeding those reported in other hosts, such as Escherichia\u2009coli and Saccharomyces\u2009cerevisiae, by genomic integration of the ergothioneine biosynthetic pathway and fed\u2010batch fermentation on mineral medium.Here, we report that the oleaginous yeast CDW, cell dry weightERG, ergothioneineMES, 2\u2010(N\u2010morpholino)ethanesulfonic acidSAM, S\u2010adenosylmethionineThe amino acid\u2010derived nutraceutical ergothioneine (ERG) has recently gained much scientific interest for its potential application in preventing or treating neurodegenerative and cardiovascular diseases , 2. CurrS\u2010adenosylmethionine was bought from Sigma\u2010Aldrich . Synthetic genes were ordered through the GeneArt Gene Synthesis service of Thermo Fisher Scientific or the custom gene synthesis service of IDT . Sequencing results were obtained through Eurofins Genomics using their Mix2Seq kit.The n ST6512 was used\u22121 of glucose, 7.5\u2009g\u00b7L\u22121 of (NH4)2SO4, 14.4\u2009g\u00b7L\u22121 of KH2PO4, 0.5\u2009g\u00b7L\u22121 of MgSO4.7H2O, 2\u2009mL\u00b7L\u22121 of trace metal solution, and 1\u2009mL\u00b7L\u22121 of vitamin solution, adjusted to pH 6.0 using 2\u2009m of NaOH. Mineral medium without phosphate was prepared using the same recipe, but KH2PO4 was substituted with the same concentration of 2\u2010(N\u2010morpholino)ethanesulfonic acid (MES) hydrate. High glucose mineral medium without phosphate consisted of 50\u2009g\u00b7L\u22121 of glucose, 20\u2009g\u00b7L\u22121 of (NH4)2SO4, 30\u2009g\u00b7L\u22121 of MES hydrate, 1.25\u2009g\u00b7L\u22121 of MgSO4.7H2O, 5\u2009mL\u00b7L\u22121 of trace metal solution, and 2.5\u2009mL\u00b7L\u22121 of vitamin solution, adjusted to pH 6.0 using 2\u2009m of NaOH.Mineral medium consisted of 20\u2009g\u00b7L\u22121 of CaCl2.2H2O, 4.5\u2009g\u00b7L\u22121 of ZnSO4.7H2O, 3\u2009g\u00b7L\u22121 of FeSO4.7H2O, 1\u2009g\u00b7L\u22121 of H3BO3, 1\u2009g\u00b7L\u22121 of MnCl2.4H2O, 0.4\u2009g\u00b7L\u22121 of Na2MoO4.2H2O, 0.3\u2009g\u00b7L\u22121 of CoCl2.6H2O, 0.1\u2009g\u00b7L\u22121 of CuSO4.5H2O, 0.1\u2009g\u00b7L\u22121 of KI, and 15\u2009g\u00b7L\u22121 of EDTA. Vitamin solution consisted of 50\u2009mg\u00b7L\u22121 of biotin, 200\u2009mg\u00b7L\u22121 of p\u2010aminobenzoic acid, 1\u2009g\u00b7L\u22121 of nicotinic acid, 1\u2009g\u00b7L\u22121 of calcium pantothenate, 1\u2009g\u00b7L\u22121 of pyridoxine HCl, 1\u2009g\u00b7L\u22121 of thiamine HCl, and 25\u2009g\u00b7L\u22121 of myo\u2010inositol.Trace metal solution consisted of 4.5\u2009g\u00b7LY.\u2009lipolytica was performed using the EasyCloneYALI method . , 9. Y.\u2009lY.\u2009lipolytica. Phosphate limitation was a viable strategy to limit biomass accumulation in the bioreactors. ERG titer reached 1.63\u2009\u00b1\u20090.04\u2009g\u00b7L\u22121 after 220\u2009h of fermentation in mineral medium with glucose as the only carbon source.In conclusion, we integrated the ERG biosynthesis pathway into oleaginous yeast SH, DBK, and IB are named inventors on a European Patent application covering parts of the work described above.IB and DBK conceived the study; IB and JLM supervised the study; SAH, IHJ, and JLM designed the experiments; SAH, IHJ, and MR performed the experiments and data analysis; SAH and IB wrote the manuscript; SAH, IB, DBK, IHJ, and JLM reviewed and revised the manuscript; IB and JLM provided resources for the study; IB acquired funding for the study.Fig.\u2009S1. Bacterial ergothioneine biosynthesis pathway.Fig.\u2009S2. The effect of phosphate limitation on the growth of the ergothioneine\u2010producing Yarrowia lipolytica strain ST10264.Fig.\u2009S3. Ergothioneine production and biomass accumulation of ST10264 under phosphate\u2010limited fed\u2010batch conditions in a single 1 L bioreactor with an initial amount of 240\u2009mg KH2PO4.Table\u2009S1. List with genes and their DNA sequences used in this paper.Table\u2009S2. List of primers used in this study for cloning purposes.Table\u2009S3. List of primers used in this study for sequencing.Table\u2009S4. List of biobricks used in this study.Table\u2009S5. List of plasmids used in this study, made by USER cloning.Table\u2009S6. List of strains used in this study.Click here for additional data file."} +{"text": "The crystal structure of the \u2018magic\u2019 mushroom natural product baeocystin is reported for the first time. N-methyl\u00adtryptamine {systematic name: 3-[2-(methylazaniumyl)ethyl]-1H-indol-4-yl hydrogen phosphate}, C11H15N2O4P, has a single zwitterionic mol\u00adecule in the asymmetric unit. The mol\u00adecule has an intra\u00admolecular N\u2014H\u22efO hydrogen bond between the ammonium cation and the hydro\u00adphosphate anion. In the crystal, the mol\u00adecules are linked by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds into a three-dimensional network.The title compound, baeocystin or 4-phosphor\u00adyloxy- This minor tryptamine natural product was first isolated from the Psilocybe baeocystis mushroom in 1968 , the metabolite of baeocystin, as a Psilocybe natural product in mice, which is strongly correlated with 5-HT2A receptor-mediated psychoactive effects \u00b0. The phosphor\u00adyloxy group is similarly turned away from the indole plane, with a C5\u2014C6\u2014O1\u2014P1 torsion angle of 33.8\u2005(3)\u00b0. Both groups are turned to the same side of the indole ring, which is likely supported by an intra\u00admolecular N2\u2014H2A\u22efO4 hydrogen bond distance of 0.87\u2005(1)\u2005\u00c5, N\u2014H(ammonium) distances of 0.90\u2005(1)\u2005\u00c5, and an O\u2014H distance of 0.90\u2005(1)\u2005\u00c5. Isotropic displacement parameters were set to 1.2 Ueq of the parent nitro\u00adgen atoms and 1.5 Ueq of the parent oxygen atom. All other hydrogen atoms were placed in calculated positions . Isotropic displacement parameters were set to 1.2 Ueq of the parent carbon atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022004467/hb8018sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022004467/hb8018Isup2.hklStructure factors: contains datablock(s) I. DOI: 2169087CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound crystallizes with two mol\u00adecules in the asymmetric unit. In the crystal, asymmetric, bifurcated N\u2014H\u22ef hydrogen bonds link the mol\u00adecules into [100] chains. The packing is consolidated by weak C\u2014H\u22efO inter\u00adactions. 25H25N3O4, crystallizes with two mol\u00adecules in the asymmetric unit. In both mol\u00adecules, the central ethane hydrazide moiety is almost planar. In the crystal, asymmetric, bifurcated N\u2014H\u22ef hydrogen bonds link the mol\u00adecules into [100] chains. The packing is consolidated by weak C\u2014H\u22efO inter\u00adactions.The title compound, C This latter ring and the C1A toluyl ring are almost perpendicular, with a dihedral angle of 82.1\u2005(1)\u00b0The title compound crystallizes with two mol\u00adecules, B, the central ethane hydrazide moiety N2B/N3B/C17B/O3B is also close to planar, with an r.m.s. deviation of 0.01\u2005\u00c5 for the fitted atoms. The equivalent dihedral angles to those in mol\u00adecule A are 13.3\u2005(2), 80.5\u2005(2) and 86.1\u2005(1)\u00b0, respectively.For mol\u00adecule A and B mol\u00adecules into a zigzag chain propagating in the a-axis direction, as shown in Fig.\u00a02In the crystal, bifurcated, asymmetric N\u2014H\u22ef hydrogen bonds Table\u00a01 link altet al., 2004o-tolyl groups at the other end of the mol\u00adecule: in the current structure these are 82.1\u2005(1) and 86.1\u2005(1)\u00b0 while in JUFCIY the equivalent angles are 46.98\u2005(5) and 48.23\u2005(4)\u00b0.A search for related structures resulted in two close matches. The first is the parent azide from which the current structure is derived (Chopra E)-2-(Meth\u00adoxy\u00adimino)-2-{2-[(2-methyl\u00adphen\u00adoxy)meth\u00adyl]phen\u00adyl}ethano\u00adhydrazide was refluxed for 8\u2005h with p-meth\u00adoxy\u00adbenzaldehyde in 20\u2005ml of absolute ethanol with the addition of 5 drops of glacial acetic acid to obtain a white-colored product. This was dissolved in DMSO and, by the process of slow evaporation, colourless needles of the title compound grew. I. DOI: 10.1107/S2414314620010603/hb4356Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314620010603/hb4356Isup3.cmlSupporting information file. DOI: 2020760CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title hydro\u00adchloride salt, the cation shows an unusually long Si\u2014C bond length. In the crystal, the cations and anions are linked by N\u2014H\u22efCl hydrogen bonds to generate [001] chains. N-{[Diphen\u00adyl(vin\u00adyl)sil\u00adyl]meth\u00adyl}-2-methyl\u00adpropan-2-amine, C19H25NSi, is a newly synthesized secondary amino\u00admethyl\u00adsilane that can be used, for example, to study carboli\u00adthia\u00adtion reactions of vinyl\u00adsilanes. Because the neutral compound did not crystallize well, the hydro\u00adchloride salt, C19H26NSi+\u00b7Cl\u2212, was formed, in which the two chloride ions in the asymmetric unit have crystallographic + species. In the crystal, the cations and anions are linked by N\u2014H\u22efCl hydrogen bonds to generate [001] chains. To further investigate the inter\u00admolecular inter\u00adactions, a Hirshfeld surface analysis was performed, which showed that H\u22efH, C\u22efH/H\u22efC and H\u22efCl/Cl\u22efH contacts contribute 70.4, 20.0 and 8.3%, respectively. Because the (amino\u00admeth\u00adyl)silane 1 did not crystallize well, the hydro\u00adchloride salt 2 was formed to characterize the compound via X-ray diffraction. For example, the newly synthesized (amino\u00admeth\u00adyl)vinyl\u00adsilane 1, C19H25NSi, can be used for investigations of a carboli\u00adthia\u00adtion reaction of the silane\u2019s vinyl group via lithiumalkyls. The received product can be used for the synthesis of functionalized alcohols by a Tamao oxidation silanes known to date because the synthesis is not feasible due to the high energy requirement and reaction time. With the assistance of a 2.2 crystallized in a few minutes from an aqueous 1\u2005M HCl solution of 1 at room temperature as a hydro\u00adchloride salt, C19H26NSi+\u00b7Cl\u2212, in the form of colorless needles in the centrosymmetric space group P21/n. The mol\u00adecular structure is illustrated in Fig.\u00a01Compound 3sp\u2014SiX3 compounds \u00b0, which is caused by the steric repulsion of the aromatic hydrogen atoms. The Si1\u2014C1 bond length is 1.8577\u2005(11)\u2005\u00c5 and C1\u2014C2 is 1.3293\u2005(16)\u2005\u00c5; the latter is positioned at the end of the default range of Csp2\u2014Csp2 bonds, which lie between 1.299 and 1.328\u2005\u00c5.The Si1\u2014C15 bond length in the cation is 1.9117\u2005(10)\u2005\u00c5, which is slightly longer than the average for an Si\u2014C bond and the Si1\u2014C15\u2014N1 bond angle is 116.21\u2005(7)\u00b0. The Si\u2014C bond lengths are normally in the range of 1.857 to 1.905\u2005\u00c5 for CA\u2014N1\u2014H1B), the angles between the C atoms and the H atoms are 107.8\u2005(10)\u00b0 (H1B\u2014N1\u2014C15) and 109.6\u2005(9)\u00b0 (H1A\u2014N1\u2014C15). Between the carbon atoms, the angle is 117.10\u2005(7)\u00b0 (C15\u2014N1\u2014C16). All angles vary slightly from the ideal tetra\u00adhedron angles of 109.5\u00b0: the large C\u2014N\u2014C angle results from the bigger space requirement of the carbon atoms in comparison to the H atoms. The sum of angles around the nitro\u00adgen atom is 441.7\u00b0, so the overall structure is distorted tetra\u00adhedral. The bond length between N1 and C15 is 1.4928\u2005(12)\u2005\u00c5 and it is 1.5330\u2005(13)\u2005\u00c5 between N1 and C16. In the literature, C3sp\u2014N bond lengths are in the range of 1.4816 to 1.5034\u2005\u00c5, so the N1\u2014C16 bond is slightly extended.The cationic nitro\u00adgen center features a slightly disordered tetra\u00adhedral geometry. The angle between the hydrogen atoms is 107.2\u2005(13)\u00b0 \u2005\u00c5. This may be due to the different surroundings of the Cl1 and Cl2 ions in the crystal. As shown in Fig.\u00a03para-hydrogen atoms H6 with a C6\u22efCl1 distance of 4.0013\u2005(11)\u2005\u00c5 while Cl2 accepts two weak, near linear hydrogen-bond contacts [C7\u2014H7\u22efCl2: 165.74\u2005(7)\u00b0] from the aromatic meta-hydrogen atoms H7 with a C7\u22efCl2 distance of 3.9419\u2005(12)\u2005\u00c5. Both contacts are formed by the same aromatic ring. The bond angle for N1\u2014H1B\u22efCl2 is 164.0\u2005(13)\u00b0, compared to 172.8\u2005(13)\u00b0 for N1\u2014H1A\u22efCl1. They differ from the optimal angle of 180\u00b0 because of the different surroundings in the crystal packing.In the extended structure of 2 Fig.\u00a02, the cat2 Fig.\u00a02 to generdnorm in the range from \u22120.54 to 1.49 arbitrary units, generated by CrystalExplorer2021 and D12(3), which means that the hydrogen bond extends from N1\u2014H1A\u22efCl1 to another H1A\u2014N1 grouping of a neighboring mol\u00adecule. H2 can also be assigned D11(2) and D12(3) -2-meth\u00adoxy\u00admethyl-1-[1-phenyl\u00adeth\u00adyl(dimeth\u00adyl)sil\u00adyl\u00admeth\u00adyl]pyrrolidinium iodide, C17H30NOSi+\u00b7I\u2212 \u2005\u00c5] and the Si\u2014C\u2014N bond angle is comparable [115.86\u2005(14)\u00b0]. The lengths between the carbon atoms and the cationic nitro\u00adgen center are similar to the corresponding bond lengths in 2 [1.498\u2005(3) and 1.494\u2005(3)\u2005\u00c5].In LAGLUE, the N\u2014H\u22efCl hydrogen bond has a slightly longer N\u22efCl separation (3.124\u2005\u00c5) than compound 2 but the Si\u2014C\u2014N bond angle is similar [114.9\u2005(2)\u00b0] and the C\u2014N bond is a bit extended [1.505\u2005(6)\u2005\u00c5]. This could be caused by the ring strain of the aziridine ring and the electron-withdrawing effect of the (nitro\u00adphen\u00adyl)sulfonyl group located at the nitro\u00adgen center. In addition, the Si\u2014Csp2 bond length is 1.859\u2005(7)\u2005\u00c5, which is only slightly longer that the value for 2.In WOLSEY, the Si\u2014C distance of 1.871\u2005(4)\u2005\u00c5 is shorter than in Finally, in AGILIL, the Si\u2014C bond length is slightly shorter [1.907\u2005(7)\u2005\u00c5] and the Si\u2014C\u2014N bond angle is slightly extended [120.8\u2005(4)\u00b0], which is caused by the cyclic structure of the compound. The C\u2014N distance is equal [1.498\u2005(8)\u2005\u00c5] and the cyclic N\u2014C bond lengths marginally shorter [1.509\u2005(8) and 1.516\u2005(8)\u2005\u00c5], again due to the cyclic structure.+) to 2. In WAVXAW and DAFKUT, the Si\u2014C bond lengths are 1.9074\u2005(11) and 1.907\u2005(3)\u2005\u00c5, respectively, comparable to the value in 2. These extended bond lengths are due to the same electronic effects already described.The structures of WAVXAW and DAFKUT contain a similar structure motive was added to N-{[diphen\u00adyl(vin\u00adyl)sil\u00adyl]meth\u00adyl}-2-methyl\u00adpropan-2-amine (1) at room temperature. The product (2) was formed after five minutes as colorless needles.The reaction scheme for the synthesis of compound 6.A and H1B were positioned geometrically (C\u2014H = 0.95\u20131.00\u2005\u00c5) and refined using a riding model, with Uiso(H) = 1.2Ueq(C) for CH2 and CH hydrogen atoms and Uiso(H) = 1.5Ueq(C) for CH3 hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022009112/hb8033sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022009112/hb8033Isup2.hklStructure factors: contains datablock(s) I. DOI: 2207007CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "IV\u2014S bond lengths. The packing of the complexes enable inter\u00admolecular inter\u00adactions between the qdt ligands and an amine hydrogen atom (hydrogen bonding) or other qdt ligands (\u03c0\u2013\u03c0 stacking).The title compound exhibits typical Pt 8H4N2S2)2(C5H6N2)2]\u00b7C2H6OS or trans-[Pt(4-ap)2(qdt)2]\u00b7dmso the centrosymmetric complex exhibits Pt\u2014S distances in agreement with other PtIV\u2014S bond lengths found in platinum(IV) di\u00adthiol\u00adene complexes. The qdt ligands have inter\u00admolecular inter\u00adactions with an amine hydrogen atom on a 4-ap ligand (hydrogen bonding) and have sandwich \u03c0\u2013\u03c0 inter\u00adactions with a neighboring qdt ligand.In the structure of the title solvated complex, [Pt(C These distances are similar to those in other platinum(IV) complexes containing bis\u00ad(di\u00adthiol\u00adene) ligands and either trans-bis\u00ad(NH3) co-ligands, with Pt\u2014S distances of 2.3434\u2005(8) and 2.3461\u2005(7)\u2005\u00c5 co-ligands, with a Pt\u2014S distance of 2.3619\u2005(8)\u2005\u00c5 2(mnt)2] complex and S2 . This tilt enables sandwich packing between inter\u00admolecular qdt ligands with a distance between centroids of the two qdt rings of 3.610\u2005\u00c5 atom on a neighboring qdt ligand, with a distance of 2.23\u2005\u00c5 (Table\u00a01The chelating qdt ligands of this platinum(IV) complex are slightly canted relative to the platinum-sulfur atoms, with a 15.59\u2005(11)\u00b0 angle between the plane of all the non-H atoms of the qdt ligand \u2005\u00c5 Fig.\u00a02, within \u00c5 Table\u00a01. N\u2014H\u22efO ha3 with 25\u2005ml of water and heating at 333\u2005K for 5\u2005h. Upon cooling to room temperature, the orange solution was added, via cannula, to a Schlenk flask containing 34.3\u2005mg of [Pt(4-ap)4](BF4)2, prepared in a similar manner to [Pt(pyz)4](BF4)2 2(qdt)]). Oxidation of platinum(II) to platinum(IV) likely occurred upon prolonged air exposure of the compound in solution I. DOI: 10.1107/S2414314622001018/wm4160Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622001018/wm4160Isup3.molSupporting information file. DOI: 2145270CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title piperidine derivative, the mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions into dimers related by twofold symmetry. 20H21NO, the dihedral angle between the phenyl ring is 47.5\u2005(1)\u00b0 and the piperidine ring adopts a chair conformation. In the crystal, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions into dimers with the mol\u00adecules related by twofold symmetry.In the title compound, C This leads to dimers with the mol\u00adecules related by twofold rotation symmetry and the C6\u2013C11 and C15\u2013C20 benzene rings are 70.31\u2005(11) and 79.00\u2005(11)\u00b0, respectively. The dihedral angle between the C6\u2013C11 and C15\u2013C20 benzene rings is 47.51\u2005(12)\u00b0. In the crystal, a C2\u2014H2\u22efry Fig.\u00a02. The N1\u2014A mixture of hexene-2-one (0.05\u2005mol), benzaldehyde (0.1\u2005mol), ammonium acetate (0.05\u2005mol) and ethanol (40\u2005ml) was heated gently and poured into ether (50\u2005ml) and treated with concentrated hydro\u00adchloric acid (25\u2005ml). The precipitated hydro\u00adchloride was washed with an ethanol\u2013ether mixture. The base was liberated by adding strong ammonia until the hydro\u00adchloride dissolved. Dilution with water afforded the free base. The pure compound was was further recrystallized with benzene\u2013petroleum ether to yield the title compound.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S241431462000526X/hb4345sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S241431462000526X/hb4345Isup2.hklStructure factors: contains datablock(s) I. DOI: 1996936CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II centre is coordinated by four pyrrole N atoms of the porphyrin ring and one S atom of the axial 4-methyl\u00adbenzene\u00adthiol\u00adate ligand. Two tetra\u00adhydro\u00adfuran mol\u00adecules and a potassium cation chelated inside a [2.2.2]cryptand are also present.The Mn 18H36N2O6)][Mn(C44H24Cl4N4)(C7H7S)]\u00b72C4H8O, were obtained by the solvent evaporation method. The MnII cation is coordinated by four pyrrole N atoms (Np) of the porphyrin ring and one S atom of the apical 4-methyl\u00adbenzene\u00adthiol\u00adate ligand with the average Mn\u2014Np and the apical Mn\u2014S bond lengths being 2.160\u2005(9) and 2.4642\u2005(8)\u2005\u00c5, respectively. Two tetra\u00adhydro\u00adfuran solvent mol\u00adecules and a potassium cation chelated inside a [2.2.2]cryptand are also present.Single crystals of the title compound, [K(C II porphyrin analogues are often used as substitutes and supplements for FeIII porphyrin model compounds when simulating the relevant reactions of heme-active sites, so as to provide a more comprehensive understanding of structure\u2013function relationships in hemoproteins [Mn(TPP)(1-MeIm)] has a large metal out-of-plane distance (\u039424 and \u03944 \u2265 0.51), and the authors suggested that when the dy2x2\u2013 orbital is populated, the metal atom is too far out of the porphyrin plane to permit effective inter\u00adaction with a sixth ligand. Subsequently, some other crystal structures of five-coordinate manganese porphyrins were reported, e.g. [K(222)][Mn(TPP)(CN)] ] (p-CH3PhS\u2212)].MnII porphyrinate, i.e. the [MnII(TpClPP)(p-CH3PhS\u2212)]\u2212 anion, is a [K(222)]+ cation in which the potassium ion is chelated inside a [2.2.2]cryptand mol\u00adecule. Six O and two N atoms of the [2.2.2]cryptand bind to the potassium cation, and the average K\u2014O and K\u2014N distances are 2.85\u2005(4) and 2.989\u2005(11)\u2005\u00c5, respectively. There are also two tetra\u00adhydro\u00adfuran (THF) solvent mol\u00adecules in the asymmetric unit, one of which is disordered.The mol\u00adecular entities of the title compound are shown in Fig.\u00a01II has a larger ionic radius than FeIII , longer M\u2014Np (Np is a porphyrin N atom) bond lengths [2.160\u2005(9)\u2005\u00c5 versus 2.057\u2005(5)\u20132.06\u2005(12)\u2005\u00c5], and a longer Mn\u2014S apical distance [2.4642\u2005(8)\u2005\u00c5 versus 2.298\u2005(5)\u20132.311\u2005(3)\u2005\u00c5] Cl], and [Mn(TpClPP)OH] were prepared according to a literature method ] was prepared by reduction of [MnIII(TpClPP)OH] with ethane\u00adthiol (1.5\u2005ml) in benzene (5\u2005ml) atmosphere using standard Schlenk techniques, and all solvents used were treated under anhydrous and anaerobic conditions. Benzene and tetra\u00adhydro\u00adfuran were distilled over sodium/benzo\u00adphenone. Hexane was distilled from sodium/potassium alloy under argon. Pyrrole, Synthesis of ([2.2.2]cryptand)potassium (4-methyl\u00adbenzene\u00adthiol\u00adato)mang\u00adanese(II) tetra\u00adhydro\u00adfuran disolvateII(TpClPP)] powder was dried under vacuum and then dissolved in 5\u2005ml of tetra\u00adhydro\u00adfuran. 6.0\u2005mg (0.050\u2005mmol) of p-CH3PhSK and 17.0\u2005mg (0.045\u2005mmol) of Kryptofix 222 were added to the solution. The mixture was stirred for about 30 minutes and hexane was layered. After 2 weeks, X-ray quality dark-purple flake-shaped crystals of [K(222)][Mn(TpClPP)(p-CH3PhS\u2212)] were obtained and a large well-formed specimen was selected for the diffraction experiment.The purple [MnA, C71A and C72A were restrained with the RIGU instruction to get a better disorder model. The refined occupancy ratio of the disordered atoms is 0.576\u2005(7):0.524\u2005(7).Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2414314622002413/bt4121sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622002413/bt4121Isup3.hklStructure factors: contains datablock(s) I. DOI: 2142607CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-benzoyl\u00adamido\u00adphosphate is reported and discussed.The crystal structure of the caesium salt of dimethyl- N-benzoyl\u00adamido\u00adphosphate, namely, aqua\u00ad[di\u00admeth\u00adyl (N-benzoyl\u00adamido-\u03baO)phospho\u00adnato-\u03baO]caesium, [Cs(C9H11NO4P)(H2O)] or CsL\u00b7H2O, is reported. The compound crystallizes in the monoclinic crystal system in the P21/c space group and forms a mono-periodic polymeric structure due to the bridging function of the dimethyl-N-benzoyl\u00adamido\u00adphosphate anions towards the caesium cations.The caesium salt of dimethyl- In addition, the phosphoryl or the carbonyl oxygen atom or both usually bridge the cations. Caesium salts of CAPhs have not been reported to date and are of inter\u00adest as possible dopants in oxide film materials for the improvement of their electric and electron functional characteristics . The oxygen atoms of the carbonyl and phosphoryl groups of the dimethyl-N-benzoyl\u00adamido\u00adphosphate anions act as \u03bc2-bridges between Cs+ cations , and each Cs+ cation links four ligand anions. Additionally, a water mol\u00adecule acts as a \u03bc2-bridge between two Cs+ cations.The asymmetric unit contains the Csle Fig.\u00a02a. The ons Fig.\u00a01. Additions Fig.\u00a02b, and e+ ion contacts nine oxygen atoms. It is involved in the six-membered Cs1\u2013O1\u2013C1\u2013N1\u2013P1\u2013O2 ring with one ligand by bonding with the oxygen atoms of the carbonyl and phosphoryl groups, in the four-membered Cs1\u2013O2\u2013P1\u2013O4 ring with another CAPh ligand by bonding with the oxygen atoms of the phosphoryl and meth\u00adoxy groups and in the six-membered Cs1\u2013O1\u2013C1\u2013N1\u2013P1\u2013O3 ring with the third ligand by bonding with the \u03bc2-oxygen atoms of the carbonyl and meth\u00adoxy groups. In addition, the Cs+ ion contacts with the \u03bc2-O3 atom of the fourth neighboring CAPh as well as with two mol\u00adecules of water , so the majority of the Cs\u2014O contacts might be considered as a mainly ionic type of bond. The Cs1\u2014O1 distance is the longest LH = 1.219\u2005(6)\u2005\u00c5, d(P\u2014O)LH = 1.461\u2005(4)\u2005\u00c5] and the C\u2014N and P\u2014N bond lengths are decreased , 1591 [\u03bd(CC)], 1535 [\u03bd(CO)], 1378 [\u03bd(CN)], 1205 [\u03bd(PO)], 1076, 1038, 928 [\u03bd(PN)], 838, 800, 734, 710, 540, 502,466, 452\u2005cm\u22121. The low-frequency shift of \u03bd(P=O) and \u03bd(C=O) bands in the IR spectrum of CsL\u00b7H2O with respect to HL(\u0394\u03bdLH(P=O) \u223c37cm\u22121, \u0394\u03bdLH(C=O) \u223c147cm\u22121] is typical for bidentate coordination of dimethyl-N-benzoyl\u00adamido\u00adphosphate. 1H NMR (DMSO-d6): \u03b4 = 3.24 , 3.54 , 7.27 , 8.04 . 31P NMR (acetone): \u03b4 = 15.2 (s).Cs7.Uiso(H) = 1.2\u20131.5Ueq(C). O-bound H atoms were refined with the restraints O5\u2014H5A = O5\u2014H5B = 0.84\u00b10.01\u2005\u00c5 and H5A\u22efH5B = 1.62\u00b10.02\u2005\u00c5 with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022012166/mw2194sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989022012166/mw2194Isup2.hklStructure factors: contains datablock(s) I. DOI: 2232690CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Iclaprim and mesylate mol\u00adecules are linked into a hydrogen-bonded mol\u00adecular tape, the central section of which is composed of fused rings. 2H-1-benzo\u00adpyran-5-yl)meth\u00adyl]pyrimidin-1-ium methane\u00adsulfonate, C19H23N4O3+\u00b7CH3O3S\u2212, is a salt made up from a protonated iclaprim mol\u00adecule and a mesylate anion. The pyrimidine and chromene units of the iclaprim mol\u00adecule form an orthogonal arrangement [inter\u00adplanar angle of 89.67\u2005(6)\u00b0], and the 3-nitro\u00adgen position of the pyrimidine ring is protonated. Four distinct N\u2014H\u22efO inter\u00adactions and an additional N\u2014H\u22efN hydrogen bond connect iclaprim and mesylate mol\u00adecules to one another, resulting in an infinite hydrogen-bonded mol\u00adecular tape structure. The central section of the tape is formed by a sequence of fused hydrogen-bonded rings involving four distinct ring types.The title compound, 2,6-di\u00adamino-5-[(2-cyclo\u00adpropyl-7,8-dimeth\u00adoxy- Iclaprim was synthesized according to the original route described by Jaeger al. 2005, using 32.3SO3\u2212 anion and an iclaprim cation in which the 3-nitro\u00adgen atom of the pyrimidine ring is protonated, i.e. N1 with the fused benzene ring of the chromene unit . With regard to the two bridging bonds, the torsion angles C3\u2014C4\u2014C5\u2014C6 [\u2013160.8\u2005(2)\u00b0] and C4\u2014C5\u2014C6\u2014C7 [\u201396.5\u2005(3)\u00b0] indicate that the C5\u2014C6 bond is twisted slightly out of the pyrimidine plane, whilst the C4\u2014C5 bond is oriented approximately perpendicular to the benzene ring. Accordingly, the two six-membered rings linked via C5 form an orthogonal arrangement with an inter\u00adplanar angle of 89.67\u2005(6)\u00b0. In the chromene moiety, the 7-meth\u00adoxy substituent is significantly twisted out of the ring plane [C10\u2014C9\u2014O3\u2014C19 = \u221270.3\u2005(3)\u00b0], whilst the 8-meth\u00adoxy substituent is almost coplanar with the plane of the fused benzene ring [C9\u2014C8\u2014O2\u2014C18 = 167.6\u2005(2)\u00b0]. The 2H-pyran ring displays the expected bond lengths [C12\u2014C13 = 1.323\u2005(4)\u2005\u00c5]. The program PLATON \u00b0, \u03c6 = 328.4\u2005(7)\u00b0 and q = 0.253\u2005(3)\u2005\u00c5, are consistent with the presence of a skew-boat conformation N1 Fig.\u00a01. The mol3.2 groups attached to the pyrimidine ring and the protonated N1 atom of the pyrimidine ring as potential hydrogen-bond donor groups. These hydrogen-bond donor functions are engaged in five distinct inter\u00admolecular N\u2014H\u22efA inter\u00adactions .Iclaprim mesylate was prepared according to a modified procedure based on the original synthesis by Jaeger al. 2005 shown in6.a axis. The refined value of the minor twin component fraction was 0.260\u2005(1). All H atoms were identified in difference-Fourier maps and those of NH and NH2 groups were refined with a restrained N\u2014H distance of 0.88\u2005(2)\u2005\u00c5 and their Uiso parameters refined freely. The H atoms at the cyclo\u00adpropyl ring were refined with a restrained C\u2014H distance of 0.96\u2005(2)\u2005\u00c5 and with Uiso(H) = 1.2Ueq(C). Other H atoms bonded to secondary CH2 (C\u2014H = 0.98\u2005\u00c5) or aromatic CH (C\u2014H = 0.94\u2005\u00c5) carbon atoms were positioned geometrically. Their Uiso parameters were set to 1.2Ueq(C). Methyl H atoms were idealized and included as rigid groups allowed to rotate but not tip (C\u2014H = 0.97\u2005\u00c5) and their Uiso parameters were set to 1.5 Ueq(C) of the parent carbon atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022011689/wm5666sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022011689/wm5666Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022011689/wm5666Isup3.cmlSupporting information file. DOI: 2224639CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-butyl-2,3-bis\u00ad(di\u00adcyclo\u00adhexyl\u00adamino)\u00adcyclo\u00adpropenimine is reported.The structure of the acid chloride salt of the superbase N-Butyl-2,3-bis\u00ad(di\u00adcyclo\u00adhexyl\u00adamino)\u00adcyclo\u00adpropenimine (1) crystallizes from benzene and hexa\u00adnes in the presence of HCl as a mono\u00adbenzene solvate of the hydro\u00adchloride salt, [1H]Cl\u00b7C6H6 or C31H54N3+\u00b7Cl\u2212\u00b7C6H6, in the P21/n space group. The protonation of 1 results in the generation of an aromatic structure based upon the delocalization of the cyclo\u00adpropene double bond around the cyclo\u00adpropene ring, giving three inter\u00admediate C\u2014C bond lengths of \u223c1.41\u2005\u00c5, and the delocalization of the imine-type C\u2014N double bond, giving three inter\u00admediate C\u2014N bond lengths of \u223c1.32\u2005\u00c5. Ion\u2013ion and ion\u2013benzene packing inter\u00adactions are described and illustrated. Originally reported as four-electron Lewis donors \u00adcyclo\u00adpropenimine (1) is a newer version of superbase with improved basicity, which has been explored as a catalyst for ring-opening polymerization. Cyclo\u00adpropenimines have a conjugate acid apK of about 27, an improvement over that of the superbase 2-tert-butyl-1,1,3,3-tetra\u00admethyl\u00adguanidine (BTMG), which has a apK of 23.56 \u2005\u00c5 on average. The C\u2014N bond to the protonated butyl nitro\u00adgen is only slightly shorter at 1.319\u2005(2)\u2005\u00c5. The three cyclo\u00adpropene C\u2014C bonds exhibit lengths consistent with aromaticity; the unique C\u2014C bond opposite the n-butyl group is 1.388\u2005(2)\u2005\u00c5, while the other two C\u2014C bonds are similar or slightly shorter at 1.377\u2005(2) and 1.383\u2005(2)\u2005\u00c5. Though these latter two bonds are equivalent under mol\u00adecular point symmetry, their differences are attributed to the asymmetric crystal packing environment of the P21/n space group. The comparable nature of the bond metrics of the three C\u2014N bonds and the three cyclo\u00adpropenyl C\u2014C bonds is consistent with aromatization, and an analogous aromatization of the C3N3 core of 1-mesityl-2,3-bis\u00ad(diiso\u00adpropyl\u00adamino)\u00adcyclo\u00adpropeniminium tetra\u00adfluoro\u00adborate was observed in the crystal structure of this salt , which ranges from about 6\u201325\u2005kJ\u2005mol\u22121 per \u03c0 electron , and 2.7169\u2005(6)\u2005\u00c5 from an intra\u00admolecular axial cyclo\u00adhexyl proton, H29A (+0.050). The crystal packing demonstrates that the C3N3 planes of all mol\u00adecules pack parallel to each other (as required by the space-group symmetry), with a normal slightly oblique to the (101) plane \u2005\u00c5, Fig.\u00a03n-butyl chain Cl\u00b7C6H6 were observed inside. The yield of crystalline [1H]Cl\u00b7C6H6 was significantly improved by the addition of HCl. To a glass shell vial containing 7.2\u2005mg of N-butyl-2,3-bis\u00ad(di\u00adcyclo\u00adhexyl\u00adamino)\u00adcyclo\u00adpropenimine, 2\u2005mL of benzene were added. A drop of dilute HCl (0.730 M) was added. This was diffused with 3\u2005mL of hexa\u00adnes in the outer vial for 2\u20133 days. Crystallization works best when the drop is not in contact with the walls of the vial where the crystals grow. Crystals were isolated by deca\u00adnting the liquid from the inner vial using a disposable pipette, and taking care to remove the visible aqueous HCl droplet with the first pipette draw. After removing the mother liquor, the crystals were rinsed with hexa\u00adnes. Yield 6.3\u2005mg (70%). Yields in this small-scale preparation ranged from 22% to 70% across multiple attempts. 1H NMR (ppm) 400\u2005MHz, CDCl3): \u03b4(ppm): 0.97 , 1.62\u20131.82 , 1.62\u20131.76 , 1.80 , 1.96 , 3.34 , 3.56 , 7.4 . 13C NMR (ppm) : \u03b4(ppm): 13.97, 19.94, 24.58, 25.84, 32.34, 33.79, 46.15, 59.55, 114.01, 128.35. FTIR (cm\u22121): 2926 (m), 2851 (m), 1503 (s), 1445 (m), 1383 (w), 1374 (w), 1345 (w), 1324 (w), 1253 (w), 1188 (w), 1180 (w), 1102 (w), 1092 (w), 1004 (w), 895 (w), 696 (m). Analysis calculated for C31H53N3\u00b70.5 C6H6 (%): C, 76.31; H, 10.38; N, 7.22. Found: C, 75.873; H, 10.83; N, 7.24. M.p. 353\u2013356\u2005K (decomposes).Initially, crystals of Cl\u00b7C6H6 using ORCA I. DOI: 10.1107/S2056989022008076/jq2013Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022008076/jq2013Isup3.cdxSupporting information file. DOI: 2200141CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Anopheles gambiae sensu lato (s.l.) mosquitoes to pyrethroids and their expression of metabolic enzymes.Malaria remains one of the most devastating diseases globally, and the control of mosquitoes as the vector is mainly dependent on chemical insecticides. Elevated temperatures associated with future warmer climates could affect mosquitoes' metabolic enzyme expression and increase insecticide resistance, making vector control difficult. Understanding how mosquito rearing temperatures influence their susceptibility to insecticide and expression of metabolic enzymes could aid in the development of novel tools and strategies to control mosquitoes in a future warmer climate. This study evaluated the effects of temperature on the susceptibility of Anopheles gambiae s.l. eggs obtained from laboratory-established colonies were reared under eight temperature regimes . Upon adult emergence, 3- to 5-day-old female non-blood-fed mosquitoes were used for susceptibility tests following the World Health Organization (WHO) bioassay protocol. Batches of 20\u201325 mosquitoes from each temperature regime (25\u201334\u00a0\u00b0C) were exposed to two pyrethroid insecticides (0.75% permethrin and 0.05% deltamethrin). In addition, the levels of four metabolic enzymes were examined in mosquitoes that were not exposed and those that were exposed to pyrethroids.An. gambiae s.l. mosquitoes exposed to deltamethrin and permethrin decreased at temperatures above 28\u00a0\u00b0C. In addition, mosquitoes reared at higher temperatures were more resistant and had more elevated enzyme levels than those raised at low temperatures. Overall, mosquitoes that survived after being exposed to pyrethroids had higher levels of metabolic enzymes than those that were not exposed to pyrethroids.Mortality in An. gambiae s.l. mosquitoes' susceptibility to pyrethroids and increased the expression of metabolic enzymes. This evidence suggests that elevated temperatures projected in a future warmer climate could increase mosquitoes' resistance to insecticides and complicate malaria vector control measures. This study therefore provides vital information, and suggests useful areas of future research, on the effects of temperature variability on mosquitoes that could guide vector control measures in a future warmer climate.This study provides evidence that elevated temperatures decreased The online version contains supplementary material available at 10.1186/s13071-022-05273-z. Anopheles gambiae could affect malaria control measures. The An. gambiae complex consists of nine morphologically identical sibling species. They include An. gambiae sensu stricto (s.s.), An. arabiensis, An. merus, An. melas, An. quadriannulatus, An. amharicus, An. bwambae, An. coluzzii, and An. fontenillei ) were first used to identify species of the protocol . Subsequectively . The PCRAn. gambiae s.l. mosquitoes raised at 25, 28, 30, 32, and 34\u00a0\u00b0C. Permethrin and deltamethrin insecticides were selected because they are the most common insecticides used in malaria control programs compared to those reared at 34\u00a0\u00b0C [1.94\u2009\u00d7\u200910\u20139 ], 30\u00a0\u00b0C [1.49\u2009\u00d7\u200910\u20139 ], 28\u00a0\u00b0C [8.85\u2009\u00d7\u200910\u201310 ], and 25\u00a0\u00b0C [7.21\u2009\u00d7\u200910\u201310 mol cytochrome P450/min/mg protein] with increasing temperature. Dunn's multiple range test showed that all but 30 vs 28\u00a0\u00b0C (P\u2009=\u20090.113) and 34 vs 30\u00a0\u00b0C (P\u2009=\u20090.010) showed a statistically significant difference in oxidase level mol cytochrome P450/min/mg protein] levels compared to those reared at 32\u00a0\u00b0C [2.40\u2009\u00d7\u200910\u20139 ], 25\u00a0\u00b0C [2.07\u2009\u00d7\u200910\u20139 ], and 28\u00a0\u00b0C [1.34\u2009\u00d7\u200910\u20139 mol cytochrome P450/min/mg protein] in MFO levels among the different temperature regimes. Further tests using Dunn's multiple range test showed a significant difference (P\u2009<\u20090.008) in the various temperature regime comparisons , 28\u00a0\u00b0C , and 30\u00a0\u00b0C and not exposed to pyrethroids were significantly lower than those that were exposed to pyrethroids. However, mosquitoes reared at 32\u00a0\u00b0C and not exposed to pyrethroids had significantly higher MFO levels than those exposed to pyrethroids in the GST levels in mosquitoes among the different temperature regimes. However, further analysis using Dunn's multiple range test showed statistically significant differences for 32 vs 25\u00a0\u00b0C (P\u2009<\u20090.001), 34 vs 25\u00a0\u00b0C (P\u2009=\u20090.002), 32 vs 28\u00a0\u00b0C (P\u2009<\u20090.001), 32 vs 30\u00a0\u00b0C (P\u2009<\u20090.001), and 34 vs 32\u00a0\u00b0C (P\u2009<\u20090.001) similar to that of the MFO level ], followed by 25\u00a0\u00b0C [2.72\u2009\u00d7\u200910\u20133 ], 30\u00a0\u00b0C [2.62\u2009\u00d7\u200910\u20133 ] and the lowest recorded at 28\u00a0\u00b0C [1.35\u2009\u00d7\u200910\u20133 mol cDNB/min/mg protein] in the GST level in mosquitoes among the different temperature regimes. Further statistical tests using Dunn's multiple range test showed statistically significant differences for 28 vs 25\u00a0\u00b0C (P\u2009=\u20090.002), 32 vs 28\u00a0\u00b0C (P\u2009<\u20090.001), and 32 vs 30\u00a0\u00b0C (P\u2009=\u20090.005) showed a statistically significant difference in GST levels ], followed by 34\u00a0\u00b0C [4.04\u2009\u00d7\u200910\u20137 ], 28\u00a0\u00b0C [2.83\u2009\u00d7\u200910\u20137 ], 25\u00a0\u00b0C [2.52\u2009\u00d7\u200910\u20137 ] and 30\u00a0\u00b0C [2.12\u2009\u00d7\u200910\u20137 mol \u03b1-naphthol/min/mg protein] in \u03b1-esterase level in mosquitoes among the different temperature regimes. Further statistical tests using Dunn's multiple range test showed a statistically significant difference for 32 vs 25\u00a0\u00b0C (P\u2009<\u20090.001), 34 vs 25\u00a0\u00b0C (P\u2009<\u20090.001), 32 vs 28\u00a0\u00b0C (P\u2009<\u20090.001), 32 vs 30\u00a0\u00b0C (P\u2009<\u20090.001), 34 vs 30\u00a0\u00b0C (P\u2009<\u20090.001), and 34 vs 32\u00a0\u00b0C (P\u2009<\u20090.001) ] and lowest at 28\u00a0\u00b0C [1.44\u2009\u00d7\u200910\u20137 (2.40\u2009\u00d7\u200910\u20137) mol \u03b1-naphthol/min/mg protein] in \u03b1-esterase levels in mosquitoes among the different temperature regimes. According to Dunn's multiple range tests, a statistical difference existed only for 30 vs 28\u00a0\u00b0C (P\u2009=\u20090.003) and 32 vs 28\u00a0\u00b0C (P\u2009=\u20090.001) showed a statistically significant difference in \u03b1-esterase level ]\u2009>\u200932\u00a0\u00b0C [2.87\u2009\u00d7\u200910\u20137 ]\u2009>\u200928\u00a0\u00b0C [1.38\u2009\u00d7\u200910\u20137 ]\u2009>\u200925\u00a0\u00b0C [1.36\u2009\u00d7\u200910\u20137 ]\u2009>\u200930\u00a0\u00b0C [1.30\u2009\u00d7\u200910\u20137 mol \u03b2-naphthol/min/mg protein] in \u03b2-esterase levels in mosquitoes with increasing temperature. However, post hoc analysis using Dunn's multiple range test showed a statistically significant difference for 32 vs 25\u00a0\u00b0C (P\u2009=\u20090.001), 34 vs 25\u00a0\u00b0C (P\u2009<\u20090.001), 34 vs 28\u00a0\u00b0C (P\u2009<\u20090.001), 34 vs 30\u00a0\u00b0C (P\u2009<\u20090.001), and 34 vs 32\u00a0\u00b0C (P\u2009=\u20090.003) at 28\u00a0\u00b0C to 2.64\u2009\u00d7\u200910\u20137 (1.63\u2009\u00d7\u200910\u20137) mol \u03b2-naphthol/min/mg protein at 32\u00a0\u00b0C in \u03b2-esterase levels in mosquitoes raised at different temperatures. Multiple comparisons using Dunn's test showed statistically significant differences for 28 vs 25\u00a0\u00b0C (P\u2009<\u20090.001), 30 vs 28\u00a0\u00b0C (P\u2009<\u20090.001), 32 vs 28\u00a0\u00b0C (P\u2009<\u20090.001), and 32 vs 30\u00a0\u00b0C (P\u2009=\u20090.007) in \u03b2-esterase levels to pyrethroids decreased with increasing temperature. Another important finding was that deltamethrin induced higher mortality in mosquitoes than permethrin, indicating that mosquitoes were resistant to permethrin based on WHO criteria [An. gambiae s.l. mosquitoes were more susceptible to deltamethrin than permethrin. With projections of warmer temperatures in coming years, especially in sub-Saharan Africa, high resistance to pyrethroids with increasing temperatures could affect the effectiveness of malaria control programs using pyrethroids [Mosquito susceptibility to pyrethroids decreased with increasing temperature. This may be attributed to the higher expression of enzymes at high temperatures . At highcriteria . These fcriteria . This cocriteria , which sethroids . In Ghanethroids , and anyDrosophila melanogaster adapted to high temperatures as compared with those acclimatized to low temperatures [With regard to metabolic enzymes, MFO and GST levels in mosquitoes that were not exposed to pyrethroids increased with increasing temperature from 25 to 32\u00a0\u00b0C. On the other hand, enzyme expression levels decreased at 34\u00a0\u00b0C, suggesting that enzyme expression may be impaired above a certain optimum temperature range . In addieratures . The eleeratures .An. gambiae s.s. mosquitoes exposed to permethrin insecticides were higher than in mosquitoes that were not exposed to insecticides.Comparing the enzyme levels in mosquitoes that were not exposed and those exposed to pyrethroids, levels of MFO, GST, \u03b1-esterase, and \u03b2-esterase were higher in those exposed to pyrethroids than in those that were not exposed, especially at 25 and 30\u00a0\u00b0C. The high resistance of mosquitoes to insecticides might have involved metabolic detoxification due to the elevated enzyme expression in mosquitoes exposed to pyrethroids . These fAn. arabiensis mosquitoes. However, the findings of the current study are contrary to those reported by Leong et al. [A. aegypti mosquitoes. These findings raise the possibility that the levels of metabolic enzymes may differ among mosquito species [The study further revealed that MFO was the least expressed enzyme in mosquitoes. The findings are in agreement with those of Alemayehu et al. , who obsg et al. , who fou species . TherefoAn. gambiae s.l. mosquitoes' susceptibility to insecticides and metabolic enzyme expression. The results suggest that An. gambiae s.l. mosquitoes reared at higher temperatures have increased insecticide resistance than those reared at lower temperatures. In addition, increased rearing temperatures of An. gambiae s.l. mosquitoes were associated with increased expression of metabolic enzymes. Increased metabolic enzyme levels are usually associated with insecticide resistance. This suggests that elevated temperatures projected in a future warmer climate could increase mosquitoes' resistance to insecticides and thus reduce the effectiveness of vector control measures for diseases such as malaria. Therefore, it is essential to research new tools for vector management rather than depending only on chemical insecticides.This study contributes to the knowledge of the effects of temperature variability on Additional file 1: Table S1. Ambient and rearing water conditions for each temperature regime. Table S2. Mortality in An. gambiae s.l. mosquitoes exposed to pyrethroids. Table S3. Median levels of metabolic enzymes in An. gambiae s.l. mosquitoes reared at different temperature regimes. Table S4. Pairwise comparisons of enzyme levels in An. gambiae s.l. mosquitoes reared at different temperature regimes. Table S5. Mann\u2013Whitney U-test between mosquitoes that were not exposed and those exposed to pyrethroids."} +{"text": "H = DSz2 \u2212 \u03b3NV(Sx(BAx + Bx) + Sy(BAy + By) + Sz(BAz + Bz))\u201d. It should read \u201cH = DSz2 \u2212 \u03b3NV(Sx(BAx + Bx) + Sy(BAy + By) + Sz(BAz + Bz))\u201d. On line 7 in the left column on page 180 of the original article, the text originally read \u201cBmin \u2245 4 \u0393 (33 \u03b3NV \u00d7 C)\u22121 (I0 \u00d7 t)\u22121\u201d. It should read \u201cBmin \u2245 4 \u0393 (3\u221a3 \u03b3NV \u00d7 C)\u22121 (I0 \u00d7 t)\u22121/2\u201d. On line 23 in the left column on page 180 of the original article, the text originally read \u201cS = Fl(f1) \u2212 Fl(f2))/(Fl(f1) + Fl(f2)\u201d. It should read \u201cS = (Fl(f1) \u2212 Fl(f2))/(Fl(f1) + Fl(f2))\u201d.The authors regret that there were errors that appear in the Results and discussion section. On line 47 in the right column on page 179, the text originally read \u201cThe authors regret that there was an error in the text in the Author contributions section on line 24 in the right column on page 183 of the original article. The text originally read \u201cASQ\u201d. It should read \u201cSQAS\u201d.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "The hypoxanthinium and xanthinium cations in salts I and II are both in the oxo-N(9)\u2013H tautomeric form. The crystal packing of the two salts is governed predominantly by N\u2014H\u22efO, N\u2014H\u22efBr, C\u2014H\u22efBr and O\u2014H\u22efBr inter\u00adactions described by Two new crystalline salts, namely, hypoxanthinium bromide monohydrate, C 5H5N4O+\u00b7Br\u2212\u00b7H2O (I) and xanthinium bromide monohydrate, C5H5N4O2+\u00b7Br\u2212\u00b7H2O (II), were synthesized and characterized by single-crystal X-ray diffraction technique and Hirshfeld surface analysis. The hypoxanthinium and xanthinium cations in salts I and II are both in the oxo-N(9)\u2013H tautomeric form. The crystal packing of the two salts is governed predominantly by N\u2013H\u22efO, N\u2013H\u22efBr, C\u2013H\u22efBr and O\u2013H\u22efBr inter\u00adactions described by R23(9) and R22(8) synthons. The crystal packing is also consolidated by carbon\u00adyl\u22ef\u03c0 inter\u00adactions between symmetry-related hypoxanthinium (+HX) cations in salt I and xanthinium cations (+XA) in salt II. The combination of all these inter\u00adactions leads to the formation of wave- and staircase-like architectures in salts I and II, respectively. The largest contributions to the overall Hirshfeld surface are from Br\u22efH/H\u22efBr contacts (22.3% in I and 25.4% in II) .Two new crystalline salts, namely, hypoxanthinium bromide monohydrate, C Similarly, xanthinium nitrate monohydrate, xanthinium hydrogensulfate monohydrate crystallizes in the monoclinic space group P21/c with one hypoxanthinium cation (+HX), one bromide anion (\u2212Br) and one water mol\u00adecule in the asymmetric unit, as shown in Fig.\u00a01+HX cation exists in the oxo-N(9)\u2013H tautomeric form with the N7 atom of the purine ring protonated, as can be seen from the N\u2014C bond distance [N7\u2014C8 = 1.3219\u2005(17)\u2005\u00c5 vs N9\u2014C8 = 1.3419\u2005(18)\u2005\u00c5] and C\u2014N\u2014C bond angles [C5\u2014N7\u2014C8 = 107.98\u2005(11)\u00b0 and C4\u2014N9\u2014C8 = 108.32\u2005(10)\u00b0]. Those values are similar to those in the crystal structure of hypoxanthinium chloride monohydrate shorter than N9\u2013C8 one [1.344\u2005(5)\u2005\u00c5]. The C\u2014N\u2014C bond angles are C5\u2014N7\u2014C8 = 108.2\u2005(3)\u00b0 and C4\u2014N9\u2014C8 = 107.7\u2005(3)\u00b0 and, therefore, the cation can also be described as the oxo-N(9)\u2013H tautomer. These values are similar to those in xanthinium perchlorate dihydrate . The dimer is flanked on both sides by a water mol\u00adecule (O1W), forming a pair of O1W\u2014H2W\u22efO2iv and O1W\u2014H1W\u22efO6ii hydrogen bonds with an HDADA array along the ac plane. Neighbouring tetra\u00admeric units are then connected through two sets of W and O1W\u2014H1W\u22efO6ii hydrogen bonds and an L) motif formed by a pair of O1W\u2014H1W\u22efO6ii inter\u00adactions. The tetra\u00admeric units combine into a supra\u00admolecular ribbon extended along the ac plane of the +XA unit) with C=O\u22efCg2vi and C=O\u22efCg2vii distances of 3.366\u2005(3) and 3.477\u2005(3)\u2005\u00c5, respectively, and angles of 108.2\u2005(2) and 118.7\u2005(2)\u00b0 [symmetry codes: (vi) 1\u00a0+\u00a0x, y, z; (vii) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z; Fig.\u00a08In the crystal structure of salt s Table\u00a02 with an ne Fig.\u00a05. Neighbo4.et al., 2007Crystal Explorer 17.5 inter\u00adactions or shorter (red) than the sum of the van der Waals radii of the two inter\u00adacting atoms. The Hirshfeld surfaces of salts I and II are shown in Fig.\u00a09a and 10aI and II and the respective neighbouring moieties are shown in Fig.\u00a09b and 10bI and II are shown in Figs. 11I 19.7%, II 23.4%), N\u22efH/H\u22efN C\u22efH/H\u22efC , H\u22efH/H\u22efH and C\u22efC/C\u22efC (Table\u00a05), indicating that the most abundant contact is Br\u22efH/H\u22efBr with 22.3% in I and 25.4% in II, respectively.Hirshfeld surface analyses and their associated two-dimensional fingerprint plots and xanthinium bromide monohydrate (II) using the following qu\u00adanti\u00adties: 0.0340\u2005mg (0.25mmol) of hypoxanthine for I and 0.0380\u2005mg (0.25\u2005mmol) of xanthine for II.A general method was used for the preparation and crystallization of the hypoxanthinium bromide monohydrate (The indicated amount of the base was dissolved in 20\u2005mL of distilled water and 2\u2005mL of hydro\u00adbromic acid (5% in water) were added. The reaction mixture was heated to 358\u2005K for 30\u2005min using a water bath. The resulting solution was allowed to slowly evaporate at room temperature. After a few days, colourless plate-like crystals were obtained.8.I and II are summarized in Table\u00a04Uiso(H) = 1.2Ueq (C). The H atoms of the water mol\u00adecule were located in a difference-Fourier map and refined with the O\u2014H distance restrained to 0.85\u20130.86\u2005\u00c5 and with Uiso(H) = 1.5 Ueq(O). The hydrogen atoms bound to the nitro\u00adgen atoms in salts I and II were located in difference-Fourier maps and either refined freely (in I) or with the distance restraint N\u2014H = 0.82\u2005\u00c5 and with Uiso(H) = 1.2Ueq(N) (in II).Crystal data, data collection and structure refinement details for salts 10.1107/S2056989022005278/jq2017sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989022005278/jq2017Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022005278/jq2017Isup4.cmlSupporting information file. DOI: 10.1107/S2056989022005278/jq2017IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989022005278/jq2017IIsup5.cmlSupporting information file. DOI: 2170923, 2170922CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds link mol\u00adecules in the crystal into layers parallel to (011). The crystal packing is consolidated through C\u2014Br\u22ef\u03c0 and C\u2014F\u22ef\u03c0 inter\u00adactions, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions. 14H8Br2FN3O2, the 4-fluoro\u00adphenyl ring and the nitro-substituted phenyl ring form a dihedral angle of 64.37\u2005(10)\u00b0. Mol\u00adecules in the crystal are connected by C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds into layers parallel to (011). The crystal packing is consolidated by C\u2014Br\u22ef\u03c0 and C\u2014F\u22ef\u03c0 inter\u00adactions, as well as by \u03c0\u2013\u03c0 stacking inter\u00adactions. According to a Hirshfeld surface analysis of the crystal structure, the most significant contributions to the crystal packing are from O\u22efH/H\u22efO (15.0%), H\u22efH (14.3%), Br\u22efH/H\u22efBr (14.2%), C\u22efH/H\u22efC (10.1%), F\u22efH/H\u22efF (7.9%), Br\u22efBr (7.2%) and Br\u22efC/C\u22efBr (5.8%) contacts.In the title compound, C Depending on the attached substituents, azo compounds have attracted attention because of their high synthetic potential for organic and inorganic chemistry and numerous useful properties. For example, azo dyes find applications in the design of functional materials attributed to smart hydrogen bonding, as self-assembled layers, photo-triggered structural switching, liquid crystals, ionophors, indicators, semiconductors, spectrophotometric reagents for determination of metal ions, catalysts, photoluminescent materials, optical recording media, spin-coating films and anti\u00admicrobial agents -1--2-(4-fluoro\u00adphen\u00adyl)diazene, the mol\u00adecular and crystal structure of which along with a Hirshfeld surface analysis are reported here.In the above context, we have attached F, Br and NO2.trans-configured with regard to the N=N double bond and are practically coplanar as revealed by the C2\u2014N2=N3\u2014C9 torsion angle of \u2212178.63\u2005(16)\u00b0. All of the bond lengths and angles in the title compound are similar to those for the related azo compounds reported in the Database survey section.The mol\u00adecular conformation of the title compound is not planar, as seen in Fig.\u00a013.Cg1 = 3.6016\u2005(9)\u2005\u00c5, C1\u2014Br1\u22efCg1 = 104.24\u2005(7)\u00b0] and C\u2014F\u22ef\u03c0 inter\u00adactions, and weak \u03c0\u2013\u03c0 stacking , where Cg1 and Cg2 are the centroids of the C3\u2013C8 and C9\u2013C14 rings, respectively, Table\u00a01\u20134 \u25b8 \u25b8. 4.Crystal Explorer 17.5 to 1.1463 (blue) a.u. -1--2-phenyl\u00addiazene moiety resulted in 27 hits. Eight compounds are closely related to the title compound, viz. those with CSD refcodes GUPHIL (I) , mol\u00adecules are linked into inversion dimers via short halogen\u22efhalogen contacts compared to the van der Waals radius sum of 3.50\u2005\u00c5 for two chlorine atoms. No other directional contacts could be identified, and the shortest aromatic ring centroid separation is greater than 5.25\u2005\u00c5. In the crystals of (II) and (III), mol\u00adecules are linked through weak X\u22efCl contacts [X = Cl for (II) and Br for (III)], C\u2014H\u22efCl and C\u2014Cl\u22ef\u03c0 inter\u00adactions into sheets lying parallel to (001). In the crystal of (IV), mol\u00adecules are stacked in columns parallel to [100] via weak C\u2014H\u22efCl hydrogen bonds and face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions. The crystal packing is further consolidated by short Cl\u22efCl contacts. In (V), mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds into zigzag chains running parallel to [001]. The crystal packing also features C\u2014Cl\u22ef\u03c0, C\u2014F\u22ef\u03c0 and N\u2014O\u22ef\u03c0 inter\u00adactions. In (VI), C\u2014H\u22efN and short Cl\u22efCl contacts are observed, and in (VII), C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and short Cl\u22efO contacts occur. In the crystal of (VIII), mol\u00adecules are linked into chains running parallel to [001] by C\u2014H\u22efO hydrogen bonds. The crystal packing is consolidated by C\u2014F\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions, and short Br\u22efO [2.9828\u2005(13)\u2005\u00c5] contacts are also observed.In the crystal of (6.ababE)-1-(4-fluoro\u00adphen\u00adyl)-2-(4-nitro\u00adbenzyl\u00adidene)hydrazine (1\u2005mmol), tetra\u00admethyl\u00adethylenedi\u00adamine , CuCl and CBr4 (4.5\u2005mmol). After 1\u20133\u2005h , the reaction mixture was poured into a 0.01\u2005M HCl solution , and extracted with di\u00adchloro\u00admethane (3 \u00d7 20\u2005ml). The combined organic phase was washed with water (3 \u00d7 50\u2005ml), brine (30\u2005ml), dried over anhydrous Na2SO4 and concentrated in vacuo using a rotary evaporator. The residue was purified by column chromatography on silica gel using appropriate mixtures of hexane and di\u00adchloro\u00admethane (v/v 3/1\u20131/1). Light-orange solid (yield 52%); m.p. 377\u2005K. Analysis calculated for C14H8Br2FN3O2 (M = 429.04): C 39.19, H 1.88, N 9.79; found: C 39.17, H 1.85, N 9.76%. 1H NMR \u03b4 7.36\u20137.14 . 13C NMR \u03b4 164.35, 153.13, 152.46, 133.69, 133.24, 131.74, 127.98, 127.89, 127.75, 127.42, 119.07, 89.02. ESI\u2013MS: m/z: 430.06 [M\u00a0+\u00a0H]+. Crystals suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution.The title compound was synthesized according to a reported method = 1.2Ueq(C). The maximum electron density in the final difference map is located 0.75\u2005\u00c5 from atom Br1, while the minimum electron density is located 0.72\u2005\u00c5 from Br2.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989022004388/wm5642sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022004388/wm5642Isup2.hklStructure factors: contains datablock(s) I. DOI: 2168678CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Colorless, block-shaped crystals were grown in a potassium hydro\u00adflux, i.e. under ultra-alkaline conditions, within 10\u2005h. K2SeO3 crystallizes isostructural with Na2SO3 and K2TeO3 in the trigonal space group Pa = 6.1063\u2005(4)\u2005\u00c5 and c = 6.9242\u2005(4)\u2005\u00c5 at 100\u2005(1)\u2005K. In the trigonal\u2013pyramidal, C3v-symmetric [SeO3]2\u2013 anion, the bond length is 1.687\u2005(1)\u2005\u00c5, and the bond angle is 103.0\u2005(1)\u00b0. Two of the three unique potassium cations exhibit a coordination number of six, and the third a coordination number of nine.Crystal structure data for potassium orthoselenate(IV), K K2Se2O5 are a long-known but poorly studied class of compounds. After the discovery of the first salts of selenic acid by Berzelius, comprehensive studies on these salts were not carried out until the beginning of the 1930s, when Janitzki reported the syntheses of sodium and potassium salts of selenic acid 2] , K2.2SeO3, one selenium atom , three potassium atoms and one oxygen atom . The unit cell of K2SeO3 is depicted in Fig.\u00a01C3v-symmetric [SeO3]2\u2013 anion can be attributed to the electron lone pair of the selenium(IV) atom. This oxidation state is supported by the bond-valence sum calculation = \u2211exp\u2005[(RSeO \u2013 dSeO)/b)] = 3 \u00b7 exp\u2005[(1.811\u2005\u00c5 \u2013 1.687\u2005(1)\u2005\u00c5) / 0.37\u2005\u00c5)] = 4.2 valence units. The potassium cations K1 and K2 are octa\u00adhedrally coordinated by oxygen atoms with K\u2014O distances of 2.631\u2005(1) and 2.771\u2005(1)\u2005\u00c5, respectively. K3 has nine oxygen neighbors at distances of 2.792\u2005(1), 3.020\u2005(1)\u2005\u00c5, and 3.474\u2005(1)\u2005\u00c5 groups is responsible for the symmetry reduction to PK4.2SeO3, was synthesized in a potassium hydroxide hydro\u00adflux with a molar water-base ratio of 1.7. The reaction was carried out in a PTFE-lined 50\u2005mL Berghof-type DAB-2 stainless steel autoclave to prevent evaporation of water. The starting material SeO2 was dissolved in 3\u2005ml of water before adding 6.3\u2005g of KOH . After closing the autoclave, the reaction mixture was heated to 473\u2005K at a rate of 2\u2005K\u2005min\u22121 and, after 8\u2005h, cooled to room temperature at a rate of \u22121\u2005K\u2005min\u22121. The solid reaction product was washed twice with 2\u2005ml of methanol on a Schlenk frit under inert conditions to remove adherent hydro\u00adflux. The colorless, block-shaped crystals of K2SeO3 , KO3 Fig.\u00a04 dissolveO3 Fig.\u00a04. Due to JANA2006 was used . The composition of selected single crystals was determined by semi-qu\u00adanti\u00adtative energy dispersive X-ray analysis (Ua = 15\u2005kV) using a Silicon Drift Detector X\u2013MaxN (Oxford Instruments). The data were processed applying the AZtec software package global, I. DOI: 10.1107/S2056989022005175/wm5645Isup2.hklStructure factors: contains datablock(s) I. DOI: 2172487CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Correction: BMC Neuroscience (2022) 23:68 10.1186/s12868-022-00754-4fMRI data acquisition section.Following the publication of the original article , an errobold typeface.The updated text is given below, and the changes have been highlighted in fMRI data acquisition8\u00b0, 256\u2009\u00d7\u2009256 matrix, voxel size 0.7\u2009\u00d7\u20090.7\u2009\u00d7\u20090.7\u00a0mm3. For the resting state scans before and after stress induction (RS1 and RS2), 355 volumes of T2*-weighted echo-planar images were acquired for each session with the following parameters: 34 axial slices covering the whole brain, repetition time\u2009=\u20092000\u00a0ms, echo time\u2009=\u200930\u00a0ms, flip angle\u2009=\u200990\u00b0, 9696 matrix, field of view\u2009=\u2009240,240\u00a0mm2, voxel size\u2009=\u20092.5\u2009\u00d7\u20092.5\u2009\u00d7\u20093\u00a0mm3.A Philips 3T scanner was used for fMRI data acquisition. Structural T1-weighted images for spatial normalization were measured using a turbo field echo sequence with the following parameters: 274 sagittal slices covering the whole brain, flip angle\u2009=\u2009Moreover, the authors identified an error in Table 1. The correct Table Finally, the authors identified an error in Fig. 4. The correct Fig.\u00a0The original article has been"} +{"text": "E responded in opposite manner 5\u00a0min post\u2010exercise compared to E\u2009+\u2009C (p\u00a0=\u00a00.013) and C (p\u00a0=\u00a00.032) in routine respiration, and to E\u2009+\u2009C in free routine activity (p\u00a0=\u00a00.013). E\u2009+\u2009C\u2009>\u2009C\u2009>\u2009E was observed for increased lactate levels and decreased isometric force that correlated with routine respiration . Lactate and pH change did not affect bioenergetics of PBMCs. Acute resistance exercise affected cellular respiration of PBMCs, with training volume and the amount of metabolic stress appear influential. Results suggest that acute inflammation response does not contribute to changes seen in cellular respiration, but the level of peripheral muscle fatigue and metabolic stress could be explaining factors.Little is known how acute exercise\u2010induced inflammation and metabolic stress affect immune cell bioenergetics and the portion of its components. Therefore, we investigated acute effects of eccentric\u2010only (E), concentric\u2010only (C) and combined eccentric\u2010concentric resistance exercise (E\u2009+\u2009C) bouts on cellular respiration of peripheral blood mononuclear cells (PBMCs). Twelve strength\u2010trained young men performed bench press resistance exercises in randomized order. Venous blood samples were drawn at pre\u2010, 5\u00a0min post\u2010 and 24\u2009h post\u2010exercise. Several PBMC respiration states were measured using high\u2010resolution respirometry. Levels of leukocytes, interleukin 6 (IL\u20106), C\u2010reactive protein (CRP), creatine kinase (CK), blood lactate and maximum voluntary isometric force were measured from the same time points. Effects of blood lactate and pH change on bioenergetics of PBMCs were investigated ex vivo. PBMC routine respiration ( Resistance exercise with different workloads have distinct effect on cellular respiration of PBMCs. Training volume and amount of metabolic stress are influential. However, an exercise model comparing responses to eccentric and concentric exercise could reveal differences in bioenergetics of PBMCs accompanying the expected differences in metabolic cost and inflammation response. Consequently, our study investigated the acute effects of three resistance exercise bouts, eccentric\u2010only (E), concentric\u2010only (C) and combined eccentric and concentric contractions (E\u2009+\u2009C), on cellular respiration of PBMCs. It was hypothesized that metabolic cost and inflammation response to C versus E are different and would lead to different respiration responses of PBMCs. Combined eccentric and concentric contractions would reveal whether high metabolic cost and high inflammation response has an additive effect on cellular respiration of PBMCs. Finally, we studied ex vivo if lactate concentration and change in pH specifically affects cellular respiration of PBMCs.22.1Twelve healthy male subjects were recruited to the study through local advertisement . Inclusion criteria were experience in resistance training (>100\u2009kg bench press or \u22651.25 times bodyweight). We hypothesized that inflammation response in untrained test subjects might vary largely, therefore only trained test subjects were chosen to obtain more homogenous responses. In addition, inclusion criteria included no use of anabolic agents such as steroids or medication known to affect the inflammatory status.n with calculated effect sizes for study design by using G*Power program with 80% power.We estimated the appropriate n\u00a0=\u00a011.All subjects gave their written informed consent prior to participation. All participants filled a PAR\u2010Q+ questionnaire . All exercise protocols were completed with this isokinetic bench press device. The machine operated with a speed of 0.1\u00a0m/s in all three exercises, independent of the force produced against it. Ground reaction forces were recorded with a 2\u2010D strain gauge force plate positioned under the bench. During the concentric and eccentric loadings, the bar's movement paused for 500\u2009ms at the top and bottom position, to aid pacing the contractions. In E, subjects were asked to start to maximally contract their muscles, once the bar stopped at the top position and continue pushing as hard as possible until the bar stopped at the bottom position . In C, subjects were asked to maximally contract their muscle once the bar stopped at the bottom position and continue pushing until the bar stopped at the up position. During the unloaded traveling phase of the machine, subjects were instructed to rest their arms while grasping the bar to avoid activation of the targeted muscles. During E\u2009+\u2009C, only 1\u00a0ms \u20182.4Maximal isometric force was recorded at 90\u00b0 of elbow flexion, measured with a custom\u2010made isometric bench press were drawn from the antecubital vein from each subject while seated pre\u2010, 5\u00a0min post\u2010 and 24\u2009h post\u2010exercise to Vacutainer VACUETTE K3EDTA (Greiner Bio\u2010One) tubes containing anticoagulant (EDTA). Whole blood count and the number of lymphocytes and monocytes were assessed by Sysmex XP\u2010300 (Sysmex Corporation) to obtain the number of leukocytes and platelets. 18\u2009ml of blood was used for isolation of PBMCs and 12\u00a0ml for serum isolation.g). Serum was stored in \u221220\u00b0C for later analysis, which was performed using IMMULITE 2000 XPi Immunoassay System according to manufacturers\u00b4 instructions. Lactate was measured from fingertip blood samples, which were collected into capillary tubes (20\u2009\u03bcl) at all three time points. Samples were then placed in a 1\u2009ml hemolyzing solution and analyzed automatically with Biosen C\u2010line lactate analyzer .For the detection of IL\u20106, CK and CRP, serum was isolated from blood samples by centrifugation . Standing time did not have a statistical effect on cellular respiration.g . The layer of PBMCs and platelets was collected and washed twice with RPMI\u20101640 in order to reduce PLT/PBMC ratio below 7 with 15\u00a0ml of Ficoll\u2010Paque TM PLUS . Substrate\u2010uncoupler\u2010inhibitor titration 3 (SUIT3) protocol designed for intact cells was chosen for measurements from five resistance trained men and isolated PBMCs and measured the cellular respiration by using the same methods as described above.To test whether lactate had an impact on the cellular respiration of PBMCs, 20\u2009mmol of sodium L\u2010lactate (Sigma Aldrich) was added to the HRR measurement chamber after measurements of routine respiration of PBMCs and SUIT3 protocol was completed \u2010lactic acid (Sigma Aldrich) to the HRR measurement chamber after routine respiration of PBMCs and SUIT3 protocol was completed. According to studies, a vigorous exercise can drop the pH of blood near 7.10 and could be under 7.0 in skeletal muscle .Statistical analyses were carried out using the Statistical Package for Social Sciences . Values are reported as mean\u2009\u00b1\u2009standard deviation (SD), unless otherwise stated. All values were verified for outliers, which were excluded from the analysis (>3\u00d7 interquartile range [IQR]). Significance level was set at 0.05 in all analyses. ANOVA with repeated measures was used to examine whether there were differences in the study variables between the groups over time . Mauchly's test was used to test the assumption of sphericity and Greenhouse\u2013Geisser corrections were applied if the assumption was violated. To study the effect of time separately within resistance exercises and group\u2009\u00d7\u2009time interaction between adjacent time points, within\u2010subject contrasts were used as post hoc comparison. Hedges' 33.1Cellular respiration of PBMCs was analyzed through several respiration states Table\u00a0, which i1)\u00a0=\u00a08.2, p\u00a0=\u00a00.017, g\u2009=\u2009\u22120.83), free routine activity (F(1)\u00a0=\u00a06.8, p\u00a0=\u00a00.025, g\u2009=\u2009\u22120.73) and ET\u2010 capacity (F(1)\u00a0=\u00a05.7, p\u00a0=\u00a00.038, g\u2009=\u2009\u22120.69) decreased significantly immediately after E\u2009+\u2009C \u00a0=\u00a010.8, p\u00a0=\u00a00.008, g\u2009=\u00a00.95) \u00a0=\u00a06.5, p\u00a0=\u00a00.032, g\u2009=\u00a00.73) and E\u2009+\u2009C (F(1)\u00a0=\u00a09.6, p\u00a0=\u00a00.013, g\u2009=\u00a00.94) in routine respiration \u00a0=\u00a09.5, p\u00a0=\u00a00.013, g\u2009=\u00a00.87) in respiration states immediately after resistance exercise (from pre\u2010exercise to 5\u00a0min post\u2010exercise response) were compared between resistance exercises Figure\u00a0. Post hon Figure\u00a0. Free ro) Figure\u00a0.1)\u00a0=\u00a07.4, p\u00a0=\u00a00.026, g\u2009=\u00a01.17) .Main effects of group and time, and their interaction did not show statistical significances on CRP, CK or IL\u20106, hence there were no major changes between time points or between resistance exercises. Mean values and SDs of the data are represented in supplementary data \u00a0=\u00a089.3, p\u2009<\u20090.001, g\u2009=\u2009\u22122.74; E: \u221222\u2009\u00b1\u20096%, F(1)\u00a0=\u00a0131.2, p\u2009<\u20090.001, g\u2009=\u2009\u22123.32; E\u2009+\u2009C: \u221241\u2009\u00b1\u200910%, F(1)\u00a0=\u00a0160.6, p\u2009<\u20090.001, g\u2009=\u2009\u22123.84) and this was reversed at 24\u2009h post\u2010exercise \u00a0=\u00a046.1, p\u2009<\u20090.001, g\u2009=\u2009\u22122.14) and C (F(1)\u00a0=\u00a06.4, p\u00a0=\u00a00.032, g\u2009=\u2009\u22120.79) showed a statistically greater decrease than E \u00a0=\u00a013.8, p\u00a0=\u00a00.005, g\u2009=\u00a01.19) in isometric force (from pre\u2010 to 5\u00a0min post\u2010exercise response) were compared between resistance exercises Figure\u00a0. E\u2009+\u2009C Figure\u00a0.3.2.31)\u00a0=\u00a0103.1, p\u2009<\u20090.001, g\u2009=\u00a03.08; E: F(1)\u00a0=\u00a064.0, p\u2009<\u20090.001, g\u2009=\u00a02.32; E\u2009+\u2009C: F(1)\u00a0=\u00a0171.6, p\u2009<\u20090.001, g\u2009=\u00a03.80) Figure\u00a0 and this1)\u00a0=\u00a014.8, p\u00a0=\u00a00.004, g\u2009=\u00a01.27) and C (F(1)\u00a0=\u00a05.3, p\u00a0=\u00a00.047, g\u2009=\u00a00.70) showed a statistically greater increase than E. Also, the increase from E\u2009+\u2009C was greater than the increase from C (F(1)\u00a0=\u00a05.2, p\u00a0=\u00a00.048, g\u2009=\u00a00.69) in lactate levels (from pre\u2010 to 5\u00a0min post\u2010exercise response) were compared between resistance exercises Figure\u00a0. E\u2009+\u2009C Figure\u00a0.3.3Pearson correlation coefficients for 5\u00a0min post\u2010exercise response in routine respiration, free routine activity and ET\u2010capacity in relation to 5\u00a0min post\u2010exercise response in lactate, isometric force and inflammation and muscle injury markers are presented in Table\u00a03.43)\u00a0=\u00a0\u22123.583, p\u00a0=\u00a00.035) Figure\u00a0.3.5WBCs, as well as lymphocytes and monocytes, which are the main components of PBMCs, were counted at each time point and analyzed between timepoints and resistance exercises. The effect of plasma volume change was examined (according to Dill & Costill 1974] equation) \u00a0=\u00a037.4, p\u2009<\u20090.001, g\u2009=\u00a01.70; E: F(1)\u00a0=\u00a09.9, p\u00a0=\u00a00.010, g\u2009=\u00a00.91; E\u2009+\u2009C: F(1)\u00a0=\u00a018.7, p\u00a0=\u00a00.001, g\u2009=\u00a01.20) (Table\u00a01)\u00a0=\u00a05.7, p\u00a0=\u00a00.039, g\u2009=\u2009\u22120.72) and E\u2009+\u2009C (F(1)\u00a0=\u00a09.0, p\u00a0=\u00a00.012, g\u2009=\u2009\u22120.84). In C, the difference between pre\u2010 and 24\u2009h post\u2010exercise WBC counts were also significantly different (F(1)\u00a0=\u00a014.0, p\u00a0=\u00a00.003, g\u2009=\u00a01.04) Table\u00a0. The dif1)\u00a0=\u00a018.6, p\u00a0=\u00a00.001, g\u2009=\u00a01.16; E: F(1)\u00a0=\u00a08.8, p\u00a0=\u00a00.014, g\u2009=\u00a00.83; E\u2009+\u2009C: F(1)\u00a0=\u00a020.6, p\u00a0=\u00a00.001, g\u2009=\u00a01.22) (Table\u00a01)\u00a0=\u00a011.7, p\u00a0=\u00a00.006, g\u2009=\u2009\u22120.92). The difference between pre\u2010 and 24\u2009h post\u2010exercise lymphocyte count was significantly different in C (F(1)\u00a0=\u00a011.1, p\u00a0=\u00a00.007, g\u2009=\u00a00.89). The number of monocytes was elevated after C (F(1)\u00a0=\u00a06.5, p\u00a0=\u00a00.027, g\u2009=\u00a00.68) and E (F(1)\u00a0=\u00a05.3, p\u00a0=\u00a00.045, g\u2009=\u00a00.64) and pre\u2010 and 24\u2009h post\u2010exercise count reached statistically significant difference in E (F(1)\u00a0=\u00a07.7, p\u00a0=\u00a00.019, g\u2009=\u00a00.77).Each resistance exercise elevated the number of lymphocytes (C: F(2) Table\u00a0. The dif1)\u00a0=\u00a011.7, p\u00a0=\u00a00.007, g\u00a0=\u00a00.66) and E (F(1)\u00a0=\u00a05.0, p\u00a0=\u00a00.048, g\u00a0=\u00a00.55). The number of lymphocytes was significantly higher after E\u2009+\u2009C when compared to C (F(1)\u00a0=\u00a010.1, p\u00a0=\u00a00.010, g\u00a0=\u00a00.66). There were no significant differences in the response between exercises among monocytes.Change (\u2206) in WBC, lymphocyte and monocyte counts (5\u00a0min post\u2010 exercise response) were compared between resistance exercises contractions caused an acute decrease in cellular respiration of PBMCs as assessed by routine respiration, free routine capacity and ET\u2010capacity Figure\u00a0. ResultsChanges seen in routine control ratio were comparable with respiration results normalized to cell concentration Figure\u00a0. E\u2009+\u2009C c4.1PBMCs have a central role in muscle repair and exercise\u2010related anti\u2010inflammatory response by producing anti\u2010inflammatory cytokines were somewhat lower than a previous study (10\u201312\u2009mmoL/L) correlated negatively with the change in routine respiration Table\u00a0, indicat4.3Peripheral muscle fatigue is a result of several mechanisms including metabolic acidosis and reduction of ATP axis eventually leading to elevated cortisol levels exercise\u2010induced inflammation nor (2) anaerobic metabolites were contributing factors to the acute respiration responses.Experimental design: M.L., S.W., E.I.L., M.K. and I.K. Investigation and data collection: E.I.L, M.K., I.K., E.L. and A.K. Writing and manuscript preparation: E.I.L, M.K., A.K., H.K., S.W. and M.L. Review and editing of the manuscript: S.W., M.L., E.I.L. and H.K. Funding acquisition: H.K. All authors have read and agreed the final version of the manuscript.No conflicts of interest are declared by the authors."} +{"text": "The title compound arose from an unexpected alcoholysis of a prodrug by the methanol solvent. 14H13ClN2O2, was obtained during an attempt to grow single crystals of 4-acetyl\u00adphenyl 2-[(3-chloro-2-methyl\u00adphen\u00adyl)amino]\u00adnicotinate in methanol, and was probably generated by alcoholysis. Two intra\u00admolecular hydrogen bonds are formed, one between the N\u2014H group and the carbonyl O atom of the ester and the other between the ortho sp2CH group of the benzene ring and the pyridine N atom. Aromatic \u03c0\u2013\u03c0 stacking [shortest centroid\u2013centroid separation = 3.598\u2005(2)\u2005\u00c5] is observed in the extended structure.The title compound, C I) was first synthesized when preparing esters of anthranilic acid as possible analgesic and anti-inflammatory agents \u00b0 between the C1\u2013C6 benzene and N2/C8\u2013C12 pyridine rings \u2005\u00c5]: both of these close S(6) rings. The cohesion of the crystal structure is ensured by aromatic \u03c0\u2013\u03c0 stacking between the benzene and pyridine rings [shortest centroid\u2013centroid separation = 3.598\u2005(2)\u2005\u00c5] and hydro\u00adphobic inter\u00adactions amino]\u00adnicotin\u00adate, synthesized by a condensation reaction between clonixin and paracetamol global, I. DOI: 10.1107/S2414314621005393/hb4383Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314621005393/hb4383Isup3.cmlSupporting information file. DOI: 2085247CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of the title compound is consolidated by N\u2014H\u22efCl and C\u2014H\u22efCl hydrogen bonding inter\u00adactions. 2H8N2)3][ZnCl4], exists as discrete ions. The [Zn(C2H8N2)3]2+ cation exhibits a distorted octa\u00adhedral shape. In the [ZnCl4]2\u2212 anion, the ZnII atom is in an almost regular tetra\u00adhedral environment. The crystal packing is consolidated by N\u2014H\u22efCl and C\u2014H\u22efCl hydrogen bonds.The title complex, [Zn(C X hydrogen bonds. Metal complexes containing an ethyl\u00adenedi\u00adamine (\u2013NCH2CH2N) backbone have attracted significant inter\u00adest as potential anti\u00adcancer agents because of their rich redox chemistry and relative ease of manipulation is a common chelating ligand that is widely used in transition-metal complexes. It cannot only chelate metal cations by two nitro\u00adgen atoms, but also offers hydrogen atoms to form N\u2014H\u22efMetals such as zinc act as a key structural component in many proteins and enzymes, including transcription factors, cellular signalling proteins and DNA repair enzymes 3][ZnCl4], which comprises an ZnCl42\u2212 anion and a [Zn(en)3]2+complex cation. The ZnII atom of the tetra\u00adchlorido\u00adzincate(II) anion is in an almost regular Cl4 tetra\u00adhedral environment, with Zn\u2014Cl bond lengths in the range 2.255\u2005(1)-2.272\u2005(9)\u2005\u00c5. The zinc cation displays a distorted octa\u00adhedral coordination geometry defined by six N atoms from three ethyl\u00adenedi\u00adamine ligands, with Zn\u2014N distances in the range of 2.173\u2005(3)\u20132.219\u2005(3)\u2005\u00c5. The N\u2014Zn\u2014N angles of the en ligands are about 80\u00b0. They are noticeably smaller than the ideal octa\u00adhedral angle of 90\u00b0. The five-membered chelate rings are non-planar, with N\u2014C\u2014C\u2014N torsion angles of \u221257.5\u2005(4), \u221255.4\u2005(4) and \u221255.9\u2005(5)\u00b0. All of the three en ligands assume a synclinal conformation about the C\u2014C bond.Fig.\u00a01via inter\u00admolecular hydrogen bonds. The N\u2014H\u22efCl hydrogen-bonding inter\u00adactions between the N atoms of the ethyl\u00adenedi\u00adamine ligands and Cl atoms of the tetra\u00adchlorido\u00adzincate anion connect the mol\u00adecules, together with the weak C\u2014H\u22efCl intra\u00admolecular inter\u00adactions, generating a three-dimensional network mixture. To this solution, ethyl\u00adenedi\u00adamine in 25\u2005ml of an HCl/EtOH (2:3 v/v) mixture was added dropwise. The mixture was stirred and heated to 338\u2005K for 2\u2005h and allowed to stand at room temperature until colourless crystals separated (3\u20134 weeks). Crystals suitable for single-crystal XRD were collected after recrystallization using acidified water.Zinc chloride was dissolved in 25\u2005ml of EtOH/HCrystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314620006185/bt4091sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2414314620006185/bt4091Isup2.hklStructure factors: contains datablock(s) I. DOI: 2001547CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In Table 1, the fraction of p\u2010c\u2010Jun+neurons in NeuroTrace+ neurons for AR group (Estimate and 95%CI) is shown as \u201c10.3% (10.1\u201310.5%).\u201d This should be \u201c13.0% (12.8\u201313.3%).\u201d The corrected Table 1 is shown below:In Table S3, the fraction of p\u2010c\u2010Jun+neuronal population for AR group (Estimate and 95%CI) is shown as \u201c10.3% (10.1\u201310.5%).\u201d This should be \u201c13.0% (12.8\u201313.3%).\u201d Corrected version of Table S3 is corrected online.In the article by Ogino et al.,"} +{"text": "The structure of hexa\u00adaqua\u00adnickel(II) bis\u00ad(3-carb\u00adoxy-4-hy\u00addroxy\u00adbenzene\u00adsulfonate) dihydrate consists of alternating layers of inorganic cations and organic anions linked by O\u2014H\u22efO hydrogen bonds that also include non-coordinated water mol\u00adecules of crystallization. The structure of hexa\u00adaqua\u00adcobalt(II) bis\u00ad(3-carb\u00adoxy\u00adbenzene\u00adsulfonate) dihydrate is also built of alternating layers of complex cations and organic anions without direct coordination to the metal by the protonated carboxyl\u00adate or unprotonated sulfonate groups. A robust O\u2014H\u22efO hydrogen-bonding network involving primarily the coordinated and non-coordinated water mol\u00adecules and sulfonate groups directs the packing. 2O)6][C6H3(CO2H)(OH)SO3]2\u00b72H2O, (I), crystallizes in the triclinic space group P2O)6][C6H4(CO2H)SO3]2\u00b72H2O, (II), also crystallizes in triclinic PHexa\u00adaqua\u00adnickel(II) bis\u00ad(3-carb\u00adoxy-4-hy\u00addroxy\u00adbenzene\u00adsulfonate) dihydrate, [Ni(H As a result of the symmetry, the nickel ion has a very regular octa\u00adhedral coordination of six water mol\u00adecules \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a02, \u2212z], and two in which the water hydrogen atoms H41 and H42 are donors to sulfonate oxygen atoms O1 and O2, respectively.The product of the reaction of nickel nitrate hexa\u00adhydrate and 5-sulfosalicylic acid (3-carb\u00adoxy-4-hy\u00addroxy\u00adbenzene\u00adsulfonic acid) is [Ni6](C6H4(CO2H)SO3)2\u00b72H2O, (II)Pet al., 1993vs C7\u2014O5 = 1.213\u2005(2)\u2005\u00c5] and rotated slightly out of the plane of the ring [torsion angle C2\u2014C3\u2014C7\u2014O4 = 4.6\u2005(2)\u00b0]. The sulfon\u00adate group is rotated about 19\u00b0 from its position in (I)vs \u221225.29\u2005(12)\u00b0 in (I)]. Presumably this difference is driven by the hydrogen-bonding patterns. The non-coordinated water mol\u00adecule has a different hydrogen-bonding environment , respectively. These inter\u00adactions are somewhat longer and less linear than those seen in (I)The reaction of cobalt nitrate hexa\u00adhydrate and sodium 3-sulfobenzoate (3-carb\u00adoxy\u00adbenzene\u00adsulfonate) produced crystals that have been identified as (C6H3(CO2H)(OH)SO3)2\u00b74H2O structure has also been reported for cobalt 6](C6H3(CO2H)(OH)SO3)2\u00b72H2O have been reported for manganese salt of 3-carb\u00adoxy-4-hy\u00addroxy\u00adbenzene\u00adsulfonate has been reported previously ns Fig.\u00a04. Two watns Fig.\u00a04. The nonns Fig.\u00a04, and as ns Fig.\u00a04. Other O4.et al., 2016et al., 2005aet al., 2004et al., 2003det al., 2005et al., 2003bet al., 2003aet al., 2003eet al., 2003cet al., 2011et al., 2019A search of the Cambridge Structural Database 2.6H2O (Aldrich) in 50\u2005ml of water. The resulting clear green solution was stirred for about 30 minutes and transferred to a porcelain evaporating dish that was set out to evaporate in a fume hood. After several days, the water had completely evaporated leaving behind large elongated (>1\u2005cm) green slab-shaped crystals, 2.57\u2005g of which were collected by hand from the dish. These were identified as (I)3)2.6H2O (Aldrich) in 50\u2005ml of water. The resulting red solution was stirred for 30 minutes, transferred to a porcelain dish, and set out to evaporate. The final red product was primarily polycrystalline but some small red\u2013pink plates were found to be suitable for single-crystal X-ray analysis, leading to their identification as (II)A 2.54\u2005g (10.0\u2005mmol) sample of 5-sulfosalicylic acid (3-carb\u00adoxy-4-hy\u00addroxy\u00adbenzene\u00adsulfonic acid) was dissolved in 100\u2005ml of water. To this colorless solution was added a green solution of 2.91\u2005g (10.0\u2005mmol) of Ni(NO6.Uiso constrained to be 1.2 times the Ueq of the bonding atom. Oxygen-bound hydrogen atoms were located in difference electron-density maps and refined with isotropic displacement parameters while the O\u2014H distances were restrained to 0.84\u2005(1)\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022008295/mw2190sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989022008295/mw2190Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989022008295/mw2190IIsup3.hklStructure factors: contains datablock(s) II. DOI: 2202460, 2202459CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure of the title compound, the dimesitylboron group acts to reduce the delocalization of the nitro\u00adgen atom\u2019s lone pair into the pyrrole ring, with increases in the two N\u2014C bond lengths compared to pyrrole itself. The N\u2014B bond is 1.44125\u2005(15)\u2005\u00c5 long. 22H26BN, the B atom acts to reduce the delocalization of the nitro\u00adgen lone-pair electron density into the pyrrole ring, so that the two N\u2014C bonds increase in length to 1.4005\u2005(14) and 1.3981\u2005(14)\u2005\u00c5. The N\u2014B bond length is 1.4425\u2005(15)\u2005\u00c5, which is longer than a typical N\u2014B bond because the nitro\u00adgen lone pair is not fully available to participate in the bond.In the crystal structure of the title compound, C This mol\u00adecule has been investigated previously for its fluorescence, and shows a large Stokes shift, which involves twisted intra\u00admolecular charge transfer (TICT) of the excited state, by rotation about the N\u2014B bond and 3.736\u2005(3)\u2005\u00c5], and is supported by the widening of the C\u2014B\u2014C bond angle [125.18\u2005(9)\u00b0], compared to the two N\u2014B\u2014C bond angles [118.30\u2005(10) and 116.42\u2005(9)\u00b0]. The mesityl groups\u2019 planes lie at 77.14\u2005(4)\u00b0 to each other, and at 69.18\u2005(3) and 67.06\u2005(4)\u00b0 to the pyrrole ring\u2019s best plane. The hydrogen atoms of three methyl groups were modelled in two orientations. The positions and displacement parameters of hydrogen atoms on the pyrrole ring were refined, and for those attached to the C\u03b1 atoms, the H\u2014C\u03b1\u2014C\u03b2 angle showed widening to 131\u2013132\u00b0, similar to that in pyrrole (pyrrol-1-yl)borane ca 0.04\u2005\u00c5 longer than in similar compounds where the nitro\u00adgen atom is attached to two sp3 carbon atoms and where the nitro\u00adgen lone pair is fully available for donation to boron. Compared to the mol\u00adecular geometry of pyrrole itself, as determined by X-ray crystallography at 103\u2005K and 1.3514\u2005(17)\u2005\u00c5 and the C\u03b2\u2014C\u03b2 bond length is 1.4290\u2005(17)\u2005\u00c5. Thus, in contrast to pyrrole, the N\u2014C\u03b1 and C\u03b1\u2014C\u03b2 bonds are no longer similar in length, due to a reduction in the contribution of the nitro\u00adgen atom\u2019s lone pair to the electronic \u03c0 system of the pyrrole ring. When the mesityl groups are replaced by penta\u00adfluoro\u00adphenyl groups in derivative 2, the N\u2014B bond is considerably shorter than in 1 [1.4094\u2005(9) cf. 1.4425\u2005(15)\u2005\u00c5] due to greater lone-pair donation from nitro\u00adgen towards the more electron-deficient boron. Consequently, compared to 1, the pyrrole ring shows slightly longer N\u2014C bonds [1.4033\u2005(6)\u2005\u00c5] and a longer C\u03b2\u2014C\u03b2 bond [1.4418\u2005(9)\u2005\u00c5], though similar lengths for the C\u03b1\u2014C\u03b2 bonds [1.3553\u2005(6)\u2005\u00c5] (Table\u00a011 is similar to that when the pyrrole nitro\u00adgen atom is substituted with a carbonyl group to form an amide (Table\u00a01The N\u2014B bond is 1.4425\u2005(15)\u2005\u00c5 long. This is ] Table\u00a01. For come Table\u00a01.3.ab plane . Within a layer, the mol\u00adecules are related by centres of symmetry and translations along a and b. Adjacent layers are related by the twofold screw and n-glide planes. The four shortest inter\u00admolecular C\u22efH distances are in the range 2.81\u20132.83\u2005\u00c5. Two of these involve the meta-C atom, C18, with a methyl hydrogen atom and a pyrrole ring\u2019s hydrogen atom, which are directed to opposite sides of the phenyl ring [C18\u22efH12A and C18\u22efH3]. The two others involve both a meta and a para-C atom of the second phenyl ring and the pyrrole hydrogen atom H2 [C8\u22efH2 and C9\u22efH2].The mol\u00adecules are packed in layers in the ne Fig.\u00a02. There a4.1 bearing two 2\u2032-thienyl groups in the pyrrole\u2019s 2- and 5-positions \u20131.457\u2005(3)\u2005\u00c5] with a correlation between the increasing angle between bonding planes and longer N\u2014B bonds and 1.440\u2005(3)\u2005\u00c5, similar to those in 1 using hexa\u00adne:di\u00adchloro\u00admethane (8:1) as eluent to give 1 as a slightly oily white solid, from which crystals were grown using ethyl acetate, m.p. 411\u2005K. 1H NMR [ppm]: \u03b4 = 6.84 , 6.81 , 6.37 , 2.32 , 2.11 ; 13C NMR [ppm]: \u03b4 = 141.7 , 139.0 (2 \u00d7 4\u2032-C), 136.5 br (2 \u00d7 1\u2032-C), 128.3 , 126.5 , 114.6 , 22.8 , 21.5 (2 \u00d7 4\u2032-CH3); IR (ATR): 2920, 2853, 1606, 1472, 1451, 1421, 1399, 1378, 1329, 1310, 1287, 1252, 1156, 1122, 1080, 1074, 1043, 1030, 850, 817, 763, 733, 717, 677, 656, 619, 560, 516\u2005cm\u22121.A solution of pyrrole in THF (10\u2005mL) under nitro\u00adgen was treated with sodium hydride and the mixture stirred at 293\u2005K for 2\u2005h. Dimesitylboron fluoride (CARE: gives HF with moisture) in dry THF (10\u2005mL) was added at room temperature. The mixture was left to stir overnight. The bright orange\u2013yellow mixture was quenched with water (20\u2005mL), extracted with ether (2 \u00d7 30\u2005mL) and the combined organic phase was dried over MgSO6.Ueq of the parent atom for aromatic groups and at 1.5Ueq for methyl groups.Crystal data and details of data collection and structure refinement are summarized in Table\u00a0210.1107/S2056989022011768/zv2023sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022011768/zv2023Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022011768/zv2023sup3.docxThis is NMR data requested by the referee. DOI: Click here for additional data file.10.1107/S2056989022011768/zv2023Isup4.cmlSupporting information file. DOI: 2166138CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Due to a production error, there were inaccuracies in the section numbering. The corrected subsection numbering should read as follows:1. Introduction2. Immunotheraputic Strategies Targeting the TME\u20032.1 Therapeutic Strategies Based on TME Inhibitory Signaling\u2003\u20032.1.1 Targeting TME Physical Barriers\u2003\u20032.1.2 Targeting Immune Checkpoints\u2003\u20032.1.3 Targeting Immunosuppresive Cells\u2003\u20032.1.4 Targeting Inhibitory Cytokines\u2003\u20032.1.5 Targeting Metabolic Inhibition Signaling\u20032.2 Therapeutic Strategies Based on TME Stimulatory Signals\u2003\u20032.2.1 Targeting Stimulatory Checkpoints\u2003\u20032.2.2 Application of Stimulating Cytokines\u2003\u20032.2.3 Enhancing Antigen Presentation\u2003\u20032.2.4 Application of Immune Effector cells3. Challenges Faced by Time Research and Solutions\u20033.1 Complexity of TIME\u20033.2 Spatiotemporal Heterogenity of TIME\u20033.3 Systemic Immunity Affects TME Immune Response4. Summary and ProspectThe publisher apologizes for this mistake.The original version of this article has been updated."} +{"text": "MAPK can underlie the pro-differentiation effect of CP-673451 on GBM cells. Overall, the present study identifies a potential novel therapeutic option that could benefit GBM patients in the future, through differentiation of residual GSCs post-surgery, with the aim to limit recurrence and improve quality of life.Glioblastoma (GBM) is the most common and fatal primary brain tumour in adults. Considering that resistance to current therapies leads to limited response in patients, new therapeutic options are urgently needed. In recent years, differentiation therapy has been proposed as an alternative for GBM treatment, with the aim of bringing cancer cells into a post-mitotic/differentiated state, ultimately limiting tumour growth. As an integral component of cancer development and regulation of differentiation processes, kinases are potential targets of differentiation therapies. The present study describes how the screening of a panel of kinase inhibitors (KIs) identified PDGF-R\u03b1/\u03b2 inhibitor CP-673451 as a potential differentiation agent in GBM. We show that targeting PDGF-R\u03b1/\u03b2 with CP-673451 in vitro triggers outgrowth of neurite-like processes in GBM cell lines and GBM stem cells (GSCs), suggesting differentiation into neural-like cells, while reducing proliferation and invasion in 3D hyaluronic acid hydrogels. In addition, we report that treatment with CP-673451 improves the anti-tumour effects of temozolomide in vivo using a subcutaneous xenograft mouse model. RNA sequencing and follow-up proteomic analysis revealed that upregulation of phosphatase DUSP1 and consecutive downregulation of phosphorylated-p38 Glioblastoma (GBM) is the most common and fatal primary brain tumour in adults , 2 with While numerous therapeutic approaches aim for cancer cell death, a few studies have instead started exploring treatments that induce cancer cell differentiation into non-neoplastic cells as a novel strategy . Indeed,GBM treatment could highly benefit from differentiation therapies. High resistance to classic cytotoxic treatments and the presence of pluripotent cancer stem cells (CSCs) in residual tumour tissues due to incomplete surgical excision make GBM a good candidate for differentiation therapies \u201316. SeveIn accordance with these reports, the present study describes the screening of a panel of KIs that led to the identification of a receptor tyrosine kinase (RTK) inhibitor, namely CP-673451, which targets the platelet-derived growth factor receptor \u03b1/\u03b2 (PDGF-R\u03b1/\u03b2) as a potential differentiation agent in GBM. Herein, we propose that CP-673451 treatment can induce the upregulation of phosphatase dual-specificity phosphatase 1 (DUSP1) and consecutive downregulation of phosphorylated p38 mitogen-activated protein kinase (MAPK), leading to the onset of differentiation of GBM cells, thus limiting proliferation and invasion in vitro, while improving the anti-tumour effects of temozolomide in vivo.50, no previously identified undesirable effects at the required concentrations and with the ability to cross the blood-brain barrier (BBB) [p-value) can be found in Table Forty-seven KIs were selected from the KI library (SelleckChem) on the basis of high potency with low ICer (BBB) \u201327. A fun\u2009=\u200947) for 24\u2009h before nine images were taken per well. All images were analysed for total neurite-like process length and normalised by cell number to obtain an average neurite-like process length per cell , pink, blue and red triangles indicate KIs with significant effects compared to the control . Inhibitors showing effects with p\u2009\u2264\u20090.001 were selected for further experiments, namely: LY2835219, a potent and selective inhibitor of CDK4/6 [Neurite-like process outgrowth was quantified following treatment to select KIs with potential differentiating effects. A schematic diagram illustrates the experimental design Fig. with U87ell Fig. . The gref CDK4/6 , CP-6734f CDK4/6 and pacrf CDK4/6 Fig. 1Bn\u2009=\u200947 fop\u2009\u2264\u20090.001), 30.5\u2009\u00b1\u20095.1\u2009\u03bcm (p\u2009\u2264\u20090.01), 52.8\u2009\u00b1\u20096.9\u2009\u03bcm (p\u2009\u2264\u20090.001) and 32.1\u2009\u00b1\u20093.6\u2009\u03bcm (p\u2009\u2264\u20090.01), respectively. In AS, a significant increase of the average neurite-like process length per cell was observed following treatment with CP-673451 (72.1\u2009\u00b1\u200924.63\u2009\u03bcm (p\u2009\u2264\u20090.05)) as compared to the control (28.2\u2009\u00b1\u20095.3\u2009\u03bcm). CP-673451 similarly caused the average neurite-like process length per cell to increase significantly in LN229 cells (25.3\u2009\u00b1\u20097.7\u2009\u03bcm (p\u2009\u2264\u20090.01)) as compared to the control (0.9\u2009\u00b1\u20090.5\u2009\u03bcm). Finally, LY2835219, pelitinib and CP-673451 treatments caused a significant increase in the average neurite-like process length (15.3\u2009\u00b1\u20095.2\u2009\u03bcm (p\u2009\u2264\u20090.05), 15.9\u2009\u00b1\u20096.4\u2009\u03bcm (p\u2009\u2264\u20090.05) and 56.0\u2009\u00b1\u20099.6 (p\u2009\u2264\u20090.001) respectively) in the U138 cells compared to the control (3.9\u2009\u00b1\u20090.3\u2009\u03bcm) , as a normal brain cell type control . Neurite\u03bcm) Fig. .Fig. 2Sep\u2009\u2264\u20090.01) compared to the control, while a slight decrease in viability was observed in U138 and LN229 GBM cells following treatment with pacritinib (0.9\u2009\u00b1\u20090.05\u2009RAU (p\u2009\u2264\u20090.05) and 0.8\u2009\u00b1\u20090.06\u2009RAU (p\u2009\u2264\u20090.05), respectively). Treatments with LY2835219, pelitinib and pacritinib were also observed to significantly affect LN229 cell proliferation compared to the control (0.8\u2009\u00b1\u20090.09\u2009RAU (p\u2009\u2264\u20090.01), 0.8\u2009\u00b1\u20090.1\u2009RAU (p\u2009\u2264\u20090.05) 0.8\u2009\u00b1\u20090.1\u2009RAU (p\u2009\u2264\u20090.05), respectively). Altogether, treatment with 1\u2009\u03bcM of the selected KIs appeared to stimulate neurite-like process outgrowth in most studied GBM cell lines and astrocytes, with limited impact on cell viability and proliferation.To examine how various treatments affected cell viability and growth, metabolic activity and proliferation were measured following treatment with the inhibitors of interest using WST-1 ) in G166 cells as compared to control (1.2\u2009\u00b1\u20090.6\u2009\u03bcm). In addition, 1\u2009\u00b5M LY2835219, CP-673451 and pacritinib led to significant increases in the average neurite-like process length (35.9\u2009\u00b1\u20097.0\u2009\u03bcm (p\u2009\u2264\u20090.01), 21.6\u2009\u00b1\u20092.7\u2009\u03bcm (p\u2009\u2264\u20090.01) and 20.3\u2009\u00b1\u20095.1\u2009\u03bcm (p\u2009\u2264\u20090.05), respectively) in GS090 GSCs compared to the control (13.8\u2009\u00b1\u20092.5\u2009\u03bcm). Finally, 1\u2009\u00b5M LY2835219, pelitinib and CP-673451 significantly increased the average neurite-like process length per cell (26.7\u2009\u00b1\u20096.0\u2009\u03bcm (p\u2009\u2264\u20090.05) 25.2\u2009\u00b1\u20093.2\u2009\u03bcm (p\u2009\u2264\u20090.05) and 28.1\u2009\u00b1\u20094.6\u2009\u03bcm (p\u2009\u2264\u20090.05), respectively) in G179 GSCs, compared to the control (12.3\u2009\u00b1\u20094.4\u2009\u03bcm). Representative light microscopic images are shown, with neurite-like processes indicated by red arrows.As one of the drivers of GBM recurrence with multipotent abilities, GSCs are the putative main targets of differentiation therapy. For these reasons, the selected KIs were used to treat different GSC lines, including G166, GS090 and G179. Their morphology was observed using light microscopy following treatment and the average neurite-like process length per cell was calculated as before Fig. . Treatmep\u2009\u2264\u20090.05) and 0.9\u2009\u00b1\u20090.07\u2009RAU (p\u2009\u2264\u20090.05), respectively). On the other hand, cell viability increased significantly following treatment with CP-673451 (1.04\u2009\u00b1\u20090.02\u2009RAU (p\u2009\u2264\u20090.05)) as compared to the control. Further, proliferation assays indicated that G166 cell proliferation decreased after pelitinib treatment (0.9\u2009\u00b1\u20090.04\u2009RAU (p\u2009\u2264\u20090.01)) but increased upon treatment with CP-673451 (1.3\u2009\u00b1\u20090.14\u2009RAU (p\u2009\u2264\u20090.05)) as compared to the control. Regarding GS090 cells, CP-673451 treatment decreased proliferation (0.99\u2009\u00b1\u20090.01\u2009RAU (p\u2009\u2264\u20090.01)) as compared to the control. None of the KI treatments caused significant changes in G179 proliferation.GSC viability (WST-1) -PDGF-R\u03b1/\u03b2 (tyr849/tyr857) was measured as well as PDGF-R\u03b2 upon treatment with CP-673451. U87 GBM cells were treated with 1\u2009\u00b5M CP-673451 for 0, 15\u2009min, 1, 4, 24 and 48\u2009h ), PROM-1 (1.64\u2009\u00b1\u20090.18\u2009fc (p\u2009\u2264\u20090.01)) and VEGFA (1.29\u2009\u00b1\u20090.05\u2009fc (p\u2009\u2264\u20090.001)). Treatment also indicated significant decreases in GFAP (0.54\u2009\u00b1\u20090.07\u2009fc (p\u2009\u2264\u20090.001)), CSPG4 (0.57\u2009\u00b1\u20090.04\u2009fc (p\u2009\u2264\u20090.0001)) and CD44 (0.7\u2009\u00b1\u20090.03\u2009fc (p\u2009\u2264\u20090.001)). Upon treatment with 1\u2009\u03bcM CP-673451, G179 GSCs also showed a significant increase in the expression of NEFL (3.47\u2009\u00b1\u20090.51\u2009fc (p\u2009\u2264\u20090.0001) and significant decreases in the expression of nestin and PROM1 (0.69\u2009\u00b1\u20090.09\u2009fc (p\u2009\u2264\u20090.001) and 0.18\u2009\u00b1\u20090.37\u2009fc (p\u2009\u2264\u20090.01), respectively), CSPG4 (0.65\u2009\u00b1\u20090.06\u2009fc (p\u2009\u2264\u20090.001) and VEGFA (0.66\u2009\u00b1\u20090.03\u2009fc (p\u2009\u2264\u20090.001).Gene expression of specific differentiation markers was quantified using qRT-PCR in order to determine which specific normal brain cell lineage the GBM cells were differentiating into upon CP-673451 treatment. The selected differentiation markers were as follows; Astrocytic (GFAP and ALDH1L1), Neuronal (\u03b23-tubulin and NEFL), stem cell (nestin and PROM-1 (CD133), oligodendrocyte (OLIG2 and CSPG4) and mesenchymal (VEGFA and CD44). Upon treatment of U87 GBM cells with 1\u2009\u00b5M CP-673451 for 96\u2009h Fig. , cells sAltogether our data showed that treatment with CP-673451 could trigger a neuron-like differentiation in GBM cells, as supported by the formation of neurite-like processes and expression of the neuronal NEFL marker in GBM cells, with concomitant decrease of expression of markers for stem cells and other brain cell types.p\u2009\u2264\u20090.05)). Conversely, a significant decrease in cell proliferation was observed in GS090 cells upon treatment with 5\u2009\u00b5M CP-673451 (0.7\u2009\u00b1\u20090.03\u2009RAU (p\u2009\u2264\u20090.01)), as compared to the control. All three cell lines showed a significant decrease in proliferation when treated with 10\u2009\u00b5M CP-673451: U87 (0.7\u2009\u00b1\u20090.04\u2009RAU (p\u2009\u2264\u20090.001)), GS090 (0.6\u2009\u00b1\u20090.07\u2009RAU (p\u2009\u2264\u20090.001)) and G179 (0.7\u2009\u00b1\u20090.06\u2009RAU (p\u2009\u2264\u20090.01)), as compared to the control.To further characterise the effect of CP-673451 on proliferation, additional experiments with increasing concentrations of the inhibitor were performed. The U87 GBM cell line and GS090 and G179 GSCs were treated with increasing concentrations of CP-673451 for 48\u2009h, while a proliferation assay was also performed Fig. . A signip\u2009\u2264\u20090.001) and G179: 0.6\u2009\u00b1\u20090.07 (p\u2009\u2264\u20090.001). Following CP-673451 treatment, significant decreases in cell colony size were observed at 1, 5 and 10\u2009\u00b5M in U138 (0.7\u2009\u00b1\u20090.17 (p\u2009\u2264\u20090.05), 0.6\u2009\u00b1\u20090.11 (p\u2009\u2264\u20090.01), 0.5\u2009\u00b1\u20090.05 (p\u2009\u2264\u20090.0001)) and GS090 (0.8\u2009\u00b1\u20090.09 (p\u2009\u2264\u20090.05) 0.7\u2009\u00b1\u20090.08 (p\u2009\u2264\u20090.01) and 0.5\u2009\u00b1\u20090.1 (p\u2009\u2264\u20090.01)). No significant alterations of the number of colonies were observed in the tested cell lines upon CP-673451 treatment. Yet, a significant decrease in U87 cell metabolic activity and 0.8\u2009\u00b1\u20090.06 (p\u2009\u2264\u20090.01), respectively). Similarly, U138 showed a significant decrease in metabolic activity upon treatment with 1, 5 and 10\u2009\u03bcM CP-673451 (0.7\u2009\u00b1\u20090.05 (p\u2009\u2264\u20090.001), 0.7\u2009\u00b1\u20090.07 (p\u2009\u2264\u20090.01), 0.7\u2009\u00b1\u20090.06 (p\u2009\u2264\u20090.001)), while both GS090 and G179 GSCs showed a significant decrease in metabolic activity upon CP-673451 treatment at 10\u2009\u00b5M (0.6\u2009\u00b1\u20090.11 (p\u2009\u2264\u20090.01)) and (0.9\u2009\u00b1\u20090.04 (p\u2009\u2264\u20090.01)), respectively. Altogether, it appears that treatment with CP-673451 limits the proliferation and invasiveness of GBM cells while stimulating their neuron-like differentiation in vitro.A HA based hydrogel assay was implemented to assess the invasiveness and colony forming ability of the U87 and U138 GBM cell lines and GS090 and G179 GSCs in a 3D model when treated with increasing concentrations of CP-673451 . Cell metabolic activity was also measured in parallel (using a CellTiter-Glo luminescent cell viability assay). Both the colony areas Fig. and coloity Fig. was measIn the interest of ensuring the effects from pharmacological inhibition in U87 GBM cells upon CP-673451 treatment were due to specific inhibition of its targets PDGF-R\u03b1 and PDGF-R\u03b2, gene silencing using siRNA was performed.p\u2009\u2264\u20090.0001) and 95% 0.05\u2009\u00b1\u20090.03 (p\u2009\u2264\u20090.0001), respectively. In addition, PDGF-R\u03b2 expression was also reduced at both 50\u2009nM and 100\u2009nM by 47% 0.53\u2009\u00b1\u20090.18 (p\u2009\u2264\u20090.01) and 46% 0.53\u2009\u00b1\u20090.23 (p\u2009\u2264\u20090.05), respectively. KD of PDGF-R\u03b1 and \u03b2 was also validated in G179 cells ) and PDGF-R\u03b2 48% (0.52\u2009\u00b1\u20090.2 (p\u2009\u2264\u20090.01)) compared to the control. At 100\u2009nM siRNA, the gene expression of PDGF-R\u03b1 was reduced by 57% (0.43\u2009\u00b1\u20090.1 (p\u2009\u2264\u20090.0001)) while gene expression of PDGF-R\u03b2 was reduced by 48% (0.52\u2009\u00b1\u20090.02 (p\u2009\u2264\u20090.001)). Further validating the siRNA transfection, the protein expression of PDGF-R\u03b1 and PDGF-R\u03b2 was decreased in U87 GBM cells and G179 GSCs. PDGF-R\u03b2 protein expression was reduced by 72% in U87 GBM cells at both 50\u2009nM and 100\u2009nM concentrations of siRNA compared to control (0.28\u2009\u00b1\u20090.1 (p\u2009\u2264\u20090.0001)) and PDGF-R\u03b2 by 33% 0.67\u2009\u00b1\u20090.2 (p\u2009\u2264\u20090.05). 100\u2009nM siRNA reduced PDGF-R\u03b1 protein expression by 26% 0.74\u2009\u00b1\u20090.1 (p\u2009\u2264\u20090.05) and PDGF-R\u03b2 by 40% 0.60\u2009\u00b1\u20090.2 (p\u2009\u2264\u20090.05) ). Treatment of U87 GBM cells with 100\u2009nM PDGF-R\u03b2 siRNA compared to the control (18.9\u2009\u00b1\u20091.7\u2009\u00b5m) displayed a significant increase in neurite-like process length average per cell (29.8\u2009\u00b1\u20092.9\u2009\u00b5m (p\u2009\u2264\u20090.001)), as did the combination of PDGF-R\u03b1 and \u03b2 siRNA (29.1\u2009\u00b1\u20096.8\u2009\u00b5m (p\u2009\u2264\u20090.05)). Similarly, KD PDGF-R\u03b1 in G179 cells with siRNA at 50 and 100\u2009nM significantly increased the average length of neurite-like process per cell (10.7\u2009\u00b5m\u2009\u00b1\u20092.1 (p\u2009\u2264\u20090.05)) compared to the control (7.9\u2009\u00b1\u20091.5\u2009\u00b5m) and (12.4\u2009\u00b1\u20092.9\u2009\u00b5m (p\u2009<\u20090.05)) compared to the control (8.8\u2009\u00b1\u20091.3\u2009\u00b5m), respectively. KD of both PDGF-R\u03b1 and \u03b2 also increased neurite-like process outgrowth compared to the controls (10.7\u2009\u00b1\u20091.2\u2009\u00b5m (p\u2009\u2264\u20090.05)) ) Fig. .Fig. 4GeAs specific gene silencing of PDGF-R\u03b1 and PDGF-R\u03b2 enhanced neurite-like process growth in U87 GBM cells and G179 GSCs, these data suggest that the observed impact of CP-673451 treatment on GBM cells should be, at least partly, due to specific PDGF-R\u03b1/\u03b2 inhibition.3): TMZ: 16.75\u2009\u00b1\u20095.4\u2009mm3 (p\u2009\u2264\u20090.01), CP-673451: 35.9\u2009\u00b1\u200916.9\u2009mm3 (p\u2009\u2264\u20090.01) and TMZ\u2009+\u2009CP-673451: 6.06\u2009\u00b1\u20092.3 mm3 (p\u2009\u2264\u20090.01). Tumour volumes from the combination treatment group were significantly reduced compared to both singular treatments.The in vivo anti-tumour activity of CP-673451 was evaluated using a U87 xenograft GBM mouse model Fig. . Treatmep\u2009\u2264\u20090.01), CP-673451: 352.4\u2009\u00b1\u2009115.6 (p\u2009\u2264\u20090.05) and TMZ\u2009+\u2009CP-673451: 36.2\u2009\u00b1\u200912.1 (p\u2009\u2264\u20090.001). There were significantly fewer ki67-positive cells in the TMZ\u2009+\u2009CP-673451 group compared to CP-673451 treatment alone. Altogether it appeared that CP-673451 treatment can improve the inhibitory effect of TMZ on tumour growth in vivo.Tumour samples were stained for proliferation marker Ki67 Fig. . The numTo decipher the signalling mechanism underlying the effect of CP-673451 on GBM cell differentiation, RNA-seq was performed on U87 GBM cells treated with CP-673451 for 48\u2009h (compared to DMSO control). Whole transcriptome correlation matrix showed a high similarity between the replicate samples Fig. . DESEQ2 p-value\u2009=\u20090.003) and Hallmark NF-\u03baB signalling in comparison to U87 GBM cells treated with DMSO in CP-673451 treated U87 GBM cells compared to control [p\u2009\u2264\u20090.0001), 2.5\u2009\u00b1\u20091.0 (ns), respectively) ) and 45% (0.55\u2009\u00b1\u20090.14\u2009fc (p\u2009\u2264\u20090.001)) compared to the control siRNA (siCTRL) simultaneously at 50\u2009nM for 24\u2009h. G179 GSCs were treated by reverse transfection with 100\u2009nM DUSP1 siRNA (SMPs 3 and 4) simultaneously for 48\u2009h. DUSP1 KD was validated by qRT-PCR in U87 GBM cells and G179 GSCs 62% (0.38\u2009\u00b1\u20090.04\u2009fc (vs 53.8\u2009\u00b1\u20098.27\u2009\u03bcm (p\u2009\u2264\u20090.01)) and G179 GSCs (30% (13.0\u2009\u00b1\u20091.1\u2009\u00b5m vs 18.5\u2009\u00b1\u20092.2\u2009\u00b5m (p\u2009\u2264\u20090.01)) ) and 21% (14.2\u2009\u00b1\u20090.8\u2009\u00b5m vs 18.0\u2009\u00b1\u20092.3\u2009\u03bcm (p\u2009\u2264\u20090.05)), respectively ) Fig. . In addiely Fig. .Fig. 7DUp\u2009\u2264\u20090.05)) and G179 GSCs (3.7\u2009\u00b1\u20091.4 fold increase (p\u2009\u2264\u20090.05)) compared to control fold in U87 GBM cells and 30.8\u2009\u00b1\u200913.1 fold (p\u2009\u2264\u20090.05) in G179 GSCs compared to control . In G179 GSCs, transfection with the DUSP1 plasmid increased the average neurite-like process length by 47% compared to the control 7.9\u2009\u00b1\u20090.3\u2009\u00b5m vs 5.4\u2009\u00b1\u20090.6\u2009\u03bcm (p\u2009\u2264\u20090.01) and 0.72\u2009\u00b1\u20090.16 (p\u2009\u2264\u20090.05), respectively). Co-treatment with DUSP1 inhibitor BCI abrogated this decrease in p38MAPK phosphorylation is essential to the growing neurites of the developing neurons , 35, 36.MAPK, one of DUSP1 down-stream targets, also including p44/p42MAPK (ERK1/2) and SAPK/JNK [via p38MAPK inhibition due to DUSP1 activity. Interestingly, a separate study reported that high DUSP1 levels correlate with increased GBM patient survival. Authors revealed that overexpression of DUSP1 in GSCs impedes self-renewal and induces differentiation via deactivation of p38MAPK in vitro, reducing tumourigenicity and increasing sensitivity to TMZ therapy [Further supporting this observation, our RNA-seq analysis identified DUSP1 expression to be significantly upregulated in GBM cells upon treatment with CP-673451. DUSP1 has been reported to be essential in the early hours of neuronal differentiation during embryogenesis. Its activity is required for both limiting cell proliferation and ensuring proper neurite outgrowth. DUSP1 is also directly implicated in the maintenance of neuronal integrity . PreviouSAPK/JNK , 38, 41.Taken together, these results thus suggest that CP-673451 treatment could hold great promise as part of a novel therapeutic strategy against GBM. CP-673451 is a low molecular weight kinase inhibitor (molecular weight\u2009=\u2009417.52) with thevia DUSP1 is being reported. Yet, our present investigation presents some limitations that would need to be addressed in follow-up studies. Among these, the upstream mechanism between PDGF-R inhibition and upregulation of DUSP1 is yet to be elucidated. However, our RNA-seq data demonstrated a significant enrichment of genes linked to the NF-\u03baB pathway upon treatment of GBM cells with CP-673451. Accordingly, the promoter region of the DUSP1 gene contains binding sites for NF-\u03baB [To our knowledge, this is the first time such an effect of PDGF-R inhibition on GBM cell differentiation or NF-\u03baB . In addior NF-\u03baB , 45. Lasor NF-\u03baB , 51.Altogether, this study has identified a KI, CP-673451, able to induce differentiation in GBM cells, with the potential to target GSCs, which are known to be directly implicated in GBM therapeutic resistance and inevitable recurrence. CP-673451 treatment could thus refine therapeutic strategies against GBM, through reducing side effects and enhancing response to current therapies, consequently improving patients\u2019 quality of life.2 atmosphere. Astrocytes and established human GBM cell lines were purchased from the American Type Culture Collection (ATCC). Astrocytes (AS) were seeded onto flasks and plates pre-treated with 2\u2009\u03bcg/cm2 poly-L-lysine (Sigma-Aldrich) and required Astrocyte growth medium (Sigma-Aldrich) 10% foetal bovine serum (FBS) (Sigma-Aldrich) and 100 units ml\u22121 penicillin, 100\u2009\u03bcg\u2009ml\u22121 streptomycin and L-glutamine (1% PSG) (Sigma-Aldrich). U87 and U138 cells required minimum essential medium (MEM) 10% FBS and 1% PSG. LN229 cells required Dulbecco\u2019s modified eagle medium (DMEM) 10% FBS and 1% PSG.Cells were maintained at 37\u2009\u00b0C in a humidified 5% COPatient-derived GSCs were a kind gift from Dr. Angela Bentivegna, University of Milan-Bicocca, Italy and Dr. David Nathanson, University of California, Los Angeles, US. GSCs were isolated from GBM tumour samples following local ethical board approval. GSCs were maintained as neurospheres in DMEM/F12 Ham (Sigma-Aldrich), 1% B27 without vitamin A (Gibco), 1% Glutamax (Gibco) and 1% PSG with a mix of growth factors . Medium was changed twice a week. GBM cell lines and astrocytes were detached at confluence using trypsin/EDTA (Sigma-Aldrich). GSCs were disassociated using TrypleE express enzyme (Gibco) and separated into single cells through a 70\u2009\u03bcm cell strainer (Starstedt). All cells were tested for mycoplasma at the beginning of the study.3 cells per well) into a 96-well plate and treated with 1\u2009\u03bcM of 47 KIs from a library of 378 (SelleckChem) for 24\u2009h in 10% FBS. The automated liquid handler Biomek 4000 was used for performing this screening (Beckman Coulter). Dimethyl sulfoxide (DMSO) was used as a vehicle control. GSCs were treated with 10% FBS to enhance cell attachment. Following 24\u2009h (GBM cell lines) and 48\u2009h (GSCs) treatment with KIs, images of the cells were taken, and length of neurite-like process was measured as described below.Cells were counted using the automated cell counter Countess (Invitrogen), seeded (2\u2009\u00d7\u200910Light microscopic images were taken at 20x magnification (mag) using the Olympus IX71 microscope with Micromanager software. The length of long thin extensions termed \u2018neurite-like processes\u2019 were analysed. Three technical repeats were performed with nine images taken per well and all neurite-like processes analysed. The semi-automatic Fiji (ImageJ) plugin \u2018simple neurite tracer\u2019 was used to measure the lengths of the neurite-like processes. The total lengths of neurite-like processes per well were quantified and normalised by the total number of cells per well to calculate the average length of neurite-like process per cell. Graphs display the mean\u2009\u00b1\u2009SD of at least three independent experiments, representative images shown, red arrows indicating neurite-like processes.3 or 5\u2009\u00d7\u2009103 cells per well) into a 96-well plate with 10% FBS 1% PSG for 24\u2009h before the treatments were applied. Cells were then treated with 0, 1, 5 or 10\u2009\u00b5M of the stated KIs. Following 24\u2009h (GBM cell lines) or 48\u2009h (GSCs) treatment, cells were fixed with 4% paraformaldehyde (PFA) in 1x phosphate buffered saline (PBS) (Sigma-Aldrich) and incubated at room temperature (temp) for 30\u2009min. PFA was removed and plates left to dry at room temp for 10\u2009min. They were then washed once with PBS and stained with 0.1% crystal violet solution at room temp for 30\u2009min. The solution was then aspirated and washed in 1x PBS twice and water once and left open overnight to dry. When ready to read, 50\u2009\u03bcl of 10% acetic acid was added to each well, plates were agitated on a plate rocker for 20\u2009min and absorbance was recorded at 600\u2009nm using a GloMax Explorer reader (Promega). Graphs show the mean\u2009\u00b1\u2009SD of at least three independent experiments.Cells were seeded (2\u2009\u00d7\u2009103 cells per well) into a 96-well plate with 10% FBS 1% PSG for 24\u2009h before the treatments were applied. Cells were then treated with DMSO or 1\u2009\u00b5M of the stated KIs. Following 24\u2009h (GBM cell lines) or 48\u2009h (GSCs) treatment, 10\u2009\u03bcl of water-soluble tetrazolium salts-1 (WST-1) reagent was added to each well and incubated at 37\u2009\u00b0C, 5% CO2 for 2\u2009h. The plate was then agitated on a plate rocker at room temp for 1\u2009min and the absorbance recorded at 450\u2009nm using a GloMax Explorer reader (Promega). Graphs show the mean\u2009\u00b1\u2009SD of at least three independent experiments.Cells were seeded (2\u2009\u00d7\u200910U87 GBM cells were treated with DMSO or CP-673451 (CP) for 48\u2009h and RNA was purified following PureLink RNA Mini Kit (Life technologies) protocol, as described before . All conhttp://hannonlab.cshl.edu/fastx_toolkit/license.html). The quality of reads were confirmed using the fastqc tool kit (v 0.11.5) (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/), and only high-quality clean read were used for down-stream analyses. The high-quality reads were mapped to the ENSEMBL built GRCH37 using the STAR aligner (v2.5.3a) [p value (Padj\u2009\u2264\u20090.05) and Log2 fold change\u2009\u2265\u2009|1| were used as the threshold to screen significance of differentially expressed genes (DEGs). Gene set enrichment function from the enrichplot package (https://github.com/YuLab-SMU/enrichplot) was used to perform gene set enrichment analysis for Hallmark gene sets and CAHOY CNS neural cell type gene sets deposited in the MSIGDB (https://www.gsea-msigdb.org/gsea/msigdb/index.jsp).High-quality clean reads were obtained by trimming the adaptor sequences and removing reads that contained poly-N or were of low-quality from the raw data using the fastX tool kit (v 0.0.14) (v2.5.3a) with thev2.5.3a) . Differe5 cells per T75\u2009cm2 or 1\u2009\u00d7\u2009105 per well into a 6-well plate) and were treated with drug/control or target/control siRNA plasmid/control for the concentration and time stated. Cells were lysed, RNA was then purified following PureLink RNA Mini kit (Life technologies) protocol. Reverse transcription was carried out using a cDNA synthesis kit (Applied Biosystems). Taqman (Applied Biosystems) or SYBR Green (Applied Biosystems) gene expression master mix and synthesised cDNA was mixed with primers (Applied Biosystems) (Tables Cells were seeded (5\u2009\u00d7\u200910) Tables and run 5 cells were seeded per T75cm2 flask and incubated for 24\u2009h before treatment with: 1\u2009\u00b5M CP-673451 for 0, 15\u2009min, 1, 4, 24 and 48\u2009h vs DMSO (control) in 0% FCS medium; 1\u2009\u03bcM CP-673451 and 1\u2009\u03bcM DUSP1 inhibitor BCI ((E)\u22122-benzylidene-3-(cyclohexylamino)\u22122,3-dihydro-1H-inden-1-one) (Merck Millipore) vs 1\u2009\u03bcM BCI and DMSO vs 1\u2009\u03bcM CP-673451 vs DMSO (control) for 24\u2009h in 10% FCS medium. Cells were collected and protein extracted using RIPA buffer (Sigma) including fresh protease and phosphatase inhibitors (Roche). Standard western blotting protocol was performed [MAPK , anti-p38MAPK , anti-phos-SAPK/JNK , anti-SAPK/JNK , anti-phos-p44/p42MAPK , anti-p44/p42MAPK and anti-\u03b2-actin (Genscript #A00702 1:1000 dilution)) were prepared in the same solution used for blocking and incubated on a rocker at 4\u2009\u00b0C overnight (Table 5\u2009\u00d7\u200910erformed with a 15 cells per well) into a 96-well HA hydrogel assay plate. As per instructions, 40\u2009\u00b5l of cell suspension was added as a drop on top of the gel, incubated for 30\u2009min then 100\u2009\u00b5l media was added, as previously described [Cells were seeded . Cells were incubated at room temp for 30\u2009min before 100\u2009\u00b5l CellTiter-Glo reagent was added to each well, agitated on a plate rocker for 2\u2009min and incubated at room temp for 10\u2009min. Luminescence was recorded with an integration time of 0.3\u2009s using a GloMax Explorer reader (Promega). Graphs show the mean\u2009\u00b1\u2009SD of at least three independent experiments.3 (U87 GBM cells) or 5\u2009\u00d7\u2009103 (G179 GSCs) cells per well) into a 96-well plate and incubated at 37\u2009\u00b0C, 5% CO2 for 24\u2009h. Transfection was performed according to the Lipofectamine 3000 (Invitrogen) user guide. Lipofectamine and siRNA complexes were diluted separately in Opti-MEM (Gibco). These solutions were mixed and incubated at room temp for 15\u2009min. Media was then aspirated, fresh media was added to the cells and the mixed siRNA/Lipofectamine/Opti-MEM solution was added dropwise. Cells were transfected for 48\u2009h with 50\u2009nM and 100\u2009nM siRNA (control (CTRL), PDGF-R\u03b1, PDGF-R\u03b2 or a combination of PDGF-R\u03b1 and \u03b2) (Table 5 cells per well) into a 6-well plate, transfected as described with 50\u2009nM and 100\u2009nM CTRL siRNA or a combination of PDGF-R\u03b1 and PDGF-R\u03b2 siRNA. At 48\u2009h cells were lysed, RNA purified, cDNA synthesised and mixed with SYBR Green master mix (Applied Biosystems) plus primers (Table Cells were seeded (2\u2009\u00d7\u200910Plasmid transfection was performed with 1\u2009ng/\u00b5l pCMV6 empty vector or DUSP1 plasmid (Origene RC205220) for 48\u2009h using transfection reagent HiPerFect. Graphs display the mean\u2009\u00b1\u2009SD of at least three independent experiments, representative images shown.6 U87 GBM cells subcutaneously into 6-week-old immunocompromised athymic nude mice . Once tumours were palpable (tumour diameter ~ 3\u2009mm3), mice were randomly divided into the separate groups: DMSO control (n\u2009=\u20096), TMZ (25\u2009mg/kg/day) (n\u2009=\u20099), CP-673451 (40\u2009mg/kg/day) (n\u2009=\u20099) or combination of TMZ (25\u2009mg/kg/day) and CP-673451 (40\u2009mg/kg/day) (n\u2009=\u20099). Treatments were administered by oral gavage 5 days per week for 3 weeks after which the mice were sacrificed, tumours extracted and tumour volumes measured using the formula: (volume (mm3)\u2009=\u2009(length\u2009x\u2009height2)/2). Paraffin-embedded tumours were sectioned at a thickness of 4\u2009mm with five tissue samples obtained per tumour. Samples were placed in a pressure cooker for 15\u201320\u2009min in 0.01\u2009M citrate buffer (pH 6.0) to remove aldehyde links formed during initial fixation of tissues. Specimens were incubated with antibodies specific for Ki-67 (1:100) overnight at 4\u2009\u00b0C and immunodetection performed the following day using 3,3\u2032-diaminobenzidine (DAB) (Dako) according to manufacturer\u2019s instructions. Images were obtained using Olympus BX63 microscope (200x mag) and the number of cells per field quantified.Following local authority ethical approval by Sun Yat-sen University Animal Care and Use Committee, xenograft mouse models were developed by injecting 2\u2009\u00d7\u200910T-tests were used to compare control group with each treatment group. Differences were considered statistically significant at p\u2009\u2264\u20090.05. .Sample size was set to a minimum of three independent experiments and experimental findings were reliably reproducible. GraphPad Prism was used for data analysis and graphs. All data are presented as mean\u2009\u00b1\u2009SD. Supplementary Figures 1\u20135"} +{"text": "The results obtained from crystal packing and DFT analysis suggest that the mol\u00adecules are held together by forces such as hydro\u00adgen bonding and \u03c0\u2013\u03c0 inter\u00adactions.Three o-nitro\u00adsul\u00adfonamides and N-cyclo\u00adamino-o-sul\u00adfan\u00adil\u00adamides were synthesized and characterized using techniques including 1H NMR, 13C NMR and FT\u2013IR spectroscopy, and single-crystal X-ray diffraction (SC-XRD). The calculated density functional theory (DFT)-optimized geometry of the mol\u00adecules showed similar conformations to those obtained by SC-XRD. Mol\u00adecular docking of N-piperidinyl-o-sul\u00adfan\u00adil\u00adamide and N-indolinyl-o-sul\u00adfan\u00adil\u00adamide supports the notion that o-sul\u00adfan\u00adil\u00adamides are able to bind to human carbonic anhydrase II and IX inhibitors . Hirshfeld surface analyses and DFT studies of three o-nitro\u00adsul\u00adfonamides {1-[(2-nitro\u00adphen\u00adyl)sul\u00adfon\u00adyl]pyrrolidine, C10H12N2O4S, 1, 1-[(2-nitro\u00adphen\u00adyl)sul\u00adfon\u00adyl]piperidine, C11H14N2O4S, 2, and 1-[(2-nitro\u00adphen\u00adyl)sul\u00adfon\u00adyl]-2,3-di\u00adhydro-1H-indole, C14H12N2O4S, 3} and three N-cyclo\u00adamino-o-sul\u00adfan\u00adil\u00adamides suggested that forces such as hydro\u00adgen bonding and \u03c0\u2013\u03c0 inter\u00adactions hold mol\u00adecules together and further showed that charge transfer could promote bioactivity and the ability to form biological inter\u00adactions at the piperidinyl and phenyl moieties.In the search for new \u2018sulfa drugs\u2019 with therapeutic properties, Sulfonamides E are derivatives of sul\u00adfan\u00adil\u00adamide and remain an important class of drugs, with anti\u00adbacterial and non-anti\u00adbacterial potencies, such as diuretic, anti\u00admicrobial, anti-epileptic, anti\u00adleprotic, anti\u00admalarial, hypoglycemic, anti\u00adretro\u00adviral, anti\u00adthyroid and anti-inflammatory activities sul\u00adfon\u00adyl]pyrrolidine, 1, 1-[(2-nitro\u00adphen\u00adyl)sul\u00adfon\u00adyl]pi\u00adperi\u00addine, 2, and 1-[(2-nitro\u00adphen\u00adyl)sul\u00adfon\u00adyl]-2,3-di\u00adhydro-1H-indole, 3, and the N-cyclo\u00adamino-o-sul\u00adfan\u00adil\u00adamides 2-(pyrrolidine-1-sul\u00adfon\u00adyl)aniline, 4, 2-(piperidine-1-sul\u00adfon\u00adyl)aniline, 5, and 2-aniline, 6. The crystal structures, density functional theory (DFT) studies, Hirshfeld surface analysis, mol\u00adecular electrostatic potential and electronic properties of the title sul\u00adfonamides and sul\u00adfan\u00adil\u00adamides (1\u20136) have been discussed. Mol\u00adecular docking experiments with carbonic anhydrase II (PDB entry 4iwz) and IX (5fl4) active sites were conducted in order to predict their binding inter\u00adactions with 1\u20136 solvent system visualized under a UV lamp (254\u2005nm). Column chromatography was performed with silica gel (70\u2013230 mesh ASTM) and mobile phases were as indicated. Sample crystallization was achieved by the slow evaporation of the indicated solvent systems at ambient tem\u00adper\u00adature. IR spectra were obtained using a Bruker Tensor 27 platinum ATR\u2013FT\u2013IR spectrometer. The ATR\u2013FT\u2013IR spectra were acquired in a single mode with a resolution of 4\u2005cm\u22121 over 32 scans, in the region 4000\u2013650\u2005cm\u22121. 1H and 13C NMR spectra were recorded, in CDCl3, on a Bruker 400\u2005MHz spectrometer. Chemical shift (\u03b4) values were measured in parts per million (ppm) downfield from tetra\u00admethyl\u00adsilane (TMS) and coupling constants (J) are reported in hertz (Hz). Theoretical studies were performed for the com\u00adpounds and, in each case, their SC-XRD structures were used for optimization and global reactivity descriptor (GRD) calculations.All reagents were purchased from Millipore Sigma (Ger\u00admany and South Africa) and were used without purification. The melting points were determined on an Electrothermal digital melting-point apparatus and are uncorrected. Reactions were monitored by thin-layer chromatography (TLC) on Merck silica gel 60 Fo-Nitro\u00adbenzene\u00adsul\u00adfonyl chloride (1.00\u2005mmol) was added slowly to a stirring dried toluene solution (30\u2005ml) of the cyclo\u00adamine (2.20\u2005mmol) at ambient tem\u00adper\u00adature and stirred for 12\u2005h, monitored by TLC. The reaction mixture was then diluted with di\u00adchloro\u00admethane (30\u2005ml) and washed with distilled water (3 \u00d7 10\u2005ml). The organic layer was separated, dried over anhydrous sodium sulfate, filtered and concentrated to an oil, which was purified by column chromatography on silica gel . Crystals were obtained by the slow solvent evaporation of the requisite eluates at ambient tem\u00adper\u00adature, except for 5, which was recrystallized from di\u00adchloro\u00admethane, slowly evaporated and filtered to give single crystals.1. o2.2.1.1. N-Pyrrolidinyl-o-nitro\u00adbenzene\u00adsul\u00adfonamide, -Nitro\u00adbenzene\u00adsul\u00adfonyl chloride and pyrrolidine . Yellow crystals ; RF\u00a0= 0.44 ; m.p. 81.7\u201381.9\u2005\u00b0C. IR : 3080 (aryl C\u2014H str.), 2968 (sp3-C\u2014H str.), 1597 (aryl C=C str.), 1543 (asym C\u2014NO2 str.), 1344 (sym C\u2014NO2 str.), 1342 (asym SO2\u2014N str.), 1163 (sym SO2\u2014N str.), 1078 (C\u2014N str.). 1H NMR : 7.94 , 7.62 , 7.54 , 3.37\u20133.35 , 1.85 . 13C NMR : 148.4, 133.5, 132.1, 131.5, 130.6, 123.9 (ArH), 48.2 (\u2013CH2NCH2\u2013), 25.9 (\u2013CH2CH2\u2013).2. o2.2.1.2. N-Piperidinyl-o-nitro\u00adbenzene\u00adsul\u00adfonamide, -Nitro\u00adbenzene\u00adsul\u00adfonyl chloride and piperidine . Yellow crystals ; RF\u00a0= 0.56 ; m.p. 91.6\u201391.8\u2005\u00b0C. IR : 3076 (aryl C\u2014H str.), 2947 (sp3-C\u2014H str.), 1552 (aryl C=C str.), 1550 (asym C\u2014NO2 str.), 1354 (sym C\u2014NO2 str.), 1350 (asym SO2\u2014N str.), 1166 (sym SO2\u2014N str.), 1056 (C\u2014N str.). 1H NMR : 7.96 , 7.70 , 7.59 , 3.26\u20133.24 , 1.64\u20131.63 , 1.55\u20131.54 . 13C NMR : 148.5, 133.6, 131.6, 131.5, 130.8, 123.5 (ArH), 47.0 (\u2013CH2NCH2\u2013), 25.4 (\u2013CH2CH2CH2\u2013), 23.5 (\u2013CH2CH2CH2\u2013).3. o2.2.1.3. N-Indolinyl-o-nitro\u00adbenzene\u00adsul\u00adfonamide, -Nitro\u00adbenzene\u00adsul\u00adfonyl chloride and indoline . Yellow crystals ; RF\u00a0= 0.79 ; m.p. 106.5\u2013106.8\u2005\u00b0C. IR : 3077 (aryl C\u2014H str.), 2976 (sp3-C\u2014H str.), 1594 (aryl C=C str.), 1536 (asym C\u2014NO2 str.), 1356 (sym C\u2014NO2 str.), 1355 (asym SO2\u2014N str.), 1161 (sym SO2\u2014N str.), 1051 (C\u2014N str.). 1H NMR : 7.95 , 7.71 , 7.62 , 7.48 , 7.21 , 7.05 , 4.17 , 3.10 . 13C NMR : 148.4, 141.1, 134.1, 131.8, 131.7, 131.6, 130.2, 127.8, 125.5, 124.3, 124.2, 114.5 (ArH), 50.5 (\u2013NCH2\u2013), 28.0 (\u2013NCH2CH2\u2013).N-cyclo\u00adamino-o-nitro\u00adbenzene\u00adsul\u00adfonamides 1\u20133 (15.63\u2005mmol) dissolved in ethanol (30\u2005ml), at ambient tem\u00adper\u00adature, and 10% palladium-on-charcoal catalyst (3.35\u2005mol%) was added, with stirring. Hydrogen gas was then introduced via a balloon and stirring continued at ambient tem\u00adper\u00adature for 12\u2005h. The reaction mixture was filtered and the solvent was evaporated in vacuo. The resulting residue was extracted into di\u00adchloro\u00admethane (50\u2005ml), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford an oil, which was purified on a silica-gel column using di\u00adchloro\u00admethane and n-hexane (2:1 v/v). Crystals were obtained via slow solvent evaporation of the eluates at ambient tem\u00adper\u00adature.An evacuated nitro\u00adgen-gas-filled round-bottomed flask was charged with 4. N2.2.2.1. N-Pyrrolidinyl-o-sul\u00adfan\u00adil\u00adamide, -Pyrrolidinyl-o-nitro\u00adbenzene\u00adsul\u00adfonamide 1 with 10% palladium-on-charcoal catalyst . Off-white crystals ; RF\u00a0= 0.40 ; m.p. 75.2\u201375.4\u2005\u00b0C. IR : 3464, 3363 (N\u2014H str.), 3003 (aryl C\u2014H str.), 2947 (sp3-C\u2014H str.), 1620 (aryl C=C str.), 1323 (asym SO2\u2014N str.), 1132 (sym SO2\u2014N str.), 1307 (C\u2014N str.). 1H NMR : 7.63 , 7.28 , 6.74 , 5.13 , 3.31 , 1.80 . 13C NMR : 146.4, 134.0, 130.2, 119.1, 117.6, 117.1 (ArH), 47.8 (\u2013CH2NCH2\u2013), 25.2 (\u2013CH2CH2\u2013).5. N2.2.2.2. N-Piperidinyl-o-sul\u00adfan\u00adil\u00adamide, -Piperidinyl-o-nitro\u00adbenzene\u00adsul\u00adfonamide 2 with 10% palladium-on-charcoal catalyst . Off-white crystals ; RF\u00a0= 0.57 ; m.p. 76.6\u201376.8\u2005\u00b0C. IR : 3487; 3383 (N\u2014H str.), 3072 (aryl C\u2014H str.), 2947 (sp3-C\u2014H str.), 1606 (aryl C=C str.), 1309 (asym SO2\u2014N str.), 1136 (sym SO2\u2014N str.), 1336 (C\u2014N str.). 1H NMR : 7.48 , 7.21 , 6.67 , 6.64 , 4.99 , 3.03\u20133.00 , 1.56\u20131.53 , 1.39\u20131.37 . 13C NMR : 146.3, 134.0, 130.1, 118.0, 117.6, 117.0 (ArH), 46.8 (\u2013CH2NCH2\u2013), 25.2 (\u2013CH2CH2CH2\u2013), 23.6 (\u2013CH2CH2CH2\u2013).6. N2.2.2.3. N-Indolinyl-o-sul\u00adfan\u00adil\u00adamide, -Indolinyl-o-nitro\u00adbenzene\u00adsul\u00adfonamide 3 with 10% palladium-on-charcoal catalyst . Off-white crystals ; RF\u00a0= 0.80 ; m.p.: 111.9\u2013112\u2005\u00b0C. IR : 3448, 3367 (N\u2014H str.), 3070 (aryl C\u2014H str.), 2924 (sp3-C\u2014H str.), 1597 (aryl C=C str.), 1327 (asym SO2\u2014N str.), 1138 (sym SO2\u2014N str.), 1330 (C\u2014N str.). 1H NMR : 7.48 , 7.16 , 7.08 , 7.03 , 6.90 , 6.58 , 6.55 , 5.00 , 3.96 , 2.86 . 13C NMR : 146.4, 142.3, 134.4, 132.1, 129.8, 127.6, 125.1, 123.7, 119.4, 117.7, 117.3, 115.1 (ArH), 50.0 (\u2013NCH2\u2013), 28.1 (\u2013NCH2CH2). Reaction synthesis of nitro\u00adsul\u00adfonamides 1\u20133 and amino\u00adsul\u00adfonamides 4\u20136 are presented in Scheme S1 in the supporting information. The FT\u2013IR, MS and 1H/13C NMR spectra of com\u00adpounds 1\u20136 are also presented in the supporting information.1\u20136 (CIF files) were imported directly into the Schr\u00f6dinger Suite -2-(thio\u00adphen-2-yl)acetamide (A) and 5-thio\u00adphene-2-sul\u00adfon\u00adamide (B) were based on chemical structures downloaded from the PubChem (https://pubchem.ncbi.nlm.nih.gov/) web\u00adsite in SDF format. A and B were used as reference com\u00adpounds because they are natural ligands in the crystalline state of 5fl4 and 4iwz. The Ligprep module of the mol\u00adecular model\u00adling platform of the Schr\u00f6dinger Suite Protein Data Bank (PDB). Retrieved crystal coordinates were prepared in the \u2018Protein Preparation Wizard\u2019 of the Schr\u00f6dinger Suite and XII (4iw7) isoforms will provide a selectivity profile that may be inter\u00adesting for the development of novel anti\u00adcancer agents with limited side effects. The hCA II (PDB entry 5fl4) and XII (4iw7) carbonic anhydrase iso\u00adforms have recently emerged as excellent targets for the design of novel therapeutic strategies for cancer, due to their involvement in the survival of tumour cells, as well as in the insurgence of resistance to classical anti\u00adcancer protocols mode of the 1\u20136 whereupon the SC-XRD structures of the com\u00adpounds were used for optimization and global reactivity descriptor (GRD) calculations. Computational studies and mol\u00adecular electrostatic potential (MEP) for 1\u20136 were carried out using the GAUSSIAN16 software package .Theoretical studies were performed for com\u00adpounds Crystal data, data collection and structure refinement details are summarized in Table\u00a01N-cyclo\u00adamino-o-sul\u00adfan\u00adil\u00adamides 4\u20136 were prepared via a two-step reaction, starting from the condensation reaction of o-nitro\u00adbenzene\u00adsul\u00adfonyl chloride with alicyclic amines in tolu\u00adene, at ambient tem\u00adper\u00adature, to afford N-cyclo\u00adamino-o-nitro\u00adbenzene\u00adsul\u00adfonamide adducts 1\u20133 (Scheme S1 in the supporting information). The use of toluene as a nonpolar reaction medium was, amongst other reasons, to drive the forward reaction. In the second step, adducts 1\u20133 were hydro\u00adgenated with hydro\u00adgen gas, in ethanol at ambient tem\u00adper\u00adature, in the presence of 10% palladium-on-activated charcoal catalyst to give the target N-cyclo\u00adamino-o-sul\u00adfan\u00adil\u00adamides 4\u20136 in 72\u201386% yield. The reactions were monitored by TLC.The 1H/13C NMR and MS spectra. In the IR spectra of o-nitro\u00adsul\u00adfonamide adducts 1\u20133, the strong absorption bands observed at 1355\u20131342 and 1171\u20131161\u2005cm\u22121 were ascribed to the asymmetric and symmetric stretching frequencies, respectively, of the SO2\u2014N moiety, thereby alluding to the formation of the sul\u00adfonamide bond. The disappearances of the SO2\u2014Cl (1420 and 1220\u2005cm\u22121) and N\u2014H (3286\u20133265\u2005cm\u22121) stretching bands in the IR spectra of o-nitro\u00adbenzene\u00adsul\u00adfonyl chloride and cyclo\u00adamines, respectively, were good indicators of a successful condensation reaction. This was corroborated by the shift of the sul\u00adfonyl (\u2013SO2\u2013) absorption bands from 1420 and 1220 (in o-nitro\u00adbenzene\u00adsul\u00adfonyl chloride) to 1355\u20131342 and 1171\u20131161\u2005cm\u22121 (in 1\u20133). It is noteworthy that the lower wavenumbers observed in the IR spectra of o-nitro\u00adsul\u00adfonamides 1\u20133 for \u2013SO2\u2013 were not unusual as the Cl atom bonded to it had been replaced by a less electronegative N atom. In the IR spectra of o-sul\u00adfan\u00adil\u00adamides 4\u20136, the appearance of two N\u2014H stretching bands in the higher frequency region around 3467\u2005\u00b1\u200520 and 3383\u2005\u00b1\u200510\u2005cm\u22121, and the disappearance of the nitro (NO2) absorption bands (observed at 1550\u20131536 and 1369\u20131342\u2005cm\u22121) in the spectra of 1\u20133 were attributed to the successful catalytic reduction of the nitro group to the amino group.All the com\u00adpounds synthesized were characterized by their melting points and IR, 1H NMR spectra of o-nitro\u00adsul\u00adfonamides 1\u20133 were additive of the individual spectra of the starting materials (i.e. o-nitro\u00adbenzene\u00adsul\u00adfonyl chloride and cyclo\u00adamines), with the disappearance of the nitro\u00adgen proton peaks of cyclo\u00adamines. The aromatic protons of o-sul\u00adfan\u00adil\u00adamides 4\u20136 resonated upfield in com\u00adparison to the same aromatic protons in precursors 1\u20133. This general shift towards tetra\u00admethyl\u00adsilane (TMS) was credited to the newly formed amino groups whose lone-pair electrons are suspected of having caused the increased mesomeric shielding of the aromatic protons. D2O-exchangeable singlets were also observed in the 1H NMR spectra of 4\u20136 between 5.13 and 4.99\u2005ppm for the newly-formed amino protons. The success of the catalytic hydro\u00adgenation of nitro adducts 1\u20133 was corroborated by the 13C NMR spectra of 4\u20136, where the requisite C atoms (C\u2014NO2 \u2192 C\u2014NH2) resonated upfield in the range 133.9\u2013130.1\u2005ppm. The spectroscopic data analyses of the synthesized com\u00adpounds were consistent with the assigned structures of the com\u00adpounds.The 1\u20133 and 4\u20136 crystallized in the monoclinic space group P21/n or P21/c (No. 14), except for 5, which crystallized in the ortho\u00adrhom\u00adbic space group Pbca (No. 61). In addition, they all had one mol\u00adecule in the asymmetric unit, with the exception of 4, with two independent mol\u00adecules per asymmetric unit cell. The two mol\u00adecules per unit cell of com\u00adpound 4 were identical but for the conformation of the pyrrolidine group . It is noteworthy that the pyrrolidine ring in 1 is disordered. The mol\u00adecular structures of 1\u20133 and 4\u20136 are shown in Fig.\u00a02o-nitro\u00adsul\u00adfonamides 1\u20133 and N-cyclo\u00adamino-o-sul\u00adfan\u00adil\u00adamides 4\u20136 are presented in Table\u00a01para-sul\u00adfan\u00adil\u00adamide and ortho-sul\u00adfan\u00adil\u00adamide, which crystallize in the ortho\u00adrhom\u00adbic Pbca (No. 61) and monoclinic P21/c (No. 14) space groups, respectively .The mol\u00adecules of 2) group in N-cyclo\u00adamino-o-sul\u00adfan\u00adil\u00adamides 4\u20136 contributed significantly to their hydro\u00adgen-bond inter\u00adactions (cf. Table\u00a02S(6) graph-set descriptors descriptors. Inter\u00adestingly, no infinite chain inter\u00adaction was observed in 4; instead, four mol\u00adecules were linked into a ring structure with an p-sul\u00adfan\u00adil\u00adamide strength \u2005\u00c5 were observed between the centrosymmetric indo\u00adline moieties. An N=O\u22ef\u03c0 ring inter\u00adaction of 3.657\u2005(2)\u2005\u00c5 was also evident in 3, whereas inter\u00admolecular C\u2014H\u22ef(\u03c0 ring) inter\u00adactions of 2.97\u2005\u00c5 and S=O\u22ef(\u03c0 ring) inter\u00adactions of 3.5773\u2005(15)\u2005\u00c5 were present in the structure of its hydro\u00adgenated analogue 6. The packing diagrams of the crystal structures of com\u00adpounds 1\u20136 are shown in Fig. S2 in the supporting information.The only \u03c0\u2013\u03c0 stacking inter\u00adaction of note occurred in et al., 20171\u20136 showed inter\u00admolecular inter\u00adactions such as O\u2014H\u22efO, O\u2014H\u22efN and C\u2014H\u22ef\u03c0. Two sharp O\u2014H spikes typical of an O\u2014H\u22efO inter\u00adaction from 1 contributed the highest O\u22efH inter\u00adaction of 42.3%. The fingerprint plots showed that C\u22efH contacts were highest for 6 (30.4%), and this is closely related to C\u2014H\u22ef\u03c0 inter\u00adactions visible on the dnorm surfaces typically indicate that the mol\u00adecule has a donor site(s) (e.g. amine and/or sul\u00adfone) or inter\u00adactions with proteins.The mol\u00adecular Hirshfeld surfaces, mapped as o-nitro\u00adsul\u00adfonamides 1\u20133 and N-cyclo\u00adamino-o-sul\u00adfan\u00adil\u00adamides 4\u20136 in full and resolved into C\u22efH, O\u22efH and N\u22efH are presented in Fig. S4 (supporting information). The inter\u00admolecular O\u22efH and N\u22efH inter\u00adactions appear as two distinct spikes of almost equal length in the 2D fingerprint plots in the region 1.2 < (de + di) < 2.9\u2005\u00c5 as light-sky-blue patterns in full fingerprint 2D plots and characterized to be 2.56\u2005\u00b1\u20050.21\u2005\u00c5 corresponds to O\u22efH contacts which contributes the majority of the surface area. 2D fingerprint plots reveal the contributions of these inter\u00adactions in the crystal structure qu\u00adanti\u00adtatively and are presented in Table\u00a04dnorm, di and de provided). Complementary regions are also visible in the fingerprint plots , where one mol\u00adecule acts as a donor (de > di) and the other acts as an acceptor (de < di). This finding was validated by the calculated mol\u00adecular electrostatic potential of 1\u20136 . The negative potential (acceptor) is indicated as a red surface around the O atoms attached to sulfur (\u2013SO2) and the N atoms attached to oxygen (\u2013NO2). The blue/purple surface area indicates that the positive potential (donor) is mapped in the proximity of the H atoms .Fingerprint plots of 1\u20136 presented bond lengths similar to those obtained from the crystal data. A com\u00adparison of selected torsion angles of the crystal structures of 1\u20136 and the DFT-optimized mol\u00adecules showed that con\u00adformation of the mol\u00adecules did not change significantly in the DFT-optimized state . Generally, the observed, almost flat, O\u2014S\u2014N\u2014C torsion angle of the DFT-optimized mol\u00adecules suggest that the lone pairs on sulfur may have con\u00adtributed to the \u03c0-electron delocalization that is observed in the DFT mol\u00adecules.The full geometry of optimized mol\u00adecules 3 and 5. The LUMO is mainly spread over the phenyl moieties. This indicates that there is a transfer of charge between the indolin\u00adyl/pyrrolidinyl rings and the phenyl moieties within the mol\u00adecule.The highest occupied mol\u00adecular orbital (HOMO) and lowest unoccupied mol\u00adecular orbital (LUMO) electrons are distributed around various moieties within the various mol\u00adecules Fig.\u00a04. Generalet al., 2020N-Indolinyl-o-nitro\u00adbenzene\u00adsul\u00adfonamide 3 displayed the smallest energy gap (3.24\u2005eV), indicating that it was the softest mol\u00adecule with good polarizability and reactivity, whereas N-piperidinyl-o-sul\u00adfan\u00adil\u00adamide 5 presented the largest energy gap of 4.924\u2005eV, thereby corroborating its high chemical hardness of 2.462\u2005eV (cf. Table\u00a053 (ELUMO\u00a0= \u22123.175\u2005eV), indicating that it is the best electron acceptor of the mol\u00adecules analyzed, whereas 6 was the best electron donor in the series, with the highest HOMO energy (EHOMO) of \u22126.142\u2005eV , electron affinity (A), chemical potential (\u03bc), electronegativity (\u03c7), global hardness (\u03b7), global softness (S) and global electrophilicity (\u03c9) values were calculated using the HOMO and LUMO energy values and are collated in Table\u00a05I value of 6.142\u2005eV originated from sul\u00adfan\u00adil\u00adamide 6, whereas sul\u00adfonamide 3 gave the largest A value of 3.175\u2005eV. Amongst the com\u00adpounds studied, 2 gave the highest \u03c7 value of 5.1795\u2005eV. Inter\u00adestingly, sul\u00adfan\u00adil\u00adamide 5 displayed the highest \u03b7 value of 2.462\u2005eV and the lowest chemical softness (S) of 0.406\u2005eV, thus alluding to its having the most reactive nature of all the mol\u00adecules investigated. The highest global electrophilicity of 29.597\u2005eV was also recorded for sul\u00adfonamide 2, indicating that it is a strong electro\u00adphile. In general, the chemical reactivities of com\u00adpounds 1\u20136 have been shown to vary with the groups attached to the com\u00adpounds -2-(thio\u00adphen-2-yl)acetamide (A) and 5-thio\u00adphene-2-sul\u00adfonamide (B) were also docked with respective proteins 4iwz and 5fl4, and taken as reference or standard drugs. Docking poses for the synthesized com\u00adpounds are displayed in Figs. S7\u2013S18, while those for the reference drugs are shown in Figs. 5Docking studies of synthesized 4iwz and A (reference drug) displayed a docking score of \u22122.252 kcal mol\u22121, which is higher than for all synthesized com\u00adpounds 1\u20136 and HIE64 (2.24\u2005\u00c5) via hydro\u00adgen-bonding inter\u00adactions and with amino acid residue HIS94 (4.75\u2005\u00c5) via \u03c0\u2013\u03c0 stacking inter\u00adactions and TRP5 (2.11\u2005\u00c5), and through \u03c0\u2013\u03c0 stacking with THR199 (1.81\u2005\u00c5) .Also, A Fig.\u00a05. Compoun5fl4, and the results obtained were com\u00adpared with the docked results of the reference drug B. We observed that the reference drug inter\u00adacts with amino acid residues ASP13 (1.59\u20132.73\u2005\u00c5) and VAL130 (2.53\u2005\u00c5) via hydro\u00adgen bonding, and with HID94 (5.49\u2005\u00c5) via \u03c0\u2013cation inter\u00adactions , displayed significantly good docking scores; however, they were lower than the reference drug .To determine the mode of inter\u00adaction of the synthesized com\u00adpounds with human carbonic anhydrase IX inhibitor (hCA IX), the synthesized com\u00adpounds were docked into the active site of 31 Fig.\u00a06. Benzene2 with 4iwz and 5fl4 are close to those obtained for A with 4iwz and B with 5fl4. Docking scores of mol\u00adecules with ring structures 1 and 3\u20136 also correlated with the electronegativity and electrophilicity values presented in Table\u00a05et al., 2018We observed that the docking scores of o-Nitro\u00adsul\u00adfonamides 1\u20133 and N-cyclo\u00adamino-o-sul\u00adfan\u00adil\u00adamides 4\u20136 have been successfully synthesized, characterized and the intermolecular interactions analysed, as well as being tested in silico for carbonic anhydrase II (4iwz) and IX (5fl4) inhibitory activities. The results obtained from crystal packing and DFT analysis suggests that the mol\u00adecules are held together by forces such as hydro\u00adgen bonding and \u03c0\u2013\u03c0 inter\u00adactions. The results of the DFT study of com\u00adpounds 1\u20136 were correlated with the mol\u00adecular docking data and indicate that electronegativity and electrophilicity of the title com\u00adpounds play an important role in their inter\u00adaction with carbonic anhydrase II (4iwz) and IX (5fl4).O-Nitro\u00adsul\u00adfonamide 2 displayed a good docking score against 4iwz (lower than the reference drug) and the best against 5fl4 (higher than the reference drug). These results provided a valuable synthesis approach and structural and docking information for com\u00adpounds 1\u20136 that may be used for the development of potent anti\u00adbacterial drugs.10.1107/S2053229622010130/oj3005sup1.cifCrystal structure: contains datablock(s) ka097, ja198, ja250, ja192, ka115, ja189, global. DOI: 10.1107/S2053229622010130/oj3005ka097sup2.hklStructure factors: contains datablock(s) ka097. DOI: 10.1107/S2053229622010130/oj3005ja198sup3.hklStructure factors: contains datablock(s) ja198. DOI: 10.1107/S2053229622010130/oj3005ja250sup4.hklStructure factors: contains datablock(s) ja250. DOI: 10.1107/S2053229622010130/oj3005ja192sup5.hklStructure factors: contains datablock(s) ja192. DOI: 10.1107/S2053229622010130/oj3005ka115sup6.hklStructure factors: contains datablock(s) ka115. DOI: 10.1107/S2053229622010130/oj3005ja189sup7.hklStructure factors: contains datablock(s) ja189. DOI: Click here for additional data file.10.1107/S2053229622010130/oj3005ka097sup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2053229622010130/oj3005ja198sup9.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2053229622010130/oj3005ja250sup10.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2053229622010130/oj3005ja192sup11.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2053229622010130/oj3005ka115sup12.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2053229622010130/oj3005ja189sup13.cmlSupporting information file. DOI: 10.1107/S2053229622010130/oj3005sup14.pdfAdditional figures, tables and spectra. DOI: 2014232, 2039638, 2014231, 2014230, 2014229, 2039639CCDC references:"} +{"text": "Fragment ligation with formylglycine occurred in\u2005situ in aqueous physiological buffer. Structures and kinetics were validated by NMR spectroscopy. Screening and hit validation revealed fluorinated and non\u2010fluorinated hit fragments being able to replace the native phosphotyrosine residue. The formylglycine probe identified low\u2010affinity fragments with high spatial resolution as substantiated by molecular modelling. The best fragment hit, 4\u2010amino\u2010phenyl\u2010acetic acid, was converted into a cellularly active, nanomolar inhibitor of the protein tyrosine phosphatase SHP2.Discovery of protein\u2010binding fragments for precisely defined binding sites is an unmet challenge to date. Herein, formylglycine is investigated as a molecular probe for the sensitive detection of fragments binding to a spatially defined protein site . Formylglycine peptide Site\u2010directed screening? Formylglycine (red) is a reactive amino acid that can be utilized in fragment ligation assays. Incorporated into a site\u2010directing hexapeptide it can effectively probe the active site of PTP1B, yielding new phosphotyrosine mimetics derived from different nucleophiles, such as aryl amines (green). DK values to enable detection.Site\u2010directed discovery of small protein\u2010binding fragments with M<250\u2005Da is a challenge due to the low affinities of most fragments and due to the lack of analytical methods that enable the detection of binding fragments at precisely defined positions on the protein surface.Fragment ligation has been introduced as a method to enhance binding of a primary fragment by a covalent, typically protein\u2010templated reaction with a secondary ligand.We reasoned that peptides and proteins carrying a reactive side chain electrophile should enable the site\u2010specific ligation of nucleophiles and thus might serve as tools for the site\u2010directed discovery of binding fragments for defined protein pockets Figure\u2005. FormylgThese findings encouraged us to investigate formylglycine peptides for the site\u2010directed discovery of protein\u2010binding fragments. As a model system, we selected the active sites of protein tyrosine phosphatases (PTP) which recognize and hydrolyze phosphotyrosine residues as substrates.1 (X=O) or the potent inhibitor 2 derived from it (X=CF2) and replaced the phosphotyrosine residue by formylglycine 1 contains the autophosphorylation sequence 988\u2013993 of epidermal growth factor receptor (EGF\u2212R) including the O\u2010phosphorylated tyrosine 992 and has been recognized as a substrate of the phosphotyrosine phosphatase PTP1B with a MK value of 3.6\u2005\u03bcM \u20104 was synthesized following a published 7\u2010step protocol from L\u2010serine,4 instead and established a facilitated access to 4 in only 3 steps starting from O\u2010ethyl 2\u2010nitro\u2010acetate 5. Treatment of 5 with titanium tetrachloride, N,N\u2010diisopropylethylamine, and triethyl orthoformate afforded the diethyl acetal 6 (conditions a).6 in the presence of Raney nickel furnished ethyl 2\u2010amino\u20103,3\u2010diethoxypropionate 7. Alternatively, under conditions b, N\u2010formylglycine ethylester 8 was formylated in C2 position with ethyl formate and tBuOK, followed by acetalization with HCl in EtOH at \u221225\u2009\u00b0C and cleavage of the N\u2010formyl group under basic conditions leading to the same diethylacetal 7.7 followed by N\u2010Fmoc protection with Fmoc\u2010OSu afforded the desired N\u2010Fmoc\u2010formylglycine 4 in a total yield of 68\u2009% over 4 steps or 43\u2009% over 3 steps .The phospho\u2010hexapeptide Ac\u2010Asp\u2010Ala\u2010Asp\u2010Glu\u2010Tyr*\u2010Leu\u2010NH4 was then used in solid phase peptide synthesis on polystyrene with Rink amide linker by diisopropyl carbodiimide/HOBt activation. Cleavage and complete deprotection was achieved with TFA/H2O. (95\u2009:\u20095 v/v) and the formylglycine peptide 3 was isolated by HPLC in 73\u2009% yield. Peptide 3 was highly soluble in water, buffer, and DMSO, so that we were able to analyze its structure and reactivity in solution .Building block 3 synthesized from (S)\u2010fGly and rac\u2010fGly were identical. In DMSO the fGly\u2010residue appears as a mixture of the aldehyde form 3, the hydrate form 9, and the enol form 10. In H2O/D2O 9\u2009:\u20091, exclusively the hydrate form 9 was observed with both diastereomeric H\u03b1 at 4.3\u2005ppm and both H\u03b2 at 5.3\u2005ppm. In the DQF\u2010H,H\u2010COSY spectrum all backbone NH protons were detectable suggesting that no cyclization with any of the backbone amides occurred and the formyl moiety thus was available for ligation. High resolution Q\u2010TOF MS analysis further confirmed the predominant hydration of the formylglycine peptide 3.NMR\u2010spectra of 3 were investigated with several nucleophilic fragments F detected in a primary screening experiment (see below) using NMR spectroscopy . One can suspect that the Z\u2010configuration of 11 is strongly favored over the E\u2010configuration due to an H\u2010bond between the enamine\u2010NH and the carbonyl residue of Glu4 resulting in a 7\u2010membered ring . Formation of the ligation product 11 was followed in Watergate 1H\u2005NMR for 1000\u2005min by integration of the benzylic CH2 group in starting material F1 and product 11 and indicated product formation with an average half reaction time of 10.7\u2005min \u2010trifluoromethyl sulfonamide F2 yielded enamine 12 formed hydrazone 13 with 100\u2009% conversion of 3 instantaneously (<2\u2005min). 13 was characterized by a shifted H\u03b1 of the fGly residue at 5.0 coupling with the H\u03b2 at 7.2 and the fGly\u2010NH at 8.3\u2005ppm. A single configuration of the hydrazone double bond was formed, presumably the thermodynamically favored E\u2010isomer. One might suspect as the preferred conformation the one establishing an H\u2010bonded, 6\u2010membered ring between the double\u2010bonded hydrazine\u2010N1 and the NH of Leu4 (1\u2009:\u20095), a common reducing agent in biochemistry and especially in assays with protein tyrosine phosphatases, yielded a ligation product detectable in HPLC\u2010MS with a mass corresponding to the hemi\u2010thioacetal 14 . 14 could not be detected in NMR or isolated, which is in full agreement with earlier results on the transient formation of bioactive hemi\u2010(thio)\u2010acetals formed by protein\u2010templated ligation reactions.3 DTT was found to compete with other nucleophiles such as amines or hydrazines. Therefore, it had to be replaced in enzyme assays with a non\u2010nucleophilic reducing agent. No ligation reaction was observed between formylglycine peptide 3 and tris\u2010carboxyethyl phosphane (TCEP) which was found to sustain the enzymatic activity of protein tyrosine phosphatases reliably at 50\u2005\u03bcM for several hours and therefore was employed in all ligation experiments.Incubation of 3 was tested as an inhibitor of two protein tyrosine phosphatases, PTP1B and SHP2, in an enzyme activity assay using DiFMUP as a fluorogenic substrate. Peptide 3 bound to and inhibited PTP1B with a IK value of 484\u2005\u03bcM and SHP2 with a IK of 341\u2005\u03bcM were selected and tested as inhibitors of PTP1B alone and in combination with peptide 3 . Fragments were pre\u2010selected for library composition based upon representation of potential phosphate\u2010mimetic substructures including carboxylic acids, sulfonic acids, sulfonamides, and fluorine\u2010rich functional groups, which were to be investigated for fluorine\u2010specific interactions with phosphotyrosine binding sites., 687.2711\u2005Da. calcd m/z 688.2790 [M+H]+, 710.2609 [M+Na]+, found m/z 688.2792 [M+H]+, 710.2607 [M+Na]+.3,3\u2010Diethoxy\u20102\u2010(9H\u2010fluoren\u20109\u2010ylmethoxycarbonylamino)\u2010 propanoic acid 4: Ethyl 2\u2010amino\u20103,3\u2010diethoxypropanoate 7 was suspended in 0.5\u2005M aq. LiOH solution (5\u2005mL) and methanol was added until a clear solution remained. It was stirred at room temperature for 2\u2005h. TLC control after this time revealed the hydrolysis of 7 after which the mixture was neutralized with Amberlite\u00ae IR\u2010120 loaded with H+. The resin was filtered off and washed with water and concentrated under reduced pressure. The product of hydrolysis and NaHCO3 were stirred in H2O (10\u2005mL) at 0\u2009\u00b0C. To this solution was added dropwise 9\u2010fluorenylmethyl N\u2010succinimidyl carbonate dissolved in 1,4\u2010dioxane (10\u2005mL) at 0\u2009\u00b0C. The mixture was stirred for 1\u2005h at 0\u2009\u00b0C, warmed to room temperature and then stirred for another 12\u2005h. The reaction mixture was concentrated under reduced pressure and purified by column chromatography to provide 4 as white solid ; Rf=0,46 . 1H NMR: : \u03b4=7.75 , 7.61 , 7.39 , 7.31 , 5.67 , 4.85 , 4.59 , 4.39 , 4.24 , 3.88\u20133.77 , 3.76\u20133.6 , 3.65\u20133.57 , 1.32 , 1.21 ppm. 13C NMR: : \u03b4=172.33 (COOH), 156.59 (CONH), 143.80 (Ar\u2212Cquart), 141.37 (Ar\u2212Cquart), 127.83 (Ar\u2212C), 127.19 (Ar\u2212C), 125.28 (Ar\u2212C), 120.06 (Ar\u2212C), 101.17 (C\u03b2), 67.53 (Fmoc CH2), 64.74 (CH2CH3), 64.69 (CH2CH3), 56.73 (C\u03b1), 47.15 (Fmoc CH), 15.20 (CH2CH3), 15.08 (CH2CH3) ppm. HRMS: (ESI): C22H25NO6 [M], 399.1682\u2005Da. calcd m/z 398.1604 [M\u2212H]\u2212, 422.158 [M+Na]+, found m/z 398.1609 [M+H]+, 422.1578 [M+Na]+.Ethyl 3,3\u2010diethoxy\u20102\u2010nitropropanoate 6: To a solution of ethyl\u2010nitroacetate 5 in anhydrous methylene dichloride (25\u2005mL) held at \u2010 10\u2009\u00b0C under an argon atmosphere was slowly added by syringe titanium(IV)\u2010chloride . The mixture was stirred for 10\u2005min and N,N\u2010diisopropylethylamine was added to the mixture dropwise over 30\u2005min. The Mixture was held at \u221210\u2009\u00b0C with stirring for 1\u2005h. triethyl\u2010orthoformate was added to the mixture dropwise and stirring was continued for 2\u2005h at \u221210\u2009\u00b0C. The reaction mixture was diluted with a 20\u2009% solution of ethanol in saturated aqueous NaHCO3 (100\u2005mL) and the mixture was stirred vigorously for 10\u2005min. Organic solvents were removed from the mixture under reduced pressure. Water (200\u2005mL) was added to the concentrated reaction mixture and the aqueous phase was extracted three times with ethylacetate. The combined organic phases were filtered over celite and dried over Na2SO4. Evaporation of the solvents under reduced pressure afforded compound 6 as yellow oil ; Rf=0,46 . 1H NMR: : \u03b4=5.24 , 5.17 , 4.28 , 3.81\u20133.75 , 3.69\u20133.61 , 1.30 , 1.19 ppm. 13C NMR: : \u03b4=161.77 (COOEt), 100.08 (C\u03b2), 88.52 (C\u03b1), 64.79 (OCH2CH3), 64.60 (OCH2CH3), 63.18 (COOCH2CH3), 15.16 (OCH2CH3), 15.07 (OCH2CH3), 13.94 (COOCH2CH3) ppm. HRMS: C9H17NO6 [M], 235.1056\u2005Da. calcd m/z 258.0948 [M+Na]+, 274.0687 [M+K]+, found m/z 258.0948 [M+Na]+, 274.0680 [M+K]+.Ethyl 2\u2010amino\u20103,3\u2010diethoxypropanoate 7 (Method A): Ethyl 3,3\u2010diethoxy\u20102\u2010nitropropanoate 6 was dissolved in absolute ethanol (5\u2005mL) and hydrogen was bubbled through the mixture for 5\u2005min. raney nickel was added and the mixture was hydrogenated at atmospheric pressure for 12\u2005h at room temperature. The reaction mixture was filtered through celite, and the filter cake was washed plentiful with ethanol. Filtrate was concentrated under reduced pressure, leaving a reddish oil. Vacuum distillation of this afforded 7 as yellowish oil ; Rf=0,29 . 1H NMR: : \u03b4=4.61 , 4.24\u20134.16 , 3.77\u20133.67 , 3.61 , 3.58\u20133.51 , 2.05\u20131.95 1.30\u20131.25 , 1.23\u20131.19 , 1.19\u20131.15 ppm. 13C NMR: : \u03b4=161.8 (COOEt), 100.1 (C\u03b2), 88.5 (C\u03b1), 64.8 (OC2CH3), 64.60 (OC2CH3), 63.2 (COOC2CH3), 15.2 (OC2CH3), 15.1 (OC2CH3), 13.9 (COOC2CH3) ppm. HRMS: (ESI): C9H19NO4 [M], 205.1314\u2005Da. calcd m/z 206.1392 [M+H]+, 228.1206 [M+Na]+, found m/z 206.1379 [M+H]+, 228.1195 [M+Na]+.Ethyl 2\u2010amino\u20103,3\u2010diethoxypropanoate 7 (Method B): A solution of N\u2010formylglycine ethylester 8 in ethylformate was added dropwise to a mixture of potassium tert\u2010butoxide in toluene (60\u2005mL) at 10\u2009\u00b0C over a periode of 2\u2005h. Stirring was continued for additional 2\u2005h after which the mixture was allowed to stand at 4\u2009\u00b0C for 18\u2005h. The supernatant was discarded and the gelantinious residue dissolved in ethanol (35\u2005mL). This solution was the diluted with methylene dichloride (50\u2005mL) and cooled to \u221225\u2009\u00b0C and treated with HCl gas for 3\u2005h. The mixture was then stirred for 24\u2005h at room temperature. The solution was concentrated under reduced pressure and the residue was suspended in diethyl ether (80\u2005mL). This suspension was treated with saturated K2CO3 until strongly basic when the phases were separated, and the organic phase was washed further with water and dried over Na2SO4 and evaporated under reduced pressure to afford an oil. Vacuum distillation of this afforded 8 as yellowish oil .NMR ligation experiments: The ligation of peptide 3 with fragments F1, F2 or F3 was confirmed by NMR experiments. The respective fragments were dissolved in a solution of 3, to yield a sample with a final concentration of 10\u2005mM of the fragment and 5\u2005mM of 3. Experiments were performed in 9\u2009:\u20091 H2O/D2O or buffer at 300\u2005K. Experiments were performed using a WATERGATE water suppression.Peptide 15: Formylglycine peptide 3 and amine F1 were stirred in dry MeOH (1\u2005mL) and AcOH (20\u2005\u03bcL) with molecular sieve for 1\u2005h at room temperature. NaCNBH3 (2.5\u2005equiv) was added and the mixtures was stirred for 18\u2005h. Molecular sieves were filtered off and the mixture was concentrated under reduced pressure. The residue was purified by RP flash column chromatography yielding 15 as a white solid ; HRMS: (ESI): C35H50N8O15 [M], 822.3396\u2005Da. calcd m/z 823.3474 [M+H]+, 821.3317 [M\u2212H]\u2212, found m/z 823.3477 [M+H]+, 821.3316 [M\u2212H]\u2212.Peptide 16: Following the procedure for peptide 15, 3 and F4 yielded 16 as white solid ; HRMS: (ESI): C35H50N8O16 [M], 838.3345\u2005Da. calcd m/z 839.3423 [M+H]+, 837.3267 [M\u2212H]\u2212, found m/z [M+H]+, [M\u2212H]\u2212.Peptide 17: Following the procedure for peptide 15, 3 and barbituric acid yielded 17 as white solid ; HRMS: (ESI): C31H45N9O16 [M], 799.2984\u2005Da. calcd m/z 800.3063 [M+H]+, 798.2906 [M\u2212H]\u2212, found m/z 800.3058 [M+H]+, 798.2912 [M\u2212H]\u2212.Peptide 18: Following the procedure for peptide 3, using the unnatural amino acid 19, 18 was obtained as a white solid from 200\u2005mg resin. 1H\u2005NMR: : \u03b4=8.21 , 8.16 , 8.06 , 8.03 , 7.97 , 7.78 , 7.23 , 7.04 , 6.98 , 6.64 , 4.59\u20134.54 , 4.54\u20134.50 , 4.44 , 4.30\u20134.24 , 4.26\u20134.24 , 4.21 , 4.18 , 3.34 , 3.23 , 2.77\u20132.69 , 2.69\u20132.66 , 2.59\u20132.55 , 2.26\u20132.19 , 1.97\u20131.89 , 1.85 , 1.81\u20131.73 , 1.63\u20131.56 , 1.51\u20131.45 , 1.22 , 0.87 , 0.82 . HRMS: (ESI): C34H48F3N9O15S [M], 911.2943\u2005Da. calcd m/z 912.3021 [M+H]+, 934.2840 [M+Na]+, found m/z 912.3021 [M+H]+, 934.2838 [M+Na]+.2\u2010N\u2010(9H\u2010Fluoren\u20109\u2010ylmethoxycarbonylamino)\u20103\u2010(4\u2010trifluoromethyl\u2010sulfonamido)\u2010phenyl)\u2010amino\u2010propanoic acid 19: To a solution of 20 and NaHCO3 in water (10\u2005mL) was added N\u2010(9\u2010fluorenylmethoxy\u2010carbonyloxy)\u2010succinimide dissolved in 1,4\u2010dioxane (10\u2005mL) dropwise at 0\u2009\u00b0C. The mixture was allowed to slowly warm up to room temperature and was subsequently stirred for 18\u2005h. Water was added (25\u2005mL) and the mixture extracted with ethyl acetate (3\u00d750\u2005mL). The organic layer was washed with 0.2\u2005M HCl (40\u2005mL) and brine (40\u2005mL) and dried over Na2SO3. The solvents were removed under reduced pressure and the crude product purified via MPLC yielding 19 as a brown solid . 1H NMR: \u03b4=7.90 , 7.72 , 7.68 , 7.42 , 7.33 , 7.31 , 6.99 , 6.63 , 4.35\u20134.29 , 4.24 , 4.23\u20134.20 , 3.42 , 3.36 . 13C NMR: \u03b4=172.88 (COOH), 156.62 (CONH), 147.93 (Ar\u2212Cquart), 144.25 (Fmoc Ar\u2212Cquart), 141.18 (Fmoc Ar\u2212Cquart), 128.09 (Fmoc Ar\u2212C), 127.54 (Fmoc Ar\u2212C), 127.17 (Fmoc Ar\u2212C), 125.68 (Fmoc Ar\u2212C), 122.78 (Ar\u2212Cquart), 121.26, 120.57 (Ar\u2212C), 119.42 (CF3), 112.83 (Ar\u2212C), 66.17 (Fmoc CH), 53.75 (C\u03b1), 47.08 (Fmoc CH2), 44.52 (C\u03b2). HRMS: (ESI): C25H22F3N3O6S [M], 549.1181\u2005Da. calcd m/z 550.1260 [M+H]+, 572.1079 [M+Na]+, found m/z 550.1260 [M+H]+, 572.1080 [M+Na]+.2\u2010Amino\u20103\u2010(4\u2010trifluoromethyl\u2010sulfonamido\u2010phenyl)\u2010amino\u2010propanoic acid 20: A mixture of L\u20102\u2010N\u2010Boc\u20102,3\u2010diamino\u2010propanoic acid (Boc\u2010Dap\u2010OH) , 1\u2010fluoro\u20104\u2010nitrobenzene and K2CO3 in ethanol (15\u2005mL) was stirred for 24\u2005h at 90\u2009\u00b0C. To the crude residue of this reaction was added 10\u2009% Pd/C and the flask wash flushed with H2 and left under H2 atmosphere for 18\u2005h at room temperature. The catalyst was filtered off, the solvent evaporated under vacuum and purified by column chromatography to provide N\u20102\u2010Boc\u20103\u2010(4\u2010aminophenyl)\u20102,3\u2010diamino\u2010propanoic acid as brownish solid . 1H NMR: \u03b4=6.41 , 6.33 , 6.08 3.64 , 3.11 , 2.94 , 1.38 . 13C NMR: \u03b4=172.95 (COOH), 155.55 (Boc CO), 141.09 (Ar\u2212Cquart), 139.52 (Ar\u2212Cquart), 116.07 (Ar\u2212C), 114.29 (Ar\u2212C), 78.08 (Boc Cquart), 54.97 (C\u03b1), 48.36 (C\u03b2), 28.79 (Boc CH3). HRMS: (ESI): C14H21N3O4 [M], 295.1532\u2005Da. calcd m/z 296.1610 [M+H]+, 294.1454 [M\u2212H]\u2212, found m/z 296.1612 [M+H]+, 294.1451 [M\u2212H]\u2212. The obtained intermediate was dissolved in anhydrous DCM (10\u2005mL) under an argon atmosphere and the reaction mixture was cooled to 0\u2009\u00b0C. Trifluoromethanesulfonic anhydride was added dropwise to the reaction mixture and stirred for 30\u2005min at 0\u2009\u00b0C, after which the mixture was slowly warmed to room temperature and stirred further for 18\u2005h. The reaction was diluted with water (20\u2005mL), filtered and the feed was washed twice with MeCN/H2O . The solvents were evaporated under vacuum and purified by column chromatography to provide 20 as brownish solid . 1H NMR: \u03b4=7.20 , 6.86 , 6.77 , 6.45 , 3.53 , 3.42 , 3.22 . 13C NMR: \u03b4=174.37 (C\u20102), 143.13 (C\u20105), 125.16 (C\u20106), 123.58 (C\u20108), 118.43 (C\u201010), 113.56 (C\u20107), 53.77 (C\u20101), 45.69 (C\u20103). HRMS: (ESI): C10H12F3N3O4S [M], 327.0501\u2005Da. calcd m/z 328.0579 [M+H]+, 326.0422 [M\u2212H]\u2212, found m/z 328.0582 [M+H]+, 326.0419 [M\u2212H]\u2212.Z)\u20102\u2010\u20105\u2010oxo\u20101,5\u2010dihydro\u20104H\u2010pyrazol\u20104\u2010ylidene)\u2010hydrazinyl)\u2010phenyl)\u2010acetic acid (21)(: A solution of NaNO2 (0.17\u2005mmol) in H2O (100\u2005\u03bcL) was added at 0\u2009\u00b0C to a suspension of 2\u2010(4\u2010aminophenyl)\u2010acetic acid (0.17\u2005mmol) in 4\u2005M HCl (0.3\u2005mL). The acidic solution was stirred for 1\u2005h at 0\u2009\u00b0C, after which a solution of 2,5\u2010bis(4\u2010nitrophenyl)\u20102,4\u2010dihydro\u20103H\u2010pyrazol\u20103\u2010one (0.15\u2005mmol) in THF (2\u2005mL) was added and the mixtures was stirred for 10\u2005min at 0\u2009\u00b0C. Afterwards a solution of 1.3\u2005M NH4OH (0.2\u2005mL) was added, stirred for 1\u2005h at 0\u2009\u00b0C and for 24\u2005h at r.t. The suspension was filtered under suction, the solid was washed with H2O (2\u00d720\u2005mL), and dried in air. A mixture of EtOAc:DCM:n\u2010Hex was added in a sealed flask for 2\u2005h. The suspension was filtered off, washed with Et2O and dried under vacuo to provide product 21. 1H\u2005NMR: : \u03b4=8.55 , 8.51 , 8.34 , 8.29 , 7.54 , 7.29 , 3.56 ppm. 13C\u2005NMR: : \u03b4=173.6 (COOH) 157.9 (C\u20107), 147.1 (Pyr Cquart), 146.3 (Ar\u2212Cquart), 143.88 (Pyr Cquart), 142.4 (Ar\u2212Cquart), 141.3 (Ar\u2212Cquart), 130.4 (Ar\u2212C), 130.0 (Ar\u2212Cquart), 129.1(Ar\u2212C), 125.4 (Ar\u2212C), 123.9 (Ar\u2212C), 122.3 (Ar\u2212Cquart), 121.2 (Ar\u2212Cquart), 120.9 (Ar\u2212C), 117.6 (Ar\u2212C), 32.0 (CH2) ppm. HRMS: (ESI): C23H16N6O7 [M], 488.1080\u2005Da. calcd m/z 489.1159 [M+H]+, 487.1002 [M\u2212H]\u2212, found m/z 489.1162 [M+H]+, 487.1005 [M\u2212H]\u2212. Anal: calcd for C22H14F3N7O7S: C, 56.56; H, 3.30; N, 17.21; found: C, 57.12; H, 3.58; N, 17.56.(Z)\u20104\u2010\u20105\u2010oxo\u20101,5\u2010dihydro\u20104H\u2010pyrazol\u20104\u2010ylidene)\u2010hydrazinyl)\u2010phenyl)\u20101,1,1\u2010trifluoromethanesulfonamide (22): Following the procedure for compound 21, using N\u2010(4\u2010aminophenyl)\u20101,1,1\u2010trifluoromethanesulfonamide and 2,5\u2010bis(4\u2010nitrophenyl)\u20102,4\u2010dihydro\u20103H\u2010pyrazol\u20103\u2010one , 22 was obtained as an orange solid . HRMS: (ESI): C22H14F3N7O7S [M], 577.0628\u2005Da. calcd m/z 578.0706 [M+H]+, found m/z 578.0696 [M+H]+. Anal: calcd for C22H14F3N7O7S: C, 45.76; H, 2.44; N, 16.98; found: C, 45.78; H, 2.54; N, 16.98.Z)\u20102\u2010\u20105\u2010oxo\u20101,5\u2010dihydro\u20104H\u2010pyrazol\u20104\u2010ylidene)\u2010hydrazinyl)\u2010phenoxy)\u2010acetic acid (23)\u2010acetic acid and 2,5\u2010bis(4\u2010nitrophenyl)\u20102,4\u2010dihydro\u20103H\u2010pyrazol\u20103\u2010one , 23 was obtained as an orange solid . HRMS: (ESI): C23H16N6O8 [M], 504.1030\u2005Da. calcd m/z 505.1108 [M+H]+, 503.0951 [M\u2212H]\u2212, found m/z 505.1106 [M+H]+, 503.0976 [M\u2212H]\u2212. Anal: calcd for C23H16N6O8: C, 54.77; H, 3.20; N, 16.66; found: C, 55.23; H, 3.18; N, 15.62.Protein Expression and purification: DNA encoding the catalytic domain of human PTP1B (amino acids 1\u2013321) was subcloned into the pQLinkH vector. The gene encoding the N\u2010terminal His7\u2010tagged protein was over\u2010expressed at 17\u2009\u00b0C in E. coli Rosetta (DE3). The purification procedure comprises an affinity chromatography on a 5\u2005mL HisTrap FF crude column , charged with Ni2+, and a size\u2010exclusion chromatography on a Superdex 26/60 column equilibrated with 25\u2005mM HEPES\u2010NaOH (pH=7.5), 50\u2005mM NaCl, 1\u2005mM DTT. The His7\u2010tag was cleaved with tobacco etch virus protease at 20\u2009\u00b0C prior to the gel\u2010filtration step. The catalytic domain of human SHP2 (amino acids 225\u2013541) was purified from E. coli B21 (DE3). The overexpressed protein was fused to an N\u2010terminal His10\u2010tag. Cells were harvested and resuspended in 25\u2005mM Tris\u2010HCl (pH=7.5), 50\u2005mM NaCl, 1\u2009% (v/v) Triton X\u2010100, 10\u2009% (v/v) glycerol, 1\u2005mM DTT. After cell lysis, debris was removed by centrifugation at 12,000\u2005rpm for 30\u2005min. The supernatant was filtered prior to affinity purification with Ni\u2010NTA agarose (Qiagen). The slurry was washed with 50\u2005mM Tris\u2010HCl (pH=8.0), 500\u2005mM NaCl supplemented with 25\u2005mM imidazole. Reasonably pure protein was eluted with 200\u2013300\u2005mM imidazole. The purification was completed by size\u2010exclusion chromatography with a Superdex 75 16/600 column equilibrated with 20\u2005mM HEPES\u2010NaOH (pH=7.5), 50\u2005mM NaCl, 1\u2005mM DTT.Enzyme activity assays of SHP2 and PTP1B: The catalytic activity of SHP2 catalytic domain and PTP1B were monitored using the fluorogenic substrate DiFMUP . Phosphatase reactions were performed at room temperature in 384\u2010well black plate, clear flat bottom, low flange, non\u2010binding surface using a final volume of 20\u2005\u03bcL and the following assay buffer conditions: 50\u2005mM MOPS, pH=6.5, 200\u2005mM NaCl, 50\u2005\u03bcM TCEP, 0.03\u2009% Tween\u201020 (freshly added prior to each measurement). Test compounds were dissolved in DMSO or buffer at stock concentrations of 50, 20 or 10\u2005mM and serially diluted. Enzyme and different concentrations of the tested compounds were incubated in buffer for 30\u201360\u2005min at room temperature. Measurements were performed with a final concentration of 2.5\u2009% DMSO unless stated otherwise at 37\u2009\u00b0C and were performed in triplicate. Enzymatic reactions were started by adding DiFMUP concentrations matching the experimentally determined MK values of the enzymes of 67\u2005\u03bcM (PTP1B) or 72\u2005\u03bcM (SHP2). Samples were excited at a wavelength of 360\u2005nm and emitted fluorescence was recorded at 460 for 10\u2005min using a microplate reader . Initial slope of fluorescence was determined in triplicate and 50IC values were calculated with GraphPad Prism 5. Determined 50IC values were converted into the corresponding IK values applying the Cheng Prusoff equation IK=50IC/(1+[S]/MK ).Dynamic ligation screening of nucleophilic fragments with 3: Under assay conditions described above, PTP1B or SHP2 were pre\u2010incubated with 3 (100\u2005\u03bcM) for 5\u2005min. Subsequently, (200\u2005\u03bcM) of the respective fragments were added and the resulting mixtures were incubated for 30\u2005min at RT, prior to DiFMUP addition.Determination of apparentKI\u2010values of 11\u2009in a time\u2010dependent assay: Under the conditions described above, PTP1B was pre\u2010incubated with peptide 3 in different concentrations for 5\u2005mins. Subsequently, over the course of 60\u2005mins (every 15\u2005min), 2 equivalents of fragment F1 were added to each of the different concentrations of 3 at RT, prior to DiFMUP addition.Determination of apparentKI\u2010values of 12\u2009in a concentration\u2010dependent assay: Under the conditions described above, PTP1B was pre\u2010incubated with peptide 3 at different concentrations for 5\u2005min. Subsequently, fragment F2 was added at three different concentrations and the resulting mixtures were incubated for 30\u2005min at RT, prior to DiFMUP addition.Computational Methods: The protein X\u2010ray diffraction crystal structure of mutated PTP1B (PDB code: 1PTU)3, 9, 11, 12, 15, 16, and 18 were docked to the binding pocket of PTP1B using Schr\u00f6dinger's GLIDE.Cellular experiments: HeLa cells were cultivated in DMEM buffer with 10\u2009% FCS in 75\u2005cm2 cell culture flasks at 37\u2009\u00b0C, 5\u2009% CO2. At confluency of about 75\u2009%, cells were seeded in a density of 250,000\u2005cells/mL in 6\u2010well plates and incubated for 24\u2005h. Subsequently, medium was removed and replaced by DMEM buffer with 0.1\u2009% BSA (1\u2005mL per well) and incubated for another 16\u2005h. Hepatocyte growth factor and test compounds in different final concentrations were added. Control wells received the same amount of DMSO or no addition. Plates were incubated for 1\u2005h, then washed with cold PBS and shaken with lysis buffer for 5\u2005min. Cell lysates were transferred to Eppendorf cups and centrifuged at 4\u2009\u00b0C with 10,000\u2005g for 10\u2005min. Total protein was quantified in cell lysates using RotiNanoquant (Carl Roth # K880.1) and 30\u2005\u03bcg of protein was applied to 12\u2009%\u2010SDS\u2010PAGE with a run time of 90\u2005min at 150\u2005V. Then protein was blotted in Towbin bufer to an PVDF\u2010membrane over 1\u2005h at 100\u2005V, and the membrane was saturated with 20\u2005ml TBS\u2010Tween/ 2\u2009% BSA, 1\u2005h at RT. The blot was incubated with anti\u2010phospo\u2010ERK as primary antibody overnight at 4\u2009\u00b0C, washed 3\u00d75\u2005min with TBS\u2010Tween, and incubated with anti\u2010mouse IgG\u2010hrp as secondary antibody . The blot was again washed 3\u00d75\u2005min with TBS\u2010Tween and imaged with the ECL system (ThermoScientific # 34080) at Syngene PXi Imager. From the same lysis sample, under the same conditions, ERK 1/2 and \u03b2\u2010tubulin were blotted. For \u03b2\u2010tubulin, the antibody incubation time was adjusted to 2\u2005h at RT.The authors declare no conflict of interest.1As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "The chains are inter\u00addigitated.The nearly planar mol\u00adecule is centrosymmetric with two all- 70H112N6O6, is centrosymmetric with two all-s-trans chains, the other two chains have an s-cis unit starting with the oxygen atoms. The chains are inter\u00addigitated in the packing.The nearly planar mol\u00adecule of the title compound, C The centrosymmetric mol\u00adecule is located parallel to the (110) plane, the five aromatic rings are almost coplanar with torsion angles of \u22120.4\u2005(4)\u00b0 (N6\u2014C5\u2014C3\u2014C2) and \u22122.0\u2005(4)\u00b0 (C11\u2014C10\u2014C8\u2014N7). Two dodec\u00adyloxy chains per mol\u00adecule are completely all-s-trans organized whereas the other pair shows an s-cis-conformation of the O16\u2014C17\u2014C18\u2014C19 unit. Torsion angles at the all-trans chains are 179.3\u2005(2) for C13\u2014C14\u2014O29\u2014C30, 177.9\u2005(5)\u00b0 for C14\u2014O29\u2014C30\u2014C31 and 178.1\u2005(2)\u00b0 for O29\u2014C30\u2014C31\u2014C32 but for the other chain, 15.8\u2005(3)\u00b0 (C17\u2014O16\u2014C12\u2014C13), 176.2\u2005(2)\u00b0 (C12\u2014O16\u2014C17\u2014C18), and \u221255.3\u2005(3)\u00b0 for the O16\u2014C17\u2014C18\u2014C19 unit. The packing is controlled by inter\u00adaction of the aliphatic chains is located above the pyrazine centroid.Electron-deficient conjugated oligomers are inter\u00adesting as electron-transporting mat\u00aderials in organic electronics : 2916, 2848, 1602, 1544, 1468, 1442, 1429, 1174; 1H NMR , 6.66 , 4.04 , 3.17 , 1.82 , 1.55-1.42 , 1.42-1.20 , 0.93-0.83 . Whereas the absorption spectrum of the title compound in cyclo\u00adhexane peaks at 348\u2005nm, three maxima appear in methanol , indicating aggregation. The emission of a solution in cyclo\u00adhexane is centered at 403\u2005nm, and in toluene at 409\u2005nm. Crystallization was via slow evaporation of a solution in chloro\u00adform/2-propanol.The title compound was prepared Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2414314623000342/bt4131sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2414314623000342/bt4131Isup2.hklStructure factors: contains datablock(s) I. DOI: 2235881CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit contains three independent mol\u00adecules differing slightly in conformation. Portions of the observed conformations are determined by intra\u00admolecular N\u2014H\u22efO hydrogen bonds. In the crystal, O\u2014H\u22efO hydrogen bonds form chains of mol\u00adecules which are linked into corrugated sheets parallel to ( 14H13NO4, contains three independent mol\u00adecules, which differ slightly in conformation. Each contains an intra\u00admolecular N\u2014H\u22efO hydrogen bond. In the crystal, O\u2014H\u22efO hydrogen bonds form chains of mol\u00adecules, which are linked into corrugated sheets parallel to (The asymmetric unit of the title compound, C These compare quite favorably with those found in mol\u00adecules with R = Me differ modestly in the orientations of the methyl groups while the third differs more in conformation from the other two Fig.\u00a01. In each3.B\u22efO7, O5\u2014H5B\u22efO11 and O9\u2014H9B\u22efO3 hydrogen bonds repeating in that order \u2005\u00c5, C10\u22efCg2 = 3.731\u2005(2)\u2005\u00c5, C10=O2\u22efCg2 = 91.41\u2005(13)\u00b0 \u2005\u00c5, C24\u22efCg6 = 3.694\u2005(2)\u2005\u00c5, C24=O6\u22efCg6 = 91.12\u2005(14)\u00b0 ; O10\u22efCg4 = 3.4110\u2005(18)\u2005\u00c5, C38\u22efCg4 = 3.656\u2005(2)\u2005\u00c5, C38=O10\u22efCg4 = 91.00\u2005(13)\u00b0 ]. The layers are held together by C11\u2014H11C\u22efO6 hydrogen bonds are smaller than 1.0% with low densities of points.In order to visualize the inter\u00admolecular inter\u00adactions, a Hirshfeld surface (HS) analysis using the standard B3LYP functional and 6\u2013311\u2005G basis-set calculations , hardness (\u03b7), potential (\u03bc), electrophilicity (\u03c9) and softness (\u03c3) are recorded in Table\u00a03E)-3-[1-(2-hy\u00addroxy\u00adphenyl\u00adamino)\u00adethyl\u00adidene]-6-methyl-3H-pyran-2,4-dione ring. The energy band gap [\u0394E = ELUMO\u00a0\u2212\u00a0EHOMO] of the mol\u00adecule is 4.54\u2005eV, and the frontier mol\u00adecular orbital energies, EHOMO and ELUMO are \u22126.12 and \u22121.58\u2005eV, respectively.The optimized structure of the title compound in the gas phase was generated theoretically 6.Gaussview software was used to broadly predict reactive sites for electrophilic and nucleophilic attack in the title compound by B3LYP/6-31G optimized geometries using 7.et al., 2016A yielded 66 hits of which 15 were deemed most similar to the title mol\u00adecule. These include mol\u00adecules with R = Me in 30\u2005mL of ethanol, 2.5\u2005mmol of de\u00adhydro\u00adacetic acid were added. The mixture was refluxed for 1\u2005h. After cooling, the precipitate that formed was recrystallized from ethanol solution to give yellow crystals in 88% yield.9.Uiso(H) = 1.2Ueq or 1.5Ueq.Crystal, data collection and refinement details are presented in Table\u00a0410.1107/S2056989022007514/zn2020sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989022007514/zn2020Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022007514/zn2020Isup3.cmlSupporting information file. DOI: 2192044CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N\u2010Acetylneuraminic acid is one of a large, diverse family of nine\u2010carbon monosaccharides that play roles in many biological functions such as immune response. Neu5Ac has previously been identified as a potential biomarker for the presence and pathogenesis of cardiovascular disease (CVD), diabetes and cancer. More recent research has highlighted acetylated sialic acid derivatives, specifically Neu5,9Ac2, as biomarkers for oral and breast cancers, but advances in analysis have been hampered due to a lack of commercially available quantitative standards. We report here the synthesis of 9\u2010O\u2010 and 4\u2010O\u2010acetylated sialic acids with optimisation of previously reported synthetic routes. Neu5,9Ac2 was synthesised in 1 step in 68\u2009% yield. Neu4,5Ac2 was synthesised in 4 steps in 39\u2009% overall yield. Synthesis was followed by analysis of these standards via quantitative NMR (qNMR) spectroscopy. Their utilisation for the identification and quantification of specific acetylated sialic acid derivatives in biological samples is also demonstrated. Synthesised acetylated derivatives of N\u2010acetylneuraminic acid, Neu5,9Ac2 and Neu4,5Ac2, alongside commercially available Neu5Ac and Neu5Gc were utilised as standards for the quantitative analysis of these derivatives in plasma and serum samples. Multiple derivatives could be detected in one assay, which exhibited high specificity and low limits of detection and quantitation. Neu5Ac is a nine\u2010carbon backbone monosaccharide with a carboxylic acid functional group Figure\u2005 and is oN\u2010glycans as the terminating unit was treated with trimethyl orthoacetate in the presence of catalytic p\u2010toluene sulfonic acid for 20\u2005minutes to afford Neu5,9Ac2(2) in 68\u2009% yield after purification using ion\u2010exchange chromatography to 3.95 and 4.26\u2005ppm in (2).Neu5,9Ac, Neu5Ac was trea2(3) was based on work by Ogura et\u2005al.(3) therefore commenced with protection of the carboxylic acid group to give a methyl ester using conditions developed by Malapelle et\u2005al. employing methanol and trifluoroacetic acid to give (4) in quantitative yield.+ resin to give (5) in 65\u2009% yield. Deprotection of the methyl ester is required in the next step. The method put forth by Ogura et\u2005al. using 1\u2005M NaOH was utilised to give (6) in quantitative yield. The acetyl group was installed at the 4\u2010position using excess pyridine and acetic anhydride and the crude mixture was then purified using ion exchange chromatography using 1\u2005M formic acid as the eluent. Removal of the formic acid under reduced pressure at 35\u2009\u00b0C, as opposed to the use of lyophilisation by Ogura et\u2005al. then afforded (3) in 60\u2009% yield to 5.17\u2005ppm in the target compound (3).The route chosen to synthesise Neu4,5Ac2 and Neu4,5Ac2 in hand, the next step was to determine the purity of the standards using qNMR techniques. The method chosen for this was based on the pulse length based concentration determination (PULCON) method,20D\u221213\u2009\u00b0 ; mp. 153\u2013155\u2009\u00b0C; 1H NMR : \u03b4 4.26 , 4.07 , 4.00\u20133.90 , 3.87\u20133.77 , 3.49 , 2.19 , 2.00 , 1.94 , 1.81\u20101.72 : \u03b4 174.73 (C13), 174.41 (C11), 171.22 (C14), 95.48 (C2), 70.15 (C7), 69.2 (C8), 67.59 (C6), 66.80 (C4), 66.26 (C9), 52.03 (C5), 38.89 (C3), 22.6 (C12), 20.19 (C15) ppm; IR vmax [cm\u22121] (powder) 3295 , 1719 (C=O), 1034 (C\u2212O); HRMS (ESI): m/z calc for C13H22O10N: 352.1238 [M+H]+; found: 351.1239.Methyl 5\u2010Acetamido\u20103,5\u2010dideoxy\u2010D\u2010glycero\u2010D\u2010galactononulopyranosonate (4):N\u2010acetylneuraminic acid 1 in methanol (40\u2005mL) was added TFA . This was stirred at room temperature for 48\u2005hrs. The solvent was removed under reduced pressure to give the methyl ester 4 as a white solid [\u03b1]20D\u221226\u2009\u00b0 ; mp. 177\u2013178\u2009\u00b0C; 1H NMR : \u03b4 3.94 , 3.80 , 3.72 , 3.60 , 3.50 , 3.42 , 2.19 , 1.92 , 1.79 ppm 13C NMR : \u03b4 174.79 (C13), 171.36 (C11), 95.29 (C2), 70.29 (C6), 70.05 (C8), 68.14 (C7), 66.62 (C4), 63.10 (C9), 53.43 (C14), 52.00 (C5), 38.60 (C3), 22.00 (C12) ppm; IR vmax [cm\u22121] (powder) 3256 , 2944 (C\u2212H), 1751 (C=O), 1026 (C\u2212O); HRMS (ESI): m/z calc for C12H21O9NNa: 352.1109 [M+H]+; found: 346.1102.O\u2010isopropylidene\u2010D\u2010glycero\u2010\u03b2\u2010D\u2010galactononulopyranosonate (5)Methyl 5\u2010acetamido, 3,5\u2010dideoxy\u20108,9\u2010:4 in acetone (50\u2005mL) was added Amberlyst 15 H+ resin (1.5\u2005g) and 2,2\u2010dimethoxypropane . This was stirred for 3.5\u2005hrs at room temperature before being filtered to remove the resin. The resin was washed with acetone and the filtrate collected. After removing the solvent under reduced pressure, the resulting crude material was passed over a silica plug and eluted with ethyl acetate (4\u00d7200\u2005mL) to give acetonide 5 as a beige foam [\u03b1]20D\u221224\u2009\u00b0 ; mp. 164\u2013166\u2009\u00b0C; 1H NMR : \u03b4 8.07 , 6.71 , 4.82 , 4.75 , 4.02\u20103.94 , 3.90\u20103.86 , 3.85\u20103.78 , 3.68 , 3.56 , 3.52\u20103.49 , 2.03 , 1.88 , 1.61 , 1.26 , 1.23 ppm 13C NMR : \u03b4 179.94 (C13), 171.68 (C11), 107.22 (C15), 94.57 (C2), 75.92 (C8), 71.70 (C7), 68.38 (C6), 65.20 (C9), 64.94 (C4), 52.89 (C14), 52.12 (C5), 40.23 (C3), 26.55 (C16), 25.72 (C16), 22.55 (C12) ppm; IR vmax [cm\u22121] (powder) 3291 , 2926 (C\u2212H), 1740 (C=O), 1032 (C\u2212O); HRMS (ESI): m/z calc for C15H25O9NNa: 386.1422 [M+Na]+; found: 386.1416.O\u2010isopropylidene\u2010D\u2010glycero\u2010D\u2010galactononulopyranosonate (6)5\u2010Acetamido\u20103,5\u2010dideoxy\u20108,9\u2010:5 was dissolved in 1\u2005M NaOH (5\u2005mL) and stirred for 4\u2005hrs at room temperature. This was then diluted with water (10\u2005mL) and deionised with Amberlyst H+ resin. The mixture was filtered, the filtrate was collected and lyophilised to give a white solid 6 . [\u03b1]20D\u221225\u2009\u00b0 ; mp. 157\u2013158\u2009 \u00b0C; 1H NMR : \u03b4 4.25 , 4.20\u20104.11 , 4.07\u20103.97 , 3.95\u20103.85 , 3.62 , 2.09 , 1.46 , 1.36 ppm; 13C NMR : \u03b4 176.75 (C13), 174.43 (C11), 168.30 (C2), 109.58 (C14), 75.10 (C8), 70.59 (C7), 69.07 (C6), 66.99 (C9), 66.14 (C4), 52.16 (C5), 25.77 (C15), 24.27 (C15), 21.99 (C12) ppm; IR vmax [cm\u22121] (powder) 3282 , 1735 (C=O), 1427 (COOH) 1057 (C\u2212O); HRMS (ESI): m/z calc for C14H23O9NNa: 372.1265 [M+H]+; found: 372.1263.O\u2010acetyl\u2010D\u2010glycero\u2010D\u2010galactononulopyranosonate (3)5\u2010Acetamido\u20103,5\u2010dideoxy\u20104\u2010:6 in dry pyridine (0.5\u2005mL) under nitrogen was added acetic anhydride . This was stirred at room temperature for 18\u2005hours after which ethanol was added to remove excess acetic anhydride and solvent removed under reduced pressure by co\u2010evaporation with toluene (3\u00d7100\u2005mL). The resultant residue was dissolved in water (10\u2005mL) and passed through a column (2\u00d710\u2005cm) of DOWEX\u20101X8 formate anion exchange resin (100\u2013200\u2005mesh). The column was washed with water (3\u00d710\u2005mL) and eluted with formic acid (50\u2005mL). The formic acid was removed under reduced pressure to give a clear residue. The residue was dissolved in water (5\u2005mL) and lyophilised to give 4\u2010O N\u2010acetylneuraminic acid 3 as a white solid . [\u03b1]20D\u221234\u00b0 ; mp. 170\u2013172\u2009\u00b0C; 1H NMR : 5.22\u20105.12 , 4.18\u20103.99 , 3.76\u2010 3.72 , 3.63 , 3.50 , 2.24 , 1.95 , 1.87 ; 13C NMR : \u03b4 174.7 (C14), 173.38 (C13), 173.31 (C11), 95.24 (C2), 70.07 (C8), 70.00 (C7), 68.01 (C6), 63.09 (C4), 53.50 (C9), 49.38 (C5), 36.02 (C3), 21.85 (C12), 20.30 (C15) ppm; IR vmax [cm\u22121] (powder) 3340 , 2933 (C\u2212H), 1727 (C=O), 1019 (C\u2212O); HRMS (ESI): m/z calc for C13H22O10N: 352.1238 [M+H]+; found: 351.1239.Quantitative NMR analysis: Into an NMR tube was placed 600\u2005\u03bcL of a 5.03\u2005mMol maleic acid solution in D2O. A 1.8\u2005mg quantity of synthesised standard was dissolved in 1.8\u2005mL of D2O. This was split into three 600\u2005\u03bcL samples in order to analyse the standards in triplicate. After matching and tuning, each NMR sample was analysed on a 500\u2005MHz NMR spectrometer where a 1D 1H spectrum with water suppression was obtained with a relaxation delay of 20 seconds. The 360\u2009\u00b0 pulse was obtained for each spectrum using pulsecal. The pulse sequence used was noesypr1D. Equation\u20051 was used to determine the concentration and purity of each synthesised standard.2 and Neu4,5Ac2 standardsDMB labelling of the Neu5Ac, Neu5Gc, Neu5,9Ac: Neu5Ac, Neu5Gc, Neu5,9Ac2 and Neu4,5Ac2 standards were labelled using LudgerTagTM DMB Sialic Acid (LT\u2010KDMB\u201096). DMB labelling solution (20\u2005\u03bcL) was added to the standards. The samples were then vortexed and centrifuged followed by incubation at 50\u2009\u00b0C for 3\u2005hours. The labelling reaction was quenched by the addition of water to give a final volume of 500\u2005\u03bcL (480\u2005\u03bcL). Standard curves were prepared by performing serial dilution to create a standard curve with points: 0.01, 0.02, 0.1, 0.2, 0.5, 1.0 nmol. The procedure was carried out using a Hamilton MICROLAB STARlet Liquid Handling Robot.Sialic acid release and DMB labelling of human plasma and guinea pig serum: Release of sialic acid and DMB labelling of the samples was achieved using LudgerTagTM DMB Sialic Acid (LT\u2010KDMB\u201096). A 5 \u03bcL aliquot of each sample was dispensed into a 96\u2010well plate in triplicate. To this was added 25 \u03bcL of 2\u2005M acetic acid. The sample was vortexed and centrifuged followed by incubation at 80\u2009\u00b0C for 2\u2005hours. The sample was allowed to cool to room temperature and a 5 \u03bcL aliquot of each sample was transferred to a new 96\u2010well plate. To this was added 20\u2009\u03bcL of DMB labelling solution. The sample was then vortexed and centrifuged followed by incubation at 50\u2009\u00b0C for 3\u2005hours. The labelling reaction was quenched by the addition of water to give a final volume of 500\u2005\u03bcL (475\u2005\u03bcL). The samples were then subjected to a 1 in 10 dilution . All procedures, except the dispensing of the samples on the plate, were conducted using a Hamilton MICROLAB STARlet Liquid Handling Robot.Sialic acid release and DMB labelling of porcine and ovine serum: Release of sialic acid and DMB labelling of the samples was achieved using LudgerTagTM DMB Sialic Acid (LT\u2010KDMB\u201096). A stock solution was first prepared by dissolving 10\u2005mg of porcine or ovine serum in 1\u2005mL ultra\u2010pure water. A 10 \u03bcL aliquot taken from the solution of each sample was dispensed into a 96\u2010well plate in triplicate. To this was added 25\u2005\u03bcL of 2\u2005M acetic acid. The sample was vortexed and centrifuged followed by incubation at 80\u2009\u00b0C for 2\u2005hours. The sample was allowed to cool to room temperature and a 5 \u03bcL aliquot of each sample was transferred to a new 96\u2010well plate. To this was added 20\u2005\u03bcL of DMB labelling solution. The sample was then vortexed and centrifuged followed by incubation at 50\u2009\u00b0C for 3\u2005hours. The labelling reaction was quenched by the addition of water to give a final volume of 500\u2005\u03bcL (475\u2005\u03bcL). The samples were then subjected to a 1 in 10 dilution . All procedures, except the dispensing of the samples on the plate, were carried out using a Hamilton MICROLAB STARlet Liquid Handling Robot.2Sialic acid release and DMB labelling of plasma for Neu5,9Ac: Release of Sialic Acid and DMB labelling of the samples was achieved using LudgerTagTM DMB Sialic Acid (LT\u2010KDMB\u201096). A 25 \u03bcL aliquot of each sample was dispensed into a 96\u2010well plate in triplicate. To this was added 75 \u03bcL of 2.666\u2005M acetic acid. The sample was vortexed and centrifuged followed by incubation at 80\u2009\u00b0C for 2\u2005hours. The sample was allowed to cool\u2010room temperature and a 20 \u03bcL aliquot of each sample was transferred to a new 96\u2010well plate. To this was added 20\u2005\u03bcL of DMB labelling solution. The sample was then vortexed and centrifuged followed by incubation at 50\u2009\u00b0C for 3\u2005hours. The labelling reaction was quenched by the addition of water to give a final volume of 500\u2005\u03bcL (475\u2005\u03bcL). The sample was subjected to filtration through a Ludger Clean Protein Binding Membrane filtration plate (LC\u2010PBM\u2010plate) to remove excess protein. All procedures, except the dispensing of the samples on the plate and LC\u2010PBM\u2010plate filtration, were carried out using a Hamilton MICROLAB STARlet Liquid Handling Robot.Fluorescence analysis of DMB labelled sialic acid derivatives by LC\u2010fluorescence detection: DMB labelled sialic acid derivatives were analysed by LC\u2010FLD. A 5\u2009\u03bcL aliquot of each sample was injected to a LudgerSep\u2122 uR2 UHPLC column at 30\u2009\u00b0C on a Dionex UltiMate\u21223000 RSLCnano system with a fluorescent detector . For Neu5Ac analysis, an isocratic solvent system was used (7\u2009:\u20099\u2009:\u200984 MeOH : ACN : H2O) for 15 minutes including an ACN wash. For Neu5,9Ac2 analysis a variable solvent system was used: 7\u2009:\u20096\u2009:\u200987 MeOH:ACN:H2O for 6.5 minutes followed by 6\u2009:\u20099\u2009:\u200985 MeOH:ACN:H2O for 11.5\u2005minutes. Integration of resultant peaks was performed using Chromeleon 7. LOD and LOQ regression analysis and calculation was performed using Microsoft Excel 2019.1As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "The bridging O2\u2212 ligand is located on a twofold rotation axis, resulting in point group 2 for the entire complex. The MnIII atom is displaced out of the 24-atom mean plane of the porphyrine entity by 0.52\u2005\u00c5. C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions help to stabilize the mol\u00adecular packing within the crystal structure.In the crystal structure of the title oxido-bridged binuclear complex, [Mn The two MnIII ions are displaced by 0.48 and 0.52\u2005\u00c5 from their respective 24-atom mean plane. It is inter\u00adesting to note that the \u03bc-oxido species [Mn(OEP)]2O is very unstable in halocarbon solvents . Figs. 12O are bridged by a single oxido ligand with an Mn\u2014O distance of 1.7600\u2005(3)\u2005\u00c5 and an Mn\u2014O\u2014Mn bridging angle of 176.1\u2005(2)\u00b0. The Mn1\u22efMn1\u2032 separation 2(OH)}ClO4, the title compound shows virtually the same metal displacement from the 24-atom mean plane (0.52\u2005\u00c5), while a larger Mn\u2014O\u2014Mn bridging angle [176.1\u2005(2) versus. 160.4\u2005(8)\u00b0] and a shorter Mn\u2014O distance [1.7600\u2005(3) versus. 2.026\u2005(1)\u2005\u00c5] is observed.In the crystal structure of the title complex, the asymmetric unit contains one deprotoanted porphyrin mol\u00adecule located in general position and an oxygen atom on a twofold rotation axis ]Cl were prepared according to literature protocols ]2O (10\u2005mg) was dissolved in 4\u2005ml of chloro\u00adbenzene and cannula-transferred into 8\u2005mm glass tubes, then carefully layered with hexa\u00adnes before sealing the tubes. X-ray quality crystals were obtained several weeks later.The title compound was prepared following a reported procedure I. DOI: 10.1107/S2414314622008690/wm4172Isup2.hklStructure factors: contains datablock(s) I. DOI: 2204345CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Longer N\u2014H\u22efO hydrogen bonds link the tetra\u00admers into [010] chains.In the title hydrated Schiff base, the dihedral angle between the aromatic rings is 5.06\u2005(11)\u00b0 and an intra\u00admolecular O\u2014H\u22efN hydrogen bond closes an S(6) ring. In the crystal, O 13H12N4O3\u00b7H2O, the dihedral angle between the aromatic rings is 5.06\u2005(11)\u00b0 and an intra\u00admolecular O\u2014H\u22efN hydrogen bond closes an S(6) ring. In the crystal, Ow\u2014H\u22efO and Ow\u2014H\u22efN (w = water) hydrogen bonds link the components into centrosymmetric tetra\u00admers (two Schiff bases and two water mol\u00adecules). Longer N\u2014H\u22efO hydrogen bonds link the tetra\u00admers into [010] chains. A weak C\u2014H\u22efO hydrogen bond and aromatic \u03c0\u2013\u03c0 stacking between the pyrazine and phenyl rings [centroid\u2013centroid separations = 3.604\u2005(2) and 3.715\u2005(2)\u2005\u00c5] are also observed.In the title hydrated Schiff base, C Their applications include mol\u00adecular switches (Coskun S(6) ring. The C7\u2014N2 bond length [1.278\u2005(3)\u2005\u00c5] is consistent with a normal carbon\u2013nitro\u00adgen double bond. In the crystal, Ow\u2014H\u22efO and Ow\u2014H\u22efN (w = water) hydrogen bonds link the components into centrosymmetric tetra\u00admers (two Schiff base and two water mol\u00adecules). Longer N\u2014H\u22efO hydrogen bonds link the tetra\u00admers into [010] chains \u00b0 and an intra\u00admolecular O2\u2014H2\u22efN2 hydrogen bond closes an s Table\u00a01. The pacPyrazine-2-carbohydrazide was reacted with 2-hy\u00addroxy-3-meth\u00adoxybenzaldehyde under reflux in 25\u2005ml methanol for 8\u2005h. After cooling and solvent removal by rotary evaporation, a light yellow solid was obtained, which was recrystallized from methanol solution at room temperature to obtain colourless crystals of the title compound.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314619017310/hb4335sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314619017310/hb4335Isup2.hklStructure factors: contains datablock(s) I. DOI: 1973543CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Orientogalba ollula has been reported as intermediate hosts of many zoonotic trematodes. Here, we investigated the prevalence of zoonotic trematodes within O. ollula in Guangxi, China, and assessed their zoonotic potential.Food-borne parasitic diseases decrease food safety and threaten public health. The snail species is an intermediate host for numerous human parasitic trematodes. Echinostoma revolutum was recorded by dissecting infected ducklings from 1-day post infection (dpi) to 10 dpi.Snails were collected from 54 sites in 9 cities throughout Guangxi. The snail and trematode larvae species were determined by combining morphological characteristics and molecular markers. The trematodes prevalence and constituent ratio were calculated and compared among different habitat environments. Phylogenetic trees of the trematode species were constructed using the neighbor-joining method with nuclear internal transcribed spacer 2 (ITS2) sequences. The developmental cycles of the isolated trematodes were examined by experimental infection in ducks. The developmental characteristics of O. ollula from 11 sample sites. Morphological together with molecular identification, showed that E. revolutum, Australapatemon sp., Hypoderaeum conoideum, Pharyngostomum cordatum, and Echinostoma sp. parasitized O. ollula, with the highest infection rate of E. revolutum (13.0%). However, no Fasciola larvae were detected. The trematodes prevalence and constituent ratio varied in two sub-biotypes (P\u2009<\u20090.01). A neighbor-joining tree analysis of ITS2 sequences resulted in distinct monophyletic clades supported by sequences from isolated larvae with high bootstrap values. Ducklings exposed to O. ollula infected with Echinostoma sp., E. revolutum, and H. conoideum larvae were successfully infected. The animal model for Echinostoma revolutum was successfully established. E. revolutum matured from larvae to adult at 10 dpi in the intestine of the duck, and the developmental characteristics of E. revolutum were characterized by the maturation of the reproductive and digestive organs at 6\u20138 dpi.The overall prevalence of trematode larvae was 22.1% (1818/8238) in O. ollula from Guangxi, China. Existing trematodes infection in animals and human clinical cases, coupled with the wide geographical distribution of O. ollula, necessitate further evaluations of the potential risk of spillover of zoonotic infection from animal to human and vice versa.This study revealed a high prevalence of zoonotic trematodes in The online version contains supplementary material available at 10.1186/s40249-022-01014-7. Corresponding trematodes belong to the families Fasciolidae, Paragonimidae and Echinostomatidae in the feces, and the morphological characteristics of adult Echinostoma sp. were presented as measurements: body length (9.8\u2009\u00b1\u20091.8\u00a0mm), width (1.2\u2009\u00b1\u20092.3\u00a0mm), oral sucker [(638.9\u2009\u00b1\u20091.9)\u2009\u00d7\u2009(399.2\u2009\u00b1\u200915.8) \u03bcm], acetabulum [(1591.2\u2009\u00b1\u200917.8)\u2009\u00d7\u2009(1338.2\u2009\u00b1\u200919.6) \u03bcm], pharynx [(492.6\u2009\u00b1\u200954.3)\u2009\u00d7\u2009(331.7\u2009\u00b1\u200974.8) \u03bcm], anterior testis [(1120.5\u2009\u00b1\u2009139.6)\u2009\u00d7\u2009(707.4\u2009\u00b1\u200949.2) \u03bcm], posterior testis [(1274.9\u2009\u00b1\u2009181.1)\u2009\u00d7\u2009(880.4\u2009\u00b1\u200949.2) \u03bcm], and ovary [(818.9\u2009\u00b1\u200910.2)\u2009\u00d7\u2009(527.9\u2009\u00b1\u200948.3) \u03bcm].We conducted an infection experiment for three kinds of trematode to evaluate the rates of parasite establishment in ducklings. Ducklings were individually exposed to H. conoideum eggs in ducklings fed with infected O. ollula from three sites after a median duration of 12 dpi (range: 9 dpi to 14 dpi). The morphological characteristics of adult H. conoideum were as follows: body length (1.05\u2009\u00b1\u20090.61\u00a0mm), width (1.5\u2009\u00b1\u20090.27\u00a0mm), oral sucker [(424.2\u2009\u00b1\u200916.1)\u2009\u00d7\u2009(293\u2009\u00b1\u200911.2) \u03bcm], acetabulum [(1610.2\u2009\u00b1\u200998.4)\u2009\u00d7\u2009(1594.6\u2009\u00b1\u2009198.7) \u03bcm], pharynx [(379.8\u2009\u00b1\u200953.2)\u2009\u00d7\u2009(253.6\u2009\u00b1\u200944.9) \u03bcm], anterior testis [(1902.6\u2009\u00b1\u2009164.6)\u2009\u00d7\u2009(875.3\u2009\u00b1\u2009140.8) \u03bcm], posterior testis [(2045.2\u2009\u00b1\u2009255.8)\u2009\u00d7\u2009(898.2\u2009\u00b1\u2009139.2) \u03bcm], ovary [(751.7\u2009\u00b1\u2009129.3)\u2009\u00d7\u2009(553.9\u2009\u00b1\u2009110.6) \u03bcm], and the possession of 50 spines. E. revolutum eggs [(104.1\u2009\u00b1\u200915.1)\u2009\u00d7\u2009(63.1\u2009\u00b1\u200912.6\u00a0\u03bcm] were found on 10 dpi. The morphology of the adult E. revolutum was characterized by: body length (8.5\u2009\u00b1\u20091.2\u00a0mm), width (2.2\u2009\u00b1\u20090.7\u00a0mm), oral sucker [(260.0\u2009\u00b1\u200932.5)\u2009\u00d7\u2009(180.1\u2009\u00b1\u200919.8) \u03bcm], acetabulum [(741.6\u2009\u00b1\u200923.7)\u2009\u00d7\u2009(598.3\u2009\u00b1\u200925.1) \u03bcm], pharynx [(193.1\u2009\u00b1\u200930.5)\u2009\u00d7\u2009(150.6\u2009\u00b1\u200927.8) \u03bcm], anterior testis [(628.0\u2009\u00b1\u200931.2)\u2009\u00d7\u2009(459.4\u2009\u00b1\u200927.8) \u03bcm], posterior testis (725.5\u2009\u00b1\u200917.1\u2009\u00d7\u2009557.9\u2009\u00b1\u200918.7\u00a0\u03bcm), ovary [(411.8\u2009\u00b1\u200918.5)\u2009\u00d7\u2009(311.2\u2009\u00b1\u200915.4) \u03bcm], and the presence of a head collar with 37 spines.In contrast, we found E. revolutum in duckling hosts, because there were insufficient snails infected with metacercariae of Echinostoma sp. and H. conoideum. The developmental characteristics of E. revolutum were recorded by dissecting infected ducklings from 1 to 10 dpi when eggs in the feces were first detected. E. revolutum could be obtained in the small intestine from 1 to 7 dpi and then migrate and reside in the cecum and colon from 8 to 10 dpi. The body length developed from 490\u00a0\u03bcm to 8500.5\u00a0\u03bcm (a 17-fold increase). At 1 dpi, juveniles had a circumoral collar bearing 37 spines in a double circle and characterized by clearly visible oral suckers, acetabulum, pharynx, esophagus, and cecum. At 1 dpi, the tiny structure of the testis appeared. By 4 dpi, the ovaries were beginning to organize and develop, and the seminal receptacle began to form. The tubular-shaped uterus loomed at 4 dpi, and maturation of the reproductive and digestive organs occurred at 6 to 8 dpi. The vitelline glands were the last to appear, and several eggs deposited in the uterus were observed at 9 dpi. E. revolutum larvae matured at 10 dpi and excreted eggs ."} +{"text": "Aqua\u00adtri\u00adfluorido\u00adboron and ethyl\u00adene carbonate form a 1:2 co-crystal with a C=O\u22efH\u2014O\u2014H\u22efO=C hydrogen-bonding motif. 3H2O\u00b72OC(OCH2)2, was determined by low-temperature single-crystal X-ray diffraction. The co-crystal crystallizes in the ortho\u00adrhom\u00adbic space group P212121 with four formula units per unit cell. The asymmetric unit consists of an aqua\u00adtri\u00adfluorido\u00adboron mol\u00adecule and two ethyl\u00adene carbonate mol\u00adecules, connected by O\u2014H\u22efO=C hydrogen bonds. This crystal structure is an inter\u00adesting example of a superacidic BF3H2O species co-crystallized with an organic carbonate.The crystal structure of the co-crystal of aqua\u00adtri\u00adfluorido\u00adboron with two ethyl\u00adene carbonate mol\u00adecules, BF The overall shape of the BF3 moiety in BF3H2O in terms of bond lengths and angles is similar to that of the BF4\u2212 anion graph-set motif 2 co-crystal were discovered when a crystalline sample of the air-sensitive BF3\u00b7OC(OCH2)2 adduct was examined under a protective cold nitro\u00adgen stream at about \u221250\u2005\u00b0C. The BF3\u00b7OC(OCH2)2 compound was synthesized from dry ethyl\u00adene carbonate and BF3 gas under anhydrous conditions, as described previously 2 were located in a droplet at the tip of the aluminium trough I. DOI: 10.1107/S2414314623000627/wm4182Isup2.hklStructure factors: contains datablock(s) I. DOI: 2237804CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "D2 symmetry. They show a largely twisted double bond, the mol\u00adecular halves enclosing dihedral angles of 62.86\u2005(4) and 61.22\u2005(3)\u00b0.The asymmetric unit of the title compound contains two almost identical mol\u00adecules, which have approximate 42H24, contains two almost identical mol\u00adecules. The mol\u00adecules have approximate D2 symmetry. They show a largely twisted double bond, the mol\u00adecular halves enclosing dihedral angles of 62.86\u2005(4) and 61.22\u2005(3)\u00b0. The crystal studied was twinned by non-merohedry.The asymmetric unit of the title compound, C The dihedral angle between these planes is 62.86\u2005(4)\u00b0 and the length of the central C1A\u2014C22A bond is 1.408\u2005(5)\u2005\u00c5. The values for mol\u00adecule B are very similar: maximum deviations from the mean planes are \u22120.293\u2005(4)\u2005\u00c5 for C5 (B1) and 0.170\u2005(4)\u2005\u00c5 for C26 (B2). The dihedral angle amounts to 61.22\u2005(3)\u00b0 and the length of the central C1B\u2014C22B bond is 1.395\u2005(5)\u2005\u00c5. Whereas the length of the twisted and elongated bond connecting the two halves of mol\u00adecules A and B correlates very well with the calculated value of 1.408\u2005\u00c5, the calculated torsion angle of 52\u00b0 is about 10\u00b0 too small. The solid material gives a very weak single ESR signal.Benzoannulated fulvenes attract chemists because of their electronic properties I, global. DOI: 10.1107/S2414314622001699/bt4120Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622001699/bt4120Isup3.cmlSupporting information file. DOI: 2151512CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The different intra and inter\u00admolecular hydrogen-bonding inter\u00adactions in the crystal structure of the title salt are discussed. 11H17N2O+\u00b7C7H3N2O7\u2212, exhibits secondary nitro\u00adgen atoms (N\u2014H) in the 2-meth\u00adoxy\u00adphenyl\u00adpiperazine (2MeOPP) cation, which is protonated with a phenolic hydrogen atom of 3,5-di\u00adnitro\u00adsalicylic acid (DNSA). One of the oxygen atoms of the nitro group in the 3,5-di\u00adnitro\u00adsalicylate anion is disordered over two orientations with occupancy factors of 0.65\u2005(7) and 0.35\u2005(7) . The 2-meth\u00ad\u00adoxy\u00adphenyl\u00adpiperazinium cation and 3,5-di\u00adnitro\u00adsalicylate anion are linked in the asymmetric unit by a bifurcated N\u2014H\u22efO hydrogen bond, which formed is between the H atom in the protonated piperazinium unit of the cation and the carb\u00adoxy\u00adlic acid group in the anion. The piperazine ring adopts a chair conformation. The crystal structure features N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds inter\u00adactions, which lead to the formation of a sandwich-like arrangement. Hirshfeld surface analysis was used to determine the relative contributions of various inter\u00admolecular inter\u00adactions, indicating that that H\u22efO/O\u22efH (38. 3%) and H\u22efH (31. 8%) contacts are the major contributors.The title salt , C Piperazine and substituted piperazine derivatives are often used as inter\u00admediates for a wide range of pharmaceuticals, polymers, dyes, corrosion inhibitors and surfactants. In particular, (2-meth\u00adoxy\u00adphen\u00adyl)piperazine derivatives are used as 5-HT1A receptor ligands with reduced \u03b11-adrenergic activity -tartaric acid] salts and their supra\u00admolecular features have been reported piperazine is a substituted cyclo aliphatic amine with two nitro\u00adgen atoms at opposite positions of the six-membered ring. A substituent 2-meth\u00adoxy\u00adphenyl group is attached to one of the nitro\u00adgen atoms while the other has one attached hydrogen atom , 3,5-di\u00adnitro-2-oxidobenzoate and piperazine, we have now investigated the crystal structure of 1-(2-meth\u00adoxy\u00adphen\u00adyl) piperazinium 3,5-dinitro\u00adsalicylate (I)2.P\u012b with the asymmetric unit comprising one 2-meth\u00adoxy\u00adphenyl\u00adpiperazinium (2MeOPP)1+ cation and one 3,5-di\u00adnitro\u00adsalicylate (DNSA)1\u2212 anion \u2005\u00c5, \u03b8 = 176.3\u2005(3)\u00b0, \u03c6 = 338\u2005(4)\u00b0. One of the oxygen atoms of the nitro group (atom O4) in the 3,5-di\u00adnitro\u00adsalicylate anion is disordered over two orientations with occupancy factors of 0.65\u2005(7) and 0.35\u2005(7). Both nitro groups, the phen\u00adoxy\u00adlate oxygen atom and a carb\u00adoxy\u00adlic acid group in the anion are coplanar with an r.m.s. deviation of 0.0074\u2005\u00c5. A bifurcated inter\u00admolecular N\u2014H\u22efO hydrogen bond [N3\u2014H3A\u22efO5 = 2.936\u2005(3)\u2005\u00c5 and N3\u2014H3A\u22efO6 = 3.153\u2005(3)\u2005\u00c5] links the cation and anion in the asymmetric unit.The title salt crystallizes in the triclinic space group on Fig.\u00a01. The pipK\u03b1 COOH = 2.2) is easier than that of the phenolic \u2013OH group (pK\u03b1 OH = 6.8). 62 carboxyl\u00adate moiety structures (COO\u2212) and 70 phenolate anion structures (O\u2212) were found in a search of the Cambridge Structural Database \u2005\u00c5, which also indicates that the strong intra\u00admolecular hydrogen bond between the O6 and O7 atoms. Similar types of intra\u00admolecular hydrogen bonds were observed in salicylic acid with a distance of 2.62\u2005\u00c5 inter\u00adaction, which links two neighbouring cations and anions into a centrosymmetric tetra\u00admeric architecture, which is further stabilized by the C14\u2014H14\u22efO5v inter\u00adaction [3.481\u2005(3)\u2005\u00c5] and yields a macrocyclic ring structure with an ii inter\u00adaction [3.581\u2005(3)\u2005\u00c5], which links two neighbouring (DNSA)1\u2212 units with an 1\u2212 units are further linked through the previously mentioned bifurcated N3\u2014H3A\u22ef inter\u00adaction and the N3\u2014H3B\u22efO7i [2.787\u2005(3)\u2005\u00c5], C10\u2014H10A\u22efO4A [3.118\u2005(10)\u2005\u00c5] inter\u00adactions into a layered structure propagating parallel to the b axis , and N3\u2014H3B\u22efO7], the N3\u2014H3B\u22efO7 inter\u00adaction is stronger [D\u22efA = 2.787\u2005(3)\u2005\u00c5] than the other two, which is due to the fact that two charged components are involved in this inter\u00adaction, i.e. the phenolate O7 atom in DNSA\u22121 and the protonated N3\u2014H3B unit in 2MeOPP+1. All of the above inter\u00adactions facilitate the arrangement of the DNSA1\u2212 ions in a layered mol\u00adecular structure. The top and bottom sides of the DNSA1\u2212 layers are stabilized by the two adjacent cationic layers. As a result, a sandwich-like arrangement is observed. Briefly, the layered DNSA1\u2212 units form the core with the top and bottom sides of the cation layers arranged facing. An overall packing diagram is shown Fig.\u00a06The oxygen atoms in both nitro groups (O1\u2013O4), the carb\u00adoxy\u00adlic acid group (O5 and O6) and a phenolate moiety (O7) in the DNSA anion all act as acceptors for various inter\u00admolecular N\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions, except for atom O4 Table\u00a01. In the if Fig.\u00a03. Atom O1if Fig.\u00a04. Neighbois Fig.\u00a05. Of the 4.Crystal Explorer 17.5 are shown in Fig.\u00a07A, O7, O6, H10B and H3B suggest that these atoms participate in hydrogen-bonding inter\u00adactions (see Table\u00a01de + di = 1.8\u2005\u00c5] make the most significant contribution (38.3%), followed by H\u22efH contacts [symmetrical blunt spikes at de + di = 2.4\u2005\u00c5], which contribute 31.8%, while C\u22efH, N\u22efH, C\u22efO, O\u22efN, C\u22efN and C\u22efC contacts contribute 11.6%, 1.7%, 6.7%, 2.7%, 1.9%, 0.5% and 2.8%, respectively. Other significant peaks for various non-covalent contacts are indicated in the FP plot piperazine and 3,5-di\u00adnitro\u00adsalicylic acid in an equimolar ratio. The stoichiometrically (1\u2005mmol) weighed starting materials were completely dissolved in 50\u2005mL of methanol at room temperature and stirred continuously for 3\u2005h. The homogeneous solution was filtered using Whatmann filter paper and placed in a dust-free atmosphere, and allowed to evaporate slowly at room temperature. A suitable single crystal was harvested after a growth period of 25 days.7.Uiso(H) = 1.2Ueq(C)].Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022006831/ex2058sup1.cifCrystal structure: contains datablock(s) I, publication_text. DOI: 2183987CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Inter\u00admolecular N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds form a three-dimensional network in the crystal, connecting mol\u00adecules through the O atoms of solvent mol\u00adecules. 16H11BrN6O\u00b72C2H6OS, the 1,2,3,7-tetra\u00adhydro\u00adimid\u00adazopyridine ring system and the oxindole moiety are both nearly planar and their planes form a dihedral angle of 86.04\u2005(5)\u00b0 with each other. Inter\u00admolecular N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds link mol\u00adecules in the crystal through the O atoms of the solvent mol\u00adecules, generating a three-dimensional network. A Hirshfeld surface analysis was performed to further analyse the inter\u00admolecular inter\u00adactions.In the title compound, C These ring systems make a dihedral angle of 86.04\u2005(5)\u00b0 with each other. The cyano (\u2013C\u2261N) and amine (NH2) groups form an inter\u00admolecular hydrogen bond with one dimethyl sulfoxide (DMSO) group, giving an S(10) motif (see Scheme3.A\u2014N1 bond (which has some multiple-bond character) acts as an electron donor to Br1 in a kind of \u2018halogen bond\u2019, with a Br1\u22efC8A distance of 3.284\u2005(2)\u2005\u00c5.In the crystal, mol\u00adecules are linked through the O atoms of the DMSO solvent mol\u00adecules by inter\u00admolecular N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds which, together with C\u2014H\u22efN hydrogen bonds, form a three-dimensional (3D) network Table\u00a01. The \u03c0-cCrystalExplorer plotted over the range from \u22120.6206 to 1.3180 a.u. The inter\u00adactions given in Table\u00a01The Hirshfeld surfaces were calculated and the two-dimensional (2D) fingerprint plots generated using a), and those delineated into H\u22efH, N\u22efH/H\u22efN, O\u22efH/H\u22efO, C\u22efH/H\u22efC and Br\u22efH/H\u22efBr contacts are shown in Figs.\u00a04b)\u2013(f). The percentage contributions to the Hirshfeld surfaces from the various inter\u00adatomic contacts are as follows: H\u22efH , N\u22efH/H\u22efN , O\u22efH/H\u22efO , C\u22efH/H\u22efC and Br\u22efH/H\u22efBr . Other minor contributions to the Hirshfeld surface are from Br\u22efC/C\u22efBr (3.9%), Br\u22efN/N\u22efBr (2.0%), C\u22efC (1.5%), S\u22efC/C\u22efS (0.8%), S\u22efH/H\u22efS (0.6%), S\u22efN/N\u22efS (0.4%), O\u22efN/N\u22efO (0.4%) and Br\u22efO/O\u22efBr (0.3%).The overall 2D fingerprint plot for (1) is given in Fig.\u00a044.et al., 2016H-indol-2-one unit of (1) gave 87 hits. The three compounds most resembling (1) are (I) , N\u2014H\u22efO hydrogen bonds lead to the formation of chains along the c-axis direction. Within the chains there are further N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds enclosing via N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds involving the dimethyl sulfoxide solvent mol\u00adecule which acts as both an acceptor and a donor.In the crystal of (II), the asymmetric unit contains two independent mol\u00adecules (A and B) having similar conformations. In the crystal, mol\u00adecules are linked by N\u2014H\u22efO hydrogen bonds, forming chains along the a axis which enclose In (III), two intra\u00admolecular N\u2014H\u22efO hydrogen bonds are formed, each closing an S(6) loop. In the crystal, strong N\u2014H\u22efO hydrogen bonds lead to the formation of zigzag chains along the c axis. These are consolidated in the 3D crystal packing by weak N\u2014H\u22efO hydrogen bonding, as well as by C\u2014H\u22efO, C\u2014H\u22efBr and C\u2014H\u22ef\u03c0 inter\u00adactions.In solution . Single crystals of (1) were grown from DMSO solution.To a solution of 2-(5-bromo-2-oxoindolin-3-yl\u00adidene)malono\u00adnitrile , which was previously prepared by a known procedure : \u03b4 3.50 , 6.61 , 6.78 , 7.35 , 7.37 , 7.73 , 10.44 . 13C NMR : \u03b4 42.46 (CH2N), 45.15 (CH2N), 51.24 (Cquat), 51.71 (=Cquat), 54.69 (=Cquat), 112.02 (CHarom), 114.43 (Br\u2014Carom), 119.63 (CN), 120.15 (CN), 128.02 (CHarom), 131.90 (CHarom), 137.83 (Carom), 140.80 (Carom), 152.19 (=Cquat), 154.76 (=Cquat), 179.67 (O=C).6.Uiso(H)\u00a0= 1.2Ueq(N), and C\u2014H\u00a0= 0.95\u20130.99\u2005\u00c5 with Uiso(H)\u00a0= 1.2 or 1.5Ueq(C). Both DMSO solvent mol\u00adecules are disordered over two positions, with final occupancies of 0.90:0.10 for the first and 0.95:0.05 for the second mol\u00adecule. In the first disordered DMSO molecule, the C17B and C18B atoms of the minor component were refined isotropically. The disordered atoms O2A/O2B, O3A/O3B, C19A/C19B and C20A/C20B were refined with anisotropic displacement parameters, constrained to be the same for both components. The S\u2014C and S\u2014O bond lengths in both disordered DMSO mol\u00adecules were restrained to similarity.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022004741/zv2013sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989022004741/zv2013Isup2.hklStructure factors: contains datablock(s) I. DOI: 2170241CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The correct data result is \u2018IBL clear presentation time (23.59 \u00b1\u20094.47 vs. 1026.80 \u00b1\u2009318.34 s) (p <\u20090.001)\u2019 instead of \u2018IBL clear presentation time (23.6 \u00b1\u20094.4 vs. 23.6 \u00b1\u20094.4 s) (p <\u20090.01)\u2019. The results section should read as below:Results: An IBL was visible in 98% of patients in the ICGF\u2010based group, even with low doses of ICG. The ICGF\u2010based group was significantly associated with a shorter IBL clear presentation time (23.59\u2009\u00b1\u20094.47\u2009vs.\u20091026.80\u2009\u00b1\u2009318.34 s) (p\u2009<\u20090.001) and operative time (89.3\u2009\u00b1\u200931.6 vs. 112.9\u2009\u00b1\u200933.3\u2009min) (p\u2009<\u20090.01) compared to the MID group. The incidence of postoperative prolonged air leaks was higher in the MID group than in the ICGF\u2010based group . There were no significant differences in bleeding volume, chest tube duration, postoperative hospital stays, surgical margin width, and other postoperative complications.The authors apologize for the error and any inconvenience it may have caused."} +{"text": "The title compound features a B\u2014O\u2014B bond angle of 132.75\u2005(13)\u00b0. 40H36B2N4O5, the B\u2014O\u2014B bond angle is 132.75\u2005(13) and the dihedral angle between the benzodiazborole rings is 73.02\u2005(5)\u00b0. In the crystal, weak C\u2014H\u22efO inter\u00adactions link the mol\u00adecules.In the title compound, C Additionally, it is likely that a p-type electronic inter\u00adaction exists between O1 and the adjacent boron atoms (B1A and B1B) that would serve to open up the bond angle substanti\u00adally beyond the textbook angle of 109.5\u00b0 for an O atom bearing two lone pairs of electrons. As a result of steric encumbrance, the B1A and B1B benzodi\u00adaza\u00adborole rings are angled away from one another to a near perpendicular orientation, with a plane-to-plane tilt of 73.02\u2005(5)\u00b0. The dihedral angles between the B1A benzodi\u00adaza\u00adborole ring system and its pendant p-meth\u00adoxy\u00adbenzene rings are 80.49\u2005(6) and 49.84\u2005(7)\u00b0 for the C7A and C14A rings, respectively. Comparable data for the B1B ring system and its pendant C7B and C14B rings are 78.32\u2005(6) and 65.96\u2005(7)\u00b0, respectively. The C atoms of the meth\u00adoxy groups are all close to their respective ring planes: C13A [deviation = 0.333\u2005(2)\u2005\u00c5]; C20A [0.254\u2005(2)\u2005\u00c5]; C13B [\u22120.040 (2\u2005\u00c5)]; C20B [0.193\u2005(2)\u2005\u00c5].The B1In the crystal, weak C\u2014H\u22efO inter\u00adactions Table\u00a01 link theN1,N2-bis\u00ad(4-meth\u00adoxy\u00adphen\u00adyl)benzene-1,2-di\u00adamine , was obtained in 87% yield. The single-crystal X-ray structure of the di\u00adaza\u00adborole chloride has been deposited with the Cambridge Structural Database . After stirring overnight, a white precipitate of the tri\u00adethyl\u00adammonium chloride formed that was then filtered and discarded. The filtrate was dried under reduced pressure to give the crude product. The solid was extracted in a fritted glass filter with a minimum volume of benzene, and the filtrate was evaporated under reduced pressure to give the title compound in 85% yield.Under an anhydrous nitro\u00adgen atmosphere, a solution was prepared that contained 3.0\u2005mmol of (II), four equivalents of tri\u00adethyl\u00adamine, and \u223c200\u2005ml of 1,2-di\u00admeth\u00adoxy\u00adethane. This solution was then treated with half an equivalent of water global, I. DOI: 10.1107/S2414314620012481/hb4361Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314620012481/hb4361Isup4.cmlSupporting information file. DOI: 2031384CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit is composed of half a binuclear silver complex located about a center of symmetry, a potassium cation and 2.6 disordered water mol\u00adecules. The whole binuclear silver complex is generated by inversion symmetry with the pyrazine ring being located about an inversion centre. The ligand coordinates in a bis-tetra\u00addentate manner. The binuclear silver complex anions are linked via bridging Ag\u22efS\u22efAg zigzag bonds, forming a network lying parallel to the bc plane. The networks are linked by Ocarboxyl\u00adate\u22efK+\u22efOcarboxyl\u00adate bridging bonds to form a framework. The disordered water mol\u00adecules are present near to the K+ cations.The reaction of AgNO In I this value is 2.550\u2005(5)\u2005\u00c5. For Ag\u2014Ocarboxyl\u00adate there were over 2,800 hits with the bond lengths varying from 1.967 to 3.089\u2005\u00c5 . In I the Ag\u2014Ocarboxyl\u00adate bond lengths are almost equal; 2.470\u2005(5) and 2.466\u2005(6)\u2005\u00c5. Finally for the Ag\u2014S(CH2)2\u2014 bond-length type there were over 1,000 hits with the bond length varying from 2.361 to 3.583\u2005\u00c5 . In I the Ag\u2014S(CH2)2\u2013 bond lengths vary from 2.604\u2005(2) to 2.926\u2005(2)\u2005\u00c5, both values involve the bridging atom S1, while distance Ag1\u2014S2ii is 2.824\u2005(2)\u2005\u00c5 \u2212x, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01].The three chelate rings are far from flat, as indicated by the torsion angles given in Table\u00a01K+\u22efOcarboxyl\u00adate bonds lengths vary from 2.608\u2005(6) to 2.751\u2005(6)\u2005\u00c5, and there is one weak contact K1\u22efO1 at 3.289\u2005(6)\u2005\u00c5 potassium] potassium] to 2.8668\u2005(13)\u2005\u00c5 in UBUPAK and from 2.8197\u2005(14) to 3.0449\u2005(15)\u2005\u00c5 in MUMPIW. In UBUPAK the K+ cation has a coordination number of 8 (KO8) and a distorted dodeca\u00adhedral geometry, while in MUMPIW the K+ ion has a coordination number of 7 (KO6N) and has an edge-sharing penta\u00adgonal anti\u00adprism geometry. In I, the stronger K\u22efO bond lengths are shorter and, owing to the presence of the disordered water mol\u00adecules, it is not clear what the K+ ion coordination number or geometry are.Selected bond lengths and bond angles involving atom K1 are also given in Table\u00a01\u2005\u00c5 Fig.\u00a03. A searcI, the networks of the binuclear silver complex anions are linked by the bridging Ocarboxyl\u00adate\u22efK+\u22efOcarboxyl\u00adate bonds to form a framework ] tetra\u00adkis\u00ad(sulfanedi\u00adyl)}tetra\u00adacetato)}-bis\u00ad[silver(I)]-bis\u00ad[potassium] 5.2(hydrate)} (I):3 and 4L1H were mixed in 20\u2005ml of a 1M potassium acetate buffer solution. The mixture was left at 323\u2005K under stirring and nitro\u00adgen conditions for 1\u2005h. The mixture was then filtered and left to evaporate in air for six weeks, yielding yellow rod-like crystals of compound I (m.p. 553\u2005K decomposition).AgNO16H16Ag2N2O8S4, K2, 5.2(H2O), Mw = 880.175\u2005g\u2005mol\u22121: Calculated (%): C 21.88, H 2.99, N 3.18. Found (%): C 23.03, H 2.91, N 3.03. The small deviation is probably due to the loss of water mol\u00adecules of crystallization.Analysis for CESI\u2013MS: unstable under mass spectroscopy experimental conditions.\u22121) \u03bd: 3401(s), 2938(m), 1599(s), 1385(s), 1223(m).IR . It was not possible to locate the H atoms of the disordered water mol\u00adecules of crystallization. The residual electron density peaks of 1.14 and \u22121.10 e\u00c53 are at distances of 0.96 and 0.91\u2005\u00c5, respectively, from atom Ag1.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314622000773/im4015sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2414314622000773/im4015Isup2.hklStructure factors: contains datablock(s) I. DOI: 2143798CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N\u2010hydroxyalkyl phenylbenzoisoquinolindiones (PBIQs) has been developed. Incubation of plant material of Xiphidium caeruleum with hydroxylamines of various chain lengths resulted in 11 new 5\u2010hydroxy\u2010 and 5\u2010methoxy PBIQs with different N\u2010hydroxyalkyl side chain lengths. The antiproliferative effect and the cytotoxicity against HUVEC, K\u2010562, and HeLa cell lines of 26 previously reported PBIQs and the 11 newly synthesized N\u2010hydroxyalkyl PBIQs was determined for the first time. The results revealed that introducing long\u2010chain N\u2010aliphatic amine moieties improved the antiproliferative effect and cytotoxicity of PBIQs when compared to derivatives with N\u2010amino acids as side chains.A precursor\u2010directed approach to access Direct incubation ofXiphidium caeruleumplant material with \u03c9\u2010amino alcohols followed by extraction and purification results in N\u2010hydroxyalkyl phenylbenzoisoquinolindiones (PBIQs). Long\u2010chain derivatives show good antiproliferative effects together with low cytotoxicity. H\u2010benzo[de]isoquinoline\u20101,3(2H)\u2010diones,Xiphidium caeruleumLachnanthes tinctoriaWachendorfia thyrsiflora.Isoquinolines, which represent one of the largest groups of naturally occurring alkaloids, show a wide spectrum of biological activities.X. caeruleum were derived from a series of enzymatic hydrolyses and spontaneous conversions from three native phenylbenzoisochromenone (PBIC) glucosides, 1\u20133, and plant proteinogenic amino acids (see Supporting Information Table ST1).isoquinoline\u20101,6\u2010dione (31): Orange powder. HPLC\u2010PDA\u2010HRESIMS gradient 2, tR 7.5\u2005min; UV (MeCN\u2010H2O) \u03bbmax 206, 240, 320, 434\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST5; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 348.1237 [M+H]+ .2\u2010(4\u2032\u2032\u2010Hydroxybutyl)\u20105\u2010hydroxy\u20107\u2010phenyl\u20102H\u2010benzo[de]isoquinoline\u20101,6\u2010dione (32): Orange powder. HPLC\u2010PDA\u2010ESIMS gradient 2, tR 9.4\u2005min; UV (MeCN\u2010H2O) \u03bbmax 206, 238, 322, 442\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST5; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 362.1394 [M+H]+ .2\u2010(4\u2032\u2032\u2010Hydroxybutyl)\u20105\u2010methoxy\u20107\u2010phenyl\u20102H\u2010benzo[de]isoquinoline\u20101,6\u2010dione (33): Orange powder. HPLC\u2010PDA\u2010HRESIMS gradient 2, tR 8.4\u2005min; UV (MeCN\u2010H2O) \u03bbmax 196, 238, 320, 434\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST5; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 376.1541 [M+H]+ .2\u2010(6\u2032\u2032\u2010Hydroxyhexyl)\u20105\u2010hydroxy\u20107\u2010phenyl\u20102H\u2010benzo[de]isoquinoline\u20101,6\u2010dione (34): Orange powder. HPLC\u2010PDA\u2010HRESIMS gradient 2, tR 12.3\u2005min; UV (MeCN\u2010H2O) \u03bbmax 224, 324, 444\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST5; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 390.1708 [M+H]+ .2\u2010(6\u2032\u2032\u2010Hydroxyhexyl)\u20105\u2010methoxy\u20107\u2010phenyl\u20102H\u2010benzo[de]isoquinoline\u20101,6\u2010dione (35): Orange powder. HPLC\u2010PDA\u2010HRESIMS gradient 2, tR 10.9\u2005min; UV (MeCN\u2010H2O) \u03bbmax 206, 238, 320, 434\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST5; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 404.1863 [M+H]+ .2\u2010(8\u2032\u2032\u2010Hydroxyoctyl)\u20105\u2010hydroxy\u20107\u2010phenyl\u20102H\u2010benzo[de]isoquinoline\u20101,6\u2010dione (36): Orange powder. HPLC\u2010PDA\u2010HRESIMS gradient 2, tR 15.3\u2005min; UV (MeCN\u2010H2O) \u03bbmax 206, 218, 238, 324, 444\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST5; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 418.2030 [M+H]+ .2\u2010(8\u2032\u2032\u2010Hydroxyoctyl)\u20105\u2010methoxy\u20107\u2010phenyl\u20102H\u2010benzo[de]isoquinoline\u20101,6\u2010dione (37): Orange powder. HPLC\u2010DAD\u2010ESIMS gradient 2, tR 14.1\u2005min; UV (MeCN\u2010H2O) \u03bbmax 206, 238, 320, 434\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST6; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 432.2171 [M+H]+ .2\u2010(10\u2032\u2032\u2010Hydroxydecyl)\u20105\u2010hydroxy\u20107\u2010phenyl\u20102H\u2010benzo[de]isoquinoline\u20101,6\u2010dione (38): Orange powder. HPLC\u2010PDA\u2010HRESIMS gradient 2, tR 18.7\u2005min; UV (MeCN\u2010H2O) \u03bbmax 208, 234, 324, 444\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST6; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 446.2326 [M+H]+ .2\u2010(10\u2032\u2032\u2010Hydroxydecyl)\u20105\u2010methoxy\u20107\u2010phenyl\u20102H\u2010benzo[de]isoquinoline\u20101,6\u2010dione (39): Orange powder. HPLC\u2010DAD\u2010ESIMS gradient 2, tR 17.6\u2005min; UV (MeCN\u2010H2O) \u03bbmax 204, 238, 320, 434\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST6; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 460.2482 [M+H]+ .2\u2010(12\u2032\u2032\u2010Hydroxydodecyl)\u20105\u2010hydroxy\u20107\u2010phenyl\u20102H\u2010benzo[de]isoquinoline\u20101,6\u2010dione (40): Orange powder. HPLC\u2010DAD\u2010ESIMS gradient 2, tR 22.3\u2005min; UV (MeCN\u2010H2O) \u03bbmax 206, 242, 324, 444\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST6; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 474.2649 [M+H]+ .2\u2010(12\u2032\u2032\u2010Hydroxydodecyl)\u20105\u2010methoxy\u20107\u2010phenyl\u20102H\u2010benzo[de]isoquinoline\u20101,6\u2010dione (41): Orange powder. HPLC\u2010DAD\u2010ESIMS gradient 2, tR 21.2\u2005min; UV (MeCN\u2010H2O) \u03bbmax 204, 236, 320, 434\u2005nm; 1H\u2005NMR data, see Supporting Information Table ST6; 13C\u2005NMR data, see Supporting Information Table ST7; HRESIMS m/z 488.2806 [M+H]+ .30\u201341 were dissolved in dimethylsulfoxide . Five replicates were investigated for each assay. Cell toxicity assays were conducted using a HeLa cell line (DSM ACC 57), and antiproliferative activity was assayed using HUVEC (ATCC CRL\u20101730) and K\u2010562\u2005cell lines (DSM ACC 10). The test substances were dissolved in DMSO before being diluted in the respective cell culture medium to concentrations of between 1 and 100\u2005\u03bcg\u2009mL\u22121. The adherent cells were harvested in the logarithmic growth phase after soft trypsinization, using 0.25\u2009% trypsin in phosphate\u2010buffered saline containing 0.02\u2009%\u2009ethylenediaminetetraacetic acid. For each experiment, approximately 10,000 cells were seeded with 0.1\u2005mL culture medium per well of a 96\u2010well microplate. HeLa cells were pre\u2010incubated for 48\u2005h prior to the application of the test compounds to give subconfluent monolayers. Incubation was then conducted in a humidified atmosphere at 37\u2009\u00b0C and 5\u2009% CO2. In the case of K\u2010562 cells, the antiproliferative effect was determined using the CellTiter\u2010Blue1 assay.n HCl from the wells. The optical densities were measured at 660\u2005nm in a SUNRISE microplate reader .To determine cell toxicity and antiproliferative activity, PBIQs PLUS was performed as described earlier.Cell Death Detection ELISAAn overview of the compounds used for the bioassays together with their analytical data (MS and NMR) is provided in the Supporting Information. Furthermore, the results of the bioassays in graphical form and microscopic images of cells treated with PBIQs are given.The authors declare no conflict of interest.1As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "The chains are connected by C\u2014F\u22ef\u03c0(ring), C=O\u22ef\u03c0(ring) and slipped \u03c0-stacking inter\u00adactions. A Hirshfeld surface analysis of these inter\u00adactions was performed.The asymmetric unit consists of two independent mol\u00adecules differing in the orientation of the azido group. Each mol\u00adecule forms N\u2014H\u22efO hydrogen-bonded chains extending along the 8H7FN4O, consists of two independent mol\u00adecules differing in the orientation of the azido group. Each mol\u00adecule forms N\u2014H\u22efO hydrogen-bonded chains along along the c-axis direction with its symmetry-related counterparts and the chains are connected by C\u2014F\u22ef\u03c0(ring), C=O\u22ef\u03c0(ring) and slipped \u03c0-stacking inter\u00adactions. A Hirshfeld surface analysis of these inter\u00adactions was performed.The asymmetric unit of the title compound, C They have found valuable applications in medicinal chemistry \u2005\u00c5, C15\u22efCg1 = 3.902\u2005(2)\u2005\u00c5, C15=O2\u22efCg1 = 80.88\u2005(13)\u00b0] as shown in Fig.\u00a01p-tolyl analog x, \u2212y\u00a0+\u00a0z\u00a0+\u00a0A\u22efO2i hydrogen bonds form parallel chains for the second independent mol\u00adecule \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01; (iii) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a02] inter\u00adactions as well as by the C15=O2\u22efCg1 inter\u00adaction noted above and weak, slipped \u03c0-stacking between centrosymmetrically related C1\u2013C6 benzene rings , H\u22efH (13.5%), O\u22efH/H\u22efO (12.2%), C\u22efH/H\u22efC (11.9%) and F\u22efH/H\u22efF (9.7%), respectively. The two spikes in Fig.\u00a06d primarily represent the N\u2014H\u22efO hydrogen bonds but their breadth at longer values of di + de than at the tips indicate the contributions from C\u2014H\u22efO hydrogen bonds. Fig.\u00a07a showing all inter\u00admolecular inter\u00adactions and Fig.\u00a07b\u20137f those delineated into N\u22efH/H\u22efN (28.8%), H\u22efH (18.2%), C\u22efH/H\u22efC (12.6%), F\u22efH/H\u22efF (12.6%) and O\u22efH/H\u22efO (11.6%), respectively. Although the ordering of inter\u00adactions based on their percentage of the total is not the same as in the other mol\u00adecule, the percentages are not greatly different between the two and the corresponding plots are very similar type by type.A Hirshfeld surface analysis was performed with 6.N-(4-fluoro\u00adphen\u00adyl)acetamide (0.011\u2005mol), and sodium azide (0.015\u2005mol) were dissolved in a mixture of ethanol/water (70:30) and refluxed for 24\u2005h at 353\u2005K. After completion of the reaction , the 2-azido-N-(4-fluoro\u00adphen\u00adyl)acetamide that precipitated was filtered off and washed with cold water. A portion of the product was dissolved in hot ethanol, the solution was filtered, and the filtrate was left undisturbed for 7 days to form colorless, thick plate-like crystals.2-Chloro-\u22121) 3254 \u03c5 (N\u2014H amide), 1027 \u03c5 (N\u2014C amide), 1660 \u03c5 (C=O amide), 3073 \u03c5(C\u2014Harom), 1175 \u03c5(C\u2014N), 2961 \u03c5, 2109 \u03c5 (N3), 1H NMR (DMSO\u2013d6) \u03b4 ppm: 4.02 , 6.93\u20137.11 , 10.05 , 13C NMR (DMSO\u2013d6) \u03b4 ppm: 51.18 (CH2), 131.47 (Carom\u2014N), 113.90\u2013120.86 (Carom); 165.71 (C=O); HRMS (ESI\u2013MS) (m/z) calculated for C8H7FN4O 194.18; found 194.1165.Yield 69%, m.p. 358\u2013360K, FT\u2013IR global, I. DOI: 10.1107/S2056989022006764/vm2266Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022006764/vm2266Isup3.cmlSupporting information file. DOI: 2183049CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The ability of 4-[(benzyl\u00adamino)\u00adcarbon\u00adyl]-1-methyl\u00adpyridinium bromide salt to form hydrates was studied. Hirshfeld surfaces analysis was performed for identification of inter\u00admolecular inter\u00adactions. 14H15N2O+\u00b7Br\u2212\u00b70.5H2O, was studied by single-crystal and powder X-ray diffraction methods. In the asymmetric unit, two organic cations of similar conformation, two bromide anions and one water mol\u00adecule are present. In the crystal, N\u2014H\u22efBr hydrogen bonds link the cations and anions. The formation of a set of inter\u00admolecular C\u2014H\u22efBr and C\u2014H\u22ef\u03c0 inter\u00adactions result in double chains extending parallel to [011]. A Hirshfeld surface analysis showed high contributions of H\u22efH and C\u22efH/H\u22efC short contacts to the total Hirshfeld surfaces of the cations.The hemihydrate of 4-[(benzyl\u00adamino)\u00adcarbon\u00adyl]-1-methyl\u00adpyridinium bromide, C Polymorphic screening for this salt resulted in the crystallization of a hemihydrate. In this communication we present the mol\u00adecular and crystal structures of 4-[(benzyl\u00adamino)\u00adcarbon\u00adyl]-1-methyl\u00adpyridinium bromide hemihydrate, (C2.A and B), two bromide anions (A and B) and one water mol\u00adecule . The contribution of C\u22efH/H\u22efC short contacts is also significant . The Br\u22efH/H\u22efBr and O\u22efH/H\u22efO inter\u00adactions contribute to the total Hirshfeld surface in the same way .Inter\u00admolecular inter\u00adactions were analysed using Hirshfeld surface analysis and two-dimensional fingerprint plots by using rm Fig.\u00a04. The redrm Fig.\u00a04. The mairm Fig.\u00a04g,h. Thent Fig.\u00a05i,j. The5.et al., 2016et al., 2017sp3 and Csp3\u2014Car\u00adyl bonds.A search of the Cambridge Structural Database . A Rietveld refinement \u00adcarbon\u00adyl]-1-methyl\u00adpyridinium iodide , silver bromide and 700\u2005ml of water were loaded into a glass flask. The mixture was stirred for 72\u2005h, and the resulting precipitate was filtered off. The solvent was evaporated under reduced pressure. To the precipitate were added 300\u2005ml of aceto\u00adnitrile and refluxed for 2\u2005h. The reaction then was spontaneously cooled to a temperature of 303\u2005K and the precipitate filtered off and rinsed on the filter with 50\u2005ml of cooled aceto\u00adnitrile. The product was dried at 313\u2005K for 12\u2005h. Yield: 14\u2005g of 4-[(benz\u00adyl\u00adamino)\u00adcarbon\u00adyl]-1-methyl\u00adpyridinium bromide (28%); colourless crystals.8.Uiso(H) = 1.5Ueq for methyl groups and with Car\u2014H = 0.93\u2005\u00c5, Csp2\u2014H = 0.97\u2005\u00c5, Uiso(H) = 1.2Ueq for all other hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989022003784/wm5623sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022003784/wm5623Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022003784/wm5623Isup3.cmlSupporting information file. DOI: 2164796CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The \u2018tromancenium-8-carb\u00adoxy\u00adlic acid\u2019 entity with a hexa\u00adfluorido\u00adphosphate counter-ion represents a rare case of a cationic carb\u00adoxy\u00adlic acid. 7H7)(C6H5O2)]PF6 or [(Cht)Mn(Cp\u2019CO2H)]PF6, with Cht = cyclo\u00adhepta\u00adtrienyl and Cp\u2019 = C5H4, is an air-stable, purple, heteroleptic, cationic sandwich complex with manganese in oxidation state +I and \u03c0-coordinating cyclo\u00adhepta\u00adtrienyl and cyclo\u00adpenta\u00addienyl ligands. The latter ligand carries the carb\u00adoxy\u00adlic acid functionality. This \u2018tromancenium-8-carb\u00adoxy\u00adlic acid\u2019 with hexa\u00adfluorido\u00adphosphate as counter-ion represents a rare case of a cationic carb\u00adoxy\u00adlic acid. Structurally, this organometallic carb\u00adoxy\u00adlic acid displays the common motif of planar Osp2\u22efH\u2014Osp3/Osp3\u2014H\u22efOsp2 hydrogen-bonded carb\u00adoxy\u00adlic acid dimers with anti-oriented metallocenyl moieties, the cationic charge of which is balanced by octa\u00adhedrally shaped hexa\u00adfluorido\u00adphosphate anions. Positional disorder is observed in the cyclo\u00adhepta\u00adtrienyl ring and the PF6\u2212 anion.The title compound, [Mn(C As a result of the instability against nucleophiles of the parent compound tromancenium hexa\u00adfluorido\u00adphosphate \u2005\u00c5 is typical for a carbon\u2013carbon single bond. The C13\u2014O1 bond length of 1.205\u2005(10)\u2005\u00c5 is shorter than the C13\u2014O2 bond length of 1.303\u2005(10)\u2005\u00c5, which is coherent with the expectations.The mol\u00adecular entities of the title compound are shown in Fig.\u00a01y Table\u00a01, with tret al., 2014Cht/Cp bond lengths in the title compound. The C=O bond in cobaltocenium carb\u00adoxy\u00adlic acid is of the same length as the C\u2014O bond, due to disorder.The comparable organometallic compound cobaltocenium carb\u00adoxy\u00adlic acid hexa\u00adfluorido\u00adphosphate (Vanicek 6\u2212), which fill the space within the packing of the dimers were added. The solvents were removed on a rotary evaporator and the crude material dried in vacuo. The product was dissolved in aceto\u00adnitrile and filtered through a folded paper filter. Aceto\u00adnitrile was removed on a rotary evaporator and the product was dried in vacuo giving pure 8-tromancenium carb\u00adoxy\u00adlic acid hexa\u00adfluorido\u00adphosphate in 92% yield . Single crystals were obtained by diffusion crystallization in aceto\u00adnitrile out of diethyl ether at room temperature.A round-bottom flask was charged with 0.0563\u2005g of 8-carbo\u00admeth\u00adoxy\u00adtromancenium hexa\u00adfluorido\u00adphosphate \u03b4 = 4.89 , 5.21 , 6.93 ; signal of CO2H not observed due to rapid exchange. 13C NMR \u03b4 = 78.6 (ipso-carbon of Cp), 79.4 (C10/C11 of Cp), 80.3 (C9/C12 of Cp), 99.0 (C1\u20137 of Cht), 156.4 (CO2H). 55Mn NMR \u03b4 = 529. IR : 3000 (\u03bdO\u2014H + \u03bdC\u2014H), 1696 (\u03bdC=O), 1489, 1448, 1413, 1375 (\u03bdC\u2014OH + \u03bdC=C), 815 (\u03bdP\u2014F), 749 ), 600, 554 , 467, 437 (\u03bdMn). HRMS 255.0211 ([M\u00a0\u2212\u00a0PF6]+), calculated for C13H12O2Mn: 255.0212. UV\u2013vis \u03bbmax1 = 283, \u03bbmax2 = 559. Cyclic voltammetry (CV): \u0394E1/2 (Mn+/Mn2+) = 1.00\u2005V versus ferrocene/ferrocenium+ (irreversible).Properties: m.p.: 395.8\u2005K dec. d = 0.83\u2005\u00c5). A positional disorder in a ratio of 1:1 for the carbon atoms and attached hydrogen atoms of the seven-membered ring: C1\u2013C7: C1A\u2013C7A was considered; the corresponding carbon atoms were refined with isotropic displacement parameters. A further positional disorder of all fluorine atoms of the PF6\u2212 anion was refined in ratio 45:55 for F1\u2013F6:F1A\u2014F6A with anisotropic displacement parameters. In an alternative model, the crystal structure was also refined in the non-centrosymmetric space group P1 with a new data set, for which TWINABS in the P1 model and several remaining electron-density peaks between the carbon atoms of the two seven-membered rings clearly show that the disorder will be retained in the non-centrosymmetric space group. Hence, the latter was discarded and the centrosymmetric model used for final processing.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314623001074/wm4180sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314623001074/wm4180Isup2.hklStructure factors: contains datablock(s) I. DOI: 2239883CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Correction: \u201cAbsolute quantitation of microbes using 16S rRNA gene metabarcoding: A rapid normalization of relative abundances by quantitative PCR targeting a 16S rRNA gene spike\u2010in standard\u201d by Olivier Zemb | Caroline S. Achard | Jerome Hamelin | Marie\u2010L\u00e9a De Almeida | B\u00e9atrice Gabinaud | Laurent Cauquil | Lisanne M.G. Verschuren | Jean\u2010Jacques Godon published online on 11 January\u00a0Figure 1 of the original version of published article has been replaced. The correct figure is placed below: In the original version, in the text (section 2.3) \u201cforward and reverse primers E 5\u2032\u2010CAGATGTGAAATCATCGATCG/5\u2032\u2010CCGATTTCAATCGTACACCTG\u201d should read \u201cforward and reverse primers E 5\u2032\u2010ACGTAGAATTCCAGGTGTACGAT/5\u2032\u2010ACCTGAGCGTCAGTCTTCGT\u201d."} +{"text": "Scientific Reports 10.1038/s41598-021-82599-1, published online 04 February 2021Correction to: The original version of this Article contained an error in the Methods section, where \u201c7.8\u2009\u00d7\u2009300\u00a0mm\u201d was incorrectly given as \u201c4.6\u2009\u00d7\u2009300\u00a0mm\u201d.In the Methods section, under the subheading \u2018Size-exclusion chromatography\u2019,\u201cA SEPAX SRT SEC-1000 column (4.6\u2009\u00d7\u2009300\u00a0mm) and guard column were used for all SEC-MALS experiments.\u201dnow reads:\u201cA SEPAX SRT SEC-1000 column (7.8 \u00d7 300 mm) and guard column were used for all SEC-MALS experiments.\u201dThe original Article has been corrected."} +{"text": "The a new polymorph of the title compound is reported in which the C\u2014H\u22efO hydrogen bonds and \u03c0-\u03c0 stacking inter\u00adactions link mol\u00adecules into the layers in the crystal. 14H8Br3N3O2, (form-2) was obtained in the same manner as the previously reported form-1 . The structure of the new polymorph is stabilized by a C\u2014H\u22efO hydrogen bond that links mol\u00adecules into chains. These chains are linked by face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions, resulting in a layered structure. Short inter-mol\u00adecular Br\u22efO contacts and van der Waals inter\u00adactions between the layers aid in the cohesion of the crystal packing. In the previously reported form-1, C\u2014H\u22efBr inter\u00adactions connect mol\u00adecules into zigzag chains, which are linked by C\u2014Br\u22ef\u03c0 inter\u00adactions into layers, whereas the van der Waals inter\u00adactions between the layers stabilize the crystal packing of form-2. Hirshfeld mol\u00adecular surface analysis was used to compare the inter\u00admolecular inter\u00adactions of the polymorphs.A new polymorph of the title compound, C al. 2022. Acta Cr Crystal Explorer 17.5 to +1.1715 (blue) a.u. , H\u22efH (12.8%), C\u22efH/H\u22efC (11.5%) and O\u22efH/H\u22efO (10.6%) contacts are shown in Fig.\u00a06b\u2013e, while the numerical details for the shortest contacts are given in Table\u00a02viz. Br\u22efH/H\u22efBr, H\u22efH, C\u22efH/H\u22efC and O\u22efH/H\u22efO contacts -1--2-(4-bromo\u00adphen\u00adyl)diazene unit showed that the ten closest are those of CSD refcodes HEHKEO (I) , resulting in zigzag C(8) chains along the [100] direction. These chains are connected by C\u2014Br\u22ef\u03c0 inter\u00adactions into layers parallel to (001). van der Waals inter\u00adactions between the layers contribute to the crystal cohesion.C\u2014H\u22efBr inter\u00adactions connect the mol\u00adecules in the crystal of the form-1 polymorph of the title compound, (II) are joined into layers parallel to (011) by C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds. C\u2014Br\u22ef\u03c0 and C\u2014F\u22ef\u03c0 contacts, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions, strengthen the crystal packingThe mol\u00adecules in (III) are connected into chains running parallel to [001] by C\u2014H\u22efO hydrogen bonds. C\u2014F\u22ef\u03c0 contacts and \u03c0\u2013\u03c0 stacking inter\u00adactions help to consolidate the crystal packing, and short Br\u22efO [2.9828\u2005(13)\u2005\u00c5] distances are also observed.The mol\u00adecules in the crystal of (IV), the mol\u00adecules are linked into inversion dimers via short halogen\u2013halogen contacts . No other directional contacts could be identified, and the shortest aromatic ring-centroid separation is greater than 5.25\u2005\u00c5.In the crystal of (V) and (VI), the mol\u00adecules are linked through weak X\u22efCl contacts [X = Cl for (V) and Br for (VI)], C\u2014H\u22efCl and C\u2014Cl\u22ef\u03c0 inter\u00adactions into sheets lying parallel to (001).In the crystals of (VII), the mol\u00adecules are stacked in columns along [100] via weak C\u2014H\u22efCl hydrogen bonds and face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions. The crystal packing is further consolidated by short Cl\u22efCl contacts.In the crystal of (VIII), mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds into zigzag chains running parallel to [001]. The crystal packing also features C\u2014Cl\u22ef\u03c0, C\u2014F\u22ef\u03c0 and N\u2014O\u22ef\u03c0 inter\u00adactions.In (IX), C\u2014H\u22efN and short Cl\u22efCl contacts are observed, and in (X), C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and short Cl\u22efO contacts occur.In -1-(4-bromo\u00adphen\u00adyl)-2-(3-nitro\u00adbenzyl\u00adidene)hydrazine (1\u2005mmol), tetra\u00admethyl\u00adethylenedi\u00adamine , CuCl and CBr4 (4.5\u2005mmol). After 1\u20133\u2005h , the reaction mixture was poured into a 0.01 M solution of HCl , and extracted with di\u00adchloro\u00admethane (3 \u00d7 20\u2005mL). The combined organic phase was washed with water (3 \u00d7 50\u2005mL), brine (30\u2005mL), dried over anhydrous Na2SO4 and concentrated in vacuo using a rotary evaporator. The residue was purified by column chromatography on silica gel using appropriate mixtures of hexane and di\u00adchloro\u00admethane (3/1\u20131/1). Crystals suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution. Red solid (62%); m.p. 391\u2005K. Analysis calculated for C14H8Br3N3O2 (M = 489.95): C 34.32, H 1.65, N 8.58; found: C 34.29, H 1.66, N 8.55%. 1H NMR \u03b4 7.90\u20137.44 . 13C NMR \u03b4 150.88, 148.57, 148.12, 132.81, 132.47, 132.25, 130.04, 126.40, 125.30, 124.53, 123.57, 94.10. ESI-MS: m/z: 490.91 [M + H]+.This dye was synthesized according to the reported method (Akkurt 6.Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022007113/yk2172sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022007113/yk2172Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022007113/yk2172Isup3.cmlSupporting information file. DOI: 2189348CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Over the past 20\u00a0years, the interest in the biological chemistry of HNO has increased significantly due to the numerous beneficial pharmacological effects of its donors. Increased availability of various HNO donors was accompanied by great progress in the understanding of HNO chemistry and chemical biology. This review is focused on the chemistry of HNO, with emphasis on reaction kinetics and mechanisms in aqueous solutions.Azanone is the protonated form of the product of one-electron reduction of nitric oxide ( Formallificance . Interesificance . This resulfide HS and azaazanone\u201d or \u201chydridooxidonitrogen\u201d (which name is based on additive nomenclature). The commonly used name is \u201cnitroxyl.\u201d It has also been used to describe azanone anion, NO\u2212 [which IUPAC recommended name is \u201coxidonitrate (1\u2212)\u201d]. The term \u201cnitroxyl\u201d may be misleading as aminoxyl radicals, of the general formula R2N-O\u2022, are known as \u201cnitroxyl radicals.\u201d Therefore, the reader must be aware that a literature search with the keyword \u201cnitroxyl\u201d may yield references to aminoxyl radicals R2N-O\u2022. In some of the scientific literature names \u201cnitroso hydrogen,\u201d \u201cmonomeric hyponitrous acid,\u201d \u201cnitrosyl hydride\u201d or \u201chydrogen oxonitrate,\u201d were also used. For the compliance with the IUPAC recommendations and to avoid ambiguity, we suggest the use of the name \u201cazanone\u201d for HNO and \u201cazanone anion\u201d for NO\u2212.Prior to describing HNO chemistry, it is important to briefly discuss the recommended and the used nomenclature. The IUPAC recommended name of HNO is \u201cN-O, the N-H bond length rN-H and the H-N-O angle \u03b8H-N-O. The qualitative discussion of the electronic structure of HNO has been given by N-O = 1.212\u00a0\u00c5, rN-H = 1.063\u00a0\u00c5, and \u03b8H-N-O = 108.6\u00b0. HNO structure and NO bond vibrational frequency \u03c5NO has been the subject of numerous theoretical studies . Deprot\u2212 a pKa value of 4.7 for HNO was reported in 1970 (Ka(1HNO/3NO\u2212) value of 11.4 is recommended. Shafirovich and Lymar have also estimated the pKa values for other HNO acid-base equilibria: pKa(1HNO/1NO\u2212) \u223c 23, pKa(3HNO/3NO\u2212) = \u22121.8 = \u22121.8 , suggest\u2022NO, due to its electronic structure, has very low value \u2014 Eea = 0.026 \u00b1 0.005\u00a0eV = \u22120.81\u00a0V, and practically outside of the biologically compatible range (\u2022NO reduction leads to the formation of HNO. Taking into account the postulated pKa(1HNO/3NO\u2212) value of 11.4, Lymar and co-workers, estimated standard redox potential for \u2022NO/HNO redox pair to be equal to \u22120.14\u00a0V (E\u00b0(\u2022NO/HNO) = \u22120.14\u00a0V, and E\u00b0\u2019(\u2022NO/HNO) = \u22120.55\u00a0V for pH 7) (E\u00b0(\u2022NO/HNO) = 0.27\u00a0V, and E\u00b0\u2019(\u2022NO/HNO) = \u22120.14\u00a0V for pH 7, based on the computational reevaluation of HNO solvation free energy (2/O2\u2022\u2013 redox couple), would suggest the feasibility of \u2022NO reduction to HNO by a variety of cellular enzymatic redox centers. The \u2022NO/HNO interconversion under physiological conditions has been recently discussed in detail (Electron affinity of 0.005\u00a0eV . \u2022NO redle range . At physor pH 7) . Those ve energy . Such van detail , and is 2N2O2 (k1a = (8 \u00b1 3) \u00d7 106\u00a0M\u22121s\u22121) (2N2O2 have been reported with pKa (H2N2O2/HN2O2\u2212) = 7.0 and pKa (HN2O2\u2212/N2O22\u2013) = 10.9. Therefore, at physiological pH (pH > 7), H2N2O2 exists predominantly in its mono-deprotonated form (HN2O2\u2212). This form is responsible for the instability of H2N2O2 in aqueous solutions and for the formation of N2O .It should be noted, that direct experimental determination of the chemical reactivity of HNO is hampered by its rapid dimerization to hyponitrous acid H\u00a0M\u22121s\u22121) . This rain situ, e.g., with the use of radiation techniques involving one-electron reduction of \u2022NO by solvated electrons or by hydrogen atoms (H\u2022), or photochemical techniques , including photolysis of alkaline aqueous solutions of trioxodinitrate . Below, the radiation chemistry of aqueous solutions of \u2022NO and the laser flash photolysis of aqueous solutions of N2O32\u2212 will be described, as those methods greatly contributed to our understanding of azanone chemistry and to the determination of the rate constants of the reactions involved. Further development of such methodologies for rapid in situ formation of HNO/NO\u2212and for monitoring their reactions may overcome current obstacles in the studies of HNO/NO\u2212chemistry and result in improved knowledge of the azanone chemistry and its biological fates.Due to the instability of HNO in aqueous solutions, in chemical studies on HNO reactivity it is necessary to use HNO donors, that release HNO in a controlled manner, or produce azanone \u2022NO leads to the formation of nitrite (NO2\u2212) and N2O. The primary transient reactive species produced within approximately 10\u20138\u00a0s after the exposition of water to ionizing radiation are hydroxyl radical (\u2022OH), hydrated electron (eaq\u2212), and hydrogen atom (H\u2022) [ONOO\u2212]where kvi in the presence of azanone scavenger is expressed by 2 and the scavenger studied.\u2212, or such reaction is outcompeted by the reaction between ONOO\u2212and the boronate probe; 3) the scavenger does not react with or interferes with the detection of the product of oxidation of the boronic probe; and 4) the amount of HNO released is significantly lower than the amount of the scavenger and of O2 in the solution.The rate of ArOH formation r[donor]Comparis2\u2212 was determined to be equal to (5.0 \u00b1 0.9) \u00d7 103\u00a0M\u22121s\u22121 results in the formation of molecular nitrogen (N2) and water (k23 = (4.0 \u00b1 0.3) \u00d7 103\u00a0M\u22121s\u22121. Recent analysis of the competition between O2 and NH2OH for HNO using the boronate reporter for ONOO\u2212, resulted in the rate constant equal to (2.1 \u00b1 0.4) \u00d7 104\u00a0M\u22121s\u22121 \u00d7 106\u00a0M\u22121s\u22121, k25 = (1.2 \u00b1 0.2) \u00d7 106\u00a0M\u22121s\u22121 and k26 = (2.2 \u00b1 0.7) \u00d7 104\u00a0M\u22121s\u22121, for HS\u2212, HSO3\u2212, and S2O32\u2212, respectively (2O32\u2212 (4\u00a0M\u22121s\u22121 (\u2212 and HNO (\u2212 to form HS2\u2212 and release NH2OH (\u2212O3SSNHOH adduct (the product of 2O32\u2212 could result in the formation of NH2OH and S4O62\u2212 anion (2OH (2S shows that this reaction leads to the formation of hydrogen polysulfides (H2Sn) or sulfur S8, depending on the relative concentrations of H2S and HNO. The reactivity of HNO towards selected inorganic scavengers is summarized in The second-order rate constants were determined to be equal to: ectively . The rat and the glutathione disulfide NH2 are formed from a single transient intermediate, supporting the hypothesis of GSNHOH formation (2 is not observed with other reactive nitrogen species (RNS): \u2022NO, nitrogen dioxide (\u2022NO2), ONOO\u2212, or N2O3, making GS(O)NH2 a unique and HNO-specific product. GS(O)NH2 was, therefore, used as a marker for HNO generation from several biologically-relevant pathways: thiol-mediated decomposition of S-nitrosothiols, peroxidase-dependent NH2OH and N-hydroxy-L-arginine oxidation (Exposure of reduced glutathione (GSH) to HNO was shown to lead to the formation of both the sulfinamide derivative (GS(O)NHe, GSSG) . Higher ormation . It has xidation .It has been long assumed that HNO-driven oxidative transformation of thiols to sulfinamides is irreversible. C-terminal cysteines the nature of HNO-derived modifications is highly affected by the C-terminal carboxylate .2\u2212 and O2, it was concluded that the rate constant for the reaction of GSH with NO\u2212/HNO is significantly higher than 1 \u00d7 105\u00a0M\u22121s\u22121 (N-acetyl cysteine (NAC) were determined to be equal to 2 \u00d7 106\u00a0M\u22121s\u22121 and 5 \u00d7 105\u00a0M\u22121s\u22121, respectively (6\u00a0M\u22121s\u22121 (2 for HNO (using boronate probes for ONOO\u2212), yielded the following values of the rate constants in aqueous solutions at pH 7.4: cysteine (kCys = (4.5 \u00b1 0.9) \u00d7 106\u00a0M\u22121s\u22121, pKa = 8.3), glutathione (kGSH = (3.1 \u00b1 0.6) \u00d7 106\u00a0M\u22121s\u22121, pKa = 8.8), N-acetylcysteine (kNAC = (1.4 \u00b1 0.3) \u00d7 106\u00a0M\u22121s\u22121, pKa = 9.5) and captopril (kCap = (6 \u00b1 1) \u00d7 105\u00a0M\u22121s\u22121, pKa = 9.8) is the active form reacting with HNO. The second order rate constants for HNO reaction with protein-bound thiols were also determined in that study for human and bovine serum albumins (pKa \u223c 8.5 for Cys34) (kHSA = (1.4 \u00b1 0.4) \u00d7 106\u00a0M\u22121s\u22121, kBSA = (1.4 \u00b1 0.3) \u00d7 106\u00a0M\u22121s\u22121). Such a high reactivity of thiols towards HNO, together with high concentrations of GSH and albumin in cells and plasma, respectively, suggest that thiols may be the potential targets of HNO in biological systems.Several groups studied the kinetics of the reaction of HNO with GSH. Based on the effect of GSH on the rate of Angeli\u2019s salt decomposition in aqueous solution at pH 7.4 in the presence of NO5\u00a0M\u22121s\u22121 . In an iectively . The rat6\u00a0M\u22121s\u22121 . More rea = 9.8) . It was N-acetylcysteine, bovine serum albumin and human serum albumin was also recently reported (kobs) can be expressed as a function of pH, which depends on thiol pKa, and the rate constants of azanone reaction with RS\u2212 (kthiolate) and with RSH (kthiol) (\u2212) is favored (kthiolate \u223c 107\u00a0M\u22121s\u22121) over reactions with the thiol . Such observation is consistent with the observed inverse correlation between the reaction rate constant and the pKa value of the thiol.The pH-dependent kinetics of HNO reactions with selected thiols\u2013glutathione, reported . It was kthiol) :kobs=ktKa and inc species .6H5SO2\u2212), the decomposition product of Piloty\u2019s acid, has been determined as kbenzenesulfinate = (4.4 \u00b1 0.9) \u00d7 104\u00a0M\u22121s\u22121 would be expected to be the products of those reactions . The rat4\u00a0M\u22121s\u22121 .RSO2-+H2\u2212, 2-ClPhSO2\u2212 and 2-CF3PhSO2\u2212) were also determined and are equal to (5.0 \u00b1 1.2) \u00d7 104\u00a0M\u22121s\u22121, (3.0 \u00b1 0.7) \u00d7 104\u00a0M\u22121s\u22121 and (1.1 \u00b1 0.2) \u00d7 104\u00a0M\u22121s\u22121, respectively (3\u00a0M\u22121s\u22121), the rate constant for the reaction between HNO and benzenesulfinate is one order of magnitude higher.The rate constants of HNO reactions with 2-bromo-, 2-chloro- and 2-trifluoromethylbenzenesulfinates was reported with reported . It was S-diazeniumdiolate transient product has also been recently proposed for the reaction of thiols with \u2022NO, resulting in the formation of HNO \u00d7 104\u00a0M\u22121s\u22121 .C- and S-nitroso compounds resulting in the formation of phosphine oxides and aza-ylides (kGSH = 2 \u00d7 106\u00a0M\u22121s\u22121), the rate constant for the reaction of HNO with a water-soluble phosphine, trisphosphine trisodium salt, was estimated at 37\u00b0C and pH 7.4 to be equal to 9 \u00d7 105\u00a0M\u22121s\u22121 \u00d7 107\u00a0M\u22121s\u22121 were recently re-evaluated and the values determined at pH 7.4 and 25\u00b0C are equal to (3.0 \u00b1 0.5) \u00d7 107\u00a0M\u22121s\u22121 . The val6\u00a0M\u22121s\u22121 .5\u00a0M\u22121s\u22121.In 2015 C-nucleophiles can act as efficient scavengers of HNO , 1,3-cyclohexanedione (C6H8O2), and 1,3-cycloheptanedione (C7H10O2) was studied and the obtained second order rate constants were equal to 2.8 \u00d7 102\u00a0M\u22121s\u22121, 2.2 \u00d7 103\u00a0M\u22121s\u22121 and 6.8 \u00d7 103\u00a0M\u22121s\u22121, respectively. This indicates that the reactivity of cyclic C-nucleophiles toward HNO significantly increases with an increase in their ring size -2,4-piperidinedione = 1.4 \u00d7 104\u00a0M\u22121s\u22121.The substitution of one or more carbon atoms in the ring of 1,3-cyclohexanedione with nitrogen resulted in an increase in ward HNO . In the k2-thiobarbituric acid = 8.2 \u00d7 102\u00a0M\u22121s\u22121 and kMeldrum\u2019s acid = 8.7 \u00d7 102\u00a0M\u22121s\u22121.The reactivity of barbituric acid, 1,3-dimethylbarbituric acid, 2-thiobarbituric acid and Meldrum\u2019s acid towards azanone was alsoThe reactivity of HNO towards different classes of organic nucleophiles has been summarized in \u2022NO (E\u00b0\u2019 (\u2022NO/1HNO) = \u22120.16\u00a0V for pH 7) ) , it coul 0.256\u00a0V ), Cu,Zn- 0.403\u00a0V ), aminox 0.204\u00a0V ), TEMPOL 0.204\u00a0V ) and 3-c 0.102\u00a0V ) and nit 0.270\u00a0V ) and c-P 0.270\u00a0V ).c(Fe3+) with an excess of Angeli\u2019s salt in phosphate buffer at pH 7.0 at 25\u00b0C leads to the formation of ferrocytochrome c (cyt c(Fe2+)) (c(Fe3+) reduction was nearly zero order in the concentration of cyt c(Fe3+) and first order in the concentration of HN2O3\u2212, suggesting that the decomposition of Angeli\u2019s salt was controlling the rate of the reaction, consistent with HNO acting as the reductant of cyt c (Fe3+) (c reduction at low [HN2O3\u2212]0/[cyt c (Fe3+)]0 ratio was close to 50%.In 1988 it was reported that the treatment of cyt c(Fe2+)) . The inic (Fe3+) . The repc(Fe3+) reduction by HNO have been reported in the literature: k39 = 2 \u00d7 104\u00a0M\u22121s\u22121, (k39 = 4 \u00d7 104\u00a0M\u22121s\u22121 (c(Fe3+) in aqueous solutions of Angeli\u2019s salt was not observed in the presence of O2 (c (Fe2+) is oxidized to cyt c(Fe3+) in aerated solutions of Angeli\u2019s salt (\u2212 in the reaction of HNO and O2 (c(Fe2+) by ONOO\u2212 or ONOO authors .cyt\u00a0c. In an elegant kinetic study on the reactivity of nitroxides toward HNO with the use of EPR spectrometry, the reduction of cyclic stable aminoxyl radicals (RN-O\u2022) by HNO to the corresponding hydroxylamines (RN-OH) (Other mild one-electron oxidants reported to oxidize HNO to (RN-OH) was repo (RN-OH) .RN-O\u2022\u00a0+\u2022 radicals have been determined with the use of the competition kinetics against the reference reaction of HNO with ferric myoglobin (MbFeIII). In the presence of HNO donor, MbFeIII undergoes reductive nitrosylation with the formation of relatively stable ferrous nitrosyl complex, MbFeIINO, which can be followed by UV-Vis absorption spectroscopy. Using kMbFe(III) = 8 \u00d7 105\u00a0M\u22121s\u22121 (kTEMPO = (1.4 \u00b1 0.2) \u00d7 105\u00a0M\u22121s\u22121), TEMPOL (kTEMPOL = (1.4 \u00b1 0.2) \u00d7 105\u00a0M\u22121s\u22121) and 3-carbamoyl-PROXYL (k3-CP = (4.3 \u00b1 0.4) \u00d7 104\u00a0M\u22121s\u22121) (kTEMPOL = 8 \u00d7 104\u00a0M\u22121s\u22121) (kTEMPO = 6.3 \u00d7 104\u00a0M\u22121s\u22121) (k = (8 \u00b1 2) \u00d7 104\u00a0M\u22121s\u22121) \u00a0\u00d7\u00a0104\u00a0M\u22121s\u22121) (The second-order rate constants of HNO reactions with RN-O5\u00a0M\u22121s\u22121 as a ref\u00a0M\u22121s\u22121) . Those v\u00a0M\u22121s\u22121) and TEMP\u00a0M\u22121s\u22121) and a mo\u00a0M\u22121s\u22121) . Nitroxi\u00a0M\u22121s\u22121) . This pr\u00a0M\u22121s\u22121) . The pro4\u00a0M\u22121s\u22121 , a value\u00a0M\u22121s\u22121) . Due to \u2022NO will react with HNO to form the N2O2\u2022\u2212 radical anion , are widely used in redox biology studies as scavengers of \u2022NO of characteristic EPR spectrum and \u2022NO2 (\u2022NO are in the range (0.4\u201316) \u00d7 104\u00a0M\u22121s\u22121 = 8 \u00d7 105\u00a0M\u22121s\u22121 (kPTIO = kc-PTIO = (1.4 \u00b1 0.2) \u00d7 105\u00a0M\u22121s\u22121. Independently, the kinetics of c-PTIO reaction with HNO was determined using NH2OH as a competing agent and the kC-PTIO/k23 ratio of 5.5 \u00b1 0.9 was obtained (k23 = (4.0 \u00b1 0.3) \u00d7 103\u00a0M\u22121s\u22121 (k23 = (2.1 \u00b1 0.4) \u00d7 104\u00a0M\u22121s\u22121 (kC-PTIO = (2.2 \u00b1 0.5) \u00d7 104\u00a0M\u22121s\u22121 (kC-PTIO = (1.2 \u00b1 0.4) \u00d7 105\u00a0M\u22121s\u22121. It has been also shown, that \u2022NO2 radicals formed in such a system can oxidize Angeli\u2019s salt anion to transient radical product, that decomposes to NO2\u2212 and \u2022NO (k43 = 7.7 \u00d7 106\u00a0M\u22121s\u22121) .N2O32-+64\u2212 anion as a \u2022NO2 radical scavenger (64\u2212 and Fe(CN)63\u2212 anions are in redox equilibrium with nitronyl nitroxide radicals and Fe(CN)63\u2212 anion is an oxidant of HNO. Based on the established competition, the kc-PTIO/k23\u00a0ratio of 1.7 \u00b1 0.2 and k23 = (4.0 \u00b1 0.3) \u00d7 103\u00a0M\u22121s\u22121 (k23 = (2.1 \u00b1 0.4) \u00d7 104\u00a0M\u22121s\u22121 (kc-PTIO = (6.8 \u00b1 1.3) \u00d7 103\u00a0M\u22121s\u22121 and 2\u00a0M\u22121s\u22121 (kC-PTIO = (3.6 \u00b1 1.1) \u00d7 104\u00a0M\u22121s\u22121 and 3\u00a0M\u22121s\u22121 cavenger . Such a 3\u00a0M\u22121s\u22121 or k23 4\u00a0M\u22121s\u22121 one can 2\u00a0M\u22121s\u22121 or kC-P3\u00a0M\u22121s\u22121 .c (cyt c), soluble guanylate cyclase (sGC), and cytochromes P450 (CYP450) (II)O2) reacts with HNO forming ferric heme and possibly \u2022NO and H2O2 (k44 = 1 \u00d7 107\u00a0M\u22121s\u22121 (\u2022NO formed upon HNO oxidation reacts with another Mb (FeII)O2 molecule with rate constant k45 = 3\u20135 \u00d7 107\u00a0M\u22121s\u22121 producing ferric heme and NO3\u2212 (II)O2 (II)O2 and Hb(FeII)O2 convert HNO to \u2022NO rapidly. It is worth to emphasize that in the literature an alternative mechanism for the heme (FeII)O2/HNO reaction was also proposed. In that mechanism, two ferric hemes, hydroxylamine and O2 are the final products (The reaction of HNO with heme-proteins has been studied extensively for nearly 40\u00a0years. Among the studied heme-enzymes are myoglobin (Mb), hemoglobin (Hb), horseradish peroxidase (HRP), catalase (cat), cytochrome (CYP450) . The reaand H2O2 (Doyle aand H2O2 and the 7\u00a0M\u22121s\u22121 . \u2022NO fornd NO3\u2212 (Doyle and NO3\u2212 . Identic\u2212 (II)O2 . Overallproducts .heme-FeII)) reacts with HNO forming coordination complex Mb (FeII)HNO (4\u00a0M\u22121s\u22121 (5\u00a0M\u22121s\u22121 (II) with HNO is followed by subsequent reaction of Mb (FeII)HNO with HNO yielding Mb (FeIII)NO, with the rate constant estimated as 1.67 \u00d7 104\u00a0M\u22121s\u22121 (II) is equal to ca. 2.0 \u00d7 105\u00a0M\u22121s\u22121 (5\u00a0M\u22121s\u22121 (Deoxymyoglobin (Mb (FeFeII)HNO . Origina4\u00a0M\u22121s\u22121 . The re-5\u00a0M\u22121s\u22121 . The rea4\u00a0M\u22121s\u22121 (Zapata 4\u00a0M\u22121s\u22121 . For deo5\u00a0M\u22121s\u22121 . For oth5\u00a0M\u22121s\u22121 .heme-FeII)NO and Hb(FeII)NO, respectively .heme-FeThe rate constant of reductive nitrosylation of metHb is difficult to determine due to the side reaction of HNO with \u03b2-93 cysteines of hemoglobin . Nonethe5\u00a0M\u22121s\u22121 and 2 \u00d7 106\u00a0M\u22121s\u22121 with HNO results in the reduction of iron heme and the formation of \u2022NO, as already discussed in Oxidation of HNO to NO) (The reaction of cyt O to NO) . It has O to NO) .\u2022NO (II)HNO complex (II)NO. sGC(FeIII) is unreactive toward HNO or \u2022NO. However, a contrary report was published, where the HNO-dependent activation of sGC was excluded (II)HNO and HNO that follows to the formation of sGC(FeII)NO (In the case of sGC there is no estimated rate constant for its reaction with HNO. Miller and coworkers proposed that activation of sGC proceeds directly through HNO and the ferrous heme reaction without conversion of HNO to \u2022NO . The exp complex but not excluded . That re(FeII)NO is respo(FeII)NO .II(NO) complex similarly to other heme enzymes .4\u00a0M\u22121s\u22121) is in re\u00a0M\u22121s\u22121) indicatiThe kinetics of HNO reaction with other porphyrins containing iron and manganese were also studied and the appropriate rate constants are compiled in via the possible metal porphyrin-donor complex ) is an important example, since it occurs naturally and is an essential mammalian coenzyme. It was shown that aquacobalamin (H2O-Cbl+) and hydroxycobalamin (HO-Cbl) in the presence of Angeli\u2019s salt are converted to the nitroxyl cobalamin (NO\u2212-CoIIICbl) (2O-Cbl+ occurs and the complex was observed. The rate constant for the AS/H2O-Cbl+ reaction at acidic pH was determined to be 122.6 \u00b1 5.3 M\u22121s\u22121 (2O-Cbl+ and Piloty\u2019s acid (PA) has shown that PA also reacts directly with H2O-Cbl+ at neutral pH (28.4 \u00b1 0.4\u00a0M\u22121s\u22121) (\u2212-CoIIICbl) . The mec3 M\u22121s\u22121 . Recent \u00a0M\u22121s\u22121) . In turn\u00a0M\u22121s\u22121) . The mec\u00a0M\u22121s\u22121) ; 2016. ToIIICbl) ; 2016.To summarize, the reaction scheme of HNO with heme-proteins is very complex and still remains an area that requires further studies and analyses. In addition, an interesting aspect relates to the direct reactions of HNO-donors with metal-porphyrins, of potential pharmacological relevance.2O3\u2212, however the exact stoichiometry of this reaction with respect to HNO is unclear (c(Fe3+) (10\u201340\u00a0\u03bcM), the Cu,Zn-SOD-inhibitable reduction of cyt c(Fe3+) was observed. The Cu,Zn-SOD concentration was selected to be sufficient to compete with cyt c(Fe3+) for O2\u2022\u2013, but too low to detectably inhibit NADPH oxidation. That experiment strongly suggested the generation of O2\u2022\u2013 in HNO/O2/NADPH system. The authors proposed that NADPH is oxidized in such a solution to NADP\u2022 radicals, which subsequently reduce O2 to O2\u2022\u2212 with the formation of NADP+. At higher concentration (\u223c2\u00a0\u03bcM), Cu,Zn-SOD inhibited by \u223c50% the oxidation of NADPH \u00d7 104\u00a0M\u22121s\u22121 was reported (k52 = (1.3 \u00b1 0.4) \u00d7 104\u00a0M\u22121s\u22121 was also determined for NADPH. In the direct reaction of HNO with NADH or NADPH in deaerated Angeli\u2019s salt solution, one may expect the formation of hydroxylamine as an HNO reduction product with Angeli\u2019s salt in aerobic buffered aqueous solutions (pH 7.4) has been presented in several scientific reports (N-nitroso derivative as the main product. The yield of its formation is rather low (below 20%) and depends on the indolic compound: Angeli\u2019s salt concentrations ratio (N-nitroso derivative is significantly higher in the presence of bicarbonate (N-nitrosation mechanism. The recent study of N-acetyl-L-tryptophan and small peptides containing either tryptophan or both tryptophan and cysteine residues, has shown that in the presence of cysteine, excess HNO is required for efficient TrpNO formation, clearly indicating, that tryptophan residues are significantly less reactive towards HNO, than cysteine residues.The results of the studies on the reaction of various indolic compounds (e.g., reports . All thens ratio . It was arbonate , which m\u2022NO, and may be expected to exhibit unique physiological activity. The experimental limitations due to its instability under physiological conditions provide a significant hindrance in study design to further explore HNO chemistry. The development of new donors and rapid generation methods for time-resolved kinetic studies may overcome some of those obstacles. In addition, the combination of the experimental data with theoretical calculations may help with drawing reliable mechanistic conclusions. Finally, as a note of caution, it has to be emphasized that many rate constants of HNO reactions listed in this review were determined by competition kinetics, via kinetic simulations or other indirect approaches, and are based on very few values determined directly. Therefore, it can be expected that with the development of new experimental methods, the rate constants may need to be re-evaluated and the mechanistic conclusions adjusted.HNO show a different chemical reactivity than"} +{"text": "The tetrel bond strength for the [I4C\u00b7\u00b7\u00b7X\u2212] series and [I4Tt\u00b7\u00b7\u00b7X\u2212] , was weak-to-moderate, whereas that in the remaining 16 complexes was dative tetrel bond type with very large interaction energies and short Tt\u00b7\u00b7\u00b7X close contact distances. The basis set superposition error corrected interaction energies calculated with the highest-level theory applied, [CCSD(T)/def2-TZVPPD], ranged from \u22123.0 to \u2212112.2 kcal mol\u22121. The significant variation in interaction energies was realized as a result of different levels of tetrel bonding environment between the interacting partners at the equilibrium geometries of the complex systems. Although the \u03c9B97X-D computed intermolecular geometries and interaction energies of a majority of the [I4Tt\u00b7\u00b7\u00b7X\u2212] complexes were close to those predicted by the highest level of theory, the MP2 results were shown to be misleading for some of these systems. To provide insight into the nature of the intermolecular chemical bonding environment in the 25 molecule\u2013anion complexes investigated, we discussed the charge-density-based topological and isosurface features that emanated from the application of the quantum theory of atoms in molecules and independent gradient model approaches, respectively.Twenty-five molecule\u2013anion complex systems [I Ion\u2013molecule interactions are fascinating in chemistry ,2,3,4, b\u2212, Br\u2212, or I\u2212) attracts the electrophiles on bonded Si atoms in the neutral molecules. Neither anion is located precisely at the centroid of the neutral molecule(s). When the anion (Cl\u2212) is entrapped inside the Si20 cage of a fully or partially chlorinated icosasilane molecular entity (20 cage to stabilize the tetrel-centered (Cl\u2212)Si\u00b7\u00b7\u00b7Cl and/or (H\u2212)Si\u00b7\u00b7\u00b7Cl close contacts.r entity e,f, its The fundamental phenomena that drive isolated neutral molecules to self-assemble with anions play a significant role in the processes of anion recognition and anion transport, among others ,17,18. O4, of the tetrel tetrahalide family (TtX4), where Tt stands for the elements in Group 14 of the periodic table and X represents the halide derivative . The anions considered were the halide derivative, X\u2212. It is worth mentioning that the theoretical chemistry of 1:1 complexes formed of lighter members of the TtX4 family with the first three halide anions was recently reported complexes be called ordinary tetrel bonds . Density functional theory ,31,32,33 (QTAIM) ,41,42,43 (QTAIM) ,45-based (QTAIM) ,46 respo(\u03c9B97X-D ) and ab methods, in conjunction with the def2-TZVPPD basis set, were also employed to demonstrate the accuracy of MP2 and \u03c9B97X-D geometries and energies of [I4Tt\u00b7\u00b7\u00b7X\u2212]. Standard non-relativistic calculations were performed without considering the effect of spin-orbit coupling for heavy atoms such as Pb, following a previous study level. An isoelectron density envelope of 0.001 a.u. was used on which to compute the potential, even though the use of higher isoelectron density envelopes was suggested in other studies for chemical systems containing low-polarizable atomic basins are consistent with the topological charge-density-based features emanated using QTAIM. Both Multiwfn . The AIMAll code was used and [\u03c9B97X-D/def2-QZVPPD]. The best agreement between experiment and theory is observed with [\u03c9B97X-D/def2-QZVPPD], and MP2 shows a tendency to underestimate r(Tt\u2013I). A very similar trend was obtained with these methods in conjunction with the def2-QZVPPD basis set.Crystals of TtIe (ICSD) ,78,79,804. From these, it may be seen that the atomic basins are connected to each other in each isolated monomer through bond paths (solid lines between atomic basins in atom color) that pass-through bond critical points (tiny green spheres), thus recovering the expected tetrahedral Td shape of TtI4.b is larger at the C\u2013I bcps in CI4 than at the Pb\u2013I bcps in PbI4. It follows the trend across the series: \u03c1b (C\u2013I) > \u03c1b (Si\u2013I) > \u03c1b (Ge\u2013I) > \u03c1b (Sn\u2013I) > \u03c1b (Pb\u2013I). The trend signifies that the charge concentration is predominant at the C\u2013I bcps in CI4 relative to that at the Pb\u2013I bcps in PbI4.The charge density \u03c1b, b is stabilizing (Hb < 0) at the Tt\u2013I bcps, which is due to the potential energy density Vb that dominates over the gradient kinetic energy density Gb at the bcp. Hb is increasingly more positive at the Tt\u2013I bcps across the series from CI4 through SiI4 to GeI4 to SnI4 to PbI4. This is consistent with the character of Tt\u2013I bonds in TtI4, in which it progressively becomes less covalent than ionic, passing from CI4 through SiI4 to PbI4. That is, the covalency of the Tt\u2013I bond follows this order: C\u2013I > Si\u2013I > Ge\u2013I > Sn\u2013I > Pb\u2013I. Furthermore, the sign of \u22072\u03c1b at Tt\u2013I bcps is also negative for all monomers except for the Tt\u2013I bcps, giving an indication that the Tt\u2013I bonds in CI4, SiI4, and GeI4 are relatively more covalent than those in SnI4 and PbI4. Typically, \u22072\u03c1b < 0 and Hb < 0 represent covalent (shared-type) interactions; \u22072\u03c1b > 0 and Hb > 0 represent ionic (closed-shell) interactions; and \u22072\u03c1b > 0 and Hb < 0 represents mixed interactions complexes. They are shown in The five halide anions have linearly approached the Tt atom from the opposite side of the I\u2013Tt covalent bond in TtI4 with the five halide anions. Because the \u03c3-hole on the carbon atom in CI4 is the weakest compared to that on the Tt atom in TtI4 , the strC\u00b7\u00b7\u00b7At\u2212] , respect4C\u00b7\u00b7\u00b7X\u2212] increases as the halogen derivative becomes more polarizable; it is smallest in [I4C\u00b7\u00b7\u00b7F\u2212], with r(Tt\u00b7\u00b7\u00b7F) = 2.665 \u00c5 with [CCSD/def2-TZVPPD]. By contrast, the I\u2013C\u2013I angle in complexed [I4C\u00b7\u00b7\u00b7F\u2212] either increases or decreases compared to that of its uncomplexed counterpart (\u2220I\u2013C\u2013I = 109.47\u00b0). For instance, the I\u2013C bond linearly attached to the anion forms smaller \u2220I\u2013C\u2013I with the three nearest-neighbor I atoms, whereas the remaining I\u2013C bonds that are not directly involved in the formation of tetrel bond are associated with larger \u2220I\u2013C\u2013I. These two angles types are 107.5 (111.3\u00b0), 108.1 (110.8\u00b0), 108.4 (110.5), 108.8 (110.1\u00b0), and 108.8 (110.1\u00b0) in [I4C\u00b7\u00b7\u00b7F\u2212], [I4C\u00b7\u00b7\u00b7Cl\u2212], [I4C\u00b7\u00b7\u00b7Br\u2212], [I4C\u00b7\u00b7\u00b7I\u2212], and [I4C\u00b7\u00b7\u00b7At\u2212], respectively.The C\u00b7\u00b7\u00b7X intermolecular distance in [I4C\u00b7\u00b7\u00b7F\u2212], 4C\u00b7\u00b7\u00b7F\u2212]. The latter ones mimic the I\u00b7\u00b7\u00b7X links in the other four members of the same family , a feature that has been recommended for identifying hydrogen bonds [4C\u00b7\u00b7\u00b7F\u2212], [I4C\u00b7\u00b7\u00b7Cl\u2212], [I4C\u00b7\u00b7\u00b7Br\u2212], [I4C\u00b7\u00b7\u00b7I\u2212], and [I4C\u00b7\u00b7\u00b7At\u2212] are ca. 2.871, 3.626, 3.892, 4.274, and 4.322 \u00c5, respectively. The former three are less than their respective sum of vdW radii of 3.50 (I + F), 3.86 (I + Cl), and 3.90 \u00c5 (I + Br), whereas the latter two are slightly greater than the sum of their respective sum of vdW radii of 4.08 (I + I) and 4.04 \u00c5 (I + At). . Since rom ref. ,62,97,99re known ,101, whi4C\u00b7\u00b7\u00b7X\u2212], as CCSD. However, the increase in the size of the basis set from def2-TZVPPD to def2-QZVPPD has resulted in a slight increase in the Tt\u00b7\u00b7\u00b7X intermolecular distance with \u03c9B97X-D and MP2. Whatever is the size of the basis set, the intermolecular distances predicted using MP2 are underestimated relative to \u03c9B97X-D and CCSD. Furthermore, [MP2/def2-TZVPPD] has predicted the C\u00b7\u00b7\u00b7I and C\u00b7\u00b7\u00b7At bond distances to be 4.091 and 4.083 \u00c5 for [I4C\u00b7\u00b7\u00b7I\u2212] and [I4C\u00b7\u00b7\u00b7At\u2212], respectively; they were 3.990 and 4.009 \u00c5 with def2-QZVPPD, respectively. This means that MP2 does not correctly predict the trend in C\u00b7\u00b7\u00b7I and C\u00b7\u00b7\u00b7At bonding distances, as predicted by \u03c9B97X-D and CCSD and Hb (Hb < 0) at the bcps of the three equivalent bonds, were equivalent > [I4C\u00b7\u00b7\u00b7Cl\u2212] (\u03b4 = 0.030) > [I4C\u00b7\u00b7\u00b7Br\u2212] (\u03b4 = 0.023) > [I4C\u00b7\u00b7\u00b7I\u2212] (\u03b4 = 0.019) > [I4C\u00b7\u00b7\u00b7At\u2212] (\u03b4 = 0.017), in consistent with the trend found for interaction energy is completely lost.The nature of the intermolecular bonding environment found in c\u2013e. Simil, or Br a\u2013c, but Ge\u00b7\u00b7\u00b7X\u2212] d,e. This4Si\u00b7\u00b7\u00b7Br\u2212], in which, the responsible interacting units were involved in the formation of a dative tetrel bond; it is in a manner similar to that found for [I4Si\u00b7\u00b7\u00b7X\u2212] . On the other hand, MP2 has recognized the attraction between I4Si and X\u2212 in the first four members of the [I4Si\u00b7\u00b7\u00b7X\u2212] series to be unusually strong and that in [I4Si\u00b7\u00b7\u00b7At\u2212] to be moderate. The former result with MP2 is applicable to the [I4Ge\u00b7\u00b7\u00b7X\u2212] series as well. This means that the Tt\u00b7\u00b7\u00b7X close contacts in the above-mentioned molecule\u2013anion systems are not ordinary tetrel bonds; they are dative tetrel bonds.While the bonding features noted above were obtained from [\u03c9B97X-D/def2-QZVPPD], the [CCSD/def2-QZVPPD] method has predicted an exception for [Ib is appreciable at the Ge\u00b7\u00b7\u00b7X bcps in [I4Ge\u00b7\u00b7\u00b7X\u2212] when X points to F, Cl, and Br. For [I4Ge\u00b7\u00b7\u00b7I\u2212], the \u03c1b is small at Ge\u00b7\u00b7\u00b7I bcp (\u03c1b = 0.0048 a.u.), and the interaction between the monomers is also reinforced by I\u00b7\u00b7\u00b7I interactions bcps in [I4Ge\u00b7\u00b7\u00b7X\u2212] are not only typical for coordinate bonds but larger than that can be expected for ordinary non-covalent interactions such as hydrogen bonds, and halogen bonds, among others (\u03c1b < 0.05 a.u.). They may be comparable with the \u03c1b values of the Tt\u2013I coordinate bonds in isolated and complexed TtI4. A similar conclusion might be arrived at for Si\u00b7\u00b7\u00b7X bcps in [I4Si\u00b7\u00b7\u00b7X\u2212] , Our QTAIM analysis, ractions d. The \u03c1b2\u03c1b (\u22072\u03c1b > 0) and Hb (Hb < 0), 4Si\u00b7\u00b7\u00b7X\u2212] provided [CCSD/def2-TZVPPD] results are considered. The large \u03b4 values corresponding to atom\u2013atom pairs responsible for the Si\u00b7\u00b7\u00b7X and Ge\u00b7\u00b7\u00b7X close contacts provide further evidence that there are no \u03c0-type interactions involved; they are purely \u03c3-type coordinate dative bonds. By contrast, the \u03b4 values are very small for atom\u2013atom pairs causing the Si\u00b7\u00b7\u00b7X and Si\u00b7\u00b7\u00b7X close contacts in [I4Ge\u00b7\u00b7\u00b7X\u2212] and [I4Ge\u00b7\u00b7\u00b7X\u2212] , respectively, indicative of closed-shell interactions. The three equivalent I\u00b7\u00b7\u00b7X halogen\u00b7\u00b7\u00b7halogen close contacts in each of [I4Ge\u00b7\u00b7\u00b7X\u2212] and [I4Ge\u00b7\u00b7\u00b7X\u2212] are described by small \u03b4 values, and positive \u22072\u03c1b and Hb. Similarly, the Si\u00b7\u00b7\u00b7X and Ge\u00b7\u00b7\u00b7X tetrel bonds are described by small \u03b4 values, as expected.From the sign and magnitude of \u22074 and PbI4, respectively, are stronger than those of TtI4 . Therefore, their acidic strengths are adequate enough to recognize the five halide anions when in close proximity. This may be rationalized from QTAIM\u2019s molecular graphs of resulting configurations, [I4Sn\u00b7\u00b7\u00b7X\u2212] and [I4Pb\u00b7\u00b7\u00b7X\u2212] , illustrated in 4 and PbI4. This means that the TtI4 molecule is structurally fully deformed in the presence of each of the five halide anions. There is no secondary intermolecular interaction that can play a role in the geometrical stability of the resulting complex anions, as found for other series (see above). In all cases, the tetrel center adopts a trigonal bipyramidal geometry . Clearly, the resulting complex anions each is nothing but a coordination compound, and the Tt\u00b7\u00b7\u00b7X close-contact is formally a Tt\u2013X dative tetrel bond. In such cases, charge transfer from the anion to the \u03c3*(I\u2013Tt) anti-bonding orbital is expected, and the SN2 mechanism is likely to play a role in driving the dative bond formation between the interacting species [The \u03c3-holes on Sn and Pb in SnI species .4Pb\u00b7\u00b7\u00b7X\u2212] series and the smallest for the [I4C\u00b7\u00b7\u00b7X\u2212] series. In particular, the [\u03c9B97X-D/def2-QZVPPD] level QTAIM charge transfer from X\u2212 to PbI4 is 0.263, 0.371, 0.424, 0.515 and 0.588 e for [I4Pb\u00b7\u00b7\u00b7F\u2212], [I4Pb\u00b7\u00b7\u00b7Cl\u2212], [I4Pb\u00b7\u00b7\u00b7Br\u2212], [I4Pb\u00b7\u00b7\u00b7I\u2212], and [I4Pb\u00b7\u00b7\u00b7At\u2212], respectively. The corresponding charge transfer values were 0.229, 0.334, 0.382, 0.460, and 0.519 e for [I4Sn\u00b7\u00b7\u00b7F\u2212], [I4Sn\u00b7\u00b7\u00b7Cl\u2212], [I4Sn\u00b7\u00b7\u00b7Br\u2212], [I4Sn\u00b7\u00b7\u00b7I\u2212], and [I4Sn\u00b7\u00b7\u00b7At\u2212], respectively. These results imply that the nature of charge-transfer in the Sn- and Pb-based anions is virtually similar and that the charge-transfer preference is consistent with the interaction energy preference across a given series, indicating that the charge-transfer phenomenon is likely to be one of the most prominent contributors to the interaction. Our calculation suggests that the extent of charge transfer is the largest for the [Ie calculated for [I4C\u00b7\u00b7\u00b7F\u2212], [I4C\u00b7\u00b7\u00b7Cl\u2212], [I4C\u00b7\u00b7\u00b7Br\u2212], [I4C\u00b7\u00b7\u00b7I\u2212], and [I4C\u00b7\u00b7\u00b7At\u2212], respectively. These results may lead to a conclusion that the formation of stronger complexes accompanies an appreciable amount transfer of charge between the interacting monomers, and is not very surprising is non-negligible; it may be comparable to that of the Tt\u2013I bcps of complexed I4Tt. The Tt\u2013I and Tt\u00b7\u00b7\u00b7X bcps are both characterized by \u22072\u03c1b > 0 and Hb < 0, indicating the presence of a mixed covalent and ionic character. Since Hb becomes increasingly more positive at the Tt\u00b7\u00b7\u00b7X bcp passing from [I4Tt\u00b7\u00b7\u00b7F\u2212] through [I4Tt\u00b7\u00b7\u00b7Cl\u2212] to [I4Tt\u00b7\u00b7\u00b7Cl\u2212] to [I4Tt\u00b7\u00b7\u00b7Br\u2212] to [I4Tt\u00b7\u00b7\u00b7I\u2212] to [I4Tt\u00b7\u00b7\u00b7At\u2212], it is clear that these interactions are less covalent in the same order. The \u22072\u03c1b values are decreasing in the series from [I4Tt\u00b7\u00b7\u00b7F\u2212] through [I4Tt\u00b7\u00b7\u00b7Cl\u2212] to [I4Tt\u00b7\u00b7\u00b7At\u2212], indicating that the Tt\u00b7\u00b7\u00b7F tetrel bond is more ionic than the Tt\u00b7\u00b7\u00b7At tetrel bond. In the case of Tt = Si and Ge, the Tt\u00b7\u00b7\u00b7X bcps show \u22072\u03c1b > 0 and Hb < 0 bond in the respective system. The characteristics of Si\u2013I bonds in I4Si resemble the C\u2013I bonds in I4C.From the molecular graphs of QTAIM in d Hb < 0 . The fou4Tt\u00b7\u00b7\u00b7X\u2212] is considerably larger than what were calculated for ordinary tetrel bonds upon its attractive engagement with the halide anions. For comparison, we note that the \u03b4 values for Tt\u2013I and Tt\u00b7\u00b7\u00b7X coordinate and dative tetrel bonds in [I4Tt\u00b7\u00b7\u00b7X\u2212] are smaller than, and comparable to, those reported for metal\u2013C(O) coordinate bonds ; however, those of Tt\u00b7\u00b7\u00b7X ordinary tetrel bonds are comparable with what were reported for metal\u00b7\u00b7\u00b7metal interactions in [M2(CO)10] and [M3(\u03bc-H)3(CO)12] complexes (Tt = C). The I\u00b7\u00b7\u00b7X closed contacts in several of these systems appeared at larger IGM-inter\u03b4g isovalues (Top). On the other hand, the Tt\u00b7\u00b7\u00b7X close contacts showed up at lower IGM-inter\u03b4g isovalues (Bottom). This is not very surprising since smaller isolvalues are typically necessary for the physical appearances of isosurfaces corresponding to weakly bonded interactions. By contrast, the relatively stronger interactions can be traceable with larger isovalues since charge density around the critical bonding region is generally appreciable. The bluish isosurface originated with large IGM-inter\u03b4g isovalues for [I4C\u00b7\u00b7\u00b7F\u2212] indicates that the attraction between the interacting units is very prominent . The view is also transferable to the [I4Ge\u00b7\u00b7\u00b7X\u2212] systems (not shown).The [I4Sn\u00b7\u00b7\u00b7X\u2212] and [I4Pb\u00b7\u00b7\u00b7X\u2212] , the IGM-inter\u03b4g isosurfaces were visualizable with an isovalue close to 0.055 a.u. 4Pb\u00b7\u00b7\u00b7X\u2212] series. Passing from the left to the right of 4Tt\u00b7\u00b7\u00b7F\u2212] through [I4Tt\u00b7\u00b7\u00b7Cl\u2212] to [I4Tt\u00b7\u00b7\u00b7Br\u2212] to [I4Tt\u00b7\u00b7\u00b7I\u2212] to [I4Tt\u00b7\u00b7\u00b7At\u2212] .In the case of [I4C\u00b7\u00b7\u00b7X\u2212] series and [I4Tt\u00b7\u00b7\u00b7X\u2212] , the int (BSSE)E values for all other molecule\u2013anion systems are much larger than the so-called covalent limit for hydrogen bonds [4Tt\u00b7\u00b7\u00b7X\u2212], it is clear that the complex stability is largely determined by the polarizability of the Tt atom in I4Tt and the halogen derivative. These energies calculated in the range from \u22123.0 to \u2212112.2 kcal mol\u22121 with [CCSD(T)/def2-TZVPPD], \u22121 < int (BSSE)E < \u22125.0 kcal mol\u22121), moderate E < \u221210.0 kcal mol\u22121), strong E < \u221225.0 kcal mol\u22121), very strong E \u2264 \u221240.0 kcal mol\u22121), and ultra-strong (int (BSSE)E >> \u221240.0 kcal mol\u22121 . At the highest level of theory applied, [CCSD(T)/def2-TZVPPD], the weakest and strongest of the [I4Tt\u00b7\u00b7\u00b7X\u2212] systems are found to be [I4C\u00b7\u00b7\u00b7At\u2212] (int (BSSE)E = \u22123.20 kcal mol\u22121) and [I4Si\u00b7\u00b7\u00b7F\u2212] (int (BSSE)E = \u2212112.15 kcal mol\u22121), respectively.Except for the with def2-TZVPPD and def2-QZVPPD, respectively; these are indicative of the fact that the strength of the tetrel bond between Si of I4Si and Br\u2212 is moderate. As mentioned already above, this is not the case with MP2 since the intE (BSSE) for the same system, for instance, with def2-QZVPPD, is predicted to be \u221260.0 kcal mol\u22121; the large intE (BSSE) implies that the attraction between I4Si and Br\u2212 causes the formation of Si\u2013Br dative tetrel bond. This result is consistent with [CCSD(T)/def2-TZVPPD], which has predicted an intE (BSSE) of \u221253.16 kcal mol\u22121 for the same system. Second, the [\u03c9B97X-D/def2-TZVPPD] level intE (BSSE) values for the remaining four systems of the [I4Si\u00b7\u00b7\u00b7X]\u2212 series are in qualitative and quantitative agreement with [CCSD(T)/def2-TZVPPD]. MP2, however, unusually overestimated the interaction energies for [I4Si\u00b7\u00b7\u00b7I\u2212] and [I4Si\u00b7\u00b7\u00b7At\u2212]. The discrepancy between the DFT (or CCSD(T)) and MP2 energies is likely due to the latter method\u2019s misleading prediction of the Si\u00b7\u00b7\u00b7I and Si\u00b7\u00b7\u00b7At close contacts, thus pushing the interacting atoms in [I4Si\u00b7\u00b7\u00b7I\u2212] to be bonded with each other via a dative tetrel bond. These peculiar results indicate that applying the MP2 approach to predict the correct nature of the tetrel bond in molecule\u2013anion complex systems formed by heavier tetrel derivatives in molecular entities should be exercised with caution.From int(BSSE),E between the five members of each series [I4Tt\u00b7\u00b7\u00b7X]\u2212 follows the trend: [I4Tt\u00b7\u00b7\u00b7F\u2212] > [I4Tt\u00b7\u00b7\u00b7Cl\u2212] > [I4Tt\u00b7\u00b7\u00b7Br\u2212] > [I4Tt\u00b7\u00b7\u00b7I\u2212] > [I4Tt\u00b7\u00b7\u00b7At\u2212] . Note that changing the basis set from def2-TZVPPD to def2-QZVPPD somehow restored the [CCSD(T)/def2-TZVPPD] level energy preference at the \u03c9B97X-D level (but not at the MP2 level) when Tt = C, but not when Tt = Pb. One reason for the anomalous change in the preference of energy ordering between [I4Tt\u00b7\u00b7\u00b7I\u2212] and [I4Tt\u00b7\u00b7\u00b7At\u2212] is that the post-HF MP2 method greatly overestimates the BSSE, as well as the electron-electron correlation energy, relative to the DFT and CCSD(T). On the other hand, the CCSD(T) method has properly accounted for electron-electron correlation energy, which ensured the correct preference of stabilization energies among the five members of any given series, [I4Tt\u00b7\u00b7\u00b7X\u2212].The preference of BSSE-corrected interaction energy, t\u00b7\u00b7\u00b7At\u2212] . This isint(BSSE)E on the distance of separation r(Tt\u00b7\u00b7\u00b7X) for 25 molecule\u2013anion complexes, [I4Tt\u00b7\u00b7\u00b7X\u2212], obtained using \u03c9B97X-D, MP2 and CCSD(T). Regardless of the different calculation methods utilized, the dependence was found to be quadratic. The square of the regression coefficient R2 was moderately higher (R2 = 0.9325) for \u03c9B97X-D compared to CCSD(T) and lower (R2 = 0.8923) for MP2.\u22121, from 2.60 to 13.58 kcal mol\u22121, and from 1.42 to 13.44 kcal mol\u22121, respectively. However, when using the def2-QZVPPD basis set, the BSSE in energy has decreased sharply, giving rise to values in the range from 0.02 to 0.13 kcal mol\u22121 with \u03c9B97X-D and from 1.01 to 5.01 kcal mol\u22121 with MP2. CCSD(T) with def2-QZVPPD was computationally very expensive; no conclusions could be drawn about the range of BSSE in energy with this method. int(BSSE)E and intE, obtained using [\u03c9B97X-D/def2-QZVPPD], [MP2/def2-QZVPPD] and [CCSD(T)/def2-TZVPPD], respectively, showing a perfect linear dependence at the former level than that at the latter two. We note further that the BSSE in energy is minimal with DFT but larger with MP2 and CCSD(T). It is very large with the def2-TZVPPD basis set than with the def2-QZVPPD basis set. For example, for \u03c9B97X-D, MP2, and CCSD(T) with def2-TZVPPD, the BSSE in energy ranged from 0.05 to 9.68 kcal mol4Tt\u00b7\u00b7\u00b7X\u2212] was theoretically investigated to clarify the nature of the selectivity of the I4Tt host for five guest anions. The MP2 geometries and interaction energies for the 25 molecule\u2013anion systems were underestimated and overestimated, respectively, relative to DFT and CCSD methods, and in some cases, the MP2 results were unreliable. The chemical bonding features obtained using DFT were consistent with the computationally expensive CCSD and CCSD(T) results, with an exception for [I4Si\u00b7\u00b7\u00b7Br\u2212]. For the latter, the tetrel bonding characteristics predicted by CCSD could not be reproduced by DFT-\u03c9B97X-D. Similarly, the significant overestimation of the interaction energy of [I4Si\u00b7\u00b7\u00b7I\u2212] with MP2 was in sharp disagreement with that computed using \u03c9B97X-D and CCSD(T).In this study, the series systems, except for [I4C\u00b7\u00b7\u00b7X\u2212] and [I4Si\u00b7\u00b7\u00b7X\u2212] . This was the case with [\u03c9B97X-D/def2-TZVPPD] and [\u03c9B97X-D/def2-QZVPPD], but [I4Ge\u00b7\u00b7\u00b7At\u2212] was added to the exclusion list. The result was different from [MP2/def2-QZVPPD] in that it predicted exceptions only for [I4C\u00b7\u00b7\u00b7X\u2212] , and is not surprising given it is an MP2\u2019s tendency to underestimate intermolecular distances. Although these latter two computational methods exclude intermolecular interactions in systems that do not follow the stringent \u201cless than the sum of vdW radii rule,\u201d the exclusion was also consistent with the nature of bond path topology revealed using QTAIM. For example, for [I4C\u00b7\u00b7\u00b7X\u2212] , [I4Tt\u00b7\u00b7\u00b7X\u2212] and [I4Tt\u00b7\u00b7\u00b7X\u2212] , no QTAIM-based bond path topology exists between Tt and X at the [\u03c9B97X-D/def2-QZVPPD] level. This means that QTAIM does not recognize the existence of Tt\u00b7\u00b7\u00b7X tetrel bonding in the host\u2013guest systems when the tetrel bond distance between Tt and X exceeds the sum of the vdW radii of Tt and X, even though this type of limitation of QTAIM has been attributed to the arbitrary nature of the space partitioning scheme. Even so, it should be borne in mind that the Tt and X atoms of the interacting monomers in all complex systems were indeed tetrel bonded to each other, evidenced by the IGM-inter\u03b4g-based isosurfaces.The Tt\u00b7\u00b7\u00b7X separation distance calculated by [CCSD/def2-TZVPPD] was smaller than the vdW radii of the Tt and X atoms for all [I"} +{"text": "The mol\u00adecular packing is consolidated by van der Waals inter\u00adactions between these ribbons.The whole mol\u00adecule of the title compound is generated by twofold rotational symmetry. In the crystal, mol\u00adecules are linked by inter\u00admolecular C\u2014H\u22efO inter\u00adactions with 22H19NO2, is generated by twofold rotational symmetry. The N atom exhibits a trigonal-planar geometry and is located on the twofold rotation axis. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO contacts with R22(12) ring motifs, and C\u2014H\u22ef\u03c0 inter\u00adactions, resulting in ribbons along the c-axis direction. van der Waals inter\u00adactions between these ribbons consolidate the mol\u00adecular packing. Hirshfeld surface analysis indicates that the greatest contributions to the crystal packing are from H\u22efH (45.5%), C\u22efH/H\u22efC (38.2%) and O\u22efH/H\u22efO (16.0%) inter\u00adactions.The whole mol\u00adecule of the title compound, C The phenyl ring (C1\u2013C4/C2A/C3A) attached to the N1 atom and the phenyl rings (C7\u2013C12 and C7A\u2013C12A) of the two symmetry-related 1-phenyl\u00adethan-1-one groups are oriented at 89.65\u2005(6)\u00b0 to each other.The asymmetric unit of the title compound contains half a mol\u00adecule, the complete mol\u00adecule being generated by the twofold rotational axis. Atoms N1, C1 and C4 are located on the twofold rotation axis Fig.\u00a01. The N1 3.A\u22efO1 to 1.2546 (blue) a.u reveal that H\u22efH (45.5%), C\u22efH/H\u22efC (38.2%) and O\u22efH/H\u22efO (16.0%) inter\u00adactions make the greatest contributions to the surface contacts. N\u22efH/H\u22efN (0.3%) contacts also contribute to the overall crystal packing of the title compound. The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, C\u22efH/H\u22efC and O\u22efH/H\u22efO inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing sulfon\u00adyl]-N-phenyl\u00adbenzene\u00adsulfonamide bis\u00ad(pyridine-2-carboxamide) bis\u00ad(3-phenyl\u00adacrylate) [XEBWUY (III); Sabari et al., 2012I) possesses twofold rotational symmetry. The N atom has a trigonal-planar geometry and is located on the twofold rotation axis. Weak C\u2014H\u22efO hydrogen bonds connect the mol\u00adecules, forming a three-dimensional network. The asymmetric unit of (II) contains two independent mol\u00adecules with similar conformations. In the crystal, N\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular structure. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions are also observed. In (III), the C=C double bonds adopt an E configuration. In the crystal, pairs of C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into inversion dimers.A search of the Cambridge Structural Database = 1.2Ueq(C). Owing to poor agreement between observed and calculated intensities, eighteen outliers were omitted during the final refinement cycle.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022005382/tx2050sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022005382/tx2050Isup2.hklStructure factors: contains datablock(s) I. DOI: 2173928CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the mol\u00adecules are cross-linked via inter\u00admolecular C\u2014H\u22efN hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions into a three-dimensional network.In the title compound, the mol\u00adecule adopts an 33H33N3, is a carbazolophane, which is a cyclo\u00adphane composed of two carbazole fragments. It has a planar chirality but crystallizes as a racemate in the space group Panti-configuration, in which two carbazole fragments are partially overlapped. Both carbazole ring systems are slightly bent, with the C atoms at 3-positions showing the largest deviations from the mean planes. The dihedral angle between two carbazole fragments is 5.19\u2005(3)\u00b0, allowing an intra\u00admolecular slipped \u03c0\u2013\u03c0 inter\u00adaction [Cg\u22efCg = 3.2514\u2005(8)\u2005\u00c5]. In the crystal, the mol\u00adecules are linked via inter\u00admolecular C\u2014H\u22efN hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions into a network sheet parallel to the ab plane. The mol\u00adecules of different sheets form other C\u2014H\u22ef\u03c0 inter\u00adactions, thus forming a three-dimensional network.The title compound, C Previously, our group reported the EPR spectrum of the title compound, but no other chemical properties were examined because of the very low yield \u2005\u00c5 for the N1/C4\u2013C15 ring system and 0.062\u2005(1)\u00c5 for N2/C16\u2013C27 ring system. In both carbazole fragments, the C atoms at the 3-positions bridged through the di\u00admethyl\u00adene\u00adamino group show the largest deviations from the mean planes [0.1177\u2005(14)\u2005\u00c5 for C7 and \u22120.1082\u2005(14)\u2005\u00c5 for C19]. The dihedral angle formed by two carbazole fragments is 5.19\u2005(3)\u00b0, providing an intra\u00admolecular slipped parallel \u03c0\u2013\u03c0 inter\u00adaction . In comparison, in the related PO compounds, the dihedral angles between two carbazole fragments and the centroid\u2013centroid distances are 5.96\u2005(6)\u00b0 and 3.294\u2005(4)\u2005\u00c5 for N-cyanamide-bridged [3.3]carbazolophane carbazolophane \u00b0, smaller than those in the related compounds , that is, the hybridization of N3 atom is closer to sp3 than to sp2, reflecting the difference in the resonance effect of the substituent at the N3 atom.The title compound has a planar chirality but crystallizes as a racemate in the centrosymmetric space group 3.C(9) chain motif running parallel to the b axis. The mol\u00adecules are further joined into columns along the a-axis direction by pairs of C\u2014H\u22ef\u03c0 inter\u00adactions , forming a centrosymmetric dimer forms another centrosymmetric dimer -10,11,22,23-tetra\u00adhydro-9H,21H-5,8:15,12-bis\u00ad(metheno)tri\u00adaza\u00adcyclo\u00adhexa\u00addecino-di\u00adindole carbazolophane, YUKYEL; Tani et al., 2020anti-3-cyano-3-aza-1,3-dicarbazola\u00adcycloocta\u00adphane carbazolophane, BACKOG; Tani et al., 2001N-cyanamide-bridged FO [3.3]carbazolophane, syn-3-cyano-3-aza-1,3-dicarbazola\u00adcyclo\u00adocta\u00adphane benzene clathrate \u2005\u00c5 for the benzene rings bridged by cyanamide, 3.447\u2005(2)\u2005\u00c5 for the central pyrrole rings and 3.792\u2005(3)\u2005\u00c5 for the outer benzene rings. Three of the remaining four structures are PO [m.n]carbazolophanes; anti-ethenylene and 1,3-xylylene-bridged [2.5]carbazolophane carbazolophane carbazolophane \u00b0 and 3.8062\u2005(15)\u2005\u00c5 for ethenylene and 1,3-xylylene-bridged [2.5]carbazolophane; 15.04\u2005(9)\u00b0 and 3.732\u2005(3)\u2005\u00c5 for cyanamide-bridged [3.4]carbazolophane; 24.87\u2005(11)\u00b0 and 3.901\u2005(3)\u2005\u00c5 for oxa-bridged [3.5]carbazolophane. The last of the seven compounds is a FO carbazolophane, syn-cyclo\u00adbutane-bridged [2.4]carbazolophane in di\u00adchloro\u00admethane (150\u2005mL) and sodium hydroxide in water (10\u2005mL). Then, the flask was filled with argon and was stirred at room temperature for 3\u2005d. The reaction mixtures were washed with water, then dried over anhydrous sodium sulfate. Solvent was removed under reduced pressure, and the residue was purified by silica gel chromatography . Elution from hexa\u00adne:ethyl acetate (19:1) gave a white solid . Elution from hexa\u00adne:ethyl acetate (10:1) gave mixtures including a syn-configuration (FO isomer), but they were difficult to separate. A part of the title compound was recrystallized from di\u00adchloro\u00admethane:ethanol (1:3) to give a colorless crystal suitable for X-ray diffraction. Melting point: 482\u2013484\u2005K. 1H NMR \u03b4 = 1.08 , 1.57\u20131.60 ,1.78 , 2.81\u20132.97 , 3.74\u20133.90 , 4.10\u20134.17 , 5.34 , 6.38 , 7.25-7.30 , 7.46\u20137.53 , 7.67 , 8.11 .A solution of 9,9\u2032-bis\u00ad[3-(bromo\u00admeth\u00adyl)-96.Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022003383/yk2167sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989022003383/yk2167Isup2.hklStructure factors: contains datablock(s) I. DOI: 2161895CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "IIIS2N2O2 chromophore contains two O,N,S-donating dianionic ligands in perpendicular planes, with the O and S atoms in cis and the N atoms in trans positions. The FeIII ion is in the high-spin state at 100\u2005K. The variable-tem\u00adper\u00adature magnetic susceptibility measurements (5\u2013320\u2005K) are consistent with the presence of a high-spin S\u00a0= 5/2 FeIII ion.The Fe 3)2NH2][Fe(C10H11O2N3S)2], are reported. The asymmetric unit consists of an octa\u00adhedral [FeIII(L)2]\u2212 fragment, where L2\u2212 is 3-eth\u00adoxy\u00adsalicyl\u00adaldehyde thio\u00adsemi\u00adcar\u00adba\u00adzon\u00adate(2\u2212), and a di\u00admethyl\u00adammonium cation. Each L2\u2212 ligand binds with the thiol\u00adate S, the imine N and the phenolate O atoms as donors, resulting in an FeIIIS2N2O2 chromophore. The ligands are orientated in two perpendicular planes, with the O and S atoms in cis positions, and mutually trans N atoms. The FeIII ion is in the high-spin state at 100\u2005K. The variable-tem\u00adper\u00adature magnetic susceptibility measurements (5\u2013320\u2005K) are consistent with the presence of a high-spin FeIII ion with D\u00a0= 0.83\u2005(1)\u2005cm\u22121 and g\u00a0= 2.The synthesis and crystal structure (100\u2005K) of the title com\u00adpound, [(CH IR spectroscopic measurements of (I)\u22121 were carried out at room tem\u00adper\u00adature using an ATR PerkinElmer FT\u2013IR Frontier spectrometer.A room-tem\u00adper\u00adature IR spectrum of 3-eth\u00adoxy\u00adsalicyl\u00adalde\u00adhyde thio\u00adsemicarbazone within the range 4000\u2013400\u2005cm1H and 13C NMR spectra were recorded in DMSO-d6 (dimethyl sulfoxide) using a Bruker cryomagnet BZH 300/52 spectrometer (300\u2005MHz), with the recorded chemical shifts in \u03b4 (in parts per million) relative to an inter\u00adnal standard of tetra\u00admethyl\u00adsilane (TMS).M, versus tem\u00adper\u00adature, T, were conducted between 5 and 320\u2005K, heating and cooling at a rate of 2\u2005K\u2005min\u22121 in an applied field, \u03bc0H, of 0.1\u2005T using a Quantum Design MPMS-5S super\u00adconducting quantum inter\u00adference device (SQUID) magnetometer. The SQUID magnetometer was calibrated using a standard palladium sample. The background due to the sample holder and the diamagnetic signal of the sample, estimated using Pascal\u2019s constants magnetic susceptibility, \u03c72-3-OEt-thsa) was carried out according to the general procedure described by Yemeli Tido : \u03b4 (ppm) 11.39 , 9.02 , 8.40 , 7.90\u20138.13 , 6.72\u20137.50 , 4.05 , 1.35 . 13C NMR : \u03b4 (ppm) 182.8 (C=S), 147.4, 146.7 (C\u2014O), 140.1 (C=N), 119.5, 118.7, 114.5 (C aromatic), 64.6 (C\u2014N), 74.0 (O\u2014CH2), 15.1 (O\u2014C\u2014CH3). IR : 3400 (\u03bdOH), 3169 (\u03bdNH), 3249 (\u03bdNH2), 2935 (\u03bdCH3), 2896 (\u03bdCH2), 1618 (\u03bdC=N), 1535\u20131600 (\u03bdC=C), 1270 (\u03bdC\u2014N), 1167 (\u03bdC=S).The synthesis of 3-eth\u00adoxy\u00adsalicyl\u00adaldehyde thio\u00adsemi\u00adcar\u00adba\u00adzone (HTido 2010 (yield: 3)2NH2][Fe(3-OEt-thsa)2], (I)3)3\u00b79H2O was dissolved in water (10\u2005ml). The ligand H2-3-OEt-thsa was dissolved in methanol (60\u2005ml) with the addition of di\u00admethyl\u00adamine, 40\u2005wt% in water . To this mixture, the FeIII salt solution was added dropwise with constant stirring. The resulting dark-green solution was stirred and heated to 80\u2005\u00b0C for approximately 10\u2005min. The solution was then allowed to stand at room tem\u00adper\u00adature until crystals had formed. The dark-green microcrystals were isolated by filtration and dried . IR : 3436, 3414 (\u03bdNH), 3265, 3098 (\u03bdNH2), 3012 (\u03bdCH3), 2971 (\u03bdCH2), 1614, 1586 (\u03bdC=N), 1570\u20131541 (\u03bdC=C ring), 1238 (\u03bdC\u2014O), 1215 (\u03bdN\u2014N), 1078 (\u03bdC\u2014N), 736 (\u03bdC\u2014S).The synthesis of ferrate(III), (I)P21/n, with Z\u00a0= 4. The asymmetric unit consists of one formula unit, [(CH3)2NH2][Fe(3-OEt-thsa)2], with no atom on a special position. The FeIII cation is coordinated by the thiol\u00adate S, phenolate O and imine N atoms of each of the two dianionic O,N,S-tridentate L2\u2212 ligands. The donor atoms of the ligands are situated in two perpendicular planes, with the O and S atoms in cis positions, and mutually trans N atoms. Selected geometric parameters are listed in Table\u00a02The structure of di\u00admethyl\u00adammonium bis\u00ad com\u00adpound at 103 (and 298\u2005K) confirm that the FeIII cation can be related to the bond angles of the FeO2N2S2 coordination core. An analysis of the bond angles involving the opposite ligand donor atoms at 100\u2005K is very enlightening, as it shows that the octa\u00adhedral geometry of the present high-spin FeIII com\u00adpound, with O1\u2014Fe1\u2014S1\u00a0= 158.48\u2005(5)\u00b0, O101\u2014Fe1\u2014S101\u00a0= 158.89\u2005(5)\u00b0 and N1\u2014Fe1\u2014N101\u00a0= 167.63\u2005(7)\u00b0, is considerably less regular than that of the low-spin com\u00adpound Cs[Fe(3-OEt-thsa-Me)2]\u00b7CH3OH, with the bond angles S11\u2014Fe\u2014O11\u00a0= 177.83\u2005(14)\u00b0, S21\u2014Fe\u2014O21\u00a0= 178.01\u2005(13)\u00b0 and N11\u2014Fe\u2014N21\u00a0= 178.9\u2005(2)\u00b0 \u00b0 and O101\u2014Fe\u2014N101\u00a0= 84.03\u2005(7)\u00b0] than the five-mem\u00adbered chelate ring [S1\u2014Fe\u2014N1\u00a0= 78.45\u2005(5)\u00b0 and S101\u2014Fe\u2014N101\u00a0= 78.93\u2005(5)\u00b0]. The r.m.s. deviations from their least-squares plane of atoms of the six-mem\u00adbered chelate ring of both coordinated ligands are 0.197 and 0.177\u2005\u00c5 for Fe1/N11/C17/C11/C12/O11 and Fe1/N101/C107/C101/C102/O101, respectively, and the corresponding values for the five-mem\u00adbered chelate rings are 0.129 and 0.102\u2005\u00c5 for Fe1/N11/C12/C18/S11 and Fe1/N101/C102/C108/S101, respectively. It appears that the metal chelate rings deviate slightly from the ideal planar structure. Furthermore, the O\u2014Fe\u2014N and S\u2014Fe\u2014N bite angles of the six- and five-mem\u00adbered chelates are deficient by ca 38 and 30\u00b0, respectively, com\u00adpared to the angle at the vertex of a regular hexa\u00adgon (120\u00b0) or penta\u00adgon (108\u00b0), respectively. In com\u00adparison to other (cation+)[Fe(L2\u2212)2]\u00b7x(solvent) com\u00adpounds of related ligands com\u00adpounds contain FeIII in the low-spin state, whereas the present com\u00adpound contains FeIII in the high-spin state. Consequently, (I) displays longer FeIII\u2013donor atom bond lengths, which are associated with more restricted bite angles. Moreover, the remaining bond angles involving each six-mem\u00adbered chelate ring or the thiol\u00adate S (S1 and S101) atoms. The charge of the two g Table\u00a02 are, as III com\u00adplex is further enhanced by the high degree of electron delocalization throughout the chelated ligands, which is evident from the geometric parameters. The C\u2014S, C\u2014N and N\u2014N bond lengths of (I)i.e. single bond) and 2 (i.e. double bond). The C8\u2014S1 bond length of 1.746\u2005(3)\u2005\u00c5 and the C108\u2014S101 bond length of 1.752\u2005(2)\u2005\u00c5 suggest partial electron delocalization of these C\u2014S bonds. This feature has also been found in the structure of the related high-spin FeIII com\u00adpound Cs[Fe(thsa)2] at 103\u2005K at 100\u2005K of 1.301\u2005(3) and 1.301\u2005(3)\u2005\u00c5, respectively, which correspond to the C\u2014N bond lengths of 1.314\u2005(10) and 1.303\u2005(11)\u2005\u00c5, respectively, for Cs[Fe(thsa)2] at 103\u2005K OLEX2 versus tem\u00adper\u00adature measurements for (I)III ion. The data collected on heating and cooling coincide over the tem\u00adper\u00adature range studied. The tem\u00adper\u00adature dependence of \u03c7MT collected on cooling between 320 and 5\u2005K is displayed in Fig.\u00a03MT is tem\u00adper\u00adature independent with a value of 4.41\u2005(1)\u2005cm3\u2005K\u2005mol\u22121 [5.94\u2005(1)\u2005\u00b5B/Fe]. This is just above the expected value of 4.38\u2005cm3\u2005K\u2005mol\u22121 (5.92\u2005\u00b5B/Fe) for FeIII in its high-spin (S\u00a0= 5/2) state with an electronic g factor of 2. \u03c7M\u22121(T) is linear in T and a fit to a Curie\u2013Weiss law between 100 and 320\u2005K shown in Fig.\u00a04B/Fe.Magnetic susceptibility MT drops rapidly below 100\u2005K. This may be due to weak (anti\u00adferro)magnetic inter\u00adactions between neighbouring spins or may reflect a splitting of the S\u00a0= 5/2 state term HCEF\u00a0= D[zS2 \u2212 S(S\u00a0+\u00a01)/3] + E(xS2 \u2013 yS2), with D and E being the axial and rhombic zero-field splitting, respectively. The 6S high-spin state is split into three Kramers doublets. For E\u00a0= 0, the doublets are separated by 2D and 6D from the lowest energy doublet. The Zeeman energy zH\u00a0= g\u03bcBzHS and the molar susceptibility with a field along z isX\u00a0= D/kBT, NA is Avogadro\u2019s number and kB is the Boltzmann constant \u2005cm\u22121 with g\u00a0= 2. D is in the range expected for high-spin FeIII spectroscopy.\u03c7 al. 2002, but theIII com\u00adpounds that have so far been reported to contain the 3-eth\u00adoxy\u00adsalicyl\u00adaldehyde 4-R\u2032-thio\u00adsemi\u00adcar\u00adba\u00adzon\u00adate(2\u2212) dianion. In Cs[Fe(3-OEt-thsa-Me)2]\u00b7CH3OH 2NH2][Fe(3-OEt-thsa)2] com\u00adpound, (I)R\u2032 substituent on the terminal N atom of the thio\u00adsemicarbazide moiety, as (I) contains a H atom, whereas Cs[Fe(3-OEt-thsa-Me)2]\u00b7CH3OH 2NH2+versus Cs+; and (iii) the presence of a methanol solvent mol\u00adecule in the crystal lattice of Cs\u00ad[Fe(3-OEt-thsa-Me)2]\u00b7CH3OH , whereas Cs\u00ad[Fe(3-OEt-thsa-Me)2]\u00b7CH3OH forms inter\u00admole\u00adcular hydro\u00adgen-bonded ring systems which link neighbouring FeIII entities. These factors determine the arrangement of the FeIII entities within the unit cell, which is further characterized by the space group P21/n, with Z\u00a0= 4 and V\u00a0= 2576.35\u2005(17)\u2005\u00c53 for (I), with a volume of 644.09\u2005\u00c53 per high-spin FeIII formula unit, and the space group PZ\u00a0= 2 and V\u00a0= 1369.5\u2005(8)\u2005\u00c53 for Cs[Fe(3-OEt-thsa-Me)2]\u00b7CH3OH, with a volume of 684.75\u2005\u00c53 per low-spin FeIII formula unit [Fe(L2\u2212)2]\u00b7x(solvent) materials and allows for a variation of the spin state of FeIII, with some members dis\u00adplaying tem\u00adper\u00adature-dependent spin-crossover behaviour I, global. DOI: 10.1107/S2053229622011597/jx3075Isup2.hklStructure factors: contains datablock(s) I. DOI: 2223775CCDC reference:"} +{"text": "C(8) chains along the [100] direction. C\u2014Br\u22ef\u03c0 inter\u00adactions connect these chains into parallel layers to (002). van der Waals inter\u00adactions between the layers aid in the cohesion of the crystal packing.C\u2014H\u22efBr inter\u00adactions connect mol\u00adecules in the crystal, resulting in zigzag 14H8Br3N3O2, consists of three almost planar groups: the central di\u00adbromo\u00adethenyldiazene fragment and two attached aromatic rings. The mean planes of these rings form dihedral angles with the plane of the central fragment of 26.35\u2005(15) and 72.57\u2005(14)\u00b0 for bromine- and nitro-substituted rings, respectively. In the crystal, C\u2014H\u22efBr inter\u00adactions connect mol\u00adecules, generating zigzag C(8) chains along the [100] direction. These chains are linked by C\u2014Br\u22ef\u03c0 inter\u00adactions into layers parallel to (001). van der Waals inter\u00adactions between the layers aid in the cohesion of the crystal packing. The most substantial contributions to crystal packing, according to a Hirshfeld surface analysis, are from Br\u22efH/H\u22efBr (20.9%), C\u22efH/H\u22efC (15.2%), O\u22efH/H\u22efO (12.6%) and H\u22efH (11.7%) contacts.The mol\u00adecule of the title compound, C The crystal packing is consolidated by C\u2014F\u22ef\u03c0 contacts and \u03c0\u2013\u03c0 stacking inter\u00adactions, and short Br\u22efO [2.9828\u2005(13)\u2005\u00c5] distances are also observed. In the crystal of (III), the mol\u00adecules are linked into inversion dimers via short halogen\u2013halogen contacts compared to the van der Waals radius sum of 3.50\u2005\u00c5 for two chlorine atoms. No other directional contacts could be identified, and the shortest aromatic ring centroid separation is greater than 5.25\u2005\u00c5. In the crystals of (IV) and (V), the mol\u00adecules are linked through weak X\u22efCl contacts [X = Cl for (IV) and Br for (V)], C\u2014H\u22efCl and C\u2014Cl\u22ef\u03c0 inter\u00adactions into sheets lying parallel to (001). In the crystal of (VI), the mol\u00adecules are stacked in columns parallel to [100] via weak C\u2014H\u22efCl hydrogen bonds and face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions. The crystal packing is further consolidated by short Cl\u22efCl contacts. In (VII), mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds into zigzag chains running parallel to [001]. The crystal packing also features C\u2014Cl\u22ef\u03c0, C\u2014F\u22ef\u03c0 and N\u2014O\u22ef\u03c0 inter\u00adactions. In (VIII), C\u2014H\u22efN and short Cl\u22efCl contacts are observed, and in (IX), C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and short Cl\u22efO contacts occur.In -1-(4-bromo\u00adphen\u00adyl)-2-(4-nitro\u00adbenzyl\u00adidene)hydrazine (1\u2005mmol), tetra\u00admethyl\u00adethylene\u00addi\u00adamine , CuCl and CBr4 (4.5\u2005mmol). After 1-3\u2005h , the reaction mixture was poured into 0.01 M solution of HCl , and extracted with di\u00adchloro\u00admethane (3 \u00d7 20\u2005mL). The combined organic phase was washed with water (3 \u00d7 50\u2005mL), brine (30\u2005mL), dried over anhydrous Na2SO4 and concentrated in vacuo using a rotary evaporator. The residue was purified by column chromatography on silica gel using appropriate mixtures of hexane and di\u00adchloro\u00admethane (3/1\u20131/1). Crystals suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution. Red solid (58%); m.p. 398\u2005K. Analysis calculated for C14H8Br3N3O2 (M = 489.95): C 34.32, H 1.65, N 8.58; found: C 34.27, H 1.70, N 8.56%. 1H NMR \u03b4 8.16\u20137.41 . 13C NMR \u03b4 150.89, 149.62, 148.26, 136.43, 132.25, 127.77, 125.57, 124.53, 123.57, 93.24. ESI\u2013MS: m/z: 490.96 [M + H]+.This dye was synthesized according to the reported method (Akkurt 6.Uiso(H) = 1.2Ueq(C). One reflection (110), affected by the beam stop, was omitted in the final cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698902200620X/yk2170sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902200620X/yk2170Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902200620X/yk2170Isup3.cmlSupporting information file. DOI: 2178832CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The synthesis and structures of dinuclear palladium complexes with 1,3-benzimidazolidine-2-thione and 1,3-imidazoline-2-thione are reported. H-benzimidazole-2-thiol\u00adato)-\u03ba2N3:S;\u03ba2S:N3-bis\u00ad[cyanido(tri\u00adphenyl\u00adphosphine-\u03baP)palladium(II)], [Pd2(C7H5N2S)2(CN)2(C18H15P)2] or [Pd22(CN)2(PPh3)2] (1), and bis\u00ad(\u03bc-1H-imidazole-2-thiol\u00adato)-\u03ba2N3:S;\u03ba2S:N3-bis\u00ad[cyanido(tri\u00adphenyl\u00adphosphine-\u03baP)palladium(II)] aceto\u00adnitrile 0.58-solvate, [Pd2(C3H3N2S)2(CN)2(C18H15P)2]\u00b70.58C2H3N or [Pd22(CN)2(PPh3)2]\u00b70.58C2H3N (2). The compound [Pd22(CN)2(PPh3)2] is located on a crystallographic twofold axis while [Pd22(CN)2(PPh3)2]. 0.58(C2H3N) contains two partially occupied aceto\u00adnitrile solvent mol\u00adecules with occupancies of 0.25 and 0.33. In both of these compounds, the anionic bzimtH\u2212 and imtH\u2212 ligands coordinate through N,S-donor atoms in a bridging mode, covering four coordination sites of two metal centers and other two sites are occupied by two PPh3 ligand mol\u00adecules. Finally, the remaining two sites of two metal centers are occupied by cyano groups, abstracted by the metals from the solvent during reaction. In the packing of the 1,3-benzimidazolidine- 2-thione and 1,3-imidazoline-2-thione complexes, there are intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions involving the thione moiety as well as an N\u2014H\u22efN hydrogen bond linking the thione and cyano ligands. In addition, in 2, as well as the \u03c0\u2013\u03c0 inter\u00adaction involving the thione moieties, there is an additional \u03c0\u2013\u03c0 inter\u00adaction involving one of the thione moieties and an adjacent phenyl ring from the tri\u00adphenyl\u00adphosphine ligand. There are also C\u2014H\u22efN inter\u00adactions between the imidazoline rings and the aceto\u00adnitrile N atoms.The synthesis and structures of dinuclear palladium complexes with 1,3-benz\u00adimidazolidine-2-thione (bzimtH) and 1,3-imidazoline-2-thione (imtH) are reported, namely, bis\u00ad(\u03bc-1 However, the X-ray crystal structure of the product revealed the formation of the unexpected dinuclear compound [Pd22(CN)2(PPh3)2] (1). Another thio-ligand, imtH2 yielded a similar dinuclear compound, [Pd22-(CN)2(PPh3)2] (2). In both these compounds, the anionic bzimtH\u2212 and imtH\u2212 ligands coordinate through N,S donor atoms in a bridging mode, covering four coordination sites of two metal centers, and other two sites are occupied by two PPh3 ligand mol\u00adecules. Finally, the remaining two sites of two metal centers are occupied by cyano groups, abstracted by the metals from the solvent during reaction.The reaction of PdCl1 crystallizes in the monoclinic space group C2/c, and compound 2 in the monoclinic space group, P21/c. Selected bond distances and bond angles are given in Tables 11 is shown in Fig.\u00a012 is shown in Fig.\u00a021, here only half of the mol\u00adecule is unique as the mol\u00adecule lies on a crystallographic twofold axis. In 1, the Pd metal atom is bonded to one P, one S, one N and one C atoms with the respective bond distances as follows: Pd\u2014P = 2.2861\u2005(6), Pd\u2014S = 2.3547\u2005(6), Pd\u2014N = 2.0545\u2005(17), and Pd\u2014C = 1.959\u2005(2)\u2005\u00c5. The trans bond angles, P\u2014Pd\u2014S and N\u2014Pd\u2014C, of 172.26\u2005(2) and 178.31\u2005(8)\u00b0, as well as the cis bond angles in the range 84.93\u2005(6)\u201394.24\u2005(5)\u00b0, reveal the distorted square-planar geometry of each metal center. One of the major factors in the conformation adopted by the mol\u00adecule is the strong \u03c0\u2013\u03c0 inter\u00adaction between the thione moieties , as seen in Fig.\u00a01Cg\u22efCg = 3.3560\u2005(9)\u2005\u00c5 with a slippage of 1.408\u2005\u00c5].Compound 2 is similar to that of 1. Nevertheless, there are minor differences in the bond distances and angles pertaining to the two metal centers of compound 2 , 2.3541\u2005(5), 2.0345\u2005(17) and 1.957\u2005(2)\u2005\u00c5 , and 2.2984\u2005(5), 2.3542\u2005(5), 2.0345\u2005(17) and 1.943\u2005(2)\u2005\u00c5 . For both metal centers, the trans bond angles [P\u2014Pd\u2014S and N\u2014Pd\u2014C = 173.86\u2005(2)\u2013179.31\u2005(8)\u00b0] and the adjacent bond angles [86.54\u2005(7)\u2013 92.80\u2005(4)\u00b0] are similar to those of compound 1. These bond angles again reveal the distorted square-planar geometry of each metal center of compound 2. The various bond distances described above are normal and none unusual. Compound 1 has carbon\u2013sulfur (C\u2014S) bond distance of 1.728\u2005(2), while in compound 2 it is 1.734\u2005(2)\u2005\u00c5. These distances are in between single (1.81\u2005\u00c5) and double-bond (1.68\u2005\u00c5) C\u2014S distances /1.148\u2005(3)\u2005\u00c5 in compound 2. These distances are less than the expected C=N double bond (1.28\u2005\u00c5) and are close to the C\u2261N triple bond distance \u2005\u00c5], as seen in Fig.\u00a02Cg\u22efCg distances of 3.3065\u2005(8)\u2005\u00c5 and 3.3218\u2005(8), respectively, with a slippage for the latter of 1.154\u2005\u00c5].The coordination pattern of compound 2 Fig.\u00a02. Thus, t2 ligand showed a \u03bd(N\u2014H) band at 3113 (m), and in compound 1, this band appeared at a lower energy, 3055 (m) cm\u22121. The ligand showed a diagnostic \u03bd(C=S) band at 1179\u2005cm\u22121, which shifted to \u03bd(C=S), 1033(s) cm\u22121, owing to the change of neutral bzimtH2 ligand to the bzimtH\u2212 anionic form, coordinating through N,S donor atoms. The PPh3 ligand showed its characteristic \u03bd(P\u2014CPh) band at 1097(s) cm\u22121 in compound 1. A band at 1734\u2005cm\u22121 was assigned to the coordinated cyano group. The IR spectroscopic bands of compound 2 are similarly assigned: \u03bd(N\u2014H), 3050 (m), \u03bd(C=S), 1020 (m), \u03bd(P\u2014CPh), 1105 (s) and \u03bd(C\u2261N), 1740(s) cm\u22121.The IR spectrum of the bzimtH2) has formed an N,S-bonded symmetrically bridged dinuclear compound, and so is the case with 1,3-imidazolidine-2-thione, and these are analogous to literature reports , 1,3-benzimidazoline-2-thione (bzimtH2), 1,3-imidazoline-2-thione (imtH2), and tri\u00adethyl\u00adamine were procured from Aldrich. The solvents were of HPLC grade and were stored over mol\u00adecular sieves. The precursor, PdCl2(PPh3)2, was prepared by a literature procedure chloride, tri\u00adphenyl\u00adphosphine (PPh2(PPh3)2]Preparation of the precursor, [PdClin vacuo, m.p. 551-553\u2005KPalladium(II) chloride was dissolved in hot aceto\u00adnitrile (25\u2005mL) in a 50\u2005mL round-bottom flask, and to it was added tri\u00adphenyl\u00adphosphine . The contents were refluxed for 1\u2005h and the yellow complex formed was filtered and dried 1Preparation of 2(PPh3)2 in 10\u2005mL of CH3CN, was added solid bzimtH2 followed by the addition of Et3N base (0.5\u2005mL). The solution became yellowish orange and was refluxed for 6\u2005h. The orange compound was formed on refluxing. It was separated and dissolved in a solution of methanol (4\u2005mL) and di\u00adchloro\u00admethane (1\u2005mL) in a culture tube. A slow evaporation of the reaction mixture over a period of one month, resulted in the formation of orange crystals of compound 1. Yield: 0.015\u2005g; 65%; m.p. 511\u2013513\u2005K. Analysis found: C, 57.71; H, 3.84; N, 7.50; C52H40N6P2Pd2S2 (1087.8) requires: C, 57.40; H, 3.70; N, 7.72%. IR Data : \u03bd(N\u2014H), 3055 (m); \u03bd(C\u2013H), 2950 (m), 2920 (s), 2852 (m); \u03bd(C\u2261N), 1734 (s), \u03bd(C\u2014C) + \u03b4(N\u2014H) + \u03b4(C\u2014H), 1635 (m), 1440 (s), 1380 (m); \u03bd(P\u2014CPh), 1097 (s); \u03bd(C=S), 1033 (s). Ligand IR Data: \u03bd(N\u2014H), 3113 (m), \u03bd(C\u2014H), 3078 (m); 2981 (s); \u03bd(C\u2261N) 1513 (s), \u03b4(N\u2014H), 1467 (s), 1381 (m); \u03bd(C=S), 1179 (s). The compound is partially soluble in di\u00adchloro\u00admethane, but soluble in methanol and chloro\u00adform.To a solution of PdCl2Preparation of 2(PPh3)2 in 10\u2005mL of CH3CN, was added solid imtH2 followed by the addition of Et3N base (0.5\u2005mL). The solution became yellowish orange and was refluxed for 6\u2005h. The orange compound was formed on refluxing and was separated. It was dissolved in a solution of methanol (4\u2005mL) and di\u00adchloro\u00admethane (1\u2005mL) in a culture tube. Slow evaporation of the reaction mixture over a period of one month formed yellowish-orange crystals of compound 2. Yield: 0.020\u2005g; 69%; m.p. 485\u2013488\u2005K. Analysis found: C, 53.21; H, 3.92; N, 8.48; C44H36N6P2Pd2S2\u00b70.58(CH3CN) (1011.5) requires: C, 53.58; H, 3.73; N, 8.36%. IR bands : \u03bd(N\u2014H), 3050 (m); \u03bd(C\u2014H), 3081 (s), 3005 (m), 2968 (m), 2938 (m); \u03bd(C\u2261N), 1740 (s), d(N\u2014H) + \u03bd(C\u2261N) + \u03b4(C\u2014H), 1581 (s), 1479 (s), 1401(s); \u03bd(C=S), 1020 (m); \u03bd(P\u2014CPh), 1105 (s); Ligand IR data: \u03bd(N\u2014H), 3130 (s), \u03bd(C\u2014H), 2983 (m); 2876 (s); \u03bd(C\u2261N) 1586 (s), \u03b4(N\u2014H), 1478 (s), 1266 (m); \u03bd(C=S), 1120 (m). The compound is soluble in methanol, chloro\u00adform and partially in di\u00adchloro\u00admethane.To the solution of PdCl6.Uiso(H) = 1.2Ueq(C). The structure of 2 contains partially occupied aceto\u00adnitrile solvent mol\u00adecules with occupancies of 0.33 and 0.25.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989023000166/ex2063sup1.cifCrystal structure: contains datablock(s) 1, 2. DOI: 10.1107/S2056989023000166/ex20631sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989023000166/ex20632sup3.hklStructure factors: contains datablock(s) 2. DOI: 2234599, 2234598CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title charge-neutral complex is a low-spin complex with a moderately distorted pseudo-octa\u00adhedral coordination environment of the metal ion. As a result of their asymmetric shape, the mol\u00adecules stack into chains, which eventually pack into layers and, finally, into a three-dimensional network connected by weak C\u2014H\u22efN, C\u2014H\u22efC hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions. II(C17H12BrN6O)2]\u00b72MeOH, consists of a charge-neutral complex mol\u00adecule and two independent mol\u00adecules of methanol. In the complex mol\u00adecule, the two tridentate ligand mol\u00adecules 2--6-(1H-pyrazol-1-yl)pyridine coordinate to the FeII ion through the N atoms of the pyrazole, pyridine and triazole groups, forming a pseudo-octa\u00adhedral coordination sphere around the central ion. In the crystal, neighbouring asymmetric mol\u00adecules are linked through weak C\u2014H(pz)\u22ef\u03c0(ph) inter\u00adactions into chains, which are then linked into layers by weak C\u2013H\u22efN/C inter\u00adactions. Finally, the layers stack into a three-dimensional network linked by weak inter\u00adlayer C\u2014H\u22ef\u03c0 inter\u00adactions between the meth\u00adoxy groups and the phenyl rings. The inter\u00admolecular contacts were qu\u00adanti\u00adfied using Hirshfeld surface analysis and two-dimensional fingerprint plots, revealing the relative contributions of the contacts to the crystal packing to be H\u22efH 34.2%, H\u22efC/C\u22efH 25.2%, H\u22efBr/Br\u22efH 13.2%, H\u22efN/N\u22efH 12.2% and H\u22efO/O\u22efH 4.0%. The average Fe\u2014N bond distance is 1.949\u2005\u00c5, indicating the low-spin state of the FeII ion. Energy framework analysis at the HF/3\u201321\u2005G theory level was performed to qu\u00adantify the inter\u00adaction energies in the crystal structure.The unit cell of the title compound, [Fe The two independent methanol mol\u00adecules form O\u2014H\u22efN hydrogen bonds with the triazole (trz) rings of the ligand mol\u00adecules Fig.\u00a01. The cends Fig.\u00a01.6 coordination environment , where \u03c6i is the angle N\u2013Fe\u2013N\u2032 , where \u03b8i is the angle generated by superposition of two opposite faces of an octa\u00adhedron but is, however, in the expected range for bis\u00adazole\u00adpyridines and similar ligands (see below). The calculated continuous shape measure (CShM) value relative to the ideal hO symmetry is 2.24 \u2005\u00c5 . Through weak inter\u00admolecular C\u2014H\u22ef N/C inter\u00adactions in the range 3.128\u2005(14)\u20133.734\u2005(11)\u2005\u00c5 . The layers stack with inter\u00adlayer inter\u00adactions limited to C\u2014H\u22efN(trz) and C\u2014H\u22ef\u03c0(ph) contacts involving the methyl groups . The voids between the layers are occupied by methanol mol\u00adecules, which also participate in bonding between neighbouring layers \u22ef\u03c0(ph) inter\u00admolecular contact between the pyrazole (pz) and phenyl (ph) groups respectively Table\u00a01. The mon\u00c5 Table\u00a01, neighbone Fig.\u00a02b,c. Theps Fig.\u00a02c. The v4.CrystalExplorer to 2.4335 (blue) a.u. , the dispersion forces (Edis) and the total energy diagrams (Etot), are shown in Fig.\u00a05Edis), reflecting the dominating inter\u00adactions in the lattice of the neutral asymmetric mol\u00adecules. The topology of the energy framework resembles the topology of the inter\u00adactions within and between the layers described above. The calculated values Etot are in the range 65.2\u201387.6\u2005kJ\u2005mol\u22121 for intra\u00adchain and intra\u00adlayer inter\u00adactions, whereas for the inter\u00adlayer inter\u00adactions they are within 7.7\u201323.4\u2005kJ\u2005mol\u22121. The colour-coded inter\u00adaction mappings within a radius of 3.8\u2005\u00c5 of a central reference mol\u00adecule for the title compound together with full details of the various contributions to the total energy (Etot) are given in the supporting information.The energy framework change correspondingly, and in the low-spin state they are systematically lower than in the high-spin state. Table\u00a02A search of the Cambridge Structural Database ](BF4)2 prepared by dissolving L = 2--6-(1H-pyra\u00adzol-1-yl)pyridine and Fe(BF4)2\u00b76H2O in boiling acetone, to which chloro\u00adform (5\u2005ml) was then added. The middle layer was a methanol\u2013chloro\u00adform mixture , which was covered by a layer of methanol (10\u2005ml), to which 100\u2005\u00b5l of NEt3 was added dropwise. The tube was sealed, and black cubic single crystals appeared in 3\u20134 weeks (yield ca 60%). Elemental analysis calculated for C36H32Br2FeN12O4: C, 47.39; H, 3.54; N, 18.42. Found: C, 47.11; H, 3.74; N, 18.40.The synthesis of the title compound is identical to that reported recently for a similar complex (Seredyuk 8.Uiso(H) = 1.2\u20131.5Ueq(C)]. O-bound H atoms were refined with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022010179/dj2053sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022010179/dj2053Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022010179/dj2053Isup4.cdxSupporting information file. DOI: 10.1107/S2056989022010179/dj2053sup3.pdfIncludes energy framework data and schematic structures of similar neutral Fe(II) complexes. DOI: 2215273CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Mutations can occur in G\u03b1 proteins which prevent GTP hydrolysis; this allows the G proteins to signal independently of G protein\u2013coupled receptors and can result in various cancers, such as uveal melanoma (UM). Most UM cases harbor Q209L, Q209P, or R183C mutations in G\u03b1q/11 proteins, rendering the proteins constitutively active (CA). Although it is generally thought that active, GTP-bound G\u03b1 subunits are dissociated from and signal independently of G\u03b2\u03b3, accumulating evidence indicates that some CA G\u03b1 mutants, such as G\u03b1q/11, retain binding to G\u03b2\u03b3, and this interaction is necessary for signaling. Here, we demonstrate that disrupting the interaction between G\u03b2\u03b3 and G\u03b1q is sufficient to inhibit aberrant signaling driven by CA G\u03b1q. Introduction of the I25A point mutation in the N-terminal \u03b1 helical domain of CA G\u03b1q to inhibit G\u03b2\u03b3 binding, overexpression of the G protein G\u03b1o to sequester G\u03b2\u03b3, and siRNA depletion of G\u03b2 subunits inhibited or abolished CA G\u03b1q signaling to the MAPK and YAP pathways. Moreover, in HEK 293\u00a0cells and in UM cell lines, we show that G\u03b1q-Q209P and G\u03b1q-R183C are more sensitive to the loss of G\u03b2\u03b3 interaction than G\u03b1q-Q209L. Our study challenges the idea that CA G\u03b1q/11 signals independently of G\u03b2\u03b3 and demonstrates differential sensitivity between the G\u03b1q-Q209L, G\u03b1q-Q209P, and G\u03b1q-R183C mutants.Heterotrimeric G protein stimulation Q209L or Q209P mutations in G\u03b1q or G\u03b111 occur in 90% of UM patients, and mutations at R183 occur in about 5% of patients (Heterotrimeric G proteins (G\u03b1\u03b2\u03b3) are canonically activated through G protein\u2013coupled receptors (GPCRs), which allows for GDP release from the G\u03b1 subunit and further GTP binding. It is generally accepted that GTP binding on the G\u03b1 subunit promotes an active conformation and allows for the dissociation from G\u03b2\u03b3. Active GTP-bound G\u03b1 proteins can further bind and stimulate downstream effectors , 2. Hydrc tumors , 6. Mutapatients . Approxipatients .q/11 in UM, driven by the activating Q209L/P and R183 mutations, promotes the stimulation of two major signaling pathways: 1) the mitogen-activated protein kinase (MAPK) pathway and 2) the Yes-associated protein and Transcriptional coactivator with PDZ-binding motif (YAP/TAZ) pathway. The MAPK cascade is stimulated by the direct binding of CA G\u03b1q/11 to phospholipase C\u2013\u03b2 (PLC-\u03b2), which hydrolyzes phosphatidylinositol 4,5-bisphosphate into diacylglycerol and inositol 1,4,5-trisphosphate. Diacylglycerol further activates PKC, which phosphorylates and activates RasGRP3. The further activation of Ras promotes the stimulation of the MAPK cascade and results in the phosphorylation and activation of ERK. Phospho-ERK (pERK) dimerizes and translocates into the nucleus to bind transcription factors and promote the transcription of proliferative genes transcription factor in the nucleus to promote the transcription of genes involved in proliferation and cell survival and FR900359 (FR) have been shown to inhibit both WT and CA G\u03b1q/11 and have shown promising results in cancer cell models , Q209P (QP), and R183C (RC)\u2014is sufficient to inhibit oncogenic signaling of these mutationally active G\u03b1q. We provide evidence that disrupting the interaction between CA G\u03b1q and G\u03b2\u03b3 significantly inhibits G\u03b1q-mediated activation of the MAPK and YAP/TAZ pathways. Surprisingly, we show differential sensitivity between G\u03b1q-QL and the G\u03b1q-QP/RC mutants in that oncogenic signaling by G\u03b1q-QP and G\u03b1q-RC is more sensitive to the disruption of G\u03b2\u03b3 binding than G\u03b1q-QL.Here, we tested the hypothesis that disrupting the interaction between G\u03b2\u03b3 and the CA G\u03b1q, such as PLC-\u03b2, requires the binding of G\u03b1q to G\u03b2\u03b3 , Q209P (QP), and R183C (RC) mutants. To monitor the activity of the YAP/TAZ pathway, we transfected the CA mutants, with and without the I25A mutation, into HEK 293\u00a0G\u03b1q/11 CRISPR-Cas9 KO cells and monitored YAP activity using the TEAD luciferase reporter assay, which is stimulated in response to YAP activation and nuclear translocation. G\u03b1q-QL, G\u03b1q-QP, and G\u03b1q-RC all showed strong activation in the assay. Cells expressing the pcDNA3 vector and WT G\u03b1q were used as negative controls and indicated no significant change in TEAD luciferase reporter activity (A). The G\u03b1q-QL-C9,10S palmitoylation-deficient mutant was used as a negative control for G\u03b1q-QL. The C9,10S mutation in WT G\u03b1q has been previously established to prevent plasma membrane localization and consequently prevent GPCR-dependent activation . It has been previously established that activation of the Rho-dependent YAP pathway results in stabilization and dephosphorylation of YAP and its subsequent translocation into the nucleus as a control resulted in the loss of nuclear YAP and its accumulation in the cytoplasm. G\u03b1q-I25A-QL also promoted nuclear localization of YAP, consistent with its retention of signaling in the TEAD-luciferase assay (A). Cells expressing CA G\u03b1q-QP and G\u03b1q-RC had robust nuclear YAP localization, indicating stimulation of this pathway. However, G\u03b1q-I25A-QP and G\u03b1q-I25A-RC failed to stimulate the nuclear localization of YAP, as indicated by the cytoplasmic localization of YAP. Similar to the TEAD-luciferase reporter assay results, this data suggests that G\u03b1q-I25A-QL is more resistant to disruption of signaling than the G\u03b1q-I25A-QP and G\u03b1q-I25A-RC mutants. In immunofluorescence microscopy studies, we also examined the subcellular localization of the CA G\u03b1q mutants with and without the I25A mutation . Expression of CA G\u03b1q-QL resulted in pERK stimulation >4-fold compared to cells transfected with pcDNA3 or WT G\u03b1q. The G\u03b1q-I25A-QL mutant also activated pERK at a slightly decreased level compared to G\u03b1q-QL, although the difference was not statistically significant. Interestingly, the G\u03b1q-QP and G\u03b1q-RC mutants stimulated pERK, but pERK activity was significantly decreased in cells expressing the G\u03b1q-I25A-QP and G\u03b1q-I25A-RC mutants (C and D). The effects of the I25A mutants on MAPK activity were further validated with the serum response element (SRE) luciferase reporter assay, which monitors both Rho and MAPK-dependent activity (E). As indicated, the CA G\u03b1q-QL mutant robustly stimulated SRE-dependent luciferase activity. Introduction of the I25A mutation into G\u03b1q-QL (G\u03b1q-I25A-QL) strongly inhibited the ability to stimulate SRE luciferase activity, similar to the lack of signaling by the G\u03b1q-QL-C9,10S palmitoylation-deficient mutant and the loss of signaling upon treatment of cells expressing G\u03b1q-QL with YM. As expected, the CA G\u03b1q-QP and G\u03b1q-RC mutants also robustly stimulated SRE luciferase activity, while the G\u03b1q-I25A-QP and G\u03b1q-I25A-RC mutants completely failed to stimulate luciferase activity, similar to cells treated with YM (E). The decreased signaling by G\u03b1q-I25A-QL in comparison to G\u03b1q-QL is much more drastic in the SRE luciferase assay than the pERK immunoblot assay (C and D). Although this is somewhat surprising since the SRE luciferase readout is downstream of the MAPK pathway, we note that the SRE luciferase assay provides a very high signal-to-noise readout for G\u03b1q-QL signaling and is consistently very sensitive to disruption of G\u03b1q-QL signaling . Taken together, these results indicate that the introduction of the I25A mutation to disrupt binding to G\u03b2\u03b3 inhibited oncogenic signaling of the CA G\u03b1q mutants through the YAP and MAPK pathways, and the G\u03b1q-QP and G\u03b1q-RC mutants are more sensitive to the I25A mutation than G\u03b1q-QL.Along with stimulation of the YAP pathway, CA G\u03b1 mutants , C and D mutants , C and Dactivity E. As ind with YM E. The deot assay , C and Dq-QL has an increased association with G\u03b2\u03b3 compared to G\u03b1q-RC and was further quantified (B). As expected, WT G\u03b1q displayed a strong pull down with G\u03b21\u03b32, but the introduction of the I25A mutation into WT G\u03b1q strongly decreased this interaction, indicating that the I25A mutation disrupts binding between G\u03b1q and G\u03b2\u03b3. Interestingly, G\u03b1q-QL had a similar binding association to G\u03b21\u03b32 with that of WT G\u03b1q, and G\u03b1q-QL bound significantly stronger to G\u03b21\u03b32 than G\u03b1q-QP and G\u03b1q-RC (A and B). Furthermore, G\u03b1q-I25A-QL was more strongly bound to G\u03b21\u03b32 than G\u03b1q-I25A-QP and G\u03b1q-I25A-RC (A and B). We did not see a statistically significant difference in association with G\u03b2\u03b3 between G\u03b1q-QP and G\u03b1q-RC and their corresponding I25A mutants. This is likely due to the low initial binding to G\u03b2\u03b3 with both G\u03b1q-QP and G\u03b1q-RC. Although we previously demonstrated the surprising ability of G\u03b1q-QL to interact strongly with G\u03b2\u03b3 and the poor interaction of G\u03b1q-RC with G\u03b2\u03b3, we now show a dramatically decreased association of G\u03b1q-QP with G\u03b2\u03b3 compared to G\u03b1q-QL, even though both have a mutation of Q209. This differential binding to G\u03b2\u03b3 between the CA G\u03b1q mutants likely contributes to the varying sensitivity in oncogenic signal disruption between the I25A mutants . Interestingly, cotransfection with 200\u00a0ng of the G\u03b1o DNA was required to decrease G\u03b1q-QL stimulation of TEAD luciferase activity by 63%. In comparison, cotransfection with 100\u00a0ng of G\u03b1o plasmid DNA achieved 83% and 84% reduction in signaling by G\u03b1q-QP and G\u03b1q-RC, respectively. Transfection with the control G\u03b1o G2A constructs did not reduce the luciferase reporter activity in response to the CA G\u03b1q mutants (A). The ability of the expression of G\u03b1o to reduce CA G\u03b1q-stimulated YAP activity was also monitored through nuclear localization of YAP by immunofluorescence microscopy. G\u03b1o\u00a0localized strongly at cellular membranes, while the G\u03b1o G2A mutant was primarily cytoplasmic, indicating the expected localization of the proteins. Strong nuclear YAP was detected with the CA G\u03b1q-QL, G\u03b1q-QP, and G\u03b1q-RC mutants, indicating stimulation of the YAP pathway (B). As expected, expression of G\u03b1o G2A with the CA G\u03b1q mutants did not disrupt nuclear localization of YAP. Overexpression of G\u03b1o inhibited YAP nuclear localization stimulated by G\u03b1q-QP and G\u03b1q-RC (B). Conversely, cotransfection of G\u03b1o with G\u03b1q-QL failed to prevent strong YAP nuclear localization (B), further suggesting that the G\u03b1q-QP and G\u03b1q-RC mutants are more sensitive to G\u03b2\u03b3 binding disruption than G\u03b1q-QL.Considering that inhibiting the association between the CA G\u03b1in cells , 42, 43.embranes . YAP actplasmids A. Intere mutants A. The ab pathway B. As expd G\u03b1q-RC B. Converlization B, furtheo expression and subsequent G\u03b2\u03b3 sequestration on CA G\u03b1q-promoted MAPK activity, we immunoblotted for pERK levels (A and B). The expression of the CA G\u03b1q mutants resulted in strong stimulation of pERK. Expression of G\u03b1o resulted in no significant decrease in pERK activity for G\u03b1q-QL. However, cotransfection of G\u03b1q-QP or G\u03b1q-RC with 50\u00a0ng and 100\u00a0ng of G\u03b1o plasmid resulted in a strong decrease in pERK activity (A and B). There was no significant change in pERK levels with the G\u03b1o G2A control (A and B). These results were further validated using the SRE luciferase reporter assay (C). Expression of G\u03b1q-QL, G\u03b1q-QP, and G\u03b1q-RC robustly activated SRE-dependent luciferase activity, while treatment with YM abolished luciferase activity. Consistent with results from the other signaling assays (A and B), cotransfection with 50\u00a0ng\u00a0G\u03b1o plasmid was sufficient to completely abolish G\u03b1q-QP\u2013 and G\u03b1q-RC\u2013stimulated SRE luciferase activity, while up to 250\u00a0ng of G\u03b1o plasmid was required for near complete inhibition of G\u03b1q-QL\u2013stimulated SRE luciferase activity (C). These findings indicate that the expression of G\u03b1o to sequester endogenous G\u03b2\u03b3 inhibits the signaling activity of G\u03b1q-QL/P and G\u03b1q-RC. The results also further validate that the G\u03b1q-QL CA mutant is less sensitive to signaling disruption than the G\u03b1q-QP and G\u03b1q-RC mutants.To monitor the effects of G\u03b1K levels , A and Bactivity , A and B control , A and Ber assay C. Expresg assays , A and Bactivity C. These o could sequester G\u03b2\u03b3 and inhibit the binding of G\u03b2\u03b3 to G\u03b1q-QL/P and G\u03b1q-RC. To examine this, we coexpressed G\u03b1o or G\u03b1o G2A with the CA G\u03b1q mutants in 6x-His-G\u03b21y2 HEK 293 stable cells and pulled down G\u03b21\u03b32 with Ni-NTA beads. Immunoblots were used to detect G\u03b1q and G\u03b1o bound to G\u03b21 (A and B). As indicated in q-QL had a stronger binding association with G\u03b21 than G\u03b1q-QP and G\u03b1q-RC. G\u03b1o was bound to G\u03b21 in the pull-down extract, but G\u03b1o G2A had minimal to no interaction with G\u03b21 as expected. The expression of G\u03b1o resulted in significantly decreased binding of G\u03b1q-QL to G\u03b21 (A and B). Expression of G\u03b1o consistently resulted in a noticeable decrease in the pull down of G\u03b1q-QP and G\u03b1q-RC with G\u03b2\u03b3, but this did not reach statistical significance due to the already low level of G\u03b1q-QP and G\u03b1q-RC association with G\u03b2\u03b3 in the absence of G\u03b1o expression. These findings confirm that overexpression of G\u03b1o sequesters G\u03b2\u03b3 and prevents binding of G\u03b1q-QL/P and G\u03b1q-RC to G\u03b2\u03b3.Next, we wanted to confirm that the expression of G\u03b1d to G\u03b21 , A and BL to G\u03b21 , A and Bq through the N-terminal I25A mutation or expression of G\u03b1o is sufficient to inhibit overactive G\u03b1q signaling in HEK 293\u00a0G\u03b1q/11 KO cells. We next wanted to determine if inhibiting G\u03b2\u03b3 binding to CA G\u03b1q would be sufficient to disrupt oncogenic signaling in UM cells containing the G\u03b1q-QL or G\u03b1q-QP mutants. To study this, we depleted G\u03b21 and G\u03b22, two predominant G\u03b2 subunits, through siRNA transfections. Sufficient knockdown of G\u03b21 and G\u03b22 was first validated in HEK 293\u00a0G\u03b1q/11 KO cells (A and B). siRNA molecules specific for G\u03b21, G\u03b22, and both G\u03b21 and G\u03b22 were transfected into the HEK 293\u00a0G\u03b1q/11 KO cells, and WT G\u03b1q, G\u03b1q-QL, or G\u03b1q-QP constructs were transfected after 24\u00a0h. pERK levels were detected and quantified through immunoblotting (A and B). The results indicate that siRNA depletion of G\u03b21/2 significantly disrupted pERK activity driven by both G\u03b1q-QL and G\u03b1q-QP (A and B). G\u03b21 or G\u03b22 siRNA knockdown alone was insufficient to significantly inhibit pERK activation (A and B). Considering the effectiveness of G\u03b21/2 depletion on pERK activity in the HEK 293\u00a0G\u03b1q/11 KO cells, we decided to knock down G\u03b21/2 in four UM cell lines: 1) Mel202 (G\u03b1q-Q209L mutation), 2) 92.1 (G\u03b1q-Q209L mutation), 3) OMM1.3 (G\u03b1q-Q209P mutation), and 4) UM001 (G\u03b1q-Q209P mutation). We also used the OCM-3\u00a0cell line as a control. The oncogenic activity of OCM-3\u00a0cells is driven by mutant BRAF (V600E) as opposed to mutant G\u03b1q/11; thus, we would expect G\u03b21/2 depletion to have minimal effect on pERK activity and quantified (D). Interestingly, the OMM1.3 and UM001\u00a0cell lines which contain the G\u03b1q-Q209P mutation had significantly decreased pERK levels with G\u03b21/2 knockdown, similar to that of YM treatment (C and D). There was no significant difference in pERK levels with G\u03b21/2 depletion in the Mel202 and 92.1\u00a0cells which contain the G\u03b1q-Q209L mutation (C and D). This data further suggests that there is differential sensitivity to the inhibition of oncogenic signaling between cell lines with the G\u03b1q-Q209L and G\u03b1q-Q209P mutations.Our studies demonstrate that disrupting the interaction between G\u03b2\u03b3 and CA G\u03b1KO cells , A and Bblotting , A and Bd G\u03b1q-QP , A and Btivation , A and Bactivity . All celoblotted C and quaantified D. Interereatment , C and Dmutation , C and Dq and show a surprising differential sensitivity of CA G\u03b1q mutants to the inhibition of signaling upon disruption of their interaction with G\u03b2\u03b3. We used two methods to disrupt G\u03b2\u03b3 interaction with CA G\u03b1q\u2014introduction of a G\u03b2\u03b3-binding disrupting mutation, I25A, into CA G\u03b1q and sequestration of endogenous G\u03b2\u03b3 by expression of G\u03b1o\u2014to show that inhibiting interaction with G\u03b2\u03b3 can drastically reduce CA G\u03b1q-stimulated signaling to both the MAPK and YAP/TAZ pathways. Moreover, signaling by CA G\u03b1q-Q209P, as well as CA G\u03b1q-R183C, was much more effectively prevented by G\u03b2\u03b3 binding disruption than signaling by CA G\u03b1q-Q209L. Lastly, we extended the analysis to UM cell lines containing either the oncogenic driver mutant G\u03b1q-Q209L or G\u03b1q-Q209P and showed that depletion of G\u03b21/2 strongly inhibited activation of the MAPK pathway in cells with the G\u03b1q-Q209P mutant but not in cells with the G\u03b1q-Q209L. Our results suggest that disrupting the interaction between G\u03b2\u03b3 and CA G\u03b1q may be a novel approach for inhibiting aberrant G\u03b1q/11 signaling.The results presented in this study demonstrate the importance of G\u03b2\u03b3 for signaling by CA mutants of G\u03b1q mutants studied herein, it would be expected that CA G\u03b1 mutants would not retain binding to G\u03b2\u03b3. However, several studies have challenged the classical heterotrimer dissociation model, indicating that at least some G\u03b1 subunits maintain association with G\u03b2\u03b3 when they are in the active, GTP-bound state due to the presence of a GTPase-inhibiting mutation or in response to GPCR activation a central switch region, which undergoes a conformational change upon GTP binding and 2) an N-terminal \u03b1 helical domain . Thus, one reason why CA G\u03b1q mutants in this study showed decreased or loss of signaling to the MAPK and YAP pathways when interaction with G\u03b2\u03b3 is disrupted is a failure to be efficiently palmitoylated and membrane-bound. Indeed, G\u03b1q-I25A-QL retains stronger plasma membrane localization than G\u03b1q-I25A-QP and G\u03b1q-I25A-RC , although only partial loss of signaling was observed in when measuring pERK (C and D). Expression of the increasing amounts of G\u03b1o to sequester G\u03b2\u03b3 provided a powerful tool to demonstrate titratable differences in sensitivity to disruption of G\u03b2\u03b3 binding to the CA G\u03b1q mutants . Using the SRE-luciferase assay, transfection with 50\u00a0ng of G\u03b1o expression plasmid was sufficient to completely abolish signaling of G\u03b1q-QP and G\u03b1q-RC, while transfection with 250\u00a0ng of G\u03b1o plasmid was required to observe a similar almost-complete inhibition of G\u03b1q-QL signaling (C). Likewise, transfection with 50 or 100\u00a0ng of G\u03b1o plasmid failed to inhibit G\u03b1q-QL\u2013stimulated pERK levels, but transfection with 50 or 100\u00a0ng of G\u03b1o plasmid provided significant inhibition of G\u03b1q-QP\u2013 and G\u03b1q-RC\u2013stimulated pERK (A and B). G\u03b2\u03b3 pull-down experiments showed the interaction of G\u03b1q-QL with G\u03b2\u03b3, similar to the levels of WT G\u03b1q interaction with G\u03b2\u03b3, and, importantly, the I25A mutation and expression of G\u03b1o only partially disrupted the interaction . The G\u03b1q-I25A, G\u03b1q-I25A-QL, and G\u03b1q-I25A-RC constructs were previously described . YFP-tagged G\u03b1q was provided by Catherine Berlot. G\u03b1o and G\u03b1o-G2A plasmids were described previously (q and G\u03b1o transient transfections was obtained from Invitrogen (Cat # 11668-019). siRNA targeting G\u03b21 (GGAUAACAUUUGCUCCAUU), G\u03b22 (ACUGGGUACCUGUCGUGUU), and both G\u03b21 and G\u03b22 (G\u03b21/2) (ACGACGACUUCAACUGCAA) were previously described (for immunoblots with HEK 293\u00a0G\u03b1q/11 KO lysates), GAPDH (Cat # 60004-1-Ig), and G\u03b1o (Cat # 12635-1-AP) were from Proteintech. The G\u03b1q (Cat # ab199533) (for immunofluorescence experiments), G\u03b21 (Cat # ab137635), and G\u03b22 (Cat # ab108504) antibodies were obtained from Abcam. The YAP antibody (Cat # sc-101199) was purchased from Santa Cruz. The G\u03b1q (Cat # 14373S) , ERK (Cat # 4696S), pERK (Cat # 9101S), and Myc-tag (Cat # 2272S) antibodies were obtained from Cell Signaling Technologies. The HA-tag antibody 12CA5 was from Covance. The GRK4-6 antibody (Cat # 05-466) was obtained from Sigma-Aldrich. For immunofluorescence microscopy, the secondary antibodies Alexa Fluor 488 (goat anti-rabbit) (Cat # A-11034), Alexa Fluor 594 (goat anti-mouse) (Cat # A-11032), Alexa Fluor 594 (goat anti-rabbit) (Cat # A-11037), and Alexa Fluor 647 (goat anti-mouse) (Cat # A-32728) were purchased from Invitrogen. The secondary antibodies IRDye 680RD goat anti-rabbit IgG (H\u00a0+ L) (Cat # 92568071) and IRDye 800CW donkey anti-mouse IgG (H\u00a0+ L) (Cat # 92532212) were obtained from LI-COR and were used to visualize protein from all of the immunoblots.The antibodies for G\u03b1q/11 KO cells were generously provided by Dr Asuka Inoue and were described previously with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin . HEK 293\u00a0G\u03b21\u03b32 stable cells were described previously . The SRE luciferase reporter plasmid was described previously with 1% Triton X-100. For the G\u03b1q-I25A experiments, the cells were incubated with the anti-rabbit G\u03b1q antibody (Abcam) and the anti-mouse YAP antibody in 2.5% milk/TBS-Triton X-100. For the G\u03b1o experiments, the cells were incubated with the anti-rabbit G\u03b1o antibody and the anti-mouse YAP antibody in 2.5% milk/TBS-Triton X-100. The primary antibodies were incubated for 60\u00a0min. The cells were washed 5 times with 2.5% milk/TBS-Triton X-100. For the G\u03b1q-I25A experiments, the cells were incubated with goat anti-rabbit Alexa Fluor 488 and goat anti-mouse Alexa Fluor 594 antibodies in 2.5% milk/TBS-Triton X-100. For the G\u03b1o experiments, the cells were incubated with goat anti-rabbit Alexa Fluor 594 and goat anti-mouse Alexa Fluor 647 antibodies in 2.5% milk/TBS-Triton X-100. The secondary antibodies were incubated for 30\u00a0min. The cells were washed 5 times in TBS/1% Triton X-100. The cells were incubated in DAPI diluted in warmed PBS for 5\u00a0min. The coverslips were rinsed in distilled water and mounted onto glass slides with ProLong Diamond Anti-fade Mountant . The images were acquired using the Olympus IX83 microscope with a 60\u00d7 oil immersion objective and an ORCA Fusion sCMOS camera (Hamamatsu) controlled by Olympus cellSens software.HEK 293\u00a0G\u03b11\u03b32 pull-down assay was done as previously described . The cell lysates were incubated for 1\u00a0h on ice and were then centrifuged at 13,000\u00a0rpm to pellet the nuclei and insoluble material. Forty microliters of the lysate were reserved separately for the input fraction. The remaining lysate was added to 30\u00a0\u03bcl of Ni-NTA beads and rotated in an end-over-end rotator for 2\u00a0h at 4 \u00b0C. The tubes were placed in a magnetic rack, and the remaining supernatant was aspirated. The beads were washed 3 times with lysis buffer C. Fifty microliters of elution buffer (lysis buffer C with 0.25\u00a0M Imidazole) were added to elute G\u03b21 and any bound proteins from the Ni-NTA beads. Forty microliters of the pull-down eluate were transferred to a new tube. Ten microliters of 5\u00d7 SDS-PAGE sample buffer with 3.5% \u03b2-mercaptoethanol were added to the input fraction and the pull-down fraction. The input and pull-down lysates were separated on a 10% SDS-PAGE gel and protein bound was detected via immunoblotting. The blots were probed with the HA-tag and Myc-tag antibody . The blots for the G\u03b1o experiments were also probed with G\u03b1o and GAPDH. The blots were imaged on LI-COR, and the bands were quantified on the ImageJ software (https://imagej.nih.gov/ij/). The pull-down samples in G\u03b1q-I25A experiments were normalized to its respective input fraction. The pull-down samples in the G\u03b1o experiments were normalized to WT G\u03b1q.The G\u03b2escribed . Brieflyq for the HEK 293\u00a0G\u03b1q/11 KO cells or Cell Signaling G\u03b1q for the UM cells, along with GAPDH as a loading control. The ERK and pERK bands were quantified by densitometry using the ImageJ software, and the relative pERK signal was divided by ERK. The quantified signals were normalized to WT G\u03b1q (D), the individual CA G\u03b1q mutants (B), or control siRNA between the CA G\u03b1q mutants (B) or the UM cell lines (D).Cells were lysed in SDS-PAGE sample buffer. Lysates were run on a 10% SDS-PAGE gel and transferred to a nitrocellulose membrane. The membrane was blocked in 2.5% bovine serum albumin (BSA) in 1\u00d7 TBS supplemented with 0.05% Tween 20. The blots were incubated with both pERK and ERK antibodies overnight. A duplicate immunoblot was performed and was probed with either Proteintech G\u03b1o WT G\u03b1q D, the in mutants B, or con mutants B or the ll lines D.q/11 KO cells and 12-well plates for the UM cells. For the HEK 293\u00a0G\u03b1q/11 KO experiments, 30 pmol of control, G\u03b21, G\u03b22, or G\u03b21/2 siRNA were transfected into cells using Lipofectamine RNAiMax , according to manufacturer's instructions. The media was changed after 5\u00a0h. After 24\u00a0h, the corresponding WT G\u03b1q, G\u03b1q-QL, or G\u03b1q-QP constructs were transfected into the cells using Lipofectamine 2000 . The media was changed to serum-free media after 24\u00a0h and corresponding cells were treated with 1\u00a0\u03bcM of YM. The cells were lysed in 1\u00d7 SDS-PAGE sample buffer with 0.7% \u03b2-mercaptoethanol after 16\u00a0h. For experiments with the UM cells, 15pmol of control or G\u03b21/2 siRNA were transfected into cells using Lipofectamine RNAiMax . The media was changed after 5\u00a0h. After 72\u00a0h, the media was changed to serum-free media, and the corresponding cells were treated with 1\u00a0\u03bcM of YM. The cells were further lysed after approximately 16\u00a0h in 1\u00d7 SDS-PAGE sample buffer with 0.7% \u03b2-mercaptoethanol. The lysates were run on 10% SDS-PAGE gels and were immunoblotted for pERK , ERK , G\u03b21 , G\u03b22 , GAPDH , and Proteintech G\u03b1q (Cat #13927-1-AP) for the HEK 293\u00a0G\u03b1q/11 KO cells or Cell Signaling G\u03b1q (Cat # 14373S) for the UM cells.Cells were plated in 6-well plates for the HEK 293\u00a0G\u03b1Protein lysates were run on 10% SDS-PAGE gels and transferred to LI-COR nitrocellulose membranes (Cat # nc9680617). The membranes were blocked in either 2.5% BSA or 2.5% milk in 1\u00d7 TBS/0.05% Tween 20 at room temperature for 60\u00a0min. The blots were incubated in corresponding primary antibodies in 2.5% BSA or milk in 1\u00d7 TBS/0.05% Tween 20 at 4 \u00b0C overnight. The blots were washed three times in 1\u00d7 TBS/0.05% Tween 20 and incubated in LI-COR anti-rabbit (Cat # 92568071) and anti-mouse (Cat # 92532212) secondary antibodies for 60\u00a0min at room temperature. The immunoblots were further washed three times in 1\u00d7 TBS/0.05% Tween 20 and once with PBS. The blots were then imaged on the LI-COR Odyssey imager.B and D) multiple comparison test were used to calculate significance. Error bars in all experiments indicate mean\u00a0\u00b1 SD with significant differences indicated as \u2217p\u00a0< 0.05; \u2217\u2217p\u00a0< 0.01; \u2217\u2217\u2217p\u00a0< 0.005; \u2217\u2217\u2217\u2217p\u00a0< 0.0001.GraphPad Prism was used to analyze the data for all of the figures. A two-way ANOVA followed by \u0160id\u00e1k's or Tukey\u2019s , B and DAll data is contained within the article.This article contains The authors declare that they have no conflicts of interest with the contents of this article."} +{"text": "The synthesis and crystal structure of a new Schiff base, namely, 4-bromo-2-[({2-[(2-hy\u00addroxy\u00adeth\u00adyl)amino]\u00adeth\u00adyl}imino)\u00admeth\u00adyl]phenol, is reported. 11H15BrN2O2, crystallizes in the monoclinic space group P21 with two independent mol\u00adecules in the asymmetric unit. It was prepared by the condensation reaction of 5-bromo-2-hy\u00addroxy\u00adbenzaldehyde and amino\u00adethyl\u00adethano\u00adlamine. There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond with an S(6) ring motif. Moreover, there are inter\u00admolecular C\u2014H\u22efN, C\u2014H\u22efO and Br\u22efO inter\u00adactions in the crystal structure of the title compound.The new title Schiff base compound, C Moreover, the O2a and O2b atoms are involved in a second intra\u00admolecular hydrogen bond. The mol\u00adecules are connected through inter\u00admolecular O\u2014H\u22efN hydrogen bonds and Br\u22efO inter\u00adactions with distances Br1a\u22efO2b = 3.206\u2005(2)\u2005\u00c5 and Br1b\u22efO2a = 3.282\u2005(2)\u2005\u00c5 was dissolved in ethanol (10\u2005ml) and stirred for 10\u2005min. Then, a solution of amino\u00adethyl\u00adethano\u00adlamine (0.2\u2005mmol) dissolved in ethanol (5\u2005ml) was added dropwise. The mixture was stirred and refluxed for 6\u2005h. After that, the solution was concentrated under reduced pressure. Yellow crystals suitable for X-ray analysis were obtained by slow evaporation of solvent at room temperature for several days. These were filtered off and washed several times with cold ethanol.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621003357/bt4109sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314621003357/bt4109Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314621003357/bt4109Isup3.cmlSupporting information file. DOI: 2074082CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II ion of the complex cation has an N3O square-planar coordination sphere defined by the three N atoms of the tridentate 2,2\u2032:6\u2032,2\u2032\u2032-terpyridine ligand and one O atom from the NO3\u2212 anion.The central Pd 3)(C15H11N3)]NO3, comprises a cationic PdII complex and a nitrate anion. In the complex, the PdII cation is four-coordinated in a distorted square-planar coordination geometry defined by the three N atoms of the tridentate 2,2\u2032:6\u2032,2\u2032\u2032-terpyridine ligand and one O atom from the NO3\u2212 anion. In the crystal, the complex mol\u00adecules are stacked in columns along the a axis being connected by \u03c0\u2013\u03c0 stacking [closest inter-centroid separation between pyridyl rings = 3.878\u2005(3)\u2005\u00c5]. The connections between columns and anions to sustain a three-dimensional architecture are C\u2014H\u22efO hydrogen bonds.The title complex, [Pd(NO The Pd\u2014N [1.917\u2005(4) to 2.030\u2005(4)\u2005\u00c5] and Pd\u2014O [2.028\u2005(3)\u2005\u00c5] bond lengths are close. The pyridine rings of the terpy ligand are located approximately parallel to the least-squares plane of the PdN3O unit [maximum deviation = 0.023\u2005(2)\u2005\u00c5], with dihedral angles of 1.4\u2005(2)\u00b0 (ring N1/C1\u2013C5), 3.1\u2005(2)\u00b0 (ring N2/C6\u2013C10) and 3.0\u2005(2)\u00b0 (ring N3/C11\u2013C15). In the crystal and Cg2i , the centroid-centroid distance is 3.878\u2005(3)\u2005\u00c5 and the dihedral angle between the ring planes is 3.2\u2005(3)\u00b0 2\u00b72H2O in acetone (30\u2005ml) was added 2,2\u2032:6\u2032,2\u2032\u2032-terpyridine followed by stirring for 3\u2005h at room temperature. The formed precipitate was separated by filtration, washed with acetone and dried at 323\u2005K to give a light-yellow powder (0.2123\u2005g). Yellow crystals of the product suitable for X-ray analysis were obtained by slow evaporation of its CH3NO2 solution at room temperature.To a solution of Pd I. DOI: 10.1107/S2414314621000857/tk4067Isup2.hklStructure factors: contains datablock(s) I. DOI: 2058389CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "P-substituent benzoate anions or a tri\u00adfluoro\u00adacetate anion. They are hydrated and there are three crystallized as 1:1 salts while the fourth is a 2:2 salt. Their crystal packing depends on strong ribbons or sheets stabilized by hydrogen bonds of type N\u2014H\u22efO and O\u2014H\u22efO and other inter\u00adactions as C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and C\u2014H\u22efF in the tri\u00adfluoro\u00adacetate-based one.Four novel piperazinium salts are reported, and based on 1-phenyl\u00adpiperazinium as a common cation in the asymmetric units that include additionally water mol\u00adecules, and different 9H9O3\u00b7C10H15N2\u00b7H2O (I); 4-phenyl\u00adpiperazin-1-ium 4-meth\u00adoxy\u00adbenzoate monohydrate, C10H15N2\u00b7C8H7O3\u00b7H2O (II); 4-phenyl\u00adpiperazin-1-ium 4-methyl\u00adbenzoate monohydrate, C10H15N2\u00b7C8H7O2\u00b7H2O (III); and 4-phenyl\u00adpiperazin-1-ium tri\u00adfluoro\u00adacetate 0.12 hydrate, C10H15N2\u00b7C2F3O2\u00b70.12H2O (IV), have been synthesized. The single-crystal structures of these compounds reveal that all of them crystallize in the triclinic PI)\u2013(III) is built up of ribbons formed by a combination of hydrogen bonds of type N\u2014H\u22efO, O\u2014H\u22efO and other weak inter\u00adactions of type C\u2014H\u22efO and C\u2014H\u22ef\u03c0, leading to a three-dimensional network. In the crystal of (IV), the cations and the anions are connected by C\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds and by C\u2014H\u22ef\u03c0 inter\u00adactions, forming sheets which in turn inter\u00adact to maintain the crystal structure by linking through the oxygen atoms of water mol\u00adecules and van der Waals inter\u00adactions, giving the whole structure.In this study, four new piperazinium salts, namely, 4-phenyl\u00adpiperazin-1-ium 4-eth\u00adoxy\u00adbenzoate monohydrate, C In compound (II) the asymmetric unit \u2005\u00c5, \u03b8 = 175.89\u2005(17)\u00b0 and \u03c6 = 346\u2005(3)\u00b0. Compound (III) presents an asymmetric unit \u2005\u00c5, \u03b8 = 9.38\u2005(19)\u00b0 and \u03c6 = 167.9\u2005(13)\u00b0. On the other hand, the asymmetric unit of (IV) and two tri\u00adfluoro\u00adacetate anions and a 0.12 occupancy water molecule. The aromatic rings of the cations are essentially planar while the protonated piperazine rings adopt a chair conformation for cation A1, with puckering parameters \u2005\u00c5, \u03b8 = 0.0\u2005(4)\u00b0 and \u03c6 = 207\u2005(14)\u00b0, and a distorted chair conformation for the cation A2, with puckering parameters QT = 0.559\u2005(5)\u2005\u00c5, \u03b8 = 6.6\u2005(4)\u00b0 and \u03c6 = 168\u2005(4)\u00b0.The asymmetric unit of the compound , Fig.\u00a01, consist), Fig.\u00a01 QT = 0.it Fig.\u00a02 compriseit Fig.\u00a03 composedV) Fig.\u00a04 contains3.I), the cation pairs are connected across two water mol\u00adecules by C\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming an a-axis direction (Table\u00a01a). In addition, a set of C\u2014H\u22ef\u03c0 inter\u00adactions, through the benzene rings of the anions and the cations, connect the mol\u00adecules together in ribbons along the a-axis direction (Table\u00a01b). The C\u2014H\u22efO, N\u2014H\u22efO, O\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions together form a three-dimensional network, contributing to the stabilization of the crystal structure.In the crystal structure of (n Table\u00a01a. In adn Table\u00a01b. The CII), the cations, the anions and the water mol\u00adecules are connected by C\u2014H\u22efO, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, forming ribbons along the a-axis direction (Table\u00a02a). Furthermore, the cations inter\u00adact via C\u2014-H\u22ef\u03c0 inter\u00adactions through the benzene ring of the anion, forming ribbons along the b-axis direction (Table\u00a02b). The C\u2014H\u22efO, N\u2014H\u22efO, O\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions together form a three-dimensional network, contributing to the stabilization of the crystal structure.In the crystal structure of (n Table\u00a02a. Furthn Table\u00a02b. The CIII), the cations, the anions and the water mol\u00adecules are connected by C\u2014H\u22efO, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, forming ribbons along the a-axis direction , the cations and the anions are connected by C\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds (Table\u00a04a) and C\u2014H\u22ef\u03c0 inter\u00adactions, generating sheets parallel to the (100) plane (Table\u00a04IV) involve the oxygen atoms of carboxylate groups, while the 0.12 fraction of the water molecule contributes with one interaction of the type C\u2014H\u22efO and it is weak in comparison to the other oxygen-based ones.In the crystal structure of \u2013(III) gave no hits. However, searching for a branched phenyl piperazinium cation and para-substituted benzoate anion gave comparable hits, namely; 4-(4-meth\u00adoxy\u00adphen\u00adyl)piperazin-1-ium 4-fluoro\u00adbenzoate monohydrate, 4-(4-meth\u00adoxy\u00adphen\u00adyl)piperazin-1-ium 4-chloro\u00adbenzoate monohydrate, 4-(4-meth\u00adoxy\u00adphen\u00adyl)piperazin-1-ium 4-bromo\u00adbenzoate monohydrate \u2013(IV) have no substituent. They also crystallize as monohydrates, and their crystal structures are based on differently sized chains of rings formed via a combination of hydrogen bonds of the type N\u2013H\u22efO and O\u2013H\u22efO and other weak inter\u00adactions of types C\u2014H\u22efO and C\u2014H\u22ef\u03c0 to form sheets. In 4-(4-meth\u00adoxy\u00adphen\u00adyl)piperazin-1-ium 4-amino\u00adbenzoate monohydrate gave no hits.A search of the Cambridge Structural Database \u2013(IV), a solution of commercially available (from Sigma\u2013Aldrich) 1-phenyl\u00adpiperazine in methanol (10\u2005ml) was mixed with equimolar solutions of the appropriate organic acids in methanol (10\u2005ml) viz., 4-eth\u00adoxy\u00adbenzoic acid for (I), 4-meth\u00adoxy\u00adbenzoic acid for (II), 4-methyl\u00adbenzoic acid for (III) and tri\u00adfluoro\u00adacetic acid for (IV). The corresponding solutions were stirred for 15\u2005min at room temperature and allowed to stand at the same temperature. X-ray quality crystals were formed on slow evaporation in a week for all compounds, where ethanol:ethyl\u00adacetate (1:1) was used for crystallization. The corresponding melting points were 353\u2013355\u2005K for (I), 368\u2013370\u2005K for (II), 338\u2013340\u2005K for (III) and 385\u2013387\u2005K for (IV).For the synthesis of salts (6.Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C). For the H atoms bonded to the N and O atoms, the atomic coordinates were refined with Uiso(H) = 1.2Ueq(N) and 1.5Ueq(O), . In (IV), the atoms of the CF3 groups of two tri\u00adfluoro\u00adacetate anions are disordered over two sets of sites with site occupancies of 0.737\u2005(3) and 0.263\u2005(3). The corresponding bond distances in the disordered groups were restrained to be equal. The Uij components of these atoms were restrained to be equal and were restrained to approximate isotropic behaviour. The OW1 water molecule was refined with a resulting occupation factor of 0.245\u2005(10) and the H atoms of the water molecule were placed geometrically.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989022006004/dj2048sup1.cifCrystal structure: contains datablock(s) global, I, II, III, IV. DOI: 10.1107/S2056989022006004/dj2048Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022006004/dj2048Isup6.cmlSupporting information file. DOI: 10.1107/S2056989022006004/dj2048IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989022006004/dj2048IIsup7.cmlSupporting information file. DOI: 10.1107/S2056989022006004/dj2048IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989022006004/dj2048IIIsup8.cmlSupporting information file. DOI: 10.1107/S2056989022006004/dj2048IVsup5.hklStructure factors: contains datablock(s) IV. DOI: Click here for additional data file.10.1107/S2056989022006004/dj2048IVsup9.cmlSupporting information file. DOI: 2177037, 2177036, 2177035, 2177034CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A new organic salt of diclofenac, [2-phen\u00adyl]acetate, with mono\u00adethano\u00adlammonium and water has been obtained, and its structure has been established by X-ray analysis. 2H8NO+\u00b7C14H10Cl2NO2\u2212\u00b7H2O, the asymmetric unit contains one cation, one anion and a water mol\u00adecule, all in general positions. A complex network of hydrogen bonds is present in the crystal structure.In the solid-state structure of the title compound derived from diclofenac, C The dicb-axis direction, due to the crystallographic twofold screw axis, via N2\u2014H2B\u22efO1Wii hydrogen bond [2.947\u2005(4)\u2005\u00c5, symmetry code: (ii) \u2212x, y\u00a0\u2212\u00a0z\u00a0+\u00a0c [O1W\u22efO1iv, symmetry code: (iv) x, \u2212y\u00a0+\u00a0z\u00a0+\u00a0i, symmetry code: (i) \u2212x, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01, 2.811\u2005(4)\u2005\u00c5, Fig.\u00a02X\u22efCg \u03c0\u2013ring inter\u00adactions, for C3\u2014H3 and C7\u2014Cl1 bonds, for which the X\u22efCg separations and \u03b3 angles range from 3.533 to 3.958\u2005\u00c5 and from 25.03 to 28.79\u00b0.The ionic form of the title compound serves as a building block for the supra\u00admolecular architecture. In the crystal, the building blocks form screw-like chains along the Crystal Explorer 17.5 at 298\u2005K for 5\u2005min. The solution was then placed in a loosely closed bottle and kept at 298\u2005K for 10 days. The precipitated prismatic crystals were selected for the single-crystal X-ray diffraction analysis.To a solution of 0.1\u2005g (0.52\u2005mmol) of D in 4\u2005ml of ethanol, 32\u2005\u00b5Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314622004412/bh4068sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622004412/bh4068Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622004412/bh4068Isup3.cmlSupporting information file. DOI: 2168795CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I atom of the cationic complex of the title compound, [Ir(C8H12)(C18H15P)(C6H11N3)][BF4] \u00b70.8CH2Cl2, exhibits a distorted square-planar coordination environment.The central Ir 8H12)(C18H15P)(C6H11N3)]BF4\u00b70.8CH2Cl2, has been synthesized and structurally characterized. The central IrI atom of the cationic complex has a distorted square-planar coordination environment, formed by a bidentate cyclo\u00adocta-1,5-diene (COD) ligand, an N-heterocyclic carbene, and a tri\u00adphenyl\u00adphosphane ligand. The crystal structure comprises C\u2014H\u22ef\u03c0(ring) inter\u00adactions that orient the phenyl rings; non-classical hydrogen-bonding inter\u00adactions between the cationic complex and the tetra\u00adfluorido\u00adborate anion are also present. The complex crystallizes in a triclinic unit cell with two structural units and an incorporation of di\u00adchloro\u00admethane solvate mol\u00adecules with an occupancy of 0.8.A new triazole-based N-heterocyclic carbene iridium(I) cationic complex with a tetra\u00adfluorido\u00adborate counter-anion, [Ir(C The distorted square-planar environment around the IrI atom is defined by the bidentate cyclo\u00adocta-1,5-diene (COD) ligand, the carbene C1 atom of the triazole NHC ligand, and the P atom of the tri\u00adphenyl\u00adphosphane ligand. The P1\u2014Ir1\u2014C1 bond angle is 93.88\u2005(10)\u00b0. The N1\u2014C1\u2014N3 bond angle of the coordinating carbene atom significantly differs with a value of 103.3\u2005(3)\u00b0 from the expected 2sp hybridization.The mol\u00adecular structure of the title complex 4]\u2212 group. Additionally, there are non-classical inter\u00admolecular hydrogen-bonding inter\u00adactions between the hydrogen atom of a phenyl group (H10) and a nitro\u00adgen atom of the NHC ligand (N2). Non-classical hydrogen bonding inter\u00adactions are shown as dotted green lines in Fig.\u00a02The crystal packing of the title compound is displayed in Fig.\u00a02A) and a phenyl phosphane ring (C19\u2013C24). This intra\u00admolecular inter\u00adaction displays an H\u22efcentroid distance of 2.61\u2005\u00c5 and a C\u2014H\u22efcentroid angle of 168\u00b0. The inter\u00admolecular C\u2014H\u22ef\u03c0(ring) inter\u00adaction orients phenyl phosphane rings of adjacent moieties as it occurs between a hydrogen atom of a phenyl ring (H21) and an adjacent phenyl ring (C13\u2013C18). The inter\u00admolecular C\u2014H\u22ef\u03c0(ring) inter\u00adaction has an H\u22efcentroid distance of 2.73\u2005\u00c5 and a C\u2014H\u22efcentroid angle of 157\u00b0. The C\u2014H\u22ef\u03c0(ring) inter\u00adactions orient phenyl rings on adjacent moieties (C13\u2013C18 and C19\u2013C24) into an approximately perpendicular arrangement, shown in Fig.\u00a03Both inter\u00admolecular and intra\u00admolecular C\u2014H\u22ef\u03c0(ring) inter\u00adactions are observed and shown as dashed orange lines in Figs. 2 chloro\u00adiridium (1) was synthesized by a previously published procedure Varian spectrometer and referenced to the residual solvent peak of CDCl3 (\u03b4 in p.p.m.).(tri\u00adphenyl\u00adphosphane)iridium(I) tetra\u00adfluorido\u00adborate (2): Tri\u00adphenyl\u00adphosphane and AgBF4 (0.048\u2005g 0.245\u2005mmol) were added to an oven-dried flask containing complex (1) in 10\u2005ml of CH2Cl2, and stirred under N2 in the dark for 90\u2005min. The mixture was filtered through Celite and the solvent was removed under reduced pressure. The bright orange\u2013red solid was washed with pentane and dried under vacuum yielding 0.165\u2005g (86.9%) of the title compound 2. 1H NMR: \u03b4 (p.p.m.) 8.18 , 7.49\u20137.32 , 5.36 2), 4.38, 3.99 , 4.05 , 2.27\u20131.6 , 1.56 . 13C NMR: \u03b4 177.74 (Ir\u2014C), 140.32 (N\u2014CH\u2014N), 132.46\u2013128.38 (Carom), 87.82, 87.43, 85.34, 85.01 (CH of COD), 53.23 [CH(CH3)2], 41.31 (N\u2014CH3), 33.41, 33.18, 31.45, 30.39 CH2 of COD, 24.37, 22.15 [CH(CH3)2]. 31P: \u03b4 17.23.2 was crystallized by slow diffusion of pentane into a CH2Cl2 solution.The title compound Crystal data, data collection, and structure refinement details are summarized in Table\u00a0210.1107/S2414314623000640/wm4181sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314623000640/wm4181Isup2.hklStructure factors: contains datablock(s) I. DOI: 2237810CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The syntheses and low-temperature crystal structures of four organic salts of 4-(4-nitro\u00adphen\u00adyl)piperazine are presented. 10H14N3O2+\u00b7C4H5O4\u2212 (I), 4-(4-nitro\u00adphen\u00adyl)piperazinium 4-amino\u00adbenzoate monohydrate, C10H14N3O2+\u00b7C7H6NO2\u2212\u00b7H2O (II), 4-(4-nitro\u00adphen\u00adyl)piperazinium 2-(4-chloro\u00adphen\u00adyl)acetate, C10H14N3O2+\u00b7C8H6ClO2\u2212 (III) and 4-(4-nitro\u00adphen\u00adyl)piperazinium 2,3,4,5,6-penta\u00adfluoro\u00adbenzoate, C10H14N3O2+\u00b7C7F5O2\u2212 (IV). The salts form from mixtures of N-(4-nitro\u00adphen\u00adyl)piperazine and the corresponding acid in mixed solvents of methanol and ethyl acetate. Salts I, III, and IV are anhydrous, whereas II is a monohydrate. In each structure, the overall conformation of the cation is determined by the disposition of the exocyclic N\u2014C bond of the piperazine ring and twists about the N\u2014C bond between the piperazine ring and its attached 4-nitro\u00adphenyl ring. The packing motifs in each structure are quite different, though all are dominated by strong N\u2014H\u22efO hydrogen bonds, which are augmented in I and II by O\u2014H\u22efO hydrogen bonds, and in III by a \u03c0\u2013\u03c0 stacking inter\u00adaction between inversion-related 4-nitro\u00adphenyl groups.The syntheses and crystal structures are presented for four organic salts of the 4-(4-nitro\u00adphen\u00adyl)piperazinium cation, namely, 4-(4-nitro\u00adphen\u00adyl)piperazinium hydrogen succinate, C The relative twist about these N2\u2014C5 bonds, e.g. qu\u00adanti\u00adfied by the C2\u2014N2\u2014C5\u2014C6 torsional angles determine the overall cation shape. In each case, the 4-nitro group is essentially coplanar with its attached phenyl ring.The overall conformations of the 4-nitro\u00adphenyl\u00adpiperazinium cations in s Figs. 1 \u25b8 \u25b8. WI has minor twists about its three C\u2014C bonds , which leads to a dihedral angle of 34.63\u2005(9)\u00b0 between its carboxyl\u00adate/carb\u00adoxy\u00adlic acid groups. The 4-amino\u00adbenzoate anion of II is close to planar, having a dihedral angle between the carboxyl\u00adate group and its benzene ring of 10.70\u2005(7)\u00b0. The amine group at N4 is also slightly non-planar [the sum of angles about N4 is 349\u2005(2)\u00b0]. In the 2-(4-chloro\u00adphen\u00adyl)acetate anion of III, twists about the C11\u2014C12 and C12\u2014C13 bonds place the carboxyl\u00adate group almost perpendicular [85.02\u2005(9)\u00b0] to the benzene ring. Lastly, in the penta\u00adfluoro\u00adbenzoate anion of IV, the carboxyl\u00adate group is 55.95\u2005(10)\u00b0 out of coplanarity with the phenyl ring.The succinate anion in I, which will be described in more detail in the next section (Supra\u00admolecular features).Throughout all four structures, individual bond lengths and angles take on normal values except for an elongated O\u2014H bond [1.17\u2005(2)\u2005\u00c5] in 3.2 group and the anionic carboxyl\u00adate groups. In I, II, and IV, these hydrogen bonds to the anion are equatorial relative to the piperazine ring, while that in III is axial. Nevertheless, in each structure, the NH2+ group acts as a hydrogen-bond donor through both its hydrogen atoms. In I, III, and IV this is to a second anion, whereas in II it is to the included water mol\u00adecule. Throughout the four structures, all conventional N\u2014H\u22efO and all but one O\u2014H\u22efO hydrogen bonds take on normal distances and angles hydrogen bond [O\u22efO = 2.4367\u2005(10)\u2005\u00c5], which links adjacent hydrogen-succinate anions into chains that propagate parallel to the b-axis direction anions and as an acceptor for an N\u2014H\u22efO hydrogen bond, generates a double-layer network lying parallel to (011) Fig.\u00a07. Of the III joins two pairs of inversion-related ammonium cations and carboxyl\u00adate anions, forming an III also includes the only \u03c0\u2013\u03c0 inter\u00adactions of the four structures, which occurs between inversion-related nitro\u00adphenyl rings, giving an inter\u00adplanar spacing of 3.3352\u2005(15)\u2005\u00c5, though the offset (\u22431.92\u2005\u00c5) is large, leading to a centroid\u2013centroid distance of 3.8495\u2005(15)\u2005\u00c5 piperazine as a guest mol\u00adecule for I, 4-amino\u00adbenzoic acid for II, 2-(4-chloro\u00adphen\u00adyl)acetic acid for III, and 2,3,4,5,6-penta\u00adfluoro\u00adbenzoic acid for IV. The resulting solutions were stirred for 30 minutes at 333\u2005K and allowed to stand at room temperature. X-ray quality crystals formed on slow evaporation of solutions in ethanol:aceto\u00adnitrile (1:1) over the course of a week for all four compounds. The melting points are 398\u2013400\u2005K (I), 473\u2013475\u2005K (II), 431\u2013435\u2005K (III) and 411\u2013415\u2005K (IV).A solution of commercially available (Sigma-Aldrich) 4-nitro\u00adphenyl\u00adpiperazine in methanol (10\u2005ml) was mixed with equimolar solutions of the appropriate acid in methanol (10\u2005ml) and ethyl acetate (10\u2005ml) 6.sp2\u2014H) and 0.99\u2005\u00c5 (R2CH2), using Uiso(H) values constrained to 1.2Ueq of the attached carbon atom. All N\u2014H and O\u2014H hydrogen atoms were refined freely (both coordinates and Uiso).Crystal data, data collection, and structure refinement details are given in Table\u00a0510.1107/S2056989023001093/hb8053sup1.cifCrystal structure: contains datablock(s) I, II, III, IV, global. DOI: 10.1107/S2056989023001093/hb8053Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989023001093/hb8053IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989023001093/hb8053IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 10.1107/S2056989023001093/hb8053IVsup5.hklStructure factors: contains datablock(s) IV. DOI: Click here for additional data file.10.1107/S2056989023001093/hb8053Isup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001093/hb8053IIsup7.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001093/hb8053IIIsup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001093/hb8053IVsup9.cmlSupporting information file. DOI: 2239906, 2239905, 2239904, 2239903CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Correction: BMC Musculoskelet Disord 23, 630 (2022)https://doi.org/10.1186/s12891-022-05517-0Following the publication of the original article the auth\u201cTheir mean age at surgery was 60.8\u2009\u00b1\u200910.3 years and the average symptom duration was 40.7\u2009\u00b1\u200934.1 months.\u201dShould be corrected to:\u201cTheir mean age at surgery was 61.2\u2009\u00b1\u200910.8 years and the average symptom duration was 23.36\u2009\u00b1\u200922.11 months.\u201dThe original article has been"} +{"text": "II(C68H44N12O4)(C4H6N2)2]\u00b7C4H6N2\u00b71.5C4H8O is investigated.The crystal structure of the bis\u00ad(1-methyl\u00adimidazole)-ligated iron(II) picket-fence porphyrin derivative [Fe II(C68H44N12O4)(C4H6N2)2]\u00b7C4H6N2\u00b71.5C4H8O, the central FeII ion is coordinated by four pyrrole N atoms of the porphyrin core and two N atoms of the 1-methyl\u00adimidazole ligands in the axial sites. One 1-methyl\u00adimidazole and one and a half tetra\u00adhydro\u00adfuran solvent mol\u00adecules are also present in the asymmetric unit. The complex exhibits a near planar porphyrin core conformation, in which the iron centre is slightly displaced towards the hindered porphyrin side (0.01\u2005\u00c5). The average Fe\u2014Np (Np refers to the pyrrole nitro\u00adgen atoms in the porphyrin) bond length is 1.990\u2005(9)\u2005\u00c5, and the axial Fe\u2014NIm (NIm refers to the imidazole nitro\u00adgen atoms) bond lengths are 1.993\u2005(3) and 2.004\u2005(3)\u2005\u00c5. The dihedral angle between the two coordinated 1-methyl\u00adimidazole planes is 56.6\u2005(2)\u00b0. The dihedral angles between the 1-methyl\u00adimidazole planes and the planes of the closest Fe\u2014Np vector are 16.8\u2005(2) and 39.8\u2005(2)\u00b0. N\u2014H\u22efN and N\u2014H\u22efO inter\u00adactions are observed in the crystal structure.In the title compound, [Fe Additional qu\u00adanti\u00adtative information on the structure is given in Fig.\u00a02p vector are 16.8\u2005(2) and 39.8\u2005(2)\u00b0. The dihedral angle between the two coordinated imidazole planes is 56.6\u2005(2)\u00b0, showing a relative perpendicular orientation. Fig.\u00a02p\u2014Fe\u2014Np angle is ideal at 90.01\u2005(9)\u00b0, and the axial Fe\u2014NIm bond lengths are 1.993\u2005(3) and 2.004\u2005(3)\u2005\u00c5. The average Fe\u2014Np distance of 1.990\u2005(9)\u2005\u00c5 is a typical value for low-spin ferrous porphyrin derivatives was distilled from Na/benzo\u00adphenone under N2. Hexanes were distilled over sodium/potassium alloy under N2. Solvents were degassed by repeated freeze\u2013pump\u2013thaw cycles. 1-MeIm was distilled under argon before use. Precursors H2TPyPP, [Fe(TPyPP)]Cl, and [Fe(TPyPP)]OH were prepared following literature methods , with slight modifications.Synthesis of the title compound. [Fe(TPyPP)]OH and 1-MeIm were dissolved in 3\u2005ml of THF. The mixture was stirred for 15\u2005min and transferred into glass tubes (8\u2005mm \u00d7 10\u2005cm), which were layered with hexa\u00adnes. Several days later, X-ray quality black block-shaped crystals were collected.bCrystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2414314621005319/bh4061sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314621005319/bh4061Isup2.hklStructure factors: contains datablock(s) I. DOI: 2068473CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The study investigated clinical features of sellar and suprasellar tumors with optic nerve bending. Twenty-five patients with optic nerve bending in one eye who underwent tumor resection for sellar and suprasellar tumors were included. The other eye, without optic nerve bending, was the control. The pre- and postoperative best-corrected visual acuity (BCVA) and ganglion cell layer (GCL)\u2009+\u2009inner plexiform layer (IPL) thickness were studied retrospectively using optical coherence tomography. Preoperative BCVA in the eye with optic nerve bending was significantly poor and improved significantly after tumor resection. Eyes with optic nerve bending had significantly less GCL\u2009+\u2009IPL thickness on the temporal side than eyes without optic nerve bending. Preoperative GCL\u2009+\u2009IPL thickness of the entire macula was reduced in eyes with optic nerve bending and poor postoperative BCVA compared to those with good postoperative BCVA. There was no significant difference in GCL\u2009+\u2009IPL thickness of eyes with optic nerve bending before and after tumor resection. Optic nerve bending caused by sellar and suprasellar tumors resulted in visual impairment and decreased retinal ganglion cells. Eyes with optic nerve bending and severely reduced GCL\u2009+\u2009IPL thickness may have less BCVA improvement after tumor resection. Visual field impairment progresses gradually as the tumor grows, resulting in visual impairment3. However, even a relatively slight compression of the optic chiasm can cause visual impairment. We investigated the visual acuity impairment in the patients with sellar and suprasellar lesions using magnetic resonance (MR) T2\u00a0weighted imaging4. We also reported a new concept that the optic nerve sometimes bends at the entrance of the optic canal from the intracranial subarachnoid space. Further, a combination of preoperative optic nerve bending and optic chiasm compression is associated with visual impairment can measure ganglion cell layer (GCL)\u2009+\u2009inner plexiform layer (IPL) thickness in the macula with high reliability. Previous reports showed the usefulness of macular GCL\u2009+\u2009IPL parameters in discriminating early glaucoma from normal eyes5. Morphological changes in retinal ganglion cells might also be observed using OCT in patients with sellar and suprasellar tumors with optic nerve bending. Here, we investigated the OCT findings of patients with sellar and suprasellar tumors with optic nerve bending before and after tumor resection.Sellar and suprasellar tumors, including pituitary adenomas, show visual field defects, such as bilateral hemianopsia, due to the optic chiasm compression by the tumorTable All 25 patients with optic nerve bending were classified by age into groups of 30\u201340\u00a0s (n\u2009=\u20096), 50\u00a0s (n\u2009=\u20097), 60\u00a0s (n\u2009=\u20095), and 70\u00a0s (n\u2009=\u20097). Postoperative BCVAs (logMAR) were \u2212\u00a00.08\u2009\u00b1\u20090.00 (30\u201340\u00a0s),0.01\u2009\u00b1\u20090.17 (50\u00a0s), 0.08\u2009\u00b1\u20090.31 (60\u00a0s) and 0.13\u2009\u00b1\u20090.25 (70\u00a0s), respectively. Postoperative BCVA showed no significant differences among these 4 groups .The preoperative GCL\u2009+\u2009IPL thicknesses in bending and non-bending eyes were 65.6\u2009\u00b1\u200912.8\u00a0\u00b5m and 71.4\u2009\u00b1\u20099.7\u00a0\u00b5m in the superior sector, 64.8\u2009\u00b1\u200913.1\u00a0\u00b5m and 68.8\u2009\u00b1\u200911.1\u00a0\u00b5m in the superior nasal sector, 61.9\u2009\u00b1\u200912.4\u00a0\u00b5m and 64.8\u2009\u00b1\u200910.6\u00a0\u00b5m in the inferior nasal sector, 63.8\u2009\u00b1\u200912.9\u00a0\u00b5m and 70.0\u2009\u00b1\u20098.4\u00a0\u00b5m in the inferior sector, 72.9\u2009\u00b1\u200912.8\u00a0\u00b5m and 80.6\u2009\u00b1\u200911.5\u00a0\u00b5m in the inferior temporal sector, and 69.9\u2009\u00b1\u200911.8\u00a0\u00b5m and 75.8\u2009\u00b1\u20098.0\u00a0\u00b5m in the superior temporal sector, respectively. The postoperative GCL\u2009+\u2009IPL thicknesses in the bending and non-bending eyes were 65.2\u2009\u00b1\u200912.7\u00a0\u00b5m and 71.0\u2009\u00b1\u20099.1\u00a0\u00b5m in the superior sector, 63.6\u2009\u00b1\u200913.7\u00a0\u00b5m and 68.2\u2009\u00b1\u200910.7\u00a0\u00b5m in the superior nasal sector, 60.8\u2009\u00b1\u200912.6\u00a0\u00b5m and 64.6\u2009\u00b1\u200910.5\u00a0\u00b5m in the inferior nasal sector, 64.6\u2009\u00b1\u200911.4\u00a0\u00b5m and 68.4\u2009\u00b1\u20098.2\u00a0\u00b5m in the inferior sector, 72.3\u2009\u00b1\u200912.7\u00a0\u00b5m and 79.9\u2009\u00b1\u200912.3\u00a0\u00b5m in the inferior temporal sector, and 69.1\u2009\u00b1\u200912.4\u00a0\u00b5m and 75.6\u2009\u00b1\u20098.0\u00a0\u00b5m in the superior temporal sector, respectively of 0 or better as good visual outcome and 6 eyes with postoperative BCVA (logMAR) of less than 0 as poor visual outcome. The preoperative GCL\u2009+\u2009IPL thicknesses in the good visual outcome eyes and the poor visual outcome eyes were 69.9\u2009\u00b1\u20098.1\u00a0\u00b5m and 52.0\u2009\u00b1\u200915.3\u00a0\u00b5m in the superior sector, 68.7\u2009\u00b1\u200910.5\u00a0\u00b5m and 52.3\u2009\u00b1\u200912.8\u00a0\u00b5m in the superior nasal sector, 65.7\u2009\u00b1\u20099.6\u00a0\u00b5m and 49.8\u2009\u00b1\u200912.5\u00a0\u00b5m in the inferior nasal sector, 68.9\u2009\u00b1\u20096.6\u00a0\u00b5m and 47.8\u2009\u00b1\u200914.6\u00a0\u00b5m in the inferior sector, 76.9\u2009\u00b1\u20096.7\u00a0\u00b5m and 60.0\u2009\u00b1\u200917.9\u00a0\u00b5m in the inferior temporal sector, and 73.6\u2009\u00b1\u20096.7\u00a0\u00b5m and 58.2\u2009\u00b1\u200916.0\u00a0\u00b5m in the superior temporal sector, respectively analysis, of sellar and suprasellar tumors with sagittal bending of the optic nerve and compared them with those of non-bending optic nerve controls. Eyes with optic nerve bending due to sellar and suprasellar tumors had worse visual acuity and reduced GCL\u2009+\u2009IPL thickness in the temporal sectors, as measured by OCT, than eyes without optic nerve bending. In addition, eyes with optic nerve bending showed rapid improvement in visual acuity after tumor resection. Furthermore, in six eyes with poor visual outcome, the preoperative GCL\u2009+\u2009IPL thickness was significantly lesser than that in 19 eyes with good visual outcome.4. ONCBA basically affects ipsilateral vision4. Moreover, ipsilateral ONCBA is anatomically unrelated to contralateral visual dysfunction. However, when the ONCBA is large, the tumor is often large; therefore, the visual field defect due to chiasma compression may occur bilaterally. In addition, if the tumor is larger, the ONCBA on the contralateral side may be large. In this study, eyes with optic nerve bending had preoperative visual impairment, whereas eyes without optic nerve bending had good preoperative visual acuity , the preoperative OCT of the optic nerve bending eye showed only a mild decrease in GCL\u2009+\u2009IPL in the predominantly superior nasal sector, and the postoperative BCVA improved made evaluations, and any disagreements regarding conclusions were resolved by consensus. Optic nerve bending (large ONCBA) was defined as ONCBA\u2009\u2265\u200945\u00b0, and non-optic nerve bending (moderate ONCBA) was defined as ONCBA\u2009<\u200945\u00b0, as previously reported4. The exclusion criteria were as follows: (1) patients with a history of glaucoma or evident glaucomatous optic neuropathy; (2) high myopia (refractive error less than\u2009\u2212\u20096 diopters); (3) retinal diseases, including epiretinal membrane and macular edema; (4) severe cataract, and (5) unclear optic nerve on MR imaging.All experiments followed the tenets of the Declaration of Helsinki and were approved by the Institutional Review Board of the Gunma University Graduate School of Medicine (HS2021-097). Informed consent was obtained from all individual participants in the present study. We retrospectively studied 25 patients with visual impairment due to sellar and suprasellar tumors who underwent endoscopic transsphenoidal tumor resection at Gunma University Hospital from June 2015 to July 2021 and had optic nerve bending in only one eye. The other eye, without optic nerve bending, was used as the control. MR imaging of the sellar and suprasellar lesions in all 25 patients with a 1.5\u00a0T or 3\u00a0T MR imaging system was performed. The presence of optic nerve bending was determined by measuring the sagittal ONCBA on MR images before tumor resection, as previously reported5.All patients underwent ophthalmologic examinations, including best-corrected visual acuity (BCVA), intraocular pressure assessment, refraction, slit-lamp biomicroscopy, fundus examination, and GCL\u2009+\u2009IPL thickness measurement, using Cirrus high definition-OCT in both the optic nerve bending and non-bending eyes before and 1\u00a0month after tumor resection Fig.\u00a0. BCVA waData are presented as the mean\u2009\u00b1\u2009standard deviation. An unpaired t-test was conducted to compare BCVA and GCL\u2009+\u2009IPL thickness measurements between the bending and non-bending eyes. The paired t-test was conducted to compare changes in BCVA and GCL\u2009+\u2009IPL thickness before and 1\u00a0month after surgery. The correction between BCVA and ONCBA was examined using Spearman\u2019s correlation coefficient. Statistical significance was set at p\u2009<\u20090.05. Statistical analyses were performed using GraphPad Prism version 6 ."} +{"text": "In the crystal, ion pairs are linked by C\u2014H\u22efBr and N\u2014H\u22efBr hydrogen bonds and are connected into helical chains extending along the c-axis direction by weak, electrostatic S\u22efBr\u2212 inter\u00adactions. A Hirshfeld surface analysis was performed, which showed the dominant role of H\u22efH contacts (51.3%).In the title mol\u00adecular salt, C The dihedral angle between the mean planes of the thia\u00adzole and C2\u2013C7 rings is 13.1\u2005(1)\u00b0. A puckering analysis of the C1/C2/C7\u2013C10 ring yielded the parameters Q = 0.499\u2005(3)\u2005\u00c5, \u03b8 = 58.6\u2005(3)\u00b0 and \u03c6 = 225.6\u2005(3)\u00b0, indicating a half-chair conformation.As expected, the C11/C12/C13/N3/S1 thia\u00adzole ring in \u2212 contacts of this length or shorter are present in the Cambridge Structural Database, two examples being reported by Auffinger et al. .In the crystal, the S1\u22efBr1 distance of 3.5017\u2005(7)\u2005\u00c5 is some 0.15\u2005\u00c5 less than the sum of the van der Waals radii and likely represents an electrostatic inter\u00adaction between the two atoms since S1 is near to the cationic charge. Over 200 structures having S\u22efBr al. 2004 and Thom al. 2004. This ins Table\u00a01 form helon Fig.\u00a02. It may ts Fig.\u00a03, in agreet al., 2016A yielded 30 hits of which 11 were considered similar to I. Among these, (Z)-1--2-[1-(4-hy\u00addroxy\u00adphen\u00adyl)ethyl\u00adidene]hydra\u00adzinium bromide unknown solvate -2-[(2-nitro\u00adphen\u00adyl)methyl\u00adidene]-1-hydrazinium bromide -yl\u00adidene)hydrazinyl\u00adidene]eth\u00adyl}pyridinium bromide monohydrate . Fig.\u00a04a presents the Hirshfeld surface plotted over dnorm with a second cation closest to the bromide ion also present, clearly showing the N\u2014H\u22efBr and C\u2014H\u22efBr inter\u00adactions as well as the S1\u22efBr1 short contact (dashed lines). The surface plotted over shape and curvature indices do not show much flat surface or evidence for \u03c0-stacking inter\u00adactions, in agreement with the results given in Section 3. Fig.\u00a05a) and resolved into all H\u22efH contacts , H\u22efC/C\u22efH contacts , Br\u22efH/H\u22efBr contacts and S\u22efH/H\u22efS contacts . The N\u22efH/H\u22efN contacts contribute only 1.3%.The Hirshfeld surface for pe Fig.\u00a04b and cues Fig.\u00a04c do notet al., 2013I suitable for X-ray diffraction were obtained by recrystallization of the crude product from ethanol solution.The title compound was prepared according to our previously reported method global, I. DOI: 10.1107/S2056989021002863/hb7963Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989021002863/hb7963sup3.pdfGeometrical data for title compound and related phases. DOI: Click here for additional data file.10.1107/S2056989021002863/hb7963Isup5.cmlSupporting information file. DOI: 2071135CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of 1 consists of two crystallographically independent TPRS-{NdIIIO9} units that form an edge-sharing dinuclear cluster inter\u00adconnected to neighbouring dimers by the \u03bc3-SO42\u2013 anions, yielding a cationic two-dimensional {Nd2(H2O)6(SO4)2}nn+2 sheet. Adjacent cationic layers are then linked via the \u03bc4-glutarato2\u2013 ligands into a three-dimensional coordination network. Strong O\u2014H\u22efO hydrogen bonds are the predominant inter\u00adaction in the crystal structure.A three-dimensional coordination polymer, poly[hexa\u00adaqua\u00ad(\u03bc These properties are cooperatively provided by both the inorganic building units and the organic counterparts (Furukawa 2(H2O)6(glutarato)(SO4)2]n (1). The crystal structure reveals that the glutarates act as bridging ligands binding the cationic {Nd2(H2O)6(SO4)2}nn+2 sheets into a three-dimensional network.Herein, we report a microwave synthesis of a new three-dimensional coordination polymer, n crystallizes in the monoclinic P21/c space group. There are two crystallographically independent NdIII cations (Nd1 and Nd2), two sulfate anions, and six coordinated water mol\u00adecules in the asymmetric unit, as illustrated in Fig.\u00a01The coordination network III cations are nine-coordinated to O atoms from one bridging glutarate2\u2212, two chelating glutarate2\u2212, two chelating sulfate anions and three coordinated H2O, adopting a distorted tricapped trigonal\u2013prismatic geometry, TPRS-{NdIIIO9} , forming an edge-sharing dinuclear unit with its symmetry-related NdIIIO9 polyhedron. The NdIII\u2014O bond distances are in the range of 2.383\u2005(2)\u20132.785\u2005(2)\u2005\u00c5, which are reasonable and comparable to those reported for other NdIII coordination polymers such as [Nd(H2O)4(glutarato)]Cl 4(glutarato)]Cl\u00b72H2O 2(glutarato)]\u00b72H2O 4]Cl 4]Cl\u00b72H2O x(glutarato)}nn+ subunits compensated by uncoordinated chloride anions, each of the tetra\u00adhedral SO42\u2013 ligands in 1 links three adjacent NdIII atoms, forming a neutral two-dimensional network of [Nd2(H2O)6(glutarato)(SO4)2]n. The S\u2014O bond distances are in the range 1.449\u2005(3)\u20131.485\u2005(2)\u2005\u00c5, with O\u2014S\u2014O angles ranging from 107.78\u2005(16) to 111.67\u2005(15)\u00b0. The flexible glutarate linker exhibits a .Both Nd1 can be described as a three-dimensional non-porous framework, which is constructed from edge-sharing TPRS-{NdIIIO9} polyhedra linked through sulfate anions, acting as tritopic inorganic linkers, into a cationic [Nd2(H2O)6(SO4)2]nn+2 sheets parallel to the (011) layers, as illustrated in Fig.\u00a03a. It is noteworthy that these sheets also contain large inorganic [Nd(SO4)]4 rings further stabilized by O\u2014H\u22efO hydrogen bonds between the water mol\u00adecules and sulfate anions (Table\u00a01b). This three-dimensional arrangement also features O\u2014H\u22efO hydrogen bonds between two water mol\u00adecules or between a water mol\u00adecule and oxygen atoms of the glutarate ligands . In total, all but one hydrogen atom from the six crystallographically independent water mol\u00adecules are involved in hydrogen bonding tetra\u00adaqua\u00adneo\u00addym\u00adium chloride] tetra\u00adaqua\u00addineodymium chloride dihydrate] di\u00adaqua\u00addi\u00adneo\u00addym\u00adium(III) tetra\u00adhydrate] diaqua\u00addi\u00adneodymium(III) dihydrate] 3\u00b78H2O , glutaric acid , and 4,4\u2032-bi\u00adpyridine in 40.0\u2005mL of deionized water under ambient conditions. The solution was transferred into an open glass reactor and then irradiated by microwaves (800\u2005W) for 10 minutes. The solution was let to cool to ambient temperature. Pale-purple block-shaped crystals crystallized from the solution within a few minutes. FT\u2013IR of 1: stretch\u03bd(O\u2014H) 3364, stretch\u03bd(C\u2014H) 2990, as\u03bd(COO\u2212) 1531, s\u03bd(COO\u2212) 1430, \u03b4(O\u2014H) 1355, s\u03bd(S\u2014O) 1101, s\u03bd(S\u2014O) 1077, s\u03bd(SO42\u2013) 596.Complex Uiso(H) = 1.2Ueq(C). The H atoms from the water mol\u00adecules were located in the residual electron-density map, and where necessary, refined with distance and angle restraints or riding on the parent oxygen atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022000159/jq2009sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022000159/jq2009Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022000159/jq2009Isup3.molSupporting information file. DOI: 2107848CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Pistacia vera is one of the most important cash crops in Iran that is scattered in arid and semi-arid regions. During a survey of plant-parasitic nematodes of pistachio in Ardakan city in Yazd Province, a species of root-lesion nematode was isolated and identified by morphological, morphometrical, and molecular methods as Pratylenchus oleae Palomares-Rius et al., 2014. This species was isolated from several pistachio trees rhizosphere regarding to molecular analysis, D2\u2013D3 expansion segments of 28S rRNA was amplified by PCR and sequenced. The sequence was deposited in GenBank (Accession No. MW338666). Along with the related phylogenetic analysis, placed this species in a monophyletic clade with other Pratylenchus oleae isolates, based on Bayesian inference (BI) phylogeny. A PCR reaction with the P. oleae specific primer set produced a 547\u00a0bp fragment. This is the first report of P. oleae infecting Pistachio tree in the world.Pistachio, Pistacia. P. vera is one of the most important commercial cultivars among the 11 identified species. More than 498,000 hectares of Iranian lands are cultivated with pistachio trees and its product with nearly 173000 tones has a high domestic and foreign acceptance and crushed with a microhomogeniser Vibro Mixer . Two microliters proteinase K (600\u00a0\u03bcg/ml) were added and the tubes were frozen at \u201280\u00b0C for at least 10\u2009\u2009min and then incubated at 65\u00b0C (1 hr) and 95\u00b0C (10\u2009\u2009min) consecutively. After incubation, the tubes were centrifuged for 2\u2009\u2009min at 14,000\u2009\u2009rpm and kept at \u201220\u00b0C until use. The PCR was carried out in a 30\u2009\u2009\u03bcl reaction comprising of 2\u2009\u2009\u00b5L DNA template, 1\u2009\u2009\u03bcl forward and reverse primers, 15\u2009\u2009\u03bcl Taq DNA Polymerase 2\u00a0\u00d7\u00a0Master Mix (Ampliqon), and 11\u2009\u2009\u03bcl distilled water. The forward primer D2A (5\u2032-ACAAGTACCGTGA GGGAAAGTTG-3\u2032) and the reverse primer D3B (5\u2032-TCGGAAGGAACCAGCTACTA-3\u2032) (P. oleae-specific primer pairs Poleae_fw1_4_36 (5\u2032-GACAGATTAGAATGGAATCTGTTCG-3\u2032) and Poleae_rv1_ _525_551 (5\u2032ATCGCTTTTGGATTCAATAATATA-3\u2032) as described by P. oleae sequence was aligned with related sequences from GenBank through National Center for Biotechnology Information (NCBI) BLASTn homology search, using ClustalW implemented by MEGA version 10.0. The GTR\u00a0+\u00a0I\u00a0+\u00a0G model was selected as the best by jModeltest v.2.1.10. Bayesian tree generated using the Bayesian inference method as implemented in the program MrBayes v.3.2.6 were use v.3.2.6 . Markov datasets .P. oleae, found in roots and around soil from Pistachio tree in Iran are presented in P. oleae as well. P. oleae is characterized by morphological features in females: Vermiform body, lip region with three annuli, slightly offset. Stylet well developed with distinct rounded knobs slightly directed anteriorly. Lateral field with four incisures, metacorpus oval to rounded, isthmus rather short, surrounded by nerve ring, pharyngeal glands well developed, with rather long ventral overlap. Excretory pore either slightly anterior pharyngo-intestinal junction or opposite. Genital branch with one row of oocyte, spermathecal rounded, vagina a straight tube. Vulva transverse slit. Post-vulval uterine sac short, not differentiated. Tail sub-cylindrical, terminus rounded to conical, smooth, male unknown : L\u2009\u2009=\u2009\u2009522.5 \u00b1\u00a034.9 (463.0\u2013565.0) \u03bcm; a\u2009\u2009=\u2009\u200928.3\u00a0\u00b1\u00a01.6 (25.7\u201331.0)\u2009\u2009\u03bcm; b\u2009\u2009=\u2009\u20095.9\u00a0\u00b1\u00a00.4 (5.2\u20136.7)\u2009\u2009\u03bcm; c\u2009\u2009=\u2009\u200920.2\u00a0\u00b1\u00a00.8 (19.1\u201321.0) \u03bcm; Stylet length\u2009\u2009=\u2009\u200917.3\u00a0\u00b1\u00a00.7 (16.0\u201318.0) \u03bcm; Median bulb 55.6\u00a0\u00b1\u00a02.3 (51.0\u201359.0) \u03bcm; Anterior to Excretory Pore 88.9\u00a0\u00b1\u00a02.7 (85.0\u201393.0) \u03bcm; Anterior end to pharyngeal junction 89.0\u00a0\u00b1\u00a04.6 (78.0\u201394.0) \u03bcm; Pharynx Length 123.5\u00a0\u00b1\u00a03.4 (118.0\u2013129.0) \u03bcm; Max. Body diam. 18.5 \u00b1\u00a01.0 (17.0\u201320.0) \u03bcm; Body diam. at anus 12.6\u00a0\u00b1\u00a00.5 (12.0\u201313.0)\u2009\u2009\u03bcm and tail length\u2009\u2009=\u2009\u200925.9\u00a0\u00b1\u00a01.7\u2009\u2009(22.0\u201327.0)\u2009\u2009\u03bcm \u2009\u2009\u03bcm .P. oleae belong to Spain and Tunisia . Pratylenchus capsici, 9 isolates are most similar to P. oleae, but differs in presence of males, a functional spermatheca, a larger body and shorter stylet with P. oleae Spain isolate (KJ510861) and 99% (2\u20137 nucleotide differences) similar to P. oleae Spain isolates and P. oleae Tunisia isolates , while P. dunensis and P. penetrans are 89% and 87% similar (61 and 82 nucleotide differences with 6 and 13 Gaps). The gap variations and nucleotide differences between 9 isolates of P. capsici and P. oleae (MW338666) are between 3\u20139 and 30\u201337 nucleotides respectively and one location of Tunisia (Ouled Chamekh) , But somP. oleae deposited in GenBank database. The test population had no gap (base pair), which was different to those of P. oleae from Spain (KJ510855 and KJ510861) with 100% similarity, one gap with Spain (KJ510856 and KJ510857) and Tunisia (KJ510858) with 99% similarity respectively and two gaps with Tunisia (KJ510859 and KJ510860) with 99% similarity. In summary, Iranian P. oleae was isolated from pistachio trees in Ardakan, Yazd province. It is reported for the first time from Iran and pistachio tree in the world. This nematode can be considered as a risk to the economy in pistachio orchards.The sequence of Iranian isolate showed high nucleotide similarity with D2-D3 region of"} +{"text": "The Mg atoms in the title compound are located at an inter\u00adstitial site of the Dy2.1B37Si9-type structure with an occupancy of 0.5. The (001) layers of B12 icosa\u00adhedra stack along the c-axis direction with shifting in the direction. A three-dimensional framework structure of the layers is formed via B\u2014Si bonds and {Si8} units of [Si]3\u2014Si\u2014Si\u2014[Si]3.Single crystals of a novel sodium\u2013magnesium boride silicide, Na The structure is composed of B12 icosa\u00adhedral clusters: the B\u2014B distances of the 30 distinct bonds in the cluster are in the range of 1.791\u2005(3)\u20131.843\u2005(5)\u2005\u00c5 and the average distance is 1.811\u2005\u00c5 \u2005\u00c5] on the (001) plane and form layers that stack along the c axis with a sequence of ABCABC by shifts of \u2005\u00c5 for Si3\u2014Si3 and 2.3951\u2005(9)\u2005\u00c5 for Si2\u2014Si3 are comparable with the bond length in crystalline silicon (2.35\u2005\u00c5). The bond angles of Si2\u2014Si3\u2014Si2 and Si2\u2014Si3\u2014Si3 are 113.86\u2005(3)\u00b0 and 104.61\u2005(4)\u00b0, respectively, which are distorted from the regular tetra\u00adhedral bond angle of 109.47\u00b0. The Si2\u2014B1 distance is 2.043\u2005(2)\u2005\u00c5, which is close to the Si\u2014B distances (1.973\u20132.027\u2005\u00c5) found in \u03b2-silicon boride, SiB3 \u2005\u00c5] and Si1/B5\u2014Si1/B5 pairs that bind to the B atoms at B3 connect the B12 layers of Na3MgB37Si9 .The framework structure of Bi9 Fig.\u00a01. Because8} unit between the B12 cluster layers. The Na1\u2014Si2 distance is 2.8620\u2005(4)\u2005\u00c5 and the Na1\u2014B1 and Na1\u2014B2 distances are 2.811\u2005(2) and 2.793\u2005(2)\u2005\u00c5, respectively. These distances are almost the same as the Na\u2014Si distance of Na4Si4 [2.878\u2005(3)\u2005\u00c5; Morito et al., 201515 \u2005\u00c5 and 2.333\u2005(3)\u2005\u00c5, respectively, which are close to the Mg\u2014Si (2.436\u2005\u00c5) and Mg\u2014B distances (2.353\u2005\u00c5) in MgB12Si2 provide six and 6.3 electrons, respectively, and approximately six electrons are supplied from RE in REx1\u2013B12Si\u03b43.3\u2013 and REx1\u2013B36Si9C . The lattice constants and unit-cell volume of Mg3B36Si9C are a = 10.0793\u2005\u00c5, c = 16.372\u2005\u00c5, and V = 1440.4\u2005\u00c53 are a = 10.046\u201310.095\u2005\u00c5, c = 16.298\u201316.467\u2005\u00c5, and V = 1429\u20131454\u2005\u00c53 are a = 10.000\u201310.096\u2005\u00c5, c = 16.225\u201316.454\u2005\u00c5, and V = 1405\u20131452\u2005\u00c53 . Then, 10\u2005mg of B2O3 powder were added to the crucible, which was stacked on another BN crucible containing 30\u2005mg of Mg powder , and these crucibles were encapsulated in a stainless steel container with Ar gas. The container was heated at 1373\u2005K for 24\u2005h using an electric furnace. After cooling, the crucible was taken out from the reaction container, and any Na and NaSi remaining in the crucible were reacted and removed with 2-propanol and ethanol. Then, the sample was washed with pure water to remove water-soluble compounds such as sodium borate and alkoxide produced by the reaction of Na and alcohol to leave black plates of the title compound. An electron probe microanalyzer was used to analyze the composition of the obtained single crystal as Na 5.49\u2005(8), Mg 2.37\u2005(7), B 74.8\u2005(7), Si 17.3\u2005(4) atom %, which is nearly matched by Na3MgB37Si9 . Other elements such as O were not found.Na metal pieces , crystalline B powder and Si powder were weighed in a BN crucible , with a molar ratio of Na:B:Si = 5:4:3 in a high-purity Ar-filled glove box twinning, which reduced the R-value from 0.0651 to 0.0380.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022000494/hb8005sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022000494/hb8005Isup2.hklStructure factors: contains datablock(s) I. DOI: 2141726CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(6) ring motif. Inter\u00admolecular N\u2014H\u22efN and C\u2014H\u22efN hydrogen bonds, as well as N\u2014H\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions create a three-dimensional network in the crystal.The mol\u00adecular conformation of the title compound is stabilized by an intra\u00admolecular O\u2014H\u22efO hydrogen bond, forming an 20H15Cl2N3O2, is stabilized by an intra\u00admolecular O\u2014H\u22efO hydrogen bond, forming an S(6) ring motif. The central pyridine ring is almost planar [maximum deviation = 0.074\u2005(3)\u2005\u00c5]. It subtends dihedral angles of 86.10\u2005(15) and 87.17\u2005(14)\u00b0, respectively, with the phenyl and di\u00adchloro\u00adphenyl rings, which are at an angle of 21.28\u2005(15)\u00b0 to each other. The =C(\u2014OH)CH3 group is coplanar. In the crystal, mol\u00adecules are linked by inter\u00admolecular N\u2014H\u22efN and C\u2014H\u22efN hydrogen bonds, and N\u2014H\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional network. The most important contributions to the crystal packing are from H\u22efH (33.1%), C\u22efH/H\u22efC (22.5%), Cl\u22efH/H\u22efCl (14.1%), O\u22efH/H\u22efO (11.9%) and N\u22efH/H\u22efN (9.7%) inter\u00adactions.The mol\u00adecular conformation of the title compound, C A strong intra\u00admolecular O2\u2014H2\u22efO1 hydrogen bond ring motif create a three-dimensional network in the crystal in the range of \u22120.4290 to 1.5192 a.u. The red patches that appear around N2 are caused by the inter\u00admolecular N3\u2014H3A\u22efN2 and C16\u2014H16\u22efN2 inter\u00adactions, which are important in the packing of the title mol\u00adecule. Bright red dots near N2 and amine hydrogen atoms H3A and H3B highlight their functions as hydrogen-bonding acceptors and donors, respectively; these also appear as blue and red areas on the Hirshfeld surface mapped over electrostatic potential , C\u22efH/H\u22efC , Cl\u22efH/H\u22efCl , O\u22efH/H\u22efO and N\u22efH/H\u22efN . Other Cl\u22efC/C\u22efCl, C\u22efC, Cl\u22efO/O\u22efCl, Cl\u22efN/N\u22efCl, N\u22efC/C\u22efN, O\u22efN/N\u22efO, Cl\u22efCl, O\u22efC/C\u22efO and N\u22efN contacts contribute less than 2.1% to Hirshfeld surface mapping and have little directional influence on mol\u00adecular packing and SETWUK (space group: P21/n), adopt nearly planar structures. The crystal structure of SETWOE is stabil\u00adized by inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. The C\u2014H\u22efO hydrogen bonds generate rings with 22R(14) and 22R(20) motifs. The crystal structure of SETWUK is stabilized by inter\u00admolecular C\u2014H\u22efF and C\u2014H\u22ef\u03c0 inter\u00adactions. The C\u2014H\u22efF bond generates a linear chain with a C(14) motif. In addition, in SETWOE and SETWUK, intra\u00admolecular O\u2014H\u22efO inter\u00adactions are found, which generate an S(6) graph-set motif. No significant ar\u00adyl\u2013aryl or \u03c0\u2013\u03c0 inter\u00adactions exist in these structures. All this bears some resemblance to the title compound.The polysubstituted pyridines, SETWOE (space group: PP22R(8) ring motif. In the hydrated salt JEBREQ, the presence of the water mol\u00adecule prevents the formation of the familiar i.e. R32(8)] is formed involving the sulfonate group, the pyrimidinium cation and the water mol\u00adecule. Both salts form a supra\u00admolecular homosynthon . Their isotropic displacement parameters were refined using a riding model with Uiso(H) set to either 1.2Ueq(N) for the NH2 group or 1.5Ueq(O) for the OH group. The C-bound H atoms were positioned geometrically (C\u2014H = 0.95\u20131.00\u2005\u00c5) and allowed to ride on their parent atoms, with Uiso(H) = 1.5Ueq(C) for the methyl group and Uiso(H) = 1.2Ueq(C) for aromatic and methine H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989021007994/yz2010sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021007994/yz2010Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021007994/yz2010Isup3.cmlSupporting information file. DOI: 2101203CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "R/S,E)-2-(4-hy\u00addroxy\u00adphen\u00adyl)-4-(2-phenyl\u00adhydrazin-1-yl\u00adidene)chromane-5,7-diol ethanol monosolvate, has been determined. The packing is assisted by C\u2014H\u22efO, O\u2014H\u22efO, O\u2014H\u22efN and O\u2014H\u22efC(\u03c0) type hydrogen bonds, and the pyran ring has an envelope pucker.The structure of racemic -2-(4-hy\u00addroxy\u00adphen\u00adyl)-4-(2-phenyl\u00adhydrazin-1-yl\u00adidene)chromane-5,7-diol ethanol monosolvate, C21H18N2O4\u00b7C2H6O, in a centrosymmetric lattice is reported. The two racemates occupy the same position in the asymmetric unit \u2013 a disordered mixed enanti\u00adomeric structure. Hydrogen bonds of the type O\u2014H\u22efC(\u03c0) in addition to typical C\u2014H\u22efO, O\u2014H\u22efO and O\u2014H\u22efN are identified. A positional disorder is seen in the solvent mol\u00adecule (ethanol) as well. The phenyl\u00adhydrazone group is nearly coplanar with the chromane ring system [dihedral angle = 15.5\u2005(1)\u00b0], while the the 4-hy\u00addroxy\u00adphenyl ring is perpendicular [dihedral angle = 87.2\u2005(1)\u00b0] to the chromane. The pyran ring has an envelope pucker .The crystal structure of racemic ( Enanti\u00adomeric structures in centrosymmetric lattices have been discussed by Flack (2003Q = 0.363\u2005(3)\u2005\u00c5, \u03b8 = 57.6\u2005(3)\u00b0; and for the enanti\u00adomer: Q = 0.364\u2005(3)\u2005\u00c5, \u03b8 = 127.4\u2005(4)\u00b0]. An intra\u00admolecular O\u2014H\u22efN hydrogen bond exists between one of the hy\u00addroxy groups on the chromane ring and the nitro\u00adgen of the hydrazone group mol\u00adecule in 1:1 ratio, yielded a disordered mixed enanti\u00adomeric crystal in a centrosymmetric lattice type hydrogen-bond inter\u00adactions between the solvent ethanol and phenyl ring are observed Table\u00a01. The phe al. 1993. The strce Fig.\u00a02.et al., 2016Chemical context section. The most similar structures for which crystal data have been reported include acyl hydrazone derivatives of 2-phenyl\u00adchroman-4-one and hesperetin. In particular, crystal structures for 2\u2032-[2-(4-fluoro\u00adphen\u00adyl)chroman-4-yl\u00adidene]isonicotino\u00adhydra\u00adzide -amino}\u00adbenz\u00ada\u00admide was performed in the Cambridge Structural Database , 0.97\u2005\u00c5 (CH2), 0.86\u2005\u00c5 (NH) or 0.82\u2005\u00c5 (OH). Isotropic displacement parameters for these atoms were set to 1.2 or 1.5 times Ueq of the parent atom. Idealized Me of the ethanol mol\u00adecule were refined as rotating group(s): C22A and C22B (H22A through F) and its idealized tetra\u00adhedral OH refined as a rotating group: O5A and O5B .Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022002079/jy2016sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022002079/jy2016Isup3.molSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022002079/jy2016Isup3.cmlSupporting information file. DOI: 2153764CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Z-shaped conformation with the carboxyl group nearly coplanar with the di\u00adhydro\u00adquinoline unit. In the crystal, two sets of C\u2014H\u22efO hydrogen bonds form chains along the b-axis direction, which are connected into corrugated layers parallel to (103) by additional C\u2014H\u22efO hydrogen bonds. The layers are connected by C\u2014H\u22ef\u03c0(ring) inter\u00adactions.The mol\u00adecule adopts a 20H15NO3, adopts a Z-shaped conformation with the carboxyl group nearly coplanar with the di\u00adhydro\u00adquinoline unit. In the crystal, corrugated layers are formed by C\u2014H\u22efO hydrogen bonds and are stacked by C\u2014H\u22ef\u03c0(ring) inter\u00adactions. Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from H\u22efH (43.3%), H\u22efC/C\u22efH (26.6%) and H\u22efO/O\u22efH (16.3%) inter\u00adactions. The optimized structure calculated using density functional theory at the B3LYP/ 6\u2013311\u2005G level is compared with the experimentally determined structure in the solid state. The calculated HOMO\u2013LUMO energy gap is 4.0319\u2005eV.The title mol\u00adecule, C This moiety is planar to within 0.0340\u2005(6)\u2005\u00c5 (r.m.s. deviation = 0.0164) with N1 and C9 being, respectively, 0.0340\u2005(6) and \u22120.0279\u2005(7)\u2005\u00c5 from the mean plane, resulting in a slight twist at this location. The carboxyl group is nearly coplanar with the di\u00adhydro\u00adquinoline as seen from the 1.04\u2005(5)\u00b0 dihedral angle between the plane defined by C7/C13/O2/O3 and that of the di\u00adhydro\u00adquinoline (C1\u2013C9/N1/O1). This is likely due, in part, to the intra\u00admolecular C5\u2014H5\u22efO2 inter\u00adaction Table\u00a01. The prob-axis direction, which are connected by inversion-related pairs of C4\u2014H4\u22efO2 hydrogen bonds . The positive electrostatic potential (blue region) over the surface indicates hydrogen-donor potential, whereas the hydrogen-bond acceptors are represented by negative electrostatic potential (red region).The nd Fig.\u00a04b. The pet al., 2007a, while those delineated into H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH, C\u22efC, O\u22efC/C\u22efO, H\u22efN/N\u22efH, N\u22efC/C\u22efN and N\u22efO/O\u22efN contacts are illustrated in Fig.\u00a05b\u2013i, respectively, together with their relative contributions to the Hirshfeld surface (HS). The most important inter\u00adaction is H\u22efH, contributing 43.3% to the overall crystal packing, which is reflected in Fig.\u00a05b as widely scattered points of high density due to the large hydrogen content of the mol\u00adecule, with its tip at de = di = 1.19\u2005\u00c5. In the presence of C\u2014H inter\u00adactions, the pair of characteristic wings in the fingerprint plot delineated into H\u22efC/C\u22efH contacts has tips at de + di = 3.07\u2005\u00c5. The pair of scattered points of spikes in the fingerprint plot delineated into H\u22efO/O\u22efH contacts have tips at de + di = 2.08\u2005\u00c5. The C\u22efC contacts have tips at de + di = 3.34\u2005\u00c5. The O\u22efC/C\u22efO contacts, Fig.\u00a05f, contribute 1.5% to the HS and appear as a pair of scattered points of spikes with tips at de + di = 3.55\u2005\u00c5. The H\u22efN/N\u22efH contacts have tips at de + di = 3.28\u2005\u00c5. Finally, the C\u22efN/N\u22efC and O\u22efN/N\u22ef O contacts, Fig.\u00a05h\u2013i, contribute only 0.5% and 0.1% respectively to the HS and have a low-density distribution of points.The overall two-dimensional fingerprint plot , hardness (\u03b7), ionization potential (I), dipole moment (\u03bc), electron affinity (A), electrophilicity (\u03c9) and softness (\u03c3) are collated in Table\u00a03\u0394E = ELUMO\u00a0\u2212\u00a0EHOMO) of the mol\u00adecule is 4.0319\u2005eV, and the frontier mol\u00adecular orbital energies, EHOMO and ELUMO, are \u22126.3166 and \u22122.2847\u2005eV, respectively.The structure in the gas phase of the title compound was optimized by means of density functional theory. The density functional theory calculation was performed by the hybrid B3LYP method and the 6\u2013311\u2005G basis-set, which is based on Becke\u2019s model , benzyl chloride and tetra n-butyl\u00adammonium bromide as a catalyst in DMF (30\u2005mL) was stirred at room temperature for 48\u2005h. After removal of the salts by filtration, the solvent was evaporated under reduced pressure and the residue obtained was dissolved in di\u00adchloro\u00admethane. The organic phase was dried over Na2SO4 and concentrated under vacuum. The crude product obtained was purified by chromatography on a column of silica gel (eluent: hexa\u00adne/ ethyl acetate: 9/1). 1H NMR \u03b4 ppm: 3.08 ; 4.37 ; 5.12 ; 7.08\u20138.74 ; 13C NMR \u03b4 ppm: 34.3 (CH3\u2014N); 66.2 (CH2\u2014O); 72.1 (\u2013C\u2261); 73.2 (CH\u2261); 115.6-148.7 (CHarom and Cquat arom); 162. 5 (C=Oquinol); 168.2 (C=Ocarbox\u00adyl). MS (ESI): m/z = 318 (M + H)+.A mixture of 2-oxo-1-(prop-2-yn-1-yl)-1,2-di\u00adhydro\u00adquinoline-4-carb\u00adoxy\u00adlic acid , KCrystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989021007416/tx2040sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989021007416/tx2040Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021007416/tx2040Isup3.cmlSupporting information file. DOI: 2097267CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-021-02868-x, published online 10 December 2021Correction to: The original version of this Article contained an error in the legend of Figure\u00a01.a) Garadagh mud volcano in onshore Azerbaijan (40.24\u00b0N and 49.51\u00b0E).\u201d\u201c(now reads:(a) Otmanbozdag mud volcano (within the Garadagh structure) in onshore Azerbaijan (40.24\u00b0N and 49.51\u00b0E).\u201d\u201cThe original Article has been corrected."} +{"text": "This compound crystallized in the space group P\u03b75-coordinated Cp\u2032 rings and a \u03ba3-coordinated (BH4)\u2212 ligand.New syntheses have been developed for the synthesis of (borohydrido-\u03ba For example, \u03ba1, \u03ba2, and \u03ba3 U\u2013(BH4) binding has previously been reported 4 (Cp\u2032 = C5H4SiMe3), made in 1992 \u2212 within the tris-cyclo\u00adpenta\u00addienyl uranium platform using single-crystal X-ray diffraction. Toward this end, we developed new synthetic routes to the Cp\u20323U(BH4) compound.Actinide borohydrides have been of inter\u00adest since the 1940s, owing to their potential volatility and applied use in vapor deposition technologies for the production of thin films compound was originally synthesized by reacting Cp\u20323UH with H3B-PPh3 from Cp\u20323UH with H3B-PPh3 in hot THF solvent. We also observed Cp\u20323U(BH4) could be prepared in high yield (96%) by reacting Cp\u20323UI with NaBH4 in the presence of 15-crown-5. When this reaction was carried out in toluene at room temperature, the I\u2212 ligand was substituted by the (BH4)\u2212 anion. Another method we developed for synthesizing Cp\u20323U(BH4) involved reacting U(BH4)4 with KCp\u2032 (3 equiv.) in diethyl ether. This reaction, where (BH4)\u2212 was substituted by (Cp\u2032)\u2212, also proceeded in high yield (89%). X-ray quality crystals of Cp\u20323U(BH4) formed at 253\u2005K overnight from diethyl ether solutions.The Cp\u20323U(BH4) from Cp\u20323UI over U(BH4)4 because the U(BH4)4 starting material was more challenging to isolate in a chem\u00adic\u00adally pure form. Another inter\u00adesting comparison between the two synthetic methods involved the substitution chemistry. The (Cp\u2032)\u2212 anion displaced (BH4)\u2212 from U(BH4)4 and (BH4)\u2212 displaced I\u2212 in Cp\u20323UI. Hence, we qualitatively concluded that the stability of the U\u2014X bond for mol\u00adecular compounds dissolved in organic solvents was largest for (Cp\u2032)\u2212, inter\u00admediate for (BH4)\u2212, and lowest for I\u2212. The generality of this conclusion is limited, and we acknowledge the solubility of the other reaction products (such as NaI) might significantly influence the substitution chemistry on uranium.Of our two synthetic routes, we preferred making Cp\u20323U(BH4) were refined in the triclinic Pet al., 20104)3(DME)]2(\u03bc-O) , 2.574\u2005(6), 2.584\u2005(6), and 2.635\u2005(7)\u2005\u00c5 3U(BH4) was reported to be 2.48\u2005\u00c5 3U(BH4) in the gas phase and in solution predicted U\u2014B distances of 2.533 and 2.557\u2005\u00c5 2U(BH4)2 structures showed similar U\u2014B distances of 2.56\u2005(1)\u2005\u00c5 for [C5H3(SiMe3)2]2U(BH4)2 2U(BH4)2 2U(BH4)2 range from 2.458\u20132.500\u2005\u00c5 and average 2.48\u2005(2)\u2005\u00c5 (uncertainty reported as the standard deviation from the mean at 1\u03c3). These uranium\u2013(Cp\u2032 centroid) distances compare well with the 2.473\u2005\u00c5 analogous metric in Cp\u20323UCl 3] is more acute than the 117.0\u00b0 angle in Cp\u20323UCl and other Cp\u20323UX structures, where the average (Cp\u2032 centroid)\u2014U\u2014(Cp\u2032 centroid) angles were reported as 117\u2005(1)\u00b0 in Cp\u20323UI, 112\u2005(2)\u00b0 in Cp\u20323U(\u03b71-CH=CH2), and 118.7\u2005(4)\u00b0 in Cp\u20323U[Si(SiMe3)3]. The more acute (Cp\u2032 centroid)\u2014U\u2014(Cp\u2032 centroid) angles are complemented by a more obtuse average (Cp\u2032 centroid)\u2014U\u2014B angle of 104.4\u2005(4)\u00b0 in Cp\u20323U(BH4), likely due to the close proximity of the (BH4)1\u2212 ligand compared with (Cp\u2032 centroid)\u2014U\u2014X angles of 100.0\u00b0 in Cp\u20323UCl, 100\u2005(2)\u00b0 in Cp\u20323UI, 98\u2005(3)\u00b0 in Cp\u20323U(\u03b71-CH=CH2), and 96.7\u2005(9)\u00b0 in Cp\u20323U[Si(SiMe3)3], see Table\u00a01The uranium\u2013(Cp\u2032 centroid) distances in Cp\u20323U(BH4) structure is that all three of the tri\u00admethyl\u00adsilyl groups are oriented in a single direction towards the (BH4)\u2212 unit. This orientation has not been observed in other Cp\u20323U(anion) and Cp\u20323U(\u03bc-dianion)UCp\u20323 structures, which are shown in Figs. 23UCl structure 3] complex 3]1\u2212 unit. Since Cp\u20323U(BH4) has the smallest mono-anion of the Cp\u20323U(anion) complexes and the correspondingly smallest (Cp\u2032 centroid)\u2014U\u2014(Cp\u2032 centroid), and the largest (Cp\u2032 centroid)\u2014U\u2014X angles, the orientation of the silyl groups could occur due to steric factors. However, it is also possible that some dispersion forces between the (BH4)\u2212 and the tri\u00admethyl\u00adsilyl groups could contribute to the orientation piano-stool complexes: (C5H5)U(BH4)3 U(BH4)(SPSMe) Ph2)-1-Me, a \u03bb4-phosphinine with two lateral phosphino\u00adsulfide groups, and the tetra\u00admethyl\u00adphosphol (PC4Me4) compound (PC4Me4)(C8H8)U(BH4)(THF) 2, where Ring = C5H5 2 complex [LU(BH4)][B(C6F5)4] was also in the database borohydrides, such as the mono borohydride [(PC4Me4)2U(BH4)]2 U(BH4)3]2 2(\u03bc-X) where X = O2\u2212 2\u2212 4(\u03bc-L) complexes with Cp\u20323U(BH4) compound was also characterized by 1H, 11B{1H}, 13C{1H}, and 29Si{1H} multi-nuclear NMR spectroscopy. It was of particular inter\u00adest to examine the 29Si{1H} spectrum for comparison with previous studies of silicon-containing paramagnetic uranium complexes and paramagnetically shifted over a range of \u03b4 9.6 to \u221222.6 ppm, in the 1H NMR spectrum, and a 29Si{1H} resonance at \u03b4 \u221257.4 ppm was observed, typical of other tetra\u00advalent uranium complexes \u2212 unit showed considerably more shifting and broadening, resonating at \u03b4 \u221259.5 (\u03bd1/2 = 300\u2005Hz) and 79.6 (\u03bd1/2 = 240\u2005Hz) in the 1H and 11B{1H} spectra, respectively. Since the (BH4)\u2212 ligand exhibited a single 1H NMR resonance whereas two distinct hydride environments are present in the solid state, it appears that the complex is fluxional in solution. This is in line with previous studies were sparged with UHP argon (Praxair) and dried by passage through columns containing a copper(II) oxide oxygen scavenger (Q-5) and mol\u00adecular sieves prior to use or stirred over sodium benzo\u00adphenone ketyl, briefly exposed to vacuum several times to degas and distilled under vacuum. All ethereal solvents were stored over activated 4\u2005\u00c5 mol\u00adecular sieves. Deuterated solvents (Cambridge Isotopes) used for nuclear magnetic resonance (NMR) spectroscopy were dried over sodium benzo\u00adphenone ketyl, degassed by three freeze-pump-thaw cycles, and distilled under vacuum before use. The 1H, 11B{1H}, 13C{1H} and 29Si{1H} NMR spectra were recorded on a GN 500, Cryo 500 or Bruker Avance 600 spectrometer operating at 500.2\u2005MHz, 160.1\u2005MHz, 125.8\u2005MHz, and 99.1\u2005MHz for the 500\u2005MHz spectrometers, respectively, and 600.1\u2005MHz, 192.6\u2005MHz, 150.9\u2005MHz and 119.2\u2005MHz for the 600\u2005MHz spectrometer, respectively, at 298\u2005K unless otherwise stated. The 1H and 13C{1H} NMR spectra were referenced inter\u00adnally to solvent resonances, 11B and 29Si{1H} NMR spectra were referenced externally to BF3(Et2O) and SiMe4, respectively, the 29Si{1H} spectra were acquired using the INEPT pulse sequence. The 15-crown-5 (Aldrich) reagent was dried over activated mol\u00adecular sieves and degassed by three freeze\u2013pump\u2013thaw cycles before use. The NaBH4 (Aldrich) reagent was placed under vacuum (10 \u22123 Torr) for 12\u2005h before use. The following compounds were prepared following literature procedures: KCp\u2032 4 was added to a C7D8 solution of Cp\u20323UI in a J-Young NMR tube, an excess of 15-crown-5 (1 drop) was added and the tube was sealed and removed from the glovebox and vortexed (30\u2005s). The NaBH4 was not fully soluble in C7D8 even in the presence of 15-crown-5. After 18\u2005h, NMR spectroscopy showed complete conversion to Cp\u20323U(BH4). The sample was brought back into the glovebox and the volatiles were removed under reduced pressure. The product was then extracted into Et2O, filtered away from white insoluble solids [presumably Na(15-crown-5)I and excess NaBH4] and the volatiles were removed under reduced pressure to give Cp\u20323U(BH4) as a wine-red solid. 1H NMR : \u03b4 9.7 , \u22122.1 , \u221223.1 , \u221259.8 , 4H); 11B{1H} NMR : \u03b4 79.1 ; 13C{1H} NMR : \u03b4 233.1 (C5H4SiMe3), 214.0 (C5H4SiMe3), 185.6 (C5H4SiMe3), 0.4 (C5H4SiMe3); 29Si{1H} NMR : \u03b4 \u221257.7 ; 1H NMR : \u03b4 9.6 , \u22122.0 , \u221222.6 , \u221259.3 , 4H); 11B{1H} NMR : \u03b4 79.6 ; 13C{1H} NMR : \u03b4 232.0 (C5H4SiMe3), 214.2 (C5H4SiMe3), 186.5 (C5H4SiMe3), 0.6 (C5H4SiMe3); 29Si{1H} NMR : \u03b4 \u221257.4 .Solid NaBH2O (5\u2005mL) solution of KCp\u2032 was added to a pale-green solution of U(BH4)4 , also dissolved in Et2O (5\u2005mL). White solids precipitated (presumably KBH4) as the solution quickly turned orange and then slowly changed to dark red (30\u2005min). After stirring the mixture for an additional 12\u2005h, volatiles were removed under reduced pressure, and the product was extracted into hexane leaving white solids behind (presumably KBH4). Removal of the volatiles under reduced pressure gave Cp\u20323U(BH4) as a dark wine-red solid. X-ray quality crystals were grown from a concentrated ether solution at 253\u2005K.An Ethttps://submission.iucr.org/jtkt/serve/z/Utgd9EjfTrqJVoXA/zz0000/0/.Crystal data, data collection and structure refinement details are summarized in Table\u00a023 moieties, respectively. Methyl torsion angles were not refined but constrained to be staggered. The borohydride H atoms were located from a difference-Fourier map and their positions were freely refined. Uiso(H) values were set to a multiple of Ueq(C/B) with 1.5 for CH3 and BH4 and 1.2 for C\u2014H units, respectively.C\u2014H bond distances were constrained to 0.95\u2005\u00c5 for cyclo\u00adpenta\u00addienyl C\u2014H moieties, and to 0.98\u2005\u00c5 for aliphatic CH10.1107/S2056989021002425/zl5005sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021002425/zl5005Isup3.hklStructure factors: contains datablock(s) I. DOI: 2067867CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Inversion-related chains pack to form layers parallel to the bc plane.The cyclo\u00adhexene ring of the tetra\u00adhydro\u00adiso\u00adquinoline unit is non-planar. The two 4-chloro\u00adphenyl groups extend away from one side of this unit while the hydroxyl and acetyl groups extend away from the opposite side. An intra\u00admolecular O\u2014H\u22efO hydrogen bond fixes the rotational orientation of the acetyl group. In the crystal, N\u2014H\u22efO hydrogen bonds form chains of mol\u00adecules extending along the 28H25Cl2N3O3S, the heterocyclic portion of the tetra\u00adhydro\u00adiso\u00adquinoline unit is planar while the cyclo\u00adhexene ring adopts a twist-boat conformation. The two 4-chloro\u00adphenyl groups extend away from one side of this unit while the hydroxyl and acetyl groups extend away from the opposite side and form an intra\u00admolecular O\u2014H\u22efO hydrogen bond. The crystal packing consists of layers parallel to the bc plane. A Hirshfeld surface analysis of the crystal structure indicates that the most important contributions to the crystal packing are from H\u22efH (37.3%), Cl\u22efH/H\u22efCl (17.6%), O\u22efH/H\u22efO (11.1%), C\u22efH/H\u22efC (10.9%) and N\u22efH/H\u22efN (9.7%) inter\u00adactions.In the title mol\u00adecule, C The conformation of this ring approximates a twist-boat conformation. The best planes through the C10\u2013C15 and C23\u2013C28 rings are inclined to the N1/C5\u2013C9 plane by 76.05\u2005(6) and 74.04\u2005(6)\u00b0, respectively. The acetyl group on C2 is in an equatorial position while the hydroxyl group on C3 is axial and these are syn to one another. The C10\u2013C15 ring attached to C1 is close to equatorial and anti with respect to both other substituents \u2005\u00c5, O3\u22efCg1ii = 3.6287\u2005(11)\u2005\u00c5 and C22\u2014O3\u22efCg1ii = 115.38\u2005(8)\u00b0; symmetry code: (ii) x, y, z; where Cg1 is the centroid of the N1/C5-C9 ring) are also observed in the crystal structure.In the crystal, helical chains extending along the s Table\u00a01. Inversis Table\u00a01. In addiCrystal Explorer 17.5 , Cl\u22efH/H\u22efCl , O\u22efH/H\u22efO , C\u22efH/H\u22efC and N\u22efH/H\u22efN . The other contacts are negligible with individual contributions of less than 2.9% and are given in Table\u00a03Fig.\u00a05et al., 2016et al., 2017et al., 2020H-spiro\u00adisoquinolin]-2(1H)-one -yl]-1,2-di\u00adphenyl\u00adethan-1-ol -6,7-dimeth\u00adoxy-3-(meth\u00adoxy\u00addiphenyl\u00admeth\u00adyl)-1-phenyl-1,2,3,4-tetra\u00adhydro\u00adiso\u00adquinoline and graph-set motifs of the hydrogen-bonding network. In the crystal of DUSVIZ, mol\u00adecules are linked via C\u2014H\u22efO hydrogen bonds. For the major disorder component, these form C(11) chains that propagate parallel to the a axis. In the crystal of AKIVUO, a layer structure with the layers parallel to and (10via weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0(ring) inter\u00adactions. In the crystal structure of ENOCIU, various C\u2014H\u22ef\u03c0 and C\u2014H\u22efO inter\u00adactions link the mol\u00adecules. In the crystal of NIWPAL, the mol\u00adecules are linked by N\u2014H\u22efO inter\u00admolecular hydrogen bonds involving the sulfonamide function to form an infinite two-dimensional network parallel to the (001) plane.In the crystal of NAQRIJ, dimers form through complementary sets of inversion-related O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. These are connected into zigzag chains along the H)-thione, with N-(4-chloro\u00adphen\u00adyl)-2-chloro\u00adacetamide and of anhydrous sodium acetate in pure ethanol (30\u2005ml) for 1\u2005h as shown in Fig.\u00a06The title compound was obtained by refluxing of 7-acetyl-8-(4-chloro\u00adphen\u00adyl)-4-cyano-1,6-dimethyl-6-hy\u00addroxy-5,6,7,8-tetra\u00adhydro\u00adiso\u00adquinoline-3(2\u22121 (O\u2014H), 3277\u2005cm\u22121 (N\u2014H), 2991, 2920\u2005cm\u22121 , 2217\u2005cm\u22121 (C\u2261N), 1694 , 1666\u2005cm\u22121 . 1H NMR : \u03b4 10.95 ; 8.17\u20138.24 ; 7.79\u20137.81 ; 7.26\u20137.32 ; 7.03\u20137.05 ; 4.88 ; 4.53\u20134.55 ; 4.19\u20134.20 ; 3.24\u20133.29 ; 2.87\u20132.90 ; 2.13 ; 1.86 ; 1.27 . Analysis calculated for C28H25Cl2N3O3S (554.47): C 60.65%, H 4.54%, N 7.58%, S 5.78%. Found: C 60.34%, H 4.57%, N 7.68%, S 5.97%.IR: 3522\u2005cmCrystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989021003674/vm2246sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989021003674/vm2246Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021003674/vm2246Isup3.cmlSupporting information file. DOI: 2075592CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The ferrocenyl groups are linked by C\u2014H\u22ef\u03c0(ring) inter\u00adactions.The unsubstituted cyclo\u00adpenta\u00addienyl ring is rotationally disordered while the other Cp ring and its substituent are close to coplanar. In the crystal, the mol\u00adecules pack in \u2018bilayers\u2019 parallel to the 5H5)(C18H13ClN)], is nearly coplanar with the phenyl-1-(4-chloro\u00adphen\u00adyl)methanimine substituent, with dihedral angles between the planes of the phenyl\u00adene ring and the Cp and 4-(chloro\u00adphen\u00adyl)methanimine units of 7.87\u2005(19) and 9.23\u2005(10)\u00b0, respectively. The unsubstituted cyclo\u00adpenta\u00addienyl ring is rotationally disordered, the occupancy ratio for the two orientations refined to a 0.666\u2005(7)/0.334\u2005(7) ratio. In the crystal, the mol\u00adecules pack in \u2018bilayers\u2019 parallel to the ab plane with the ferrocenyl groups on the outer faces and the substituents directed towards the regions between them. The ferrocenyl groups are linked by C\u2014H\u22ef\u03c0(ring) inter\u00adactions. A Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (46.1%), H\u22efC/C\u22ef H (35.4%) and H\u22efCl/Cl\u22efH (13.8%) inter\u00adactions. Thus C\u2014H\u22ef\u03c0(ring) and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing.The substituted cyclo\u00adpenta\u00addienyl ring in the title mol\u00adecule, [Fe(C They are, however, rotated along their long axis with respect to each other, and despite their nearly coplanar nature that predestines them for \u03c0-stacking inter\u00adactions, no such inter\u00adactions are observed in the solid state. Indeed, directional inter\u00adactions are sparse in the structure of the title compound. Ferrocenyl groups are tied together by C\u2014H\u22ef\u03c0 inter\u00adactions, facilitated by neighboring ferrocene units within each layer being roughly 90\u00b0 rotated against each other. Cp-H atoms thus point towards the \u03c0-system of neighboring Cp rings. The shortest C\u2014H\u22ef\u03c0 inter\u00adactions are between H5 and H7 towards the C atoms C7 and C10 of the substituted Cp ring at \u2212x\u00a0+\u00a0y\u00a0+\u00a0z , and between H3 and H10 towards C atoms C4 and C3 at \u2212x\u00a0+\u00a0y\u00a0\u2212\u00a0z . The shortest C\u2014H centroid inter\u00adaction is for C7\u2014H7\u22efCg2 . Also present is a C22\u2014H22\u22efCg5 inter\u00adaction and a weak C4\u2014H4\u22efCl1 hydrogen bond , C\u22efN/N\u22efC and Cl\u22efCl contacts have only 0.5%, 0.2% and 0.1% contributions.In order to visualize the inter\u00admolecular inter\u00adactions in the crystal of the title compound, a Hirshfeld surface (HS) analysis found three closely related, ferrocene-substituted Schiff base compounds: . In a 250\u2005mL round-bottom flask, 1.0\u2005mmol of 4-ferrocenyl aniline in 15\u2005mL of dried methanol was mixed with an equimolar amount of 4-chloro\u00adphenyl aldehyde in 15\u2005mL of dried methanol. The mixture was agitated under reflux, the progress of the reaction was monitored by TLC, and the desired product was formed within 6\u2005h. The solvent was removed under vacuum and the solid that was obtained was recrystallized from methanol (yield: 87%) to yield brown crystals, m.p. 210\u2005K. 1H NMR \u03b4 4.08 ; 4.36 ; 4.68 ; 7.20 ; 7.48 ; 7.53 ; 7.88 ; 8.52 . 13C NMR \u03b4 66.42 ; 69.05 ; 69,64 ; 84.80 ; 121.10 ; 126.76 ; 129.09 ; 129.87 ; 134.92 ; 137.20 ; 137.72 ; 149.21 ; 157.62 .4-Ferrocenyl aniline was synthesized according to a reported procedure (Hu I/\u03c3(I) > 15 and chosen from the full data set with CELL_NOW with a second component (14%) rotated 180\u00b0 about the b axis. To eliminate possible bias, the raw data were processed as triclinic using the multi-component version of SAINT . The data were corrected for absorption using TWINABS . ADPs of equivalent major and minor disordered C atoms were constrained to be identical. The occupancy ratio for the two orientations refined to a 0.666\u2005(7)/0.334\u2005(7) ratio.H atoms attached to carbon were placed in calculated positions (C\u2014H = 0.95\u20131.00\u2005\u00c5). All were included as riding contributions with isotropic displacement parameters 1.2\u20131.5 times those of the parent atoms. The unsubstituted cyclo\u00adpenta\u00addienyl ring is rotationally disordered over two sets of sites with the two components refined as rigid penta\u00adgons I, global. DOI: 10.1107/S2056989021008033/zl5016Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021008033/zl5016Isup3.cdxSupporting information file. DOI: 2101472CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-021-85236-z, published online 02 April 2021Correction to: P values.The original version of this Article contained an error in the legend of Figure\u00a04, where the minus sign was missing in the A) Densities of Alpha for low-stimulus and high-stimulus groups shows 1.23\u2009\u00d7\u2009difference in means with\u00a0P\u2009<\u20092.9E04 (K\u2013S test). (B) Densities for Ea shows 4.8\u2009\u00d7\u2009difference in means with\u00a0P\u2009<\u20093.9E05 (K\u2013S test). (C) Densities for CV_Alpha shows 3.5\u2009\u00d7\u2009difference in means with\u00a0P\u2009<\u20095.8E04 (K\u2013S test). (D) Densities for Theta/Beta ratio shows 1.13\u2009\u00d7\u2009difference in means . (E) Densities for Complexity shows 1.17\u2009\u00d7\u2009difference in means with\u00a0P\u2009<\u20097.5E05 (K\u2013S test). (F) Average power spectrums of low- and high- stimulus groups. Error bars: average spatial variability across individuals.\u201d\u201cDifferences between low-stimulus and high-stimulus groups. (now reads:A) Densities of Alpha for low-stimulus and high-stimulus groups shows 1.23\u2009\u00d7\u2009difference in means with\u00a0P\u2009<\u20092.9E-04 (K\u2013S test). (B) Densities for Ea shows 4.8\u2009\u00d7\u2009difference in means with\u00a0P\u2009<\u20093.9E-05 (K\u2013S test). (C) Densities for CV_Alpha shows 3.5\u2009\u00d7\u2009difference in means with\u00a0P\u2009<\u20095.8E-04 (K\u2013S test). (D) Densities for Theta/Beta ratio shows 1.13\u2009\u00d7\u2009difference in means . (E) Densities for Complexity shows 1.17\u2009\u00d7\u2009difference in means with\u00a0P\u2009<\u20097.5E-05 (K\u2013S test). (F) Average power spectrums of low- and high- stimulus groups. Error bars: average spatial variability across individuals.\u201d\u201cDifferences between low-stimulus and high-stimulus groups. (The original Article has been corrected."} +{"text": "An unusual and labile spirocyclic tetrahedral intermediate (72+) is observed in CO2\u2010pressurized (0.5\u20132.0\u2005bar) solutions of cation 4+ at low temperatures, as demonstrated by variable\u2010temperature NMR studies, which were confirmed crystallographically. In addition, cations 3+ and 4+ actively bind carbonyls, nitriles and acetylenes by 1,3\u2010dipolar cycloaddition, as shown by selected examples.Electrophilic fluorophosphonium triflates bearing pyridyl ( Extending the FLP library: Electrophilic fluorophosphonium triflates bearing pyridyl or imidazolyl substituents act as intramolecular N/P frustrated Lewis pairs that are capable of small molecule activation, that is, reversible CO2 sequestration and binding of carbonyls, nitriles and acetylenes. The corresponding 31P and 19F NMR spectra of the solution containing 3[OTf] reveal no interaction or reaction at ambient temperature indicated by the resonances for cation 3+ (\u03b4(31P)=54.8\u2005ppm, \u03b4(19F)=\u2212114.0\u2005ppm, 1JPF=996\u2005Hz) in the spectra. However, a VT NMR investigation disclosed the reversible and temperature\u2010dependent binding of CO2 by 3[OTf] , a small high field shifted doublet resonance at \u03b4(31P)=\u221255.5\u2005ppm , which is well in the region for a penta\u2010coordinate phosphorus atom, thus, indicating the formation of the CO2 adduct 5[OTf]. The corresponding resonance in the 19F NMR spectrum is detected as a downfield shifted doublet resonance at \u03b4(19F)=\u221230.0\u2005ppm. When the temperature is further decreased to 213\u2005K the integral ratio of 5[OTf] increases to 41\u2009:\u200959. The carbon atom of the bound CO2 is observed as a doublet resonance at \u03b4(13C)=141.8\u2005ppm (2JPC=3\u2005Hz). Further decrease of the temperature to 193\u2005K leads to the separation of a colorless precipitate, suggesting that 5[OTf] exhibits a reduced solubility in CD2Cl2 at low temperature. Accordingly, gradually increase to RT leads to the dissolution of 5[OTf] and its disappearance in the NMR spectrum. This process is reversible without decomposition of 3[OTf]. We investigate this interaction of 3[OTf] with CO2 theoretically with two different models than model 3+\u20102CO_B (\u221218.4\u2005kJ/mol). Moreover, the calculated 31P NMR chemical shift change (\u0394\u03b4=\u03b4(3+\u20102CO_A)\u2212\u03b4(3+)) for model 3+\u20102CO_A (\u0394\u03b4(31P)=\u2212124\u2005ppm) is in the range of the experimental result (\u0394\u03b4(31P)=\u2212110.7\u2005ppm), while the calculated \u0394\u03b4(31P) for model 3+\u20102CO_B is only shifted by \u221217\u2005ppm which is not observed in the respective 31P NMR spectrum. Thus, the CO2 activation is proposed to proceed via the P and pyridyl N atoms, affording 5[OTf] in the configuration of model 3+\u20102CO_A. As the DFT calculation reveals a slightly improved CO2 binding energy (\u221223.0\u2005kJ/mol) for 4[OTf], we also investigated the reactivity of 4[OTf] towards CO2 at ambient temperature results in the immediate formation of a colorless solid. However, the solid re\u2010dissolves immediately after releasing the pressure and all our separation attempts ended up with the recovery of 4[OTf]. We therefore monitored the reaction of 4[OTf] with CO2 (1.5\u2005bar) by NMR spectroscopy in degassed CD3NO2. The 31P NMR spectrum at ambient temperature shows signals of two different adducts at \u03b4(31P)=\u221260.0\u2005ppm and \u03b4(31P)=\u221261.5\u2005ppm besides the signal of 4[OTf] (\u03b4(31P)=55.2\u2005ppm, \u03b4(19F)=\u2212111.5\u2005ppm, 1JPF=1045\u2005Hz, Figure\u200519F spectrum are detected at \u03b4(19F)=\u22128.1\u2005ppm (1JFP=737\u2005Hz) and \u03b4(19F)=\u221216.9\u2005ppm (1JFP=740\u2005Hz), respectively. Interestingly, the 13C NMR spectrum displays two signals for the bound CO2 moiety at \u03b4(13C)=139.9\u2005ppm (2JPC=7\u2005Hz) as a doublet resonance and at \u03b4(13C)=105.5\u2005ppm (2JPC=8\u2005Hz) as a triplet resonance suggesting a coupling to two phosphorus atoms.13 This strongly indicates the formation of a 1\u2009:\u20091 FLP\u2010CO2 adduct 6[OTf] (\u03b4(31P)=\u221261.5\u2005ppm, \u03b4(19F)=\u221216.9\u2005ppm, 1JPF=740\u2005Hz) and a 2\u2009:\u20091 FLP\u2010CO2 adduct 7[OTf]2 (\u03b4(31P)=\u221260.0\u2005ppm, \u03b4(19F)=\u22128.1\u2005ppm, 1JPF=737\u2005Hz). The 31P NMR chemical shift change of 6[OTf] (\u0394\u03b4=\u03b4(6+)\u2212\u03b4(4+), \u2212116.7\u2005ppm) is well in the range of calculated model 4+\u20102CO_A (\u0394\u03b4(31P)=\u2212108\u2005ppm). Decreasing the temperature stepwise to 243\u2005K, the integral ratio of 7[OTf]2 gradually increases to 66\u2009%, while the integral ratio of 6[OTf] increases to 28\u2009%. Variable\u2010temperature NMR studies at 0.5 or 2.0\u2005bar CO2 pressure did not show significant difference on the integral ratio of 6[OTf] and 7[OTf]2. Although the calculated electrophilicities are similar for 3[OTf] and 4[OTf] ,2 with 3[OTf]. This suggests that a certain nucleophilicity of the Lewis basic site is crucial for the formation of the 2\u2009:\u20091 FLP\u2010CO2 adduct 7[OTf]2.The 6+ and 72+ by vapor diffusion of CH2Cl2 into a CH3NO2 solution of 4[OTf] under CO2 atmosphere (0.5\u2005bar) at \u221230\u2009\u00b0C and aluminum (VII) complexes \u00b0. One oxygen is bound by the Lewis acidic P atom with a P1\u2212O1 bond length of 1.773(5)\u2005\u00c5 and the carbon atom is stabilized by the Lewis basic N atom to give a N1\u2212C1 bond length of 1.442(10)\u2005\u00c5. The C1\u2212O1 (1.307(8)\u2005\u00c5) and C1=O2 (1.206(10)\u2005\u00c5) bonds in the CO2 fragment are comparable to other FLP\u2010CO2 adducts . (+9\u2009a/+9\u2009b: \u03b4(31P)=\u221249.3/\u221265.0\u2005ppm, \u03b4(19F)=3.0/19.9\u2005ppm, 1JPF=738/702\u2005Hz, 71 and 95\u2009% isolated yield) are indicated by multinuclear NMR spectroscopy =101.8/96.7\u2005ppm, 2JCP=9/11\u2005Hz) are significantly shifted to higher field compared to the free acetone (\u03b4(13C)=207\u2005ppm).9\u2009a,b[OTf] are shifted to lower field (9\u2009a+/9\u2009b+: \u03b4(13C)=151.0/150.2\u2005ppm, 2JCP=18/19\u2005Hz). Interestingly, when compounds 9\u2009a,\u2009b[OTf] are reacted with acetone at room temperature, the formation of 8\u2009a,\u2009b[OTf] under release of CH3CN is observed after a reaction time of 1\u2005h at room temperature for 4\u2005h, the formation of 10\u2009a,\u2009b and 11\u2009a,\u2009b[OTf]2 is observed, while 12\u2009a,\u2009b[OTf] are cleanly formed by fluoride abstraction from 10\u2009a,\u2009b to 11\u2009a,\u2009b2+ after reacting at 80\u2009\u00b0C overnight or at ambient temperature for 2 weeks is more Lewis acidic than 3+ , thus, low reaction tendency from 11\u2009a,\u2009b+ to 12\u2009a,\u2009b+ seems to be influenced by solvent effects, which significantly decrease the Lewis acidity compared to the gas phase calculations.2 are found as singlet resonances in the 1P NMR spectra at \u03b4(31P)=6.9 and \u221215.0\u2005ppm, while 12\u2009a,\u2009b[OTf] are observed as doublet resonances in the 31P and 19F NMR spectra (12\u2009a+/12\u2009b+: \u03b4(31P)=\u221288.8/\u221298.8\u2005ppm, \u03b4(19F)=34.0/25.2\u2005ppm, 1JPF=707/718\u2005Hz, Table\u200511\u2009a[OTf]212\u2009b[OTf] (66\u2009%) as pure products for full characterization.When s Scheme\u2005, iii\u2013v. 8\u2009a,\u2009b[OTf], 9\u2009a,\u2009b[OTf] and 12\u2009b[OTf] are confirmed by X\u2010ray analysis of suitable single crystals which were obtained by slow vapor diffusion of n\u2010pentane into saturated CH2Cl2 solutions at \u221230\u2009\u00b0C and the fluorine atoms occupy the axial position. The P\u2212F bonds (1.637(2) \u22121.673(13)\u2005\u00c5) are elongated compared to those of the starting materials 3+ and 4+, but comparable to those in difluorophosphoranes 3PF2: 1.638(2)\u2005\u00c5).8\u2009a+/8\u2009b+: 1.695(2)/1.695(3)\u2005\u00c5) are significantly shorter compared to the acetonitrile adducts (9\u2009a+/9\u2009b+: 1.777(2)/1.788(2)\u2005\u00c5) while the C\u2212N bonds are around 0.15\u2005\u00c5 longer (8\u2009a+/8\u2009b+: 1.495(4)\u2005\u00c5/1.411(5)\u2005\u00c5, 9\u2009a+/9\u2009b+: 1.250(3)/1.272(3)\u2005\u00c5, Table\u2005The molecular structures of compounds C Figure\u2005. All obt8\u2009a+/8\u2009b+: 1.495(4)/1.411(5)\u2005\u00c5 and the C\u2212N distances of the nitrile moiety (9\u2009a+/9\u2009b+: 1.250(3)/1.272(3)\u2005\u00c5) are typical for C\u2212O single bonds and C=N double bonds, respectively.12\u2009b+ shows that the pyridyl moiety is in the axial position, opposite to the fluorine atom , 2061971 (for 2[OTf]), 2061972 (for 3[OTf]\u2009\u22c5\u2009C6H5F), 2061973 (for 6[OTf]\u2009\u22c5\u20097[OTf]2\u2009\u22c5\u2009CH3NO2), 2061974 (for 8\u2009a[OTf]\u2009\u22c5\u2009CH2Cl2), 2061975 (for 8\u2009b[OTf]\u2009\u22c5\u2009CH2Cl2), 2061976 (for 9\u2009a[OTf]\u2009\u22c5\u2009(CH2Cl2)2\u2009\u22c5\u2009CH3CN), 2061977 (for 9\u2009b[OTf]), 2061978 (for 13), 2061979 (for 12b[OTf]), and 2061980 (for 14)2061970 or imidazolyl (4[OTf]) substituents, thus an additional Lewis basic site, represent a new type of intramolecular FLP that reacts with small molecules in a cooperative manner. The cyclizations of 3[OTf] and 4[OTf] with the 1,2\u2010dipolar compounds acetone, acetonitrile and acetylenes gave rise to the formation of the corresponding heterocyclic compounds, thus illustrating the cooperative reactivity of the new FLP derivatives. This is additionally demonstrated by the reversible formation of adducts with CO2 (5+ and 6+) at low temperature, which was investigated by variable\u2010temperature NMR studies and X\u2010ray analysis. Surprisingly, the bifunctional phosphonium cation 4+ forms an adduct with CO2 (72+) comprising one molecule of CO2 and two molecules of 4+, resulting in a spirocyclic geometry at the central carbon atom, which is hitherto unreported for an N/P FLP system. These novel bifunctional phosphonium cations extend the diverse library of FLP systems, and the reversible CO2 adduct formation might provide new applications for in FLP chemistry, which we are currently investigating.In summary, electrophilic fluorophosphonium compounds bearing pyridyl should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "In the crystal, chains parallel to the c axis are generated by inter\u00admolecular N\u2014H\u22efO hydrogen bonds with the chains assembled into a three-dimensional network structure by inter\u00admolecular C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0(ring) inter\u00adactions. The mol\u00adecule of (II), C22H21N3O, differs from (I) only in the substituent at the hydrazide N atom where a phenyl\u00admethyl\u00adene moiety for (II) is present instead of a furan\u00admethyl\u00adene moiety for (I). Hence, mol\u00adecules of (I) and (II) show similarities in their mol\u00adecular and crystal structures. The conformation of the central portion of the mol\u00adecule of (II) is also therefore partially determined by an intra\u00admolecular N\u2014H\u22efO hydrogen bond and inter\u00admolecular N\u2014H\u22efO hydrogen bonds form chains parallel to the c axis. Likewise, the chains are connected into a three-dimensional network by C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0(ring) inter\u00adactions.The conformation about the central benzene ring in the mol\u00adecule of (I), C These cs Table\u00a01.c-axis direction n Table\u00a02, which ak Table\u00a02.et al., 2016N\u2032-methyl\u00adidene\u00adbenzohydrazide skeleton: N\u2032-[(4-chloro\u00adphen\u00adyl)methyl\u00adidene]-2-[amino]\u00adbenzohydrazide ethyl\u00adidene]-2-[amino]\u00adbenzohydrazide benzohydrazide benzohydrazide -2-[amino]-N\u2032-(2-methyl-5-(prop-1-en-2-yl)cyclo\u00adhex-2-en-1-yl\u00adidene)benzo\u00adhydra\u00adzide \u00b0. These rings make dihedral angles of 47.79\u2005(8) and 69.24\u2005(9)\u00b0, respectively, with the central benzene ring. In the crystal structure of VEDBAK, mol\u00adecules are linked into a three-dimensional supra\u00admolecular network by N\u2014H\u22efO, C\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22ef\u03c0 inter\u00adactions.In the crystal structure of LEBSET, mol\u00adecules are linked into a three-dimensional supra\u00admolecular network by N\u2014H\u22efN, N\u2014H\u22efO, C\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22ef\u03c0 inter\u00adactions.A and B) having differing conformations that mainly concern the dihedral angles between the hy\u00addroxy\u00adphenyl and di\u00admethyl\u00adphenyl rings relative to the central phenyl\u00adene ring, with values of 30.16\u2005(6) and 58.60\u2005(6)\u00b0 in mol\u00adecule A and of 13.42\u2005(7) and 60.31\u2005(7)\u00b0 in mol\u00adecule B. With the exception of the di\u00admethyl\u00adphenyl substituent, the conformations of the rest of each mol\u00adecule are largely determined by intra\u00admolecular O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds. In the crystal structure, N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into chains extending parallel to the a axis where the types of mol\u00adecules alternate in an \u22efA\u22efB\u22efA\u22efB\u22ef fashion.The asymmetric unit of DABREG consists of two mol\u00adecules (S(6) ring motif. In the crystal structure of LEGHAI, mol\u00adecules are linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds to the same acceptor atom, forming sheets lying parallel to the bc plane. The crystal packing also features C\u2014H\u22ef\u03c0 inter\u00adactions.In LEGHAI, the central benzene ring makes dihedral angles of 45.36\u2005(9) and 55.33\u2005(9)\u00b0 with the thio\u00adphene ring and the dimethyl-substituted benzene ring, respectively. The dihedral angle between the thio\u00adphene ring and dimethyl-substituted benzene ring is 83.60\u2005(9)\u00b0. The thio\u00adphene ring and the benzene ring are twisted from the mean plane of the C(=O)\u2014N\u2014N=C bridge [maximum deviation = 0.0860\u2005(13)\u2005\u00c5], with dihedral angles of 23.86\u2005(9) and 24.77\u2005(8)\u00b0, respectively. An intra\u00admolecular N\u2014H\u22efO hydrogen bond generates an S(6) ring motif in the mol\u00adecule, and a short intra\u00admolecular contact (H\u22efH = 1.88\u2005\u00c5) is also observed. In the crystal structure of LEGHIQ, mol\u00adecules are linked by pairs of N\u2014H\u22efN hydrogen bonds into inversion dimers. The crystal packing also features C\u2014H\u22ef\u03c0 inter\u00adactions.In LEGHIQ, the dihedral angle between the benzene rings is 58.05\u2005(9)\u00b0. The non-H atoms of the hydrazide group lie in a common plane (r.m.s. deviation = 0.0006\u2005\u00c5) and are close to co-planar with their attached benzene ring [dihedral angle = 8.02\u2005(9)\u00b0]. An intra\u00admolecular N\u2014H\u22efO hydrogen bond generates an via N\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonds, forming chains along the a-axis direction. In each mol\u00adecule, there is an intra\u00admolecular N\u2014H\u22efO hydrogen bond.The asymmetric unit of the compound YAXJUE comprises two mol\u00adecules. The dihedral angles between the benzene rings in the two mol\u00adecules are 59.7\u2005(2) and 61.27\u2005(18)\u00b0. The cyclo\u00adhexene rings adopt sofa and half-chair conformations. In the crystal structure of YAXJUE, mol\u00adecules are connected Synthesis of (I) A mixture of 1\u2005mmol of 2-furaldehyde (96\u2005mg) and 1\u2005mmol of 2-[amino]\u00adbenzohydrazide (255\u2005mg) in 20\u2005ml of ethanol was refluxed and monitored by TLC until completion. The reaction mixture was cooled to room temperature when the solid product was obtained. The crude product was filtered off, dried and recrystallized from ethanol to afford crystals suitable for X-ray diffraction. M.p. 479\u2013483\u2005K.Synthesis of (II) In a solution of 20\u2005ml of ethanol, a mixture of 106\u2005mg (1\u2005mmol) of benzaldehyde (106\u2005mg) and 255\u2005mg (1\u2005mmol) of 2-[amino]\u00adbenzohydrazide was refluxed for 4\u2005h. The solid product was obtained after the reaction mixture was cooled to room temperature. The crude product was filtered off, dried and recrystallized from ethanol to afford crystals suitable for X-ray diffraction. M.p. 466\u2013469\u2005K.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021001353/wm5599sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989021001353/wm5599Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989021001353/wm5599IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989021001353/wm5599Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021001353/wm5599IIsup5.cmlSupporting information file. DOI: 2061393, 2061392CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The -2,2\u2032-{[bis\u00ad[(aza\u00adniumylyl\u00adidene)methanylyl\u00adidene]}bis\u00ad(6-meth\u00adoxy\u00adphenolate) ligand, obtained from o-vanillin and -(\u2212)-1,2-di\u00adphenyl\u00adethyl\u00adenedi\u00adamine, exhibits a slightly distorted planar arrangement of the four coordinated O atoms. In the crystal, the complex shows intra\u00admolecular N\u2014H\u22efO hydrogen bonds and weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds. The Hirshfeld surface analysis indicates that the most important contributions to the packing are from H\u22efH (33.5%), O\u22efH (34.1%) and C\u22efH (21.7%) contacts.In the title complex, [Sm(NO The bond lengths between the metal center and the nitrate oxygen atoms range from 2.475\u2005(5) to 2.633\u2005(5)\u2005\u00c5, showing more flexibility than those of the Schiff base ligand. In the Schiff base ligand, the imine moieties are protonated to form iminium cations, but the C=N bond lengths remain close to those of normal imine bonds at 1.287\u2005(8) and 1.30\u2005(1)\u2005\u00c5.et al., 2013Intra\u00admolecular hydrogen bonds occur between the iminium protons and the phenolic oxygen atoms, with lengths of 1.71\u20131.89\u2005\u00c5 Table\u00a01. The bonet al., 2009et al., 2004CrystalExplorer17.5 -3-(meth\u00adoxy)phen\u00adyl]methaniminiumato})tris(nitrato)samarium -{\u03bc-}bis[6-(meth\u00adoxy)phenolato]]}trinitratoeuropium(III)nickel(II) -(\u2212)-1,2-Di\u00adphenyl\u00adethyl\u00adenedi\u00adamine and o-vanillin were dissolved in ethanol (30\u2005mL) and the resulting mixture was stirred at 313\u2005K for 1\u2005h to afford a yellow solution. To this solution, samarium nitrate hexa\u00adhydrate was added and it was stirred at 313\u2005K for 2\u2005h. A yellow precipitate appeared immediately. The precipitate was filtered and washed with ethanol and hexane. The title compound was obtained as a yellow solid. IR : 1624 (C=N double bond). Fluorescence bands in methanol solution were observed at 562 (4G5/2 \u2192 6H5/2), 597 (4G5/2 \u2192 6H7/2) and 644 (4G5/2 \u2192 6H 9/2) nm. Single crystals suitable for X-ray diffraction were obtained by recrystallization from methanol and diethyl ether solution.(1Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl). SIMU, ISOR and AFIX 66 commands were used for C55, C56, C57, C58, C59, C60 to suppress temperature anisotropy and restrain bond lengths to appropriate values.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021004424/tx2039sup1.cifCrystal structure: contains datablock(s) 1R, I. DOI: 10.1107/S2056989021004424/tx2039Isup2.hklStructure factors: contains datablock(s) I. DOI: 2080014CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-020-72306-x, published online 17 September 2020Correction to: The original version of this Article contained an error.The p value stated in the text for differences in fasting blood sugar, between non-overweight and obese groups, was incorrect. The text,\u201cThere was no statistically significant difference in the level of ALT (98.9\u2009\u00b1\u200934.6 vs. 96.3\u2009\u00b1\u200924.1 p\u2009=\u20090.76), serum vitamin B12 (559\u2009\u00b1\u2009417.1 vs. 433\u2009\u00b1\u2009185.4 p\u2009=\u20090.18), fasting blood sugar or the serum homocysteine levels (13.2\u2009\u00b1\u20094.0 vs. 14.04\u2009+\u20091.80 p\u2009=\u20090.49) between the non-overweight and obese groups.\u201dnow reads,\u201cThere was no statistically significant difference in the level of ALT (98.9\u2009\u00b1\u200934.6 vs. 96.3\u2009\u00b1\u200924.1 p\u2009=\u20090.76), serum vitamin B12 (559\u2009\u00b1\u2009417.1 vs. 433\u2009\u00b1\u2009185.4 p\u2009=\u20090.18) or the serum homocysteine levels (13.2\u2009\u00b1\u20094.0 vs. 14.04\u2009+\u20091.80 p\u2009=\u20090.49) between the non-overweight and obese groups. Though the fasting blood sugar was significantly different, there was only one patient whose fasting blood sugar placed them within range of a diabetes mellitus diagnosis .\u201dThis has been corrected in the PDF and HTML versions of the Article."} +{"text": "Thereby, each signal alone only induces a moderate production of pro\u2010inflammatory cytokines and lipid mediators (LMs). However, MCs, which simultaneously sense (co\u2010sensing) IL\u201033 and ATP, display an enhanced and prolonged activation of the TAK1\u2010IKK2\u2010NF\u2010\u03baB signalling pathway. This resulted in a massive production of pro\u2010inflammatory cytokines such as IL\u20102, IL\u20104, IL\u20106 and GM\u2010CSF as well as of arachidonic acid\u2010derived cyclooxygenase (COX)\u2010mediated pro\u2010inflammatory prostaglandins (PGs) and thromboxanes (TXs), hallmarks of strong MC activation. Collectively, these data show that co\u2010sensing of ATP and IL\u201033 results in hyperactivation of MCs, which resembles to MC activation induced by IgE\u2010mediated crosslinking of the Fc\u03b5RI. Therefore, the IL\u201033/IL\u201033R and/or the ATP/P2X7 signalling axis are attractive targets for therapeutical intervention of diseases associated with the loss of integrity of cellular barriers such as allergic and infectious respiratory reactions.IL\u201033 and ATP are alarmins, which are released upon damage of cellular barriers or are actively secreted upon cell stress. Due to high\u2010density expression of the IL\u201033 receptor T1/ST2 (IL\u201033R), and the ATP receptor P2X7, mast cells (MCs) are one of the first highly sensitive sentinels recognizing released IL\u201033 or ATP in damaged peripheral tissues. Whereas IL\u201033 induces the MyD88\u2010dependent activation of the TAK1\u2010IKK2\u2010NF\u2010\u03baB signalling, ATP induces the Ca Co\u2010sensing of ATP and IL\u201033 triggers hyperactivation of mast cells resulting in an amplified production of pro\u2010inflammatory cytokines and eicosanoids. Imagewas created with BioRender.com. After centrifugation, cells were recovered in KREBS\u2010HEPES buffer supplemented with 1\u00a0mM CaCl2 and diluted to a final density of 1\u00a0\u00d7\u00a0106 cells/ml. Cells were transferred onto black 96\u2010well microplates PS F\u2010bottom (Greiner bio\u2010one), and the signal was monitored by a NOVOstar microplate reader (BMG Labtechnologies GmbH) at 37\u00b0C : emission at 510\u00a0nm, excitation at 340\u00a0nm (Ca2+\u2010bound Fura\u20102) and 380\u00a0nm (free Fura\u20102). By subsequent cell lysis with triton X\u2010100, the maximal fluorescence signals were determined, followed by chelating Ca2+ with 20\u00a0mM EDTA to assess the minimal fluorescence.To determine the effect of ATP (Sigma) and/ or IL\u201033 (50\u00a0ng/ml) (PeproTech) on the intracellular Ca3). BMMCs were treated with anti\u2010CD16/CD32 and rat\u2010IgG (Jackson) to prevent non\u2010specific binding of the staining antibodies. BMMCs were stained with PE\u2010anti\u2010CD117 or APC\u2010anti\u2010CD117, FITC\u2010anti\u2010Fc\u03b5RI or PE\u2010CD107\u03b1 . MC/9\u2010NFAT and MC/9\u2010NF\u2010\u03baB MC lines were stained with PE\u2010anti\u2010CD117. All cells were analysed with a LSR II flow cytometer (BD). Data were analysed with FlowJo 10 .BMMCs were harvested and washed with PBA (PeproTech) and/or ATP (500\u00a0\u00b5M) (Sigma). Lysis of the cells was performed with lysis buffer , and the protein concentration was determined . Boiled samples (treated with 6\u00a0\u00d7\u00a0Laemmli buffer) were separated on 10% sodium dodecyl sulphate (SDS)\u2010Laemmli gels. Gels were transferred onto nitrocellulose membranes (biostep) by electroblotting. After blocking (with dry milk), membranes were incubated with either anti\u2010pT180/pY182\u2010p38, anti\u2010pS177/181\u2010IKK2, anti\u2010p184/187\u2010pTAK1, anti\u2010pT202/pY204\u2010ERK1/2, anti\u2010pS32\u2010I\u03baB\u03b1, and anti\u2010pS536\u2010p65 or the respective anti\u2010total antibodies (non\u2010phosphorylated proteins) . Washed membranes (TBS/0\u00b71% Tween) were incubated with HRP\u2010conjugated secondary anti\u2010rabbit\u2010Ig, or anti\u2010mouse\u2010Ig or anti\u2010goat\u2010Ig (SeraCare). Detection was performed with ECL reagent (Pierce).Washed BMMCs were seeded in IL\u20103\u2010free media at a density of 106 cells/ml) were seeded in IL\u20103\u2010free media. After 1\u00a0h, cells were pre\u2010treated with the inhibitors cyclosporine (4\u00a0\u00b5g/ml) , the Ca2+ chelator BAPTA\u2010AM (5\u00a0\u00b5M) (Ca2+ chel.) for Ca2+ depletion, U0126 (10\u00a0\u00b5M) , the selective P2X7 inhibitor, A438079 (50\u00a0\u00b5M) (Sigma) or skepinone\u2010L (1\u00a0\u00b5M) (Cayman) or vehicle for 30\u00a0min. Afterwards, BMMCs were stimulated with IL\u201033 (50\u00a0ng/ml) (PeproTech) or ATP (500\u00a0\u00b5M) (Sigma) or both in combination for 24\u00a0h. Supernatants were transferred to ice\u2010cold methanol containing deuterium\u2010labelled internal standards for the quantification of LM and sample recovery. Sample preparation was performed as recently shown recently reported [2O was added . Samples were subjected to solid\u2010phase extraction using Sep\u2010Pak\u00ae Vac 6cc 500\u00a0mg/6\u00a0ml C18 (Waters). After washing with H2O and n\u2010hexane, LM were eluted with methyl formate. Samples were dried using an evaporation system (TurboVap LV) and resuspended in methanol\u2010water for UPLC\u2010MS\u2010MS automated injections. LM profiling was analysed with an Acquity\u2122 UPLC system (Waters) and a QTRAP 5500\u00a0mass spectrometer (ABSciex) equipped with a TurboV\u2122 Source and electrospray ionization. LM was eluted using an ACQUITY UPLC\u00ae BEH C18 column at 50\u00b0C with a flow rate of 0\u00b73\u00a0ml/min and a mobile phase consisting of methanol\u2010water\u2010acetic acid of 42:58:0\u00b701 (v/v/v) that was ramped to 86:14:0\u00b701 (v/v/v) over 12.5\u00a0min and then to 98:2:0\u00b701 (v/v/v) for 3\u00a0min [BMMCs (1\u00a0\u00d7\u00a010reported . In brieor 3\u00a0min . The QTr6 cells/ml) were seeded in IL\u20103\u2010 and serum\u2010free media. After 1\u00a0h, cells were pre\u2010treated with the inhibitors cyclosporine A (4\u00a0\u00b5g/ml) , the Ca2+ chelator BAPTA\u2010AM (5\u00a0\u00b5M) (Ca2+ chel.) for Ca2+ depletion, U0126 (10\u00a0\u00b5M) or skepinone\u2010L (1\u00a0\u00b5M) (Cayman) or vehicle for 30\u00a0min. Afterwards, BMMCs were stimulated with IL\u201033 (50\u00a0ng/ml) (PeproTech) or ATP (500\u00a0\u00b5M) (Sigma) or both in combination for 24\u00a0h. Supernatants were transferred to pre\u2010coated ELISA plates and the competitive LTC4 ELISA was performed as described in the instructions (Cayman).BMMCs . Statistics were performed with IBM SPSS Statistics version 20.0 .All experiments were performed at least with three biological replicates. One biological replicate always is the bone marrow culture from at least 2 mice. Western blots intensities were determined with the ImageJ software (Fiji). The phospho\u2010specific Western blots were normalized to the total protein blots. The control (unstimulated sample) of wt BMMCs was set as 1. For analysis of CaThe authors declare no competing financial interests.P.M.J., N.A., M.G., O.W. and A. D. performed and analysed experiments, wrote and edited the manuscript; P.W., F.W., U.J. and C.K.: performed and analysed experiments, edited the manuscript; T.K. and E.S. provided material and edited the manuscript; S.D. developed the concept, designed the research, performed most of the experiments, analysed data, drafted, wrote and edited the paper.Fig S1Click here for additional data file.Fig S2Click here for additional data file.Fig S3Click here for additional data file.Fig S4Click here for additional data file.Fig S5Click here for additional data file.Fig S6Click here for additional data file.Fig S7Click here for additional data file.Fig S8Click here for additional data file.Fig S9Click here for additional data file."} +{"text": "C\u2014Br\u22ef\u03c0 and C=O\u22ef\u03c0 inter\u00adactions stabilize the mol\u00adecular packing, resulting in a three-dimensional network.In the crystal, N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into dimers with an 20H16BrN3O2, was determined using an inversion twin. Its asymmetric unit comprises two crystallographically independent mol\u00adecules (A and B) being the stereoisomers. Both mol\u00adecules are linked by pairs of N\u2014H\u22efO hydrogen bonds, forming a dimer with an R22(16) ring motif. The dimers are connected by further N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, forming chains along the c-axis direction\u00b7C\u2014Br\u22ef\u03c0 inter\u00adactions between these chains contribute to the stabilization of the mol\u00adecular packing. Hirshfeld surface analysis showed that the most important contributions to the crystal packing are from H\u22efH, C\u22efH/H\u22efC, O\u22efH/H\u22efO, Br\u22efH/H\u22efBr and N\u22efH/H\u22efN inter\u00adactions.The crystal structure of the title compound, C In mol\u00adecule A, the phenyl (C7\u2013C12) and bromo\u00adphenyl (C14\u2013C19) rings form dihedral angles of 64.0\u2005(4) and 86.3\u2005(4)\u00b0, respectively, with the mean plane of the central di\u00adhydro\u00adpyridine ring. In mol\u00adecule B, the corresponding dihedral angles are 77.2\u2005(4) and 83.9\u2005(4)\u00b0. The acetyl groups in both mol\u00adecules are almost planar and they make the dihedral angles of 89.5\u2005(5) and 87.7\u2005(5)\u00b0 with the mean planes of the di\u00adhydro\u00adpyridine rings in these mol\u00adecules.The title compound crystallizes in the monoclinic space group it Fig.\u00a01. These mB\u22efO21 and N26\u2014H26A\u22efO1 hydrogen bonds \u2005\u00c5, C21=O21\u22efCg2 = 110.8\u2005(6)\u00b0, O1\u22efCg5 = 3.748\u2005(8)\u2005\u00c5, C1=O1\u22efCg5 = 125.1\u2005(6)\u00b0, where Cg2 and Cg5 are the centroids of the C7\u2013C12 phenyl ring in mol\u00adecule A and the C27\u2013C32 phenyl ring in mol\u00adecule B, respectively]. The dimers are connected by N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds with an c-axis direction (Table\u00a01Cg6v = 3.407\u2005(4)\u2005\u00c5, C17\u2014Br1\u22efCg6v = 145.2\u2005(3)\u00b0; symmetry code (v) \u22121\u00a0+\u00a0x, 1\u00a0\u2212\u00a0y, \u2212z; Cg6 is the centroid of the C34\u2013C39 ring]. Together with the other inter\u00admolecular contacts given in Table\u00a02Strong N6\u2014H6ds Fig.\u00a01 link moln Table\u00a01 and 5 \u25b8.A and B, CrystalExplorer17 indicate regions of N\u2014H\u22efO inter\u00adactions. The N\u2014H\u22efN and C\u2014H\u22efN inter\u00adactions , the mol\u00adecular conformation of the title compound is stabilized by an intra\u00admolecular O\u2014H\u22efO hydrogen bond, forming an S(6) ring motif. In the crystal, mol\u00adecules are linked by inter\u00admolecular N\u2014H\u22efN and C\u2014H\u22efN hydrogen bonds, and N\u2014H\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional network.In OZAKOS (space group: PPi.e. R32(8)] is formed involving the sulfonate group, the pyrimidinium cation and the water mol\u00adecule. Both salts form a supra\u00admolecular homosynthon [In both the related salts, JEBREQ (space group: P21/n) and SETWOE (space group: P21/c), adopt nearly planar structures. The crystal structure of SETWUK is stabilized by inter\u00admolecular C\u2014H\u22efF and C\u2014H\u22ef\u03c0 inter\u00adactions. The C\u2014H\u22efF bond generates a linear chain with a C(14) motif. The crystal structure of SETWOE is stabilized by inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. The C\u2014H\u22efO hydrogen bonds generate rings with 22R(14) and 22R(20) motifs. In addition, in SETWOE and SETWUK, intra\u00admolecular O\u2014H\u22efO inter\u00adactions are found, which generate an S(6) graph-set motif. No significant ar\u00adyl\u2013aryl or \u03c0\u2013\u03c0 inter\u00adactions exist in these structures. All this bears some resemblance to the title compound.The polysubstituted pyridines, SETWUK (space group: To a solution of 2-(4-bromo\u00adbenzyl\u00adidene)malono\u00adnitrile and acetoacetanilide in methanol (25\u2005mL), piperidine (2\u20133 drops) was added and the mixture was stirred at room temperature for 48\u2005h. Then 15\u2005mL of methanol were removed by rotary evaporation from the reaction mixture, which was left overnight. The precipitated crystals were separated by filtration and recrystallized from ethanol/water (1:1) solution .1H NMR : 2.29 ; 4.15 ; 4.34 ; 5.98 ; 7.12\u20137.35 ; 7.40 ; 7.61 .13C NMR : 27.86 (CH3\u2014C=O), 37.94 (CH\u2014Ar), 57.24 (=Cquat), 62.41 (CH\u2014C=O), 117.21 (CN), 121.25 (Br-Car), 127.67 (CHar), 128.19 (2CHar), 129.58 (2CHar), 130.15 (2CHar), 130.74 (2CHar), 136.98 (Car), 140.37 (Car), 154.14 (=Cquat), 166.20 (N\u2014C=O), 202.55 (C=O).Uiso(H) = 1.2Ueq or 1.5Ueq(C-meth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989022001232/yk2165sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022001232/yk2165Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022001232/yk2165Isup3.cmlSupporting information file. DOI: 2149629CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structures of two triclinic polymorphs of a new tetra\u00adkis-substituted pyrazine carb\u00adoxy\u00adlic acid, 3,3\u2032,3\u2032\u2032,3\u2032\u2032\u2032-[)tetra\u00adkis\u00ad(sulfanedi\u00adyl)]tetra\u00adpropionic acid, are reported, together with the crystal structures of two potassium-organic frameworks. 20H28N2O8S4, (4L1H), have been obtained, 4L1_AH and 4L1_BH. Each structure crystallized with half a mol\u00adecule in the asymmetric unit of a triclinic P4L1_BH were of poor quality, but the X-ray diffraction analysis does show the change in conformation of the \u2013CH2\u2014S\u2014CH2\u2014CH2\u2013 side chains compared to those in polymorph 4L1_AH. In the crystal of 4L1_AH, mol\u00adecules are linked by two pairs of O\u2014H\u22efO hydrogen bonds, enclosing R22(8) ring motifs forming layers parallel to plane (100), which are linked by C\u2014H\u22efO hydrogen bonds to form a supra\u00admolecular framework. In the crystal of 4L1_BH, mol\u00adecules are also linked by two pairs of O\u2014H\u22efO hydrogen bonds enclosing R22(8) ring motifs, however here, chains are formed propagating in the [001] direction and stacking up the a-axis. Reaction of 4L1H with Hg(NO3)2 in the presence of a potassium acetate buffer did not produce the expected binuclear complex, instead crystals of a potassium\u2013organic framework were obtained, poly[sulfan\u00adyl]meth\u00adyl}pyrazin-2-yl)meth\u00adyl]sulfan\u00adyl}propano\u00adato)potassium], [K(C20H27N2O8S4)]n (3L1KH). The organic mono-anion possesses inversion symmetry with the pyrazine ring being located about an inversion center. A carb\u00adoxy H atom is disordered by symmetry and the charge is compensated for by a potassium ion. A similar reaction with Zn(NO3)2 resulted in the formation of crystals of a dipotassium-organic framework, poly[sulfan\u00adyl]meth\u00adyl}pyrazine-2,5-di\u00adyl)bis(methyl\u00adene)]bis\u00ad(sulfanedi\u00adyl)}dipropionato)dipotassium], [K2(C20H26N2O8S4)]n (2H2L1K). Here, the organic di-anion possesses inversion symmetry with the pyrazine ring being located about an inversion center. Two symmetry-related acid groups are deprotonated and the charges are compensated for by two potassium ions.Two polymorphs of the title tetra\u00adkis-substituted pyrazine carb\u00adoxy\u00adlic acid, 3,3\u2032,3\u2032\u2032,3\u2032\u2032\u2032-{}tetra\u00adpropionic acid, C The first is pyrazine-2,3,5,6-tetra\u00adcarb\u00adoxy\u00adlic acid (pztca), which was originally synthesized by Wolff at the end of the 19th century , which was first synthesized by Jiang et al. )tetra\u00adkis\u00ad(sulfane\u00addi\u00adyl)]tetra\u00adpropionic acid , for which two nickel(II) binuclear complexes, I and II, were synthesized , crystallized with half a mol\u00adecule in the asymmetric unit )tetra\u00adkis\u00ad(sulfanedi\u00adyl)]tetra\u00adpropionic acid of the centrosymmetric mol\u00adecule being positionally disordered . The difference in the two polymorphs is essentially in the orientation of the \u2013CH2\u2014S\u2014CH2\u2014CH2\u2014C\u2013 side arms, as shown in Fig.\u00a05a and b. Selected torsion angles are given in Table\u00a01In an attempt to form a co-crystal, equimolar amounts of _B Fig.\u00a04. It crysed Fig.\u00a04b. The d4L1H with Hg(NO3)2 in the presence a 1 M potassium acetate buffer led to the formation of colourless crystals that proved to be a potassium\u2013organic framework (3L1KH); see Fig.\u00a06+ ion is linked to the O atoms of the acid groups and has a coordination number of eight (KO8) and a distorted dodeca\u00adhedral geometry . The K\u22efO bond lengths vary between 2.682\u2005(2) and 3.069\u2005(3)\u2005\u00c5 (Table\u00a02\u2212) distances: 2.6823\u2005(2) and 2.828\u2005(2)\u2005\u00c5 compared to 3.056\u2005(3) and 3.069\u2005(3)\u2005\u00c5, respectively.Reaction of ry Fig.\u00a07a. The K\u00c5 Table\u00a02. Inter\u00ade4L1H with Zn(NO3)2 in the presence of a 1 M potassium acetate buffer led to the formation of colourless crystals that proved to be a dipotassium\u2013organic framework (2H2L1K); see Fig.\u00a08+ ions are linked to the O atoms of the acid groups and both K+ ions have a coordination number of six (KO6) and have edge-sharing bipyramidal geometries. The K+ ions are bridged by atoms O1 and O3, forming chains propagating along the b-axis direction . The K\u22efO bond lengths vary between 2.6682\u2005(12) and 2.8099\u2005(14)\u2005\u00c5 (Table\u00a03\u2212) bond lengths is much less significant of the organic anion are similar, and similar to that in 4L1_BH , while the conformation of the second (involving atom S2) differs significantly through white to blue . The Hirshfeld surfaces (HS) of 4L1_AH, and 4L1_BH mapped over dnorm are given in Fig.\u00a013a and b concern the O\u2014H\u22efO hydrogen bonds in the crystal structures of both compounds.The Hirshfeld surfaces are colour-mapped with the normalized contact distance, 4L1_AH, and 4L1_BH are shown in Fig.\u00a014The percentage contributions of inter-atomic contacts to the HS for both compounds are compared in Table\u00a094L1_AH, and 4L1_BH, respectively. These are followed by C\u22efH/H\u22efH contacts at, respectively, 4.5 and 4.9%. The N\u22efH/H\u22efN contacts contribute, respectively, 3.0 and 2.5%.The third most important contribution to the HS is from the S\u22efH/H\u22efS contacts at 13.4 and 16.1%, for 4L1_AH, and 4L1_BH, are given in Fig.\u00a015Etot) are also included there; see Tan et al. , the dispersion forces (Edis) and the total energy diagrams (Etot), are shown in Fig.\u00a016et al., 2017et al., 2019\u22121 within a radius of 6\u2005\u00c5 of a central reference mol\u00adecule. It can be seen that for both polymorphs the major contribution to the inter\u00admolecular inter\u00adactions is from electrostatic potential forces (Eele), reflecting the presence of the classical O\u2014H\u22efO hydrogen bonds.A comparison of the energy frameworks calculated for et al., 2016viz. 2,3,5,6-pyrazine\u00adtetra\u00adcarb\u00adoxy\u00adlic acid (pztca) and 2,3,5,6-tetra\u00adkis\u00ad(4-carb\u00adoxy\u00adphen\u00adyl)pyrazine (pztba). Ligand pztba has been shown to be extremely successful in forming metal\u2013organic frameworks potassium] . The asymmetric unit consists of half a mono-deprotonated ligand mol\u00adecule located about an inversion center, and half a potassium ion. The carb\u00adoxy H atom is disordered by symmetry, similar to the situation in the structure of 3L1KH. Here the K\u22efO bond lengths vary from 2.7951\u2005(11) to 2.8668\u2005(13)\u2005\u00c5. The K+ cation has a coordination number of 8 (KO8) and a distorted dodeca\u00adhedral geometry as in 3L1KH .Potassium salts of carb\u00adoxy\u00adlic acids are relatively common. A search for potassium salts of purely organic carb\u00adoxy\u00adlic acids and excluding hydrates, yielded over 200 hits. The potassium salt of L1 Fig.\u00a07a and 11pzdca; Fig.\u00a012-3-carb\u00adoxy\u00adpyrazine-2-carboxyl\u00adato)-di\u00adaqua\u00adpotassium], has been reported -hexa\u00adaqua\u00adbis\u00ad(6-carb\u00adoxy\u00adpyridine-2-carboxyl\u00adato-O)dipotassium potassium] , with the K\u22efO bonds lengths varying from 2.8197\u2005(14) to 3.0449\u2005(15)\u2005\u00c5. The K+ ion has a coordination number of seven (KO6N) and has an edge-sharing penta\u00adgonal anti\u00adprism geometry, forming chains . This structure can be compared to that of 2H2L1K where the two independent K+ ions, each with a coordination number of six (KO6), have edge-sharing bipyramidal geometries, also forming chains .The structure of MUMPIW is that of a potassium-organic framework Fig.\u00a017b, with s Fig.\u00a017b. This ns Fig.\u00a07b and 12et al., 1994The synthesis and crystal structure of the reagent tetra-2,3,5,6-bromo\u00admethyl-pyrazine (TBr) have been reported (Ferigo 4L1):Synthesis of 3,3\u2032,3\u2032\u2032,3\u2032\u2032\u2032-{tetra\u00adkis\u00ad(sulfanedi\u00adyl)}tetra\u00adpropionic acid that X-ray diffraction analysis indicated to be triclinic polymorph 4L1_AH.Mercaptopropionic acid was dissolved in 50\u2005ml THF. A minimum amount of water (a few ml) was added to dissolve 1.4166\u2005g of NaOH. The volume of the mixture was increased to 100\u2005ml by adding THF and the reaction was stirred under reflux for 1\u2005h. Then TBr dissolved in 50\u2005ml THF was added dropwise using an addition funnel. The mixture was stirred under reflux for 6\u2005h. After drying under vacuum, the residue was dissolved in 50\u2005ml of deionized water, and HCl puriss. was added dropwise until a clearly acid pH was obtained. This mixture was stirred at room temperature for 1\u20132\u2005h. The yellow precipitate that formed was filtered off and washed with a minimum amount of water and then CHCl4L1H in methanol gave colourless crystals of rather poor quality. However, X-ray diffraction analysis indicated that a second triclinic .1H NMR , \u03b4(ppm): 4.03 , 2.78 = 7.0, H3), 2.62 = 7.0, H4).13C NMR , \u03b4(ppm): 174.54 , 150.12 , 34.29 , 33.64 , 26.65 .20H28N2O8S4, Mw = 552.71\u2005g\u2005mol\u22121: Calculated: C 43.46, H 5.11, N 5.07%. Found: C 43.40, H 5.17, N 4.87%.Elemental Analysis for Cm/z: 591.04 [M + K]+; 575.06 [M + Na]+; 553.08 [M + H]+; 471.07; 449.09.ESI\u2013MS, \u22121) \u03bd: 2926(s), 2666(m), 2590(s), 1693(s), 1429(s), 1406(s), 1340(m), 1270(s), 1200(s), 1163(m), 1134(s), 1107(m), 1055(w), 918(s), 658(m), 489(m).IR :Synthesis of poly[sulfan\u00adyl]meth\u00adyl}pyrazin-2-yl)meth\u00adyl]sulfan\u00adyl}propano\u00adate)potas\u00adsium] (KH3)2 and 4L1H were mixed together in 20\u2005ml of a 1\u2005M potassium acetate buffer. The mixture was left at 323\u2005K under stirring and nitro\u00adgen conditions for 1\u2005h. The mixture was then filtered and left to evaporate in air for six weeks. Colourless plate-like crystals were obtained, which were shown to be a potassium\u2013organic framework.Hg(NO\u22121) \u03bd: 3422(m), 2922(m), 1713(m), 1580(s), 1399(s), 1247(m), 1190(m), 1152(m), 1114(m), 811(m), 787(m).IR :Synthesis of poly[sulf\u00adan\u00adyl]meth\u00adyl}pyrazine-2,5-di\u00adyl)bis\u00ad(methyl\u00adene)]bis\u00ad(sulfanedi\u00adyl)}dipropionato)dipotassium] (K3)2 and 4L1H were mixed together in 20\u2005ml of a 1M potassium acetate buffer. The mixture was left at 323\u2005K under stirring and nitro\u00adgen for 1\u2005h. The mixture was then filtered and left to evaporate in air for 6 weeks. Colourless plate-like crystals were obtained, which proved to be a dipotassium-organic framework.Zn(NO\u22121) \u03bd: 3401(m), 1579(s), 1401(s), 1303(m).IR . They were therefore included in calculated positions assuming the formation of carb\u00adoxy\u00adlic acid dimers; O\u2014H = 0.82\u2005\u00c5 and refined as riding with Uiso(H) = 1.5Ueq(O).For + salt of pyrazine tetra\u00adcarb\u00adoxy\u00adlic acid = 1.2Ueq(C).For all four compounds, the C-bound H atoms were included in calculated positions and treated as riding on their parent C atom with C\u2014H = 0.97\u2005\u00c5 and 4L1_AH and 4L1_BH, the alert _diffrn_reflns_point_group_measured_fraction_full value below minimum (0.95) was given. For 4L1_AH it involves 131 random reflections out of a total of 2180, viz. 6.0%, while for 4L1_BH it involves 158 random reflections out of a total of 2184, viz. 7.2%.For 4L1_AH, 4L1_BH and 2H2L1K the multiplicity of reflections was 2 or less and so an empirical absorption correction was applied.For 10.1107/S2056989021003479/pk2656sup1.cifCrystal structure: contains datablock(s) H4L1A, H4L1B, KH3L1, K2H2L1, Global. DOI: 10.1107/S2056989021003479/pk2656H4L1Asup2.hklStructure factors: contains datablock(s) H4L1A. DOI: 10.1107/S2056989021003479/pk2656H4L1Bsup3.hklStructure factors: contains datablock(s) H4L1B. DOI: 10.1107/S2056989021003479/pk2656KH3L1sup4.hklStructure factors: contains datablock(s) KH3L1. DOI: 10.1107/S2056989021003479/pk2656K2H2L1sup5.hklStructure factors: contains datablock(s) K2H2L1. DOI: Click here for additional data file.10.1107/S2056989021003479/pk2656H4L1Asup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021003479/pk2656H4L1Bsup7.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021003479/pk2656KH3L1sup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021003479/pk2656K2H2L1sup9.cmlSupporting information file. DOI: 2074770, 2074769, 2074768, 2074767CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two mol\u00adecules A and B are involved in inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds.The title compound, C 9H7NO2S crystallizes with two independent mol\u00adecules (A and B) in the asymmetric unit with Z = 8. Both mol\u00adecules are almost planar with a dihedral angle between the isoxazole and thio\u00adphen rings of 3.67\u2005(2)\u00b0 in mol\u00adecule A and 10.00\u2005(1) \u00b0 in mol\u00adecule B. The packing of mol\u00adecules A and B is of an ABAB\u22ef type along the b-axis direction, the configuration about the C=C bond is Z. In the crystal, the presence of C\u2014H\u22efO, C\u2014H\u22ef N and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances of 3.701\u2005(2) and 3.766\u2005(2)\u2005\u00c5] link the mol\u00adecules into a three-dimensional architecture. An analysis of Hirshfeld surfaces shows the importance of C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds in the packing mechanism of the crystalline structure.The title compound, C In the present study, we report on the synthesis, mol\u00adecular and crystal structure together with a Hirshfeld surface analysis of the title isoxazole derivative.Isoxazolones show some inter\u00adesting biological properties. They are inhibitors of the factorization of tumor necrosis alpha (TNF-\u03b1) (Laughlin A and B) in the asymmetric unit. The mol\u00adecular structure adopts a Z-configuration about the C=C [1.354\u2005(3)\u2005\u00c5 in mol\u00adecule A and 1.357\u2005(3)\u2005\u00c5 in mol\u00adecule B] double bonds. The mol\u00adecular structure of the title compound is shown in Fig.\u00a01. It crysA) and C11\u2014C12\u2014C13 (mol\u00adecule B) differing by 0.8\u2005(2)\u00b0. Also, a slight difference of 0.3\u2005(2)\u00b0 is observed between the angles C2\u2014C5\u2014C6 and C11\u2014C14\u2014C15. In mol\u00adecule A, the angle between the normal of the mol\u00adecular plane (O2A/N1A/C1A\u2013C3A) and the normal of the (S1A/C6A\u2013C9A) plane is 3.67\u2005(2)\u00b0. An important difference is observed in mol\u00adecule B, where the angle between the normal of the mol\u00adecular plane (O3B/N2B/C10B\u2013C12B) and the normal of the (S2B/C15B\u2013C18B) plane is 10.00\u2005(1)\u00b0. In the mol\u00adecular skeleton, the angle between the mean planes of the mol\u00adecules A and B is 4.09\u2005(1)\u00b0. Each of the two methyl groups, C4 and C13, has a C\u2014H bond lying in the mean plane of the mol\u00adecular skeleton, and they are oriented toward the thio\u00adphene group.The bond lengths in the two mol\u00adecules are practically equal, while there are slight differences in bond angles; with for example C2\u2014C3\u2014C4 \u2005\u00c5 and Cg3\u22efCg4ii = 3.766\u2005(2)\u2005\u00c5 where Cg1, Cg2, Cg3 and Cg4 are the centroids of the O2A/N1A/C1A\u2013C3A, S2B/C15B\u2013C18B, S1A/C6A\u2013C9A and O3B/N2B/C10B\u2013C12B rings, respectively; symmetry codes: (i) \u2212x, y, z; (ii) \u2212x, y, z]. The two mol\u00adecules A and B are involved in inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds . The H\u22efH contacts, accounting for about 35.4% of the Hirshfeld surface represent the largest contribution and are seen in the fingerprint plot as a pair of shorts pikes at de + di = 2.2\u2005\u00c5; comparing this to van der Waals radius, we find the difference between them is about 1\u2005\u00c5, which means it is a very powerful inter\u00adaction. H\u22efO/O\u22efH contacts make a contribution of 28.7%, with a distinctive peak in the fingerprint plot at de + di = 2.4\u2005\u00c5; the van der Waals radius sum for this inter\u00adaction is about 2.7\u2005\u00c5.The Hirshfeld surface analysis . Although the H\u22efN /N\u22efH inter\u00adactions have a notable contribution of 12% to the Hirshfeld surface , their inter\u00adatomic distances (de + di = 2.4\u2005\u00c5) are less than their van der Waals radius (2.7\u2005\u00c5), which means that it is a very strong inter\u00adaction in this structure. The presence of \u03c0\u2013\u03c0 stacking reflects the presence of C\u22efC contacts , which account for 7.9% of the Hirshfeld surface with de + di = 3.4\u2005\u00c5; the van der Waals radius is 3.4\u2005\u00c5, so we can confirm the presence of \u03c0\u2013\u03c0 stacking. Two further views of the Hirshfeld surface are shown in Fig.\u00a05The pair of short peaks at ce Fig.\u00a04e, theirts Fig.\u00a04f, whichet al., 2016Z)-4-(thio\u00adphen-2-yl\u00admethyl\u00adidene)isoxazol-5(4H)-one unit gave five hits: 4-(2-hydroxybenzyl\u00adidene)-3-methyl\u00adisoxazol-5(4H)-one -one -4-benzyl\u00adidene-3-methyl\u00adisoxazol-5(4H)-one -one -4-(4-hy\u00addroxy\u00adbenzyl\u00adidene)-3-methyl\u00adisoxazol-5(4H)-one , 10.00\u2005(1), 0.86\u2005(9), 7.02\u2005(8), 2.65\u2005(16), 4.55\u2005(15), 6.50\u2005(1), 7.98\u2005(8) and 3.18\u2005(8)\u00b0, respectively.The asymmetric unit of the title compound contains two crystallographically independent mol\u00adecules, as found for ERIXIN and WOYPIL while in AJESAK, MBYIOZ01 and VIDSAF, there is only one mol\u00adecule per asymmetric unit. The configuration about the C=C bond is via C\u2014H\u22efO hydrogen bonds, forming ABAB chains along the [10A and B is of an ABAB\u22ef type along the [100] direction. In our compound, the cohesion of the crystal is ensured by inter\u00adactions of the type C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 [inter\u00adcentroid distances of 3.701\u2005(2) and 3.766\u2005(2)\u2005\u00c5 compared with 3.811\u2005(2) and 3.889\u2005(2)\u2005\u00c5 in ERIXIN and 3.767\u2005(2) and 3.867\u2005(2)\u2005\u00c5 in WOYPIL].In the crystal of WOYPIL, the individual mol\u00adecules are linked 5H4OS, 1\u2005mmol), hydroxyl\u00adamine hydro\u00adchloride , ethyl aceto\u00adacetate and K2CO3 (5\u2005mol%) were mixed in a 25\u2005mL flask equipped with a magnetic stirrer. The mixture was refluxed in 5\u2005mL of water for 3h (followed by TLC). When the reaction was judged to be finished, the mixture was gradually poured into ice-cold water. Stirring was maintained for a few minutes and the obtained solid was filtered and purified by crystallization from ethanol (yield 72%).Thio\u00adphene-2-carbaldehyde (CUiso(H) = 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details for the title compound are summarized in Table\u00a0210.1107/S2056989021002632/zn2002sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021002632/zn2002Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021002632/zn2002Isup3.cmlSupporting information file. DOI: 2069004CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Broussonetia kazinoki, on vascular endothelial growth factor\u2010A (VEGF\u2010A)\u2013stimulated endothelial cell responses in vitro and microvessel sprouting ex vivo. Treatment with broussonin A or B suppressed VEGF\u2010A\u2010stimulated endothelial cell proliferation by regulating the expression of cell cycle\u2013related proteins and the phosphorylation status of retinoblastoma protein. In addition, treatment with broussonin A or B abrogated VEGF\u2010A\u2010stimulated angiogenic responses including endothelial cell migration, invasion, tube formation and microvessel formation from rat aortic rings. These anti\u2010angiogenic activities of broussonin A and B were mediated through inactivation of VEGF\u2010A\u2010stimulated downstream signalling pathways, localization of vascular endothelial\u2010cadherin at cell\u2010cell contacts, and down\u2010regulation of integrin \u03b21 and integrin\u2010liked kinase. Furthermore, treatment with broussonin A or B inhibited proliferation and invasion of non\u2013small cell lung cancer and ovarian cancer cells. Taken together, our findings suggest the pharmacological potential of broussonin A and B in the regulation of angiogenesis, cancer cell growth and progression.In the present study, we demonstrate the regulatory effects and mechanism of broussonin A and B, diphenylpropane derivatives isolated from Vascular endothelial growth factor (VEGF) has long been appreciated as a key therapeutic target for the treatment of diseases associated with pathological angiogenesis.These complex dynamics of VEGF\u2010mediated signalling networks may partially explain the limitation of anti\u2010VEGF/VEGFR therapy in the clinic. Therefore, in\u2010depth investigations of integrative signalling pathways within the tumour microenvironment may provide crucial therapeutic targets and strategies for improving clinical outcomes.Broussonetia kazinoki (B. kazinoki) Siebold (Moraceae), a deciduous shrub distributed in Eastern Asia including Korea, China and Japan, has been traditionally used as a folk medicine for the treatment of amblyopia, inflammation and oedema as well as a raw material for making high\u2010quality paper in Korea. The extract and bioactive components isolated from B. kazinoki have been reported to possess various pharmacological properties including myogenic, anti\u2010allergic, anti\u2010inflammatory, anti\u2010diabetic and anti\u2010tumour activities.B. kazinoki or marmesin, a furanocoumarin component isolated from B. kazinoki, negatively modulates VEGF\u2010A\u2010induced angiogenic responses by inactivation of VEGF\u2010A/VEGFR\u20102\u2010mediated signalling network.B. kazinoki, B. papyrifera and Anemarrhena asphodeloides (A. asphodeloides), has been reported to possess antiviral, anti\u2010inflammatory, anti\u2010adipogenic and oestrogenic properties.B. kazinoki and A. asphodeloides has been known to exert anti\u2010adipogenic and neurotrophic activities.B. kazinoki on endothelial cell and cancer cell responses.Broussonin A (2\u2010[3\u2010(4\u2010hydroxyphenyl)propyl]\u20105\u2010methoxyphenol), a diphenylpropane derivative isolated from several plants including 22.1\u00ae BulletKit media and used between passages 4 and 6 for all experiments, according to the manufacturer's instructions (Lonza). Human non\u2013small cell lung cancer (A549 and H1299) and ovarian cancer (SKOV\u20103) cells from the American Type Culture Collection were cultured in 10% foetal bovine serum\u2010Dulbecco's modified Eagle's medium .Human umbilical vein endothelial cells (HUVECs) from Lonza were grown in EGM\u201022.2B. kazinoki. High\u2010performance liquid chromatography (HPLC) analysis was performed on an Agilent 1200\u00a0series using Kromasil\u00ae C18 column with a stepwise gradient elution of methanol\u20100.05% trifluoroacetic acid in water (20%\u2013100% methanol) at a flow rate of 1\u00a0mL/min. The purity of broussonin A and B by HPLC analysis was >98%. 1H\u2010 and 13C\u2010nuclear magnetic resonance (NMR) spectra of broussonin A and B were recorded on a Bruker Ascend 700\u00a0MHz NMR spectrometer . Broussonin A: 1H\u2010NMR 7.00 , 6.92 , \u03b4 6.69 , 6.36 , 6.34 , 3.72 , 2.53 , 1.81 ; 13C\u2010NMR \u03b4 158.9 (C\u20104\u2019), 155.6 (C\u20102\u2019), 154.8 (C\u20104'\u2019), 133.5 (C\u20101'\u2019), 129.7 (C\u20106\u2019), 128.9 , 121.0 (C\u20101\u2019), 114.6 , 104.0 (C\u20105\u2019), 100.9 (C\u20103\u2019), 54.1 (OCH3), 34.5 (C\u20103), 32.1 (C\u20102), 28.9 (C\u20101); ESI(electrospray ionization)\u2010MS (mass spectrometry) (positive mode) m/z 259 [M + H]+. Broussonin B: 1H\u2010NMR 6.98 , 6.88 , \u03b4 6.69 , 6.39 , 6.30 , 3.74 , 2.50 , 1.77 ; 13C\u2010NMR \u03b4 158.9 (C\u20102\u2019), 156.9 (C\u20104\u2019), 155.4 (C\u20104'\u2019), 134.1 (C\u20101'\u2019), 130.4 (C\u20106\u2019), 129.5 , 122.0 (C\u20101\u2019), 115.2 , 106.7 (C\u20105\u2019), 98.9 (C\u20103\u2019), 54.8 (OCH3), 35.1 (C\u20103), 32.8 (C\u20102), 29.5 (C\u20101); ESI\u2010MS (positive mode) m/z 259 [M + H]+. The structures of broussonin A and B are presented in Figure\u00a0Broussonin A and B were isolated in an ethyl acetate fraction partitioned from the ethanolic extract of 2.3S6K (T421/S424), anti\u2010phospho\u2010Akt (S473), anti\u2010phospho\u2010ERK (T202/Y204), anti\u2010phospho\u2010p38MAPK (T180/Y182), anti\u2010phospho\u2010pRb (S780) and anti\u2010phospho\u2010pRb (S807/S811) ; anti\u2010phospho\u2010tyrosine ; anti\u2010VEGFR\u20102, anti\u2010vascular endothelial (VE)\u2010cadherin, anti\u2010integrin \u03b21, anti\u2010ILK, anti\u2010p70S6K, anti\u2010Akt, anti\u2010ERK, anti\u2010p38MAPK, anti\u2010Cdk2, anti\u2010Cdk4, anti\u2010cyclin D, anti\u2010cyclin E, anti\u2010actin antibodies and mouse and rabbit IgG\u2010horseradish peroxidase conjugates ; Alexa Fluor 488\u2013conjugated goat anti\u2010mouse IgG .The following agents were obtained from commercial sources: vascular endothelial growth factor\u2010A 165 ; anti\u2010phospho\u2010VEGFR\u20102 (Y951) ; anti\u2010phospho\u2010p702.45 cells/well, SPL Life Sciences, Gyeonggi\u2010do, Republic of Korea) were serum\u2010starved for 14\u00a0h in endothelial cell basal medium\u20102 and pretreated with broussonin A or B (0.1\u201310\u00a0\u00b5M) for 30\u00a0min, followed by VEGF\u2010A (10\u00a0ng/mL) stimulation for 24\u00a0h. In some experiments, human non\u2013small cell lung cancer (A549 and H1299) and ovarian cancer (SKOV\u20103) cells, plated on 6\u2010well plates (5\u00a0\u00d7\u00a0104 cells/well), were serum\u2010starved for 24\u00a0h in basal DMEM and pretreated with broussonin A or B (10\u00a0\u00b5M) for 30\u00a0min, followed by 10% FBS stimulation for 24\u00a0h. Cell proliferation and viability were determined as described previously.HUVECs, plated on 6\u2010well plates .Quiescent HUVECs were pretreated with broussonin A or B (10\u00a0\u00b5M) for 30\u00a0min, followed by VEGF\u2010A (10\u00a0ng/mL) stimulation for 24\u00a0h. Cells were fixed with ice\u2010cold 70% ethanol, stained with Muse\u2122 cell cycle reagent, and then analysed by a Muse2.6TM Total RNA Purification kit . RNA purity and concentration were determined using a NanoDropTM 2000\u00a0spectrophotometer . One microgram of RNA was used as template for RT\u2010mediated PCR using 1st Strand cDNA Synthesis kit . Primer sets for integrin \u03b21 were forward 5\u2019\u2010GAAGGGTTGCCCTCCAGA\u20103\u2019 and reverse 5\u2019\u2010GCTTGAGCTTCTCTGCTGTT\u20103\u2019; and the primer sets for glyceraldehydes\u20103\u2010phosphate dehydrogenase (GAPDH) were forward 5\u2019\u2010GAAGGTGAAGGTCGGAGTC\u20103\u2019 and reverse 5\u2019\u2010GAAGATGGTGATGGGATTTC\u20103\u2019.Total RNA was isolated using PureHelix2.7Quiescent HUVECs were pretreated with broussonin A or B (10\u00a0\u00b5M) for 30\u00a0min, followed by VEGF\u2010A (10\u00a0ng/mL) stimulation for the indicated time points. Cells were lysed by incubation in 50\u00a0mM Tris\u2010HCl (pH 7.4), 1% Triton X\u2010100, 150\u00a0mM NaCl, 0.5\u00a0\u03bcg/mL leupeptin, 1\u00a0\u03bcg/mL pepstatin A, 10\u00a0\u03bcg/mL aprotinin, 100\u00a0\u03bcg/mL 4\u2010(2\u2010aminoethyl)benzenesulfonyl fluoride, 1\u00a0mM EDTA, 1\u00a0mM sodium orthovanadate, 25\u00a0mM sodium fluoride, 80\u00a0mM \u03b2\u2010glycerophosphate and 10% glycerol for 30\u00a0min at 4\u00b0C. Cell lysates were subjected to immunoprecipitation and Western blot as previously described.2.8A single wound was created in the centre of confluent HUVEC monolayer by a sterile pipette tip. After serum starvation for 2\u00a0h, cells were pretreated with broussonin A or B (0.1\u201310\u00a0\u00b5M) for 30\u00a0min, followed by VEGF\u2010A (10\u00a0ng/mL) stimulation for 16\u00a0h. Following fixation with methanol, cells were stained with 0.04% Giemsa solution . The migration of cells across a wound field gap was quantified as previously described.2.9\u00ae (BD Biosciences)\u2010coated transwell inserts were serum\u2010starved for 2\u00a0h and pretreated with broussonin A or B (1\u201310\u00a0\u03bcM) for 30\u00a0min, followed by VEGF\u2010A (10\u00a0ng/mL) or 10% FBS stimulation for 16\u00a0h. After fixation with methanol, invasive cells were stained with 0.04% Giemsa solution and quantified from six different fields using x200 objective magnification.Transwell invasion assay was performed as previously described.2.10Quiescent HUVECs, plated on gelatin\u2010coated coverslips in 12\u2010well plates, were pretreated with broussonin A or B (10\u00a0\u00b5M) for 30\u00a0min, followed by VEGF\u2010A (10\u00a0ng/mL) stimulation for 30\u00a0min. Briefly, cells were fixed with 3.7% paraformaldehyde, permeabilized with 0.1% Triton X\u2010100, blocked with 5% BSA\u2010PBS, and incubated with anti\u2010VE\u2010cadherin antibody. Images were observed using a Carl Zeiss Microscope (Axio Imager.M2) and AxioVision Rel. 4.8\u00a0software .2.114 cells/mL) were plated on Matrigel\u00ae\u2010coated 24\u2010well plates and pretreated with broussonin A or B (1\u201310\u00a0\u00b5M) for 30\u00a0min, followed by VEGF\u2010A (10\u00a0ng/mL) for 6\u00a0h. Formation of capillary\u2010like structures was examined using an Olympus CKX41 inverted microscope (CAchN 10/0.25php objective) and ToupTek Toupview software .After serum starvation for 2\u00a0h, cells (4\u00a0\u00d7\u00a0102.12\u00ae were pretreated with broussonin A or B (10\u00a0\u00b5M) for 30\u00a0min, followed by VEGF\u2010A (500\u00a0ng/mL) for 3\u00a0days and then incubated with fresh broussonin A or B plus VEGF\u2010A every other day, and photographed on the 7th day using x40 objective magnification.Eight\u2010 to nine\u2010week\u2010old male Sprague Dawley rats (250\u00a0\u00b1\u00a010\u00a0g) were purchased from RaonBio Inc. . The animal experiments were conducted in accordance with the institutional guidelines. The experimental procedures were approved by the Institutional Animal Care and Use Committee at Dankook University . Thoracic aortic ring segments embedded in Matrigel2.13t test and was based on at least three different experiments. The results were considered to be statistically significant when p\u00a0<\u00a0.05.Statistical analysis was performed using Student's 33.11 cell cycle arrest, which is well correlated with suppression of cell proliferation \u2010cadherin from \u03b2\u2010catenin and plakoglobin and modulates angiogenic responses such as endothelial permeability, invasion, proliferation and tube formation.3.4S6K), Akt, extracellular signal\u2010regulated kinase (ERK) and p38\u00a0mitogen\u2010activated protein kinase (p38MAPK).S6K, Akt, ERK and p38MAPK, as compared with untreated controls , a key kinase in integrin downstream signalling pathways, which are closely associated with angiogenesis and tumour progression Figure\u00a0.39, 43.5Based on inhibitory effects of broussonin A and B on angiogenic responses, we next examined the ability of broussonin A and B to regulate proliferation and invasion in NSCLC p53 wild\u2010type A549 and p53\u2010deficient H1299 cells as well as ovarian cancer p53\u2010deficient SKOV\u20103 cells Figure\u00a0. Treatme4S6K and p38MAPK, redistribution of VE\u2010cadherin to cell\u2010cell contacts, and down\u2010regulation of integrin \u03b21 and ILK. In addition, both broussonin A and B exert anti\u2010proliferative and anti\u2010invasive activities against NSCLC and ovarian cancer cells. Our findings demonstrate the pharmacological potential of broussonin A and B in the regulation of pathological angiogenic responses associated with cancer growth and progression.In the current study, we report that both broussonin A and B negatively regulate VEGF\u2010A\u2010induced in vitro endothelial cell responses including proliferation, migration, invasion and capillary\u2010like structure formation as well as ex vivo angiogenesis. The mechanism of these anti\u2010angiogenic effects involves inactivation of VEGF\u2010A/VEGFR\u20102 downstream signalling pathways such as ERK, Akt, p70Integrins, transmembrane receptors that facilitate cell\u2010extracellular matrix and cell\u2010cell interactions, mediate a wide range of cellular responses including adhesion, migration, proliferation, invasion and angiogenesis associated with tumour growth and progression.Cell adhesion, migration and invasion are tightly controlled by the changes in the expression of adhesion molecules such as integrins and cadherins and/or activity of matrix metalloproteinases.In addition to anti\u2010angiogenic activity, both broussonin A and B have anti\u2010proliferative and anti\u2010invasive activities against NSCLC and ovarian cancer cells concomitant with suppression of integrin \u03b21 expression, independently of p53\u00a0levels. In conclusion, our results provide significant insights into the regulatory roles and therapeutic efficacy of broussonin A and B in angiogenesis and cancer progression and warrant preclinical evaluation and development as a promising therapeutic agent for the treatment of a wide range of angiogenesis\u2010related diseases including cancer.The authors declare that there are no conflicts of interest.Jae Hyeon Kim: Conceptualization ; Investigation (lead); Methodology (lead); Validation ; Visualization ; Writing \u2013 original draft ; Writing \u2013 review & editing . Sunho Kim: Data curation ; Formal analysis ; Investigation . Surim Han: Data curation ; Formal analysis ; Investigation . Eun\u2010Kyung Ahn: Conceptualization (supporting); Data curation ; Investigation (supporting). Young\u2010Rak Cho: Conceptualization (supporting); Data curation ; Investigation (supporting). Wonsik Jeong: Data curation ; Investigation (supporting). Sung Joon Kim: Data curation (supporting); Investigation (supporting). Gyu\u2010Un Bae: Conceptualization (supporting); Resources (supporting). Joa Sub Oh: Conceptualization (supporting); Resources (supporting). Dong\u2010Wan Seo: Conceptualization (lead); Data curation ; Funding acquisition (lead); Resources (lead); Supervision (lead); Writing \u2013 original draft (lead); Writing \u2013 review & editing (lead).Supplementary MaterialClick here for additional data file."} +{"text": "Three intra\u00admol\u00adecular hydrogen bonds are observed in the title compound. In the crystal, mol\u00adecules are connected by C\u2014H\u22efCl and N\u2014H\u22efO hydrogen bonds. 18H15ClN3O+\u00b7Cl\u2212\u00b72H2O, three intra\u00admol\u00adecular hydrogen bonds are observed, N\u2014H\u22efO, O\u2014H\u22efCl and O\u2014H\u22efO. In the crystal, mol\u00adecules are connected by C\u2014H\u22efCl and N\u2014H\u22efO hydrogen bonds. Strong C\u2014H\u22efCl, N\u2014H\u22efO, O\u2014H\u22efCl and O\u2014H\u22efO hydrogen-bonding inter\u00adactions are implied by the Hirshfeld surface analysis, which indicate that H\u22efH contacts make the largest contribution to the overall crystal packing at 33.0%.In the title compound, C For example, their biological activity and anti\u00admicrobial properties have been researched extensively (Neumann Database survey section). Three intra\u00admol\u00adecular hydrogen bonds are observed, N2\u2014H2C\u22efO2, O2\u2014H2A\u22efCl2 and O2\u2014H2B\u22efO3 between the pyridazine rings [centroid\u2013centroid distance = 3.4902\u2005(12)\u2005\u00c5], and also between the pyridine and benzene rings [3.7293\u2005(13) and 3.8488\u2005(13)\u2005\u00c5], forming sheets.The water mol\u00adecules and chloride anions are located in channels between the organic cations and are connected by O\u2014H\u22efO and O\u2014H\u22efCl hydrogen bonds Table\u00a01 into chant Fig.\u00a02b betweeConQuest version 2021.3.0; Groom et al., 2016H)pyridazinone -one -one on the Hirshfeld surface imply strong hydrogen-bonding inter\u00adactions of types C\u2014H\u22efCl, N\u2014H\u22efO, O\u2014H\u22efCl and O\u2014H\u22efO. In the shape-index map , the \u03c0\u2013\u03c0 inter\u00adactions are indicated by the red and blue triangles. Fig.\u00a04c and Fig.\u00a04d show di and de surfaces and Fig.\u00a04e and 4f the curvedness and fragment path surfaces. Fig.\u00a05a shows the overall two-dimensional fingerprint plot. The fingerprint plot delineated into H\u22efH contacts has a point with the tip at de + di = 2.05\u2005\u00c5. The pair of wings in the fingerprint plot defined into H\u22efC/C\u22efH contacts (19.3 percent contribution to the HS), Fig.5c, has a pair of thin edges at de + di \u223c2.99\u2005\u00c5 while the pair of wings for the H\u22efCl/Cl\u22efH contacts are seen as two spikes with the points at de + di = 2.97\u2005\u00c5 and de + di = 2.41\u2005\u00c5. The fingerprint plot for H\u22efO/O\u22efH contacts has two spikes with the tips at de + di = 2.11\u2005\u00c5 and de + di = 1.83\u2005\u00c5. As seen in Fig.\u00a05f the C\u22efC contacts (7.4%) have an arrow-shaped distribution of points with tips at de + di = 3.37\u2005\u00c5. The contributions of the N\u22efH/H\u22efN contacts to the Hirshfeld surface (5.8%) are less important . Fig.\u00a06To investigate the effect of the mol\u00adecular inter\u00adactions on the crystal packing, the Hirshfeld surface Fig.\u00a03 and fingap Fig.\u00a04b, the \u03c0nt Fig.\u00a05g. Fig.\u00a0et al., 2019E)-6-(4-chloro\u00adstyr\u00adyl)-4,5-di\u00adhydro\u00adpyridazin-3(2H)-one and nicotinaldehyde in 30\u2005ml of ethanol, sodium ethano\u00adate was added. The mixture was refluxed for 3\u2005h. The reaction mixture was cooled, diluted with cold water and acidified with concentrated hydro\u00adchloric acid. The precipitate was filtered, washed with water, dried and recrystallized from ethanol. White single crystals were obtained by slow evaporation at room temperature, yield 86%; m.p. 453\u2005K; FT\u2013IR (KBr): \u03bd 3322 (NH), 1651 (C=O), 1584\u2005cm\u22121 (C=N); 1H NMR \u03b4 13.20 , 8.98 , 8.83 , 8.57 , 8.05 8.02 , 7.65 , 7.45 , 7.36 , 7.08 , 4.09 ppm ; 13C NMR \u03b4 160.43, 145.98, 143.89, 141.87, 140.05, 139.25, 137.97, 134.90, 132.84,130.85, 128.82, 128.62, 128.54, 126.80, 125.08, 32.33 ppm. ESI-MS: m/z = 324.08 [M+H]+.The title compound was synthesized according to a previously published procedure (Daoui Uiso(H) = 1.2 Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022003346/jq2014sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022003346/jq2014Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022003346/jq2014Isup3.cmlSupporting information file. DOI: 2161716CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the mol\u00adecules are linked by C\u2014H\u22efN inter\u00adactions along the a-axis direction, forming a C(6) chain. The mol\u00adecules are further connected by C\u2014Cl\u22ef\u03c0 inter\u00adactions and face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions, forming ribbons along the a-axis direction. Hirshfeld surface analysis indicates that the greatest contributions to the crystal packing are from Cl\u22efH/H\u22efCl (35.1%), H\u22efH (10.6%), C\u22efC (9.7%), Cl\u22efCl (9.4%) and C\u22efH/H\u22efC (9.2%) inter\u00adactions.In the title compound, C The secondary important H\u22efH and C\u22efC inter\u00adactions contribute 10.6% and 9.7% , respectively, to the Hirshfeld surface. The remaining contributions for the title compound are from Cl\u22efCl, C\u22efH/H\u22efC, Cl\u22efF/F\u22efCl, Cl\u22efC/C\u22efCl, F\u22efH/H\u22efF, N\u22efH/H\u22efN, N\u22efN and F\u22efC/ C\u22efF contacts, which are less than 9.7% and have a negligible effect on the packing. The percentage contributions of all inter\u00adactions are listed in Table\u00a03The overall two-dimensional fingerprint plot and those delineated into Cl\u22efH/H\u22efCl, H\u22efH, C\u22efC, Cl\u22efCl and C\u22efH/H\u22efC contacts in the title mol\u00adecule are illustrated in Fig.\u00a076% Fig.\u00a07c and 9.7% Fig.\u00a07d, respeet al., 2016E)-1--2-phenyl\u00addiazene unit resulted in 28 hits. Nine compounds are closely related to the title compound, viz. LEQXOX and 60.31\u2005(14)\u00b0, respectively. In I, C\u2014H\u22efN and short Cl\u22efCl contacts are observed and in II, C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and short C\u2014Cl\u22efO contacts occur. In III, the benzene rings form a dihedral angle of 63.29\u2005(8)\u00b0 and the mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds into zigzag chains running along the c-axis direction. The crystal packing also features C\u2014Cl\u22ef\u03c0, C\u2014F\u22ef\u03c0 and N\u2014O\u22ef\u03c0 inter\u00adactions. In IV, the benzene rings make a dihedral angle of 56.13\u2005(13)\u00b0. Mol\u00adecules are stacked in columns along the a-axis direction via weak C\u2014H\u22efCl hydrogen bonds and face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions. The crystal packing is further consolidated by short Cl\u22efCl contacts. In V and VI, the aromatic rings form dihedral angles of 60.9\u2005(2) and 64.1\u2005(2)\u00b0, respectively. In the crystals, mol\u00adecules are linked through weak X\u22efCl contacts (X = Cl for V and Br for VI), C\u2014H\u22efCl and C\u2014Cl\u22ef\u03c0 inter\u00adactions into sheets parallel to the ab plane. Additional van der Waals inter\u00adactions consolidate the three-dimensional packing. In VII, the dihedral angle between the two aromatic rings is 64.12\u2005(14)\u00b0. The crystal structure is stabilized by a short C\u2014H\u22efCl contact, C\u2014Cl\u22ef\u03c0 and van der Waals inter\u00adactions. In VIII, the benzene rings subtend a dihedral angle of 77.07\u2005(10)\u00b0. In the crystal, mol\u00adecules are associated into inversion dimers via short Cl\u22efCl contacts [3.3763\u2005(9)\u2005\u00c5]. In IX, the asymmetric unit comprises two similar mol\u00adecules, in which the dihedral angles between the two aromatic rings are 70.1\u2005(3) and 73.2\u2005(2)\u00b0. The crystal structure features short C\u2014H\u22efCl and C\u2014H\u22efO contacts and C\u2014H\u22ef\u03c0 and van der Waals inter\u00adactions.In the crystals of et al., 2018E)-1--2-(4-fluoro\u00adbenzyl\u00adidene)hydrazine , tetra\u00admethyl\u00adethylenedi\u00adamine (TMEDA) , CuCl and CCl4 . After 1\u20133\u2005h , the reaction mixture was poured into \u223c0.01 M solution of HCl , and extracted with di\u00adchloro\u00admethane (3 \u00d7 20\u2005mL). The combined organic phase was washed with water (3 \u00d7 50\u2005mL) and brine (30\u2005mL), dried over anhydrous Na2SO4 and concentrated using a vacuum rotary evaporator. The residue was purified by column chromatography on silica gel using appropriate mixtures of hexane and di\u00adchloro\u00admethane (3/1\u20131/1). Crystals suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution. Colourless solid (44%); m.p. 345\u2005K. Analysis calculated for C14H7Cl4FN2 (M = 364.02): C 46.19, H 1.94, N 7.70; found: C 46.11, H 1.98, N 7.67%. 1H NMR \u03b4 7.31\u20137.83 . 13C NMR \u03b4 114.89, 115.12, 115.41, 115.74, 115.97, 118.33, 127.73, 128.08, 128.67, 129.17, 130.48, 132.04, 132.15 and 136.83. ESI\u2013MS: m/z: 365.11 [M + H]+.The title dye was synthesized according to the reported method = 1.2Ueq(C). Five outliers Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989021010756/vm2255sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021010756/vm2255Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021010756/vm2255Isup3.cmlSupporting information file. DOI: 2116300CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Each Tb atom is coordinated by two cyclo\u00adpenta\u00addienyl ligands in an \u03b7 5H5)2(C14H12NO2)], was synthesized from tris\u00ad(cyclo\u00adpenta\u00addien\u00adyl)(tetra\u00adhydro\u00adfuran)\u00adterbium and 2-{[(2-meth\u00adoxy\u00adphen\u00adyl)imino]\u00admeth\u00adyl}phenol. Each Tb atom is coordinated by two cyclo\u00adpenta\u00addienyl ligands in an \u03b75-coordination mode and by one N and two O atoms of the organic ligand in a tridentate \u03ba3O,N,O\u2032-mode.The air- and moisture-sensitive title compound, [Tb(C Such a combination of ligands provides an extremely broad structural diversity for cyclo\u00adpenta\u00addienyl derivatives (2-{[(2-meth\u00adoxy\u00adphen\u00adyl)imino]\u00admeth\u00adyl}phenolato)terbium, which is a product of the partial protonation of the tris(cyclo\u00adpenta\u00addien\u00adyl)terbium complex with 2-{[(2-meth\u00adoxy\u00adphen\u00adyl)imino]\u00admeth\u00adyl}phenol . Assuming that each cyclo\u00adpenta\u00addienyl ligand donates three electron pairs, the terbium atom may be considered to be ennea-coordinated. Both cyclo\u00adpenta\u00addienyl ligands are nearly symmetrically \u03b75-coordinated to the Tb3+ cation. Thus, the Cp(centroid)\u2014Tb distances [2.4207\u2005(11)\u2005\u00c5 for the C1\u2013C5 Cp ring and 2.4062\u2005(10)\u2005\u00c5 for the C6\u2013C10 Cp ring] are almost equal to the Cp(plane)\u2014Tb distances [2.4196\u2005(11)\u2005\u00c5 for C1\u2013C5 Cp ring and 2.4054\u2005(10) for C6\u2013C10 Cp ring], and the CCp\u2014Tb bond lengths are similar within each ring (Table\u00a01Cp\u2014Tb distance to the C1\u2013C5 ring is longer by 0.011\u2005\u00c5 than to the second Cp ligand. Such a slight asymmetry is caused by the presence of the tridentate asymmetric 2-{[(2-meth\u00adoxy\u00adphen\u00adyl)imino]\u00admeth\u00adyl}phenolate (L\u2212) ligand. Atoms of the ligand are situated in two planes formed by the following sets of atoms: O1, C11\u2013C16, N1, C24 (r.m.s. deviation = 0.0167\u2005\u00c5) and O2, C17\u2013C23, N1 (r.m.s. deviation = 0.0333\u2005\u00c5). The dihedral angle between these planes of 44.58\u2005(5)\u00b0 indicates a perceptible loss of conjugation between two parts of the ligand due to the tridentate \u03ba3N,O,O\u2032-coordination mode. The bond redistribution within the ligand (see table in the supporting information) and the Tb\u2014O and Tb\u2014N bond distances 2Ln(O2NC14H12)] were previously synthesized in low yields or protonated (LH) derivatives have been poorly studied, whereas the structures of complexes bearing their closest analogs \u2013 doubly charged 2-{[(2-oxidophen\u00adyl)imino]\u00admeth\u00adyl}phenolate and its various derivatives \u2013 have been studied moderately. This is, likely, due to the higher stability of the latter complexes, which is presumably caused, in short, by a higher degree of the optimization of electrostatic inter\u00adactions 3(thf) was obtained according to a literature procedure 3(thf) in 15\u2005ml of THF. The reaction mixture was stirred for 24\u2005h. The solution was concentrated under vacuum to a volume of ca 8\u201310\u2005ml, and hexane (10\u2005ml) was carefully layered on top of the resulting solution to initiate crystallization. Crystals obtained after several days were dried under dynamic vacuum for 1\u2005h, yielding 0.315\u2005g . The terbium content was determined by direct complexometric titration with the disodium salt of EDTA, using xylenol orange indicator imino]\u00admeth\u00adyl}phenol in 5\u2005ml of THF was added slowly to a solution of Tb = 1.5Ueq(C) for methyl group and Uiso(H) = 1.2Ueq(C) for others.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021013025/yk2162sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021013025/yk2162Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021013025/yk2162Isup3.cdxSupporting information file. DOI: 2127087CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Starting from this ligand monomeric [M(4\u2010BzhH2BoxCH)] (M=Na (7), K (18)) and dimeric [{M(4\u2010BzhH2BoxCH)}2] (M=K (28), Rb (9), Cs (10)) alkali metal complexes were synthesised by deprotonation. Abstraction of the potassium ion of 8 by reaction with 18\u2010crown\u20106 resulted in the solvent separated ion pair [{(THF)2K@(18\u2010crown\u20106)}{bis(4\u2010benzhydryl\u2010benzoxazol\u20102\u2010yl)methanide}] (11), including the energetically favoured monoanionic \u2010(4\u2010BzhH2BoxCH) ligand. Further reaction of 4\u2212BzhH2BoxCH2 with three equivalents KH and two equivalents 18\u2010crown\u20106 yielded polymeric [{(THF)2K@(18\u2010crown\u20106)}{K@(18\u2010crown\u20106)K(4\u2010BzhBoxCH)}]n (n\u2192\u221e) (12) containing a trianionic ligand. The neutral ligand and herein reported alkali complexes were characterised by single X\u2010ray analyses identifying the latter as a promising precursor for low\u2010valent main group complexes.A novel sterically demanding bis(4\u2010benzhydryl\u2010benzoxazol\u20102\u2010yl)methane ligand 4\u2212BzhH2BoxCH2)Bis(4\u2010benzhydryl\u2010benzoxazol\u20102\u2010yl)methane ( provides a versatile platform for monomeric ligands in [M(4\u2010BzhH2BoxCH)] and dimeric [{M(4\u2010BzhH2BoxCH)}2] alkali metal complexes. Even more remarkable the reaction of 4\u2212BzhH2BoxCH2 with three equivalents KH and two equivalents 18\u2010crown\u20106 yields polymeric [{(THF)2K@(18\u2010crown\u20106)}{K@(18\u2010crown\u20106)K(4\u2010BzhH2BoxCH)}]n (n\u2192\u221e), containing a trianionic ligand. While the monoanions only provide N,N\u2010chelation like NacNac the latter trianion potentially provides C,N,N,C\u2010chelation and should be another promising ligand platform. N,N\u2019\u2010ligands like \u03b2\u2010diketiminatesDippNacNac . These findings initiated research into other ligand platforms, which mimic the coordination abilities of the ubiquitous NacNac ligand, in particular the \u03ba2\u2010N,N\u2010coordination of the two imine nitrogen atoms to a metal ion to form six\u2010membered metallaheterocycles. Starting from the bis(2\u2010pyridyl)methane ligand and its isoelectronically N,3N2M, M=cationic metal fragment) is maintained. In recent years the focus has been on symmetrical bis(benzoxazole\u20102\u2010yl)methanides I to IV in Scheme\u20053\u2010linker unit ethylbisimidate dihydrochloride derived from malonic acid.n\u2010butyllithium or potassium hydride.X2Al(\u2212I)]X=Cl, I). Despite of the steric demand provided by iPr\u2010 or tBu\u2010residues at C4\u2010position close to the coordination pocket in III or IV, mimicking the bulkiness provided by NacNac ligands, low\u2010oxidation or low\u2010valent Group 2 or 13 elements could so far not be synthesised. Therefore, we decided to introduce bulkier benzhydryl groups at the ligand scaffold inspired by compounds containing 2,6\u2010dibenzhydrylphenyl residues e.\u2009g. [:MH(L)]M=Ge or Sn, (L)=\u2010N(Ar)(SiiPr3), Ar=4\u2010iPr\u2010C6H2Bzh2) or [M(ArN)2CN=CtBu2]Ar=4\u2010tBu\u2010C6H2Bzh2). In the following paragraph, the six\u2010step synthesis of bis(4\u2010benzhydryl\u2010benzoxazol\u20102\u2010yl)methane and its behaviour in alkali metal coordination are discussed.Over the last decades, monoanionic bidentate 6, 4\u2010BzhH2BoxCH2), 2\u2010amino\u20103\u2010benzhydrylphenol was first synthesised according to the synthetic protocols of Quaranta et\u2005al.1\u20135 see Supporting Information). Thereafter, two equivalents of 2\u2010amino\u20103\u2010benzhydrylphenol and one equivalent of ethyl cyanoacetimidate hydrochloride C3\u2010linker unit were reacted by cyclocondensation reaction.) (7), starting material 6 and neat sodium metal were vigorously stirred until all metal was consumed (5 d). After all volatiles had been removed under reduced pressure, 7 was isolated as a white powder in excellent yields (98\u2009%). Crystals suitable for single crystal XRD experiments were grown from a saturated toluene solution at \u221228\u2009\u00b0C. Donor\u2010free 7 crystallises in the monoclinic space group P21/n with one molecule in the asymmetric unit ] (7) exhibits a distorted \u03ba2\u2010N,N\u2019\u2010coordination of the sodium ion by the imine ring nitrogen atoms. A negligible dislocation of the cation from the C3N2\u2010plane of 0.117(2) \u00c5 and a minor butterfly folding angle of 4.82(6)\u00b0 between the two benzoxazol\u20102\u2010yl moieties is observed (Table\u20057 (Na1\u2212N1 2.3392(16) \u00c5; Na1\u2212N2 2.3456(15) \u00c5) are similar to the Na1\u2212N1 2.358(6) \u00c5 bond of the six\u2010membered sodium \u03b2\u2010diketiminate complex [(THF)2NaL]3)C(Ph)}2CH) but are slightly shorter than neutral five\u2010membered TMEDA based [[{Na{N(SiMe3)(Dipp)}(TMEDA)}2] (Na1\u2212N2 2.4726(17) \u00c5; Na1\u2212N3 2.461(2) \u00c5)iPrNC(NAr)(HNiPr)}(THF)]2 (Na1\u2212N1 2.453(1) \u00c5, Na1\u2212N2 2.558(2) \u00c5).+ ion \u03c0\u2010arene, interactions are formed to one phenyl group of the benzhydryl groups each.Ar distances ranging from 2.8305(18) (Cortho) to 3.0414(18) \u00c5 (Cipso) indicate a dihaptic arene coordination.1H NMR spectra of 6 and deprotonated species 7 show a significant deshielding in toluene and simultaneous downfield shift of the methanide bridge from \u03b4(\u2212H2C\u2212)=3.85\u2005ppm to \u03b4(\u2212HC\u2212)=5.38\u2005ppm whereas benzylic protons experience a significant upfield shift from \u03b4(\u2212HCPh2)=6.41\u2005ppm to 5.27\u2005ppm. Furthermore, arene protons (Ph and NCOC6H3) are slightly upfield shifted in sodium complex 7 compared to ligand 6. Elemental analysis and mass spectrometry confirmed the synthesis of bulk [Na(4\u2010BzhH2BoxCH)] (7).6 with potassium hydride (Scheme\u20054\u2010BzhH2BoxCH)]n with n=1 (18) and 2 (28), (YLD: 89\u2009%). Colourless crystals were grown from a saturated toluene solution at \u221230\u2009\u00b0C after 1 d. Single crystal XRD experiments revealed a monomeric as well as a dimeric species in the solid state. Monomeric [(toluene)K(4\u2010BzhH2BoxCH)] crystallises in the triclinic space group P1\u203ewith one molecule and half a toluene molecule in the asymmetric unit \u00c5 is present. Due to the larger radius of potassium, longer K\u2212N (K1\u2212N1 2.7768(18) \u00c5; K1\u2212N2 2.6982(18) \u00c5) and a less acute N1\u2212K1\u2212N2 of 70.73(5)\u00b0 are observed. This is accompanied by an increasing butterfly folding angle to 5.54(8)\u00b0. Furthermore, potassium phenyl carbon distances suggest that the cation is \u03b72\u2010(K1\u2212C30 3.467(2) \u00c5, K1\u2212C31 3.224(2) \u00c5) and \u03b73\u2010coordinated (K1\u2212C23 3.314(2) \u00c5, K1\u2212C27 3.503(2) \u00c5, K1\u2212C28 3.236(2) \u00c5) by one phenyl residue of each benzhydryl group (A search in the CCDC version 5.41 (Aug. 2020)\u03b76\u2010coordinated ones provided a range of short contacts between K and Cortho of 2.77 to 3.76\u2005\u00c5). (K\u2212H 2.64(4) to 3.37(3) \u00c5, K\u2212H\u2010C 117.8 to 120.7\u00b0)Ar)2] [M{N(SiMe3)2}3] .9 and 10 also display interactions of metal ions and methanide bridges, but due to the increased residual electron density for the heavier homologues the determination of the freely refined hydrogen atom positions is not that reliable. Rising effective ionic radii,The second species derived from 0) YLD: 7\u2009% comple0) YLD: 7\u2009% comple0) YLD: 7\u2009% compleZ,Z)\u2010 and \u2010(4\u2010BzhH2BoxCH) (28: 2.775(2) to 3.015(2) \u00c5; 9: 2.905(5) to 3.063(4) \u00c5; 10: 3.033(3) to 3.285(3) \u00c5), more acute average N\u2212M\u2212N angles as well as a growing dislocation of the metal from the C3N2 plane (28: 1.877(4) to 2.282(3) \u00c5; 9: 2.036(7) to 2.303(7) \u00c5; 10: 2.302(5) to 2.430(5) \u00c5) as well as from the \u2212HC2N unit (28: 0.85(5) to 2.18(3) \u00c5; 9: 0.699(13) to 2.196(9) \u00c5; 10: 1.16(8) to 2.60(5) \u00c5). Interestingly, the detected butterfly folding angles in both isomers \u2010(4\u2010BzhH2BoxCH)\u224823\u00b0; \u2010(4\u2010BzhH2BoxCH)\u224816\u00b0) are negligibly affected while coordinating the alkali metal ions. Additionally, all alkali metal complexes 28\u201310 were studied by 1H and 13C{1H} NMR spectroscopy in [D8]THF. Recorded 1H NMR spectra show a distinctive pattern of chemical shifts arising from the C2v symmetry of monoanionic bis(benzoxazol\u20102\u2010yl)methanide scaffold and the benzylic bound protons, whereas phenylic protons of the benzhydryl groups could not be clearly assigned due to their peak overlap. Deprotonation of 6 results in an accumulation of electron density in the benzene periphery of both benzoxazol\u20102\u2010yl moieties that again entails a general shielding of corresponding protons and an upfield shift. Concerning the 1H NMR shifts for the C\u2212H bridging position, a minute but continuous decline along K (4.66\u2005ppm)>Rb (4.65\u2005ppm)>Cs (4.64\u2005ppm) complexes is noticed. The opposite is found for para positioned protons and benzylic protons. These protons exhibit a small but noticeable downfield shift =6.33\u2005ppm to 6.43\u2005ppm; \u03b4=6.03\u2005ppm to 6.17\u2005ppm) which is in tune with the negative charge being restored in the phenylic benzhydryl groups yielding a growing metal arene interaction. Remaining benzoxazol\u20102\u2010yl protons in ortho\u2010 and meta\u2010position shifts do not seem to be influenced by varying alkali cations. Although 1H NMR spectra of 8\u201310 ([D8]THF) and sodium complex 7 ([D8]toluene) were measured in different solvents, benzylic protons display a remarkable change of the average chemical shifts (\u25b5\u03b4=0.82\u2005ppm). The 87Rb NMR spectrum of 9 from THF solution shows two singlets. Most probably [{Rb(4\u2010BzhH2BoxCH)}2] rearranges to give a THF\u2010solvated rubidium cation [Rb(THF)n] (\u03b4=\u22121.69\u2005ppm) and a bis(4\u2010benzhydryl\u2010benzoxazol\u20102\u2010yl)methanide} based anion [Rb(4\u2010BzhH2BoxCH)2]\u2212 (\u03b4=\u2212254.69\u2005ppm). The 133Cs NMR spectrum of 10 displays only one signal at \u03b4=\u221231.12\u2005ppm. Additionally, elemental analysis and mass spectrometry confirmed the synthesis of heavier alkali complexes 8\u201310.These features Table\u2005 cause riZ,Z)\u2010 and \u2010isomers of complex 8\u201310 and based on former gained knowledge of sterically less demanding bis(benzoxazol\u20102\u2010yl)methanide potassium complexes+ [4\u2010BzhH2BoxCH]\u2212 (11) was isolated as a reddish solid in excellent yields (95\u2009%). Crystals suitable for single XRD experiments were grown by vapour diffusion of pentane to a saturated THF solution of 11 at ambient temperature after 3 d. The colourless crystals in the triclinic space group P1\u203econtain one bis(4\u2010benzhydryl\u2010benzoxazol\u20102\u2010yl)methanide anion and two halve (THF)2K(18\u2010crown\u20106)} cations in the asymmetric unit K(18\u2010crown\u20106)] . (11) was confirmed by mass spectrometry and elemental analysis.On the basis of reported 2K(18\u2010crown\u20106)}{K2(4\u2010BzhBoxCH)}]n (n\u2192\u221e) 12 as very sensitive dark purple crystals by storing THF solutions at \u221230\u2009\u00b0C. The crystal selection and mounting was challenging due to the dark colour and high sensitivity of compound 12. Thus, a moderate single crystal XRD data set could be collected after many attempts. Furthermore, the synthesis of 12 was improved by using ligand 6 and the exact stoichiometric quantities (4\u2010BzhH2BoxCH2:KH:18\u2010crown\u20106=1\u2009:\u20093\u2009:\u20092) . Obtained THF solution was cooled to \u221230\u2009\u00b0C for crystallisation. Complex 12 crystallises in the monoclinic space group P2/n with half a formula unit in the asymmetric unit \u00c5, C5\u2212K2 3.245(8) \u00c5) to the aryl moieties of two adjacent \u2010trianions and also coordinated by an equatorial crown ether molecule. The \u03b72\u2010coordination provides the link in the polymeric structure. Finally, a third potassium ion in the coordination pocket is surrounded by two nitrogen atoms in \u03ba2\u2010fashion (N\u2212K1 2.686(3) \u00c5) and a phenyl ring of each benzhydryl moiety in asymmetrically \u03b76\u2010fashion (CPh\u2212K1: 2.930(3) \u00c5 to 3.291(3) \u00c5), respectively. Determined nitrogen potassium distances and the perfect planarity of the ligand scaffold with the potassium ion in the plane of the trianion are in good agreement with the unsolvated \u03ba2\u2010N,N\u2010[K(DippNacNac)]ipso\u2010atoms are basically shortened (1.426(4) to 1.478(4) \u00c5) compared to HCbzh\u2212Cipso of complexes 6\u201311 (1.508(8) \u00c5 to 1.538(6) \u00c5) because of the deprotonation of the two benzylic positions (Cbzh) (benzhydryl group). The two anionic C9\u2010atoms are essentially in plane with the bound Cipso\u2010atoms similar to previously studied potassium trityl complexes [KCPh3(THF)(PMDTA)],3(PMDTA)]3(L)]n .2\u2010hybridised carbon atoms (for details see Supporting Information), while the steric constraints prevent the phenyl rings from being coplanar leading to a propeller\u2010like arrangement. Within this arrangement, angles \u03c6 of the phenyl groups with respect to C2\u2212C10\u2212C16 plane (C3\u2010plane) are \u03c6=32.38(17)\u00b0 (C10 to C15), and 49.68(10)\u00b0 (C1 to C6), whereas the smallest angle and shortest bond (C9\u2212C16 1.426(4) \u00c5) involves the phenyl system to which the cation is coordinated. These parameters correspond to an increased \u03c0\u2010electron\u2010delocalisation that is correlated with the overlap of the C3\u2010plane (C2\u2212C10\u2212C16) and coordinating phenyl ring (C16 to C21) which is dependent on cos(\u03c6)2 function\u03c6=17.7\u00b0,\u224890\u2009%). Further direct analyses of 12 were challenging due to its low solubility in most solvents . To find further evidence for the synthesis of trifold deprotonated anion (4\u2010BzhBoxCH), suspensions of the dark red precipitate (12) were once again protonated (excess\u224820\u2005eq. H2O or D2O) in small scale (NMR experiment). These 1H and 2D NMR experiments (for details see Supporting Information), as well as mass spectrometry , confirmed the previous synthesis of 12.Since Brown or Buncel and Menon ascertained that triphenylmethane is deprotonated in THF by potassium hydride when DMFwn\u20106)}{K2\u2010BzhBoxCH6 (4\u2212BzhH2BoxCH2) comprising benzhydryl groups at both C4\u2010positions (ortho imine positions) in spatial proximity to the coordination pocket was presented. To get a better knowledge of its properties and find a possible precursor complex for subsequent salt metathesis reactions, 6 was deprotonated with alkali metal bases. Obtained products were analysed by NMR spectroscopy, mass spectrometry as well as single crystal XRD experiments. Crystals grown from toluene solutions unveiled monomeric [M(4\u2010BzhH2BoxCH)] (M=Na (7), K (18)) and dimeric [{M(4\u2010BzhH2BoxCH)}2] (M=K (28), Rb (9), Cs (10)) species in solid state. Latter alkali metal complexes display distorted \u03ba2\u2010N,N\u2010coordinated \u2010 and an \u2010(4\u2010BzhH2BoxCH) configurational isomers, which display various polyhaptic metal arene interactions. Furthermore, potassium ion sequestration of 8 by 18\u2010crown\u20106 resulted in [{(THF)2K(18\u2010crown\u20106)}(4\u2010BzhH2BoxCH)] (11) a solvent separated ion pair containing energetically favoured monoanionic \u2010(4\u2010BzhH2BoxCH) ligand. Additionally, reaction of 4\u2010BzhH2BoxCH2 with three equivalents KH and two equivalents 18\u2010crown\u20106 yielded polymeric [{(THF)2K@(18\u2010crown\u20106)}{K@(18\u2010crown\u20106)K(4\u2010BzhBoxCH)}]n (n\u2192\u221e) (12) featuring a remarkable trianionic ligand. The single crystal X\u2010ray diffraction experiment of 12 revealed one of the potassium ions to be \u03b72\u2010coordinated by the benzoxazol\u20102\u2010yl scaffold of two adjacent \u2010ligands plus a crown ether molecule. A second potassium ion is surrounded by two nitrogen atoms in \u03ba2\u2010fashion as well as one phenyl ring of each benzhydryl moiety in a symmetrically \u03b76\u2010fashion, respectively. Future work with regard to the mono\u2010 (4\u2010BzhH2BoxCH) and trianionic (4\u2010BzhBoxCH) ligand will focus on the coordination of other metal, i.\u2009e. those from Group 13 or lanthanides. The benzhydryl groups attached to the benzoxazol\u20102\u2010yl scaffold seem to exhibit similar qualities to the triphenylmethane, which is able to stabilise carbanionic, radical or carbonium ionic species owing to the extensive \u03c0\u2010delocalisation. With this in mind, future research might focus on the synthesis of 4\u2010BzhH2BoxCH2 based radical and carbonium ion compounds.Within this work, the six\u2010step synthesis and characterisation of a novel bulky bis(benzoxazol\u20102\u2010yl)methane ligand d6, thf\u2010 d8, toluene\u2010 d8).[45] Deuterated benzene and toluene were dried over K (65\u2009\u00b0C), THF was pre\u2010dried with LiAlH4 and stored over activated molecular sieve (3\u2005\u00c5) and stored in an argon dry box. Elemental analyses were performed on a Vario EL3 at the Mikroanalytische Labor, Institut f\u00fcr Anorganische Chemie, University of G\u00f6ttingen. LIFDI\u2010MS spectra were measured on a Jeol AccuTOF spectrometer and ESI (HR\u2010MS) measurements were performed on Bruker maXis spectrometer. All pKa measurements in acetonitrile were carried out as in reference [13]. Shock\u2010cooled crystals were selected from a Schlenk flask under argon atmosphere using the X\u2010TEMP2 device.6, 18, 9, 10, 11) or Ag\u2010 I\u03bcS microfocus source.6 a 3\u03bb correction was applied.2 using the full\u2010matrix least\u2010squares methods of SHELXL6),2031908 (for 7), 2031909 (for 81), 2031910 (for 82), 2031911 (for 9), 2031912 (for 10), 2031913 (for 11), and 2031914 (for 12)Deposition Numbers 2031907 and 2D NMR spectroscopic data were recorded on a Bruker Ascend 500\u2005MHz, 400\u2005MHz, and Avance 300\u2005MHz spectrometer and referenced to the deuterated solvent should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "The structures of the six hydrogen-bonded 1:1 compounds of 4-methyl\u00adquinoline with 2-chloro-4-nitro\u00adbenzoic acid, 2-chloro-5-nitro\u00adbenzoic acid, 2-chloro-6-nitro\u00adbenzoic acid, 3-chloro-2-nitro\u00adbenzoic acid, 4-chloro-2-nitro\u00adbenzoic acid and 5-chloro-2-nitro\u00adbenzoic acid have been determined at 185\u2013190\u2005K. In each crystal, the acid and base mol\u00adecules are linked by a short hydrogen bond between a carb\u00adoxy/carboxyl\u00adate O atom and an N atom of the base. 10H9N) with chloro- and nitro-substituted benzoic acids (C7H4ClNO4), namely, 4-methyl\u00adquinolinium 2-chloro-4-nitro\u00adbenzoate, C10H10N+\u00b7C7H3ClNO4\u2212, (I), 4-methyl\u00adquinoline\u20132-chloro-5-nitro\u00adbenzoic acid (1/1), C10H9N\u00b7C7H4ClNO4, (II), 4-methyl\u00adquinolinium 2-chloro-6-nitro\u00adbenzoate, C10H9.63N0.63+\u00b7C7H3.37ClNO40.63\u2212, (III), 4-methyl\u00adquinolinium 3-chloro-2-nitro\u00adbenzoate, C10H9.54N0.54+\u00b7C7H3.46ClNO40.54\u2212, (IV), 4-methyl\u00adquinolinium 4-chloro-2-nitro\u00adbenzoate, C10H10N+\u00b7C7H3ClNO4\u2212, (V), and 4-methyl\u00adquinolinium 5-chloro-2-nitro\u00adbenzoate, C10H10N+\u00b7C7H3ClNO4\u2212, have been determined at 185\u2013190\u2005K. In each compound, the acid and base mol\u00adecules are linked by a short hydrogen bond between a carb\u00adoxy (or carboxyl\u00adate) O atom and an N atom of the base. The O\u22efN distances are 2.5652\u2005(14), 2.556\u2005(3), 2.5485\u2005(13), 2.5364\u2005(13), 2.5568\u2005(13) and 2.5252\u2005(11)\u2005\u00c5, respectively, for compounds (I)\u2013(VI). In the hydrogen-bonded acid\u2013base units of (III) and (IV), the H atoms are each disordered over two positions with O site:N site occupancies of 0.37\u2005(3):0.63\u2005(3) and 0.46\u2005(3):0.54\u2005(4), respectively, for (III) and (IV). The H atoms in the hydrogen-bonded units of (I), (V) and (VI) are located at the N-atom site, while the H atom in (II) is located at the O-atom site. In all the crystals of (I)\u2013(VI), \u03c0\u2013\u03c0 stacking inter\u00adactions between the quinoline ring systems and C\u2014H\u22efO hydrogen bonds are observed. Similar layer structures are constructed in (IV)\u2013(VI) through these inter\u00adactions together with \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of the adjacent acid mol\u00adecules. A short Cl\u22efCl contact and an N\u2014O\u22ef\u03c0 inter\u00adaction are present in (I), while a C\u2014H\u22efCl hydrogen bond and a \u03c0\u2013\u03c0 inter\u00adaction between the benzene ring of the acid mol\u00adecule and the quinoline ring system in (II), and a C\u2014H\u22ef\u03c0 inter\u00adaction in (III) are observed. Hirshfeld surfaces for the title compounds mapped over dnorm and shape index were generated to visualize the weak inter\u00admolecular inter\u00adactions.The structures of the six hydrogen-bonded 1:1 compounds of 4-methyl\u00adquinoline (C Ka values of the acids and bases as well as the inter\u00admolecular inter\u00adactions in the crystals. In our ongoing studies of crystal structures for the system of quinoline derivatives\u2013chloro- and nitro-substituted benzoic acids, we have shown that three compounds of quinoline with 3-chloro-2-nitro\u00adbenzoic acid, 4-chloro-2-nitro\u00adbenzoic acid and 5-chloro-2-nitro\u00adbenzoic acid \u2013 pKa(acid)] values of these compounds are in the range 2.93\u20133.38. Although the pKa value of 4-methyl\u00adquinoline is 5.66, which is slight larger than quinoline (pKa = 4.90) and 6-methyl\u00adquinoline (pKa = 5.20), the system of 4-methyl\u00adquinoline\u2013chloro- and nitro-substituted benzoic acids is an attractive candidate for studying short hydrogen bonds and also weak inter\u00admolecular inter\u00adactions. We report here crystal structures of six hydrogen-bonded compounds, namely, 4-methyl\u00adquinolinium 2-chloro-4-nitro\u00adbenzoate, (I)Ka values are 3.62, 3.44, 4.04, 3.84, 3.69 and 3.80, respectively, for (I)\u2013(VI) (Table\u00a01The properties of hydrogen bonds formed between organic acids and organic bases depend on the p) Table\u00a01.The mol\u00adecular structures of compounds (I)\u2013(VI) are shown in Fig.\u00a01abKa value of each compound are also given in Table\u00a01The dihedral angles made by the benzene C1\u2013C6 ring, the carb\u00adoxy/carboxyl\u00adate O1/C7/O2 plane and the nitro O3/N1/O4 plane of the acid, and the quinoline N2/C8\u2013C16 ring system of the base in each hydrogen-bonded acid-base unit of (I)\u2013(VI) are summarized in Table\u00a01In all the compounds of 3-chloro-2-nitro\u00adbenzoic acid and 4-chloro-2-nitro\u00adbenzoic acid, the nitro O3/N1/O4 group is approximately perpendicular to the benzene C1\u2013C6 ring with dihedral angles of 74.4\u2005(3)\u201388.54\u2005(13)\u00b0, while in the 2-chloro-6-nitro\u00adbenzoic acid mol\u00adecule of compound (III)Ka value is observed for each system of quinoline and 6-methyl\u00adquinoline compounds, while for the title compounds (I)\u2013(VI) this correlation is somewhat low.The correlation between the H-atom position in the short hydrogen bond and the \u0394pCg2\u22efCg2, Cg2\u22efCg3 and Cg3\u22efCg3, are 3.4323\u2005(7)\u20133.7751\u2005(8), 3.5878\u2005(7)\u20133.9304\u2005(9) and 3.7719\u2005(8)\u20133.9227\u2005(9)\u2005\u00c5, respectively, where Cg2 and Cg3 are the centroids of the N2/C8\u2013C11/C16 and C11\u2013C16 rings of the quinoline ring system, respectively. The base mol\u00adecules in the crystals of (I)via these \u03c0\u2013\u03c0 inter\u00adactions, while in (III)\u2013(VI) inversion-related base mol\u00adecules are alternately stacked in column-like structures. On the other hand, \u03c0\u2013\u03c0 inter\u00adactions between the inversion-related acid mol\u00adecules are only observed in crystals (IV)\u2013(VI); the centroid-centroid distances, Cg1\u22efCg1, are 3.5702\u2005(7)\u20133.8602\u2005(6)\u2005\u00c5, where Cg1 is the centroid of the C1\u2013C6 ring. Detailed supra\u00admolecular features in the crystals formed through these \u03c0\u2013\u03c0 inter\u00adactions combined with other weak inter\u00admolecular inter\u00adactions are described below.In all the crystals of (I)\u2013(VI), \u03c0\u2013\u03c0 inter\u00adactions between the quinoline ring systems, related by an inversion centre to each other, are observed. The centroid\u2013centroid distances between the quinoline ring systems, namely, via \u03c0\u2013\u03c0 inter\u00adactions between the quinoline ring systems . The dimeric units are further linked via a C\u2014H\u22efO hydrogen bond \u2005\u00c5. Between the layers, an N\u2014O\u22ef\u03c0 inter\u00adaction \u2005\u00c5; symmetry code: (v) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a02] and a C\u2014H\u22efO hydrogen bond on Fig.\u00a02. The ribon Fig.\u00a02 via anotvia C\u2014H\u22efO hydrogen bonds x, y\u00a0+\u00a01, z; (vi) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z]. The acid\u2013base units are linked via these \u03c0\u2013\u03c0 inter\u00adactions, forming a ribbon structure along the b-axis direction on Fig.\u00a04. The tapon Fig.\u00a05.i, C13\u2014H13\u22efO2ii and C14\u2014H14\u22efCg1ii; symmetry codes as in Table\u00a04c-axis direction \u2212x, \u2212y, \u2212z\u00a0+\u00a01; (v) \u2212x\u00a0+\u00a01, \u2212y, \u2212z\u00a0+\u00a01].In the crystal of (III)on Fig.\u00a06. The basms Fig.\u00a07, and thua-axis direction \u2212x\u00a0+\u00a01, \u2212y, z\u00a0+\u00a01; (iv) \u2212x, \u2212y, \u2212z\u00a0+\u00a01]. The ribbons are further linked into a layer parallel to the (011) plane of 3.6685\u2005(8)\u2005\u00c5 . The layers are linked by a C\u2014H\u22efO hydrogen bond on Fig.\u00a08 via C\u2014H\u22efne Fig.\u00a09 via a \u03c0\u2013a-axis direction , 3.7751\u2005(8), 3.7870\u2005(8), 3.9304\u2005(9) and 3.7719\u2005(8)\u2005\u00c5, respectively, for Cg1\u22efCg1vi, Cg2\u22efCg2iii, Cg2\u22efCg3ii, Cg2\u22efCg3iii and Cg3\u22efCg3ii . Between the layers, a C\u2014H\u22efO hydrogen bond is observed n Fig.\u00a010 via a C\u2014C2/c) is different from those of (IV)Pvia \u03c0\u2013\u03c0 inter\u00adactions between the quinoline ring systems and C\u2014H\u22efO hydrogen bonds . The ribbons are connected into a layer parallel to (10via a weak \u03c0\u2013\u03c0 inter\u00adaction between adjacent acid rings with Cg1\u22efCg1iv = 3.8602\u2005(6)\u2005\u00c5 .Uiso(H) = 1.5Ueq(N or O); the refined distances are given in Tables 4Uiso(H) = 1.2 or 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0810.1107/S2056989021010896/hb7991sup1.cifCrystal structure: contains datablock(s) global, I, II, III, IV, V, VI. DOI: 10.1107/S2056989021010896/hb7991Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021010896/hb7991Isup8.cmlSupporting information file. DOI: 10.1107/S2056989021010896/hb7991IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989021010896/hb7991IIsup9.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021010896/hb7991IIIsup10.cmlSupporting information file. DOI: 10.1107/S2056989021010896/hb7991IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989021010896/hb7991IVsup11.cmlSupporting information file. DOI: 10.1107/S2056989021010896/hb7991IVsup5.hklStructure factors: contains datablock(s) IV. DOI: Click here for additional data file.10.1107/S2056989021010896/hb7991Vsup12.cmlSupporting information file. DOI: 10.1107/S2056989021010896/hb7991Vsup6.hklStructure factors: contains datablock(s) V. DOI: Click here for additional data file.10.1107/S2056989021010896/hb7991VIsup13.cmlSupporting information file. DOI: 10.1107/S2056989021010896/hb7991VIsup7.hklStructure factors: contains datablock(s) VI. DOI: 2116680, 2116679, 2116678, 2116677, 2116676, 2116675CCDC references: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Structures of three tetrahalophthalic anhydrides , Br (TBPA), I (TIPA)) were studied by X-ray diffraction, and several types of halogen bonds (HaB) and lone pair\u00b7\u00b7\u00b7\u03c0-hole (lp\u00b7\u00b7\u00b7\u03c0h) contacts were revealed in their structures. HaBs involving the central oxygen atom of anhydride group (further X\u00b7\u00b7\u00b7O(anhydride) were recognized in the structures of TCPA and TBPA. In contrast, for the O(anhydride) atom of TIPA, only interactions with the \u03c0 system (\u03c0-hole) of the anhydride ring (further lp(O)\u00b7\u00b7\u00b7\u03c0h) were observed. Computational studies by a number of theoretical methods demonstrated that the X\u00b7\u00b7\u00b7O(anhydride) contacts in TCPA and TBPA and lp(O)\u00b7\u00b7\u00b7\u03c0h in TIPA are caused by the packing effect. The supramolecular architecture of isostructural TCPA and TBPA was mainly affected by X\u00b7\u00b7\u00b7O(acyl) and X\u00b7\u00b7\u00b7X HaBs, and, for TIPA, the main contribution provided I\u00b7\u00b7\u00b7I HaBs. Charge transfer (CT) complexes are widely applied in different fields including sensors , ferroelIn the context of the discussion of noncovalent interactions, X\u00b7\u00b7\u00b7X ,14,17,18Our experience in halogen bonding investigations ,21,22,23Literature structures of TXPA include one structure for TIPA: BOPDOZ, three for TCPA: TECLPA01-3, and five for TBPA: TBPHAN and TBPHAN01\u20134. TIPA and all TCPA structures were determined in the 1970s\u201380s and have poor quality by modern requirements. Because of this reason, we obtained a series of high-quality crystals of tetrahalophthalic anhydrides (including TBPA) and redetermined their structures (R \u2264 3%) using modern XRD equipment , C\u2013X\u00b7\u00b7\u00b7O(anhydride), and C\u2013X\u00b7\u00b7\u00b7X, where X is halogen atom was carried out for each crystal with the aim to investigate the contribution of all intramolecular interactions in crystal packing (molecular surfaces are depicted in As it follows from Further contacts are considered in the order of their uniqueness and significance of the contribution to the supramolecular structure.RvdW [C\u2013X\u00b7\u00b7\u00b7O(anhydride) interaction between the halogen atom (Cl2/Br2) and the anhydride oxygen atom (O1) is the distinctive feature of crystal packing of TCPA and TBPA. The distances C4\u2013Cl2\u00b7\u00b7\u00b7O1 and C4\u2013Br2\u00b7\u00b7\u00b7O1 (3.0972(14) \u00c5 and 3.1873(19) \u00c5, respectively, RvdW ) \u00b7\u00b7\u00b7\u03c0h bifurcate interaction between anhydride oxygen and two adjacent carbon atoms of a fused furan ring was found. The distances O1\u00b7\u00b7\u00b7C7 and O1\u00b7\u00b7\u00b7C8 were 3.057(4) \u00c5 and 3.127(4) \u00c5, respectively, and the appropriate \u03a3H-carbazole (CCDC code VILFIF) and its N-methylated analogue (CCDC code WEXKEP) as well as the crystal of 2,3-dichloromaleic anhydride (CCDC code LIZCOM). The geometrical parameters of C\u2013X\u00b7\u00b7\u00b7O(anhydride) interactions in these structures are presented in vdW radii, the conducted theoretical DFT calculations short contacts: the co-crystals of pyromellitic dianhydride with 3,6-dibromo-9arene interaction was observed only for TIPA.Besides C\u2013X\u00b7\u00b7\u00b7O(anhydride) contact in TCPA and TBPA, several types of HaB were observed in each crystal structure, which include HaB with acyl oxygen atom C\u2013X\u00b7\u00b7\u00b7O=C and type II halogen\u2013halogen contacts X\u00b7\u00b7\u00b7X. These interactions were presented in structures of all TXPA, whereas I\u00b7\u00b7\u00b7CRvdW . The angles \u2220(C\u2013X\u00b7\u00b7\u00b7O) varied in the range 173.08(7)\u2013177.59(9)\u00b0, satisfying the IUPAC criteria for HaB [In the structures of TXPA each molecule forms a HaB with the acyl oxygen. The X\u00b7\u00b7\u00b7O(acyl) distances equaled 3.0373(16) \u00c5, 3.093(2) \u00c5, and 3.141(3) \u00c5, respectively, which were shorter than the appropriate \u03a3 for HaB . NoncovaRvdW = 3.50 \u00c5); the distance Br1\u00b7\u00b7\u00b7Br2 was 3.5816(4) \u00c5 (vs. \u03a3RvdW was 3.70 \u00c5). The angles \u2220(C3\u2013Cl1\u00b7\u00b7\u00b7Cl2) = 174.58(7)\u00b0 and \u2220(C1\u2013Br1\u00b7\u00b7\u00b7Br2) = 173.89(8)\u00b0 satisfied the criteria of HaB [In the crystals of TCPA and TBPA, there are two types of halogen atoms, and one behaves as an acceptor and one as a donor of X\u00b7\u00b7\u00b7X HaB. The contact Cl1\u00b7\u00b7\u00b7Cl2 is rather elongated (the interatomic distance was 3.4978(7) \u00c5 vs. \u03a3a of HaB . The HaBa of HaB .RvdW (3.96 \u00c5). The angles \u2220(C4\u2013I2\u00b7\u00b7\u00b7I1) = 174.00(8)\u00b0 and \u2220(C5\u2013I3\u00b7\u00b7\u00b7I3) = 157.67(9)\u00b0 are consistent with the IUPAC criteria for HaB [The arrangement of C\u2013I\u00b7\u00b7\u00b7I bonds in TIPA is different from the noncovalent C\u2013X\u00b7\u00b7\u00b7X bonding in the TCPA and TBPA structures described above. Each molecule of TIPA has two HaB donor iodine atoms, which form contacts C4\u2013I2\u00b7\u00b7\u00b7I1 and C5\u2013I3\u00b7\u00b7\u00b7I3. The distances (3.7497(6) \u00c5 and 3.7760(5) \u00c5, respectively) were shorter than \u03a3 for HaB . The I3 RvdW . In the case of TBPA, the distance of Br4\u00b7\u00b7\u00b7C2 was slightly longer than the \u03a3RvdW (3.584(3) \u00c5 vs. 3.55 \u00c5).The from-the-atom lp(X)\u00b7\u00b7\u00b7\u03c0h interaction involving halogen atoms and a furan ring was observed in the TCPA and TBPA crystals . The disRvdW was 3.22 \u00c5. Additionally, another acyl oxygen atom O3 in the TCPA molecule takes part in from-the-atom lp(O)\u00b7\u00b7\u00b7\u03c0h binding O3\u00b7\u00b7\u00b7C8 with the furan ring (the distance was 3.209(2) \u00c5 vs. \u03a3RvdW = 3.22 \u00c5).In the cases of TCPA and TBPA, the acyl oxygen O2 forms from-the-bond lp(O)\u00b7\u00b7\u00b7\u03c0h bonding with C1 and C2 atoms of the fused five-membered ring. The distances O2\u00b7\u00b7\u00b7C1 and O2\u00b7\u00b7\u00b7C2 varied in the range of 2.939(3)\u20132.951(2) \u00c5 and 3.076(4)\u20133.043(3) \u00c5 for TCPA and TBPA, correspondingly, whereas the appropriate \u03a3Contrastingly, in the TIPA crystal lp\u00b7\u00b7\u00b7\u03c0h interactions were realized only at the expense of the O1 atom of the furan ring as discussed above.The strength and nature of interactions were investigated for the most relevant contacts (C\u2013X\u00b7\u00b7\u00b7O (acyl or anhydride) and C\u2013X\u00b7\u00b7\u00b7X, where X is a halogen atom) by a number of theoretical methods: the molecular electrostatic potential (MEP) ,28, the The MEP surface is important to understand the nature of noncovalent interactions regulated by electrostatic effects ,28. On tb) and Laplacian (\u22072\u03c1b) and total energy density (Hb) at the BCP. Low values of \u03c1b (0.008\u20130.010 a.u.), positive values f \u22072\u03c1b (0.024\u20130.040 a.u.), and virtually zero values of Hb are typical for HaBs of weak strength [2)\u03c1b (from \u22120.010 to \u22120.020 a.u.) indicates that these interactions between tetrahalophthalic anhydrides were attractive and that they fall into the van der Waals domain \u03c1b(r) function mapped on the \u03b4ginter isosurface and the 2D plot of the \u03b4ginter descriptor against sign(\u03bb2)\u03c1b(r) are shown in The contacts in question were additionally examined and visualized by the independent gradient model method. The IGMH approach provides a quantitative reference for the characterization of noncovalent interactions, such as the fully noninteracting gradient reference |\u2207\u03c1The charge transfer (CT) is another important factor that may determine HaB. The second-order perturbation energy (E(2)) and the charge transfer value (\u2206occ) can be used to measure the intermolecular interactions . One canIn order to provide further insight into the electron density changes upon the formation of a halogen bond, electron density difference maps (EDD) for bimolecular fragments tetrahalophthalic anhydrides were calculated and the results are illustrated in Red regions represent the accumulation of electron density as a result of the formation of the complex, and blue regions indicate loss of electron density. The EDD plots also show that polarization effects caused by the positive \u03c3-hole developed on the halogen atom tend to shift electron density from donor atoms and hence increase the electron density in the intermolecular space between halogen and oxygen atoms. The polarization of electron density between the halogen and donor atoms clearly implies the formation of the X\u00b7\u00b7\u00b7O/X interaction, which was also illustrated by the results of the NBO analysis .int) into its components , exchange (Eexch), induction (Eind), and dispersion (Edis) terms). Here, the SAPT0 level of theory was used to determine the Eint of the HaB bond in the bimolecular fragments in the X-ray structures changed, which evidenced the critical role of packaging effects. Optimization of the TBPA fragment led to a contact switch from Br\u00b7\u00b7\u00b7O(anhydride) to two Br\u00b7\u00b7\u00b7O(carboxyl). The optimization of TIPAF1 led to a collapse of the structure, which was rearranged to the layered geometry. Thus, this interaction was dominated by the dispersion character over the electrostatic, which possibly led to the formation of a layered structure. Therefore, the O\u00b7\u00b7\u00b7C interactions between TIPA molecules were not sufficiently strong to be preserved in a gas phase.Finally, we investigated the dependence of X/C\u00b7\u00b7\u00b7O(anhydride) interactions on packing effects. To this aim, the full geometry optimization of the bimolecular fragments was perf2Br2, and CH2I2 respectively.Tetrahlophthalic anhydrides are commercially available compounds and were purchased from Merck. Crystals of TCPA, TBPA, and TIPA suitable for X-ray studies were obtained via slow evaporation of solutions of the corresponding anhydrides in DMSO, CHwww.ccdc.cam.ac.uk/data_request/cif (accessed on 22 May 2021).X-ray diffraction studies were performed at 100 K on an Xcalibur Eos diffractometer (for TBPA and TIPA) using Mo-K\u03b1 (\u03bb = 0.71073 nm) radiation and SuperNova diffractometer (for TCPA) using Cu-K\u03b1 (\u03bb = 0.154184 nm) radiation. All structures were solved by direct methods by means of the SHELX program incorporWave function calculations for the QTAIM, IGM, EDD, and NBO analyses were carried out using the crystallographic coordinate at the DFT PBE0 ,61 levelThe QTAIM, IGM, and EDD calculations were carried out using the Multiwfn 3.8 software ,71,72. TIn this work, we studied noncovalent interactions in the crystals of TXPA and recognized hitherto undescribed X\u00b7\u00b7\u00b7O(anhydride) short contacts in the structures of TCPA and TBPA. Based on the analysis of geometrical parameters, these contacts have been classified as HaBs. Contrastingly, in the crystal structure of TIPA, such HaB was not observed and anhydride oxygen was involved in the lp\u00b7\u00b7\u00b7\u03c0h interaction with C atoms of the anhydride system.int = \u22121.8 and \u22122.3 kcal/mol for TCPA and TBPA, respectively) and were apparently caused by packing effects. The structure of the TIPA crystal was predominantly directed by the strong I\u00b7\u00b7\u00b7I HaBs with a lesser role for the C\u2013I\u00b7\u00b7\u00b7O(acyl) interaction , which provide another crystal packing profile and, hence, the absence of X\u00b7\u00b7\u00b7O(anhydride) contact. In addition, the performed calculations established that described X\u00b7\u00b7\u00b7O(anhydride) HaBs have an electrostatic and dispersive nature.Theoretical calculations demonstrated that the greater contribution to the architecture of TCPA and TBPA crystals were made by the C\u2013X\u00b7\u00b7\u00b7O(acyl) and C\u2013X\u00b7\u00b7\u00b7X HaBs, whereas X\u00b7\u00b7\u00b7O(anhydride) HaBs were weaker (EOn the one hand, these findings should be taken into account when using TXPA as CT acceptors because formation of close contacts may influence the CT complex structure. On the other hand, the ability of TXPA to form different types of non-covalent interactions can be useful for crystal engineering purposes and makes these species attractive as building blocks for the assembling of supramolecular constructions."} +{"text": "N-haloimides form two types of short (imide)X\u00b7\u00b7\u00b7N and X\u2013X\u00b7\u00b7\u00b7N halogen bonds. Nucleophilic substitution or ligand-exchange reaction on the peripheral X of X\u2013X\u00b7\u00b7\u00b7N with the chloride of N-chlorosuccinimide lead to Cl\u2013X\u00b7\u00b7\u00b7N halogen-bonded complexes. The 1:1 complexation of HMTA and ICl manifests the shortest I\u00b7\u00b7\u00b7N halogen bond [2.272(5) \u00c5] yet reported for an HMTA acceptor. Two halogen-bonded organic frameworks are prepared using 1:4 molar ratio of HMTA and N-bromosuccinimide, each with a distinct channel shape, one possessing oval and the other square grid. The variations in channel shapes are due to tridentate and tetradentate (imide)Br\u00b7\u00b7\u00b7N coordination modes of HMTA. Density Functional Theory (DFT) studies are performed to gain insights into (imide)X\u00b7\u00b7\u00b7N interaction strengths (\u0394Eint). The calculated \u0394Eint values for (imide)Br\u00b7\u00b7\u00b7N are smaller than the values for (imide)I\u00b7\u00b7\u00b7N . The DFT additivity analysis of (imide)Br\u00b7\u00b7\u00b7N motifs demonstrates Br\u00b7\u00b7\u00b7N interaction strength gradually decreasing from 1:1 to 1:3 HMTA:N-bromosuccinimide complexes. Exceptionally similar charge density values \u03c1(r) for N\u2013I covalent bond and I\u00b7\u00b7\u00b7N non-covalent bond of a (saccharin)N\u2013I\u00b7\u00b7\u00b7N motif signify the covalent character for I\u00b7\u00b7\u00b7N halogen bonding.Hexamethylenetetramine (HMTA) and Non-aromatics, such as 1,4-diazabicylcooctane (DABCO) and hexamethylenetetramine (HMTA), have become a critical design trait in supramolecular chemistry, and have inspired scholars to devise molecular rotors and 2.432(3) \u00c5] are longer than in [DABCO]\u00b7[NBS]2 [2.347(2) and 2.364(2) \u00c5] due to steric and competitive HB interactions in the former structure. (ii) Solvent as the only varying parameter, six different [HMTA]\u00b7[N-iodosuccinimide]4 XBOFs have been characterized using X-ray diffraction analysis find the \u2033ideal\u2033 HMTA:N-haloimide partner to access open-framework XBOF structures, and (iii) evaluate and compare the X\u00b7\u00b7\u00b7N interaction strengths.Our previous contributions in the field of HMTA halogen bonding include (i) the comparison of Br\u00b7\u00b7\u00b7N bond distances in bidentate complexes [HMTA]\u00b7[N-bromosuccinimide]1\u201313 of composition types [HMTA]\u00b7[dihalogen]n and [HMTA]\u00b7[N-haloimide]n motifs, and complexes 2, 4, and 6 comprising hetero-halogen Y\u2013X\u00b7\u00b7\u00b7N motifs. The Br2 source in 1 and I2 in 3 and 5 are consequences of N\u2013X bond cleavage reactions of NBS and NIS, respectively 2N\u2013Br\u00b7\u00b7\u00b7N gradually converts to Br\u2013Br\u00b7\u00b7\u00b7N motif by N\u2013Br bond cleavage reaction followed by the exchange of (CO)2N and Br anions. In the second step, NCS, a chloride anion source, was added to replace the terminal bromide anion to give 2.HMTA and five different N-haloimides, namely N-chlorosuccinimide (NCS), N-bromosuccinimide (NBS), N-iodosuccinimide (NCS), N-bromophthalimide (NBP), and N-iodosaccharin (NISac), were used to prepare 13 halogen-bonded complexes oimide]n . [HMTA]\u00b7 Filler, . The pre7\u201313 were obtained by slow evaporation of the corresponding HMTA and N-haloimide solutions . The 1:4 HMTA:NBS and 1:4 HMTA:NBP molar ratio reactions only gave the corresponding bidentate complexes 7 and 8 as the main products for structural analysis. Single-crystals isolated from different 1:4 HMTA:NBS experiments carried out by using various solvents also revealed the bidentate coordination for HMTA . Using a 1:4 HMTA:NBS molar ratio, complexes 9 and 10 were obtained employing different crystallization techniques. Complex 9, which contains a tridentate Br\u00b7\u00b7\u00b7N HMTA, was obtained using solvent-assisted grinding followed by solution crystallization, while 10, determined to be tetradentate Br\u00b7\u00b7\u00b7N HMTA, was crystallized by using the layering technique. Under the crystallization method of 9, the other HMTA-imide combinations produced crystals of either HMTA or corresponding imide. Even 9 is not reproducible and yields crystals of succinimide and bidentate complex 7. The lack of reproducibility is due to the influence of several uncontrolled factors in the crystallization process, such as N\u2013X bond cleavage reactions and complex hydrogen bonding patterns.Single-crystals of 11. A 1:2 HMTA:NISac molar ratio gives a monodentate complex 12. However, treating HMTA with an excess of NISac (12.5 eq) leads to the formation of unknown quantities of iodine-oriented ions consequently resulting in an iodonium complex [bis(HMTA)I]+13. A related method to prepare [bis(HMTA)I]+2 and [HMTA]\u00b7[Ditfb]2, respectively. The corresponding XB parameters are used for discussions here and their structural data is included in the Supporting Information.Experiments conducted using 1:4 molar ratio of HMTA and NIS in different solvents all exclusively produced crystals of 1\u201313 illustrate four types of coordination modes. All the X\u00b7\u00b7\u00b7N distances are below the sum of the Van der Waals radii of respective halogen and nitrogen (1.55 \u00c5) are shown in HMTA in complexes ) Bondi, . Selecte1 and bidentate in 2 (3-center-4-electron [N\u2013Br\u2013N]+ XBs [2.086(5)\u22122.1862(4) \u00c5] \u22122.1572(2) \u00c5] \u00c5] is remarkably short compared to I\u2013I\u00b7\u00b7\u00b7N distance of 3 [2.405(5) \u00c5] indicating better e-accepting power of ICl. The monodentate Cl\u2013I\u00b7\u00b7\u00b7N distance is shorter than +I\u2013N halogen bonds reported for [bis(HMTA)I]++ XBs [2.198(3)\u22122.349(18) \u00c5]. In the packing structure, 3 associates via HMTAN\u2013I\u2013I\u00b7\u00b7\u00b7NHMTA halogen bonds at distances of 3.519 \u00c5 forming zigzag 1D chains \u00c5], 5 [2.480(4) \u00c5], and [HMTA]\u00b7[I\u2013I]3 [2.593(6) \u00c5] and 2.426(4) \u00c5] are slightly shorter than our previously reported solvent-free bidentate [HMTA]\u00b7[NBS]2 [2.414(3) and 2.432(3) \u00c5] \u00c5, HMTA is bidentate in molecule . The two9 prepared by solvent-assisted manual grinding, contains four crystallographically independent NBS donors of which one NBS does not participate in the X-bonding. Three Br\u00b7\u00b7\u00b7N distances vary from 2.371(8) to 2.411(9) \u00c5 (3/unit cell. The relative channel volume (rcv) \u00c5 . The fouolecules , which a11 contains bidentate HMTA, and the I\u00b7\u00b7\u00b7N distances \u00c5, 2 [2.879(5) and 2.864(4) \u00c5] distances of [DMAP]\u00b7[NIS]. However, both the (imide)N\u2013I [2.26(2) \u00c5] and I\u00b7\u00b7\u00b7N(HMTA) distances [2.29(2) \u00c5] of 12 are remarkably close to [DMAP]\u00b7[NISac] [2.292(2) and 2.218(2) \u00c5] and 2.299(15) \u00c5, and appear within 0.02 \u00c5 of the corresponding distances in the reported structure (Complex 18(2) \u00c5] . Complextructure .S, min, \u221230 kcal/mol), is comparable to values estimated at the O-atoms of NBS and NBP . A VS, max of magnitude +16 kcal/mol is associated with the \u2013CH2- protons adjacent to the sp3 N-atom. In NBS, the VS, max at the bromine \u03c3-hole and the five-member ring-centroid are similar is slightly smaller than in NBS but larger than in NBP. To our surprise, the electron-withdrawing \u2013SO2 group of NISac could only render a positive potential of +7 kcal/mol at the six-membered ring-centroid. The global MEP analysis suggests that the nucleophilic and electrophilic sites of HMTA and NBS/NIS molecules have equal propensity to form Br\u00b7\u00b7\u00b7N, C\u2013H\u00b7\u00b7\u00b7N, and C\u2013H\u00b7\u00b7\u00b7O=C interactions, which is in good agreement with packing forces discussed in the XBOF structures X\u00b7\u00b7\u00b7N motif in a 1:1 complex when donors are successively added to HMTA, are estimated for complexes [HTMA]\u00b7[NBS]int) were estimated by using their corresponding X-ray crystal structure coordinates. The calculated \u0394Eint values range from \u221211.2 to \u221212.5 kcal/mol for Br\u00b7\u00b7\u00b7N motifs. The \u0394Eint values decrease when the HMTA denticity increases, \u221212.5 kcal/mol for 7, \u221212.1 kcal/mol for 9, and \u221211.2 kcal/mol for 10, and the results are in good agreement with the additivity analysis. The \u0394Eint value of 7 is stronger than NBS\u00b7\u00b7\u00b7NPy for N\u2013Br covalent bond and Br\u00b7\u00b7\u00b7N non-covalent bond are significantly different, indicating the absence of a shared-shell character are slightly larger than in 7 . This agrees with the well-known fact that the fused six-membered ring in NBP removes electron density from the five-membered ring, consequently making the bromine more electrophilic. The \u0394Eint values of ancillary interactions, for example, N\u00b7\u00b7\u00b7\u03c0(ring-centroid) and C\u2013H\u00b7\u00b7\u00b7N hydrogen bond contacts in 7, are estimated to understand their interaction strengths relative to Br\u00b7\u00b7\u00b7N motifs and C\u2013H\u00b7\u00b7\u00b7N contacts are weaker than XBs . The presence of the BCPs and bond paths of their connecting atoms are other evidences for N\u00b7\u00b7\u00b7\u03c0 and C\u2013H\u00b7\u00b7\u00b7N contacts. This suggests that weak and moderately strong HBs, that originate from donor-acceptor components' electron-rich and deficient sites, are inevitable and may contribute to the XB stabilization energy.In contrast to the same \u03c3-hole strengths of NBS and NBP, the Br\u00b7\u00b7\u00b7N halogen bonds interaction strengths of tifs see . The \u0394Ei11 and [HMTA]\u00b7[NIS]4 are larger than corresponding bromine structures, and twice the energy of C\u2013I\u00b7\u00b7\u00b7N contacts in [HMTA]\u00b7[Iodopentafluorobenzene]2 (see int and the \u03c1(r) values at the BCP of 11 are somewhat larger than [HMTA]\u00b7[NIS]4, which agrees with the additivity analysis. Complex 12 involving NISac has the largest interaction energy of all the (imide)I\u00b7\u00b7\u00b7NHMTA halogen bonds owing to larger iodine \u03c3-hole strength in NISac . The \u03c1(r) values at the BCPs of I\u00b7\u00b7\u00b7N halogen bond, similar to N\u2013I covalent indicating a degree of covalency with shared-shell character, is remarkable. This agrees with the N\u2013I and I\u00b7\u00b7\u00b7N bonds covalent character discussion in [DMAP]\u00b7[NISac] and are comparable to HB energies observed in 7. The AIM analysis reveals that C\u2013H\u00b7\u00b7\u00b7O=S and C\u2013H\u00b7\u00b7\u00b7\u03c0 interactions may contribute to the formation of I\u00b7\u00b7\u00b7NHMTA interactions.The I\u00b7\u00b7\u00b7N interaction energies of ne]2 see . The \u0394EiN-iodosaccharin even approaches the reported 3-center-4-electron halogen bonds of [(HMTA)N\u2013I\u2013N(HMTA)]+ [2.288(14) and 2.299(15) \u00c5]. The scope of halogen-bonded organic frameworks (XBOFs), previously accessed by (imide)I\u00b7\u00b7\u00b7N using a 1:4 [HMTA]:[NIS] building block, is expanded to (imide)Br\u00b7\u00b7\u00b7N halogen-bonded 1:3 [HMTA]:[NBS] and 1:4 [HMTA]:[NBS] structures. Different from (imide)I\u00b7\u00b7\u00b7N XBOFs, channel shape adaptability is achieved through HMTA tridentate and tetradentate coordination modes. DFT based MEPs provided us with important experimental insights into the nature of donor-donor and donor-acceptor interactions. Donors, such as NBS/NIS possessing \u03c3-hole and C\u2013H acidic proton values, have high probabilities to form XBOFs via (imide) X\u00b7\u00b7\u00b7N halogen bonds and orthogonal C\u2013H\u00b7\u00b7\u00b7O=C hydrogen bonds. The lack of acidic sp3 C\u2013H protons, like in NBP, encourage \u03c0-\u03c0 and other hydrogen bond interactions obstructing the formation of the desired 1:4 ratio [HMTA]:[NBP] and eventually affect XBOFs' self-assembly processes. In terms of DFT interactions energies, the (imide)N\u2013X\u00b7\u00b7\u00b7N(HMTA) halogen bonds varying from \u221211.2 to \u221212.5 kcal/mol for X = Br, and \u22128.4 to \u221229.0 kcal/mol for X = I, are stronger than corresponding (imide)N\u2013X\u00b7\u00b7\u00b7N(pyridines) halogen bonds. A comprehensive solution NMR study on [HMTA]\u00b7[N-haloimide]n complexes, optimization of crystallization conditions to synthesize XBOFs using other HMTA-imide combinations, and post-synthetic solvent exchange process of (imide)N\u2013Br\u00b7\u00b7\u00b7N XBOFs are currently under investigation in our laboratory.In summary, we investigated Y\u2013X\u00b7\u00b7\u00b7N halogen bonds in 13 X-ray crystal structures obtained from HMTA and N-haloimides. Two complexes of HMTA with iodoperfluorobenzene and 1,4-diiodotetrafluorobenzene consisting of C\u2013I\u00b7\u00b7\u00b7N halogen bonds were also prepared, and their solid-state structures were studied for comparison purposes. The Y\u2013X\u00b7\u00b7\u00b7N distances depend on the nature of Y-atom and the donor scaffold. The Br/Cl\u2013Br\u00b7\u00b7\u00b7N [2.088(3)\u22122.167(3) \u00c5] are shorter than (imide)N\u2013Br\u00b7\u00b7\u00b7N [2.371(8)\u22122.426(4) \u00c5] halogen bonds. In contrast, the I/Cl\u2013I\u00b7\u00b7\u00b7N [2.328(3)\u22122.486(5) \u00c5] tend to be longer when compared to (imide)N\u2013I\u00b7\u00b7\u00b7N [2.29(3) and 2.502(10) \u00c5] halogen bonds. The shortest I\u00b7\u00b7\u00b7N [2.29(3) \u00c5] distance between HMTA and The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/12, for X-ray crystallography, done in JYU, were prepared by GA, 12 was prepared by LG. Both JR and GP were external thesis supervisors for GA and LG, respectively. AF and AB were responsible for computational studies. KR was responsible for proofreading the final manuscript version. All authors have read and agreed to the published version of the manuscript.RP was responsible for supervision, methodology development, manuscript preparation, and SCXRD analysis. All halogen bond complexes, except The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The mol\u00adecular structure is stabilized by an O\u2014H\u22efN hydrogen bond, forming an S(6) ring motif.The title compound, C 22H18N2O2, is a Schiff base that exists in the phenol\u2013imine tautomeric form and adopts an E configuration with respect to the C=N bond. The mol\u00adecular structure is stabilized by an O\u2014H\u22efN hydrogen bond, forming an S(6) ring motif. In the crystal, pairs of C\u2014H\u22efO hydrogen bonds link the mol\u00adecules to form inversion dimers. Weak \u03c0\u2013\u03c0 stacking inter\u00adactions along the a-axis direction provide additional stabilization of the crystal structure. The mol\u00adecule is non-planar, the aromatic ring of the benzaldehyde residue being nearly perpendicular to the phenyl and 4-methyl\u00adphenol rings with dihedral angles of 88.78\u2005(13) and 82.26\u2005(14)\u00b0, respectively. A mol\u00adecular docking study between the title mol\u00adecule and the COVID-19 main protease (PDB ID: 6LU7) was performed, showing that it is a potential agent because of its affinity and ability to adhere to the active sites of the protein.The title compound, C We report here the synthesis, crystal and mol\u00adecular structure of the title compound. We have also performed a mol\u00adecular docking study to determine possible inter\u00admolecular inter\u00adactions between the COVID-19 main protease (PDB ID: 6LU7) and the title compound.Schiff bases have wide applications inter\u00adests as corrosion inhibitors ring motif, which stabilizes the mol\u00adecular structure. The di\u00adbenzyl\u00adidene hydrazine unit is approximately planar with the dihedral angle formed by the two terminal phenyl rings of 7.62\u2005(15)\u00b0. On the other hand, the mol\u00adecule is non-planar, because the C1\u2013C6 ring is nearly perpendicular to the C9\u2013C14 and C16\u2013C21 rings with dihedral angles of 88.78\u2005(13) and 82.26\u2005(14)\u00b0, respectively. The C17\u2014O2, C15\u2014N2 and C15\u2014C16 bond lengths in the mol\u00adecule are 1.359\u2005(5), 1.287\u2005(5), and 1.452\u2005(5)\u2005\u00c5, respectively. These results suggest single-bond character for C17\u2014O2 and C15\u2014C16 and double-bond character for the C15\u2014N2 bond as expected for a phenol\u2013imine structure ring motif.The asymmetric unit of the title structure contains one mol\u00adecule Fig.\u00a01, which cCg2\u22efCg3 = 3.909\u2005(2)\u2005\u00c5; Cg2 and Cg3 are the centroids of the C9\u2013C14 and C16\u2013C21 rings, respectively] that stabilize the crystal structure, forming a three-dimensional network.In the crystal, mol\u00adecules are linked by pairs of C3\u2014H3\u22efO2 hydrogen bonds, forming inversion dimers with an f Table\u00a01. There aet al., 2016et al., 2011sp2\u2014Csp2 single bond, is slightly longer than observed for the title compound [1.472\u2005(5)\u2005\u00c5]. This bond length is shorter than in NOTZIH hydrazineyl\u00adid\u00adene}-1,2-di\u00adphenyl\u00adethan-1-one was prepared by refluxing a mixture of a solution containing 2-hy\u00addroxy-5-methyl\u00adbenzaldehyde (0.02\u2005mmol) in ethanol (20\u2005mL) and a solution containing (E)-2-hydrazineyl\u00adidene-1,2-di\u00adphenyl\u00adethan-1-one (0.02\u2005mmol) in ethanol (20\u2005mL). The reaction mixture was stirred for 5\u2005h under reflux. The obtained crystalline material was washed with ethanol and dried at room temperature. Single crystals of the title compound for X-ray analysis were obtained by slow evaporation of an ethanol solution.(Uiso(H) = 1.5Ueq(O). The C-bound H atoms were positioned geometrically and refined using a riding model with C\u2014H = 0.93 and Uiso(H) = 1.2Ueq(C) for aromatic H atoms, and with C\u2014H = 0.96\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021005442/yk2152sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021005442/yk2152Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021005442/yk2152Isup3.cmlSupporting information file. DOI: 2085577CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adethanedi\u00adamine, C6H16N2, crystallizes in the monoclinic crystal system in the space group P21/c. For the investigation of the conformation, quantum chemical methods were used and for inter\u00admolecular inter\u00adactions, a Hirshfeld surface analysis was performed. N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adethanedi\u00adamine, C6H16N2, is a bidentate amine ligand commonly used in organolithium chemistry for deaggregation. Crystals were grown at 243\u2005K from n-pentane solution. The complete mol\u00adecule is generated by a crystallographic center of symmetry and the conformation of the di\u00adamine is anti\u00adperiplanar. To investigate the inter\u00admolecular inter\u00adactions, a Hirshfeld surface analysis was performed. It showed that H\u22efH inter\u00adactions dominate with a contact percentage of 92.3%.The title compound N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adethanedi\u00adamine consists of two tertiary amine groups linked by an ethyl\u00adene bridge. It can be used in cross-coupling or in olefin polymerization reactions where, e.g., a complex between di\u00admethyl\u00adnickel and TMEDA is used as a catalyst \u2212x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z] = 180.0\u00b0 by symmetry. The conformations of the C1\u2014N1\u2014C3\u2014C3i and C2\u2014N1\u2014C3\u2014C3i groupings are anti [torsion angle = 167.33\u2005(6)\u00b0] and gauche [\u201371.17\u2005(8)\u00b0], respectively.Compound et al., 2013Gaussian 16 -tris\u00ad(tetra\u00admethyl\u00adethylenedi\u00adamine\u00adlithium)methyl\u00adthorium(IV) tetra\u00admethyl\u00adethylenedi\u00adamine -({[(S)-2-(meth\u00adoxy\u00admeth\u00adyl)pyrrolidin-1-yl]meth\u00adyl}di\u00admeth\u00adyl\u00adsil\u00adyl)(phen\u00adyl)meth\u00adyl]lithium N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adethane\u00addi\u00adamine was purchased by Sigma-Aldrich and was used without further purification. A solution of TMEDA (0.5\u2005mmol) in n-pentane (1\u2005ml) was prepared at 243\u2005K and 1 crystallized in the form of colorless blocks.Uiso(H) = 1.2Ueq(C) for CH2 and CH hydrogen atoms and Uiso(H) = 1.5Ueq(C) for CH3 hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021012457/hb8002sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021012457/hb8002Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021012457/hb8002Isup3.cmlSupporting information file. DOI: 2123810CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III complex with a multidentate NSO-containing mixed-ligand \u2212 2-hy\u00addroxy-3-meth\u00adoxy\u00adbenzaldehyde thio\u00adsemicarbazone \u2013 is reported.The synthesis, crystal structure and spectroscopic characterization of the novel and, according to our knowledge the first to be obtained in crystalline form, Co III complex, bis\u00ad[bis\u00adcobalt(III)] di\u00adthio\u00adnate\u2013dimethylformamide\u2013methanol (1/4/3), [Co(C9H10N3O2S)2]2(S2O6)\u00b74C3H7NO\u00b73CH3OH, with monodeprotonated 2-hy\u00addroxy-3-meth\u00adoxy\u00adbenzaldehyde thio\u00adsemicarbazone as ligands crystallizes in the space group PL2]+ cations, one di\u00adthio\u00adnate anion (S2O6)2\u2212 as counter-anion and seven solvate mol\u00adecules (four di\u00admethyl\u00admethanamide and three methanol). Each CoIII ion has a moderately distorted octa\u00adhedral S2N2O2 geometry. In the crystal, the components are linked by numerous N\u2014H\u22efO and O\u2014H\u22efO contacts.The title Co Thus, the coordination geometry around each CoIII ion can be described as moderately distorted octa\u00adhedral with an S2N2O2 coordination sphere with N,O,N and S atoms in the equatorial plane and O and S atoms in the apical positions.The title complex crystallizes in the triclinic space group it Fig.\u00a01 consistset al., 2013II complexes with related semicarbazone ligands to those in an analogous chromium complex with a similar ligand approaches the standard C=S double-bond value and differs only slightly from the distance observed in the corresponding neutral ligand [1.688\u2005\u00c5 in BIZYAL and twisted, as defined by the dihedral angles of 83.42\u2005(7)\u00b0 between the mean planes of atoms O1/C1/C6/C8/N1/N2/C9/S1 and O3/C10/C15/C17/N4/N5/C18/S2 around Co1, and 86.3\u2005(1)\u00b0 between the mean planes of atoms O7/C28/C33/C35/N10/N11/C36/S4 and O5/C19/C24/C26/N10/N8/C27/S3 around Co2.The ligands coordinated to the CoThe solid-state organization of the complex can be described as an insertion of the anions and solvent mol\u00adecules within the crystallographically independent complexes Fig.\u00a02. In the A\u22efO8, N2\u2014H2\u22efO3 and N2\u2014H2\u22efO4 are contacts between ligands through the nitro\u00adgen of the secondary amino group and meth\u00adoxy group oxygen;N8\u2014H8A\u22efO11 and N12\u2014H12B\u22efO9 are contacts between the nitro\u00adgen of the secondary and primary amino groups of the ligands and oxygen atoms of the S2O6 anions .N3\u2014H32O6)2\u2212 anions act as a multiple-acceptor species for N,O donor atoms of neighboring complexes (by N\u2014H\u22efO inter\u00adactions) and methanol solvent mol\u00adecules (by O\u2014H\u22efO contacts). The oxygen atoms (O16) of the di\u00admethyl\u00admethanamide mol\u00adecules bridge adjacent cationic complexes of the ligand in 1ml of di\u00admethyl\u00adformamide and 1ml of chloro\u00adform. Dark-brown crystals of the title compound, suitable for X-ray analysis, were formed within a few days (yield: 60%).The title compound was prepared according to a previously published procedure due to aromatic =C\u2014H stretching at 3000\u20133100\u2005cm\u22121, the aromatic ring vibrations in the 1600\u20131400\u2005cm\u22121 region, weak absorption band at 738\u2005cm\u22121 due to \u03c5(C\u2014S) vibrations and the characteristic peak at 1608\u2005cm\u22121 assigned to azomethine \u03c5(C=N) group. The weak band at 3308\u2005cm\u22121 can be assigned to the N\u2014H group vibrations. All these data are in good agreement with literature data : C 38.19; N 14.33; H 5.16%. Found: C 38.21; N 14.40; H 5.21%.The IR spectrum of the title compound (as KBr pellets) is consistent with the above structural data. In the range 4000\u2013400\u2005cmII di\u00adthio\u00adnate used in this work was prepared by mixing aqueous solutions containing stoichiometric amounts of cobalt sulfate and BaS2O6\u00b72H2O. The white precipitate of BaSO4 was removed by filtration and the solution containing the metal di\u00adthio\u00adnate was evaporated to a small volume on a rotary evaporator and then cooled for crystallization. BaS2O6\u00b72H2O was prepared using the method described by Pfanstiel (1946The Cotiel 1946.Uiso(H) = 1.2Ueq(C) for aromatic CH and Uiso(H) = 1.5Ueq(C) for methyl groups. The H atoms of the NH and OH groups were also placed at calculated position using the corresponding AFIX instruction with Uiso(H) = 1.2Ueq(N) for NH/NH2 and Uiso(H) = 1.5Ueq(O) for OH hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021010616/tx2043sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021010616/tx2043Isup2.hklStructure factors: contains datablock(s) I. DOI: 2115486CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title thio\u00adurea derivative adopts a U-shaped conformation, which incorporates an intra\u00admolecular amine-N\u2014H\u22efN(imine) hydrogen bond. In the mol\u00adecular packing, supra\u00admolecular chains are formed through hydroxyl-O\u2014H\u22efS(thione) and amine-N\u2014H\u22efO hydrogen bonding. 17H19N3OS, adopts a U-shaped conformation with the dihedral angle between the terminal aromatic rings being 73.64\u2005(5)\u00b0. The major twist in the mol\u00adecule occurs about the ethane bond with the Ci\u2014Ce\u2014Ce\u2014Cb torsion angle being \u221278.12\u2005(18)\u00b0; i = imine, e = ethane and b = benzene. The configuration about the imine bond is E, the N-bound H atoms lie on opposite sides of the mol\u00adecule and an intra\u00admolecular amine-N\u2014H\u22efN(imine) hydrogen bond is evident. In the mol\u00adecular packing, hydroxyl-O\u2014H\u22efS(thione) and amine-N\u2014H\u22efO hydrogen bonding feature within a linear, supra\u00admolecular chain. The chains are connected into a layer in the ab plane by a combination of methyl\u00adene-C\u2014H\u22efS(thione), methyl\u00adene-C\u2014H\u22efO(hydrox\u00adyl), methyl-C\u2014H\u22ef\u03c0(phen\u00adyl) and phenyl-C\u2014H\u22ef\u03c0(hy\u00addroxy\u00adbenzene) inter\u00adactions. The layers stack without directional inter\u00adactions between them. The analysis of the calculated Hirshfeld surface highlights the presence of weak methyl-C\u2014H\u22efO(hydrox\u00adyl) and H\u22efH inter\u00adactions in the inter-layer region. Computational chemistry indicates that dispersion energy is the major contributor to the overall stabilization of the mol\u00adecular packing.The title thio\u00adurea derivative, C Globally, both aromatic residues lie to the same side of the mol\u00adecule so that it has a U-shaped conformation.The mol\u00adecular structure of (I)E. The N-bound H atoms lie to opposite sides of the mol\u00adecule, a conformation that allows for the formation of an intra\u00admolecular amine-N\u2014H\u22efN(imine) hydrogen bond, Table\u00a01The C1\u2014S1 bond length is 1.6910\u2005(15)\u2005\u00c5, the C1\u2014N1 bond [1.340\u2005(2)\u2005\u00c5] is marginally shorter than the C1\u2014N2 [1.356\u2005(2)\u2005\u00c5] bond, the formally C8\u2014N3 double bond is 1.284\u2005(2)\u2005\u00c5 and N2\u2014N3 is 1.3857\u2005(18)\u2005\u00c5. These values, coupled with the observed planarity in this region of the mol\u00adecule, is suggestive of some delocalization of \u03c0-electron density over this residue. The configuration about the C8=N3 imine bond is a). They are connected into a supra\u00admolecular layer parallel to the c axis via methyl\u00adene-C\u2014H\u22efS(thione) and methyl\u00adene-C\u2014H\u22efO(hydrox\u00adyl) inter\u00adactions as well as methyl-C\u2014H\u22ef\u03c0(phen\u00adyl) and phenyl-C\u2014H\u22ef\u03c0(hy\u00addroxy\u00adbenzene) contacts, Table\u00a01b). The layers thus formed are two mol\u00adecules thick and stack along the c-axis direction without directional inter\u00adactions between them, Fig.\u00a02c). Finally, as indicated in Fig.\u00a02b) and (c), the supra\u00admolecular connectivity brings two sulfur atoms into close proximity, with an S1\u22efS1i separation of 3.3534\u2005(6)\u2005\u00c5, cf. the sum of the van der Waals radii of 3.60\u2005\u00c5 , i.e. near the amine-H2N and thione-S1 atoms, correspond to the amine-N2\u2014H2N\u22efO1(hydrox\u00adyl), hydroxyl-O1\u2014H1O\u22efS1(thione) hydrogen bonds and the thione-S1\u22efS1(thione) short contact; these and other short contacts calculated using Crystal Explorer 17 are collated in Table\u00a02b), where the positive electrostatic potential (blue) and negative electrostatic potential (red) regions are observed around the amine-H2N and thione-S1 atoms, respectively. The faint red spots appearing near the thione-S1, hydroxyl-O1 and methyl\u00adene-H11A and H11B atoms and phenyl-C6\u2014H6\u22ef\u03c0 inter\u00adactions are shown as faint red spots on the dnorm surface in Fig.\u00a05a) and as two distinctive orange \u2018potholes\u2019 on the shape-index-mapped over Hirshfeld surface in Fig.\u00a05b). It is noted that the phenyl-C4\u2014H4\u22ef\u03c0 inter\u00adaction, Table\u00a01dnorm-mapped Hirshfeld surface. However, this inter\u00adaction clearly shows up as an orange \u2018pothole\u2019 on the shape-index-mapped Hirshfeld surface in Fig.\u00a06The bright-red spots on the Hirshfeld surface mapped over ms Fig.\u00a04 correspoa) and those delineated into H\u22efH, H\u22efC/C\u22efH, H\u22efS/S\u22efH and H\u22efO/O\u22efH contacts are illustrated in Fig.\u00a07b)\u2013(e), respectively. The percentage contributions to the Hirshfeld surface of (I)de = di \u223c2.1\u2005\u00c5, Fig.\u00a07b), corresponds to the intra-layer H1O\u22efH2N contact listed in Table\u00a02c), reflecting the significant C\u2014H\u22ef\u03c0 inter\u00adactions evinced in the packing analysis, Table\u00a01de = di \u2243 2.2\u2005\u00c5 in Fig.\u00a07d) and (e), respectively. The contributions from the other six inter\u00adatomic contacts summarized in Table\u00a03The overall two-dimensional fingerprint plot computed for (I)Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energy components and Erep (12.2\u2005kJ\u2005mol\u22121) terms and having a total energy of 11.7\u2005kJ\u2005mol\u22121 is non-attractive.The energy frameworks were calculated for (I)Edis component. The greatest stabilization energy in the inter-layer region arises from methyl-C9\u2014H9B\u22efO1(hydrox\u00adyl) and methyl\u00adene-H10\u22efH16(hy\u00addroxy\u00adbenzene) inter\u00adactions , which sum to \u221230.7\u2005kJ\u2005mol\u22121. Generally, the long-range H\u22efH contacts are the major inter\u00adactions stabilizing the mol\u00adecules within the inter-layer region.The stabilization energies in the inter-layer region are dominated by the a axis direction are shown in Fig.\u00a08Eele of all pairwise inter\u00adactions sum to \u2212145.4\u2005kJ\u2005mol\u22121, while the Edis totals \u2212342.1\u2005kJ\u2005mol\u22121.Views of the energy framework diagrams down the via an ethane link. Each of these is a N-methyl species, i.e. MeN(H)C(=S)N(H)N=C(Me)CH2CH2Ar, one with Ar = phenyl torsion angle = \u221262.76\u2005(16)\u00b0], a conformation stabilized, at least in part, by an intra\u00admolecular amine-N\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adaction. By contrast, in the species with Ar = 4-meth\u00adoxy\u00adbenzene, the mol\u00adecule is close to planar as indicated by the Ci\u2014Cm\u2014Cm\u2014Cp torsion angles of 177.51\u2005(12) and \u2212175.80\u2005(12)\u00b0, respectively, for the two independent molecules comprising the asymmetric unit. Thus, to a first approximation, the conformation observed in (I)In the crystallographic literature, there are two precedents for mol\u00adecules related to (I)in vacuo. Single crystals were grown at room temperature in mixed solvents of dimethyformamide and aceto\u00adnitrile (1:2 v/v) by slow evaporation. 1H NMR : \u03b4 9.14 , 8.59 , 7.59 , 7.37 , 7.22 , 7.03 , 6.76 , 5.46 , 2.83 , 2.61 , 1.90 . 13C NMR : \u03b4 176.22, 154.16, 152.02, 137.93, 132.72, 129.39, 128.84, 126.16, 124.39, 115.57, 40.38, 31.58, 16.19.4-Phenyl-3-thio\u00adsemicarbazide (10\u2005mmol) dissolved in hot absolute ethanol (50\u2005ml) was combined with 4-(4-hy\u00addroxy\u00adphen\u00adyl)-2-butanone (10\u2005mmol), dissolved in hot absolute ethanol (50\u2005ml) with a few drops of concentrated hydro\u00adchloric acid added as catalyst. The mixture was heated (348\u2005K) and stirred for about 30\u2005min. The mixture was allowed to cool to room temperature while stirring. The white precipitate was filtered, washed with cold ethanol and dried Uiso(H) set to 1.2\u20131.5Ueq(C). The O-bound and N-bound H atoms were located in a difference-Fourier map but were refined with O\u2014H = 0.84\u00b10.01 and N\u2014H = 0.88\u00b10.01\u2005\u00c5 distance restraints, respectively, and with Uiso(H) set to 1.5Ueq(O) and 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989021006666/hb7978sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989021006666/hb7978Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021006666/hb7978Isup3.cmlSupporting information file. DOI: 2092413CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Further, cleavage of the [Fe\u2010S\u2010Fe] unit by CS2 is presented.A [Fe\u2010S\u2010Fe] subunit with a single sulfide bridging two low\u2010coordinate iron ions is the supposed active site of the iron\u2010molybdenum co\u2010factor (FeMoco) of nitrogenase. Here we report a dinuclear monosulfido bridged diiron(II) complex with a similar complex geometry that can be oxidized stepwise to diiron(II/III) and diiron(III/III) complexes while retaining the [Fe\u2010S\u2010Fe] core. The series of complexes has been characterized crystallographically, and electronic structures have been studied using, inter alia, A dinuclear monosulfido bridged diiron(II) complex is reported that can be oxidized stepwise to diiron(II/III) and diiron(III/III) complexes while retaining the [Fe\u2010S\u2010Fe] core. This unprecedented series of low\u2010coordinate complexes has been thoroughly characterized using inter alia, 57Fe M\u00f6ssbauer spectroscopy and SQUID magnetometry. The reaction takes place at the iron\u2010molybdenum co\u2010factor (FeMoco), which constitutes a ligated [MoFe7S9C] unit.Nitrogenase (N2+ complex with an unsupported monosulfide bridge via the reaction of a two\u2010coordinate iron(I) silylamide with elemental sulfur. Subsequent oxidation leads to the first example of a mixed valent [2Fe\u20101S]3+ and an \u201call ferric\u201d [2Fe\u20101S]4+ form. The series of complexes was examined with respect to their spectroscopic and physical properties. The initial [2Fe\u20101S]2+ complex was further subjected to a variety of small molecule substrates that are transformed by the N2\u2010ase, however showing only a limited reactivity or stability. Most notably, its reaction with CS2 led to rupture of the Fe\u2010S\u2010Fe motif and formation of a mononuclear iron(II) thiocarbonate complex, revealing the structural lability of the [Fe\u2010S\u2010Fe] unit.This unit is assumed to open up during substrate turnover, as experimentally shown by displacement of the sulfide unit by CO or selenide.2, 6 A2] (L=\u2010N(Dipp)SiMe3, Dipp=2,6\u2010diisopropylphenyl), 1,2O with 1/16 S8 for 16\u2005h led to the formation of a colorless solid. X\u2010Ray diffraction analysis of suitable single crystals showed the formation of the dinuclear complex (K{18c6})2[(FeL2)2(\u03bc\u2010S)], 2 (Scheme\u2005The reaction of a suspension of K{18c6}FeL (L=\u2010N(Di2 features two three\u2010coordinate iron(II) ions bridged by a single sulfide in a nearly linear fashion (Fe\u2010S\u2010Fe 167.78(2)\u00b0) with slightly different Fe\u2212S distances of 2.2400(5)\u2005\u00c5 and 2.337(5)\u2005\u00c5 complex bearing an unsupported sulfide bridge.2 can be adjusted accordingly. The cyclic voltammogram of 2 in THF showed two quasi\u2010reversible one\u2010electron oxidation processes at E1/2=\u22121.55\u2005V and \u22120.55\u2005V 2] (monoanionic ligand L\u2212=nacnac) reported by Driess and co\u2010workers features redox events in THF at E1/2=\u22121.45\u2005V for the [2Fe\u20102S]2+/1+ and E1/2=\u22122.55\u2005V (\u0394E1/2=1.10\u2005V) for the [2Fe\u20102S]1+/0 redox couple.2(\u03bc\u2010S)2]2\u2212 2+ core in MeCN occurred at E1/2=\u22121.14\u2005V ([2Fe\u20102S]2+/1+) and E1/2=\u22122.10\u2005V .2 (\u0394E1/2=0.98\u2005V). In contrast, the positions of the redox events of these bis(\u03bc\u2010sulfido) complexes are shifted by around 0.9\u2005V or 0.6\u2005V to lower potentials, respectively, reflecting the additional sulfide ligation and higher coordination number of the iron ions in the two previously reported [2Fe\u20102S] systems.[The related bis(\u03bc\u2010sulfido) diferric complex [(LFe)2. Treatment of 2 with one equivalent of AgOTf in Et2O yielded reddish K{18c6}[(L2Fe)2(\u03bc\u2010S)], 3 led to a colour change to dark green, and the neutral complex [(L2Fe)2(\u03bc\u2010S)], 4, was obtained from a saturated pentane solution.Given the electrochemical data we attempted the chemical oxidation of 3 Scheme\u2005. Treatme3 and 4 are shown in Table\u20052, complexes\u20053 and 4 retain both a more or less linear Fe\u2010S\u2010Fe core. A gradual decrease of the Fe\u2212S and Fe\u2212N bond lengths was observed upon oxidation due to the contraction of the ionic radii of the metal ions. The structural features found for both iron ions are largely identical in 3 and 4, which prohibited an assignment of localized oxidation states for mixed\u2010valent 3 on a structural level.Important structural parameters of 2 showed no absorption beyond 400\u2005nm , 470\u2005nm (\u03f5=9540\u2005L\u2009mol\u22121\u2009cm) and 700\u2005nm (\u03f5=2270\u2005L\u2009mol\u22121\u2009cm), and it is tempting to assign the low\u2010energy absorption to an intervalence charge transfer (IVCT) transition. For 4 just one pronounced band at 430\u2005nm (\u03f5=5710\u2005L\u2009mol\u22121\u2009cm) was observed. Zero field 57Fe M\u00f6ssbauer spectroscopy complex.\u03b4=0.86\u2005mm\u2009s\u22121 and |\u0394EQ|=0.58\u2005mm\u2009s\u22121), which can be explained by the weaker donor strength of the silylamide ligands as well as a less distorted trigonal planar ligand arrangement.4 is represented by a doublet with \u03b4=0.29\u2005mm\u2009s\u22121 and |\u0394EQ|=3.70\u2005mm\u2009s\u22121 indicating the presence of high\u2010spin iron(III) ions. The spectrum of the mixed valent complex\u20053, recorded at 7\u2005K, showed two doublets with \u03b4=0.36\u2005mm\u2009s\u22121 (|\u0394EQ|=3.70\u2005mm\u2009s\u22121) and \u03b4=0.57\u2005mm\u2009s\u22121 (|\u0394EQ|=0.71\u2005mm\u2009s\u22121). This evidences distinguishable iron(II/III) positions in solid 3 on the 57Fe M\u00f6ssbauer timescale at 7\u2005K, whereas the smaller separation in the isomer shifts (\u0394\u03b4(3)=0.21\u2005mm\u2009s\u22121 vs. \u0394\u03b4(2/4)=0.30\u2005mm\u2009s\u22121) is indicative of some degree of valence delocalisation. Given the lack of literature precedence of the three\u2010coordinate \u03bc\u2010sulfido complexes\u20053 and 4 their M\u00f6ssbauer spectroscopic features are compared best to low coordinate iron complexes bearing a [2Fe\u20102S] motif in the same oxidation states.[With this unprecedented series of low\u2010coordinate 2Fe\u20101S] complexes in three different oxidation states in hand, their spectroscopic and electronic features were studied in detail. UV/Vis spectroscopic examination of m Figure\u2005, which iy Figure\u2005 revealedS complex2\u20134 in solid state were obtained by SQUID measurements (S=2) ions with a S=0 ground state. The coupling constant was determined to be J=\u221253\u2005cm\u22121 using \u0124=\u22122JSA\u22c5SB with g1=g2=2.01.Additional insights into the electronic situation of s Figure\u2005\u2013F. 2 exh3 showed at 295\u2005K a significantly lower \u03c7T value of 0.96\u2005cm3\u2009mol\u22121\u2009K which decreased to 0.44\u2005cm3\u2009mol\u22121\u2009K at 80\u2005K with a further drop to 0.38\u2005cm3\u2009mol\u22121\u2009K below 20\u2005K, which implies a ground state of S=1/2. The antiferromagnetic coupling is stronger with J=\u2212115\u2005cm\u22121 . For the all\u2010ferric compound\u20054 a similar value J=\u2212104\u2005cm\u22121 was observed with \u03c7T=1.3\u2005cm3\u22c5mol\u22121\u22c5K at 300\u2005K that decreased linearly to ca. 0.2\u2005cm3\u2009mol\u22121\u2009K below 50\u2005K due to a S=0 ground state. The differences in exchange coupling can be explained using a simplified orbital scheme under assumption of an idealized C2V symmetric ligand environment for each iron atom . Upon oxidation, electrons are removed from the lowest\u2010lying, co\u2010parallel dxy/dx2\u2010y2 orbitals, which have no impact onto the exchange mechanism. As such the variation in J values for 2\u20134 can be mainly attributed to differences in Fe\u22c5\u22c5\u22c5Fe distances, with different superexchange contributions due to changes in Fe\u2010(\u03bc\u2010S) covalency likely playing a further role. A significant stabilization of the antiferromagnetically coupled ground state upon oxidation from the diiron(II) to the mixed\u2010valent iron(II)/iron(III) and diiron(III) states was observed for the series of complexes [(LFe)2(\u03bc\u2010S)2]4\u2212/3\u2212/2\u2212 (L2\u2212=bis(benzimidazolato))., 5 thiocarbonate complex\u20057, which could alternatively be obtained via the oxidation of 6 by silver triflate. For the neutral complex\u20054 the reaction with CS2 remained inconclusive. The observation of facile Fe\u2013S bond cleavages suggests a rather weak FeII\u2013S interaction. The displacement of an iron(II) ion by other Lewis acids has possible implications for the situation found in the FeMo cofactor where cleavage of the belt Fe\u2010S\u2010Fe unit is discussed during substrate turnover using the local Lewis acid/base properties of the surroundings of the enzyme pocket.2 into a [Fe\u2010S\u2010Fe] function also reveals how CS2 might act as an inhibitor of nitrogenase FeMoco (and other iron\u2010sulfur clusters) which is thought to proceed by blocking of coordination sites3+ complex localized valence states in solid state at low temperatures. Magnetic measurements revealed for the diferrous [2Fe\u20101S]2+ a moderate antiferromagnetic coupling which becomes significantly enhanced for the [2Fe\u20101S]3+ and [2Fe\u20101S]4+ compounds. Reactivity studies on these complexes towards different nitrogenase relevant substrates revealed for CS2 the facile cleavage of the Fe\u2010S\u2010Fe unit. This led to the formation of an iron thiocarbonate which may suggest a possible inhibitory mechanism of CS2 with respect to the reactivity of FeMoco and related Fe/S clusters.We have synthesized a unique series of low\u2010coordinate [Fe\u2010S\u2010Fe] complexes in three oxidation states which resembles a Fe\u2010S\u2010Fe belt unit in the iron/sulfur/molybdenum co\u2010factor of the nitrogenase enzyme. These complexes were characterized for their magnetic and spectroscopic properties. 2] was synthesized according to the literature procedure. For details concernining data acquisition of solution and solid\u2010state analyses , see the Supporting Information.General considerations: All manipulations were carried out in a glovebox, or using Schlenk\u2010type techniques under a dry argon atmosphere. Used solvents were dried by continuous distillation over sodium metal for several days, degassed via three freeze\u2010pump cycles and stored over molecular sieves 4\u2005\u00c5. K{18c6}[FeL2[(FeL2)2(\u03bc\u2010S)] (2)[K{18\u2009c6}]: [K{18c6}][FeL2], 1, was suspended in 5\u2005mL of Et2O. The slow addition of elemental sulfur led to an immediate colour change of the solution from red to brown and the precipitation of a pale yellow solid. Decanting off the supernatant, washing the residue with 2\u00d75\u2005mL of pentane and drying under reduced pressure afforded the crude product as a pale yellow crystalline solid. Recrystallization in THF/ pentane at \u221235\u2009\u00b0C led to colourless crystals of 2 , suitable for X\u2010ray diffraction. 1H\u2010NMR : \u03b4=14.23, 9.30, 3.50, 1.96, \u22120.94, \u22121.70\u2005ppm. Evans: : eff\u03bc=3.98\u2005\u03bcB.FT\u2010IR (ATR): (cm\u22121): v\u0304=2896 (w), 1418 (w), 1352 (w), 1314 (w), 1235 (m), 1192 (w), 1103 (vs.), 962 (w), 907 (s), 835 (vs.), 775 (s), 664 (w), 529 (w), 423 (m). CHNS: calc. (C84H152Fe2K2N4O12Si4S 1744.44\u2005g\u2009mol\u22121): C 57.84 H 8.78 N 3.21 S 1.84 found: C 57.93 H 8.69 N 3.58 S 1.36.2)2(\u03bc\u2010S)] (3)[K{18\u2009c6}][(FeL: [K{18c6}]2[(FeL2)2(\u03bc\u2010S)], 2, was suspended in 5\u2005mL of Et2O. Upon the addition of AgOTf the pale yellow suspension turned into a red solution with beginning precipitation of a dark solid . After stirring for 2\u2005hours, the mixture was filtered, the residue washed with 2\u00d73\u2005mL Et2O and the combined filtrates were layered with 5\u2005mL of pentane. Storing the solution at \u221235\u2009\u00b0C for several days yielded to a dark red crystalline solid, suitable for X\u2010ray diffraction analysis. Decanting off the supernatant, washing the residue with 2\u00d75\u2005mL of pentane and drying under reduced pressure afforded [K{18c6}][(FeL2)2(\u03bc\u2010S)], 3, as a dark red crystalline solid . The aforementioned procedure to synthesize 3 leads to a pure product according to elemental analysis (vide infra). To obtain a magnetically pure sample several recrystallization steps in Et2O/pentane were required, which led to a decrease of the yield to less than 10\u2009%. 1H\u2010NMR: : \u03b4=14.58, 14.10, 3.26, 3.07, \u22120.92, \u22122.01\u2005ppm. Evans: : eff\u03bc=3.70\u2005\u03bcB. FT\u2010IR (ATR): (cm\u22121): v\u0304=2954 (w), 1456 (w), 1421 (m), 1353 (w), 1309 (w), 1232 (s), 1179 (s), 1101 (vs.), 961 (m), 896 (m), 832 (vs.), 781 (vs.), 733 (s), 673 (s), 637 (w), 541 (m), 434 (s). UV/VIS (THF): \u03bb/ nm (\u03f5/ L\u2009mol\u22121\u2009cm)=380 (11\u2009800), 470 (9540), 700 (2270). CHNS: calc. (C72H128Fe2K1N4O6Si4S 1441.03\u2005g\u2009mol\u22121): C 60.01 H 8.95 N 3.89 S 2.22 found: C 59.66 H 8.64 N 4.07 S 1.51.2)2(\u03bc\u2010S)] (4)[(FeL: [K{18c6}]2[(FeL2)2(\u03bc\u2010S)], 2, was suspended in 5\u2005mL of Et2O. Upon the addition of AgOTf the pale yellow suspension turned into a dark green solution and a dark precipitate. After stirring for 2\u2005hours, the mixture was filtered, the residue washed 2 times with 3\u2005mL of Et2O and the combined filtrates were dried in vacuo. The residue was extracted with 5\u2005mL of pentane. The solution was concentrated to 0.5\u2005mL in vacuo and stored at \u221230\u2009\u00b0C for several days. This resulted in the deposition of a dark green crystalline solid, suitable for X\u2010ray diffraction. Decanting off the supernatant and drying under reduced pressure afforded [(FeL2)2(\u03bc\u2010S)], 4, as a dark green crystalline solid . 1H\u2010NMR: : \u03b4=64.37, 34.82, 23.62, \u22120.89, \u221226.88\u2005ppm. Evans: : eff\u03bc=7.26\u2005\u03bcB. FT\u2010IR (ATR): (cm\u22121): v\u0304=2958 (m), 1423 (w), 1360 (w), 1310 (w), 1242 (s), 1169 (m), 1100 (s), 1024 (s), 908 (m), 826 (vs.), 783 (vs.), 728 (vs.), 678 (s), 632 (m), 535 (s), 436 (s). UV/VIS (THF): \u03bb/ nm (\u03f5/ L\u2009mol\u22121\u2009cm)=430 (5710). CHNS: calc. (C60H104Fe2N4Si4S 1137.61\u2005g\u2009mol\u22121): C 63.35 H 9.22 N 4.94 S 2.83 found: C 63.97 H 8.75 N 5.42 S 2.65.2[L2FeII(\u03b72\u2010CS3)] (5)[K(18\u2010crown\u20106]: [K{18c6}]2[(FeL2)2(\u03bc\u2010S)], 2, was dissolved in 2\u2005mL of THF. The slow addition of CS2 led to a colour change of the solution from brown to clear orange. After stirring for 2 hours, the mixture was filtered and the filtrate layered by 20\u2005mL of pentane. Storing the solution at \u221235\u2009\u00b0C yielded to the precipitation of orange crystals, suitable for X\u2010ray diffraction. Decanting off the supernatant, washing of the residue with 2\u00d75\u2005mL of pentane and drying in vacuo afforded 5 as a dark orange crystalline solid . 1H\u2010NMR: : no identifiable signals besides for K{18c6}. FT\u2010IR (ATR): (cm\u22121): v\u0304=2954 (m), 2892 (m), 1453 (m), 1422 (m), 1350 (m), 1311 (w), 1234 (s), 1188 (m), 1104 (vs.), 1054 (m), 961 (s), 920 (s), 881 (m), 834 (vs.), 777 (s), 744 (m), 664 (m), 533 (m), 434 (m), 407 (w). CHN: calc. (C55H101Fe2K2N2O12Si2S3\u22c5THF 1268.81\u2005g\u2009mol\u22121): C 52.89 H 8.19 N 2.09 S 7.18 found: C 52.39 H 7.91 N 2.60 S 6.73.2[L2FeIII(\u03b72\u2010CS3)] (6)[K(18\u2009c6]: [K(18c6]2[L2FeII(\u03b72\u2010CS3)], 5, was dissolved in 2\u2005mL of THF. Upon the addition of AgOTf the solution turned dark red and the precipitation of a grey solid was observable. After stirring for 2\u2005hours, the mixture was filtered and the filtrate was layered with 2\u2005mL of pentane. Storing the solution at \u221235\u2009\u00b0C for several days led to the precipitation of dark red crystals, suitable for X\u2010ray diffraction. Decanting off the supernatant, washing of the residue with 2\u00d75\u2005mL of pentane and drying in vacuo afforded 6 as a dark red crystalline solid. K(18c6)OTf is the major side product of the reaction. As it has almost the same solubility as 6 in Et2O and THF, it was impossible to obtain an analytically pure sample of 6 upon recrystallization. Therefore 6 could only be characterized by X\u2010ray diffraction. It exhibits no identifiable 1H\u2005NMR spectroscopic signature.2), 2048164 (3), 2048165 (4), 2048166 (5) and 2048167 (6)Deposition Numbers 2048098 should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "Post-etch cleaning and silane application have been proven to increase bond strength, however, this step varies for each material [All-ceramic restorations currently dominate the market of indirect restorative materials due to their biocompatibility, longevity and superior aesthetics . Silica-material . The aimG1 BIS-Silane ; G2 ESPE Sil Silane Coupling Agent ; G3 Monobond Plus . Each was then divided into two subgroups, according to the surface conditioning time: T1 (1\u2009min.) or T2 (5\u2009min.). Each block was acid etched (HF 9.5% \u2212 1\u2009min), post-etching cleaned and silanized. Heat treatment was carried out at 100\u2009\u00b0C (1\u2009min.). Then a thin layer of Optibond FL (Kerr) adhesive was applied and each block was adhered to pre-heated resin at 55\u2009\u00b0C. The samples were light cured for 40\u2009s on each side (1200\u2009mW/cm2). Samples were sectioned into microspecimens (1\u2009\u00b1\u20090.2\u2009mm2) that were subjected to aging . The microspecimens were tested in tension at a crosshead speed of 0.5\u2009mm/min, until they debonded. Data analysis was carried out by a two-way ANOVA, at a significance level of 5%.Ten leucite reinforced glass ceramic blocks (IPS Empress CAD LT BL4/C 14) were divided into equal halves. Of the samples obtained, 6 were randomly divided into three groups according to the silane used: p\u2009<\u2009.001). Monobond Plus registered the lowest mean \u00b5TBS value (G3T1\u2009\u2212\u200918.5\u2009\u00b1\u20097.3\u2009MPa) and (G3T2\u2009\u2212\u200917.3\u2009\u00b1\u20095.8\u2009MPa). The type of silane coupling agent has shown to have a significant influence on the microtensile bond strength .The group featuring BIS-Silane with longer application time (G1T2) presented a mean \u00b5TBS value (32.4\u2009\u00b1\u200919.6\u2009MPa) significantly higher to all other groups (Some authors have previously suggested that silanization could benefit from longer application times, but seldom research has been found featuring this variation protocol ,4. Silan"} +{"text": "Additionally, the supra\u00admolecular inter\u00adactions were determined by Hirshfeld surface analysis to investigate the influence of these contacts on the crystal packing. 17H30NSi+\u00b7C2H5O4S\u2212, belongs to the class of a-amino\u00adsilanes and was synthesized by the alkyl\u00adation of 1-[(benzyl\u00addimethyl\u00adsil\u00adyl)meth\u00adyl]piperidine using diethyl sulfate. This achiral salt crystallizes in the chiral space group P21. One of the Si\u2014C bonds in the cation is unusually long [1.9075\u2005(12)\u2005\u00c5], which correlates with the adjacent quaternary N+ atom and was verified by quantum chemical calculations. In the crystal, the components are linked by weak C\u2014H\u22efO hydrogen bonds: a Hirshfeld surface analysis was performed to further investigate these inter\u00admolecular inter\u00adactions and their effects on the crystal packing.The title mol\u00adecular salt, C Consequently, the \u03c0-character of the Si\u2014C bond is more pronounced, leading to an elongation of the bond. Thus, a selective cleavage of the amino functionality due to the elongated Si\u2014C bond is also conceivable , the title compound, represents a compound that could lead to an extension of the aforementioned Si\u2014C bond to the nitro\u00adgen atom via the quaternary ammonium cation. Structural studies concerning this type of compound should better elucidate the reactivity as well as selectivity of Si\u2014C cleavages of the benzyl-substituted \u03b1-amino\u00adsilanes.Several derivates of these \u03b1-piperdino\u00adbenzyl\u00adsilanes have been synthesized by our research group: 1-[(benzyl\u00addimethyl\u00adsil\u00adyl)meth\u00adyl]-1-ethyl\u00adpiperidin-1-ium ethane\u00adsulfonate symmetry. The chiral space group indicates that the achiral compound in the elementary cell is packed chirally; the Flack absolute structure parameter amounts to \u22120.005\u2005(6) \u00b0 (C7\u2014Si1\u2014C10) and the largest angle of 114.32\u2005(7)\u00b0 (C8\u2014Si1\u2014C10). This geometric distortion has been observed in many complex substituted silicon compounds and depends on the substituents to 167\u2005(2)\u00b0 (C3\u2014H3\u22efO2i). The shortest hydrogen-bond length is 3.1815\u2005(16)\u2005\u00c5 and is the strongest supra\u00admolecular inter\u00adaction with an angle of 162.8\u2005(17)\u00b0 (C17\u2014H17A\u22efO4). Analysis of the hydrogen-bonding network shows that all the hydrogen bonds shown in Table\u00a02D11(2); Etter et al., 1990The crystal packing along the \u03b1-amino\u00adsilane derivatives that are structurally based on compound 1 and its starting compound 2. Examples of such \u03b1-piperidino\u00adsilanes found in the Cambridge Structural Database -1-methyl-1-{[meth\u00adyl(phen\u00adyl)(tri\u00admethyl\u00adgerm\u00adyl)sil\u00adyl]meth\u00adyl}piperidinium iodide, C17H32GeNSiI meth\u00adyl]gall\u00adium n-pentane solvate, C45H67GaN2Si4\u00b70.5(C5H12) -5,6-aza-C60fulleroid, C79H17NSi (0.81\u2005mmol) was dissolved in acetone (3\u2005ml) and diethyl sulfate (0.81\u2005mmol) was added dropwise to the solution. The reaction mixture was stirred and heated for 6\u2005h at 329\u2005K. Afterwards the reaction was quenched by the addition of a mixture of H2O (2\u2005ml) and NH3 (2\u2005ml). The aqueous phase was extracted three times with CH2Cl2 and the combined organic phases were dried over Na2SO4. After the removal of volatile compounds, the raw product was dissolved in n-pentane (1\u2005ml) and stored at 243\u2005K. The title salt (1) was isolated as colorless crystalline blocks.The reaction scheme for the synthesis of 1H NMR : \u03b4 = 0.30 , 1.24\u20131.31 , 1.65\u20131.90 , 2.29 , 3.12 , 3.37\u20133.56 , 4.12 , 7.04 , 7.10\u20137.15 , 7.24 ppm.Uiso(H).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698902101361X/hb8003sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902101361X/hb8003Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902101361X/hb8003Isup3.cmlSupporting information file. DOI: 2131144CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I atom in the title compound, which exhibits strong blue emission, adopts a highly distorted trigonal\u2013planar geometry coordinated by two pyridine N atoms of two crystallographically independent 2\u2032,6\u2032-di\u00adfluoro-2,3\u2032-bi\u00adpyridine ligands and one O atom of the tri\u00adfluoro\u00admetane\u00adsulfonate anion.The Ag 3SO3)(C10H6F2N2)2], the AgI centre adopts a highly distorted trigonal\u2013planar coordination environment resulting from its coordination by one O atom of the tri\u00adfluoro\u00admethane\u00adsulfonate anion and the pyridine N atoms of two crystallographically independent 2\u2032,6\u2032-di\u00adfluoro-2,3\u2032-bi\u00adpyridine ligands, which display very similar conformations to one another. Pairwise Ag\u22efO\u2013SO2CF3\u2212 [Ag\u22efO = 2.8314\u2005(14)\u2005\u00c5] inter\u00adactions and inter\u00admolecular C\u2014H\u22efO inter\u00adactions between inversion-related units lead to the formation of an eight-membered cyclic dimer in which the silver atoms are separated by 6.2152\u2005(3)\u2005\u00c5. In the crystal, the dimers are linked through C\u2014H\u22efO hydrogen bonds, halogen\u22ef\u03c0 and weak \u03c0\u2013\u03c0 stacking inter\u00adactions, resulting in the formation of a three-dimensional supra\u00admolecular network. The title compound exhibits a strong and broad emission band from 400\u2005nm to 550\u2005nm in solution and its photoluminescence quantum efficiency is estimated to be ca 0.2, indicating that the title compound could have applications as an emitting material in organic light-emitting diodes (OLEDs).In the title compound, [Ag(CF III and PtII complexes by many researchers because of their applicability to OLEDs and solid-state lighting and 53.10\u2005(7)\u00b0, respectively. As shown in Fig.\u00a01I ion is coordinated by two pyridine N atoms (N2 and N4) from two 2\u2032,6\u2032-di\u00adfluoro-2,3\u2032-bi\u00adpyridine ligands and one O atom from the tri\u00adfluoro\u00admethane\u00adsulfonate anion, forming a highly distorted trigonal\u2013planar geometry. Selected bond lengths and angles around the Ag1 atom are given in Table\u00a012CF3\u2212 inter\u00adaction between the metal ion and an O atom of an adjacent tri\u00adfluoro\u00admethane\u00adsulfonate anion . The AgI atom is displaced out of the trigonal N2, N4, O1 coordination plane by 0.1057\u2005(9)\u2005\u00c5. The C6\u2013C10/N2 and C16\u2013C20/N4 pyridine rings coordinated to the AgI centre are tilted by 25.75\u2005(10)\u00b0 with respect to each other. The pairwise Ag\u22efO links lead to the formation of an eight-membered [Ag\u2014O\u2014S\u2014O\u2014]2 cyclic dimer, in which the silver atoms are separated by 6.2152\u2005(3)\u2005\u00c5. The cyclic dimer is consolidated by C\u2014H\u22efO inter\u00adactions (Table\u00a02The asymmetric unit in the title compound consists of an Ags Table\u00a02.Cg4\u22efCg4ii = 3.9737\u2005(11)\u2005\u00c5; Cg4 is the centroid of the C16\u2013C20/N4 ring; symmetry code: (ii) \u2212x, \u2212y, \u2212z\u00a0+\u00a01] between the pyridine rings, forming a chain structure propagating along the a-axis direction. Neighbouring chains are connected by halogen\u22ef\u03c0 inter\u00adactions , thereby generating a two-dimensional supra\u00admolecular network lying parallel to the ab plane. Finally, these networks are stacked along the c-axis direction and connected by halogen\u22ef\u03c0 and weak \u03c0\u2013\u03c0 stacking inter\u00adactions , resulting in the formation of a three-dimensional supra\u00admolecular network.In the extended structure, the dimers are linked through C19\u2014H19\u22efO3 hydrogen bonds Table\u00a02 and weak2Cl2 solution, the title compound exhibits a strong and broad emission band with \u03bbmax = 400\u2005nm, as shown in Fig.\u00a053SO3) unit, significant blue-shifted emissions (> 50\u2005nm) are observed as compared with bi\u00adpyridine based IrIII complexes . The observed emission of the title compound is therefore attributed to ligand-centered \u03c0\u2013\u03c0* transitions with a minor contribution of an Ag-based metal-to-ligand charge-transfer transition. Similar dual-emission behaviour has been noted for some AgI complexes with 2-methyl\u00adthio\u00adthia\u00adzole in MeOH (2\u2005ml) in the dark at room temperature and the mixture was stirred for 10\u2005min. After that, the mixture was slowly evaporated in the air and a dark environment to obtain crystals suitable for X-ray crystallographic analysis. 1H NMR \u03b4 8.67 , 8.62 , 7.88\u20137.80 , 7.37\u20137.34 , 7.0.8 . 19F NMR \u03b4 \u221269.7, \u221271.8, 79.1. Analysis calculated for C21H12F7N4O3SAg: C 39.33; H 1.89; N 8.74%; found: C 39.44, H 1.86, N 8.70%.All experiments were performed under a dry NUiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021011282/hb7990sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989021011282/hb7990Isup2.hklStructure factors: contains datablock(s) I. DOI: 2118061CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The salan ligands coordinate to the molybdenum center in a \u03ba2N,\u03ba2O fashion, forming a distorted octa\u00adhedral geometry. These complexes crystallized as di\u00admethyl\u00adformamide and methanol solvated species.Two cis-dioxomolybdenum complexes based on salan ligands with different backbones are reported. The first complex, dioxido{2,2\u2032-bis\u00ad(phenolato)}molybdenum(VI) di\u00admethyl\u00adformamide disolvate, [Mo(C20H18N2O2)O2]\u00b72C3H7NO , features a phenyl backbone, while the second complex, bis(aza\u00adnedi\u00adyl)]bis\u00ad(methyl\u00adene)}bis\u00ad)dioxidomolybdenum(VI) methanol disolvate, [Mo(C36H56N2O2)O2]\u00b72CH3OH , is based on a cyclo\u00adhexyl backbone. These complexes crystallized as solvated species, 1b\u00b72DMF and 2b\u00b72MeOH. The salan ligands PhLH2 (1a) and CyLH2 (2a) coordinate to the molybdenum center in these complexes 1b and 2b in a \u03ba2N,\u03ba2O fashion, forming a distorted octa\u00adhedral geometry. The Mo\u2014N and Mo\u2014O distances are 2.3475\u2005(16) and 1.9567\u2005(16)\u2005\u00c5, respectively, in 1b while the corresponding measurements are Mo\u2014N = 2.3412\u2005(12)\u2005\u00c5, and Mo\u2014O = 1.9428\u2005(10)\u2005\u00c5 for 2b. A key geometrical feature is that the N\u2014Mo\u2014N angle of 72.40\u2005(4)\u00b0 in CyLMoO2 is slightly less than that of the PhLMoO2 angle of 75.18\u2005(6)\u00b0, which is attributed to the flexibility of the cyclo\u00adhexane ring between the nitro\u00adgen as compared to the rigid phenyl ring in the PhLMoO2.Two In this system, the salan ligand PhLH2 (1a) coordinates to the molybdenum center in a \u03ba2N,\u03ba2O fashion, forming a distorted octa\u00adhedral geometry. The angles formed around the molybdenum core are 80.23\u2005(6)\u00b0 for O1\u2014Mo01\u2014N1, 157.78\u2005(6)\u00b0 for O1\u2014Mo01\u2014O2, 75.18\u2005(6)\u00b0 for N1\u2014Mo01\u2014N2, and 109.80\u2005(7)\u00b0 for O3\u2014Mo01\u2014O4. These angles are consistent with a system that is significantly distorted from octa\u00adhedral geometry with bond angles resulting from the salan ligand ranging from 75.18\u2005(6) to 84.38\u2005(7)\u00b0, while the angle between the \u2018oxo\u2019 oxygens of 109.80\u2005(7)\u00b0 is close to the ideal tetra\u00adhedral angle of 109.5\u00b0. Analogous bond angles in the second molecule in the unit cell are the same within 0.01\u2005\u00c5. The bond distances between the molybdenum center and ligand atoms for Mo01\u2014N1 and Mo01\u2014O1 are 2.3475\u2005(16) and 1.9567\u2005(16)\u2005\u00c5, respectively. The notable bond distances from the salan ligand are O1\u2014C1 at 1.377\u2005(2)\u2005\u00c5, N1\u2014C7 at 1.486\u2005(3)\u2005\u00c5, C2\u2014C7 at 1.515\u2005(3)\u2005\u00c5, N1\u2014C8 at 1.389\u2005(8)\u2005\u00c5, and C8\u2014C13 at 1.419\u2005(3)\u2005\u00c5. Analogous bond distances in the second molecule in the unit cell are the same within 0.01\u2005\u00c5 as distances for O1\u2014C1 and N1\u2014C8, respectively. The other bond distances have variations of 0.2\u20130.3\u2005\u00c5, with N3\u2014C27 at 1.519\u2005(3)\u2005\u00c5, C26\u2014C27 at 1.490\u2005(3)\u2005\u00c5, and C28\u2014C33 at 1.392\u2005(3)\u2005\u00c5.The asymmetric unit of CyLMoO2 (2b) contains one mol\u00adecule of CyLMoO2 and two mol\u00adecules of methanol (MeOH) binds in the same \u03ba2N,\u03ba2O fashion that complex 1b does. Fig.\u00a04CyLMoO2 with the hydrogen atoms removed for clarity. The complex also has a distorted octa\u00adhedral geometry with angles of O3\u2014Mo01\u2014O1 at 96.36\u2005(5)\u00b0, O1\u2014Mo01\u2014N1 at 76.73\u2005(4)\u00b0, N1\u2014Mo01\u2014N2 at 72.40\u2005(4)\u00b0, N2\u2014Mo01\u2014O2 at 78.91\u2005(4)\u00b0, O2\u2014Mo01\u2014O4 at 100.19\u2005(5)\u00b0, O2\u2014Mo01\u2014O3 at 94.58\u2005(5)\u00b0. These angles are between 5 and 10\u00b0 of the ideal 90\u00b0 for octa\u00adhedral geometry. The N1\u2014Mo01\u2014N2 angle at 72.40\u2005(4)\u00b0 is slightly less than that of the PhLMoO2 angle of 75.81\u2005(6)\u00b0, which is attributed to the flexibility of the cyclo\u00adhexane ring between the nitro\u00adgen atoms compared to the rigid phenyl ring in the PhLMoO2. Metal\u2013ligand bond distances are found for Mo01\u2014O1 at 1.9428\u2005(10)\u2005\u00c5, Mo01\u2014O2 at 1.9484\u2005(10)\u2005\u00c5, Mo01\u2014O3 at 1.7125\u2005(10)\u2005\u00c5, Mo01\u2014O4 at 1.7226\u2005(11)\u2005\u00c5, Mo01\u2014N1 at 2.3412\u2005(12)\u2005\u00c5, and Mo01\u2014N2 at 2.3384\u2005(12)\u2005\u00c5. Other ligand distances and bond lengths within the phenyl rings are consistent with analagous distances in PhLMoO2 (1b). The cylohexane bond distances are consistent with single C\u2014C bonds. The bond lengths observed are not statistically different than those reported by Ziegler et al. (2009R1 of 2.78% as compared to the previously reported R1 of 5.5% and higher solvent disorder in the reported structure.The asymmetric unit of H) Fig.\u00a03. The sal al. 2009. There aPhLMoO2 (1b): A single mol\u00adecule of PhLMoO2 is hydrogen bonded to one disordered DMF mol\u00adecule, as shown in Fig.\u00a05D with a distance of 2.16\u2005(3)\u2005\u00c5. Corresponding hydrogen bond distances in the second molecule in the unit cell are similar. There are three formula units within the contents of the unit cell. Perpendic\u00adular \u03c0-stacking between PhLMoO2 mol\u00adecules is observed between C5 and the aryl ring centroid (C35\u2013C39) with a distance of 4.597\u2005\u00c5.CyLMoO2 (2b): There are four mol\u00adecules of CyLMoO2 in the unit cell of this system and the complex is stabilized via hydrogen bonding to the solvent MeOH mol\u00adecule . There was a similar crystal structure found with the imine form of the ligand MoO2. A search for CyLMoO2 (2b) in the CSD shows that there is a known structure of the mol\u00adecule with a different unit cell with accession code HUWGOW was solved in space group P 21/n compared with P31 for HUWGOW. The primary additional differences in the structures is an improved R1 of 2.78% and more clearly resolved methanol solvent, as compared to the previously reported R1 of 5.5% and more disordered methanol solvent and CyLH2 (2a) were synthesized as off-white solids in 86% and 58% yields, respectively. The reaction scheme is shown in Fig.\u00a071H NMR spectra of both ligands as compared to the precursor salen compounds was the disappearance of the aldimine peak (\u223c8.50 ppm) and the appearance of the benzylic resonances \u223c4.00 ppm. The molybdenum complexes PhLMoO2 (1b) and CyLMoO2 (2b) were synthesized in 86% and 42% yields, respectively, by the reaction of the corresponding ligands with MoO2(acac)2 in methanol or aceto\u00adnitrile as solvent. Complexes 1b and 2b were also characterized by NMR and IR spectroscopy. Both complexes exhibited stretches { characteristic of a cis-dioxo molybdenum core in the IR spectrum.The salan ligands used for stabilizing molybdenum(VI) )dioxidomolybdenum(VI) : A round-bottom flask equipped with a magnetic stirring bar was charged with MoO2(acac)2 and methanol (ca. 10\u2005mL). The solution was stirred, and 2a was added to the MoO2(acac)2 dissolved in methanol. The solution was stirred overnight when it turned orange. The solution was filtered, and the solvent removed by evaporation under vacuum to obtain an orange precipitate. The precipitate was triturated with methanol, producing an orange solid, which was separated by gravity filtration and was washed twice with cold methanol . 1H NMR \u03b4 7.26 , 6.86 , 5.28 , 4.18 , 2.34\u20132.28 , 1.43 , 1.30 , 1.19\u20131.17 , 0.88\u20130.85 . 13C{1H} NMR \u03b4 157.1, 152.1, 142.8, 142.3, 142.0, 138.0, 137.7, 137.6, 125.7, 125.4, 124.1, 124.0, 123.0, 122.9, 120.0, 119.6, 65.19, 58.9, 57.6, 53.4, 50.9, 50.5, 35.2, 35.1, 34.3, 34.2, 33.0, 31.6, 31.6, 31.5, 29.9, 29.9, 28.9, 24.5, 24.3, 24.1. Selected IR (cm\u22121): 903, 875 \u03c5(Mo=O).bis\u00ad(aza\u00adnedi\u00adyl)]bis\u00ad(methyl\u00adene)}bis\u00ad = 1.2Ueq(C) for CH, CH2, and CH3. The structure for PhMoO2 (1b) was initially refined in the trigonal crystal system P3221; however, this resulted in the solvent DMF having a high level of disorder with many checkCIF errors.Crystal data, data collection, and refinement details are listed in Table\u00a0410.1107/S2056989022000524/tx2046sup1.cifCrystal structure: contains datablock(s) 2b, 1b. DOI: 10.1107/S2056989022000524/tx20462bsup3.hklStructure factors: contains datablock(s) 2b. DOI: 10.1107/S2056989022000524/tx20461bsup2.hklStructure factors: contains datablock(s) 1b. DOI: 2142074, 2142073CCDC references: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A three-dimensional network of hydrogen bonds serves to hold the structure together.The polymer contains Li 4(C3H2N3O3)4(H2O)7]n, synthesized at room temperature from an aqueous solution of lithium hydroxide and cyanuric chloride, crystallizes in the triclinic space group P+ cations in the asymmetric unit, one of which, Li1, has distorted trigonal\u2013bipyramidal geometry and is coordinated via oxygen to two cyanurate anions occupying equatorial positions, and three water mol\u00adecules, two in the axial positions and the third in an equatorial position. One of the axial water ligands and the equatorial water ligand are involved in bridging to a crystallographically equivalent Li1 cation. A centre of inversion lies between the two Li1 cations and the Li1\u22efLi1 distance is 3.037\u2005(5)\u2005\u00c5. The remaining axial water ligand bridges to the second Li cation, Li2, which is disordered over two crystallographic sites with approximately equal occupancy, and has an Li1\u22efLi2 distance of 3.438\u2005(7)\u2005\u00c5. The terminal Li2 cation is coordinated to three water mol\u00adecules and an oxygen atom from a cyanuric anion and has a distorted tetra\u00adhedral geometry. A three-dimensional network of inter\u00admolecular hydrogen bonds involving N\u2014H\u22efO, O\u2014H\u22efO and O\u2014H\u22efN inter\u00adactions serves to hold the structure together. The title compound was further characterized using IR and UV\u2013vis spectroscopy and TG\u2013DTA analysis.The polymeric title complex, poly[hexa-\u03bc-aqua-diaquatetra-\u03bc-cyanurato-tetralithium] [Li The complex has been characterized by single-crystal X-ray diffraction, FTIR and UV\u2013Vis spectroscopy, and TG\u2013DTA analysis.As cyanuric acid has three hydrogen-bonding-donor amine sites and three hydrogen-bonding-acceptor keto sites, it has been the subject of several structural and crystal-design studies 4(H2O)7] . This coordination preference may be due to the hard acid, Li+, preferring to bond to the harder base i.e. oxygen.The two crystallographically distinct cyanurate ligands exist in resonance form (IV) Fig.\u00a01, in whicet al., 2020A and Li2B) and have slightly longer bond lengths: C2\u2014O3, 1.2439\u2005(18)\u2005\u00c5; C3\u2014O2, 1.2436\u2005(19)\u2005\u00c5; C5\u2014O5, 1.2442\u2005(18)\u2005\u00c5 and C6\u2014O6, 1.2430\u2005(18)\u2005\u00c5. The delocalization of the negative charge on the deprotonated nitro\u00adgen atoms (N2 and N5) over the adjacent keto groups is shown as dashed lines in Fig.\u00a02The C=O groups involved in coordination to Li1, namely C1=O1 and C4=O4 have bond lengths of 1.2207\u2005(19) and 1.2242\u2005(19)\u2005\u00c5, respectively Fig.\u00a03, which a+ cations in the asymmetric unit in the axial positions and the third (H2O8) in an equatorial position. The Li1\u2014O bond lengths lie in the range 2.012\u2005(3)\u2013 2.201\u2005(3)\u2005\u00c5 and the bond angles of O4\u2014Li1\u2014O1 = 118.40\u2005(13)\u00b0, O4\u2014Li1\u2014O8 = 120.78\u2005(14)\u00b0, O1\u2014Li1\u2014O8 = 120.74\u2005(14)\u00b0 and O8i\u2014Li1\u2014O7 = 178.56\u2005(15)\u00b0 confirm the trigonal\u2013bipyramidal Li1 coordination geometry. One of the axial water ligands, H2O8i, and the equatorial water ligand, H2O7, bridge to a crystallographically equivalent Li1 cation. The Li1\u22efLi1i distance is 3.037\u2005(5)\u2005\u00c5, which is larger than the Li\u2014Li bond distance found in lithium metal. The Li1\u2014O\u2014Li1i bridge angle is 95.00\u2005(11)\u00b0. The Li1\u2014O8 and Li1\u2014O8i bond lengths are 2.032\u2005(3) and 2.086\u2005(3)\u2005\u00c5, respectively.There are two distinct Liit Fig.\u00a03. Li1 has2O7, bridges to the second Li+ cation, Li2, which is disordered over two sites, Li2A and Li2B, which have approximately equal occupancies. The Li1\u22efLi2A and Li1\u22efLi2B distances are 3.438\u2005(7) and 3.439\u2005(7)\u2005\u00c5, respectively. Li2 is coordinated to two more water mol\u00adecules, H2O9, H2O10 and an oxygen atom from a cyanurate ligand (either O3ii for Li2A or O2iii for Li2B) to complete its distorted tetra\u00adhedral coordination geometry. The Li2\u2014O bond lengths lie in the range 1.931\u2005(7)\u20132.057\u2005(7)\u2005\u00c5 and the O\u2014Li2\u2014O angles in the range 97.9\u2005(3)\u2013125.3\u2005(3)\u00b0The remaining axial water ligand, H4(C3H2N3O3)4(H2O)7] units into a three-dimensional network and 2.7443\u2005(16)\u2005\u00c5, respectively].Strong inter\u00admolecular hydrogen-bonding inter\u00adactions Table\u00a01 link therk Fig.\u00a04. These ii, N3\u2014H3\u22efO5ii, N4\u2014H4\u22efO6iii and N6\u2014H6\u22efO6iv). Weaker hydrogen-bonding inter\u00adactions, with N\u22efO distances in the range 2.905\u2005(2)\u20133.0342\u2005(19)\u2005\u00c5 are also observed between the unprotonated N atoms of the cyanurate ions and nearby water mol\u00adecules .In addition, each cyanurate moiety forms two strong hydrogen bonds between the N\u2014H groups and oxygen atoms of adjacent mol\u00adecules with N\u22efO distances in the range 2.7964\u2005(16)\u20132.8054\u2005(17)\u2005\u00c5 \u00adbis\u00addicopper(I) tetra\u00adaqua\u00adstrontium(II)] bis\u00ad(\u03bc-aqua)\u00adtetra\u00adaqua\u00adcopper(II)disodium(I)] and 3389\u2005cm\u22121 (br) correspond to \u03bd(O\u2014H) 1/2/h\u03bd, where t is the thickness of the crystal (1\u2005mm) and h\u03bd is the photon energy. A plot of (\u03b1h\u03bd)2versus h\u03bd is shown as the inset in Fig.\u00a07Eg) is estimated to be 5.22\u2005eV.The UV\u2013Vis NIR absorption spectrum was measured using a Perkin Elmer lambda 950 UV\u2013Vis\u2013NIR spectrophotometer Fig.\u00a06. The pea\u22121 with a resolution of 1\u2005\u00b5g under a dry N2 atmosphere. The thermogram .Simultaneous TG\u2013DTA measurements and analysis of weight change and heat flow were performed using a Perkin Elmer STA 6000 instrument operating at a scanning rate of 10\u00b0C minam Fig.\u00a07 shows foLithium hydroxide and cyanuric chloride were dissolved in water (100\u2005ml). The resulting solution was stirred for 5\u2005h at ambient temperature (300-301\u2005K) and filtered twice using Whatman filter paper. The solvent was allowed to evaporate in a dust-free environment. After 22 days, good quality colourless crystals were harvested.A and Li2B, which were resolved using the PART command = 1.2Ueq(N). The hydrogen atoms on the water mol\u00adecules were located in difference-Fourier maps and each Uiso(H) parameter was freely refined with the O\u2014H distance restrained to 0.85\u2005(2)\u2005\u00c5 using DFIX. The H\u2014O\u2014H angle distances were restrained using DFIX to a target value of 1.39\u2005(2)\u2005\u00c5 [or 1.41\u2005(2)\u2005\u00c5 for H9A\u2014O9\u2014H9B] in order to keep the water mol\u00adecules close to their standard geometries.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021009324/cq2044sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021009324/cq2044Isup2.hklStructure factors: contains datablock(s) I. DOI: 1991191CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "To evaluate the effect of toric intraocular lens implantation in cataract patients with irregular corneal steep and flat meridian. Data of 112 eyes of 78 patients who underwent toric intraocular lens implantation were analyzed retrospectively. Steep meridian deviations (not 180\u00b0) and steep and flat meridian deviations (not 90\u00b0) were classified as 0, 1\u20139, 10\u201319, 20\u201329, 30\u201339, and over 30\u00b0. Meridian deviation was measured with a sagittal map of a rotating Scheimpflug camera using PicPickTools . D) of 0 and 1\u20139 groups were significantly lower than that of 10\u201319 , 20\u201329 , and over 30\u00b0 groups both in steep meridian deviations and horizontal and vertical meridian deviations at 6 months (P < 0.05). Postoperative mean UCVA (logMAR) of 0 (logMAR) and 1\u20139 groups was significantly improved compared to that of 10\u201319 , 20\u201329 , and over 30\u00b0 groups both in steep meridian deviations and horizontal and vertical meridian deviations at 6 months (P < 0.05). Residual astigmatism ( Correction of astigmatism with toric intraocular lens implantation is not accurate in corneas with steep meridian deviations and steep and flat meridian deviations of more than 10\u00b0. Therefore, care should be taken when we perform toric intraocular lens implantation in patients with irregular corneal meridian. D) of corneal astigmatism [D or greater [Cataract is the most common cause of visual impairment in elderly people . About 6igmatism , and 20% greater . If this greater . Toric i greater . Toric I greater . TherefoTo the best of our knowledge, no studies have reported the efficacy of toric IOL implantation in normal patients with irregular corneal astigmatism such as steep meridian deviations or steep and flat meridian deviation. Thus, the objective of this study was to evaluate outcomes of toric IOL implantation according to the amount of steep meridian deviations and angle deviation of steep and flat meridian.We performed a retrospective chart review and data analysis in this study. This study was conducted in compliance with Institutional Review Board regulations, informed consent regulations, sponsor and investigator obligations, and the Declaration of Helsinki. The Institutional Review Board (IRB)/Ethics Committee of Bucheon St. Mary Hospital approved this study protocol.A total of 112 eyes of 78 patients who underwent toric intraocular lens implantation in Bucheon St. Mary Hospital from July 2017 to April 2018 were enrolled. Steep meridian deviations (not 180\u00b0) were classified as 0, 1\u20139, 10\u201319, 20\u201329, 30\u201339, and over 30\u00b0. Steep and flat meridian deviations (not 90\u00b0) were classified as 0, 1\u20139, 10\u201319, 20\u201329, and over 30\u00b0. Inclusion criteria were advanced cataracts in patients with high corneal astigmatism more than 1.5D. Exclusion criteria were a history of any ocular injury or disorder, infection, inflammation, and surgery within prior 6 months.Steep meridian deviations and steep and flat meridian deviation were measured with a sagittal map of a rotating Scheimpflug camera using PicPickTools . Steep meridian deviation was determined as an absolute value of 180 minus the steep axis angle. Steep and flat meridian deviation was determined as an absolute value of 90 minus steep and flat meridian angle .\u00ae, Oculus, Germany).All patients underwent a complete ophthalmological examination. Their demographic and perioperative data were recorded. Uncorrected and corrected distance visual acuities were expressed as logMAR. Manifest refraction, biometry, and keratometry with the IOLMaster partial coherence interferometry device (Carl Zeiss Meditec AG), corneal topography , slit lamp examination, and dilated funduscopy were examined at the preoperative period and postoperative 2, 4, and 6 months. The IOL manufacturer's web-based toric calculator was used to determine the required cylinder power and axis for the IOL that was going to be implanted. The total corneal astigmatism was measured using the Scheimpflug system using an Intrepid Infiniti system . Corneal steep axis and 6.0\u2009mm ring were marked with gentian violet. Surgery was performed through a clear corneal incision at the steep astigmatic axis. After topical ocular anesthesia was applied, a 2.75\u2009mm clear corneal incision was made using a 2.75\u2009mm double-blade keratome (Alcon). Surgically induced astigmatism was set by 0.5 diopters. Sodium hyaluronate 1.0% was used to reform and stabilize the anterior chamber. A continuous curvilinear capsulotomy was made according to a 6.0\u2009mm corneal marker using Inamura capsulorhexis forceps . Hydrodissection and hydrodelineation were achieved using a balanced salt solution. Phacoemulsification was performed using 2.75\u2009mm-sized phacotips and infusion/aspiration (I/A) cannulas for micro- and small-incision groups, respectively. A clear preloaded IOL was implanted in the capsular bag. The IOL was rotated to the correct axis position according to the axis of total corneal astigmatism. The wound was not sutured. Postoperative treatment consisted of gatifloxacin 0.3% and fluorometholone acetate 0.01% eye drops four times a day for four weeks.P values\u2009<\u20090.05 were considered statistically significant.All statistical analyses were performed using a commercial program . The Wilcoxon signed rank test was used to compare pre- and postoperative BCVA and refractive and keratometer astigmatisms. A total of 112 eyes of 78 patients were enrolled in this study. Preoperative mean autorefractive cylinder was 2.21\u2009\u00b1\u20091.36 D. Mean total corneal astigmatism measured with the Scheimpflug camera was 2.35\u2009\u00b1\u20091.15 D. Mean UCVA was 0.83\u2009\u00b1\u20090.35 (logMAR), and mean BCVA was 0.56\u2009\u00b1\u20090.29 (logMAR) .P < 0.05). Postoperative mean total corneal astigmatism (1.96\u2009\u00b1\u20091.0 D) was also decreased compared to preoperative value (2.35\u2009\u00b1\u20091.15 D), but the decrease was statistically insignificant. Mean IOL rotation was 3.2\u2009\u00b1\u20091.5\u00b0 at 2 months after toric IOL implantation. Postoperative mean UCVA (0.09\u2009\u00b1\u20090.06) (logMAR) and mean BCVA (0.02\u2009\u00b1\u20090.01) were improved compared to preoperative values (both P < 0.05) was significantly decreased compared to its preoperative value (2.21\u2009\u00b1\u20091.36 D) ( < 0.05) .D) and mean UCVA (logMAR) of all groups were similar to each other . Postoperative mean UCVA (logMAR) of 0 (0.09\u2009\u00b1\u20090.04) (logMAR) and 1\u20139\u00b0 (0.10\u2009\u00b1\u20090.04) groups was significantly improved compared to that of 10\u201319 (0.14\u2009\u00b1\u20090.05), 20\u201329 (0.18\u2009\u00b1\u20090.08), and over 30\u00b0 groups (0.20\u2009\u00b1\u20090.09) in steep meridian deviations at 6 months (D) values of 0 (0.51\u2009\u00b1\u20090.13) and 1\u20139\u00b0 groups (0.61\u2009\u00b1\u20090.16) were significantly lower than those of 10\u201319 (0.92\u2009\u00b1\u20090.24), 20\u201329 (0.10\u2009\u00b1\u20090.32), and over 30\u00b0 groups (1.12\u2009\u00b1\u20090.37) in steep meridian deviations at postoperative 6 months (P < 0.05) (D) . Estimat < 0.05) . There w < 0.05) .P > 0.05). However, preoperative total corneal astigmatism (D) of 21\u201330\u00b0 (2.04\u2009\u00b1\u20090.86 D) and over 30\u00b0 groups (2.06\u2009\u00b1\u20091.05 D) was statistically higher than that of other groups (P < 0.05). Postoperative mean UCVA (logMAR) values of 0 (0.09\u2009\u00b1\u20090.05) and 1\u20139\u00b0 (0.11\u2009\u00b1\u20090.08) were significantly improved compared to those of 10\u201319 (0.17\u2009\u00b1\u20090.10), 20\u201329 (0.21\u2009\u00b1\u20090.10), and over 30\u00b0 groups (0.22\u2009\u00b1\u20090.11) in steep and flat meridian deviations at 6 months (P < 0.05) (D) of 0 (0.55\u2009\u00b1\u20090.15) and 1\u20139\u00b0 groups (0.66\u2009\u00b1\u20090.19) was significantly lower than that of 10\u201319 (0.90\u2009\u00b1\u20090.28), 20\u201329 (1.01\u2009\u00b1\u20090.35), and over 30\u00b0 groups (1.14\u2009\u00b1\u20090.40) in steep and flat meridian deviations at postoperative 6 months (P < 0.05) (D) values of all groups were similar to each other ( < 0.05) . Estimat < 0.05) . There w < 0.05) .D depending on preoperative astigmatism [Implantation of toric IOL in cataract surgery to correct corneal astigmatism has been popular due to its excellent clinical outcomes and increased patient demands . Howeverigmatism . Severaligmatism , ignorinigmatism , inaccurigmatism . Zhu et igmatism . In the If there are severe steep meridian deviations, the other marker of toric IOL and corneal steep meridian may be misaligned despite that surgeons have made a perfect alignment of one marker of toric IOL and corneal steep meridian. If there is a severe steep and flat meridian deviation, the toric IOL steep axis and corneal flat axis may be misaligned despite that the surgeon has made a perfect alignment of marker of toric IOL and corneal steep meridian.D) values of 0 (0.51\u2009\u00b1\u20090.13) and 1\u20139\u00b0 groups (0.61\u2009\u00b1\u20090.16) were significantly lower than those of 10\u201319 (0.92\u2009\u00b1\u20090.24), 20\u201329 (0.10\u2009\u00b1\u20090.32), and over 30\u00b0 groups (1.12\u2009\u00b1\u20090.37) in steep meridian deviations at postoperative 6 months (P < 0.05) (D) and mean UCVA (logMAR) values of all groups were similar to each other (P > 0.05). However, postoperative mean UCVA (logMAR) of 0 (0.09\u2009\u00b1\u20090.04) (logMAR) and 1\u20139\u00b0 (0.10\u2009\u00b1\u20090.04) groups was significantly improved compared to that of 10\u201319 (0.14\u2009\u00b1\u20090.05), 20\u201329 (0.18\u2009\u00b1\u20090.08), and over 30\u00b0 groups (0.20\u2009\u00b1\u20090.09) in steep meridian deviation at 6 months (P < 0.05) . Preoper < 0.05) .D) values of 21\u201330\u00b0 and over 30\u00b0 groups were statistically higher than those of other groups (P < 0.05) . We hypoD) of 0 (0.55\u2009\u00b1\u20090.15) and 1\u20139\u00b0 groups (0.66\u2009\u00b1\u20090.19) were significantly lower than that of 10\u201319 (0.90\u2009\u00b1\u20090.28), 20\u201329 (1.01\u2009\u00b1\u20090.35), and over 30\u00b0 groups (1.14\u2009\u00b1\u20090.40) in steep and flat meridian deviations at postoperative 6 months (P < 0.05) (P < 0.05) . Also, p < 0.05) .D) at 6 months (D) . A posit < 0.05) was alsoP < 0.05). Therefore, patients who had meridian deviations of less than 10\u00b0 had significantly lower residual astigmatism and better uncorrected visual acuity than those who had meridian deviation of more than 10\u00b0.Toric IOL rotations of less than 10\u00b0 changed the eye's refractive astigmatism to less than 0.50 diopters . In the D) also increased in patients with irregular corneal astigmatism. Therefore, the effect of toric IOL implantation is optimized in patients with regular corneal astigmatism. Toric IOL implantation should be performed cautiously when patients have steep meridian deviations or steep and flat meridian deviations.To the best of our knowledge, this was the first study to evaluate the outcomes of toric IOL implantation in cataract patients with irregular corneal astigmatism. The postoperative visual acuity of patients with regular astigmatism was significantly improved compared to that of patients with steep meridian deviations and the steep and flat meridian deviations. If steep meridian deviations and steep and flat meridian deviations increased, estimated residual astigmatism (A multicenter clinical trial with a larger sample size and longer follow-up period is needed to observe the long-term efficacy of toric intraocular lens implantation in cataract patients with irregular corneal astigmatism."} +{"text": "The packing of the units is dominated by C\u2014H\u22efO hydrogen bonding and weak dispersion forces, with a minor contribution from C\u2014H\u22ef\u03c0 bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions.The structure of tel\u00adlur\u00adium(VI)], [Sb3Te(C6H5)9O6], the hexa\u00adoxidotellurate(VI) ion is coordinated to three SbV ions via pairs of cis-positioned O atoms to form a discrete mol\u00adecular unit. The TeVI and SbV central ions exhibit distorted octa\u00adhedral [TeO6] and distorted trigonal\u2013bipyramidal [SbC3O2] coordination geometries, respectively. The linking of these polyhedra, by sharing the dioxide edges, results in the Te-based octa\u00adhedron having a mer-configuration. The packing of the mol\u00adecules is dominated by C\u2014H\u22efO hydrogen bonding and weak dispersion forces, with a minor contribution from C\u2014H\u22ef\u03c0 bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions. According to the Hirshfeld surface analysis, the contributions of the H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH contacts are 58.0, 32.6 and 7.8%, respectively. The title structure provides a model for the bonding of triorgano\u00adanti\u00admony dications to octa\u00adhedral oxoanions, and the observed doubly bridged motifs, Te(\u03bc-O)2Sb, may find application in the functionalization of polyoxometalate species.In the structure of the title compound [systematic name hexa-\u03bc-oxido-1:2\u03ba The latter are formally similar to 1,2-benzene\u00addiolate chelates, which have been observed in mol\u00adecular organo\u00adanti\u00admony compounds 3TeO6, which features the formation of discrete clusters, [Te{(\u03bc-O)2SbPh3}3].At the same time, PhC2/c, and contains the discrete mol\u00adecular unit shown in Fig.\u00a016] octa\u00adhedron and three [Ph3SbO2] polyhedra sharing oxide edges. Thus two oxide bridges are formed from TeVI to each of the three SbV ions with Te\u2014O\u2014Sb angles in the range 99.33\u2005(13)\u2013102.41\u2005(13)\u00b0 \u2005\u00c5, which occurs in the Te1(\u03bc-O)2Sb2 unit. Such fully substituted organometallic hexa\u00adoxotellurate(VI) units are exceedingly rare, with the only known example being an aliphatic SnIV derivative 4Te(OH)2] and [(Ph3SnO)2Te(OMe)4] \u2013174.49\u2005(13)\u00b0. The fivefold coordination around each of three Sb-atoms can best be described as distorted trigonal bipyramidal, with the O2\u2014Sb1\u2014C7 = 161.09\u2005(15)\u00b0, O4\u2014Sb2\u2014C19 = 164.73\u2005(16)\u00b0 and O5\u2014Sb3\u2014C37 = 165.43\u2005(16)\u00b0 bond angles defining the principal axes of the trigonal bipyramids. This assignment is supported by the calculated five-coordinate \u03c4-indices, which are 0.69, 0.75 and 0.65 for Sb1, Sb2 and Sb3, respectively \u20132.110\u2005(3)\u2005\u00c5] than the equatorial Sb\u2014Oeq bonds, Sb1\u2014O1, Sb2\u2014O3 and Sb3\u2014O6 [in the range 1.966\u2005(3)\u20131.992\u2005(3)\u2005\u00c5]. This observation coincides with the differentiation of the Te\u2014O bond lengths; three of which, Te1\u2014O2, Te1\u2014O4 and Te1\u2014O5, lie in the range 1.904\u2005(3)\u20131.918\u2005(3)\u2005\u00c5 and three, Te1\u2014O1, Te1\u2014O3 and Te1\u2014O6, lie in the range 1.949\u2005(3)\u20131.968\u2005(3)\u2005\u00c5. Thus when considering the six Te\u2014O\u2014Sb bridges, the shorter Sb\u2014O bonds are accompanied by the longer Te\u2014O bonds and vice versa. The distribution of the Te\u2014OaxSb and Te\u2014OeqSb bonds indicates that the coordination octa\u00adhedron around the Te atom has the mer-configuration 3Te(OH)3] \u2005\u00c5; symmetry code (ii) x, y\u00a0+\u00a01, z; Table\u00a02b direction iii and C41\u2014H41\u22efCg(C13\u2013C18)iv x, \u2212y\u00a0+\u00a02, x\u00a0\u2212\u00a0x, \u2212y\u00a0+\u00a01, z\u00a0+\u00a0bc plane. In addition, to further consolidate the bilayers, there are weak slipped \u03c0\u2013\u03c0 stacking inter\u00adactions between pairs of inversion-related phenyl rings, with a centroid-to-centroid distance, Cg(C1\u2013C6)\u22efCg(C1\u2013C6)v = 3.807\u2005(6)\u2005\u00c5, an inter\u00adplanar distance of 3.603\u2005(5)\u2005\u00c5 and a slippage angle of 18.8\u2005(5)\u00b0 . There are no specific inter\u00adactions between the bilayers, and the shortest of their C\u22efC contacts [3.404\u2005(6)\u2005\u00c5] is not accompanied by any \u03c0\u2013\u03c0 overlap.Very weak mutual C\u2014H\u22efO bonding and trans-[(Ph3SnO)2Te(OMe)4] 2[CH2(Ph2SnO)2]2Te -octa\u00adkis\u00ad(tri\u00admethyl\u00adsil\u00adyloxy)ditellurium and orthotelluric acid tris\u00adester 3Te(OH)3, 4Te(\u03bc-O)2Te(OR)4 . The latter contains double Te\u2014O\u2014Te bridges, which are formally similar to the double Te\u2014O\u2014Sb bridges found in the title compound. No tetra\u00adhedral TeO4 fragments have been reported in organometallic series to date. The only known example of a tetra\u00adhedral tellurate is the ionic salt [NEt4]2TeO4\u00b72H2O . These results are consistent with the weakness of the C\u2014H\u22efO bonds in the structure. It is evident that only a few of the H\u22efC/C\u22efH contacts correspond to C\u2014H\u22ef\u03c0 bonding. Therefore, the H\u22efC/C\u22efH plot represents a rather diffuse collection of points between the pair of poorly resolved features and there no \u2018wings\u2019 at the upper left and lower right, which are characteristic of C\u2014H\u22ef\u03c0 inter\u00adactions bromide as follows:In previously reported syntheses, a range of silver salts were used in ion-exchange reactions to form Ph3H3TeO6, was synthesized according to the method of Gospodinov (19923H3TeO6 were added to a solution containing 0.612\u2005g (1.2\u2005mmol) of Ph4SbBr in 20\u2005mL of aceto\u00adnitrile. The mixture was stirred for 3\u2005h and then the AgBr precipitate removed by filtration. Evaporation of the solution yielded a colourless glassy material, which was then dissolved in 10\u2005mL of a 1:1 v/v mixture of benzene and butyl acetate. Slow evaporation of the solution to a volume of 2\u20133\u2005mL afforded 0.138\u2005g (27%) of the product in the form of long colourless prisms. The crystals were filtered and dried in air. Analysis (%) for C54H45O6Sb3Te: Found: C 50.12, H 3.39; Calculated: C 50.56, H 3.54. IR : 454s, 520m, 610s, 692vs, 732vs, 772w, 996w, 1066m, 1434s, 1478m, 1576w, 2824w, 3052m.The starting material, Aginov 1992. 0.220\u2005gUiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021011294/cq2048sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989021011294/cq2048Isup2.hklStructure factors: contains datablock(s) I. DOI: 2118082CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of 2-(2-hy\u00addroxy\u00adphen\u00adyl)quinoline-6-sulfonamide contains two crystallographically independent mol\u00adecules. The crystal structure features hydrogen bonding and \u03c0\u2013\u03c0 stacking inter\u00adactions. 15H12N2O3S, there are two mol\u00adecules (A and B) in the asymmetric unit. The attached phenol and quinoline moieties of each mol\u00adecule are almost coplanar with a dihedral angle of 6.05\u2005(15)\u00b0 for mol\u00adecule A and 1.89\u2005(13)\u00b0 for mol\u00adecule B. The crystal structure features N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions. Hirshfeld surface analysis indicates that the most significant contacts in the crystal packing are C\u22efH/H\u22efC (29.2%), O\u22efH/H\u22efO (28.6%) and H\u22efH (28.5%).In the title compound, C This compound was prepared in a two-step reaction, A and B). The C6A\u2014C7A and C6B\u2014C7B bond lengths of 1.472\u2005(5) and 1.470\u2005(5)\u2005\u00c5, respectively, are notably shorter than the normal C\u2014C single bond due to conjugation but are comparable to those observed in related structures ortho-position of each independent mol\u00adecule in (I)S(6) ring motifs , with a dihedral angle between their planes of 4.0\u2005(2)\u00b0 for mol\u00adecule A and 1.49\u2005(17)\u00b0 for mol\u00adecule B.The hydroxyl group in the fs Fig.\u00a01, which sA and 1.89\u2005(13)\u00b0 for mol\u00adecule B , indicating a significant electron delocalization within the mol\u00adecules. The sulfonamide groups are twisted away from the attached quinoline fragment with an C11A\u2014C12A\u2014S1A\u2014N2A torsion angle of 91.8\u2005(4)\u00b0 for mol\u00adecule A and C11B\u2014C12B\u2014S1B\u2014N2B torsion angle of \u2212 79.9\u2005(3)\u00b0 for mol\u00adecule B. The sulfonamide atoms S1A and S1B deviate by 0.228\u2005(1) and 0.054\u2005(1)\u2005\u00c5 from the planes of the quinoline fragment in mol\u00adecules A and B respectively.The attached quinoline and phenol moieties are almost coplanar with a dihedral angle of 6.05\u2005(15)\u00b0 for mol\u00adecule B Fig.\u00a02b, indica). The packing diagram of the title compound viewed down the a axis shows that the layers are stacked perpendicular to the b axis at and . These layers are formed by aggregation of c). In addition, the hydroxyl group of each mol\u00adecule is involved in a C\u2014H\u22efO hydrogen bond, forming an inversion dimer with an d). Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions are also observed in the crystal packing, forming a chain along the a-axis direction .In the crystal of (I)is Fig.\u00a03b shows fs Fig.\u00a03c. In adup Fig.\u00a03d. Weak on Fig.\u00a03e.Cg2\u22efCg3 = 3.779\u2005(2)\u2005\u00c5 for A mol\u00adecules and Cg6\u22efCg7 = 3.6636\u2005(18)\u2005\u00c5 for B mol\u00adecules where Cg2, Cg3, Cg6 and Cg7 are the centroids of the C1A\u2013C6A, C10A\u2013C15A, C1B\u2013C6B and C10B\u2013C15B rings, respectively]. These result in the formation of a supra\u00admolecular ribbon parallel to the a axis based on the stacked mol\u00adecules .Cohesion of the crystal structure is enhanced by the presence of \u03c0\u2013\u03c0 stacking inter\u00adactions, the most significant being between the 2-hy\u00addroxy\u00adphenyl and benzene rings of the quinoline groups of each mol\u00adecule .Crystal data, details of data collection, and results of structure refinement are summarized in Table\u00a0310.1107/S2056989022002870/zn2017sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022002870/zn2017Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022002870/zn2017Isup3.cmlSupporting information file. DOI: 2158517CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Apart from a slight site occupancy difference for the water mol\u00adecule of crystallization, compounds (I) and (II) are isostructural, featuring isolated tetra\u00adhedral cations of copper(I) ions coordinated by two dmbpy ligands and centrosymmetric, octa\u00adhedral anions of fluorinated early transition metals.The syntheses and crystal structures of two bimetallic mol\u00adecular compounds, namely, bis\u00ad\u00adcopper(I) hexa\u00adfluorido\u00adzirconate(IV) 1.134-hydrate, [Cu(dmbpy) 2]2[ZrF6]\u00b71.134H2O , (I), and bis[bis\u00ad\u00adcopper(I)] hexa\u00adfluorido\u00adhafnate(IV) 0.671-hydrate, [Cu(dmbpy)2]2[HfF6]\u00b70.671H2O, (II), are reported. Apart from a slight site occupany difference for the water mol\u00adecule of crystallization, compounds (I) and (II) are isostructural, featuring isolated tetra\u00adhedral cations of copper(I) ions coordinated by two dmbpy ligands and centrosymmetric, octa\u00adhedral anions of fluorinated early transition metals. The tetra\u00adhedral environments of the copper complexes are distorted owing to the steric effects of the dmbpy ligands. The extended structures are built up through Coulombic inter\u00adactions between cations and anions and \u03c0\u2013\u03c0 stacking inter\u00adactions between heterochiral \u0394- and \u039b-[Cu(dmbpy)2]+ complexes. A comparison between the title compounds and other [Cu(dmbpy)2]+ compounds with monovalent and bivalent anions reveals a significant influence of the cation-to-anion ratio on the resulting crystal packing architectures, providing insights for future crystal design of distorted tetra\u00adhedral copper compounds.The syntheses and crystal structures of two bimetallic mol\u00adecular compounds, namely, bis[bis\u00ad\u00adcopper(I)] hexa\u00adfluorido\u00adzir\u00adcon\u00adate(IV) 1.134-hydrate, [Cu(dmbpy) The octa\u00adhedral coordination environment of Zr1 is slightly distorted, with Zr1\u2014F bond lengths ranging from 1.9955\u2005(13) to 2.0183\u2005(12)\u2005\u00c5 Fig.\u00a01. The strd Table\u00a01. To qu\u00ada\u00c5 Table\u00a01. The min2 Table\u00a02].2]2[HfF6]\u00b70.671H2O and crystallizes in the triclinic space group P2]+ cations also have C2 symmetry, with the angle between the least squares planes containing Cu1 and each ligand being 84.14\u2005(8)\u00b0 \u00b0 between the N1/C1\u2013C5 and N2/C6\u2013C10 rings and 7.97\u2005(8)\u00b0 between the N3/C13\u2013C17 and N4/C18\u2013C22 rings. Moreover, the octa\u00adhedral coordination environment of Hf1 is also slightly distorted, with Hf1\u2014F bond lengths ranging from 1.9945\u2005(10) to 2.0111\u2005(11)\u2005\u00c5. Like in compound (I)trans position of HfF62\u2212 anions, but the geometry of the hydrogen bond is slightly different from that in compound (I)B\u22efF2 (Table\u00a04Compound (II) Fig.\u00a02. Compoun\u00b0 Table\u00a03 and the 2 Table\u00a04].2]+ cations and octa\u00adhedral MF62\u2212 anions are closely packed via Coulombic inter\u00adactions of 3.6967\u2005(12) and 3.7016\u2005(8)\u2005\u00c5, for compounds (I)2]+ pairs pack into racemic chains along the c-axis direction with heterochiral parallel displaced \u03c0\u2013\u03c0 inter\u00adactions between the N3/C13\u2013C17 and N4/C18\u2013C22 rings with an inter\u00adplanar angle of 0\u00b0, inter\u00adplanar distances of 3.708 and 3.678\u2005\u00c5, and centroid\u2013centroid distances (dpy\u2013py) of 5.3726\u2005(13) and 5.3777\u2005(11)\u2005\u00c5, for compounds (I)MF62\u2212 anions with hydrogen-bonded water mol\u00adecules are inter\u00adlaced between the racemic chains to form the extended three-dimensional structure. Compared to other mol\u00adecular compounds with MF62\u2212 anions in an extended and complicated hydrogen network In the extended structures of compounds (I)ns Fig.\u00a03. The \u0394/\u039bet al., 20162]+ complexes: [Cu(dmbpy)2][BF\u00ad4] et al., 2015A survey of compounds related to compounds (I)MF62\u2212, the compounds reported in the CSD are charge-balanced by monovalent anions and display two different types of packing architectures distinct from those of the title compounds: [Cu(dmbpy)2][BF\u00ad4], [Cu(dmbpy)2][PF\u00ad6], and [Cu(dmbpy)2][ClO\u00ad4] are isostructural, crystallizing in space group 1/cP2. Compared to compounds (I)Unlike compound (I)2][C16H9O8]\u00b7H2O, which crystallizes in space group P2]+ cations, \u03c0\u2013\u03c0 inter\u00adactions in the compound [Cu(dmbpy)2][C16H9O8]\u00b7H2O are dominant between [Cu(dmbpy)2]+ cations and [C16H9O8]\u2212 anions instead of between [Cu(dmbpy)2]+ cations. In this compound, the [Cu(dmbpy)2]+ cations and [C16H9O8]\u2212 anions are packed into charge-neutral chains via Coulombic inter\u00adactions and \u03c0\u2013\u03c0 inter\u00adactions along c axis and inversion centers are present between the chains. Additionally, the [C16H9O8]\u2212 anions and free water mol\u00adecules generate a three-dimensional network via O\u2014H\u22efO hydrogen bonding inter\u00adactions, resulting in a different architecture.Another type of packing architecture is found in [Cu(dmbpy)et al., 1993\u22121 to 423\u2005K and held at 423\u2005K for 24\u2005h. The autoclaves were allowed to cool to room temperature at a rate of 6\u2005K\u2005h\u22121. Orangish red solid products were recovered by vacuum filtration with a moderate yield. Compound (I)2, 0.835\u2005mmol of 6,6\u2032-dimethyl-2,2\u2032-bipyridyl, 0.15\u2005ml (4.14\u2005mmol) of HF (aq) (48%), and 0.1\u2005ml (5.5\u2005mmol) of deionized H2O. Compound (II)2, 0.835\u2005mmol of 6,6\u2032-dimethyl-2,2\u2032-bipyridyl, 0.05\u2005ml (1.38\u2005mmol) of HF (aq) (48%), and 0.2\u2005ml (11\u2005mmol) of deionized H2O.The compounds reported here were synthesized by the hydro\u00adthermal pouch method (Harrison Uiso(H) = 1.2Ueq(C) within OLEX2 I, II. DOI: 10.1107/S2056989021007295/hb7977Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989021007295/hb7977IIsup3.hklStructure factors: contains datablock(s) II. DOI: 2096336, 2096335CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Both the organic cation and the quasi-regular octa\u00adhedral inorganic anion are located about inversion centres. The organic cations and [SnCl6]2\u2212 anions lie in layers parallel to the ac plane with p-anisaldehyde mol\u00adecules occupying the space between the layers. A network of classical N\u2014H\u22efCl and N\u2014H\u22efO hydrogen bonds exists between the ethyl\u00adenedi\u00adammonium cations and the [SnCl6]2\u2212 anions and p-anisaldehyde mol\u00adecules. These inter\u00adactions, together with non-classical C\u2014H\u22efO inter\u00adactions between the ethyl\u00adenedi\u00adammonium cations and the p-anisaldehyde mol\u00adecules, serve to hold the structure together. The crystal studied was refined as a two-component twin.The asymmetric unit of the title organic\u2013inorganic hybrid complex , (C The Sn\u2014Cl2 bond involved in hydrogen bonding is slightly longer, at 2.4322\u2005(11)\u2005\u00c5, than the other Sn\u2014Cl bonds [Sn\u2014Cl1 = 2.4100\u2005(12)\u00c5 and Sn\u2014Cl3 = 2.4220\u2005(11)\u2005\u00c5]. These results are comparable to those reported by other research groups dianion, [SnClde Fig.\u00a01. The envac plane in which each [SnCl6]2\u2212 dianion is surrounded by four ethyl\u00adenedi\u00adammonium cations \u2005\u00c5 for N1\u22efO2iii to 3.404\u2005(4)\u2005\u00c5 for N1\u22efCl3v. Non-classical inter\u00adactions between the p-anisaldehyde mol\u00adecules and the ethyl\u00adenedi\u00adammonium cations, C9\u2014H9\u22efO2vi at 2.62\u2005\u00c5, further serve to hold the structure together.The packed crystal structure contains sheets lying parallel to the ns Fig.\u00a02. The p-ats Fig.\u00a03. The cryts Fig.\u00a03. The NH36H22N4)[SnCl6]Cl2\u00b72H2O and (C8H24N4)[SnCl6]Cl2\u00b72H2O [SnCl6] 2[SnCl6], and (C7H10N)2[SnCl6] 3SnBr6\u00b7Br ] were purchased from Sigma-Aldrich and were used without any further purification. The solvent use for the synthesis was ethanol (96%).Chemicals \u00b72C8H8O2 crystallizes in the space group P21/n with the monoclinic angle, \u03b2, close to 90\u00b0. The crystals formed as non-merohedral twins with about one quarter of reflections overlapping. The twin law corresponds to rotation about c*. For the crystal investigated, the relative domain sizes amounted to 0.790\u2005(4): 0.210\u2005(4). The structure was solved by intrinsic phasing = 1.2Uiso(C) or 1.5Uiso(C-meth\u00adyl)]. H atoms attached to N were located as local maxima in a difference-Fourier map and refined with a distance restraint N\u2014H = 0.9\u2005\u00c5 and an isotropic displacement parameter Uiso(H) = 1.2Uiso(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698902100579X/cq2042sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902100579X/cq2042Isup2.hklStructure factors: contains datablock(s) I. DOI: 2063269CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "ARs) make them an attractive target for selective and persistent modulation of neuronal excitability. However, the availability of selective modulators targeting \u03b4-GABAARs remains limited. AA29504 -phenyl]-carbamic acid ethyl ester), an analog of K+ channel opener\u00a0retigabine, acts as an agonist and a positive allosteric modulator (Ago-PAM) of \u03b4-GABAARs. Based on electrophysiological studies using recombinant receptors, AA29504 was found to be a more potent and effective agonist in \u03b4-GABAARs than in \u03b32-GABAARs. In comparison, AA29504 positively modulated the activity of recombinant \u03b4-GABAARs more effectively than \u03b32-GABAARs, with no significant differences in potency. The impact of AA29504's efficacy- and potency-associated GABAAR subtype selectivity on radioligand binding properties remain unexplored. Using [3H]4'-ethynyl-4-n-propylbicycloorthobenzoate ([3H]EBOB) binding assay, we found no difference in the modulatory potency of AA29504 on GABA- and THIP -induced responses between native forebrain GABAARs of wild type and \u03b4 knock-out mice. In recombinant receptors expressed in HEK293 cells, AA29504 showed higher efficacy on \u03b4- than \u03b32-GABAARs in the GABA-independent displacement of [3H]EBOB binding. Interestingly, AA29504 showed a concentration-dependent stimulation of [3H]muscimol binding to \u03b32-GABAARs, which was absent in \u03b4-GABAARs. This was explained by AA29504 shifting the low-affinity \u03b32-GABAAR towards a higher affinity desensitized state, thereby rising new sites capable of binding GABAAR agonists with low nanomolar affinity. Hence, the potential of AA29504 to act as a desensitization-modifying allosteric modulator of \u03b32-GABAARs deserves further investigation for its promising influence on shaping efficacy, duration and plasticity of GABAAR synaptic responses.The unique pharmacological properties of \u03b4-containing \u03b3-aminobutyric acid type A receptors (\u03b4-GABA AR), members of the Cys-loop ligand-gated ion channels superfamily, are the major sites for fast-acting synaptic inhibition in the mammalian brain pyridin-3-yl]benzamide), a selective positive allosteric modulator of \u03b14/6\u03b2\u03b4 receptors, is a widely used pharmacological tool to probe \u03b4-GABAesponses , 22. It lability . AA29504,2-apyridtrations \u201327. In rtrations , 25, 28.subtypes \u201327. AA29le cells , 25. In potency , 27. How potency . Earlierce versa . Hence, ARs. The selectivity of AA29504 to \u03b4-GABAARs was confirmed using wild-type (WT) and \u03b4 subunit knockout (\u03b4KO) C57BL/6\u00a0J mice forebrains, as well as recombinant receptors expressed in human embryonic kidney 293 (HEK293) cell line. Radioligands employed , a non-competitive blocker of GABA-gated chloride channel [3H]GABA, and [3H]muscimol, a universal GABAAR agonist with exceptionally high affinity to \u03b4-GABAARs Muscimol (22\u00a0Ci/mmol) and [3H]EBOB (48\u00a0Ci/mmol) were purchased from PerkinElmer Life and Analytical Sciences . [3H]GABA (30\u00a0Ci/mmol) was purchased from Moravek Biochemicals . Unlabeled GABA and picrotoxin were from Sigma Chemicals Co. . AA29504 and THIP were from Tocris Biosciences .muscimol (2\u00a0nM) and [3H]GABA (5\u00a0nM) were measured in assay buffer at room temperature (22 \u00baC) in a total volume of 300\u00a0\u00b5l. Individually pooled triplicate membrane samples were incubated with shaking for 15\u00a0min. The effect of AA29504 on binding was determined in the presence of various concentrations of AA29504. Non-specific binding was determined in the presence of 100\u00a0\u00b5M GABA. The incubation was terminated by filtration of the samples with a Brandel Cell Harvester onto Whatman GF/B filters .The binding of [3H]muscimol (2\u00a0nM) binding to the membrane homogenates were as follows: WT , \u03b4KO , \u03b11\u03b22\u03b32 , \u03b16\u03b22\u03b32 and \u03b16\u03b22\u03b4 . For [3H]GABA (5\u00a0nM) the binding values were: WT and \u03b4KO .The samples were rinsed twice with 4\u20135\u00a0ml of ice-cold assay buffer. Filtration and rinsing steps took a total time of 15\u00a0s. The filters were air-dried and immersed in 3\u00a0ml of Optiphase HiSafe 3 scintillation fluid and vortexed. The radioactivity was determined in a Wallac model 1410 liquid scintillation spectrometer . The average specific counts per minute (CPM) and % specific [3H]muscimol binding was measured essentially as described by Benkherouf et al. [3H]muscimol was performed essentially as described by Uusi-Oukari and Korpi [3H]muscimol (1\u201350\u00a0nM) at room temperature (22 \u00baC) for 60\u00a0min in the absence and presence of 10\u00a0\u00b5M AA29504. Non-specific binding was determined in the presence of 100\u00a0\u00b5M GABA. The incubation was terminated by filtration and the radioactivity of the air-dried filters was measured using a scintillation spectrometer as described above.The effect of AA29504 on the association and dissociation of [f et al. . Saturatnd Korpi . Triplic3H]EBOB binding was measured in [3H]EBOB assay buffer at room temperature (22 \u00baC) in a total volume of 400\u00a0\u00b5l in the absence and presence of various concentrations of GABA or THIP with and without 10\u00a0\u00b5M AA29504. Triplicate samples were incubated with shaking for 2\u00a0h. Non-specific binding was determined in the presence of 100\u00a0\u00b5M picrotoxin. The incubations were terminated as described above for [3H]muscimol binding. The average CPM and % specific [3H]EBOB (1\u00a0nM) binding to the membrane homogenates were as follows:WT , \u03b4KO , \u03b16\u03b23\u03b32 , \u03b16\u03b23\u03b4 , \u03b16\u03b23: .The displacement of 1\u00a0nM EBOB displacement from 0.20\u2009\u00b1\u20090.11\u00a0mM to 15.8\u2009\u00b1\u20096.5\u00a0\u00b5M in WT mice (p\u2009<\u20090.01) and from 0.22\u2009\u00b1\u20090.05\u00a0mM to 12.1\u2009\u00b1\u20091.5\u00a0\u00b5M in \u03b4KO mice . There were no significant differences between WT and \u03b4KO mouse binding values in the effects of GABA or THIP on forebrain GABAARs in the absence or presence of AA29504 .[3H]EBOB binding to recombinant GABAARs expressed in HEK293 cells as an indicator for allosteric agonist activity. The receptor subunit combinations \u03b16\u03b23\u03b32, \u03b16\u03b23\u03b4, and \u03b16\u03b23 were selected to further examine the influence of \u03b3 and \u03b4 subunits on [3H]EBOB binding displacement by AA29504. The results indicate that AA29504 was able to displace [3H]EBOB binding to all three receptor subtypes in a concentration-dependent manner .We assessed the potential of AA29504 to directly displace HEBOB binARs expressed in WT and \u03b4KO mice. The binding of [3H]muscimol and [3H]GABA was measured at room temperature (22\u00a0\u00b0C) with increasing concentrations of AA29504, where individually pooled forebrain membrane samples were incubated for 15\u00a0min. Figure\u00a03H]muscimol and [3H]GABA binding to both WT and \u03b4KO mice forebrains. Two-way ANOVA followed by Tukey's post hoc analysis indicated that AA29504 was more potent in stimulating [3H]muscimol (p\u2009<\u20090.01) and [3H]GABA (p\u2009<\u20090.001) binding in \u03b4KO than in WT mice.We further examined the modulatory effects of AA29504 on the high-affinity agonist binding to native GABAARs expressed in WT and \u03b4KO mice forebrains, we compared the role of \u03b3 and \u03b4 subunits on\u00a0AA29504 modulation of [3H]muscimol binding in recombinant \u03b11\u03b22\u03b32, \u03b16\u03b22\u03b32, and \u03b16\u03b22\u03b4 receptors expressed in HEK293 cells. As illustrated in Fig.\u00a03H]muscimol binding to \u03b11\u03b22\u03b32 and \u03b16\u03b22\u03b32 receptor subtypes in a concentration-dependent manner (p\u2009<\u20090.05). In contrast, it had no significant effect on the binding to \u03b16\u03b22\u03b4 receptor subtype (Two-way ANOVA followed by Tukey\u2019s post hoc test).Under the same conditions performed for native GABA3H]muscimol binding is due to\u00a0alterations in receptor-ligand binding kinetics as we assessed the influence of AA29504 on [3H]muscimol association and dissociation rates in WT and \u03b4KO forebrain membranes. The results indicate that [3H]muscimol association at 22\u00a0\u00b0C was faster in \u03b4KO compared to WT in the absence of AA29504, where the calculated association rate constants Kon were 5.9\u2009\u00b1\u20090.6\u2009\u00d7\u2009108\u00a0M\u22121\u2009\u00d7\u2009min\u22121 and 2.2\u2009\u00b1\u20090.3\u2009\u00d7\u2009108\u00a0M\u22121\u2009\u00d7\u2009min\u22121, respectively . Co-incubation with AA29504 did not significantly affect [3H]muscimol Kon in either \u03b4KO (4.8\u2009\u00b1\u20090.9\u2009\u00d7\u2009108\u00a0M\u22121\u2009\u00d7\u2009min\u22121) or WT mice (3.9\u2009\u00b1\u20091.1\u2009\u00d7\u2009108\u00a0M\u22121\u2009\u00d7\u2009min\u22121) , but it notably increased the amount of specific radioligand binding in both mouse lines (p\u2009<\u20090.001). The increased binding was maximally 240\u2009\u00b1\u200923% of control binding without AA29504 in \u03b4KO mice, significantly higher than that in WT mice (166\u2009\u00b1\u20095% of control) than in WT (0.38\u2009\u00b1\u20090.02\u00a0min\u22121) , reflecting a faster radioligand dissociation in the former mouse line . However, no evident effects were observed with AA29504 on [3H]muscimol.Similar to the association, the dissociation rate constant Koff in \u03b4KO (0.64\u2009\u00b1\u20090.05\u00a0min\u22121) and WT forebrains (0.43\u2009\u00b1\u20090.02\u00a0min\u22121) , in the absence (control) or presence of AA29504, all tested groups except control WT membranes were best fit to the one-site binding model. Binding to control WT membranes was best fit to the two-site binding model , displaying two binding affinities at distinguishable receptor densities (Table (1)\u2009+\u2009Bmax(2)) in WT membranes, however, was equivalent to control \u03b4KO Bmax (no AA29504). The presence of 10\u00a0\u00b5M AA29504 in WT membranes rendered [3H]muscimol binding more favorable to one-site model as it displayed a single apparent affinity that was intermediate between the affinities obtained in control WT membranes. On the other hand, AA29504 significantly decreased the equilibrium dissociation constant (KD) reflecting an enhancement of [3H]muscimol binding affinity in \u03b4KOs (p\u2009<\u20090.001). Moreover, AA29504 increased [3H]muscimol Bmax in \u03b4KO (p\u2009<\u20090.05) as well as WT mouse lines (p\u2009<\u20090.01) (Two-way ANOVA followed by Tukey's post hoc test). The calculated [3H]muscimol Bmax and KD values are summarized in Table As a probable mechanism for AA29504-induced stimulation of EBOB binding to recombinant GABAARs in this study, and similar to the earlier findings with electrophysiological measurements [3H]EBOB directly (in the absence of GABA) in the former subtype and dissociation (Koff) rate constants in either mouse line. Hence, the link between AA29504-induced stimulation of [3H]muscimol binding and alterations in receptor-ligand binding kinetics was not established.Modulating GABAed state , 65. Thu-GABAARs , 67 and -GABAARs . DespiteARs muscimol saturation analysis revealed AA29504-induced GABAAR shift to the high-affinity states. In WT mice, [3H]muscimol displayed two high-affinity receptor populations in the absence of AA29504: low-nanomolar (KD\u2009=\u20094.1\u2009\u00b1\u20093.6\u00a0nM) for \u03b4-GABAARs as earlier reported muscimol [AR population was suggested to rise from desensitized \u03b32-GABAARs\u00a0[3H]muscimol binding, especially in the\u00a0forebrain region muscimol binding EBOB radioligand as a\u00a0unique probe for assessing drug enhancement of GABAAR function, we demonstrated for the first time the non-differential AA29504 modulatory potency on native GABAARs expressed in WT and \u03b4KO C57BL/6J mice. We further displayed AA29504\u2019s GABA-independent activity on recombinant GABAARs expressed in HEK293 cells, indicating higher selective agonist efficacy on \u03b4-GABAARs in relation to \u03b32-GABAARs. Interestingly, AA29504 showed a concentration-dependent stimulation of GABAA agonist binding to \u03b32 GABAARs but\u00a0not to \u03b4-GABAARs. This newly revealed selective modulation by AA29504 is attributed to its ability to shift the low-affinity \u03b32-GABAARs towards a higher affinity desensitized state, thereby rising new sites capable of binding GABAAR agonists with low nanomolar affinity. Hence, the potential of AA29504 to act as a desensitization-modifying allosteric modulator (DAM) of \u03b32-GABAARs deserves further investigation for its promising influence on shaping efficacy, duration and plasticity of GABAAR synaptic responses [This study sheds light on AA29504's modulatory activity, its direct actions and interactions with agonists in GABAesponses , 84, 85."} +{"text": "The mol\u00adecular and crystal structures were studied and a Hirshfeld surface analysis undertaken for the title benzo\u00adthia\u00adzine derivative, which has potential non-steroidal anti-inflammatory activity. 22H22ClN3O4S, which has potential non-steroidal anti-inflammatory activity, the benzo\u00adthia\u00adzine and cyclo\u00adhexenone rings both adopt a distorted sofa conformation while the 4H-pyrane ring adopts a very flattened sofa conformation. The two bicyclic fragments are skewed to each other, with the dihedral angle between their least-squares planes being 72.8\u2005(1)\u00b0. In the crystal, the mol\u00adecules form a hydrogen-bonded chain parallel to the a axis due to N\u2014H\u22efO and N\u2014H\u22efCl hydrogen bonds. Neighbouring chains are linked by C\u2014H\u22efN, C\u2014H\u22efO and \u03c0\u2013\u03c0 stacking inter\u00adactions. Hirshfeld surface analysis was used to investigate the importance of the different types of inter\u00admolecular inter\u00adactions whose contributions are: H\u22efH = 44.7%, O\u22efH/H\u22efO = 21.8%, N\u22efH/H\u22efN = 11.9%, C\u22efH/H\u22efC = 9.5%, Cl\u22efH/H\u22efCl = 7.2%. Parts of the mol\u00adecule, viz. the phenyl ring and the ethyl side chain, are equally disordered over two sets of sites.In the title compound, C H-benzo[c]thia\u00adzine 2,2-dioxide moiety and its derivatives have been the focus of chemists and pharmacologists for decades of the \u2018oxicame\u2019 group , \u0398 = 52.5\u2005(1)\u00b0, \u03a8 = 20.3\u2005(1)\u00b0. The S1 and C8 atoms deviate from the least-squares plane of the remaining atoms of the ring by 0.863\u2005(6) and 0.244\u2005(2)\u2005\u00c5 respectively. The phenyl ring of the benzo\u00adthia\u00adzine fragment is disordered over two positions (A and B) with equal occupancy. The partially saturated carbocycle has the same conformation as the hydro\u00adthia\u00adzine ring, with puckering parameters of S = 0.67\u2005(1), \u0398 = 41.9\u2005(1)\u00b0, \u03a8 = 11.8\u2005(1)\u00b0. The deviations of the C13 and C14 atoms from the least-squares plane of the remaining atoms in the ring are 0.717\u2005(2) and 0.132\u2005(2)\u2005\u00c5, respectively. The 4H-pyran ring adopts a very flattened sofa conformation with puckering parameters of S = 0.11\u2005(1), \u0398 = 59.3\u2005(1)\u00b0, \u03a8 = 3.2\u2005(1)\u00b0, where the C9 atom deviates by 0.118\u2005(2)\u2005\u00c5 from the plane of the remaining atoms in this ring. The C8\u2014C9 bond is elongated to 1.525\u2005(3)\u2005\u00c5 [the mean value \u00b0]. The presence of the vicinal substituents on the 4H-pyran moiety results in an elongation of the C16\u2014C17 bond to 1.347\u2005(3)\u2005\u00c5 .The di\u00adhydro\u00adthia\u00adzine ring of compound on Fig.\u00a02 with puc4 form hydrogen-bonded chains parallel to the a axis 1\u00a0+\u00a0x, y, z; Table\u00a01In the crystal, mol\u00adecules of is Fig.\u00a03 due to NH-pyran rings of mol\u00adecules belonging to neighbouring layers are found [the distance between ring planes is 3.38\u2005(1)\u2005\u00c5 and the plane shift is 1.247\u2005(1)\u2005\u00c5]. Mol\u00adecules are arranged in a head-to-tail manner in both types of stacking dimers. Additional C\u2014H\u22efN and C\u2014H\u22efO hydrogen-bonding inter\u00adactions of a weak nature is provided by O\u22efH/H\u22efO inter\u00adactions , while the highest contribution is from H\u22efH contacts (44.7%). The contributions of N\u22efH/H\u22efN (11.9%), C\u22efH/H\u22efC (9.5%) and Cl\u22efH/H\u22efCl (7.2%) inter\u00adactions are similar, but the presence of sharp spikes on the fingerprint plot containing only N\u22efH/H\u22efN or Cl\u22efH/H\u22efCl inter\u00adactions suggests that the latter contacts are much stronger.All of the hydrogen bonds and short contacts of the title compound are evident on the two-dimensional fingerprint plot presented in Fig.\u00a05ns Fig.\u00a05c, whileet al., 2016et al., 2014aet al., 2016et al., 2018et al., 2014bet al., 2017et al., 2018A search of the Cambridge Structural Database , malono\u00adnitrile (2) and 5,5-di\u00admethyl\u00adcyclo\u00adhexane-1,3-dione (3) was dissolved in 20\u2005ml of i-PrOH and then tri\u00adethyl\u00adamine (0.1\u2005mol%) was added ; colourless crystals; m.p. > 523\u2005K.A mixture of 4-chloro-1-ethyl-1ed Fig.\u00a01. The mixsp3\u2014Csp3 = 1.54\u2005\u00c5 for the ethyl side chain C21\u2014C22; Csp2\u2014Csp2 = 1.38\u2005\u00c5 for the phenyl ring C1\u2013C6), and with an equal occupancy for the two sets of sites. All hydrogen atoms were located in difference-Fourier maps. They were included in calculated positions and treated as riding with C\u2014H = 0.96\u2005\u00c5, Uiso(H) = 1.5Ueq(C) for methyl groups and with Car\u2014H = 0.93\u2005\u00c5, Csp3\u2014H = 0.97\u2005\u00c5, Uiso(H) = 1.2Ueq(C) for all other hydrogen atoms. The hydrogen atoms of the amino group were refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021002085/wm5596sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021002085/wm5596Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021002085/wm5596Isup3.cmlSupporting information file. DOI: 2064493CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Transforming growth factor beta (TGF\u2010\u03b2) plays an important role in the viral liver disease progression via controlling viral propagation and mediating inflammation\u2010associated responses. However, the antiviral activities and mechanisms of TGF\u2010\u03b2 isoforms, including TGF\u2010\u03b21, TGF\u2010\u03b22 and TGF\u2010\u03b23, remain unclear. Here, we demonstrated that all of the three TGF\u2010\u03b2 isoforms were increased in Huh7.5 cells infected by hepatitis C virus (HCV), but in turn, the elevated TGF\u2010\u03b2 isoforms could inhibit HCV propagation with different potency in infectious HCV cell culture system. TGF\u2010\u03b2 isoforms suppressed HCV propagation through interrupting several different stages in the whole HCV life cycle, including virus entry and intracellular replication, in TGF\u2010\u03b2/SMAD signalling pathway\u2013dependent and TGF\u2010\u03b2/SMAD signalling pathway\u2013independent manners. TGF\u2010\u03b2 isoforms showed additional anti\u2010HCV activities when combined with each other. However, the elevated TGF\u2010\u03b21 and TGF\u2010\u03b22, not TGF\u2010\u03b23, could also induce liver fibrosis with a high expression of type I collagen alpha\u20101 and \u03b1\u2010smooth muscle actin in LX\u20102 cells. Our results showed a new insight into TGF\u2010\u03b2 isoforms in the HCV\u2010related liver disease progression. The expressions of TGF\u2010\u03b2 isoforms are promoted by HCV infection in cultured hepatocytes and in livers of HCV\u2010infected patients, which might be caused by HCV\u2010induced endoplasmic reticulum stress, unfolded protein response activation or HCV core expression.22.1The human hepatoma cell line Huh7.5 cells and the plasmid pFL\u2010J6/JFH/JC1 (HCV\u20102a) containing the full\u2010length chimeric HCV complementary DNA (cDNA) were kindly provided by Vertex Pharmaceuticals Inc.2.2TGF\u2010\u03b21 (100\u201021\u201010\u00a0\u03bcg), TGF\u2010\u03b22 (100\u201035B\u201010\u00a0\u03bcg) and TGF\u2010\u03b23 (100\u201036E\u201010\u00a0\u03bcg) proteins were purchased from PeProtech Inc (Peprotech). All TGF\u2010\u03b2 isoforms were dissolved according to the manufacturer's protocol and the cell viability after TGF\u2010\u03b2 isoforms treatment was determined by a methyl thiazolyl tetrazolium assay.2.3The plasmids pcDNA3.1(+)\u2010TGF\u2010\u03b21 expressing TGF\u2010\u03b21, pcDNA3.1(+)\u2010TGF\u2010\u03b23 expressing TGF\u2010\u03b23, pcDNA3.1(+)\u2010TGF\u2010\u03b21\u2010His expressing TGF\u2010\u03b21 with His tag at the C\u2010terminal, pcDNA3.1(+)\u2010TGF\u2010\u03b23\u2010His expressing TGF\u2010\u03b23 with His tag at the C\u2010terminal and the part truncated mutant plasmids of pcDNA3.1(+)\u2010TGF\u2010\u03b21\u2010His or pcDNA3.1(+)\u2010TGF\u2010\u03b23\u2010His were generated by amplification of cDNA from Huh7.5 cell using primers shown in Table\u00a02.4\u2212\u0394\u0394CT method. Glyceraldehyde 3\u2010phosphate dehydrogenase (GAPDH) was served as the internal control for the quantification. The primer and probe sequences used for quantitative real\u2010time reverse transcriptase polymerase chain reaction (qRT\u2010PCR) are provided in Table\u00a0The cellular RNAs harvested by RNeasy Mini Kit (QIAGEN) from Huh7.5 cells were detected using the AgPath\u2010ID One\u2010Step RT\u2010PCR Kit (Applied Biosystems). Fluorescent signals were identified with 7500 fast real\u2010time PCR system (Applied Biosystems) according to the procedure recommended by the manufacturer. HCV RNA level was calculated according with the 2TGFB1, TGFB2, TGFB3, type I collagen alpha 1 (COL1A1) or alpha\u2010smooth muscle actin (\u03b1\u2010SMA) were quantified using GoTaq qPCR Master Mix according to the procedure. The primers are shown in Table\u00a0For mRNA quantitation by SYBR Green method, total RNAs were reverse\u2010transcribed with PrimeScript RT Master Mix , and the mRNAs of 2.5Huh7.5 cells were treated with HCV viral stock at different multiplicity of infection for 72\u00a0hours or at MOI\u00a0=\u00a00.2 for 0, 3\u00a0days or 6\u00a0days, and then, intracellular mRNAs or proteins were extracted and quantified by qRT\u2010PCR or Western blot (WB) analysis.2.6Huh7.5 cells were inoculated with HCV viral stock (MOI\u00a0=\u00a00.5) and simultaneously treated with TGF\u2010\u03b2 isoforms or sofosbuvir. At 72\u00a0hours, intracellular proteins were harvested by the CytoBuster Protein Extraction Reagent containing 1\u00a0mmol/L protease inhibitor cocktail (Roche Applied Science) and HCV core protein level in cells was determined by WB analysis.\u22121 to 10\u22128 and simultaneously treated with 0.5\u00a0nmol/L of TGF\u2010\u03b21, TGF\u2010\u03b22 and TGF\u2010\u03b23. At 72\u00a0hours post\u2010infection, HCV\u2010positive wells were examined using a microscope after immunostaining against HCV core protein.For measuring HCV infectivity, Huh7.5 cells were incubated with HCV viral stock at 10\u2010fold dilutions ranged from 10de novo HCV particle production, Huh7.5 cells were inoculated with HCV viral stock (MOI\u00a0=\u00a00.7) and 0.5\u00a0nmol/L of TGF\u2010\u03b21, TGF\u2010\u03b22 or TGF\u2010\u03b23 for 24\u00a0hours. The newly released infectious virus particles were collected at 48\u00a0hours and then quantified by 50% tissue culture infective dose (TCID50) assay. The infectivity titre was shown as TCID50 per mL.To identify the effects of TGF\u2010\u03b2 isoforms on GS4.3 cells were treated with 0.5\u00a0nmol/L of TGF\u2010\u03b2 isoforms or sofosbuvir (0.5\u00a0\u03bcmol/L) for 72\u00a0hours. Then, intracellular proteins were extracted by PER and HCV NS3 protein level was detected by WB analysis.2.7To evaluate the antiviral effective stages of TGF\u2010\u03b2 isoforms, Huh7.5 cells were pre\u2010treated with TGF\u2010\u03b2 isoforms (0.5\u00a0nmol/L) for 2\u00a0hours followed by HCV infection for 2\u00a0hours, simultaneously treated with TGF\u2010\u03b2 isoforms (0.5\u00a0nmol/L) and HCV for 2\u00a0hours, or infected with HCV (MOI\u00a0=\u00a00.5) for 2\u00a0hours followed by TGF\u2010\u03b2 isoforms (0.5\u00a0nmol/L) 2\u00a0hours treatment. At 72\u00a0hours, intracellular HCV core protein level was detected by WB analysis. In order to further characterize the inhibition stages, a time\u2010of\u2010addition assay was performed. In brief, Huh7.5 cells were inoculated with HCV (MOI\u00a0=\u00a00.5) at 37\u00b0C for 2, 4, 6, 8\u00a0hours or 10\u00a0hours, and then, the HCV\u2010containing medium was replaced by TGF\u2010\u03b2 isoforms (0.5\u00a0nmol/L) or sofosbuvir (0.5\u00a0\u03bcmol/L) containing medium. After 2\u00a0hours of treatment, the medium was replaced by fresh medium. At 72\u00a0hours, HCV core protein level in cells was determined by WB analysis.2.8HCV viral stock (MOI\u00a0=\u00a00.5) was pre\u2010treated with 0.2\u00a0nmol/L TGF\u2010\u03b21, 0.5\u00a0nmol/L TGF\u2010\u03b22 or 0.5\u00a0nmol/L TGF\u2010\u03b23 at 37\u00b0C and used to infect cells after 25\u2010fold dilution of the inoculum. Infection with untreated HCV stock was performed in parallel in the presence of TGF\u2010\u03b2 isoforms at indicated concentrations. At 72\u00a0hours, intracellular proteins were harvested and HCV core protein level was determined by WB.2.9Huh7.5 cells were incubated with HCV viral stock (MOI\u00a0=\u00a00.5) for 16\u00a0hours, and then, the cells were treated with TGF\u2010\u03b2 isoforms (0.5\u00a0nmol/L), sofosbuvir (0.5\u00a0\u03bcmol/L) or 0.1% BSA for 48\u00a0hours. The culture medium was replaced by adding fresh culture medium. After 24\u00a0hours, the cell viability was tested using MTT assay, the culture medium was collected for incubation to na\u00efve Huh7.5 cells, and intracellular RNA was extracted and quantified by qRT\u2010PCR. Meanwhile, na\u00efve Huh7.5 cells were incubated with above culture supernatants for 72\u00a0hours. Then, the intracellular RNA of Huh7.5 cells was extracted and quantified by qRT\u2010PCR.2.10q value was calculated with the improved B\u00fcrgi formula (Jin's equation), with q\u00a0<\u00a00.85, 0.85\u00a0~\u00a01.15, and >1.15 indicating antagonism, addition and synergy, respectively.Huh7.5 cells were inoculated with HCV viral stock (MOI\u00a0=\u00a00.5) and simultaneously treated with 0.04\u00a0nmol/L TGF\u2010\u03b21, 0.05\u00a0nmol/L TGF\u2010\u03b22, 0.1\u00a0nmol/L TGF\u2010\u03b23 alone, in combination or double concentration (2\u00d7), or sofosbuvir (0.5\u00a0\u03bcmol/L). At 72\u00a0hours, intracellular proteins were extracted and determined by WB analysis. To evaluate their combined effect, the combination index (CI) showed with 2.11Huh7.5 cells were incubated with T\u03b2RI or T\u03b2RII kinase inhibitors (20\u00a0\u03bcmol/L LY2109761 or 50\u00a0\u03bcmol/L RepSox) for 6\u00a0hours, and then, TGF\u2010\u03b2 isoforms (0.5\u00a0nmol/L) and HCV (MOI\u00a0=\u00a00.5) were simultaneously added. After 72\u00a0hours incubation, intracellular HCV core protein was determined by WB analysis.2.12Huh7.5 cells were transfected with TGF\u2010\u03b21, TGF\u2010\u03b21\u2010His, TGF\u2010\u03b23, TGF\u2010\u03b23\u2010His, the mutants of TGF\u2010\u03b2 plasmids or plasmid control pcDNA3.1(+) applying HD transfection reagent (Promega), or transfected with siRNA for T\u03b2RI or SMAD2/3 or negative control siRNA\u2010A applying RNAiMAX transfection reagent (Invitrogen). After 48\u00a0hours, the cells were sub\u2010cultured and part of intracellular proteins was harvested to detect His tag or corresponding protein expression with WB. After 24\u00a0hours, cells were infected with HCV (MOI\u00a0=\u00a00.5) or added TGF\u2010\u03b2 isoforms (0.5\u00a0nmol/L) and HCV (MOI\u00a0=\u00a00.5). After 48\u00a0hours, intracellular proteins were extracted to detect HCV core protein level by WB.2.13COL1A1 and \u03b1\u2010SMA mRNA was quantified with qRT\u2010PCR.LX\u20102 cells were cultured in high glucose DMEM without FBS for 12\u00a0hours. Then, the cell supernatant was replaced by fresh medium with 2% FBS containing 0.5\u00a0nmol/L TGF\u2010\u03b2 isoforms. After 24\u00a0hours, intracellular RNA was extracted and 2.14The Western blot analysis was performed as previously described.2.15t test; P\u00a0<\u00a00.05 was considered as statistically significant.The data were presented as means\u00a0\u00b1\u00a0standard deviation (SD) more than three independent experiments. Statistical analyses were performed in GraphPad Prism 7 (GraphPad) using ANOVA analysis followed by the Student\u2019s 33.1In infectious HCV cell culture (HCVcc) system, the three TGF\u2010\u03b2 isoforms at mRNA Figure\u00a0, left anDe novo HCV production, Huh7.5 cells were infected with HCV and simultaneously treated with TGF\u2010\u03b2 isoforms for 24\u00a0hours, and newly released infectious virus particles were collected at 48\u00a0hours and then quantified by TCID50 assay. De novo produced HCV yielded a viral titre of about 104.17 TCID50 per mL, whereas treatment with TGF\u2010\u03b2 isoforms significantly decreased the TCID50 of the De novo produced HCV \u2010TGF\u2010\u03b22 could not well express TGF\u2010\u03b22 in Huh7.5 cells as TGF\u2010\u03b21 and TGF\u2010\u03b23 did. Therefore, we only test the ant\u2010HCV effects caused by overexpressing TGF\u2010\u03b21 and TGF\u2010\u03b23 to investigate which regions or amino acid residues of TGF\u2010\u03b2 isoforms were responsible for their anti\u2010HCV capacities. Overexpressing TGF\u2010\u03b21 and TGF\u2010\u03b23 with His tag at C\u2010terminal had equal anti\u2010HCV effects as TGF\u2010\u03b21 and TGF\u2010\u03b23, respectively Figure\u00a0, suggestCys77 or Cys109 in TGF\u2010\u03b2 isoforms is important to form homodimer for activation of their signalling activities.The amino acid residues Arg25, Val92 and Arg94 of TGF\u2010\u03b21 and TGF\u2010\u03b23 are responsible for their high\u2010affinity interaction with T\u03b2RII, once they are exchanged with Lys25, Ile92 and Lys94, respectively, TGF\u2010\u03b2 signalling activities of TGF\u2010\u03b21 and TGF\u2010\u03b23 are reduced.The \u03b1\u2010helix 3 of TGF\u2010\u03b2 isoforms is response for their binding surface for T\u03b2RI and truncating \u03b1\u2010helix 3 of TGF\u2010\u03b22 suppresses its signalling activity.Interestingly, by blocking the TGF\u2010\u03b2 receptor kinase activity or silencing SMAD2/3, we presumed the anti\u2010HCV activities of TGF\u2010\u03b21 and TGF\u2010\u03b22 might be through TGF\u2010\u03b2/SMAD signalling pathway, while TGF\u2010\u03b23 inhibited HCV replication independent on TGF\u2010\u03b2/SMAD signalling pathway Figure\u00a0. This di3.5COL1A1 and \u03b1\u2010SMA mRNA expression ; Data curation (lead); Methodology (lead); Writing\u2010original draft ; Writing\u2010review & editing (lead). Jian\u2010Rui Li: Data curation (lead); Methodology (lead). Hu Li: Methodology (supporting); Writing\u2010review & editing (supporting). Jia\u2010Li Tan: Data curation (supporting); Methodology (supporting); Writing\u2010review & editing (supporting). Mei\u2010Xi Wang: Writing\u2010review & editing (supporting). Nan\u2010Nan Liu: Writing\u2010review & editing (supporting). Rong\u2010Mei Gao: Data curation (supporting); Methodology (supporting). Hai\u2010Yan Yan: Data curation (supporting); Methodology (supporting). Xue\u2010Kai Wang: Writing\u2010review & editing (supporting). Biao Dong: Methodology (supporting). Yu\u2010Huan Li: Methodology (supporting). Zonggen Peng: Conceptualization (lead); Data curation (lead); Formal analysis (lead); Funding acquisition (lead); Methodology (lead); Project administration (lead); Writing\u2010review & editing (lead)."} +{"text": "Two crystallographically independent mol\u00adecules are present in the asymmetric unit. O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds form rings and chains and \u03c0\u2013\u03c0 stacks further connect mol\u00adecules in the crystal. 14H11NO2, with virtually identical geometries. The carbazole units are planar. The hy\u00addroxy group at position 1, carbaldehyde group at position 2, and methyl group at position 8 (with the exception of two H atoms) are coplanar with the attached benzene rings. The dihedral angle between the two benzene rings is 2.20\u2005(9)\u00b0 in mol\u00adecule A and 2.01\u2005(9)\u00b0 in mol\u00adecule B. The pyrrole ring makes dihedral angles of 0.82\u2005(10) and 1.40\u2005(10)\u00b0 [0.84\u2005(10) and 1.18\u2005(10)\u00b0 in mol\u00adecule B] with the (\u2013CH3)-substituted and (\u2013OH and \u2013CHO) substituted benzene rings, respectively. The mol\u00adecular structure is stabilized by the intra\u00admolecular O\u2014H\u22efO hydrogen bonds, while the crystal structure features N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. A range of \u03c0\u2013\u03c0 contacts further stabilizes the crystal structure.Two crystallographically independent mol\u00adecules are present in the asymmetric unit of the title compound, C Murraya koenigii spreng (Rutaceae) is a rich source of carbazole alkaloids \u00b0 in mol\u00adecule B. The pyrrole ring makes dihedral angles of 0.82\u2005(10) and 1.40\u2005(10)\u00b0 for mol\u00adecule A and 0.84\u2005(10) and 1.18\u2005(10)\u00b0 for mol\u00adecule B with the methyl-substituted and hydroxide/carbaldehyde-substituted benzene rings, respectively. The compound exhibits intra\u00admolecular O\u2014H\u22efO hydrogen bonding between the hydroxide and aldehyde groups ring motifs, have previously been observed and N2\u2014H2\u22efO2 hydrogen bonds. A C14\u2014H14\u22efO3 hydrogen bond is also present. A range of \u03c0\u2013\u03c0 contacts is also observed = 3.4604\u2005(13)\u2005\u00c5, Cg1\u22efCg3 = 3.4896\u2005(13)\u2005\u00c5 and Cg7\u22efCg9 = 3.6279\u2005(13)\u2005\u00c5, where Cg1, Cg2, Cg3, Cg7 and Cg9 are the centroids of the N1/C7/C6/C10/C9, C2\u2013C7, C8\u2013C13, N2/C21/C20/C24/C23 and C22\u2013C27 rings, respectively.In the crystal, mol\u00adecules are connected into chains parallel to the s Table\u00a01. Both cred Fig.\u00a04. The diset al., 2016H-carbazole-2-carbaldehyde gave two hits, viz. 2,2,10-trimethyl-2,3-di\u00adhydro\u00adpyranocarbazol-4(11H)-one ethanone in dry benzene (25\u2005ml) was added slowly and the reaction mixture was allowed to stir for another 36\u2005h. The reaction was monitored by TLC. After completion of the reaction, benzene was removed in vacuo and the contents in the flask were transferred to a beaker containing water. It was neutralized with dilute HCl, filtered, washed with water and dried to get crude 1-hy\u00addroxy-8-methyl-9H-carbazole-2-carbaldehyde. It was purified by column chromatography over silica using petroleum ether:ethyl acetate (95:5) as eluant. The brown pure product obtained was recrystallized using glacial acetic acid , m.p. 414\u2005K was washed with dry benzene and taken into a round-bottom flask containing dry benzene (100\u2005ml). The flask was kept in an ice bath under stirring. Ethyl formate (8\u2005ml) was added dropwise to the solution over a period of 10 minutes. Then 8-methyl-2,3,4,9-tetra\u00adhydro-1\u2005K Fig.\u00a05.D and H3A were located in a difference-Fourier map and freely refined. The remaining hydrogen atoms were placed in calculated positions with C\u2014H bond distances of 0.93\u2005\u00c5 (aromatic H), or 0.96\u2005\u00c5 (methyl H) and were refined with anisotropic displacement parameters 1.2 and 1.5 times that of the parent carbon atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021007210/yy2001sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021007210/yy2001Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021007210/yy2001Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021007210/yy2001Isup4.cmlSupporting information file. DOI: 1540679CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "We report here the following nucleotide incorporations opposite Ua by various DNA polymerases: DNA polymerase \u03b1, dATP and dGTP (dATP > dGTP); DNA polymerase \u03b4, dATP; DNA polymerase \u03b6, dATP; Kf exo\u2212, dATP; Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4), dGTP and dATP (dGTP > dATP); and DNA polymerase \u03b7, dCTP, dGTP, dATP, and dTTP (dCTP > dGTP > dATP > dTTP). DNA polymerases \u03b2 and \u03b5 were blocked by Ua. Elongation by DNA polymerases \u03b4 and \u03b6 stopped after inserting dATP opposite Ua. Importantly, the elongation efficiency to full-length beyond Ua using DNA polymerase \u03b7 and Dpo4 were almost the same as that of natural DNA.Urea (Ua) is produced in DNA as the result of oxidative damage to thymine and guanine. It was previously reported that Klenow fragment (Kf) exoThe online version contains supplementary material available at 10.1186/s41021-022-00236-3. Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4) have not been analyzed to date. Herein, we describe our studies of nucleotide incorporations and translesion synthesis in relation to Ua with DNA polymerases \u03b1, \u03b4, \u03b5, \u03b7, \u03b6 and Dpo4. Also, we confirmed nucleotide incorporation and extension in relation to Ua using Kf exo\u2212 and DNA polymerase \u03b2.DNA damage is a major cause of cell death, mutations, cancer, neurological disease, and aging. Urea (Ua) is produced in DNA by ionizing radiation due to o\u2212 was purchased from Fermentas . Yeast DNA polymerase \u03b6 was purchased from Enzymax . Human DNA polymerase \u03b4 [Saccharomyces cerevisiae DNA polymerase \u03b5 [T4 polynucleotide kinase was purchased from New England Biolabs . T4 DNA ligase was purchased from Takara . Calf thymus DNA polymerase \u03b1 and human DNA polymerase \u03b2 were purchased from Chimerx . Klenow fragment exonucleasemerase \u03b4 , human Dmerase \u03b4 , and Sacmerase \u03b5 were purThe DNA template , Alexa 680-labeled 15-mer primer (5\u2032-Alexa680-TATTGATTGTGAATT-3\u2032), 6-mer oligonucleotide containing 8oxoG (5\u2032-CTT8oxoGAA-3\u2032), 13-mer oligonucleotide (5\u2032-TTCACAATCAATA-3\u2032), and 11-mer oligonucleotide (5\u2032-CTCATCAACAT-3\u2032) were constructed by Japan Bio Services Co., Ltd. .2 and KI [The 30-mer DNA template containing Ua (30-merUa) was constructed as follows. A 6-mer oligonucleotide containing 8oxoG (5\u2032-CTT8oxoGAA-3\u2032) was oxidized to a 6-mer oligonucleotide containing oxaluric acid (Oxa) using I2 and KI , then Ox2 and KI . This 6-2, 10\u2009mM NaCl, 45\u2009mM KCl, 1\u2009mM DTT, 100\u2009\u03bcg/mL bovine serum albumin (BSA); (for DNA polymerase \u03b2) 50\u2009mM Tris-HCl (pH\u20098.8), 10\u2009mM MgCl2, 1\u2009mM DTT, 100\u2009\u03bcg/mL BSA; (for DNA polymerase \u03b4) 50\u2009mM Tris-HCl (pH\u20097.4), 2\u2009mM MgCl2, 2\u2009mM DTT, 100\u2009\u03bcg/mL BSA; (for DNA polymerase \u03b5) 50\u2009mM Tris-HCl (pH\u20097.4), 8\u2009mM MgCl2, 2\u2009mM DTT, 100\u2009\u03bcg/mL BSA; (for DNA polymerases \u03b6 and \u03b7) 50\u2009mM Tris-HCl (pH\u20098.0), 2\u2009mM MgCl2, 5\u2009mM DTT, 100\u2009\u03bcg/mL BSA; (for Kf exo\u2212) 50\u2009mM Tris-HCl (pH\u20098.0), 5\u2009mM MgCl2, 1\u2009mM DTT, 100\u2009\u03bcg/mL BSA; (for Dpo4) 20\u2009mM HEPES (pH\u20096.5), 10\u2009mM MgCl2, 1\u2009mM MnCl2 100\u2009mM NaCl, 100\u2009\u03bcg/mL BSA.Polymerization reactions (5\u2009\u03bcL) were carried out using the following mixtures: (for DNA polymerase \u03b1) 40\u2009mM Tris-HCl (pH\u20098.0), 5\u2009mM MgCl\u2212 and Dpo4 contained 100 fmol of the template and 50 fmol of 5\u2032-Alexa680-labeled 15-mer primer. Other conditions and the concentrations of dNTPs and DNA polymerases are specified in the figure legends. Reactions were performed at 30\u2009\u00b0C for 30\u2009min for DNA polymerases \u03b1, \u03b2, \u03b4, \u03b5, \u03b6, Kf exo\u2212 and Dpo4, and at 37\u2009\u00b0C for 30\u2009min for DNA polymerase \u03b7. All reactions were stopped by adding 5\u2009\u03bcL of stop buffer , and 100\u2009\u03bcM rhodamine 6G). Aliquots (2.5\u2009\u03bcL) were subjected to electrophoresis in a denaturing 16% polyacrylamide (v/v) gel containing 8\u2009M urea at 30\u2009W for 90\u2009min. The fluorescence intensity of each n-mer band (In) was quantified using an Odyssey infrared imaging system from LI-COR for 5\u2032-Alexa 680-labeled products. The nucleotide incorporation efficiency was calculated using the formula: \u03a3 In (n\u2009\u2265\u200916) / \u03a3 In (n\u2009\u2265\u200915). The DNA synthesis efficiency was calculated using the formula: I30 / \u03a3 In (n\u2009\u2265\u200915).The reaction mixtures for DNA polymerases \u03b1, \u03b2, \u03b4, \u03b5, \u03b7, \u03b6, Kf exoDNA polymerase \u03b1 was used for in vitro nucleotide insertion and the analysis of primer extension of template oligonucleotides containing Ua. Under the reaction condition in which DNA polymerase \u03b1 inserted only dCTP opposite guanine Fig.\u00a0A, lane 8We investigated whether DNA polymerase \u03b1 extends the primer beyond Ua. DNA polymerase \u03b1 slightly elongated the 15-mer primer to 30-mer (3%) across the Ua lesion opposite Ua under the condition that incorporated only dCTP into guanine in the template Fig.\u00a0B, lane 7DNA polymerase \u03b6 predominantly inserted mainly dATP 3%) opposite Ua Fig.\u00a0A, lane 7% opposit\u2212 [\u2212, we re-analyzed nucleotide incorporation and translesion synthesis opposite Ua. The results of elongation efficiency by DNA polymerase \u03b2 beyond Ua -oxazolone (Oz) using DNA polymerase \u03b6 was approximately the same as that of natural DNA [DNA polymerase \u03b6 plays a critical role in an error-prone lesion bypass pathway \u201337. We pural DNA . AnalysiDNA polymerase \u03b7 can efficiently and accurately extend the primer beyond the cyclobutane pyrimidine dimer , as wellDpo4 is a thermostable translesion synthesis polymerase and elongated the primer to full-length across Ua were obtained . However, these two products equilibrate with each other and thus could not be isolated separately. In a previous report [289H370N103O176P29) was confirmed by ESI-MS (m/z 9000.727) and then was used as 30-merUa in our experiment.s report , Dubey, Additional file 2: Fig. S2. DNA synthesis across urea (Ua) by Kf exo\u2212. DNA synthesis in Fig. S2 was conducted under the same condition as in Fig. \u2212 (75 \u03bcU) was incubated with templates containing G (lane 3) or Ua (lane 4) and 100\u2009\u03bcM of each of the four dNTPs (lanes 1\u20134). Lanes 1 and 2 contained no enzyme and are negative controls. The background darkness of panel A is adjusted in Panel B. Fig. S3. DNA synthesis across urea (Ua) by DNA polymerase \u03b7. DNA synthesis in Fig. S3 was conducted under the same condition as in Fig. Ua lane and 100"} +{"text": "Substitution of CO by phosphanes influences the bond parameters more than replacing the C5H5 ligand by C5H4Cl.The syntheses and structures of the cymantrenes [(C 3] (Cp\u00a0= \u03c0-C5H5 or \u03c0-C5H4Cl) in the presence of the phosphanes PPh3 or PCy3 (Cy\u00a0= cyclo\u00adhex\u00adyl) and Ph2PCH2CH2PPh2 yields the substitution products [CpMn(CO)2PR3] (R\u00a0= Ph or Cy) and [CpMn(CO)(Ph2PCH2CH2PPh2)], namely, dicarbon\u00adyl(\u03b75-cyclo\u00adpenta\u00addien\u00adyl)(tri\u00adphenyl\u00adphosphane-\u03baP)manganese(I), [Mn(C5H5)(C18H15P)(CO)2], 1a, dicarbon\u00adyl(\u03b75-1-chloro\u00adcyclo\u00adpenta\u00addien\u00adyl)(tri\u00adphenyl\u00adphosphane-\u03baP)man\u00adganese(I), [Mn(C5H4Cl)(C18H15P)(CO)2], 1b, dicarbon\u00adyl(\u03b75-cyclo\u00adpenta\u00addien\u00adyl)(tri\u00adcyclo\u00adhexyl\u00adphosphane-\u03baP)manganese(I), [Mn(C5H5)(C18H33P)(CO)2], 2a, di\u00adcarbon\u00adyl(\u03b75-1-chloro\u00adcyclo\u00adpenta\u00addien\u00adyl)(tri\u00adcyclo\u00adhexyl\u00adphosphane-\u03baP)manganese(I), [Mn(C5H4Cl)(C18H33P)(CO)2], 2b, carbon\u00adyl(\u03b75-cyclo\u00adpenta\u00addien\u00adyl)manganese(I), [Mn(C5H5)(C26H24P2)(CO)], 3a, and carbon\u00adyl(\u03b75-1-chloro\u00adcyclo\u00adpenta\u00addien\u00adyl)manganese(I), [Mn(C5H4Cl)(C26H24P2)(CO)], 3b, The crystal structure determinations show a very small influence of the chlorine substitution and a moderate influence of the phosphane substitution on the bond lengths. The PR3 groups avoid being eclipsed with the C\u2014Cl bonds. All the com\u00adpounds employ weak C\u2014H\u22efO inter\u00adactions for inter\u00admolecular association, which are enhanced by C\u2014H\u22efCl contacts in the chlorinated products.UV irradiation of tetra\u00adhydro\u00adfuran solutions of [CpMn(CO) Limitation of the search to the fragment [(C5H5)Mn(CO)2PPh2] gave 10 hits, of which most contained unsymmetrical mono- or dinuclear diphos\u00adphanes. Relevant in the context of this study were an early determination of the structure of [(C5H5)Mn(CO)2(PPh3)] Mn(CO)2PPh2CH2Ph] by other donor ligands, particularly phosphanes, is one of the most important textbook examples for the reactivity of metal carbonyl com\u00adplexes Mn(CO)3] (I) and a slight molar excess of the phosphane in tetra\u00adhydro\u00adfuran was irradiated for 7\u2005h under argon. The colours of the solutions changed from yellow to red with concomitant gas evolution. After further stirring for 16\u2005h, the solvent was evacuated and the residue dissolved in diethyl ether (Et2O) and filtered through a plug of silica gel. The solvent was evaporated again and the residue dissolved in the minimum amount of petroleum ether. This solution was placed on top of a silica gel chromatography column and the products were eluted with a petroleum ether/Et2O (9:1 v/v) mixture. Evaporation of the eluate yielded the products as yellow powders. Recrystallization from petroleum ether (with some added Et2O) by slow evaporation in a refrigerator at 5\u2005\u00b0C yielded crystals of all three com\u00adpounds.A solution of and PCy3 in THF (120\u2005ml) was irradiated for 7\u2005h. After the usual work up (see above), a yellow solid was obtained, consisting of a 7:3 mixture of 2b and 2a. Recrystallization from petroleum ether (with some added Et2O) by slow evaporation in a refrigerator at 5\u2005\u00b0C yielded crystals. 1H NMR : \u03b4 4.63 (2H), 4.33 (2H), 2.02\u20131.07 (33H). 31P{1H} NMR : \u03b4 91.8. MS : m/z\u00a0= 490.4 (M+), 434.4 (M+ \u2212 2CO).A solution of impure [(C5H4Cl)Mn(CO)3] and dppe in THF (120\u2005ml) was irradiated for 7\u2005h. After usual work up, 3b was isolated as an orange powder. 0.05\u2005g of the starting material was recovered. Recrystallization from petroleum ether (with some added Et2O) by slow evaporation in a refrigerator at 5\u2005\u00b0C yielded crystals. 1H NMR : \u03b4 7.82\u20137.75 (4H), 7.47\u20137.35 (6H), 7.33\u20137.22 (6H), 7.19\u20137.09 (4H), 4.44 (2H), 3.56 (2H), 2.53\u20132.41 (2H), 2.36\u20132.22 (2H). 13C{1H} NMR : \u03b4 232.8 , 142.9 , 139.83\u2013138.89 (m), 133.1 , 131.4 , 129.4, 128.6, 128.1 , 97.5, 78.0, 77.9, 77.6, 30.6 . 31P{1H} NMR : \u03b4 117.6. IR : \u03bd (CO)\u00a0= 1847. MS : m/z\u00a0= 580.3 (M+), 552.3 (M+ \u2212 CO), 398.2 (C26H24P2), 183.0 (PPh2), 108.0 (PPh). HRMS (EI): m/z calculated 580.0684, found: 580.0681 (M+).A solution of in the presence of PPh3 leads to 1a and 1b in moderate yields of 40\u201360% \u2005\u00c5] is significantly longer (>20\u03c3) than the Mn1\u2014P1 bond [2.2259\u2005(6)\u2005\u00c5]. All other bond lengths are identical in the two mol\u00adecules (Table\u00a02The major difference between the two mol\u00adecules is in the relative orientation of the Mn(CO)s Table\u00a02.supporting information). Three of them involve arene C\u2014H bonds, and carbonyl atom O22 accepts two of them .There are five inter\u00admolecular C\u2014H\u22efO hydrogen bonds shorter in 1b. The most important bond parameters can be found in Table\u00a02The Mn\u2192P vector is nearly perpendicular to the C\u2014Cl bond (torsion angle C1\u2014Ct\u2014Mn\u2014P1 is 77.6\u00b0). The individual bond lengths are nearly identical to those in supporting information). The Cl atoms always bridge two different H atoms of the same symmetry-related arene ring along the a screw axis. Apparently, this inter\u00adaction enforces the orientation of this particular arene ring and generates the chirality.There is only one intra\u00admolecular C\u2014H\u22efCl hydrogen bond with a length shorter than the sum of the van der Waals radii (H16\u22efCl1). Additionally, there is one weak intra\u00admolecular and three inter\u00admolecular C\u2014H\u22efO hydrogen bonds, and one inter\u00admolecular C\u2014H\u22efCl hydrogen bond and protonation studies followed soon afterwards in very low yield. Despite long irradiation times, large amounts of the starting material could be recovered. In contrast to 1a, it was not possible to li\u00adthiate 2a with n-BuLi or t-BuLi and chlorinate the presumed inter\u00admediate lithium com\u00adpound with C2Cl6 to give 2b. It was possible, however, to obtain crystals of both com\u00adpounds suitable for X-ray diffraction.We prepared both com\u00adpounds according to Scheme\u00a012a crystallizes in the monoclinic space group P21/n, with one mol\u00adecule in the asymmetric unit Table\u00a02. The dis3 ligand and carbonyl atom O1. A packing diagram shows that these inter\u00adactions mainly join the individual mol\u00adecules in the c direction .There is one intra\u00admolecular and two inter\u00admolecular C\u2014H\u22efO hydrogen bonds involving exclusively methyl\u00adene H atoms of the PCy2b crystallizes in the monoclinic space group P21/c, with one mol\u00adecule in the asymmetric unit than in 2a and have the same lengths as in 1b. This is also true for the distance of the Mn atom from the centroid of the cyclo\u00adpenta\u00addienyl ring. More bond parameters can be found in Table\u00a02Compound it Fig.\u00a03. The Mn\u2192X inter\u00adactions involving two methyl\u00adene H atoms of the PCy3 ligand and either the Cl atom or one carbonyl O atom. Additionally, an inter\u00admolecular C\u2014H\u22efCl hydrogen bond joins glide-plane-related mol\u00adecules along the b axis .There are intra\u00admolecular C\u2014H\u22ef3a was first prepared by the photochemical reaction of cymantrene with bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)ethane (dppe) in benzene (ca 85% yield after 50\u2005h irradiation), while the same reaction in cyclo\u00adhexane produced the dppe-bridged dinuclear com\u00adplex {[(C5H5)Mn(CO)2]2[\u03bc-dppe]} , again with substantial recovery of the starting material. Some weak signals in the NMR spectra showed small amounts of other products, most likely dinuclear ones. However, the influence of prolonged reaction times on product yields and distribution was not examined. In contrast to the reactivity of 1b, it was not possible to deprotonate 3b and introduce more chlorine substituents via addition of C2Cl6. However, again it was possible to obtain crystals suitable for X-ray diffraction for both com\u00adpounds.Irradiation of THF solutions of the corresponding tricarbonyl com\u00adplexes in the presence of dppe for 7\u2005h yields 3a crystallizes in the monoclinic space group C2/c, with one mol\u00adecule in the asymmetric unit. Fig.\u00a04Compound 3 ligand or one C\u2014H group of the cyclo\u00adpenta\u00addienyl ring. The packing diagram shows that these inter\u00adactions connect the individual mol\u00adecules in the a direction .There are two inter\u00admolecular hydrogen bonds involving the carbonyl O atom and one methyl\u00adene H atom of the PCy3b crystallizes in the triclinic space group P3a. The same holds for the relative distances between manganese and the cyclo\u00adpenta\u00addienyl centroids, while the Mn\u2014CO bonds are virtually identical .There is one inter\u00admolecular C\u2014H\u22efCl hydrogen bond involving an arene H atom, which joins the individual mol\u00adecules in the 3 system, where a substantial elongation occurs. When com\u00adparing the two triads with different phosphanes, the Mn\u2014Ct (Ct des\u00adcribes the centroid of the cyclo\u00adpenta\u00addienyl ring) and Mn\u2014P distances show a slight increase in the order 3\u21921\u21922. The C\u2014O bonds follow the trend 1 \u2243 2 < 3 and the C\u2014Cl bonds follow the trend 2b < 1b \u2243 3b. The average C\u2014C bond lengths are the same within 2\u03c3 for all six com\u00adpounds. Comparison with the PPh2CH2Ph com\u00adpound GIXRIO and the ferrocenylbis\u00adphosphane chelate com\u00adpound EFUHAO shows more similarities with the PPh3 com\u00adplexes 1 than with the dppe chelates 3. The tendency of the Mn\u2014P bonds to eclipse one cyclo\u00adpenta\u00addienyl C\u2014H bond is obvious in all the com\u00adpounds. In all the chloro com\u00adpounds, the Mn\u2014P bonds avoid being eclipsed with the C\u2014Cl bond of the cyclopentadienyl ring.The introduction of a chlorine substituent in the cyclopentadienyl ring leads to a slight increase in the Mn\u2014Ct and Mn\u2014P distances for all the title phosphanes, while both the Mn\u2014CO and the C\u2014O bonds are only affected in the PCyvia the formation of C\u2014H\u22efCl hydrogen bonds.Apparently, the introduction of one chlorine substituent has only a small influence on the bond lengths, despite the relatively large effect on the spectroscopic data. Steric hindrance within the phosphanes seems to be of greater importance for the bond parameters than the differences in electronic effects. However, the presence of chlorine in the cyclo\u00adpenta\u00addienyl ring leads to additional lattice stabilization 10.1107/S2053229621009177/dv3012sup1.cifCrystal structure: contains datablock(s) compd1a, compd1b, compd2a, compd2b, compd3a, compd3b, global. DOI: 10.1107/S2053229621009177/dv3012compd1asup2.hklStructure factors: contains datablock(s) compd1a. DOI: 10.1107/S2053229621009177/dv3012compd1bsup3.hklStructure factors: contains datablock(s) compd1b. DOI: 10.1107/S2053229621009177/dv3012compd2asup4.hklStructure factors: contains datablock(s) compd2a. DOI: 10.1107/S2053229621009177/dv3012compd2bsup5.hklStructure factors: contains datablock(s) compd2b. DOI: 10.1107/S2053229621009177/dv3012compd3asup6.hklStructure factors: contains datablock(s) compd3a. DOI: 10.1107/S2053229621009177/dv3012compd3bsup7.hklStructure factors: contains datablock(s) compd3b. DOI: 10.1107/S2053229621009177/dv3012sup8.pdfAdditional figures and table. DOI: 2107491, 2107490, 2107489, 2107488, 2107487, 2107486CCDC references:"} +{"text": "In this work, practically important relationships between Eint and electron density, its Laplacian, curvature, potential, kinetic, and total energy densities at the bond critical point as well as bond length were derived for the structures of the [Z\u2013I\u00b7\u00b7\u00b7Hal]\u2212 and [Z\u2013Hal\u00b7\u00b7\u00b7I]\u2212 types bearing halogen bonds and involving iodine as interacting atom(s) . The mean absolute deviations for the correlations found were 2.06\u20134.76 kcal/mol.Bond energy is the main characteristic of chemical bonds in general and of non-covalent interactions in particular. Simple methods of express estimates of the interaction energy, E The halogen bond is one of the most important types of non-covalent interactions being second only to hydrogen bonds in its significance. According to the IUPAC definition, \u201ca halogen bond occurs when there is evidence of a net attractive interaction between an electrophilic region associated with a halogen atom in a molecular entity and a nucleophilic region in another, or the same, molecular entity\u201d . Weak inThe supramolecular and cluster chemistry of halide anions currently attracts much attention ,27,28,29b) is a difficult task. Experimental methods are usually associated with complex technical procedures and can be applied to a very limited number of structures. The direct theoretical calculations of Eb for the A\u00b7\u00b7\u00b7B bond as the energy difference between the structure A\u00b7\u00b7\u00b7B and the isolated molecules A and B may be successfully applied to intermolecular non-covalent interactions in the gas phase. However, in the condensed phase, molecules are usually bound with each other by a network of several non-covalent interactions, and an adequate fragmentation which affects only the bond of interest is often impossible. In such a situation, an approximation of Eb through other parameters easily accessible from experiment becomes very important \u2212 and [(A)nZ\u2013Y\u00b7\u00b7\u00b7I]\u2212 types were calculated. The correlations between Eint and the electron density (\u03c1b), its Laplacian (\u22072\u03c1b), the curvature of \u03c1(r) which is parallel to the bond path direction (positive) , the potential, kinetic, and total energy densities at BCP, and the halogen bond length (dY\u00b7\u00b7\u00b7X) were established.Recently, the author started a project to establish practically useful correlations between the interaction energy and properties which could be easily determined from experiment for halogen bonds of various types, including those formed by a halide anion. In the previous work , such renZ\u2013Y\u00b7\u00b7\u00b7X]\u2212 which include different types of second-order atoms Z and third-order groups A, otherwise, the relationships found may suffer a significant overfitting. Therefore, a large statistically significant set of 412 structures bearing twelve different types of the Z atom was used in this work.Two points should be mentioned here. First, the interactions with the iodine atom as a halogen bond donor are the most important among all halogen bonds since the existence of a prominent \u03c3-hole at the iodine atom provides particularly high stability of these interactions. Second, reliable relationships can be obtained only for sufficiently large sets of the structures [(A)Full geometry optimization of the main set of structures was carried out at the density functional theory (DFT) level by using the M06-2X functional with thenZ\u2013Y\u00b7\u00b7\u00b7X]\u2212 structures [int(Vb) correlations for the [(A)nZ\u2013Y\u00b7\u00b7\u00b7X]\u2212 structures obtained at the M06-2X level of theory have similar parameters as those for the MP4, CCSD, and CCSD(T) methods. On the other hand, it was found \u2212 were calculated using Equation (1):nZ\u2013Y fragments were unrelaxed and corresponded to those in [(A)nZ\u2013Y\u00b7\u00b7\u00b7X]\u2212. The topological analysis of the electron density distribution was performed with the help of the Atoms in Molecules (AIM) method developed by Bader \u2212 and [(A)nZ\u2013Y\u00b7\u00b7\u00b7I]\u2212 types were selected for the calculations. Among them, 70 structures were in each series [(A)nZ\u2013I\u00b7\u00b7\u00b7X]\u2212 with X = F, Cl, or Br and [(A)nZ\u2013Br\u00b7\u00b7\u00b7I]\u2212, 65 structures were in the [(A)nZ\u2013I\u00b7\u00b7\u00b7I]\u2212 series, and 67 structures were in the [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 series. For Z = C and N, different orbital hybridizations of these atoms were considered. For Z = S and P, different oxidation states of these atoms were considered. The groups A vary from the electron donor to the electron acceptor ones providing a broad span of interaction energies for the halogen bond. The complete list of the calculated structures is given in Structures of the [(A)nZ\u2013I\u00b7\u00b7\u00b7F]\u2212 type O2S\u2013I\u2026F]\u2212, [F3Si\u2013I\u2026F]\u2212, [p-H2N-C6H4-Br\u2013I\u2026F]\u2212, [Me3Si\u2013I\u2026F]\u2212, [MeO\u2013I\u2026F]\u2212, [(Me)O2S\u2013I\u2026F]\u2212, [Ph\u2013I\u2026F]\u2212, and [(p-Me-C6H4)2N\u2013I\u2026F]\u2212), which provide the Eint span from \u20139 to \u201373 kcal/mol.The M06-2X/ADZP\u2013DKH method was validated toward the reproduction of the experimental electron densities for the set of eleven structures bearing halogen bonds of different types and for which the experimental electron densities are known. Performance of the M06-2X and PBE0-D3BJ functionals was also compared for the set of ten structures of the \u2212, HO\u2013H\u00b7\u00b7\u00b7Cl\u2212, HO\u2013H\u00b7\u00b7\u00b7I\u2212, H3C\u2013H\u00b7\u00b7\u00b7I\u2212, and [I\u2013I\u2013I]\u2212). As can be seen in \u2212. The experimental error is in the range of 0.1\u20131.6 kcal/mol, and theoretical M06-2X values fall within the 3\u03c3 interval for four of six structures. The interaction energies calculated at the CCSD and CCSD(T) levels were strongly underestimated when the CP correction was applied. The CCSD method performed worse than M06-2X for H3C\u2013H\u00b7\u00b7\u00b7F\u2212 and HO\u2013H\u00b7\u00b7\u00b7Cl\u2212 even with the aug-cc-pVTZ basis set, and only the CCSD(T) level with the triple-zeta basis set and without the CP correction provided better results than the DFT approach . Thus, considering that typical MAD values for the Eint property relationships found for the structures [(A)nZ\u2013Hal1\u00b7\u00b7\u00b7Hal2]\u2212 are 2\u20134 kcal/mol . cations . Here, tructures . The calty 1.07, . The PBEint(Vb) relationships obtained for the small set of ten structures of the [(A)nZ\u2013I\u00b7\u00b7\u00b7F]\u2212 type were similar for both M06-2X and PBE0-D3BJ functionals and it was 3.96 kcal/mol at Vb = 100 kcal/(mol\u2022bohr3). These deviations were lower than MAD for the whole set of seventy [(A)nZ\u2013I\u00b7\u00b7\u00b7F]\u2212 structures . All these results indicate that the relationships found in this work at the M06-2X/ADZP\u2013DKH level are reliable for the determination of Eint using experimental electron density properties.Correspondingly, the Ectionals . The devnZ\u2013I\u00b7\u00b7\u00b7X]\u2212 and [(A)nZ\u2013Y\u00b7\u00b7\u00b7I]\u2212 types and the corresponding Eint property relationships were discussed (in this and the following sections). The calculated BSSE-corrected interaction energies for the halogen bonds in these structures varied between 2.53 and \u221288.43 kcal/mol. These bonds were the strongest for the [(A)nZ\u2013I\u00b7\u00b7\u00b7F\u2212] series. The highest dispersion of Eint was found for the [(A)nZ\u2013I\u00b7\u00b7\u00b7F]\u2212 and [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 series. The halogen bonds in the series [(A)nZ\u2013I\u00b7\u00b7\u00b7Cl]\u2212, [(A)nZ\u2013I\u00b7\u00b7\u00b7Br]\u2212, and [(A)nZ\u2013I\u00b7\u00b7\u00b7I]\u2212 have similar strengths and dispersion.After validation of the computational method, the interaction energies calculated for 412 structures of the [(A)int for the hydrogen bonds X\u2013H\u2026O and FH\u2026FR [nZ\u2013Y\u00b7\u00b7\u00b7X]\u2212 bearing halide anions [int and Vb were found for all series except [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 (int(Gb) relationship was linear only for the [(A)nZ\u2013I\u00b7\u00b7\u00b7F]\u2212 series and it was approximated by a binominal function for [(A)nZ\u2013I\u00b7\u00b7\u00b7X]\u2212 and [(A)nZ\u2013Br\u00b7\u00b7\u00b7I]\u2212. Finally, the Eint(\u03c1b) dependence was quadratic for all series.These types of relationships were recommended for the predictions of End FH\u2026FR ,83 and fe anions . Here, tCl\u00b7\u00b7\u00b7I]\u2212 . The EinnZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 series (int(Vb), Eint(Gb), or Eint(\u03c1b) function. Instead, the horizontal branch is clearly visible in the range of Eint of 10\u201320 kcal/mol. The interaction energy could be reasonably approximated by polynomial fourth-order functions, but such an approximation is usually associated with significant overfitting and cannot be recommended for practical use. The reasons of this peculiar situation are discussed below.An interesting relationship distinct from all other bonds with halide anions was obtained for the [(A)\u2212 series . These snZ\u2013Y\u00b7\u00b7\u00b7X]\u2212 in this and previous \u2212. Third, the slope of the relationships is strongly affected by the nature of the Y and X atoms. For instance, for the Vb estimator, the lowest slope (0.69\u20130.84) and the highest negative intercept (\u22123.68 \u00f7 \u22126.50) were found for X = F. The highest slopes were detected for [(A)nZ\u2013I\u00b7\u00b7\u00b7I]\u2212 (2.34) as well as for the [(A)nZ\u2013Y\u00b7\u00b7\u00b7X]\u2212 series (1.55\u20132.12). The series with (X or/and Y) = Cl but (X and Y) \u2260 F exhibited an intermediate slope (1.26\u20131.46). Such a behavior can be qualitatively interpreted by the different properties of the halogen atoms forming the Y\u00b7\u00b7\u00b7X bond. First of all, the electronegativity of the X atom decreases along the series F > Cl > Br > I. There is a clear trend between the slope of the Eint(Vb) dependence and the difference of the electronegativities of X and Y nZ group on the Y\u00b7\u00b7\u00b7X bond are more pronounced if the electron shells of X are more diffuse, i.e., from X = F to I. Since the higher dispersion contribution and more diffuse electron shells are associated with lower concentration of electron or energy density at BCP, Eint values are higher for most electron acceptor groups (A)nZ at a given value of Eint in the case of heavier atom X.Analysis of the relationships obtained for the structures \u2212 + [(A)nZ\u2013I\u00b7\u00b7\u00b7Br]\u2212 + [(A)nZ\u2013Br\u00b7\u00b7\u00b7I]\u2212, (ii) [(A)nZ\u2013I\u00b7\u00b7\u00b7Cl]\u2212 + [(A)nZ\u2013Cl\u00b7\u00b7\u00b7Cl]\u2212 + [(A)nZ\u2013Br\u00b7\u00b7\u00b7Cl]\u2212 + [(A)nZ\u2013Cl\u00b7\u00b7\u00b7Br]\u2212, and (iii) [(A)nZ\u2013I\u00b7\u00b7\u00b7F]\u2212 + [(A)nZ\u2013Br\u00b7\u00b7\u00b7F]\u2212 + [(A)nZ\u2013Cl\u00b7\u00b7\u00b7F]\u2212 groups. Each of them may be approximated by a linear function . The Eint(Gb) and Eint(\u03c1b) dependences are characterized by a higher dispersion between series. They are more sensitive to the nature of the Y and X atoms and, therefore, such a grouping is not efficient for those relationships.Fourth, some series form distinct groups which can be approximated by a single relationship of a reasonable quality. There are three such groups when considering the EnZ\u2013Y\u00b7\u00b7\u00b7X]\u2212 structures were significantly different from those obtained previously for the hydrogen bonds X\u2013H\u2026O and FH\u2026FR \u2212 (this work) and [(A)nZ\u2013Br\u00b7\u00b7\u00b7F]\u2212 [int(\u22072\u03c1b) functions for other series are not well-defined nZ\u2013I\u00b7\u00b7\u00b7F]\u2212, [(A)nZ\u2013I\u00b7\u00b7\u00b7Cl]\u2212, and [(A)nZ\u2013I\u00b7\u00b7\u00b7Br]\u2212 relationship for the first of these series is linear and of a lower quality compared to the Eint(Vb), Eint(Gb), and Eint(\u03c1b) functions. The last two series are very well approximated by two exponential functions . For the series [(A)nZ\u2013I\u00b7\u00b7\u00b7I]\u2212 and [(A)nZ\u2013Br\u00b7\u00b7\u00b7I]\u2212, the \u03bb||,b parameter is not sensitive to determine Eint at high values fittings at lower Eint are also not reasonable. Finally, the Eint function is not well-defined for [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212.The curvature of the electron density distribution can serve for the estimates of EI\u00b7\u00b7\u00b7Br]\u2212 . The Eincal/mol) . The Einint depends on the nature of the Y and X atoms. It cannot be used for the [(A)nZ\u2013I\u00b7\u00b7\u00b7F]\u2212 series due to high dispersion of the Eint(Hb) function at lower Eint and its low sensitivity at higher Eint (int(Hb) relationship is not sensitive for the weakest interactions with Eint < 10 kcal/mol. However, for stronger interactions, good linear dependences were observed , except for the [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 series which was approximated by a quadratic function nZ\u2013I\u00b7\u00b7\u00b7I]\u2212, [(A)nZ\u2013Br\u00b7\u00b7\u00b7I]\u2212, [(A)nZ\u2013Cl\u00b7\u00b7\u00b7Cl]\u2212, [(A)nZ\u2013Cl\u00b7\u00b7\u00b7Br]\u2212, and [(A)nZ\u2013Br\u00b7\u00b7\u00b7Br]\u2212) are described together by a single linear function of the reasonable quality . Six sercal/mol) .nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 demonstrate reasonable exponential relationships between interaction energy and the length of the halogen bond nZ\u2013I\u00b7\u00b7\u00b7Br]\u2013 + [(A)nZ\u2013Br\u00b7\u00b7\u00b7I]\u2212 and [(A)nZ\u2013Br\u00b7\u00b7\u00b7Cl]\u2212 + [(A)nZ\u2013Cl\u00b7\u00b7\u00b7Br]\u2212. For other series, the fitting parameters are quite different from each other. Similarly to the other estimators, the Eint(dY\u2026X) dependence is complex for the [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 series. There is a horizontal branch for the interval of Eint 5\u201315 kcal/mol. Meanwhile, in this case, the dependence may be quite reasonably approximated by a two-exponential function .All series except for [(A)gen bond . The quaint\u2013property relationships for the [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 series have a peculiar character. The more detailed analysis shows that such behavior is due to the tremendous effect of the second-order atom Z and the third-order groups A. First, the Eint values are low and do not exceed 14 kcal/mol when Z = C, P, Si, B, or H and those with Z = O, N, or S have a significant span of Eint .As was mentioned above, the E B, or H . The strint(Vb) trend slope for the combined series [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 (Z = Si + B) is 6.75 while the slope of the linear correlation for the series [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 is 1.43 are not linear but exponential. The tangent slopes at the lowest energy points are 9.2 and 8.7.Second, the E is 1.43 . DependenN\u2013Cl\u00b7\u00b7\u00b7I]\u2212 structures could be divided into two distinct groups and those with a stronger Cl\u00b7\u00b7\u00b7I interaction imine and nitro compounds, Group II). The trend slope for these two groups is similar (2.1) but the intercept is very different . The different behavior of these two groups is apparently associated with a different nature of the (A)nN part of the molecule. In Group I, (A)nN exhibits either electron donor or weak electron acceptor properties. In Group II, (A)nN is a strong electron acceptor. Furthermore, the structures of Group II have more extended conjugation systems compared to those of Group I. All these relationships considered together provide such a peculiar trend for the whole [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 series. Interestingly, this effect of Z and A is much smaller for other series making possible the existence of linear correlations. Similar behavior was also found for the Gb and \u03c1b estimators.Third, the \u2212 series. However, for the more narrow series \u2212, [(A)nC\u2013Cl\u00b7\u00b7\u00b7I]\u2212, and [(A)nP\u2013Cl\u00b7\u00b7\u00b7I]\u2212, these relationships are of a good quality and they may be useful for practical applications. The d(Y\u2026X) estimator is much better for the description of the [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212 series because it permits the reasonable approximation of Eint by a single two-exponent function.The great effect of the Z atom and A group on Eint for the condensed phases is a much more difficult task than that for the gas phase. Both periodic solid state and cluster approaches often do not allow direct estimates of Eint as an energy difference because fragments may have multiple mutual weak intermolecular interactions. When it is possible, the adequate computational model is usually exceedingly large. For instance, accurate calculations of Eint for the A\u00b7\u00b7\u00b7B bond in a solid-state structure within the cluster approach requires explicit consideration of all intermolecular interactions which molecules A and B are involved in. An example of an adequate cluster for the direct estimates of Eint between the simple fragments I\u2013C4F8\u2013I and Cl\u2212 in the X-ray structure ACIPOU is shown in 4F8\u2013I\u00b7\u00b7\u00b7Cl\u2212 surrounded by all molecules which form short contacts with the I\u2013C4F8\u2013I or Cl\u2212 fragments).The estimate of EConsidering this situation, the practice of utilization of small bimolecular clusters which include only two molecules forming a contact of interest becomes increasingly popular. In other words, there are multiple attempts to approximate the properties of solid-state structures by gas phase calculations.int calculated for the I\u2013CF2CF2\u2013I\u00b7\u00b7\u00b7Cl\u2212 halogen bond in the X-ray structure ACIPIO using the bimolecular approach is 20.07 kcal/mol. However, when four molecules are considered .There are two points which invalidate this practice. First, such a bimolecular model ignores other intermolecular interactions in which both considered molecules participate. These secondary interactions may significantly affect the energy of the contact under study. For instance, Eint has little physical meaning.The second point is that geometry of bimolecular clusters is usually not optimized but taken from the X-ray structures since optimization often leads to the collapse of the structure compared to X-ray. In such calculations, the geometry considered is not equilibrium and does not belong to a minimum energy path for either adiabatic or vertical bond cleavage, and, hence, the resulting Eint in the condensed phase is free from the difficulties discussed above since it requires a single parameter directly obtained from experiment.Application of the correlations established in this work for the rapid estimates of EnZ\u2013I\u00b7\u00b7\u00b7Hal]\u2212 and [(A)nZ\u2013Hal\u00b7\u00b7\u00b7I]\u2212 bearing the iodine atom . Thus, together with the results published previously \u2212, those with X = F demonstrate quite poor approximations of Eint with MAD of 3.29\u20134.76 kcal/mol. For other structures, except [(A)nZ\u2013Cl\u00b7\u00b7\u00b7I]\u2212, more reasonable correlations were obtained . Meanwhile, the approximations of Eint for halogen bonds formed by halide anions are worse than those obtained for homohalogen bonds formed by two neutral fragments \u2212 type. In fact, only two parameters were found to be able to adequately estimate Eint for this bond, i.e., Hb and dY\u2026X.Second, the most difficult case is the structures of the [(A)int for all structures [(A)nZ\u2013Y\u00b7\u00b7\u00b7X]\u2212. For the electron density-based properties, the highest slopes (linear correlations) or curvatures (non-linear correlations) were found for the bonds formed by heavy halogens (I and Br) while the lowest slopes and curvatures were observed for X = F.Third, there is no unique equation to approximate EFourth, the correlations derived for halogen bonds with halide anions are very different from those established for some hydrogen bonds and halogen bonds formed by two neutral fragments . This anb, Vb, Gb, and dY\u00b7\u00b7\u00b7X estimators behave similarly. In contrast, the \u22072\u03c1b, \u03bb||,b, and Hb parameters have limited significance as Eint estimators and they may be used only for some series or a certain range of the Eint values.Fifth, the \u03c1int of the structures [(A)nZ\u2013Y\u00b7\u00b7\u00b7I]\u2212 and [(A)nZ\u2013I\u00b7\u00b7\u00b7X]\u2212 are given in The relationships recommended for practical use to estimate E"} +{"text": "The distorted KO6 octa\u00adhedra share edges, resulting in chains running in the [010] direction.The title compound, C E)-2-{4-[3-(thio\u00adphen-3-yl)acrylo\u00adyl]phen\u00adoxy}acetic acid are described. Crystallization from an ethanol\u2013water mixture resulted in the title compound, C30H23KO8S2 or [K(C15H11O4S)(C15H12O4S)]n, containing one mol\u00adecule of the acid and one mol\u00adecule of the potassium salt in the asymmetric unit. Both mol\u00adecules share the H atom between their carboxyl groups and a potassium ion. The C=C bonds display an E configuration. The thio\u00adphene and phenyl rings in the two mol\u00adecules are inclined by 43.3\u2005(2) and 22.7\u2005(2)\u00b0. The potassium ion is octa\u00adhedrally coordinated by six O atoms. This distorted octa\u00adhedron shares on opposite sides two oxygen atoms with inversion-related octa\u00adhedra, resulting in chains of octa\u00adhedra running in the [010] direction, which form ladder-like chains by C\u2014H\u22ef\u03c0 inter\u00adactions. A Hirshfeld surface analysis indicates that the highest contributions to the surface contacts arise from inter\u00adactions in which H atoms are involved, with the most important contribution being from H\u22efH (31.6 and 31.9% for the two mol\u00adecules) inter\u00adactions.The synthesis and spectroscopic data of ( This compound is considered to be a good monomer for the synthesis of water-soluble polythio\u00adphene-based conjugated polyelectrolytes. A single-crystal structure determination indicates that after crystallization, crystals were obtained containing one mol\u00adecule of the acid and one mol\u00adecule of the potassium salt in the asymmetric unit.\u00b0 for mol\u00adecule B. The C=C bonds display an E configuration, resulting in short intra\u00admolecular C6\u2014H6\u22efO9 and C26\u2014H26\u22efO29 inter\u00adactions , K41\u22efO36ii = 3.347\u2005(3)\u2005\u00c5, symmetry codes: (ii) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01, (iv) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01].Potassium ion K41 is octa\u00adhedrally coordinated by six O atoms with K\u2014O distances between 2.672\u2005(2) and 2.906\u2005(3)\u2005\u00c5 Fig.\u00a03 and an oii, at the other side O20i and O20iv; symmetry codes: (i) x\u00a0+\u00a01, y, z, (ii) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01, (iv) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01] with inversion-related octa\u00adhedra and 4.8353\u2005(14)\u2005\u00c5 .In the crystal packing, the potassium ion K41 inter\u00adacts with six mol\u00adecules of which two occur in the carb\u00adoxy\u00adlic acid form Fig.\u00a04. The disa Figs. 3 and 5 \u25b8.Cg1v contacts .Despite the presence of many aromatic rings, the crystal packing of the title compound does not show \u03c0any \u2013\u03c0 inter\u00adactions. The shortest centroid\u2013centroid distance is 4.735\u2005(3)\u2005\u00c5 between thio\u00adphene rings S1/C2\u2013C5 and S21/C22\u2013C25 with an angle between the rings of 52.3\u2005(3)\u00b0. However, C\u2014H\u22ef\u03c0 inter\u00adactions are present and give rise to a ladder-like chain also running in the [010] direction Table\u00a01. In addiThe packing does not show any residual solvent-accessible voids.et al., 2007CrystalExplorer . The sharp tips at de + di \u2243 1.4\u2005\u00c5 arise from the O19\u2014H19\u22efO39 hydrogen bond. The principal contribution to the Hirshfeld surfaces involves H\u22efH contacts at 31.6 and 31.9% for mol\u00adecules A and B, respectively. These are followed by C\u22efH/H\u22efC (21.1 and 20.0%) O\u22efH/H\u22efO (17.4 and 17.3%) and S\u22efH/H\u22efS (8.8 and 9.9%) contacts.The Hirshfeld surface analysis \u2014p-C6H4\u2014R2 gave 619 hits (with C atoms double-bond acyclic). For only 33 cases (5.3%), the double bond has the Z configuration . The histogram of the dihedral angle between the planes of the double bond and the phenyl ring shows values between 0.0 and 86.2\u00b0 . A search with thio\u00adphene as R1 resulted in only four hits . Only one structure was found for which R2 is the same as in the title compound -2-{4-[3-(thio\u00adphen-3-yl)acrylo\u00adyl]phen\u00adoxy}acetic acid, is given in Fig.\u00a09The synthetic pathway to synthesize the target compound, (A mixture of ethyl 2-(4-acetyl\u00adphen\u00adoxy)acetate (5\u2005mmol), 3-thio\u00adphene\u00adcarbaldehyde (5\u2005mmol) and 50\u2005mL of ethanol was stirred in ice-cold water for 20 minutes. Then, 5\u2005mL of 50% KOH solution was added dropwise to the reaction mixture, which was then stirred continuously for 5\u2005h. At the end of the reaction, water was added to the reaction mixture and stirring was continued until all solids in the mixture were dissolved. Concentrated HCl was slowly added to the obtained solution until the solution changed from brown to yellow. The solution was then heated until crystals appeared. The solid then began to crystallize when the solution temperature started to decrease. The crystallized solid was filtered off, washed thoroughly with water and recrystallized from an ethanol\u2013water mixture to give 2-{4-[3-(thio\u00adphen-3-yl)acrylo\u00adyl]phen\u00adoxy}acetic acid (yield 62%) in the form of pale-yellow crystals (m.p. 455\u2005K).\u22121): 1017, 980 (=C\u2014H bend), 1597 (C=C), 1659 (C=O), 3457 .IR , J (Hz)]: 7.60 , 7.42 , 7.38 , 7.81 , 7.34 , 8.03 , 7.02 , 4.77 .13C NMR : 121.81 (C2), 128.75 (C3), 127.01 (C4), 125.41 (C5), 132.67 (C6), 130.87 (C7), 171.85 (C8), 169.73 (C9), 138.39 (C10 and C14), 137.96 (C11 and C13), 114.65 (C12), 64.68 (C15), 189.09 (C16). Calculation for C15H11O4S: M = 287 au.Uiso(H) value of 1.5Ueq of the parent atom O19. The other H atoms were placed in idealized positions and included as riding contributions with an Uiso(H) values of 1.2Ueq of the parent atom, with C\u2014H distances of 0.93 (aromatic) and 0.97\u2005\u00c5 (CH2). In the final cycles of refinement, 12 outliers with |error/e.s.d.| > 5.0 were omitted.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021004801/ey2007sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021004801/ey2007Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021004801/ey2007Isup3.cmlSupporting information file. DOI: 2082049CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The natural convection boundary layer flow of a viscous incompressible fluid with temperature dependent viscosity and thermal conductivity in the presence of exothermic catalytic chemical reaction along a curved surface has been investigated. The governing non dimensional form of equations is solved numerically by using finite difference scheme. The numerical results of velocity profile, temperature distribution and mass concentration as well as for skin friction, heat transfer rate and mass transfer rate are presented graphically and in tabular form for various values of dimensionless parameters those are generated in flow model during dimensionalization. From the obtained results, it is concluded that the exothermic catalytic chemical reactions is associated with temperature dependent viscosity and thermal conductivity. Further, it is concluded that the body shape parameter also plays an important quantitative role for change in velocity profile, temperature field and mass concentration behavior in the presence of exothermic catalytic chemical reaction. Merkin and Chaudhary .At this time, please address the following queries:Please clarify the sources of funding for your study. List the grants or organizations that supported your study, including funding received from your institution.State what role the funders took in the study. If the funders had no role in your study, please state: \u201cThe funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201dIf any authors received a salary from any of your funders, please state which authors and which funders.If you did not receive any funding for this study, please state: \u201cThe authors received no specific funding for this work.\u201dPlease include your amended statements within your cover letter; we will change the online submission form on your behalf.Reviewers' comments:5. Review Comments to the AuthorReviewer #1:\u00a0The report presents the natural convection boundary layer flow of a viscous incompressible fluid with temperature-dependent viscosity and temperature-dependent thermal conductivity in the presence of exothermic catalytic\u00a0chemical reaction along a curved surface. I recommend major revision due to the following facts.1. In Eq. (3), the authors added (u^2/2x)n to the convective acceleration term.Comment: What does the new term means? How did you derive it?2. After Eq. (6), it was written that n = P(x) + Q(x). Whereas is the wall temperature function and the is the body shape function.Comment: The authors refer Readers to check \"Pop, I. and Takhar, H. S. Free convection from a curved surface. Journal of Applied Mathematicsand Mechanics, 73 (1993), pp. 534 -539\" for details.Comment: This is not acceptable. Update the research methodology with the derivation of such momentum equation.3. Research question is needed to harmonize the contribution of the report to the body of knowledge.Comment: Research questions are needed at the end of the introduction section. Research questions are needed. Note that the results in this report are typical answers to unknown questions. This is true because the manuscript provides some powerful answers to unknown questions. Note that the research questions must connect the title to the analysis of results, and conclusion. This would guide authors not to generate many results that are not consistent to provide insight. The author should update the manuscript with appropriate and relevant research questions at the end of the introduction section. This would guide the author to structure logical analysis of results. Logical questions are expected. This would help readers to link what is known in the literature with the novelty of this study.4. The similarity variables presented as Eq. (7) are dimensional. The unit of the first, u, is m^0.5 while that of v is m^{-1/4}, and that of y is also dimensional. Hence, the variables used to non-dimenzionalize the governing equation are not appropriate.Reviewer #2:\u00a0Recommendation: Minor revisionAuthors should revise the manuscript according to the following comments.\u2022 Nomenclature is must.\u2022 Some grammatical and typo mistakes are found in the manuscript.\u2022 The governing differential equations describing the flow are non-linear. Is the solution obtained unique?\u2022 Specify the applications of considered physical model or geometry.\u2022 Introduction section should be made more concise to show previous work in the field. At present a lot of related researches are stated in the introduction. However, not sufficient analysis is presented. The authors should ask themselves: what are the problems with the presented researches? Why is the recent work needed? Hope these can improve the present work by following past articles.\u2022 Effect of Joule heating on MHD non\u2010Newtonian fluid flow past an exponentially stretching curved surface\u2022 Magnetohydrodynamic Cattaneo-Christov flow past a cone and a wedge with variable heat source/sink\u2022 Heat and mass transfer in MHD Casson nanofluid flow past a stretching sheet with thermophoresis and Brownian motion\u2022 Effect of asymmetrical heat rise/fall on the film flow of magnetohydrodynamic hybrid ferrofluid\u2022 Simultaneous solutions for first order and second order slips on micropolar fluid flow across a convective surface in the presence of Lorentz force and variable heat source/sink\u2022 Effect of thermal radiation on MHD Casson fluid flow over an exponentially stretching curved sheet\u2022 Influence of non-uniform heat source/sink on the three-dimensional magnetohydrodynamic Carreau fluid flow past a stretching surface with modified Fourier\u2019s law\u2022 MHD Carreau Fluid Flow Past a Melting Surface with Cattaneo-Christov Heat Flux\u2022 Physical aspects on unsteady MHD\u2010free convective stagnation point flow of micropolar fluid over a stretching surface\u2022 Influence of viscous dissipation on MHD flow of micropolar fluid over a slendering stretching surface with modified heat flux model\u2022 A non\u2010Fourier heat flux model for magnetohydrodynamic micropolar liquid flow across a coagulated sheetNote: I want to see the revised version of the manuscript.https://pacev2.apexcovantage.com/. 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Please note that Supporting Information files do not need this step.While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool,\u00a0 13 Apr 2021Response to Reviewers Comments (PONE-D-21-04681)Effects of Temperature Dependent Viscosity and Thermal Conductivity on Natural Convection Flow along a Curved Surface in the Presence of Exothermic Catalytic Chemical ReactionReviewer #1: The report presents the natural convection boundary layer flow of a viscous incompressible fluid with temperature-dependent viscosity and temperature-dependent thermal conductivity in the presence of exothermic catalytic chemical reaction along a curved surface. I recommend major revision due to the following facts.In Eq. (3), the authors added (u^2/2x)n to the convective acceleration term.Comment: What does the new term means? How did you derive it.Response: During dimensionalization of the momentum equation we have equation of the form u \u2202u/\u2202x+v \u2202u/\u2202y+u^2/2 1/g_x (dg_x)/dx=\u03b3_\u03bc (\u2202\u03b8/\u2202y)(\u2202u/\u2202y) +(1+\u03b3_\u03bc \u03b8)((\u2202^2 u)/(\u2202y^2 ))+\u03b8+\u03d5,Here, as 1/x \u00ae (P(x \u00ae)+Q(x \u00ae))=1/g_x (dg_x)/(dx \u00ae )and P(x \u00ae )+Q(x \u00ae )=n and thus 1/x \u00ae (n)=1/g_x (dg_x)/(dx \u00ae )u \u2202u/\u2202x+v \u2202u/\u2202y+u^2/2x n=\u03b3_\u03bc (\u2202\u03b8/\u2202y)(\u2202u/\u2202y) +(1+\u03b3_\u03bc \u03b8)((\u2202^2 u)/(\u2202y^2 ))+\u03b8+\u03d5, 2. After Eq. (6), it was written that n = P(x) + Q(x). Whereas is the wall temperature function and the is the body shape function.Comment: The authors refer Readers to check \"Pop, I. and Takhar, H. S. Free convection from a curved surface. Journal of Applied Mathematics and Mechanics, 73 (1993), pp. 534 -539\" for details.Comment: This is not acceptable. Update the research methodology with the derivation of such momentum equation.Response: The detail derivation of n=P(x)+Q(x) is given in below steps:Since we know that d/dx (lnx)=1/xand d(lnx)=1/x dxwhich implies that 1/(d(lnx))=x/dx. (1) Also consider d/dx (ln g_x )=1/g_x (dg_x)/dxwhich implies d(ln g_x )=1/g_x dg_x. (2) Now combining the above two equations (1) and (2), we have (d lng_x)/(d lnx)=x/g_x (dg_x)/dx.Now using the non-dimensionless variables, we may write the above equations as(d lng_x)/(d lnx \u00ael)=(x \u00ael)/g_x (dg_x)/(dx \u00ael)which implies that(d lng_x)/(d lnx \u00ael)=x \u00ae/g_x (dg_x)/(dx \u00ae )and thus we have 1/x \u00ae (d lng_x)/(d lnx \u00ael)=1/g_x (dg_x)/(dx \u00ae ) (3)Also, we consider d/dx (lnT_w )=1/T_w (dT_w)/dx=0which implies that d lnT_w=0and thus, we can write (d lnT_w)/(d lnx \u00ael)=0 (4)Now from equation (3) and (4), we have 1/x \u00ae ((d lnT_w)/(d lnx \u00ae l)+(d lng_x)/(d lnx \u00ae l))=1/g_x (dg_x)/(dx \u00ae )which can also be written as 1/x \u00ae (P(x \u00ae)+Q(x \u00ae))=1/g_x (dg_x)/(dx \u00ae ) (5)Here, P(x \u00ae), the wall temperature function and Q(x \u00ae) the body shape function, are defined as P(x \u00ae )=(d lnT_w)/(d lnx \u00ae l), Q(x \u00ae )=(d lng_x)/(d lnx \u00ae l) . (6) 3. Research question is needed to harmonize the contribution of the report to the body of knowledge.Comment: Research questions are needed at the end of the introduction section. Research questions are needed. Note that the results in this report are typical answers to unknown questions. This is true because the manuscript provides some powerful answers to unknown questions. Note that the research questions must connect the title to the analysis of results, and conclusion. This would guide authors not to generate many results that are not consistent to provide insight. The author should update the manuscript with appropriate and relevant research questions at the end of the introduction section. This would guide the author to structure logical analysis of results. Logical questions are expected. This would help readers to link what is known in the literature with the novelty of this study.Response: The overall objectives of this research are to develop the mathematical model to study and compare the different modes of coupling the exothermic and catalytic chemical reaction via momentum, energy and mass concentration equation. Model predictions are used to assess the effects of different parameters on conversion of exothermic catalytic chemical reaction at heated curve generated by tangential component of acceleration due to gravity. The proposed model is used as the guidelines for the selection of a suitable coupling to achieve the desired applications. A numerical technique Finite Difference Method in conjunction with primitive variable formulation is used to investigate the coupling of exothermic reaction with catalyst particles. Furthermore, parametric effects of heated wall and mass concentration along the curved surface are studied and highlighted graphically and as well as in tabular form. 4. The similarity variables presented as Eq. (7) are dimensional. The unit of the first, u, is m^0.5 while that of v is m^{-1/4}, and that of y is also dimensional. Hence, the variables used to non-dimenzionalize the governing equation are not appropriate.Response: In the section of numerical analysis, we have used primitive variable formulation given as below: u=x^(1\u20442) U,v=x^((-1)\u20444) V, x=X,y=x^(1\u20444) Y, \u03b8=\u0398(X.Y), C=\u03a6. It is pertinent to mention that primitive variable are dimensionless, we use this transformation to get primitive form of partial differential equations, please see red terms in each equation After using the aforementioned transformations, we have the following form of system of equations:U/2+X \u2202U/\u2202X-Y/4 \u2202U/\u2202Y+\u2202V/\u2202Y=0,[1/2+n/2] U^2+XU \u2202U/\u2202X+(V-YU/4) \u2202U/\u2202Y=\u03b3_\u03bc \u2202\u0398/\u2202Y \u2202U/\u2202Y+(1+\u03b3_\u03bc \u0398) (\u2202^2 U)/(\u2202Y^2 )+\u0398+\u03a6,XU \u2202\u0398/\u2202X+(V-YU/4) \u2202\u0398/\u2202Y=1/Pr [\u03be(\u2202\u0398/\u2202Y)^2+(1+\u03be\u0398) (\u2202^2 \u0398)/(\u2202Y^2 )]+\u03b2\u03bb^2 X^(1\u20442) (1+n\u03b3\u0398)exp((-E)/(1+\u03b3\u03b8))\u0398,XU \u2202\u03a6/\u2202X+(V-YU/4) \u2202\u03a6/\u2202Y=1/Sc (\u2202^2 \u03a6)/(\u2202Y^2 )+\u03bb^2 X^(1\u20442) (1+n\u03b3\u0398)exp((-E)/(1+\u03b3\u03b8))\u03a6.Reviewer #2: Recommendation: Minor revisionAuthors should revise the manuscript according to the following comments.\u2022 Nomenclature is must.Response: The nomenclature has been included in the revised manuscript.\u2022 Some grammatical and typo mistakes are found in the manuscript.Response: In the revised manuscript we have correct the grammatical issues.\u2022 The governing differential equations describing the flow are non-linear. Is the solution obtained unique?Response: Please if you focus on the graphs and boundary conditionsU_=0, V_=0,\u0398_ =1, \u03a6_=1 at Y_j=0 U_\u21920, \u3016 \u03b8\u3017_\u21920, \u03a6_(i.j)\u21920 as Y_j\u2192\u221e In these graphs and all other graphs given in this study, the solutions are satisfying by the boundary conditions, it is evident of the correctness of the obtained results. \u2022 Specify the applications of considered physical model or geometry.Response: The proposed model is used as the guidelines for the selection of a suitable coupling to achieve the desired applications. The curved shaped geometry is used to design many problems of civil engineering as pressure barrier.\u2022 Introduction section should be made more concise to show previous work in the field. At present a lot of related researches are stated in the introduction. However, not sufficient analysis is presented. The authors should ask themselves: what are the problems with the presented researches? Why is the recent work needed? Hope these can improve the present work by following past articles.\u2022 Effect of Joule heating on MHD non\u2010Newtonian fluid flow past an exponentially stretching curved surface\u2022 Magneto hydrodynamic Cattaneo-Christov flow past a cone and a wedge with variable heat source/sink\u2022 Heat and mass transfer in MHD Casson nanofluid flow past a stretching sheet with thermophoresis and Brownian motion.\u2022 Effect of asymmetrical heat rise/fall on the film flow of magneto hydrodynamic hybrid Ferro fluid\u2022 Simultaneous solutions for first order and second order slips on micro polar fluid flow across a convective surface in the presence of Lorentz force and variable heat source/sink.\u2022 Effect of thermal radiation on MHD Casson fluid flow over an exponentially stretching curved sheet\u2022 Influence of non-uniform heat source/sink on the three-dimensional magneto hydrodynamic Carreau fluid flow past a stretching surface with modified Fourier\u2019s law\u2022 MHD Carreau Fluid Flow Past a Melting Surface with Cattaneo-Christov Heat Flux\u2022 Physical aspects on unsteady MHD\u2010free convective stagnation point flow of micro polar fluid over a stretching surface\u2022 Influence of viscous dissipation on MHD flow of micro polar fluid over a slandering stretching surface with modified heat flux model\u2022 A non\u2010Fourier heat flux model for magneto hydrodynamics micro polar liquid flow across a coagulated sheetNote: I want to see the revised version of the manuscript.Response: Most relevant papers are added to enrich introduction (see in introduction ref [23] - [30]) in the revised manuscript.AttachmentResponse to Reviewers Comments.docxSubmitted filename: Click here for additional data file. 28 Apr 2021PONE-D-21-04681R1Effects of Temperature Dependent Viscosity and Thermal Conductivity on Natural Convection Flow along a Curved Surface in the Presence of Exothermic Catalytic Chemical ReactionPLOS ONEDear Dr. Ali,Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE\u2019s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.Please submit your revised manuscript by Jun 12 2021 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at Please include the following items when submitting your revised manuscript:A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.A marked-up copy of your manuscript that highlights changes made to the original version. 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Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see:\u00a0We look forward to receiving your revised manuscript.Kind regards,Naramgari Sandeep, Ph.DAcademic EditorPLOS ONEJournal Requirements:Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article\u2019s retracted status in the References list and also include a citation and full reference for the retraction notice.Reviewers' comments:Reviewer's Responses to Questions6. Review Comments to the AuthorPlease use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. Reviewer #1:\u00a0The report presents the natural convection boundary layer flow of a viscous incompressible fluid with temperature-dependent viscosity and temperature-dependent thermal conductivity in the presence of exothermic catalytic chemical reaction along a curved surface. I recommend minor revision due to the following facts.Q1. In Eq. (3), the authors added (u^2/2x)n to the convective acceleration term.Comment: What does the new term means? How did you derive it.In the report, the derivation presented by the Authors is in isolation. You are expected to derive it as a convective acceleration of momentum equation.Q2. After Eq. (6), it was written that n = P(x) + Q(x). Whereas is the wall temperature function andthe is the body shape function.This is the exact contribution of the report to the body of knowledge and it should be introduced under the introduction section. In fact, a paragraph is needed to announce this novelty.Q3. How do you study a fluid flow along a vertical surface without buoyancy forces? This is true because there is nothing like the associated dimensionless parameter called buoyancy parameter or Grashof number.Q4. Before Eq. 7, was defined as the tangential component of acceleration due to gravity. What is r in the definition?Reviewer #2:\u00a0The revisions are Good but some references are not arranged properly. Hence I recommend for possible publication.https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at\u00a0figures@plos.org. Please note that Supporting Information files do not need this step.While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool,\u00a0 14 May 2021Reply to Review PONE-D-21-04681R1Effects of Temperature Dependent Viscosity and Thermal Conductivity on Natural Convection Flow along a Curved Surface in the Presence of Exothermic Catalytic Chemical Reaction PLOS ONEReviewer #1: The report presents the natural convection boundary layer flow of a viscous incompressible fluid with temperature-dependent viscosity and temperature-dependent thermal conductivity in the presence of exothermic catalytic chemical reaction along a curved surface. I recommend minor revision due to the following facts.Q1. In Eq. (3), the authors added (u^2/2x)n to the convective acceleration term.Comment: What does the new term means? How did you derive it. In the report, the derivation presented by the Authors is in isolation. You are expected to derive it as a convective acceleration of momentum equation.Response: The detail derivation of momentum equation is given as below:The dimensioned form of momentum equation in article is given as underu \u2202u/\u2202x+v \u2202u/\u2202y=1/\u03c1 \u2202/\u2202y (\u03bc \u2202u/\u2202y)+g_x \u03b2_T (T-T_\u221e )+g_x \u03b2_C (C-C_\u221e) (1)Dimensionless variables x \u00ae=x/l,y \u00ae=y/l Gr^(1/4),u \u00ae=u/U_s ,v \u00ae=v/V_s Gr^(1/4),\u03b8=(T-T_\u221e)/(T_w-T_\u221e ) ,\u03d5 =(C-C_\u221e)/(C_w-C_\u221e ) (2)To convert the equation (1) into dimensionless form we use the dimensionless variables defined in equation (2), for this we find the term s one by one appeared in the equation (1)u \u2202u/\u2202x=U_s u \u00ae (\u2202U_s u \u00ae)/(\u2202x \u00ael)=(U_s^2)/l u \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+\u3016U_s u \u00ae\u3017^2/l (\u2202U_s)/(\u2202x \u00ae )u \u2202u/\u2202x=(U_s^2)/l u \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+\u3016U_s u \u00ae\u3017^2/l \u2202/(\u2202x \u00ae ) (g_x \u03b2_T \u0394T)^(1/2)u \u2202u/\u2202x=(U_s^2)/l u \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+\u3016U_s u \u00ae\u3017^2/l (g_x \u03b2_T \u0394T)^(1/2) 1/(2\u221a(g_x )) (dg_x)/(dx \u00ae ) u \u2202u/\u2202x=(U_s^2)/l [u \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+u \u00ae^2/2 1/g_x (dg_x)/(dx \u00ae )] (3)Nowv \u2202u/\u2202y=U_s v \u00aeGr^(-1/4) (\u2202U_s u \u00ae)/(\u2202ly \u00aeGr^(-1/4) )=(U_s^2)/l v \u00ae (\u2202u \u00ae)/(\u2202y \u00ae ) (4)Now1/\u03c1 \u2202/\u2202y (\u03bc \u2202u/\u2202y)=1/\u03c1 [\u2202\u03bc/\u2202y \u2202u/\u2202y+\u03bc (\u2202^2 u)/(\u2202y^2 )] (5)Considering \u2202\u03bc/\u2202y=\u2202/\u2202y \u03bc_o (1+\u03b3^* (T-T_\u221e))\u2202\u03bc/\u2202y=\u2202/(\u2202y \u00aelGr^(-1/4) ) (\u03bc_o (1+\u03b3^* \u0394T\u03b8))\u2202\u03bc/\u2202y=(\u03bc_o \u03b3^* \u0394T)/(lGr^(-1/4) ) \u2202\u03b8/(\u2202y \u00ae )\u2202\u03bc/\u2202y=(\u03bc_o \u03b3_T)/(lGr^(-1/4) ) \u2202\u03b8/(\u2202y \u00ae ) (6)\u2202u/\u2202y=(\u2202U_s u \u00ae)/(\u2202y \u00aelGr^(-1/4) )=U_s/(lGr^(-1/4) ) (\u2202u \u00ae)/(\u2202y \u00ae ) (7)(\u2202^2 u)/(\u2202y^2 )=(\u2202U_s u \u00ae)/(\u2202y \u00aelGr^(-1/4) )=U_s/(lGr^(-1/2) ) (\u2202^2 u \u00ae)/(\u2202y \u00ae^2 ) (8)Put eqs. (6)-(8) in eq. (5) we have 1/\u03c1 \u2202/\u2202y (\u03bc \u2202u/\u2202y)=1/\u03c1 [(\u03bc_o \u03b3_\u03bc)/(lGr^(-1/4) ) \u2202\u03b8/(\u2202y \u00ae ) U_s/(lGr^(-1/4) ) (\u2202u \u00ae)/(\u2202y \u00ae )+(\u03bcU_s)/(lGr^(-1/2) ) (\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )]1/\u03c1 \u2202/\u2202y (\u03bc \u2202u/\u2202y)=1/\u03c1 [(\u03bc_o \u03b3_\u03bc U_s)/(l^2 Gr^(-1/2) ) \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+\u3016\u03bc_o (1+\u03b3_\u03bc \u03b8)U\u3017_s/(l^2 Gr^(-1/2) ) (\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )]1/\u03c1 \u2202/\u2202y (\u03bc \u2202u/\u2202y)=(\u03bc_o U_s)/(\u03c1l^2 Gr^(-1/2) ) [\u03b3_\u03bc \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+(1+\u03b3_\u03bc \u03b8)(\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )]Where \u03b3_(\u03bc=) \u03b3^* \u0394T1/\u03c1 \u2202/\u2202y (\u03bc \u2202u/\u2202y)=(\u03bc_o U_s)/(\u03c1l^2 Gr^(-1/2) ) [\u03b3_\u03bc \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+(1+\u03b3_\u03bc \u03b8)(\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )] (9)Put the eqs. (3-4) and Eq. (9) in eq. (1)(U_s^2)/l [u \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+u \u00ae^2/2 1/g_x (dg_x)/(dx \u00ae )+v \u00ae (\u2202u \u00ae)/(\u2202y \u00ae )]=(\u03bc_o U_s Gr^(1/2))/(\u03c1l^2 ) [\u03b3_\u03bc \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+(1+\u03b3_\u03bc \u03b8)(\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )]+g_x \u03b2_T \u0394T\u03b8+g_x \u03b2_C \u0394C\u03d5 [u \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+u \u00ae^2/2 1/g_x (dg_x)/(dx \u00ae )+v \u00ae (\u2202u \u00ae)/(\u2202y \u00ae )]=(\u03bc_o Gr^(1/2))/(\u03c1lU_s ) [\u03b3_\u03bc \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+(1+\u03b3_\u03bc \u03b8)(\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )]+(\u3016lg\u3017_x \u03b2_T \u0394T\u03b8)/(U_s^2 )+(\u3016lg\u3017_x \u03b2_C \u0394T\u03d5)/(U_s^2 ) u \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+u \u00ae^2/2 1/g_x (dg_x)/(dx \u00ae )+v \u00ae (\u2202u \u00ae)/(\u2202y \u00ae )=(\u03bdGr^(1/2))/(lU_s ) [\u03b3_\u03bc \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+(1+\u03b3_\u03bc \u03b8)(\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )]+(\u3016lg\u3017_x \u03b2_T \u0394T\u03b8)/(U_s^2 )+(\u3016lg\u3017_x \u03b2_C \u0394C\u03d5)/(U_s^2 )u \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+u \u00ae^2/2 1/g_x (dg_x)/(dx \u00ae )+v \u00ae (\u2202u \u00ae)/(\u2202y \u00ae )=(\u03bdGr^(1/2))/(lU_s ) [\u03b3_\u03bc \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+(1+\u03b3_\u03bc \u03b8)(\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )]+(\u3016lg\u3017_x \u03b2_T \u0394T\u03b8)/(U_s^2 )+(\u3016lg\u3017_x \u03b2_C \u0394C\u03d5)/(U_s^2 )Where U_s=(g_x \u03b2_T \u0394Tl)^(1/2),Gr=(g_x \u03b2_T \u0394Tl^3)/\u03bd^2 and U_sc=(g_x \u03b2_T \u0394Cl)^(1/2),Gr^*=(g_x \u03b2_T \u0394Cl^3)/\u03bd^2 So u \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+u \u00ae^2/2 1/g_x (dg_x)/(dx \u00ae )+v \u00ae (\u2202u \u00ae)/(\u2202y \u00ae )=(\u03bdGr^(1/2))/(lU_s ) [\u03b3_\u03bc \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+(1+\u03b3_\u03bc \u03b8)(\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )]+(U_s^2 \u03b8)/(U_s^2 )+(U_s^2 \u03d5)/(U_s^2 )u \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+u \u00ae^2/2 1/g_x (dg_x)/(dx \u00ae )+v \u00ae (\u2202u \u00ae)/(\u2202y \u00ae )=\u03b3_\u03bc \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+(1+\u03b3_\u03bc \u03b8)(\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )+\u03b8+\u03d5 (10)Where the detail derivation of n=P(x)+Q(x) is given in below steps:Since we know thatd/dx (lnx)=1/xandd(lnx)=1/x dxwhich implies that1/(d(lnx))=x/dx. Also considerd/dx (ln g_x )=1/g_x (dg_x)/dxwhich impliesd(ln g_x )=1/g_x dg_x. (d lng_x)/(d lnx)=x/g_x (dg_x)/dx.Now using the non-dimensionless variables, we may write the above equations as(d lng_x)/(d lnx \u00ael)=(x \u00ael)/g_x (dg_x)/(dx \u00ael)which implies that(d lng_x)/(d lnx \u00ael)=x \u00ae/g_x (dg_x)/(dx \u00ae )and thus we have 1/x \u00ae (d lng_x)/(d lnx \u00ael)=1/g_x (dg_x)/(dx \u00ae ) Also, we consider d/dx (lnT_w )=1/T_w (dT_w)/dx=0which implies that d lnT_w=0and thus, we can write (d lnT_w)/(d lnx \u00ael)=0 Now from equation (c) and (d), we have1/x \u00ae ((d lnT_w)/(d lnx \u00ae l)+(d lng_x)/(d lnx \u00ae l))=1/g_x (dg_x)/(dx \u00ae )which can also be written as 1/x \u00ae (P(x \u00ae)+Q(x \u00ae))=1/g_x (dg_x)/(dx \u00ae ) Here, P(x \u00ae), the wall temperature function and Q(x \u00ae) the body shape function, are defined asP(x \u00ae )=(d lnT_w)/(d lnx \u00ae l), Q(x \u00ae )=(d lng_x)/(d lnx \u00ae l) . Now (10) becomesu \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+u \u00ae^2/2(P(x \u00ae )+Q(x \u00ae))+v \u00ae (\u2202u \u00ae)/(\u2202y \u00ae )=\u03b3_\u03bc \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+(1+\u03b3_\u03bc \u03b8)(\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )+\u03b8+\u03d5 (11) Now P(x \u00ae )+Q(x \u00ae )=nThus (11) becomesu \u00ae (\u2202u \u00ae)/(\u2202x \u00ae )+u \u00ae^2/2 1/g_x (dg_x)/(dx \u00ae )+v \u00ae (\u2202u \u00ae)/(\u2202y \u00ae )=\u03b3_\u03bc \u2202\u03b8/(\u2202y \u00ae ) (\u2202u \u00ae)/(\u2202y \u00ae )+(1+\u03b3_\u03bc \u03b8)(\u2202^2 u \u00ae)/(\u2202y \u00ae^2 )+\u03b8+\u03d5 Which is dimensionless form of momentum equation along curved surface, and n is the body shape parameter.Q2. After Eq. (6), it was written that n = P(x) + Q(x). Whereas is the wall temperature function andthe is the body shape function. This is the exact contribution of the report to the body of knowledge and it should be introduced under the introduction section. In fact, a paragraph is needed to announce this novelty.Response: In current paper we are investigated the natural convection flow over a two dimensional body of arbitrary geometric configuration in the presence of exothermic catalytic chemical reaction. The momentum, energy, and mass concentration equations are a general form suitable for laminar natural convection flows along curved surface in the inclusion of exothermic catalytic reaction. For body of arbitrary shape, the special case in which P(x \u00ae )+Q(x \u00ae )=n, thus the body shape parameter (index parameter) n has chosen 0\u20090.05). In addition, the HK\u03b1\u03b1/\u03b1\u03b1 was mainly distributed in Qiandongnan, but its distribution had no statistically significant difference between those regions (p\u2009>\u20090.05). The \u03b1\u03b1\u03b1anti3.7/\u2013 SEA was quite rare in those regions, and only one case was detected in Qiandongnan.As listed in Table anti3.7/\u2013SEA case, and 18 cases of the HK\u03b1\u03b1/\u03b1\u03b1 did not presented any clinical manifestations such as anemia at the time of examination. The blood parameters including the red blood cells (RBC), hemoglobin (HGB), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), HbA, HbA2, and HbF were measured and statistically analyzed through ANOVA. No significant parameter difference was identified among the three different genotype groups (p\u2009>\u20090.05) (Tables 0 (HBB: c.126_129delCTTT) and \u03b1\u03b1/\u03b1\u03b1\u03b1anti4.2 (Table 0 (HBB:c.52A\u2009>\u2009T) and the \u03b1\u03b1/\u03b1\u03b1\u03b1anti4.2.The frequency of thalassemia-gene carriers in Guizhou is 11.03%, with 7.41% of \u03b1-thalassemia-gene frequency and 3.23% of \u03b2-thalassemia-gene frequency (unpublished data). Therefore, the frequency of \u03b1-thalassemia is higher than \u03b2-thalassemia in Guizhou province, China. While deletion of \u03b1-globin genes causes \u03b1-thalassemia, the triplicated \u03b1-globin genes alone rarely cause obvious clinical symptoms. All 59 carriers of the triplicated \u03b1-globin genes including the \u03b1\u03b1\u03b1anti4.2/\u03b1\u03b1, \u03b1\u03b1\u03b1anti3.7/\u03b1\u03b1. Although the HK\u03b1\u03b1 shows by GAP-PCR the positivity for \u03b1\u03b1\u03b1anti4.2 and -\u03b13.7, it is not considered as \u03b1-globin triplication due to no real extra copy of the \u03b1-globin gene existed in the HK\u03b1\u03b1 allele. Observations have indicated that \u03b1-globin triplication alone does not cause detectable clinical manifestations [anti3.7 and \u03b1\u03b1\u03b1anti4.2 was different between the two subpopulations. In our study, the ratio of \u03b1\u03b1\u03b1anti3.7/\u03b1\u03b1\u03b1anti4.2 was 0.47 (0.248% \u03b1\u03b1\u03b1anti3.7/0.523% \u03b1\u03b1\u03b1anti4.2) in Guizhou province, while it was 3.0 (0.9% \u03b1\u03b1\u03b1anti3.7/0.3% \u03b1\u03b1\u03b1anti4.2) in Guangdong province. This finding suggests that the \u03b1\u03b1\u03b1anti4.2 triplication is rather common in Guizhou, while the \u03b1\u03b1\u03b1anti3.7 is prevalent in Guangdong region.To date, at least two genotypes of \u03b1-globin triplication have been described: \u03b1\u03b1\u03b1stations . Howeverstations , and in anti4.2 in Tujia was significantly higher (1.65%) than in any other ethnic group. This is the first report that Tujia have a higher frequency of \u03b1-globin triplication. Whether this higher rate of \u03b1-globin triplication is caused by a founder effect or not requires further investigation. Although previous studies have reported that the frequencies of \u03b1-globin defects in the Zhuang and Yao ethnic groups were significantly higher than that in the Han ethnic group in Guangxi, another province in southwestern China [Ethnically, the Tujia group presented the highest prevalence (2.47%) of the \u03b1-globin triplication. In particular, the prevalence of \u03b1\u03b1\u03b1rn China , we did anti4.2 in Qiannan was also higher than in other regions. The other type of \u03b1-globin triplication, \u03b1\u03b1\u03b1anti3.7/\u2013SEA, and the non-\u03b1-triplication HK\u03b1\u03b1/\u03b1\u03b1 had no significant geographical differences. In addition, we did not identify any anti-HK\u03b1\u03b1 allele, although we found that the frequency of HK\u03b1\u03b1 allele was 0.235%, significantly higher than previously reported. To exclude the presence of HK\u03b1\u03b1 in case of positivity for the -\u03b13.7 and \u03b1\u03b1\u03b1anti4.2, next-generation and Gap-PCR are needed to be simultaneously performed. Interestingly, while the HK\u03b1\u03b1 is hardly observed in other regions, it was relatively common in Qiandongnan and its frequency is comparable to that of the \u03b1\u03b1\u03b1anti 4.2 triplication there . The frequency difference of the \u03b1-globin triplication between Qiannan and any other region was statistically significant. Moreover, the frequency of \u03b1\u03b1\u03b1%. The franti4.2, and \u03b1\u03b1\u03b1anti3.7 carriers were all within the normal range, which is consistent with previous reports [anti4.2, and \u03b1\u03b1\u03b1anti3.7 will not present clinical manifestations such as anemia. Of note, although the HK\u03b1\u03b1 carriers presented a normal range of hematological parameters, their RBC, hemoglobin, MCH, and MCV were all slightly lower than the \u03b1\u03b1\u03b1anti4.2 and \u03b1\u03b1\u03b1anti3.7 carriers, implying that the particular cluster structure that could reduce the \u03b1-globin gene expression.In our study, the hematological parameters and hemoglobin electrophoresis data of the HK\u03b1\u03b1, \u03b1\u03b1\u03b1 reports , 22. Theanti3.7/\u2013SEA did not presented any clinical manifestations such as anemia at the time of examination, which is consistent with previous reports [As mentioned above, the triplicated \u03b1-globin genes alone barely lead to detectable clinical phenotypes. In this study, of the 59 cases of \u03b1-globin genes triplication, 5 cases coinherited with \u03b2-globin gene mutation(s) while the other 54 subjects did not present any clinical symptoms. Although many reports have stated that \u03b1-globin triplication can exacerbate the symptoms of \u03b2-thalassemia, the issue is still controversial because the expected worsened anemia has not occurred in all cases , 18. In reports , 23.Currently, Gap-PCR and PCR combined with RDB (reverse dot blot) methods are commonly used to detect \u03b1-globin gene deletions and the \u03b2-globin gene defects, but they usually miss the triplicated \u03b1-globin genes. In this study, whole genome NGS combined with Gap-PCR was adopted to screen for all types of \u03b1-globin and \u03b2-globin gene alterations, including \u03b1-globin gene deletion, triplication, splicing mutations, which would be expected to increase the detection sensitivity and improve the diagnosis of \u03b2-thalassemia.This epidemiological study has identified the current \u03b1-triplication genotypes and their prevalence and distribution in Guizhou province, which will be used to update the triplicated \u03b1-globin thalassemia database, provide insights into the pathogenesis of thalassemia and shed light on the diagnosis of thalassemia in southwestern China."} +{"text": "The title compound crystallizes with half of a mol\u00adecule per asymmetric unit and exhibits bond lengths and angles typical of \u03b1-diketones. A network of C\u2014H\u22efF contacts and \u03c0\u2013\u03c0 stacking inter\u00adactions is observed within the structure. 14H6F4O2, crystallizes with half of a mol\u00adecule per asymmetric unit and exhibits bond lengths and angles typical of \u03b1-diketones. A network of C\u2014H\u22efF contacts and \u03c0\u2013\u03c0 stacking inter\u00adactions is observed within the structure.The title compound, C The title \u03b1-diketone, 1,2-bis\u00adethane-1,2-dione, is used as a precursor in the production of hexa\u00adbenzocoronenes and similar \u03b1,\u03b1-diketones crystallize in trigonal or monoclinic space groups, respectively , and exhibits the expected bond lengths and angles for \u03b1-diketone sp2 hybridized atoms. Inter\u00adestingly, the C5\u2014C6\u2014C7\u2014O1 torsion angle [7.55\u2005(19)\u00b0] of the title compound is larger compared to the same torsion angle in bis\u00ad(4-fluoro\u00adphen\u00adyl)ethane-1,2-dione . A network of C\u2014H\u22efO inter\u00adactions is also observed between the carbonyl oxygen and H5. This inter\u00adaction is significantly weaker for 1,2-bis\u00adethane-1,2-dione in comparison to benzil (O\u22efH = 2.42\u2005\u00c5) and bis\u00ad(4-fluoro\u00adphen\u00adyl)ethane-1,2-dione (O\u22efH = 2.40\u2005\u00c5). As a result, the \u03c0\u2013\u03c0 stacking and C\u2014H\u22ef F inter\u00adactions play a vital role in how the compound packs within the crystal structure.A view of crystal packing of the title compound is presented in Fig.\u00a02ne Fig.\u00a03. Similaret al., 2016A search of the Cambridge Structural Database pentane-1,4-dione.Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021005363/yk2148sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021005363/yk2148Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021005363/yk2148Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021005363/yk2148Isup4.cmlSupporting information file. DOI: 2085161CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular conformation is stabilized by an intra\u00admolecular O\u2014H\u22efO hydrogen bond between the hy\u00addroxy substituent on the benzene ring and one of the carbonyl groups of the acridinedione unit. 27H33NO5, a 3,3,6,6-tetra\u00admethyl\u00adtetra\u00adhydro\u00adacridine-1,8-dione ring system carries an ethyl acetate substituent on the acridine N atom and an o-hy\u00addroxy\u00adphenyl ring on the central methine C atom of the di\u00adhydro\u00adpyridine ring. The benzene ring is inclined to the acridine ring system at an angle of 80.45\u2005(7)\u00b0 and this conformation is stabilized by an intra\u00admolecular O\u2014H\u22efO hydrogen bond between the hy\u00addroxy substituent on the benzene ring and one of the carbonyl groups of the acridinedione unit. The ester C=O oxygen atom is disordered over major and minor orientations in a 0.777\u2005(9):0.223\u2005(9) ratio and the terminal \u2013CH3 unit of the ethyl side chain is disordered over two sets of sites in a 0.725\u2005(5): 0.275\u2005(5) ratio. In the crystal, C\u2014H\u22efO hydrogen bonds combine to link the mol\u00adecules into a three-dimensional network. van der Waals H\u22efH contacts contribute the most to the Hirshfeld surface (66.9%) followed by O\u22efH/H\u22efO (22.1%) contacts associated with weak hydrogen bonds.In the title compound, C In contrast, the central C13/N1/C1/C6\u2013C8 ring can best be described as a flattened boat with N1 and C7 displaced by 0.146\u2005(1) and 0.191\u2005(14)\u2005\u00c5, respectively, from the remaining four C atoms. The bond lengths and angles in the title mol\u00adecule agree reasonably well with those found in closely related mol\u00adecules . The relative contributions of the other inter\u00adactions in descending order are: O\u22efH/H\u22efO (22.1%), C\u22efH/H\u22efC (9.2%), O\u22efO (1.3%), N\u22efH/H\u22efN (0.2%) and N\u22efC/C\u22efN (0.2%). This illustrates that the C\u2014H\u22efO inter\u00adactions contribute significantly to the crystal packing.The overall two-dimensional fingerprint plot is illustrated in Fig.\u00a04ts Fig.\u00a04b. The rH)-yl]acetic acid [Cam\u00adbridge Structural Database -yl]acetate -dione -dione -1,8-dioxo-2,3,4,5,6,7,8,9-octa\u00adhydro\u00adacridin-10(1S(8) ring. In the crystal, the mol\u00adecules are linked by O\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional network. In VANBUK, the central 1,4-di\u00adhydro\u00adpyridine ring adopts a shallow sofa conformation (with the C atom bearing the bromo\u00adphenol ring as the flap), whereas the pendant cyclo\u00adhexene rings both have twisted-boat conformations. The mol\u00adecule features an intra\u00admolecular O\u2014H\u22efO hydrogen bond, which closes an S(8) ring. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO inter\u00adactions, forming C(12) chains propagating along the c-axis direction. In the crystal of SILBIB, O\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22ef\u03c0(ring) hydrogen bonds combine with an Br\u2014O and unusual C\u2014Br\u22ef\u03c0(ring) halogen bonds to generate a three dimensional network with mol\u00adecules stacked along the a-axis direction. In the acridinedione moiety of PUSJEU, the central di\u00adhydro\u00adpyridine ring adopts a flattened-boat conformation, with the N atom and the methine C atom displaced from the mean plane of the other four atoms by 0.0513\u2005(14) and 0.1828\u2005(18)\u2005\u00c5, respectively. The two cyclo\u00adhexenone rings adopt envelope conformations, with the tetra\u00adsubsituted C atoms as the flap atoms. In the crystal, mol\u00adecules are linked via a pair of C\u2014H\u22efO hydrogen bonds, forming inversion dimers, which are, in turn, linked by C\u2014H\u22efO hydrogen bonds, forming slabs lying parallel to (001).The DABSAD compound crystallizes with two mol\u00adecules in the asymmetric unit. In each mol\u00adecule, the central 1,4-di\u00adhydro\u00adpyridine ring adopts a shallow sofa conformation (with the C atom bearing the phenol ring as the flap), whereas the pendant cyclo\u00adhexene rings both have twisted-boat conform\u00adations. Each mol\u00adecule features an intra\u00admolecular O\u2014H\u22efO hydrogen bond, which closes an To a mixture of dimedone , ethyl glycinate hydro\u00adchloride and salicaldehyde in ethanol (20\u2005ml), triethyl amine was added. The reaction mixture was heated under reflux for 5\u2005h at 353\u2013358\u2005K then left to cool. The separated solid was filtered off, dried and recrystallized from ethanol solution as yellow plates of the title compound, yield 68%, m.p. 497\u2005K.Uiso(H) = 1.2Ueq (C) and O\u2014H = 0.84\u2005\u00c5, Uiso(H) = 1.5Ueq (O). Atom O3 of the oxo group and terminal methyl group (C17) of the ethyl acetate substituent are disordered over two sites in 0.777\u2005(9):0.223\u2005(9) (for O3 and O3A) and 0.725\u2005(5):0.275\u2005(5) (for C17 and C17A) ratios, respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021001341/hb7967sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989021001341/hb7967Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021001341/hb7967Isup3.cmlSupporting information file. DOI: 2061379CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A linear supra\u00admolecular chain featuring amine-N\u2014H\u22efO(nitro) hydrogen bonding is noted in the crystal.The N 22H17N4O2S)2], comprises two N,S-donor anions, leading to a distorted tetra\u00adhedral N2S2 donor set. The resultant five-membered chelate rings are nearly planar and form a dihedral angle of 73.28\u2005(3)\u00b0. The configurations about the endocyclic- and exocyclic-imine bonds are Z and E, respectively, and that about the ethyl\u00adene bond is E. The major differences in the conformations of the ligands are seen in the dihedral angles between the chelate ring and nitro\u00adbenzene rings [40.48\u2005(6) cf. 13.18\u2005(4)\u00b0] and the N-bound phenyl and nitro\u00adbenzene ring [43.23\u2005(8) and 22.64\u2005(4)\u00b0]. In the crystal, a linear supra\u00admolecular chain along the b-axis direction features amine-N\u2014H\u22efO(nitro) hydrogen bonding. The chains assemble along the 21-screw axis through a combination of phenyl-C\u2014H\u22efO(nitro) and \u03c0(chelate ring)\u2013\u03c0(phen\u00adyl) contacts. The double chains are linked into a three-dimensional architecture through phenyl-C\u2014H\u22efO(nitro) and nitro-O\u22ef\u03c0(phen\u00adyl) inter\u00adactions.The title zinc bis\u00ad(thio\u00adsemicarbazone) complex, [Zn(C These ligands usually act as monodentate or bidentate ligands and coordinate with transition and non-transition metal ions either in neutral or anionic form through thione/thiol\u00adate-sulfur and azomethine/imine-nitro\u00adgen donor atoms \u00b0 for the S1\u2014Zn\u2014N1 chelate angle, to a wide 131.16\u2005(2)\u00b0, for S1\u2014Zn\u2014S2, consistent with an approximate tetra\u00adhedral geometry. The mode of coordination of the thio\u00adsemicarbazone ligands leads to the formation of five-membered chelate rings. These are nearly planar with r.m.s. deviations of 0.0459 and 0.0152\u2005\u00c5 for the S1- and S2-containing rings, respectively. However, the maximum deviation from the plane through the S1-chelate ring of \u22120.0613\u2005(9)\u2005\u00c5 for the N1 atom suggests an alternate description of the conformation of the S1-ring might be valid. Another description might be an envelope conformation with the zinc atom lying 0.209\u2005(3)\u2005\u00c5 out of the plane of the four remaining atoms (r.m.s. deviation = 0.0005\u2005\u00c5). The dihedral angle between the mean plane through the rings is 73.28\u2005(3)\u00b0. There are three formal double bonds in each thio\u00adsemicarbazone anion. Owing to chelation, the configuration about the endocyclic imine bond is Z whereas that about the exocylic imine bond is E; the configuration of the ethyl\u00adene bond is E.The mol\u00adecular structure of (I)N-bound phenyl and nitro\u00adbenzene rings is 72.41\u2005(5), 16.96\u2005(11) and 40.48\u2005(6)\u00b0, respectively, for the S1-ring compared with 82.47\u2005(6), 20.33\u2005(5) and 13.18\u2005(4)\u00b0, respectively, for the S2-ring. Similarly, the pairs of dihedral angles between the imine- and N-bound phenyl rings, i.e. 59.15\u2005(6) and 76.48\u2005(8)\u00b0, and N-bound phenyl and nitro\u00adbenzene rings, i.e. 43.23\u2005(8) and 22.64\u2005(4)\u00b0, show notable differences; the dihedral angles between the imine-phenyl and nitro\u00adbenzene rings are comparable, i.e. 82.28\u2005(7) and 85.67\u2005(7)\u00b0. Finally, the nitro groups present different relative orientations with respect to the benzene rings they are connected to, with the N4-nitro group being twisted out of the plane. This is shown in the value of the C2\u2014C3\u2014N4\u2014O1 torsion angle of 161.88\u2005(18)\u00b0 compared with \u22120.4\u2005(3)\u00b0 for the C26\u2014C25\u2014N8\u2014O3 torsion angle.Some major differences are noted in the conformations of the ligands. Thus, the sequence of dihedral angles formed between the chelate ring and the imine-phenyl, b-axis direction, Table\u00a02a). The hydrogen bonds involve the N7-amine, there being no apparent role for the N3-amine in the supra\u00admolecular aggregation. A phenyl-C44\u2014H\u22efO4(nitro) contact provides extra stability to the chain and indicates the nitro-O4 atom forms two contacts. Chains assemble about the 21-screw axis via a combination of phenyl-C37\u2014H\u22efO3(nitro) and \u03c0\u2013\u03c0 contacts. The \u03c0\u2013\u03c0 contacts are of particular inter\u00adest in that the participating rings are a phenyl and a chelate ring, as highlighted in Fig.\u00a02b); such inter\u00adactions are now well recognized in the supra\u00admolecular chemistry of metal complexes and impart significant energies of stabilization to the packing \u22efCgi is 3.5559\u2005(11)\u2005\u00c5 with an inter-planar angle = 6.70\u2005(8)\u00b0 and slippage of 0.34\u2005\u00c5 for symmetry operation (i): x, y, z. The links between chains to consolidate the three-dimensional architecture are of the type phenyl-C14\u2014H\u22efO1(nitro) and nitro-O1\u22ef\u03c0(phen\u00adyl), Table\u00a02Cg(C23\u2013C28)ii = 3.4788\u2005(19)\u2005\u00c5 with angle at O1 = 108.71\u2005(13)\u00b0 for (ii): 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z. A view of the unit-cell contents is shown in Fig.\u00a03Conventional amine-N7\u2014H\u22efO4(nitro) hydrogen bonds are noted in the crystal of (I)Crystal Explorer 17 dnorm-mapped Hirshfeld surface of Fig.\u00a06dnorm-mapped Hirshfeld surface. However, the \u03c0\u2013\u03c0 inter\u00adaction appears as a flat surface on the curvedness-mapped Hirshfeld surface of Fig.\u00a08a), the nitro-O\u22ef\u03c0 inter\u00adaction is shown as red concave and blue bump regions on the shape-index-mapped Hirshfeld surface of Fig.\u00a08b).The faint-red spots observed on the a)\u2013(f), respectively. The percentage contributions from each inter\u00adatomic contact are summarized in Table\u00a04b), with the peak tipped at de = di \u223c2.2\u2005\u00c5 corresponding to the H24\u22efH44 contact, Table\u00a03de + di \u223c2.0 and 2.6\u2005\u00c5, respectively, Fig.\u00a09c) and (d). The H\u22efS/S\u22efH contacts contribute 8.6% and appear as two blunt-symmetric wings at de + di \u223c2.9\u2005\u00c5 in Fig.\u00a09e). This feature reflects the long-range H\u22efS/S\u22efH contact evinced in the packing with a separation of 0.1\u2005\u00c5 shorter than the sum of their van der Waals radii, Table\u00a03de + di \u223c2.6\u2005\u00c5 in the fingerprint plot of Fig.\u00a09f), the contribution to the overall Hirshfeld surface is only 5.2%. The other 11 inter\u00adatomic contacts have a negligible effect on the mol\u00adecular packing as their accumulated contribution is below 11%, Table\u00a04The overall two-dimensional fingerprint plot for (I)d,p) level of theory. The total energy (Etot) was calculated by summing four energy components, comprising the electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies. The independent energy components as well as the Etot are tabulated in Table\u00a05Edis energy term still makes the major contribution to the inter\u00adaction energies partly due to the presence of \u03c0\u2013\u03c0, N\u2014O\u22ef\u03c0, C\u2014H\u22efO and C\u2014H\u22efC inter\u00adactions. The total Edis components of all pairwise inter\u00adactions sum to \u2212432.1\u2005kJ\u2005mol\u22121, whereas the total Eele sums to \u2212190.2\u2005kJ\u2005mol\u22121. The stabilization of the crystal through the contribution of the dispersion forces is emphasized by the energy framework diagram, Fig.\u00a010b axis.The pairwise inter\u00adaction energies between mol\u00adecules in the mol\u00adecular packing of (I)N-bound ethyl species with a terminal phenyl ring -N-phenyl\u00adhydrazine-1-carbo\u00adthio\u00adamide, (VI), was filtered, washed with cold ethanol and dried in vacuo after which it was used without further purification. Compound (VI) was dissolved in hot absolute ethanol (50\u2005ml), which was added to a solution of Zn(CH3COO)2\u00b72H2O in hot absolute ethanol (40\u2005ml). The mixture was heated (348\u2005K) and stirred for about 10\u2005min, followed by stirring for about 1\u2005h at room temperature. The white precipitate obtained was filtered, washed with cold ethanol and dried in vacuo. Single crystals were grown at room temperature by slow evaporation of (I)v/v 20\u2005ml). Yield: 90%, m.p. 511\u2013512\u2005K. FT\u2013IR (ATR (solid) cm\u22121): 3428 \u03bd(N\u2014H), 1593 \u03bd(C=N), 1335 \u03bd(N\u2014N), 579 \u03bd(Zn\u2014N), 489 \u03bd(Zn\u2014S). UV\u2013Visible: \u03bbmax ): 250 , 292 , 433 . ICP\u2013AES: Experimental %Zn = 7.26, Theoretical %Zn = 7.53.Analytical grade reagents were used as procured and without further purification. 4-Phenyl-3-thio\u00adsemicarbazide and 4-nitro\u00adchalcone were dissolved separately in hot absolute ethanol (50\u2005ml) and mixed while stirring. About five drops of concentrated hydro\u00adchloric acid were added to the mixture and the mixture was heated (348\u2005K) while stirring for about 30\u2005min. The yellow precipitate, (2Uiso(H) set to 1.2Ueq(C). The N-bound H atoms were located in a difference-Fourier map, but were refined with an N\u2014H = 0.88\u00b10.01\u2005\u00c5 distance restraint, and with Uiso(H) set to 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S2056989021007398/mw2178sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989021007398/mw2178Isup2.hklStructure factors: contains datablock(s) I. DOI: 2097106CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dihedral angle between the aromatic rings in the title compound is 31.84\u2005(8)\u00b0; N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions connect mol\u00adecules in the crystal, producing a three-dimensional network. 16H12F5N3O, the dihedral angle between the aromatic rings is 31.84\u2005(8)\u00b0. In the crystal, the mol\u00adecules are linked into dimers possessing crystallographic twofold symmetry by pairwise N\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22efO hydrogen bonds and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions link the dimers into a three-dimensional network. A Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from F\u22efH/H\u22efF (41.1%), H\u22efH (21.8%), C\u22efH/H\u22efC (9.7%) C\u22efC (7.1%) and O\u22efH/H\u22efO (7.1%) contacts. The contribution of some disordered solvent to the scattering was removed using the SQUEEZE routine in PLATON. The solvent contribution was not included in the reported mol\u00adecular weight and density.In the title compound, CSpek 2015. Acta Cr R)C=N\u2014NHR grouping possess controllable E/Z isomerization around the C=N double bond, which makes them good candidates for the construction of functional materials \u2005\u00c5. The backbone of the mol\u00adecule is non-planar with a dihedral angle of 31.84\u2005(8)\u00b0 between the C1\u2013C6 penta\u00adflouro\u00adbenzene and C11\u2013C16 benzene rings and the acetamide group lies almost perpendicular. The C5\u2014C6\u2014C7\u2014N2, C6\u2014C7\u2014N2\u2014N3, C7\u2014N2\u2014N3\u2014C11, N2\u2014N3\u2014C11\u2014C16 and C6\u2014C7\u2014C8\u2014N1 torsion angles are \u221228.19\u2005(17), 174.02\u2005(10), \u2212176.33\u2005(11), 5.90\u2005(18) and 122.80\u2005(12)\u00b0, respectively.The title mol\u00adecule Fig.\u00a01 crystallCg1\u22efCg1b = 3.7137\u2005(10)\u2005\u00c5, slippage = 1.158\u2005\u00c5, Cg1\u22efCg2b = 3.7015\u2005(9)\u2005\u00c5, slippage = 1.407\u2005\u00c5, and Cg1\u22efCg2a = 3.7016\u2005(9)\u2005\u00c5, slippage = 1.148\u2005\u00c5; where Cg1 and Cg2 are the centroids of the C1\u2013C6 and C11\u2013C16 rings, respectively; symmetry codes: (a) 1\u00a0\u2212\u00a0x, y, z; (b) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z]. Together, these generate a three-dimensional network in blue and negative electrostatic potential is indicated in red (hydrogen-bond acceptors).a. The percentage contributions to the Hirshfeld surfaces from various inter\u00adatomic contacts (Table\u00a02b), H\u22efH , C\u22efH/H\u22efC C\u22efC and O\u22efH/H\u22efO . Other contact types including N\u22efH/H\u22efN, N\u22efC/C\u22efN and N\u22efN contacts account for less than 5.4% of the Hirshfeld surface mapping and presumably have minimal directional impact on the packing.The overall two-dimensional fingerprint map for the title compound is shown in Fig.\u00a04s Table\u00a02 are F\u22efH/s Table\u00a02b, H\u22efH -1-benzyl\u00adidene-2-phenyl\u00adhydrazine skeleton are (E)-3-chloro-N\u2032-(2-fluoro\u00adbenzyl\u00adidene)thio\u00adphene-2-carbohydrazide ethyl\u00adidene]isonicotinohydrazide -bis\u00ad[(thio\u00adphen-2-yl)meth\u00adyl\u00adidene]hydrazine ethyl\u00adidene]nicotinohydrazide \u2005\u00c5. The mol\u00adecular skeleton is approximately planar, the terminal five- and six-membered rings forming a dihedral angle of 5.47\u2005(9)\u00b0. In the crystal, mol\u00adecules are linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds into zigzag chains propagating in [100].The hydrazide derivative SOJQAL adopts an E conformation with respect to the azomethine bond. The pyridyl and fluoro\u00adbenzene rings make dihedral angles of 38.58\u2005(6) and 41.61\u2005(5)\u00b0 respectively with the central C(=O)N2CC unit, resulting in a non-planar mol\u00adecule. The inter\u00admolecular inter\u00adactions comprise two classical N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds and four non-classical C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds. These inter\u00adactions are augmented by a weak \u03c0\u2013\u03c0 inter\u00adaction between the benzene and pyridyl rings of neighbouring mol\u00adecules, with a centroid\u2013centroid distance of 3.9226\u2005(10)\u2005\u00c5. This leads to a three-dimensional supra\u00admolecular assembly in the crystal.The mol\u00adecule of HIXRAJ adopts an The asymmetric unit of MIHROK03 comprises two independent half-mol\u00adecules, each residing on a centre of symmetry. The two mol\u00adecules are essentially planar. In the crystal, weak C\u2014H\u22ef\u03c0 inter\u00adactions join the two symmetry-independent mol\u00adecules into inter\u00adlinked chains parallel to [011].E conformation with respect to the azomethine double bond whereas the N and methyl C atoms are in a Z conformation with respect to the same bond. The ketonic O and azomethine N atoms are cis to each other. The non-planar mol\u00adecule [the dihedral angle between the benzene rings is 7.44\u2005(11)\u00b0] exists in an amido form with a C=O bond length of 1.221\u2005(2)\u2005\u00c5. In the crystal, a bifurcated N\u2014H\u22ef hydrogen bond is formed between the amide H atom and the keto O and imine N atoms of an adjacent mol\u00adecule, leading to the formation of chains propagating along the b-axis direction.The mol\u00adecule of ZISSAX adopts an via hydrogen bonding of the carboxyl groups. In addition, there is an N\u2014H\u22efO hydrogen bond between the amino group and the carbonyl O atom.In TINWIX, the aromatic rings are almost perpendicular, making a dihedral angle of 89.26\u2005(5)\u00b0. The carboxyl group is coplanar with the aromatic ring to which it is attached [dihedral angle = 1.70\u2005(17)\u00b0]. The packing involves inversion-symmetric dimers bridged E)-1-[(perfluoro\u00adphen\u00adyl)methyl\u00adene]-2-phenyl\u00adhydrazine , tetra\u00admethyl\u00adethylenedi\u00adamine (TMEDA) , CuCl and CCl4 . After 1\u20133\u2005h , the reaction mixture was poured into a 0.01 M solution of HCl , and extracted with di\u00adchloro\u00admethane (3 \u00d7 20\u2005ml). The combined organic phase was washed with water (3 \u00d7 50\u2005ml), brine (30\u2005ml), dried over anhydrous Na2SO4 and concentrated in vacuo using a rotary evaporator. The residue was purified by column chromatography on silica gel using appropriate mixtures of hexane and di\u00adchloro\u00admethane (3/1\u20131/1). Colourless prisms of the title compound suitable for X-ray analysis were obtained by slow evaporation of a di\u00adchloro\u00admethane solution (69%); m.p. 405\u2005K. Analysis calculated for C16H12F5N3O: C 53.79, H 3.39, N 11.76; found: C 53.73, H 3.36, N 11.71%. 1H NMR \u03b4 3.04 , 6.50\u20137.33 . 13C NMR \u03b4 33.58, 108.97, 116.87, 120.75, 124.11, 124.76, 140.95, 146.33, 149.87, 150.91, 155.21. ESI\u2013MS: m/z: 358.24 [M + H]+.A 20\u2005ml screw-neck vial was charged with DMSO (10\u2005ml), (Uiso(H) = 1.2 or 1.5Ueq (C). The residual electron density was difficult to model and therefore the SQUEEZE routine contains approximately three electrons.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021007349/hb7979sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989021007349/hb7979Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021007349/hb7979sup3.docxElectrostatic potential map. DOI: Click here for additional data file.10.1107/S2056989021007349/hb7979Isup4.cmlSupporting information file. DOI: 1878189CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III complex was synthesized from the substitution reaction between the (ppy)2Ir(\u03bc-Cl)2Ir(ppy)2 dimer and 1,1-bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)methane under an argon gas atmosphere. The IrIII atom is coordinated by two C,N-bidentate ppy anions, a unidentate dppm ligand and a chloride anion in a distorted octa\u00adhedral IrC2N2PCl arrangement.The title Ir III complex, [Ir(C11H8N)2Cl(C25H22P2)], was synthesized from the substitution reaction between the (ppy)2Ir(\u03bc-Cl)2Ir(ppy)2 dimer and 1,1-bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)methane under an argon gas atmosphere for 20\u2005h. The IrIII atom is coordinated by two C,N-bidentate ppy anions, a unidentate dppm ligand and a chloride anion in a distorted octa\u00adhedral IrC2N2PCl arrangement. The N donor atoms of the ppy ligands are mutually trans while the C atoms are cis. Intra\u00admolecular aromatic \u03c0\u2013\u03c0 stacking between the phenyl rings of ppy and dppm, and C\u2014H\u22efCl inter\u00adactions are observed. In the crystal, C\u2014H\u22efCl and C\u2014H\u22ef\u03c0 contacts link the mol\u00adecules into a three-dimensional network. A Hirshfeld surface analysis was carried out to further qu\u00adantify the inter\u00admolecular inter\u00adactions, and indicated that H\u22efH contacts (63.9%) dominate the packing.The title Ir The bond lengths of Ir1\u2014N1, Ir1\u2014N2, Ir1\u2014C11 and Ir1\u2014C22 are 2.051\u2005(2), 2.062\u2005(2), 2.004\u2005(3) and 2.032\u2005(3)\u2005\u00c5, respectively. As expected, the averaged Ir\u2013C and Ir\u2014N bond lengths are much shorter than the Ir\u2014Cl and Ir\u2014P bonds, based on the sizes of the different species.The asymmetric unit of (I)2(dppm)]PF6 e.g., Hao et al., 20192(dppm)](PF6) iridium(III) are reported to be 2.503\u2005(19) trans bond angles deviate from the ideal 180\u00b0 [170.97\u2005(9)\u2013175.37\u2005(8)\u00b0], similar to related compounds methane (dppm) often occurs as a bidentate ligand (Cg5\u22efCg7 = 3.621\u2005(1)\u2005\u00c5 and between the phenyl rings of the dppm mol\u00adecule (C29\u2013C34 and C36\u2013C41), Cg8\u22efCg9 = 3.997\u2005(1)\u2005\u00c5 \u2005\u00c5], C30\u2014H30\u22efCl1 [C\u22efCl = 3.664\u2005(4)\u2005\u00c5] and C35\u2014H35A\u22efCl1 [C35\u22efCl = 3.460\u2005(3)\u2005\u00c5] \u2005\u00c5 Fig.\u00a02. Three w\u00c5] Fig.\u00a03 are obse\u22ef\u03c0 (ring) inter\u00adactions are found in the crystal packing of the C17\u2013C22 phenyl ring (H3\u22efCg6 = 2.83\u2005\u00c5). In addition, C\u2014H\u22ef\u03c0(ring) inter\u00adactions are also found between the dppm phenyl rings of neighbouring mol\u00adecules: C26\u2014H26 \u22efCg10 , C38\u2014H38\u22efCg10 (H38\u22efCg10 = 2.81\u2005\u00c5) and C40\u2014H40\u22efCg8 (H40\u22efCg8 = 2.95\u2005\u00c5). In addition, pairwise inter\u00admolecular hydrogen bonds are observed between C14\u2014H14 of the pyridine ring of the ppy ring and Cl1 with the functions de and di, which are the distances from an indicated area on the Hirshfeld surface to the nearest atoms outside and inside the surface, respectively. The white, red, and blue areas on the dnorm-mapped Hirshfeld surfaces show inter\u00admolecular contacts that are equal to, shorter than, and longer than the sum of their van der Waals (vdW) radii, respectively. A pair of inter\u00admolecular contacts are shown as red spots on the Hirshfeld surface close to the Cl1 atom of the adjoining mol\u00adecule and the H14 atom of the associated pyridine ring. The spots indicate hydrogen-bond donor-to-acceptor inter\u00adactions of C14\u2014H14\u22efCl1 and vice versa while Fig.\u00a06b)\u2013(d) depict the contacts of the H\u22efH (63.9%), C\u22efH/H\u22efC (29.5%) and H\u22efCl/Cl\u22efH (4.4%) inter\u00adactions, respectively.Additional insights into the weak inter\u00admolecular contacts in the crystal packing of (I)sa Fig.\u00a05. The relSciFinder ]+; dppm = bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)methane bidentate ligand. However, none of the remaining publications describe a monomeric IrIII complex similar to the title compound. The seven hits include the octa\u00adhedral crystal structures of IrIII complexes with a bis\u00ad(2-phenyl\u00adpyridine)\u00adiridium(III) backbone and ancillary ligands of both N-donor, P-donor and O-donor ligands. There are a tris-complex of Ir(ppy)3 ]; dppel = 1,2-bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)ethyl\u00adene, [Ir(ppy)2(dppp)]; dppp = 1,3-bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)propane and [Ir(ppy)2(dppe)]; dppe = 1,2-bis\u00ad(diphenylphosphan\u00adyl) ethane] ]PF6, [Ir(dfppy)2(P^N)]PF6 and [Ir(dfmppy)2(P^N)]PF6 where P^N = 2-[(di\u00adphenyl\u00adphos\u00adphan\u00adyl) meth\u00adyl]pyridine, dfppy = 2-pyri\u00addine and dfmppy = 2--4-methyl\u00adpyridine ]PF6 and refined as riding with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021000955/hb7961sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021000955/hb7961Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021000955/hb7961Isup3.molSupporting information file. DOI: 2058995CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, generating 22H15N3O4, is built up from a central imidazopyridine ring system connected to a nitroso group, a phenyl ring and a 2-oxo-2-phenyl\u00adethyl acetate group. The imidazo pyridine ring system is almost planar (r.m.s. deviation = 0.017\u2005\u00c5) and forms dihedral angles of 22.74\u2005(5) and 45.37\u2005(5)\u00b0, respectively, with the phenyl ring and the 2-oxo-2-phenyl\u00adethyl acetate group. In the crystal, the mol\u00adecules are linked into chains parallel to the b axis by C\u2014H\u22efO hydrogen bonds, generating R21 (5) and R44 (28) graph-set motifs. The chains are further linked into a three-dimensional network by C\u2014H\u22ef\u03c0 and \u03c0-stacking inter\u00adactions. The inter\u00admolecular inter\u00adactions were investigated using Hirshfeld surface analysis and two-dimensional fingerprint plots, revealing that the most important contributions for the crystal packing are from H\u22efH (36.2%), H\u22efC/C\u22efH (20.5%), H\u22efO/O\u22efH (20.0%), C\u22efO/O\u22efC (6.5%), C\u22efN/N\u22efC (6.2%), H\u22efN/N\u22efH (4.5%) and C\u22efC (4.3%) inter\u00adactions.The title compound, C N-heterocycles as the core structure, including imidazopyridine and its derivatives, which are used in medicinal chemistry Numerous drugs contain a] pyridine ring system is planar with an r.m.s deviation of 0.017\u2005\u00c5 and a maximum deviation of 0.028\u2005(1)\u2005\u00c5 for atom C11. The mean plane through the fused ring system makes dihedral angles of 22.74\u2005(5) and 45.37\u2005(5)\u00b0 with the phenyl ring (C1\u2013C6) and the 2-oxo-2-phenyl\u00adethyl acetate group (C14\u2013C22), respectively. The dihedral angle between the two aromatic rings (C1\u2013C6 and C17\u2013C22) is 59.63\u2005(5)\u00b0. The mol\u00adecular conformation is stabilized by two weak intra\u00admolecular C9\u2014H9\u22efO1 and C1\u2014H1\u22efN1 hydrogen bonds, generating S(6) ring motifs (Table\u00a01The mol\u00adecular structure of (I)s Table\u00a01.iii and C10\u2014H10\u22efO2iii hydrogen bonds, forming chains that propagate parallel to the b axis and enclose A\u22efO4i and C15\u2014H15B\u22efO1ii hydrogen bonds with Cg4iv inter\u00adaction (Cg4 is the centroid of the C17\u2013C22 phenyl ring) as well as \u03c0\u2013\u03c0 stacking inter\u00adactions involving the centroids (Cg1 and Cg2) of the N2/C13/N3/C7\u2013C8 and N2/C9\u2013C13 rings with a centroid-to-centroid distance Cg1\u22efCg2 of 3.5750\u2005(9)\u2005\u00c5 and a slippage of 0.685\u2005\u00c5 N-acetamide carbonoimido\u00adyl]phenol \u00b0. In TUQCEP, C21H17N3O, the fused ring system is almost planar (r.m.s. deviation = 0.031\u2005\u00c5) and forms dihedral angles of 64.97\u2005(7) and 18.52\u2005(6)\u00b0 with the phenyl ring and the (imino\u00admeth\u00adyl)phenol group, respectively. In its crystal, mol\u00adecules are linked by pairs of C\u2014H\u22ef\u03c0 inter\u00adactions into centrosymmetric dimeric units, which are further connected by O\u2014H\u22efN hydrogen bonds, forming layers parallel to (101).A search of the Cambridge Structural Database to 1.2371 (blue) a.u. . The shape-index map of the title mol\u00adecule was generated in the range \u22121 to 1\u2005\u00c5 , revealing the presence of red and blue triangles that are indicative of the presence of \u03c0\u2013\u03c0 stacking inter\u00adactions. The curvedness map of the title complex was generated in the range \u22124.0 to 4.0\u2005\u00c5 and shows flat surface patches characteristic of planar stacking. The Hirshfeld surface representations with the function dnorm plotted onto the surface are shown for the H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH, C\u22efO/O\u22efC, C\u22efN/N\u22efC, H\u22efN/N\u22efH and C\u22efC inter\u00adactions in Fig.\u00a04a\u2013g, respectively. The overall two-dimensional fingerprint plot is illustrated in Fig.\u00a05a, with those delineated into H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH, C\u22efO/O\u22efC, C\u22efN/N\u22efC, H\u22efN/N\u22efH and C\u22efC contacts associated with their relative contributions to the Hirshfeld surface in Fig.\u00a05b\u2013h, respectively. The most important inter\u00admolecular inter\u00adaction is H\u22efH, contributing 36.2% to the overall crystal packing . H\u22efC/C\u22efH contacts, with a 20.5% contribution to the Hirshfeld surface, indicate the presence of the weak C\u2014H\u22ef\u03c0 inter\u00adaction . H\u22efO/O\u22efH contacts arising from inter\u00admolecular C\u2014H\u22efO hydrogen bonding make a 20.0% contribution to the Hirshfeld surface and are represented by a pair of sharp spikes in the region de + di \u223c2.34\u2005\u00c5 . The C\u22efC contacts are a measure of \u03c0\u2013\u03c0 stacking inter\u00adactions and contribute 4.3% of the Hirshfeld surface . The contributions of the other contacts to the Hirshfeld surface are C\u22efO/O\u22efC of 6.5%, C\u22efN/N\u22efC of 6.2% and H\u22efN/N\u22efH of 4.5%.Hirshfeld surface analysis was used to qu\u00adantify the inter\u00admolecular contacts of the title compound, using u. Fig.\u00a03a. The s\u2005\u00c5 Fig.\u00a03b, revea\u2005\u00c5 Fig.\u00a03c and shng Fig.\u00a05b. H\u22efC/Cn Table\u00a01. Two paint Fig.\u00a05c. H\u22efO/O\u2005\u00c5 Fig.\u00a05d. The Cce Fig.\u00a05h. The ca]pyridine-8-carboxyl\u00adate in acetic acid (50\u2005ml), sodium nitrite was added at room temperature. The resulting precipitate was washed with water and extracted with di\u00adchloro\u00admethane (3 \u00d7 20\u2005ml). The combined di\u00adchloro\u00admethane extracts were dried over anhydrous sodium sulfate and filtered. The remaining solution was concentrated under reduced pressure. The residue was purified chromatographically on a neutral alumina gel column using di\u00adchloro\u00admethane as eluent. Single crystals were obtained by slow evaporation of a di\u00adchloro\u00admethane solution at room temperature (yield 80%).To a solution of 2-oxo-2-phenyl\u00adethyl 2-phenyl\u00adimidazo, C\u2014H = 0.93\u2005\u00c5 for aromatic [Uiso(H) = 1.2Ueq(C)] and C\u2014H = 0.98\u2005\u00c5 for methine [Uiso(H) = 1.2Ueq(C)] H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022001517/wm5632sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022001517/wm5632Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022001517/wm5632Isup3.cmlSupporting information file. DOI: 2106558CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(6) ring motif.The title compound is a Schiff base that exists in the phenol\u2013imine tautomeric form. The mol\u00adecular structure is stabilized by an O\u2014H\u22efN hydrogen bond, forming an 24H24N2O4, was synthesized by the inter\u00adaction of 2-hy\u00addroxy-3-meth\u00adoxy benzaldehyde and 1,4-benzene dimethanamine in ethanol, and crystallizes in the monoclinic space group P21/n with Z\u2032 = 0.5. The mol\u00adecule is not planar, the 1,4-di\u00adethyl\u00adbenzene and the phenol rings are twisted with respect to each other, making a dihedral angle of 74.27\u2005(5)\u00b0. The mol\u00adecular structure is stabilized by an O\u2014H\u22efN hydrogen bond, forming an S(6) ring motif. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, resulting in the formation of sheets parallel to the bc plane. A Hirshfeld surface analysis was undertaken to investigate the various inter\u00admolecular contacts controlling the supra\u00admolecular topology, suggesting the H\u22efO (18%) contacts to be the most significant inter\u00adactions, whereas the H\u22efH (50.5%) and C\u22efH (24.3%) inter\u00adactions are less significant.The Schiff base compound, C There is an intra\u00admolecular O2\u2014H2\u22efN1 hydrogen bond (Table\u00a01S(6) ring motif and also induces the phenol ring and the Schiff base to be nearly coplanar, as indicated by the C6\u2014C8\u2014N1\u2014C9 torsion angle of 178.54\u2005(13)\u00b0. The mol\u00adecule is non-planar, the 1,4-di\u00adethyl\u00adbenzene ring being inclined to the phenol ring by 74.27\u2005(5)\u00b0. The C7\u2014C6\u2014C8\u2014N1 torsion angle [3.8\u2005(2)\u00b0] further supports the co-planarity of the phenol ring and the Schiff base. The C7\u2014O2 distance is 1.3438\u2005(17)\u2005\u00c5, which is close to normal values reported for single C\u2014O bonds in phenols and salicyl\u00adidene\u00adamines moiety revealed some related structures. The most similar structures are 1,4-bis\u00ad(2-pyridyl\u00admethyl\u00adene\u00adamino\u00admeth\u00adyl)benzene benzene ring motif as in the title compound. The length of intra\u00admolecular O\u2014H\u22efN hydrogen bond in OCAPAK is especially short [1.65\u2005(2)\u2005\u00c5] compared to that in the title compound [1.789\u2005(15)\u2005\u00c5].A search of the Cambridge Structural Database using CrystalExplorer17 to 1.404 a.u. (blue). The packing of mol\u00adecules is mainly dependent on H\u22efH (50.5%) and C\u22efH (24.3%) inter\u00adactions and the significant C\u2014H\u22efO inter\u00adactions (18%). Blue regions in the dnorm map indicate inter\u00admolecular inter\u00adactions with distances longer than van der Waals radius sum of the inter\u00adacting elements of 2-hy\u00addroxy-3-meth\u00adoxy benzaldehyde was dissolved in 20\u2005mL of ethanol and mixed with 0.0100\u2005g (0.074\u2005mmol) of 1,4-benzene dimethanamine dissolved in 20\u2005mL of ethanol Fig.\u00a04. The reaUiso(H) = 1.5Ueq(O) and a distance restraint. The C-bound H atoms were positioned geometrically and refined using a riding model, with Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021013347/jq2011sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021013347/jq2011Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021013347/jq2011Isup3.cmlSupporting information file. DOI: 2128953CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the organic dication and perchlorate anions are linked through N\u2014H\u22efO, C\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, forming a three-dimensional network.In the title salt, C 20H42N42+\u00b72ClO4\u2212, has been determined using synchrotron radiation at 220\u2005(2)\u2005K. The structure determination reveals that protonation has occurred at diagonally opposite amine N atoms. The asymmetric unit comprises one half of the organic dication, which lies about a center of inversion, and one perchlorate anion. The macrocyclic dication adopts the most stable endodentate trans-III conformation. The crystal structure is stabilized by intra\u00admolecular N\u2014H\u22efN, and inter\u00admolecular N\u2014H\u22efO and C\u2013H\u22efO hydrogen bonds involving the macrocycle N\u2014H and C\u2014H groups as donors and the O atoms of perchlorate anions as acceptors, giving rise to a three-dimensional network.The crystal structure of the title salt, C The macrocycle is basic and readily captures two or four protons to form the [C20H42N4]2+ dication or the [C20H44N4]4+ tetra\u00adcation in which all of the N\u2014H bonds are generally available for hydrogen-bond formation ](NO3)2\u00b73H2O, [Cu(C20H40N4)](NO3)2, [Cu(C20H40N4)](ClO4)2 and [Cu(C20H40N4)(H2O)2](BF4)2\u00b72H2O were reported together with [Zn(C20H40N4)(OCOCH3)2]. In these structures, the copper(II) or zinc(II) cations have tetra\u00adgonally distorted octa\u00adhedral environments with the four N atoms of the macrocyclic ligand in equatorial positions and the O atoms of the counter-anions, water mol\u00adecules or acetato ligands in axial positions \u00b72(C11H10O) \u00b72(NO2OH) \u00b72MeOH 2, (I)We report here the preparation of a new dicationic compound, [Canti with respect to the macrocyclic plane as a result of the mol\u00adecular inversion symmetry. The [C20H42N4]2+ dication adopts an endodentate conformation and trans-III configuration along the center of the macrocyclic cavity. The endo conformation of the dication may be due to the intra\u00admolecular N\u2014H\u22efN hydrogen-bonding inter\u00adaction. Within the centrosymmetric diprotonated amine unit, the C\u2014C and N\u2014C bond lengths range from 1.5173\u2005(18) to 1.5368\u2005(18)\u2005\u00c5 and from 1.4795\u2005(16) to 1.5044\u2005(16)\u2005\u00c5, respectively. The range of N\u2014C\u2014C and C\u2014N\u2014C angles is 108.89\u2005(11) to 113.50\u2005(11)\u00b0 and 113.46\u2005(11) to 114.61\u2005(11)\u00b0, respectively. The bond lengths and angles within the dication are comparable to those found in the free ligand or other cations in (C20H40N4)\u00b72C11H10O \u00b72MeOH 3}(CN)3]2\u00b72H2O\u00b72MeOH \u2005\u00c5] and N1\u2014C3 [1.4795\u2005(16)\u2005\u00c5] are slightly shorter than N2\u2014C8 [1.5044\u2005(16)\u2005\u00c5] and N2\u2014C9 [1.4952\u2005(18)\u2005\u00c5]. Each of the two hydrogen atoms of N2 and N2\u2032 is involved in hydrogen bonding with both of the two remaining nitro\u00adgen atoms (Table\u00a014\u2212 anion vary from 1.4218\u2005(19) to 1.4529\u2005(16)\u2005\u00c5, and the O\u2014Cl\u2014O angles vary from 106.45\u2005(10) to 110.51\u2005(12)\u00b0. The distorted geometry of the ClO4\u2212 anion undoubtedly results from its involvement in hydrogen-bonding inter\u00adactions with the organic cation.An ellipsoid plot of the mol\u00adecular components in (I)s Table\u00a01. The int4\u2212 anions are connected to the [C20H42N4]2+ dication by N\u2014H\u22efO hydrogen bonds. The macrocyclic dication is linked to a neighboring ClO4\u2212 anion through a very weak C\u2014H\u22efO hydrogen bond. The extensive array of these contacts generates a three-dimensional network structure , [C20H42N4]2+ or [C20H44N4]4+. The crystal structures of (C20H40N4)\u00b72C11H10O \u00b72MeOH were used as provided. All chemicals were reagent grade and used without further purification. As a starting material, macrocycle 3,14-dimethyl-2,6,13,17-tetra\u00adaza\u00adtri\u00adcyclo\u00addocosane, L, was prepared according to a published procedure was suspended in methanol (20\u2005mL) and the pH was adjusted to 3.0 with 0.5 M HClO4. The mixture was stirred magnetically for 30\u2005min and the resulting solution was filtered. The neat filtrate was allowed to stand for one week to give block-like colorless crystals of (I)Commercially available A and H2B) were placed in geometrically idealized positions and constrained to ride on their parent atoms, with C\u2014H distances of 0.97\u20130.98\u2005\u00c5 and an N\u2014H distance of 0.99\u2005\u00c5, and with Uiso(H) values of 1.5 and 1.2 times, respectively, that of the parent atoms. The one N-bound H atom (H1N1) of the amine was assigned based on a difference-Fourier map, and a Uiso(H) value of 1.5Ueq(N1).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021004278/vm2247sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021004278/vm2247Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021004278/vm2247Isup3.cmlSupporting information file. DOI: 2079010CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the mol\u00adecules are linked by C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds into strips, which are connected by F\u22efF contacts into layers. a]pyridine core in the mol\u00adecule of the title compound, C10H7F3N2O, is planar within 0.004\u2005(1)\u2005\u00c5. In the crystal, the mol\u00adecules are linked by pairs of C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds, forming strips. These strips are connected by the F\u22efF contacts into layers, which are further joined by \u03c0\u2013\u03c0 stacking inter\u00adactions. The Hirshfeld surface analysis and fingerprint plots reveal that mol\u00adecular packing is governed by F\u22efH/H\u22efF (31.6%), H\u22efH (16.8%), C\u22efH/H\u22efC (13.8%) and O\u22efH/H\u22efO (8.5%) contacts.The bicyclic imidazo[1,2- The C2\u2014C1\u2014C8\u2014C9 and N2\u2014C1\u2014C8\u2014O1 torsion angles of 1.04\u2005(18) and 1.14\u2005(19)\u00b0, respectively, show that the ethanone group lies near the plane of the bicycle. The bond lengths N1\u2014C2, C2\u2014C1 and C1\u2014C8 of 1.3367\u2005(16), 1.3987\u2005(16) and 1.4247\u2005(16)\u2005\u00c5, respectively, indicate strong \u03c0-conjugation in the N1\u2013O1 chain.In the mol\u00adecule of the title compound Fig.\u00a01, the fusCg1\u22efCg1; Cg1 is the centroid of the imidazole ring].In the crystal, the mol\u00adecules are linked by pairs of C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds into strips elongated along the [210] direction Figs. 2. These ss Figs. 3. The lays Figs. 3 and \u03c0\u2013\u03c0 Crystal Explorer 17 imidazopyridin-3-amine imidazopyridin-3-amine ring motif. In the crystal, a short H\u22efH contact links adjacent mol\u00adecules into centrosymmetric dimers. The dimers are joined by weak C\u2014H\u22ef\u03c0 and slipped \u03c0\u2013\u03c0 stacking inter\u00adactions, forming layers parallel to (110), which are connected into a three-dimensional network by short Br\u22efH contacts. In the crystal of PILGAV01, N\u2014H\u22efN hydrogen bonds link the mol\u00adecules into [010] chains. The cohesion of the crystal structure of DABTEI is ensured by C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds, forming layers parallel to the ac plane. In ZAPJAD, the supra\u00admolecular structure is defined by two kinds of inter\u00admolecular hydrogen bonds. Pairs of N\u2014H\u22efN hydrogen bonds link the mol\u00adecules into centrosymmetric dimers and N\u2014H\u22efO hydrogen bonds link the dimers into tubular chains running along the a-axis direction. In the crystal of ULEGOI, mol\u00adecules are linked into chains through pairs of C\u2014H\u22efN inter\u00adactions, forming a axis.The most closely related compounds containing a similar imidazo+.A mixture of (Uiso(H) = 1.2Ueq(C) for CH hydrogen atoms and Uiso(H) = 1.5Ueq(C) for CH3 hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021012676/yk2161sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021012676/yk2161Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021012676/yk2161Isup3.cmlSupporting information file. DOI: 2124974CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E)-2-benzyl-1,3-di\u00adphenyl\u00adiso\u00adthio\u00aduronium iodide, was prepared by the reaction of 1,3-di\u00adphenyl\u00adthio\u00adurea and benzyl iodide. In the crystal, N\u2014H\u22efI hydrogen bonds link the components into [100] chains.The title compound, a salt form of \u00b0 and electrons are delocalized over the N+= C\u2014N skeleton. The dihedral angle between the aromatic rings attached to the N atoms is 40.60\u2005(9)\u00b0. In the crystal, N\u2014H\u22efI hydrogen bonds link the components into [100] chains.In the title mol\u00adecular salt, C R\u2013S\u2013C\u2013(NHR)2+ moiety have been investigated as their hydrogen\u2013bonding motifs for mol\u00adecular recognition of anions \u00admethyl\u00adene]benzenaminium and toluyl units are linked to the sulfur atom as a thio\u00adether. The C7\u2014S1 and C8\u2014S1 bond lengths are 1.823\u2005(2) and 1.751\u2005(2)\u2005\u00c5, respectively, and the C\u2014S\u2014C bond angle is 101.66\u2005(9)\u00b0. The conformation of C1 and C8 about the C7\u2014S1 bond is gauche [C1\u2014C7\u2014S1\u2014C8 = 49.53\u2005(16)\u00b0]. The C\u2014S\u2014C bond angle in the title compound is somewhat smaller than that for di-p-tolyl sulfide \u00admethyl\u00adene]benz\u00aden\u00adaminium moiety of the title cation, the \u03c0-electrons of the iminium double bond are delocalized over the N1\u2014C6\u2014N2 skeleton .The title compound, CI\u2212 Fig.\u00a01, is a moIn the crystal, the cations and anions are linked by almost linear N\u2014H\u22efI hydrogen bonds Fig.\u00a02, generatvia CCDC Access Structures, November 2021; Groom et al., 2016N-[(phenyl\u00adamino)\u00admethyl\u00adene]benzenaminium but the compound most similar to the title compound is S-benzyl\u00adiso\u00adthio\u00aduronium chloride , to give a the title compound as a yellow solid . A solution of iso\u00adthio\u00aduronium iodide in methanol was slowly evap\u00adorated at room temperature to give crystals of the title compound: m.p. 442\u2013443\u2005K; 1H NMR : \u03b4 7.21\u20137.39 , \u03b4 4.45 ; HR TOF\u2013MS for C20H18N2S: calculated 318.1186 (M+), found 318.1185 (M+).1,3-Di\u00adphenyl\u00adthio\u00adurea (4.4\u2005mmol) was added to a solution of benzyl iodide (13.2\u2005mmol) in dry di\u00adchloro\u00admethane at room temperature. The reaction mixture was then stirred for 24\u2005h and concentrated Uiso(H) = 1.2Ueq(carrier).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021013086/hb7997sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021013086/hb7997Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021013086/hb7997Isup3.cmlSupporting information file. DOI: 2127354CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The parallel-aligned adtp4\u2212 ligands with an alternately reversed arrangement further link adjacent CoII ribbons into (010) layers, which are assembled into a three-dimensional supra\u00admolecular architecture via inter\u00admolecular hydrogen bonds.The title compound, {[Co 2(C12H7NO8)(H2O)2]\u00b71.6H2O}n comprises two CoII ions, which are coordinated by fully deprotonated 2-aminodi\u00adacetic terephthalic acid (adtp4\u2013) and terminal water mol\u00adecules in distorted octa\u00adhedral N1O5 and O6 coordination environments. The title compound features tetra\u00adnuclear CoII units bridged by \u03ba3O:O:O\u2032- and \u03ba3O:O,O\u2032-carboxyl\u00adate groups, which are joined into ribbons via syn\u2013anti carboxyl\u00adate bridges. The parallel adtp4\u2013 ligands with an alternately reversed arrangement further link adjacent CoII ribbons into (010) layers, which are assembled into a three-dimensional supra\u00admolecular network via inter\u00admolecular hydrogen bonds. The disordered water solvent mol\u00adecules are situated in channels parallel to [100]. Magnetic measurements and analyses reveal that the title compound displays anti\u00adferromagnetic behaviour. The purity of the title compound was characterized by X-ray powder diffraction.The asymmetric unit of the polymeric title compound {[Co The crystal structure, power X-ray diffraction pattern and magnetism of (I) were also studied in detail.Our previous studies have focused on the construction of CPs based on semi-rigid multi\u00adcarb\u00adoxy\u00adlic acids with the aminodi\u00adacetate moiety such as 2-aminodi\u00adacetic terephthalic acid (HI) comprises two CoII ions, one adtp4\u2013 ligand, two terminal water ligands and 1.6 disordered solvent water mol\u00adecules. Regarding the adtp4\u2013 ligand, one carboxyl\u00adate group of the aminodi\u00adacetate moiety adopts a \u03ba3-O:O:O\u2032 coordination mode and the other one employs a syn\u2013anti bidentate bridging fashion, whereas the carboxyl\u00adate group in the ortho-position coordinates in a \u03ba3-O:O,O\u2032 mode and that in the meta-position binds to one CoII ion in monodentate fashion (see Scheme). As shown in Fig.\u00a011O5 coordination set for Co1 and an O6 set for Co2. The adip4\u2013 ligand chelates Co1 with the amino nitro\u00adgen atom (N1) and carboxyl\u00adate oxygen atoms from the aminodi\u00adacetate moiety and its ortho-positioned carbox\u00adylate group. The residual cis-related sites are occupied by one meta-positioned carboxyl\u00adate oxygen atom (O4ii) and one aminodi\u00adacetate oxygen atom (O7i) from two other adip4\u2013 ligands (for symmetry codes refer to Table\u00a01ortho-positioned carboxyl\u00adate group (O1iii and O2iii) from another adip4\u2013 ligand chelates Co2, two cis-related positions of which are occupied by two aminodi\u00adacetate oxygen atoms (O8iv and O6) from two different adip4\u2013 ligands. The remaining two cis-related sites of Co2 are occupied by two terminal water ligands (O9 and O10). The length of the Co\u2014N bond is 2.241\u2005(3)\u2005\u00c5 and the Co\u2014O distances are between 1.992\u2005(3) and 2.362\u2005(3)\u2005\u00c5, which are all in the expected ranges. As shown in Fig.\u00a024\u2013 ligands bridge two pairs of CoII ions into a tetra\u00adnuclear unit with their \u03ba3O:O:O\u2032-carboxyl\u00adate groups from the aminodi\u00adacetate moieties and ortho-positioned \u03ba3O:O,O\u2032-carboxyl\u00adate groups from \u03ba3O:O:O\u2032-carboxyl\u00adate groups doubly bridge Co1 and Co1ii into a dinuclear unit. The dinuclear unit is further joined with two equivalent Co2i and Co2iii atoms via \u03ba3O:O:O\u2032-carboxyl\u00adate groups and \u03bc2-oxygen bridges (O1 and O1i) from \u03ba3O:O,O\u2032-carboxyl\u00adate groups. Adjacent tetra\u00adnuclear units are linked into a ribbon via double syn\u2013anti bridging carboxyl\u00adate groups from the aminodi\u00adacetate moieties. The closest Co1\u22efCo2 and Co1\u22efCo1 distances in the ribbon are 3.7074\u2005(8) and 3.5762\u2005(8)\u2005\u00c5, respectively. Parallel-aligned adtp4\u2013 ligands with an alternately reversed arrangement bind adjacent CoII ribbons into a layer extending parallel to (010) Fig.\u00a03.I) are assembled into a three-dimensional supra\u00admolecular network via inter\u00admolecular hydrogen bonds O9\u2014H9A\u22efO3v and O9\u2014H9B\u22efO2vi (Table\u00a02The (010) layers of (i Table\u00a02. The pos\u03c7M) of (I) were measured in the range 2\u2013300\u2005K under 1000 Oe. The \u03c7M, \u03c7M\u22121 and \u03c7MT versus T plots are shown in Fig.\u00a05\u03c7MT at 300\u2005K is 5.43\u2005cm3\u2005K\u2005mol\u22121, which is much larger than the expected spin-only value (3.75\u2005cm3\u2005K\u2005mol\u22121) of two isolated CoII ions with g = 2.0, S = 3/2, which may be due to the contribution of the incompletely quenched orbital magnetic moment. As the temperature decreases, the \u03c7MT value decreases slowly between 300 and 50\u2005K and then it descends more steeply to the minimum value of 0.51\u2005cm3\u2005K\u2005mol\u22121 at 2\u2005K. The curve clearly indicates that the dominant anti\u00adferromagnetic coupling is operating. The temperature dependence of \u03c7M\u22121 follows the Curie\u2013Weiss law, and the linear fit by the equation 1/\u03c7M = (T\u00a0\u2212\u00a0\u03b8)/C gives C = 5.76\u2005cm\u22123 K mol\u22121 and \u03b8 = \u221221.99\u2005K, which is consistent with an anti\u00adferromagnetic behaviour.The variable-temperature magnetic susceptibilities . The other two CoII complexes are discrete coordination mol\u00adecules (H2O)6]\u00b75H2O}n, has been synthesized, which consists of parallel stacked zigzag chains in which CoII cations are linked together through \u03bc3-adtp4\u2212 anions.A search of the Cambridge Structural Database of Co(NO3)2\u00b76H2O and 5.0\u2005ml of CH3CN, then transferred into a 25.0\u2005ml Teflon-lined stainless steel autoclave. The autoclave was sealed, heated to 393\u2005K and held at that temperature for 72\u2005h. The autoclave was allowed to cool to 303\u2005K within 24\u2005h. Plate-like pink crystals of (I) were collected in 66% yield based on H4adtp. Analysis calculated (%) for C12Co2N1O11.6H14.2 (Mr = 475.90): C 30.29, H 3.01, N, 2.94; found: C 30.18, H 3.15, N 3.06. Selected IR data : 3389 (s), 1631 (s), 1570 (m), 1405 (s), 1373 (s), 1319 (b), 1111 (b), 780 (b), 712 (b).HI) was confirmed by powder X-ray diffraction analysis . The peak positions of the experimental PXRD patterns are in good agreement with those simulated on basis of the present single-crystal X-ray data, indicating that a pure phase was obtained.The phase purity of compound (Uiso(H) = 1.5 Ueq(O). Other hydrogen atoms were placed at geometrically calculated positions and treated as riding, with Csp2\u2014H = 0.93\u2005\u00c5, Csp3\u2014H = 0.97\u2005\u00c5 and Uiso(H) = 1.2 Ueq(C). H atoms of O11, O12, O13 and O14 are not included in the model but were taken into account in the overall formula.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021008355/wm5615sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021008355/wm5615Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021008355/wm5615sup4.tifFigure S1 The simulated and experimental PXRD patterns for compound (I). DOI: 2063394CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of a ciprofloxacin salt with 2,6-di\u00adhydroxy\u00adbenzoic acid and a ciprofloxacin hydro\u00adchloride salt co-crystal with 3,5-di\u00adhydroxy\u00adbenzoic acid are reported. 17H19FN3O3+\u00b7C7H5O4\u2212, (I), and ciprofloxacin hydro\u00adchloride\u20133,5-di\u00adhydroxy\u00adbenzoic\u2013water (1/1/1), C17H19FN3O3+\u00b7Cl\u2212\u00b7C7H6O4\u00b7H2O, (II), were determined. In (I) and (II), the ciprofloxacin cations are connected via head-to-tail N\u2014H\u22efO hydrogen bonding. Both structures show an alternating layered arrangement between ciprofloxacin and di\u00adhydroxy\u00adbenzoic acid.The crystal structure of two multi-component crystals of ciprofloxacin [systematic name: 1-cyclo\u00adpropyl-6-fluoro-4-oxo-7-(piperazin-1-yl)quinoline-3-carb\u00ad\u00adoxy\u00adlic acid], a fluoro\u00adquinolone anti\u00adbiotic, namely, ciprofloxacin 2,6-di\u00adhydroxy\u00adbenzoate salt, C The formation of multi-component crystals, Ka of 1.30 and 1.325\u2005(3)\u2005\u00c5 indicate that it exists as the neutral carb\u00adoxy\u00adlic form. However, in 2,6HBA, the C\u2013O distances are very similar i.e., 1.263\u2005(4) and 1.267\u2005(3)\u2005\u00c5 due to resonance stabilization in the carboxyl\u00adate anion on Fig.\u00a01. The C\u2014OP1 space group despite the lack of a chiral centre. The asymmetric unit comprises one ciprofloxacin cation, one chloride anion and one 3,5HBA mol\u00adecule, as shown in Fig.\u00a02Ka of 4.04 and 1.314\u2005(4)\u2005\u00c5, respectively, also confirm the neutral state of this moiety. On the other hand, the piperazinyl group is protonated. Hence, compound (II)Compound (II)A\u22efF1 hydrogen bonding (Tables 1Compounds (I) Tables 1 and 2 \u25b8.i.e. N3\u2014H3B\u22efO4, N3\u2014H3B\u22efO5, and N3\u2014H3A\u22efO6, form an infinite chain structure along the a-axis direction a-axis, centrosymmetric pairs of ciprofloxacin mol\u00adecules are stacked by \u03c0\u2013\u03c0 inter\u00adactions. The distance between the centroids of symmetry-related C4\u2013C9 rings is 3.4986\u2005(11)\u2005\u00c5. This arrangement leads to the formation of a columnar packing arrangement. Inter\u00adestingly, a similar packing feature was observed in the 1.75 hydrate of ciprofloxacin salicylate n Table\u00a01. The chaA\u22efO1 hydrogen bonds (Table\u00a02a). Inter\u00adestingly, compound (II)b).The supra\u00admolecular features of compound (II)s Table\u00a02, formings Table\u00a02a. Interet al., 2019et al., 2020et al., 2020et al., 2020et al., 2014et al., 2016Several crystal structures of ciprofloxacin salts with benzoic acid derivatives have been reported, including salts with salicylic acid Uiso) were fixed to 1.2Ueq of the parent carbon or nitro\u00adgen atom and 1.5Ueq for hydroxyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022001177/dx2042sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989022001177/dx2042Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989022001177/dx2042IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989022001177/dx2042Isup4.cmlSupporting information file. DOI: 2098049, 2098403CCDC references: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N,N-di\u00admethyl\u00adformamide monosolvate was found to crystallize in the monoclinic space group P21/c. The crystal structure of this compound is stabilized by N\u2014H\u22efO and N\u2014H\u22efCl hydrogen bonds, as well as \u03c0\u2013\u03c0 stacking.9-Amino\u00adacridinium chloride N,N-di\u00admethyl\u00adformamide monosolvate, C13H11N2+Cl\u2212\u00b7C3H7NO, crystallizes in the monoclinic space group P21/c. The salt was crystallized from N,N-di\u00admethyl\u00adformamide. The asymmetric unit consists of two C13H11N2+Cl\u2212 formula units. The 9-amino\u00adacridinium (9-AA) mol\u00adecules are protonated with the proton on the N atom of the central ring. This N atom is connected to an N,N-di\u00admethyl\u00adformamide mol\u00adecule by a hydrogen bond. The H atoms of the amino groups create short contacts with two chloride ions. The 9-AA cations in adjacent layers are oriented in an anti\u00adparallel manner. The mol\u00adecules are linked via a network of multidirectional \u03c0\u2013\u03c0 inter\u00adactions between the 9-AA rings, and the whole lattice is additionally stabilized by electrostatic inter\u00adactions between ions.9-Amino\u00adacridinium chloride These bond lengths are characteristic for a C=N double bond that can originate from tautomerism of the cation, as shown on the scheme.The amino groups for two 9-amino\u00adacridine mol\u00adecules do not readily add a proton. The state of ionization is confirmed by both the H-atom positions (located from the difference map) and by the hydrogen bonding as shown in Table\u00a01et al., 1960The acridine moieties are nearly planar in the crystalline phase with atoms N2, C1, N1 and N5, C17 and N4 arranged almost linearly (N2\u22efC1\u2014N1 = 176\u00b0 and N5\u22efC17\u2014N4 = 180\u00b0). The dihedral angle between the two outer fused rings is 3.39\u2005(14)\u00b0 for the mol\u00adecule containing N2, while the corresponding angle in the mol\u00adecule containing N5 is 1.18\u2005(15)\u00b0. The second value is comparable with that found for acridine , which stack perpendicularly to the c axis. There are two types of 9-AA fused rings in the crystal structure, which results in the propagation of layers in a zigzag manner along b-axis direction .The packing of the mol\u00adecules in the crystal is illustrated in Fig.\u00a02on Fig.\u00a02.A, attached to N2 and N5, form hydrogen bonds to N,N-di\u00admethyl\u00adformamide atoms, O1 and O2, with d(N\u22efO) = 2.723\u2005(5)\u20132.740\u2005(5)\u2005\u00c5, N\u2014H\u22efO = 175-176\u00b0. The chloride ions are linked via N\u2014H\u22efCl hydrogen bonds , forming di\u00admers = 3.608\u2005(5)\u20133.688\u2005(4)\u2005\u00c5 and C\u2014H\u22efCl = 163-172\u00b0] to 4.2236\u2005(3)\u2005\u00c5. On the other hand, only the two aromatic rings of the acridine B mol\u00adecules participate in \u03c0\u2013\u03c0 inter\u00adactions, with adjacent acridine skeletons rotated in-plane with respect to one another. The centroid\u2013centroid distances vary from 3.6514\u2005(3) to 4.7445\u2005(5)\u2005\u00c5.Adjacent acridine skeletons are linked nt Fig.\u00a03. All of et al., 2016et al., 1974bP21/c space group with an 9-AA cation and a 3-chloro\u00adbenzoate anion in the asymmetric unit and the crystal structure features N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions. Inversely oriented cations and anions form a tetra\u00admer; these ions are linked via N(amino)\u2014H\u22efO (carb\u00adoxy) hydrogen bonds, forming a ring motif. 9-Amino\u00adacridinium 3-chloro\u00adbenzoate was dissolved in N,N-di\u00admethyl\u00adformamide (4\u2005ml) under heating at 418\u2005K until the 9-AA\u00b7HCl had fully dissolved. The solution was left to cool to 280\u2005K. Single crystals were obtained after 2 days.9-Amino\u00adacridinium hydro\u00adchloride = 1.2Ueq(C) for aromatic hydrogens and the C\u2014H group and C\u2014H = 0.96\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for the CH3 group. A rotating model was used for the methyl group.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021011816/dx2038sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021011816/dx2038Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021011816/dx2038Isup3.cmlSupporting information file. DOI: 2120699CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "This tiara-shaped hexa\u00admer crystallizing in the triclinic space group P4 centers, which are inter\u00adconnected through twelve \u03bc2-bridging benzyl thiol\u00adate groups. The Pd\u22efPd contacts range from 3.0892\u2005(2) to 3.1609\u2005(2)\u2005\u00c5 and can be considered as weakly bonding. The unit cell of Pd6 contains also a co-crystallized anthracene-9,10-dione mol\u00adecule.The first title compound, C R)(H)\u2013S unit can either be synthesized by nucleophilic substitution of geminal dihalides X\u2013C(R)(H)\u2013X by thiol\u00adates RS\u2212 \u00admeth\u00adyl]anthracenes with AgI salts 2], ] compounds is dominated by chelate complexes in which open-chain di\u00adthio\u00adether or macrocyclic polythio\u00adether ligands form five- or six-membered rings such as ] ], in which the thia\u00admacrocycle forms a strained four-membered chelate ring 4][ClO4]2, the strained chelate complexes [M(PhSCH2SPh)4](ClO4)2 are formed bis\u00ad(benzyl\u00adsulfane)-S,S\u2032)] was not formed according to the NMR data. Instead, a crystallographic study of a yellow\u2013orange crystal revealed the formation of a cyclic hexa\u00adnuclear thiol\u00adate-bridged cluster [Pd6(\u03bc2-SCH2Ph)12], Pd6. It is well known that thio\u00adacetals can be cleaved by soft HgII ions yielding aldehydes or other oxygenated products. One example is the HgIII-promoted deprotection of 3,5-bis\u00adBODIPYs, in which cleavage of a di\u00adthio\u00adacetal function to aldehyde groups occurs (benz\u00adyl)sulfane with ceric ammonium nitrate 2] with 2.1 equivalents of benzyl mercaptan in CH2Cl2 solution. However, the isolation of Pd6 was hampered by the co-crystallization of important amounts of the eight-membered cluster Pd8 [Pd8(\u03bc2-SCH2Ph)16], having a structure similar to that of [Pd8(\u03bc2-SPr)16] SBz] (SUNMAQ) and 2-[bis\u00ad(benzyl\u00adsulfan\u00adyl)meth\u00adyl]-6-meth\u00adoxy\u00adphenol [107.76\u2005(10)\u00b0], but considerably less than in [BzSCH2SBz] [117.33\u2005(7)\u00b0]. The phenyl rings of the benzyl groups and that of the anthracene unit form dihedral angles of 49.21\u2005(4) and 58.79\u2005(5)\u00b0.Compound L1 with [PdCl2(PhCN)2] shown in Fig.\u00a036(\u03bc2-SPr)12] 12] 12] 12] 12] 12] has been reported 12] is that of the phenyl\u00adethane\u00adthiol\u00adate-decorated nanocluster [Pd6(\u03bc2-SCH2CH2Ph)12] to 3.1609\u2005(2)\u2005\u00c5. The mean Pd\u22efPd separation of 3.1213\u2005(2)\u2005\u00c5 is quite similar to that of the other derivatives and may be considered as weakly bonding \u2005\u00c5 is close to those of the other [Pd6(\u03bc2-SR)12] analogues. The S\u2014Pd\u2014S bridge angles vary within the range 81.033\u2005(16)\u201399.246\u2005(16)\u00b0. The twelve sulfur atoms form two S6 hexa\u00adgons parallel to the central Pd6 ring from both sides, conferring finally a tiara-like shape to the Pd6S12 scaffold.The core of Note that the crystal structure of anthracene-9,10-dione has already been the object of several crystallographic studies and is therefore not commented herein distance of 2.519\u2005(18)\u2005\u00c5, the C14\u2014H14\u22efC24 distance of 2.741\u2005(18)\u2005\u00c5 and the C1\u2014H1B\u22efC9 distance of 2.847\u2005(16)\u2005\u00c5 are short enough to be considered as weak inter\u00admolecular inter\u00adactions . The red spots on the surface indicate the close contacts to adjacent mol\u00adecules. There are three areas of red spots which can be classified as C\u2014H\u22ef\u03c0 inter\u00adactions. The first and most important inter\u00adaction is the C\u2014H\u22ef\u03c0 contact of one of the phenyl\u00admethane\u00adthiol\u00adate substituents to the anthracene scaffold of a neighboring mol\u00adecule (C14\u2014H14\u22efC24). Furthermore, there are significant inter\u00adactions of the anthracene unit to an adjacent anthracene unit (C21\u2014H21\u22efC16/17/29). Then, there is also a weak C\u2014H\u22ef\u03c0 contact of two phenyl\u00admethane\u00adthiol\u00adate substituents (C1\u2014H1B\u22efC9). The contributions of the different types of inter\u00admolecular inter\u00adactions are shown in the two-dimensional fingerprint plots in Fig.\u00a08b and 6c illustrate the Hirshfeld surface mapped over the shape-index and the curvedness. The shape-index shows large red regions of concave curvature for the anthracene motif, whereas the C\u2014H-donors shows opposite curvature.A Hirshfeld surface analysis methyl-3,5-di\u00adnitro\u00adthio\u00adbenzoate 2 revealed only three similar structures, namely 2,6,10,14,19,24-hexa-p-benz-4,8,12,16,17,21,22,26-octa\u00adthia\u00adtri\u00adcyclo\u00adhexa\u00adcosa\u00adphane benzene clathrate 2(PMe3)2] 2ethane)] 2propane)] 2(bis\u00ad(di\u00adphenyl\u00adphosphino)methane)Cl2] 12] clusters have found applications as precursors for the preparation of monodisperse PdS nanoparticles 12] mol\u00adecules are inter\u00adconnected in the solid state by hydrogen bonds through the hy\u00addroxy groups of the thiol\u00adate ligands, thus generating an infinite three-dimensional supra\u00admolecular network 12] has been prepared and probed as a macrocyclic host to include an AgI ion as guest . Crystals suitable for single-crystal X-ray crystallography were grown by slow diffusion of hexane into a di\u00adchloro\u00admethane solution of L1, m.p. 438\u2013440\u2005K. 1H NMR : 9.03 , 8.39 , 8.00 , 7.95 , 7.55\u20137.47 , ddd , 7.28\u20137.22 , 7.14\u20137.09 , 6.91 , 5.94 , 3.79 , 3.55 . 13C{1H} NMR 138.34 (C16), 132.50 (C17), 131.46 (Cq), 131.36 (Cq), 130.28 (Cq), 129.58 (CHAr), 129.56 (C21), 129.47 (C25), 129.13 (Cq), 128.96 (CHAr), 128.84 (C23), 127.75 (C18), 127.53 (CHAr), 126.63 (C26), 125.61 (C19), 125.12 (C20), 124.91 (C27), 122.99 (C28), 45.02 (S2CH), 37.89 (SCH2). IR (ATR) cm \u22121: 3050 and 3025 (C\u2014H Ar), 2998, 2948 and 2906 , 1589, 1519 (C=C), 696 (C\u2014S).9-Anthracenecarboxaldehyde and benzyl mercaptan were suspended in conc. HCl (2\u2005ml) and allowed to stir at room temperature. After 2\u2005h, the reaction mixture was neutralized with aqueous NaHCO2(PhCN)2Reaction of L1 with PdCl: L1 and PdCl2(PhCN)2 were dissolved in 5\u2005ml of di\u00adchloro\u00admethane and allowed to stir at room temperature for 30 minutes. During the reaction, a red solution was obtained, which was kept in refrigerator overnight yielding yellow crystals of 9-anthraldehyde along with yellow\u2013orange co-crystals of the cluster, Pd6. 1H NMR ): 8.92\u20136.86 , 3.61 , 3.58 .Uiso(H) = 1.2Ueq(C). Hydrogen atoms H1B, H14 and H21 for L1 were located in the difference-Fourier map and refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021006113/hb7976sup1.cifCrystal structure: contains datablock(s) mo_b0159_0m, mo_b0283_0m, New_Global_Publ_Block. DOI: 10.1107/S2056989021006113/hb7976mo_b0159_0msup2.hklStructure factors: contains datablock(s) mo_b0159_0m. DOI: 10.1107/S2056989021006113/hb7976mo_b0283_0msup3.hklStructure factors: contains datablock(s) mo_b0283_0m. DOI: 2089413, 2089412CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Preoperative pulmonary function testing is mandatory for non-small cell lung cancer (NSCLC) surgery. The predicted postoperative FEV1 (ppoFEV1) is used for further risk stratification. We compared the ppoFEV1 with the postoperative FEV1 (postFEV1) in order to improve the calculation of the ppoFEV1.87 patients voluntarily received an FEV1 assessment 1\u00a0year after surgery. ppoFEV1 was calculated according to the Brunelli calculation. Baseline characteristics and surgical procedure were compared in a uni- and multivariate analysis between different accuracy levels of the ppoFEV1. Parameters which remained significant in the multinominal regression analysis were evaluated for a modification of the ppoFEV1 calculation.p\u2009=\u20090.01), packyears , younger age , and patients undergoing pneumectomy . For the customized ppoFEV1 we excluded pneumonectomies. For patients\u2009<\u200960\u00a0years, an additional lung segment was added to the calculation. ppoFEV1\u2009=\u2009preFEV1\u2009\u00d7\u2009Independent factors for a more inaccurate ppoFEV1 were preoperative active smoking (odds ratio (OR) 4.1, confidence interval (CI) 3.6\u20136.41; We were able to enhance the predictability of the ppoFEV1 with modifications. The modified ppoFEV1 (1.828\u00a0l\u2009\u00b1\u20090.479\u00a0l) closely approximates the postFEV1 of 1.823\u00a0l\u2009\u00b1\u20090.476\u00a0l, (0.27%) while the original ppoFEV1 calculation is at 1.78\u00a0l\u2009\u00b1\u20090.53 (2.19%). However, if patients require pneumectomy, more complex techniques to determine the ppoFEV1 should be included to stratify risk.The online version contains supplementary material available at 10.1007/s00408-021-00464-4. Anatomical lung resection is the gold standard for the treatment of early-stage non-small cell lung cancer (NSCLC) . PulmonaAll relevant patient data were taken from the electronic hospital information system of our institute. 87 patients voluntarily presented to our out-patient department 1 year after surgery for a routine surgical check-up which included a pulmonary function test. We included all patients who received a postFEV1 in our analysis.The ppoFEV1 is calculated based on the preoperative FEV1 (preFEV1), the number of functional lung segments resected (y), and the total number of functional segments available at time of resection (z). PpoFEV1\u2009=\u2009preFEV1\u2009\u00d7\u20091 \u2212 entclass1pt{minimamentclasspt{minimaAfter calculating the ppoFEV1, we compared our results with the postFEV1 1\u00a0year after surgery.We determined accuracy levels and classified the deviation of the ppoFEV1 in relation to the postFEV1. In order to reflect the greatest differences, we defined the accuracy level of\u2009\u00b1\u20092% as most accurate, the accuracy level of\u2009\u00b1\u200910% as moderately accurate, and the accuracy level of\u2009\u00b1\u2009\u2009>\u200910% as inaccurate. We performed a subgroup analysis excluding the ppoFEV1\u2009\u00b1\u20092% from the ppoFEV1\u2009\u00b1\u200910%, in order to prevent counting ppoFEV1\u2009\u00b1\u20092% patients twice. These patients were defined as ppoFEV1\u2009\u00b1\u200910%\u2009>\u2009\u2009\u00b1\u20092%. We analyzed baseline characteristics and surgical procedures which eventually resulted in the most accurate- or inaccurate ppoFEV1 with univariate and multivariate analyses.\u03c72 or Fisher\u2019s exact test. Continuous parameters were expressed as mean \u00b1 standard deviation (SD) and were analyzed by an unpaired Student t test. p-value <0.05 was considered statistically significant. Statistical analysis was performed using the SPSS statistical software package .In cases where univariate analysis showed significant differences, we performed a multinominal regression analysis for further evaluation. Multinomial differences are described by odds ratio (OR) and 95% confidence interval (CI). Categorical variables were analyzed using Pearson\u2019s Significant multinominal parameters were included in our customizing process. The primary objective of this customization was to determine baseline characteristics or surgical procedures which could be included in the calculation in order to improve the ppoFEV1.The customizing process is explained in the results part. The formulas calculated by us are the following.For patients\u2009<\u200960\u00a0years, an additional lung segment is added to the calculation ppoFEV1\u2009=\u2009preFEV1\u2009\u00d7\u2009A total of 464 patients underwent anatomical pulmonary resections since 2012 at our institution. 87 (18.8%) patients presented voluntarily for a postoperative check-up and a redo assessment of their pulmonary function 1 year after surgery. We classified patients into categories according to how accurately the ppoFEV1 predicted the postFEV1. The ppoFEV1 of 79% patients showed moderate accuracy of\u2009\u00b1\u200910%. 24% of patients were most accurately predicted with a\u2009\u00b1\u20092% deviation. Calculated values exceeded\u2009>\u2009\u2009\u00b1\u200910% in 21% of patients. 51 (45%) patients showed a ppoFEV1\u2009\u00b1\u200910%\u2009>\u2009\u2009\u00b1\u20092% accuracy level. The mean preFEV1 of the cohort was 2.34 l (l)\u2009\u00b1\u20090.61\u00a0l and the mean postFEV1 was 1.82\u00a0l\u2009\u00b1\u20090.47\u00a0l, respectively. Calculation of the ppoFEV1 yielded 1.78\u00a0l\u2009\u00b1\u20090.53\u00a0l (2.19%).p\u2009=\u20090.01).The baseline characteristics according to the predictive accuracy level are presented in Tables p\u2009=\u20090.05). The preoperative DLCO% (preDLCO%) of patients with ppoFEV1\u2009\u00b1\u20092% showed a significantly higher value (87.4\u2009\u00b1\u200917.7) than in ppoFEV1\u2009\u00b1\u200910\u2009>\u2009\u2009\u00b1\u20092% patients (77.3\u2009\u00b1\u200919.9) (p\u2009=\u20090.05). The postDLCO however did not differ between\u2009\u00b1\u20092% and\u2009\u00b1\u200910\u2009>\u2009\u2009\u00b1\u20092%. 74.7% of patients were active smokers at the time of surgery (active smoking: smoking in 3\u00a0months prior to surgery). Active smoking did not differ between ppoFEV1\u2009\u00b1\u20092% and ppoFEV1\u2009\u00b1\u200910%\u2009>\u2009\u2009\u00b1\u20092%. 100% of ppoFEV1\u2009>\u2009\u2009\u00b1\u200910% patients were active smokers compared to 71.0% of ppoFEV1\u2009\u00b1\u200910% . Patients with ppoFEV1\u2009>\u2009\u2009\u00b1\u200910% smoked significantly more than ppoFEV1\u2009\u00b1\u200910% patients (61.4\u2009\u00b1\u200919.1 vs. 41.2\u2009\u00b1\u200924.5 p\u2009=\u20090.002). Heavy smoking is defined as\u2009>\u200930 packyears [p\u2009=\u20090.001).The distribution of age in relation of the ppoFEV1 is shown in Fig.\u00a0 77.3\u2009\u00b1\u20091.9 . However, younger age , active smoking , packyears , and heavy smoking (>\u200930 packyears) were independent factors for a more inaccurate ppoFEV1.We carried out a multinominal regression analysis for further evaluation of significant results from the univariate analysis (included in Tables p\u2009=\u20090.05). The right lower lobe was resected less often in patients with ppoFEV1\u2009\u00b1\u20092% . Patients with ppoFEV1\u2009\u00b1\u20092% never underwent pneumectomy (0 vs. 3.0%) or bilobectomy (0% vs. 6.0%). The most inaccurate FEV1\u2009>\u2009\u2009\u00b1\u200910% was significantly more frequent in patients undergoing pneumectomy . Pneumectomy on the other was an independent factor for ppoFEV1\u2009>\u2009\u2009\u00b1\u200915% in the multivariate analysis \u2009<\u200960\u00a0years and (3) active heavy smokers we used the original calculation as the modifications canceled each other.To improve the accuracy of the ppoFEV1, we tried to keep the method as simple as possible.In the following section we give two computational examples with the preFEV1 of two patients explaining our customized ppoFEV1.ppoFEV1\u2009=\u2009preFEV1 x preFEV1\u2009=\u20092.67\u00a0l; postFEV1\u2009=\u20092.04\u00a0l; ppoFEV1 2.25\u00a0lCalculation method: 2.67\u00a0l\u2009\u00d7\u20091\u2009\u2212\u2009Customized method: 2.67\u00a0l\u2009\u00d7\u20091\u2009\u2212\u2009Patient undergoing an upper right lobe resection\u2009<\u200960\u00a0years:ppoFEV1\u2009=\u2009PreFEV1 \u00d7 Calculation method: 1.77 \u00d7 Customized method: 1.77 \u00d7 Patient undergoing a lower left lobe resection heavy smoker:The mean postFEV1 was 1.823\u00a0l\u2009\u00b1\u20090.476\u00a0l. By customizing the ppoFEV1 the calculation was more accurate (1.828\u00a0l\u2009\u00b1\u20090.479\u00a0l) (0.27%) compared to the original ppoFEV1 (1.78\u00a0l\u2009\u00b1\u20090.53\u00a0l) (2.19%). In addition, customization also led to a redistribution of the accuracy levels we classified Below is the link to the electronic supplementary material."} +{"text": "The manganese(II) centers of each structure are six-coordinate with a distorted octa\u00adhedral geometry. Although the bis\u00ad(quinolin-2-ylmeth\u00adyl)ethanamine ligands differ only by a methyl group, the structure of one complex is dimeric with bridging acetate ligands and exhibits a 4O:O\u2032-bis\u00ad{manganese(II)} bis\u00ad(tetra\u00adphen\u00adyl\u00adborate) di\u00adchloro\u00admethane 1.45-solvate, [Mn2(C23O2)2(C23H23N3O)2](C24H20B)\u00b71.45CH2Cl2 or [Mn(DQMEA)(\u03bc-OAc)2Mn(DQMEA)](BPh4)2\u00b71.45CH2Cl2 or [1](BPh4)2\u00b71.45CH2Cl2, and (acetato-\u03baO)(methanol-\u03baO)manganese(II) tetra\u00adphenyl\u00adborate methanol monosolvate, [Mn(CH3COO)(C22H21N3O)(CH3OH)](C24H20B)\u00b7CH3OH or [Mn(DQEA)(OAc)(CH3OH)]BPh4\u00b7CH3OH or [2]BPh4\u00b7CH3OH, by single-crystal X-ray diffraction reveal distinct differences in the geometry of coordination of the tripodal DQEA and DQMEA ligands to MnII ions. In the asymmetric unit, compound [1](BPh4)2\u00b7(CH2Cl2)1.45 crystallizes as a dimer in which each manganese(II) center is coordinated by the central amine nitro\u00adgen, the nitro\u00adgen atom of each quinoline group, and the meth\u00adoxy-oxygen of the tetra\u00addentate DQMEA ligand, and two bridging-acetate oxygen atoms. The symmetric MnII centers have a distorted, octa\u00adhedral geometry in which the quinoline nitro\u00adgen atoms are trans to each other resulting in co-planarity of the quinoline rings. For each MnII center, a coordinated acetate oxygen participates in C\u2014H\u22efO hydrogen-bonding inter\u00adactions with the two quinolyl moieties, further stabilizing the trans structure. Within the crystal, weak \u03c0\u2013\u03c0 stacking inter\u00adactions and inter\u00admolecular cation\u2013anion inter\u00adactions stabilize the crystal packing. In the asymmetric unit, compound [2]BPh4\u00b7CH3OH crystallizes as a monomer in which the manganese(II) ion is coordinated to the central nitro\u00adgen, the nitro\u00adgen atom of each quinoline group, and the alcohol oxygen of the tetra\u00addentate DQEA ligand, an oxygen atom of OAc, and the oxygen atom of a methanol ligand. The geometry of the MnII center in [2]BPh4\u00b7CH3OH is also a distorted octa\u00adhedron, but the quinoline nitro\u00adgen atoms are cis to each other in this structure. Hydrogen bonding between the acetate oxygen atoms and hydroxyl (O\u2014H\u22efO) and quinolyl (C\u2014H\u22efO and N\u2014H\u22efO) moieties of the DQEA ligand stabilize the complex in this cis configuration. Within the crystal, dimerization of complexes occurs by the formation of a pair of inter\u00admolecular O3\u2014H3\u22efO2 hydrogen bonds between the coordinated hydroxyl oxygen of the DQEA ligand of one complex and an acetate oxygen of another. Additional hydrogen-bonding and inter\u00admolecular cation\u2013anion inter\u00adactions contribute to the crystal packing.Structural analyses of the compounds di-\u03bc-acetato-\u03ba These compounds are prepared in a two-step reaction (see reaction scheme) in which mangan\u00adese(II) acetate is reacted with either DQMEA or DQEA in methanol, followed by anion exchange with sodium tetra\u00adphenyl\u00adborate. The resulting complexes demonstrate how minor alterations in ligand structure can result in significant differences in the complex structure.We have recently begun to study Mn[1](BPh4)2\u00b7(CH2Cl2)1.45 crystallizes in the triclinic space group P2Mn(DQMEA)]2+ cation, [1] in the same octa\u00adhedral plane and the meth\u00adoxy oxygen (O1) located perpendicular to this nitro\u00adgen plane. This configuration of the DQMEA ligand results in the quinoline groups binding MnIItrans to each other, and in coplanarity of their rings. Hydrogen-bonding inter\u00adactions between quinolyl hydrogens and an acetate oxygen, C\u2014H\u22efO, further stabilize this trans configuration and 75.56\u2005(5)\u00b0, respectively, which are significantly reduced from 90\u00b0 (Table\u00a01trans N1\u2014Mn1\u2014N3 angle of 148.35\u2005(5)\u00b0. Likewise, the bond angle formed by cis coordination of the meth\u00adoxy oxygen of DQMEA and central nitro\u00adgen, N2\u2014Mn1\u2014O1 is 75.32\u2005(5)\u00b0. The remaining trans bond angles, O2\u2014Mn1\u2014N2 and O31\u2014Mn1\u2014O1 are 157.89\u2005(5) and 163.58\u2005(5)\u00b0, respectively. The Mn\u2014O and Mn\u2014N bond lengths for the neutral DQMEA ligand fall in the range 2.27\u20132.36\u2005\u00c5, which is typical of manganese(II) complexes and 2.0908\u2005(14)\u2005\u00c5, are significantly shorter.Compound P Fig.\u00a01. The str1] Fig.\u00a02 balancedn Table\u00a03. Oxygens\u00b0 Table\u00a01. This re[2]BPh4\u00b7CH3OH crystallizes in the monoclinic space group P21/c. The structure of this compound consists of the [Mn(DQEA)(OAc)(CH3OH)]+ monocation, [2], tetra\u00adphenyl borate counter-ion, and a methanol solvent mol\u00adecule and 73.81\u2005(5)\u00b0, respectively \u00b0. The remaining trans bond angles, O1\u2014Mn1\u2014N1 and N2\u2014Mn1\u2014O4 are 175.54\u2005(6) and 161.38\u2005(6)\u00b0, respectively.The compound le Fig.\u00a03. The MnIy Table\u00a02. The alccis coordination of DQEA to Mn(II) in [2] may result from a hydrogen-bonding network involving the alcohol and quinolyl groups of DQEA and the acetate ligand, O\u2014H\u22efO and C\u2014H\u22efO 2\u00b7(CH2Cl2)1.45, no classical inter\u00admolecular hydrogen bonding inter\u00adactions were found. The crystal packing of a quinoline group . In addition, a network of weak C\u2014H\u22ef\u03c0 inter\u00admolecular cation\u2013anion inter\u00adactions inter\u00admolecular cation\u2013anion inter\u00adactions compounds described herein have not been reported previously. We have previously reported the structure of a mononuclear copper(II) complex with DQMEA ethanamine (DQMEA). In a 250\u2005ml round-bottom flask, 5\u2005g (23\u2005mmol) of 2-chlormethyl\u00adquinoline hydro\u00adchloride was dissolved in 10\u2005ml of H2O and cooled to 273\u2005K in an ice bath. A solution of 1.9\u2005g (47\u2005mmol) of NaOH in 10\u2005ml of H2O was added dropwise with stirring. Following this, a solution of 0.9\u2005g (12\u2005mmol) of 2-meth\u00adoxy\u00adethyl\u00adamine in 10\u2005ml of CH2Cl2 was added. The reaction mixture was then removed from the ice bath, and brought to reflux for 7 days. The mixture was then cooled to room temperature, and the CH2Cl2 layer was separated, washed twice with brine, and dried over anhydrous sodium sulfate. The solution was then filtered, and the filtrate was chromatographed on alumina eluting with 20:1 CH2Cl2/methanol. Fractions were collected that produced a single spot by TLC on alumina plates with an RF value of 0.33. Rotary evaporation of these fractions gave 2.4\u2005g (58%) of a light-yellow solid. 1H NMR \u03b4 2.87 , 3.30 , 3.54 , 4.06 , 7.48 , 7.65 , 7.75 , 8.01 , 8.10 .2-Hy\u00addroxy-N,N-bis\u00ad(quinolin-2-ylmeth\u00adyl)ethanamine (DQEA). In a 100\u2005ml round-bottom flask, 2.5\u2005g (12\u2005mmol) of 2-chlormethyl\u00adquinoline hydro\u00adchloride was dissolved in 10\u2005ml of H2O and cooled to 273\u2005K in an ice bath. A solution of 0.95\u2005g (24\u2005mmol) of NaOH in 10\u2005ml of H2O was added dropwise with stirring. Following this, a solution of 0.36\u2005g (6.0\u2005mmol) of ethano\u00adlamine in 10\u2005ml of CH2Cl2 was added. The reaction mixture was then removed from the ice bath, and brought to reflux for 7 days. The mixture was then cooled to room temperature, and the CH2Cl2 layer was separated, washed twice with brine, and dried over anhydrous sodium sulfate. The solution was then filtered, and the filtrate was chromatographed on alumina eluting with 100:1 CH2Cl2/methanol. Fractions were collected that produced a single spot by TLC on alumina plates with an RF value of 0.33. Rotary evaporation of these fractions gave 0.70\u2005g (20%) of a light-yellow solid. 1H NMR \u03b4 3.02 , 3.54 , 4.17 , 7.51 , 7.74 , 8.07 .2Mn(DQMEA)](BPh4)2BPh4\u00b7CH3OH(BPh4)2\u00b7(CH2Cl2)1.45 and [2]BPh4\u00b7CH3OH are summarized in Table\u00a05[1](BPh4)2\u00b7(CH2Cl2)1.45, all H atoms were positioned geometrically and refined using a riding model: C\u2014H = 0.93\u20130.99\u2005\u00c5, with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl). Idealized methyl groups were refined as rotating groups. A solvate methyl\u00adene chloride mol\u00adecule was refined as threefold disordered. All C\u2014Cl bond distances were restrained to be the same within a standard deviation of 0.02\u2005\u00c5. Uij components of ADPs were restrained to be similar to each other . Occupancies were not constrained to unity and refined to 0.401\u2005(3), 0.234\u2005(4) and 0.090\u2005(4). In [2]BPh4\u00b7CH3OH, the ethanol group of C21, C22 and O3 was found to be disordered. Bond distances and angles of major and minor moiety were restrained to be similar to each other . Uij components of ADPs were restrained to be similar to each other . The hy\u00addroxy H atoms were located in a difference-Fourier map and refined with the distance restraint O\u2014H = 0.8\u2005(2)\u2005\u00c5 and with Uiso(H) = 1.5Ueq(O). C-bound H atoms were positioned geometrically and refined as riding: C\u2014H = 0.95\u20130.99\u2005\u00c5 with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl). Idealized methyl groups were refined as rotating groups. An idealized tetra\u00adhedral OH group was also refined as a rotating group: O1S(H1S).Crystal data, data collection and structure refinement details for 10.1107/S2056989021009786/zl5024sup1.cifCrystal structure: contains datablock(s) 1, 2. DOI: 10.1107/S2056989021009786/zl50241sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989021009786/zl50242sup3.hklStructure factors: contains datablock(s) 2. DOI: 2110882, 2110881CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Nature Communications 10.1038/s41467-021-21863-4, published online 12 March 2021.Correction to: 2\u03c0\u22121\u2212\u03c3m2\u03c4\u22121 . The correct form of the equation is: D\u2009=\u2009(M2\u03c0\u22121\u2212\u03c3m2)\u03c4\u22121.The original version of this Article contained an error in the Methods, section \u2018Single-particle tracking photoactivated localization microscopy (sptPALM)\u2019, where an equation incorrectly read: D\u2009=\u2009MThis has been corrected in both the PDF and HTML versions of the Article."} +{"text": "Embryo implantation into the uterus is a crucial step for human reproduction. A hypothesis has been proposed that the molecular circuit invented by trophoblasts for invasive embryo implantation during evolution might be misused by cancer cells to promote malignancy. Unfortunately, our current understanding of the molecular mechanism underlying embryo implantation is far from complete.Here we used the mouse as an animal model and generated a single-cell transcriptomic atlas of the embryo implantation site of mouse uterus at the invasion phase of embryo implantation on gestational day 6. We revealed 23 distinct cell clusters, including 5 stromal cell clusters, 2 epithelial cell clusters, 1 smooth muscle cell cluster, 2 pericyte clusters, 4 endothelial cell clusters, and 9 immune cell clusters. Through data analysis, we identified differentially expression changes in all uterine cell types upon embryo implantation. By integrated with single-cell RNA-seq data from E5.5 embryos, we predicted cell\u2013cell crosstalk between trophoblasts and uterine cell types.Our study provides a valuable resource for understanding of the molecular mechanism of embryo implantation.The online version contains supplementary material available at 10.1186/s13578-022-00749-y. Embryo implantation in humans is interstitial. It consists of the following three phases: embryo apposition, attachment, and invasion. Immediately after apposition and attachment to endometrial epithelium, the blastocyst penetrates through the epithelium, followed by the basal lamina, and invades into the stroma . It has Due to ethical restrictions and experimental difficulties, in vivo analysis of embryo implantation heavily relies on mice . SlightlIn the present study, by using the-state-of-the-art single-cell RNA-seq approach , we resoAdult CD-1 mice of the SPF grade were used in this study. All mice were caged under light-controlled conditions (14\u00a0h/10\u00a0h light/dark cycles) with free access to regular food and water. Female mice were mated with fertile males and success of mating was confirmed the next morning by the presence of a vaginal plug. The day of the vaginal plug was denoted as gestation day 1 (GD1). On GD6, the implantation sites and inter-implantation sites (served as a control) were collected separately. All animal procedures were approved by the Institutional Animal Care and Use Committee of South China Agricultural University .The total RNAs from uterine tissues were extracted with the TRIzol reagent (Invitrogen). RNA-seq libraries were generated by using the TruSeq RNA sample preparation kit (Illumina) and sequenced on an Illumina HiSeq 2500 system. Using UCSC mm10 mouse genome as reference, raw data were analyzed using TopHat v2.0.4 and CuffSingle-cell suspension was prepared as described previously , 17. BriThe final concentration of single-cell suspension was adjusted to 1000 cells/\u03bcl and a volume of 15\u00a0\u00b5l was loaded into one channel of the ChromiumTM Single Cell B Chip , aiming at recovering 8000\u201310,000 cells. The Chromium Single Cell 3' Library & Gel Bead Kit v3 was used for single-cell bar-coding, cDNA synthesis and library preparation, following the manufacturer's instructions provided as the Single Cell 3' Reagent Kits User Guide Version 3. Library sequencing was performed on an Illumina novaseq 6000 system configured with the paired-end 150-bp protocol at a sequencing depth of approximately 400 million reads.Raw data bcl files from the Illumina NovaSeq 6000 platform were converted to fastq files using the bcl2fastq tool (v2.19.0.316). These fastq files were aligned to the mm10 mouse reference genome by using the CellRanger software . The resulting gene counts matrix was analyzed with the R package Seurat (v3.1.3) . Cell wiMonocle2 package v2.18.0 was usedGene ontology (GO) analysis was performed as described previously . GO termPathway enrichment analysis was conducted by using the Metascape v7.4 online tools . The sigThe CellChat v1.1.0 software was usedTo create a cell-type resolved map of mouse uterus at the invasion phase of embryo implantation, we performed single-cell RNA-seq analysis Fig.\u00a0A. The imUnsupervised clustering analysis revealed 23 distinct cell clusters for all cells from IIS and IS combined [+Nkg7\u2212Cd3e+Mki67+), T cells [+Cd79a+Ms4a1+) [+Adgre1+) [+Adgre1+Mki67+), dendritic cells [+Itgax+Mki67+) and plasmacytoid dendritic cells [Hormone-responsive cells included epithelial cells expressing Epcam and Krt19 Fig.\u00a02CC2C, stro7+Cd3e\u2212) , prolife7\u2212Cd3e+) , B cells+Ms4a1+) , macrophAdgre1+) , prolife+Itgax+) , prolifeiglech+) .Finally, we aimed to discover novel markers for each cell type. We selected genes that were expressed significantly higher in the cell type of interest than the other cell types by Wilcoxon rank sum test. A complete list of these marker genes was presented in Additional file \u201311), developmental processes (P\u2009=\u20091.10\u2009\u00d7\u200910\u20134), cell organization and biogenesis (P\u2009=\u20093.27\u2009\u00d7\u200910\u20134), stress response (P\u2009=\u20094.88\u2009\u00d7\u200910\u20134) and protein metabolism (P\u2009=\u20093.87\u2009\u00d7\u200910\u20132). Gene set 2 with 352 genes were decreased in S3 compared to its intermediate S2p. These genes were enriched in DNA metabolism (P\u2009=\u20091.00\u2009\u00d7\u200910\u201311), cell cycle and proliferation (P\u2009=\u20091.00\u2009\u00d7\u200910\u201311), cell organization and biogenesis (P\u2009=\u20092.59\u2009\u00d7\u200910\u201311) and protein metabolism (P\u2009=\u20093.97\u2009\u00d7\u200910\u20133). Gene set 3 of 300 genes were S2-specific. Based on GO, enriched terms were protein metabolism (P\u2009=\u20093.39\u2009\u00d7\u200910\u201311), developmental processes (P\u2009=\u20096.05\u2009\u00d7\u200910\u20138) and cell cycle & proliferation (P\u2009=\u20098.09\u2009\u00d7\u200910\u20133). Gene set 4 of 729 genes were unchanged or increased in S3 compared to its intermediate S2p. Enriched GO terms were protein metabolism (P\u2009=\u20092.49\u2009\u00d7\u200910\u20138), RNA metabolism (P\u2009=\u20093.23\u2009\u00d7\u200910\u20137), DNA metabolism (P\u2009=\u20094.42\u2009\u00d7\u200910\u20135), transport (P\u2009=\u20091.96\u2009\u00d7\u200910\u20133) cell organization and biogenesis (P\u2009=\u20092.90\u2009\u00d7\u200910\u20135), and cell cycle and proliferation (P\u2009=\u20094.91\u2009\u00d7\u200910\u20133).In our single-cell RNA-seq data, we identified 5 clusters of stromal cells: S1 , S1p , S2 , S2p and S3 . We selected signature genes for each cell cluster by using Wilcoxon rank sum test. After the removal of redundancy, we identified a total of 1784 signature genes were examined. We found that PDZ expressed pan-stromal cell markers Pgr, Esr1 and Hoxa11, as well as the superficial stromal cell marker Hand2. Additionally, PDZ expressed decidualization marker Wnt4, but not Prl8a2 . PDZ cea were exaTo further reveal the relationship between these 5 stromal cell clusters, pseudotime trajectory analysis was conducted. Cells were arranged in a pseudotime manner with a pedigree reconstruction algorithm for biological processes based on transcriptional similarity. We found 2 paths of interest: (1) primary decidual zone formation, i.e. S2-\u2009>\u2009S2p/S3; and (2) secondary decidual zone formation, i.e. S1-\u2009>\u2009S1p , PI3K-Akt signaling pathway (FDR\u2009=\u20091.00\u2009\u00d7\u200910\u201318), Regulation of actin cytoskeleton (FDR\u2009=\u20091.26\u2009\u00d7\u200910\u20139), Rap1 signaling pathway (FDR\u2009=\u20091.58\u2009\u00d7\u200910\u20136), Ras signaling pathway (FDR\u2009=\u20097.94\u2009\u00d7\u200910\u20135), MAPK signaling pathway (FDR\u2009=\u20091.26\u2009\u00d7\u200910\u20134), Phospholipase D signaling pathway (FDR\u2009=\u20097.94\u2009\u00d7\u200910\u20133) and Cytokine-cytokine receptor interaction (FDR\u2009=\u20093.98\u2009\u00d7\u200910\u20132) and TGC. We found a total of 20 ligand-receptor interaction pairs Fig.\u00a0F. Based FDR\u2009=\u20097.9\u2009\u00d7\u200910\u20135, 2 test was employed to assess the significance of difference between two groups. By using the criteria of P\u2009<\u20090.05 and fold change\u2009>\u20092, we found that the proportion of S1 was unchanged, whereas the proportion of S2 significantly decreased in IS compared to IIS. Meanwhile, S1p and S2p were almost exclusively detected in IS , PI3K-Akt signaling pathway (FDR\u2009=\u20092.00\u2009\u00d7\u200910\u20139), Regulation of actin cytoskeleton (FDR\u2009=\u20093.16\u2009\u00d7\u200910\u20139), Hippo signaling pathway (FDR\u2009=\u20093.98\u2009\u00d7\u200910\u20139), ECM-receptor interaction (FDR\u2009=\u20093.98\u2009\u00d7\u200910\u20138), Rap1 signaling pathway (FDR\u2009=\u20091.00\u2009\u00d7\u200910\u20136), MAPK (FDR\u2009=\u20093.16\u2009\u00d7\u200910\u20136), Wnt signaling pathway (FDR\u2009=\u20092.00\u2009\u00d7\u200910\u20135), Ras signaling pathway (FDR\u2009=\u20092.00\u2009\u00d7\u200910\u20135), mTOR signaling pathway (FDR\u2009=\u20092.51\u2009\u00d7\u200910\u20135), TGF-beta signaling pathway (FDR\u2009=\u20092.51\u2009\u00d7\u200910\u20135), Endocytosis (FDR\u2009=\u20096.31\u2009\u00d7\u200910\u20135) and Phagosome (FDR\u2009=\u20097.94\u2009\u00d7\u200910\u20133) on S1 and S2, we used the CellChat software to predict the ligand-receptor interactions. Only secreted factors from PDZ were considered. We found a total of 35 ligand-receptor interaction pairs Fig.\u00a0A. PathwaFDR\u2009=\u20093.1\u2009\u00d7\u200910\u20139, By using the criteria of P\u2009<\u20090.05 and fold change\u2009>\u20092, we found that the proportion of NKp, Mp and DCp were significantly increased at IS compared IIS Fig.\u00a0A. Using \u20137), PI3K-Akt signaling pathway (FDR\u2009=\u20091.26\u2009\u00d7\u200910\u20136), MAPK signaling pathway (FDR\u2009=\u20093.16\u2009\u00d7\u200910\u20134), Endocytosis (FDR\u2009=\u20093.98\u2009\u00d7\u200910\u20134), ECM-receptor interaction (FDR\u2009=\u20093.98\u2009\u00d7\u200910\u20134), TGF-beta signaling pathway (FDR\u2009=\u20093.98\u2009\u00d7\u200910\u20134), Rap1 signaling pathway (FDR\u2009=\u20097.94\u2009\u00d7\u200910\u20134), Hippo signaling pathway (FDR\u2009=\u20092.51\u2009\u00d7\u200910\u20133), Regulation of actin cytoskeleton (FDR\u2009=\u20097.94\u2009\u00d7\u200910\u20133), Toll-like receptor signaling pathway (FDR\u2009=\u20091.00\u2009\u00d7\u200910\u20132), Ras signaling pathway (FDR\u2009=\u20091.26\u2009\u00d7\u200910\u20132), Natural killer cell mediated cytotoxicity (FDR\u2009=\u20091.58\u2009\u00d7\u200910\u20132), Jak-STAT signaling pathway (FDR\u2009=\u20093.16\u2009\u00d7\u200910\u20132) and NOD-like receptor signaling pathway (FDR\u2009=\u20093.98\u2009\u00d7\u200910\u20132) and immune cells using the CellChat software. We found a total of 26 ligand-receptor interaction pairs Fig.\u00a0A. PathwaFDR\u2009=\u20093.9\u2009\u00d7\u200910\u20134, By using the criteria of P\u2009<\u20090.05 and fold change\u2009>\u20092, we found that the proportions of both VEC and LEC were significantly decreased in IS compared to IIS, which is in line with the fact that PDZ is avascular \u201344. Inte\u201323), Rap1 signaling pathway (FDR\u2009=\u20091.00\u2009\u00d7\u200910\u201316), Ras signaling pathway (FDR\u2009=\u20091.00\u2009\u00d7\u200910\u201314), Cytokine-cytokine receptor interaction (FDR\u2009=\u20091.00\u2009\u00d7\u200910\u201314), HIF-1 signaling pathway (FDR\u2009=\u20091.26\u2009\u00d7\u200910\u20137), Hippo signaling pathway (FDR\u2009=\u20091.00\u2009\u00d7\u200910\u20136), Endocytosis (FDR\u2009=\u20093.16\u2009\u00d7\u200910\u20136), Regulation of actin cytoskeleton (FDR\u2009=\u20096.31\u2009\u00d7\u200910\u20136), mTOR signaling pathway (FDR\u2009=\u20091.58\u2009\u00d7\u200910\u20135), MAPK signaling pathway (FDR\u2009=\u20091.58\u2009\u00d7\u200910\u20135), ECM-receptor interaction (FDR\u2009=\u20093.98\u2009\u00d7\u200910\u20134), TGF-beta signaling pathway (FDR\u2009=\u20093.98\u2009\u00d7\u200910\u20134), NOD-like receptor signaling pathway (FDR\u2009=\u20095.01\u2009\u00d7\u200910\u20133), Toll-like receptor signaling pathway (FDR\u2009=\u20091.26\u2009\u00d7\u200910\u20132), AMPK signaling pathway (FDR\u2009=\u20092.00\u2009\u00d7\u200910\u20132), Wnt signaling pathway (FDR\u2009=\u20093.16\u2009\u00d7\u200910\u20132) and Jak-STAT signaling pathway (FDR\u2009=\u20093.98\u2009\u00d7\u200910\u20132) and VEC/LEC. We found a total of 34 ligand-receptor interaction pairs Fig.\u00a0A. PathwaEmbryo implantation is a crucial step for human embryo implantation. Previously, we performed single-cell RNA analysis of the mouse uterus during the apposition phase and attachment phase of embryo implantation on gestational days (GD) 4\u20135 , 17. HerIn order to accommodate embryo implantation, an important change in mouse uterus is the formation of primary decidual zone (PDZ). The first sign of PDZ formation occurs on the afternoon of GD5. PDZ is fully established by GD6 \u201347. In tThe trophectoderm (TE) of mouse embryo is created by the end of the pre-implantation period. After implantation, TE develops into two different types, polar TE and mural TE. Polar TE differentiates into extraembryonic ectoderm (ExE) and ectoplacental cone (EPC), while mural TE differentiates into trophoblast giant cells (TGC). TGC cells are in closest contact to the uterus during embryo implantation . Thus, tBesides the emergence of PDZ, by comparing IS with IIS, we found apparent cell type abundance changes upon embryo implantation. In particular, the proliferating subsets of S1, S2, NK, M, DC and VEC were significantly increased. There were also massive gene expression changes in these cell types. Considering spatial relationships between cell types, uterine cell types other than PDZ are unable to directly communicate with embryonic cells during embryo implantation. Therefore, these changes were likely caused by PDZ indirectly via secreted signaling. Indeed, we found that many soluble factors from PDZ, such as Wnt4, Wnt5a and Wnt6, might have an influence on S1 and S2 cells. Wnt4 is most abundant in the decidual cells. In uterus-specific Wnt4 knockout mice, the embryos were able to attach to the uterine luminal epithelium, but they failed to successfully invade into the uterine stroma . During In conclusion, this study provided a comprehensive single-cell transcriptome atlas for mouse uterus at the invasion phase of embryo implantation. Our data present a valuable resource for deciphering the molecular mechanism underlying embryo implantation.Additional file 1: Table S1. The complete list of novel and known marker genes for each cell type.Additional file 2: Table S2. The complete list of signature genes for stromal cell clusters.Additional file 3: Table S3. The complete list of differentially expressed genes in IS compared to IIS for all cell types (logFC\u2009>\u20090.25 and P\u2009<\u20090.05)."} +{"text": "Cadia purpurea roots were investigated herein for the first time. The phytochemical study led to the isolation of two compounds, di-(2-methylheptyl) phthalate (1) and 13-O-pyrrolecarboxyl lupanine (2), from methanol roots extract of C. purpurea. The antibacterial activity results revealed that the n-hexane extract presented a better inhibitory value (13.8\u2009\u00b1\u20090.0\u2009mm) followed by chloroform (11.1\u2009\u00b1\u20090.4\u2009mm) and chloroform\u2009:\u2009methanol (1\u2009:\u20091) (10.7\u2009\u00b1\u20090.1\u2009mm) extracts against E. coli at the maximum dose of 100\u2009mg/mL. While, methanolic and ethanolic extracts displayed a mild activity against same bacterium at same dose. The methicillin resistant S. aureus was found with almost total resistance to all extracts up to the 100\u2009mg/mL. The chloroform\u2009:\u2009methanol (1\u2009:\u20091), chloroform, and n-hexane extracts recorded inhibition zone values better than chloramphenicol (7.2\u2009\u00b1\u20090.6\u2009mm at 30\u2009\u03bcg dose) against P. aeruginosa. The alcoholic extracts also exhibited an activity better than chloramphenicol up to 25\u2009mg/mL against same bacterium. Compound 2 produced a comparable inhibition value (9.6\u2009\u00b1\u20090.0\u2009mm to 18.5\u2009\u00b1\u20090.0\u2009mm) to that of chloramphenicol (21.5\u2009\u00b1\u20090.3\u2009mm) against E. coli at doses up to 1.0\u2009mg/mL; whereas, compound 1 showed a slight activity (7.1\u2009\u00b1\u20090.1\u2009mm\u201310.3\u2009\u00b1\u20090.0\u2009mm). Both compounds were found generally inactive against S. aureus, while they provided an activity better than chloramphenicol (7.2\u2009\u00b1\u20090.6\u2009mm) against P. aeruginosa with inhibition zones ranging from 7.1\u2009\u00b1\u20090.0\u2009mm to 9.0\u2009\u00b1\u20090.1\u2009mm for compound 1 and 7.2\u2009\u00b1\u20090.0\u2009mm to 10.6\u2009\u00b1\u20090.0\u2009mm for compound 2. Ethanolic and methanolic extracts exhibited a better DPPH radical scavenging activity and strong ferric ion reducing power at a concentration of 500\u2009\u03bcg/mL compared to the other extracts. Compound 1 also possessed a better anti-DPPH trapping activity than compound 2 . The compounds, however, indicated a weak ferric ion reduction power even at higher amount. In general, the observed antibacterial and antioxidant activities of isolated compounds and extracts were found to be dose-dependent. Conducting further biochemical investigations on all parts of this plant could provide opportunities of finding extra alkaloidal compounds and other phthalate derivatives with better biological activity.Phytochemicals and antibacterial and antioxidant activities of Cadia purpurea (Fabaceae) is a shrub which inhabits rift valley escarpment and bush land, commonly in altitudes about 1300\u20132700\u2009m. In Ethiopia. it is found in Tigray, Welo, Shewa, Hararge, and Bale regions. C. purpurea is also distributed in Eritrea, Yemen, Oman, North Somalia, and North Kenya + at m/z of 390 was showed which is compatible with the molecular composition of C24H38O4, 149.0 (100%), 167 (30.31%), 57.1 (16.65%), 71.1 (11.57%), and 279.1 (10.71%).Compound 1H NMR spectrum, a methylene proton signal at \u03b4H 4.22\u20134.24 , integrated to four protons, was observed. Besides, proton signals at \u03b4H 0.96\u20130.98 and 0.91\u20130.94 integrated each to six protons were shown implying that there were four methyl groups, in which the first two (C-8\u2032/8\u2033-CH3) were attached to sp3 methine carbons (C-2\u2032/2\u2033), while the latter two (C-7\u2032/7\u2033-CH3) were found to be terminals affixed to sp3 methylene carbons (C-6\u2032/6\u2033). The presence of two symmetric sp3 methine groups was also confirmed by the proton signal at \u03b4H 1.58\u20131.71 in the 1H NMR spectrum. Other eight symmetric methylene protons were appeared at \u03b4H 1.30\u20131.36 as multiplet integrated to a total of sixteen protons. Furthermore, two proton signals (integrated to two protons each) in the low field region at \u03b4H 7.72\u20137.74 and 7.62\u20137.64 were indicative of four symmetric aryl protons having the same coupling pattern. From 13C and DEPT-135 NMR spectra, twelve intense carbon peaks were clearly shown, which ascribed to twenty-four carbon numbers indicating that there is a symmetry in compound 1. For instance, the presence of a quaternary carbon signal at \u03b4 167.9 (C-7/8) and methylene carbon signal at \u03b4 67.7 (C-1\u2032/1\u2033) confirmed the occurrence of two symmetric aryl ester carbonyl groups. The 13C and DEPT-135 spectra also further supported this by showing eight symmetric carbons, i.e., six aromatic carbons at \u03b4 132.2 , 128.5 (C-2/5), and 131.0 (C-3/4). In addition, the occurrence of two carbon signals in the far right side of the 13C spectrum at \u03b4 10.1 and 13.1 indicated that there were four equivalence methyl groups, C-8\u2032/8\u2033-CH3 (attached to C-2/'2\u2033 at \u03b4 38.8), and terminal C-7\u2032/7\u2033-CH3 (affixed to C-6\u2032/6\u2033 at \u03b4 22.7). Moreover, other four symmetric methylene carbon peaks were observed at \u03b4 23.6 (C-3\u2032/3\u2033), 28.7 (C-4\u2032/4\u2033), 30.2 (C-5\u2032/5\u2033), and 22.7 (C-6\u2032/6\u2033). In the resulted IR spectrum, the absence of a sharp intense hydroxyl peak at its absorption region and the presence of a peak at the carbonyl functional group region (1727\u2009cm\u22121) along with the absorption bands of 1282 and 745\u2009cm\u22121, which were due to the ester C=O, C-O and -CH stretches of aromatic protons, respectively, directed that compound 1 exhibits a diester of phthalic acid. Based on the above discussed spectral data (1 was deduced as di-(2-methylheptyl) phthalate (1. Eventhough di-(2-methylheptyl) phthalate is a member of the phthalate groups, unlike to other phthalate esters, it is not a petrochemical and plasticizer. Most of the phthalates and their derivatives are petrochemicals which have been used as plasticizers in chemical industry to improve the plasticity and flexibility of the industrial products + was observed, 246.2 (100%), 134.05 (29.86%), 112.0 (weak), and 94.0 (weak).Compound 51\u2013154o along with a methylene carbon signal at the aliphatic region in the 13C spectrum. Besides, the relatively deshielded multiplet and triplet proton signals indicate the presence of methine groups attached to heteroatoms , as also reported elsewhere [\u03b4H 1\u20133\u2009ppm). From the 13C and DEPT-135 spectra, fifteen clear carbon signals were shown which represent a quaternary carbonyl carbon of the amide group at \u03b4 172.8 (C-2), nine sp3 methylene carbons at \u03b4 32.3 (C-3), 18.9 (C-4), 25.6 (C-5), 26.7 (C-8), 46.6 (C-10), 35.4 (C-12), 49.4 (C-15), 27.7 (C-14), and 51.2 (C-17), and five sp3 methine carbons appeared at \u03b4 60.7 (C-6), 33.9 (C-7), 32.2 (C-9), 57.9 (C-11), and 67.7 (C-13). Generally, the abovementioned proton and carbon signals were complementary and characteristics of lupanine skeleton. Besides, the absorption bands shown at 1616 and 2918\u20132855\u2009cm\u22121 in the IR spectrum, which belong to the amide carbonyl group (lactam C=O) and -CH stretch of the trans-quinolizidine, respectively, highlighted that compound 2 exhibits a lupanine skeleton. As also reported elsewhere [2 which was assured by three doublets of doublet proton signals occurred at \u03b4H 7.01 , 6.21\u20136.23 , and 6.90\u20136.92 assigned to the three olefinic methine protons of the pyrrolecarboxyl unit. Their corresponding carbon peaks appeared at \u03b4 123.5 (C-4\u2032), 109.4 (C-5\u2032) and 115.6 (C-6\u2032). In the DEPT-135 spectrum, the two quaternary carbon signals shown at \u03b4 122.2 and 161.7 belong to C-2\u2032 and the carbonyl carbon (C-1\u2032), respectively, of the substituent. Also, the carbonyl (C=O) and secondary -NH groups of the esterified pyrrolecarboxyl moiety showed absorption bands in the wavenumbers (cm\u22121) of 1692 and 3342 (broad), respectively, in the IR spectrum. This esterified pyrrolecarboxyl unit was incorporated to the lupanine skeleton of compound 2 at C-13 which was identified by the relatively deshielded carbon signal at \u03b4 67.7 (C-13) and triplet proton signal of H-13 at \u03b4H 5.19\u20135.21 . The detailed spectral information obtained of compound 2 is given in 2 was characterized as 13-O-pyrrolecarboxyl lupanine extracts were found effective against E. coli (>7\u2009mm inhibition zone) at all tested concentrations with the maximum inhibition diameters of 13.8\u2009\u00b1\u20090.0\u2009mm, 11.1\u2009\u00b1\u20090.0\u2009mm, and 10.7\u2009\u00b1\u20090.1\u2009mm, respectively, recorded at the maximal concentration of 100\u2009mg/mL, which were slightly comparable to chloramphenicol (24.5\u2009\u00b1\u20090.3\u2009mm at 30\u2009\u03bcg dose). The remaining lower concentrations, 50\u2009mg/mL, 25\u2009mg/mL, and 12.5\u2009mg/mL, of these extracts also produced an activity with corresponding inhibition zone values of 10.2\u2009\u00b1\u20090.4\u2009mm, 9.3\u2009\u00b1\u20090.0\u2009mm, and 8.7\u2009\u00b1\u20090.0\u2009mm recorded by n-hexane extract, 9.6\u2009\u00b1\u20090.3\u2009mm, 8.3\u2009\u00b1\u20090.0\u2009mm, and 7.2\u2009\u00b1\u20090.0\u2009mm recorded by chloroform extract, and 9.0\u2009\u00b1\u20090.3\u2009mm, 7.2\u2009\u00b1\u20090.0\u2009mm, and 7.0\u2009\u00b1\u20090.0\u2009mm recorded by chloroform/methanol (1\u2009:\u20091) extract. Interestingly, both the alcoholic extracts experienced a lesser activity against E. coli at concentrations of 100\u2009mg/mL and 50\u2009mg/mL with respective inhibitory values of 9.6\u2009\u00b1\u20090.1\u2009mm and 8.3\u2009\u00b1\u20090.1\u2009mm for the methanol extract and 8.0\u2009\u00b1\u20090.0\u2009mm and 7.6\u2009\u00b1\u20090.0\u2009mm for the ethanol extract, while no inhibited area was visible at the remaining concentrations of the extracts. The Gram-positive S. aureus strain was found to be susceptible only to the n-hexane and methanol extracts up to the 25\u2009mg/mL dose with measured inhibited area ranging from 7.6\u2009\u00b1\u20090.0\u2009mm to 10.2\u2009\u00b1\u20090.5\u2009mm and 7.3\u2009\u00b1\u20090.0\u2009mm to 8.7\u2009\u00b1\u20090.0\u2009mm, respectively. Whereas, completely no cleared zone was observed at the minimal concentration (12.5\u2009mg/mL) of these extracts. Also, all tested concentrations of chloroform, chloroform/methanol (1\u2009:\u20091), and ethanol extracts showed a totally zero zone of inhibition against S. aureus. On the other hand, the chloroform/methanol (1\u2009:\u20091) extract exerted a positive action on the growth of P. aeruginosa bacterium at all tested dilutions with measured cleared area of 10.0\u2009\u00b1\u20090.1\u2009mm at 100\u2009mg/mL, 9.7\u2009\u00b1\u20090.1\u2009mm at 50\u2009mg/mL, 8.4\u2009\u00b1\u20090.2\u2009mm at 25\u2009mg/mL, and 8.0\u2009\u00b1\u20090.0\u2009mm at 12.5\u2009mg/mL. The chloroform and n-hexane extracts also made an attempt to inhibit some area of P. aeruginosa growth at all concentrations by providing measured values of 7.7\u2009\u00b1\u20090.0 mm\u20139.8\u2009\u00b1\u20090.1\u2009mm and 7.3\u2009\u00b1\u20090.0 mm\u20138.9\u2009\u00b1\u20090.2\u2009mm, respectively. However, the methanolic and ethanolic extracts scored a slightly lower inhibitory values against P. aeruginosa at the doses up to 25\u2009mg/mL, but a nil inhibition zone at the least concentration (12.5\u2009mg/mL). To summarize, a better activity against E. coli, especially at the higher concentrations (100\u2009mg/mL and 50\u2009mg/mL), was noted in the nonalcoholic extracts though it looked like less as compared to chloramphenicol (24.5\u2009\u00b1\u20090.3\u2009mm at dose of 30\u2009\u03bcg). Whereas, the inhibitory action indicated by almost all extracts against S. aureus at all concentrations was noted weak in reference to chloramphenicol (18.8\u2009\u00b1\u20090.4\u2009mm). To the contrary, the inhibition zone values, scored by the extracts against P. aeruginosa, were found even better than that of chloramphenicol (7.2\u2009\u00b1\u20090.6\u2009mm).2 presented better inhibitory activity than compound 1 against E. coli with greater zone of inhibition of 18.5\u2009\u00b1\u20090.0\u2009mm recorded at the higher concentration of 1.0\u2009mg/mL, which was comparable to chloramphenicol (21.5\u2009\u00b1\u20090.3\u2009mm at dose of 30\u2009\u03bcg). The remaining concentrations, 0.5\u2009mg/mL, 0.3\u2009mg/mL, 0.1\u2009mg/mL, and 0.05\u2009mg/mL, of compound 2 also produced an activity against same bacterium with respective mean inhibition values of 16.2\u2009\u00b1\u20090.0\u2009mm, 13.8\u2009\u00b1\u20090.0\u2009mm, 10.0\u2009\u00b1\u20090.0\u2009mm, and 9.6\u2009\u00b1\u20090.0\u2009mm. Compound 1 displayed a moderate activity against E. coli at all concentrations (>7\u2009mm) with inhibition zones laid in the range of 7.2\u2009\u00b1\u20090.1 mm\u201310.3\u2009\u00b1\u20090.0\u2009mm. Whereas, against S. aureus, compound 1 showed a minor inhibitory effect only at concentrations of 1.0\u2009mg/mL (9.1\u2009\u00b1\u20090.1\u2009mm) and 0.5\u2009mg/mL (8.4\u2009\u00b1\u20090.0\u2009mm), but a zero effect at the remaining concentrations. Compound 2 was found totally inactive against S. aureus at all concentrations with nil inhibition zone values. However, the two compounds possessed an activity stronger than the chloramphenicol (7.2\u2009\u00b1\u20090.6\u2009mm) against P. aeruginosa bacterium almost at all dilutions, that is, compound 2 produced mean inhibition zone values ranging from 7.2\u2009\u00b1\u20090.6\u2009mm to 10.6\u2009\u00b1\u20090.0\u2009mm and that of compound 1 was found between 7.1\u2009\u00b1\u20090.0\u2009mm and 9.0\u2009\u00b1\u20090.1\u2009mm. 2 provided a notable activity against E. coli, especially at the higher concentration, which resulted in an inhibition value close to that of chloramphenicol; whereas, compound 1 was found with lesser activity as compared with compound 2 and chloramphenicol. However, against the methicillin resistant S. aureus strain, both compounds exhibited a negligible activity up to 1.0\u2009mg/mL in comparison to the chloramphenicol antibiotic (18.8\u2009\u00b1\u20090.4\u2009mm at the dose of 30\u2009\u03bcg). But, surprisingly, chloramphenicol had faced strong and unprecedented resistant from P. aeruginosa bacterium which led to even a smaller inhibition zone value (7.2\u2009\u00b1\u20090.6\u2009mm) than that of the compounds. Overall, the resulted inhibitory potential of the extracts and isolated compounds examined herein was found slightly comparable against E. coli, very weak against S. aureus, and stronger against P. aeruginosa in reference to the standard antibiotic drug, chloramphenicol (at 30\u2009\u03bcg dose). In fact, it is difficult to exactly compare the antimicrobial potency of extracts and/or isolates directly with synthesized standard antibiotics like chloramphenicol for similar doses, since there are huge differences between them in terms of chemical polarity, structural patterns, and others.As given in 1 and 2 of Cadia purpurea roots was evaluated against DPPH free radical and ferric ion oxidation. Six various dilutions of each extract and compound, including ascorbic acid, were prepared from their respective stock solutions (1\u2009mg/mL). Each solution was subjected to the UV-Vis spectrophotometer to measure their absorbance against DPPH radical and ferric reducing power at 517 and 700\u2009nm, respectively. Resulted values of duplicates were expressed as mean\u2009\u00b1\u2009standard deviation.In vitro antioxidant potential of four extracts and two isolated compounds DPPH free radical scavenging activity: the DPPH free radical scavenging potential of extracts and isolated compounds was determined compared to ascorbic acid (AA). The obtained result showed that activity of the extracts and compounds against DPPH radical was dose-dependent, in which the DPPH scavenging activity percentage was directly correlated with concentrations .50 values of 12.9 and 16.03\u2009\u03bcg/mL at 500\u2009\u03bcg/mL concentration. The chloroform and chloroform/methanol (1\u2009:\u20091) extracts provided the comparatively lesser anti-DPPH activity percentage of 77.07\u2009\u00b1\u20090.00 and of 79.38\u2009\u00b1\u20090.16 (26.14\u2009\u03bcg/mL of IC50) at same concentration. In reference to the observed anti-DPPH potency of ascorbic acid at 500\u2009\u03bcg/mL, however, the inhibitory activity against the free radical of the tested extracts was found to be weak.As given in 1 presented a greater anti-DPPH activity percentage of 82.69\u2009\u00b1\u20090.11 at concentration of 500\u2009\u03bcg/mL with an IC50 value of 7.99\u2009\u03bcg/mL. The quinolizidine alkaloid compound 2 exhibited the smaller scavenging percentage value for same concentration.Of the compounds evaluated, compound 4\u2009\u03bcg/mL) . Here, t3+) ion into its ferrous (Fe2+) of extracts and isolates was known by first observing a change in color of the reaction solutions from yellow to green and then measuring the absorbance at 700\u2009nm [Ferric reducing antioxidant power examination: the potential of reducing ferric at same amount toward DNA gyraseB protein of E. coli selected from the Protein Data Bank (PDB ID: 6F86). The docking analysis result is observed in compound 2 which is comparable with that of ciprofloxacin . As shown in 2 makes three H-bond interactions with Thr-165 via water, Gly-77, and Ile-78 residual amino acids. Besides, this compound coordinates to Pro-79 including a Van der Walls interaction with Asp-73. The hydrophobic and \u03c0-anion binding ability of compound 1 with Ile-78 and Glu-50 binding pockets, respectively, is the only drug-likeness indicator of the compound in comparison to ciprofloxacin. Whereas, the ciprofloxacin-like property of compound 2 is recognized only via the formation of a H-bond with Thr-165 active binding site of 6F86.Isolated compounds s result revealed1) and 13-O-pyrrolecarboxyl lupanine (2), were isolated from methanolic extract of Cadia purpurea roots for the first time. In vitro antibacterial and antioxidant activities of isolated compounds and crude extracts were also evaluated, and the results showed that n-hexane, chloroform, and chloroform/methanol (1\u2009:\u20091) extracts were found effective against E. coli at all tested concentrations with the maximum inhibition diameters of 13.8\u2009\u00b1\u20090.0\u2009mm, 11.1\u2009\u00b1\u20090.0\u2009mm, and 10.7\u2009\u00b1\u20090.1\u2009mm, respectively, recorded at the maximal concentration of 100\u2009mg/mL, which were slightly comparable to chloramphenicol (24.5\u2009\u00b1\u20090.3\u2009mm at 30\u2009\u03bcg dose). The methanolic and ethanolic extracts, however, experienced a smaller activity against E. coli merely at concentrations of 100\u2009mg/mL and 50\u2009mg/mL with respective inhibitory values of 9.6\u2009\u00b1\u20090.1\u2009mm and 8.3\u2009\u00b1\u20090.1\u2009mm, and 8.0\u2009\u00b1\u20090.0\u2009mm and 7.6\u2009\u00b1\u20090.0\u2009mm. The methicillin resistant S. aureus strain was found to be sensitive only to the n-hexane and methanol extracts up to the 25\u2009mg/mL dose with measured inhibited area ranging from 7.6\u2009\u00b1\u20090.0\u2009mm to 10.2\u2009\u00b1\u20090.5\u2009mm, and 7.3\u2009\u00b1\u20090.0\u2009mm to 8.7\u2009\u00b1\u20090.0\u2009mm, respectively. Whereas the chloroform, chloroform/methanol (1\u2009:\u20091), and ethanol extracts did not indicate any sign of activity against S. aureus even at the higher concentration of 100\u2009mg/mL. On the other hand, the chloroform/methanol (1\u2009:\u20091) extract exerted an inhibitory action on the growth of P. aeruginosa bacterium at all tested dilutions with measured clear area of 8.0\u2009\u00b1\u20090.0\u2009mm\u201310.0\u2009\u00b1\u20090.1\u2009mm. The chloroform and n-hexane extracts also attempted to inhibit some growth of P. aeruginosa at all concentrations with corresponding measured values of 7.7\u2009\u00b1\u20090.0\u2009mm\u20139.8\u2009\u00b1\u20090.1\u2009mm and 7.3\u2009\u00b1\u20090.0\u2009mm\u20138.9\u2009\u00b1\u20090.2\u2009mm. However, the methanolic and ethanolic extracts scored a slightly lower inhibitory values against P. aeruginosa up to 25\u2009mg/mL concentration. Compound 2 presented a better inhibitory effect against E. coli than compound 1 with a comparable inhibition value (18.5\u2009\u00b1\u20090.0\u2009mm) to chloramphenicol (21.5\u2009\u00b1\u20090.3\u2009mm at dose of 30\u2009\u03bcg) recorded at the higher concentration of 1.0\u2009mg/mL. This compound 2 also produced an activity on same bacterium at the concentrations of 0.5\u2009mg/mL, 0.3\u2009mg/mL, 0.1\u2009mg/mL, and 0.05\u2009mg/mL with zone of inhibition values ranging between 9.6\u2009\u00b1\u20090.0\u2009mm and 16.2\u2009\u00b1\u20090.0\u2009mm. Compound 1 displayed a moderate activity (>7\u2009mm) against E. coli at all concentrations with inhibition zones laid in the range of 7.2\u2009\u00b1\u20090.1\u2009mm\u201310.3\u2009\u00b1\u20090.0\u2009mm. Whereas, against S. aureus, compound 1 showed a minor inhibitory effect only at concentrations of 1.0\u2009mg/mL (9.1\u2009\u00b1\u20090.1\u2009mm) and 0.5\u2009mg/mL (8.4\u2009\u00b1\u20090.0\u2009mm) and compound 2 was found totally inactive at all concentrations. The two compounds, however, possessed an activity stronger than the chloramphenicol (7.2\u2009\u00b1\u20090.6\u2009mm) against P. aeruginosa bacterium almost at all dilutions with mean inhibition zone values ranging from 7.2\u2009\u00b1\u20090.6\u2009mm to 10.6\u2009\u00b1\u20090.0\u2009mm of compound 2 and 7.1\u2009\u00b1\u20090.0\u2009mm to 9.0\u2009\u00b1\u20090.1\u2009mm of compound 1. The obtained antibacterial result generally revealed that nonalcoholic extracts were noted with a better activity against E. coli, especially at the higher concentrations (100\u2009mg/mL and 50\u2009mg/mL), though they seemed like weak compared to the chloramphenicol (24.5\u2009\u00b1\u20090.3\u2009mm at dose of 30\u2009\u03bcg). Compound 2 also provided a notable activity against E. coli, especially at the higher concentration, which resulted in an inhibition value close to that of chloramphenicol (21.5\u2009\u00b1\u20090.3\u2009mm); whereas, compound 1 was found with weaker activity as compared with compound 2 and chloramphenicol. On the other hand, against the methicillin resistant S. aureus strain, both compounds and all the extracts exhibited a negligible activity up to the higher dosage in reference to the chloramphenicol antibiotic (18.8\u2009\u00b1\u20090.4\u2009mm). On the contrary, the inhibition zone values, scored by the extracts and compounds against P. aeruginosa, were found even better than that of chloramphenicol (7.2\u2009\u00b1\u20090.6\u2009mm). Regarding the antioxidant potency, ethanolic and methanolic extracts exhibited a better DPPH radical scavenging activity at concentration of 500\u2009\u03bcg/mL, whereas the chloroform and chloroform/methanol (1\u2009:\u20091) extracts were observed weak with IC50 values of 27.52 and 26.14\u2009\u03bcg/mL. Comparatively strong ferric ion reducing power was also shown in the alcoholic extracts with 0.810\u2009\u00b1\u20090.001 absorbance of methanol and 0.788\u2009\u00b1\u20090.000 of ethanol at the maximum amount (500\u2009\u03bcg/mL). Compound 1 indicated a promising trapping potential against DPPH radical , while compound 2 recorded a higher IC50 value (58.34\u2009\u03bcg/mL) at concentration of 500\u2009\u03bcg/mL. However, at same dosage, the isolated compounds indicated a weak power of ferric ion reduction with recorded absorbance of 0.761\u2009\u00b1\u20090.002 by compound 1 and 0.458\u2009\u00b1\u20090.001 of compound 2. The molecular docking result revealed that compound 1 shows two hydrophobic (with Ile-78 and Ile-94), a \u03c0-anion (with Glu-50) and Van der Walls interactions with some amino residues of E. coli gyraseB; whereas, compound 2 makes three H-bond interactions with Thr-165, Gly-77, and Ile-78 in addition to the Van der Walls interaction with Asp-73. In conclusion, the present findings indicated that the resulted antibacterial inhibitory potential of the extracts and isolated compounds was found slightly active against E. coli, very weak against S. aureus, and stronger against P. aeruginosa as compared to chloramphenicol (at 30\u2009\u03bcg dose); and the entire antibacterial and antioxidant activities of tested analytes were noted as dose-dependent. Besides, since other quinolizidine alkaloids (similar to compound 2) were also reported from the leaves of Ethiopian Cadia purpurea, conducting further biochemical investigations on all parts of the plant could provide opportunities of finding additional alkaloidal compounds and other phthalates with powerful biological activities.In this study, two compounds, di-(2-methylheptyl) phthalate ("} +{"text": "Paratylenchus spp., are relatively small nematodes that can feed on a wide range of host plants. The morphological identification of this nematode is greatly hampered by their small size and variable characters. This study provides the first report of Paratylenchus lepidus from Vietnam with a combination of morphological and molecular characterizations. The 28S rDNA phylogenetic tree of the genus and the first COI mtDNA barcode of this species are also provided.The pin nematodes, Paratylenchus is commonly known as pin nematodes that are ectoparasites and can be frequently found at high density in perennial plants, hop gardens, orchards, or forest trees Kuntze) in Vietnam. Nematodes were extracted using the modified Baermann tray method (COI mtDNA gene were amplified using D2A/D3B (5\u2032\u2013ACAAGTACCGTGGGGAAAGTTG\u20133\u2032/5\u2032\u2013TCGGAAGGAACCAGCTACTA\u20133\u2032) (www.geneious.com). The best fit model was chosen using Mega 7 and phylogenetic analysis was done following Nguyen et al. (2019c).Soil and root samples were collected from the rhizosphere of green tea (y method . After tACTA\u20133\u2032) and JB3/n\u2009=\u200920 (\u2640\u2640): L\u2009=\u2009340\u2009\u00b1\u200920 (307-371) \u00b5m, a\u2009=\u200925\u2009\u00b1\u20091 (22-27), b\u2009=\u20094.1\u2009\u00b1\u20090.3 (3.7-4.6), c\u2009=\u200911.5\u2009\u00b1\u20091.4 (9.8-13.8), c\u2032\u2009=\u20093.5\u2009\u00b1\u20090.4 (3.0-4.1), V%\u2009=\u200982\u2009\u00b1\u20091 (81-84), Lip height\u2009=\u20092.7\u2009\u00b1\u20090.5 (1.9-3.6) \u00b5m, Lip width\u2009=\u20094.9\u2009\u00b1\u20090.5 (4.2-5.9) \u00b5m, Stylet\u2009=\u200925\u2009\u00b1\u20091 (24-27) \u00b5m, Median bulb length\u2009=\u200915.2\u2009\u00b1\u20092 (12.8-18.0) \u00b5m, Median bulb width\u2009=\u20096.9\u2009\u00b1\u20090.6 (5.9-7.7) \u00b5m, SE pore\u2009=\u200975\u2009\u00b1\u20094 (67-81) \u00b5m, Pharynx\u2009=\u200984\u2009\u00b1\u20093 (79-91) \u00b5m, Body width\u2009=\u200913.8\u2009\u00b1\u20090.4 (13.3-14.5) \u00b5m, Vulval body diam.\u2009=\u200912.2\u2009\u00b1\u20090.4 (11.3-12.6) \u00b5m, Anal body diam.\u2009=\u20098.5\u2009\u00b1\u20090.4 (7.9-9.5) \u00b5m, Tail length\u2009=\u200930\u2009\u00b1\u20093 (26-35) \u00b5m.Paratylenchus lepidus is characterized by having a slender body, curved ventrally; lateral field with four incisures; lip region weakly sclerotized, continuous to body contour; median bulb elongate with a distinct valve; isthmus slender, surrounded by nerve ring; basal bulb pyriform; secretory-excretory pore located at level of basal bulb to pharyngo-intestinal junction; hemizonid located just anterior to secretory-excretory pore; gonad monodelphic, post uterine sac absent; vulval lips not protruding but having prominent advulval \ufb02ap; tail curved ventrally with a finely rounded to bluntly pointed terminus was 99.7% similar (2\u2009bp difference) to the sequence of P. lepidus from GenBank (accession number: MK886692). The phylogenetic tree based on 28S rDNA sequences showed that the sequence of P.\u00a0lepidus from Vietnam was placed together with the sequence of P. lepidus from GenBank (100% PP) (COI mtDNA sequence of P. lepidus from Vietnam (418\u2009bp long) was also obtained and submitted to GenBank under the accession number MT828831.The 28S rDNA sequence of 100% PP) . A COI mP. lepidus from green tea in Vietnam is in agreement with the description of the type population (P. lepidus in Vietnam. The first COI mtDNA sequence of P. lepidus is also provided to serve as a molecular barcode for molecular identification of Paratylenchus species in the future.Morphology of pulation with smapulation . In this"} +{"text": "Two copper(II) cations are situated in distorted square-pyramidal environment, while two copper(II) cations are located in a slightly square-planar geometry. One bridging acetate group acting in an \u03b71:\u03b71-\u03bc2-mode connects two copper(II) ions, while another bridging acetate group connects three copper(II) ions in an \u03b71:-\u03b72\u2013\u03bc3-mode.In the title Schiff base tetra\u00adnuclear copper(II) complex, two discrete environments are present in the structure: CuNO 3-acetato)(\u03bc2-acetato)\u00adbis\u00adethyl\u00adidene]amino}\u00adpropan-2-olato)tetra\u00adcopper(II) monohydrate, [Cu4(C19H19N2O3)2(CH3CO2)2]\u00b7H2O, corresponds to a non-symmetric tetra\u00adnuclear copper complex. The complex exhibits one ligand mol\u00adecule that connects two copper CuII metal centres via its ethano\u00adlato oxygen anion acting in a \u03bc2-mode and one ligand mol\u00adecule that connects three copper CuII metal centres via its ethano\u00adlato oxygen anion acting in a \u03bc3-mode. One bridging acetate group acting in an \u03b71:\u03b71-\u03bc2-mode connects two copper(II) ions while another bridging acetate group connects three copper(II) ions in an \u03b71:-\u03b72-\u03bc3-mode. A chair-like Cu3O3 structure is generated in which the two CuO4N units are connected by one \u03bc2-O ethano\u00adlate oxygen atom. These two units are connected respectively to the CuO3N unit via one \u03bc3-O ethano\u00adlate oxygen atom and one \u03bc2-O atom from an acetate group. The \u03bc3-O atom also connects one of the CuO4N units and the CuO3N unit to another CuO3N unit, which is out of the chair-like structure. Each of the two penta\u00adcoordinated CuII cations has a distorted NO4 square-pyramidal environment. The geometry of each of the two CuNO3 units is best described as a slightly square-planar environment. A series of intra\u00admolecular O\u2014H\u22efO hydrogen bonds is observed. In the crystal, the units are connected by inter\u00admolecular C\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, thus forming sheets parallel to the ac planeThe title mol\u00adecular structure, namely, (\u03bc Mixing H3L and hydrated copper acetate yielded a tetra\u00adnuclear complex formulated as [Cu4L2(CH3CO2)2]\u00b7H2O in which the ligand acts in its tri-deprotonated \u22123L form. In the tetra\u00adnuclear complex, one of the \u22123L anions acts in \u03bc2-mode, connecting the two penta\u00adcoordinated CuII cations. The second \u22123L anion acts in \u03bc3 mode, connecting the two tetra\u00adcoordinated CuII cations and one of the penta\u00adcoordinated CuII cations. The second penta\u00adcoordinated CuII cation is connected to the two tetra\u00adcoordinated CuII cations via an acetate group acting in \u03b71:\u03b72-\u03bc3 mode. Additionally, the two penta\u00adcoordinated CuII cations are connected by an acetate group acting in \u03b71:\u03b71-\u03bc2 mode. For each ligand, the azomethine nitro\u00adgen atom and the phenolate oxygen atom of one arm are both linked to one CuII cation while the corresponding atoms of the other arm are bonded to another CuII cation. No phenolate oxygen atom acts in bridging mode. In one ligand the ethano\u00adlate oxygen atom bridges the two penta\u00adcoordinated CuII cations, and in the second ligand the ethano\u00adlate oxygen atom bridges the two tetra\u00adcoordinated CuII cations and one penta\u00adcoordinated CuII cation. The two \u22123L ligands are coordinated differently in hexa\u00addentate and hepta\u00addentate fashions. Four five-membered CuOCCN rings and four six-membered CuOCCCN rings are formed upon the coordination of the ligand mol\u00adecules. In the tetra\u00adnuclear complex, two discrete CuO4N and CuO3N units are observed.\u03c4 parameter around the metal ion are \u03c4 = 0.1103 for Cu1 and \u03c4 = 0.1887 for Cu2. For Cu1 and Cu2, the basal planes are occupied by one phenolate oxygen anion, one azomethine nitro\u00adgen atom, one ethano\u00adlate oxygen atom and one oxygen atom from the \u03b71:\u03b71-\u03bc2 acetate group, the apical position being occupied by an ethano\u00adlate oxygen atom from a second ligand mol\u00adecule for Cu1 and an oxygen atom from the \u03b71:\u03b72-\u03bc3 acetate group for Cu2. The atoms forming the basal plane for Cu1 are almost coplanar (r.m.s. deviation = 0.1088\u2005\u00c5) and the Cu1 atom is displaced toward the O5 atom, which occupies the apical position, by 0.0545\u2005(2)\u2005\u00c5. The Cu1\u2014O5 distance of 2.749\u2005(3)\u2005\u00c5 is longer than the distances between Cu1 and the atoms in the basal plane , as expected for a Jahn\u2013Teller distortion \u201395.10\u2005(14)\u00b0 and 169.71\u2005(16)\u2013176.33\u2005(14)\u00b0, respectively. The atoms forming the basal plane for Cu2 are less coplanar than those around Cu1 (r.m.s. deviation = 0.2086\u2005\u00c5) and the Cu2 atom is displaced toward the O8 atom, which occupies the apical position, by 0.0808\u2005(1)\u2005\u00c5. The from Cu2\u2014O8 distance of 2.703\u2005(4)\u2005\u00c5 is longer than those to atoms in the equatorial plane . As observed for Cu1, Jahn\u2013Teller distortion \u201396.89\u2005(14)\u00b0 and 161.66\u2005(15)\u2013173.00\u2005(15)\u00b0, respectively. The bond lengths involving the \u03bc2-bridging ethano\u00adlato oxygen atom and the copper cations are asymmetrical: Cu1\u2014O2 = 1.916\u2005(3)\u2005\u00c5 and Cu2\u2014O2 = 1.920\u2005(3)\u2005\u00c5. The distances between the \u03bc3-bridging ethano\u00adlato oxygen atom and the copper cations are very different: Cu1\u2014O5 = 2.749\u2005(3)\u2005\u00c5, Cu3\u2014O5 = 1.907\u2005(3)\u2005\u00c5 and Cu4\u2014O5 = 1.921\u2005(3)\u2005\u00c5. The copper cations Cu3 and Cu4 are coordinated by one ethano\u00adlato oxygen anion, one phenoxo oxygen anion, one azomethine nitro\u00adgen atom of the ligand and one oxygen atom of a \u03b71:\u03b72-\u03bc3 acetate group (O8 for Cu3 and O7 for Cu4). The Cu3\u2014O4 [1.873\u2005(3)\u2005\u00c5], Cu3\u2014O5 [1.907\u2005(3)\u2005\u00c5], Cu3\u2014N3 [1.947\u2005(4)\u2005\u00c5], Cu3\u2014O8 [1.957\u2005(3)\u2005\u00c5], Cu4\u2014O6 [1.869\u2005(3)\u2005\u00c5], Cu4\u2014O5 [1.921\u2005(3)\u2005\u00c5], Cu4\u2014N4 [1.962\u2005(4)\u2005\u00c5] and Cu4\u2014O7 [1.955\u2005(3)\u2005\u00c5] distances are in close proximity to values reported for copper(II) complexes with analogous Schiff base ligands and 0.1801 (Cu4) suggested distorted square-planar geometries. For the two copper cations the cisoid angles are in the ranges 86.17\u2005(14)\u201393.29\u2005(15)\u00b0 for Cu3 and 84.04\u2005(14)\u201396.93\u2005(14)\u00b0 for Cu4 and the transoid angles are O4\u2014Cu3\u2014O5 = 177.07\u2005(15)\u00b0, O8\u2014Cu3\u2014N3 = 173.28\u2005(15)\u00b0, O6\u2014Cu4\u2014O5 = 170.48\u2005(14)\u00b0 and O7\u2014Cu3\u2014N4 = 164.11\u2005(15)\u00b0. The C\u2014N bonds are in the range 1.291\u2005(6)\u20131.300\u2005(6)\u2005\u00c5, indicative of double-bond character and the presence of the imino groups in the two ligands.Atoms Cu1 and Cu2 are penta\u00adcoordinated and their environments can be best described as slightly distorted square-pyramidal. The Addison phenoxo are observed ethyl\u00adidene)]-2-hy\u00addroxy\u00adpro\u00adpane-1,3-di\u00adamine is widely used in coordination chemistry. The current release of the CSD ethyl\u00adidene)]-2-hy\u00addroxy\u00adpropane-1,3-di\u00adamine (HL3) was prepared from 1-(2-hy\u00addroxy\u00adphen\u00adyl)ethanone and 2-hy\u00addroxy\u00adpropane-1,3-di\u00adamine in a 2:1 ratio in ethanol according to a slight modification of a literature method : 3538 (OH), 3268 (OH), 1605 (C=N), 1538 (C=C), 1528 (C=C), 1455 (C=C), 1247 (C\u2014O), 1043, 760. Analysis calculated for C19H22N2O3: C, 69.92; H, 6.79; N, 8.58. Found: C, 69.90; H, 6.76; N, 8.56%.The ligand 3CO2)2\u00b7(H2O) in 5\u2005mL of ethanol was added to a solution of H3L in 10\u2005mL of ethanol at room temperature. The initial yellow solution immediately turned deep green and was stirred for 30\u2005min before being filtered. The filtrate was kept at 298\u2005K. After one week, light-green crystals suitable for X-ray diffraction were collected and formulated as [Cu4L2(CH3CO2)2]\u00b7H2O. FT\u2013IR : 3404, 1601, 1532, 1332, 1299, 895, 760. Analysis calculated for C42H46Cu4N4O11: C, 48.64; H, 4.47; N, 5.40. Found: C, 48.60; H, 4.49; N, 5.44%.A solution of Cu were geometrically optimized and refined using a riding model (AFIX instructions) with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C) for CH3 and OH groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022002225/ex2053sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022002225/ex2053Isup3.hklStructure factors: contains datablock(s) I. DOI: 2154581CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A precise crystal-structure analysis using a neutron diffractometer with high-power neutron sources at the J-PARC facility has been performed on non-deuterated triglycine sulfate at 20\u2005K and 298\u2005K and a new double-potential-well disorder model for the O\u2014H\u22efO hydrogen bond in the 298\u2005K structure is proposed. 2H6NO2+\u00b7SO42\u2212\u00b7C2H5NO2 (or C6H17N3O10S), non-deuterated triglycine sulfate (HTGS) at 20\u2005K and 298\u2005K were undertaken using time-of-flight neutron diffraction data. At 20\u2005K for the O\u2014H\u22efO hydrogen bond between the glycinium cation and the zwitterionic, unprotonated glycine mol\u00adecule that is associated with the ferroelectric behaviour of HTGS, O\u2014H = 1.070\u2005(3), H\u22efO = 1.408\u2005(3) [\u03b4 = 0.338\u2005(4)], O\u22efO = 2.4777\u2005(15)\u2005\u00c5 and O\u2014H\u22efO = 179.0\u2005(4)\u00b0, which is in good agreement with previous studies. Two reasonable structures for the same three atoms were refined for the 298\u2005K dataset. One is a single-minimum potential-energy model, with O\u2014H = 1.090\u2005(12), H\u22efO = 1.361\u2005(12) [\u03b4 = 0.271\u2005(17)], O\u22efO = 2.450\u2005(7)\u2005\u00c5 and O\u2014H\u22efO = 179.2\u2005(10)\u00b0, having the H atom with a large ellipticity along the bond path between the O atoms. The other is a double-minimum potential-energy model having two H atom sites with occupancies of 0.876\u2005(8) and 0.124\u2005(8): for the major occupancy component, O\u2014H = 1.065\u2005(12), H\u22efO = 1.387\u2005(12), O\u22efO = 2.451\u2005(7)\u2005\u00c5 and O\u2014H\u22efO = 178.2\u2005(11)\u00b0 and for the minor component, O\u2014H = 1.06\u2005(4), H\u22efO = 1.41\u2005(4), O\u22efO = 2.451\u2005(7)\u2005\u00c5 and O\u2014H\u22efO = 166\u2005(2)\u00b0. These models did not show any significant differences in R factors. In addition, the unit-cell parameters and other structural parameters of HTGS did not show any major differences compared to those of partially deuterated TGS and fully deuterated TGS for both 20\u2005K and 298\u2005K.Precise single-crystal structure analyses of the title compound, bis\u00ad(glycinium) sulfate\u2013glycine (1/1), 2C The authors also proposed a hydrogen-bonding scheme and pointed out that the hydrogen atom that lies between the oxygen atom of the carboxyl group in the glycine III cation (GIII) and the O atom in the glycine II mol\u00adecule (GII) plays a crucial role in the dipole reversal. Many structural studies on TGS have subsequently been conducted (see Database survey): most of them were X-ray diffraction studies, but some of them were neutron diffraction studies. The atomic coordinates of non-deuterated TGS , including those of the hydrogen atoms at room temperature, were first revealed using single-crystal neutron diffraction group in each glycine mol\u00adecule and those in sulfuric acid mol\u00adecules at 40\u2005K and 180\u2005K , where deuterium replaced the H atoms except for the hydrogen atoms of the methyl\u00adene , all the hydrogen atoms in the glycine mol\u00adecules and sulfuric acid mol\u00adecules are substituted by deuterium atoms: the crystal structures did not show major changes between 20\u2005K and 295\u2005K exist as monoprotonated C2H6NO2+ glycinium ions. The most significant feature of these glycine/glycinium species are the N\u2014C\u2014C\u2014O(H) torsion angles \u00b0 for N11\u2014C17\u2014C19\u2014O7, \u22121.5\u2005(1)\u00b0 for N14\u2014C15\u2014C18\u2014O10 and \u22121.4\u2005(1)\u00b0 for N21\u2014C20\u2014C16\u2014O2. The sulfate ion shows its expected tetra\u00adhedral shape with bond distances of 1.480\u2005(2)\u2005\u00c5 (S1\u2014O4), 1.470\u2005(2)\u2005\u00c5 (S1\u2014O5), 1.477\u2005(2)\u2005\u00c5 (S1\u2014O6) and 1.472\u2005(2)\u2005\u00c5 (S1\u2014O8) and bond angles of 110.3\u2005(1)\u00b0 (O4\u2014S1\u2014O5), 107.9\u2005(1)\u00b0 (O4\u2014S1\u2014O6), 108.7\u2005(1)\u00b0 (O4\u2014S1\u2014O8), 109.7\u2005(1)\u00b0 (O5\u2014S1\u2014O6), 110.6\u2005(1)\u00b0 (O5\u2014S1\u2014O8) and 109.7\u2005(1)\u00b0 (O6\u2014S1\u2014O8). The slight differences among these distances and angles may arise because of the different hydrogen bonds accepted by these O atoms. Numerous N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds (see supporting information) are formed between the glycine or glycinium species and the sulfate ions; four N\u2014H\u22efO hydrogen bonds and one O\u2014H\u22efO hydrogen bond are formed by GI, five N\u2014H\u22efO hydrogen bonds are formed by GII and five N\u2014H\u22efO hydrogen bonds with the sulfate ion and one O\u2014H\u22efO hydrogen bond to the glycine mol\u00adecule is formed by GIII.The refined structures at 20\u2005K are shown in Figs. 1i bond lengths for HTGS, DTGS and FDTGS at low temperature are listed in Table\u00a01et al. 1997The lattice constants and the key O15\u2014H15\u22efO3B amino group [refined site occupancies = 0.874\u2005(8):0.126\u2005(8)] in the GI cation and the O\u2014H\u22efO association for GIII and GII. Two models were refined considering the H atom between O15 in GIII and O3 in GII. For one model (298\u2005K model 1), the H15 atom was refined with a large ellipticity along the bond path between O15 and O3 as a single minimum potential energy structure [Fig.\u00a05a)]. A double-minimum potential-energy structure could be deduced because the distance between O15 and O3i did not increase with an increase in the temperature; thus for the other model (298\u2005K model 2), a pair of hydrogen atoms were refined along the bond path between O15 and O3i, the double-minimum potential structure [Fig.\u00a05b)].The refined structures at 298\u2005K are shown in Figs. 3i hydrogen bond at 298\u2005K are summarized in Table\u00a02i in GII. On the other hand, the distance between O15 and H15 [1.090\u2005(12)\u2005\u00c5] is almost the same as that at 20\u2005K despite there being no distance restraint for the H15\u22efO3i separation. Therefore, the mixed structure (model 2) of the major ferroelectric phase and minor paraelectric phase is strongly suggested, because the occupancies of N11 and N11B and H15 and H3i are related by symmetry.The key parameters for the O15\u2014H15\u22efO3et al., 1997The unit-cell parameters and bond lengths for HTGS, DTGS, and FDTGS at 298\u2005K are listed in Table\u00a02TC for FDTGS increased by approximately 12\u2005K compared to HTGS. Choudhury & Chitra (2008B (12%); this occupancy ratio is in excellent agreement with the results in this study. For the hydrogen atom between the oxygen atom of the carboxyl group in GIII and that in the GII, the O\u22efO distance was 2.470\u2005(9)\u2005\u00c5, and the H atom was approximately 0.241\u2005\u00c5 closer to the GIII O atom than that in GII. They concluded that the structure of HTGS at room temperature has a single minimum potential energy in the O\u2014H\u22efO hydrogen-bond path between GIII and GII. In this study, two reasonable structures were refined as a single-minimum potential-energy model and a double-minimum model without any significant differences. Therefore, we conclude that there is a significant possibility of a double-minimum potential-energy model for HTGS at 298\u2005K.In the previous studies using single-crystal neutron diffraction, Kay & Kleinberg 1973 proposeditra 2008 proposedsupporting information) and no additional inter\u00admolecular inter\u00adactions were found. Therefore, the 20\u2005K and 298\u2005K structures form essentially the structural motif of a three-dimensional network of N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds between glycinium cations, glycine mol\u00adecules and sulfate ions.Hydrogen bonds in the refined structures were consistent with those reported previously ; TGLYSU15 and sulfuric acid was added to 50\u2005ml of water in a 100\u2005ml beaker. They were dissolved by heating at \u223c313\u2005K with a 300 r.p.m. magnetic stirrer. After completely dissolving them, plastic films were double-wrapped around the beaker, and some holes were knocked in the films to evaporate the water slowly. The beaker was left to stand at \u223c293\u2005K. HTGS was crystallized after approximately a month, and then the solution was filtered. The collected crystals were dried in a desiccator at \u223c293\u2005K.et al., 2016x, y, \u03bb) were measured in 16 different orientations for each dataset. The measurement time was 1.5\u2005h for one orientation; the raw data were processed using STARGazer slice maps and merged TOF profiles.Crystal data, data collection, and structural refinement details are summarized in Table\u00a03SHELXL2018 between O15 and O3 to minimize the model dependence. A nuclear density distribution global, 20K, 298KModel1, 298KModel2. DOI: 10.1107/S2056989022000858/hb800420Ksup2.hklStructure factors: contains datablock(s) 20K. DOI: Click here for additional data file.10.1107/S2056989022000858/hb800420Ksup5.cmlSupporting information file. DOI: 10.1107/S2056989022000858/hb8004298KModel1sup3.hklStructure factors: contains datablock(s) 298KModel1. DOI: Click here for additional data file.10.1107/S2056989022000858/hb8004298KModel1sup6.cmlSupporting information file. DOI: 10.1107/S2056989022000858/hb8004298KModel2sup4.hklStructure factors: contains datablock(s) 298KModel2. DOI: Click here for additional data file.10.1107/S2056989022000858/hb8004298KModel2sup7.cmlSupporting information file. DOI: 2144164, 2144163, 2144162CCDC references: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The packing of the title compound features C\u2014H\u22efO hydrogen bonds, C\u2014F\u22ef\u03c0 inter\u00adactions, aromatic \u03c0\u2013\u03c0 stacking and short Br\u22efO contacts. 14H8Br2FN3O2, the nitro-substituted benzene ring and the 4-fluoro\u00adphenyl ring form a dihedral angle of 65.73\u2005(7)\u00b0. In the crystal, mol\u00adecules are linked into chains by C\u2014H\u22efO hydrogen bonds running parallel to the c-axis direction. The crystal packing is consolidated by C\u2014F\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions, and short Br\u22efO [2.9828\u2005(13)\u2005\u00c5] contacts are observed. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions to the crystal packing are from H\u22efH (17.4%), O\u22efH/H\u22efO (16.3%), Br\u22efH/H\u22efBr (15.5%), Br\u22efC/C\u22efBr (10.1%) and F\u22efH/H\u22efF (8.1%) contacts.In the title compound, C R\u2014N=N\u2014R\u2032, where R and R\u2032 can be either aryl, hetrocycle or alkyl functional groups. They find many applications such as mol\u00adecular switches, optical data storage, anti\u00admicrobial agent, colour-changing materials, non-linear optics, mol\u00adecular recognition, dye-sensitized solar cells, liquid crystals, catalysis, etc. \u00b0. All of the other bond lengths and angles in the title compound are similar to those in the related azo compounds reported in the Database survey.As shown in Fig.\u00a01Cg1 = 3.4095\u2005(12)\u2005\u00c5; C\u2014F\u22efCg1 = 136.95\u2005(9)\u00b0] inter\u00adactions and weak aromatic \u03c0\u2013\u03c0 stacking [Cg2\u22efCg2 = 3.9694\u2005(9)\u2005\u00c5], where Cg1 and Cg2 are the centroids of the C3\u2013C8 and C9\u2013C14 rings, respectively = 2.9828\u2005(13)\u2005\u00c5; van der Waals contact distance = 3.37\u2005\u00c5] are observed.In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds into chains propagating parallel to the c axis Table\u00a01. The cryly Fig.\u00a02. In addiCrystalExplorer17 and those delineated into H\u22efH, O\u22efH/H\u22efO, Br\u22efH/H\u22efBr, Br\u22efC/C\u22efBr and F\u22efH/H\u22efF contacts . In the Br\u22efH/H\u22efBr fingerprint plot, there are two symmetrical wings with de + di \u2243 2.85\u2005\u00c5 and they contribute 15.5% to the Hirshfeld surface . The pair of characteristic wings in the fingerprint plot delin\u00adeated into Br\u22efC/C\u22efBr contacts , have the tips at de + di \u2243 3.80\u2005\u00c5, while for F\u22efH/H\u22efF contacts , they have the tips at de + di \u2243 2.60\u2005\u00c5. The remaining contributions from the other different inter\u00adatomic contacts to the Hirshfeld surfaces are listed in Table\u00a03et al., 2015The overall two-dimensional fingerprint plot Fig.\u00a04a and thce Fig.\u00a04c. In thce Fig.\u00a04d. The pet al., 2016E)-1--2-phenyl\u00addiazene unit gave 26 hits. Seven compounds are closely related to the title compound, viz. CSD refcode GUPHIL (I) , mol\u00adecules are linked into inversion dimers via short halogen\u2013halogen contacts [Cl1\u22efCl1 = 3.3763\u2005(9)\u2005\u00c5 C16\u2014Cl1\u22efCl1 = 141.47\u2005(7)\u00b0] compared to the van der Waals radius sum of 3.50\u2005\u00c5 for two chlorine atoms. No other directional contacts could be identified and the shortest aromatic-ring-centroid separation is greater than 5.25\u2005\u00c5. In the crystals of (II) and (III), the aromatic rings form dihedral angles of 64.1\u2005(2) and 60.9\u2005(2)\u00b0, respectively. Mol\u00adecules are linked through weak X\u22efCl contacts [X = Cl for (II) and Br for (III)], C\u2014H\u22efCl and C\u2014Cl\u22ef\u03c0 inter\u00adactions into sheets lying parallel to the ab plane. In the crystal of (IV), the planes of the benzene rings make a dihedral angle of 56.13\u2005(13)\u00b0. Mol\u00adecules are stacked in columns along the a-axis direction via weak C\u2014H\u22efCl hydrogen bonds and face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions. The crystal packing is further consolidated by short Cl\u22efCl contacts. In (V), the benzene rings form a dihedral angle of 63.29\u2005(8)\u00b0. Mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds into zigzag chains running along the c-axis direction. The crystal packing also features C\u2014Cl\u22ef\u03c0, C\u2014F\u22ef\u03c0 and N\u2014O\u22ef\u03c0 inter\u00adactions. In the crystals of (VI) and (VII), the dihedral angles between the aromatic rings are 60.31\u2005(14) and 56.18\u2005(12) \u00b0, respectively. In (VI) C\u2014H\u22efN and short Cl\u22efCl contacts are observed and in (VII), C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and short Cl\u22efO contacts occur.In the crystal of (E)-1-(4-fluoro\u00adphen\u00adyl)-2-(2-nitro\u00adbenzyl\u00adidene)hydrazine (1\u2005mmol), tetra\u00admethyl\u00adethylenedi\u00adamine (TMEDA) , CuCl and CBr4 (4.5\u2005mmol). After 1\u20133h the reaction mixture was poured into a \u223c0.01 M solution of HCl , and extracted with di\u00adchloro\u00admethane (3 \u00d7 20\u2005ml). The combined organic phase was washed with water (3 \u00d7 50\u2005ml), brine (30\u2005ml), dried over anhydrous Na2SO4 and concentrated in vacuo using a rotary evaporator. The residue was purified by column chromatography on silica gel using appropriate mixtures of hexane and di\u00adchloro\u00admethane (3/1\u20131/1). Crystals suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution. Light-orange solid (52%); m.p. 377\u2005K. Analysis calculated for C14H8Br2FN3O2 (M = 429.04): C 39.19, H 1.88, N 9.79; found: C 39.14, H 1.87, N 9.73%. 1H NMR \u03b4 7.86\u20137.14 . 13C NMR \u03b4 165.02, 163.23, 163.01, 149.72, 133.01, 132.10, 129.70, 124.98, 124.87, 124.80, 124.29, 116.07, 115.91, 86.88. ESI\u2013MS: m/z: 430.02 [M + H]+.A 20\u2005ml screw-neck vial was charged with DMSO (10\u2005ml), (Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S205698902200278X/hb8012sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902200278X/hb8012Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902200278X/hb8012Isup3.cmlSupporting information file. DOI: 2158375CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Five new bis\u00ad(aryl\u00adamide)\u00addichlorido\u00adzinc(II) complexes have been prepared and characterized. All of the complexes contain hydrogen bonds between the amide N\u2014H group and the amide carbonyl oxygen atoms or the chlorine atoms, forming extended networks. O)di\u00adchlorido\u00adzinc(II), [ZnCl2(C7H7NO)2], di\u00adchlor\u00adido\u00adbis\u00ad(2-methyl\u00adbenzamide-\u03baO)zinc(II), [ZnCl2(C8H9NO)2], di\u00adchlorido\u00adbis\u00ad(3-methyl\u00adbenzamide-\u03baO)zinc(II), [ZnCl2(C8H9NO)2], di\u00adchlorido\u00adbis\u00ad(4-methyl\u00adbenzamide-\u03baO)zinc(II), [ZnCl2(C8H9NO)2], and di\u00adchlorido\u00adbis\u00ad(4-hy\u00addroxy\u00adbenzamide-\u03baO)zinc(II), [ZnCl2(C7H7NO2)2]. All of the complexes contain hydrogen bonds between the amide N\u2014H group and the amide carbonyl oxygen atoms or the chlorine atoms, forming extended networks.Ionic co-crystals are co-crystals between organic mol\u00adecules and inorganic salt coformers. Co-crystals of pharmaceuticals are of inter\u00adest to help control polymorph formation and potentially improve stability and other physical properties. We describe the preparation, crystal structures, and hydrogen bonding of five different 2:1 benzamide or tolu\u00adamide/zinc(II) chloride co-crystal salts, namely, bis\u00ad(benzamide-\u03ba All five complexes are 2:1 O-bonded aryl amide:ZnCl2 complexes with approximately tetra\u00adhedral zinc(II) centers. The complexes crystallize in five different space groups and form hydrogen-bonding inter\u00adactions between the amide N\u2014H groups and either an amide oxygen or a zinc-bound chlorido ligand.Five new zinc complexes, (1), bis\u00ad(benzamide-\u03baO)di\u00adchlorido\u00adzinc(II), [ZnCl2(C7H7NO)2], crystallizes in the P21/n space group with two independent mol\u00adecules in the asymmetric unit and displays one N\u2014H\u22efO and one N\u2014H\u22efCl intra\u00admolecular hydrogen bond in each mol\u00adecule , di\u00adchlorido\u00adbis\u00ad(2-methyl\u00adbenzamide-\u03baO)zinc(II), [ZnCl2(C8H9NO)2], displays two intra\u00admolecular N\u2014H\u22efCl hydrogen bonds to one chlorine atom (see Table\u00a02P21 space group. Compound (3), di\u00adchlorido\u00adbis\u00ad(3-methyl\u00adbenzamide-\u03baO)zinc(II), [ZnCl2(C8H9NO)2], crystallizes in the C2/c space group with the zinc atom lying on the twofold axis does not form any intra\u00admolecular hydrogen bonds. Compound (4), di\u00adchlorido\u00adbis\u00ad(4-methyl\u00adbenzamide-\u03baO)zinc(II), [ZnCl2(C8H9NO)2], crystallizes in the P21/c space group and compound (5), di\u00adchlorido\u00adbis\u00ad(4-hy\u00addroxy\u00adbenzamide-\u03baO)zinc(II), [ZnCl2(C7H7NO2)2], crystallizes in the Cc space group and both compounds form two intra\u00admolecular hydrogen bonds, one N\u2014H\u22efO and one N\u2014H\u22efCl, similar to the inter\u00adactions found in compound (1) found in a search of the CSD forms four N\u2014H\u22efCl inter\u00admolecular hydrogen bonds (two from each independent mol\u00adecule), forming an extended network as shown in Fig.\u00a062) also utilizes N\u2014H bonds in hydrogen-bonding inter\u00adactions, two intra\u00admolecular and two inter\u00admolecular, to form layers within the structure forms two N\u2014H\u22efCl inter\u00admolecular contacts in addition to the two intra\u00admolecular hydrogen bonds, resulting in a complex set of layers that run perpendicular to the b axis results in the greatest number of hydrogen bonds among this set of complexes, as shown in Fig.\u00a010Each compound displays a unique hydrogen-bonding network, consisting primarily of N\u2014H\u22efO and N\u2014H\u22efCl inter\u00adactions, summarized in Table\u00a011), (3), and (5) form \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of the benzamide or tolu\u00adamide groups as summarized in Table\u00a072) or (4).Compounds . The similar di\u00adchlorido\u00adbis\u00ad(dmf)zinc(II) has also been reported. Edwards et al. , the latter of which undergoes a reversible phase transition at 228\u2005K 2 where X = Cl, Br, I . As part of a larger study, Smirnov et al. , thio\u00adacetamide (VUCTOD), and benzamide (VUCVAR).A number of zinc(II) iodide complexes, ZnI al. 2009 have alset al. , one water molecule, and two chlorido ligands. In the second form (CCDC 2010264), the zinc bonds to one nefiracetam molecule through the \u03b3-lactam and to a second via the amide carbonyl, forming a cyclic zinc dimer.Three structural studies have prepared zinc(II)chloride complexes with pharmaceutically relevant mol\u00adecules. Sultana al. 2016 prepared al. 2020 describe1) through (5) were prepared by dissolution of the respective components in various solvents followed by slow evaporation. In a typical preparation, a 1:1 stoichiometric ratio of benzamide (0.1352\u2005g) and zinc(II) chloride (0.1336\u2005g) was dissolved in approximately 5\u2005mL of a 50:50 v:v ratio of water and ethanol. Slow evaporation of the resulting solution produced single crystals of compound (1).All reagents were used as received from the manufacturer. Compounds = 1.2Ueq(C) (aromatic) or Uiso(H) = 1.5Ueq(C) (meth\u00adyl). All amide H-atom positions were refined with N\u2014H distances restrained to 0.88\u2005(2)\u2005\u00c5 and Uiso(H) = 1.5Ueq(N). The hydroxyl H-atom positions in compound (5) were refined with O\u2014H distances restrained to 0.84\u2005(2)\u2005\u00c5 and Uiso(H) = 1.5Ueq(N).All carbon-bonded H atoms were placed in idealized positions using a riding model with aromatic C\u2014H = 0.95\u2005\u00c5, methyl C\u2014H = 0.98\u2005\u00c5 and 1) was refined as a pseudo-merohedral twin with a twin law of (0 0 \u22121 0 \u22121 0 \u22121 0 0) , corresponding to a twofold rotation about the [10Compound (3) was modeled as a disordered methyl group with each set of hydrogen atoms rotated by 60\u00b0 (AFIX 127). The disorder was identified from multiple peaks near C8 in the difference map. The refined occupancies of the two hydrogen atom sets were 0.54\u2005(2):0.46\u2005(2).The methyl group in compound global, 1, 2, 3, 4, 5. DOI: 10.1107/S2056989021008264/zl50231sup2.hklStructure factors: contains datablock(s) 1. DOI: Click here for additional data file.10.1107/S2056989021008264/zl50231sup7.molSupporting information file. DOI: 10.1107/S2056989021008264/zl50232sup3.hklStructure factors: contains datablock(s) 2. DOI: Click here for additional data file.10.1107/S2056989021008264/zl50232sup8.molSupporting information file. DOI: 10.1107/S2056989021008264/zl50233sup4.hklStructure factors: contains datablock(s) 3. DOI: Click here for additional data file.10.1107/S2056989021008264/zl50233sup9.molSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021008264/zl50234sup10.molSupporting information file. DOI: 10.1107/S2056989021008264/zl50234sup5.hklStructure factors: contains datablock(s) 4. DOI: Click here for additional data file.10.1107/S2056989021008264/zl50235sup11.molSupporting information file. DOI: 10.1107/S2056989021008264/zl50235sup6.hklStructure factors: contains datablock(s) 5. DOI: 2102513, 2102512, 2102511, 2102510, 2102509CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II featuring piano-stool geometries and supported by tri\u00adaryl\u00adphosphine ligands, showing the effects of para substitution on supra\u00admolecular structure and allowing comparison to the large class of previously reported acetyl complexes.Solid-state structures are presented for three propionyl complexes of Mo para substituents, namely, dicarbon\u00adyl(\u03b75-cyclo\u00adpenta\u00addien\u00adyl)propion\u00adyl(tri\u00adphenyl\u00adphosphane-\u03baP)molybdenum(II), [Mo(C5H5)(C3H5O)(C18H15P)(CO)2], (1), dicarbon\u00adyl(\u03b75-cyclo\u00adpenta\u00addien\u00adyl)propion\u00adyl[tris\u00ad(4-fluoro\u00adphen\u00adyl)phosphane-\u03baP]molybdenum(II), [Mo(C5H5)(C3H5O)(C18H12F3P)(CO)2], (2), and dicarbon\u00adyl(\u03b75-cyclo\u00adpenta\u00addien\u00adyl)propion\u00adyl[tris\u00ad(4-meth\u00adoxy\u00adphen\u00adyl)phosphane-\u03baP]molybdenum(II) dichloromethane solvate, [Mo(C5H5)(C3H5O)(C21H21O3P)(CO)2]\u00b7CH2Cl2, (3), have been prepared from the corresponding ethyl complexes via phosphine-induced migratory insertion. These complexes exhibit four-legged piano-stool geom\u00adetries with mol\u00adecular structures quite similar to each other and to related acetyl complexes. The extended structures of the three complexes differ somewhat, with the para substituent of the tri\u00adaryl\u00adphosphine of (2) (fluoro) or (3) (meth\u00adoxy) engaging in non-classical C\u2014H\u22efF or C\u2014H\u22efO hydrogen-bonding inter\u00adactions. The structure of (3) exhibits modest disorder in the position of one Cl atom of the di\u00adchloro\u00admethane solvent, which was modeled with two sites showing approximately equivalent occupancies [0.532\u2005(15) and 0.478\u2005(15)].Three cyclo\u00adpenta\u00addienylmolybdenum(II) propionyl complexes featuring tri\u00adaryl\u00adphosphine ligands with different Although substitution of phosphine aryl groups with electron-withdrawing or -donating groups minimally affects local structure, the supra\u00admolecular organization is substanti\u00adally affected by non-classical hydrogen-bonding to the fluoro and meth\u00adoxy groups in (2) and (3), respectively.We were inter\u00adested in extending earlier studies to higher-order alkyl groups at molybdenum, and in this report we describe the synthesis and solid-state structures of related Mo1), (2), and (3) are illustrated in Figs. 1trans-disposed carbonyl ligands. As previously observed for most related acetyl complexes, the acyl C=O points up toward the Cp ring. In the case of (1), this phenomenon could be rationalized by presence of short C4\u2014H4A\u22efO1 (2.672\u2005\u00c5) and C4\u2014H4B\u22efO2 (2.639\u2005\u00c5) contacts involving the carbonyl ligands that are enabled when the acyl points up. However, the variation of the Mo1\u2014C3\u2014C4\u2014C5 torsion angle across the series argues against the general importance of such an inter\u00adaction.The mol\u00adecular structures of (1), (2), and (3) are presented in Tables 12) crystallized with two nearly equivalent mol\u00adecules in the asymmetric unit, so geometric parameters are presented for both. In general, the three complexes are nearly identical, as might be expected based on the dominant role of sterics in determining structure and the fact that the steric profiles of the three tri\u00adaryl\u00adphosphine ligands are identical. The Mo\u2014P bond length in (2) [2.4692\u2005(4)\u2005\u00c5 (avg)] is slightly shorter than in (1) or (3) , which may be related to stronger \u03c0-backbonding to the tris\u00ad(4-fluoro\u00adphen\u00adyl)phosphine ligand. Stronger backbonding is supported by the observation by infrared spectroscopy of slightly higher-energy carbonyl stretching vibrations for (2) [\u03bd(CO)avg = 1897\u2005cm\u22121] compared with (1) and (3) . Geometric parameters for all complexes are quite similar to those for the related tri\u00adphenyl\u00adphosphine-supported CpMo acetyl complex , (2), and (3) in their mol\u00adecular structures, the para substituent of the tri\u00adaryl\u00adphosphine ligand plays an important role in determining the extended structure. The extended structure of (1) is dominated by non-classical C\u2014H\u22efO inter\u00adactions involving its carbonyl ligands. A short C\u2014H\u22efO inter\u00adaction between O2 and H12 of a phenyl ring (2.36\u2005\u00c5) joins mol\u00adecules of (1) into centrosymmetrical dimers that are organized into chains along [010] by inter\u00admolecular C15\u2014H15\u22efCg4 and intra\u00admolecular O2\u22efCg4 (3.295\u2005\u00c5) inter\u00adactions features two nearly equivalent mol\u00adecules in the asymmetric unit exhibiting a non-classical C\u2014H\u22efO inter\u00adaction between O6 of a propionyl ligand and H15 from a phenyl ring (2.59\u2005\u00c5) and a C\u2014H\u22efF close contact between F3 and H53 (2.60\u2005\u00c5). These pairs of mol\u00adecules are joined into chains along [001] by C34\u2014H34\u22efO1 hydrogen bonding .The tris\u00ad(4-fluoro\u00adphen\u00adyl)phosphine-supported derivative (1), complex (3) is joined into centrosymmetrical dimers by a C\u2014H\u22efO inter\u00adaction involving a carbonyl ligand . This further set of inter\u00adactions involving meth\u00adoxy groups, as well as important close contacts involving the di\u00adchloro\u00admethane solvent, are depicted in Fig.\u00a09Like complex (CO)2(PR3)(COR). The trans-dicarbonyl structure, as observed for (1)\u2013(3), is preferred except in cases where the phosphine and acyl ligands are covalently linked, forcing them to be cis CpMo(CO). This compound was prepared by modification of the method used of Gladysz et al. was dissolved in THF (10\u2005ml). Sodium tri\u00adethyl\u00adborohydride was added dropwise by syringe with vigorous stirring, leading to an immediate color change from purple to green\u2013yellow with evolution of H2 gas. The reaction was allowed to proceed with stirring for 20\u2005min, and an excess of iodo\u00adethane was added dropwise with stirring and the reaction was allowed to proceed for 6\u2005h. Volatiles were removed in vacuo to afford a yellow\u2013brown film that was stored at 238\u2005K for 1 week. The solid was extracted with pentane (4 \u00d7 10\u2005ml) and filtered through a 1\u2005cm pad of activated alumina to afford a yellow solution, and removal of solvent in vacuo afforded 3(CH2CH3)CpMo(CO) as a pure yellow powder . 1H NMR : \u03b4 5.28 , 1.72 , 1.45 . 13C{1H} NMR : \u03b4 239.9 (Mo\u2014CO), 227.8 (Mo\u2014CO), 93.0 (Cp ring), 20.4 (Mo\u2014CH2CH3) \u22123.7 (Mo\u2014CH2CH3). IR \u03bd(CO): 2015, 1921 (split). al. 1979, as prev al. 1979 and Anst al. 2020. In a 202(PPh3)(COCH2CH3) (1)CpMo(CO). In an inert-atmos\u00adphere glove box, CpMo(CO)3(CH2CH3) and tri\u00adphenyl\u00adphosphine were dissolved in aceto\u00adnitrile (5\u2005ml) in a 20\u2005ml scintillation vial equipped with a flea-sized stir bar. The mixture was stirred for 1 week, during which time a bright-yellow precip\u00aditate formed. The yellow solid was isolated by filtration and washed with pentane (2 \u00d7 5\u2005ml), then dried in vacuo to afford pure 1. Yellow crystals of 1 suitable for X-ray diffraction were obtained from a concentrated di\u00adchloro\u00admethane solution by vapor cross diffusion with pentane at 238\u2005K. 1H NMR : \u03b4 7.50\u20137.30 , 5.00 , 3.03 CH2CH3), 0.90 CH2CH3). 13C{1H} NMR : \u03b4 267.7 , 238.8 , 135.7 , 133.2 , 130.5 , 128.6 , 96.7 (Cp ring), 58.1 (Mo\u2014COCH2CH3), 10.1 (Mo\u2014COCH2CH3). 31P{1H} NMR : \u03b4 68.4 (s). IR \u03bd(CO): 1935, 1851, 1614 (acet\u00adyl).2(P(4-FPh)3)(COCH2CH3) (2)CpMo(CO). In an inert-atmosphere glove box, CpMo(CO)3(CH2CH3) and tris\u00ad(4-fluoro\u00adphen\u00adyl)phosphine were dissolved in aceto\u00adnitrile (5\u2005ml) in a 20\u2005ml scintillation vial equipped with a flea-sized stir bar. The mixture was stirred for 1 week, causing a color change to orange, but without formation of any precipitate. Solvent was removed in vacuo, causing precipitation of a yellow solid that was isolated by filtration and washed with pentane (2 \u00d7 3\u2005ml) to afford the desired product 2 . Yellow crystals of 2 suitable for X-ray diffraction were obtained from a concentrated di\u00adchloro\u00admethane solution by vapor cross diffusion with pentane at 238\u2005K. 1H NMR : \u03b4 7.41\u20137.30 , 7.14 , 4.90 , 2.99 CH2CH3), 0.90 CH2CH3). 13C{1H} NMR : \u03b4 265.4 , 238.2 , 164.0 , 135.0 , 131.3 , 116.0 , 96.5 (Cp ring), 58.2 (Mo\u2014COCH2CH3), 10.9 (Mo\u2014COCH2CH3). 31P{1H} NMR : \u03b4 68.5 (s). IR \u03bd(CO): 1938, 1856, 1620 (acet\u00adyl).2(P(4-MeOPh)3)(COCH2CH3) (3)CpMo(CO). In an inert-atmosphere glove box, CpMo(CO)3(CH2CH3) and tris\u00ad(4-meth\u00adoxy\u00adphen\u00adyl)phosphine were dissolved in aceto\u00adnitrile (5\u2005ml) in a 20 nml scintillation vial equipped with a flea-sized stir bar. The mixture was stirred for 1 week, causing precipitation of 3 as a pure yellow powder that was isolated by filtration. Crystals of 3 suitable for X-ray diffraction were obtained from a concentrated di\u00adchloro\u00admethane solution by vapor cross diffusion with pentane at 238\u2005K. 1H NMR : \u03b4 7.37\u20137.23 , 7.14 , 4.99 , 3.03 CH2CH3), 0.89 CH2CH3). 13C{1H} NMR : \u03b4 268.6 , 239.2 , 161.1 (C\u2014OCH3 of phosphine), 134.6 , 127.4 , 114.0 , 96.6 (Cp ring), 58.0 (Mo\u2014COCH2CH3), 10.1 (Mo\u2014COCH2CH3). 31P{1H} NMR : \u03b4 62.3 (s). IR \u03bd(CO): 1933, 1847, 1605 (acet\u00adyl).Uiso(H) = k\u00d7Ueq(C), k = 1.2 for cyclo\u00adpenta\u00addienyl, phenyl, and methyl\u00adene groups and 1.5 for methyl groups. Methyl group H atoms were allowed to rotate in order to find the best rotameric conformation.Crystal data, data collection and structure refinement details are summarized in Table\u00a071); seven for (2); five for (3)] are missing from these high-quality data sets due to the arrangement of the instrument with a conservatively sized beam stop. The large number of reflections in the data sets (and the Fourier-transform relationship of intensities to atoms) ensures that no particular bias has been introduced.A small number of intense low-angle reflections .The structure of global, 1, 2, 3. DOI: 10.1107/S2056989021008008/jq20081sup2.hklStructure factors: contains datablock(s) 1. DOI: Click here for additional data file.10.1107/S2056989021008008/jq20081sup5.cdxSupporting information file. DOI: 10.1107/S2056989021008008/jq20082sup3.hklStructure factors: contains datablock(s) 2. DOI: Click here for additional data file.10.1107/S2056989021008008/jq20082sup6.cdxSupporting information file. DOI: 10.1107/S2056989021008008/jq20083sup4.hklStructure factors: contains datablock(s) 3. DOI: Click here for additional data file.10.1107/S2056989021008008/jq20083sup7.cdxSupporting information file. DOI: 2101246, 2101245, 2101244CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The \u03b2-tubulin blot in Figure\u00a05B (middle panel) was inadvertently taken from Figure\u00a05F (MEK1 blot in the third panel) while preparing the figures. The amended Figure\u00a05B (middle panel) here shows the correct \u03b2-tubulin blot."} +{"text": "In the title compound,the two pyridine side arms are not coplanar, with the terminal pyridine rings subtending a dihedral angle of 26.45\u2005(6)\u00b0. In the crystal, hydrogen bonds, inter\u00admolecular C\u2014H\u22efCl contacts and a weak C\u2014H\u22efO inter\u00adaction connect the mol\u00adecule with neighbouring chloride counter-anions and lattice water mol\u00adecules. The crystal packing also features by \u03c0\u2013\u03c0 inter\u00adactions. 19H23N52+\u00b72Cl\u2212\u00b72H2O, the two pyridine side arms are not coplanar, with the terminal pyridine rings subtending a dihedral angle of 26.45\u2005(6)\u00b0. In the crystal, hydrogen bonds, inter\u00admolecular C\u2014H\u22efCl contacts and a weak C\u2014H\u22efO inter\u00adaction connect the mol\u00adecule with neighbouring chloride counter-anions and lattice water mol\u00adecules. The crystal packing also features by \u03c0\u2013\u03c0 inter\u00adactions with centroid-centroid distances of 3.4864\u2005(12) and 3.5129\u2005(13)\u2005\u00c5.In the title compound, C Bond leIn the crystal, there are inter\u00admolecular hydrogen bonds Table\u00a01 and C\u2014H\u22efa-axis direction with a Cg1\u22efCg2 centroid\u2013centroid distance of 3.4864\u2005(12)\u2005\u00c5, a perpendicular distance from the centroid Cg1 to the plane of the other ring of 3.2472\u2005(8)\u2005\u00c5 and a slippage between the centroids of 1.100\u2005\u00c5. Similarly, the second \u03c0\u2013\u03c0 stacking inter\u00adaction has a Cg3\u22efCg3 centroid-centroid distance of 3.5129\u2005(13)\u2005\u00c5, a perpendicular distance from the centroid Cg3 to the plane of the other ring of 3.2177\u2005(8)\u2005\u00c5 and a slippage between the centroids of 1.410\u2005\u00c5. Cg1, Cg2 and Cg3 are the centroids of N1/C8\u2013C12, N3/C1\u2013C5 and N5/C15\u2013C19 pyridine rings, respectively pyridine-2,6-dicarboxamide meth\u00adyl]pyridine-2,6-dicarboxamide hemihydrate methan\u00adam\u00adinium] dichloride dihydrate compound was obtained following the procedure previously reported in the literature (tos\u00adyl)amino]\u00admeth\u00adyl}pyridine, which could be isolated after chromatography bis\u00ad[Slow diffusion between toluene and a wet di\u00adchloro\u00admethane solution of the brown oil set aside at room temperature gave colourless needles of the title compound suitable for X-ray diffraction within five days.Uiso(H) = 1.2Ueq(C). For two similar N\u2014H distances, a restraint was applied to make them approximately equal with an effective standard deviation of 0.02\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698902101183X/tx2045sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902101183X/tx2045Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902101183X/tx2045Isup3.cmlSupporting information file. DOI: 2120892CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Z configuration. In the crystal, the mol\u00adecules are linked by weak C\u2014H\u22ef\u03c0 hydrogen bonds and very weak \u03c0\u2013\u03c0 stacking inter\u00adactions.The title Schiff base exists in the enol\u2013imine tautomeric form and adopts a 16H17NO, is a Schiff base that exists in the enol\u2013imine tautomeric form and adopts a Z configuration. The mol\u00adecule is non-planar, with the twisted rings making a dihedral angle of 39.92\u2005(4)\u00b0. The intra\u00admolecular O\u2014H\u22efN hydrogen bond forms an S(6) ring motif. In the crystal, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions and very weak \u03c0-\u03c0 stacking inter\u00adactions also help to consolidate the crystal packing. A Hirshfeld surface analysis was performed to investigate the contributions of different inter\u00admolecular contacts within the supra\u00admolecular structure. The major contributions are from H\u22efH (65%), C\u22efH (19.2%) and O\u22efH (6.6%) inter\u00adactions.The title compound, C These compounds can be easily synthesized by condensation of a primary aliphatic or aromatic amine with an aldehyde or ketone in different solvent media and they can easily be purified, since the amount of by-products is negligible ring motif. The C1\u2014O1 [1.353\u2005(2)\u2005\u00c5] and C7\u2014N1 [1.282\u2005(2)\u2005\u00c5] bond distances indicate their single- and double-bond characters, respectively, being consistent with the phenol\u2013imine tautomeric form.The title compound crystallizes in the phenol\u2013imine tautomeric form with an le Fig.\u00a01, which iA\u22ef\u03c0 (C9\u2013C14) inter\u00adactions = 4.0220\u2005(9)\u2005\u00c5] lead to additional stabilization of the crystal packing. A view of the crystal packing parallel to the bc plane is shown in Fig.\u00a02In the crystal, mol\u00adecules are linked by C16\u2014H16s Table\u00a01, and veret al., 2016Z)-2-{[imino]\u00admeth\u00adyl}-4-methyl\u00adphenol unit, revealed ten hits where this fragment adopts the enol\u2013imine tautomeric form. The imine bond length (N1\u2014C7) in the title compound is the same within standard uncertainties as the corresponding bond lengths in the structures of 2-(di\u00adphenyl\u00admeth\u00adyl)-6-[(mesityl\u00adimino)\u00admeth\u00adyl]-4-methyl\u00adphenol -N,N\u2032-bis\u00ad-5,5\u2032,6,6\u2032,7,7\u2032,8,8\u2032-octa\u00adhydro-1,1\u2032-binaphthyl-2,2\u2032-di\u00adamine \u00admeth\u00adyl]phen\u00ado\u00adlato}tetra\u00adhydro\u00adfuran\u00admagnesium imino]\u00admeth\u00adyl}phenol to 1.1632\u2005\u00c5 (blue) . In Figs. 2normd and ed surfaces represent the C\u2014H\u22efCg inter\u00adactions. The most important inter\u00adaction is H\u22efH, contributing 65% to the overall crystal packing, which is illustrated in the 2D fingerprint -2-{[imino]\u00admeth\u00adyl}-4-methyl\u00adphenol was synthesized by condensation of 2-hy\u00addroxy-5-methyl\u00adbenzaldehyde and 2,4-di\u00admethyl\u00adaniline = 1.5Ueq(O). The C-bound H atoms were positioned geometrically and refined using a riding model with C\u2014H = 0.93 and Uiso(H) = 1.2Ueq(C) for sp2-hybridized C atoms and with C\u2014H = 0.96\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021010215/yk2157sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021010215/yk2157Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021010215/yk2157Isup3.cmlSupporting information file. DOI: 2113562CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Their structures with absolute configurations were elucidated by extensive spectroscopic methods. Compounds 1\u20133 showed antibacterial activity against four strains with MIC values in a range of 16\u2013128\u00a0\u03bcg/mL.Five previously undescribed guanacastane diterpenoids, namely psathyrellins A\u2013E (The online version contains supplementary material available at 10.1007/s13659-021-00316-x. Psathyrella candolleana (Psathyrellaceae) is a small agaric usually found in the vicinity of recently dead hardwood trees. It has a wide distribution on lawns or pastures in Europe and North America. Previous pharmacological studies indicated that the extracts of P. candolleana showed a marked anticlastogenic effect against DNA damage + . The UV data at 282\u00a0nm suggested a conjugated system in 2. The 1D and 2D NMR spectra revealed similar patterns to those of 1 except that one more double bond in 2. In the HMBC spectrum, correlations from \u03b4H 1.11 to \u03b4C 142.8 and 158.6 , from \u03b4H 6.99 to C-1 and \u03b4C 158.6 , and from \u03b4H 1.74 to C-3 and \u03b4C 131.5 suggested that three double bonds were distributed at C-12/C-13, C-1/C-2, and C-3/C-4, respectively. Detailed analysis of 2D NMR data suggested that the other parts of 2 were the same to those of 1. A single crystal X-ray diffraction not only proved the planar structure, but also determined the absolute configuration of 1 as shown in Fig.\u00a0Psathyrellin B (3) was isolated as a yellow oil. The molecular formula was determined as C20H28O5 on the basis of HRESIMS data at m/z 349.20093 [M\u2009+\u2009H]+ . All NMR data suggested that 3 was structurally similar to that of 2 was isolated as a yellow oil. The molecular formula was determined as C22H28O6 on the basis of HRESIMS data at m/z 411.17749 [M\u2009+\u2009Na]+ . All spectroscopic data of 4 were similar to those of 2 excepted two additional carbons at \u03b4C 20.8 (q) and 172.8 (s) in 4 that were easily assigned as an O-acetyl moiety. The HMBC correlations from \u03b4H 4.40 and 4.30 to \u03b4C 172.8 (s) and from \u03b4H 1.27 to \u03b4C 32.3 , 67.1 suggested that the O-acetyl moiety should be placed at C-19. Detailed analysis of 2D NMR data suggested that the other parts of 4 were the same to those of 2. The CD spectrum almost showed the same curve to that of 2, indicating the absolute configuration of the main backbone in 4 to be the same to that of 2 was isolated as a yellow oil. The molecular formula was determined as C22H28O5 on the basis of HRESIMS data at m/z 373.20084 [M\u2009+\u2009H]+ . According to analysis of 1D and 2D NMR data, compound 5 was easily identified as a 19-O-acetyl derivative of 2, which was also similar to 4. The only difference was that the hydroxymethine of C-6 in 4 was reduced into a methylene at \u03b4C 34.5 in 5, as supported by the HMBC correlation from H-6 to C-5 and the 1H\u20131H COSY data between H-6 and H-7. The CD spectrum almost indicated the same curve to that of 2, indicating the absolute configuration of the backbone in 5 to be the same to that of 2 and a Dionex UltiMate 3000 RSLC UPLC system. Silica gel (200\u2013300 mesh and 500\u2013800 mesh), RP-18 gel (40\u201375\u00a0\u00b5m) and Sephadex LH-20 were used for column chromatography (CC). Preparative HPLC was performed on an Agilent 1260 liquid chromatography system with a Zorbax SB-C18 column, a Daicel chiral column and a DAD detector.Optical rotations were measured on a Rudolph Autopol IV polarimeter. UV spectra were obtained on a UH5300 UV\u2013VIS Double Beam Spectrophotometer. IR spectra were obtained by using a Shimadzu Fourier Transform Infrared spectrometer with KBr pellets. NMR spectra were acquired with a Bruker Avance III 600 instrument. CD spectra were recorded with an Applied Photophysics Chirascan-Plus spectrometer. High resolution electrospray ionization mass spectra (HRESIMS) were recorded on a LC\u2013MS system consisting of a Q Exactive\u2122 Orbitrap mass spectrometer with an ESI ion source used in ultra-high resolution mode was deposited at School of Pharmaceutical Sciences, South-Central University for Nationalities. The strains were cultured in PDA and stored at \u2212\u00a04\u00a0\u00b0C. Culture medium was composed of glucose (5%), pork pepton (0.15%), yeast (0.5%), KH2PO4 (0.05%) and MgSO4 (0.05%). Initial pH was adjusted to 6.0, the fermentation was first carried out on an erlenmeyer flask for 6\u00a0days till the mycelium biomass reached to the maximum. Then it was transferred to rice medium at 24\u00a0\u00b0C in dark culture for 40\u00a0days. The rice medium in each 250\u00a0mL-Erlenmeyer flask was composed of rice (50\u00a0g) and water (50\u00a0mL). A total of 180 bottles were used in this study.Fruiting bodies of 2CO (from 20:1 to 1:1) to afford eight fractions A\u2013H. Fraction C (3.8\u00a0g) was isolated by CC over silica gel using PE/Me2CO (6/1) to give subfractions C1\u2013C6. Compound 1 was deposited from fraction C4 as colorless crystals . Fraction E (4\u00a0g) was first isolated by silica gel CC (200\u2013300 mesh) eluted with PE/Me2CO (5/1) to give five subfractions E1-E5. Fraction E2 (800\u00a0mg) was further isolated by CC using RP-C18 silica gel (MeOH/H2O from 6/4 to 9/1) to give subfractions E2a-E2e. HPLC preparation (MeCN/H2O from 7/3 to 8/2 in 20\u00a0min) on fraction E2d (82\u00a0mg) afforded compounds 3 \u2009=\u200912.1\u00a0min; purity 90%), 4 , and 2 . Fraction E2e (70\u00a0mg) was separated by CC over Sephadex LH-20 (MeOH) to give a mixture. The mixture was subjected to HPLC (MeCN/H2O from 7/3 to 8/2 in 20\u00a0min) to give compound 5 .The rice fermentation (9\u00a0kg) was extracted four times with EtOAc. The organic layer was evaporated to give a crude extract (90\u00a0g). The extract was subjected to silica gel CC (200\u2013300 mesh) eluted with a gradient solvent system of petroleum ether (PE)/Me\u03b1]D15 \u2013 208.1 ; UV (MeOH) \u03bbmax (log \u03b5) 192 (3.36), 244 (3.18) nm; IR (KBr) \u03bdmax 3421, 3349, 2829, 1710, 1702, 1640, 1462, 1038\u00a0cm\u22121; 1H (600\u00a0MHz) and 13C NMR (150\u00a0MHz) data (methanol-d4), see Table m/z 333.20584 [M\u2009+\u2009H]+ .Colorless crystals (MeOH); [\u03b1]D15 \u2013 239.2 ; UV (H2O) \u03bbmax (log \u03b5) 190 (3.34), 252 (3.20), 282 (2.86) nm; ECD (MeOH) \u03bbmax\u00a0(\u0394\u03b5) 223 (+49), 251 (+45), 287 (\u201339), 339 (\u201327) nm;\u00a0IR (KBr) \u03bdmax 3432, 3346, 2910, 1711, 1706, 1642, 1446, 1036\u00a0cm\u22121; 1H (600\u00a0MHz) and 13C NMR (150\u00a0MHz) data (methanol-d4), see Table m/z 331.19031 [M\u2009+\u2009H]+ .Colorless crystals (MeOH); [\u03b1]D15 \u2013 182.9 ;\u00a0ECD (MeOH) \u03bbmax\u00a0(\u0394\u03b5) 209\u00a0(\u201333), 224\u00a0(+26), 287 (\u201329), 341\u00a0(\u20138) nm; 1H (600\u00a0MHz) and 13C NMR (150\u00a0MHz) data (methanol-d4), see Table m/z 349.20093 [M\u2009+\u2009H]+ .Yellow oil; [\u03b1]D15 \u2013 221.9 ; ECD (MeOH) \u03bbmax\u00a0(\u0394\u03b5) 222\u00a0(+20), 243 (+22), 287 (\u201315), 338\u00a0(\u201312) nm;\u00a01H (600\u00a0MHz) and 13C NMR (150\u00a0MHz) data (methanol-d4), see Table m/z 411.17749 [M\u2009+\u2009Na]+ .Yellow oil; [\u03b1]D15 \u2013 292.4 ;\u00a0ECD (MeOH) \u03bbmax\u00a0(\u0394\u03b5) 221\u00a0(+27), 242\u00a0(+31), 290\u00a0(\u201324), 338\u00a0(\u201318) nm; 1H (600\u00a0MHz) and 13C NMR (150\u00a0MHz) data (methanol-d4), see Table m/z 373.20084 [M\u2009+\u2009H]+ .Yellow oil; . The final wR(F2) values were 0.0799 [I\u2009>\u20092\u03c3(I)]. The final R1 values were 0.0307 . The final wR(F2) values were 0.0800 . The goodness of fit on F2 was 1.056. Flack parameter\u2009=\u20090.05(3). CCDC: 2068966 (www.ccdc.cam.ac.uk).C20H26O4, M\u2009=\u2009330.41, a\u2009=\u20098.0026(8) \u00c5, b\u2009=\u20098.3729(9) \u00c5, c\u2009=\u200914.4078(15) \u00c5, \u03b1\u2009=\u200990.00\u00b0, \u03b2\u2009=\u2009105.783(3)\u00b0, \u03b3\u2009=\u200990.00\u00b0, V\u2009=\u2009929.00(17) \u00c53, T\u2009=\u2009150.(2) K, space group P1211, Z\u2009=\u20092, \u03bc(CuK\u03b1)\u2009=\u20091.54178\u00a0mm\u22121, 12059 reflections measured, 3552 independent reflections (Rint\u2009=\u20090.0456). The final R1 values were 0.0559 [I\u2009>\u20092\u03c3(I)]. The final R1 values were 0.0415 . The final wR(F2) values were 0.1156 . The goodness of fit on F2 was 1.092. Flack parameter\u2009=\u2009\u2212\u00a00.12(14). CCDC: 2068967 (www.ccdc.cam.ac.uk).CEscherichia coli ATCC25922, Staphylococcus aureus subsp. aureus ATCC29213, Salmonella enterica subsp. enterica ATCC14028, Pseudomonas aeruginosa ATCC27853 were purchased from China General Microbiological Culture Collection Center, (CGMCC). All these strains were cultured in Mueller Hinton broth (MHB) at 37\u00a0\u00b0C overnight with shaking (200\u00a0rpm). A sample of each culture was then diluted 40-fold in fresh MHB broth and incubated with shaking (200\u00a0rpm) at 37\u00a0\u00b0C for 2.5\u00a0h. The resultant mid-log phase cultures were diluted to a concentration of 5\u2009\u00d7\u2009105\u00a0CFU/mL, then 50\u00a0mL was added to each well of the compound-containing plates. The minimum inhibition concentration (MIC) was determined by measuring bacterial growth after 24\u00a0h on performing 1:2 serial dilutions of each compound ranging from 1 to 128\u00a0\u03bcg/mL. Chloramphenicol was used as a positive control.The tested bacteria strains Supplementary file1 (PDF 2820 KB)Supplementary file2 (CIF 1031 KB)Supplementary file3 (CIF 379 KB)Below is the link to the electronic supplementary material."} +{"text": "In the crystal, the mol\u00adecules are assembled into two-dimensional layers 33H28N2O4, comprises an indole unit (A), an iso\u00adquinoline moiety (B) and a benzene ring (C). The dihedral angles between these groups are A/B = 57.47\u2005(1), A/C = 18.48\u2005(1) and B/C = 57.97\u2005(1) \u00b0. The ethyl acrylate group at the 2-position is nearly co-planar with the indole unit [3.81\u2005(2)\u00b0], while that at the 7-position is distinctly non-coplanar [52.64\u2005(1)\u00b0]. Intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the indole unit and benzene ring help to establish the clip-shaped conformation of the mol\u00adecule. In the crystal, the mol\u00adecules are assembled into two-dimensional layers via C\u2014H\u22efO hydrogen bonds, \u03c0\u2013\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions. Hirshfeld surface analysis illustrates that the greatest contributions are from H\u22efH (63.2%), C\u22efH/H\u22efC (15.4%) and O\u22efH/H\u22efO (14.8%) contacts. The terminal C2H5 group of one of the ethyl acrylate side chains is disordered over two positions of equal occupancy.The mol\u00adecule of title compound, C The benzene ring is almost parallel to the indole unit and hence intra\u00admol\u00adecular \u03c0\u2013\u03c0 inter\u00adactions are observed within the layers to 1.7889 (blue) a.u. -diacrylate -8-phen\u00adyliso\u00adquinoline (0.20\u2005mmol), Pd(OPiv)2 , L quinoline; 11.3\u2005mg, 20\u2005mol%], CuO and Cu(OTf)2 and the tube was purged with O2 three times, followed by addition of ethyl acrylate (1.0\u2005mmol) and anhydrous DCE . The formed mixture was stirred at 353\u2005K under Ar for 24\u2005h as monitored by TLC. The solution was then cooled to room temperature, and the solvent was removed under vacuum. The crude product was purified by column chromatography on silica gel to afford the pure product (55% yield). The recrystallization of the title compound in methanol afforded yellow block-shaped crystals. The synthesis is shown in Fig.\u00a06To a 10\u2005mL Schlenk tube was added indole substrate 1-(1Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl). Atoms C32 and C33 were refined as disordered over two partially occupied positions of equal occupancy.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021007829/zn2008sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021007829/zn2008Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021007829/zn2008Isup3.cmlSupporting information file. DOI: 2100362CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-020-76496-2, published online 11 November 2020Correction to: Ca,L, SR Ca2+\u00a0content and Ca2+\u00a0transient measurements\u2019.This Article contains errors in the Results section under subheading \u2018The I2+\u00a0content and found that SR Ca2+\u00a0content was significantly increased in the trained group compared to controls (\u2212\u20091.84\u2009\u00b1\u20090.4 (pA*s)/pF vs \u2212\u20091.25\u2009\u00b1\u20090.5 (pA*s)/pF n\u2009=\u20098/5 and 8/4, respectively,\u00a0p\u2009<\u20090.05; Fig. 6b).\u201d\u201cWe analyzed the integral of caffeine-induced NCX currents as an indicator of the SR Cashould read:2+\u00a0content and found that SR Ca2+\u00a0content was significantly increased in the trained group compared to controls (\u2212\u20091.84\u2009\u00b1\u20090.4 (pA*s)/pF vs \u2212\u20091.25\u2009\u00b1\u20090.5 (pA*s)/pF n\u2009=\u20098/5 and 8/5, respectively,\u00a0p\u2009<\u20090.05; Fig. 6b).\u201d\u201cWe analyzed the integral of caffeine-induced NCX currents as an indicator of the SR CaSecondly, this Article contains an error in Figure\u00a06 where the experimental numbers of the control columns are incorrect (8/4) in panel (b). The correct Figure 6 appears below as Figure"} +{"text": "The asymmetric unit contains one-half of the formula unit of the title compound. The crystal structure is stabilized by inter\u00admolecular C\u2014H\u22efO, C\u2014H\u22efCl and C\u2014Cl\u22ef\u03c0 inter\u00adactions, and short inter\u00admolecular Cl\u22efO and Cl\u22efCl contacts, forming a three-dimensional network. 24H16Cl12N2O4, is generated by a crystallographic inversion centre at the midpoint of the central C\u2014C bond. A kink in the mol\u00adecule is defined by a torsion angle of \u2212169.86\u2005(15)\u00b0 about this central bond of the alkyl bridge. The pyrrolidine ring is essentially planar [max. deviation = 0.014\u2005(1)\u2005\u00c5]. The cyclo\u00adhexane ring has a boat conformation, while both cyclo\u00adpentane rings adopt an envelope conformation. In the crystal structure, mol\u00adecules are linked by inter\u00admolecular C\u2014H\u22efO, C\u2014H\u22efCl and C\u2014Cl\u22ef\u03c0 inter\u00adactions, and short inter\u00admolecular Cl\u22efO and Cl\u22efCl contacts, forming a three-dimensional network.The mol\u00adecule of the title compound, C N-heterocyclic compounds are of inter\u00adest in the fields of synthetic organic chemistry, coordination chemistry and medicinal chemistry because of their important biological properties -4,5,6,7,8,8-hexa\u00adchloro-2-{6-hex\u00adyl}-3a,4,7,7a-tetra\u00adhydro-1H-4,7-methano\u00adiso\u00adindole-1,3(2H)-dione, which provides multiple iner\u00admolecular non-covalent inter\u00adactions.QT = 0.9300\u2005(14)\u2005\u00c5, \u03b8 = 89.99\u2005(9)\u00b0, \u03c6 = 59.37\u2005(9)\u00b0], while both the cyclo\u00adpentane rings (C2\u2013C6 and C3\u2013C5/C8/C9) adopt an envelope conformation with the C4 atom bearing the di\u00adchloro\u00admethane group as the flap.The mol\u00adecule of the title compound is generated by a crystallographic inversion centre at the midpoint of the central C\u2014C bond. A kink in the mol\u00adecule is defined by the C10\u2014C11\u2013C12\u2014C12_a torsion angle of \u2212169.86\u2005(15)\u00b0 about this central bond of the alkyl bridge Fig.\u00a01. The pyrIn the crystal structure, mol\u00adecules are linked by inter\u00admolecular C\u2014H\u22efO, C\u2014H\u22efCl and C\u2014Cl\u22ef\u03c0 inter\u00adactions Table\u00a01, and shoCrystal Explorer 17.5 , Cl\u22efCl , O\u22efH/H\u22efO , Cl\u22efO/O\u22efCl and H\u22efH inter\u00adactions. The remaining other weak inter\u00adactions (contribution percentages) are Cl\u22efC/C\u22efCl (3.2%), Cl\u22efN/N\u22efCl (1.4%) and C\u22efH/H\u22efC (0.2%).In order to visualize the inter\u00admolecular inter\u00adactions Table\u00a02 in the cet al., 2016H-4,7-methano\u00adiso\u00adindole-1,3(2H)-dione -2-(perfluoro\u00adpyridin-4-yl)-3a,4,7,7a-tetra\u00adhydro-1H-4,7-methano\u00adiso\u00adindole-1,3(2H)-dione -2-[(perfluoro\u00adpyridin-4-yl)\u00adoxy]-3a,4,7,7a-tetra\u00adhydro-1H-4,7-methano\u00adiso\u00adindole-1,3(2H)-dione \u00b0 with the 3,4-di\u00admeth\u00adoxy\u00adphenyl ring, which are attached to each other by an extended N\u2014CHIn the crystal of QOVCAH, the cyclo\u00adhexene ring adopts a boat conformation, and the five-membered rings have envelope conformations with the bridging atom as the flap. Their mean planes are oriented at a dihedral angle of 86.51\u2005(7)\u00b0. The mol\u00adecular structure is stabilized by a short intra\u00admolecular C\u2014H\u22efO contact. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds, forming chains propagating along [100]. The chains are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming slabs parallel to (001).PA and B, in the asymmetric unit, and MOJGAW in the monoclinic space group P21/n with one mol\u00adecule per asymmetric unit. The synthesis of both compounds is conducted using endo starting materials, and the same configuration is observed in the resulting crystal structures. In MOJFUP, steric inter\u00adactions between the ortho-fluorine atoms and the carbonyl oxygen atoms prevents free rotation about the nitro\u00adgen\u2013ipso-carbon bond, which is evidenced by separate 19F NMR peaks in solution for the ortho-F atoms. In mol\u00adecule A, the 2,3,5,6-tetra\u00adfluoro\u00adpyridine plane is rotated by 58.05\u2005(5)\u00b0 relative to the pyrrolidine plane and the corresponding dihedral angle for mol\u00adecule B is 61.65\u2005(7)\u00b0. The addition of an oxygen atom between N and C in the bridge between the ring systems in MOJGAW alleviates this steric restriction and only one 19F NMR peak in solution is observed for the ortho-F atoms; even so, the dihedral angle between the 2,3,5,6-tetra\u00adfluoro\u00adpyridine and pyrrolidine planes in the crystal of MOJGAW of 84.01\u2005(5)\u00b0 is larger than that found in MOJFUP.The compound MOJFUP crystallizes in the triclinic space group The main directional inter\u00adactions in the crystal structures of MOJFUP and MOJGAW are of the type C\u2014H\u22efO, C\u2014H\u22efF, C\u2014O\u22ef\u03c0, and C\u2014F\u22ef\u03c0. In both compounds, weak hydrogen-bonding inter\u00adactions are observed for the hydrogen atom(s) \u03b1 to the carbonyl groups and the olefinic hydrogen atoms . A weak inter\u00adaction is also observed for a bridge hydrogen atom in MOJGAW, C\u2014H\u22efF. The packing is further aided by \u03c0-inter\u00adactions with the pyridine ring in MOJGAW.R,4R,7R,7aS)-4,5,6,7,8,8-hexa\u00adchloro-3a,4,7,7a-tetra\u00adhydro-4,7-methano\u00adisobenzo\u00adfuran-1,3-dione were added 0.12\u2005mL (1\u2005mmol) of hexane-1,6-di\u00adamine and 25\u2005mL of di\u00admethyl\u00adformamide, and the mixture was stirred for 6\u2005h at 373\u2005K. Then, the reaction mixture was cooled to room temperature and poured into cold water. The obtained precipitate was filtered off, washed with water, recrystallized from chloro\u00adform and dried under vacuum. Yellow powder, yield 92%, m.p 404\u2013405\u2005K (decomp.). Analysis calculated for C24H16Cl12N2O4 (Mr = 821.80): C 35.08, H 1.96, N 3.41%; found: C 35.03, H 2.00, N 3.35%. ESI\u2013MS: m/z: 822.9 [Mr\u00a0+\u00a0H]+. 1H NMR (300.130\u2005MHz) in acetone-d6, inter\u00adnal TMS, \u03b4 (ppm): 1.29\u20133.43 , 3.86 . 13C{1H} NMR . \u03b4: 25.8 (2CH2), 27.2 (2CH2), 39.3 (4C\u2013H), 52.0 (2CH2), 79.3 (4CCl), 104.4 (2CCl2), 130.9 (2ClC=CCl) and 170.2 (4C=O). Off-white prismatic crystals suitable for X-ray analysis were obtained by slow evaporation of a chloro\u00adform\u2013hexane mixture.To 741\u2005mg (2\u2005mmol) of (3aUiso(H) = 1.2Ueq(C). Two reflections (100 and 002), affected by the incident beam-stop, and owing to poor agreement between observed and calculated intensities, two outliers (136 and 118) were omitted in the final cycles of refinement. Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021006952/vm2251sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021006952/vm2251Isup2.hklStructure factors: contains datablock(s) I. DOI: 2094787CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of the free base of the ubiquitous neurotransmitter serotonin is reported for the first time. H-indol-5-ol], C10H12N2O, has one mol\u00adecule in the asymmetric unit. The conformation of the ethyl\u00adamino side chain is gauche\u2013gauche . In the crystal, the mol\u00adecules are linked into a three-dimensional network by N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds.The title compound, serotonin or 5-hy\u00addroxy\u00adtryptamine (5-HT) [systematic name: 3-(2-amino\u00adeth\u00adyl)-1 The ethyl\u00adamino arm itself turns back toward the indole ring with a C8\u2014C9\u2014C10\u2014N2 torsion angle of \u221261.9\u2005(2)\u00b0.Serotonin or 5-hy\u00addroxy\u00adtryptamine (5-HT) is an indolamine with a 5-hy\u00addroxy substitution. In the solid state, serotonin crystallizes with one mol\u00adecule in the asymmetric unit Fig.\u00a01 in the cA\u22efO1 hydrogen bonds. Half of the amine H atoms link to the hy\u00addroxy groups of nearby mol\u00adecules through N2\u2014H2B\u22efO1 hydrogen bonds. There are no observed \u03c0\u2013\u03c0 stacking inter\u00adactions. Fig.\u00a02In the crystal, the serotonin mol\u00adecules are linked by a series of hydrogen bonds that produce a three-dimensional network in the solid state. The hy\u00addroxy groups form hydrogen bonds to the amine N atoms on an adjacent serotonin mol\u00adecules forming O1\u2014H1\u22efN2 hydrogen bonds. The indole N atoms form hydrogen bonds to the hy\u00addroxy groups of adjacent serotonin mol\u00adecules through N1\u2014H110H13N2O+ cationic form. These include the creatine sulfate monohydrate .A, H2A and H2B were found from a difference-Fourier map and were refined isotropically, using DFIX restraints with an N\u2014H(indole) distance of 0.87\u2005(1)\u2005\u00c5, N\u2014H(amine) distances of 0.90\u2005(1)\u2005\u00c5, and an O\u2014H distance of 0.86\u2005(1)\u2005\u00c5. Isotropic displacement parameters were set to 1.2 Ueq of the parent nitro\u00adgen atoms and 1.5 Ueq of the parent oxygen atom. All other hydrogen atoms were placed in calculated positions with C\u2014H = 0.93\u2005\u00c5 (sp2) or 0.97\u2005\u00c5 (sp3). Isotropic displacement parameters were set to 1.2 Ueq of the parent carbon atoms. The absolute structure of the crystal chosen for data collection was indeterminate in the present refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022002559/hb8014sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989022002559/hb8014Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022002559/hb8014Isup3.cmlSupporting information file. DOI: 2156646CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-[2-(4-fluoro-3-phen\u00adoxy\u00adbenzo\u00adyl)hydrazinecarbo\u00adthio\u00adyl]benzamide and its 4-meth\u00adoxy derivative highlights the significance of strong and weak hydrogen bonds. The difference in the contributions of atom\u2013atom contacts obtained from Hirshfeld surface analysis and fingerprint plots helps in distinguishing the variations in the crystal packing of the two compounds.Crystal structure analysis of N-[2-(4-fluoro-3-phen\u00adoxy\u00adbenzo\u00adyl)hydrazinecarbo\u00adthio\u00adyl]benzamide, C21H16FN3O3S, crystallizes in the monoclinic centrosymmetric space group P21/c and its mol\u00adecular conformation is stabilized via an intra\u00admolecular N\u2014H\u22efO hydrogen bond. The corresponding para-meth\u00adoxy derivative, namely, N-[2-(4-fluoro-3-phen\u00adoxy\u00adbenzo\u00adyl)hydrazinecarbo\u00adthio\u00adyl]-4-meth\u00adoxy\u00adbenzamide, C22H18FN3O4S, crystallizes in the monoclinic centrosymmetric space group C2/c. The supra\u00admolecular network mainly comprises N\u2014H\u22efO, N\u2014H\u22efS and C\u2014H\u22efO hydrogen bonds, which contribute towards the formation of the crystal structures for the two mol\u00adecules. The different inter\u00admolecular inter\u00adactions have been further analysed using Hirshfeld surface analysis and fingerprint plots.The compound R\u2013(C=O)\u2013NH\u2013NH\u2013(C=S)\u2013R\u2032 and find application in the synthesis of five- and six-membered heterocyclic compounds hydrazinecarbo\u00adthio\u00adyl]benzamide (A1) and N-[2-(4-fluoro-3-phen\u00adoxy\u00adbenzo\u00adyl)hydrazinecarbo\u00adthio\u00adyl]-4-meth\u00adoxy\u00adbenzamide (A2) in the current study. The mol\u00adecular conformations have been studied with respect to the various flexible bonds and the occurrence of various inter\u00admolecular inter\u00adactions that contribute towards the stability of the mol\u00adecules in the crystalline lattice has been investigated in detail via an investigation of the crystal packing and qu\u00adanti\u00adtative insights from Hirshfeld surface analysis.Substituted thio\u00adsemicarbazides (TSCs) constitute an important class of organic compounds with the general formula P21/c space group and A2 crystallizes in the centrosymmetric monoclinic C2/c space group. The mol\u00adecular structure comprises one fluoro-substituted phen\u00adoxy\u00adbenzoyl ring, a rigid and planar (C=O)\u2014NH\u2014NH\u2014(C=S) moiety and a benzamide ring. The bond lengths and bond angles are in accordance with the magnitudes in the literature. The mol\u00adecular conformations of A1 , the N2\u22efO3 distance being 2.555\u2005(2) and 2.589\u2005(4)\u2005\u00c5 in A1 and A2, respectively. The mol\u00adecular structure possesses four conformational degrees of freedom due to the free rotation with respect to the N1\u2014N2, C7\u2014O1, O1\u2014C1 and C15\u2014C16 single bonds. The torsion angles C13\u2014N1\u2014N2\u2014C14, C8\u2014C7\u2014O1\u2014C1, C7\u2014O1\u2014C1\u2014C2 and N3\u2014C15\u2014C16\u2014C21 are 163.27\u2005(16)/-143.5\u2005(4)\u00b0, 97.3\u2005(2)/149.6\u2005(5)\u00b0, 167.18\u2005(18)/148.1\u2005(4)\u00b0 and \u2212160.26\u2005(15)/-174.7\u2005(3)\u00b0 in A1/A2, respectively.Compound A1 crystallizes in the centrosymmetric monoclinic A1 Fig.\u00a01 and A2 , [3.384\u2005(4)\u2005\u00c5, 174.9\u2005(1)\u00b0, \u2212x\u00a0+\u00a01, y\u00a0+\u00a01, \u2212z\u00a0+\u00a0In the crystal structure of A1, the mol\u00adecules are primarily assembled through the presence of N3\u2014H3s Table\u00a01, formingnt Fig.\u00a03. AdjacenA2 Fig.\u00a04 primarilA2 Fig.\u00a04, formingng Fig.\u00a04. Inter\u00admet al., 2016A search for the di\u00adbenzoyl\u00adthio\u00adsemicarbazide skeleton, Ph\u2013(C=O)\u2013NH\u2013NH\u2013(C=S)\u2013NH\u2013(C=O)\u2013Ph was carried out in the Cambridge Structural Database , and A2, Fig.\u00a05b) and 5(c), show the important hydrogen bonds. The red and blue spots correspond to inter\u00admolecular inter\u00adactions that are less or greater than the sum of the van der Waals radii. The fingerprint plots depict the individual contributions of the different inter\u00adactions. The fingerprint plots for A1/A2 = 1.2Ueq or 1.5Ueq(C-meth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021001900/dj2021sup1.cifCrystal structure: contains datablock(s) A1, A2. DOI: 10.1107/S2056989021001900/dj2021A1sup2.hklStructure factors: contains datablock(s) A1. DOI: 10.1107/S2056989021001900/dj2021A2sup3.hklStructure factors: contains datablock(s) A2. DOI: Click here for additional data file.10.1107/S2056989021001900/dj2021A1sup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989021001900/dj2021A2sup5.cmlSupporting information file. DOI: 2063226, 2063225CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Their physicochemical properties can be modulated by an appropriate choice of N\u2010substituents and counteranion. Tested against a panel of human cancer cell lines, the complexes were shown to possess promising antiproliferative activity and to circumvent multidrug resistance. Interestingly, most derivatives also retained a significant cytotoxic activity against human cancer 3D cell cultures. Among them, the complex with R=4\u2010C6H4OMe and R\u2019=Me emerged as the best performer of the series, being on average about six times more active against cancer cells than a noncancerous cell line, and displayed IC50 values comparable to those of cisplatin in 3D cell cultures. Mechanistic studies revealed the ability of the complexes to release carbon monoxide and to act as oxidative stress inducers in cancer cells.A straightforward two\u2010step procedure via single CO removal allows the conversion of commercial [Fe A class of cationic diiron complexes containing a variable bridging iminium ligand displays promising antiproliferative activity towards a panel of 2D and 3D cancer cell lines, and a selectivity not correlated to a different degree of uptake in cancerous and noncancerous cells. The mode of action appears to be mainly related to the interference with cell redox processes , initiated by intracellular CO release. Remarkably, 3D cell culture studies constitute an excellent base for more advanced in\u2005vivo cancer research, surpassing the traditional assays on monolayer cultures;To the best of our knowledge, the present 3D investigation is one of the first ones ever reported on iron compounds. The proposed diiron complexes combine within a cationic structure two Cp rings, three carbonyls and a variable bridging aminocarbyne ligand, the latter tuning the water solubility, the lipophilicity and the activity. Several experiments aimed to clarify the mechanism of action of the complexes will be discussed.1A\u2013H were prepared by the thermal substitution reaction of the corresponding isocyanides with a slight molar excess of [Fe2Cp2(CO)4] in acetonitrile.2\u20135]+ 2} core, connected to a further CO and an aminocarbyne group occupying the two bridging sites. Along this series of complexes, the C(4)\u2010N(1) distance does not significantly vary, indicating a substantial double\u2010bond character which accounts for the complementary description of the bridging ligand as iminium (see below).New compounds were fully characterized by elemental analysis, IR and multinuclear NMR spectroscopy. In addition, the structures of ACF3SO3, 2C]+ was calculated and compared to that of the parent compound cis\u20101C with a bridging cyclohexyl\u2010isocyanide ligand (CNCy). DFT\u2010optimized structures and relevant bonding parameters are reported in Table\u2005S1 and Figure\u2005S1. In 1C, the (\u03bc\u2010C)\u2212N Mayer bond order (b.o.)iPr instead of CNCy].2\u2009C]+. The nature of the nitrogen substituents has a negligible influence, and for instance the corresponding b.o. is 1.54 in the case of [2A]+, the related bond length being consistent with the crystallographic characterization of [2A]CF3SO3 + as an almost perfect aminocarbyne\u2010iminium hybrid structure } ligands in dimetal complexes is not clearly defined in the literature, therefore we performed a DFT study to shed light on this point. The representative structure of C+ was ca2Cp2Cl(CO)(\u03bc\u2010CO){\u03bc\u2010CNMe(Cy)}], 6 . Based on a comparison of the 1H NMR and IR spectra with data available in the literature for similar compounds,6 exists in CDCl3 solution as a nearly equimolar mixture of two isomers, maintaining the cis\u2010geometry of the Cp ligands and differing in the orientation of the N\u2010substituents with respect to the chloride (E/Z isomers). This feature suggests that the (\u03bc\u2010C)\u2212N bond in the neutral 6 still holds a substantial double bond character. The X\u2010ray structure of the E isomer was determined and is shown in Figure\u2005S2, although the low quality of the crystals prevents a detailed discussion of bonding parameters.Then, in order to clarify the possible effect of the net ionic charge of the complex, we also synthesized and characterized the novel neutral derivative +, with only a slight decrease of the (\u03bc\u2010C)\u2212N bond order (from 1.58 to 1.52). It is interesting to note that the bond length of the bridging carbonyl in 6 is very close to that in 1C (1.187 vs. 1.188\u2005\u00c5), highlighting that the isocyanide/CO and aminocarbyne/Cl combinations supply very similar electron densities for back\u2010donation to the diiron framework.In agreement with the NMR evidence, DFT outcomes indicate that 2Cl2 solution), the vibrations due to the terminal carbonyl ligands occur in the ranges 2018\u20132028 (anti\u2010symmetric stretching) and 1985\u20131996\u2005cm\u22121 (symmetric stretching), while the band related to the bridging carbonyl is found at 1833\u20131853\u2005cm\u22121. Moreover, the absorption attributed to the carbyne\u2010N bond falls in between 1530 to 1602\u2005cm\u22121, the higher values being associated to the N\u2010substituents with higher electron\u2010donor ability. The 1H and 13C NMR spectra display one set of resonances, ascribable to a single isomeric form featuring the Cp ligands in mutual cis position, as found in the X\u2010ray structures and confirmed by 1H NOE experiments in the case of [4G]+ . Two resonances for the Cp and terminal CO ligands are distinguishable in the unsymmetrical complexes, due to hampered rotation around the partially double C\u2212N bond (see above); accordingly, such resonances coincide in the complexes [2F]+ and [5H]+, displaying two identical N groups. In the 13C NMR spectra, the bridging {CN} unit is detected at low fields , as it is typical for bridging aminocarbynes.[2B]+ features a 31P NMR resonance at 17\u2005ppm for the phosphonato group, and the 13C signal for the carbyne appears as a doublet (3JCP ca. 6\u2005Hz).Salient spectroscopic features of all complexes are summarized in Table\u2005S2, and NMR and IR spectra are supplied in Figures\u2005S3\u2013S32. In the IR spectra (CH2C]CF3SO3 and [2C]Cl in chlorinated solvents are ascribable to the presence of ion pairs (or higher aggregates);1H NMR spectra in protic polar solvents (D2O and CD3OD), where ion pairs are practically absent.Small differences between the spectroscopic data of Cl. The octanol/water partition coefficients (log\u2009Pow) were measured by a UV/Vis method and fall in between 0 and \u22121, reflecting an amphiphilic or slightly hydrophilic character. An exception is represented by [2B]BF4, wherein the marked hydrophilicity imparted by the diethylphosphonato does not appear sufficiently balanced (log\u2009Pow<\u22121.5). Conversely, those complexes with aromatic groups are less soluble in water and display increased affinity for n\u2010octanol (log\u2009Pow\u22480). The diiron compounds are inert in aqueous solution at room temperature, and undergo a slow degradation at 37\u2009\u00b0C. Under such conditions, a variable amount of the starting material was detected by 1H NMR in D2O or D2O/CD3OD mixtures after 72\u2005h.2C]CF3SO3 solution) was separated and identified as \u03b3\u2010Fe2O3 by Raman analysis. In addition, headspace GC analyses performed on 5\u2009% MeOH aqueous solutions of the compounds maintained at 37\u2009\u00b0C allowed the identification and quantitation of the carbon monoxide released over a 48\u2005h period (Table\u2005S4). These facts indicate that the thermal degradation process in aqueous solution leads to the progressive disassembly of the organometallic scaffold.II ions was previously associated to their cytotoxicity,A detailed study on the behaviour of a selection of the diiron complexes in aqueous media was undertaken. Experimental procedures are provided in the Experimental Section and in the Supporting Information, with data collected in Tables\u20051H NMR. Notably, a considerable fraction of the starting material was detected at the end of the experiment, with a limited formation of an orange\u2010brown precipitate analogous to that described above.Next, the stability of the diiron compounds in a cell culture medium (DMEM) solution at 37\u2009\u00b0C, over a shorter time range (24\u2005h), was assessed by 2\u20133]+ was the methylated derivative, which was also checked by HPLC separation . The expected diiron counterparts, that is, 1A\u2013H, could not be detected possibly due to their water insolubility. The [lysozyme+Me+] to [lysozyme] ratios, calculated on extracted ion chromatograms after protein peak reconstruction, are reported in Table\u2005N\u2010methyl groups did not react with lysozyme. Methylation is a common post\u2010translational modification of protein/enzymes (methyltransferase enzymes) but it has also been observed as a result of the interaction of proteins with various electrophiles.To gain insights into the reactivity of the diiron compounds with possible biomolecular targets, we selected lysozyme, a small model protein often used to test interactions with metal compounds. Figure\u2005S1. The ex2A]+, [2C\u2013E]+, [3F\u2013G]+, [5H]+ and [2B]+, [4G]+ were tested for their cytotoxic potential by means of the MTT assay, as reported in the Experimental Section. The most convenient counter anion in terms of preparation and handling/storage was chosen in each case; in particular, [2C]CF3SO3 was included in the study, while [2C]Cl was excluded, being highly moisture sensitive and thus affecting the accuracy of the weighting operation. The in\u2010house human cancer cell line panel contains examples of ovarian (2008), colon (HCT\u201015), pancreatic (PSN\u20101), and breast (MCF\u20107) cancers as well as of melanoma (A375). Cisplatin was used as a reference and tested under the same experimental conditions. The cytotoxicity parameters, expressed in terms of IC50 obtained after 72\u2005h of exposure, are listed in Table\u20052B]BF4 and [3F]CF3SO3 did not impact cell viability , whereas the other compounds elicited IC50 values in the micromolar range. Despite all cytotoxic complexes showed an average in\u2005vitro antitumor potential lower than that of cisplatin, [2C]CF3SO3 and [2D]CF3SO3 proved to be the most effective derivatives, with average IC50 values of 17.4 and 16.6\u2005\u03bcM , and showed a similar pattern of response over the five tested cell lines. In particular, [2D]CF3SO3 exhibits a comparable activity with respect to cisplatin against HCT\u201015, PSN\u20101 and MCF\u20107 cells.The diiron complexes [[3F\u2013G]+, H+ . Overall, it appears that slight structural variations on the iminium group may have a notable impact on the activity of the cations, although not obvious: for instance, [2D]CF3SO3 and [2E]CF3SO3 differ from each other in the nature and the position of peripheral arene substituents and display almost identical log\u2009Pow values, notwithstanding the former is significantly more cytotoxic and selective than the latter.As one of the main drawbacks of chemotherapeutics is the toxic effect toward noncancerous cells, we measured the cytotoxicity of tested complexes against a noncancerous cell line (HEK293) and calculated the selectivity index . All complexes exhibited activity levels very similar on both sensitive (MCF\u20107) and multi\u2010drug resistant (MCF\u20107ADR) cell lines; interestingly, the resistance factor (RF) values Table\u2005 range fr2D]CF3SO3 (log\u2009Pow=\u22120.25) together with the less active/selective [2E]CF3SO3 (log\u2009Pow=\u22120.27) and [3G]CF3SO3 (log\u2009Pow=+0.29), to be assessed for their accumulation in human pancreatic PSN\u20101 cancer cells and in untransformed HEK293 cells. Thus, cells were treated for 24\u2005h with equimolar concentrations (50\u2005\u03bcM) of the tested compounds. The cellular iron levels were quantified by means of GF\u2010AAS analysis, and the results, expressed as \u03bcg Fe per 106 cells, are shown in Figure\u20053G]CF3SO3 was ineffective in entering human cells, while both [2D]CF3SO3 and [2E]CF3SO3 were able to induce cellular iron accumulation, and no significant differences in the ability to enter cells were evidenced for the two complexes. Considering the log\u2009Pow values, the obtained outcomes highlight that the hydrophilic complexes [2D]CF3SO3 and [2E]CF3SO3 are significantly more effective in entering cells compared to the more lipophilic one [3G]CF3SO3. In addition, both [2D]CF3SO3 and [2E]CF3SO3 led to major iron accumulation in untransformed cells with respect to pancreatic cancer cells. Intriguingly, these findings attest that their selectivity towards cancer cells with respect to untransformed ones cannot be attributable to their different ability to cross cellular plasmalemma and to enter human cells, and the reason of their preferential activity against cancer cells might be dependent on their ability to interfere with a cancer specific target.In order to correlate the cytotoxic potential to the ability of the newly developed complexes to enter cells, we performed uptake experiments. To this purpose, we selected the most promising compound CF3SO3 and [3F]CF3SO3 were ineffective against cancer cells spheroids, whereas [2D]CF3SO3 was the most effective complex among the series, thus indicating its ability to better penetrate the spheroid and reach the hypoxic core. It is noteworthy that [2D]CF3SO3 exhibited a level of activity either comparable (PSN\u20101 cells) or even superior (2008 cells) to that of cisplatin.The remarkable cell\u2010killing effect observed in 2D cultured cells prompted us to evaluate the in\u2005vitro antitumor activity of the complexes on 3D cell cultures. Differently from 2D monolayer culture, 3D spheroid cell culture systems comprise cancer cells in various cell growth stages. Consequently, the multicellular cancer spheroid model is recognized to better reflect the tumour mass in\u2005vivo regarding drug permeation, cell interactions, gene expression, hypoxia and nutrient gradients with respect to monolayer cell cultures, making 3D models more predictive than conventional 2D monolayer cultures in screening antitumor drugs.50 ratio related to 3D and 2D experiments ranging from 2 to 5 for both cell lines; in particular, this ratio is approximately 2 for the cytotoxic diiron complexes against the 2008\u2005cell line, while the corresponding value for cisplatin is 26. Note that a substantial drop of antiproliferative activity has been often recognized for potential anticancer metal drugs in 3D cell cultures compared to the corresponding 2D ones, highlighting the need for a combination of both models for screening of compounds.An important aspect of the collected data is that only a limited decrease of activity is observed with the cytotoxic compounds, the IC2C]CF3SO3 and [2D]CF3SO3 to release carbon monoxide inside PSN\u20101 pancreatic cancer cells and HEK293 noncancer cells. A fluorometric test was conducted over a short time interval (30\u2005min) by means of a CO sensing dye (L)(\u03bc\u2010CO){\u03bc\u2010CNR(Me)}]0/+.CF3SO3 and [2D]CF3SO3 determined a substantial increase in cellular ROS production, in a time\u2010dependent manner oxides has been recognized from water solution, see above). Otherwise, based on previous voltammetric measurements on [2E\u2013F]CF3SO3,2\u20133]+, some contribution to ROS might be provided by the partial generation of the neutral products 1A\u2013H upon CH3+ elimination (see above); in principle, the absence of a net positive charge in 1A\u2013H might favour the oxidation of these complexes or their possible derivatives in the cells.Iron is a redox active metal that can generate ROS in cells via the Fenton reaction, thus inducing cellular damage and ultimately leading to cell death.r Figure\u2005A. Remark3 Figure\u2005B. The ti2C]CF3SO3 and [2D]CF3SO3 to target TrxR both in cell\u2010free experiments and in cells. The two complexes showed a similar pattern of response, being completely ineffective in hampering TrxR in cell\u2010free experiment (data not shown) but elicited a substantial inhibition of the selenoenzyme in human pancreatic PSN\u20101 cancer cells . On the other hand, TrxR inhibition might also be consistent with the capacity of the complexes to methylate specific substrates, as we have assessed for lysozyme. Actually, it was previously described that several alkylating agents are able to selectively modify the redox active Cys or Sec of TrxR, thus causing irreversible inhibition.It has recently emerged that the redox stress induced by some iron complexes might also be attributed to their ability to inhibit the selenoenzyme thioredoxin reductase (TrxR). In particular, Rigobello and co\u2010workers reported the capacity of a ferrocenyl diphenol and of a Tamoxifen\u2010like ferrocifen to target and hamper TrxR activity.s Figure\u2005. In part2Cp2(CO)4]. They have ideal properties for a potential anticancer drug, such as the presence of a nontoxic metal element, appreciable water solubility and amphiphilicity, and stability in aqueous media.2D]CF3SO3, with a 4\u2010methoxy phenyl substituent on the bridging hydrocarbyl ligand, was revealed to be the most promising and exhibited a noteworthy selectivity toward cancer cell lines compared to noncancerous cells. According to cellular\u2010uptake experiments, such selectivity is not correlated to a different ability to enter cancerous or noncancerous cells. Remarkably, the cytotoxicity profile of the complexes does not substantially decrease in 3D cell cultures, in contrast to what has often been observed in the literature upon comparison of 2D/3D studies on various transition metal compounds. Interestingly, the activity exhibited by [2D]CF3SO3 in the ovarian cancer cell 3D model was even superior to that of cisplatin. Experiments on selected complexes outlined their power to behave as carbon monoxide\u2010releasing molecules (CORMs) inside cells, to unbalance cellular redox homeostasis and to alkylate biological targets. The preferential CO release in cancer cells with respect to untransformed cells could, at least in part, be consistent with the ability to target TrxR. Actually, it is well known that the Trx system is overexpressed in cancer cells, and has been recognized as a tumour\u2010specific target for the development of selective anticancer agents. In summary, we suggest that a simple diiron platform might offer an arsenal of tools not available for widely investigated ferrocenes, paving the way for the development of iron\u2010based drugs with optimized features. Further biochemical and biophysical studies will be performed to clarify in more detail the mechanism of action of this new class of organometallic anticancer candidates.We have described a synthetic strategy to obtain a family of diiron carbonyl complexes containing a bridging aminocarbyne/iminium hybrid ligand from the readily available [FeGeneral experimental details: [Fe2Cp2(CO)4] (99\u2009%) was purchased from Strem Chemicals, other reactants and solvents were obtained from Alfa Aesar, Merck, Apollo Scientific or TCI Chemicals and were of the highest purity available. Isocyanides were stored at 4\u2009\u00b0C or \u221220\u2009\u00b0C and used as received; contaminated labware was treated with bleach. Methyl triflate was stored under N2 at 4\u2009\u00b0C; allyl iodide and triethyloxonium tetrafluoroborate (1\u2005M solution in CH2Cl2) were stored under N2 at \u221220\u2009\u00b0C. Compounds [1A\u2013H]2D\u2013F]CF3SO3X , [2C]Cl, [5H]CF3SO3 (hygroscopic) and 6 were stored under N2; all other Fe compounds being air\u2010 and moisture\u2010stable in the solid state. NMR spectra were recorded at 25\u2009\u00b0C on a Bruker Avance II DRX400 instrument equipped with a BBFO broadband probe. Chemical shifts (expressed in parts per million) are referenced to the residual solvent peaks1H, 13C) or to external standards46 . In [D6]DMSO/D2O mixtures, 1H chemical shifts are referenced to the [D5]DMSO signal as in pure [D6]DMSO (\u03b4/ppm=2.50); in D2O/CD3OD mixtures, 1H chemical shifts are referenced to the CD3OD residual peak as CH3OH in pure D2O (\u03b4/ppm=3.34). 1H and 13C spectra were assigned with the assistance of 1H{31P}, 13C DEPT 135, 1H,1H COSY, 1H\u201013C gs\u2010HSQC and 1H\u201013C gs\u2010HMBC experiments.3 stored in the dark over Na2CO3 was used for NMR analysis. IR spectra of solid samples (650\u20134000\u2005cm\u22121) were recorded on a Perkin Elmer Spectrum One FTIR spectrometer, equipped with a UATR sampling accessory; IR spectra of solutions were recorded using a CaF2 liquid transmission cell (2300\u20131500\u2005cm\u22121) on a Perkin Elmer Spectrum 100 FTIR spectrometer. UV/Vis spectra (250\u2013800\u2005nm) were recorded on a Ultraspec 2100 Pro spectrophotometer using PMMA cuvettes (1\u2005cm path length). IR and UV/Vis spectra were processed with Spectragryph software.2Cp2(CO)3(CNR)], [1A[Fe\u2013H].General procedure. In a 250\u2005mL round\u2010bottom flask equipped with a reflux condenser under N2, the selected isocyanide, or its solution in MeCN , was added dropwise to a suspension of [Fe2Cp2(CO)4] (1.6\u2005equiv) in anhydrous MeCN (80\u2013100\u2005mL). Alkyl isocyanides: the mixture was heated at reflux (T\u2265100\u2009\u00b0C) for 8\u2005h, then stirred at room temperature for additional 14\u2005h. Aryl isocyanides: the mixture was stirred at room temperature for 14\u2005h then heated at reflux (T\u2265100\u2009\u00b0C) for 1\u20133\u2005h. Next, conversion was checked by IR and the dark red\u2010brown suspension was dried under vacuum (40\u2009\u00b0C). The resulting solid, consisting of a mixture of [Fe2Cp2(CO)4] and [Fe2Cp2(CO)3(CNR)], was directly used in the following alkylation procedure without any purification.2Cp2(CO)2(\u03bc\u2010CO){\u03bc\u2010CNMe(1H\u2010indol\u20106\u2010yl)}]CF3SO3, [2A]CF3SO3CF3SO3 is soluble in MeCN, THF, MeOH, CH2Cl2, less soluble in CH2Cl2, CHCl3, insoluble in Et2O, toluene and water. X\u2010ray quality crystals of [2A]CF3SO3 were obtained from an acetone solution layered with Et2O and settled aside at \u221220\u2009\u00b0C. Anal. calcd. for C25H21F3Fe2N2O6S: C 46.47, H 3.28, N 4.34; found: C 46.09, H 3.28, N 4.34. IR (solid state): \u03bd\u02dc/cm\u22121=3330w\u2010br (\u03bdNH), 3114w, 2004\u2005s (\u03bdCO), 1985\u2005s (\u03bdCO), 1833\u2005s (\u03bd\u03bc\u2010CO), 1583w\u2010sh, 1555\u2005m, 1540\u2005m (\u03bdCN), 1475w, 1458w, 1433w, 1420w, 1397\u2005m, 1361w, 1345w, 1323w, 1290\u20131276\u2005m, 1244\u2005s (\u03bdSO3), 1223\u2005s (\u03bdSO3), 1154\u2005s (\u03bdSO3), 1107\u2005m\u2010sh, 1067w, 1028\u2005s, 1016\u2005s\u2010sh, 1002\u2005m\u2010sh, 942w, 931w, 893w, 875w, 860\u2005m, 845\u2005m, 817w, 797\u2005m, 772\u2005s, 756w\u2010sh, 743\u2005m\u2010sh, 728\u2005m. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2022\u2005s (\u03bdCO), 1992\u2005m\u2010sh (\u03bdCO), 1836\u2005s (\u03bd\u03bc\u2010CO), 1554\u2005m, 1542\u2005m (\u03bdCN). IR (MeCN): \u03bd\u02dc/cm\u22121=2023\u2005s (\u03bdCO), 1991\u2005m\u2010sh (\u03bdCO), 1836\u2005s (\u03bd\u03bc\u2010CO), 1554\u2005m, 1542\u2005m\u2010sh (\u03bdCN). 1H NMR (CD3CN): \u03b4/ppm=9.78 , 7.85 , 7.70 , 7.48 , 7.37 , 6.67\u22126.62 ; 5.39, 4.65 , 4.53 . 13C{1H} NMR (CD3CN): \u03b4/ppm=324.9 (CN), 255.9 (\u03bc\u2010CO); 209.8, 209.2 (CO); 144.6 (C2), 136.3 (C5), 129.1 (C8), 128.8 (C6), 118.8 (C3), 117.3 (C9), 113.8 (C4), 103.5 (C7); 91.2, 91.0 (Cp); 58.5 (C1). 19F{1H} NMR (CD3CN): \u03b4/ppm=\u221279.3. 1H NMR (CDCl3): \u03b4/ppm=9.57 , 8.0\u20137.0 , , 7.42\u20137.39 , 6.60 ; 5.41, 4.63 ; 4.56 .In a 150\u2005mL Schlenk tube under N3SO3[2B]CF. In a 150\u2005mL Schlenk tube under N2, CF3SO3Me was added dropwise to a dark red solution of 1B (1.38\u2005mmol) in anhydrous CH2Cl2 (40\u2005mL) under stirring. Therefore, the mixture was stirred at room temperature for 7\u2005h and conversion was checked by IR (CH2Cl2). Next, the solution was moved on top of an alumina column . Impurities were eluted with neat CH2Cl2, THF and MeCN, then a red band containing the title product was eluted with MeCN/MeOH . Volatiles were removed under vacuum (40\u2009\u00b0C), affording a red, highly hygroscopic solid. Yield: quantitative. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2028\u2005s (\u03bdCO), 1997w\u2010sh (\u03bdCO), 1836\u2005m (\u03bd\u03bc\u2010CO). 1H NMR (CDCl3): \u03b4/ppm=5.32 , 5.23 , 4.93 , 4.38 , 4.23\u20134.10 , 1.30 . 19F{1H} NMR (CDCl3): \u03b4/ppm=\u221278.2. 31P{1H} NMR (CDCl3): \u03b4/ppm=17.3.4[2B]BF. In a 500\u2005mL round bottom flask, [2B]CF3SO3 was suspended in water (100\u2005mL) with vigorous stirring until completely dissolved then treated with NaBF4 (500\u2005mg). The red solution was extracted with CH2Cl2 (3\u00d720\u2005mL) and the combined organic fractions were dried under vacuum. The residue was suspended in water and the ion exchange procedure with NaBF4 was repeated (\u00d73); 19F{1H} NMR analysis of the final CH2Cl2 solution indicated the complete removal of CF3SO3\u2212 anions. Therefore, volatiles were removed under vacuum; the residue was dissolved in CH2Cl2 and filtered over celite. A red foamy hygroscopic solid, obtained upon volatiles removal without heating, was dried under vacuum and stored under N2. Yield: 751\u2005mg, 90\u2009% (with respect to 1B). Compound [2B]BF4 is soluble in MeOH, MeCN, CH2Cl2, CHCl3, water, insoluble in Et2O and hexane. Anal. calcd. for C20H25BF4Fe2NO6P: C 39.71, H 4.17, N 2.31; found: C 39.20, H 4.25, N 2.26. IR (solid state): \u03bd\u02dc/cm\u22121=3117w, 2987w, 2938w, 2913w, 2875w, 2012\u2005s (\u03bdCO), 1986\u2005s\u2010sh (\u03bdCO), 1823\u2005s (\u03bd\u03bc\u2010CO), 1568\u2005m (\u03bdCN), 1479w, 1446w, 1435w, 1421w, 1399\u2005m, 1370w\u2010sh, 1297\u20131286w, 1254\u2005m, 1225\u2005m\u2010sh, 1180w, 1163w, 1075\u2005m\u2010sh, 1033\u2005s\u2010sh (\u03bdBF4), 1008\u2005s, 973\u2005s, 951\u2005s\u2010sh, 904\u2005m, 864\u2005m\u2010sh, 851\u2005m, 837\u2005m\u2010sh, 780s, 745\u2005s, 710\u2005m\u2010sh. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2028\u2005s (\u03bdCO), 1996w (\u03bdCO), 1836\u2005m (\u03bd\u03bc\u2010CO), 1574w (\u03bdCN). IR (MeCN): \u03bd\u02dc/cm\u22121=2026\u2005s (\u03bdCO), 1994w (\u03bdCO), 1838\u2005m (\u03bd\u03bc\u2010CO), 1569w (\u03bdCN). 1H NMR (CDCl3): \u03b4/ppm=5.29 , 5.15 , 4.93 , 4.35 , 4.24\u20134.10 , 1.30 . No changes were observed in the 1H spectrum after 14\u2005h at room temperature. 13C{1H} NMR (CDCl3): \u03b4/ppm=323.7 , 254.7 (\u03bc\u2010CO); 207.4, 207.1 (CO); 90.3, 90.1 (Cp); 63.3 (C3), 62.5 , 53.2 (C1), 16.4 . 19F{1H} NMR (CDCl3): \u03b4/ppm=\u2010151.9 (10BF4), \u2212152.0 (11BF4). 31P{1H} NMR (CDCl3): \u03b4/ppm=17.3.3SO3[2C]CF. In a 500\u2005mL round bottom Schlenk flask under N2, CF3SO3Me was added dropwise to a dark red suspension of 1C (22.5\u2005mmol) in anhydrous CH2Cl2 (100\u2005mL) under stirring. Therefore, the mixture was stirred at room temperature for 4\u2005h and the conversion was checked by IR (CH2Cl2). Next, the dark red solution was moved on top of an alumina column . Impurities were eluted with neat CH2Cl2 and THF, then a red band containing [2C]CF3SO3 was eluted with MeCN/MeOH . A red band, containing a minor fraction of [2C]Cl, was collected using MeOH as eluent. Volatiles were removed under vacuum from the MeCN/MeOH solution and residue was triturated in Et2O. The suspension was stirred at room temperature for a few h then filtered. The resulting red solid was washed with toluene, Et2O and dried under vacuum (40\u2009\u00b0C). Yield: 11.44\u2005g, 85\u2009%. Compound [2C]CF3SO3 is soluble in MeCN, MeOH, CH2Cl2, CHCl3, less soluble in water, poorly soluble in toluene, insoluble in Et2O, hexane. X\u2010ray\u2010quality crystals of [2C]CF3SO3 were obtained from a CH2Cl2 solution layered with heptane and settled aside at \u221220\u2009\u00b0C. Anal. calcd. for C22H24F3Fe2NO6S: C 44.10, H 4.04, N, 2.34; found: C 44.47, H 4.04, N, 2.34. IR (solid state): \u03bd\u02dc/cm\u22121=3102w, 2937w, 2862w, 2185w, 2146w, 2006\u2005s (\u03bdCO), 1982\u2005s\u2010sh (\u03bdCO), 1822\u2005s (\u03bd\u03bc\u2010CO), 1565\u2005m (\u03bdCN), 1544\u2005m\u2010sh, 1452w, 1435w, 1421w, 1402w, 1366w, 1352w, 1321w, 1272\u2005s\u2010sh, 1257\u2005s (\u03bdSO3), 1223\u2005s\u2010sh (\u03bdSO3), 1150s (\u03bdSO3), 1056\u2005m, 1029\u2005s, 990\u2005m, 864\u2005m, 854\u2005m, 796\u2005s, 746\u2005s. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2020s (\u03bdCO), 1988w\u2010sh (\u03bdCO), 1835\u2005m (\u03bd\u03bc\u2010CO), 1567w (\u03bdCN). IR (MeCN): \u03bd\u02dc/cm\u22121=2021\u2005s (\u03bdCO), 1988w\u2010sh (\u03bdCO), 1835\u2005m (\u03bd\u03bc\u2010CO), 1568w (\u03bdCN). 1H NMR (CDCl3): \u03b4/ppm=5.36, 5.27 ; 4.68 , 4.06 , 2.81 , 2.15 , 2.03\u20131.94 , 1.86\u20131.72 , 1.53\u20131.35 , 1.34\u20131.20 . No changes were observed in the 1H spectrum after 14\u2005h at room temperature. 13C{1H} NMR (CDCl3): \u03b4/ppm=316.4 (CN), 255.3 (\u03bc\u2010CO); 208.5, 207.6 (CO); 90.3, 90.1 (Cp); 79.8 (C2), 46.8 (C1), 31.2 (C5), 30.7 (C4), 26.1 (C3), 26.0 (C6), 25.1 (C7). 19F{1H} NMR (CDCl3): \u03b4/ppm=\u221278.2. 1H NMR (CD3OD): \u03b4/ppm=5.40, 5.36 , 4.07 , 2.41 , 2.17\u20132.07, 2.03\u20131.91, 1.85\u20131.50, 1.42\u20131.32 .[2C]Cl. Compound [2C]CF3SO3 was dissolved in MeOH (2\u2005mL) then moved on top of an Amberlyst 15 column . NaCF3SO3 was eluted with neat MeOH then a red band containing [2C]+ was eluted with NaCl\u2010saturated MeOH. Volatiles were removed under vacuum and the residue was suspended in MeCN. The suspension was filtered over celite; the filtrate was checked for absence of CF3SO3\u2212 by 19F NMR then dried under vacuum (40\u2009\u00b0C). The resulting red, hygroscopic solid was stored under N2. Yield: 83\u2005mg, 93\u2009%. Compound [2C]Cl shows the same solubility pattern as the triflate salt. X\u2010ray quality crystals of [2C]Cl\u2009\u22c5\u20092.34H2O were obtained from a CH2Cl2 solution layered with hexane and settled aside at \u221220\u2009\u00b0C. Anal. calcd. for C21H24ClFe2NO3: C 51.94, H 4.98, N 2.88; found: C 51.99, H 5.02, N 2.83. IR (solid state): \u03bd\u02dc/cm\u22121=3376\u2005m\u2010br* (\u03bdOH), 3098\u20133059w, 2933\u2005m, 2858w, 2180\u20132160w, 1997\u2005s (\u03bdCO), 1969\u2005s\u2010sh (\u03bdCO), 1811\u2005s (\u03bd\u03bc\u2010CO), 1662w, 1631w, 1565\u2005s (\u03bdCN), 1543\u2005m, 1450\u2005m, 1434w, 1419\u2005m, 1400\u2005m, 1350w, 1319w, 1263w\u2010sh, 1249w, 1173w, 1154w, 1054\u2005m, 1026w, 1015w, 990\u2005m, 924w\u2010sh, 893w\u2010sh, 852\u2005m, 795\u2005s, 746\u2005s, 729\u2005s\u2010sh. *Due to moisture. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2018\u2005s (\u03bdCO), 1985w\u2010sh (\u03bdCO), 1833\u2005m (\u03bd\u03bc\u2010CO), 1569w (\u03bdCN). IR (MeCN): \u03bd\u02dc/cm\u22121=2020s (\u03bdCO), 1987w\u2010sh (\u03bdCO), 1834\u2005m (\u03bd\u03bc\u2010CO), 1568w (\u03bdCN). 1H NMR (CDCl3): \u03b4/ppm=5.51, 5.39 ; 4.75\u20134.66 , 4.14 , 3.16 , 2.15 , 2.03\u20131.93 , 1.84\u20131.72 , 1.51\u20131.35 , 1.33\u20131.23 . No changes were observed in the 1H spectrum after 24\u2005h at room temperature. 13C{1H} NMR (CDCl3): \u03b4/ppm=316.2 (CN), 255.8 (\u03bc\u2010CO); 208.7, 207.8 (CO); 90.7, 90.3 (Cp); 79.7 (C2), 47.1 (C1), 31.3 (C5), 31.0 (C4), 26.1 (C3), 26.0 (C6), 25.2 (C7). 35Cl NMR (CDCl3): \u03b4/ppm=7.61 (\u0394\u03bd1/2=266\u2005Hz).2, CF3SO3Me was added dropwise to a dark red solution of 1B (4.63\u2005mmol) in anhydrous CH2Cl2 (40\u2005mL) under stirring. The mixture was stirred at room temperature for 5.5\u2005h then moved on top of an alumina column . Impurities were eluted with CH2Cl2 and CH2Cl2/THF , then a red band containing [2D]CF3SO3 was eluted with neat MeCN. Volatiles were removed under vacuum (40\u2009\u00b0C) to afford a red foamy solid. Yield: 2.446\u2005g, 85\u2009%. Compound [2D]CF3SO3 is soluble in MeCN, MeOH, CH2Cl2, CHCl3, water, insoluble in Et2O and hexane. X\u2010ray quality crystals of [2D]CF3SO3 were obtained from a CHCl3 solution layered with hexane and settled aside at \u221220\u2009\u00b0C. Anal. calcd. for C23H20F3Fe2NO7S: C 44.33, H 3.24, N 2.25; found: C 43.92, H 3.31, N 2.17. IR (solid state): \u03bd\u02dc/cm\u22121=3113w, 3091w, 2946w, 2849w, 2011\u2005s (\u03bdCO), 1978\u2005s (\u03bdCO), 1821\u2005s (\u03bd\u03bc\u2010CO), 1608w, 1563\u2005m, 1539\u2005m (\u03bdCN), 1506\u2005s, 1468w, 1449w, 1433w, 1420w, 1395\u2005m, 1304w\u2010sh, 1278\u2005s, 1251\u2005s (\u03bdSO3), 1224\u2005s\u2010sh (\u03bdSO3), 1176\u2005m\u2010sh, 1146\u2005s (\u03bdSO3), 1117\u2005m\u2010sh, 1108\u2005m, 1067w, 1028\u2005s, 890w, 855\u2005s, 825w\u2010sh, 773\u2005s, 756\u2005m\u2010sh, 736w, 702\u2005s. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2021\u2005s (\u03bdCO), 1989w (\u03bdCO), 1836\u2005m (\u03bd\u03bc\u2010CO), 1607w, 1562w, 1540w (\u03bdCN), 1507\u2005m. IR (MeCN): \u03bd\u02dc/cm\u22121=2024\u2005s (\u03bdCO), 1991w (\u03bdCO), 1837\u2005m (\u03bd\u03bc\u2010CO), 1610w, 1563w, 1539w (\u03bdCN), 1508w. 1H NMR (CDCl3): \u03b4/ppm=8.3\u20137.6 , 7.10 ; 5.43, 4.78 ; 4.54 , 3.90 . 19F{1H} NMR (CDCl3): \u03b4/ppm=\u221278.1.The title compound was prepared with minor modifications of the literature procedure.[3F]I. The preparation of the title compound was optimized with respect to the literature .2, allyl iodide was added dropwise to a dark red solution of 1F (1.54\u2005mmol) in anhydrous MeCN (30\u2005mL) under vigorous stirring. The mixture was stirred for 4.5\u2005h under protection from the light and conversion was checked by IR (MeCN). Next, the dark red solution was moved on top of an alumina column . Impurities were eluted with neat Et2O, THF and MeCN, then a red band containing the title product was eluted with MeCN/MeOH . Volatiles were removed under vacuum (40\u2009\u00b0C) and the residue was triturated in Et2O. The suspension was stirred at room temperature then filtered. The resulting red solid was washed with Et2O and dried under vacuum (40\u2009\u00b0C). Yield: 646\u2005mg, 78\u2009%. Compound [3F]I is soluble in MeOH, CH2Cl2, CHCl3, poorly soluble in water, insoluble in Et2O, hexane. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2021\u2005s (\u03bdCO), 1989w (\u03bdCO), 1835\u2005m (\u03bd\u03bc\u2010CO), 1585w (\u03bdCN). 1H NMR (CDCl3): \u03b4/ppm=6.09\u20135.97 , 5.53\u20135.46 ; 5.45, 5.53 ; 5.17\u20135.11 , 4.25 .3SO3[3F]CF. In a 50\u2005mL round bottom flask, a solution of Ag(CF3SO3) in MeCN (5\u2005mL) was added to a dark red suspension of [3F]I in MeCN (6\u2005mL) at 0\u2009\u00b0C. The mixture stirred at 0\u2009\u00b0C for 2.5\u2005h while maintaining the system under protection from the light. The resulting suspension (dark red solution+yellow AgI precipitate) was filtered over celite. Volatiles were removed under vacuum from the filtrate solution and the residue was triturated in a Et2O/toluene mixture. The suspension was stirred at room temperature for a few h, then filtered. The resulting red solid was washed with toluene, Et2O and dried under vacuum (40\u2009\u00b0C). Yield: 928\u2005mg, 94\u2009%. Compound [3F]CF3SO3 is soluble in MeCN, acetone, MeOH, water, less soluble in CH2Cl2 > CHCl3, poorly soluble in toluene, insoluble in Et2O. Anal. calcd. for C19H18F3Fe2NO6S: C 40.96, H, 3.26, N, 2.51; found: C 41.20, H, 3.20, N, 2.72. IR (solid state): \u03bd\u02dc/cm\u22121=3106w, 2213w, 2178w, 2012\u2005s (\u03bdCO), 1988\u2005s (\u03bdCO), 1813\u2005s (\u03bd\u03bc\u2010CO), 1644w, 1581\u2005m (\u03bdCN), 1435w, 1416w, 1395w, 1261\u2005s (\u03bdSO3), 1243\u2005s\u2010sh, 1222\u2005m\u2010sh (\u03bdSO3), 1169\u2005s (\u03bdSO3), 1050\u2005m, 1028\u2005s, 989w\u2010sh, 931\u2005m, 888\u2005m, 854\u2005m, 758\u2005s, 666\u2005m. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2022\u2005s (\u03bdCO), 1990w (\u03bdCO), 1836\u2005m (\u03bd\u03bc\u2010CO), 1584\u2005m (\u03bdCN). IR (MeCN): \u03bd\u02dc/cm\u22121=2023\u2005s (\u03bdCO), 1990w (\u03bdCO), 1836\u2005m (\u03bd\u03bc\u2010CO), 1582\u2005m (\u03bdCN). 1H NMR (CDCl3): \u03b4/ppm=5.98 , 5.50\u20135.45 ; 5.34, 5.32 ; 5.30\u20135.23 , 5.15\u20135.08 , 4.17 . No changes were observed in the 1H spectrum after 14\u2005h at room temperature. 13C{1H} NMR (CDCl3): \u03b4/ppm=318.1 (CN), 255.3 (\u03bc\u2010CO); 208.1, 207.8 (CO), 130.2 (C3), 122.2 (C4), 121.0 ; 90.20, 90.16 (Cp); 70.6 (C2), 51.3 (C1). 19F{1H} NMR (CDCl3): \u03b4/ppm=\u221278.2. 1H NMR ([D6]acetone): \u03b4/ppm=6.33\u20136.19 , 5.62 ; 5.58, 5.54 ; 5.41\u20135.16 , 4.28 .2, methyl iodide was added to a dark red solution of 1G (0.48\u2005mmol) in anhydrous MeCN (8\u2005mL). The mixture was stirred for 2.5\u2005h and conversion was checked by IR (MeCN). Next, the dark red solution was moved on top of an alumina column . Impurities were eluted with neat CH2Cl2 and THF, then a red band containing the title product was eluted with THF/MeOH . Next, volatiles were removed under vacuum and the residue was dissolved in CH2Cl2. The suspension was filtered over celite and the filtrate was taken to dryness under vacuum (40\u2009\u00b0C), affording a dark red solid. Yield: 208\u2005mg, 69\u2009%. A considerable decrease in yield and purity was observed for reactions carried out in refluxing CHCl3 or in MeCN at lower temperatures. Anal. calcd. for C25H20Fe2INO3: C 48.35, H 3.25, N 2.56; found: C 48.20, H 3.15, N 2.49. Compound [3G]I is soluble in MeOH, CH2Cl2, CHCl3, sparingly soluble in water, insoluble in hexane. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2021\u2005s (\u03bdCO), 1989\u2005m\u2010sh (\u03bdCO), 1836\u2005m (\u03bd\u03bc\u2010CO), 1564w, 1545w, 1538w (\u03bdCN). 1H NMR (CDCl3): \u03b4/ppm=8.10 , 7.98\u22127.95 , 7.94\u22127.77 , 7.66\u22127.52 , 7.55\u22127.47 (C10H7); 5.62, 4.82 ; 4.75 .In a 25\u2005mL Schlenk tube under N2, [Et3O]BF4 , was added dropwise to a dark red solution of 1G (0.724\u2005mmol) in anhydrous MeCN (30\u2005mL) at 0\u2009\u00b0C under vigorous stirring. Therefore, the mixture was stirred for 3\u2005h, while allowing to reach room temperature. Next, conversion was checked by IR (MeCN) then volatiles were removed under vacuum. The residue was dissolved in a small volume of THF then moved on top of an alumina column . Impurities were eluted with neat Et2O and THF, then a red band containing the title product was eluted with MeCN/MeOH . Volatiles were removed under vacuum (40\u2009\u00b0C) and the residue was triturated in Et2O. The suspension was stirred at room temperature for a few h then filtered. The resulting dark red solid was washed with Et2O and dried under vacuum (40\u2009\u00b0C). Yield: 284\u2005mg, 66\u2009%. A considerable decrease in yield and purity was observed for reactions carried out in CH2Cl2 or in MeCN at room temperature. Compound [4G]BF4 is soluble in MeOH, MeCN, CH2Cl2, CHCl3, poorly soluble in Et2O, insoluble in water. Anal. calcd. for C26H22BF4Fe2NO3: C 52.49, H 3.72, N 2.35; found: C 52.11, H 3.67, N 2.30. IR (solid state): \u03bd\u02dc/cm\u22121=3116w, 3060w, 2979w, 2937w, 2160w\u2010br, 2007\u2005s (\u03bdCO), 1937\u2005s\u2010sh (\u03bdCO), 1836\u2005s (\u03bd\u03bc\u2010CO), 1596w, 1532\u2005s (\u03bdCN), 1506\u2005m\u2010sh, 1458w, 1434w, 1421w, 1381w, 1360w, 1341w, 1275w, 1247w, 1211w, 1179w, 1134w, 1075\u2005s\u2010sh, 1049\u2005s\u2010br, 1033\u2005s (\u03bdBF4), 1014\u2005s\u2010sh, 963w, 935w, 865w, 850w, 825w, 808\u2005m, 745\u2005s, 707\u2005m, 657\u2005m. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2021\u2005s (\u03bdCO), 1989w (\u03bdCO), 1836\u2005m (\u03bd\u03bc\u2010CO), 1538w (\u03bdCN). IR (MeCN): \u03bd\u02dc/cm\u22121=2023\u2005s (\u03bdCO), 1990w (\u03bdCO), 1837\u2005m (\u03bd\u03bc\u2010CO), 1537w (\u03bdCN). 1H NMR ([D6]acetone): \u03b4/ppm=8.7\u20138.3 , 8.30 , 8.16\u20138.09 , 8.1\u20137.7 , 7.75\u20137.70 , 5.68 , 5.19 , 4.87 , 1.51 . No changes were observed in the 1H spectrum after 14\u2005h at room temperature. 13C{1H} ([D6]acetone): \u03b4/ppm=325.2 (CN), 255.2 (\u03bc\u2010CO); 210.1, 209.8 (CO); 146.1 (C3); 134.3, 133.9 (C6+C11); 131.4 (C5); 129.5, 129.0 (C7+C10); 128.8, 128.6 (C8+C9); 126.3, 125.1 (C4+C12); 91.7, 91.2 (Cp); 66.8 (C1), 14.4 (C2). 19F{1H} NMR ([D6]acetone): \u03b4/ppm=\u2212151.0 (10BF4), \u2212151.1 (11BF4). 1H NMR (CDCl3): \u03b4/ppm=8.5\u20137.8 , 7.70\u20137.60 , 7.60\u20137.48 , 5.44 , 5.20\u20135.05 , 4.95 , 4.65 , 1.44 . 13C{1H} NMR (CDCl3): \u03b4/ppm=323.2 (br), 254.9, 209.2 (br), 208.4, ca. 145 (br); 133 (br), 132.9 (br); 130.1 (m), 128.1, 127.9, 126\u2013125 (m\u2010br), 124.0, 120.5, 117.1, 90.6, 90.3, 66 (br), 14.0. 19F{1H} NMR (CDCl3): \u03b4/ppm=\u2212150.9, \u2212151.0. The cis orientation of the Cp ligands in [4G]BF4 was ascertained by 1H NOE experiment in CDCl3. Thus, irradiation of one Cp resonance (\u03b4H 5.44 or 4.65\u2005ppm) evidenced a NOE interaction with the other.In a 150\u2005mL Schlenk tube under N[5H]Br. In a 100\u2005mL Schlenk tube under N2, benzyl bromide was added dropwise to a dark red solution of 1H (1.58\u2005mmol) in anhydrous MeCN (50\u2005mL) under vigorous stirring at 60\u2009\u00b0C. The mixture was stirred at reflux for 1.5\u2005h and conversion was checked by IR (MeCN). Next, the dark red solution was moved on top of an alumina column . Impurities were eluted with neat CH2Cl2, THF and MeCN, then a red band containing the title product was eluted with MeCN/MeOH . Volatiles were removed under vacuum (40\u2009\u00b0C), affording a red solid. Yield: 892\u2005mg, 92\u2009%. Compound [5H]Br is soluble in CH2Cl2, CHCl3, THF, iPrOH, poorly soluble in toluene, EtOAc, insoluble in Et2O. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2020s (\u03bdCO), 1988w (\u03bdCO), 1837\u2005m (\u03bd\u03bc\u2010CO), 1550w (\u03bdCN), 1534w. 1H NMR (CDCl3): \u03b4/ppm=7.48 , 7.40 , 7.30\u20137.26 , 5.69 , 5.56 , 5.52 .3SO3[5H]CF. In a 50\u2005mL round bottom flask, a solution of Ag(CF3SO3) in MeCN (5\u2005mL) was added to a dark red solution of [5H]I in MeCN (40\u2005mL) at 0\u2009\u00b0C. The mixture was stirred at 0\u2009\u00b0C for 1.5\u2005h while maintaining the system under protection from the light. The resulting suspension was filtered over celite and the filtrate solution was dried under vacuum. The residue was treated with hexane (10\u2005mL) then Et2O (50\u2005mL) and settled aside for 30\u2005min. Therefore, the solution was discarded and the washing procedure was repeated (x 2). The resulting red solid was dried under vacuum (40\u2009\u00b0C) and stored under N2 (slightly hygroscopic). Yield: 1076\u2005mg, 93\u2009%. Compound [5H]CF3SO3 is soluble in MeCN, acetone, MeOH, CH2Cl2, poorly soluble in Et2O, toluene, water and insoluble in hexane. Anal. calcd. for C29H24F3Fe2NO6S: C 50.98, H, 3.54, N, 2.05; found: C 50.59, H, 3.58, N, 2.26. IR (solid state): \u03bd\u02dc/cm\u22121=3106w, 3070\u20133230w, 2166w, 2013\u2005s (\u03bdCO), 1985\u2005s\u2010sh (\u03bdCO), 1827\u2005s (\u03bd\u03bc\u2010CO), 1588w, 1549\u2005m (\u03bdCN), 1533\u2005m, 1497w, 1454w, 1433w, 1420w, 1352w, 1260s (\u03bdSO3), 1223\u2005s\u2010sh (\u03bdSO3), 1153\u2005s (\u03bdSO3), 1096w, 1079w, 1029\u2005s, 858\u2005m, 767\u2005s, 737\u2005m\u2010sh, 698\u2005s. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=2023\u2005s (\u03bdCO), 1991\u2005m (\u03bdCO), 1840\u2005m (\u03bd\u03bc\u2010CO), 1550w (\u03bdCN), 1534w. IR (MeCN): \u03bd\u02dc/cm\u22121=2023\u2005s (\u03bdCO), 1990w (\u03bdCO), 1839\u2005m (\u03bd\u03bc\u2010CO), 1552w (\u03bdCN), 1538w. 1H NMR (CDCl3): \u03b4/ppm=7.50\u20137.44 , 7.44\u20137.39 , 7.20 , 5.66 , 5.50 , 5.40 . No changes were observed in the 1H spectrum after 2\u2005days at room temperature. 13C{1H} NMR (CDCl3): \u03b4/ppm=324.4 (CN), 253.7 (\u03bc\u2010CO), 208.2 (CO), 132.1 (C2), 129.9 (C4), 129.4 (C5), 127.8 (C3), 121.1 , 90.6 (Cp), 68.9 (C1). 19F{1H} NMR (CDCl3): \u03b4/ppm=\u221278.1\u2005ppm.2, a red suspension of [2C]CF3SO3 , Me3NO\u2009\u22c5\u20092H2O and LiCl in acetone (8\u2005mL) was stirred at reflux for 5\u2005h. Conversion was checked by IR (acetone) then volatiles were removed under vacuum. The brown residue was suspended in CH2Cl2 and moved on top of an alumina column , under N2. Impurities were eluted with CH2Cl2 then a brown band containing the title product was eluted with THF. The eluate was dried under vacuum and the residue was suspended hexane. The suspension was filtered; the resulting brown solid was washed with hexane and dried under vacuum (40\u2009\u00b0C). Yield: 55\u2005mg, 70\u2009%. A related one\u2010pot reaction with [2C]CF3SO3/Me3NO\u2009\u22c5\u20092H2O (1.6\u2005equiv)/NaCl (3.6\u2005equiv) in deaerated MeOH (8\u2005mL) at 40\u2009\u00b0C overnight gave 6 in 43\u2009% yield. A two\u2010step procedure comprising the [2C]CF3SO3/Me3NO\u2009\u22c5\u20092H2O (1\u2005equiv) reaction in MeCN at room temperature followed by LiCl (3.0\u2005equiv) in refluxing acetone for 3\u2005h gave 6 in 54\u2009% yield. Compound 6 is soluble in CH2Cl2, CHCl3, scarcely soluble in MeOH, insoluble in Et2O and hydrocarbons. Crystals of 6 for X\u2010ray analysis were obtained from a CHCl3 solution layered with pentane and settled aside at \u221220\u2009\u00b0C. IR (CH2Cl2): \u03bd\u02dc/cm\u22121=1977\u2005s (\u03bdCO), 1798\u2005s (\u03bd\u03bc\u2010CO), 1537\u2005m (\u03bdCN). IR (MeCN): \u03bd\u02dc/cm\u22121=1971\u2005s (\u03bdCO), 1795\u2005m (\u03bd\u03bc\u2010CO), 1539w (\u03bdCN). IR (solid state): \u03bd\u02dc/cm\u22121=3104w, 3070w, 1939\u2005s (\u03bdCO), 1782\u2005s (\u03bd\u03bc\u2010CO), 1558w\u2010sh, 1536\u2005s (\u03bdCN), 1456w, 1447w, 1419w, 1394\u2005m, 1360w, 1346w, 1317w, 1251w, 1182w, 1153w, 1116w, 1062\u2005m, 1016w, 1001w, 993\u2005m, 857w\u2010sh, 838\u2005m, 814\u2005m, 800s, 757\u2005s. 1H/13C NMR: mixture of isomers in 3\u2009:\u20092 ratio (1H CDCl3). Signals attributable to the minor isomer are italicized. 1H NMR (CDCl3): \u03b4/ppm=6.09, 5.01 ; 4.72, 4.70 ; 4.64, 4.62 ; 4.53, 4.05 ; 2.59, 2.37 ; 2.28\u20132.06, 1.98\u20131.69, 1.42\u20131.24 . No change in the 1H NMR spectrum was observed after 14\u2005h at room temperature. 13C{1H} NMR (CDCl3): \u03b4/ppm=334.8, 334.5 (CN); 268.1, 267.5 (\u03bc\u2010CO); 212.5, 211.9 (CO); 86.8, 86.6 (Cp); 86.4, 86.3 (Cp\u2019); 76.6, 75.4 (C2\u2010H); 44.4, 43.6 (C1\u2010H), 32.5, 32.1 (C3\u2010H); 32.3, 30.6, 26.3, 26.2, 26.0, 25.6, 25.6, 25.5 (C3\u2019\u2010H+C4\u2010H+C5\u2010H).In a 25\u2005mL Schlenk tube under N2A]CF3SO3, [2C]Cl\u2009\u22c5\u20092.34H2O, [2C]CF3SO3, [2D]CF3SO3 and 6 are reported in Table\u2005K\u03b1 radiation. The structures were solved by direct methods and refined by full\u2010matrix least\u2010squares based on all data using F2.2A]CF3SO3 which was located in the Fourier map and refined isotropically using the 1.2\u2010fold isoU of the parent N(2) atom. The H2O molecules of [2C]Cl\u2009\u22c5\u20092.34H2O are disordered and it was not possible to locate the hydrogen atoms. All non\u2010hydrogen atoms were refined with anisotropic displacement parameters. The crystals of 6 display a low quality, allowing the full determination of the overall connectivity and geometry of the complex, while the bonding parameters cannot be discussed in detail.Crystal data and collection details for BF4, [2C]Cl and [3F]CF3SO3 an inverse procedure was followed, starting from a solution of the compound in octanol\u2010saturated water. The partition coefficient was calculated as Pow=(A0aq\u2212Afaq)/Afaqwhere A0aqand Afaqare the absorbance in the aqueous phase before and after partition with the organic phase, respectively. The wavelength of the maximum absorption of each compound (320\u2013390\u2005nm range) was used for UV/Vis quantitation. The procedure was repeated three times for each sample (from the same stock solution); results are given as mean \u00b1 standard deviation and cell culture mediumStability in water in MeOH (0.50\u2005mL) then diluting with H2O up to 10.0\u2005mL total volume (5\u2009% v/v MeOH). Next, 3.20\u2005mL of the solution (nFe ca. 6\u2009\u22c5\u200910\u22123\u2005mmol) and a 7x2\u2005mm stir bar were transferred into a 4\u2010mL screw top vial (1.72\u2005mL neat gas phase volume). The mixture was sealed with a screw cap with a PTFE/silicone septum and heated at 37\u2009\u00b0C for 24\u2005h under stirring. Afterwards, the headspace was sampled with a gas tight microsyringe (250\u2005mL) and analysed by GC\u2010TCD. Therefore, the vial was vented, sealed and heated for further 24\u2005h, and GC\u2010TCD analysis was repeated. The amount of released CO was calculated based on a calibration curve obtained from analyses of known air/CO mixtures (1\u201310\u2009% v/v), and the number of equivalents released over a 24\u2005h period refers to the initial amount of compound (eqCO=nCO/nFe). The residual amount of starting diiron complex after 48\u2005h was calculated by assuming the release of 3 equivalents of CO per mole and the total amount of CO released. The procedure was repeated three times for each sample (from the same stock solution); results are given as mean \u00b1 standard deviation (Table\u2005S4).4OAc (1.25\u00d710\u22122\u2005M)cFe2=1\u00d710\u22123\u2005M) were prepared by dissolving the selected compound in DMSO (0.25\u2005mL) or MeOH , then diluting with HPLC water up to 5.0\u2005mL total volume. Next, aliquots of the Fe (0.50\u2005mL) and lysozyme (2.0\u2005mL) stock solutions were mixed and the final solution was kept at 37\u2009\u00b0C for 24\u2005h. The resulting suspensions were centrifuged to separate the orange\u2010brown precipitate. A blank solution (lysozyme only) was prepared and treated by the same procedure. Next, samples were diluted 1\u2009:\u200920 (v/v) with a water/MeCN solution containing 1\u2009% formic acid and analysed by HPLC\u2010MS and flow injection MS. For HPLC analyses, elution was conducted with a MeCN/water linear gradient (90\u2009:\u200910 to 5\u2009:\u200995 v/v over 60\u2005min), containing 0.05\u2009% trifluoroacetic acid. The following compounds were identified by their MS pattern: [2A]+ (tR=18.0\u2005min), [2B]+ (tR=13.8\u2005min), [2C]+ (tR=18.8\u2005min), [2D]+ (tR=17.0\u2005min), [2E]+ (tR=18.3\u2005min), [2F]+ (tR=9.7\u2005min), [3F]+ (tR=13.1\u2005min), [4G]+ (tR=23.0\u2005min), [5H]+ (tR=24.3\u2005min), lysozyme\u2010CH3 (tR=21.0\u2005min), lysozyme (tR=21.6\u2005min). In all cases, the starting organometallic cation was identified as the major Fe\u2010containing compound in solution. Protein peaks pattern were obtained following peak reconstruction. Peak integrals for lysozyme and methyl lysozyme were calculated from the extracted ion chromatograms from the flow injection MS analysis was stored at 4\u2009\u00b0C as received. A stock solution was prepared by dissolving the powder protein (125\u2005\u03bcM) and NH2B]BF4, [2C]CF3SO3, [2D]CF3SO3 and [3F]CF3SO3 were dissolved in water while the other compounds were dissolved in the minimum DMSO amount prior to cell culture testing. A calculated amount of the stock drug DMSO solution was added to the cell culture media to reach a final maximum DMSO concentration of 0.5\u2009%, which had no effects on cell viability.Compounds , cisplatin and ImmunoPure Cell cultures. Human colon (HCT\u201015), pancreatic (PSN\u20101) and breast (MCF\u20107) carcinoma cell lines along with human melanoma cells (A375) were obtained from American Type Culture Collection . Embryonic kidney HEK293 cells were obtained from the European Collection of Cell Cultures . Human ovarian 2008 cancer cells were kindly provided by Prof. G. Marverti . MCF\u20107 ADR cells were kindly provided by Prof. N. Colabufo . Cell lines were maintained in the logarithmic phase at 37\u2009\u00b0C in a 5\u2009% CO2 atmosphere using the following culture media containing 10\u2009% fetal calf serum , antibiotics (50\u2005units/mL penicillin and 50\u2005\u03bcg/mL streptomycin) and 2\u2005mM l\u2010glutamine: i) RPMI\u20101640 medium for PSN\u20101, 2008, HCT\u201015, MCF\u20107 and MCF\u20107 ADR cells; ii) DMEM for A375 and HEK293 cells.Spheroid cultures. Spheroid cultures were obtained by seeding 2.5\u00d7103 cells/well in round bottom non\u2010tissue culture treated 96 well\u2010plate in phenol red free RPMI\u20101640 medium , containing 10\u2009% FCS and supplemented with 20\u2009% methyl cellulose stock solution.MTT assay. The growth inhibitory effect towards tumour cells was evaluated by means of MTT assay as previously described.3 cells/well, dependent upon the growth characteristics of the cell line, were seeded in 96\u2010well microplates in growth medium (100 \u03bcL). After 24\u2005h, the medium was removed and replaced with a fresh one containing the compound to be studied at the appropriate concentration. Triplicate cultures were established for each treatment. After 24 or 72\u2005h, each well was treated with 10 \u03bcL of a 5\u2005mg/mL MTT saline solution, and following 5\u2005h of incubation, 100 \u03bcL of a sodium dodecyl sulfate (SDS) solution in HCl 0.01\u2005M were added. After an overnight incubation, cell growth inhibition was detected by measuring the absorbance of each well at 570\u2005nm using a Bio\u2010Rad 680 microplate reader. Mean absorbance for each drug dose was expressed as a percentage of the control untreated well absorbance and plotted against drug concentration. IC50 values, the drug concentrations that reduce the mean absorbance at 570\u2005nm to 50\u2009% of those in the untreated control wells, were calculated by the four\u2010parameter logistic (4\u2010PL) model. Evaluation was based on means from at least four independent experiments.Acid phosphatase (APH) assay. An APH modified assay was used for determining cell viability in 3D spheroids, as previously described.50 values were calculated with a four\u2010parameter logistic (4\u2010PL) model. All the values are the means \u00b1 SD of not less than four independent experiments.Mitochondrial membrane potential (\u0394\u03a8). The \u0394\u03a8 was assayed using the Mito\u2010ID\u00ae Membrane Potential Kit according to the manufacturer's instructions as previously described.3 per well) were seeded in 96\u2010well plates; after 24\u2005h, cells were washed with PBS and loaded with Mito\u2010ID detection reagent for 30\u2005min at 37\u2009\u00b0C in the dark. Afterwards, cells were incubated with increasing concentrations of tested complexes. Fluorescence intensity was estimated using a VICTOR X3 plate reader at \u03bbex=490 and \u03bbem=590\u2005nm. Antimycin (3\u2005\u03bcM) was used as positive control.ROS production. The production of ROS was measured in PSN\u20101 cells (104 per well) grown for 24\u2005h in a 96\u2010well plate in RPMI medium without phenol red (Merck). Cells were then washed with PBS and loaded with 10\u2005\u03bcM 5\u2010(and\u20106)\u2010chloromethyl\u20102\u2032,7\u2032\u2010dichlorodihydrofluorescein diacetate acetyl ester for 25\u2005min, in the dark. Afterwards, cells were washed with PBS and incubated with increasing concentrations of tested compounds. Fluorescence increase was estimated by using \u03bbex=485\u2005nm and \u03bbem=527\u2005nm in a VICTOR X3 (PerkinElmer) plate reader. Antimycin , a potent inhibitor of Complex III in the electron transport chain, was used as positive control.Intracellular CO release. PSN\u20101\u2005cell (1\u00d7104) were seeded in 96\u2010well plates. After 24\u2005h, cells were treated with 20\u2005\u03bcM of tested complexes or CORM\u2010401 (Merck) for 15\u2005min at 37\u2009\u00b0C and followed by 30\u2005min incubation with a fluorescent probe BioTracker Carbon Monoxide Probe 1 Live Cell Dye\u00ae (Merck). The intracellular fluorescence, indicative of CO accumulation in cells, was quantified using a VICTOR X3 (PerkinElmer) plate reader.Cellular uptake. PSN\u20101 and HEK293 cells (3\u00d7106) were seeded in 75\u2005cm2 flasks in growth medium (20\u2005mL). After overnight incubation, the medium was replaced and the cells were treated with tested compounds for 24\u2005h. Cell monolayers were washed twice with cold PBS, harvested and counted. Samples were than subjected to three freezing/thawing cycles at \u221280\u2009\u00b0C, and then vigorously vortexed. The samples were treated with highly pure nitric acid and transferred into a microwave Teflon vessel. Subsequently, samples were submitted to standard procedures using a speed wave MWS\u20103 Berghof instrument . After cooling, each mineralized sample was analysed for iron using a Varian AA Duo graphite furnace atomic absorption spectrometer (Varian) at the wavelength of 248.3\u2005nm. The calibration curve was obtained using known concentrations of standard solutions purchased from Sigma Chemical Co.Statistical analysis. All values are the means \u00b1 SD of no less than three measurements starting from three different cell cultures. Multiple comparisons were made by ANOVA followed by the Tukey\u2010Kramer multiple comparison test , using GraphPad software.50 values after 24\u2005h of incubation; MS spectra from lysozyme interaction experiments. Deposition numbers 2A]CF3SO3), 2045778 (for [2C]CF3SO3), 2045777 (for [2C]Cl), 2045780 (for [2D]CF3SO3) and 2045781 (for 6)2045779 should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "The mol\u00adecular and crystal structure of a ferrocenyl derivative with an undecyl-1,11-diol chain on one cyclo\u00adpenta\u00addienyl ring is reported: O\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22ef\u03c0(ring) contacts occur in the extended structure. 5H5)(C16H27O2)], comprises an \u03b1,\u03c9-diol-substituted undecyl chain with a ferrocenyl substituent at at one terminus. The alkane chain is inclined to the substituted ring of the ferrocene grouping by 84.22\u2005(13)\u00b0. The ferrocene rings are almost eclipsed and parallel. The crystal structure features O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 contacts that stack the mol\u00adecules along the c-axis direction. A Hirshfeld surface analysis reveals that H\u22efH inter\u00adactions (83.2%) dominate the surface contacts.The racemic title compound, [Fe(C It was synthesized to provide a ferrocenyl-substituted diol for the preparation of polyesters with regular pendant electroactive groups. Similar ferrocenyl neo-pentyl diol-derived terephthalate polymers have been shown to display inter\u00adesting electrochemical properties using LiAlH4. Enanti\u00adomeric selection of the individual chiral forms should be possible using more complex synthetic methodology (C16H27O2)], comprises a ferrocene unit that carries a well-ordered undecane chain (atoms C11\u2013C21) with hydroxyl substituents at the 1 and 11 positions along the chain \u00b0 between them. The C11 undecyl chain in 1 is conformationally extended with the typical anti\u00adperiplanar \u2005\u00c5. The C1\u2013C5 and C6\u2013C10 cyclo\u00adpenta\u00addienyl rings of the ferrocenyl group are approximately 3\u00b0 from being eclipsed and are almost coplanar with a dihedral angle of 1.7\u2005(2)\u00b0 between them; the separation of the ring centroids is is 3.298\u2005(2)\u2005\u00c5.The title compound, , 1.7\u20131.3 . 13C NMR (CDCl3): 94.7 (Fc ipso), 69.7 (\u2013CHOH\u2013), 68.3 (Cp), 67.9, 67.7, 67.3, 65.2 (Fc\u2014C\u03b1 & \u03b2), 63.2 (\u2013CH2OH), 38.3, 32.9, 29.6, 29.6, 29.5, 29.5, 26.1, 25.8 (\u2013CH2\u2013). UV\u2013vis (CH2Cl2): 325 (90), 440 (110) nm (\u025b).The title compound iso(H) = 1.5 Ueq(O). All H-atoms bound to C were refined using a riding model with C\u2014H = 0.95\u20131.00\u2005\u00c5 and Uiso(H) = 1.2Ueq(C). Despite repeated attempts to grow crystals of better quality, the crystals obtained were weakly diffracting and the extent of diffraction observed is poor with sin\u2005(\u03b8max)/\u03bb = 0.544 (2\u03b8max = 44.5\u00b0). Despite this, the structure solved and refined adequately.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698902101358X/hb8007sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902101358X/hb8007Isup2.hklStructure factors: contains datablock(s) I. DOI: 2130725CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, dimeric C\u2014H\u22efO hydrogen bonds connect pairs of cations, producing two 30H28NO2+\u00b7CF3O3S\u2212, the four tetra\u00adhydro\u00adfuran rings adopt envelope conformations. In the crystal, pairs of cations are linked by dimeric C\u2014H\u22efO hydrogen bonds, forming two R22(6) ring motifs parallel to the (001) plane. The cations and anions are connected by further C\u2014H\u22efO hydrogen bonds, forming a three-dimensional network structure. Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from H\u22efH (47.6%), C\u22efH/H\u22efC (20.6%), O\u22efH/H\u22efO (18.0%) and F\u22efH/H\u22efF (9.9%) inter\u00adactions.In the cation of the title salt, C The mol\u00adecular conformation of the cation is stabilized by weak intra\u00admolecular C21\u2014H21B\u22efO12 and C21\u2014H21B\u22efO13 contacts (Table\u00a01B/C11C/C3A/C3) in the cation exhibits a chair conformation . The benzene ring (C7A/C8\u2013C11/C11A) fused with the central tetra\u00adhydro\u00adfuran ring makes dihedral angles of 53.43\u2005(5) and 58.64\u2005(5)\u00b0, respectively, with the C22\u2013C27 and C32\u2013C37 phenyl rings of the benzyl groups attached to the N atom. These phenyl rings make a dihedral angle of 73.81\u2005(5)\u00b0 with each other.In the cation \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01], forming two et al., 1995A\u22efO1i, C6\u2014H6A\u22efO12ii, C6A\u2014H6AA\u22efO3ii, C7\u2014H7A\u22efO13ii, C31\u2014H31A\u22efO2i and C9\u2014H9A\u22efCg10iii hydrogen bonds, forming a three-dimensional network inter\u00adactions constitute the primary factor in the crystal packing, with C\u22efH/H\u22efC , O\u22efH/H\u22efO and F\u22efH/H\u22efF inter\u00adactions constituting the next stronger contributions. Numerical values of these inter\u00adactions together with other percentage contributions of weaker inter\u00adactions are compiled in Table\u00a03The inter\u00admolecular inter\u00adactions Table\u00a02 were qu\u00ad\u22efC Fig.\u00a08c, O\u22efH/H\u22efO Fig.\u00a08d and F\u22ef\u22efF Fig.\u00a08e inter\u00adet al., 2016viz. IQOTOA and ac plane. Additionally, C\u2014H\u22efO hydrogen bonds form a C(6) chain, linking the mol\u00adecules along the b-axis direction. In the crystal of ERIVIL, mol\u00adecules are connected into b axis. In MIGTIG, the mol\u00adecules are linked only by weak van der Waals inter\u00adactions.IQOTOA, OMUTAU and OMEMAX each crystallize with two mol\u00adecules in the asymmetric unit. In the crystal, mol\u00adecule pairs generate centrosymmetric rings with -2,2-Dibenzyl-2,3,6a,11c-tetra\u00adhydro-1H,6H,7H-3a,6:7,11b-di\u00adepoxy\u00addibenzoisoquinolin-2-ium tri\u00adfluoro\u00admethane\u00adsulfonate (1)3CN (20\u2005ml). Then an equivalent of 2-(tri\u00admethyl\u00adsil\u00adyl)phenyl tri\u00adfluoro\u00admethane\u00adsulfonate was added to the solution under an argon atmosphere. The mixture was refluxed for 4\u2005h . After one more portion of 2-(tri\u00admethyl\u00adsil\u00adyl)phenyl tri\u00adfluoro\u00admethane\u00adsulfonate and CsF had been added to the mixture, all procedures were repeated. After the mixture was cooled to room temperature, the CsF was filtered off through a thin layer of SiO2, and the resulting solution was concentrated under reduced pressure. The residue (yellow oil) turned out to be a multicomponent mixture. It was separated using column chromatography on silica gel. The least mobile fraction represented the target product, 1. In addition, two by-products 2 (12%) and 3 (17%) were isolated. Single crystals of 1 were obtained by slow crystallization from ethyl acetate.Cesium fluoride (CsF) was added to benzyl\u00adbis\u00ad(furan-2-ylmeth\u00adyl)amine (0.0022\u2005mol) dissolved in dry CHUiso(H) = 1.2Ueq(C). Five reflections I. DOI: 10.1107/S2056989021010173/wm5617Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the title mol\u00adecules are linked by inter\u00admolecular C\u2014H\u22efN, C\u2014H\u22efCl, C\u2014H\u22ef\u03c0 contacts and \u03c0\u2013\u03c0 stacking inter\u00adactions. A Hirshfeld surface analysis was undertaken to qu\u00adantify the inter\u00admolecular inter\u00adactions. 24H20ClNO2, the mean planes of 4-chloro\u00adphenyl, 2-methyl\u00adphenyl and phenyl\u00adene rings make dihedral angles of 62.8\u2005(2), 65.1\u2005(3) and 15.1\u2005(2)\u00b0, respectively, with the 5-methyl-1,2-oxazole ring. In the crystal, mol\u00adecules are linked by inter\u00admolecular C\u2014H\u22efN, C\u2014H\u22efCl, C\u2014H\u22ef\u03c0 contacts and \u03c0\u2013\u03c0 stacking inter\u00adactions between the phenyl\u00adene groups. Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from H\u22efH (48.7%), H\u22efC/C\u22efH (22.2%), Cl\u22efH/H\u22efCl (8.8%), H\u22efO/O\u22efH (8.2%) and H\u22efN/N\u22efH (5.1%) inter\u00adactions.In the title compound, C SCXD has thus become an essential tool for drug development to unambiguously determine the three-dimensional structures of mol\u00adecules, which eventually paves the way for rapid development of new mol\u00adecules = 3.958\u2005(2)\u2005\u00c5] to 1.5787\u2005\u00c5 (blue) Fig.\u00a04.dnorm values on the surface, correspond to the C\u2014H\u22efN (C17\u2014H17A\u22efN1), C\u2014H\u22efCl (C8\u2014Cl1\u22efH1C\u2014C1) and C\u2014H\u22ef\u03c0 (C6\u2014H6\u22efphenyl\u00adene) inter\u00adactions shows the total two-dimensional fingerprint plot providing information on the major and minor percentage contributions of the inter\u00adatomic contacts to the Hirshfeld surface of the title compound. The blue colour refers to the frequency of occurrence of the pair and the grey colour is the outline of the full fingerprint show that the H\u22efH contacts clearly make the most significant contribution to the Hirshfeld surface (48.7%). The H\u22efC/C\u22efH, Cl\u22efH/H\u22efCl, H\u22efO/O\u22efH and H\u22efN/N\u22efH contacts contribute 22.2, 8.8, 8.2 and 5.1%, respectively . The remaining weaker contacts are listed in Table\u00a03Fig.\u00a06ly Fig.\u00a06c\u2013f. Theet al., 2015The large number of H\u22efH, H\u22efC/C\u22efH, Cl\u22efH/H\u22efCl, H\u22efO/O\u22efH and H\u22efN/N\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions play the major roles in the crystal packing (Hathwar E)-2- (di\u00admethyl\u00adamino)\u00adethen\u00adyl]-1,2-oxazole-4- carboxyl\u00adate , whereas in mol\u00adecule B, it points back towards the benzene ring [C\u2014C\u2014C=O = 17.9\u2005(4)\u00b0]. The dihedral angles between the oxazole and benzene rings are also somewhat different . Each mol\u00adecule features an intra\u00admolecular C\u2014H\u22efO inter\u00adaction, which closes an S(6) ring. In the crystal, the B mol\u00adecules are linked into C(12) chains along the c-axis direction by weak C\u2014H\u22efCl inter\u00adactions. In the crystal of (II), the components are linked by O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds, where the water mol\u00adecule acts as both an H-atom donor and an acceptor, into a tape along the a-axis direction with an In compound (I)N-hy\u00addroxy-4-[(2-methyl\u00adbenz\u00adyl)\u00adoxy]benzimidoyl chloride in diethyl ether (6\u2005ml) was added Et3N . The resulting mixture was stirred for 2\u2005h in an ice bath, and the precipitate formed was filtered off. The filtrate was evaporated under vacuum to obtain the aryl\u00adnitriloxide inter\u00admediate.Step 1: To a solution of 2SO4, filtered and evaporated to dryness. The crude product was purified by automated-flash chromatography on silica gel (12\u2005g) eluting with a gradient of 0 to 40% EtOAc in hexane. The obtained pure product was recrystallized from methanol. Crystals for structural study were obtained by slow cooling of the solution, yield 77%, m.p. 387.2\u2013388.6\u2005K.Step 2: To a solution of NaH in dry THF (4\u2005ml), 4-chloro\u00adphenyl\u00adacetone was added dropwise, and stirred for 1\u2005h under a nitro\u00adgen atmosphere in an ice bath. At the end of the period, the aryl\u00adnitriloxide inter\u00admediate was dissolved in dry THF (4\u2005ml), and was added to the reaction mixture, then stirred at room temperature overnight. Upon completion of the reaction, aqueous ammonium chloride solution was added, and the product was extracted with EtOAc (2 \u00d7 50\u2005mL). The combined organic extracts were dried over anhydrous Na1H NMR : \u03b4 2.29 , 2.39 , 5.07 , 7.05 , 7.15\u20137.25 , 7.27 , 7.38 , 7.47 .13C NMR : \u03b4 11.21, 18.42, 67.98, 113.96, 114.97, 120.84, 125.77, 128.15, 128.59, 128.86, 129.43, 130.12, 131.44, 132.57, 134.58, 136.64, 159.49, 160.09, 166.93. HRMS (m/z): [M + H]+ calculated for C24H21ClNO2: 390.1261; found: 390.1263.Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl). In the final refinement, three outliers I. DOI: 10.1107/S2056989021002383/yk2147Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021002383/yk2147Isup3.cmlSupporting information file. DOI: 2067449CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "This study was designed to identify the fluid spaces that are most changed during ultrafiltration (UF) according to intradialytic blood pressure (BP) difference. BP data were collected five times . Intradialytic BP difference was calculated as the highest minus lowest of these BP measurements. Intradialytic systolic BP (SBP) difference over 20\u2009mm Hg and diastolic BP (DBP) difference over 10\u2009mm Hg were defined as wide intradialytic SBP difference (SYS-W) and DBP difference (DIA-W), respectively. We measured the various fluid spaces before HD and 1\u20134\u2009h of HD, and 30\u2009min after HD using a portable, whole-body bioimpedance spectroscopy (BIS). In this study, 85 prevalent patients aged over 18\u2009years with a fixed dry weight , undergoing HD had participated. 1) Mean relative reduction of extracellular water (ECW) was significantly higher in SYS-W than in narrow intradialytic SBP difference (SYS-N) patients from 1\u2009h to 30\u2009min after HD. 2) Mean relative reduction of intracellular water (ICW) was significantly lower in DIA-W than in narrow intradialytic DBP difference (DIA-N) patients from 1\u2009h to 30\u2009min after HD. 3) ECW of patients with SYS-W was significantly lower than that of patients with SYS-N. Patients with SYS-W have the characteristics of fluid shifts in which reduction of ECW was steeper than patients with SYS-N whereas fluid shifts of ICW were lower in patients with DIA-W than patients with DIA-N. A recent large, retrospective cohort study observed first-year mortality to be higher in HD patients with BP variability than in those with stable BP, independent of the absolute BP level, suggesting that BP variability is associated with worse outcomes at all levels of BP . The majvs. intracellular [IC]) source of fluid shifts during the UF procedure and the relative ratio of these compartments during conventional renal replacement therapy in patients with asymptomatic intradialytic BP variability. An authoritative journal accepted the definition of BP variability by the difference between the highest and lowest BP during each session , patients with a history of ischemic or hemorrhagic stroke, or heart disease including atrial fibrillation < 45% by 2-D echocardiography); those with peripheral arterial occlusive disease, such as gangrene, and amputees; and those with infection or hemiplegia were excluded. Some of the patient\u2019s characteristics are given in 2 hollow fiber poly-sulfone membranes, blood flow rates of 250\u2013280\u2009mL/min, and a dialysate flow rate of 500\u2009mL/min, using the Fresenius 5008S machine . The mean UF volume was 2457.65\u2009\u00b1\u2009947.73\u2009mL, and treatment time was 4\u2009h. The dialysis fluid contained 138 mEq/L sodium, 2 mEq/L potassium, 2.5 mEq/L calcium, 1.0 mEq/L magnesium, 108.5 mEq/L chloride, 35 mEq/L bicarbonate, and 99.1\u2009mg/dL glucose, and the temperature was maintained at 36\u2009\u00b0C.All patients underwent high-flux HD thrice weekly with a bicarbonate buffer for 4\u2009h, using 1.0\u20131.4-mBP data were collected five times (before HD and 1\u20134\u2009h of HD), and the highest and lowest measurements were identified. Intradialytic BP difference was calculated as the highest minus lowest BP measurements of each dialysis session . Intradiurea) in liters (L). The BCM determines resistance and reactance at 50 discrete frequencies from 3 to 1000\u2009kHz. ECW and ICW resistance are obtained on the basis of a Cole model [BCM measurement has been recommended that subjects should be resting in a supine position for at least 5\u2009min, before the BCM-measurement is started, such that fluid volume equilibration has taken place . We measle model . Using tle model .Because of the small number of patients, normal distribution was tested using a single sample Kolmogorov\u2013Smirnov analysis. Continuous variables are expressed as mean\u2009\u00b1\u2009SD or median (range) according to normal distribution of data. Categorical variables are expressed as percentage (%). Between-group differences were assessed for significance using a Student t-test for normal distribution, Mann\u2013Whitney U tests for skewed distribution, or Chi-Square test for categorical variables. Statistical analyses were performed using SPSS software version 14.0 .The study included a total of 85 patients undergoing HD (vintage of 70.98\u2009\u00b1\u200959.47\u2009months), 46 (54.12%) men with a mean age of 65.38\u2009\u00b1\u200912.45\u2009years. Here, 44 (51.76%) patients had diabetes mellitus and 50 (58.82%) patients had hypertension. The anti-hypertensive medications administered by the patients included angiotensin-converting enzyme inhibitor by nine patients (10.59%), angiotensin II receptor blocker by 18 patients (21.18%), calcium channel blocker by 30 patients (35.29%), alpha blocker by three patients (3.53%), beta-blocker by 21 patients (24.71%), diuretic by seven patients (8.24%), and other anti-hypertensive medications by 2 patient (2.35%). In 62 (72.94%) or 35 (41.17%) of the patients on HD, phosphorus binders or active vitamin D metabolites were administered, respectively. Other variables are summarized in vs. 0.984\u2009\u00b1\u20090.027; 2\u2009h, 0.988\u2009\u00b1\u20090.047 vs. 0.975\u2009\u00b1\u20090.034; 3\u2009h, 0.970\u2009\u00b1\u20090.032 vs. 0.958\u2009\u00b1\u20090.032; 4\u2009h, 0.960\u2009\u00b1\u20090.035 vs. 0.948\u2009\u00b1\u20090.035, 30\u2009min after HD; 0.955\u2009\u00b1\u20090.029 vs. 0.941\u2009\u00b1\u20090.032, respectively) .vs. 0.977\u2009\u00b1\u20090.019*; 2\u2009h, 0.966\u2009\u00b1\u20090.027 vs. 0.952\u2009\u00b1\u20090.026*; 3\u2009h, 0.942\u2009\u00b1\u20090.032 vs. 0.924\u2009\u00b1\u20090.033*; 4\u2009h, 0.923\u2009\u00b1\u20090.037 vs. 0.901\u2009\u00b1\u20090.038*, 30\u2009min after HD; 0.916\u2009\u00b1\u20090.038 vs. 0.895\u2009\u00b1\u20090.042*, respectively, *p\u2009<\u2009.05) .vs. 0.991\u2009\u00b1\u20090.052; 2\u2009h, 1.008\u2009\u00b1\u20090.081 vs. 0.996\u2009\u00b1\u20090.064; 3\u2009h, 0.996\u2009\u00b1\u20090.047 vs. 0.989\u2009\u00b1\u20090.054; 4\u2009h, 0.997\u2009\u00b1\u20090.044 vs. 0.991\u2009\u00b1\u20090.058, 30\u2009min after HD; 0.992\u2009\u00b1\u20090.038 vs. 0.987\u2009\u00b1\u20090.049, respectively) .vs. 0.986\u2009\u00b1\u20090.024; 2\u2009h, 0.971\u2009\u00b1\u20090.025 vs. 0.982\u2009\u00b1\u20090.040; 3\u2009h, 0.955\u2009\u00b1\u20090.027 vs. 0.964\u2009\u00b1\u20090.036; 4\u2009h, 0.945\u2009\u00b1\u20090.034 vs. 0.953\u2009\u00b1\u20090.036, 30\u2009min after HD; 0.938\u2009\u00b1\u20090.026 vs. 0.947\u2009\u00b1\u20090.036, respectively) .vs. 0.975\u2009\u00b1\u20090.021; 2\u2009h, 0.959\u2009\u00b1\u20090.021 vs. 0.953\u2009\u00b1\u20090.030; 3\u2009h, 0.932\u2009\u00b1\u20090.027 vs. 0.926\u2009\u00b1\u20090.038; 4\u2009h, 0.910\u2009\u00b1\u20090.035 vs. 0.905\u2009\u00b1\u20090.042, 30\u2009min after HD; 0.901\u2009\u00b1\u20090.036 vs. 0.898\u2009\u00b1\u20090.046, respectively) .vs. 0.997\u2009\u00b1\u20090.049*; 2\u2009h, 0.982\u2009\u00b1\u20090.043 vs. 1.009\u2009\u00b1\u20090.073*; 3\u2009h, 0.977\u2009\u00b1\u20090.045 vs. 1.001\u2009\u00b1\u20090.053*; 4\u2009h, 0.980\u2009\u00b1\u20090.056 vs. 0.999\u2009\u00b1\u20090.050*, 30\u2009min after HD; 0.975\u2009\u00b1\u20090.040 vs. 0.995\u2009\u00b1\u20090.050, respectively, *p\u2009<\u2009.05) .vs. 0.993\u2009\u00b1\u20090.021; 2\u2009h, 0.977\u2009\u00b1\u20090.021 vs. 0.977\u2009\u00b1\u20090.016; 3\u2009h, 0.965\u2009\u00b1\u20090.024 vs. 0.966\u2009\u00b1\u20090.029; 4\u2009h, 0.953\u2009\u00b1\u20090.020 vs. 0.953\u2009\u00b1\u20090.025, 30\u2009min after HD; 0.950\u2009\u00b1\u20090.021 vs. 0.950\u2009\u00b1\u20090.030, respectively) .vs. 0.977\u2009\u00b1\u20090.019*; 2\u2009h, 0.969\u2009\u00b1\u20090.014 vs. 0.949\u2009\u00b1\u20090.026*; 3\u2009h, 0.946\u2009\u00b1\u20090.018 vs. 0.923\u2009\u00b1\u20090.034*; 4\u2009h, 0.924\u2009\u00b1\u20090.018 vs. 0.904\u2009\u00b1\u20090.038*, 30\u2009min after HD; 0.922\u2009\u00b1\u20090.019 vs. 0.885\u2009\u00b1\u20090.043*, respectively, *p\u2009<\u2009.05) .vs. 1.014\u2009\u00b1\u20090.050*; 2\u2009h, 0.983\u2009\u00b1\u20090.039 vs. 1.010\u2009\u00b1\u20090.034*; 3\u2009h, 0.982\u2009\u00b1\u20090.042 vs. 1.015\u2009\u00b1\u20090.037*; 4\u2009h, 0.982\u2009\u00b1\u20090.034 vs. 1.012\u2009\u00b1\u20090.038*, 30\u2009min after HD; 0.984\u2009\u00b1\u20090.036 vs. 1.010\u2009\u00b1\u20090.039*, respectively, *p\u2009<\u2009.05) .urea, TBW, ICW, and E/I ratio were no significant difference between the groups weight, UFV, Ve groups .The main findings of our study are 1) changes in the ECW of patients with wide intradialytic SBP difference were significantly steeper than those of patients with narrow intradialytic SBP difference from 1\u2009h to 30\u2009min after HD. 2) there were no significant fluid shifts in the ICW of patients with wide intradialytic DBP difference, whereas that of patients with narrow intradialytic DBP difference was significantly decreased from 1\u2009h to 30\u2009min after HD. 3) ECW of patients with wide intradialytic SBP difference was significantly lower than that of patients with narrow intradialytic SBP difference.The results of the study suggest that maintenance of intradialytic SBP stability is dependent on the amount of ECW reduction during HD and that DBP stability is associated with the shifts of ICW. That is because there was no difference in the changes of TBW, whereas the difference in ECW or ICW changes could be observed between patients with narrow SBP difference and wide SBP difference, between patients with narrow DBP difference and wide DBP difference during HD, respectively. This phenomenon has been confirmed from patients with narrow intradialytic SBP/DBP difference, once again. In particular, ICW in patients with narrow intradialytic SBP/DBP difference showed a significant decrease, whereas that in patients with wide intradialytic SBP/DBP difference demonstrated no significant reduction of ICW from 1\u2009h of HD compared to the start of HD. The above changes were continued until 30\u2009min after HD without further reduction or increase. This indicates that the amount of shifts of ICW to ECW may affect the maintenance of SBP and DBP stability in situations where ECW is continuously reduced after HD. Thus, in contrast to a previous report , fluid rA recently introduced equipment called a BCM utilizes a well-established technical solution to assess the absolute volume of the body fluid distribution using 50 multiple, discrete frequencies from 3 to 1000\u2009kHz . DespiteThe clinical dilemmas and prognostic uncertainties exist in a patient with asymptomatic intradialytic BP falls, elevations, and fluctuations compared to overt intradialytic BP abnormalities such as IDH or IH. This absence of associated symptoms contributes to the tendency to regard asymptomatic BP fluctuations as \u2018normal\u2019 BP. However, there is a possibility that aberrant, asymptomatic intradialytic BP changes induce harmful effects to HD patients. Thus, our study was designed to evaluate factors that may influence the fluid shifts in asymptomatic patents with intradialytic BP falls. But, there were no significant differences between the hydration status, presence of DM, age, sex, HT medication, HD vintage, and fluid distribution index. Only we found that ECW of patients with wide intradialytic SBP difference was significantly lower than that of patients with narrow intradialytic SBP difference. Unfortunately, it would be difficult to explain the exact meaning of our results. Nevertheless, further study should be needed to elucidate the changes of IVF and ISF absolute values and relevant factors in asymptomatic patients with wide intradialytic BP difference.per se and a relation between a larger decline in ECW and BP is to be expected. If we repeated the analysis with additional BP variability metrics such as the BP residual in a random-effects model [This study had some limitations. First, we used intradialytic BP difference as an index of BP fluctuation instead of other intradialytic BP variability index, such as standard deviation, absolute value of the difference between successive BP measurements and BP residual. However, this reflects more the BP drop during HD than the variability ts model , we thouPatients with wide intradialytic SBP difference have the characteristics of fluid shifts in which reduction of ECW was steeper than patients with narrow intradialytic SBP difference whereas fluid shifts of ICW were lower in patients with wide intradialytic DBP difference than patients with narrow intradialytic DBP difference. This phenomenon has been confirmed from patients with narrow intradialytic SBP/DBP difference."} +{"text": "Scientific Reports 10.1038/s41598-021-85129-1, published online 10 March 2021Correction to: The original version of this Article contained errors.Author Fatih Semerci was incorrectly given as Faith Semerci.In addition, in the Introduction,\u201cThe NSC-6 Ab, which produced the most robust staining of NPCs, corresponded to Brain-Associated Signal Protein 1 (BASP1), a protein not previously described in NPCs.\u201dnow reads:\u201cThe NSC-6 Ab, which produced the most robust staining of NPCs, corresponded to Brain-Abundant, membrane-attached Signal Protein 1 (BASP1), a protein not previously described in NPCs.\u201dIn the Results section, under subheading \u201cNSC-6 stains mouse and human NPCs\u201d,\u201cIn addition, we derived human neuroprogenitor cells (hNPCs) from inducible pluripotent stem cells (iPSCs) obtained from a healthy adult and stained them with NSC-6.\u201dnow reads:\u201cIn addition, we derived human neuroprogenitor cells (hNPCs) from induced pluripotent stem cells (iPSCs) obtained from a healthy adult and stained them with NSC-6.\u201dUnder the subheading \u201cNSC-6-labeled BASP1 is regulated temporally in the mammalian brain\u201d,\u201cConsistent with the data obtained in mice, our results show high BASP1 expression in the hippocampus, the brainstem, and the spinal cord .\u201dnow reads:\u201cConsistent with the data obtained in mice, our results show high BASP1 expression in the human hippocampus, the brainstem, and the spinal cord .\u201dIn the Materials and methods section, under subheading \u201cEmbryonic neurosphere culture and hybridoma production\u201d,5\u00a0cells/10\u00a0ml of proliferation media containing Neurocult Basal Media (STEM CELL TECHNOLOGIES), 10% Proliferation Supplement (STEM CELL TECHNOLOGIES), 20\u00a0ng/ml EGF and FGF (SIGMA) and 1% antibiotic\u2013antimycotic (GIBCO).\u201d\u201cWhole brains from C57/BL6 embryonic day 12 (E12) mice were dissected, digested with collagenase for 2\u00a0h at 37\u00a0\u00b0C, filtered twice through 0.4\u00a0\u03bcm filters and plated at 3\u2009\u00d7\u200910now reads:5\u00a0cells/10\u00a0ml of proliferation media containing Neurocult Basal Media (STEM CELL TECHNOLOGIES), 10% Proliferation Supplement (STEM CELL TECHNOLOGIES), 20\u00a0ng/ml EGF and FGF (SIGMA) and 1% antibiotic\u2013antimycotic (GIBCO).\u201d\u201cWhole brains from C57/BL6 embryonic day 12 (E12) mice were dissected, digested with collagenase for 2\u00a0h at 37\u00a0\u00b0C, filtered twice through 40\u00a0\u03bcm filters and plated at 3\u2009\u00d7\u200910Under subheading \u201cTwo-dimensional electrophoresis\u201d,50.\u201d\u201cBriefly, IPG strips were passively equilibrated under mineral oil for 18\u00a0h at 23\u00a0\u00b0C with 90\u00a0\u00b5g of solubilized protein in 200\u00a0\u00b5l STnow reads:50.\u201d\u201cBriefly, IPG strips were passively equilibrated under mineral oil for 18\u00a0h at 23\u00a0\u00b0C with 90\u00a0\u00b5g of solubilized protein in 200\u00a0\u00b5l STUnder the subheading \u201cNeurosphere culture from adult brain SVZ\u201d,\u201c3\u00a0ml of Trypsin inhibitor (from glycine max) was added and gently mixed and the tissue suspension was passed through 0.7\u00a0\u03bcm pore size filter followed by centrifuge at 700\u00a0rpm for 5\u00a0min. The supernatant was removed and the pellet was resuspended in the NSC culture media .\u201dnow reads:\u201c3\u00a0ml of Trypsin inhibitor (from glycine max) was added and gently mixed and the tissue suspension was passed through 70\u00a0\u03bcm pore size filter followed by centrifuge at 700\u00a0rpm for 5\u00a0min. The supernatant was removed and the pellet was resuspended in the NSC culture media .\u201dUnder the subheading, \u201cHuman brain organoids and hNPCs\u201d,\u201cSpecifically, iPSCs colonies were first dissociated into single cells (D0) with Accutase (SIGMA-ALDRICH) and 1.5 million cells seeded per well of an AggreWell00 plate (STEM CELL TECHNOLOGIES) in a medium containing KnockOut DMEM, 20% KnockOut Serum Replacement, 1% penicillin and streptomycin (P/S) solution, 0.5X GlutaMAX, 1\u00d7 MEM Non-Essential Amino Acids , 5\u00a0\u00b5M Dorsomorphin (PEPROTECH), 10\u00a0\u00b5M SB431542 (PEPROTECH), 100\u00a0\u00b5M 2-Mercaptoethanol (SIGMA), and 10\u00a0\u00b5M Y-27632 (TOCRIS).\u201dnow reads:\u201cSpecifically, iPSCs colonies were first dissociated into single cells (D0) with Accutase (SIGMA-ALDRICH) and 1.5 million cells seeded per well of an AggreWell800 plate (STEM CELL TECHNOLOGIES) in a medium containing KnockOut DMEM, 20% KnockOut Serum Replacement, 1% penicillin and streptomycin (P/S) solution, 0.5X GlutaMAX, 1\u00d7 MEM Non-Essential Amino Acids , 5\u00a0\u00b5M Dorsomorphin (PEPROTECH), 10\u00a0\u00b5M SB431542 (PEPROTECH), 100\u00a0\u00b5M 2-Mercaptoethanol (SIGMA), and 10\u00a0\u00b5M Y-27632 (TOCRIS).\u201dLastly, Reference 11 was incorrectly given as:Front. Biol. (Beijing)\u00a011, 151\u2013167 (2016).Semerci, F. & Maleic-Savatic, M. Transgenic mouse models for studying adult neurogenesis.\u00a0The correct reference is listed below:Front. Biol. (Beijing)\u00a011, 151\u2013167 (2016).Semerci, F. & Maletic-Savatic, M. Transgenic mouse models for studying adult neurogenesis.\u00a0The original Article has been corrected."} +{"text": "The title mol\u00adecule adopts a conformation with the two phenyl substituents disposed on opposite sides of the mean plane of the iso\u00adquinoline unit. In the crystal, corrugated layers of mol\u00adecules are formed by N\u2014H\u22efO, C\u2014H\u22efN and C\u2014H\u22efS hydrogen bonds together with C\u2014H\u22ef\u03c0(ring) inter\u00adactions. These layers are connected by C\u2014H\u22efO contacts. 29H29N3O4S, adopts a conformation with the two phenyl substituents disposed on opposite sides of the mean plane of the iso\u00adquinoline unit. In the crystal, corrugated layers of mol\u00adecules are formed by N\u2014H\u22efO, C\u2014H\u22efN and C\u2014H\u22efS hydrogen bonds together with C\u2014H\u22ef\u03c0(ring) inter\u00adactions. These layers are connected by C\u2014H\u22efO contacts. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (45.2%), C\u22efH/H\u22efC (20.2%), O\u22efH/H\u22efO (15.8%) and N\u22efH/H\u22efN (11.0%) inter\u00adactions.The title mol\u00adecule, C In AKIVUO, a layer structure with the layers parallel to . In POPYEB, mol\u00adecules are packed in a herringbone manner parallel to (103) and (10via weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0(ring) inter\u00adactions. In ENOCIU, various C\u2014H\u22ef\u03c0 and C\u2014H\u22efO bonds link the mol\u00adecules together. In NIWPAL, the mol\u00adecules are linked by N\u2014H\u22efO inter\u00admolecular hydrogen bonds involving the sulfonamide function to form an infinite two-dimensional network parallel to (001).In the crystal of NAQRIJ, dimers are formed through complementary sets of inversion-related O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, which are further connected into zigzag chains by pairwise C\u2014H\u22efN inter\u00adactions that also form inversion dimers. In KUGLIK, the heterocyclic amines are alternately connected by hydrogen bonds thus forming syndiotactic polymeric chains. The hydrogen-bonding network of water mol\u00adecules forms planes parallel to (100). In the crystal of DUSVIZ, mol\u00adecules are linked H)-thione (10\u2005mmol), N-(phen\u00adyl)-2-chloro\u00adacetamide (10\u2005mmol) and sodium acetate trihydrate in ethanol (100\u2005ml) was heated under reflux for 1\u2005h. The reaction mixture was allowed to stand at room temperature overnight. The precipitate that formed was collected and recrystallized from ethanol giving colourless crystals of the title compound, m.p.: 508\u2013510\u2005K, yield 84%. Its IR spectrum showed characteristic absorption bands at 3474\u2005cm\u22121 (OH); 3311\u2005cm\u22121 (NH); 3023\u2005cm\u22121 (C-H aromatic); 2910, 2956\u2005cm\u22121 ; 1800, 1900\u2005cm\u22121 (overtones of phenyl group); 2220\u2005cm\u22121 (C\u2261N) and 1694\u2005cm\u22121 (C=O). Its 1H NMR spectrum exhibited the following signals: \u03b4 10.21 , 7.48\u20137.49 ; 7.22\u20137.25 ; 6.97\u20137.00 ; 6.89\u20136.91 ; 6.75\u20136.77 ; 4.84 ; 4.41\u20134.43 ; 4.04\u20134.11 ; 3.66 ; 3.20\u20133.24 ; 2.83\u20132.85 ; 2.81\u20132.84 ; 2.08 ; 1.86 and 1.21 .A mixture of 7-acetyl-4-cyano-1,6-dimethyl-6-hy\u00addroxy-8-(4-meth\u00adoxy\u00adphen\u00adyl)-5,6,7,8-tetra\u00adhydro\u00adiso\u00adquinoline-3 I, global. DOI: 10.1107/S2056989021005430/yk2151Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021005430/yk2151Isup3.cmlSupporting information file. DOI: 2085564CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Three crystal structures from the reaction of 2,6-di\u00admethyl\u00adphenyl isocyanide with bis\u00ad(anthracene)ferrate(\u22121) are presented. 2]1+ (thf is tetra\u00adhydro\u00adfuran) salt of bis\u00ad(anthracene)ferrate(\u22121), or [Fe(C14H10)2]\u2212, with 2,6-di\u00admethyl\u00adphenyl isocyan\u00adide (CNX\u00adyl) in thf resulted in the formation of two new iron isocyanide complexes, namely, [-anthracene]tris\u00adiron, [Fe(C14H10)(C9H9N)3] or [Fe(CNX\u00adyl)3], and {5,6-bis\u00ad-3--1,2,7-tris\u00ad[imino]-3-azoniahept-3-ene-1,4,7-triido}tris\u00adiron tetra\u00adhydro\u00adfuran disolvate, [Fe(C54H56N6)(C9H9N)3]\u00b72C4H8O or [Fe(C54H56N6)(CNX\u00adyl)3]\u00b72C4H8O, which were characterized by single-crystal X-ray diffraction. The former is likely an inter\u00admediate along the path to the known homoleptic [Fe(CNX\u00adyl)5], while the latter contains a tridentate ligand that is formed from the \u2018coupling\u2019 of six CNXyl ligands. A third crystal structure from this reaction, (7-methyl\u00adindol-1-ido-\u03baN)potassium, [K(C9H8N)(C12H24O6)] or [K(C9H8N)(18-crown-6)], contains a 7-methyl\u00adindol-1-ide anion, in which one CNXyl ligand has shed a proton during its reductive cyclization.The reaction of the [K(18-crown-6)(thf) On this basis, the corresponding reaction with CNXyl in tetra\u00adhydro\u00adfuran, thf, was examined to determine whether the unknown [Fe2(CNX\u00adyl)8]2\u2212 could be accessed. Bis(anthracene)ferrate(\u22121) was also reacted with excess CNXyl in the presence of one equivalent of a reducing agent to see whether the previously reported monometallic [Fe(CNX\u00adyl)4]2\u2212 5] cobaltate(\u22121) has been shown to be an excellent source of spin-paired atomic Co(\u22121) anions in substitution reactions in which both anthracene ferrate(\u22121) with excess CNXyl in THF. First an aliquot was taken from the reaction mixture early on and placed in a 243\u2005K freezer until orange blocks were observed. A single crystal X-ray diffraction experiment revealed these to be [Fe(C14H10)(CNX\u00adyl)3] 1 iron(0). However, the related carbonyl, [Fe(C14H10)(CO)3], has been known for more than 50 years (CNX\u00adyl)3] 2 6X]+ (many variations on R and X) by Zn in the presence of water generated a bis\u00adacetyl\u00adene ligand with protonated nitro\u00adgen atoms (C9H8N)] 3 ]. The hydrogen atom lost during the reduction and cyclization that forms e.g., four), intra\u00adctable tars resulted. It should be noted, however, that this equation is only speculative and requires further investigation for confirmation.Inter\u00adestingly, in support of this equation, when less than eight equivalents of CNXyl were employed (Equation (1)2][Fe(C14H10)2] + 8 CNXyl \u2192 0.5 [Fe(CNX\u00adyl)5] + 0.5 [Fe(C54H56N6)(CNX\u00adyl)3] + [K(18-crown-6)(C9H8N)] portion of a trinuclear mol\u00adecule, 97.5\u00b0 3], 97.7\u00b0 3], 97.9\u00b0 bond lengths \u03c0\u2013\u03c0 inter\u00adactions \u03c0\u2013\u03c0 inter\u00adaction between phenyl rings C56\u2013C61 and C65\u2013C70 [Fe(C14H10)2] and CNXyl were prepared according to previously reported procedures 2] in thf was added CNXyl in thf . The reaction mixture was warmed slowly to room temperature. A solution IR spectrum showed no anionic species, but a broad peak with shoulders that matched the well-known [Fe(CNX\u00adyl)5] were identified as 2 by X-ray diffraction. IR spectroscopy on the crystals (Nujol mull) gave \u03bdCN = 2110w and 2055vs cm\u22121. The filter cake was redissolved in THF and layered with pentane, which resulted in crystals of 3 as determined by a single crystal X-ray experiment. No characterization beyond what is presented above was performed.To a deep-orange solution of . The 18-crown-6 macrocycle is also disordered in a similarly lopsided component ratio; the eight largest residual peaks are the two peaks near the K atom and those for six O atoms of the minor component of disorder. However, the data-to-parameter ratio drops below eight if this disorder is modeled. Thus only the anion disorder was modeled.In 2, bond lengths were restrained toward ideal values and anisotropic displacement parameters were additionally restrained toward the expected motion relative to bond direction.To model the various disordered species, analogous bond lengths and angles were restrained to be similar and anisotropic displacement parameters for proximal atoms were restrained to be similar. For the THF solvent mol\u00adecules in 1 were refined freely to confirm their nature and better describe their true positions. In 2, H7 was also refined freely. All other H atoms were placed geometrically and treated as riding atoms. For 1 and 3 (173\u2005K), methyl\u00adene, C\u2014H = 0.99\u2005\u00c5, aromatic/sp2, C\u2014H = 0.95\u2005\u00c5, with Uiso(H) = 1.2Ueq(C), and methyl, C\u2014H = 0.98\u2005\u00c5, with Uiso(H) = 1.5Ueq(C). For 2 (293\u2005K), methyl\u00adene, C\u2014H = 0.97\u2005\u00c5, aromatic/sp2, C\u2014H = 0.93\u2005\u00c5, N\u2014H = 0.86\u2005\u00c5, with Uiso(H) = 1.2Ueq(C), and methyl, C\u2014H = 0.96\u2005\u00c5, with Uiso(H) = 1.5Ueq(C).The H atoms on the metal-coordinating carbon atoms (C1\u2013C4) of 1 the maximum residual peak of 0.36\u2005e\u2212 \u00c5\u22123 and the deepest hole of \u22120.35\u2005e\u2212 \u00c5\u22123 are found 0.97 and 0.53\u2005\u00c5 from atoms C2 and Fe1, respectively.For 2 the maximum residual peak of 0.38\u2005e\u2212 \u00c5\u22123 and the deepest hole of \u22120.18\u2005e\u2212 \u00c5\u22123 are found 0.81 and 0.39\u2005\u00c5 from atoms H15 and C14, respectively.For 3 the maximum residual peak of 0.58\u2005e\u2212 \u00c5\u22123 and the deepest hole of \u22120.23\u2005e\u2212 \u00c5\u22123 are found 1.15 and 1.25\u2005\u00c5 from atoms C15 and K1, respectively. The peak is part of the minor component of disorder of the 18-crown-6 ring, which was not modeled (see above).For 10.1107/S205698902101313X/yz2014sup1.cifCrystal structure: contains datablock(s) 1, 2, 3, global. DOI: 10.1107/S205698902101313X/yz20141sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S205698902101313X/yz20142sup3.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S205698902101313X/yz20143sup4.hklStructure factors: contains datablock(s) 3. DOI: 2127598, 2127597, 2127596CCDC references: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-021-85972-2, published online 22 March 2021Correction to: The original version of this Article contained an error in Figure\u00a03 where the labels in panel D were incorrect. As a result,\u201cmg/ml\u201dnow reads:\u201cmg/kg\u201dAdditionally,\u201c\u03bcg/ml\u201dnow reads:\u201cmg/kg\u201dFurthermore, in the legend of Figure 3,A) 20\u00a0\u03bcm; in (B) 50\u00a0\u03bcm\u201d\u201cScale bar in (now reads:B) 20\u00a0\u03bcm; in (D) 50\u00a0\u03bcm\u201d\u201cScale bar in (The original Figure"} +{"text": "Intracerebral hemorrhage (ICH) can induce intensively oxidative stress, neuroinflammation, and brain cell apoptosis. However, currently, there is no highly effective treatment available. Puerarin (PUE) possesses excellent neuroprotective effects by suppressing the NF\u2010\u03baB pathway and activating the PI3K/Akt signal, but its role and related mechanisms in ICH\u2010induced early brain injury (EBI) remain unclear. In this study, we intended to observe the effects of PUE and molecular mechanisms on ICH\u2010induced EBI. ICH was induced in rats by collagenase IV injection. PUE was intraperitoneally administrated alone or with simultaneously intracerebroventricular injection of LY294002 . Neurological deficiency, histological impairment, brain edema, hematoma volume, blood\u2013brain barrier destruction, and brain cell apoptosis were evaluated. Western blot, immunohistochemistry staining, reactive oxygen species (ROS) measurement, and enzyme\u2010linked immunosorbent assay were performed. PUE administration at 50\u00a0mg/kg and 100\u00a0mg/kg could significantly reduce ICH\u2010induced neurological deficits and EBI. Moreover, PUE could notably restrain ICH\u2010induced upregulation of the NF\u2010\u03baB pathway, pro\u2010inflammatory cytokines, ROS level, and apoptotic pathway and activate the PI3K/Akt signal. However, LY294002 delivery could efficaciously weaken these neuroprotective effects of PUE. Overall, PUE could attenuate ICH\u2010induced behavioral defects and EBI possibly by PI3K/Akt signal stimulation\u2010mediated inhibition of the NF\u2010\u03baB pathway, and this made PUE a potential candidate as a promising therapeutic option for ICH\u2010induced EBI. The research protocol and animal care were authorized by the Southern Medical University Ethics Committee and were performed according to the National Institutes of Health guidelines on care and use of animals.2.2Supplementary materials, respectively solution of PUE (100\u00a0mg/ml) was prepared. The rats were injected intraperitoneally with either PUE at 50\u00a0mg/kg and 100\u00a0mg/kg, or just DMSO solution 30\u00a0min before modeling and at the time points of 30\u00a0min, 6\u00a0h, 12\u00a0h, 24\u00a0h, and 48\u00a0h after ICH. We selected the dosages and administration route based on previous studies, and intraperitoneal injection of PUE at 50\u00a0mg/kg and 100\u00a0mg/kg could both produce significantly neuroprotective effects.2.5The modified neurological severity score (mNSS) scale was applied to estimate the neurobehavioral deficiency at 24\u00a0h and 72\u00a0h following ICH, which was executed by two experienced researchers who were blinded to animal groups.2.6Paraffin sections of rat brains were made, as previously described.2.7H&E staining was performed as our previous method.2.8V\u00a0=\u00a0T1*S1\u00a0+\u00a0T2*S2 + \u2026 + Tn*Sn .Hematoma volume was assessed as previously reported with some modifications.2.9M1\u2212M2)]/M1*100% .Brain water content (BWC) was measured by the wet/dry weight method, as previously described.2.10Evans blue (EB) dye was applied to assess the disruption of the blood\u2013brain barrier (BBB), as reported previously.2.11+ cells were counted in a blinded manner.At 24\u00a0h and 72\u00a0h after ICH, the brain samples were obtained and paraffin sections were prepared. TUNEL staining was performed with an In Situ Cell Death Detection Kit as our previous method.2.122O2 for 10\u00a0min and antigen blocking with 5% bovine serum albumin for 20\u00a0min, the samples underwent overnighted co\u2010incubation at 4\u00b0C with the following primary antibodies: cleaved caspase\u20103 , Ser536p\u2010NF\u2010\u03baB p65 , NF\u2010\u03baB p65 , 3\u2010nitrotyrosine (3\u2010NT) , and 8\u2010hydroxyguanosine (8\u2010OHdG) . Then, the brain slices were orderly incubated with corresponding secondary antibodies and horseradish peroxidase\u2013streptavidin for 20\u00a0min. After further reaction with 3, 3\u2010diaminobenzidine and being counterstained with hematoxylin, representative IHC images were obtained using a microscope .IHC staining was performed as previously reported.2.13Ser536p\u2010NF\u2010\u03baB p65 (Cat. No.: #3033), NF\u2010\u03baB p65 (Cat. No.: #8242), Ser473p\u2010Akt (Cat. No.: #4060), Akt (Cat. No.: #9272), Bax (Cat. No.: #2772), cleaved caspase\u20103 (Cat. No.: #9664) , Bcl\u20102 , Tyr607p\u2010PI3K , and PI3K . \u03b2\u2010actin and Lamin A were employed as the internal reference. WB protein bands were quantified by ImageJ software . Protein expression levels were indicated by the ratio of interest protein bands to that of \u03b2\u2010actin or Lamin A bands.Western blot (WB) was carried out as our previous method.2.14At 24\u00a0h after ICH, the brain samples were collected and used to detect pro\u2010inflammatory cytokine levels using a Rat TNF\u2010\u03b1 ELISA Kit (Cat. No.: SEA133Ra), Rat IL\u20101\u03b2 ELISA Kit (Cat. No.: SEA563Ra), and Rat IL\u20106 ELISA Kit (Cat. No.: SEA079Ra) as our previous method.2.15The reactive oxygen species (ROS) level was detected using a ROS assay kit at 24\u00a0h after ICH.2.16P\u2010value was statistically significant when <0.05.All data were expressed as means\u00a0\u00b1\u00a0standard deviation (SD). Data analyses were conducted with SPSS 19.0 , and related diagrams were prepared with GraphPad Prism 5 . All data were analyzed using the Shapiro\u2013Wilk and Levene tests. If data satisfy normal distribution and homogeneity of variance, one\u2010way analysis of variance (ANOVA) was considered, and then the least significant difference (LSD) test was applied to compare the difference among multiple experimental groups; conversely, for unsatisfied data, Dunnett's T3 test was adopted. The 33.1P\u00a0<\u00a0.001) and 72\u00a0h (P\u00a0<\u00a0.001) and 100\u00a0mg/kg , yet the beneficial effects of two treatment dosages were not significantly different . The schedule of research, including ICH induction of rats, treatments with different agents , and corresponding experimental assessments, is shown Fig. . The mNS) Figure\u00a0 after IC) Figure\u00a0. Analogo) Figure\u00a0 and 72\u00a0h) Figure\u00a0 followin3.2P\u00a0<\u00a0.001, 24\u00a0h; P\u00a0<\u00a0.01, 72\u00a0h) and 100\u00a0mg/kg could significantly reduce an ICH\u2010induced increase in BWC . Besides, PUE (100\u00a0mg/kg) also could notably repress ICH\u2010induced EB dye extravasation Figure\u00a0. Extrava) Figure\u00a0, and PUE) Figure\u00a0. Similar) Figure\u00a0.3.3P\u00a0<.001, 24\u00a0h and 72\u00a0h) and 100\u00a0mg/kg could markedly reduce the ICH\u2010induced increase in TUNEL+ cells. However, the effects of PUE at 50\u00a0mg/kg and 100\u00a0mg/kg had no statistical difference after ICH Figure\u00a0. Treatme) Figure\u00a0. Typical) Figure\u00a0. PUE cou) Figure\u00a0. BesidesH Figure\u00a0. Consist) Figure\u00a0.3.4P\u00a0<\u00a0.001), p\u2010NF\u2010\u03baB p65 (P\u00a0<\u00a0.05), and nuclear NF\u2010\u03baB p65 (P\u00a0<\u00a0.05) Figure\u00a0,E,G,H, a) Figure\u00a0 at 24\u00a0h ) Figure\u00a0. Similar) Figure\u00a0, p\u2010NF\u2010\u03baB) Figure\u00a0, and nuc) Figure\u00a0 and sign) Figure\u00a0.3.5P\u00a0<\u00a0.001) at 24\u00a0h after ICH at 24\u00a0h following ICH as well could significantly upregulate the expression levels of PI3K Figure\u00a0 and p\u2010AkH Figure\u00a0. Similar) Figure\u00a0 and p\u2010Akl Figure\u00a0. However) Figure\u00a0.3.6P\u00a0<.001) and 100\u00a0mg/kg (P\u00a0<\u00a0.001) could significantly increase the expression level of Bcl\u20102 at 24\u00a0h after ICH and that of cleaved caspase\u20103 at 24\u00a0h following ICH are shown Figure\u00a0. The resH Figure\u00a0. MoreoveH Figure\u00a0. TypicalH Figure\u00a0.3.7+ and 8\u2010OHdG+ cells are shown Figure\u00a0 and 8\u2010OH) Figure\u00a0. Moreove) Figure\u00a0, and PUE) Figure\u00a0 and 100\u00a0) Figure\u00a0, yet the) Figure\u00a0.3.8P\u00a0<\u00a0.001) notably increased Figure\u00a0,H, and P) Figure\u00a0, IL\u20101\u03b2 Figure\u00a0, and IL\u2010) Figure\u00a0. Similar) Figure\u00a0, IL\u20101\u03b2 Figure\u00a0, and IL\u2010) Figure\u00a0 were all) Figure\u00a0,H.3.9P\u00a0<\u00a0.05) was used to ulteriorly explore the protective mechanisms of PUE in ICH\u2010induced EBI, and the acquired results indicated that LY294002 could markedly weaken the beneficial effects of PUE on ICH\u2010induced behavioral deficiency indicated by increasing the mNSS scores Figure\u00a0 and BBB ) Figure\u00a0 at 24\u00a0h 3.10P\u00a0<\u00a0.05) Figure\u00a0, and p\u2010A) Figure\u00a0 proteins) Figure\u00a0 and p\u2010NF) Figure\u00a0 at 24\u00a0h 4Increasing evidence has shown that OS and neuroinflammation are incredibly crucial for ICH\u2010induced EBI, which is characterized by massive brain cell apoptosis.Research studies have confirmed that activation of NFthe \u2010\u03baB signal pathway widely exists in various kinds of CNS damages, including ischemic stroke,Stimulation of the PI3K/Akt signal has also been reported by different groups that it could notably mitigate acute CNS disorder\u2010induced brain injury, including ICH 21H20O9], chemical structure \u20107\u2010hydroxy\u20103\u2010(4\u2010hydroxyphenyl)\u20104H\u20101\u2010benzopyran\u20104\u2010one, Ce Figure\u00a0, belongsPuerarin is also well\u2010confirmed to produce various beneficial effects by activation of the PI3K/Akt signal pathway. A recent report has exhibited that PUE could ameliorate OS\u2010induced neurodegeneration by stimulating the PI3K/Akt signal after TBI, and the specific inhibitor of the PI3K/Akt signaling pathway LY294002 could markedly reduce PUE\u2019s protective effects.Research studies have suggested that the PI3K/Akt signal is the powerful upstream regulator of the NF\u2010\u03baB pathway.Some potential limitations deserve special attention in our study. First, PUE can generate multiple beneficial effects by manipulation of different signal pathways, but we primarily focused on PI3K/Akt signal activation\u2010mediated suppression of the NF\u2010\u03baB pathway. Hence, PUE may exert neuroprotective effects by other signal pathways such as the Nrf2 signaling pathway Overall, our results have indicated that PUE could notably improve ICH\u2010induced EBI and neurological deficiency, and related mechanisms might be involved in the suppression of NF\u2010\u03baB signal pathway activation\u2010induced brain injury partly by the triggering of PI3K/Akt signal pathway\u2010mediated neuroprotection. Our findings might provide a promising therapeutic selection for ICH\u2010induced EBI.All authors listed in this manuscript declared that no any conflicts of interest existed.Jun Zeng: Conceptualization ; Data curation ; Formal analysis ; Investigation ; Methodology ; Software ; Validation ; Visualization ; Writing\u2010original draft . Shizhong Zheng: Conceptualization ; Data curation ; Formal analysis ; Investigation ; Methodology ; Software ; Validation ; Visualization ; Writing\u2010original draft . Yizhao Chen: Conceptualization ; Data curation ; Funding acquisition (lead); Project administration (lead); Resources (lead); Supervision (lead); Writing\u2010review & editing (lead). Yaoming Qu: Formal analysis (supporting); Investigation (supporting); Methodology (supporting); Software (supporting); Visualization (supporting); Writing\u2010review & editing (supporting). Jiayu Xie: Formal analysis (supporting); Investigation (supporting); Methodology (supporting); Software (supporting); Visualization (supporting); Writing\u2010review & editing (supporting). Enhui Hong: Formal analysis (supporting); Investigation (supporting); Methodology (supporting); Software (supporting); Visualization (supporting); Writing\u2010review & editing (supporting). Hongzhu Lv: Formal analysis (supporting); Investigation (supporting); Methodology (supporting); Software (supporting); Visualization (supporting); Writing\u2010review & editing (supporting). Rui Ding: Formal analysis (supporting); Investigation (supporting); Methodology (supporting); Software (supporting); Visualization (supporting); Writing\u2010review & editing (supporting). Liang Feng: Formal analysis (supporting); Investigation (supporting); Methodology (supporting); Software (supporting); Visualization (supporting); Writing\u2010review & editing (supporting). Zhichong Xie: Formal analysis (supporting); Investigation (supporting); Methodology (supporting); Software (supporting); Visualization (supporting); Writing\u2010review & editing (supporting).Fig. S1.Click here for additional data file.Fig. S2.Click here for additional data file.Fig. S3.Click here for additional data file.Fig. S4.Click here for additional data file.Fig. S5.Click here for additional data file.Fig. S6.Click here for additional data file.Supplementary MaterialClick here for additional data file."} +{"text": "II atom has a distorted square-pyramidal coordination environment. The mol\u00adecular structure exhibits an intra\u00admolecular O\u2014H\u22efN hydrogen bond. In the crystal, the mol\u00adecules are linked by inter\u00admolecular C\u2014H\u22efBr hydrogen bonds, generating ribbon structures. These ribbons are linked though inter\u00admolecular C\u2014H\u22efBr hydrogen bonds, forming a two-dimensional network sheet.In the title compound, the Zn 2(C22H19ClN4O)], the ZnII atom adopts a distorted square-pyramidal coordination geometry, formed by two bromido ligands and three N atoms of the bis\u00ad(pyridin-2-ylmeth\u00adyl)amine moiety in the penta\u00addentate ligand containing quinolinol. The ZnII atom is located well above the mean basal plane of the square-based pyramid. The apical position is occupied by a Br atom. The O and N atoms of the quinolinol moiety in the ligand are not coordinated to the ZnII atom. An intra\u00admolecular O\u2014H\u22efN hydrogen bond, generating an S(5) ring motif, stabilizes the mol\u00adecular structure. In the crystal, the mol\u00adecules are linked by inter\u00admolecular C\u2014H\u22efBr hydrogen bonds, generating ribbon structures containing alternating R22(22) and R22(14) rings. These ribbons are linked through an inter\u00admolecular C\u2014H\u22efBr hydrogen bond, forming a two-dimensional network sheet.In the title compound, [ZnBr Its derivatives are utilized as chemosensors for detecting ZnII at low concentration in biological samples bromide. Here, the crystal structure of the respective title compound is reported.8-Quinolinol (Hq) is a notable bidentate ligand and an excellent analytical reagent for the determination of the concentration and separation of metal ions and three N atoms of the dpa moiety in HClqdpa forming the ZnBr2(dpa) unit. The Hq moiety of the penta\u00addentate ligand (HClqdpa) is not coordinated to the ZnII center. The five-coordinate geometry parameter, \u03c4 = (\u03b2\u00a0\u2212\u00a0\u03b1)/60, derived from the two largest angles (\u03b1 < \u03b2) in a structure has ideal values of 0 for square-pyramidal and of 1 for trigonal\u2013bipyramidal geometry \u2005\u00c5 above the mean basal plane (Br2/N8/N7/N9) of the square-based pyramid. The dpa moiety is meridionally bound to the ZnII atom. The apical position is occupied by the Br1 atom with the apical bond being slightly elongated to 2.4419\u2005(4)\u2005\u00c5 compared to the equatorial Br2\u2014Zn3 bond length of 2.4085\u2005(4)\u2005\u00c5. The Zn\u2014N bond lengths in the title compound are 2.1455\u2005(18) and 2.1497\u2005(18)\u2005\u00c5 for the pyridyl atoms , and 2.2670\u2005(18)\u2005\u00c5 for the tertiary atom N7. In comparison, the Zn\u2014N bond lengths in the crystal structure of a related complex with a mesityl methyl\u00adene-appended dpa derivative are 2.093\u2005(3), 2.066\u2005(3), and 2.521\u2005(3)\u2005\u00c5 ring motif 1\u00a0\u2212\u00a0x, \u2212y, \u2212z] 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z] x\u00a0+\u00a01, y\u00a0\u2212\u00a01, z] (Table\u00a01C(6) chain motif running along Table\u00a01, forming] Table\u00a01, which ffs Fig.\u00a02. In the ] Table\u00a01, forming0] Fig.\u00a03. The rib] Fig.\u00a03.et al., 2016ConQuest amino]\u00admethyl fragment as ligand gave 517 hits, and among those, eight hits with two bromido ligands. Of these eight analogues, three structures are complexes with dpa bearing a tertiary N donor atom directly bound to an aromatic moiety , the tertiary N atoms are bound to aliphatic carbon atoms as in the title complex. Four of these five closely related structures exhibit square-pyramidal geometries with dpa being meridionally coordinated meth\u00adyl]propan-2-amine}\u00addibromido\u00adzinc(II) . Of these, six structures (three compounds) are ion-pairs between tetra\u00adchlorido\u00adzincate(II) and an 8-hy\u00addroxy\u00adquinolin-1-ium (H2q+) derivative, for example, (H2q)2[ZnCl4] and quinolin-8-lato derivatives, e.g. 8-hy\u00addroxy-2-methyl\u00adquinolino\u00adlinium di\u00adiodo\u00ad(2-methyl\u00aduinolin-8-lato)zinc(II) tris\u00ad(4-nitro\u00adphenol) bis\u00adzinc(II) dihydrate zinc(II) trihydrate amino]\u00admethyl at the 2-position of Hq or respective derivatives, gave three hits was dissolved in 15\u2005mL of hot aceto\u00adnitrile. Then a solution of zinc(II) bromide in 15\u2005mL of hot aceto\u00adnitrile was added to the ligand solution. The mixture was stirred for 20\u2005min at 333\u2005K. After removal of the solvent at room temperature in air for one week, colorless crystals of the title compound were obtained . Analysis calculated for CUiso(H) = 1.2Ueq(C). One outlier reflex (002) was omitted from the refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989022001530/yz2016sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989022001530/yz2016Isup2.hklStructure factors: contains datablock(s) I. DOI: 2150991CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Heterodera microulae sp. n., was isolated from the roots and rhizosphere soil of Microula sikkimensis in China. Morphologically, the new species is characterized by lemon-shaped body with an extruded neck and obtuse vulval cone. The vulval cone of the new species appeared to be ambifenestrate without bullae and a weak underbridge. The second-stage juveniles have a longer body length with four lateral lines, strong stylets with rounded and flat stylet knobs, tail with a comparatively longer hyaline area, and a sharp terminus. The phylogenetic analyses based on ITS-rDNA, D2-D3 of 28S rDNA, and COI sequences revealed that the new species formed a separate clade from other Heterodera species in Goettingiana group, which further support the unique status of H.\u00a0microulae sp. n. Therefore, it is described herein as a new species of genus Heterodera; additionally, the present study provided the first record of Goettingiana group in Gansu Province, China.A new cyst-forming nematode, Heterodera was erected by Schmidt (1871) and currently contains about 80 species units are good candidate genes for molecular taxonomic and phylogenetic studies were recovered from hatched eggs and kept in water suspension until further use and AB28 (5\u2032-ATATGCTTAAGTTCAGCGGGT-3\u2032) (COI gene was amplified using primers Het-coxiF (5\u2032-TAGTTGATCGTAATTTTAATGG-3\u2032) and Het-coxiR (5\u2032-CCTAAAACATAATGAAAATGWGC-3\u2032) . Three sets of primers were used in the PCR analyses to amplify sequences of the ITS, D2-D3 expansion segments of 28S, and GGGT-3\u2032) . The 28SGGGT-3\u2032) . FinallyGWGC-3\u2032) . PCR conCOI gene) were compared with known sequences of Heterodera using BLASTn homology search program. Outgroup taxa for phylogenetic analyses were selected based on the previously published studies (http://tree.bio.ed.ac.uk/software/figtree/) . The speHeterodera microulae sp. n. It is lemon-shaped with an obtuse vulval cone, neck extruding, and cuticle thick with an irregular zig-zag pattern. The color was white to pale to medium brown; remnants of the subcrystalline layer were rarely present. The egg sac was usually absent , 3B, C. The female was lemon-shaped, pearl white, or pale yellow in color. It was rarely rounded with a protruding neck and vulva, the subcrystalline layer was present, and the egg sac absent . There wThe body was straight or slightly curved ventrally after heat treatment . The lipBody hyaline without any markings was presented; juveniles folded six times .The male was not found.Holotype and paratype material were deposited in the nematode collection of the Department of Plant Protection, Biocontrol Engineering Laboratory of Crop Diseases and Pests of Gansu Province, Lanzhou, China.Heterodera microulae sp. n. was collected from the roots and rhizosphere soil of Microula sikkimensis Hemsl. in Tianzhu county of Gansu Province, China. The geographical position is N 37\u00b011\u203246\u2033; E 102\u00b047\u20326\u2033. This site was located in continental highland with the vegetation type of meadow grassland and the soil is composed of chernozems. The climatic parameters of the site include 450\u2009mm of average rainfall and an approximate \u22122 air temperature.The species is named after the host of its isolation.Heterodera microulae sp. n. is characterized by having lemon-shaped cysts that have protruding necks and obtuse vulval cones. The cysts are 414 to 543-\u00b5m long and 305 to 456-\u00b5m wide having ambifenestrate vulval cone and bullae are absent. Females are white in color with a subcrystalline layer. Second-stage juveniles are straight or slightly curved ventrally with four incisures in the lateral field. The juveniles are 506 to 628-\u00b5m long having strong stylets with well-developed rounded stylet knobs, genital primordium situated at 59 to 62% of body length, and tail 49 to 61-\u00b5m long with a hyaline portion of 24 to 33\u2009\u00b5m. Eggs are hyaline without any markings; juveniles inside the eggs form sixfold.Goettingiana group of Heterodera; currently, the group contains seven valid species, viz, Heterodera goettingiana , absence of bullae (vs few), weak underbridge (vs 117\u2009\u00b5m), longer J2s body length (568\u2009\u00b5m vs 486\u2009\u00b5m), stylet knobs rounded and flat or slightly concave anteriorly vs smoothly rounded to slightly hook-shaped with a recurved anterior surface, longer distance of median bulb from the anterior end (MB) (86\u2009\u00b5m vs 70\u2009\u00b5m), shorter excretory pore distance from the anterior end (114\u2009\u00b5m vs 158\u2009\u00b5m), and shorter length of hyaline tail portion (29\u2009\u00b5m vs 37\u2009\u00b5m).The new species differs from H. carotae by having a bigger size of cysts (495\u2009\u00d7\u2009384\u2009\u00b5m vs 408\u2009\u00d7\u2009309\u2009\u00b5m), shorter vulval slit length (39\u2009\u00b5m vs 47\u2009\u00b5m), longer J2s body length (568\u2009\u00b5m vs 422\u2009\u00b5m), stylet knobs rounded and flat or slightly concave anteriorly vs concave anterior face, higher MB value (86\u2009\u00b5m vs 66\u2009\u00b5m), longer excretory pore distance from the anterior end (114\u2009\u00b5m vs 99\u2009\u00b5m), and longer tail length (57\u2009\u00b5m vs 52\u2009\u00b5m).The new species is differentiated from H. cruciferae by having a bigger size of cysts (495\u2009\u00d7\u2009384\u2009\u00b5m vs 429\u2009\u00d7\u2009333\u2009\u00b5m), slightly shorter fenestral length (31\u2009\u00b5m vs 34\u2009\u00b5m), shorter vulval length (39\u2009\u00b5m vs 45\u2009\u00b5m), longer J2s body length (568\u2009\u00b5m vs 431\u2009\u00b5m), higher MB value (86\u2009\u00b5m vs 68\u2009\u00b5m), longer excretory pore distance from the anterior end (114\u2009\u00b5m vs 101\u2009\u00b5m), longer tail length (57\u2009\u00b5m vs 50\u2009\u00b5m), and longer length of hyaline tail portion (29\u2009\u00b5m vs 25\u2009\u00b5m).The new species differs from H. persica by a shorter fenestral length (31\u2009\u00b5m vs 47\u2009\u00b5m), absence of bullae (vs present), shorter vulval slit length (39\u2009\u00b5m vs 49\u2009\u00b5m), longer J2s body length (568\u2009\u00b5m vs 440\u2009\u00b5m), stylet knobs , longer stylet (26\u2009\u00b5m vs 23\u2009\u00b5m), higher MB value (86\u2009\u00b5m vs 70\u2009\u00b5m), longer excretory pore distance from the anterior end (114\u2009\u00b5m vs 103\u2009\u00b5m), longer tail length (57\u2009\u00b5m vs 47\u2009\u00b5m), and longer length of hyaline tail portion (29\u2009\u00b5m vs 24\u2009\u00b5m).The new species differs from H. urticae, the new species has a smaller size of cysts (495\u2009\u00d7\u2009384\u2009\u00b5m vs 492\u2009\u00d7\u2009435\u2009\u00b5m), vulval cone obtrusive (vs unobtrusive) and absence of egg sac (vs presence), shorter fenestral length (31\u2009\u00b5m vs 38\u2009\u00b5m), shorter vulval slit length (39\u2009\u00b5m vs 46\u2009\u00b5m), longer J2s body length (568\u2009\u00b5m vs 541\u2009\u00b5m), shorter DGO (8\u2009\u00b5m vs 5\u2009\u00b5m), and shorter excretory pore distance from the anterior end (114\u2009\u00b5m vs 130\u2009\u00b5m).Compared with H. circeae having a smaller size of cysts (495\u2009\u00d7\u2009384\u2009\u00b5m vs 555\u2009\u00d7\u2009397\u2009\u00b5m), a shorter fenestral length (31\u2009\u00b5m vs 43\u2009\u00b5m), vulval slit length (39\u2009\u00b5m vs 48\u2009\u00b5m), longer J2s body length (568\u2009\u00b5m vs 434\u2009\u00b5m), stylet knobs (rounded and slightly sloping posteriorly vs rounded and flat or slightly concave anteriorly), higher MB value (86\u2009\u00b5m vs 70\u2009\u00b5m), longer excretory pore distance from the anterior end (114\u2009\u00b5m vs 101\u2009\u00b5m), longer tail length (57\u2009\u00b5m vs 52\u2009\u00b5m), and longer length of hyaline tail portion (29\u2009\u00b5m vs 26\u2009\u00b5m).The new species differs from H. scutellariae, having smaller cysts (495\u2009\u00d7\u2009384\u2009\u00b5m vs 560\u2009\u00d7\u2009424\u2009\u00b5m), by a shorter fenestral length (31\u2009\u00b5m vs 35\u2009\u00b5m), vulval slit length (39\u2009\u00b5m vs 43\u2009\u00b5m), longer J2s body length (568\u2009\u00b5m vs 408\u2009\u00b5m), higher MB value (86\u2009\u00b5m vs 62\u2009\u00b5m), longer excretory pore distance from the anterior end (114\u2009\u00b5m vs 89\u2009\u00b5m), longer tail length (57\u2009\u00b5m vs 47\u2009\u00b5m), and longer length of hyaline tail portion (29\u2009\u00b5m vs 25\u2009\u00b5m).The new species differs from H. microulae sp. n. with other valid species of Goettingiana group are listed in Additionally, comparative morphological and morphometric characters of H. microulae sp. n. sequences of D2-D3 region of 28\u2009S (734\u2009bp), ITS (993\u2009bp), and COI (415\u2009bp) gene were obtained and submitted to the GenBank.The H. microulae sp. n. showed 97.09% (19-bp difference), 97.66 to 98.49% (11-17-bp difference), 98.38% (9-bp difference), 98.62% (9-bp difference), 98.45% (11-bp difference), and 99.86 to 100% (0-1-bp difference) sequence identities with H. goettingiana (DQ328697), H. carotae (KX463292 and KX463293), H. cruciferae (KP114546), H. urticae (DQ328696), Heterodera sp. RH-2010 (GU456692) from Iran, and Heterodera sp. DP-2010 (HM560856 and HM560855) from Qinghai, China, respectively. The Bayesian phylogenetic tree of the D2-D3 of 28S gene from Qinghai, China and forms a 100% supported clade.The D2-D3 of 28S-rRNA sequence (accession no. MT573436) of 28S gene represenH. microulae sp. n. with other Goettingiana group species is as follows: 0.20% (2-bp difference), 0.4 to 0.5% (4-bp difference), 3.02% (29-bp difference), 5.01% (48-bp difference), 5.11% (49-bp difference), 7.45% (72-bp difference), 6.77 to 6.95% (67-68-bp difference), 6.29 to 7.25% (66-70-bp difference), and 7.41 to 8% (74-77-bp difference) for Heterodera sp. DP-2010 (HM560761), H. goettingiana , H. persica (AF498377), H. scutellariae (AY368995), H. circeae (AY368994), H. urticae (AF274412), H. carotae , H. cruciferae , and H. goettingiana , respectively. The Bayesian phylogenetic tree of the ITS gene clustered with H. persica (AF498377), H. scutellariae (AY368994), H. circeae (AY368995), Heterodera sp. DP-2010 (HM560791), and H. goettingiana from Qinghai, China with high-probability support (pp\u2009=\u200991%). It is also noted that sequences of H.\u00a0goettingiana from Qinghai, China, clustered outside with other H. goettingiana subclades and should be considered a misidentification. However, H. microulae sp. n. (MT573437) is clustered with H. sp. DP-2010 (HM560791) and H. goettingiana from Qinghai, China, with 100% support. It is also noted that H. microulae sp. n. (MT573437) clustered with two Chinese populations of Heterodera species with 100% support.The ITS-rDNA sequence (accession no. MT573437) divergence of ITS gene represenCOI gene sequence of H. microulae sp. n. showed 87.21 to 89.53% (differing from 36 to 44\u2009bp), 88.19% (differing from 43\u2009bp), 88.67 to 88.92% (differing from 46 to 47\u2009bp), and 88.67 to 89.40% (differing from 44 to 47\u2009bp), sequence identities with H. goettingiana (KY129829-KY129831), H. urticae (MK093155 and MK093156), H. cruciferae (MG563230 and MG563234), and H. carotae , respectively. The Bayesian phylogenetic tree of the COI gene and Subbotin et al. (2001) used J2\u2019s lateral field characters and host preferences to separate Heterodera species into different groups . The key morphological characters of the Goettingiana group include lemon-shaped cysts having a protruding neck, ambifenestration, and absence of bullae ; some species may have an egg sac, vulval slit length\u2009>\u200935\u2009\u00b5m, a thin vulval bridge, fenestral length (30-45\u2009\u00b5m), and a weak underbridge. There were second-stage juveniles with body length\u2009>\u2009400\u2009\u00b5m, stylet length\u2009>\u200920\u2009\u00b5m, tail length\u2009>\u200945\u2009\u00b5m, hyaline tail portion\u2009>\u200920\u2009\u00b5m, and lateral field with four lines and H. goettingiana (HM370423 and HM370425) from Qinghai, China, formed a well-supported molecular clade with the H. microulae sp. n. Moreover, the nucleotide differences of these sequences with our new species sequences are also very low (2-4-bp difference for ITS and 0-1\u2009bp for 28S). Previously, H. goettingiana (HM370423 and HM370425) from Qinghai, China, might be a case of misidentification. Based on our phylogenetic and sequence analysis results, we regard Heterodera sp. DP-2010 and H. goettingiana (HM370423 and HM370425) as H. microulae sp. n.Phylogenetically, it is evident that Heterodera microulae sp. n. is isolated from Microula sikkimensis, it is a biennial herbaceous plant that grows in forests, meadows, and forest edges at altitudes of 2,200 to 4,700\u2009m, and it is widely distributed in South and East Asian countries are growing in the same locality. Therefore, host-suitability tests of H. microulae sp. n. are an open research field to investigate the damage potential of this species.The present study described a new species found in the rhizosphere of"} +{"text": "The structure of the racemic form of the diuretic drug trichlorme\u00adthia\u00adzide was determined from laboratory X-ray powder diffraction data: the extended structure features an intricate combination of N\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 and C\u2014Cl\u22ef\u03c0 inter\u00adactions. RS)-trichlorme\u00adthia\u00adzide , C8H8Cl3N3O4S2 (RS-TCMZ), a diuretic drug used in the treatment of oedema and hypertension, was determined from laboratory X-ray powder diffraction data using DASH [David et al. . Energy framework calculations confirm the major contribution of electrostatic inter\u00adactions (Eelec) to the total energy (Etot). A comparison with the structure of S-TCMZ is also presented.The structure of racemic ( al. 2006. J. Applelho 2018. J. Appl H-1\u03bb6,2,4-benzo\u00adthia\u00addi\u00adazine-7-sulfonamide (C8H8Cl3N3O4S2), is a diuretic drug derived from thia\u00adzide, the precursor of a classic family of diuretic compounds, discovered in the 1950s. The first approved drug of this class, chloro\u00adthia\u00adzide, was marketed under the trade name Diuril in 1958 , systematic name 6-chloro-3-(di\u00adchloro\u00admeth\u00adyl)-1,1-dioxo-3,4-di\u00adhydro-2Mogul geometry check exhibits a conformation that could be described as distorted half-chair to distorted envelope at N3 and equatorial conformations. The almost planar benzosulfonamide ring (ring B) makes an angle of 8.2\u2005(2)\u00b0 with the best plane through the thia\u00adzide ring. The mol\u00adecule is oriented almost parallel to the a-axis as indicated by a 3.11\u2005(8)\u00b0 angle \u00b0].The asymmetric unit contains one TCMZ mol\u00adecule Fig.\u00a01: the steS-configuration in racemic TCMZ with the mol\u00adecule of the S-enanti\u00adomer in KIKCUD. When flexibility is allowed in the superposition , the r.m.s.d. deviation is 0.070 and the maximum deviation (max. D) is 0.146\u2005\u00c5. Without flexibility, the values for r.m.s.d. and max. D are 0.785 and 2.763\u2005\u00c5, respectively . The difference between the two conformations lies in the orientation of the sulfonamide group and leads to differences in the hydrogen-bonding patterns between the two compounds as discussed below.Fig.\u00a02on Fig.\u00a02a, the rly Fig.\u00a02b. The dIntra\u00admolecular hydrogen bondsRS-TCMZ \u2005\u00c5 and a D\u2014H\u22efA angle of 106.9\u2005(5)\u00b0. A second intra\u00admolecular hydrogen bond occurs between C7\u2014H7 and O4 . The third contact, N3\u2014H3A\u22efCl3, has geometric parameters 2.767\u2005(8)\u2005\u00c5 and 102.1\u2005(5)\u00b0. The three hydrogen bonds can all be represented by the graph-set symbol S(5) .Three different intra\u00admolecular hydrogen bonds are present in MZ Fig.\u00a03b.Inter\u00admolecular hydrogen bondsRS-TCMZ is very rich and relevant geometric parameters are summarized in Table\u00a01a, an A as the donated H alternate with a motif , occur between mol\u00adecules related by the symmetry operation \u2212x, 2\u00a0\u2212\u00a0y, \u2212z . At the same time, the original mol\u00adecule inter\u00adacts via a C\u2014Cl\u22ef\u03c0 contact of 3.761\u2005(5)\u2005\u00c5 with another mol\u00adecule related by symmetry operation 1\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z . The mol\u00adecules are arranged as head-to-tail dimers producing chains along [101] as depicted in Fig.\u00a05c. In contrast, in the structure of S-TCMZ only C\u2014Cl\u22ef\u03c0 inter\u00adactions are observed (Table\u00a01d = 3.456\u2005(2)\u2005\u00c5] than in RS-TCMZ.In addition, \u03c0\u2013\u03c0 and C\u2014Cl\u22ef\u03c0 inter\u00adactions , which form columns along the b-axis. These columns are further connected by N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds .The structure of ns Fig.\u00a06c, whichds Fig.\u00a06d.et al., 2016S-trichlorme\u00adthia\u00adzide shows that they correspond to the same phase. It is worth mentioning that a broader search of the CSD resulted in 100 structures related to TCMZ, among them chloro\u00adthia\u00adzide and hydro\u00adchloro\u00adthia\u00adzide, their polymorphs, derivatives, solvates, and co-crystals.A search in the Cambridge Structural Database -Trichlorme\u00adthia\u00adzide was kindly provided by Tecnoqu\u00edmicas . Based on the FT\u2013IR spectra and the quality of the preliminary diffraction patterns, the present study was carried out on the sample as it was received. Crystallization experiments in different solvents, in search of possible polymorphs, are underway in our laboratories.\u2005\u00c5, b = 8.8919\u2005(9)\u2005\u00c5, c = 9.720\u2005(2)\u2005\u00c5, \u03b1 = 91.30\u2005(1)\u00b0, \u03b2 = 106.07\u2005(2)\u00b0, \u03b3 = 97.19\u2005(1)\u00b0, V = 693.4\u2005(2)\u2005\u00c53. The de Wolf , respectively. A reduced-cell search in the CSD using GRACE analysis for this mol\u00adecule. The HBP analysis was carried out with supporting information.The HBP tool provides an insight into the expected intra- and inter\u00admolecular hydrogen bonds in the structures. For the analysis, the donor atoms considered were N1 (sulfonamide), N2 (secondary amine) and N3 (next to the sulfonyl group). The acceptors were Cl1 (aryl chloride), Cl2/Cl3 , N2 (secondary amine), O1/O2 (sulfonamide), and O3/O4 (sulfon\u00adyl). The area under the receiver operating characteristics (ROC) curve was 0.863, indicating good statistical discrimination in the analysis. The results of the calculations are presented in the B\u22efCl1 (0.60). This hydrogen bond is observed only in the S-enanti\u00adomer. The intra\u00admolecular inter\u00adaction involving N3\u2014H3A\u22efCl3, observed in the two structures, has the second highest propensity value (0.48).The intra\u00admolecular hydrogen bond with the highest propensity is N1\u2014H1B\u22efO3 and N1\u2014H1A\u22efO4) have the highest propensities (0.69). They are present in the structure of RS-TCMZ (motifs II and III). However, only one of them (N1\u2014H1A\u22efO4) is present in the S-enanti\u00adomer. The next two inter\u00adactions with highest propensities (0.68) are between the H and O atoms of the sulfonamide groups of two neighboring mol\u00adecules. One of them (N1\u2014H1B\u22efO2) is observed only in the S-enanti\u00adomer.Regarding the inter\u00admolecular inter\u00adactions, two hydrogen bonds involving the hydrogen atoms bonded to the nitro\u00adgen of the sulfonamide group and the two oxygen atoms of the sulfonyl group (N1\u2014H1A) and for the secondary amine (N2\u2014H2A) with the sulfonyl O atoms , which are not present in either structure. However, N3\u2014H3A\u22efO1 and N3\u2014H3A\u22efO2 contacts with 0.42 propensities are displayed in S-TCMZ and RS-TCMZ (motifs I and IV), respectively. In addition, the hydrogen bond N2\u2014H2A\u22efO1 is present in RS-TCMZ (motifs V and VI) but not in S-TCMZ. The hydrogen bond N2\u2014H2A\u22efN1 was not predicted because the N1 atom was not considered an acceptor. The hydrogen-bond patterns found in the two structures are consistent with the hydrogen-bond propensity analysis results. Every donor and acceptor in RS-TCMZ and in S-TCMZ has a hydrogen-bond coordination with a high likelihood. Figure S2 of the additional supporting information shows the putative landscape for trichlorme\u00adthia\u00adzide. The two structures fall in the high propensity and hydrogen-bond coordination zone.The CSD statistics predicts hydrogen bonds for the sulfonyl nitro\u00adgen atom with 36.0 and 16.9%, respectively. The next inter\u00adaction, O\u22efCl, is slightly less important in RS-TCMZ than in S-TCMZ (8.7% versus 9.6%). The remaining inter\u00adactions differ in order of importance. For example, the H\u22efH inter\u00adaction is more important (8.5%) in RS-TCMZ than in S-TCMZ (7.2%). It is worth noting that the fingerprint plot delineated into the H\u22efH inter\u00adaction for RS-TCMZ shows a tip at de + di = 2.20\u2005\u00c5, which is less than 2 times the van der Waals radii of hydrogen. In contrast, in S-TCMZ this inter\u00adaction is dispersed over a range of de + di values. Weaker inter\u00adactions such as \u03c0\u2013\u03c0 contacts are present only in racemic TCMZ and they represent 1.8% of the contribution to the Hirshfeld surface. The Cl\u22ef\u03c0 inter\u00adaction is more important in S-TCMZ, contributing 9.1% in contrast to RS-TCMZ where it represents 2.8%. This is the result of two inter\u00adactions in the S-enanti\u00adomer that lead to layers parallel to the ab plane. In RS-TCMZ, the Cl\u22ef\u03c0 inter\u00adactions alternate with \u03c0\u2013\u03c0 contacts to produce chains nearly along [101]. Another inter\u00adesting feature is displayed by the H\u22efN contacts. There is a lower degree of directionality and strength of this inter\u00adaction in RS-TCMZ (2.1%) than in S-TCMZ (3.3%) as a result of the additional N2\u2014H2A\u22efN1 inter\u00adaction in the latter.Fig.\u00a08ts Fig.\u00a08m\u20138x witEele), dispersive (Edis), and total energies (Etot) for the inter\u00admolecular inter\u00adactions in RS-TCMZ and S-TCMZ were calculated with CrystalExplorer21 and dispersive components are similar although their contributions are quite different. They result in an offset tile arrangement for Etot when viewed down the b-axis direction . In the structure of S-TCMZ, Eele and Edis make similar contributions to Etot and their topology is similar . The pattern viewed down the b-axis direction resembles a herringbone arrangement .As depicted in Fig.\u00a09on Fig.\u00a09c. In thar Fig.\u00a09d and 9ent Fig.\u00a09f.supporting information). The stretches for the secondary N\u2014H grouping of the sulfonamide group appear at 3387 and 3322\u2005cm\u22121 followed by the stretching bands of the S\u2014N\u2014H and N\u2014H groups of the amine on the di\u00adhydro\u00adthia\u00addiazine at 3281 and 3232\u2005cm\u22121, respectively. The stretches of the Csp2\u2014H (3150\u20133100\u2005cm\u22121) and Csp3\u2014H (3000\u20132900\u2005cm\u22121) bonds are observed as weak bands. The Csp2\u2014Csp2 stretch of the aromatic ring appears at 1596\u2005cm\u22121 while the C\u2014N and S\u2014N stretches overlap at 1351 and 1332\u2005cm\u22121. The stretches of the two S=O groups appear as strong absorptions at 1176 and 1157\u2005cm\u22121. Table S2 summarizes the assignment of the most important absorptions for RS-TCMZ.The FT\u2013IR spectrum shows the absorption bands expected for TCMZ recorded indicates the material is stable up to 240\u00b0C. A series of weight loss events occur from 240\u00b0C to 450\u00b0C. A 24.2% weight loss (2.270\u2005mg) between 245 and 301\u00b0C coincides with the first two transitions in the DSC . The endotherm at 281.1\u00b0C (\u0394H = 81.19\u2005J\u2005g\u22121) is associated with melting of the material. This transition is followed by an exotherm with peak temperature 287.9\u00b0C (\u0394H = 103.70\u2005J\u2005g\u22121). The TGA curve shows two continuous weight loss processes at 302\u2013384\u00b0C and 384\u2013448\u00b0C , associated with ill-defined transitions in the DSC. The total weight loss due to decomposition is 49.6%.The TGA curve TCMZ, I. DOI: 10.1107/S2056989021013633/hb7999Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021013633/hb7999Isup3.molSupporting information file. DOI: 10.1107/S2056989021013633/hb7999sup4.pdfPowder pattern superposition, Hydrogen bond propensity analysis, Hirshfeld surface, energy frameworks calculations, FT-IR, and TGA/DSC. DOI: Click here for additional data file.10.1107/S2056989021013633/hb7999Isup5.cmlSupporting information file. DOI: 2034096, 2034096CCDC references: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the mol\u00adecules are connected by N\u2014H\u22efO hydrogen-bond pairs along the b-axis direction as dimers with R22(8) and R22(14) ring motifs and as ribbons formed by inter\u00admolecular C\u2014H\u22efN hydrogen bonds. There are weak van der Waals inter\u00adactions between the ribbons. The most important contributions to the surface contacts are from H\u22efH (34.9%), O\u22efH/H\u22efO (19.2%), C\u22efH/H\u22efC (11.9%), Cl\u22efH/H\u22efCl (10.7%) and N\u22efH/H\u22efN (10.4%) inter\u00adactions, as concluded from a Hirshfeld surface analysis.The mol\u00adecular conformation of the title compound, C The planes of the 2,3-di\u00adhydro-1H-indole ring system and the 4H-pyran ring are approximately perpendicular to each other, subtending a dihedral angle of 84.52\u2005(5)\u00b0. The C5\u2014C6\u2014C11\u2014Cl1, C6\u2014C5\u2014C8\u2014O2, C6\u2014C5\u2014C8\u2014O3, C5\u2014C8\u2014O3\u2014C9 and C8\u2014O3\u2014C9\u2014C10 torsion angles are \u2212103.28\u2005(13), \u221229.78\u2005(18), 150.69\u2005(11), 178.03\u2005(10) and \u2212169.29\u2005(12)\u00b0, respectively. An intra\u00admolecular C11\u2014H11B\u22efO2 contact stabilizes the mol\u00adecular conformation of the title compound ring motif inter\u00adactions are the major factor in the crystal packing with O\u22efH/H\u22efO , C\u22efH/H\u22efC , Cl\u22efH/H\u22efCl and N\u22efH/H\u22efN inter\u00adactions representing the next highest contributions. Other weak inter\u00adactions (contribution percent\u00adages) are O\u22efN/N\u22efO (2.3%), O\u22efC/C\u22efO (2.1%), N\u22efC/C\u22efN (2.1%), Cl\u22efN/N\u22efCl (1.7%), Cl\u22efO/O\u22efCl (1.4%), C\u22efC (1.0%), N\u22efN (0.7%), O\u22efO (0.6%), Cl\u22efC/C\u22efCl (0.6%) and Cl\u22efCl (0.3%).Fig.\u00a07et al., 2016H-pyran-3-carbo\u00adnitrile as the main skeleton revealed the presence of three structures, ethyl 6-amino-2-(chloro\u00admeth\u00adyl)-5-cyano-4-(o-tol\u00adyl)-4H-pyran-3-carb\u00adoxyl\u00adate and C(8). Combination of these primary motifs leads to a secondary In the crystal of HIRNUS, the six-membered pyran ring adopts a near-boat conformation. The crystal structure features two intra\u00admolecular C\u2014H\u22efO inter\u00adactions and the crystal packing is stabilized by inter\u00admolecular N\u2014H\u22efO hydrogen bonds. These lead to two primary motifs, In the crystal of JEGWEX, a potential precursor for fluoro\u00adquinoline synthesis, the pyran ring is nearly planar, with the most outlying atoms displaced from the best-plane fit through all non-H atoms by 0.163\u2005(2) and 0.118\u2005(2)\u2005\u00c5. The mol\u00adecules are arranged in layers oriented parallel to the (011) plane. In addition, the mol\u00adecules are linked by a weak C\u2014H\u22efO hydrogen bond, which gives rise to chains with base vector [111].QT = 0.085\u2005(7)\u2005\u00c5, \u03b8 = 84\u2005(5)\u00b0 and \u03c6 = 154\u2005(5)\u00b0. In the crystal, mol\u00adecules are linked as dimers by pairs of N\u2014H\u22efO hydrogen bonds, forming ribbons along the b-axis direction. These ribbons are connected by weak van der Waals inter\u00adactions, stabilizing the mol\u00adecular packing.In WIMBEC, the pyran ring exhibits a near-boat conformation with puckering parameters et al., 2015et al., 2018The title compound was synthesized using previously reported procedures , N2\u2014H2A = 0.843\u2005(19) and N2\u2014H2B = 0.889\u2005(18)\u2005\u00c5], but their isotropic displacement parameters were constrained to take a value of 1.2Ueq(N). All H atoms bound to C atoms were positioned geometrically and refined as riding with C\u2014H = 0.95 (aromatic), 0.99 (methyl\u00adene) and 0.98\u2005\u00c5 (meth\u00adyl), with Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for the others. Four reflections, 0 0 1, 0 1 0, 1 0 0 and 1 2 0, affected by the incident beam-stop and owing to poor agreement between observed and calculated intensities, and five outliers, Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021006459/vm2250sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021006459/vm2250Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021006459/vm2250Isup3.cmlSupporting information file. DOI: 2091350CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular complex is based on the heterobimetallic AgI\u2014VV fragment {AgI2(VVO2F2)2(tr)4} supported by four 1,2,4-triazole ligands [4-benzyl-]. The triazole functional group demonstrates homo- and heterometallic connectivity (Ag\u2014Ag and Ag\u2014V) of the metal centers through the [\u2013NN\u2013] double and single bridges, respectively. The vanadium atom possesses a distorted trigonal\u2013bipyramidal coordination environment [VO2F2N] with the Reedijk structural parameter \u03c4 = 0.59. In the crystal, C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds as well as C\u2014H\u22ef\u03c0 contacts are observed involving the organic ligands and the vanadium oxofluoride anions. A Hirshfeld surface analysis of the hydrogen-bonding inter\u00adactions is also described.The crystal structure of the title compound, [Ag The 1,2,4-triazole heterocycle, as a functional group, demonstrates a favorable coordination affinity towards AgI cations, connecting them into polynuclear units 2(tr)4} SBUs with bi-1,2,4-triazole ligands with different geometries 2(2Phtr-CH)4] (I), which has the ligand 4-benzyl- (tr-CH2Ph).There is considerable inter\u00adest in the chemistry of organic\u2013inorganic hybrids, including the vanadium oxide\u2013fluoride (VOF) matrix, which is motivated by the numerous potential applications in catalysis, magnetism, optics, I crystallizes in the monoclinic space group P21/c. Its asymmetric unit contains one AgI cation, one [VVO2F2]\u2212 anion and two organic ligands (tr-CH2Ph), which, after inversion across a center of symmetry, form the mol\u00adecular tetra\u00adnuclear cluster {AgI2(VVO2F2)2(tr-CH2Ph)4} \u2005\u00c5; symmetry code (i) \u2212x, \u2212y\u00a0+\u00a01, \u2212z], while the other two link Ag and V centers [the Ag\u22efV distance is 3.8044\u2005(6)\u2005\u00c5]. Thus, the coordination environment of the AgI cation can be described as [AgN3O] with typical Ag\u2014N(triazole) bond lengths [in the range of 2.197\u2005(2) \u2013 2.390\u2005(3)\u2005\u00c5] and a slightly elongated Ag\u2014O bond [2.562\u2005(2)\u2005\u00c5] and 1.8330\u2005(18)\u2005\u00c5], two short V\u2014O [1.632\u2005(2) and 1.660\u2005(2)\u2005\u00c5] and elongated V\u2014N [2.203\u2005(2)\u2005\u00c5] bonds \u2005\u00c5, symmetry code (v) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z] hydrogen-bond contacts. The other triazole group, which provides the heterometallic Ag\u2013V linkage, forms bifurcated C\u2014H\u22efO and C\u2014H\u22efF contacts with vanadium oxofluoride anions of neighboring mol\u00adecular complexes. Additionally, methyl\u00adene \u2013CH2\u2013 fragments show directed C\u2014H\u22efO and C\u2014H\u22efF contacts to the VOF fragments. The phenyl rings are here oriented towards each other in an edge-to-face C\u2014H\u22ef\u03c0 inter\u00adaction mode.Since the organic ligand contains a hydro\u00adphobic benzyl tail, the crystal structure of s Figs.\u00a02. The cenet al., 2004CrystalExplorer17 to 1.385 (blue) a.u. visualizes the set of shortest inter\u00admolecular contacts contacts suggests the dominant role of the hydrogen-bond inter\u00adactions. The strongest ones have a similar nature and they are reflected by pairs of spikes pointing to the lower left of the plot. However, the contribution from the contacts with F-atom acceptors is higher (15.6% for F\u22efH/H\u22efF and 11.6% for O\u22efH/H\u22efO) and they are also essentially shorter, as indicated by different lengths of the spikes (the shortest contacts are F\u22efH = 2.0 and O\u22efH = 2.2\u2005\u00c5). One may suppose that the preferable sites for hydrogen bonding of the vanadium oxofluoride groups are the F atoms. This is consistent with the results of Hirshfeld analysis for the [VOF5]2\u2212 anion 4,4\u2032-bis\u00ad salt with the polarized methyl\u00adene H atoms. There are no mutual \u03c0\u2013\u03c0 inter\u00adactions involving phenyl rings, which are responsible for larger fractions of the C\u22efC contacts in the case of polycyclic species and C\u22efH/H\u22efC (22.2%) contacts, which appear as pairs of nearly identical, very diffuse and short features (N\u22efH = 2.9 and C\u22efH = 2.9\u2005\u00c5). Both of them correspond to edge-to-face stacking or C\u2014H\u22efet al., 2016H-1,2,4-triazol-4-yl) and it revealed five hits for coordination compounds based on this ligand. There are no examples of AgI compounds, only two FeII complexes [FAYQAA ligand.A structure survey was carried out in the Cambridge Structural Database (tr-CH2Ph) was synthesized by refluxing benzyl\u00adamine and di\u00admethyl\u00adformamide azine in the presence of toluene\u00adsulfonic acid monohydrate as a catalyst in DMF (30.0\u2005ml).4-Benzyl-, tr-CH2Ph , V2O5 and 5\u2005mL of water with aqueous HF was added into a Teflon vessel. Then the components were heated at 423\u2005K for 24\u2005h and slowly cooled to room temperature over 50\u2005h, yielding light-yellow prisms of I .Compound 2C6H5 linkage and similarity restraints applied to the closely separated contributions of C12 and C12A, C13 and C13A. H atoms were positioned geometrically and refined as riding, with C\u2014H = 0.93\u2005\u00c5 (CH) and 0.97\u2005\u00c5 (CH2) and with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989022001712/dj2039sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022001712/dj2039Isup2.hklStructure factors: contains datablock(s) I. DOI: 2151864CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The pyran ring is modestly non-planar while the tetra\u00adhydro\u00addiazepine ring adopts a boat conformation. In the crystal, N\u2014H\u22efO hydrogen bonds and slipped \u03c0\u2013\u03c0 stacking inter\u00adactions build a three-dimensional network structure. S,E)-3-diazepin-2-yl\u00adidene]-6-methyl-2H-pyran-2,4(3H)-dione}, C19H16N2O4, is constructed from a benzodiazepine ring system linked to furan and pendant di\u00adhydro\u00adpyran rings, where the benzene and furan rings are oriented at a dihedral angle of 48.7\u2005(2)\u00b0. The pyran ring is modestly non-planar [largest deviation of 0.029\u2005(4)\u2005\u00c5 from the least-squares plane] while the tetra\u00adhydro\u00addiazepine ring adopts a boat conformation. The rotational orientation of the pendant di\u00adhydro\u00adpyran ring is partially determined by an intra\u00admolecular N\u2014HDiazp\u22efODhydp (Diazp = diazepine and Dhydp = di\u00adhydro\u00adpyran) hydrogen bond. In the crystal, layers of mol\u00adecules parallel to the bc plane are formed by N\u2014HDiazp\u22efODhydp hydrogen bonds and slipped \u03c0\u2013\u03c0 stacking inter\u00adactions. The layers are connected by additional slipped \u03c0\u2013\u03c0 stacking inter\u00adactions. A Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (46.8%), H\u22efO/O\u22efH (23.5%) and H\u22efC/C\u22efH (15.8%) inter\u00adactions, indicating that van der Waals inter\u00adactions are the dominant forces in the crystal packing. Computational chemistry indicates that in the crystal the N\u2014H\u22efO hydrogen-bond energy is 57.5\u2005kJ\u2005mol\u22121.The title compound {systematic name: ( In the seven-membered ring, N1 and N2 are displaced from the C1\u2013C6 plane by 0.159\u2005(6) and 0.158\u2005(6)\u2005\u00c5, respectively, in the direction away from C8 = 0.915\u2005(4)\u2005\u00c5, Q(3) = 0.187\u2005(4)\u2005\u00c5, \u03c6(2) = 38.9\u2005(2)\u00b0 and \u03c6(3) = 156.3\u2005(12)\u00b0 [total puckering amplitude Q = 0.933\u2005(4)\u2005\u00c5]. This ring adopts a boat conformation. The mean plane of the O1/C10\u2013C14 ring is inclined to that of the C1\u2013C6 ring by 34.8\u2005(1)\u00b0, while the C1\u2013C6 and O4/C16\u2013C19 rings make a dihedral angle of 48.7\u2005(2)\u00b0. The orientation of the O1/C10\u2013C14 ring is partially determined by an intra\u00admolecular N1\u2014H1\u22efO2 hydrogen bond hydrogen bonds distance = 3.690\u2005(2)\u2005\u00c5, slippage = 1.47\u2005\u00c5] , the white surface indicates contacts with distances equal to the sum of van der Waals radii, and the red and blue colours indicate distances shorter or longer than the van der Waals radii, respectively , while the red regions indicate the negative electrostatic potential (hydrogen-bond acceptors). The shape-index of the HS is a tool to visualize the \u03c0\u2013\u03c0 stacking by the presence of adjacent red and blue triangles. Fig.\u00a04c clearly suggests that there are \u03c0\u2013\u03c0 inter\u00adactions in (I)a, and those delineated into H\u22efH, H\u22efO/O\u22efH, H\u22efC/C\u22efH, C\u22efC, H\u22efN/N\u22efH, C\u22ef O/O\u22efC and O\u22efO contacts have tips at de + di = 2.95\u2005\u00c5. The C\u22efC contacts have an arrow-shaped distribution of points with its tip at de = di = 1.65\u2005\u00c5. The H\u22efN/N\u22efH contacts have tips at de + di = 2.78\u2005\u00c5. Finally, the C\u22efO/O\u22efC and O\u22efO contacts appear with tips at de + di = 3.50\u2005\u00c5 and de = di = 1.73\u2005\u00c5, respectively.In order to visualize the inter\u00admolecular inter\u00adactions in the crystal of (I)rm Fig.\u00a04a, the wH Table\u00a02 contributs Fig.\u00a05d, 15.8%ts Fig.\u00a05e, 7.4% ts Fig.\u00a05f, 2.8% \u22efC Fig.\u00a05g and O\u22ef\u22efO Fig.\u00a05h contacdnorm plotted onto the surface are shown for the H\u22efH, H\u22efO/O\u22efH, H\u22efC/C\u22efH and C\u22efC inter\u00adactions in Fig.\u00a06a\u2013d, respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efO/O\u22efH, and H\u22efC/C\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions play the major role in the crystal packing is the sum of electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies as \u221232.6 (Eele), \u22127.4 (Epol), \u221260.8 (Edis), 57.3 (Erep) and \u221257.5 (Etot).The inter\u00admolecular inter\u00adaction energies were calculated using the CE\u2013B3LYP/6\u201331G energy model available in et al., 2016Hbenzo[b] diazepines substituted at the 2- and 4-positions gave a substantial number of hits with seven deemed closely similar to the title mol\u00adecule C6H3 and R\u2032 = 6- methyl-2H-pyran-2,4-(3H)-dione as well as R = 6-methyl-2H- pyran-2,4-(3H)-dione and R\u2032 = 3-BrC6H4 to 0.957\u2005(2)\u2005\u00c5 . The dihedral angles between the mean planes of the benzo rings and those of the ring-containing substituents on the seven-membered ring vary considerably, likely due to packing considerations as the steric bulk of these groups differ markedly.A search of the Cambridge Structural Database \u00adeth\u00adyl]-4-hy\u00addroxy-6-methyl\u00adpyran-2-one (4\u2005mmol) in ethanol (40\u2005ml) were added 1.5 equivalents of furan-2-carboxaldehyde and four drops of tri\u00adfluoro\u00adacetic acid (TFA). The mixture was refluxed for 3\u2005h. Cooling to room temperature induced the precipitation of a yellow solid, which was filtered off, and then washed with 20\u2005ml of cold ethanol. Crystals suitable for X-ray analysis were obtained by recrystallization of the bulk from ethanol solution to afford colourless crystals (yield: 75%).CELL_NOW was used for the final refinement. Hydrogen atoms attached to carbon were included as riding contributions in idealized positions (C\u2014H = 0.95\u20130.99\u2005\u00c5) with Uiso(H) = 1.2\u20131.5Ueq(C). Those attached to nitro\u00adgen were restrained to a target bond length of 0.91\u2005\u00c5 using the DFIX instruction in SHELXL. The displacement ellipsoids of the O1/C10\u2013C14 ring suggest a possible slight disorder in this group, but it does not appear large enough to model with alternate locations of the atoms.Crystal, data collection and refinement details are presented in Table\u00a0310.1107/S2056989021007441/wm5612sup1.cifCrystal structure: contains datablock(s) I, global. DOI: Click here for additional data file.10.1107/S2056989021007441/wm5612Isup3.cdxSupporting information file. DOI: 10.1107/S2056989021007441/wm5612Isup4.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021007441/wm5612Isup4.cmlSupporting information file. DOI: 2097593CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-020-68370-y, published online 21 August 2020Correction to: This Article contains a typographical error in the Results section under subheading \u2018Mechanical properties\u2019 where,\u201cThe compressive strength of the scaffolds calculated as the maximum compression force divided by the cross-sectional area of tested specimen, was 87.4\u2009\u00b1\u200922.1 and 6.5\u2009\u00b1\u20090.8\u00a0MPa for ATE-G3 and Repros (BCP) scaffolds, respectively.\u201dshould read:\u201cThe compressive strength of the scaffolds calculated as the maximum compression force divided by the cross-sectional area of tested specimen, was 8.7\u2009\u00b1\u20092.2 and 6.5\u2009\u00b1\u20090.8\u00a0MPa for ATE-G3 and Repros (BCP) scaffolds, respectively.\u201d"} +{"text": "The daily life of Chronic obstructive pulmonary disease (COPD) patients is characterised not only by chronic respiratory symptoms but also by exercise intolerance due to their breathlessness. Proper diagnosis and management of this disease consequently includes evaluation of exercise tolerance , frequenAn observational study was performed with inclusion of seventy one men with moderate COPD (FEV1 54.6\u2009\u00b1\u20097.1%); age 63.8\u2009\u00b1\u20093.1 yrs; weight, 71.2\u2009\u00b1\u20098.3\u2009kg; height, 169.0\u2009\u00b1\u20098.1\u2009cm constituted the COPD group (COPDG), and 150 healthy subjects, age 64.2\u2009\u00b1\u20095.8 yrs; weight, 76.2\u2009\u00b1\u200911.3\u2009kg; height, 169.8\u2009\u00b1\u20097.5\u2009cm, were included as the healthy group \u2013 HG. The physical parameters assessed were strength, aerobic endurance, flexibility and agility/balance, by the Fullerton\u2019s functional fitness tests. The health status was evaluated through the Medical Outcomes 36-item Short Form Health Survey (SF-36) questionnaire. The study was approved by the Ethics Committee of the Garcia de Orta Hospital and all participants gave their informed consent.p\u2009<\u2009.05) between COPDG and HG groups for the following variables all expressed in mean\u2009\u00b1\u2009SD: body mass index, 25.9\u2009\u00b1\u20093 vs 27.7\u2009\u00b1\u20094.1\u2009kg.m2; 30-second chair stand 14.1\u2009\u00b1\u20091.7 vs. 18.2\u2009\u00b1\u20091.9 times; arm curl 15.7\u2009\u00b1\u20092.8 vs. 18.8\u2009\u00b1\u20094.9 times; 6-minute walk 498.8\u2009\u00b1\u200958.3 vs. 589.7\u2009\u00b1\u200988.6 m; 8-foot up-and-go 4.7\u2009\u00b1\u20090.8 vs. 5.1\u2009\u00b1\u20091\u2009sec; chair sit-and-reach 0.81\u2009\u00b1\u20099.9 vs. \u22127.1\u2009\u00b1\u200910.6\u2009cm respectively and no differences were observed for the back scratch test . In health status DPOCG presented a significant decrease (p\u2009<\u2009.05) on perception of all domains of SF-36, except on body pain.The values of the functional fitness test were significantly different (In this study COPD patients have lower levels of functional capacity compared to healthy subjects. However, they were able to perform short tasks with higher speed. This trend was also evident in other study where COPD patients performed short term activities faster than healthy persons . Limitat"} +{"text": "In Xu et al.,p value\u00a0<\u00a00.05 vs. NC group; NC: negative control). F, Overexpression of lncRNA\u2010CASC7 increased the expression of IL\u201011 in H9C2 cellsFIGURE 9: lncRNA\u2010CASC7 suppressed the expression of miR\u201030c in H9C2 cells (*Supplementary MaterialClick here for additional data file."} +{"text": "A series of platinum(II) complexes with differently substituted iminomethyl groups were synthesized, and their photophysical properties were examined in solution, in the crystalline, and in the PMMA film\u2010dispersed state, respectively (PMMA=poly(methyl methacrylate)). These complexes showed structure\u2010dependent emission spectra, in which the color of the luminescence in the crystalline state varied over a range of about 40\u2005nm depending on the specific bowl\u2010shaped molecular structure. The chiral complexes with \u2010 and \u2010configurations were found to have structure\u2010dependent chiroptical properties both in solution and the PMMA film\u2010dispersed state such that the intensity of circular dichroism (CD) and circularly polarized luminescence (CPL) were enhanced with bulky cyclic substituents at the nitrogen atoms. A theoretical study using density functional theory (DFT) and time\u2010dependent (TD)\u2010DFT calculations revealed that the enhancement of chiroptical responses is due to the amplification of the magnetic dipole moment caused by the distortion of the square planar geometry.The relationship between the coordination geometry and photophysical properties of Chiral platium(II)\u00a0complexes exhibit structure\u2010dependent chiroptical properties in dilute solution and in the poly(methyl methacrylate) film\u2010dispersed state. DFT and TD\u2010DFT calculations of the structures and electronic configurations revealed that the enhancement of the chiroptical responses is due to the distortion of the square planar geometry. Complexes \u2010 and \u20101\u2009a\u2013e were successfully characterized by 1H and 13C nuclear magnetic resonance (NMR) spectroscopy , infrared (IR) spectroscopy, and high\u2010resolution mass spectrometry (HRMS). The same treatment with the racemic ligand yielded a mixture of rac\u2010/\u20101\u2009a and meso\u2010\u20101\u2009a (ca. 1\u2009:\u20091). Recrystallization from CH2Cl2/n\u2010hexane gave enriched \u20101\u2009a in 46\u2009% diastereomeric excess. Repeated crystallization from CH2Cl2/n\u2010hexane yielded pure crystals of \u20101\u2009a. From the mother liquor of the first recrystallization, enriched /\u20101\u2009a in 6\u2009% diastereomeric excess was obtained. Crystallization of the enriched /\u20101\u2009a from CH2Cl2/n\u2010hexane several times afforded pure crystals of /\u20101\u2009a.A series of chiral 1\u2009a\u2013e were obtained by recrystallization from CH2Cl2/n\u2010hexane and employed in single\u2010crystal X\u2010ray diffraction (XRD) analysis. Details of the crystal data and the structure refinement are presented in Table\u2005S1 (Supporting Information), including analysis of the intermolecular interactions . ORTEPS,S)\u20101\u2009a, /\u20101\u2009a, \u20101\u2009a, \u20101\u2009b\u2013d, and \u20101\u2009e are shown in Figures\u2005\u03d5) and bowl angles \u03b8 between the mean planes of the naphthalene rings are provided to express the degree of distortion of the coordination planes.Single crystals of platinum complexes S,S)\u20101\u2009a, /\u20101\u2009a, and \u20101\u2009a are shown in Figures\u2005S,S)\u20101\u2009a has a bowl configuration (\u03b8=49.6\u00b0), and the two methyl groups have a syn\u2010orientation \u20101\u2009a in the spiro\u2010chelate rings has a positive value of 7.94\u00b0. Hence, \u20101\u2009a has a configuration in the crystal. These structural data correspond well with the previously reported palladium(II) complex.R,R)/\u20101\u2009a have an envelope form with a \u03d5 angle of \u22122.64\u00b0 . Therefore, \u20101\u2009a in /\u20101\u2009a has a configuration in the crystal, whilst \u20101\u2009a in the enantiopure crystal showed the opposite \u039b configuration. \u20101\u2009a adopts a step configuration with C2 symmetry, and the two methyl groups in \u20101\u2009a take an anti\u2010orientation .The molecular structures of \u20101\u2009b\u2013d and \u20101\u2009e are shown in Figures\u2005S,S)\u20101\u2009b adopts the bowl configuration (\u03b8=19.2\u00b0) with anti\u2010orientation between the phenyl groups \u20101\u2009c are in anti\u2010orientation, and the structure can be described as a distorted bowl\u2010configuration with a low bowl angle (\u03b8=3.56\u00b0) \u20101\u2009d has a bowl\u2010configuration with \u03b8=48.2\u00b0 as the bowl angle and a \u039b configuration with a positive value of \u03d5=1.30\u00b0 \u20101\u2009e adopts the step configuration (\u03b8=5.14\u00b0) with an anti\u2010orientation between the methyl groups \u2010 and \u20101\u2009a\u2013e were recorded in solution and in the PMMA (PMMA=poly(methyl methacrylate) film\u2010dispersed state at room temperature \u20101\u2009a\u2013d exhibited a strong negative Cotton effect at approximately 420\u2005nm, which can be assigned to a mixture of intra\u2010ligand charge transfer (1ILCT) and metal\u2010to\u2010ligand charge transfer (1MLCT) from the UV/Vis spectra showing a mixture of 1ILCT and 1MLCT bands around 370\u2013500\u2005nm observed under the same measurement conditions \u20101\u2009e showed a comparably weak negative Cotton effect in the same region, although it has almost the same UV/Vis spectrum as \u20101\u2009a\u2013d. The gabs (=\u0394\u03f5/\u03f5) values around the absorption maxima in the low energy region are \u22120.0026 (422\u2005nm) for \u20101\u2009a, \u22120.0023 (422\u2005nm) for \u20101\u2009b, \u22120.0030 (417\u2005nm) for \u20101\u2009c, \u22120.0025 (418\u2005nm) for \u20101\u2009d, and \u22120.00088 (410\u2005nm) for \u20101\u2009e, respectively. These results indicate that the bulkiness of the substituents at the nitrogen atoms exert a strong influence on the chiral properties of 1 even in the diluted solution state.Circular dichroism (CD) spectra of \u20101\u2009d (\u03bbmax=589\u2005nm) with the bulkiest 1\u2010naphthyl substituent at the nitrogen atoms compared to those of \u20101\u2009a\u2013c \u20101\u2009a, 601\u2005nm for \u20101\u2009b, and 602\u2005nm for \u20101\u2009c, respectively). In contrast, a bathochromic shift was observed for \u20101\u2009e (\u03bbmax=608\u2005nm) with the less bulky alkyl substituents at the nitrogen atoms. These results indicate that the bulkiness of the substituents at the nitrogen atoms affects the d\u2010\u03c0 conjugation of the coordination platform in dilute solution.All the complexes exhibited orange to red luminescence under UV excitation at room temperature in solution, crystalline, and PMMA film\u2010dispersed state, respectively . The photophysical data of complexes S,S)\u20101\u2009a\u2013e was more remarkable in the crystalline state. Figure\u2005S,S)\u20101\u2009a\u2013e in the crystalline state, where the emission peak maxima clearly changes depending on the substituent at the nitrogen atoms \u20101\u2009a, 623\u2005nm for \u20101\u2009b, 605, 628\u2005nm for \u20101\u2009c, 605\u2005nm for \u20101\u2009d, and 632\u2005nm for \u20101\u2009e).The change in the emission color of /\u20101\u2009a and \u20101\u2009a showed bathochromic emission patterns that were different from that of enantiopure crystals \u20101\u2009a . XRD analysis showed that the maximum emission wavelength (\u03bbmax) in the crystalline state of complexes 1 was negatively correlated with the bowl angle . Therefore, we assume that bowl\u2010shaped structures exert a strong influence on the d\u2010\u03c0 conjugation of 1, resulting in a change in the emission color in the crystalline state.It is noteworthy that the crystals of (I) and total luminescence (I) spectra measured for \u2010 and \u20101\u2009a\u2013e in the PMMA film\u2010dispersed state at 298\u2005K are shown in Figures\u2005R,R)\u2010 and \u20101\u2009a\u2013d exhibit mirror\u2010image positive and negative signals around 570\u2013590\u2005nm, which are corresponding to the emission maxima (\u03bbmax) observed in their total emission spectra taken under the same measurement conditions for \u20101\u2009a, \u22120.0011 (575\u2005nm) for \u20101\u2009b, \u22120.0036 (590\u2005nm) for \u20101\u2009c, and \u22120.0018 (575\u2005nm) for \u20101\u2009d, respectively. It should be noted that \u2010 and \u20101\u2009e did not exhibit detectable CPL signals , which indicates that circularly polarized luminescence was induced by introduction of bulky substituents at the nitrogen atoms.The circularly polarized luminescence \u20101\u2009a\u2013e in an isolated system were estimated using DFT calculations on the basis of the X\u2010ray structures. The optimized structures of \u20101\u2009a\u2013e are shown in Figure\u2005\u03d5) are 0.90\u00b0 for 1\u2009a, 0.78\u00b0 for 1\u2009b, 0.29\u00b0 for 1\u2009c, 1.39\u00b0 for 1\u2009d, and 0.20\u00b0 for 1\u2009e, indicating that the substituents on the nitrogen atoms distort the coordination geometry which induces \u039b chirality. In addition, \u20101\u2009a\u2013d have a bowl\u2010configuration with similar bowl angles \u03b8, while \u20101\u2009e has a step\u2010configuration with a small \u03b8 value. These structural differences are expected to affect the chiroptical properties in solution and in the PMMA film\u2010dispersed state.In order to clarify the structure\u2010dependence of the photophysical properties of the series of platinum complexes S,S)\u20101\u2009a\u2013e and their eigenvalues were estimated by using DFT calculations on the basis of the optimized structure in the ground state. The HOMOs of all complexes are principally Pt(dzx)\u2013ligand (\u03c0) hybrids, whereas the LUMOs are in the ligand (\u03c0*) states is the HOMO\u2010to\u2010LUMO transition, which implies that the present emission is principally attributable to a mixture of MLCT/ILCT. The S0\u2013T1 transition energies were calculated to be 2.17\u2005eV (570\u2005nm) for 1\u2009a, 2.18\u2005eV (568\u2005nm) for 1\u2009b, 2.19\u2005eV (567\u2005nm) for 1\u2009c, 2.21\u2005eV (561\u2005nm) for 1\u2009d, and 2.14\u2005eV (579\u2005nm) for 1\u2009e, which is consistent with the hypsochromic and bathochromic shift of the emission peak maxima for 1\u2009d and 1\u2009e in CH2Cl2 calculations (Table\u2005S2). The major consequence of the electronic configuration of the S 2.17\u2005eV 70\u2005nm forS,S)\u20101\u2009a\u2013e using TD\u2010DFT calculations . The validity of the calculation was confirmed by comparison between theoretical and experimental CD spectra of \u20101\u2009a\u2013e /(|\u03bce|2+|\u03bcm|2), where |\u03bce|, |\u03bcm|, and \u03b8e,m are the electric transition dipole moments, magnetic transition dipole moments, and the angles between two vectors \u03bce and \u03bcm, respectively. , corresponding chiral ligands (0.58\u2005mmol), and K2CO3 (1.5\u2005mmol) in toluene/DMSO (20/5\u2005mL) was warmed at 80\u2009\u00b0C for 16\u2005h. The red reaction mixture was added to water (30\u2005mL) and extracted with EtOAc (20\u2005mL\u00d73). The organic layers were dried over Na2SO4 and evaporated in vacuo. The residue was washed with cold methanol to give the complexes. Suitable crystals for the X\u2010ray analysis were prepared by recrystallization from CH2Cl2/n\u2010hexane.transN\u2010Bis[1\u2010[(1\u2010phenylethyl)imino]methyl\u20102\u2010naphthalenolato\u2010,OR]\u2010platinum(II) \u2010 and \u20101\u2009a): Orange crystals (38\u2009%); m.p. 230\u2009\u00b0C (decomp); IR (KBr): \u03bd=1616\u2005cm\u22121 (N=C); 1H NMR \u03b4=8.64 , 7.67 , 7.64 , 7.59 , 7.44 , 7.39\u20137.32 , 7.19 , 7.10 , 6.36 , 1.93\u2005ppm ; 13C\u2005NMR \u03b4=163.03, 152.69, 142.30, 134.00, 133.82, 128.80, 128.73, 128.12, 127.71, 127.39, 127.33, 123.23, 122.48, 119.16, 111.65, 58.53, 21.76\u2005ppm; HRMS (ESI+): m/z [M+H]+ calcd. for C38H33N2O2195Pt: 742.2092, found 742.2094; Anal. Calcd for C38H32N2O2Pt: C, 61.37; H, 4.34; N, 3.77. Found: C, 61.21; H, 4.33; N, 3.65. \u20101\u2009a: [\u03b1]D25=\u2212166 .transN\u2010Bis[1\u2010[(1\u2010phenylpropyl)imino]methyl\u20102\u2010naphthalenolato\u2010,OR]\u2010platinum(II) \u2010 and \u20101\u2009b): Orange crystals (36\u2009%); m.p. 254\u2009\u00b0C (decomp); IR (KBr): \u03bd=1614\u2005cm\u22121 (N=C); 1H NMR \u03b4=8.66 , 7.69 , 7.66 , 7.57 , 7.45\u20137.32 , 7.20 , 7.14 , 6.10 , 2.53\u20132.23 , 1.18\u2005ppm ; 13C\u2005NMR \u03b4=162.80, 152.47, 140.52, 133.94, 133.91, 128.88, 128.75, 127.79, 127.37, 127.33, 123.20, 122.44, 119.17, 111.62, 64.51, 28.64, 11,70\u2005ppm; HRMS (APCI+): m/z [M+H]+ calcd. for C40H37N2O2195Pt: 772.2501, found 772.2513; Anal. calcd for C40H36N2O2Pt: C, 62.25; H, 4.70; N, 3.63. Found: C, 62.25; H, 4.67; N, 3.68. \u20101\u2009b: [\u03b1]D25=\u2212231 .transN\u2010Bis[1\u2010[(1\u2010cyclohexylethyl)imino]methyl\u20102\u2010naphthalenolato\u2010,OR]platinum(II) \u2010 and \u20101\u2009c): Orange crystals (23\u2009%); m.p. 278\u2009\u00b0C (decomp); IR (KBr): \u03bd=1616\u2005cm\u22121 (N=C); 1H NMR \u03b4=8.63 , 7.81 , 7.70 , 7.51 , 7.25 , 7.10 , 4.72 , 2.08\u20131.65 , 1.50 , 1.35\u20131.15\u2005ppm ; 13C\u2005NMR \u03b4=163.01, 150.76, 133.98, 133.61, 128.83, 127.39, 127.30, 123.20, 122.39, 119.30, 111.74, 61.77, 43.98, 30.43, 28.64, 26.51, 26.29, 26.23, 18.72 ppm; HRMS (APCI+): m/z [M+H]+ calcd. for C38H45N2O2195Pt: 756.3127, found 756.3149; Anal. calcd for C38H44N2O2Pt\u2009\u22c5\u20091/2CH2Cl2: C, 57.92; H, 5.68; N, 3.51. Found: C, 57.83; H, 5.19; N, 3.39. \u20101\u2009c: [\u03b1]D25=\u2212153 transN\u2010Bis[1\u2010[ethyl)imino]methyl\u20102\u2010naphthalenolato\u2010,OR]platinum(II) \u2010 and \u20101\u2009d): Orange crystals (48\u2009%); m.p. >300\u2009\u00b0C; IR (KBr): \u03bd=1615\u2005cm\u22121 (N=C); 1H NMR \u03b4=8.75\u20138.72 , 8.65 , 7.96 , 7.92\u20137.87 , 7.67 , 7.60 , 7.57 , 7.52\u20137.46 , 7.22 , 7.13\u20137.09 , 6.99 , 2.14\u2005ppm ; 13C\u2005NMR \u03b4=163.19, 151.27, 137.26, 134.27, 133.93, 133.76, 131.76, 129.23, 128.61, 128.53, 127.28, 127.22, 127.17, 126.08, 125.55, 125.12, 124.87, 123.09, 122.37, 119.01, 111.81, 55.67, 22.64\u2005ppm; HRMS (APCI+): m/z [M+H]+ calcd. for C46H37N2O2195Pt: 844.2501, found 844.2479; Anal. calcd for C46H36N2O2Pt\u2009\u22c5\u2009CH2Cl2: C, 60.78; H, 4.12; N, 3.02. Found: C, 60.79; H, 4.33; N, 2.83. \u20101\u2009d: [\u03b1]D25=\u2212243 .transN\u2010Bis[1\u2010[(1\u2010methyllpropyl)imino]methyl\u20102\u2010naphthalenolato\u2010,OR]platinum(II) \u2010 and \u20101\u2009e): Orange crystals (34\u2009%); m.p. 264\u2009\u00b0C (decomp); IR (KBr): \u03bd=1608\u2005cm\u22121 (N=C); 1H NMR \u03b4=8.72 , 7.83 , 7.70\u20137.68 , 7.50 , 7.26\u20137.23 , 7.10 , 4.88\u20134.81 , 2.15\u20132.06 , 1.85\u20131.76 , 1.54 , 1.09 ppm ; 13C\u2005NMR \u03b4=163.02, 150.11, 133.95, 133.67, 128.83, 127.41, 127.29, 123.18, 122.41, 119.24, 111.71, 58.46, 31.10, 21.46, 11.02 ppm; HRMS (APCI+): m/z [M+H]+ calcd. for C30H33N2O2195Pt: 648.2187, found 648.2185. \u20101\u2009e: [\u03b1]D25=\u221227 .RPreparation of /(S)\u2010 and imino]\u2010methyl\u20102\u2010naphthalenolato\u2010N),OR]platinum(II) /(S)\u20101\u2009a and \u20101\u2009a: A solution of [PtCl2(CH3CN)2] , 2\u2009a , and K2CO3 in toluene/DMSO (20/5\u2005mL) was warmed at 80\u2009\u00b0C for 16\u2005h. The dark red reaction mixture was added to water (30\u2005mL) and extracted with EtOAc (20\u2005mL\u00d73). After evaporation of the solvent, the residue was washed with cold methanol to give a mixture of \u20101\u2009a and /\u20101\u2009a (51\u2009:\u200949 ratio) in 50\u2009% (213\u2005mg) yield. From the first recrystallization of CH2Cl2/n\u2010hexane, the filtrate gave a \u20101\u2009a enriched powder (46\u2009% de). Pure \u20101\u2009a was obtained by recrystallization of CH2Cl2/n\u2010hexane after several times, crystals of which were suitable for X\u2010ray analysis. After evaporation of the first mother liquor, a /\u20101\u2009a enriched orange powder (6\u2009% de) was obtained. Pure /\u20101\u2009a was obtained by recrystallization of CH2Cl2/n\u2010hexane after several times, crystals of which were suitable for X\u2010ray analysis. Pure crystals of /\u20101\u2009a were prepared by crystallization of an 1\u2009:\u20091 mixture of \u20101\u2009a and \u20101\u2009a in CH2Cl2/n\u2010hexane.R\u20101\u2009a: m.p. 265\u2009\u00b0C (decomp); IR (KBr): 1606\u2005cm\u22121 (N=C). 1H NMR : \u03b4=8.64 , 7.67 , 7.64 , 7.61 , 7.45 , 7.39\u20137.34 , 7.19 , 7.10 , 6.36 , 1.90\u2005ppm . 13C\u2005NMR : \u03b4=163.03, 152.68, 142.26, 134.00, 133.83, 128.83, 128.74, 128.17, 127.73, 127.39, 127.33, 123.21, 122.48, 119.16, 111.65, 58.69, 21.76\u2005ppm. HRMS (ESI+): m/z [M+H]+ calcd. for C38H33N2O2195Pt: 745.2184, found 745.2147.R,R)/\u20101\u2009a The NMR and IR spectra corresponded well with the data of \u2010 and \u20101\u2009a. HRMS (ESI+): m/z [M+H]+ calcd. for C38H33N2O2195Pt: 745.2184, found 745.2156.X\u2010Ray Crystallography: Crystals suitable for XRD studies were analyzed using a Rigaku RAXIS\u2010RAPID imaging plate diffractometer using Mo\u2010K\u03b1 radiation and a Rigaku XtaLAB mini2 benchtop X\u2010ray crystallography system equipped with a Mo rotating\u2010anode X\u2010ray generator with monochromated Mo\u2010K\u03b1 radiation (\u03bb=0.71073\u2005\u00c5). The molecular structures and packings in crystals \u20101\u2009a\u2013d, \u20101\u2009e, /\u20101\u2009a, and \u20101\u2009a were solved by direct methods and refined using the full\u2010matrix least\u2010squares method. In subsequent refinements, the function \u03a3\u03c9(Fo2\u2212Fc2)2 was minimized, where Fo and Fc are the observed and calculated structure factor amplitudes, respectively. The positions of non\u2010hydrogen atoms were found from difference Fourier electron density maps and refined anisotropically. All calculations were performed using the Crystal Structure crystallographic or CrysAlisPro program software package, and illustrations were drawn by using ORTEP.S,S)\u20101a), 1918308 /\u20101a), 1918309 \u20101a), 1918310 \u20101b), 1918311 \u20101c), 1918312 \u20101d), 2119929 \u20101e)1918306 ) and singlet\u2013triplet (E(Tn)) transition energies were estimated by time\u2010dependent (TD) DFT calculation .The authors declare no conflict of interest.1As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efS hydrogen bonds, forming a two-dimensional supra\u00admolecular architecture. 3H5OS2)2(C6H16N2)] or [Hg(C3H5OS2)2(tmeda)] , has a distorted tetra\u00adhedral geometry. The HgII ion is coordinated to two N atoms of the N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adethylenedi\u00adamine ligand and two S atoms from two ethylxanthate xanthate ligands. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efS hydrogen bonds, forming a two-dimensional supra\u00admolecular architecture in the ab plane. The most important contributions for the crystal packing are from H\u22efH (59.3%), S\u22efH (27.4%) and O\u22efH (7.5%) inter\u00adactions.The title four-coordinate mononuclear complex, [Hg(C The bond lengths and angles of the HgN2S2 coordination units correspond to those in the structures of mixed-ligand HgII coordination compounds (see Database survey). The C1\u2014O1 and C2\u2014O1 bond lengths are 1.355\u2005(11) to 1.460\u2005(12)\u2005\u00c5, respectively, although all of the C\u2014O bonds show single-bond character. In the {S2C} section of the xanthate ligands, the C1\u2014S1 distance is 1.727\u2005(9)\u2005\u00c5, which is typical of a single bond, whereas the C1=S2 distance of 1.633\u2005(10)\u2005\u00c5 is typical of a carbon-to-sulfur double bond. The C\u2014N and C\u2014C bond lengths in the N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adethylenedi\u00adamine ligand are normal 2(tmeda)].In the crystal, there is a weak inter\u00admolecular hydrogen bonding Table\u00a02 between et al., 201614H26O2S4)]n 2(C6H16N2)] (C9H13NS)2](PF6)2 (C6H16N2)] ] 2 in ethanol (10\u2005mL) under stirring. The formed precipitate was filtered off, washed with water and air-dried. The precipitate was suspended in hot ethanol (10\u2005mL) and tetra\u00admethyl\u00adethylenedi\u00adamine was added under stirring. The colour changed to dark brown. The precipitate was filtered off and dried and then recrystallized from ethanol. Brown rods were formed.Potassium ethylxanthate in hot ethanol (10\u2005mL) was added to a hot solution of Hg(CHUiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for all othersCrystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021010549/ey2008sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021010549/ey2008Isup2.hklStructure factors: contains datablock(s) I. DOI: 2115100CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The sheets are associated through additional C\u2014H\u22ef\u03c0(ring) inter\u00adactions.The conformation of the title compound is partially determined by a strong, intra\u00admolecular O\u2014H\u22efO hydrogen bond. In the crystal, C\u2014H\u22efO hydrogen bonds link the mol\u00adecules, forming chains along the 13H11NO2, is partially determined by a strong, intra\u00admolecular O\u2014H\u22efO hydrogen bond. The crystal packing consists of strongly corrugated layers parallel to the ac plane and associated through C\u2014H\u22ef\u03c0(ring) inter\u00adactions. A Hirshfeld surface analysis of the crystal structure indicates that the most significant contributions to the crystal packing are from H\u22efH (44.1%), C\u22efH/H\u22efC (29.4%) and O\u22efH/H\u22efO (17.3%) contacts.The conformation of the title compound, C They show anti\u00adfungal is 1.94\u2005(12) \u00b0. The torsion angles C2\u2014C1\u2014C7\u2014N1, C1\u2014C7\u2014N1\u2014C8, C1\u2014C7\u2014N1\u2014O2, C7\u2014N1\u2014C8\u2014C9 and O2\u2014N1\u2014C8\u2013C-9 are \u221230.2\u2005(3), \u2212179.7\u2005(2), \u22120.4\u2005(3), 27.3\u2005(3) and \u2212152.0\u2005(2)\u00b0, respectively. The conformation of the title compound is partially determined by a strong, intra\u00admolecular O1\u2014H1\u22efO2 hydrogen bond -N-[methanimine]-N-oxido -1,2-bis\u00ad(3-bromo\u00adphen\u00adyl)diazene 1-oxide -N-benzyl\u00adidene-1-phenyl\u00admethanamine oxide hydrogen peroxide solvate -N-(2-chloro\u00adbenzyl\u00adidene)aniline N-oxide \u00b0] and the phenyl and benzene rings are trans-oriented around the C=N bond. The phenyl and benzene rings make a dihedral angle of 56.99\u2005(2)\u00b0.In the crystal of DEPVOM, (101) layers are generated by C\u2014H\u22efO hydrogen bonds coupled with C\u2014H\u22ef\u03c0(ring) and offset \u03c0\u2013\u03c0 stacking inter\u00adactions. In the crystal of SIYHAK01, C\u2014H\u22efO and C\u2014H\u22efBr hydrogen bonds together with offset \u03c0\u2013\u03c0 inter\u00adactions stack the mol\u00adecules along the Z)-(2-Hy\u00addroxy\u00adphen\u00adyl)methyl\u00adidene]benzenimine N-oxide (nitrone) was prepared according to the reported procedures = 1.5Ueq(O). The C-bound H atoms were positioned geometrically, with C\u2014H = 0.95\u2005\u00c5, and constrained to ride on their parent atoms, withUiso(H) = 1.2Ueq(C). Attempts to determine the absolute structure did not produce a definitive result, viz.: Flack x = 0.2\u2005(3) by classical fit to all intensities 0.30\u2005(14) from 611 selected quotients (Parsons\u2019 method). A round of TWIN/BASF refinement gave BASF = 0.2\u2005(4) with no improvement in the model.Crystal and refinement details are presented in Table\u00a0410.1107/S2056989021004813/ey2006sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989021004813/ey2006Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021004813/ey2006Isup3.cmlSupporting information file. DOI: 2082055CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In addition, the mononuclear Dy and Tb complexes [{(thf)2Li(NtBu)2S(tBuN)2LnCl2(thf)2] show slow magnetic relaxation under applied dc fields.Lanthanide ions are particularly well\u2010suited for the design of single\u2010molecule magnets owing to their large unquenched orbital angular momentum and strong spin\u2010orbit coupling that gives rise to high magnetic anisotropy. Such nanoscopic bar magnets can potentially revolutionize high\u2010density information storage and processing technologies, if blocking temperatures can be increased substantially. Exploring non\u2010classical ligand scaffolds with the aim to boost the barriers to spin\u2010relaxation are prerequisite. Here, the synthesis, crystallographic and magnetic characterization of a series of each isomorphous mono\u2010 and dinuclear lanthanide complexes comprising tetraimido sulfate ligands are presented. The dinuclear Dy complex [{(thf) The first series of isomorphous lanthanide Gd to Er complexes containing the tetraimido sulfate ligand renders these compounds compelling to be investigated for their single\u2010molecule magnet behaviour. The displayed Dy dimer shows slow magnetic relaxation at zero field. The monomeric Tb and Dy complexes show field\u2010induced single\u2010molecule magnet behaviour. The ligand holds great potential for higher blocking systems, that is, removal of chloride ions, lithium transmetalation and promotion of the magnetic communication between the metal ions. S\u2010alkyltriimidosulfonates [RS(NR)3]\u2212 and in the triimidosulfonic acid MeS(NtBu)2NHtBu the sum of all three S\u2212N bond lengths is constant at 4.70(2)\u2005\u00c5.[3]2\u2212 it was later also found in the tetraimido sulfates [S(NR)4]2\u2212.tBu)]\u2212 ligand in [Co{(NtBu)3SMe}2], ,tBu)4]2\u2212 anion.[Single\u2010molecule magnets (SMMs)4Li2(NtBu)4S] with the appropriate lanthanide(III) chloride in thf yields after extraction and crystallization from toluene the binuclear lanthanide(III) compounds [{(thf)2Li(NtBu)2S(tBuN)2LnCl2}2\u2009\u22c5\u2009ClLi(thf)2] 1\u2009a\u2013e with a: Ln=Gd, b: Tb, c: Dy, d: Ho, e: Er 2Li(NtBu)2S(tBuN)2LnCl2(thf)2] 2\u2009a\u2013e in good yields and purity. Instead, 2\u2009c was also isolated upon layering the reaction mixture with n\u2010pentane. However, since the crystallization parameters for this reaction are very tedious to control and the final product is insoluble in thf the derivatives rather precipitate than crystallize. Therefore, we concentrated on the optimization of the synthetic route to give the \u03bc\u2010LiCl\u2010bridged dimers 1\u2009a\u2013e. They crystallize in the monoclinic space group C2/c with half a molecule and half a toluene molecule in the asymmetric unit 42\u2212, the S\u2212N bond distances for all metal complexes are almost identical. Nevertheless, it should be stated that the av. S\u2212N1/N2 distances from the lithium coordinating nitrogen atoms of 1.567\u2005\u00c5 are significantly shorter than the S\u2212N3/N4 distances of 1.626\u2005\u00c5 to the lanthanide\u2010coordinated nitrogen atoms. That mirrors the two opposite coordination sites of the tetrahedral ligand which are unsymmetrically coordinated by a lithium ion at the site with shorter S\u2212N bonds and the lanthanide(III) ion at the opposite site. The triply positive charged lanthanides obviously are in much stronger demand of the negatively charged nitrogen atoms, for their part taking advantage of the electropositive central sulfur atom. A stronger demand of the negatively charged nitrogen atoms causes a longer distance between the nitrogen and the positively charged sulfur. The reduced lanthanide radii are also displayed in the reduction of the Ln1\u22c5\u22c5\u22c5Ln1A distances that ranges from 3.835\u2005\u00c5 in 1\u2009a to 3.752\u2005\u00c5 in 1\u2009e. Interestingly, the reduced radii have the opposite effect on the N1\u2212Ln1\u2212N2 angles increasing from 60.91\u00b0 in 1\u2009a to 62.24\u00b0 in 1\u2009e. Even though all Cl\u2212Ln\u2212Cl angles for one specific metal are different, they remain almost identical when proceeding from one lanthanide cation to the next. The same is valid for the N\u2212S1\u2212N angles. A comparable coordination is found in a series of rare earth metal complexes with \u03b2\u2010diketiminato ligands. Substitution at the nitrogen position with 2,6\u2010dimethylphenyl,1\u2009c with 2.726\u2005\u00c5. The same is valid for the Dy\u2212N bond lengths of 2.313\u2005\u00c5 and 2.270\u2005\u00c5, respectively.We found these dimeric complexes, dissolved in small amounts of thf to recrystallize within hours at room temperature, forming the mononuclear lanthanide(III) compounds 3\u2009b could be isolated from toluene . All Tb\u2212Cl distances are similar, although the terbium atoms in 3\u2009b are fourfold chlorine coordinated and those in 1\u2009b and 2\u2009b only two\u2010fold. With the rising nuclearity in 3\u2009b the Tb\u039bTb distance increases to 4.307\u2005\u00c5 compared to 3.831\u2005\u00c5 in 1\u2009b. The acute N1\u2212Tb\u2212N2 bite angles of 61.22\u00b0 (1\u2009b), 60.67\u00b0 (2\u2009b), and 61.10\u00b0 (3\u2009b) differ only marginally. The same is valid for the N1\u2212S1\u2212N2 ligand angles at the Tb side of 91.47\u00b0 (1\u2009b), 92.77\u00b0 (2\u2009b) and 92.60\u00b0 (3\u2009b).Tetranuclear 1\u2009a\u2013e and 2\u2009b\u2013c in the temperature range 2\u2013300\u2005K at 0.1, 0.5 and 1 T fields are in good agreement with the expected values for two non\u2010interacting triply positive lanthanide atoms. As the temperature is decreased to 15\u2005K, \u03c7MT undergoes a gradual decline and decreases below that temperature rapidly to reach at 2\u2005K a minimum value of 10.24, 8.99, 9.57, 9.14, and 6.46\u2005cm3K/mol for 1\u2009a\u2013e, respectively. Such downturn in \u03c7MT can be generally assigned to the Zeeman effect and/or weak intermolecular interactions. Notably, the decline in \u03c7MT is much more pronounced for 1\u2009e and can potentially be ascribed to quite anisotropic trivalent erbium atoms in this ligand environment compared to the lighter lanthanide analogs. By contrast, \u03c7MT only decreases more noticeably below 10\u2005K for 1\u2009a hinting at the presence of very weak antiferromagnetic coupling. Due to the half\u2010filled f\u2010electron valence shell (4f7), trivalent gadolinium ions offer the possibility to determine the magnetic exchange interaction precisely. Fitting the dc data of 1\u2009a to a spin\u2010only Hamiltonian \u0124=\u22122J\u015cGd(1)+\u22122J\u015cGd(2) where J is the intramolecular coupling constant and \u015c the spin operator for each paramagnetic center yielded a coupling constant of J=\u22120.045(1)\u2005cm\u22121 which is indicative of weak antiferromagnetic interaction between the GdIII ions. The magnitude of the coupling between the lanthanide ions via superexchange is typically very weak as is the case for the parent compound 1\u2009a and comparable to various chloride\u2010bridged complexes or other bridges leading to superexchange pathways in bimetallic gadolinium complexes. was synthesized according to literature known procedure.All experiments were performed under inert gas conditions in N2Li(NtBu)2S(tBuN)2LnCl2}2General synthesis for [{(thf)\u20092] 1\u2009a\u22c5\u2009ClLi(thf)\u2013e: A mixture of [(thf)4Li2(NtBu)4S] and LnCl3 (0.8080\u2005mmol) is dissolved in thf (20\u2005mL) at ambient temperature. After stirring for 1 d the reaction mixture is concentrated under reduced pressure (7\u2005mL), filtered and subsequently, the solvent is removed under reduced pressure. The residue is dissolved in toluene (5\u2005mL) and the solution is filtered. Crystal growth of 1\u2009a\u2013e starts within several minutes to hours at ambient temperature. For a complete crystallization, the mixture is stored at \u221234\u2009\u00b0C yielding colorless crystals suitable for X\u2010ray analysis after several days. The solvent is removed, and the crystalline product is washed with n\u2010pentane (2\u00d72\u2005mL). 1\u2009a: Yield: 266.9\u2005mg (40\u2009%); elemental analysis calcd (%) for C56H120Cl5Gd2Li3N8O6S2(C7H8): C 45.30, H 7.72, N 6.71, S 3.84; found: C 42.91, H 6.92, N 6.51, S 4.47. 1\u2009b: Yield: 304.6\u2005mg (45\u2009%); elemental analysis calcd (%) for C56H120Cl5Tb2Li3N8O6 S2(C7H8): C 45.21, H 7.71, N 6.69, S 3.84; found: C 43.64, H 7.87, N 6.88, S 4.80. 1\u2009c: Yield: 281.5\u2005mg (41\u2009%); elemental analysis calcd (%) for C56H120Cl5Dy2Li3N8O6S2(C7H8): C 45.02, H 7.68, N 6.67, S 3.91; found: C 42.63, H 7.62, N 6.61, S 4.14. 1\u2009d: Yield: 137.1\u2005mg (20\u2009%); elemental analysis calcd (%) for C56H120Cl5Ho2Li3N8O6S2(C7H8): C 44.89, H 7.65, N 6.65, S 3.80; found: C 41.27, H 7.06, N 6.71, S 3.92. 1\u2009e: Yield: 113.8\u2005mg (17\u2009%); elemental analysis calcd (%) for C56H120Cl5Er2Li3N8O6S2(C7H8): C 44.76, H 7.60, N 6.63, S 3.79; found: C 42.67, H 7.60, N 7.14, S 5.08.2Li(NtBu)2S(tBuN)2LnCl2(thf)2] 2\u2009aGeneral synthesis for [{(thf)\u2013e: [{(thf)2Li(NtBu)2S(tBuN)2LnCl2}2\u2009\u22c5\u2009ClLi(thf)2] (1\u2009a\u2013e) (150.0\u2005mg) is dissolved in thf (3\u2005mL) and filtered. Crystallization of 2\u2009a\u2013e starts within hours at ambient temperature where upon the mixture is stored at \u221234\u2009\u00b0C to improve the yield. The target compound is isolated and washed with n\u2010pentane (2\u00d71\u2005mL) yielding crystals suitable for X\u2010ray analysis. 2\u2009a: color: colorless; Yield: 98.3\u2005mg (65\u2009%); elemental analysis calcd (%) for C32H68Cl2GdLiN4O4S: C 45.75, H 8.16, N 6.67, S 3.82; found: C 45.82, H 8.59, N 6.54, S 4.01. 2\u2009b: color: colorless; Yield: 99.6\u2005mg (66\u2009%); elemental analysis calcd (%) for C32H68Cl2TbLiN4O4S: C 45.66, H 8.14, N 6.66, S 3.81; found: C 45.98, H 8.66, N 6.50, S 4.34. 2\u2009c: color: colorless; Yield: 105.2\u2005mg (70\u2009%); elemental analysis calcd (%) for C32H68Cl2DyLiN4O4S: C 45.47, H 8.11, N 6.63, S 3.79; found: C 46.58, H 8.71, N 6.44, S 4.77. 2\u2009d: color: pale orange; Yield: 115.5\u2005mg (77\u2009%); elemental analysis calcd (%) for C32H68Cl2HoLiN4O4S: C 45.34, H 8.09, N 6.61, S 3.78; found: C 45.89, H 8.50, N 6.38, S 3.90. 2\u2009e: color: light pink; here, only 100.0\u2005mg of 2\u2009e were used (1\u2009e); Yield: 74.6\u2005mg (74\u2009%); elemental analysis calcd (%) for C32H68Cl2ErLiN4O4S: C 45.21, H 8.06, N 6.59, S 3.77; found: C 45.55, H 8.61, N 6.35, S 4.13.Crystallographic data: Single crystals were selected under cooling using the X\u2010Temp2 device.2 using SHELXL1\u2009a); 2069110 (1\u2009b), 2069111 (1\u2009c), 2069112 (1\u2009d), 2069113 (1\u2009e), 2069114 (2\u2009a); 2069115 (2\u2009b), 2069116 (2\u2009c), 2069117 (2\u2009d), 2069118 (2\u2009e), 2069119 (3\u2009b)2069109 2Li(NtBu)2S(tBuN)2LnCl2}2\u2009\u22c5\u2009ClLi(thf)2] 1\u2009a\u2013e, with a: Ln=Gd, b: Tb, c: Dy, d: Ho, e: Er and species [{(thf)2Li(NtBu)2S (tBuN)2LnCl2(thf)2] 2\u2009a\u2013e were prepared by loading crushed crystalline samples into tubes in an argon glove\u2010box. Sufficient liquid eicosane (at 60\u2009\u00b0C) was added to saturate and cover the samples to prevent crystallite torquing and provide good thermal contact between the sample and the bath. Tubes were sealed air\u2010tight before transferred to the magnetometer. Magnetic susceptibility measurements were collected using a Quantum Design MPMSXL SQUID magnetometer and Quantum Design MPMS3 SQUID magnetometer, respectively. DC susceptibility data measurements were performed at temperatures ranging from 2 to 300\u2005K for 1\u2009a\u2013e and 2\u2009b\u2013e, using applied fields of 0.1, 0.5 and 1\u2005T. Ac magnetic susceptibility data measurements were performed using a 3.6 (MPMSXL) and 4\u2005Oe (MPMS3) switching field, respectively. All data were corrected for diamagnetic contributions from the eicosane and core diamagnetism estimated using Pascal's constants.\u03c7\u2032 and \u03c7\u2032\u2032 in terms of frequency, constant temperature susceptibility (\u03c7T), adiabatic susceptibility (\u03c7S), relaxation time (\u03c4), and a variable representing the distribution of relaxation times (\u03b1).The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file."} +{"text": "Cell Discovery (2021) 7:103Correction to: 10.1038/s41421-021-00341-7, Published online 31 October 20211, we missed some errors while proofreading the manuscript for the publication. The statistical values in Abstract and Results sections were inconsistent with those in Table FMR1NB, Pmeta\u2009=\u20097.61\u2009\u00d7\u200910\u22129, OR\u2009=\u20091.31; rs7259428, 19q12, ZNF536, Pmeta\u2009=\u20098.20\u2009\u00d7\u200910\u221215, OR\u2009=\u20090.65) showing consistent association with male sexual orientation.\u201d On page 2, the second column, the lines 28\u201335 should be \u201cAll four SNPs showed consistent association in the independent validation samples, and two SNPs reached a genome-wide significance level (P\u2009<\u20095\u2009\u00d7\u200910\u22128) after meta-analysis, including SNP rs7259428 on chromosome 19q12 (located in ZNF536, P\u2009=\u20098.20\u2009\u00d7\u200910\u221215, OR\u2009=\u20090.65; Fig. 1e and Table FMR1NB, P\u2009=\u20097.61\u2009\u00d7\u200910\u22129, OR\u2009=\u20091.31; Fig. 1d and Table ZNF356\u2019 should be \u2018ZNF536\u2019. The corrected version of the Table In the initial published version of this article"} +{"text": "Four copper(II) cations are situated in a distorted square-pyramidal environment, while two copper(II) cations are located in a slightly square-planar geometry. Three of the copper(II) cations occupy three vertices of an open cubane Cu3O4.In the title Schiff base hexa\u00adnuclear copper(II) complex, two discrete environments are present in the structure: CuNO 3-1,3-bis\u00ad{[1-(2-oxidophen\u00adyl)ethyl\u00adidene]amino}\u00adpropan-2-olato)-\u03bc3-hydroxido-dinitrato\u00adhexa\u00adcopper(II) ethanol tris\u00adolvate, [Cu6(C19H19N2O3)3(NO3)2(OH)(H2O)2]\u00b73C2H5OH, corres\u00adponds to a non-symmetric hexa\u00adnuclear copper complex. The complex exhibits one core in which three CuII metal centres are mutually inter\u00adconnected, two by two, via three phenolato oxygen anions acting in a \u03bc2-mode. These three copper cations are inter\u00adconnected in a \u03bc3-mode by one hydroxyl group. An open-cube structure is generated in which each of the CuII cations of the three CuO4N units is connected by two \u03bc2-O anions from phenolate groups and one \u03bc3-O atom from a hy\u00addroxy anion. Each of the three penta\u00adcoordinated CuII cations situated in the open-cube unit has a distorted NO4 square-pyramidal environment. Each of these three CuII centres is inter\u00adconnected with another CuII cation via one enolate O atom in \u03bc2-mode, yielding one CuNO4 unit and two CuNO3 units. The penta\u00adcoordinated CuII atom has a distorted square-pyramidal environment while the two tetra\u00adcoordinated copper(II) cations are situated in a square-planar environment. A series of intra\u00admolecular O\u2014H\u22efO hydrogen bonds are observed. In the crystal, the units are connected two by two by inter\u00admolecular C\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, thus forming sheets parallel to the ac plane.The title mol\u00adecular structure, namely, di\u00adaqua\u00adtris\u00ad(\u03bc That their structures present symmetrical or asymmetrical pendant arms and bear donor atoms is an asset widely exploited in coordination chemistry. The presence of donor sites on aliphatic or aromatic arms has made it possible to prepare a wide variety of compounds with various structures and inter\u00adesting physical and chemical properties. 1,3-Di\u00adamino\u00adpropan-2-ol, which has three donor sites, is a good precursor for the synthesis of ligands with several cavities that can act as chelating agents and/or as bridging ligands ethyl\u00adidene]}-2-hy\u00addroxy\u00adpro\u00adpane-1,3-di\u00adamine (H3L). The reaction of ligand H3L with copper nitrate yielded a complex in which the ligand reacted in tri-deprotonated form as L3\u2212. The coordination complex is formulated as [Cu6L3(NO3)2(OH)(H2O)2]\u00b73(EtOH) (I)II cations. The two imino nitro\u00adgen atoms of the ligand are coordinated to two different Cu cations. One of the phenolato O atoms bridges two copper cations, while the second phenolato O atom is coordinated to a third copper cation. The third copper cation is bridged to the central copper cation via the enolato oxygen anion. The tri-deprotonated ligand coordinates in a hepta\u00addentate mode , thus forming four fused chelate rings (two five-membered and two six-membered). Two discrete environments are observed in the structure: CuNO4 and CuNO3. The coordination environments for Cu1, Cu3, Cu5 and Cu6 are best described as square-pyramidal, as shown by the Addison \u03c4 parameter calculated from the largest angles (Table\u00a01W) of the basal coordination plane are almost coplanar and the Cu6 cation is displaced toward the apical atom (O201) by 0.0963\u2005(9)\u2005\u00c5. The cissoid angles are in the range 86.12\u2005(9)\u201394.66\u2005(9)\u00b0 while the transoid angles are 171.23\u2005(9) and 174.18\u2005(9)\u00b0. In the basal plane, the Cu6\u2014N6 [1.942\u2005(2)\u2005\u00c5] and the Cu6\u2014Oligand distances [1.935\u2005(2) and 1.863\u2005(2)\u2005\u00c5] are shorter than the distance of Cu6\u2014O2W [2.028\u2005(2)\u2005\u00c5]. The distance between the copper and the nitrato oxygen anion [Cu6\u2014O14B = 2.45\u2005(2)\u2005\u00c5] in the apical position is longer than the distances to the atoms in the equatorial plane because of Jahn\u2013Teller distortion, which is typical for copper(II) d9 atoms ethanone and 1,3-di\u00adamino\u00adpropan-2-ol in a 2:1 ratio in ethanol yielded the ligand s Table\u00a01 around C3O4 open cube, the basal planes are occupied by one imino nitro\u00adgen atom, one phenolate oxygen anion, one enolato oxygen anion from the same ligand mol\u00adecule and the O atom of the hy\u00addroxy oxygen anion that connects the three copper cations. The copper cations situated on the corners of the open cube are connected by two \u03bc2-Ophenolato and one \u03bc3-Ohy\u00addroxy atoms. In each case, the apical position is occupied by one phenolate oxygen anion from another ligand. The donor atoms of the basal coordination planes of Cu1, Cu3 and Cu5 centres are situated almost in the same plane and the copper cations are displaced from the corresponding apical positions and O\u2014Cu\u2014O [72.34\u2005(7)\u201386.17\u2005(8)\u00b0] angles, which deviate severely from the ideal value of 90\u00b0 expected for a perfect cube. The atoms defining the three sides of the open cube are almost coplanar and are irregular with edges of different lengths, i.e. for Cu1/O1/Cu5/O10 these are O1\u2014Cu1 = 1.877\u2005(2)\u2005\u00c5, O10\u2014Cu1 = 2.004\u2005(2)\u2005\u00c5, O1\u2014Cu5 = 2.453\u2005(2)\u2005\u00c5 and O10\u2014Cu5 = 1.978\u2005(2)\u2005\u00c5. Additionally, the dihedral angles values of 78.11\u2005(6), 75.77\u2005(5) and 77.57\u2005(5)\u00b0 between the sides, two by two, confirm the distortion of the open cube. The bond lengths involving the bridging phenolate oxygen anions and the copper cations are asymmetrical: O1\u2014Cu1 = 1.877\u2005(2)\u2005\u00c5 and O1\u2014Cu5 = 2. 453\u2005(2)\u2005\u00c5; O4\u2014Cu1 = 2.389\u2005(2)\u2005\u00c5 and O4\u2014Cu3 = 1.896\u2005(2); and O7\u2014Cu5 = 1.889\u2005(2)\u2005\u00c5 and O7\u2014Cu3 = 2.365\u2005(2)\u2005\u00c5. The distances of the \u03bc3-bridging O atom to the copper cations are slightly different: O10\u2014Cu1 = 2.005\u2005(2)\u2005\u00c5, O10\u2014Cu5 = 1.978\u2005(2)\u2005\u00c5 and O10\u2014Cu3 = 2.004\u2005(2)\u2005\u00c5. The axial bond lengths are longer than the equatorial bond lengths as a result of the Jahn\u2013Teller distortion . The three copper cations are placed at the vertices of an almost isosceles triangle with distances values of 3.1801\u2005(4)\u2005\u00c5 (Cu1\u2014Cu5), 3.1823\u2005(4)\u2005\u00c5 (Cu3\u2014Cu5) and 3.2140\u2005(5)\u2005\u00c5 (Cu1\u2014Cu3) and angle values of 60.68\u2005(1)\u00b0 (Cu1\u2014Cu5\u2014Cu3), 59.69\u2005(1)\u00b0 (Cu5\u2014Cu1\u2014Cu3) and 59.62\u2005(1)\u00b0 (Cu1\u2014Cu3\u2014Cu5).For Cu1, Cu3 and Cu5, which are situated on the vertices of the CuW/O6. The \u03c44 \u201394.26\u2005(10)\u00b0] and the transoid angles are 171.59\u2005(9) and 174.77\u2005(10)\u00b0. The Cu4 cation is coordinated by one enolato oxygen anion (O5), one phenoxo oxygen anion (O6), one azomethine nitro\u00adgen atom (N4) of the ligand, and one O atom from a coordinated water mol\u00adecule. The distances of Cu4 to the coordinated atoms from the ligand are comparable with those involving Cu2. The Cu4\u2014O1W distance value of 1.961\u2005(2)\u2005\u00c5 is similar to those reported for square-planar copper(II) complexes \u201395.34\u2005(9)\u00b0 and the transoid angles are 171.10\u2005(9) and 173.69\u2005(9)\u00b0. The double-bond character of the C\u2014N bonds [overall values 1.286\u2005(3)\u20131.295\u2005(3)\u2005\u00c5] is indicative of the presence of the imino groups in the three ligands.For the Cu2 and Cu4 centres, the coordination environments can be best described as slightly distorted square planar with r.m.s. deviations from planarity of 0.0601\u2005\u00c5 for Cu2/O2/N2/O3/O11 and 0.0909\u2005\u00c5 for Cu4/N4/O5/O1water\u2014H\u22efOethanol and Owater\u2014H\u22efOnitrate) and C\u2014H\u22efO (Cphenolate\u2014H\u22efOnitrate) Fig.\u00a02. The comne Fig.\u00a03. The cooN,N\u2032-bis\u00ad[(1-(2-hy\u00addroxy\u00adphen\u00adyl)ethyl\u00adidene)]-2-hy\u00addroxy\u00adpropane-1,3-di\u00adamine has been widely used in coordination chemistry. The current release of the CSD ethyl\u00adidene]}-2-hy\u00addroxy\u00adpropane-1,3-di\u00adamine (HL3), which was prepared according to a literature method : 3538 (OH), 3268 (OH), 1605 (C=N), 1538 (C=C), 1528 (C=C), 1455 (C=C), 1247 (C\u2014O), 1043, 760. Analysis calculated for C19H22N2O3: C, 69.92; H, 6.79; N, 8.58. Found: C, 69.90; H, 6.76; N, 8.56%. A solution of Cu(NO3)2\u00b73H2O in 5\u2005mL of ethanol was added to a solution of H3L in 10\u2005mL of ethanol at room temperature. The initial yellow solution immediately turned dark green and was stirred for 30\u2005min. The mixture was filtered, and the filtrate was kept at 298\u2005K. After one week, light-green crystals suitable for X-ray diffraction were collected and formulated as [Cu6L3(NO3)2(OH)(H2O)2]\u00b73EtOH. FT\u2013IR : 1625, 1600, 1540, 1446, 1382, 1304, 1258, 1180, 1120, 1007, 895, 760. Analysis calculated for C63H80Cu6N8O21: C, 45.40; H, 4.84; N, 6.72. Found: C, 45.38; H, 4.82; N, 6.74%.Reaction of 1-(2-hy\u00addroxy\u00adphen\u00adyl)ethanone and 2-hy\u00addroxy\u00adpropane-1,3-di\u00adamine in a 2:1 ratio in ethanol yielded the ligand 2, CH3 groups, hydroxyl groups of ethanol mol\u00adecules and water mol\u00adecules) were geometrically optimized and refined as riding with Uiso(H) = 1.2Ueq(C) (1.5 for CH3 and OH groups).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021005570/ex2045sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021005570/ex2045Isup2.hklStructure factors: contains datablock(s) I. DOI: 2086932CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of the title compound comprises two independent mol\u00adecules that mainly differ in the orientation of the phenyl ring to the rest of the mol\u00adecule. 16H13N3OS, comprises two mol\u00adecules (A and B) with similar conformations that differ mainly in the orientation of the phenyl group relative to the rest of the mol\u00adecule, as expressed by the Cthio\u00adamide\u2014Nthio\u00adamide\u2014Cphen\u00adyl\u2014Cphen\u00adyl torsion angle of 49.3\u2005(3)\u00b0 for mol\u00adecule A and of 5.4\u2005(3)\u00b0 for mol\u00adecule B. In the crystal, two inter\u00admolecular N\u2014H\u22efN hydrogen bonds lead to the formation of a dimer with R22(10) graph-set notation. A Hirshfeld surface analysis revealed that H\u22efH inter\u00adactions are the most important inter\u00admolecular inter\u00adactions, contributing 40.9% to the Hirshfeld surface.The asymmetric unit of the title compound, C The presence of bifunctional properties in thio\u00adamides, resulting from the presence of nitro\u00adgen and sulfur atoms, and their participation in reactions as electrophilic or nucleophilic reagents can lead to the formation of different heterocyclic compounds. Several review articles have been published on the syntheses, physico-chemical properties and applications of thio\u00adamides and 49.3\u2005(3)\u00b0, respectively, in mol\u00adecule A and 83.78\u2005(19) and 5.4\u2005(3)\u00b0 in mol\u00adecule B. As a result, there are differences in the intra\u00admolecular distances between the sulfur and hydrogen atoms in mol\u00adecules A and B. In mol\u00adecule A, the contacts S1A\u22efH9AB and S1A\u22efH13A are 2.873 and 2.897\u2005\u00c5 whereas the corresponding distances in mol\u00adecule B are 3.054 and 2.578\u2005\u00c5. The phenyl and pyrimidine rings in both mol\u00adecules are essentially coplanar, with r.m.s. deviations of 0.0225 and 0.0119\u2005\u00c5 for mol\u00adecule A and B, respectively. Fig.\u00a02The title compound crystallizes with two mol\u00adecules, it Fig.\u00a01. In mol\u00adA and B form a dimer with an A inter\u00adacts with mol\u00adecule B by a C\u2014H\u22ef \u03c0 inter\u00adaction . Other weak C7A\u2014H7A\u22efO1B, C7A\u2014H7A \u22efO1B, C7B\u2014H7B\u22efO1A, C9A\u2014H9AB\u22efS1A and C13B\u2014H13B\u22efS1B hydrogen bonds link adjacent dimers, forming supra\u00admolecular layers expanding parallel to (010) Fig.\u00a04. The ovell Fig.\u00a05.CrystalExplorer17.5 whereby H\u22efH contacts are responsible for the largest contribution (40.9%) to the Hirshfeld surface. C\u22efH/H\u22efC contribute 23.7%, S\u22efH/H\u22efS contacts 10.7%, N\u22efH/H\u22efN contacts 8.1% and O\u22efH/H\u22efO contacts 7.0% to the total Hirshfeld surface. The contributions of further contacts are only minor and amount to C\u22efC (4.0%), S\u22efC/C\u22efS (1.9%), N\u22efC/C\u22efN (1.2%), S\u22efS (1.0%), S\u22efC/C\u22efS (0.6%), O\u22efN/N\u22efO (0.2%) and O\u22efC/C\u22efO (0.1%).A Hirshfeld surface (HS) analysis -one moiety with a similar planar conformation as that in the title structure: AFOCIJ and 4\u2005ml of dimethyl sulfoxide were injected into a round-bottomed flask with a volume of 100\u2005ml. Then the reaction flask was heated to 403\u2005K for 6\u2005h. After the end of the reaction, the flask was cooled and 40\u2005ml of an aqueous sodium hydroxide solution were added. The resulting mixture was filtered, then added to a dilute solution of sulfuric acid (pH 6). The formed precipitate was filtered off and recrystallized in methanol. In total, 0.5\u2005g (64.0%) of the product were obtained, m.p. 481\u2013483\u2005K.0.435\u2005g (0.0025\u2005mol) of 2,3-di\u00admethyl\u00adquinazoline-4-one, 0.465\u2005g (0.005\u2005mol) of aniline, 0.24\u2005g (0.0075\u2005mol) of sulfur, 0.05\u2005g of sodium sulfide (NaUiso(H) = 1.5Ueq(Cmeth\u00adyl) and 1.2Ueq(C), respectively. H atoms bonded to nitro\u00adgen were located in a difference-Fourier map, and their positional and isotropic displacement parameters were freely refined.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021013116/wm5629sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021013116/wm5629Isup3.cmlSupporting information file. DOI: 10.1107/S2056989021013116/wm5629Isup3.hklStructure factors: contains datablock(s) I. DOI: 2127513CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Two similar mol\u00adecules make up the asymmetric unit of the title compound. The crystal structure features short C\u2014H\u22efCl and C\u2014H\u22efO contacts and C\u2014H\u22ef\u03c0 and van der Waals inter\u00adactions. 16H14Cl2N2O, comprises two similar mol\u00adecules, A and B, in which the dihedral angles between the two aromatic rings are 70.1\u2005(3) and 73.2\u2005(2)\u00b0, respectively. The crystal structure features short C\u2014H\u22efCl and C\u2014H\u22efO contacts and C\u2014H\u22ef\u03c0 and van der Waals inter\u00adactions. The title compound was refined as a two-component non-merohedral twin, BASF 0.1076\u2005(5). The Hirshfeld surface analysis and two-dimensional fingerprint plots show that H\u22efH , Cl\u22efH/H\u22efCl and C\u22efH/H\u22efC inter\u00adactions are the most important contributors to the crystal packing.The asymmetric unit of the title compound, C In mol\u00adecule A, the N2/N1/C2/C1/Cl1/Cl2 moiety is approximately planar, with a maximum deviation of 0.110\u2005(2)\u2005\u00c5, and makes dihedral angles of 1.2\u2005(2) and 71.3\u2005(2)\u00b0, respectively, with the C3\u2013C8 and C10\u2013C15 rings. In mol\u00adecule B, the N4/N3/C18/C17/Cl3/Cl4 moiety is approximately planar with a maximum deviation of 0.046\u2005(6)\u2005\u00c5, and makes dihedral angles of 9.57\u2005(18) and 75.94\u2005(19)\u00b0, respectively, with the C19\u2013C24 and C26\u2013C31 rings.There are two comparable mol\u00adecules A (with Cl1) and B (with Cl3) in the asymmetric unit of the title compound Fig.\u00a01. The dihvia C\u2014H\u22efCl short contacts, yielding supra\u00admolecular chains along the b-axis direction. Adjacent chains are linked by C\u2014H\u22efO contacts, generating a two-dimensional array parallel to the bc plane . The Cl\u22efH/H\u22efCl inter\u00adactions appear as two symmetrical broad wings with de + di = 2.70\u2005\u00c5 and contribute 24.6% to the Hirshfeld surface for mol\u00adecule A, and with de + di = 2.70\u2005\u00c5 and contribute 26.7% to the Hirshfeld surface for mol\u00adecule B . The pair of characteristic wings in the fingerprint plot delineated into H\u22efC/C\u22efH contacts have the tips at de + di = 2.80\u2005\u00c5 for mol\u00adecule A and at de + di = 2.85\u2005\u00c5 for mol\u00adecule B. The remaining contributions for both mol\u00adecules A and B are from N\u22efH/H\u22efN, O\u22efH/H\u22efO, N\u22efC/C\u22efN, Cl\u22efO/O\u22efCl, Cl\u22efC/C\u22efCl, C\u22efC, Cl\u22efN/N\u22efCl, O\u22efC/C\u22efO and Cl\u22efCl contacts, which are less than 4.6% and have a negligible effect on the packing. The percentage contributions of all inter\u00adactions are listed in Table\u00a03A and B can be attributed to the different mol\u00adecular environments of the A and B mol\u00adecules in the crystal structure.The overall two-dimensional fingerprint plot and those delineated into H\u22efH, Cl\u22efH/H\u22efCl and C\u22efH/H\u22efC contacts in mol\u00adecules ng Fig.\u00a06b. The C B Fig.\u00a06c. The pet al., 2016E)-1--2-phenyl\u00addiazene unit resulted in 27 hits. Eight compounds are closely related to the title compound, viz. 4-{2,2-di\u00adchloro-1-[diazen\u00adyl]ethen\u00adyl}-N,N-di\u00admethyl\u00adaniline \u2005\u00c5]. In DULTAI, the dihedral angle between the two aromatic rings is 64.12\u2005(14)\u00b0. The crystal structure is stabilized by a short C\u2014H\u22efCl contact, C\u2014Cl\u22ef\u03c0 and van der Waals inter\u00adactions. In HONBOE and HONBUK, the aromatic rings form dihedral angles of 60.9\u2005(2) and 64.1\u2005(2)\u00b0, respectively. In the crystals, mol\u00adecules are linked through weak X\u22efCl contacts (X = Br for HONBOE and Cl for HONBUK), C\u2014H\u22efCl and C\u2014Cl\u22ef\u03c0 inter\u00adactions into sheets parallel to the ab plane. Additional van der Waals inter\u00adactions consolidate the three-dimensional packing. In HODQAV, the benzene rings make a dihedral angle of 56.13\u2005(13)\u00b0. Mol\u00adecules are stacked in columns along the a-axis direction via weak C\u2014H\u22efCl hydrogen bonds and face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions. The crystal packing is further consolidated by short Cl\u22efCl contacts. In XIZREG, the benzene rings form a dihedral angle of 63.29\u2005(8)\u00b0 and the mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds into zigzag chains running along the c-axis direction. The crystal packing also features C\u2014Cl\u22ef\u03c0, C\u2014F\u22ef\u03c0 and N\u2014O\u22ef\u03c0 inter\u00adactions. In the crystals of LEQXIR and LEQXOX, the dihedral angles between the aromatic rings are 56.18\u2005(12) and 60.31\u2005(14)\u00b0, respectively. In LEQXIR, C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and short C\u2014Cl\u22efO contacts occur and in LEQXOX, C\u2014H\u22efN and short Cl\u22efCl contacts are observed.In GUPHIL, the benzene rings subtend a dihedral angle of 77.07\u2005(10)\u00b0. In the crystal, mol\u00adecules are associated into inversion dimers et al., 2021et al., 2018Z)-1-(4-meth\u00adoxy\u00adphen\u00adyl)-2-(4-methyl\u00adbenzyl\u00adid\u00adene)hydrazine , tetra\u00admethyl\u00adethylenedi\u00adamine , CuCl and CCl4 . After 1\u20133\u2005h , the reaction mixture was poured into a 0.01\u2005M solution of HCl , and extracted with di\u00adchloro\u00admethane (3 \u00d7 20\u2005mL). The combined organic phase was washed with water (3 \u00d7 50\u2005mL) and brine (30\u2005mL), dried over anhydrous Na2SO4 and concentrated in vacuo by a rotary evaporator. The residue was purified by column chromatography on silica gel using appropriate mixtures of hexane and di\u00adchloro\u00admethane (3/1\u20131/1). Crystals suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution. Colorless solid (65%); m.p. 355\u2005K. Analysis calculated for C16H14Cl2N2O: C 59.83, H 4.39, N 8.72; found: C 59.78, H 4.32, N 8.69%. 1H NMR \u03b4 7.79 , 7.26 , 7.10 , 6.95 , 3.88 , 2.42 . 13C NMR \u03b4 162.48, 148.12, 147.82, 138.47, 129.90, 129.76, 129.41, 128.85, 125.23, 114.14, 55.58 and 21.48. ESI\u2013MS: m/z: 322.14 [M\u00a0+\u00a0H]+.The title compound was synthesized according to a reported method (Mukhtarova Uiso(H) = 1.2 or 1.5Ueq(C). Owing to poor agreement between observed and calculated intensities, eight outliers I. DOI: 10.1107/S2056989021008756/zn2009Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021008756/zn2009Isup3.cmlSupporting information file. DOI: 1984582CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Cellular stress has been associated with inflammation, yet precise underlying mechanisms remain elusive. In this study, various unrelated stress inducers were employed to screen for sensors linking altered cellular homeostasis and inflammation. We identified the intracellular pattern recognition receptors NOD1/2, which sense bacterial peptidoglycans, as general stress sensors detecting perturbations of cellular homeostasis. NOD1/2 activation upon such perturbations required generation of the endogenous metabolite sphingosine\u20101\u2010phosphate (S1P). Unlike peptidoglycan sensing via the leucine\u2010rich repeats domain, cytosolic S1P directly bound to the nucleotide binding domains of NOD1/2, triggering NF\u2010\u03baB activation and inflammatory responses. In sum, we unveiled a hitherto unknown role of NOD1/2 in surveillance of cellular homeostasis through sensing of the cytosolic metabolite S1P. We propose S1P, an endogenous metabolite, as a novel NOD1/2 activator and NOD1/2 as molecular hubs integrating bacterial and metabolic cues. Intracellular pattern recognition receptors NOD1/2, best known as sensor of bacterial peptidoglycans, can also be directly activated by signaling lipid sphingosine\u20101\u2010phosphate accumulating upon perturbed cellular homeostasis, thus linking general stress to inflammatory responses. Altogether, we established that sphingolipid metabolism governs inflammation triggered by cellular stress and identified a central role for S1P in triggering NOD1/2\u2010dependent inflammation upon stress induction.IL6 and IL8 expression and production correlated with NOD1/2 expression, without considerable differences of cell death in association with NOD1 or NOD2 expression upon perturbation of cellular homeostasis were compromised in both Nod1/2 dKO and Rip2 KO cells and NOD1/2 double knockout (dKO). NF\u2010\u03baB luciferase assays confirmed that perturbation of cellular homeostasis induced NF\u2010\u03baB activation, while NF\u2010\u03baB activation and dKO Fig\u00a0. Consistlls Figs . AccordiMDMs Fig\u00a0C and D. et al,et al,et al,ACER1/2, SMPD3,SPHK1 and other key enzymes involved in sphingolipid metabolism encode the sphingosine kinase resulted in lower IL8 production upon Shigella infection, suggesting the involvement of S1P in Shigella\u2010induced NF\u2010\u03baB activation were employed for two reasons: (i) Sphingolipid metabolism in cancer cells is dysregulated or neutral sphingomyelinase (GW4869), uncovering that S1P is mainly produced via the hydrolysis pathway upon ER stress , S1PR2 (JTE013), S1PR1 and S1PR3 (VPC23019), and S1PR4 (CYM50358), as well as the general S1PR inhibitor FTY720 or blocking GPCR signaling by pertussis toxin, were employed. All inhibitors failed to block IL6 production by cytosolic S1P with the S1P molecule using AutoDock domain of human NOD1 is critical for the interaction with S1P or caspase recruitment domain (CARD)\u2010containing proteins, resembling inflammasomes in vertebrates and B6N.129S6\u2010Sphk2tm1Rlp/J (Sphk2\u2212/\u2212) mice were originally obtained from the Jackson Laboratory . These mice were maintained under specific pathogen\u2010free (SPF) conditions at the Max Planck Institute for Infection Biology in Berlin, Germany. 6\u2010 to 15\u2010week\u2010old female mice were employed for the experiments.B6N.129S6\u2010Sphk1et al,NLRP3\u2010GFP was obtained from Addgene. NOD1\u2010 or NOD2\u2010related plasmids were constructed as described , anti\u2010GFP , anti\u2010GFP , anti\u2010RIP2 , anti\u2010SPHK2 , anti\u2010phospho\u2010p38 , anti\u2010phospho\u2010JNK , anti\u2010phospho\u2010ERK1/2 , anti\u2010phospho\u2010p65 , and anti\u2010NOD1 .l\u2010glutamine , 10\u00a0mM HEPES buffer , pH 7.2\u20107.5, and 50\u00a0\u00b5M 2\u2010mercaptoethanol . To differentiate THP\u20101 into macrophage\u2010like cells, THP\u20101 cells were stimulated with 50\u00a0ng/ml phorbol 12\u2010myristate 13\u2010acetate for 24\u00a0h and then incubated with fresh medium for another 48\u00a0h. HEK293T WT and HEK293T NOD1/2 dKO cells were generated as described were obtained from tibial and femural bones and generated with DMEM containing 20% L929 cell supernatant, 10% FBS, 5% heat\u2010inactivated horse serum, 1\u00a0mM sodium pyruvate, and 2\u00a0mM l\u2010glutamine. For BMDM stimulation experiments, BMDMs were treated with IFN\u2010\u03b3 (20\u00a0ng/ml) overnight before stimulation.The human monocytic cell line THP\u20101 was obtained from the American Type Culture Collection and maintained in RPMI 1640 with 10% (v:v) heat\u2010inactivated fetal bovine serum , 1\u00a0mM sodium pyruvate , 2\u00a0mM ueous one solution reagent (Promega) according to the manufacturer\u2019s instructions. Cell viability upon various stimulations was normalized to corresponding controls.Cell viability assays were performed with CellTiter 96 AQShigella flexneri M90T afaE .NOD1/2 KO NF\u2010\u03baB reporter cells were seeded in 96\u2010well plates and stimulated with various stressors for 8\u00a0h. Then, cells were lysed in lysis buffer (Thermo Fisher Scientific) and luciferase activities were determined with firefly luciferase glow kit (Thermo Fisher Scientific) according to the manufacturer\u2019s instructions. Luciferase activities were normalized to the amount of protein determined with Coomassie Plus kit (Thermo Fisher Scientific). Fold inductions were calculated against normalized luciferase activities of corresponding controls.HEK293T WT and 6 cells were treated with indicated stimuli for 2\u00a0h. After washing twice with Dulbecco's phosphate\u2010buffered saline (DPBS) without calcium and magnesium, cells were collected and suspended in 300\u00a0\u00b5l of DPBS. Afterward, cells were transferred into 1.5\u2010ml Eppendorf tubes precooled at \u221280\u00b0C. The lipidomic analysis was performed by Lipotype GmbH. For analysis of sphingosine\u2010related metabolites, 3\u00a0\u00d7\u00a0106 cells were seeded in Lumox 50 dishes and treated with indicated stimuli for 2\u00a0h. After washing twice with 0.9% NaCl preconditioned at 37\u00b0C, the membrane of Lumox dishes was placed into tubes precooled at \u221280\u00b0C. 600\u00a0\u00b5l of quenching solution (dichloromethane/ethanol) was added into each tube. The measurement was performed by Metanomics Health GmbH.For lipidomic profiling, 1\u00a0\u00d7\u00a0102, 0.1% SDS, 1X PhosSTOP, 1X Protease inhibitor cocktail without EDTA). Protein concentration was measured by BCA assay kit (Thermo Fisher) and diluted with delipidized serum to 1\u00a0\u00b5g/\u00b5l. Then, the diluted samples were added into S1P ELISA plates according to the instructions.Quantification of S1P abundances in cell lysates was performed with S1P ELISA kit according to the manufacturer\u2019s instructions with some modifications. Human neonatal dermal fibroblasts were grown in DMEM full medium. For each condition, 2x T75 flask cells were required. After 4\u2010h stimulation with various stressors, cells were washed with PBS and collected in lysis buffer in a Bio\u2010Plex 200 system according to the manufacturer\u2019s instructions.The mutagenesis was performed with Q5\u00ae site\u2010directed mutagenesis kit according to the manufacturer's instructions. The primers used in this study were as follows: NOD1 E157A F: 5\u2032\u2010GCTGCTGGAGgcgATCTACATGG\u20103\u2032, NOD1 E157A R: 5\u2032\u2010AGCTCCTCCTTCTGGGCA\u20103\u2032, NOD1 D161A F: 5\u2032\u2010GATCTACATGgcgACCATCATGGAGCTGG\u20103\u2032, NOD1 D161A R: 5\u2032\u2010TCCTCCAGCAGCAGCTCC\u20103\u2032, NOD1 D203A F: 5\u2032\u2010CATCCTGGGTgcgGCTGGGGTGG\u20103\u2032, D203A R: 5\u2032\u2010AAGATGGTCTCACCCTGC\u20103\u2032, L218A F: 5\u2032\u2010GCTGCAGAGCgcgTGGGCCACGG\u20103\u2032, L218A R: 5\u2032\u2010CGCTGTAGCAGCATGGAC\u20103\u2032, R237A F: 5\u2032\u2010CTTTCGCTGCgcgATGTTCAGCTG\u20103\u2032, R237A R: 5\u2032\u2010TGGAAGAAGAATTTGACC\u20103\u2032, H257A F: 5\u2032\u2010GCTCTTCAAGgcgTACTGCTACCCAGAGCGG\u20103\u2032, H257A R: 5\u2032\u2010AGGTCCTGCAGACACAGC\u20103\u2032, E267A F: 5\u2032\u2010GGACCCCGAGgcgGTGTTTGCCT\u20103\u2032, E267A R: 5\u2032\u2010CGCTCTGGGTAGCAGTAGTG\u20103\u2032, H290A F: 5\u2032\u2010GGACGAGCTGgcgTCGGACTTGGACC\u20103\u2032, H290A R: 5\u2032\u2010AGGCCATCGAAGGTGAAG\u20103\u2032, E306A F: 5\u2032\u2010CTGCCCCTGGgcgCCTGCCCACC\u20103\u2032, E306A R: 5\u2032\u2010GAGCTGTCAGGCACGCGGC\u20103\u2032, K324A F: 5\u2032\u2010GAAGCTGCTCgcgGGGGCTAGCAAG\u20103\u2032, K324A R: 5\u2032\u2010CCACTGAGCAGGTTGGCC\u20103\u2032, K328A F: 5\u2032\u2010GGGGGCTAGCgcgCTGCTCACAG\u20103\u2032, K328A R: 5\u2032\u2010TTGAGCAGCTTCCCACTG\u20103\u2032, R340A F: 5\u2032\u2010CGAGGTCCCGgcgCAGTTCCTGCGGAAG\u20103\u2032, R340A R: 5\u2032\u2010ATGCCTGTGCGGGCTGTG\u20103\u2032, R344A F: 5\u2032\u2010CCAGTTCCTGgcgAAGAAGGTGCTTCTCCGG\u20103\u2032, R344A R: 5\u2032\u2010CGCGGGACCTCGATGCCT\u20103\u2032, R350A F: 5\u2032\u2010GGTGCTTCTCgcgGGCTTCTCCC\u20103\u2032, R350A R: 5\u2032\u2010TTCTTCCGCAGGAACTGG\u20103\u2032, D372A F: 5\u2032\u2010GGCCCTGCAGgcgCGCCTGCTGA\u20103\u2032, D372A R: 5\u2032\u2010CGCTCGGGGAACATCCTC\u20103\u2032, R373A F: 5\u2032\u2010CCTGCAGGACgcgCTGCTGAGCCAGCTG\u20103\u2032, R373A R: 5\u2032\u2010GCCCGCTCGGGGAACATC\u20103\u2032, R399A F: 5\u2032\u2010GATCATCTTCgcgTGCTTCCAGCACTTCC\u20103\u2032, R399A R: 5\u2032\u2010CAGCAGAAGAGGGGCACA\u20103\u2032, D423A F: 5\u2032\u2010GACCCTGACAgcgGTCTTCCTCCTGGTC\u20103\u2032, D423A R: 5\u2032\u2010ATCGTGCAGTCGGGCAGC\u20103\u2032, R435A F: 5\u2032\u20105\u2032\u2010CCATCTGAACgcgATGCAGCCCAGC\u20103\u2032, R435A R: 5\u2032\u2010ACCTCAGTGACCAGGAGG\u20103\u2032, Q484A F: 5\u2032\u2010GGAGGAGGTGgcgGCCTCCGGGC\u20103\u2032, Q484A R: 5\u2032\u2010TGGGTGAAGACAAAGAGGCTC\u20103\u2032, L495A F: 5\u2032\u2010AGACATGCAGgcgGGCTTCCTGCGG\u20103\u2032, L495A R: 5\u2032\u2010CTCTCCTGCAGCCCGGAG\u20103\u2032, E514A F: 5\u2032\u2010GCAGTCCTATgcgTTTTTCCACCTCACCCTCCAGG\u20103\u2032, E514A R: 5\u2032\u2010TGGTCACCCCCGGGGCCC\u20103\u2032, H517A F: 5\u2032\u2010TGAGTTTTTCgcgCTCACCCTCCAGGCCTTCTTTAC\u20103\u2032, H517A R: 5\u2032\u2010TAGGACTGCTGGTCACCC\u20103\u2032, L540A F: 5\u2032\u2010CACTCAGGAGgcgCTCAGGTTCTTCCAGG\u20103\u2032, L540A R: 5\u2032\u2010CCCACCCTGTCGTCCAGC\u20103\u2032, F544A F: 5\u2032\u2010GCTCAGGTTCgcgCAGGAGTGGATGCCC\u20103\u2032, and F544A R: 5\u2032\u2010AGCTCCTGAGTGCCCACC\u20103\u2032.2+, 1\u00a0mM ATP). To elute the proteins, the agarose was mixed with 2\u00a0ml elution buffer for 1\u00a0min and the eluted proteins were immediately neutralized with 0.2\u00a0ml of 1\u00a0M Tris, pH 10.5.HeLa NOD1 or NOD2 cells from 20 T175 flasks were collected and lysed with 5\u00a0ml lysis buffer . The lysates were incubated with 2\u00a0ml GFP\u2010Trap agarose at 4\u00b0C for 4\u00a0h. The agarose was collected with Poly\u2010Prep columns by gravity flow. Afterward, the agarose was sequentially washed with 50\u00a0ml wash buffer I and 50\u00a0ml wash buffer II . To remove heat shock proteins, columns were further washed with 50\u00a0ml wash buffer III and treated with doxycycline overnight to induce NOD1\u2010GFP or NOD2\u2010GFP expression. Afterward, cells were stimulated with S1P\u2010TAMRA (20\u00a0\u00b5M) in the presence of digitonin (5\u00a0\u00b5g/ml) for 2\u00a0h. Then, cells were fixed with 4% (v:v) paraformaldehyde in PBS, pH 7.4 for 10\u00a0min at room temperature (RT). After washing twice with PBS, cells were incubated with 50\u00a0mM glycine in PBS, pH 7.4 for 10\u00a0min and permeabilized with 0.05% saponin , 1% BSA in PBS for 10\u00a0min. Rabbit anti\u2010GFP antibody (ChromoTek) and Alex Fluor 647\u2010conjugated goad anti\u2010rabbit antibody (Thermo Fisher Scientific) were diluted in PBS and incubated for 1\u00a0h at room temperature. The super resolution imaging was performed with Nanoimager S (Oxford Nanoimaging). For single particle tracking in live cells, Nanoimager S was pre\u2010warmed at 37\u00b0C. HeLa inducible NOD1\u2010GFP or NOD2\u2010GFP cells were stimulated with S1P\u2010TAMRA (20\u00a0\u00b5M) in the presence of digitonin (5\u00a0\u00b5g/ml) for 30\u00a0min. Medium was then replaced with fresh DMEM without phenol red. Live imaging was performed with Nanoimager S at acquisition speed of 100fps.GAPDH), and fold changes were calculated against corresponding controls. qRT\u2013PCR data were representative of at least three independent experiments, with at least 2 technical replicates per experiment. The sequences of primers used in this study were as follows: hGAPDH F: 5\u2032\u2010GGAGCGAGATCCCTCCAAAAT\u20103\u2032, hGAPDH R: 5\u2032\u2010GGCTGTTGTCATACTTCTCATGG\u20103\u2032, hIL6 F: 5\u2032\u2010ACTCACCTCTTCAGAACGAATTG\u20103\u2032, hIL6 R: 5\u2032\u2010CCATCTTTGGAAGGTTCAGGTTG\u20103\u2032, hIL8 F: 5\u2032\u2010TTTTGCCAAGGAGTGCTAAAGA\u20103\u2032, hIL8 R: 5\u2032\u2010AACCCTCTGCACCCAGTTTTC\u20103\u2032, mGAPDH F: 5\u2032\u2010AGGTCGGTGTGAACGGATTTG\u20103\u2032, mGAPDH R: 5\u2032\u2010TGTAGACCATGTAGTTGAGGTCA\u20103\u2032, mIL6 F: 5\u2032\u2010TAGTCCTTCCTACCCCAATTTCC\u20103\u2032, mIL\u20106 F: 5\u2032\u2010TTGGTCCTTAGCCACTCCTTC\u20103\u2032, hS1PR1 F: 5\u2032\u2010TTCCACCGACCCATGTACTAT\u20103\u2032, hS1PR1 R: 5\u2032\u2010GCGAGGAGACTGAACACGG\u20103\u2032, hS1PR2 F: 5\u2032\u2010CTAGCCAGTTCTGAAAGC\u20103\u2032, hS1PR2 R: 5\u2032\u2010ACAGAGGATGACGATGAAG\u20103\u2032, hS1PR3 F: 5\u2032\u2010GAGGAGCCCTTTTTCAAC\u20103\u2032, hS1PR3 R: 5\u2032\u2010TCATTTCAAAGGGAAGCG\u20103\u2032, hS1PR4 F: 5\u2032\u2010GACGCTGGGTCTACTATTGCC\u20103\u2032, hS1PR4 R: 5\u2032\u2010CCTCCCGTAGGAACCACTG\u20103\u2032, hS1PR5 F: 5\u2032\u2010AGGAAGCTCAGTTCACAG\u20103\u2032, and hS1PR5 R: 5\u2032\u2010GATTCTCTAGCACGATGAAG\u20103\u2032.Total RNA was isolated with TRIzol reagent, as described by the manufacturer (Invitrogen). RNA (1\u00a0\u03bcg) was used to generate cDNA via the iScript cDNA Synthesis Kit (Bio\u2010Rad), and qRT\u2013PCR was performed using Power SYBR Green Master Mix (Applied Biosystems) in a StepOne Plus thermocycler (Applied Biosystems). The average threshold cycle (Ct) of quadruplicate reactions was employed for all subsequent calculations using the \u0394\u0394Ct method. Gene expression was normalized to glyceraldehyde\u20103\u2010phosphate dehydrogenase . Data represent fold changes (2\u2212\u0394\u0394CT) in transcripts relative to the appropriate internal control (DMSO). Data were generated from 2 technical replicates and at least three independent experiments. TaqMan probes are listed in Table\u00a0Gene expression was analyzed simultaneously with the 96.96 Dynamic Array Integrated Fluidic Circuits from Fluidigm as previously described following the manufacturer\u2019s protocol using glycogen as co\u2010precipitant. Quality control and quantification of total RNA was analyzed using an Agilent 2100 Bioanalyzer (Agilent Technologies) and a NanoDrop 1000 UV\u2013Vis spectrophotometer (Thermo Fisher Scientific). Microarray experiments were performed as single\u2010color hybridization. Total RNA was amplified and labeled with the Low Input Quick\u2010Amp Labeling Kit (Agilent Technologies). In brief, mRNA was reverse\u2010transcribed and amplified using an oligo\u2010dT\u2010T7 promoter primer and labeled with cyanine 3\u2010CTP. After precipitation, purification, and quantification, 0.75\u00a0\u03bcg labeled cRNA was fragmented and hybridized to custom whole genome human 8\u00a0\u00d7\u00a060K multipack microarrays (Agilent\u2010048908) according to the supplier\u2019s protocol (Agilent Technologies). Scanning of microarrays was performed with 3\u00a0\u03bcm resolution (8x60K) using a G2565CA high\u2010resolution laser microarray scanner (Agilent Technologies). Microarray image data were processed with the Image Analysis/Feature Extraction software G2567AA v. A.11.5.1.1 (Agilent Technologies) using default settings and the GE1_1105_Oct12 extraction protocol. Expression data were analyzed using R scripts. The data were quantile\u2010normalized between arrays, and batch effect of purification state was removed by ComBat overnight at 4\u00b0C. Afterward, agaroses were washed twice with PBS and boiled with sample buffer for SDS\u2013PAGE. For lipid\u2010coated beads IP, lipid\u2010coated beads were incubated with supernatants for 2\u00a0h at room temperature and then washed with lysis buffer for three times and boiled with sample buffer for SDS\u2013PAGE.HeLa NOD1 or NOD2 cells were grown overnight and afterward treated with doxycycline overnight to induce NOD1 or NOD2 expression. Cells of one 10\u2010cm dish were lysed with 500\u00a0\u00b5l of lysis buffer supplemented with protease inhibitors on ice for 20\u00a0min. Lysates were centrifuged at 16,000\u00a0\u00d7\u00a0g for 10\u00a0min, supernatants were heated with SDS sample buffer at 95\u00b0C for 10\u00a0min. Proteins were separated on 4\u201315% SDS gels and transferred onto nitrocellulose membranes. The membranes were incubated with primary antibodies at 4\u00b0C overnight and secondary antibodies at RT for 1\u00a0h, respectively. All antibodies were diluted in PBS supplemented with 0.1% Tween\u201020 and 5% BSA. Membranes were developed with ECL detection Kit and exposed with ChemiDoc (Bio\u2010Rad). The antibody against ACTB/\u03b2\u2010actin was used as loading control.Cells were lysed in RIPA buffer supplemented with protease inhibitor cocktail and PhosSTOP on ice for 10\u00a0min. After centrifugation at 16,000\u00a0\u00d7\u00a0Kd values.Binding of NOD1, NOD2, GFP, or NLRP3\u2010GFP to different ligands was measured by microscale thermophoresis (MST). 20\u00a0nM GFP or GFP\u2010tagged proteins in MST buffer was incubated with different concentrations of ligands. Immediately, samples were loaded into standard glass capillaries (NanoTemper) and thermophoresis analysis was performed on a NanoTemper Monolith NT.115 instrument at 22\u00b0C. A laser on\u2010time of 30\u00a0s and a laser off\u2010time of 5\u00a0s were used. The experiment was performed in triplicates, and the MST curves were fitted using NT analysis software to obtain the et al,et al,et al,et al,AutoDock Tools was satisfied. The system was first minimized with the restraints on the solute (includes the protein and ligand) followed by another round of free minimization. The systems were heated from 0 to 300\u00a0K on the protein and ligand in 100\u00a0ps. Then, the unrestrained systems were equilibrated for 1\u00a0ns with Langevin thermostat in the NPT (P\u00a0=\u00a01\u00a0atm and T\u00a0=\u00a0300\u00a0K) ensemble with Holm\u2013Sidak post hoc test and ANOVA test of linear mixed model for fix effect of main factors were performed. A two\u2010tailed P value of <\u00a00.05 was considered to be significant.Statistical analysis was performed with GraphPad Prism v7.03 . Normal distribution has been evaluated with the Shapiro\u2013Wilk test using GraphPad Prism. To determine statistical significance among investigated groups, Student\u2019s GP, AD, and SHEK conceived and designed the study, analyzed the data, and wrote the manuscript. GP designed and performed most of the experiments. JZ, JW, and H\u2010JM conducted microarray and data analysis. LH performed molecular docking. MD performed molecular dynamic simulation. PM\u2010A generated knockdown cells. PS participated in the mouse experiments. LL performed Fluidigm gene expression analysis. TAK provided cell lines, plasmids, and conceptual discussions. KE and CA generated HeLa inducible GFP, NOD1\u2010GFP, and NOD2\u2010GFP cells. HS performed Shigella infection experiments. NN, YD, MP, and IGB helped with the experiments. All the coauthors commented on the manuscript.The authors declare thet they have no conflict of interest.AppendixClick here for additional data file.Expanded View Figures PDFClick here for additional data file.Source Data for Expanded View and AppendixClick here for additional data file.Review Process FileClick here for additional data file.Source Data for Figure 4Click here for additional data file.Source Data for Figure 5Click here for additional data file."} +{"text": "In complex 1 the Re\u2014Re bond lengths [2.9767\u2005(3)\u20133.0133\u2005(2)\u2005\u00c5] are close to double the covalent Re radii (1.51\u2005\u00c5). The palladium\u2013rhenium carbonyl cluster 2 has not been structurally characterized previously; the Pd\u2014Re bond lengths [2.7582\u2005(2)\u20132.7796\u2005(2)\u2005\u00c5] are about 0.1\u2005\u00c5 shorter than the sum of the covalent Pd and Re radii (1.39\u00a0+\u00a01.51 = 2.90\u2005\u00c5). One carbene ligand and a diethyl ether mol\u00adecule are disordered over two positions with occupancy ratios of 0.5:0.5 and 0.625\u2005(15):0.375\u2005(15) in 1. An unidentified solvent is present in compound 2. The given chemical formula and other crystal data do not take into account the unknown solvent mol\u00adecule(s). The SQUEEZE routine [Spek and rhenium (Re) led to the isolation and crystallographic characterization of tetra\u00adkis\u00adpalladium(II) hexa\u00addeca\u00adcarbonyl\u00adtetra\u00adrhenium diethyl ether disolvate, [Pd(CSpek 2015. Acta Cr Two other products, tri\u00adphenyl\u00adphosphine oxide and the known complex Re2(CO)8(PPh3)2 and rhenium (Re) have important applications in alkane reforming, industrial chemical production, hydro\u00addechlorination and biomass conversion cation with four coordinated N-heterocyclic carbenes (NHC) lying on a twofold rotoinversion axis, and one [Re4(CO)16] anion. The geometry around the Pd atom is square-planar with one carbene unit being disordered. The C\u2014Pd\u2014C angles range from 86.9\u2005(4) to 97.7\u2005(4)\u00b0. The cluster anion lying on the inversion center has a perfectly flat rhombus geometry with the shortest Re\u2014Re bond [2.9767\u2005(3)\u2005\u00c5] corresponding to the short diagonal. The other four Re\u2014Re bond lengths [3.001\u2005(2)\u20133.0132\u2005(2)\u2005\u00c5] are also close to double the covalent Re radii \u20132.7796\u2005(2)\u2005\u00c5] are close to the sum of the covalent Pd and Re radii (1.39\u00a0+\u00a01.51 = 2.90\u2005\u00c5). In comparison, the Pd\u2014Re bond lengths in the PdRe4(CO)16(\u03bc-SbPh2)2(\u03bc-H)2 cluster \u201390.135\u2005(6)\u00b0 and the Pd\u2014Pd bond lengths are 2.9678\u2005(2)\u20132.99\u2005(2)\u2005\u00c5].The displacement ellipsoid plot of 1, each cation is surrounded by six anions and vice versa palladium(II) diiodide 16]2\u2212 anion, which is also found in the compound reported here. A search of the CSD found two closely related cluster compounds, viz. bis\u00ad(tetra\u00adethyl\u00adammo\u00adnium) hexa\u00addeca\u00adcarbonyl-tetra\u00adrhenium \u00adhexa\u00addeca\u00adcarbonyl\u00adtetra\u00adrhenium 4 was added to a toluene\u2013aceto\u00adnitrile mixture and 1,3-di\u00admethyl\u00adimidazolium-2-carboxyl\u00adate . The reaction mixture was refluxed for 1.5\u2005h, then Re2(CO)10 was added, the solution turned dark red and the solvents were removed in vacuo. The solid was washed with benzene (3 \u00d7 5\u2005ml) and recrystallized from an aceto\u00adnitrile\u2013di\u00adethyl\u00adether mixture. X-ray quality crystals of Pd(IMe)4Re4(CO)16\u00b72C4H10O were grown from a di\u00adchloro\u00admethane\u2013di\u00adethyl\u00adether mixture at 277\u2005K. 1HNMR : 3.41 , 7.37 . 13C{H} NMR : 36.9 , 123.5 , 168.0 , 197.7 (CO), 198.7 (CO), 201.1 (CO), 218.6 (CO) IR : 3152 , 1998 (vw), 1974 (vw), 1955 (m), 1927 (vw), 1912 (vw), 1881 , 1858 (vw), 1575 (vw), 1465 (w), 1400 (vw), 1332 (vw), 1229 (m), 1131 (vw), 1083 (vw), 1013 (vw), 845 (vw), 736 (s), 701 (vw), 681 (m), 600 (w), 577 (s), 560 (vw), 508 (vw), 496 (vw), 464 (w), 436 (vw), 411 (w).Under a nitro\u00adgen atmosphere, Pd(PPh4Re2(PPh3)4(m-CO)8(CO)2 suitable for X-ray diffraction analysis were obtained from a yellow benzene solution, after several days, by slow ether diffusion into a concentrated solution of benzene at 277\u2005K. IR : 3850 (vw), 3054 , 2955 , 1986 (s), 1821 , 1585 (vw), 1571 (vw), 1515 (vw), 1477 (w), 1434 (m), 1307 (vw), 1263 (vw), 1236 , 1182 (vw), 1159 (vw), 1119 (vw), 1092 (m), 1071 (vw), 1026 (vw), 997 (w), 907 (vw), 846 (vw), 741 (m), 690 (vs), 618 (vw), 565 (w), 541 (vw), 496 (m), 412 (vw).A few crystals of Pd2(CO)8(PPh3)2 were also isolated from this crystallization.Tri\u00adphenyl\u00adphosphine oxide and Resp2), 0.98\u2005\u00c5 (meth\u00adyl) and 0.99\u2005\u00c5 (methyl\u00adene), with common isotropic temperature factors for all hydrogen atoms of the aromatic rings and methyl groups. SADI restraints on bond lengths and DELU restraints on anisotropic thermal parameters were used to model the disordered carbene ligand and diethyl ether mol\u00adecule over two positions. For the refinement of 2, four reflections global, 1, 2. DOI: 10.1107/S2056989021009270/tx20411sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989021009270/tx20412sup3.hklStructure factors: contains datablock(s) 2. DOI: 2108168, 2108167CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Inter\u00admolecular N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds connect these dimers to form a three-dimensional network. In addition, C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions help to stabilize the packing.In the crystal, strong C\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds form dimers with 19H17N3O2, adopts a screw-boat conformation. In the crystal, strong C\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds form dimers with R22(14) and R22(12) ring motifs, respectively, between consecutive mol\u00adecules along the c-axis direction. Inter\u00admolecular N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds connect these dimers, forming a three-dimensional network. C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions contribute to the stabilization of the mol\u00adecular packing. A Hirshfeld surface analysis indicates that the contributions from the most prevalent inter\u00adactions are H\u22efH (47.1%), C\u22efH/H\u22efC (20.9%), O\u22efH/H\u22efO (15.3%) and N\u22efH/H\u22efN (11.4%).The central tetra\u00adhydro\u00adpyridine ring of the title compound, C The C7\u2013C12 phenyl ring, which is attached to N1, is in an equatorial position and makes a dihedral angle of 54.43\u2005(9)\u00b0 with the mean plane of the tetra\u00adhydro\u00adpyridine ring. The C13\u2013C18 meth\u00adoxy\u00adphenyl ring, which is attached to C4, is in an axial position. The dihedral angle between the C7\u2013C12 phenyl and C13\u2013C18 meth\u00adoxy\u00adphenyl rings is 68.61\u2005(10)\u00b0.The title compound Fig.\u00a01 crystallC\u22efN2 hydrogen bonds \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01; (v) \u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a0Cg3 is the centroid of the C13\u2013C18 meth\u00adoxy\u00adphenyl ring; Table\u00a01Cg2\u22efCg2iii = 3.8918\u2005(15)\u2005\u00c5 and slippage = 1.551\u2005\u00c5; Cg2 is the centroid of the C7\u2013C12 phenyl ring] contribute to the stabilization of the mol\u00adecular packing to 1.8469 (blue) a.u. The dnorm mapping indicates that strong hydrogen-bonding inter\u00adactions, such as N\u2014H\u22efN, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds , followed by C\u22efH/H\u22efC , O\u22efH/H\u22efO and N\u22efH/H\u22efN . The percentage contributions of the C\u22efC, C\u22efN/N\u22efC and N\u22efN contacts are negligible, at 3.1, 1.4 and 0.8%, respectively. The predominance of H\u22efH, C\u22efH/H\u22efC, O\u22efH/H\u22efO and N\u22efH/H\u22efN contacts indicate that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing -6-oxo-1-phenyl-1,4,5,6-tetra\u00adhydro\u00adpyridine-3-carbo\u00adnitrile (I) -2-(pyridin-2-yl)ethen\u00adyl]-1,4,5,6-tetra\u00adhydro\u00adpyridine-3-carboxyl\u00adate (III) crystallizes in the monoclinic space group Pc with Z = 4, and with two mol\u00adecules, A and B, in the asymmetric unit. These mol\u00adecules are stereoisomers with an R,R absolute configuration at C3 and C4 in mol\u00adecule A, whereas the corresponding atoms in B, C23 and C24, have an S configuration. In both mol\u00adecules, the conformation of the central di\u00adhydro\u00adpyridine ring is close to screw-boat. The mol\u00adecular conformation is stabilized by N\u2014H\u22efO hydrogen bonds, forming a dimer with an c-axis direction. Furthermore C\u2014Br\u22ef\u03c0 and C=O\u22ef\u03c0 stacking inter\u00adactions between these ribbons contribute to the stabilization of the mol\u00adecular packing.Compound (II) crystallizes in the monoclinic space group P21/c with Z = 4 and the asymmetric unit comprises one mol\u00adecule. The central tetra\u00adhydro\u00adpyridine ring is almost planar with a maximum deviation of 0.074\u2005(3)\u2005\u00c5 for C4. The phenyl and di\u00adchloro\u00adphenyl rings are at an angle of 21.28\u2005(15)\u00b0. They form dihedral angles of 86.10\u2005(15) and 87.17\u2005(14)\u00b0, respectively, with the central tetra\u00adhydro\u00adpyridine ring. A strong intra\u00admolecular O2\u2014H2\u22efO1 hydrogen bond stabilizes the mol\u00adecular conformation of the mol\u00adecule, creating an S(6) ring motif. In the crystal, mol\u00adecules are linked by inter\u00admolecular N\u2014H\u22efN and C\u2014H\u22efN hydrogen bonds, and N\u2014H\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional network.Compound (III) , the cis configuration of the pyridinyl-vinyl fragment is stabilized by a strong intra\u00admolecular N\u2014H\u22efN hydrogen bond. The phenyl and pyridine rings are inclined to one another by 77.3\u2005(1)\u00b0. In the crystal, inversion dimers are present via pairs of C\u2014H\u22efO hydrogen bonds and are further linked by C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions.In mol\u00adecule (IV) , the mol\u00adecules form dimers by means of a pair of N\u2014H\u22efO hydrogen bonds. The 2(1H)-pyridone ring displays a screw-boat conformation.For compound (To a solution of 2-(4-meth\u00adoxy\u00adbenzyl\u00adidene)malono\u00adnitrile and acetoacetanilide in methanol (25\u2005mL), 3-4 drops of piperidine were added and the mixture was stirred at 328\u2013333\u2005K for 10\u2005min and was kept at room temperature for 48\u2005h. Then 15\u2005mL of methanol were removed from the reaction mixture, which was left overnight. The precipitated crystals were separated by filtration and recrystallized from ethanol/water (1:1) solution .1H NMR : 2.80 ; 3.19 ; 3.82 ; 3.93 ; 5.85 ; 7.15\u20137.58 . 13C NMR : 36.06 (CH\u2014Ar), 40.42 (CH2), 53.78 (OCH3), 59.05 (Cquat.), 112.89 (2CHar), 121.21 (CN), 128.61 (CHar.), 128.88 (2CHar.), 130.44 (2CHar.), 130.51 (2CHar.), 136.06 (Car. quat.), 137.02 (Car. quat.), 154.59 (Car. quat.), 155.18 (Cquat.), 168.82 (N\u2014C=O).C = 0.91\u2005(2) and N3\u2014H3D = 0.91\u2005(2)\u2005\u00c5 with Uiso(H) = 1.2Ueq(N)]. C-bound H atoms were positioned geometrically, with C\u2014H = 0.95\u20131.00\u2005\u00c5, and were refined with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698902200175X/vm2260sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902200175X/vm2260Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698902200175X/vm2260Isup3.cmlSupporting information file. DOI: 2152191CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "This compound exists in the crystal phase as a methanol solvate.The mol\u00adecular and crystal structures of the title compound, C 36H37ClN4O7\u00b7CH3OH, which crystallizes as a methanol solvate, may possess biological activity, which is inherent for a natural peptide or protein. In the crystal, mol\u00adecules of the title compound form hydrogen-bonded tetra\u00admers with the solvate mol\u00adecules acting as bridges as a result of the O\u2014H\u22efO and N\u2014H\u22efO inter\u00admolecular hydrogen bonds. Hirshfeld surface analysis was used to study the different types of inter\u00admolecular inter\u00adactions whose contributions are: H\u22efH = 53.8%, O\u22efH/H\u22efO = 19.0%, C\u22efH/H\u22efC = 14.8%, Cl\u22efH/H\u22efCl = 5.3%, N\u22efH/H\u22efN = 3.2%.The title compound, C The target product contains a heterocyclic core bound to peptidomimetics, compounds that mimic a natural peptide or protein and which may have high biological activity. The pyrrolone fragment is also a privileged motif because of its biological activities, namely anti\u00adbacterial with the populations of A:B in a 0.303\u2005(10):0.697\u2005(10) ratio. All atoms of the partially saturated five-membered heterocycle are in the same plane with an accuracy of 0.008\u2005\u00c5. The N2\u2014C4 bond length of 1.380\u2005(3)\u2005\u00c5 and the C8\u2014N2\u2014C4\u2014N1 torsion angle of 2.2\u2005(4)\u00b0 indicate conjugation between the \u03c0-systems of the partially saturated and oxazole cycles. The para-chloro\u00adphenyl substituent is located in the pseudo-equatorial position and is turned in relation to the C7\u2014C8 endocyclic bond [C6\u2014C7\u2014C8\u2014C9 = \u2212120.9\u2005(3)\u00b0 and C7\u2014C8\u2014C9\u2014C10 = 60.9\u2005(3)\u00b0]. The C16(=O4)\u2014N3 carb\u00adamide group is located in the -ac position in relation to the C6\u2014C7 endocyclic bond [C6\u2014C7\u2014C15\u2014C16 = \u2212107.2\u2005(3)\u00b0], and the C16=O4 carbonyl group is slightly non-coplanar to the C7\u2014C15 bond [C7\u2014C15\u2014C16\u2014O4 = 22.5\u2005(4)\u00b0]. The para-meth\u00adoxy\u00adphenyl substituent at the nitro\u00adgen atom is turned almost orthogonally to the plane of the carbamide group [C16\u2014N3\u2014C17\u2014C22 = \u221299.5\u2005(3)\u00b0]. The para-meth\u00adoxy\u00adphenyl substituent at the carbon atom is located in a position inter\u00admediate between sp and \u2212sc and is also rotated almost orthogonally to the plane of the carbamide group . In both para-meth\u00adoxy\u00adphenyl substituents, the meth\u00adoxy group is coplanar with the plane of the aromatic ring despite the steric repulsion between the methyl group and the aromatic ring atoms . The substituent at the C24 atom is located in the \u2212sc position relative to the N3\u2014C16 bond [C32\u2014C24\u2014N3\u2014C16 = \u221278.1\u2005(3)\u00b0] and the C32\u2014O6 carbonyl group is slightly non-coplanar to the N3\u2014C24 bond [O6\u2014C32\u2014C24\u2014N3 = \u221227.8\u2005(3)\u00b0]. The tert-butyl substituent is located in anti\u00adperplanar position to the C32\u2014C24 bond [C33\u2014N4\u2014C32\u2014C24 = 172.9\u2005(2)\u00b0].The title compound crystallizes as a methanol solvate Fig.\u00a01. The metA\u22efO8A, O3\u2014H8B\u22efO8B, O8A\u2014H8A\u22efO4 and O8B\u2013H8B\u22efO4 inter\u00admolecular hydrogen bonds . The highest contribution is from H\u22efH contacts (53.8%), while these made by the O\u22efH/H\u22efO (19.0%) and C\u22efH/H\u22efC (14.8%) inter\u00adactions are similar . The contributions of Cl\u22efH/H\u22efCl (5.3%) and N\u22efH/H\u22efN (3.2%) inter\u00adactions are very small.In the two-dimensional fingerprint plots the pair of sharp spikes indicate strong hydrogen bonds and short contacts in the crystal structure Fig.\u00a04a. The har Fig.\u00a04c, 4d. Tns Fig.\u00a04e, 4f aret al., 2016et al., 2019et al., 2018et al., 2003bet al., 2011et al., 2013et al., 2017et al., 2003aet al., 2011et al., 2009et al., 2015et al., 2005et al., 2019et al., 2019et al., 2008et al., 2016et al., 2018et al., 1991et al., 2006et al., 1990et al., 2016et al., 2001et al., 2019et al., 2004et al., 2014A search of the Cambridge Structural Database (0.5\u2005mmol) and 4-meth\u00adoxy\u00adbenzaldehyde (2) (0.5\u2005mmol) were dissolved in methanol (4\u2005mL) and stirred for 0.5\u2005h. Then, 2-acetic acid (1) (0.5\u2005mmol) and tert-butyl\u00adisocyanide (4) (0.5\u2005mmol) were added consistently and the reaction mixture was stirred for 24\u2005h at 319\u2005K. The mixture was allowed to stand overnight. The crystal precipitate was filtered off and dried. The reaction scheme is shown in Fig.\u00a054-Meth\u00adoxy\u00adaniline (Uiso(H) = 1.5Ueq for methyl and hydroxyl groups and with Car\u2014H = 0.93\u2005\u00c5, Csp3\u2014H = 0.97\u2005\u00c5, N\u2014H = 0.89\u2005\u00c5 Uiso(H) = 1.2Ueq for all other hydrogen atoms. The solvent molecule is disordered over two positions (A and B) in a 0.303\u2005(10):0.697\u2005(10) occupancy ratio. Csp3\u2014O bonds were refined with fixed length of 1.413\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989021011312/zq2268sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021011312/zq2268Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021011312/zq2268Isup3.cmlSupporting information file. DOI: 2118094CCDC reference: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compounds feature O\u2014H\u22efO carb\u00adoxy\u00adlic-acid inversion dimers and N\u2014H\u22efN chains in their extended structures. 11H11N3O3, (I), and C10H10N2O2, (II), are commercially available and were crystallized from ethyl acetate solution. The dihedral angle between the pyrazole and phenyl rings in (I) is 52.34\u2005(7)\u00b0 and the equivalent angle between the isoxazole and phenyl rings in (II) is 7.30\u2005(13)\u00b0. In the crystal of (I), the mol\u00adecules form carb\u00adoxy\u00adlic acid inversion dimers with an R(8) ring motif via pairwise O\u2014H\u22efO hydrogen bonds. In the crystal of (II), the mol\u00adecules are linked via N\u2014H\u22efN hydrogen bonds forming chains propagating along [010] with a C(5) motif. A weak N\u2014H\u22ef\u03c0 inter\u00adaction also features in the packing of (II). Hirshfeld surface analysis was used to explore the inter\u00admolecular contacts in the crystals of both title compounds: the most important contacts for (I) are H\u22efH (41.5%) and O\u22efH/H\u22efO (22.4%). For (II), the most significant contact percentages are H\u22efH (36.1%) followed by C\u22efH/H\u22efC (31.3%).The title compounds, C The C3\u2014N3 distance of 1.353\u2005(2)\u2005\u00c5 is typical for an amino group bound to an aromatic ring. The carb\u00adoxy\u00adlic carbon\u2013oxygen distances are 1.255\u2005(2) and 1.316\u2005(2) for C4\u2014O2 and C4\u2014O1, respectively, indicating that the former bond may be affected by the intra\u00admolecular N\u2014H\u22efO hydrogen bond.The mol\u00adecular structure of compound (I)d Table\u00a01. This boif Fig.\u00a01 with an Database survey section). The C3\u2014N2 distance is 1.350\u2005(5)\u2005\u00c5 and is typical of an amino group bound to an aromatic ring.The mol\u00adecular structure of compound (II)via the O1\u2014H1\u22efO2i link to generate an R(8) loop with O\u22efO = 2.649\u2005(2)\u2005\u00c5, see Table\u00a01via \u03c0\u2013\u03c0 inter\u00adactions, notably weak stacking inter\u00adactions between the 4-meth\u00adoxy\u00adphenyl rings along the a-axis direction.In the extended structure of (I)b-axis direction via the N2\u2014H2A\u22efN1i hydrogen bond , C6\u2014H6\u22efCg1iii (2.86\u2005\u00c5) and C9\u2014H9\u22efCg2ii (2.86\u2005\u00c5) . Analysis of the two-dimensional fingerprint plots indicates that the H\u22efH (36.1%) inter\u00adactions are the major factor in the crystal packing with C\u22efH/H\u22efC (31.3%) contacts making the next highest contribution. The percentage contributions of other weak inter\u00adactions are: O\u22efH/H\u22efO (17.3%) and N\u22efH/ H\u22efN (12.1%). Figures showing the shape-index surface for each compound and the overall fingerprint plots are included in the supporting information.Fig.\u00a06et al., 2016et al., 1974et al., 2007N-4-dimethyl-N-[3-{4-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl]-1,2-oxazol-5-yl}benzene-1-sulfonamide -1,2-oxazole -5-nitro-4,5-di\u00adhydro-1H-pyrazole-4-carboxyl\u00adate Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C). In order to ensure a chemically meaningful O\u2014H distance in (I)Uiso(H) = 1.5Ueq(O). In (I)Uiso(H) = 1.5Ueq(N). The absolute structure of (II)Crystal data, data collection, and structure refinement details are summarized in Table\u00a0310.1107/S2056989022001827/hb8013sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989022001827/hb8013Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989022001827/hb8013IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989022001827/hb8013Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022001827/hb8013IIsup5.cmlSupporting information file. DOI: 10.1107/S2056989022001827/hb8013sup6.pdfShape-index and fingerprint plots for compounds (I) and (II). DOI: 2152583, 2152582CCDC references: crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Neighbouring (C 12H15N2)2[ZnCl4], is a salt with two symmetrically independent, essentially planar heterocyclic cations and a slightly distorted tetra\u00adhedral chloro\u00adzincate dianion. N\u2014H\u22efCl hydrogen bonds link these ionic constituents into a discrete aggregate, which comprises one formula unit. The effect of hydrogen bonding is reflected in the minor distortions of the [ZnCl4]2\u2212 moiety: distances between the cation and chlorido ligands engaged in classical hydrogen bonds are significantly longer than the others. Secondary inter\u00adactions comprise C\u2014H\u22ef\u03c0 hydrogen bonding and weak \u03c0\u2013\u03c0 stacking. A Hirshfeld surface analysis indicates that the most abundant contacts in packing stem from H\u22efH (47.8%) and Cl\u22efH/H\u22efCl (29.3%) inter\u00adactions.The title compound, (C Mackinlaya sp. tetra\u00adchlorido\u00adzincate, an inter\u00admediate in the synthesis of mackinazolinone, using high-resolution diffraction data and Hirshfeld surface analysis.Tricyclic quinazolines are counted among the most exciting quinazoline alkaloids. Specifically, the alkaloid mackinazoline was isolated from P21/n space group, with two [C12H15N2]+ cations and a [ZnCl4]2\u2212 counter-anion in the asymmetric unit are essentially coplanar, with r.m.s. deviations of 0.0437 and 0.0168\u2005\u00c5 for mol\u00adecules A and B, respectively. The remaining atoms of the third ring are significantly displaced above the opposite faces of these planes with deviations of 0.3877\u2005(12)\u2005\u00c5 for C10A and 0.3831\u2005(11)\u2005\u00c5 for C11A in residue A and 0.4705\u2005(11)\u2005\u00c5 for C10B and 0.2495\u2005(11)\u2005\u00c5 for C11B in residue B. Fig.\u00a02The title compound crystallizes in the it Fig.\u00a01. The benA and N1B, respectively, and the acquired positive charge is delocalized within the \u2013N\u2013C\u2013N\u2013 moiety in the ring, where the N1A\u2014C2A and N1B\u2014C2B bonds are only slightly longer than C2A\u2014N3A and C2B\u2014N3B fragment (see Table S1 in the supporting information).However, these C\u2014N bond lengths are shorter than those in the related tricyclic protonated and the very favourable ratio between observations and variables (100:1) in our diffraction data result in small standard uncertainties for atomic coordinates and derived geometric parameters and allow to discuss more subtle details. The most acute angle of 103.33\u2005(11)\u00b0 within the tetra\u00adchlorido\u00adzincate dianion graph-set motif \u2005\u00c5] and involves an inter\u00adaction between the electron-rich equatorial region of the halogen atom and the ring atom attached to two N-atom neighbours, most probably the most electron-deficient atom in the heterocycle. These contacts link anions and cations into a three-dimensional network. Weak \u03c0\u2013\u03c0 stacking inter\u00adactions occur between pyrimidine and benzene rings of anti\u00adparallel pairs of cations and involve contact distances of Cg1\u22efCg3 = 3.6225\u2005(5)\u2005\u00c5 (slippage 0.857\u2005\u00c5) and of Cg7\u22efCg9 = 3.6246\u2005(7)\u2005\u00c5 (slippage 0.994\u2005\u00c5).The crystal packing is further stabilized by inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions Table\u00a02 and addiCrystalExplorer17.5 analysis in Fig.\u00a06ly Fig.\u00a06. Minor cet al., 20164]2\u2212 anions. They are associated with refcodes PODLUP of 2,3-tetra\u00admethyl\u00adenquinazoline-4-one Fig.\u00a07 were plaUiso(H) = 1.2Ueq(C). H atoms bonded to nitro\u00adgen were located in a difference-Fourier map, and their positional and isotropic displacement parameters were freely refined.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989021004989/jq2006sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021004989/jq2006Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021004989/jq2006sup4.docxSupporting information file. DOI: 2082994CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Both polymorphs are monoclinic and the [Rh(\u03bc-I)(COD)]2 mol\u00adecules in each case possess C2v symmetry. However, the core geometry of the butterfly-shaped Rh2I2 core differs substanti\u00adally. In the C2/c polymorph, the core geometry of [Rh(\u03bc-I)(COD)]2B is bent, with a hinge angle of 96.13\u2005(8)\u00b0 and a Rh\u22efRh distance of 2.9612\u2005(11)\u2005\u00c5. The P21/c polymorph features a more planar [Rh(\u03bc-I)(COD)]2P core geometry, with a hinge angle of 145.69\u2005(9)\u00b0 and a Rh\u22efRh distance of 3.7646\u2005(5)\u2005\u00c5.The solid-state structure of di-\u03bc-iodido-bis\u00ad{rhodium(I)}, [Rh M(\u03bc-X)(COD)]2, are ubiquitous synthons for Rh and Ir catalysts. As a representative example, [Rh(COD)(DPEphos)]BF4 catalyzes the hydro\u00adamination of vinyl\u00adarenes with anti-Markovnikov selectivity, and is prepared via the reaction of [Rh(\u03bc-Cl)(COD)]2 with two equivalents of AgBF4 and two equivalents of DPEphos (COD)]2 , all compounds have been structurally characterized with the notable exception of [Rh(\u03bc-I)(COD)]2 (COD)]2 containing dimers with significantly different structural features. X-ray diffraction of the dark-orange crystals revealed that [Rh(\u03bc-I)(COD)]2 crystallized in the monoclinic C2/c space group in this polymorph. The mol\u00adecular structure of [Rh(\u03bc-I)(COD)]2B exhibited a C2v-symmetric geometry featuring a bent Rh2I2 diamond core (COD)]2 and [Rh(\u03bc-Br)(COD)]2, which both exhibit more planar Rh2X2 cores with hinge angles of 169.08\u2005(6) and 148.74\u2005(7)\u00b0 and Rh\u22efRh distances of 3.5169\u2005(6) and 3.5648\u2005(14)\u2005\u00c5, respectively (COD)]2 re Fig.\u00a01. The hin2, this time crystallizing in the monoclinic P21/c space group. The geometry of the dirhodium dimer in the P21/c polymorph [Rh(\u03bc-I)(COD)]2P differs significantly from the C2/c polymorph [Rh(\u03bc-I)(COD)]2B. The Rh2I2 core geometry of [Rh(\u03bc-I)(COD)]2P is more planar, with a hinge angle of 145.69\u2005(9)\u00b0 and a Rh\u22efRh distance of 3.7646\u2005(5)\u2005\u00c5 (COD)]2 and [Rh(\u03bc-Br)(COD)]2 (COD)]\u2005\u00c5 Fig.\u00a02. The mol\u22121) between planar and bent [Rh(\u03bc-X)(L)2]2 geometries (COD)]2B but not [Rh(\u03bc-Cl)(COD)]2 and [Rh(\u03bc-Br)(COD)]2 (COD)]2.A previous theoretical study found relatively small energetic differences and 2.6975\u2005(7)\u2005\u00c5]. The four Rh\u2014I distances in [Rh(\u03bc-I)(COD)]2P are slightly less symmetric: the bonds between I2 and the two Rh centers [2.6833\u2005(4) and 2.6738\u2005(4)\u2005\u00c5] are slightly shorter than those associated with I1 [2.6998\u2005(4) and 2.7061\u2005(4)\u2005\u00c5]. Similarly, the Rh\u2014C distances in [Rh(\u03bc-I)(COD)]2B are more symmetric, ranging from 2.115\u2005(6) to 2.122\u2005(6)\u2005\u00c5, while the Rh\u2014C distances in [Rh(\u03bc-I)(COD)]2P range from 2.117\u2005(4) to 2.131\u2005(4)\u2005\u00c5. The average Rh\u2014C distance in the bent and planar structures are similar to the average Rh\u2014C distances reported for the [Rh(\u03bc-Cl)(COD)]2 and [Rh(\u03bc-Br)(COD)]2 analogues (COD)]2P are considerably longer than the average Rh\u2014Br and Rh\u2014Cl distances in [Rh(\u03bc-Br)(COD)]2 and [Rh(\u03bc-Cl)(COD)]2 .There is no meaningful difference between the two independent Rh\u2014I distances in [Rh(\u03bc-I)(COD)]2 are attributed to differences in crystal packing and weak inter\u00adatomic forces. The bent and planar geometries are likely similar in energy. Stabilization of the bent geometry in [Rh(\u03bc-I)(COD)]2B arises from intra\u00admolecular dispersion forces between the C\u2014H bonds of the cyclo\u00adocta\u00addiene ligands on the two Rh centers within each mol\u00adecule. Indeed there are four close C\u2014H\u22efH\u2014C contacts between the alkene and methyl\u00adene hydrogen atoms made possible by the bent geometry (COD)]2P are two Rh\u22efH\u2014C contacts in the apical positions of Rh2 = 2.67\u2005\u00c5; Rh2\u22efH3A = 2.93\u2005\u00c5) that could, at best, be labeled as weak inter\u00admolecular agostic inter\u00adactions than the bent C2/c polymorph (2.597\u2005g\u2005cm\u22123), indicative of tighter crystal packing.The structural differences between the dimers in the two polymorphs of [Rh(\u03bc-I)(COD)]ry Fig.\u00a03. In the ns Fig.\u00a04. Consist2 was prepared according to the procedure described by J. A. Hlina et al. was added to toluene (5\u2005mL) and tri\u00admethyl\u00adsilyl iodide . The reaction mixture turned dark red and rust-colored crystals precipitated from the solution. The solid was isolated, washed with hexa\u00adnes and dried in vacuo to yield the final product [Rh(\u03bc-I)(COD)]2 as a red\u2013brown crystalline solid . X-ray quality crystals were grown from a concentrated solution of toluene at 236\u2005K, resulting in crystals with two different morphologies. 1H NMR (C6D6): 1.15\u20131.28 , 1.90\u20132.05 , 4.62\u20134.70 . A single species was observed by 1H NMR spectroscopy and the 1H NMR spectrum did not contain any broad features indicative of dynamic behavior or inter\u00adconversion between the two isomers in solution.[Rh(\u03bc-I)(COD)] al. 2017. Under aSHELXT) produced a complete phasing model for refinement. All non-hydrogen atoms were refined anisotropically by full-matrix least-squares (SHELXL2018). All carbon-bonded methyl\u00adene hydrogen atoms were placed using a riding model. Their positions were constrained relative to their parent atom using the appropriate HFIX command in SHELXL2018. All carbon-bonded methine hydrogen atoms were located in the difference map. Their C\u2014H distances were restrained to a target value of 1.00\u2005(2)\u2005\u00c5. For all H atoms, displacement parameter Uiso(H) values were set to 1.2 times Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S205698902100743X/zl5015sup1.cifCrystal structure: contains datablock(s) global, B, P. DOI: 10.1107/S205698902100743X/zl5015Bsup2.hklStructure factors: contains datablock(s) B. DOI: 10.1107/S205698902100743X/zl5015Psup3.hklStructure factors: contains datablock(s) P. DOI: 2097568, 2097567CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Saccharomyces cerevisiae is its ability to perform simultaneous respiration and fermentation at high growth rate even under fully aerobic conditions. In the present work, this Crabtree effect called phenomenon was investigated in detail by comparative 13C metabolic flux analysis of S. cerevisiae growing under purely oxidative, respiro-fermentative and predominantly fermentative conditions.One of the most fascinating properties of the biotechnologically important organism S. cerevisiae exhibited a by-pass of pyruvate dehydrogenase in all physiological regimes. During oxidative growth this by-pass was mainly provided via pyruvate decarboxylase, acetaldehyde dehydrogenase, acetyl-CoA synthase and transport of acetyl-CoA into the mitochondrion. During fermentative growth this route, however, was saturated due to limited enzyme capacity. Under these conditions the cells exhibited high carbon flux through a chain of reactions involving pyruvate carboxylase, the oxaloacetate transporter and malic enzyme. During purely oxidative growth the PPP alone was sufficient to completely supply NADPH for anabolism. During fermentation, it provided only 60 % of the required NADPH.The metabolic shift from oxidative to fermentative growth was accompanied by complex changes of carbon flux throughout the whole central metabolism. This involved a flux redirection from the pentose phosphate pathway (PPP) towards glycolysis, an increased flux through pyruvate carboxylase, the fermentative pathways and malic enzyme, a flux decrease through the TCA cycle, and a partial relocation of alanine biosynthesis from the mitochondrion to the cytosol. S. cerevisiae possesses different metabolic by-passes to channel carbon into the mitochondrion. This involves the conversion of cytosolic pyruvate either into acetyl CoA or oxaloacetate followed by intercompartmental transport of these metabolites. During oxidative growth mainly the NAD specific isoforms of acetaldehyde dehydrogenase and isocitrate dehydrogenase catalyze the corresponding reactions in S. cerevisiae, whereas NADPH supply under fermentative conditions involves significant contribution of sources other than the PPP such as e. g. NADPH specific acetaldehyde dehydrogenase or isocitrate dehydrogenase.We conclude that, in order to overcome the limited capacity of pyruvate dehydrogenase, S. cerevisiae is an important biotechnological organism e.g. for production of ethanol [S. cerevisiae is able to perform simultaneous respiration and fermentation at high growth rates even under fully aerobic conditions [S. cerevisiae by a shift from biomass formation towards fermentative products at increased dilution rates [13C metabolic flux analysis which allows the quantification of in vivo activity of pathways and reactions [S. cerevisiae and related yeast strains [S. cerevisiae by quantifying metabolic fluxes through the underlying reactions in the central metabolism at these different physiological states. For this purpose chemostat cultures with [1-13C] glucose as substrate were grown at different growth rates under aerobic glucose-limited conditions to metabolic and isotopic steady-state. The use of a continuous culture hereby allowed the precise adjustment of the physiological state of the cells, i.e. the relative activity of the fermentative pathways.The budding yeast ethanol , recombi ethanol , antibio ethanol or bioma ethanol . Additio ethanol ,6. This nditions ,8. This on rates . Despiteon rates -12, a clon rates ,14. The on rates ,16. A poeactions -19. In r strains -27. In t-1 and 0.45 h-1 , mixed respiro-fermentative (0.30 h-1), and mainly fermentative metabolism (0.40 h-1) were selected for the flux studies. The resulting ethanol yield under these conditions was 0.0, 0.5, and 1.1 mol ethanol (mol glucose)-1, respectively. The stoichiometric data obtained from the 13C cultures agreed well with the corresponding values from the initially performed cultivations The metabolic shift had a strong effect on the flux partitioning between glycolysis and PPP. Whereas, at purely oxidative metabolism, the major carbon flux was channeled into the PPP, the opposite was observed for fermentative metabolism. The data of the present work together with previous flux studies of t Figure . A simil 0.4 h-1 . The red 0.4 h-1 . Due to S. cerevisiae glucose . Cultivations were performed in a continuously operated bioreactor with a culture volume of 100 ml at 30\u00b0C and 500 rpm. The aeration rate was kept at 100 mL/min by a mass flow controller . Composition of inlet and exhaust gas was determined on-line by mass spectrometry as described previously was obtained. Enzymatic test-kits were used for quantification of glucose, glycerol, acetate and ethanol in the supernatant. Concentrations of 2-oxoglutarate, pyruvate, succinate, and fumarate in the supernatant were analyzed by HPLC [S. cerevisiae was quantified by HPLC [i.e. the quantification of the yield coefficients, was 3.7 % (biomass yield), 4.0 % (ethanol yield), 9.3 % (acetate yield) and 11.3 % (glycerol yield).Optical density (OD by HPLC after hy0) to the fully 13C labeled (xn) variant could be measured with good signal intensity and accuracy is shown in Figure S. cerevisiae [The metabolic network of revisiae . For glyrevisiae ,46 was t1 - \u03bd2 - \u03bd3 + \u03bd4 - \u03bd5 = 0 \u00a0\u00a0\u00a0 (1)Glucose 6-phosphate: \u03bd3 - \u03bd4 - \u03bd6 + \u03bd8 - \u03bd9 + \u03bd10 - \u03bd11 = 0 \u00a0\u00a0\u00a0 (2)Fructose 6-phosphate: \u03bd2 - \u03bd7 - \u03bd8 + \u03bd9 - 2\u03bd12 + 2\u03bd13 = 0 \u00a0\u00a0\u00a0 (3)Pentose phosphate: \u03bd8 + \u03bd9 + \u03bd10 - \u03bd11 - \u03bd14 = 0 \u00a0\u00a0\u00a0 (4)Erythrose 4-phosphate: - \u03bd12 - \u03bd13 - \u03bd10 + \u03bd11 - \u03bd5 = 0 \u00a0\u00a0\u00a0 (5)Sedoheptulose 7-phosphate: \u03bd6 - \u03bd10 + \u03bd11 + \u03bd8 - \u03bd9 + \u03bd12 - \u03bd13 + \u03bd16 - \u03bd17 - \u03bd18 = 0 \u00a0\u00a0\u00a0 (6)Glyceraldehyde 3-phosphate: \u03bd6 - \u03bd15 - \u03bd16 = 0 \u00a0\u00a0\u00a0 (7)Dihydroxyacetone-phosphate: \u03bd17 - \u03bd19 - \u03bd20 - \u03bd24 = 0 \u00a0\u00a0\u00a0 (8)3-Phosphoglycerate: \u03bd20 - \u03bd21 - \u03bd22 = 0 \u00a0\u00a0\u00a0 (9)Serine: \u03bd22 - \u03bd23 + \u03bd29 = 0 \u00a0\u00a0\u00a0 (10)Glycine: \u03bd28 - \u03bd29 - \u03bd30 = 0 \u00a0\u00a0\u00a0 (11)Threonine: \u03bd24 - \u03bd25 - \u03bd26 = 0 \u00a0\u00a0\u00a0 (12)Phosphoenolpyruvate: \u03bdcyt: \u03bd26 - \u03bd27 - \u03bd32 - \u03bd33 - \u03bd34 = 0 \u00a0\u00a0\u00a0 (13)Pyruvate32 - \u03bd37 - \u03bd38 = 0 \u00a0\u00a0\u00a0 (14)Acetaldehyde: \u03bd38 - \u03bd39 - \u03bd40 = 0 \u00a0\u00a0\u00a0 (15)Acetate: \u03bdcyt: \u03bd40 - \u03bd41 - \u03bd47 + \u03bd48 = 0 \u00a0\u00a0\u00a0 (16)Acetyl CoAcyt: \u03bd33 - \u03bd42 = 0 \u00a0\u00a0\u00a0 (17)Alaninecyt: \u03bd27 - \u03bd28 - \u03bd31 - \u03bd35 + \u03bd36 = 0 \u00a0\u00a0\u00a0 (18)Oxaloacetatemit: \u03bd34 - \u03bd43 - \u03bd44 + \u03bd50 - \u03bd51 = 0 \u00a0\u00a0\u00a0 (19)Pyruvatemit: \u03bd43 - \u03bd45 = 0 \u00a0\u00a0\u00a0 (20)Alaninemit: \u03bd44 - \u03bd46 + \u03bd47 - \u03bd48 - \u03bd49 = 0 \u00a0\u00a0\u00a0 (21)Acetyl CoA49 - \u03bd52 - \u03bd53 = 0 \u00a0\u00a0\u00a0 (22)2-Oxoglutarate: \u03bd53 - \u03bd54 + \u03bd55 = 0 \u00a0\u00a0\u00a0 (23)Succinate: \u03bdmit: \u03bd35 - \u03bd36 - \u03bd49 - \u03bd50 + \u03bd54 - \u03bd55 = 0 \u00a0\u00a0\u00a0 (24)Oxaloacetate/MalateS. cerevisiae was considered combining metabolite balancing and isotopomer modeling. The performed approach utilized metabolite balancing during each optimization step considering stoichiometric data on anabolic demand for biomass precursors Table and prod13C tracer studies and all analytics involved.OF carried out the experimental work including the chemostat cultivations, the S. cerevisiae, the software implementation, the calculation of carbon fluxes, and the statistical analysis. He drafted and composed the manuscript.CW designed the study and carried out all computational work involving the construction of the metabolic network model of Both authors read and approved the final manuscript."} +{"text": "CENP\u2010A nucleosomes. Centromere maintenance during the cell cycle requires HJURP\u2010mediated CENP\u2010A deposition, a process regulated by the Mis18 complex (Mis18\u03b1/Mis18\u03b2/Mis18BP1). Spatial and temporal regulation of Mis18 complex assembly is crucial for its centromere association and function. Here, we provide the molecular basis for the assembly and regulation of the Mis18 complex. We show that the N\u2010terminal region of Mis18BP1 spanning amino acid residues 20\u2013130 directly interacts with Mis18\u03b1/\u03b2 to form the Mis18 complex. Within Mis18\u03b1/\u03b2, the Mis18\u03b1 MeDiY domain can directly interact with Mis18BP1. Mis18\u03b1/\u03b2 forms a hetero\u2010hexamer with 4 Mis18\u03b1 and 2 Mis18\u03b2. However, only two copies of Mis18BP1 interact with Mis18\u03b1/\u03b2 to form a hetero\u2010octameric assembly, highlighting the role of Mis18 oligomerization in limiting the number of Mis18BP1 within the Mis18 complex. Furthermore, we demonstrate the involvement of consensus Cdk1 phosphorylation sites on Mis18 complex assembly and thus provide a rationale for cell cycle\u2010regulated timing of Mis18 assembly and CENP\u2010A deposition.The centromere, a chromosomal locus that acts as a microtubule attachment site, is epigenetically specified by the enrichment of Equal and identical distribution of chromosomes to each daughter cell during cell division is essential for maintaining genome integrity. A central player regulating this process is the kinetochore, a large proteinaceous structure assembled at a specialized region of the chromosome called the centromere Schizosaccharomyces pombe); however, the molecular mechanisms by which it is achieved share considerable similarity In most eukaryotes, the centromeric chromatin is epigenetically defined by the enrichment of nucleosomes containing the histone H3 variant CENP\u2010A S. pombe and humans The Mis18 proteins contain two structurally distinct domains , both of which can self\u2010oligomerize. Previously, we and others have shown that oligomerization of Mis18 proteins is required for their centromere association and function both in 20\u2013130), binds Mis18\u03b1/\u03b2 hetero\u2010oligomer via the Mis18\u03b1/\u03b2 MeDiY hetero\u2010dimer to form the Mis18 complex. Characterization of the oligomeric structures of the Mis18 subcomplexes revealed that the Mis18\u03b1/\u03b2 complex is a hetero\u2010hexamer with four copies of Mis18\u03b1 and two copies of Mis18\u03b2, while the Mis18 holo\u2010complex is a hetero\u2010octamer with two additional copies of Mis18BP1. We identify two conserved consensus Cdk1 phosphorylation sites (T40 and S110) within Mis18BP120\u2013130 and show that phospho\u2010mimicking mutations of these residues disrupt Mis18 complex formation. This explains the molecular basis for Cdk1\u2010mediated timing of Mis18 assembly and CENP\u2010A deposition.Here, we show that Mis18BP1, through its highly conserved N\u2010terminal region comprising amino acids 20\u2013130 integrated in a chromosome arm fl) as well as two different N\u2010terminal fragments: mCherry\u2010Mis18BP120\u2013130 and mCherry\u2010Mis18BP1336\u2013483 (covering the SANTA domain) to the tethering site. Mis18\u03b1 tethered to the ectopic alphoidtetO array recruited Mis18BP1fl and Mis18BP120\u2013130 more robustly compared to the Mis18BP1336\u2013483 and SANT (residues 878\u2013927) domains Figs A. It has\u2013130 Fig B. These 20\u2013130 can directly interact with Mis18\u03b1/\u03b2 in vitro, individually purified recombinant proteins (Mis18\u03b1/\u03b2 and Mis18BP120\u2013130) were analyzed using size\u2010exclusion chromatography (SEC) before and after complex formation. While Mis18\u03b1/\u03b2 and Mis18BP120\u2013130 eluted at 11.0 and 15.7 ml, respectively, the complex containing Mis18\u03b1/\u03b2 and Mis18BP120\u2013130 eluted at 10.9 ml and a C\u2010terminal \u03b1\u2010helical domain and Mis18BP120\u2013130 (12.7 kDa) migrate similarly in the SDS\u2013PAGE . While His\u2010GFP\u2010Mis18\u03b1MeDiY on its own eluted at 13.8 ml, the His\u2010GFP\u2010Mis18\u03b1MeDiY/Mis18BP120\u2013130 complex eluted at 13.3 ml showed that they elute at distinct elution volumes, 13.9 and 12.2 ml, respectively, demonstrating a lack of interaction . Consistent with the SEC analysis shown in Figs MeDiY could bind Mis18BP120\u2013130. His\u2010GFP\u2010Mis18\u03b2MeDiY failed to interact with Mis18BP120\u2013130 even under a low ionic strength condition . The measured MW of the untagged Mis18\u03b1/\u03b2 complex was 151.2 \u00b1 2.9 kDa and correlated well with the calculated MW of a hetero\u2010hexamer , it is almost impossible to accurately determine the subunit stoichiometry within the Mis18 hetero\u2010hexamer. Hence, we introduced a noticeable size variation by purifying His\u2010GFP\u2010Mis18\u03b1 in a complex with His\u2010Mis18\u03b2. Interestingly, the measured MW of His\u2010GFP\u2010Mis18\u03b1/His\u2010Mis18\u03b2 complex was 287.2 \u00b1 5.5 kDa, matching the calculated MW of a hetero\u2010hexamer containing four copies of His\u2010GFP\u2010Mis18\u03b1 and two copies of His\u2010Mis18\u03b2 and His\u2010MBP\u2010Mis18\u03b2C\u2010term (Mis18\u03b2 184\u2010end), we reconstituted the C\u2010terminal helical assembly and analyzed their composition using SEC\u2010MALS by SEC\u2010MALS (175.4 \u00b1 3.3 kDa) is in agreement with the calculated MW of a hetero\u2010octamer containing a Mis18\u03b1/\u03b2 hetero\u2010hexamer plus two copies of Mis18BP120\u2013130 matches the calculated MW at the MeDiY dimeric interface of Mis18\u03b1/\u03b2 and L136A/Y152A/C167A/S169K (Mis18\u03b1PocketM) in the putative substrate\u2010binding pocket using a S. pombe\u2010based homology\u2010modeled structure , His\u2010GFP\u2010Mis18\u03b1DimerM/His\u2010Mis18\u03b2 (Mis18\u03b1DimerM/\u03b2), and His\u2010GFP\u2010Mis18\u03b1PocketM/His\u2010Mis18\u03b2 (Mis18\u03b1PocketM/\u03b2) complexes using a Ni\u2010NTA pull\u2010down assay. While Mis18\u03b1wt/\u03b2 and Mis18\u03b1PocketM/\u03b2 interacted with Mis18BP120\u2013130, Mis18\u03b1DimerM/\u03b2 failed to do so efficiently . This strengthens the notion that the Mis18\u03b1/\u03b2 hetero\u2010trimer is the core oligomeric unit formed through the interactions of the C\u2010terminal helices which then assemble into a hetero\u2010hexamer via the MeDiY\u2010dimerization interface or in combination (Mis18BP120\u2013130 T40E/S110D), on Mis18 complex formation using SEC. Both Mis18BP120\u2013130 T40E and Mis18BP120\u2013130 S110D interacted with Mis18\u03b1/\u03b2. However, Mis18BP120\u2013130 T40E/S110D failed to form a complex with Mis18\u03b1/\u03b2 was recruited by TetR\u2010eYFP\u2010Mis18\u03b1 to the array throughout the cell cycle. Consistent with this, recombinantly purified Mis18BP120\u2013130 T40A/S110A interacted with Mis18\u03b1/\u03b2 as analyzed by SEC . While mutating both these amino acid residues to phospho\u2010mimicking residues completely abolished the ability of Mis18BP1 to bind Mis18\u03b1/\u03b2 in vitro and in vivo, individual phospho\u2010mimic mutants (T40E or S110D) failed do so efficiently. This suggests that Mis18BP1 binds Mis18\u03b1/\u03b2 possibly via a bipartite binding mode. Overall, our findings together with the recent independent study by Pan et alThe temporal regulation of Mis18 complex formation defines the timing of HJURP\u2010mediated CENP\u2010A deposition essential for centromere inheritance and function imer Fig A. HoweveMeDiY, Mis18\u03b1C\u2010term, Mis18\u03b2, Mis18\u03b2MeDiY, and Mis18\u03b2C\u2010term were amplified from codon\u2010optimized sequences (GeneArt) while Mis18BP120\u2013130 was amplified from a human cDNA library . Amplifications were then cloned into pET His6 msfGFP TEV, pET His6 TEV, pET His6 Sumo TEV, pET His6 MBP TEV, pGEX\u20106P\u20101 , pEC\u2010K\u20103C\u2010His\u2010GST, and pEC\u2010K\u20103C\u2010His LIC vectors. pET His6 msfGFP TEV (9GFP Addgene plasmid # 48287), pET His6 TEV (9B Addgene plasmid # 48284), pET His6 Sumo TEV (14S Addgene plasmid # 48291), and pET His6 MBP TEV (9C Addgene Plasmid #48286) were a gift from Scott Gradia.Human Mis18\u03b1, Mis18\u03b1DimerM; V82E/Y176D) and putative substrate\u2010binding pocket mutant and Mis18BP1 mutants, Mis18BP120\u2013130 T40E, Mis18BP120\u2013130 S110D, Mis18BP120\u2013130 T40E/S110D, and Mis18BP120\u2013130 T40A/S110A were generated following the Quikchange site\u2010directed mutagenesis protocol (Stratagene).Mis18\u03b1 MeDiY dimer\u2010disrupting mutant (Mis18\u03b1E. coli BL21 gold grown in LB media. Cultures were induced with 0.35 mM IPTG at 18\u00b0C overnight. Cell lysis was carried out by sonicating cells re\u2010suspended in a lysis buffer containing 20 mM Tris (pH 8.0), 250 mM NaCl (or 500 mM for Mis18BP120\u2013130), 35 mM imidazole, and 2 mM \u03b2ME. Lysis buffer was supplemented with 10 \u03bcg/ml DNase, 1 mM PMSF, and cOmplete . Proteins were purified from the clarified lysates by affinity chromatography using a 5 ml HisTrap HP column . The protein\u2010bound resin was washed with lysis buffer, followed by a buffer containing 20 mM Tris (pH 8.0), 1 M NaCl, 50 mM KCl, 10 mM MgCl2, 2 mM ATP, 35 mM imidazole, 2 mM \u03b2ME, and with a final lysis buffer wash. Proteins were eluted using a lysis buffer supplemented with 500 mM imidazole and dialyzed overnight into 20 mM Tris (pH 8.0), 75\u2013100 mM NaCl, and 2 mM DTT. All proteins were subjected to anion exchange chromatography using the HiTrap Q column . Appropriate fractions were pooled, concentrated, and injected into a Superdex 200 increase 10/300 or Superdex 75 10/300 column equilibrated with 20 mM Tris (pH 8.0), 100\u2013250 mM NaCl, and 2 mM DTT. Fractions were analyzed on SDS\u2013PAGE and stained with Coomassie blue.Proteins were expressed, either individually or together with their binding partners, using 20\u2013130 and made up to 200 \u03bcl with buffer before incubated for 30 min at 4\u00b0C with 60\u2013120 \u03bcl of slurry that had been equilibrated in buffer. Beads were then washed four times with 1 ml of buffer, and bound protein was eluted by boiling in SDS\u2013PAGE loading dye before being analyzed by SDS\u2013PAGE.Ni\u2010NTA pull\u2010down assay was performed in 20 mM Tris (pH 8.0), 75\u2013500 mM NaCl, 10% glycerol, 0.5% NP\u201040, 35 mM imidazole, and 2 mM \u03b2ME. Proteins were mixed with two times molar excess of Mis18BP1280 nm were analyzed by a homo\u2010polymer model using the following parameters for: \u2202A280 nm/\u2202c = 0.97 AU ml mg\u22121 (Mis18\u03b1/\u03b2 complex), 0.86 AU ml mg\u22121 (His\u2010GFP\u2010Mis18\u03b1/His\u2010Mis18\u03b2 and His\u2010Mis18\u03b1/His\u2010GFP\u2010Mis18\u03b2), 0.82 AU ml mg\u22121 (Mis18\u03b1/Mis18\u03b2/Mis18BP120\u2013130), 0.71 AU ml mg\u22121 (Mis18\u03b1/Mis18\u03b2/His\u2010Sumo\u2010Mis18BP120\u2013130), 1.10 AU ml mg\u22121 (His\u2010GFP\u2010Mis18\u03b1C\u2010term/His\u2010MBPMis18\u03b2C\u2010term), 0.86 AU ml mg\u22121 (His\u2010GFP\u2010Mis18\u03b12M/His\u2010Mis18\u03b2), \u2202n/\u2202c = 0.185 ml g\u22121, and buffer RI value of 1.335. The mean standard error in the mass accuracy determined for a range of protein\u2013protein complexes spanning the mass range of 6\u2013600 kDa is \u00b11.9%.Size\u2010exclusion chromatography coupled to UV, static light scattering, and refractive index detection was used to determine the absolute molecular mass of proteins and protein complexes in solution. Injections of 100 \u03bcl of about 1 mg/ml material were run on a Superdex 200 10/300 GL size\u2010exclusion column pre\u2010equilibrated in 50 mM HEPES (pH 8.0), 150 mM NaCl, and 1 mM TCEP at 22\u00b0C with a flow rate of 0.5 ml/min. Light scattering, refractive index (RI), and Afl, Mis18BP120\u2013130, and Mis18BP1336\u2013483 were amplified from a human cDNA library and cloned into the pCDNA3\u2010mCherry or \u2010mCerulean LIC cloning vectors . Mis18\u03b1 was cloned into the TetR\u2010eYFP\u2010IRES\u2010Puro vector. Mutant constructs, TetR\u2010eYFP\u2010Mis18\u03b1DimerM, mCherry\u2010Mis18BP120\u2013130 T40E, mCherry\u2010Mis18BP120\u2013130 S110D, mCherry\u2010Mis18BP120\u2013130 T40E/S110D, and mCherry\u2010Mis18BP120\u2013130 T40A/S110A, were generated using the Quikchange mutagenesis protocol (Stratagene).Human Mis18\u03b2, Mis18BP1tetO array) integrated in a chromosome arm, was maintained in DMEM (Gibco) supplemented with 5% fetal bovine serum (Biowest) and penicillin/streptomycin (Gibco). Cells were grown at 37\u00b0C and 5% CO2. Transfections were performed in parallel with XtremeGene\u20109 (Roche) following manufacturer's instructions. Briefly, 24 h after plating, cells grown in 12\u2010well plates were incubated with transfection complexes containing: 0.25 \u03bcg of each vector, 100 \u03bcl of Opti\u2010MEM (Invitrogen), and 3 \u03bcl of XtremeGene\u20109 reagent for 36 h.The HeLa 3\u20108 overexpressing CENP\u2010A\u2010SNAP integration cell line, containing a synthetic \u03b1\u2010satellite DNA array integration with tetO sites for 5 min at room temperature and blocked with permeabilization buffer containing 3% BSA for 1 h at 37\u00b0C. Mouse anti\u2010CENP\u2010A and Cy\u20105\u2010conjugated donkey anti\u2010mouse secondary antibody (Jackson Immunoresearch) were used for centromere staining.z\u2010stacks were deconvolved using SoftWorx and analyzed using ImageJ software . For CENP\u2010A signal quantification, a custom\u2010made macro in ImageJ modified from Bodor et alTetO array was determined for every z\u2010section within a 7\u2010square pixel box. The mean signal intensity in the array was obtained and the background subtracted using the minimum intensity values within the section. The average intensity of the CENP\u2010A signal from endogenous centromeres was used to normalize. At least three biological independent experiments were performed for each assay.Micrographs were acquired in the Centre Optical Instrumentation Laboratory on a DeltaVision Elite system (Applied Precision) using an inverted Olympus IX\u201071 stand, with an Olympus UPlanSApo 100\u00d7 oil immersion objective 1.4) and a Lumencor light source. Camera (Photometrics Cool Snap HQ), shutter and stage were controlled through SoftWorx (Applied Precision). The 0.2\u2010\u03bcm\u2010spaced AAJ conceived the project. FS, BM\u2010P, MAA, OM, AAJ, and WCE designed experiments. FS, BM\u2010P, MAA, MAW, and OM performed experiments and analyzed data. FS, BM\u2010P, MAA, OM, WCE, and AAJ wrote the manuscript.The authors declare that they have no conflict of interest.Expanded View Figures PDFClick here for additional data file.Review Process FileClick here for additional data file."} +{"text": "The crystal structure of 2-(5-bromo\u00adthio\u00adphen-2-yl)aceto\u00adnitrile, previously reported as a liquid, has short centrosymmetric Type I Br\u22efBr halogen inter\u00adactions. 6H4BrNS, crystallizes in the space group P21/n with one complete mol\u00adecule in the asymmetric unit. The non-H atoms are nearly planar (r.m.s for non-H atoms = 0.071\u2005\u00c5), with the nitrile group oriented anti\u00adperiplanar with respect to the thio\u00adphene S atom. Inter\u00admolecular Type I centrosymmetric Br\u22efBr halogen inter\u00adactions are present at a distance of 3.582\u2005(1)\u2005\u00c5 and with a C\u2014Br\u22efBr angle of 140.7\u2005(1)\u00b0. Additional weaker C\u2014H\u22efN, C\u2014H\u22efS, and S\u22ef\u03c0 inter\u00adactions are also present. A Hirshfeld analysis indicates Br\u22efBr inter\u00adactions comprise only 1.9% of all the inter\u00adatomic contacts.The title compound, C The large number of reflections in the data sets (and the Fourier-transform relationship of intensities to atoms) ensures that no particular bias was thereby introduced.10.1107/S2056989018000968/dx2004sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018000968/dx2004Isup2.hklStructure factors: contains datablock(s) I. DOI: 1817195CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the two structures, short cationic tris\u00adtannatoxanes (C 3(CH3)9(OH)2(H2O)2][Sn(CH3)3(CHO2)2] (1) and [Sn3(CH3)9(OH)2]Cl\u00b7H2O (2), are partially condensed products of hydrolysed tri\u00admethyl\u00adtin chloride. In the structures of 1 and 2, short cationic tris\u00adtannatoxanes (C9H29O2Sn3) are bridged by a diformatotri\u00admethyl\u00adtin anion or a chloride anion, respectively. Hydrogen bridges are present and supposedly stabilize these structures against further polymerization to the known polymeric tri\u00admethyl\u00adtin hydroxide. Especially noteworthy is that the formate present in this structure was formed from atmospheric CO2.The title compounds, [Sn Therefore, the activation of CO2 plays an important role in today\u2019s research. It is already known that CO2 is activated by electroreduction of different metals \u00b0 while in 2 it is 135.30\u2005(17)\u00b0. In the chloride structure 2, a change in two further angles is noticed. The O1\u2014Sn1\u2014Cl1 angle [177.58\u2005(10)\u00b0] and the O2\u2014Sn3\u2014Cl1\u2032 angle [175.5\u2005(12)\u00b0] decreases and the chloride anion.In the crystal structures, no polymeric Sn\u2013O structures were formed, as found in the tri\u00admethyl\u00adtin hydroxide. The short tri\u00admethyl\u00adtin hydroxide chain has a positive and the chloride or bisformatostannate a negative charge. In the structure of via hydrogen bonds. In structure 1 , thus forming a two-dimensional network. Additionally, hydrogen bonds between these sheets form a two-dimensional network along the bc plane (O4\u22efH1\u2014O1).As described, both structures are inter\u00admolecularly linked 1 Fig.\u00a03, the for2 \u2005\u00c5], one by a Cl1\u22efH1i\u2014O1i hydrogen bond [3.251\u2005(4)\u2005\u00c5]. A fourth hydrogen bond, Cl1\u22efH3ii\u2014O3ii [3.068\u2005(5)\u2005\u00c5], results in a distorted tetra\u00adhedral environment. Thus, a three-dimensional network of hydrogen bridges is formed. The inter\u00adactions between Sn\u2013Cl differ due to steric repulsion of the C2 and C7iii methyl groups. The van der Waals radius of a methyl group is 2\u2005\u00c5 \u00b0 for Sn1\u2014Cl1\u2014Sn2 and 179.2\u2005(2)\u00b0 for O1\u2014Sn1\u2014Cl1 were observed = 1.5Ueq(Cmeth\u00adyl). The CH3 hydrogen atoms were allowed to rotate but not to tip. Due to point group symmetry 2 of both the cation and anion in 1, with the twofold rotation axis running through the respective central Sn atom and one of the methyl groups, the latter is equally disordered over two positions.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016014912/su5326sup1.cifCrystal structure: contains datablock(s) Global, 1, 2. DOI: 10.1107/S2056989016014912/su53261sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989016014912/su53262sup3.hklStructure factors: contains datablock(s) 2. DOI: 1505529, 1505528CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Two new crystal structures of eight- and ten-membered cyclic bis\u00adanisyl\u00adphosphono\u00adthioyl disulfanes have been determined and these are compared to the structures of their ferrocenyl analogues. 5,5\u03bb5-dioxadi\u00adthiadi\u00adphospho\u00adcane-2,5-di\u00adthione, C16H18O4P2S4, and 2,5-bis\u00ad(4-meth\u00adoxy\u00adphen\u00adyl)-1,6,3,4,2\u03bb5,5\u03bb5-dioxadi\u00adthia\u00addiphosphecane-2,5-di\u00adthione, C18H22O4P2S4, have been determined and compared to structures of the ferrocenyl analogues. The eight-membered rings have similar conformations (TBC) but the ten-membered macrocycles are differently puckered. Structural parameters of the relevant SPSSPS motif have been analysed and are discussed in detail. Compound 1 was refined as an inversion twin and 2 was refined as a two-component rotational twin.Two new crystal structures of eight- and ten-membered cyclic bis\u00adanisyl\u00adphosphono\u00adthioyl disulfanes, namely 2,5-bis\u00ad(4-meth\u00adoxy\u00adphen\u00adyl)-1,6,3,4,2\u03bb Lawesson reagent LR P(S)SH and Fc(RO)P(S)SH, obtained in a simple reaction between LR or fLR and alcohols, are of considerable inter\u00adest because they form a plethora of structurally inter\u00adesting chelate complexes with metal ions S]e.g. eight-membered 1a acid derivatives and among them the unique eight-, nine- and ten-membered cyclic bis\u00adanisyl\u00adphosphono\u00adthioyl disulfanes . The related ferrocenyl compound 1a crystallizes in space group C2/c with non-typical three and half independent mol\u00adecules in the asymmetric unit (Z = 28), which complicates comparisons.Views of mol\u00adecular structures and atom-labeling scheme for 2 forms a monoclinic crystalline phase obeying P21/c space-group symmetry with one mol\u00adecule in the asymmetric unit and Z = 4. The related ferrocenyl structure 2a crystallizes in space group PCompound trans arrangement, i.e. above and below the macrocycle ring plane for all compounds 1\u20132a, which is also typical for all open-chain bis\u00adphospho\u00adrothioyl disulfanes studied previously , 2.0704\u2005(10), 2.0685\u2005(10), 2.0711\u2005(15)\u2005\u00c5 for 1a; 2.074\u2005(3)\u2005\u00c5 for 2 and 2.0788\u2005(9)\u2005\u00c5 for 2a. They are longer than the typical S\u2014S bond lengths for known diorganyl disulfanes RSSR [2.05\u2005(3)\u2005\u00c5]. The observed S\u2014S bond elongation in 1\u20132a may be correlated with the PSSP torsion angles are shorter than the endocyclic P\u2014S bonds (ca 2.10\u2005\u00c5).The S\u2014S bond lengths have values of 2.068\u2005(2)\u2005\u00c5 for 1\u20132a adopt a distorted tetra\u00adhedral geometry, where the C\u2014P=S angles deviated the most (116.1\u2013118.5\u00b0) from the ideal tetra\u00adhedral angle. This is obviously due to the steric effects of the anisyl and ferrocenyl substituents. On the other hand, it is worthy to note that the O\u2014P\u2013S bond angles in 1\u20132a (107\u2013108\u00b0) are not distorted, probably due to minimal conformational strain present in those medium-sized heterocycles. Moreover, both the P=S and aromatic anisyl groups in 1 are almost perfectly coplanar (unlike P=S and the cyclopentadienyl ring in 1a), which provides energetically favorable conjugation [torsion angle S2\u2014P1\u2014C10\u2014C15 = \u22123.8\u2005(4)\u00b0 in 1vs 35.75\u2005(3)\u00b0 for the equivalent angle in a selected representative mol\u00adecule with Fe7 in 1a]. The other related independent torsion angles in 1a are \u221231.\u2005(3), \u221233.9\u2005(3), \u221227.0\u2005(3), \u221228.7\u2005(3), 34.8\u2005(3), 35.7\u2005(3)\u00b0, for Fe1\u2013Fe6, respectively.All phospho\u00adrus atoms in 1 is the most symmetric with the lowest PSSP torsion [\u221293.68\u2005(8)\u00b0] and shows only a moderate deviation from a right angle. The PSSP torsion angles in 1a are 6\u20138\u00b0 wider than in 1. Notably, ten-membered disulfanes have even wider PSSP torsion angles and the difference between them is smaller, \u2212112.89\u2005(11) and 114.9\u2005(4)\u00b0, for 2 and 2a, respectively.It is well recognised that PSSP torsion is a characteristic feature of all disulfanes as a class of organic compounds. The structure of X (X = O or S) can be found in the structures of 1 and 2 , 4.2670\u2005(9), 4.2652\u2005(9) or 4.261\u2005(1)\u2005\u00c5 (for different independent mol\u00adecules in 1a) for eight-membered rings, to 4.614\u2005(2) in 2 and 4.604\u2005(1)\u2005\u00c5 in 2a for the ten-membered rings.The transannular P\u22efP distances are very similar within the same ring size and increase, from 4.3331\u2005(17)\u2005\u00c5 in 1 and 1a was recognised by PLATON \u2005\u00c5. The anisyl substituents may have inhibited this kind of inter\u00adaction.The strongest inter\u00admolecular hydrogen-bonding inter\u00adaction in 2 are mainly based on the anisyl methoxyl CH3O oxygen atoms O3 and O4 and the P=S sulfur atom S3 as acceptors. Hydrogen-bond donors are the anisyl ortho-hydrogen atoms or methyl\u00adene hydrogen atoms. Moreover, some C\u2014H..\u03c0. inter\u00adactions may play some role in the system, e.g. C16\u2014H16A\u22efring(C20\u2013C25), see Fig.\u00a05Inter\u00admolecular inter\u00adactions in et al., 2016et al., 2015et al., 2017et al., 2004et al., 1991et al., 1993Bisphosphono\u00adthioyl disulfanes represent a rather rare class of compounds solvent system.Eight- and ten-membered cyclic bis\u00adanisyl\u00adphosphono\u00adthioyl disulfanes 2,5-Bis(4-meth\u00adoxy\u00adphen\u00adyl)-1,6,3,4,2,5-dioxadi\u00adthiadi\u00adphos\u00adpho\u00adcane 2,5-di\u00adthione, 1M.p. 441-443\u2005K.2,5-Bis(4-meth\u00adoxy\u00adphen\u00adyl)-1,6,3,4,2,5-dioxadi\u00adthia\u00addiphos\u00adphecane 2,5-di\u00adthione, 2Yield: 65%, m.p. 415\u2013417\u2005K.1H NMR (CDCl3): 2.20 , 2.25 , 3.89 , 4.37 , 4.89 , 7.01 , 7.87 .13C NMR: 27.21 , 55.46 (s), 67.08 , 114.03 , 125.41 , 132.89 , 163.09 (s).31P{1H} NMR (CDCl3): 89.19 (3JPP = 4\u2005Hz)18H22O4P2S4: 492.0. Found: 492.9 [M+H]+.MS calculated for C1 was refined as an inversion twin with contribution of the second domain equal to 0.45\u2005(17). This explains the ambiguous Flack parameter and is not surprising since we started from achiral substrates. Structure 2 was refined as a two-component rotational twin with twin law: {2 (Q1\u2013Q3 ca 2e \u00c53), which are close to sulfur atoms , may stem from conformational flexibility of the ring. Note: the structure of 1 was determined at room temperature (due to a failure of our CryoStream unit) not at 120\u2005K as for 2 but we believe it did not influence the qualitative conclusions drawn from the results.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018001068/zp2026sup1.cifCrystal structure: contains datablock(s) 2, global, 1. DOI: 10.1107/S2056989018001068/zp20261sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989018001068/zp20262sup3.hklStructure factors: contains datablock(s) 2. DOI: Click here for additional data file.10.1107/S2056989018001068/zp20261sup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018001068/zp20262sup5.cmlSupporting information file. DOI: 1558043, 719124CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, a pheno\u00adthia\u00adzine moiety is linked to a planar carbazole unit (r.m.s. deviation = 0.029\u2005\u00c5) by a C\u2014C single bond. Their mean planes are inclined to one another by 27.28\u2005(5)\u00b0, and the pheno\u00adthia\u00adzine moiety possesses a typical butterfly structure with a fold angle of 27.36\u2005(9)\u00b0 between the two benzene rings. 29H24N2OS, contains a pheno\u00adthia\u00adzine moiety linked to a planar carbazole unit (r.m.s. deviation = 0.029\u2005\u00c5) by a C\u2014C single bond. The pheno\u00adthia\u00adzine moiety possesses a typical non-planar butterfly structure with a fold angle of 27.36\u2005(9)\u00b0 between the two benzene rings. The dihedral angle between the mean planes of the carbazole and pheno\u00adthia\u00adzine units is 27.28\u2005(5)\u00b0. In the crystal, mol\u00adecules stack in pairs along the c-axis direction, linked by offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.797\u2005(1)\u2005\u00c5]. There are C\u2014H\u22ef\u03c0 inter\u00adactions present linking these dimers to form a three-dimensional structure.The title compound, C The fold angle of 27.36\u2005(9)\u00b0 between the two benzene rings of this moiety compares well with the values reported for similar compounds \u2005\u00c5, inter\u00adplanar distance = 3.5133\u2005(8)\u2005\u00c5, slippage = 1.439\u2005\u00c5, Cg5 is the centroid of the C15\u2013C20 ring; symmetry code: (i) \u2212x\u00a0+\u00a01, \u2212y, \u2212z\u00a0+\u00a02]. There are also C\u2013H\u22ef\u03c0 inter\u00adactions present linking these dimers to form a three-dimensional structure -10H-pheno\u00adthia\u00adzine-3-carbaldehyde , 9-ethyl-9H-carbazole-3-boronic acid pinnacol ester , Pd(PPh3)4 and K2CO3 under high vacuum was added a mixture of toluene:water (2:1). The resulting mixture was heated to reflux under an N2 atmos\u00adphere for ca 24\u2005h. On completion of the reaction (monitored by TLC), it was quenched by addition of saturated double-distilled H2O and extracted with di\u00adchloro\u00admethane. The organic phases were collected and washed with brine and dried over anhydrous Na2SO4 and then concentrated. The product was purified by column chromatography on silica gel using ethyl acetate:n-hexane as eluent, to give the title compound as a pale-yellow crystalline solid (yield 80%). It was characterized by 1H NMR, 13C NMR, IR and ESI\u2013MASS. Brown block-like crystals of the title compound were obtained by slow evaporation at room temperature of a solution in di\u00adchloro\u00admethane and aceto\u00adnitrile (1:1 v/v).To a mixture of 7-bromo-10-ethyl-10Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017005540/su5357sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017005540/su5357Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017005540/su5357Isup3.cmlSupporting information file. DOI: 1543611CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming chains propagating along the 14H15NO4S, the di\u00adhydro\u00adthia\u00adzine ring adopts a distorted sofa conformation with the S atom displaced from the mean plane through the N and C ring atoms by 0.767\u2005(1)\u2005\u00c5. The allyl substituent (C\u2014C=C) is inclined to this mean plane by 78.5\u2005(7)\u00b0 and the acetate group [C(=O)\u2014O\u2014C] by 66.5\u2005(3)\u00b0. In the crystal, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions forming chains propagating along the a-axis direction.In the title compound, C The allyl substituent (C\u2014C=C) is inclined to this mean plane by 78.5\u2005(7)\u00b0 and the acetate group (O=C\u2014O\u2014C) by 66.5\u2005(3)\u00b0. Atom N1 has a planar configuration, the sum of the bond angles being 359.1\u00b0.The mol\u00adecular structure of the title compound, A\u22efC5 = 2.47\u2005\u00c5 in this fragment [the sum of the van der Waals radii \u00b0]. The allyl group is orthogonal to the benzo\u00adthia\u00adzine fragment plane while the terminal double bond is synperiplanar to the N1\u2014C12 bond . The steric repulsion between the allyl substituent and the aromatic cycle (short intra\u00admolecular contacts H2\u22efC12 = 2.77\u2005\u00c5 and H12A\u22efC2 = 2.83\u2005\u00c5) results in the elongation of the C1\u2014N1 bond [1.411\u2005(5)\u2005\u00c5], compared with the mean value of 1.371\u2005\u00c5 of K2CO3 and the mixture was stirred for 30\u2005min. Allyl bromide was then added and the mixture was stirred for a further 30\u2005min at 298\u2005K. It was then diluted with cold water and acidified with dilute HCl to pH 4. It was extracted with CH2Cl2 (3 \u00d7 10\u2005ml). The organic extracts were combined and the solvent removed by distillation (at reduced pressure at the end). The residue was dissolved in 20\u2005ml of hot methanol and filtered over charcoal. The resulting solution was then placed in a freezer (253\u2005K) for 24\u2005h, after which crystals of the title compound were harvested .The synthesis of the title compound, Uiso = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016015978/su5329sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989016015978/su5329Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016015978/su5329Isup3.cmlSupporting information file. DOI: 1508990CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atom in the title complex, [Cd(C10H14N2O)2(C8H4NO2)2(H2O)2], is located on an inversion centre and is coordinated by an N2O4 donor set from pairs of water, 4-cyano\u00adbenzoate and N,N-di\u00adethyl\u00adnicotinamide ligands.The Cd 10H14N2O)2(C8H4NO2)2(H2O)2], is centrosymmetric and contains two water mol\u00adecules, two 4-cyanobenzoate (CB) ligands and two di\u00adethyl\u00adnicotinamide (DENA) ligands. All the ligands are coordinated to the CdII atom in a monodentate mode. The four nearest O atoms around the CdII atom form a slightly distorted square-planar arrangement, with the distorted octa\u00adhedral coordination sphere being completed by the two pyridine N atoms of the DENA ligands at distances of 2.3336\u2005(13)\u2005\u00c5. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 8.75\u2005(16)\u00b0, while the benzene and pyridine rings are oriented at a dihedral angle of 57.83\u2005(5)\u00b0. The water mol\u00adecules exhibit both intra\u00admolecular and inter\u00admolecular O\u2014H\u22efO hydrogen bonds. The latter lead to the formation of supra\u00admolecular chains propagating along [110].The mononuclear title cadmium complex, [Cd(C N,N\u2032-di\u00adethyl\u00adnicotinamide (DENA) is an important respiratory stimulant 2(C7H4ClO2)2(H2O)2] 2(H2O)2] is one form of niacin. A deficiency of this vitamin leads to loss of copper from the body, known as pellagra disease. Pellagra patients show unusually high serum and urinary copper levels and DENA ligands, namely trans-di\u00adaqua\u00adbis\u00ad(4-cyano\u00adbenzoato-\u03baO)bis\u00adcadmium, [Cd(CB)2(DENA)2(H2O)2], and report herein its crystal structure.The structure\u2013function\u2013coordination relationships of the aryl\u00adcarboxyl\u00adate ion in CdII atom located on an inversion centre, one CB ligand, one DENA ligand as well as one water mol\u00adecule, all ligands coordinating to the CdII atom in a monodentate mode of the two symmetry-related monodentate CB anions and water O atoms (O4 and O4i) form a slightly distorted square-planar arrangement around the Cd1 atom, while the slightly distorted octa\u00adhedral coordination sphere is completed by the two pyridine N atoms (N1 and N1i) of two DENA ligands \u2005\u00c5]; the Cd\u2014N bond length is the longest with 2.3336\u2005(13)\u2005\u00c5 in the CdO4N2 octa\u00adhedron. The Cd1 atom lies 0.7558\u2005(1)\u2005\u00c5 below the planar (O1/O2/C1) carboxyl\u00adate group. The O\u2014Cd\u2014O and O\u2014Cd\u2014N bond angles range from 87.54\u2005(5) to 92.46\u2005(5)\u00b0. In the carboxyl\u00adate groups, the C\u2014O bonds of the coordinating O atoms [C1\u2014O1 = 1.244\u2005(2)\u2005\u00c5 and C1\u2014O2 = 1.259\u2005(2)\u2005\u00c5] are 0.015\u2005(2)\u2005\u00c5 longer than those of the non-coordinating ones, indicating delocalized bonding arrangements rather than localized single and double bonds. The dihedral angle between the carboxyl\u00adate group (O1/O2/C1) and the adjacent benzene (C2\u2013C7) ring is 8.75\u2005(16)\u00b0, while the benzene and pyridine (N1/C9\u2013C13) rings are oriented at a dihedral angle of 57.83\u2005(5)\u00b0.The two carboxyl\u00adate O atoms hydrogen bonds (Table\u00a01S(6) hydrogen-bonding motifs hydrogen bonds in H2O (50\u2005ml) and di\u00adethyl\u00adnicotinamide in H2O (10\u2005ml) with sodium 4-cyano\u00adbenzoate in H2O (100\u2005ml). The mixture was filtered and set aside to crystallize at ambient temperature for several days, giving colourless single crystals.The title compound was prepared by the reaction of CdSO2O) were located in a difference Fourier map and were refined freely. The C-bound H atoms were positioned geometrically with C\u2014H = 0.93, 0.97 and 0.96\u2005\u00c5, for aromatic, methyl\u00adene and methyl H atoms, respectively, and constrained to ride on their parent atoms, with Uiso(H) = k \u00d7 Ueq(C), where k = 1.5 for methyl H atoms and k = 1.2 for aromatic and methyl\u00adene H-atoms.Experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0210.1107/S2056989016018247/wm5339sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016018247/wm5339Isup2.hklStructure factors: contains datablock(s) I. DOI: 1517222CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the title compound comprises one cafenstrole ligand mol\u00adecule and one silver nitrate ion. The coordination bonds between silver and oxygen atoms allow a continuous one-dimensional coordination polymer structure along [001]. The three-dimensional architecture is stabilized by C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions N,N-diethyl-3-mesitylsulfonyl-1H-1,2,4-triazole-1-carboxamide), a triazole herbicide, leads to the title coordination polymer, [Ag(NO3)(C16H22N4O3S)]n, whose asymmetric unit comprises one cafenstrole ligand mol\u00adecule, one AgI atom and one nitrate ion. The AgI atom, with a distorted trigonal\u2013pyramidal environment, is coordinated by one nitro\u00adgen atom of a triazole ring, two oxygen atoms of a nitrate ion and one oxygen atom of a neighboring nitrate ion. The coordination bonds between silver and oxygen atoms give rise to a one-dimensional (1D) coordination polymer structure along [001]. The dihedral angle between the planes of the triazole and benzene rings is 87.13\u2005(11)\u00b0. In the crystal, the coordination polymer is stabilized by C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, resulting in a three-dimensional architecture.The reaction of silver nitrate and cafenstrole ( L; Kang et al., 2015et al., 2001et al., 2013et al., 2014Recently, we have reported the crystal structure of the ligand cafenstrole [Ag1\u2014N1 = 2.250\u2005(3)\u2005\u00c5] and three oxygen atoms . The Ag1, O5, N1, and O5i atoms form a slightly distorted triangular basal plane with bond angles O5\u2014Ag1\u2014O5i = 106.52\u2005(5), O5\u2014Ag1\u2014N1 = 118.75\u2005(11) and O5i\u2014Ag1\u2014N1 = 134.63\u2005(11)\u00b0. The apex atom, O4, deviates considerably from the normal to the basal plane, as indicated by the O4\u2014Ag1\u2014N1 bond angle of 149.66\u2005(10)\u00b0. Other bond angles are 48.93\u2005(10) and 67.18\u2005(10)\u00b0 for O4\u2014Ag1\u2014O5 and O4\u2014Ag1\u2014O5i, respectively. One oxygen atom of the nitrate ion (O6) is not bound to the AgI ion, whereas the other two oxygen atoms of the nitrate ion (O4 and O5) are bound to the AgI ion. One of the bound O atoms (O5) links neighbouring AgI ion ions, thus forming a 1D polymer along [001]. The triazole plane is rotated about the S1\u2013C10 axis in the opposite direction in comparison with free cafenstrol \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, z\u00a0+\u00a01 symmetry, resulting in a 1D chain along [001] inter\u00adactions , was determined by Min et al. 2(NO3)(C12H8N2)]\u00b72C12H8N2, has a polymeric chain structure, where nitrate ions show similar coordination bonds compared to those in the title compound, but with AgI ions replaced by with PrIII atoms in methanol (5\u2005mL). The ligand was purchased from the Dr Ehrenstorfer GmbH Company. Single crystals suitable for X-ray crystallography were obtained by slow evaporation of the solvent at room temperature after one week.The title compound was prepared from a mixed solution of the cafenstrole ligand in acetone (5\u2005mL) and Ag(NOd(C\u2014H) = 0.98\u2005\u00c5, Uiso(H) = 1.5Ueq(C) for methyl group, d(C\u2014H) = 0.99\u2005\u00c5, Uiso(H) = 1.2Ueq(C) for Csp3\u2014H and d(C\u2014H) = 0.95\u2005\u00c5, Uiso(H) = 1.2Ueq(C) for aromatic C\u2014H.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016016662/vn2117sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989016016662/vn2117Isup2.hklStructure factors: contains datablock(s) I. DOI: 1510357CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports6: Article number: 28177; 10.1038/srep28177 published online: 07052016; updated: 10102016.In this Article the legend for Figure 5 is incomplete.2+ in cytosol was measured using Fura-2 and Ca2+ in ER was measured using FRET-based probe D1ER cameleon in the basal state and after thapsigargin stimulation (100\u2009nM for 1000\u2009sec). Representative Ca2+ measurements in cytosol and ER (left panel). Quantitative ER Ca2+ levels of the average value prior to thapsigargin stimulation and the minimum value after addition of thapsigargin (Tg(+)) (right panel). Data are means\u2009\u00b1\u2009SEM, n\u2009=\u200943 for control, n\u2009=\u200950 for IRS1KO, and n\u2009=\u200940 for IRS2KO \u03b2-cells. *P\u2009<\u20090.05, **P\u2009<\u20090.01\u201d.\u201cCaShould read:2+ in cytosol was measured using Fura-2 and Ca2+ in ER was measured using FRET-based probe D1ER cameleon in the basal state and after thapsigargin stimulation (100\u2009nM for 1000\u2009sec). Representative Ca2+ measurements in cytosol and ER (left panel). Quantitative ER Ca2+ levels of the average value prior to thapsigargin stimulation and the minimum value after addition of thapsigargin (Tg(+)) (right panel). Data are means\u2009\u00b1\u2009SEM, n\u2009=\u200943 for control, n\u2009=\u200950 for IRS1KO, and n\u2009=\u200940 for IRS2KO \u03b2-cells. *P\u2009<\u20090.05, **P\u2009<\u20090.01. (b) Immunoblot of SERCA2 and \u03b2-actin in control \u03b2-cells, IRS1KO \u03b2-cells, and IRS2KO \u03b2-cells incubated with vehicle or thapsigargin (100\u2009nM) for 8\u2009h. Data are means\u2009\u00b1\u2009SEM, n\u2009=\u20094. *P\u2009<\u20090.05\u201d.\u201c(a) Ca"} +{"text": "The structures of three bicyclic carbohydrates derivatives containing cyclo\u00adbutanone or cyclo\u00adlactame beside the pyran\u00adose ring are reported and the conformation and configuration established. R,3R,4R,5R,6S)-4,5-bis\u00ad(acet\u00adyloxy)-7-oxo-2-oxabi\u00adcyclo[4.2.0]octan-3-yl]methyl acetate, C14H18O8, (I), [-5-acet\u00adyloxy-7-hy\u00addroxy\u00adimino-2-oxobi\u00adcyclo\u00ad[4.2.0]octan-4-yl acetate, C11H15NO6, (II), and pyrrol-5-yl]methyl acetate, C14H19NO8, (III), are stable bicyclic carbohydrate derivatives. They can easily be synthesized in a few steps from commercially available glycals. As a result of the ring strain from the four-membered rings in (I) and (II), the conformations of the carbohydrates deviate strongly from the ideal chair form. Compound (II) occurs in the boat form. In the five-membered lactam (III), on the other hand, the carbohydrate adopts an almost ideal chair conformation. As a result of the distortion of the sugar rings, the configurations of the three bicyclic carbohydrate derivatives could not be determined from their NMR coupling constants. From our three crystal structure determinations, we were able to establish for the first time the absolute configurations of all new stereocenters of the carbohydrate rings.The title compounds, [(1 The conformation of the pyran\u00adose rings deviates from the ideal chair. The puckering amplitudes and smallest displacement parameters for mol\u00adecules A and B are q = 0.467\u2005(2)/0.473\u2005(3)\u2005\u00c5, \u03b8 = 151.9\u2005(4)/150.7\u2005(4)\u00b0 and \u03c6 = 114.1\u2005(6)/114.3\u2005(6)\u00b0. The main feature is the absolute configuration of the new stereocenters being 1R and 6S. Surprisingly, the acet\u00adyloxy substituents are positioned axially, in contrast to the usual d-gluco arrangement. Obviously the cyclo\u00adbutanone ring, with its bis\u00adectional positioned (C1\u2014C8) and axial bonds (C6\u2014C7) \u2013 in relation to the pyran\u00adose ring \u2013 enforces a flipping of the chair from 4C1 into 1C4. The cyclo\u00adbutane ring is almost planar [maximum deviation from the best plane of C7 = 0.0762\u2005(15)\u2005\u00c5 in A and 0.0815\u2005(15)\u2005\u00c5 in B] and can be described by the dihedral angles, forming by folding along the C6\u22efC8 and C1\u22efC7 line, between the planes C1\u2013C6\u2013C8/C6\u2013C7\u2013C8 and C1\u2013C6\u2013C7/C1\u2013C7\u2013C8 . The deviation of the carbonyl O atoms (O8A/O8B) from the mean plane of the pyran ring are 0.253\u2005(5) and 0.303\u2005(6)\u2005\u00c5 in mol\u00adecules A and B, respectively. The dihedral angles between the pyran\u00adose rings and the cyclo\u00adbutane rings are 61.3\u2005(1) and 62.1\u2005(1)\u00b0 for mol\u00adecules A and B, respectively. Four non-classical intra\u00admolecular hydrogen bonds for each of the both mol\u00adecules can be observed A is disordered . Mol\u00adecules A, B and C mainly differ in the torsion angles C10\u2014O4\u2014C5\u2014C6 and O4\u2014C5\u2014C6\u2014C1 of the acet\u00adyloxy substituents in the 5-position /0.763\u2005(3)/0.75\u2005(3)\u2005\u00c5, \u03b8 = 90.9\u2005(6)/91.0\u2005(2)/91\u2005(2)\u00b0 and \u03c6 = 12.6\u2005(6)/12.7\u2005(2)/28\u2005(3)\u00b0 for mol\u00adecules A, B and C, and not the usual chair conformation. This arrangement is caused by the cyclo\u00adbutane ring with the C1\u2014C8 and C6\u2014C7 bonds, which are bis\u00adectional related to the arabinose ring. The absolute configuration on the stereocenters of the shared ring atoms is C1S and C6R. The cyclo\u00adbutane rings are almost planar with maximum deviations from the best plane of 0.045\u2005(3)\u2005\u00c5 (C7A), 0.039\u2005(1)\u2005\u00c5 (C7B) and 0.072\u2005(12)\u2005\u00c5 (C7C). The nitro\u00adgen atoms deviate marginally from these planes [N1A \u22120.224\u2005(9)\u2005\u00c5, N1B 0.199\u2005(4)\u2005\u00c5, N1C 0.30\u2005(4)\u2005\u00c5. The dihedral angles within the four-membered rings between C1/C6/C8 and C6/C7/C8 are 9.4\u2005(5)\u00b0 (A), 8.2\u2005(2)\u00b0, (B) and 15\u2005(2)\u00b0 (C), and between C1/C6/C7 and C1/C7/C8 they are 9.0\u2005(5)\u00b0 (A), 7.9\u2005(3)\u00b0 (B) and 14\u2005(2)\u00b0 (C). The hydroxyl group of the oxime substit\u00aduent can adopt two different configurations. Mol\u00adecule B exhibits an E configuration. For disordered mol\u00adecules A and C, the E/Z ratio of the isomers is 0.802\u2005(7):0.198\u2005(7). Thus, the major component (A) is E configured, with the hydroxyl group pointing away from the six-membered ring. In the minor Z isomer (C), the hydroxyl group exhibits a sterically unfavourable inter\u00adaction with the carbohydrate ring. An intra\u00admolecular hydrogen bond between C5A/C5C and O5A is observed on Fig.\u00a02. The pyred Fig.\u00a02.S configured. The six-membered and the five-membered rings are fused in the cis configuration. The C3a\u2014C3 bond is axial and the C7a\u2014N1 bond is bis\u00adectionally positioned with respect to the pyran\u00adose ring. The pyran\u00adose ring exhibits a strongly distorted chair conformation, with puckering parameters q = 0.555\u2005(3)\u2005\u00c5, \u03b8 = 20.4\u2005(3)\u00b0 and \u03c6 = 267.9\u2005(9)\u00b0. The usual d-gluco configuration in the chair form 4C1 is found, in contrast to (I)Compound (III)A mol\u00adecules are hydrogen-bonded via C4A\u2014H4A\u22efO3Ai inter\u00adactions screwing around the b-axis direction . The A and B mol\u00adecules are linked by two further hydrogen bonds (C10B\u2014H104\u22efO8Aii and C10B\u2014H106\u22efO7Aii).The crystal packing of (I)on Fig.\u00a04. BetweenA mol\u00adecules are in an alternating arrangement with those consisting only of B mol\u00adecules, both screwing along the b-axis direction on Fig.\u00a05. In contvia weak C\u2014H\u22efO hydrogen bonds running along the a-axis direction. The chains formed this way are connected in a pairwise fashion by strong N1\u2014H1A\u22efO8 bonds along c et al. was synthesized from tri-O-acetyl-d-glucal, commercially available or obtained by the procedure of Ferrier in dry diethyl ether (30\u2005mL) at room temperature over 30\u2005min under an N2 atmosphere. After completion of the reaction (TLC control), zinc dust was added at 273\u2005K. Acetic acid (13\u2005mL) was added within 10\u2005min and the reaction mixture was stirred for 60\u2005min. The reaction mixture was diluted with diethyl ether (60\u2005mL) and the insoluble materials were filtered off through Celite, which was washed with diethyl ether (5 \u00d7 50\u2005mL) and methanol (50\u2005mL). The filtrate was extracted with (3 \u00d7 100\u2005mL) water. The organic layer was dried over MgSO4 and concentrated in vacuo. The resulting residue was purified by column chromatography (hexa\u00adne/ethyl acetate 5:1) to afford pure cyclo\u00adbutanone (I)rier 1965. Tri\u00adchlOxime (II) was synthesized from di-O-acetyl-d-arabinal, obtained by the procedure of Ferrier the corresponding cyclo\u00adbutanone was synthesized as described above and isolated by column chromatography (hexa\u00adne/ethyl acetate 5:1) in 83% yield. 242\u2005mg (1.0\u2005mmol) of this cyclo\u00adbutanone was dissolved in ethanol (2\u2005mL) and then added to a solution of sodium acetate and hydroxyl\u00adamine hydro\u00adchloride in water (2\u2005mL). The reaction mixture was stirred at 327\u2005K for 2\u2005h and then for 1\u2005h at room temperature. The reaction mixture was washed with water (30\u2005mL) and extracted with CH2Cl2 (3 \u00d7 50\u2005mL). The organic layers were combined, dried over MgSO4, filtered and concentrated in vacuo. The oxime (II)rier 1965. StartinLactam (III) was synthesized from cyclo\u00adbutanone (I)2Cl2 (3 \u00d7 50\u2005mL). The organic layers were combined, dried over MgSO4, filtered and concentrated in vacuo. Thionyl chloride was added to a solution of the crude oxime in 1,4-dioxane (4\u2005mL), and stirred for 10\u2005min at room temperature. The reaction was quenched with saturated aqueous NaHCO3 (50\u2005mL), and extracted with EtOAc (3 \u00d7 100\u2005mL). The organic extracts were washed with brine, dried over MgSO4, and concentrated in vacuo. The residue was purified by column chromatography (hexa\u00adne/ethyl acetate 1:4) to afford the lactam in analytically pure form . Single crystals suitable for X-ray diffraction were prepared by slow evaporation of a solution of (III)A, caused by flipping of the N\u2014OH group. That disorder also causes disorder of the nearby ring atoms. Therefore the ring atoms of both the five- and six-membered rings were included in the disorder (but the OAc groups were left out). The geometry of the minor component was restrained to be similar to that of the major one with SAME, SADI and SIMU 0.01 restraints. The refinement of the occupation factors revealed an occupation ratio of 0.802\u2005(7)/0.198\u2005(7) for the two disordered components = 1.2Ueq(C) with the exception of methyl hydrogen atoms, which were placed in their expected positions with HFIX 137 and refined with Uiso(H) = 1.5Ueq(C). For the minor disordered component in compound (II)2 groups (HFIX 13 and 23) and 0.83\u2005\u00c5 for OH groups (HFIX 147), and with Uiso(H) = 1.2Ueq(C) and 1.5Ueq(O). Crystal data, data collection and structure refinement details are summarized in Table\u00a04In compound (II)10.1107/S2056989016018727/zl2684sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989016018727/zl2684Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016018727/zl2684IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989016018727/zl2684IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989016018727/zl2684Isup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016018727/zl2684IIsup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016018727/zl2684IIIsup7.cmlSupporting information file. DOI: 1518715, 1518714, 1518713CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Both tungsten atoms have a slightly distorted octa\u00adhedral coordination.In the title complex two W(CO) 2P:P\u2032]bis\u00ad[penta\u00adcarbonyl\u00adtungsten(0)], [W2(C26H24P2)(CO)10], consists of two W(CO)5 moieties bridged by a bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)ethane (dppe) ligand. The W0 atom has a slightly distorted octa\u00adhedral coordination environment consisting of 5 carbonyl ligands and one P atom from the bridging dppe ligand with the nearest W0 atom 5.625\u2005(5)\u2005\u00c5 away. The complex resides on a center of symmetry.The centrosymmetric title complex, [\u03bc-ethane-1,2-diylbis(di\u00adphenyl\u00adphosphane)-\u03ba Dickson et al. (19892P(CH2)nPPh2 , finding that the predominate product in the reactions of n = 2 and 5 is the bridged complex (OC)5W[\u03bc-Ph2P(CH2)n]PPh2)W(CO)5, whereas when n = 4 it was reported the chelated product is favored (W(CO)4[\u03bc-Ph2P(CH2)4PPh2]. Tan et al. W(CO)5 )5 Fig.\u00a01 consists5W(\u03bc-dppe)W(CO)5 are bridged by a diphosphine approximately along the c axis and the mol\u00adecules themselves are stacked along the a axis. No significant van der Waals-type inter\u00adactions such as C\u2014H\u22ef\u03c0 or \u03c0\u2013\u03c0 contacts between adjacent mol\u00adecules are observed.The two tungsten atoms in each of the mol\u00adecules (OC)5W[\u03bc-Ph2P(CH2)5PPh2]W(CO)5 W(CO)5 2PPh2]Mo(CO)5 5PPh2]Cr(CO)5 5(NH2C6H5) and dppe to produce a golden yellow solution. After two\u2005h, methanol was added to precipitate a yellow solid. The precipitate was collected and washed with methanol (3 x 20\u2005mL). The resulting yellow solid was recrystallized from a 1:5 mixture of di\u00adchloro\u00admethane:methanol at 253\u2005K.All synthesis and crystallization procedures were carried out using standard Schlenk techniques. Di\u00adchloro\u00admethane was added to a mixture of W(CO)Crystal data, data collection, and structure refinement details are summarized in Table\u00a0110.1107/S2056989016013670/vn2114sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016013670/vn2114Isup2.hklStructure factors: contains datablock(s) I. DOI: 1500991CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "O\u2014H\u22efN hydrogen bonds link mol\u00adecules into a supra\u00admolecular chain along 21H28N4O, consists of two unique mol\u00adecules linked by an O\u2014H\u22efN hydrogen bond. The conformation of both C=N bonds is E and the azomethine functional groups lie close to the plane of their associated benzene rings in each of the independent mol\u00adecules. The dihedral angles between the two benzene rings are 83.14\u2005(4) and 75.45\u2005(4)\u00b0. The plane of the one of the N(CH3)2 units is twisted away from the benzene ring by 18.8\u2005(2)\u00b0, indicating loss of conjugation between the lone electron pair and the benzene ring. In the crystal structure, O\u2014H\u22efN hydrogen bonds together with C\u2014H\u22efO hydrogen bonds link neighbouring supra\u00admolecular dimers into a three-dimensional network.The asymmetric unit of the title compound, C Crystal structures of Schiff bases derived from substituted benzaldehydes and 1,3-di\u00adamino\u00adpropan-2-ol have been reported earlier is shown in Fig.\u00a02A and N2 positions can be described by the torsion angles N2A\u2014C5A\u2014C51A\u2014C56A [\u22129.9\u2005(11) in mol\u00adecule A] and N2\u2014C5\u2014C51\u2014C56 [\u221214.9\u2005(11)\u00b0 in mol\u00adecule B]. The two outer aromatic rings (C11\u2013C16 and C51\u2013C56) are inclined to one another by 83.14\u2005(4)\u00b0 in mol\u00adecule A and 75.45\u2005(4)\u00b0 in mol\u00adecule B.The title compound crystallizes with two unique mol\u00adecules in the asymmetric unit. The conformers, labeled et al., 2016bA and 1.271\u2005(8) and 1.269\u2005(8)\u2005\u00c5 in mol\u00adecule B] and the corresponding inter\u00adnal angles at the nitro\u00adgen atom [C1A\u2014N1A\u2014C2A = 117.7\u2005(6) and C5A\u2014N2A\u2014C4A = 117.7\u2005(6) in mol\u00adecule A and C1\u2014N1\u2014C2 = 117.5\u2005(6) and C5\u2014N2\u2014C4 = 117.6\u2005(6) in mol\u00adecule B] also agree with those reported in the literature for similar compounds \u00b0 and C4A\u2014N2A\u2014C5A\u2014C51A = 179.9\u2005(6)\u00b0 in mol\u00adecule A and C2\u2014N1\u2014C1\u2014C11 = 178.6\u2005(6)\u00b0 and C4\u2014N2\u2014C5\u2014C51 = 177.0\u2005(6) in mol\u00adecule B.Bond distances and angles in the benzene rings are not unusual and compare well, both between the two independent mol\u00adecules and with those observed in related systems \u00b0 and C57\u2014N4\u2014C54\u2014C53 = \u22122.9\u2005(11)\u00b0 and with dihedral angles between the NMe2 plane and the benzene ring of 0.57\u2005(2) and 4.60\u2005(2)\u00b0, respectively, whilst in mol\u00adecule A the corresponding torsional angles C17A\u2014N3A\u2014C14A\u2014C13A and C57A\u2014N4A\u2014C54A\u2014C53A are 2.2\u2005(11) and 8.3\u2005(10)\u00b0, respectively. The dihedral angles between the two di\u00admethyl\u00adamino groups (N3A and N4A) and the benzene rings are 5.09\u2005(22) and 18.8\u2005(2)\u00b0 respectively, indicating that the lone electron pair of the N4A atom may not be completely conjugated with the benzene ring (C51A\u2013C56A).The two di\u00admethyl\u00adamino substituents in mol\u00adecule A\u2014H1A and the nitro\u00adgen N2 (Table\u00a01C(5) chains running along the a axis \u2005\u00c5] between O12 Table\u00a01, the twois Fig.\u00a03. The chas Table\u00a01.et al., 2016N,N\u2032-[bis\u00ad]bis\u00ad(4-methyl\u00adbenzene\u00adsulfonamide) -(2-chloro\u00adbenzyl\u00adidene)amino]\u00adpropan-2-ol = 1.5Ueq(O). The remaining H atoms were positioned geometrically and allowed to ride on their parent atoms, with d(C\u2014H) = 0.95\u2005\u00c5 for aromatic and azomethine atoms, d(C\u2014H) = 0.98\u2005\u00c5 for methyl, d(C\u2014H) = 0.99\u2005\u00c5 for methyl\u00adene, d(C\u2014H) = 1.00\u2005\u00c5 for tertiary CH. The Uiso(H) values were constrained to 1.5Ueq(Cmeth\u00adyl) or 1.2Ueq(C) for the remaining H atoms. The structure shows signs of a superstructure. The two mol\u00adecules are related by a translation of 1/2 along the a axis. However, if the structure is refined in a cell with the a axis halved, the displacement parameters of one NMe2 group and some of the C atoms of the phenyl ring to which this group is attached are significantly enlarged I. DOI: 10.1107/S2056989017006429/sj5528Isup2.hklStructure factors: contains datablock(s) I. DOI: 1547455CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III(DMC) acetyl\u00adide complexes that take on a pseudo-octa\u00adhedral symmetry and can be synthesized under weak-base conditions.The crystal structures presented herein consist of two positively charged Co III(DMC) . Chlorido\u00ad(phenyl\u00adethyn\u00adyl)cobalt(III) chloride\u2013aceto\u00adnitrile\u2013methanol (1/1/1), [Co(C8H5)Cl(C12H28N4)]Cl\u00b7CH3CN\u00b7CH3OH, 1, and bis\u00ad(phenyl\u00adethyn\u00adyl)cobalt(III) tri\u00adfluoro\u00admethane\u00adsulfonate\u2013di\u00adchloro\u00admethane (2/1), [Co(C8H5)2(C12H28N4)]2(CF3SO3)2\u00b7CH2Cl2, 2, were prepared under weak-base conditions in satisfactory yields. Single-crystal X-ray diffraction studies revealed that both 1 and 2 adopt a pseudo-octa\u00adhedral symmetry in which the Cl\u2014Co\u2014C angles of 1 and C\u2014Co\u2014C of 2 range from 177.7\u2005(2) to 178.0\u2005(2)\u00b0 and from 177.67\u2005(9) to 179.67\u2005(9)\u00b0, respectively. In both structures, the CoIII metal center is coordinated in the equatorial plane by four N atoms, in which the N\u2014Co\u2014N angles range from 85.6\u2005(3) to 94.4\u2005(3)\u00b0. The structure of 1 features two crystallographically independent mol\u00adecules in its triclinic cell (Z\u2032 = 2), which are related to each other by pseudo-monoclinic symmetry. The crystal investigated was twinned by a symmetry operator of the approximate double-volume C-centered cell , with a refined twin ratio of 0.798\u2005(3) to 0.202\u2005(3). Both methanol solvent mol\u00adecules in 1 are disordered, the major occupancy rates refined to 0.643\u2005(16) and 0.357\u2005(16). Compound 2 also contains two mol\u00adecules in the asymmetric unit, together with two tri\u00adfluoro\u00admethane\u00adsulfonate anions and a disordered di\u00adchloro\u00admethane [occupancy values of 0.545\u2005(12) and 0.455\u2005(12)].Reported in this contribution are the synthesis and crystal structures of new mono- and bis-phenyl\u00adacetyl\u00adides based on Co Inter\u00ade1 crystallizes in PIII(DMC)(C2Ph)(Cl)]+ cation, a chloride counter-ion, and one aceto\u00adnitrile and methanol solvate mol\u00adecule, for a total composition of C20H33ClCoN4\u00b7C2H3N\u00b7CH4O\u00b7Cl. The two unique moieties, labeled A and B, are related by a pseudo-glide plane (see the Supra\u00admolecular features section for a more detailed discussion), and a common atom-naming scheme was used for the contents of the two unique halves of the structure. Both methanol mol\u00adecules are disordered, with a common refined occupancy ratio of 0.643\u2005(16):0.357\u2005(16).Compound 2 crystallizes in P21, Fig.\u00a021, 2 also features two unique cations and anions in its asymmetric unit, but they are not related by any crystallographic pseudo-symmetry. Each complex cation [CoIII(DMC)(C2Ph)2]+ is paired with a triflate anion. The asymmetric unit is completed by a single methyl\u00adene chloride solvate mol\u00adecule, yielding a formula of 2(C28H38CoN4)\u00b72(CF3O3S)\u00b7CH2Cl2. One of the triflate anions as well as the methyl\u00adene chloride mol\u00adecule were refined as disordered, with occupancy rates of 0.503\u2005(22) and 0.545\u2005(12) for the major components.Compound 1 and 2 are similar and 178.0\u2005(2)\u00b0] and C\u2014Co\u2014C [177.67\u2005(9) and 179.67\u2005(9)\u00b0] units are close to normal to the equatorial plane created by the coordinated amines of the macrocyclic ligand, confirming octa\u00adhedral geometries. The C\u2014Co\u2014N and Cl\u2014Co\u2014N angles are all tightly clustered around 90\u00b0. The actual values range from 87.1\u2005(1) to 92.9\u2005(1)\u00b0 \u2013CH2\u2013CH2\u2013N connections to 174.2\u2005(2)\u00b0, with the latter extreme value belonging to one of the Co\u2014C\u2261C units of 1 are as expected for this class of compounds and compare well to values observed by Shores for the cyclam macrocyclic counterpart of 1. \u2005\u00c5 avg.] relative to compound 1. This effect is a result of the stronger \u03c0-donation from phenyl\u00adacetyl\u00adide compared to chloride. The C\u2014C and C\u2261C bond lengths of the phenyl\u00adacetyl\u00adene ligands fall in the expected region for single and triple bonds respectively. The acetyl\u00adides in compound 2 show a slightly cumulenic character with elongated C\u2261C and shortened C\u2014C bond lengths with respect to compound 1, as was also seen by Shores and coworkers and 1.905\u2005(7)\u2005\u00c5] for compound C-centered unit cell with parameters a = 34.721, b = 9.690, c = 15.668\u2005\u00c5, and \u03b2 = 93.41\u00b0. The \u03b1 and \u03b3 angles in the monoclinic cell deviate substanti\u00adally from 90\u00b0, being 88.97 and 89.52\u00b0 when not constrained during data integration. In the crystal structure, the monoclinic pseudo-symmetry manifests itself by the presence of a pseudo b-glide operation along the a-axis of the triclinic cell, Fig.\u00a03A and B, of the surrounding chloride anions and solvate mol\u00adecules and of a second cation. The pseudo-glide symmetry is mostly obeyed by the constituents of the asymmetric unit; the root-mean-square deviation for one overlaid pair of A and B cations is 0.138\u2005\u00c5. For the surrounding solvate mol\u00adecules, for the chloride anions and neighboring cations this is no longer the case. This can especially be seen for a second cation shown in Fig.\u00a04SHELXL) resulted in a twinning ratio of 0.798\u2005(3):0.202\u2005(3), and R1 does increase by 2.6% if twinning is ignored during structure refinement.The structure of the chlorine salt exhibits monoclinic pseudo-symmetry, emulating a double-volume a and c-axes, are shown in Figs. 5Overlays of a larger segment of the lattice, along the 1 of the mono\u00adacetyl\u00adide, the ammonium N\u2014H units of the macrocycle form N\u2014H\u22efN hydrogen bonds with the aceto\u00adnitrile nitro\u00adgen atom, N\u2014H\u22efO hydrogen bonds to the methanol oxygen, and N\u2014H\u22efCl hydrogen bonds to both the inter\u00adstitial chloride anions as well as the cobalt-bound chlorine. The chloride anions are also acceptors for O\u2014H\u22efCl hydrogen bonds originating from the disordered methanol mol\u00adecules and for a series of weaker C\u2014H\u22efCl hydrogen bonds from macrocyclic carbon atoms. The type and number of hydrogen bonds is essentially the same between the two halves of the structure related by pseudo-symmetry, but the exact metrics and numbers are slightly modulated. The N\u2014H\u22efN, N\u2014H\u22efO, and N\u2014H\u22efCl hydrogen bonds, when combined, connect the cations, anions and solvate mol\u00adecules into ribbons that extend infinitely along the b-axis and are perpendicular to the a-axis, and exactly one unit cell thick in the a- and c-axis directions , and their strength varies substanti\u00adally between the ion pairs. The first of the two cations, involving nitro\u00adgen atoms N1 through N4, features each two N\u2014H\u22efO and N\u2014H\u22efF hydrogen bonds (not counting duplicates from triflate disorder), Fig.\u00a09In the triflate salt ns Fig.\u00a08. The twoIII(DMC)Cl2]Cl was synthesized according to literature procedures (C2Ph)Cl]Cl (1). [CoIII(DMC)Cl2]Cl was dissolved in 40\u2005mL of methanol. Phenyl\u00adacetyl\u00adene and Et3N were added and the solution was refluxed overnight. Solvent was removed via rotary evaporation, and the solid was loaded onto a silica gel plug and eluted with CH3OH/EtOAc as a red fraction. The desired product was recrystallized from ether\u2013methanol to afford 170\u2005mg of a coral solid (73% based on [CoIII(DMC)Cl2]Cl). Single crystals were grown from slow diffusion of ether into a methanol solution of 1.Preparation of [CoIII(DMC)(C2Ph)Cl]Cl (1). ESI\u2013MS: [M]+, 423.0. 1H NMR : 7.55\u20137.41 , 7.37\u20137.25 , 7.25\u20137.15 , 5.36 , 4.23 , 3.21\u20132.46 , 1.93\u20131.84 , 1.53\u20131.48 , 1.30 . Visible spectra, \u03bbmax : 256 , 493\u2005(101); IR (cm\u22121): C\u2261C: 2122 (m).Data for [CoIII(DMC)(C2Ph)2]OTf (2). Compound 1 and AgOTf were dissolved in 50\u2005mL of CH3CN and refluxed for 48\u2005h. The precipitate that formed was filtered out, and 3.1\u2005mL (22\u2005mmol) of Et3N and 0.20\u2005mL (1.8\u2005mmol) of phenyl\u00adacetyl\u00adene were added and the solution was refluxed for 48\u2005h. The solution was purified over a silica gel plug and the product eluted with CH3OH/EtOAc . A pale-yellow fraction was collected and recrystallized from ether\u2013methanol to afford 102\u2005mg of a yellow solid (47% based on 1). Single crystals were grown from slow diffusion of n-hexa\u00adnes into a CH3OH/CH2Cl\u00ad2 solution of 2.Preparation of [CoIII(DMC)(C2Ph)2]OTf (2). ESI\u2013MS: [M]+, 489.0. 1H NMR : 7.58\u20137.42 , 7.36\u20137.24 , 7.22\u20137.13 , 4.90 , 3.84 , 3.30\u20133.01 , 2.81\u20132.78 , 2.68\u20132.63 , 2.50\u20132.43 , 1.83 , 1.38 , 1.27 . Visible spectra, \u03bbmax : 271 , 464 (64.5); IR (cm\u22121): C\u2261C: 2102 (m).Data for direction in reciprocal space . Application of the twin matrix 1 0 0, 0 The structure of compound 1, each methanol group was refined with two-component disorder with a shared occupancy ratio for the two sites. The C\u2014O bond lengths were restrained to 1.427\u2005(20)\u2005\u00c5. Each minor occupancy component was restrained to be similar the respective major occupancy component . The Uij components for atoms within 2.0\u2005\u00c5 were restrained to be similar . The alcohol hydrogen atom to neighboring chloride distances were restrained based on hydrogen-bonding considerations. Subject to these conditions, the occupancy rates refined to 0.643\u2005(16) and 0.357\u2005(16).In the structure of compound 2, the S1 triflate anion was refined with two-component disorder. Each moiety was restrained to have a similar geometry as the S2 triflate anion . The Uij components for disordered atoms within 2.0\u2005\u00c5 were restrained to be similar . Subject to these conditions, the occupancy factors refined to 0.503\u2005(22) and 0.497\u2005(22). The di\u00adchloro\u00admethane mol\u00adecule was refined with two-component disorder. The minor occupancy component was restrained to have a similar geometry as the major occupancy component . The Uij components for atoms within 2.0\u2005\u00c5 were restrained to be similar . Subject to these conditions, the occupancy factors refined to 0.545\u2005(12) and 0.455\u2005(12).In the structure of compound 10.1107/S2056989018003997/vm2209sup1.cifCrystal structure: contains datablock(s) 1, 2, global. DOI: 10.1107/S2056989018003997/vm22091sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989018003997/vm22092sup3.hklStructure factors: contains datablock(s) 2. DOI: 1828222, 1828221CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "R,R,SFc,SFc)-[Fe2(C5H5)2(C38H36BrNP2)]\u00b7Br\u00b7CH2Cl2 has been determined by X-ray single-crystal diffraction.The absolute structure of -[Fe2(C5H5)2(C38H35NP2)] (1), single crystals of the di\u00adchloro\u00admethane monosolvate of the Br\u2212 salt of the protonated ligand 1H+ were obtained serendipitously, i.e. [Fe2(C5H5)2(C38H36NP2)]Br\u00b7CH2Cl2. The crystal structure of 1H\u00b7Br\u00b7CH2Cl2 was determined by single-crystal X-ray diffraction. The mean bond lengths in the ferrocene units are Fe\u2014C = 2.049\u2005(3)\u2005\u00c5 and C\u2014C = 1.422\u2005(4)\u2005\u00c5 within the cyclo\u00adpenta\u00addienyl rings. The mean C\u2014N bond length is 1.523\u2005(4)\u2005\u00c5. The inter\u00adplanar angle between the two connected cyclo\u00adpenta\u00addienyl rings is 49.2\u2005(2)\u00b0. One ferrocene moiety adopts a staggered conformation, whereas the other is between staggered and eclipsed. The Br\u2212 ions and the CH2Cl2 mol\u00adecules are located in channels extending along <100>. One ammonium H atom forms a hydrogen bond with the Br\u2212 ion [H\u22efBr = 2.32\u2005(4)\u2005\u00c5 and C\u2014H\u22efBr = 172\u2005(3)\u00b0]. The second ammonium H atom is not involved in hydrogen bonding.During the synthesis of an FeBr Ammonium H atoms were found in difference Fourier maps and were refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016020417/pk2595sup1.cifCrystal structure: contains datablock(s) I, general. DOI: 10.1107/S2056989016020417/pk2595Isup2.hklStructure factors: contains datablock(s) I. DOI: 1524191CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The hig\u201d should be replaced with \u201c50 values of H2O2 were found for cell pretreated with 50\u2009\u03bcg/mL BJLN (89.95\u2009\u03bcM) and 100\u2009\u03bcg/mL MR219 (143.90\u2009\u03bcM) extracts when compared to negative control (271.00\u2009\u03bcM) Significant decrements in the IC1.00\u2009\u03bcM) . The hig.\u201dIn addition, in the \u201cResults\u201d (Section 3.3), the text reading \u201cAn incorrect version of \u03bcg/mL) (grey dotted line) was incorrectly plotted. The correct version of the figure is as shown below with the corrected fourth datum point for MR219 (50\u2009\u03bcg/mL) (grey dotted line).Accordingly,"} +{"text": "In the crystal, the ASP anions are linked 14H16NO+\u00b7C4H6NO4\u2212\u00b7H2O, crystallizes as a monohydrate. The 1,2-di\u00adphenyl\u00adethyl group in the cation has a cis conformation, and the aspartic acid anion is in the zwitterionic form. In the crystal, the ASP anions are linked via N\u2014H\u22efO hydrogen bonds to form a 21 helix along the b-axis direction. The helices are linked by the ADE cations via O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming layers parallel to the bc plane. There are channels in the layers that are occupied by water mol\u00adecules, which link to both the anions and cations via Owater\u2014H\u22efO and N\u2014H\u22efOwater hydrogen bonds. There are also C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions present within the layers.The title diastereomeric salt, formed between 2-amino-1,2-di\u00adphenyl\u00adethanol (ADE) and aspartic acid (ASP), C The optical separation of ASP with cis-ADE was introduced without chemical modification. The crystal structure of the title mol\u00adecular salt, formed between l-(S)-ASP and -cis-ADE, is reported herein.The production of chiral compounds has great importance in the pharmaceutical industry, and diastereomeric salt separation is still widely applied in the process. A synthetic optical resolving agent, chiral 2-amino-1,2-di\u00adphenyl\u00adethanol (ADE) is 48.71\u2005(9)\u00b0 and the torsion angle O1A\u2014C1A\u2014C2A\u2014N1A is \u221265.0\u2005(2)\u00b0. The hy\u00addroxy group adopts a gauche conformation [O1A\u2014C1A\u2014C2A\u2014C9A = 60.1\u2005(2)\u00b0] with respect to phenyl ring B. Thus, the tweezer-like motif is twisted with respect to the phenyl groups. This arrangement is similar to that found in racemic cis-ADE -ASP crystallizes as a deprotonated zwitterion. The succinate group adopts a cis conformation [C1B\u2014C2B\u2014C3B\u2014C4B = \u221253.0\u2005(2)\u00b0], which is the motif commonly found in l-ASP salts; for example l-His\u00b7l-ASP monohydrate and 1.4\u2005(3)\u00b0, respectively.S)-ASP anions correlated with crystallographic symmetry are linked via N1B\u2014H1B3\u22efO4Bii [2.868\u2005(2)\u2005\u00c5] hydrogen bonds into C(6) chains to form a right-handed 12-helix along the b-axis direction -cis-ADE cations via N\u2014H\u22efO hydrogen bonds [N1A\u2014H1A2\u22efO1B = 2.862\u2005(2)\u2005\u00c5 and N1A\u2014H1A1\u22efO4Biii = 2.742\u2005(3)\u2005\u00c5] and O\u2014H\u22efO hydrogen bonds [O1A\u2014H1O1\u22efO2Bi = 2.752\u2005(2)\u2005\u00c5], forming layers parallel to the bc plane \u2005\u00c5 and O1C\u2014H1OA\u22efO1Biv = 2.840\u2005(2)\u2005\u00c5] and N\u2014H\u22efOwater hydrogen bonds [N1B\u2014H1B2\u22efO1C = 2.938\u2005(2)\u2005\u00c5 and N1A\u2014H1A3\u22efO1C = 2.926\u2005(3)\u2005\u00c5], shown in Fig.\u00a04a-axis direction.In the crystal, the -cis-2-Amino-1,2-di\u00adphenyl\u00adethanol (ADE) and aspartic acid (ASP) were purchased from Sigma\u2013Aldrich Co. Ltd. The title mol\u00adecular salt was obtained from an aqueous ethanol solution of racemic-ASP and -cis-ADE in a 2:1 molar ratio, heated to 333\u2005K under stirring. On slow cooling to ambient temperature and slow evaporation of the solvent, colourless rod-shaped crystals were obtained. = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017015729/su5398sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989017015729/su5398Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017015729/su5398Isup3.cmlSupporting information file. DOI: 1582706CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecule has an almost planar geometry (r.m.s. deviation = 0.012\u2005\u00c5), and its mol\u00adecular structure is stabilized by an intra\u00admolecular secondary Se\u22efO inter\u00adaction of 2.353\u2005(3)\u2005\u00c5, closing a four-membered N\u2014C\u2014Se\u22efO ring. The title compound represents the first monomeric organoselenenyl chloride stabilized intra\u00admolecularly by an inter\u00adaction of this type. The non-valent attractive Se\u22efO inter\u00adaction results in a substantial distortion of the geometry of the ipso-carbon atom. The endo-cyclic N\u2014C\u2014Se [102.1\u2005(3)\u00b0] and exo-cyclic C\u2014C\u2014Se [136.9\u2005(3)\u00b0] bond angles deviate significantly from the ideal value of 120\u00b0 for an sp2-hybridized carbon atom, the former bond angle being much smaller than the latter. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, forming zigzag chains propagating along [010]. The chains, which stack along the a-axis direction, are linked by offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.960\u2005(3)\u2005\u00c5], forming corrugated sheets parallel to the ab plane.The title compound, C RSeX play an important role in modern organic synthesis and are used as reagents for the functionalization of many classes of compounds, including organoselenium compounds with a broad spectrum of biologi\u00adcal activities \u00b0] and exo-cyclic C3\u2014C2\u2014Se1 [136.9\u2005(3)\u00b0] bond angles deviate significantly from the ideal value of 120\u00b0 for an sp2-hybridized carbon atom, the former angle being much smaller than the latter. The title compound represents the first monomeric organoselenenyl chloride stabilized intra\u00admolecularly by an inter\u00adaction of this type. Previously, the analogous stabilization of monomeric organoselenenyl chlorides by intra\u00admolecular secondary Se\u22efS \u2005\u00c5, Cg is the centroid of the N1/C2\u2013C6 ring, inter\u00adplanar distances = 3.590\u2005(2)\u2005\u00c5, slippages = 1.671\u2005\u00c5, symmetry codes: (i) x\u00a0\u2212\u00a01, y, z; (ii) x\u00a0+\u00a01, y, z].In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds Table\u00a01, forminget al. of the title compound. Colourless prismatic crystals of the title compound were obtained after recrystallization of the crude product from di\u00adchloro\u00admethane (m.p. 433\u2013435\u2005K). IR , \u03bd 1617, 1462, 1423, 1254, 1151, 836, 748, 621. 1H NMR : \u03b4 = 8.28 ; 7.52 ; 7.43 ; 7.30 . Analysis calculated for C5H4ClNOSe: C 24.81; H 1.93; N 6.72. Found: 24.43; H 1.83; N 6.65.The synthesis of the title compound is illustrated in Fig.\u00a03 2010ab,c \u25b8. A Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016018946/su5337sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989016018946/su5337Isup2.hklStructure factors: contains datablock(s) I. DOI: 1519449CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The 1:1 co-crystal comprising two fused-ring mol\u00adecules features significant hydrogen bonding between the 1,8-di\u00adhydroxy\u00adanthra\u00adquinone coformers with the main links between the resulting dimeric aggregates and the bromo\u00adnaphtho\u00adquinone coformer being of the type C\u2014H\u22efO. 10H5BrO2\u00b7C14H8O4 , features one mol\u00adecule of each coformer. The 2-bromo\u00adnaphtho\u00adquinone mol\u00adecule is almost planar . The 1,8-di\u00adhydroxy\u00adanthra\u00adquinone mol\u00adecule is planar (r.m.s. deviation for the 18 non-H atoms is 0.022\u2005\u00c5) and features two intra\u00admolecular hy\u00addroxy-O\u2014H\u22efO(carbon\u00adyl) hydrogen bonds. Dimeric aggregates of 1,8-di\u00adhydroxy\u00adanthra\u00adquinone mol\u00adecules assemble through weak inter\u00admolecular hy\u00addroxy-O\u2014H\u22efO(carbon\u00adyl) hydrogen bonds. The mol\u00adecular packing comprises stacks of mol\u00adecules of 2-bromo\u00adnaphtho\u00adquinone and dimeric assembles of 1,8-di\u00adhydroxy\u00adanthra\u00adquinone with the shortest \u03c0\u2013\u03c0 contact within a stack of 3.5760\u2005(9)\u2005\u00c5 occurring between the different rings of 2-bromo\u00adnaphtho\u00adquinone mol\u00adecules. The analysis of the Hirshfeld surface reveals the importance of the inter\u00adactions just indicated but, also the contribution of additional C\u2014H\u22efO contacts as well as C=O\u22ef\u03c0 inter\u00adactions to the mol\u00adecular packing.The asymmetric unit of the title co-crystal, C The six carbon atoms comprising the cyclo\u00adhexa-2,5-diene-1,4-dione ring of the naphtho\u00adquinone mol\u00adecule are not strictly planar with the r.m.s. deviation being 0.030\u2005\u00c5; the maximum deviations are 0.025\u2005(1) and \u22120.031\u2005(2)\u2005\u00c5 for the C4a and C4 atoms, respectively. The appended Br1, O1 and O4 atoms lie, respectively, 0.077\u2005(1), 0.078\u2005(1) and \u22120.117\u2005(1)\u2005\u00c5 out of the plane with the Br1 atom lying to one side of the ring and the carbonyl-O atoms to the other. Overall, the r.m.s. deviation for the best plane defined by the 13 non-H atoms comprising the naphtho\u00adquinone mol\u00adecule is 0.060\u2005\u00c5, with the maximum deviations being 0.093\u2005(1)\u2005\u00c5 for atom Br1 and \u22120.099\u2005(1)\u2005\u00c5 for the O4 atom, again with these atoms lying to opposite sides of the plane. With respect to the anthra\u00adquinone mol\u00adecule, the r.m.s. deviation for the 18 non-H atoms is 0.022\u2005\u00c5 with the maximum deviations being 0.039\u2005(2)\u2005\u00c5 for C(13) and 0.026\u2005(1)\u2005\u00c5 for the C19 and C23 atoms. As seen from Fig.\u00a01b, the hy\u00addroxy-H atoms are orientated to be proximate to the centrally located carbonyl-O atom to form intra\u00admolecular hy\u00addroxy-O\u2014H\u22efO(carbon\u00adyl) hydrogen-bonds, Table\u00a01The mol\u00adecular structures of the constituents of (I)a, is connected by a centrosymmetric planar, eight-membered {\u22efHO\u22efO\u22efH}2 synthon which incorporates two transannular hy\u00addroxy-O\u2014H\u22efO(carbon\u00adyl) hydrogen bonds. The dimeric aggregates stack along the b axis being surrounded by two columns of similar dimeric aggregates and six columns comprising naphtho\u00adquinone mol\u00adecules, Fig.\u00a02b. Connections between columns, leading to a three-dimensional architecture, are of the type sp2-C\u2014H\u22efO(carbon\u00adyl) and involve all the remaining carbonyl-O atoms with the O atom of the naphtho\u00adquinone-C4=O4 moiety forming two such contacts, Table\u00a01i.e. between the and rings with the inter-centroid separation being 3.5760\u2005(9)\u2005\u00c5 and the angle of inclination being 1.64\u2005(7)\u00b0 for symmetry operation x, \u22121\u00a0+\u00a0y, z. The closest comparable inter\u00adaction within the stack of anthra\u00adquinone mol\u00adecules is 4.1013\u2005(9)\u2005\u00c5, i.e. between (C15\u2013C21) and (C19\u2013C24) rings; angle of inclination = 0.65\u2005(7)\u00b0 for symmetry operation: x, \u22121\u00a0+\u00a0y, z.In addition to the intra\u00admolecular hy\u00addroxy-O\u2014H\u22efO(carbon\u00adyl) hydrogen-bonds in the anthra\u00adquinone mol\u00adecule, both hy\u00addroxy-H atoms from weaker inter\u00admolecular hydrogen-bonds with a centrosymmetrically related mol\u00adecule indicating each hy\u00addroxy-H atom is bifurcated, Table\u00a01et al., 2016et al., 2016dnorm, Figs. 3The Hirshfeld surface analysis of title 1:1 co-crystal, (I)dnorm in Fig.\u00a03a. On the Hirshfeld surface mapped over the calculated electrostatic potential, the respective donors and acceptors appear as the blue and red regions in Fig.\u00a05a. The presence of faint-red spots near carbon atoms C11, C19, Fig.\u00a03a, and near the atoms C15 and C20, Fig.\u00a03b, also indicate the links between mol\u00adecules through short inter-atomic C\u22efC contacts, Table\u00a02a. Links between the coformers involving their carbonyl-C4=O4 and C20=O20 groups through short inter\u00adatomic C\u22efO/O\u22efC contacts, Table\u00a02b and 4b. This is also illustrated by the black dashed lines on the Hirshfeld surface mapped over electrostatic potential in Fig.\u00a06b. The donors and acceptors of inter\u00admolecular C\u2014H\u22efO(carbon\u00adyl) inter\u00adactions can be viewed as bright-red spots having labels \u20183\u2019\u2013\u20185\u2019 in Figs. 3b and 4a, labelled with \u20186\u2019. The immediate environments about reference anthra\u00adquinone and naphtho\u00adquinone mol\u00adecules within shape-index-mapped Hirshfeld surfaces highlighting inter\u00admolecular O\u2014H\u22efO, C\u2014H\u22efO, \u03c0\u2013\u03c0 stacking and C\u2014O\u22ef\u03c0 inter\u00adactions influential on the packing are illustrated in Figs. 7The donors and acceptors of inter\u00admolecular hy\u00addroxy-O\u2014H\u22efO(carbon\u00adyl) hydrogen-bonds between anthra\u00adquinone mol\u00adecules are viewed as bright-red spots labelled with \u20181\u2019 and \u20182\u2019 on the Hirshfeld surfaces mapped over a. The plots delineated into H\u22efH, O\u22efH/H\u22efO, C\u22efH/H\u22efC, C\u22efC and C\u22efO/O\u22efC contacts atoms are viewed as a pair of short spikes at de\u00a0+\u00a0di\u00a0\u223c\u00a02.4\u2005\u00c5 in the donor and acceptor regions of their respective plots in Fig.\u00a09c. The points corresponding to anthra\u00adquinone-C15\u2014H15\u22efO4(carbon\u00adyl) inter\u00adactions and other short inter\u00adatomic O\u22efH contacts, Table\u00a02The donors and acceptors of the naphtho\u00adquinone-H3 and anthra\u00adquinone-O20(carbon\u00adyl) atoms are viewed as a thin, long spike at de\u00a0+\u00a0di\u00a0<\u00a02.9\u2005\u00c5, i.e. less than sum of their van der Waals radii, in the fingerprint plot delineated into C\u22efH/H\u22efC contacts for anthra\u00adquinone, Fig.\u00a09d, are indicative of short inter\u00adatomic C\u22efH contacts, Table\u00a02e, having high density at around de\u00a0=\u00a0di\u00a0\u223c\u00a01.8\u2005\u00c5. The parabolic distribution of points with the peak at de\u00a0=\u00a0di\u00a0\u223c\u00a01.6\u2005\u00c5 in the plot for the anthra\u00adquinone coformer, Fig.\u00a09e, indicates links between these mol\u00adecules through short inter\u00adatomic C\u22efC contacts along the b axis. The presence of C\u22efC contacts in (I)A pair of short peaks at f, results from short, inter-atomic C\u22efO/O\u22efC contacts whereas the 11.9% contribution from C\u22efO/O\u22efC contacts for the anthra\u00adquinone mol\u00adecule has a contribution from C=O\u22ef\u03c0 inter\u00adactions involving carbony-O19 and -O20 atoms and and rings, Table\u00a04f.The 3.9% contribution from C\u22efO/O\u22efC contacts to the Hirshfeld surface for the naphtho\u00adquinone mol\u00adecule, Fig.\u00a09Although the naphtho\u00adquinone-bromide substituent makes a notable contribution to the Hirshfeld surface, Table\u00a03et al., 2016cf. 0.060\u2005\u00c5 in (I)et al., 2008i.e. a 3:1 co-crystal with acetic acid 2Cl2/hexane (1:1 v/v) and a single, yellow fraction was collected. After evaporation of the solvent under reduced pressure, a yellow solid was obtained. This was recrystallized from ethyl acetate solution to give small orange\u2013red crystals with yields of 78\u201385% based upon the qu\u00adantity of 1,8-di\u00adhydroxy\u00adanthra\u00adquinone initially used. Notably, the substrates 2-bromo\u00adnaphtho\u00adquinone and 1,8-di\u00adhydroxy\u00adanthra\u00adquinone could not be chromatographically distinguished as they ran with equivalent Rf\u2019s in a wide range of solvents and solvent mixtures. NMR spectra (1H and 13C) were consistent with a one to one mixture of the same components as there was no deviation of chemical shifts in comparison to the spectra of the individual components. A sample of the co-crystal material had a well defined melting point of 413\u2013414\u2005K, which is inter\u00admediate between the melting points of the pure components 2-bromo\u00adnaphtho\u00adquinone, 405\u2013406\u2005K Uiso(H) set to 1.2Ueq(C). The O-bound H atoms were located from a difference map but refined with O\u2014H = 0.84\u00b10.01\u2005\u00c5 and Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989017005667/wm5383sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017005667/wm5383Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017005667/wm5383Isup3.cmlSupporting information file. DOI: 1543933CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I atom is surrounded by three \u03bc3-iodide anions and one S atom, while the other is coordinated by three \u03bc3-iodide ions an O atom. In the crystal, there are inter\u00admolecular C\u2014H\u22efI hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions between ligands. The packing generates a two-dimensional brick-wall structure.The structure of the title copper(I) coordination polymer is reported. One Cu H)-one in aceto\u00adnitrile/di\u00adchloro\u00admethane results in a crystalline coordination polymer, namely poly[bis\u00ad{\u03bc2-1-[2-(cyclo\u00adhexyl\u00adsulfan\u00adyl)eth\u00adyl]pyridin-2(1H)-one}tetra-\u03bc3-iodido\u00adtetra\u00adcopper(I)], [Cu4I4L2]n. The asymmetric unit comprises two ligand mol\u00adecules, four copper(I) ions and four iodide ions. Inter\u00adestingly, the O atoms are bound to the soft copper(I) ions. The stair-step clusters of Cu and I atoms in the asymmetric unit are linked repeatedly, giving rise to infinite chains along [100]. Neighbouring infinite chains are linked through the L mol\u00adecules, forming a two-dimensional brick-wall structure. These two-dimensional networks are stacked alternately along [001]. Additionally, there are inter\u00admolecular C\u2014H\u22efI hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions between the ligands.The reaction of copper(I) iodide with 1-[2-(cyclo\u00adhexyl\u00adsulfan\u00adyl)eth\u00adyl]pyridin-2(1 CuI\u2014O bonds have been reported previously in copper(I) coordination polymers with phosphine ligands complexes have been studied continuously over several decades because of their potential applications as sensors, catalysts, and gas storage materials ions, four \u03bc3-iodide ions, and two L ligands as shown in Fig.\u00a01A (identified by S1) and LB (identified by S2), the pyridyl and cyclo\u00adhexyl rings are in anti and gauche conformations with torsion angles of \u2212154.7\u2005(6)\u00b0 [C6\u2014S1\u2014C7\u2014C8] and 62.3\u2005(7)\u00b0 [C19\u2014S2\u2014C20\u2014C21], respectively. All of the CuI atoms (Cu1\u2013Cu4) have distorted tetra\u00adhedral coordination geometries. The Cu1 and Cu2 atoms are bound by three \u03bc3-iodide anions and one S atom, while Cu3 and Cu4 are coordinated by three \u03bc3-iodide ions and one O atom. The ranges of inter\u00adatomic distances in the title compound are 2.7082\u2005(15)\u20132.7444\u2005(14)\u2005\u00c5, 2.297\u2005(2)\u20132.314\u2005(2)\u2005\u00c5, 2.6210\u2005(12)\u20132.7230\u2005(12)\u2005\u00c5, and 2.071\u2005(6)\u20132.087\u2005(6)\u2005\u00c5 for Cu\u2014Cu, Cu\u2014S, Cu\u2014I, and Cu\u2014O, respectively as shown in Fig.\u00a02A\u22efI4ii, C12\u2014H12\u22efI1iii and C21\u2014H21B\u22efI3iv hydrogen bonds between ligands. Red dashed lines display inter\u00admolecular C5\u2014H5A\u22efCg1v [H\u22efCg1=3.00\u2005\u00c5] inter\u00adactions between the ligands complexes of N/S mixed donor atom ligands have been reported (Jeon Synthesis of 1-[2-(cyclo\u00adhexyl\u00adsulfan\u00adyl)eth\u00adyl]pyridin-2(1H)-one (L)2SO4, and evaporated to give a crude oil. Column chromatography , Rf = 0.28) : 7.28 , 6.52 , 6.11 , 4.01 , 2.85 , 2.51 , 2.00\u20131.13 ; 13C NMR : 161.33, 140.03, 139.52, 119.40, 104.86, 49.21, 42.48, 33.15 27.71, 25.44, 25.29.Thionyl chloride was added dropwise to 2-(cyclo\u00adhexyl\u00adthio)\u00adethanol in chloro\u00adform. The mixture was stirred under reflux for 1\u2005h then cooled to 253\u2005K. Chloro\u00adform was removed, yielding crude 2-chloro\u00adethyl\u00adcyclo\u00adhexyl\u00adsulfide. 2-Hy\u00addroxy\u00adpyridine and potassium hydroxide were dissolved in 10\u2005ml of tetra\u00adhydro\u00adfuran and 5\u2005ml of water, and then the solution was added dropwise to the crude chloride. The solution was refluxed for 24\u2005h and cooled. The crude product was extracted by di\u00adchloro\u00admethane. The di\u00adchloro\u00admethane layer was dried with anhydrous Na4I4Preparation of [CuL2]nL was allowed to mix with an aceto\u00adnitrile (5\u2005ml) solution of CuI . The colourless precipitate was filtered and washed with a diethyl ether/aceto\u00adnitrile (5/1 v/v) solution. Single crystals suitable for X-ray analysis were obtained by slow evaporation of di\u00adchloro\u00admethane from the reaction mixture.A di\u00adchloro\u00admethane (5\u2005ml) solution of Uiso(H) = 1.2Ueq(C) for aromatic C\u2014H groups, C\u2014H = 0.99\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for CH2 groups, and C\u2014H = 1.00\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for Csp3\u2014H groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017015377/sj5538sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989017015377/sj5538Isup2.hklStructure factors: contains datablock(s) I. DOI: 1581394CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound was isolated as a by-product from a reaction incorporating 2,4,6-tri\u00admethyl\u00adbenzoic acid under basic conditions. 20H22O3, was formed in the reaction between 2,4,6-tri\u00admethyl\u00adbenzoic acid and N,N-diiso\u00adpropyl\u00adethyl\u00adamine in the presence of 1,3-di\u00adchloro-1,3-bis\u00ad(di\u00admethyl\u00adamino)\u00adpropenium hydrogen dichloride, and was recrystallized from diethyl ether solution. It is the first exclusively alkyl-substituted benzoic anhydride to have been structurally characterized. The asymmetric unit consists of a half mol\u00adecule, the other half of which is generated by twofold rotation symmetry; the dihedral angle between the symmetry-related aromatic rings is 54.97\u2005(3)\u00b0. The geometric parameters of the aromatic ring are typical of those for 2,4,6-tri\u00admethyl\u00adphenyl substituted groups. The C=O and C\u2014O bond lengths are 1.1934\u2005(12) and 1.3958\u2005(11)\u2005\u00c5, respectively, and the angle between these three atoms (O=C\u2014O) is 121.24\u2005(9)\u00b0. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions. The packing features wavy chains that extend parallel to [001].The title compound, C The C1\u2014C2 distance is normal for an sp2\u2013sp2 bond, with a length of 1.4873\u2005(13)\u2005\u00c5. The second half of the mol\u00adecule, which is generated by rotation about the twofold axis passing through O1 , forms a dihedral angle of 54.97\u2005(3)\u00b0 between the equivalent aromatic rings. If the planes of the two overlapping CO2 groups are chosen instead, the dihedral angle becomes 59.30\u2005(11)\u00b0. The C\u2014C bonds in the aromatic ring are not all statistically equivalent. Unsurprisingly, the longest C\u2014C bonds in the ring are adjacent to the electron-withdrawing anhydride group, C2\u2014C3 [1.4032\u2005(13)\u2005\u00c5] and C2\u2014C7 [1.4059\u2005(13)\u2005\u00c5]. The remaining C\u2014C bonds are statistically equivalent, averaging 1.3942\u2005(8)\u2005\u00c5. All of the C\u2014CH3 bond lengths are statistically equivalent with an average length of 1.5102\u2005(8)\u2005\u00c5.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01a axis, forms wavy chains that run parallel to the c-axis direction and H6 (2.39\u2005\u00c5 to H10A), and the designated methyl H atoms, which close five-membered rings in the mol\u00adecule.There are short intra\u00admolecular contacts between the aromatic H atoms H4 were added to a chloroform solution (25\u2005ml) of 1,3-di\u00adchloro-1,3-bis\u00ad(di\u00admethyl\u00adamino)\u00adpropenium hydrogen dichloride , which had been prepared following the known literature method and then isolated via vacuum filtration. The off-white solid was further purified by recrystallization through slow evaporation of a saturated diethyl ether solution. After 3\u2005h, clear and colourless thin plate-like crystals were obtained . Elemental analysis, calculated for C20H22O3 (%): C\u00a077.39, H 7.14, N 0.00; found (%): C 77.26, H 7.13, N 0.01. The 1H and 13C{1H} NMR and IR spectroscopic data for the title compound are identical to those previously reported = 1.2 or 1.5Ueq(C).Crystal data, data collection, and structure refinement details are summarized in Table\u00a0210.1107/S2056989017014670/hb7710sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017014670/hb7710Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017014670/hb7710Isup3.cmlSupporting information file. DOI: 1579203CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atom in the title compound is seven-coordinate, being bound to two pyridine N atoms, one amine N atom and four O atoms to form a distorted penta\u00adgonal\u2013bipyramidal environment. Each dipyridyl-type ligand links the CoII atoms into polymeric zigzag chains, which are connected via inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions and N/C\u2014H\u22efO hydrogen bonds.The Co 3)2L]n, L = N-(pyridine-2-ylmeth\u00adyl)pyridine-3-amine (C11H11N3), contains one CoII centre, two nitrate anions and one L ligand in which the Cpy\u2014C\u2014N\u2014Cpy moiety adopts a trans conformation with a torsion angle of \u2212173.1\u2005(3)\u2005\u00c5. The coordination geometry of the CoII atom is a distorted penta\u00adgonal bipyramid. One amine N atom from the L ligand and four O atoms from two \u03b72-nitrato ligands form the basal plane and two pyridyl N atoms from two symmetry-related L ligands occupy the apical positions [N\u2014Co\u2014N = 171.86\u2005(11)\u00b0]. The displacement of the central CoII atom from the basal plane (r.m.s. deviation = 0.085\u2005\u00c5) is 0.1491\u2005(12)\u2005\u00c5. Each bidentate nitrate group is bonded asymmetrically to the cobalt atom in an chelating fashion. The CoII ions are linked by the L ligands to form a zigzag chain propagating along the c-axis direction. Within the zigzag chain, C\u2014H\u22efO hydrogen bonds between the ligands and the nitrate anions are observed. Adjacent zigzag chains are connected via inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid-to-centroid distance = 3.844\u2005(2)\u2005\u00c5] between the pyridine rings together with N/C\u2014H\u22efO hydrogen bonds.The asymmetric unit of the title compound, [Co(NO Herein, we report its crystal structure, which is the first example of a CoII complex with an N-(pyridine-2-ylmeth\u00adyl)pyridine-3-amine ligand.Over the past few decades, the continuous efforts have been devoted to the design and development of metal\u2013organic frameworks (MOFs) obtained by linking transition metal centers with several organic bridging ligands. In particular, rigid or flexible dipyridyl-type ligands have been widely used to construct MOFs with attractive structures and potential applications in materials chemistry \u00b0; symmetry code: (i) x, \u2212y\u00a0+\u00a0z\u00a0\u2212\u00a0II atom is displaced by 0.1491\u2005(12)\u2005\u00c5 from the basal plane (r.m.s. deviation = 0.085\u2005\u00c5). The Co\u2014N distances in apical positions are slightly shorter than that of the basal [Co1\u2014N2 = 2.191\u2005(3)\u2005\u00c5]. The largest deviations from the NO4 basal plane around the cobalt center involve the angles O2\u2014Co1\u2014O3 [55.81\u2005(11)\u00b0] and N2\u2014Co1\u2014O5 [84.19\u2005(9)\u00b0]. This distortion may reflect the narrow bite angles of the bidentate nitrate ions.The asymmetric unit of the title compound comprises one Co\u00a0] Fig.\u00a01. The cenL ligand adopts a stretched trans conformation with the C5\u2014C6\u2014N2\u2014C7 torsion angle being \u2212173.1\u2005(3)\u2005\u00c5. The terminal pyridine rings of the L ligand are nearly perpendic\u00adular to each other, with the dihedral angle between their mean planes being 76.74\u2005(12)\u00b0. Each bidentate nitrate group is bonded asymmetrically to the cobalt atom . Each L ligand is bridged by the CoII ions, forming \u2013(Co-L)n\u2013 zigzag chains propagating along the c-axis direction \u2005\u00c5; Cg1 is the centroid of the N1/C1\u2013C5 ring; symmetry code: (ii) \u2212x, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01] between the pyridine rings and C\u2014H\u22efO hydrogen bonds between pyridyl H atoms and nitrate O atoms are HgII complexes and seven of them are AgI complexes. The remaining one is a ZnII complex (DUVPER). There are no metal complexes that are similar to the structure of the CoII complex described above. Therefore, the title compound is the first example of a CoII complex with an L ligand.A search of the Cambridge Structural Database 2\u00b76H2O in the molar ratio 1:1.The d(N\u2014H) = 0.96\u2005\u00c5]. All other H atoms were positioned geometrically and refined as riding, with d(C\u2014H) = 0.93\u2005\u00c5 for Csp2\u2014H and 0.97\u2005\u00c5 for methyl\u00adene C\u2014H. For all H atoms, Uiso(H) = 1.2Ueq of the parent atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901701475X/nk2241sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S205698901701475X/nk2241Isup2.hklStructure factors: contains datablock(s) I. DOI: 1579473CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The structure of a coumarin ester stabilized by C\u2014H\u22efO hydrogen bonds and C=O\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions has been studied by X-ray diffraction, Hirshfeld surface analysis and quantum chemical calculations. 16H9FO4, (I), the benzene ring is oriented at an acute angle of 59.03\u2005(15)\u00b0 relative to the coumarin plane (r.m.s deviation = 0.009\u2005\u00c5). This conformation of (I) is stabilized by an intra\u00admolecular C\u2014H\u22efO hydrogen bond, which closes a five-membering ring. In the crystal, mol\u00adecules of (I) form infinite zigzag chains along the b-axis direction, linked by C\u2014H\u22efO hydrogen bonds. Furthermore, the crystal structure is supported by \u03c0\u2013\u03c0 stacking inter\u00adactions between neighbouring pyrone and benzene or coumarin rings [centroid\u2013centroid distances in the range 3.5758\u2005(18)\u20133.6115\u2005(16)\u2005\u00c5], as well as C=O\u22ef\u03c0 inter\u00adactions [O\u22efcentroid distances in the range 3.266\u2005(3)\u20133.567\u2005(3)\u2005\u00c5]. The theoretical data for (I) obtained from quantum chemical calculations are in good agreement with the observed structure, although the calculated C\u2014O\u2014C\u2014C torsion angle between the coumarin fragment and the benzene ring (73.7\u00b0) is somewhat larger than the experimental value [63.4\u2005(4)\u00b0]. Hirshfeld surface analysis has been used to confirm and qu\u00adantify the supra\u00admolecular inter\u00adactions.In the title compound, C They also form the core of several mol\u00adecules of pharmaceutical importance. Coumarin and its derivatives have been reported to serve as anti-bacterial (Basanagouda S(5) ring motif arises from the intra\u00admolecular C16\u2014H16\u22efO3 hydrogen bond d Table\u00a01, and gend Table\u00a01. The couC(4) chains along the [010] direction = 3.336\u2005(5)\u2005\u00c5] and C1=O2\u22ef\u03c0 inter\u00adactions are present . The resulting supra\u00admolecular aggregation is completed by the presence of \u03c0\u2013\u03c0 stacking between the pyrone and C4\u2013C9 benzene rings or coumarin ring systems = 3.5758\u2005(18), Cg1\u22efCg4 = 3.6116\u2005(16), Cg2\u22efCg4 = 3.6047\u2005(16)\u2005\u00c5, where Cg2 is the centroid of the C4\u2013C9 benzene ring] are less than 3.8\u2005\u00c5, the maximum regarded as suitable for an effective \u03c0\u2013\u03c0 inter\u00adaction on ring J and distances between Cg(I) and perpendicular projection of Cg(J) on ring I (slippage) are summarized in Table\u00a02In the crystal, the C2\u2014H2\u22efO2 hydrogen bond links mol\u00adecules into infinite zigzag on Fig.\u00a02. In addims Fig.\u00a03. The cenet al., 2016et al., 1982et al., 1985et al., 2011et al., 2014meta-substituted coumarin esters to 1.20\u2005\u00c5 (blue) with the program CrystalExplorer 3.1 dnorm highlights several red spots showing distances shorter than the sum of the van der Waals radii. These dominant inter\u00adactions correspond to inter\u00admolecular C\u2014H\u22efO hydrogen bonds, C8\u22efC5 , O\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions between the surface and the neighbouring environment. The mapping also shows white or pale-red spots with distances almost equal to the sum of the van der Waals radii and blue regions with distances longer than the sum of the van der Waals radii. The surfaces are shown as transparent to allow visualization of the mol\u00adecule as predicted by the X-ray study. The most significant contrib\u00adution to the Hirshfeld surface (27.7%) is from H\u22efO/O\u22efH contacts, which appear on the left-side as blue spikes with the tip at de + di = 2.4\u2005\u00c5, top and bottom . As expected in organic compounds, the H\u22efH contacts are important with a 24.5% contribution to Hirshfeld surface; these appear in the central region of the FP with a central blue tip spike at de = di = 1.10\u2005\u00c5 whereas the F\u22efH/H\u22efF contacts with a contribution to the Hirshfeld surface of 11.4% are indicated by the distribution of points around a pair of wings at de + did). The C\u22efH/H\u22efC plot (16.2%) reveals information on the inter\u00admolecular hydrogen bonds . Other visible spots in the Hirshfeld surfaces indicate the C\u22efO/O\u22efC, O\u22efO, F\u22efF and C\u22efF/F\u22efC contacts, which contribute only 6.6, 1.3, 1.2 and 1.1%, respectively .Mol\u00adecular Hirshfeld surfaces and the associated two-dimensional fingerprint plots of (I)le Fig.\u00a04. In the le Fig.\u00a04, the adjns Fig.\u00a06a as preom Fig.\u00a06b. As ex\u2005\u00c5 Fig.\u00a06c wherea\u2005\u00c5 Fig.\u00a06d. The Cds Fig.\u00a06e. Otherly Fig.\u00a06f\u20136i.++G basis set. The crystal structure in the solid state was used as the starting structure for the calculations. The DFT calculations were performed with the GAUSSIAN09 program package 4. The resulting precipitate (crude product) was filtered off with suction, washed with petroleum ether and recrystallized from acetone. Pale-yellow crystals of (I)To a solution of 4-fluoro\u00adbenzoyl chloride in dried tetra\u00adhydro\u00adfuran (40\u2005mL) was added dried tri\u00adethyl\u00adamine and 7-hy\u00addroxy\u00adcoumarin by small portions over 30\u2005min. The mixture was then refluxed for 4\u2005h and poured into 40\u2005mL of chloro\u00adform. The solution was acidified with diluted hydro\u00adchloric acid until the pH was 2\u20133. The organic layer was extracted, washed with water to neutrality, dried over MgSOUiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901800614X/kq2021sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901800614X/kq2021Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901800614X/kq2021Isup3.cmlSupporting information file. DOI: 1834035CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the anion, the carboxyl\u00adate group is twisted away from the attached benzene ring by 84.9\u2005(2)\u00b0. The CoII atom is coordinated by two TMB anions and two water mol\u00adecules in the basal plane, while another water mol\u00adecule bridges the CoII atoms in the axial directions, forming polymeric chains running along [001]. The coordination environment for the CoII cation is a slightly distorted octa\u00adhedron. The coordinating and bridging water mol\u00adecules link to the carboxyl\u00adate groups via intra- and inter\u00admolecular O\u2014H\u22efO hydrogen bonds, enclosing S(6) ring motifs, while the coordinating, bridging and non-coordinating water mol\u00adecules link to the carboxyl\u00adate groups and the coordinating water mol\u00adecules link to the non-coordinating water mol\u00adecules via O\u2014H\u22efO hydrogen bonds, enclosing R22(8) and R33(8) ring motifs. Weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions may further stabilize the crystal structure.The asymmetric unit of the title one-dimensional polymeric compound, {[Co(C The Co1 atom lies 0.2077\u2005(1)\u2005\u00c5 above the carboxyl\u00adate (O1/O2/C1) group, which makes a dihedral angle of 84.9\u2005(2)\u00b0 with the adjacent benzene (C2\u2013C7) ring.The two carboxyl\u00adate O atoms hydrogen bonds (Table\u00a01S(6) ring motifs hydrogen bonds \u00adbenzoato-1\u03baO]mang\u00adanese(II)]-\u03bc-aqua]\u00addihydrate], {[Mn(C11H14NO2)2(H2O)3]\u00b72(H2O)}n, (II), \u00adbenzoate (DEAB) anions and two water mol\u00adecules in the basal plane, while another water mol\u00adecule bridges the Mn atoms in the axial directions, forming polymeric chains as in the title compound, (I)In the crystal structure of a similar complex, catena-\u00b72H2O}n, , {[Mn(C10H11O2)2(H2O)3]\u00b72H2O}n, (VIII), which had previously been reported by Chen et al. anions and two water mol\u00adecules in the basal plane, while another water mol\u00adecule bridges the MnII cations in the axial directions, forming polymeric chains as in the title compound, (I)We also solved the crystal structure of al. 2007. In ii distance [4.045\u2005(15)\u2005\u00c5] across the chain \u00b0] in (I)M\u2014ObrdW\u2014M bond angles increase, while the M\u2014ObrdW bond lengths decrease with increasing atomic number, Z, of the transition metal(II) atoms and the M\u22efM distances across the polymeric chains are almost the same, independent of the type of anion coordinating to the metal(II) atoms.The Co1\u22efCo1in Fig.\u00a02 and the 4\u00b77H2O with sodium 2,4,6-tri\u00admethyl\u00adbenzoate in H2O (150\u2005ml) at room temperature. The mixture was set aside to crystallize at ambient temperature for eight weeks, giving pink single crystals . FT\u2013IR: 3630, 3405, 3209, 2286, 2069, 1612, 1535, 1446, 1400, 1181, 1114, 1031, 893, 857, 827, 758, 690, 615, 570, 490, 478, 401.The title compound was prepared by the reaction of CoSOSHELXL). Bond lengths and angles for water mol\u00adecules are: O3\u2014H31 = 0.806\u2005(19), O3\u2014H32 = 0.818\u2005(18), O4\u2014H41 = 0.827\u2005(18), O5\u2014H51 = 0.812\u2005(10), O5\u2014H52 = 0.820\u2005(10)\u2005\u00c5 and H31\u2014O3\u2014H32 = 107\u2005(4) and H51\u2014O5\u2014H52 = 107\u2005(4)\u00b0 The C-bound H atoms were positioned geometrically with C\u2014H = 0.93 and 0.96\u2005\u00c5 for aromatic and methyl H atoms, respectively, and constrained to ride on their parent atoms, with Uiso(H) = k \u00d7 Ueq(C), where k = 1.5 for methyl H atoms and k = 1.2 for aromatic H atoms. The maximum and minimum electron densities were found 0.89\u2005\u00c5 and 0.82\u2005\u00c5 from Co1. The high residual electron density value of 2.178\u2005e\u2005\u00c5\u22121 may be due to the poor quality of the crystal.The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0210.1107/S2056989017005564/pj2043sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017005564/pj2043Isup2.hklStructure factors: contains datablock(s) I. DOI: 1543701CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A linear SP coordination geometry for the gold atom is found in the title structure, which also features a short intra\u00admolecular Au\u22efO inter\u00adaction, in contrast to a Au\u22ef\u03c0 inter\u00adaction found in the first polymorph. 9H9N2O3S)(C21H21P)], is a second monoclinic polymorph (space group P21/c) that complements a previously reported Cc polymorph [Broker & Tiekink \u2005\u00c5], in contrast to the aryl ring in the original polymorph. In the crystal, linear supra\u00admolecular chains along the a-axis direction mediated by C\u2013H\u22ef\u03c0 and nitro-O\u22ef\u03c0 inter\u00adactions are found. These pack with no directional inter\u00adactions between them. The analysis of the Hirshfeld surfaces for both forms of [Au(C9H9N3O3S)(C21H21P)] indicates quite distinctive inter\u00adaction profiles relating to the differences in inter\u00admolecular contacts found in their respective crystals.The title phosphanegold(I) thiol\u00adate compound, [Au(Ckink 2008. Acta Cr R3PAu[SC(OR\u2032)=NR\u2032\u2032], for R, R\u2032 = alkyl, aryl and R\u2032\u2032 = aryl, have proven to exhibit exciting biological activities. For example, compounds of the type Ph3PAu[SC(OR)=NPh], R = Me, Et and i-Pr, induce G2/M cell cycle arrest in HT-29 cancer cells and exhibit tolerable toxicity based on experiments on zebrafish thiol\u00adates of the general formula R3PAu[SC(OR\u2032)=NR\u2032\u2032] display an intra\u00admolecular Au\u22efO inter\u00adaction. In an exercise in crystal engineering, it was argued that by moderating the electronic properties of the phosphane-bound and thiol\u00adate-N-bound groups, it was possible to direct a change in conformation so that an intra\u00admolecular Au\u22ef\u03c0(ar\u00adyl) inter\u00adaction formed instead of the Au\u22efO contact =NPh], that was originally reported in a form with an intra\u00admolecular Au\u22efO inter\u00adaction =NC6H4NO2-4] is reported which was reported originally in space group Cc with a Au\u22ef\u03c0(ar\u00adyl) inter\u00adaction 3PAu[SC(OEt)=NC6H4NO2-4], (I)Cc form, (II).Referring the conformation shown in the Scheme, most structures having the formula S atom. These bond-length conclusions are vindicated by a comparison of the bond lengths found in the uncoordinated mol\u00adecule, i.e. EtOC(=S)N(H)C6H4NO2-4 Cc polymorph The central S1, O1, N1 and C1 atoms of the thiol\u00adate ligand are strictly (r.m.s. deviation of the fitted atoms = 0.0008\u2005\u00c5) planar. The plane through the nitro\u00adbenzene ligand is orthogonal to the former plane, forming a dihedral angle of 89.67\u2005(12)\u00b0. Finally, the nitro group is essentially co-planar with the ring to which it is connected, forming a dihedral angle of 4.7\u2005(4)\u00b0.Crystal Explorer a-axis direction, Fig.\u00a03a. These are sustained by a combination of nitro\u00adbenzene-C\u2014H\u22ef\u03c0(tol\u00adyl) inter\u00adactions as well as nitro-O\u22ef\u03c0(tol\u00adyl) contacts, Fig.\u00a03b. For the latter, the nitro group lies over the ring, with the two residues being almost parallel, forming a dihedral angle = 7.4\u2005(2)\u00b0. While comparatively rare, the latter inter\u00adactions have been discussed in the crystallographic literature dnorm for forms (I)a indicate the significance of short inter-atomic C\u22efO/O\u22efC contacts, Table\u00a04a, b, and the dnorm-mapped surface for (II), Fig.\u00a05c, are consistent with (I)a. The inter\u00admolecular nitro-O\u22ef\u03c0 inter\u00adaction involving both nitro\u00adbenzene-O2 and O3 atoms with the same symmetrically located tolyl ring (C17\u2013C22) are viewed as two adjoining blue and bright-orange regions in Fig.\u00a05b. The short inter-atomic S\u22efH/H\u22efS, C\u22efH/H\u22efC and O\u22efH/H\u22efO contacts influential in the structure of (II) are highlighted with black, red and yellow dashed lines, respectively, in Fig.5c.It is clearly evident from the Hirshfeld surfaces mapped over a, it is apparent that the different orientations of the thiol\u00adate ligands significantly impact upon the observed features in the plots. This is also visible from the fingerprints delineated into H\u22efH, C\u22efH/H\u22efC, O\u22efH/H\u22efO and S\u22efH/H\u22efS contacts c, wherein the half-arrows in (I)de + di \u223c\u20052.8\u2005\u00c5 and 2.9\u2005\u00c5, respectively, arise as the result of distinctive inter\u00admolecular inter\u00adactions in the two forms: the former has a C\u2014H\u22ef\u03c0 contact while the latter has short inter-atomic C\u22efH/H\u22efC contacts, Fig.\u00a05c and Table\u00a04b and Table\u00a04c and d, reflects the different types of inter-atomic contacts they form. In the respective plots for the form (I)de + di\u2005\u223c2.7\u2005\u00c5 in the O\u22efH/H\u22efO delineated and the knife-edge tips at de + di\u2005\u223c2.9\u2005\u00c5 in the S\u22efH/H\u22efS delineated fingerprint plots for (II) are the result of short inter-atomic O\u22efH/H\u22efO and S\u22efH/H\u22efS contacts, Table\u00a04The distinctive features of fingerprint plot delineated into C\u22efH/H\u22efC contacts, Fig.\u00a062)CH2(Ph2)PAuCl. In the original form, intra\u00admolecular Au\u22efAu inter\u00adactions [3.34\u2005\u00c5] were observed CH2(Ph2)PAuCl imply that Au\u22ef\u03c0(ar\u00adyl) inter\u00adactions provide comparable energies of stabilization to their crystal structures. Indeed, computational chemistry on the polymorphic system Ph3PAu[SC(OEt)=NPh] suggested the form with the intra\u00admolecular Au\u22ef\u03c0(ar\u00adyl) contact was more than 5\u2005kcal\u2005\u2005mol\u22121 stable than the form with the intra\u00admolecular Au\u22efO contact =N]2 indicated that each Au\u22ef\u03c0(ar\u00adyl) inter\u00adaction in the centrosymmetric mol\u00adecule was more stable by more than 12\u2005kcal\u2005mol\u22121 than each putative Au\u22efO contact inter\u00adactions can be seen in the polymorphic structures of ClAuP solution of (I)1H NMR (\u03b4): thiol\u00adate: 7.92 , 6.89 , 4.34 , 1.35 . Phosphane: 7.32\u20137.22 , 2.40 . 13C NMR (\u03b4): Thiol\u00adate: 165.7 , 157.5 , 142.6 , 124.8 , 122.5 , 64.5 , 14.5 . Phosphane: 142.2 , 133.9 , 129.8 , 126.4 , 21.4 .The title compound (I)Uiso(H) set to 1.2\u20131.5Ueq(C). The maximum and minimum residual electron density peaks of 1.16 and 0.78\u2005e\u2005\u00c5\u22123, respectively, were located 0.81 and 1.28\u2005\u00c5 from the Au atom. Owing to inter\u00adference from the beam-stop, the (011) reflection was omitted from the final cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989017012865/hb7703sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017012865/hb7703Isup2.hklStructure factors: contains datablock(s) I. DOI: 1573275CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the protonated piperazine ring adopts a chair conformation while the indole ring plane is nearly perpendicular to the benzo\u00adfuran ring system. 26H28N5O2+\u00b7Cl\u2212\u00b7CH3OH {systematic name: 4-(2-carbamoyl-1-benzo\u00adfuran-5-yl)-1-[4-(5-cyano-1H-indol-3-yl)but\u00adyl]piperazin-1-ium chloride methanol monosolvate}, the protonated piperazine ring adopts a chair conformation. The indole ring plane is nearly perpendicular to the benzo\u00adfuran ring system, with a dihedral angle of 85.77\u2005(2)\u00b0. In the crystal, the organic cations, Cl\u2212 anions and methanol solvent mol\u00adecules are linked by classical N\u2014H\u22efO and N\u2014H\u22efCl hydrogen bonds, and weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions into a three-dimensional supra\u00admolecular architecture.In the title compound, C Major depression disorder (MDD) currently ranks as the world\u2019s fourth greatest cause of illness and is expected to rank second by the year 2020 according to WHO studies with properties that are most similar to those of citalopram, escitalopram (levapro), fluoveline, proxetin, and sertraline. The new drug differs from its predecessors by also acting as a partial agonist at serotonergic 5-HT\u2212 anion and one methanol mol\u00adecule le Fig.\u00a01.et al., 2011n-butyl linker to the piperazine ring. The conformation of the butyl chain is of some inter\u00adest. Three C atoms of the butyl group are coplanar with atom C9 of the indole, as confirmed by the C9\u2014C10\u2014C11\u2014C12 torsion angle of 179.2\u2005(2)\u00b0, meanwhile atoms C11, C12 and C13 are coplanar with piperazine atom N3. A dihedral angle of 80.9\u2005(2)\u00b0 is formed between the mean planes of N3/C11\u2013C13 and C9\u2013C12. The dihedral angle between the C9\u2013C12 mean plane and the indole plane is 10.0\u2005(2)\u00b0. The second piperazine N atom, N4, is bonded to the benzo\u00adfuran ring. The formamide group is almost coplanar with the connected benzo\u00adfuran ring, making a dihedral angle of 2.53\u2005(2)\u00b0. The indole ring is almost perpendicular to the benzo\u00adfuran ring, as indicated by the dihedral angle of 85.77\u2005(2)\u00b0 between them.The expected proton transfer from hydro\u00adchloric acid to atom N3 of piperazine occurs; the H atom on the piperazine N3 atom was located unequivocally in the electron-density map. The six-membered piperazine ring adopts a chair conformation. The electron-withdrawing cyano group at position 5 on the indole is twisted out of the mean plane of the indole unit, as indicated by the relevant torsion angles N1\u2014C1\u2014C2\u2014C7 and N1\u2014C1\u2014C2\u2014C3 . The conformation of the cyano group is similar to that of other drugs containing nitrile groups, such as bicalutamide and Febuxostat 1\u00a0\u2212\u00a0x, \u2212y, 1\u00a0\u2212\u00a0z] hydrogen bonds 1\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y, 2\u00a0\u2212\u00a0z] and N3\u2014H3A\u22efCl1 link another two neighbouring cations and the Cl\u2212 anion into a mol\u00adecular sheet. As a result, 28-membered rings with the graph-set motif In the crystal, N3\u2014H3s Table\u00a01, connected Fig.\u00a02.Vilazodone hydro\u00adchloride was supplied by Hangzhou HEZE pharmaceutical Technology Co., Ltd. It was recrystallized from methanol solution, giving single crystals suitable for X-ray diffraction.Uiso(H) = 1.2Ueq(N). All other H atoms were placed in calculated positions with O\u2014H = 0.82, N\u2014H = 0.86 and C\u2014H = 0.93\u20130.98\u2005\u00c5, and included in the refinement in a riding model with Uiso(H) = 1.2 or 1.5Ueq(carrier atom).Experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0210.1107/S2056989016017734/xu5894sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989016017734/xu5894Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016017734/xu5894Isup3.cmlSupporting information file. DOI: 1439516CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The characteristic structural feature of a new two-dimensional Zn coordination polymer is an infinite polymeric layer parallel to the crystallographic (132) plane. 2-4,4\u2032-bis\u00ad[meth\u00adyl]biphenyl-\u03ba2N4:N4\u2032}bis\u00ad(nitrato-\u03baO)zinc(II)], [Zn(NO3)2(C18H16N6)2]n, is a two-dimensional zinc coordination polymer constructed from 4,4\u2032-bis\u00ad[meth\u00adyl]-1,1\u2032-biphenyl units. It was synthesized and characterized by elemental analysis and single-crystal X-ray diffraction. The ZnII cation is located on an inversion centre and is coordinated by two O atoms from two symmetry-related nitrate groups and four N atoms from four symmetry-related 4,4\u2032-bis\u00ad[meth\u00adyl]-1,1\u2032-biphenyl ligands, forming a distorted octa\u00adhedral {ZnN4O2} coordination geometry. The linear 4,4\u2032-bis\u00ad[meth\u00adyl]-1,1\u2032-biphenyl ligand links two ZnII cations, generating two-dimensional layers parallel to the crystallographic (132) plane. The parallel layers are connected by C\u2014H\u22efO, C\u2014H\u22efN, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions, resulting in a three-dimensional supra\u00admolecular architecture.The title compound, poly[bis\u00ad{\u03bc H-1,2,4-triazol-1-ylmeth\u00adyl)benzene benzene benzene benzene meth\u00adyl]-1,1\u2032-biphenyl meth\u00adyl]biphenyl-\u03ba2N4:N4\u2032}bis\u00ad(nitrato-\u03baO)zinc], [Zn(NO3)2(C18H16N6)2]n, was synthesized under hydro\u00adthermal conditions by the reaction of Zn(NO3)2\u00b76H2O and 4,4\u2032-bis\u00ad[meth\u00adyl]-1,1\u2032-biphenyl at 313\u2005K for 48\u2005h. We report here its crystal structure and its elemental analysis.Over the past few decades, the self-assembly of coordination polymers (CPs) or metal\u2013organic frameworks (MOFs) based on metal ions or clusters and organic ligands has attracted much attention, owing to their intriguing mol\u00adecular topologies and potential applications. Multidentate ligands derived from 1,2,4-triazole that contain an aromatic core have been used for this purpose, examples being 1,4-bis\u00admeth\u00adyl]-1,1\u2032-biphenyl ligand.The title complex crystallizes in the triclinic space group II cation exhibits a slightly distorted octa\u00adhedral {ZnN4O2} coordination geometry and is coordinated by four N atoms from four symmetry-related organic ligands and two O atoms (O3 and O3i) from two symmetry-related nitrate groups meth\u00adyl]-1,1\u2032-biphenyl ligands and two nitrate anions, and each organic ligand in turn connects two ZnII cations to generate a two-dimensional layer parallel to the crystallographic (132) plane. The organic ligand adopts a cis,cis substituent conformation. The two distinct Zn\u22efZn distances are 18.397\u2005(3) and 18.964\u2005(3)\u2005\u00c5 meth\u00adyl]-1,1\u2032-biphenyl ligand lie nearly in one plane [dihedral angle = 0.00\u2005(2)\u00b0]. The two triazole groups of the 4,4\u2032-bis\u00ad[meth\u00adyl]-1,1\u2032-biphenyl ligand are inclined to the plane of the central biphenyl groups, with dihedral angles of 80.050\u2005(2) (C1/C2/N1/N2/N3) and 85.511\u2005(2)\u00b0 (C10/C11/N4/N5/N6). Four adjacent ZnII cations are connected by four linear organic ligands and form a 72-membered macrocyclic ring in the above-mentioned two-dimensional layer \u2005\u00c5 and C12\u2014H12\u22efCg2iii = 3.5252\u2005(7)\u2005\u00c5; Cg1 and Cg2 are the centroids of the triazole (C1/C2/N1/N2/N3) and phenyl (C4\u2013C9) rings, respectively; symmetry codes: (ii) 2\u00a0\u2212\u00a0x, \u2212y, \u2212z; (iii) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, \u2212z] contacts and \u03c0\u2013\u03c0 stacking inter\u00adactions [Cg1\u22efCg1ii\u00a0= 3.6296\u2005(10)\u2005\u00c5]. These interactions, together with the covalent inter\u00adactions in the infinite two-dimensional polymeric-like layer, make up a three-dimensional supra\u00admolecular structure.Neighbouring layers are linked to each other by by weak interactions Table\u00a01, includiet al., 2016H-1,2,4-triazol-1-yl)meth\u00adyl]-1,1\u2032-biphenyl moiety gave eight hits. Seven of them are con\u00adstructed by 4,4\u2032-bis\u00ad[meth\u00adyl]-1,1\u2032-biphenyl units and different carboxyl\u00adate ligands. One example is a chain structure based on Zn and 4,4\u2032-bis\u00ad[meth\u00adyl]-1,1\u2032-biphenyl 2\u00b76H2O (0.1\u2005mmol), 4,4\u2032-bis\u00ad[meth\u00adyl]-1,1\u2032-biphenyl (0.1\u2005mmol) and water (6\u2005ml) were mixed and placed in a thick Pyrex tube, which was sealed and heated to 413\u2005K for 72\u2005h. After cooling to room temperature, colourless block-shaped crystals suitable for X-ray analysis were obtained. Elemental analysis calculated for C36H32N14O6Zn: C 52.59, H 3.92, N 23.85%; found: C 52.23, H 3.74, N 23.49%.Zn(NOUiso(H) = 1.2Ueq(C) for other H atoms. Atoms O1 and O2 of the nitrate group are disordered over two orientations, with occupancies of 0.511\u2005(11) and 0.489\u2005(11), and were refined through the use of SADI, RIGU and SIMU commands.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017012452/vn2130sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017012452/vn2130Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017012452/vn2130Isup3.molSupporting information file. DOI: 1564369CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The H atom of the oxime moiety is equally disordered over two positions, giving rise to two equivalent hydrogen bonds between adjacent mol\u00adecules. 14H13NO2, is a commercially available material and can be used as a multidentate ligand. The mol\u00adecule of the asymmetric unit has an R configuration, while the corresponding S-configured mol\u00adecule of the racemic mixture is generated by a crystallographic centre of symmetry. Both hy\u00addroxy groups are involved in hydrogen bonding, leading to the formation of chains extending parallel to [001].The title compound, C E)-2-hy\u00addroxy-1,2-diphenyl-ethan-1-one oxime, C14H13NO2, is commercially available and can be used as a multidentate ligand for which many trivial names such as cuprone or alpha-benzoin, and abbreviations including AboH2, BzoxH2, are in use. Used for a long time for the determination of manganese or copper in steel compounds, we obtained high-quality single crystals of the title compound which we have used for structure determination by X-ray diffraction.The title compound (C2/c with eight mol\u00adecules in the unit cell and one mol\u00adecule in the asymmetric unit. As the compound possesses an asymmetric carbon atom (C2), the mol\u00adecule of the asymmetric unit has an R-configuration while the corresponding S-configured mol\u00adecule of the racemic mixture is generated by a crystallographic centre of symmetry. Both mol\u00adecules also show the E configuration at the N=C double bond of the oxime moiety and sp2 (C1) hybridized C atoms. As a consequence of the different hybridization states, however, the bonds of these two carbon atoms to their phenyl groups are slightly different: 1.512\u2005(2)\u2005\u00c5 for C2 and 1.484\u2005(2)\u2005\u00c5 for C1, respectively. The hy\u00addroxy group attached to C2 shows a C\u2014O bond length of 1.425\u2005(2)\u2005\u00c5, which also lies in the normal range (1.421\u20131.433\u2005\u00c5) of a C2\u2013CH\u2013OH group \u2005\u00c5] of the N=C double bond Table\u00a01 is consiet al., 1987ar\u2014Car. The mean value of the endocyclic bond angles within the phenyl rings is 120.0\u2005(5)\u00b0 with minima at the ipso carbon atoms C11 [118.3\u2005(1)\u00b0] and C21 [119.1\u2005(1)\u00b0]. The phenyl rings form an inter\u00adplanar angle of 80.72\u2005(5)\u00b0.The two phenyl groups exhibit a mean C\u2014C bond length of 1.387\u2005(5)\u2005\u00c5 [variation: 1.374\u2005(3)\u20131.398\u2005(2)\u2005\u00c5], in excellent agreement with the literature value (Allen ii = 2.829\u2005(2)\u2005\u00c5, \u2329O2\u2014H3\u22efO2ii = 164\u00b0; symmetry code: (ii) = \u2212x\u00a0+\u00a01, \u2212y, \u2212z\u00a0+\u00a01] and a twofold rotation axis [O2\u22efO2iii = 2.806\u2005(2)\u2005\u00c5, \u2329O2\u2014H4\u22efO1iii = 175\u00b0; symmetry code (iii) = \u2212x\u00a0+\u00a01, y, \u2212z\u00a0+\u00a0i = 2.805\u2005(2)\u2005\u00c5, \u2329O1\u2014H1\u22ef N1i = 144\u00b0; symmetry code: (i) = \u2212x\u00a0+\u00a01, y, \u2212z\u00a0+\u00a0The mol\u00adecule possesses two hy\u00addroxy groups which, in principle, can act as donors and acceptors for hydrogen bonding while the N atom of the oxime moiety can only act as an acceptor atom in the formation of hydrogen bonds. In fact, the crystal packing Fig.\u00a02 with itses Fig.\u00a04. The two\u03b1-benzoinoxime was refluxed with di-n-butyl\u00adtin oxide, C8H18OSn, in ethanol for 2.5\u2005h. Single crystals of the title compound suitable for X-ray diffraction were obtained from the ethano\u00adlic solution layered with n-hexane.In a typical experiment, sp3-hybridized and 0.95\u2005\u00c5 for aromatic H atoms, and with Uiso(H) = 1.2Ueq(C). The H atoms of the two hy\u00addroxy groups were modelled with a common O\u2014H distance of 0.96\u2005\u00c5 before they were fixed and allowed to ride on the corresponding oxygen atom with Uiso(H) = 1.2Ueq(O). Disorder of the hy\u00addroxy group attached to C2 was taken into account reducing the site occupancy of both H atoms to one-half. This suggestion was confirmed by difference-Fourier maps that clearly showed both positions.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017008866/wm5397sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017008866/wm5397Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017008866/wm5397Isup3.cmlSupporting information file. DOI: 1556039CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N,N\u2010bisamide pincer ligand (ONO3\u2212) is explored. Despite their differing reactivity towards nucleophilic substrates with polarised element\u2013hydrogen bonds , both the phosphorus(III), P(ONO) (1\u2009a), and arsenic(III), As(ONO) (1\u2009b), compounds exhibit similar reactivity towards charged nucleophiles and electrophiles. Reactions of 1\u2009a and 1\u2009b with KOtBu or KNPh2 afford anionic complexes in which the nucleophilic anion associates with the pnictogen centre ([(tBuO)Pn(ONO)]\u2212 (Pn=P (2\u2009a), As (2\u2009b)) and [(Ph2N)Pn(ONO)]\u2212 (Pn=P (3\u2009a), As (3\u2009b)). Compound 2\u2009a can subsequently be reacted with a proton source or benzylbromide to afford the phosphorus(V) compounds (tBuO)HP(ONO) (4\u2009a) and (tBuO)BzP(ONO) (5\u2009a), respectively, whereas analogous arsenic(V) compounds are inaccessible. Electrophilic substrates, such as HOTf and MeOTf, preferentially associate with the nitrogen atom of the ligand backbone of both 1\u2009a and 1\u2009b, giving rise to cationic species that can be rationalised as either ammonium salts or as amine\u2010stabilised phosphenium or arsenium complexes ([Pn{ON(H)O}]+ (Pn=P (6\u2009a), As (6\u2009b)) and [Pn{ON(Me)O}]+ (Pn=P (7\u2009a), As (7\u2009b)). Reaction of 1\u2009a with an acid bearing a nucleophilic counteranion (such as HCl) gives rise to a phosphorus(V) compound HPCl(ONO) (8\u2009a), whereas the analogous reaction with 1\u2009b results in the addition of HCl across one of the As\u2212O bonds to afford ClAs{(H)ONO} (8\u2009b). Functionalisation at both the pnictogen centre and the ligand backbone is also possible by reaction of 7\u2009a/7\u2009b with KOtBu, which affords the neutral species (tBuO)Pn{ON(Me)O} (Pn=P (9\u2009a), As (9\u2009b)). The ambiphilic reactivity of these geometrically constrained complexes allows some insight into the mechanism of reactivity of 1\u2009a towards small molecules, such as ammonia and water.The ambiphilic nature of geometrically constrained Group\u200515 complexes bearing the Over the last decade significant advances have been made in the development of main\u2010group species that are capable of activating small molecules.More recently, geometrically constrained compounds based on phosphorus(III) have also received significant attention in this field. Studies by Arduengo,N,N\u2010bisamide ligand (P(ONO); 1\u2009a).1\u2009a exclusively reacts with nucleophilic species with protic E\u2212H bonds, whereas non\u2010nucleophilic substrates, even those with polar E\u2212H bonds, such as phenylsilane, fail to react. These observations seemed to indicate that the nucleophilic association of the substrate is necessary prior to any further reactivity taking place. This prompted us to explore the reactivity of 1\u2009a towards both nucleophiles and electrophiles in an effort to gain a better understanding of the reaction dynamics. These results, and analogous studies on the heavier arsenic(III) analogue (1\u2009b), are reported herein.We recently reported a phosphorus(III) compound bearing the 1\u2009a exhibits a bent geometry in the solid state with moderate pyramidalisation at both the phosphorus and nitrogen atoms (\u03a3angles(P)=296.1\u00b0; \u03a3angles(N)=331.6\u00b0), in contrast with planar compound A. Theoretical calculations at the density functional theory (DFT) level revealed that the pyramidal sC isomer (or electromorph to use the term coined by Arduengo) is the most stable, but relatively close in energy to the Cv2 symmetric species (within 4\u2005kJ\u2009mol\u22121). This energetic difference is within the error of the calculations, and it is worth noting that different computational analyses actually suggest that the planar structure is lower in energy than the sC isomer . Therefore, it is highly likely that, in solution, there is a dynamic, and concerted, pyramidal inversion at the phosphorus and nitrogen atoms resulting in a wing\u2010like \"flapping\" of the ligand backbone. The calculations also revealed that, regardless of the symmetry adopted by 1\u2009a, there is an empty, energetically accessible orbital that is largely based on the phosphorus atom, which has anti\u2010bonding character with respect to the P\u2212N and P\u2212O bonds. It is also worth noting that the P\u2212N bond in 1\u2009a is significantly polarised, with computed Hirshfeld charges of 0.394 and \u22120.171 on the phosphorus and nitrogen atoms, respectively. These observations prompted us to explore the reactivity of 1\u2009a, and its heavier arsenic analogue (1\u2009b), towards nucleophiles in order to establish whether they associate with the phosphorus/arsenic atom.As previously reported, 1\u2009b can be prepared following a similar synthetic methodology to that previously reported for its lighter congener. Reaction of the protonated ligand, N,N\u2010bisamine, H3ONO, with AsCl3 in the presence of three molar equivalents of triethylamine, affords 1\u2009b quantitatively, as evidenced by 1H and 13C{1H} NMR spectroscopy. The 1H\u2005NMR spectrum of 1\u2009b in C6D6 reveals two equal intensity aromatic resonances at 8.39 and 7.51\u2005ppm as well as two singlets at 1.71 and 1.46\u2005ppm arising from the tert\u2010butyl groups. Cooling of a concentrated pentane solution afforded bright red\u2010orange crystals of the compound in good to high yields.The arsenic\u2010containing species 1\u2009b ) of 329.1\u00b0. The sum of bond angles around the nitrogen centre, 359.9\u00b0, is significantly greater than for the lighter phosphorus\u2010containing analogue (331.6\u00b0). The As\u2212O bond lengths (1.933(4) and 1.933(4)\u2005\u00c5) are slightly elongated when compared to the expected values for single bonds (1.84\u20131.85\u2005\u00c5).1\u2009b are closely related to the planar 10\u2010As\u20103 compound 5\u2010aza\u20102,8\u2010dioxa\u2010l\u2010arsabicyclo[3.3.0]octa\u20102,4,6\u2010triene previously reported by Arduengo and co\u2010workers (As\u2212N: 1.839(3)\u2005\u00c5; As\u2212O: 1.955(3) and 1.998(3)\u2005\u00c5; \u03a3angles(As)=320.7\u00b0).1\u2009b and that determined crystallographically. Interestingly, attempts to optimise the sC isomer of 1\u2009b ultimately converge to the planar (Cv2) isomer, indicating that in contrast to its phosphorus\u2010containing analogue, the sC geometry of 1\u2009b is not a minimum on the potential energy hypersurface.The structure of b Figure\u2005 exhibits1\u2009a and 1\u2009b was explored towards a number of nucleophilic species. In a preliminary report we demonstrated that 1\u2009a reacts with nucleophiles with polarised element\u2013hydrogen bonds (NH3 and H2O) to give rise to five coordinate phosphorus(V) compounds. By contrast, no reaction is observed between 1\u2009b and ammonia, whereas hydrolysis does take place but gives rise to complex reaction mixtures and does not result in any arsenic(V)\u2010containing compounds. Our calculations reveal that this is largely a thermodynamic phenomenon. The reaction between NH3 and 1\u2009a is exothermic by 90\u2005kJ\u2009mol\u22121, whereas the same reaction between NH3 and 1\u2009b is thermodynamically uphill by 86\u2005kJ\u2009mol\u22121. This difference in reactivity can be attributed to the increased stabilisation of the \u201cinert pair\u201d on descending Group\u200515.The reactivity of 1\u2009a or 1\u2009b with neutral nucleophiles, such as pyridine or PPh3. In contrast, anionic nucleophiles do react with both compounds associating with the pnictogen centre. In a typical reaction, the geometrically constrained complexes were reacted with one equivalent of KNu (Nu=OtBu or NPh2) in the presence of a cation sequestering agent or 4,7,13,16,21,24\u2010hexaoxa\u20101,10\u2010diazabicyclo[8.8.8]hexacosane ) to aid crystallisation. These reactions yielded the anionic complexes [(tBuO)Pn(ONO)]\u2212 (Pn=P (2\u2009a) and As (2\u2009b)) and [(Ph2N)Pn(ONO)]\u2212 (Pn=P (3\u2009a) and As (3\u2009b)) as pictured in Scheme\u2005No reaction is observed for either 1\u2009a were monitored by 31P\u2005NMR spectroscopy and show quantitative conversion to the desired products, which were observed as singlet resonances at 85.5 and 66.3\u2005ppm for 2\u2009a and 3\u2009a, respectively. These resonances are shifted upfield with respect to 1\u2009a (168.6\u2005ppm), due to the enhanced electron density on the phosphorus centre. Similarly the 1H\u2005NMR spectra of both compounds display the requisite number of resonances for a symmetrical N,N\u2010bisamide ligand backbone and for the nucleophilic substituents. Reactions involving 1\u2009b similarly give rise to products in which both of the 3,5\u2010di\u2010tert\u2010butyl\u20102\u2010phenolate arms of the ligand backbone are equivalent as evidenced by 1H and 13C{1H} NMR spectroscopy.Reactions involving 2\u2009a), 0.0365\u2005\u00c5 (2\u2009b) 0.0019\u2005\u00c5 (3\u2009a) and 0.0150\u2005\u00c5 (3\u2009b)) with the nucleophile orthogonal to the Pn(ONO) core. The planarity of the ligand backbone is also evident in the sum of bond angles around the nitrogen atoms . These structures are consistent with lone\u2010pair donation from the nucleophile into the lowest unoccupied molecular orbital (LUMO) of 1\u2009a and 2\u2009a. DFT calculations reveal that the most significant atomic orbital contribution to this orbital comes from the pnictogen atom p orbital that is perpendicular to the plane of the molecule . The LUMO of 1\u2009a and 2\u2009a are also notably p\u03c0\u2013p\u03c0 antibonding with respect to the Pn\u2212N and Pn\u2212O bonds.The structures of all four novel anionic complexes were determined by single\u2010crystal X\u2010ray diffraction Figures\u2005 and 4. TN,N\u2010bisamide ligand. This significant weakening of the Pn\u2212O bonds strongly suggests that on coordination of a nucleophile, the aryloxide functionalities are susceptible to electrophilic attack. This was probed by reacting 2\u2009a and 2\u2009b with a pyridinium trifluoromethanesulfonate and benzyl bromide (BzBr).Upon coordination of an anionic nucleophile there is a significant elongation of the Pn\u2212O bonds while the Pn\u2212N bonds remain very similar to those of the parent compound or benzyl bromide afforded the trigonal bipyramidal compounds (tBuO)HP(ONO) (4\u2009a) and (tBuO)BzP(ONO) (5\u2009a), respectively to the E\u2212H activation products that were obtained by reaction of 1\u2009a with H2O and NH3. In fact, compound 4\u2009a can be readily accessed by direct reaction of 1\u2009a with HOtBu. The reactions proceeded rapidly (before the 31P\u2005NMR spectra of the reaction mixtures were recorded) at room temperature affording the compounds in quantitative yields. Consequently, we were unable to ascertain whether electrophilic association of the proton or benzyl functionalities involved the ligand backbone prior to migration of the functional group to the phosphorus centre.Reactions of [K][OTf] (Pn=P (6\u2009a), As (6\u2009b)) and [Pn{ON(Me)O}][OTf] (Pn=P (7\u2009a), As (7\u2009b)) after heating or sonicating the mixtures .31P\u2005NMR spectra of the reactions involving 1\u2009a reveal broad resonances, with evidence of weak or non\u2010existent P\u2212H coupling, at 155.5 (2JP\u2010H=12\u2005Hz) and 149.4\u2005ppm for 6\u2009a and 7\u2009a, respectively (cf. 168.6\u2005ppm for 1\u2009a), indicating that the electrophilic groups do not associate directly with the phosphorus(III) centre. The 1H\u2005NMR spectra of all four compounds are consistent with two equivalent aryloxide functionalities, which suggest functionalisation at the amide nitrogen atom. The 1H\u2005NMR resonances for the proton\u2010 and methyl\u2010group\u2010functionalised nitrogen atoms were observed at 14.27 and 3.72\u2005ppm for 6\u2009a and 7\u2009a, respectively. Similarly, reactions involving 1\u2009b also reveal clean conversion to the products and the 1H\u2005NMR resonances of the electrophiles associated with the nitrogen atom were observed at 11.64 and 3.41\u2005ppm for 6\u2009b and 7\u2009b, respectively.In a typical reaction, one molar equivalent of EOTf was added to a solution of either 6\u2009a an additional molecule of HOTf. The most striking aspect of the structures is that protonation/methylation has taken place at the nitrogen centre, as evidenced by NMR spectroscopy. All four complexes adopt a distorted tetrahedral geometry around the nitrogen atom and exhibit a significant elongation of the N\u2010Pn bonds (N\u2212P: 1.926(2) and 1.955(2)\u2005\u00c5 for 6\u2009a and 7\u2009a, respectively; N\u2010As: 2.028(2) and 2.127(1)\u2005\u00c5 for 6\u2009b and 7\u2009b, respectively) relative to 1\u2009a (1.757(1)\u2005\u00c5) and 1\u2009b (1.862(3)\u2005\u00c5). The Pn\u2212O bonds experience a moderate contraction on protonation/methylation in the case of the phosphorus containing compounds (0.03\u2005\u00c5), but a much more dramatic contraction (0.13\u2005\u00c5) for the arsenic analogues, which is linked to a change from the planar geometry of 1\u2009b to the bent (sC) geometry of both 6\u2009b and 7\u2009b. This geometric change allows for the aryloxide substituents to get closer to the arsenic centre as the pseudo\u2010axial arrangement of the oxygen atoms in the 10\u2010Pn\u20103 structure of 1\u2009b is lost on functionalisation of the ligand backbone.The structures of all four novel cationic complexes were determined by single\u2010crystal X\u2010ray diffraction Figures\u2005 and 7. T6\u2009a: 2.790(2) and 2.898(1)\u2005\u00c5, 6\u2009b: 2.745(2)\u2005\u00c5, 7\u2009a: 2.760(2) 7\u2009b: 2.226(1)\u2005\u00c5) indicating a significant degree of positive charge accumulating on the pnictogen centres on functionalisation of the ligand backbone. This is corroborated by DFT calculations which show a large Hirshfeld charges on the pnictogen atoms relative to nitrogen . Thus, all four complexes can be thought of as base\u2010stabilised phosphenium or arsenium ions. This bonding formulation has previously been proposed for related phosphorus\u2010containing compounds.Interestingly, all four species exhibit close contacts between the pnictogen(III) centre and trifluoromethanesulfonate anions (1\u2009a reacts with HOTf to afford a phosphorus(III) compound with a protonated ligand backbone (6\u2009a), the analogous reaction with an acid that has a more nucleophilic counter\u2010anion, such as HCl, affords the phosphorus(V) species HPCl(ONO) (8\u2009a). This difference in reactivity suggests that mechanistically, a sufficiently strong nucleophile is required to afford the formal phosphorus(V) oxidative addition product. Reaction of 6\u2009a with KOtBu, affords 4\u2009a, suggesting that nucleophilic association of the anionic \u2212OtBu moiety with the phosphorus centre induces proton migration from the ligand backbone. Similarly, reaction of 6\u2009a with tetradodecylammonium chloride cleanly affords 8\u2009a and the corresponding ammonium trifluoromethane sulfonate salt. Conversely, reactions of 8\u2009a with one equivalent of trimethylsilyl trifluoromethane sulfonate show evidence for the formation of 6\u2009a, although full conversion was not observed at room temperature with such stoichiometric loadings.It is interesting to note that while 1\u2009b. This is borne out in the reactivity of 1\u2009b towards HCl. Whereas 1\u2009a reacts with HCl to afford 8\u2009a, the reaction of 1\u2009b with one molar equivalent of HCl results in the addition of the acid across one of the As\u2212O bonds resulting in the arsenic(III) compound AsCl{(H)ONO} (8\u2009b). This was evident from the 1H\u2005NMR spectra of these reaction mixtures which reveal the presence of two inequivalent arms of N,N\u2010bisamide pincer ligand on protonation.In the aforementioned studies we have established the relative inaccessibility of the arsenic(V) oxidation state for 8\u2009a and 8\u2009b were both determined by single\u2010crystal X\u2010ray diffraction complexes we have studied to date, the structure of 8\u2009b clearly shows a trigonal pyramidal arsenic(III) centre in which one of the tert\u2010butyl\u20102\u2010phenolate ligand arms has been protonated and is consequently no longer coordinated to the arsenic centre. The bond metric data for 8\u2009a are similar to the other phosphorus(V) compounds reported. The P\u2212O bond lengths to the N,N\u2010bisamide ligand, 1.661(4) and 1.653(4)\u2005\u00c5, are slightly shorter than those observed for the aforementioned phosphorus(V) compounds (4\u2009a: 1.688(1) and 1.681(1); 5\u2009a: 1.721(1) and 1.732(1)\u2005\u00c5). The same is also true of the P\u2212N interatomic distance which is 1.699(5)\u2005\u00c5 in 8\u2009a, and 1.720(1) and 1.717(1)\u2005\u00c5 in 4\u2009a and 5\u2009a, respectively. The P\u2212Cl bond length is 1.918(2)\u2005\u00c5.The structures of 8\u2009b reveals a trigonal pyramidal geometry about the arsenic(III) centre (\u03a3angles(As)=284.8\u00b0; \u03a3angles(N)=355.6\u00b0) with a relatively obtuse N1\u2010As1\u2010Cl1 angle due to the steric repulsion of the chloride with the free arm of the N,N\u2010bisamide ligand. The As\u2212N and As\u2212O distances, 1.839(2) and 1.783(2)\u2005\u00c5, respectively, are notably shorter than those observed for 1\u2009b (As\u2212N: 1.862(3) and As\u2212O: 1.933(4)\u2005\u00c5), presumably due to the relaxation of steric strain and the loss of As\u2212N multiple bond character on bending the ligand backbone. It is worth noting that the As\u2212N bond in 8\u2009b is still very short and indicative of some multiple bond character.The structure of 6\u2009a with KOtBu result in the association of the tert\u2010butoxide anion with the phosphorus(III) centre and migration of the ligand proton to afford the phosphorus(V) product 4\u2009a . The facility with which the proton of the ligand backbone migrates prompted us to carry out related studies with the methylated species 7\u2009a and 7\u2009b. We hypothesised that methyl migration would not occur and that, consequently novel complexes in which both the nitrogen atom of the N,N\u2010bisamide ligand, and the heavier pnictogen centers could be functionalised.As mentioned previously, reactions of 7\u2009a or 7\u2009b were treated with one molar equivalent of potassium tert\u2010butoxide. These reactions were found to quantitively afford novel complexes bearing a symmetrical ligand environment and an additional tert\u2010butoxide functionality , two resonances arising from the ligand tert\u2010butoxide groups and a doublet due to the methyl group of the ligand backbone. Comparable spectroscopic data were recorded for the arsenic\u2010containing analogue, 9\u2009b, although this sample could not be isolated as a compositionally pure compound due to its relative instability and tendency to decompose.In a typical reaction, 9\u2009a was characterised by single\u2010crystal X\u2010ray crystallography =285.2\u00b0; \u03a3angles(N)=345.7\u00b0). That being said, the sum of bond angles around the nitrogen atom is strongly indicative of increased planarity, which we believe to arise due to the lone\u2010pair\u2013lone\u2010pair repulsion arising between the nitrogen and phosphorus centres. The amine and phosphine like character of these centres is evident in the long P\u22c5\u22c5\u22c5N interatomic distance, 2.573(2)\u2005\u00c5, which is notably longer than that observed for 6\u2009a and 7\u2009a, 1.926(2) and 1.955(2)\u2005\u00c5, respectively, and is clearly indicative of the lack of a bonding interaction between the phosphorus and nitrogen centres. The P\u2212O bonds range from 1.627(1) to 1.665(1)\u2005\u00c5 and are consistent with bond lengths reported for other phosphites.2\u2010hybridised carbon atom compared to an sp3\u2010hybridised species \u0394r=(0.03\u2005\u00c5).Compound 1\u2009a) and As(ONO) (1\u2009b) towards ionic nucleophiles and electrophiles. These studies show that anionic nucleophiles readily associate with the pnictogen(III) centres in both complexes, suggesting that such an association may play an important role in the mechanism for the bond activation of NH3 and H2O by 1\u2009a. Our studies also reveal that while phosphorus(V) compounds are readily accessible by sequential reactions involving 1\u2009a, the corresponding arsenic(V) compounds cannot be synthesised from the heavier analogue 1\u2009b.We have explored the reactivity of the geometrically constrained Group\u200515 complexes P(ONO) (\u2212). When a more nucleophilic counter\u2010anion is employed (such as Cl\u2212) these reactions result in the generation of a phosphorus(V) compound by proton migration from the ligand backbone to the phosphorus centre. As with previous studies, the analogous arsenic(V) compound was found to be inaccessible.Reactions involving charged electrophilic substrates give rise to pnictogen(III) compounds in which the electrophile associates with the nitrogen atom of the ligand backbone. Interestingly, when the electrophile in question is a proton, it will associate with the nitrogen atom of the ligand backbone only in the presence of a weakly coordinating counteranion . 1H, 13C, 31P, 19F\u2005NMR spectra were recorded at room temperature using a Bruker AVIII 500\u2005MHz or Bruker AVIII HD NanoBay 400\u2005MHz NMR Spectrometer. 1H and 13C{1H} spectra are reported relative to tetramethylsilane (TMS) and were referenced to the most downfield residual solvent resonance (1H\u2005NMR spectroscopy: 7.26\u2005ppm (CDCl3), 7.16\u2005ppm (C6D6), 5.32\u2005pm (CD2Cl2) and 3.58\u2005ppm ([D8]THF); 13C{1H} NMR spectroscopy: 77.16\u2005ppm (CDCl3), 128.06\u2005ppm (C6D6), 53.84\u2005ppm (CD2Cl2) and 67.21\u2005ppm ([D8]THF)). 31P\u2005NMR chemical shifts were externally referenced to an 85\u2009% solution of H3PO4 (aq). 19F\u2005NMR chemical shifts were externally referenced to CF3COOH. Elemental analyses were performed by Elemental analysis Ltd (Devon). EI/CI mass spectra were obtained on neat samples using a Waters GCT Time of Flight Mass Spectrometer with a temperature programmed solids probe inlet, or an Agilent 7200 Accurate\u2010Mass Q\u2010TOF GCMS with a SIM Direct Inlet probe. ESI mass spectra were obtained from DMF solutions using a Waters LCT time\u2010of\u2010flight mass spectrometer with a Z\u2010spray source .Solvents and reagents: Hexane , pentane and toluene were dried using an MBraun SPS\u2010800 solvent system, THF was dried over a potassium metal/benzophenone mixture and pyridine was distilled from CaH2. C6D6 was dried and stored over activated 3\u2005\u00c5 molecular sieves. CDCl3 , CD2Cl2 and [D8]THF were each dried over CaH2 and stored over activated 3\u2005\u00c5 molecular sieves. H3ONO and KNPh2 were synthesised according to a previously reported synthetic procedure.3 was distilled from CaH2 prior to use. AsCl3 , KOtBu , 4,7,13,16,21,24\u2010hexaoxa\u20101,10\u2010diazabicyclo\u2010hexacosane , [PyH][OTf] , HOTf , MeOTf , benzyl bromide and HCl were used as received.Synthesis of As(ONO) (1\u2009b): H3ONO was dissolved in toluene (10\u2005mL). AsCl3 was added to the stirred solution followed by NEt3 , yielding a dark orange solution and white precipitate. The mixture was stirred at room temperature for 3\u2005h. All volatiles were removed under a dynamic vacuum and the product was extracted in pentane (3\u00d710\u2005mL). Removal of volatiles in vacuo yielded 1\u2009b as a dark orange solid. Crystals suitable for single\u2010crystal X\u2010ray diffraction analysis were grown from a concentrated pentane solution at \u221230\u2009\u00b0C. Yield: 880\u2005mg (75\u2009%); elemental analysis calcd (%) for C28H40AsNO2: C 67.59, H 8.10, N 2.82; found: C 67.15, H 8.09, N 2.93; EI MS: m/z calcd for C28H40AsNO2: 497.2275; found: 497.2272; m/z calcd for C27H37AsNO2: 482.2040; found: 482.1898. 1H\u2005NMR : \u03b4=8.39 , 7.51 , 1.71 , 1.46\u2005ppm . 13C{1H} NMR : \u03b4=154.0 (Ar), 142.3 (Ar), 137.2 (Ar), 135.9 (Ar), 119.9 (Ar), 111.3 (Ar), 35.7 (C(CH3)3), 35.1 (C(CH3)3), 32.0 (C(CH3)3), 30.0\u2005ppm (C(CH3)3).tBuO)P(ONO)] ([K(18\u2010crown\u20106)][2\u2009a])Synthesis of [K(18\u2010crown\u20106)] ([K(18\u2010crown\u20106)][2\u2009b])Synthesis of [K(18\u2010crown\u20106)]\u2212: 570.29; found: 569.64; m/z calcd C28H40AsNO3 513.22; found: 513.61; 1H\u2005NMR : \u03b4=7.54 , 6.56 , 3.38 , 1.44 , 1.33 , 1.26\u2005ppm . 13C{1H} NMR : \u03b4=152.6 (Ar), 135.8 (Ar), 133.5 (Ar), 130.4 (Ar), 111.6 (Ar), 107.4 (Ar), 71.2 (OC(CH3)3), 70.8 (18\u2010crown\u20106), 35.3 (C(CH3)3), 34.9 (C(CH3)3), 32.8 (C(CH3)3), 32.8 (OC(CH3)3), 30.4\u2005ppm 3).2N)P(ONO)] ([K(18\u2010crown\u20106)][3\u2009a])Synthesis of [K(18\u2010crown\u20106)][(Ph: A preparative synthesis of [K(18\u2010crown\u20106)][3\u2009a] was not possible due to extremely high sensitivity of the product, which upon manipulation readily decomposes to [(Ph2N)HP(ONO)]. However, evidence for the formation of 3\u2009a can be observed by NMR tube scale reactions. In a typical reaction, 1\u2009a , KNPh2\u22c5THF0.15 and 18\u2010crown\u20106 were added to an NMR tube equipped with a J.\u2005Young airtight tap and the mixture dissolved in [D8]THF to give a pale yellow solution. 1H\u2005NMR : \u03b4=7.07 , 6.90\u20136.85 , 6.79\u20136.73 , 6.52\u20136.47 , 6.51 , 3.49 , 1.28 , 1.26\u2005ppm ; 13C{1H} NMR : \u03b4=151.2 , 149.4 , 135.0 (Ar), 134.8 , 129.8 , 127.6 , 125.5 , 120.0 , 112.4 (Ar), 106.7 , 70.8 , 34.9 (C(CH3)3), 24.8 (C(CH3)3), 32.5 (C(CH3)3), 30.2\u2005ppm (C(CH3)3); 31P\u2005NMR : \u03b4=66.3\u2005ppm (s); 31P{1H} NMR : \u03b4=66.3\u2005ppm (s).2N)As(ONO)] ([K(18\u2010crown\u20106)][3\u2009b])Synthesis of [K]\u2212) 513.24; found: 513.97; 1H\u2005NMR : \u03b4=7.17 , 6.76\u20136.70 , 6.50 , 6.46\u20136.41 , 3.46 , 3.43\u20133.41 , 2.46\u20132.43 , 1.36 , 1.27\u2005ppm ; 13C{1H} NMR : \u03b4=152.2 (Ar), 150.5 (NPh2), 135.8 (Ar), 134.1 (Ar), 130.6 (Ar), 127.9 (NPh2), 124.5 , 119.4 , 111.8 (Ar), 107.5 (Ar), 71.0 , 68.2 , 54.5 , 35.2 (C(CH3)3), 34.8 (C(CH3)3), 32.6 (C(CH3)3), 30.3\u2005ppm (C(CH3)3).tBuO)HP(ONO) (4\u2009a)Synthesis of and KOtBu were dissolved in THF (3\u2005mL) at room temperature. The yellow solution was stirred for 15\u2005min after which a solution of pyridinium triflate in THF (2\u2005mL) was added, giving rise to a colourless solution and white precipitate. After stirring for 30\u2005min all volatiles were removed in vacuo and the product extracted into toluene (5\u2005mL). The solution was filtered through a cannula, concentrated to approximately 0.5\u2005mL and then cooled to 5\u2009\u00b0C for a week, yielding large colourless block crystals of 4\u2009a. The product was isolated by filtration. Yield: 145\u2005mg (83\u2009%); elemental analysis calcd (%) for C32H50NO3P: C 72.83, H 9.55, N 2.65; found: C 72.92, H 9.62, N 2.75. EI MS: m/z calcd for C32H50NO3P: 528.3607; found: 528.3600 m/z calcd for C28H43NO3P: 472.2981; found: 472.2968; m/z calcd for C28H41NO2P: 454.2875; found: 454.2852; 1H\u2005NMR : \u03b4=8.16 , 7.80 , 7.22 , 1.64 , 1.42 , 1.19\u2005ppm ; 1H{31P} NMR : \u03b4=8.16\u2005ppm , all other resonances the same as above; 13C{1H} NMR : \u03b4=143.0 (Ar), 141.0 , 132.7 , 129.5 , 115.4 (Ar), 106.7 , 80.0 3), 35.0 (C(CH3)3), 34.8 (C(CH3)3), 32.0 (C(CH3)3), 30.3 3), 30.2 (C(CH3)3). 31P\u2005NMR : \u03b4=\u221238.7\u2005ppm ; 31P{1H} NMR : \u03b4=\u221238.7\u2005ppm (s).tBuO)BzP(ONO) (5\u2009a)Synthesis of and KOtBu were dissolved in THF (5\u2005mL) and stirred until all solid material had dissolved. Benzyl bromide was added, resulting in the formation of a white precipitate of KBr. The slurry was stirred for 30\u2005min and then all volatiles were removed in vacuo. The product was extracted into hexane and filtered through a cannula. The solution was concentrated under a dynamic vacuum and cooled to \u221230\u2009\u00b0C, yielding a crop of colourless crystals which were isolated by filtration. Yield: 101\u2005mg (74\u2009%); elemental analysis calcd (%) for C39H56NO3P: C 75.82, H 9.14, N 2.27; found: C 76.32, H 9.25, N 2.34; CI MS (NH3 carrier gas):\u00a0m/z calcd C35H45NO2P\u22c5NH4: 562.3688; found: 562.3479; 1H\u2005NMR : \u03b4=7.88 , 7.32\u20137.27 ), 7.22 , 6.96\u20136.90 ), 6.85\u20136.79 ), 3.78 ), 1.61 , 1.38 , 1.15\u2005ppm ; 1H{31P} NMR : 3.78\u2005ppm ) all other resonances as above; 13C{1H} NMR : \u03b4=142.9 , 141.7 (Ar), 134.8 ), 131.8 , 130.0 ), 128.8 , 128.5 ), 126.5 ), 116.1 (Ar), 106.7 , 80.6 3), 47.2 , 35.0 (C(CH3)3), 34.7 (C(CH3)3), 32.0 (C(CH3)3), 30.4 3), 30.3\u2005ppm (C(CH3)3); 31P\u2005NMR : \u03b4=\u221220.2\u2005ppm ; 31P{1H} NMR : \u03b4=\u221220.2\u2005ppm (s).Synthesis of [P{ON(H)O}][OTf] ([6\u2009a]OTf): Compound 1\u2009a (360\u2005mg 0.794\u2005mmol) was suspended in hexane (5\u2005mL) and HOTf was added, affording a colourless solution. The reaction mixture was sonicated for 15\u2005min causing precipitation of a white solid. The solid was isolated by filtration. Yield: 325\u2005mg (68\u2009%); elemental analysis calcd (%) for C29H41F3NO5PS\u22c5HCF3O3S: C 47.81, H 5.62, N 1.86; found: C 47.78, H 5.66, N 2.02. CI MS (NH3 carrier gas): m/z calcd for C28H41NO2P\u22c5NH4: 472.3219; found: 472.3002; 1H\u2005NMR : \u03b4=14.27 , 7.88 , 7.41 , 1.38 , 1.33\u2005ppm ; 13C{1H} NMR : \u03b4=149.9 (Ar), 147.0 , 138.2 (Ar), 128.9 (Ar), 126.3 (Ar), 116.5 (Ar), 35.6 (C(CH3)3), 35.4 (C(CH3)3), 31.4 (C(CH3)3), 29.4\u2005ppm (C(CH3)3); 31P\u2005NMR : \u03b4=155.5\u2005ppm ; 31P{1H} NMR : \u03b4=155.5\u2005ppm ; 19F\u2005NMR : \u03b4=\u221278.6\u2005ppm.Synthesis of [As{ON(H)O}][OTf] ([6\u2009b]OTf): Compound 1\u2009b was dissolved in hexane (5\u2005mL) and HOTf was added, resulting in a pale pink solution and the formation of a white precipitate. The mixture was stirred at room temperature for 15\u2005min after which the solid was isolated by filtration. Yield: 64\u2005mg (46\u2009%); elemental analysis calcd (%) for C28H41AsF3NO5S: C 53.78, H 6.38, N 2.16; found: C 54.33, H 6.80, N 2.21. CI MS (CH4 carrier gas): m/z calcd for C28H41AsNO2: 498.2353; found: 498.2426; m/z calcd C27H37AsNO2: 482.2027; found: 482.2133; 1H\u2005NMR : \u03b4=11.64 , 7.84 , 7.25 , 1.36 , 1.23\u2005ppm . 13C{1H} NMR : \u03b4=151.8 (Ar), 145.5 (Ar), 137.9 (Ar), 131.3 (Ar), 124.5 (Ar), 117.7 (Ar), 35.2 (C(CH3)3), 34.9 (C(CH3)3), 31.4 (C(CH3)3), 29.3\u2005ppm (C(CH3)3); 19F{1H} NMR : \u03b4=\u221278.3\u2005ppm.Synthesis of [P{ON(Me)O}][OTf] ([7\u2009a]OTf): Compound 1\u2009a was dissolved in toluene (5\u2005mL) and MeOTf was added. The solution was heated at 70\u2009\u00b0C for 24\u2005h resulting in the growth of colourless needle\u2010shaped crystals. The supernatant was removed by cannula filtration and the crystals washed with toluene (3\u00d75\u2005mL) and dried in vacuo. Yield: 85\u2005mg (60\u2009%); elemental analysis calcd (%) for C30H43F3NO5PS: C 58.33, H 7.02, N 2.27; found: C 58.50, H 7.10, N 2.35; CI MS (NH3 carrier gas): m/z calcd for C29H43NO2P\u22c5NH4: 486.3375; found: 486.3151; 1H\u2005NMR : \u03b4=7.60 , 7.39 , 3.72 , 1.39 , 1.33\u2005ppm ; 1H{31P} NMR : \u03b4=3.72\u2005ppm all other resonances as above; 13C{1H} NMR : \u03b4=149.3 (Ar), 147.0 , 139.2 (Ar), 134.0 , 126.0 (Ar), 114.8 (Ar), 47.6 , 35.6 (C(CH3)3), 35.5 (C(CH3)3), 31.5 (C(CH3)3), 29.4\u2005ppm (C(CH3)3); 31P\u2005NMR : \u03b4=149.4\u2005ppm (br\u2009s); 31P{1H} NMR : \u03b4=149.4\u2005ppm (br\u2009s); 19F\u2005NMR : \u03b4 \u221278.7\u2005ppm.Synthesis of [As{ON(Me)O}][OTf] ([7\u2009b]OTf): Compound 1\u2009b was dissolved in toluene (15\u2005mL) and MeOTf was added to the solution, which was stirred at 80\u2009\u00b0C for three days, resulting in the precipitation of a white/pink solid. The solid was isolated by filtration and dried in vacuo. Yield: 733\u2005mg (78\u2009%); elemental analysis calcd (%) for C30H43AsF3NO5S: C 54.46, H 6.55, N 2.12; found C 54.40, H 6.48, N 2.28; EI MS: m/z calcd for C29H43AsNO2\u22c5CF3O3S: 661.2030; found: 661.2022; m/z calcd for C29H43AsNO2: 512.2528; found: 512.2510; 1H\u2005NMR : \u03b4=7.31 , 7.28 , 3.41 , 1.40 , 1.31\u2005ppm ; 13C{1H} NMR : \u03b4=149.4 (Ar), 145.6 (Ar), 139.7 (Ar), 135.0 (Ar), 124.5 (Ar), 114.8 (Ar), 46.2 (NCH3), 35.5 (C(CH3)3), 34.9 (C(CH3)3), 31.6 (C(CH3)3), 29.4\u2005ppm (C(CH3)3); 19F\u2005NMR : \u03b4=\u221277.9\u2005ppm (br\u2009s).Synthesis of HPCl(ONO) (8\u2009a): Compound 1\u2009a was dissolved in Et2O (5\u2005mL) and HCl was added. The colourless solution was stirred for 30\u2005min after which all volatiles were removed in vacuo. The colourless solid was extracted into hot hexane (3\u2005mL) and the solution filtered. Colourless crystals of 8\u2009a grew upon standing at room temperature, which were isolated by filtration. A further crop could be isolated by concentrating the mother liquor. Yield: 139\u2005mg (64\u2009%); elemental analysis calcd (%) for C28H41ClNO2P: C 68.63, H 8.43, N 2.86; found: C 68.86, H 8.49, N 2.92; 1H\u2005NMR : \u03b4=8.97 , 7.66 , 7.13 , 1.45 , 1.41\u2005ppm ; 1H{31P} NMR : \u03b4=8.97\u2005ppm , all other resonances as above; 13C{1H} NMR : \u03b4=144.2 (Ar), 139.4 , 133.9 , 126.4 , 116.6 (Ar), 106.4 , 34.6 (C(CH3)3), 34.1 (C(CH3)3), 31.1 (C(CH3)3), 29.0\u2005ppm (C(CH3)3); 31P\u2005NMR : \u03b4=\u221231.3\u2005ppm ; 31P{1H} NMR : \u03b4=\u221231.3\u2005ppm (s).Synthesis of AsCl(HONO) (8\u2009b): Compound 1\u2009b was dissolved in THF (5\u2005mL). HCl was added turning the solution pale yellow. The mixture was stirred at room temperature for 15\u2005min and then concentrated in vacuo to 0.5\u2005mL promoting the growth of colourless block crystals of the product. These were isolated by filtration. Yield: 155\u2005mg (69\u2009%); elemental analysis calcd (%) for C28H41AsClNO2: C 62.98, H 7.74, N 2.62; found: C 62.64, H 7.74, N 2.96; 1H\u2005NMR : \u03b4=7.53 , 7.41 , 7.19 , 6.69 , 5.48 , 1.63 , 1.57 , 1.27 , 1.16\u2005ppm ; 13C{1H} NMR : \u03b4=149.7 (Ar), 147.9 (Ar), 146.1 (Ar), 143.5 (Ar), 137.4 (Ar), 137.2 (Ar), 136.7 (Ar), 124.6 (Ar), 124.0 (Ar), 123.6 (Ar), 116.5 (Ar), 108.5 (Ar), 35.6 (C(CH3)3), 35.1 (C(CH3)3), 34.8 (C(CH3)3), 34.6 (C(CH3)3), 31.7 (C(CH3)3), 31.6 (C(CH3)3), 30.0 (C(CH3)3), 29.8\u2005ppm (C(CH3)3).tBuO)P{ON(Me)O} (9\u2009a)Synthesis of and KOtBu were dissolved in THF (3\u2005mL) at \u221278\u2009\u00b0C and the reaction mixture was allowed to warm to room temperature. After one hour all volatiles were removed in vacuo. The product was extracted with hexane (3\u2005mL) and the solution was concentrated under a dynamic vacuum and cooled to 5\u2009\u00b0C, resulting in the formation of a colourless microcrystalline solid. Crystals suitable for X\u2010ray diffraction studies were grown from a concentrated hexane solution at room temperature. Yield: 72\u2005mg (50\u2009%); elemental analysis calcd (%) for C33H52NO3P: C 73.16, H 9.68, N 2.59; found: C 73.04, H 9.48, N 2.68. CI MS (NH3 carrier gas): m/z calcd for C29H44NO3P\u22c5H: 486.3137; found: 486.3140; m/z calcd for C29H43NO2P: 468.3026; found: 468.3051; 1H\u2005NMR : \u03b4=7.40 , 7.31 , 2.89 , 1.57 1.57 , 1.25\u2005ppm ; 13C{1H} NMR : \u03b4=149.7 , 144.6 (Ar), 139.3 (Ar), 138.9 , 121.5 (Ar), 118.6 (Ar), 75.7 3), 42.5 , 35.5 (C(CH3)3), 34.7 (C(CH3)3), 31.8 (OC(CH3)3), 31.7 (C(CH3)3), 30.1\u2005ppm (C(CH3)3); 31P\u2005NMR \u03b4=142.8\u2005ppm (s); 31P{1H} NMR \u03b4=142.8\u2005ppm (s).tBuO)As{ON(Me)O} (9\u2009b)Synthesis of was dissolved in THF (5\u2005mL) and cooled to \u221278\u2009\u00b0C. A solution of KOtBu in THF (5\u2005mL) was then added dropwise. The reaction mixture was stirred at \u221278\u2009\u00b0C for 30\u2005mins, then 30\u2005mins at room temperature. All volatiles were removed in vacuo and the product was extracted with hexane (5\u2005mL) and filtered via cannula. The solution was concentrated under a dynamic vacuum yielding a crude sample of 9\u2009b as a white solid. Isolation of 9\u2009b from other impurities has not been possible, although characterisation data confirms the presence of the expected product. EI MS: m/z calcd for C33H53AsNO3: 586.3241; found: 586.3140; m/z calcd for C29H44AsNO3 [(HO)As{ON(Me)O}]: 529.2537; found: 529.2534; m/z calcd for C29H43AsNO2 [As{ON(Me)O}]: 512.2510\u2009; found: 512.2500; 1H\u2005NMR : \u03b4=7.40 , 7.32 , 2.62 , 1.59 1.54 , 1.28\u2005ppm ; 13C{1H} NMR : \u03b4=151.4 (Ar), 142.9 (Ar), 139.0 (Ar), 137.9 (Ar), 121.9 (Ar), 118.0 (Ar), 73.6 (OC(CH3)3), 43.0 (NCH3), 35.7 (C(CH3)3), 34.6 (C(CH3)3), 33.1 (OC(CH3)3), 31.7 (C(CH3)3), 30.1\u2005ppm (C(CH3)3).X\u2010ray diffraction: Single\u2010crystal X\u2010ray diffraction data were collected using an Oxford Diffraction Supernova dual\u2010source diffractometer equipped with a 135\u2005mm Atlas CCD area detector. Crystals were selected under Paratone\u2010N oil, mounted on micromount loops and quench\u2010cooled using an Oxford Cryosystems open flow N2 cooling device.\u03bb=1.5418\u2005\u00c5; Oxford Diffraction Supernova). Data were processed using the CrysAlisPro package, including unit cell parameter refinement and inter\u2010frame scaling (which was carried out using SCALE3 ABSPACK within CrysAlisPro).F2 using the SHELXL 2013\u20134 package.CCDC\u20051489274 (1\u2009b), 1489275 ][2\u2009a]\u22c51.5\u2009toluene), 1489276 ([K(18\u2010crown\u20106)][2\u2009b]\u22c5THF), 1489277 ([K(18\u2010crown\u20106)][3\u2009a]\u22c50.5\u2009toluene\u22c50.5\u2009pentane), 1489278 ][3\u2009b]), 1489279 (4\u2009a), 1489280 (5\u2009a), 1489281 ([6\u2009a][OTf]\u22c5HOTf), 1489282 ([6\u2009b][OTf]\u22c50.5\u2009hexane), 1489283 ([7\u2009a][OTf]), 1489284 ([7\u2009b][OTf]), 1489285 (8\u2009a), 1489286 (8\u2009b), and 1489287 (9\u2009a) contain the supplementary crystallographic data. These data can be obtained free of charge by The Cambridge Crystallographic Data Centre.DFT calculations: All geometry optimisations were performed using the Amsterdam Density Functional package (ADF2014.01).As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "Except for van der Waals forces, there are no significant inter\u00admolecular inter\u00adactions in the crystal.The mol\u00adecular structure of the title titanium(IV) half-sandwich complex, [Ti(\u03b7 Structural details based on the results of X-ray diffraction and of density functional theory calculations at the M06-2X level support the formulation of these complexes as non-classical mono\u00adaza\u00adbutadiene complexes. However, the follow-up chemistry with various multiple bond substrates of the complexes with formulae [(\u03b75-Cp#)Ti(\u03b74-C21H19N)(Cl)] shows a hidden \u03b72-imine reactivity to five-membered titanacycles Ti(\u03b74-C21H19N)(Cl)] with the dialkyl-substituted lithium amide LiN(Me)Cy, the formation of a titanadi\u00adhydro\u00adpyrrole is observed as a result of the 1,3-H-shift in the five-membered ring system in addition to the salt metathesis reaction Ti(\u03b74-C21H19N)(C12H10N), 1, synthesized by the reaction of [(\u03b75-Cp*)Ti(\u03b74-C21H19N)(Cl)] with the diaryl-substituted lithium amide LiNPh2. Compound 1 maintains the \u03b74-coordination mode of the ketimine ligand.Here we report the synthesis and crystal structure of the title compound (\u03b71 for which the \u03b74-coordination mode of the ketimine ligand is clearly confirmed. The N1\u2014C17 bond length [1.383\u2005(3)\u2005\u00c5] is significantly elongated compared to the free ketimine in contrast to the well-balanced C\u2014-C distances in the C18\u2013C23 aromatic ring system (\u2243 1.39\u2005\u00c5). The central titanium(IV) atom is fourfold coordinated in a considerably distorted tetra\u00adhedral coordination environment, with N1\u2014Ti1\u2014N2 and N1\u2014Ti1\u2014C30 bond angles of 110.42\u2005(9) and 84.23\u2005(9)\u00b0, respectively. The Ti1\u2014N1 bond length [1.963\u2005(2)\u2005\u00c5] is shorter than the Ti1\u2014N2 bond length [2.009\u2005(2)\u2005\u00c5] and indicates weak \u03c0p\u2013\u03c0d electron donor inter\u00adactions. The Ti1\u2014C30 bond length [2.259\u2005(3)\u2005\u00c5] as well as the fold angle of the central five-membered ring system (60.6\u00b0) are similar to those in other reported mono\u00adaza\u00adbutadiene complexes /2 \u2013 (Ti1\u2014N1 + Ti1\u2014C30)/2] = 0.386\u2005\u00c5 Ti(\u03b74-C21H19N)(Cl)] and lithium diphenyl amide were prepared according to published procedures Ti(\u03b74-C21H19N)(Cl)] and lithium diphenyl amide were dissolved in 12\u2005ml of tetra\u00adhydro\u00adfuran. After stirring the reaction mixture for 16\u2005h at room temperature, the solvent was evaporated in a vacuum. The residue was dissolved in 12\u2005ml of toluene, filtered, and the precipitate of LiCl was washed with toluene (2 \u00d710\u2005ml). The combined filtrates were evaporated in a vacuum and the residue was recrystallized from n-hexane to yield complex 1 as dark-red prisms in 15% crystalline yield.[ I. DOI: 10.1107/S2056989017017455/wm5424Isup2.hklStructure factors: contains datablock(s) I. DOI: 1589353CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A and B) with similar conformations. In the crystal, they are arranged alternately, forming \u2013A\u2013B\u2013A\u2013B\u2013 chains linked by N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds, which extend along the a-axis direction.The asymmetric unit of the title carbamate, contains two independent mol\u00adecules . The di\u00admethyl\u00adphenyl ring, the phenyl ring and the central carbamate N\u2014C(=O)\u2014O group are not coplanar. In mol\u00adecule A, the di\u00admethyl\u00adphenyl and phenyl rings are inclined to the carbamate group mean plane by 27.71\u2005(13) and 71.70\u2005(4)\u00b0, respectively, and to one another by 84.53\u2005(13)\u00b0. The corresponding dihedral angles in mol\u00adecule B are 34.33\u2005(11), 66.32\u2005(13) and 85.48\u2005(12)\u00b0, respectively. In the crystal, the A and B mol\u00adecules are arranged alternately linked through N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds, forming \u2013A\u2013B\u2013A\u2013B\u2013 chains, which extend along [100]. Within the chains and linking neighbouring chains there are C\u2014H\u22ef\u03c0 inter\u00adactions present, forming columns along the a-axis direction. The columns are linked by offset \u03c0\u2013\u03c0 stacking inter\u00adactions, forming a three-dimensional network [shortest centroid\u2013centroid distance = 3.606\u2005(1)\u2005\u00c5].The asymmetric unit of the title compound, C In mol\u00adecule A, the di\u00admethyl\u00adphenyl ring (C1\u2013C6) makes a dihedral angle of 84.53\u2005(13)\u00b0 with the phenyl ring (C10\u2013C15), and in mol\u00adecule B the di\u00admethyl\u00adphenyl ring (C16\u2013C21) makes a dihedral angle of 85.48\u2005(12)\u00b0 with the phenyl ring (C25\u2013C30). In mol\u00adecule A, the aryl rings (C1\u2013C6 and C10\u2013C15) are inclined to the the mean plane of the carbamate N1\u2014C9(=O2)\u2014O1 unit by 27.71\u2005(13) and 71.70\u2005(14)\u00b0, respectively. In mol\u00adecule B, rings C16\u2013C21 and C25\u2013C39 are inclined to the the mean plane of the carbamate N2\u2014C24(=O24)\u2014O13 unit by 34.33\u2005(11) and 66.32\u2005(13)\u00b0, respectively. The C9\u2014N1 and C24\u2014N2 distances are 1.336\u2005(3) and 1.335\u2005(3)\u2005\u00c5, respectively, indicating partial double-bond character in the carbamate unit.The asymmetric unit of the title compound, Fig.\u00a01A\u2013B\u2013A\u2013B\u2013 chains, propagating along the a-axis direction . These inter\u00adactions form columns along the a-axis direction, which are linked by offset \u03c0\u2013\u03c0 stacking inter\u00adactions , forming a three-dimensional network, as illustrated in Fig.\u00a04Cg1\u22efCg1iii = 3.738\u2005(2)\u2005\u00c5, inter\u00adplanar distance = 3.521\u2005(1)\u2005\u00c5, slippage = 1.257\u2005\u00c5; Cg3\u22efCg3iv = 3.606\u2005(1)\u2005\u00c5, inter\u00adplanar distance = 3.462\u2005(1)\u2005\u00c5, slippage = 1.007\u2005\u00c5; Cg1 and Cg3 are the centroids of the C1\u2013C6 and C16\u2013C21 rings, respectively; symmetry codes: (iii) \u2212x\u00a0+\u00a03, \u2212y, \u2212z\u00a0+\u00a01; (iv) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01].In the crystal, N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds link the mol\u00adecules to form \u2013n Table\u00a01. Within ns Fig.\u00a03b, formiet al., 2016Pna21 polymorphs of phenyl phenyl\u00adcarbamate itself, viz. YEHPOQ and 85.5\u2005(1)\u00b0, respectively, in mol\u00adecules A and B of the title compound.A search of the Cambridge Structural Database of 3,5 dimethyl aniline dissolved in 100\u2005ml of dry THF was added a calculated 5% excess of phenyl\u00adchloro\u00adforamate in 50\u2005ml of dry THF. The addition rate was such that it took 1.5\u2005h for complete transfer. After the addition was complete, stirring was continued overnight. Excess THF was removed under vacuum at room temperature. The crude product was extracted with ethyl acetate (3 \u00d7 100\u2005ml), and then the organic layer was dried over anhydrous sodium sulfate. Removing the solvent under vacuum at room temperature, yielded a light-yellow product which was dried under vacuum to constant weight. Yellow block-like crystals were obtained by slow evaporation of an ethyl acetate solution at room temperature (yield 99%).Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq for the H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017006922/su5370sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989017006922/su5370Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017006922/su5370Isup3.cmlSupporting information file. DOI: 1548793CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Supra\u00admolecular layers in the bc plane sustained by C\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions feature in the crystal.The title mol\u00adecule has a approximately coplanar relationship between the methyl\u00adidenehydrazinecarbodi\u00adthio\u00adate (C 16H17N3S2 (systematic name: (Z)-{[(benzyl\u00adsulfan\u00adyl)methane\u00adthio\u00adyl]amino}[1-(6-methyl\u00adpyridin-2-yl)ethyl\u00adidene]amine), the central methyl\u00adidenehydrazinecarbodi\u00adthio\u00adate (C2N2S2) core is almost planar (r.m.s. deviation = 0.0111\u2005\u00c5) and forms dihedral angles of 71.67\u2005(3)\u00b0 with the approximately orthogonally inclined thio\u00adester phenyl ring, and 7.16\u2005(7)\u00b0 with the approximately coplanar substituted pyridyl ring. The latter arrangement and the Z configuration about the imine-C=N bond allows for the formation of an intra\u00admolecular hydrazine-N\u2014H\u22efN(pyrid\u00adyl) hydrogen bond that closes an S(6) loop. In the crystal, phenyl-C\u2014H\u22efS(thione), methyl\u00adene-C\u2014H\u22ef\u03c0(pyrid\u00adyl), methyl\u00adene- and phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl) contacts connect mol\u00adecules into supra\u00admolecular layers propagating in the bc plane; the layers stack along the a axis with no directional inter\u00adactions between them. The analysis of the Hirshfeld surface indicates the relative importance of an intra\u00adlayer phenyl-H\u22efH(pyrid\u00adyl) contact upon the mol\u00adecular packing.In the title di\u00adthio\u00adcarbazate ester, C Different ligands can be obtained by introducing different organic substituents, which causes variation in their biological properties, although they may differ only slightly in their mol\u00adecular structures and 7.16\u2005(7)\u00b0, respectively, indicating nearly orthogonal and co-planar dispositions, respectively; the dihedral angle between the outer rings is 65.79\u2005(4)\u00b0. The configuration about the imine-C9=N2 bond [1.2924\u2005(18)\u2005\u00c5] is Z, resulting in the hydrazine-N1\u2014H hydrogen atom being directed towards the pyridyl-N3 atom, enabling the formation of an inter\u00admolecular amine-N1\u2014H\u22efN3(pyrid\u00adyl) hydrogen bond that closes an S(6) loop, Table\u00a01syn with the thione-S1 atom and at the same time is orientated to the opposite side of the mol\u00adecule to the imine-bound methyl group.The mol\u00adecular structure of (I)b-axis direction, and a number of C\u2014H\u22ef\u03c0 contacts, i.e. methyl\u00adene-C2\u2014H\u22ef\u03c0(pyrid\u00adyl), phenyl-C7\u2014H\u22ef\u03c0(phen\u00adyl) and methyl-C10\u2014H\u22ef\u03c0(phen\u00adyl), as detailed in Table\u00a01bc plane, Fig.\u00a02a. Layers stack along the a axis with no directional inter\u00adactions between them, Fig.\u00a02b.The participation of the hydrazine-N1\u2014H hydrogen and pyridyl-N3 atoms in the intra\u00admolecular N\u2014H\u22efN hydrogen bond precludes their participation in inter\u00admolecular inter\u00adactions. The mol\u00adecular packing features weak phenyl-C8\u2014H\u22efS2(thione) inter\u00adactions, leading to chains along the et al., 2017dnorm in Fig.\u00a03a (labelled as \u20181\u2019). The presence of the weak inter\u00admolecular C\u2014H\u22efS contact involving the phenyl-C8 and thione-S2 atoms is evident from the diminutive red spots near these atoms in Fig.\u00a03b (labelled as \u20183\u2019) characterize the short surface C\u22efH/H\u22efC contacts and indicate the relative importance of this particular C\u2014H\u22ef\u03c0 contact compared with the other two C\u2014H\u22ef\u03c0 contacts summarized in Table\u00a01i.e. \u03c0\u22efH\u2014C, containing \u03c0-bond acceptors, on the Hirshfeld surface mapped with the shape-index property are illustrated in Fig.\u00a05The Hirshfeld surfaces calculated for (I)a, and those delineated indicate the significant influence of the short inter\u00adatomic phenyl-H\u22efH(pyrid\u00adyl) contacts in the crystal mentioned above. The inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions discussed earlier are characterized by short inter\u00adatomic C\u22efH/H\u22efC contacts and pyridyl rings on the Hirshfeld surface mapped over the electrostatic potential in Fig.\u00a04de + di \u223c 2.8\u2005\u00c5 in Fig.\u00a06d. The inter\u00adatomic N\u22efH/H\u22efN contacts do not represent directional inter\u00adactions as the inter\u00adatomic separations are greater than sum of their van der Waals radii as evident from Fig.\u00a06e. Similarly, the other surface contacts summarized in Table\u00a03The overall two-dimensional fingerprint plot for (I)ts Fig.\u00a06b indicas Table\u00a02 and theiChemical context, there is sustained inter\u00adest in this class of compound and this is reflected by the observation there are four closely related structures available for comparison, varying in the S-bound group and substitution in the 2-pyridyl ring. In common with (I)Z-configuration is noted about the imine bond allowing for the formation of an intra\u00admolecular hydrazine-N\u2014H\u22efN(pyrid\u00adyl) hydrogen bond hydrogen bonds and eight-membered {\u22efHNCS}2 synthons.As mentioned in the S-Benzyl\u00addithio\u00adcarbazate (SBDTC) was prepared according to the method published by Ali & Tarafder (1977\u22121): 2921 \u03bd(N\u2014H), 1560 \u03bd(C=N), 1055 \u03bd(N\u2014N), 881 \u03bd(CSS).All chemicals were of analytical grade and were used without any further purification. fder 1977. PotassiUiso(H) set to 1.2\u20131.5Ueq(C). The nitro\u00adgen-bound H atom was located in a difference Fourier map, but was refined with a distance restraint of N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989018001330/hb7730sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018001330/hb7730Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018001330/hb7730Isup3.cmlSupporting information file. DOI: 1818384CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The 5FC ring forms a dihedral angle of 19.97\u2005(11)\u00b0 with the ring of the picrate (PA\u2212) anion. In the crystal, the 5FC+ cation inter\u00adacts with the PA\u2212 anion through three-centre N\u2014H\u22efO hydrogen bonds, forming two conjoined rings having R21(6) and R12(6) motifs, and is extended by N\u2014H\u22efO hydrogen bonds and C\u2014H\u22efO inter\u00adactions into a two-dimensional sheet structure lying parallel to (001). Also present in the crystal structure are weak C\u2014F\u22ef\u03c0 inter\u00adactions.In the crystal structure of the title compound, 5-fluoro\u00adcytosinium picrate, C New solid forms of pharmaceuticals are designed using the crystal engineering approach. These engineered solids have technological and legal importance. Among the inter\u00admolecular inter\u00adactions, hydrogen bonding is the master key for mol\u00adecular recognition in biological systems because of its strength and directionality and one picrate anion (PA\u2212) anion, the nitro groups lie variously out of the parent benzene ring, with torsion angles C9\u2014C8\u2014N5\u2014O4, C9\u2014C10\u2014N6\u2014O7 and C11\u2014C12\u2014N7\u2014O9 of 166.2\u2005(2), \u2212171.7\u2005(2) and 147.2\u2005(2)\u00b0, respectively.The asymmetric unit contains one 5-fluoro\u00adcytosinium cation (5FC\u2212) Fig.\u00a01. The 5-f+ cation inter\u00adact with atoms O3 and O9 of the picrate anion through three-centre N\u2014H\u22efO hydrogen bonds, forming two fused-ring motifs with graph-sets + cation acts as a three-centre donor and the O3 atom of the PA\u2212 anion acts as a three-centre acceptor. This type of inter\u00adaction has also been reported in the crystal structures of 2-amino-4,6-di\u00admethyl\u00adpyrimidinium picrate Fig.\u00a01. One of g Table\u00a01. A similii atom and the amino group of the 5FC+ cation are connected through an N\u2014H\u22efO hydrogen bond, forming a two-dimensional supra\u00admolecular network lying parallel to (001) \u2005\u00c5; C\u2014F\u22efCg = 88.34\u2005(12)\u00b0, where Cg is the centroid of the N1\u2013C6 ring; symmetry code: (iv) \u2212x, \u2212y, \u2212z\u00a0+\u00a01]. A similar angle [90.5\u2005(2)\u00b0] has been reported for a C\u2014F\u22efCg inter\u00adaction in an acridinium tri\u00adfluoro\u00admethane sulfonate compound Fig.\u00a02. Also pr1) Fig.\u00a02 between et al., 1982et al., 2013et al., 2012et al., 2014et al., 2001et al., 2014et al., 2016et al., 2006et al., 2009et al., 1995et al., 2004et al., 2004The crystal structures of 5-fluoro\u00adcytosine monohydrates (Louis A hot aqueous solution of 5-fluoro\u00adcytosine (32\u2005mg) and picric acid (57\u2005mg) were mixed in a 1:1 molar ratio. The resulting solution was warmed to 353\u2005K wrong symmetry description - inversion centre in central benzene ring over a water bath for half an hour and kept for slow evaporation. After a week, colourless prismatic crystals were obtained.Uiso(H) = 1.2Ueq(parent atom).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901700216X/zs2375sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901700216X/zs2375Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901700216X/zs2375Isup3.cmlSupporting information file. DOI: 1531927CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the crystal of the title compound contains two rivaroxaban mol\u00adecules with different conformations. 19H18ClN3O5, contains two rivaroxaban mol\u00adecules with different conformations; the C\u2014C\u2014N\u2014C torsion angles between the oxazolidine and thio\u00adphene rings are \u2212171.1\u2005(7) and \u2212106.8\u2005(9)\u00b0 in the two independent mol\u00adecules. In the crystal, classical N\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular architecture.The asymmetric unit of the crystal of the title compound (common name rivaroxaban), C At present, the incidence of thromboic disease is extremely high; this is mainly caused by vascular endothelial injury, increased blood coagulation, increased platelet number and decreased anti\u00adcoagulant activity and \u2212106.8\u2005(9)\u00b0, respectively \u2005\u00c5 for the O2A atom], whereas the oxazolidine ring of mol\u00adecule B displays an envelope conformation with atom C8B as the flap. The morpholine rings of the two mol\u00adecules display similar twisted boat conformations. Atoms O4 and C17 deviate from the C16/N3/C19/C18 mean plane by 0.230\u2005(2) and 0.517\u2005(2)\u2005\u00c5, respectively, in mol\u00adecule A and by 0.290\u2005(2) and 0.489\u2005(2)\u2005\u00c5 in mol\u00adecule B.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01y Table\u00a01. The oxaA and B into dimers, and weak C\u2014H\u22efO hydrogen bonds link the dimers to form a three-dimensional supra\u00admolecular architecture . The O\u22efH/H\u22efO inter\u00adactions (22.4%), which are the most significant inter\u00admolecular inter\u00adactions and link the mol\u00adecular dimers into infinite chains along the b axis, appear as two obvious spikes . At the top left (di < de) and bottom right (di > de) of the fingerprint plot, there are characteristic \u2018wings\u2019 that are identified resulting from the C\u22efH/H\u22efC inter\u00adactions (18.7%) shown in Fig.\u00a03d.The light-red spots on the Hirshfeld surface are the results of N\u2014H\u22efO, C\u2014H\u22efO and C\u2014Cl\u22efO inter\u00adactions Fig.\u00a03. The H\u22efHes Fig.\u00a03c. At thThe crude product was supplied by the Zhejiang Huadong Pharmaceutical Co., Ltd. It was recrystallized from methanol solution, giving colourless crystals suitable for X-ray diffraction.A and H1B were found in difference-Fourier maps, but placed in calculated positions with N\u2014H = 0.86\u2005\u00c5 and refined as riding with Uiso(H) = 1.2Ueq(N). All other H atoms were placed in calculated positions with C\u2014H = 0.93\u20130.98\u2005\u00c5 and included in the refinement in a riding model, with Uiso(H) = 1.2 or 1.5Ueq(carrier atom).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017017819/xu5913sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989017017819/xu5913Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017017819/xu5913Isup3.cmlSupporting information file. DOI: 1810879CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal of (I), supra\u00admolecular layers parallel to (10-1) are sustained by methyl\u00adene-C\u2014H\u22efO(meth\u00adoxy) inter\u00adactions, while in (II), centrosymmetric dimers are formed via pairwise weak phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl) contacts.The coordination geometry in ( 4H9)2(C7H14NO2S2)2], (I), and [Sn(C6H5)3(C5H10NOS2)], (II), are described. Both structures feature asymmetrically bound di\u00adthio\u00adcarbamate ligands leading to a skew-trapezoidal bipyramidal geometry for the metal atom in (I) and a distorted tetra\u00adhedral geometry in (II). The complete mol\u00adecule of (I) is generated by a crystallographic twofold axis (Sn site symmetry 2). In the crystal of (I), mol\u00adecules self-assemble into a supra\u00admolecular array parallel to (10-1) via methyl\u00adene-C\u2014H\u22efO(meth\u00adoxy) inter\u00adactions. In the crystal of (II), supra\u00admolecular dimers are formed via pairs of weak phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl) contacts. In each of (I) and (II), the specified assemblies connect into a three-dimensional architecture without directional inter\u00adactions between them. Hirshfeld surface analyses confirm the importance of H\u22efH contacts in the mol\u00adecular packing of each of (I) and (II), and in the case of (I), highlight the importance of short meth\u00adoxy-H\u22efH(but\u00adyl) contacts between layers.The crystal and mol\u00adecular structures of the two title organotin di\u00adthio\u00adcarbamate compounds, [Sn(C The coordination environment is completed by two \u03b1-carbon atoms of the n-butyl substituents. The resultant C2S4 donor set defines a skew-trapezoidal bipyramidal geometry with the tin-bound organic substituents lying over the weaker Sn\u2014S2 bonds, which subtend an angle at the tin atom approximately 50\u00b0 wider than that subtended by the S1 atoms, Table\u00a012CN residue and have very similar conformations, as seen in the values of the C1\u2014N1\u2014C2\u2014C3, N1\u2014C2\u2014C3\u2014O1 and C2\u2014C3\u2014O1\u2014C4 torsion angles of \u221294.1\u2005(4), \u221267.4\u2005(4) and \u2212177.1\u2005(3)\u00b0, indicating that \u2212 anti-clinal, \u2212 syn-clinal and \u2212 anti-periplanar descriptors, respectively, are in effect. For the O2-meth\u00adoxy\u00adethyl group, the equivalent torsion angles are \u221282.0\u2005(4), \u221270.3\u2005(4) and \u2212169.1\u2005(3)\u00b0. The independent n-butyl substituent has an all-trans (+ anti-periplanar) conformation, as seen in the values of the Sn\u2014C8\u2014C9\u2014C10 and C8\u2014C9\u2014C10\u2014 C11 torsion angles of 172.9\u2005(2) and 176.3\u2005(3)\u00b0, respectively.The tin atom in (I)b, lies on a general position and has a quite distinct coordination geometry owing to the presence of three tin-bound phenyl groups. As for (I)cf. (I)i.e. nearly 0.07\u2005\u00c5, Table\u00a02cf. (I)3S2 donor set approximates a trigonal\u2013bipyramidal geometry with the value of \u03c4, an indicator of a five-coordinate coordination geometry, being 0.61, cf. 1.0 for an ideal trigonal bipyramid and 0.0 for an ideal square pyramid \u00b0, for S1\u2014Sn\u2014C11, to 119.09\u2005(7)\u00b0, for S1\u2014Sn\u2014C31. The C21\u2014Sn\u2014C31 angle, at 115.55\u2005(10)\u00b0, is wider by 10\u00b0 than the other C\u2014Sn\u2014C angles, a result correlated with the close approach of the S2 atom. The 2-meth\u00adoxy\u00adethyl group has a very similar conformation to the O1-meth\u00adoxy\u00adethyl group in (I)The mol\u00adecule in (II)b so that a supra\u00admolecular array is formed parallel to PLATON et al., 2017The Hirshfeld surface calculations for the organotin derivatives (I)A and H6B atoms lying on both the sides of twofold symmetry axis on the Hirshfeld surfaces mapped over dnorm for (I)a and b represent the dominant inter\u00admolecular C\u2014H\u22efO contacts, Table\u00a03B and butyl-H8A atoms in Fig.\u00a04c indicate the significant influence of intra-layer H\u22efH contacts, Table\u00a05a and b, the donors and acceptors are represented with blue and red regions around the respective atoms corresponding to positive and negative potentials, respectively.The bright-red spots near each of the meth\u00adoxy-O1 and -O2, and methyl\u00adene-H4dnorm for (II)c are due to polarization of charges near the respective functional groups and do not represent any significant inter\u00adaction in the crystal. The weak inter\u00admolecular C\u2014H\u22ef\u03c0 contact and intra-layer inter\u00adatomic H\u22efH contacts et al., 2007de + di \u223c 2.0\u2005\u00c5 and the distribution of points with greater density in range \u223c1.0 to 1.2\u2005\u00c5 for (I)a, and a small peak at de + di \u223c 2.2\u2005\u00c5 with relatively few points at de + di < 2.4\u2005\u00c5 for (II)b. The fingerprint plot delineated into O\u22efH/H\u22efO contacts for (I)a, characterizes inter\u00admolecular C\u2014H\u22efO inter\u00adactions as the pair of forceps-like tips at de + di \u223c 2.5\u2005\u00c5. A low percentage contribution due to O\u22efH/H\u22efO contacts is noted for (II)i.e. 29.1%, contribution from C\u22efH/H\u22efC contacts to the Hirshfeld surfaces of (II)de + di \u223c 2.8\u2005\u00c5 and the distribution of points around de + di \u223c 2.9\u2005\u00c5 in both the wings of its delineated fingerprint plot, Fig.\u20057bi.e. de + di > 3.0\u2005\u00c5, Fig.\u00a07The overall two-dimensional fingerprint plots for (I)R\u2032\u2032xSn(S2CNRR\u2032)x4\u2013, can adopt a variety of coordination geometries, especially for x = 2 2Sn[S2CN(CH3)CH2CH2OCH3]2 . Carbon, hydrogen, nitro\u00adgen and sulfur analyses were performed on a Leco CHNS-932 Elemental Analyzer. The IR spectra were obtained on a Perkin Elmer Spectrum GX from 4000 to 400\u2005cmSynthesis of (I) bis\u00ad(2-meth\u00adoxy\u00adeth\u00adyl)amine dissolved in ethanol (30\u2005ml) was stirred for 30\u2005min. Then, carbon di\u00adsulfide in cold ethanol was added and the resulting mixture was stirred for 2\u2005h. A 25% ammonia solution (1\u20132\u2005ml) was added to generate basic conditions. Then, di-n-butyl\u00adtin(IV) dichloride dissolved in ethanol (20\u201330\u2005ml) was added dropwise into the solution and stirring was continued for 2\u2005h. All reactions were carried out at 277\u2005K. The precipitate that formed was dried and collected. Colourless prisms were harvested from the slow evaporation of its chloro\u00adform:ethanol (2:1 v/v) solution. Yield: 72%. M.p. 341\u2013342\u2005K. Elemental analysis: calculated (%): C 40.68, H 7.14, N 4.31, S 19.75. Found (%): C 41.76, H 6.07, N 4.91, S 19.25. IR (KBr cm\u22121): 1487 \u03bd(C\u2014N), 992 \u03bd(C\u2014S), 544 \u03bd(Sn\u2014C), 386 \u03bd(Sn\u2014S). 1H NMR (CDCl3): \u03b4 4.13 ; 3.70 ; 3.35 ; 1.45\u20132.05 , 0.94 . 13C NMR (CDCl3): \u03b4 201.52 (NCS2); 70.07 (O\u2014CH2); 55.59 (N\u2014CH2); 59.01 (O\u2014CH3); 34.26 Sn\u2014CH2; 28.55 Sn\u2014CH2CH2; 26.41 Sn\u2014CH2CH2CH2; 13.87 CH2CH3. 119Sn NMR (CDCl3): \u03b4 \u2212335.5.Synthesis of (II) The synthesis of (II)v/v) solution. Yield: 78%. M.p. 366-367\u2005K. Elemental analysis: calculated (%): C 53.71, H 4.89, N 2.72, S 12.47. Found (%): C 54.28, H 5.26, N 2.73, S 12.5. IR (KBr cm\u22121): 1477 \u03bd(C\u2014N), 997 \u03bd(C\u2014S), 527 \u03bd(Sn\u2014C), 451 \u03bd(Sn\u2014S). 1H NMR (CDCl3): \u03b4 7.41\u20137.82 ; 4.05 ; 3.71 ; 3.51 ; 3.38 . 13C NMR (CDCl3): \u03b4 196.97 (NCS2); 128.25\u2013142.28 (C-aromatic); 70.09 (O\u2014CH2); 59.06 (N\u2014CH2); 58.10 (O\u2014CH3); 45.81 (N\u2014CH3);. 119Sn NMR (CDCl3): \u03b4 \u2212183.8.Uiso(H) set to 1.2\u20131.5Ueq(C). For (I)\u22123, respectively, were located 0.88 and 1.03\u2005\u00c5 from the S1 and Sn atoms, respectively. For (II)\u22123, respectively, were located 0.96 and 0.76\u2005\u00c5 from the Sn atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S2056989018001901/hb7731sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989018001901/hb7731Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989018001901/hb7731IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1821129, 1821128CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Tumor necrosis factor-\u03b1 has been proven an effective anticancer agent in preclinical studies. However, the translation of TNF\u03b1 from research to clinic has been blocked by significant systemic toxicity and limited efficacy at maximal tolerated dose, which need urgently to be solved.2+ was assessed by Fura-2 in HCC cells. After changing cytosolic Ca2+ level by using agonists or inhibitors, cell apoptosis was detected by flow cytometry. We also detected the effect of ionomycin or parvalbumin on the anti-tumor activity of TNF\u03b1 in a mice model. Lastly, we studied the roles of cytosolic Ca2+ in the mitochondrial-dependent intrinsic apoptosis pathway.The level of cytosolic Ca2+ influx into cytoplasm through transient receptor potential channel in HCC cells. Both cytosolic Ca2+ scavenger and Ca2+-binding protein PV effectively desensitized hepatocellular carcinoma cells to TNF\u03b1, whereas combination ionomycin or 1,4,5-inositol triphosphate significantly sensitized HCC cells to TNF\u03b1, indicating that the increased level of cytosolic Ca2+ was positively correlated with the TNF\u03b1-induced cell apoptosis in vitro. In a nude mice xenograft model, our data revealed that TNF\u03b1 combined with ionomycin remarkably synergized the anti-tumor effect of TNF\u03b1. Furthermore, we found that TNF\u03b1-mediated extracellular Ca2+ influx accelerated TNF\u03b1-induced extrinsic apoptosis through activating calpain/IAP/caspase3 pathway.Here, we demonstrated that TNF\u03b1 induced extracellular Ca2+ influx to enhance cell apoptosis and suggests that increasing the level of cytosolic Ca2+ might be an alternative strategy to improve the pro-apoptotic activity of TNF\u03b1 in HCC cells, although suitable chemical or biological reagents need to be further tested.Our study provides the evidence supporting a novel mechanism by which TNF\u03b1 induces extracellular CaThe online version of this article (10.1186/s13046-018-0714-6) contains supplementary material, which is available to authorized users. TNF\u03b1 is a 23KD type II transmembrane protein, which is arranged in stable homotrimers. It is primarily produced by macrophages and a variety of other cells, including NK cells, T lymphocytes, smooth muscle cells, fibroblasts and others . Many pr2+, as a common signal transduction factor, has played many important roles in the process of cell division, growth, and death c after TNF\u03b1 treatment in HCC cells. We further found that either Ca2+ scavengers or PV reduced the percentage of TNF\u03b1-induced apoptotic HCC cells and IP3 were used to rapidly elevate [Ca2+]c as described previously [2+]c in HLF and QSG-7701 cells after TNF\u03b1 treatment treatment only plays critical roles in TNF\u03b1-induced extrinsic apoptotic pathway c level using fluorescent probe Fura-2/AM in SNU739 and HLF cells with treatment as indicated. BAPTA-AM: 10\u00a0\u03bcM. (b) Cell apoptosis analysis by flow cytometry 24\u00a0h after treatment as indicated before TNF\u03b1 (100\u00a0ng/mL) stimulation. BAPTA-AM: 10\u00a0\u03bcM. (c) Confocal microscope analysis of [Ca2+]c level using fluorescent probe Fura-2/AM in HCC cells with treatment as indicated. EV: cells transfected with the empty vector; PV-OE: cells stably forced expressing Parvalbumin protein. (d) Apoptosis analysis by flow cytometry 24\u00a0h after treatment as indicated. (e) Western Blot analysis for Parvalbumin expression in SNU739 and HLF cells with treatment as indicated. (f) Cell apoptosis analysis by flow cytometry 24\u00a0h after treatment as indicated. CAI: 10\u00a0\u03bcM; SKF96365: 100\u00a0\u03bcM. (g) Cell apoptosis analysis by flow cytometry 24\u00a0h after treatment as indicated. siTRPM7: siRNA against TRPM7; si Ctrl: negative control siRNA. Data were shown as mean\u2009\u00b1\u2009SD. All experiments were performed at least three times. * P\u2009<\u20090.05; ** P\u2009<\u20090.01. (ZIP 1986\u00a0kb)Additional file 4:Figure S3. Cytosolic Ca2+ sensitized HCC cells to TNF\u03b1-induced apoptosis. (a) Apoptosis analysis by flow cytometry 24\u00a0h after treatment as indicated. (b) and (c) Confocal microscope analysis of [Ca2+]c level using fluorescent probe Fura-2/AM in HLF and QSG-7701 cells with treatment as indicated. TNF\u03b1 (50\u00a0ng/mL)\u2009+\u2009Iono: 50\u00a0ng/mL TNF\u03b1 combined with 1\u00a0\u03bcM ionomycin, TNF\u03b1 (100\u00a0ng/mL)\u2009+\u2009Iono: 100\u00a0ng/mL TNF\u03b1 combined with 1\u00a0\u03bcM ionomycin, TNF\u03b1 (200\u00a0ng/mL)\u2009+\u2009Iono: 200\u00a0ng/mL TNF\u03b1 combined with 1\u00a0\u03bcM ionomycin. TNF\u03b1 (50\u00a0ng/mL)\u2009+\u2009IP3: 50\u00a0ng/mL TNF\u03b1 combined with 10\u00a0\u03bcM IP3; TNF\u03b1 (100\u00a0ng/mL)\u2009+\u2009IP3: 100\u00a0ng/mL TNF\u03b1 combined with 10\u00a0\u03bcM IP3; TNF\u03b1 (200\u00a0ng/mL)\u2009+\u2009IP3: 200\u00a0ng/mL TNF\u03b1 combined with 10\u00a0\u03bcM IP3; (d)-(f) Apoptosis analysis by flow cytometry 24\u00a0h after treatment as indicated. Data were shown as mean\u2009\u00b1\u2009SD. All experiments were performed at least three times. * P\u2009<\u20090.05; ** P\u2009<\u20090.01. (ZIP 2448\u00a0kb)Additional file 5:Figure S4. The level of extracellular calcium influx is positively correlated with TNF\u03b1-mediated apoptosis. (a) Confocal microscope analysis of [Ca2+]c level using fluorescent probe Fura-2/AM in 10 kinds of HCC cells with treatment as indicated. (b) Apoptosis analysis by flow cytometry 24\u00a0h after treatment as indicated. All experiments were performed at least three times. (ZIP 2184\u00a0kb)Additional file 6:Figure S5. The role of TNF\u03b1-mediated Ca2+ influx in normal hepatic cells. (a) and (c) Confocal microscope analysis of [Ca2+]c level using fluorescent probe Fura-2/AM in QSG-7701 cells with treatment as indicated. TNF\u03b1: 100\u00a0ng/mL; siTRPM7: siRNA target TRPM7. (b) qRT-PCR and western blot analysis of TRPM7 mRNA and protein expression levels in QSG-7701 cells transfected with siRNA as indicated. (d) Analysis of Calpain activity after TNF\u03b1 stimulation for 4\u00a0h in QSG-7701 cells with treatment as indicated. Data were shown as mean\u2009\u00b1\u2009SD. All experiments were performed at least three times. * P\u2009<\u20090.05; ** P\u2009<\u20090.01. (JPEG 664\u00a0kb)"} +{"text": "A new mononuclear cobalt(III) complex that has two thiol\u00adate and one thio\u00adether donor atoms is reported. I3CoIII2 complex, [Ag3{Co(L)}2]3+ [L3\u2013 = N(CH2NHCH2CH2S\u2212)3], in which two tris\u00ad(thiol\u00adate)-type mononuclear CoIII units ([Co(L)]) are bridged by three AgI ions through S atoms, with iodo\u00admethane (CH3I) gave a new CoIII mononuclear complex, [Co(LMe2)]2+ [LMe2\u2212 = N(CH2NHCH2CH2S\u2212)(CH2NHCH2CH2SCH3)2], systematic name: {2-[(bis{[2-(methylsulfanyl)ethyl]aminomethyl}aminomethyl)amino]ethanethiolato}cobalt(III) bis(hexafluoridophosphate). This cationic complex was crystallized with PF6\u2212 anions to form the title compound, [Co(LMe2)](PF6)2. In the [Co(LMe2)]2+ cation, two of three thiol\u00adate groups in [Co(L)] are methyl\u00adated while one thiol\u00adate group remains unreacted. Although a total of eight stereoisomers are possible for [Co(LMe2)]2+, only a pair of enanti\u00adomers {\u039bRR- and \u0394SS-[Co(LMe2)]2+} are selectively formed. In the crystal, the complex cations and the PF\u00ad6\u2212 anions are connected through weak N\u2014H\u22efF, C\u2014H\u22efF and C\u2014H\u22efS hydrogen bonds into a three-dimensional structure. Two F atoms in one PF6 anion are disordered over two sets of sites with refined occupancies of 0.61\u2005(4) and 0.39\u2005(4) and two F atoms in the other PF6\u2212 anion are disordered over two sets of sites with occupancies of 0.5.Treatment of an S-bridged penta\u00adnuclear Ag R1S\u2212) bound to a transition metal center readily react with alkyl halides (R2X) to form a transition metal complex with thio\u00adether groups (R1SR2). Since the resulting thio\u00adether S atoms generally turn to be asymmetric , the alkyl\u00adated species are an inter\u00adesting research target of coordination stereochemistry. Among a variety of alkyl halides, iodo\u00admethane (CH3I) is one of the most common alkyl\u00adation reagent because of its high reactivity and simple mol\u00adecular structure. For example, the reaction of a mono(thiol\u00adate)-type CoIII mononuclear complex, [Co(aet)(en)2]2+ , with iodo\u00admethane selectively produces the corresponding mono(thio\u00adether)-type complex, [Co(mtea)(en)2]3+ (mtea = NH2CH2CH2SCH3) 2], is also easily converted to the corres\u00adponding bis\u00ad(thio\u00adether)-type complex, [Ni(mtea)2]2+, by treating with iodo\u00admethane. Unlike mono(thiol\u00adate)- or bis\u00ad(thiol\u00adate)-type complexes, tris\u00ad(thiol\u00adate)-type complexes have been found to show different reactivity toward iodo\u00admethane. That is, the reaction of a tris\u00ad(thiol\u00adate)-type mononuclear rhodium(III) complex, fac(S)-[Rh(aet)3], with iodo\u00admethane afforded a unique di\u00admethyl\u00adated mono(thiol\u00adate)bis\u00ad(thio\u00adether)-type complex, fac(S)-[Rh(aet)(mtea)2]2+, whereas the mono\u00admethyl\u00adated bis\u00ad(thiol\u00adate)mono(thio\u00adether)-type and tri\u00admethyl\u00adated tris\u00ad(thio\u00adether)-type species were little formed -[Rh(aet)(mtea)2]2+. However, the lack of crystallographic analytical data for fac(S)-[Rh(aet)(mtea)2]2+ prevented the further study on the stereochemistry of the di\u00adalkyl\u00adated complex.It has long been recognized that thiol\u00adate groups ]2}3+ [L3\u2013 = N(CH2NHCH2CH2S\u2212)3] ], are linearly linked by three AgI ions, reacts with iodo\u00admethane to give a mono(thiol\u00adate)bis\u00ad(thio\u00adether)-type complex, [Co(LMe2)]2+ [LMe2\u2212 = N(CH2NHCH2CH2S\u2212)(CH2NHCH2CH2SCH3)2]. It is noteworthy that the complex was crystallized as a hexa\u00adfluorido\u00adphosphate salt, [Co(LMe2)](PF6)2, and its mol\u00adecular structure was fully determined by single-crystal X-ray diffraction analysis. As far as we know, this is the first crystallographic characterization of a cobalt(III) complex that has two thio\u00adether and one thiol\u00adate donor groups. In addition, this is a unique example of a direct conversion of a thiol\u00adate-bridged multinuclear complex to a mononuclear thio\u00adether complex by alkyl\u00adation reaction. Treatment of the thiol\u00adate-bridged penta\u00adnuclear complex {Ag3[Co(L)]2}3+ with excess iodo\u00admethane in water gave a greenish-brown suspension. After removing the insoluble solid by filtration, the purple\u2013brown filtrate was purified by a cation\u2013exchange column (SP-Sephadex C-25). The product was isolated as purple\u2013brown crystals by adding a hexa\u00adfluorido\u00adphosphate anion. The geometrical parameters and stereoisomerism of the title compound based on the X-ray analysis, together with the spectroscopic data, are described in this paper.In the course of our continuing study of the alkyl\u00adation reaction of metal complexes with amino\u00adthiol\u00adate ligands (Okamoto LMe2)]2+, and four PF6\u2212 anions in the asymmetric unit ] moiety were methyl\u00adated to form [Co(LMe2)]2+. No apparent difference was observed among the Co\u2014S bond lengths for thiol\u00adate S atoms (Sthiol\u00adate) [2.2384\u2005(13)\u20132.2478\u2005(11)\u2005\u00c5] and those for thio\u00adether S atoms (Sthio\u00adether) [2.2190\u2005(13)\u20132.2599\u2005(11)\u2005\u00c5] in [Co(LMe2)]2+. However, the Co\u2014N bonds trans to Sthiol\u00adate [2.061\u2005(4)\u20132.062\u2005(3)\u2005\u00c5] are ca 0.05\u2005\u00c5 longer than the Co\u2014N bonds trans to Sthio\u00adether [2.004\u2005(4)\u20132.020\u2005(4)\u2005\u00c5]. The difference is reasonably explained by the decrease of the trans influence due to the alkyl\u00adation on S atoms. As a result of the steric repulsion between the methyl groups on the S atoms, the S\u2014Co\u2014S angles in [Co(LMe2)]2+ deviate considerably from 90\u00b0 [86.58\u2005(4)\u201395.07\u2005(4)\u00b0].X-ray structural analysis revealed that there are two crystallographically independent yet essentially the same complex cations, units. Considering the absolute configurations of the cobalt(III) atom (\u0394 and \u039b) and the two asymmetric sulfur atoms (R and S), four pairs of diastereomers, \u0394SS/\u039bRR, \u0394SR/\u039bRS, \u0394RS/\u039bSR and \u0394RR/\u039bSS, are possible for [Co(LMe2)]2+. However, the asymmetric unit of this crystal contains two \u039bRR isomers. As indicated by the space group C2/c, the title crystal is a racemic compound consisting of a pair of enanti\u00adomers, \u039bRR and \u0394SS. This result is consistent with the observation that the 13C{1H} NMR spectrum of the title compound in DMSO-d6 exhibits a total of 10 sharp singlet signals, assignable to the C1 symmetrical \u039bRR and \u0394SS isomers of [Co(LMe2)]2+ ]2+ in the crystal, two of three N,S-chelate rings have a gauche form with the lel (\u03bb for \u0394 and \u03b4 for \u039b) conformation, while one has a gauche form with the ob (\u03bb for \u039b and \u03b4 for \u0394) conformation.Each Co2+ Fig.\u00a02. For botLMe2)]2+. This complex was obtained by the unprecedented direct conversion of a thiol\u00adate-bridged AgI3CoIII2 penta\u00adnuclear complex by alkyl\u00adation reaction using iodo\u00admethane. The selective formation of the \u039bRR and \u0394SS isomers of [Co(LMe2)]2+ observed in the crystal structure is consistent with the result of 13C{1H} NMR. The findings reported herein will provide insight into the synthesis and structures of coordination compounds containing both thiol\u00adate and thio\u00adether donor groups.In summary, we report here the first example of a crystallographically characterized mono(thiol\u00adate)bis\u00ad(thio\u00adether)-type mononuclear cobalt(III) complex, [Co]2}(NO3)3\u00b74H2O in 100\u2005ml of water was added CH3I . The mixture was stirred at room temperature for 1.5 days in the dark. After removing a brown powder (200\u2005mg) by filtration, the purple\u2013brown filtrate was poured onto an SP-Sephadex C-25 column . First, a purple band was eluted with 0.05\u2005M aqueous NaCl. Then, a purple\u2013brown band of [Co(LMe2)]2+ was eluted with 0.15\u2005M aqueous NaCl. To the concentrated purple\u2013brown eluate was added 1.0\u2005M aqueous NH4PF6 (5\u2005ml) and the solution was allowed to stand at room temperature for 20\u2005d. The resulting dark purple\u2013brown block crystals of the title compound were collected by filtration. Yield: 0.08\u2005g (29%). Single crystals suitable for X-ray analysis were obtained by recrystallization from water by adding 1.0 M aqueous NH4PF6. Analysis: calculated for [Co(LMe2)](PF6)2: C 20.01, H 4.12, N 8.48%; found: C 20.25, H 4.06, N 8.51%. 13C{1H} NMR (DMSO-d6): \u03b4 17.40, 18.05, 28.97, 37.20, 47.82, 49.42, 58.22, 64.39, 67.05, 67.50. One of the 13C signals overlaps with the signal from solvent. IR: 3266.8(m), 3029.6(w), 1432.8(m), 1245.8(w), 1158.0(w), 1113.7(w), 1034.6(w), 955.5(m), 839.8(s), 558.3(s).To a dark-purple solution of {Ag2) or 0.98\u2005\u00c5 (CH3)] and refined as riding with Uiso(H) = 1.2Ueq(C) for CH2 and Uiso(H) = 1.5Ueq(C) for CH3. All H atoms bound to N atoms were refined with bond-length restraints [N\u2014H = 0.90\u2005(2)\u2005\u00c5] and with Uiso(H) = 1.2Ueq(N). Two F atoms in one PF6 anion are disordered over two positions (F25A/F25B and F26A/F26B) with refined occupancies of 0.61\u2005(4) and 0.39\u2005(4). Two F atoms in another PF6 anion are also disordered over two positions with site occupancies of 0.5. Reflections I. DOI: 10.1107/S2056989017005229/is5473Isup2.hklStructure factors: contains datablock(s) I. DOI: 1542478CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The 1,3-di\u00admethyl\u00adurea mol\u00adecules are also hydrogen bonded to sulfate O atoms, and project outwards from the ribbon parallel to the b axis.The packing is centred on bis\u00ad(piperidinium) sulfate ribbons parallel to the 5H12N+\u00b7SO42\u2212\u00b72C3H8N2S, the C=S groups of the two independent 1,3-di\u00admethyl\u00adurea mol\u00adecules and the sulfur atom of the anion lie on twofold axes. The packing is centred on bis\u00ad(piperidinium) sulfate ribbons parallel to the c axis; the cations are hydrogen bonded to the sulfate by N\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions. The 1,3-di\u00admethyl\u00adurea mol\u00adecules are also hydrogen bonded to sulfate O atoms, and project outwards from the ribbon parallel to the b axis.In the title compound, 2C We have published two reports on adducts of dioxane and morpholine with various methyl\u00adthio\u00adureas , so that the methyl groups are cis, with Cmeth\u00adyl\u2014N\u2014C=S torsion angles are close to zero [C11\u2014N1\u2014C1\u2014S1 = 0.9\u2005(2)\u00b0 and S2\u2014C2\u2014N2\u2014C21 = \u22122.05\u2005(17)\u00b0]. Free 1,3-DMT crystallizes with four independent mol\u00adecules, each of which has one NH group cis and one trans to C=S, but the structure is severely disordered sulfate and 1,3-DMT Fig.\u00a01, with thc axis in the region x, y \u2243 1/2 sulfate ribbons parallel to the /2 Fig.\u00a02 and also2 Table\u00a01. Each pa2 Table\u00a01 and 3 \u25b8;et al., 2016trans geometry for both NH functions, but the 1:2 adduct between 1,4-dioxane and 1,3-DMT hydrogensulfate sulfate 1,3-DMT were dissolved in 2\u2005mL piperidine. The solution was overlayered with diethyl ether. Colourless needles formed overnight.y, Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017004820/hg5484sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017004820/hg5484Isup2.hklStructure factors: contains datablock(s) I. DOI: 1540583CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The aryl\u00adoxo magnesiate complex anion is binuclear with each Mg2O4 complex unit inversion-related and bridged through the two tridentate chelating phenolate O-donors of the ligand. The complex centres have a distorted tetra\u00adhedral stereochemistry [Mg\u2014O range 1.8796\u2005(17)\u20132.0005\u2005(16)\u2005\u00c5] and an Mg\u22efMg separation of 2.9430\u2005(14)\u2005\u00c5]. The LiO4 coodination sphere of the cation comprises four THF O-donor atoms and has a slightly distorted tetra\u00adhedral conformation [Li\u2014O range 1.899\u2005(5)\u2013 1.953\u2005(5)\u2005\u00c5]. In the crystal, a number of stabilizing intra-anion C\u2014H\u22efO hydrogen-bonding inter\u00adactions are present but no inter-species associations are found.The title ion-association metal complex, [Li(C The structure of this novel heterobimetallic complex, in an ion-association mode, is reported herein.Magnesium complexes of the two chelating phenolate groups . The stereochemistry about each four-coordinated Mg atom is distorted tetra\u00adhedral with Mg\u2014O(Ar) in the range 1.8796\u2005(17)\u20132.0005\u2005(16)\u2005\u00c5 \u2013 1.953\u2005(5)\u2005\u00c5. Within the binuclear complex anion there are six stabilizing intra-ion methyl C\u2014H\u22efO hydrogen-bonding inter\u00adactions methane and nBuLi was stirred in THF (20\u2005mL) at 273\u2005K under an N2 atmosphere for 2\u2005h. MgEt2 was gently added to the solution. After stirring at 298\u2005K for 6\u2005h, the solution was filtered through celite. The filtrate was concentrated to ca 10\u2005mL and cooled to 273\u2005K to furnish colourless crystals, suitable for the X-ray analysis. Yield: 0.46\u2005g (49%).A solution of tris\u00ad, or 1.5Ueq(methyl C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017008337/zs2380sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017008337/zs2380Isup2.hklStructure factors: contains datablock(s) I. DOI: 1554461CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-pyrazol-5(4H)-one and its structural characterization by synchrotron single-crystal X-ray diffraction are reported.A synthetic approach to the new zinc complex based on amino\u00admethyl\u00adene derivative of 1-phenyl-3-methyl-4-[(quinolyl-3-yl)imino\u00admeth\u00adyl]-1 20H15N4O)2]\u00b72.5CH3OH, I, was synthesized via the reaction of zinc acetate with the respective ligand and isolated as a methanol solvate, i.e., as 3OHI\u00b72.5CH. The crystal structure is triclinic (space group P-1), with two complex mol\u00adecules (A and B) and five methanol solvent mol\u00adecules in the asymmetric unit. One of the five methanol solvent mol\u00adecules is disordered over two sets of sites, with an occupancy ratio of 0.75:0.25. Mol\u00adecules A and B are conformers and distinguished by the conformations of the bidentate 1-phenyl-3-methyl-4-[(quinolin-3-yl)imino\u00admeth\u00adyl]-1H-pyrazol-5-olate ligands. In both mol\u00adecules, the zinc cations have distorted tetra\u00adhedral coordination spheres, binding the monoanionic ligands through the pyrazolo\u00adlate O and imine N atoms. The two ligands adopt slightly different conformations in terms of the orientation of the terminal phenyl and quinoline substituents with respect to the central pyrazolo\u00adlate moiety. The mol\u00adecular geometries of A and B are supported by intra\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds. In the crystal of I, mol\u00adecules form dimers both by secondary inter\u00admolecular Zn\u22efO [3.140\u2005(2)\u20133.553\u2005(3)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking inter\u00adactions. The dimers are linked by inter\u00admolecular hydrogen bonds through the solvent methanol mol\u00adecules into a three-dimensional network.The title compound, [Zn(C ComplexA and B in I are four-coordinated by two monoanionic O,N-chelating ligands, which bind to the cation through pyrazolo\u00adlate O and imine N atoms. The coordination sphere around each zinc cation can be described as distorted tetra\u00adhedral , with dihedral angles between the planar six-membered chelating rings of 82.97\u2005(7) and 84.52\u2005(7)\u00b0 for mol\u00adecules A and B, respectively.The zinc cations of A and B of I adopt different conformations. The main difference pertains to the twist angles of the terminal phenyl and quinoline substituents relative to the central imino\u00admethyl-1H-pyrazol-5-olate fragment. In mol\u00adecule A and 8.24\u2005(11)\u00b0 for the phenyl groups, and 27.47\u2005(10) and 26.08\u2005(6)\u00b0 for the quinoline substituents. Thus, one of the two 1-phenyl-3-methyl-4-[(quinolin-3-yl)imino\u00admeth\u00adyl]-1H-pyrazol-5-olate ligands in mol\u00adecule B is flattened, while one of the two pyrazolo\u00adlate ligands in mol\u00adecule A is substanti\u00adally twisted \u2005\u00c5, Zn1\u22efO4 = 3.279\u2005(3)\u2005\u00c5, Zn2\u22efO1 = 3.553\u2005(3)\u2005\u00c5 and Zn2\u22efO2 = 3.140\u2005(2)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking inter\u00adactions between the O2/N5/N6/N7/C21\u2013C24 and O4/N13/N14/N15/C61\u2013C64 imino-methyl-pyrazolo\u00adnate fragments {the shortest distances are N6\u22efC63 [3.083\u2005(3)\u2005\u00c5], C21\u22efC62 [3.210\u2005(4)\u2005\u00c5], C24\u22efC64 [3.216\u2005(4)\u2005\u00c5], C21\u22efC61 [3.261\u2005(3)\u2005\u00c5], N14\u22efC23 [3.293\u2005(4)\u2005\u00c5], C22\u22efC61 [3.297\u2005(4)\u2005\u00c5], N6\u22efC62 [3.319\u2005(3)\u2005\u00c5] and N14\u22efC22 [3.362\u2005(3)\u2005\u00c5]}, as well as phenyl and pyridine rings , where Cg3vii is the centroid of the C69vii\u2013C74vii benzene ring; symmetry code: (vii) \u2212x, \u2212y, \u2212z\u00a0+\u00a01] and \u03c0\u2013\u03c0 stacking inter\u00adactions {the shortest distances are between the C75\u2013C80 and C75viii\u2013C80viii phenyl rings [C75\u22efC79viii = 3.196\u2005(4)\u2005\u00c5 and C80\u22efC80viii = 3.279\u2005(4)\u2005\u00c5]; symmetry code: (viii) \u2212x, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01}, as well as C\u2014H\u22efO and N\u22efH\u2014O hydrogen bonds involving the solvent methanol mol\u00adecules : 1664 \u03bd(C=O), 1627 \u03b4(NH). 1H NMR : \u03b4 2.31 , 7.08\u20138.03 , 8.52 , 8.89 , 11.46 . UV\u2013vis spectrum (nm): 232, 254, 358. PL spectrum (nm): \u03bbPL = 454, 534, \u03bbex = 450\u2005nm. Quantum yield of PL \u03c6 = 0.002. Analysis calculated for C20H16N4O: C 73.15, H 4.91, N 17.06%; found: C 73.25, H 5.10, N 17.18%.A solution containing 1.44\u2005g (0.01\u2005mol) of 3-amino\u00adquinoline in 10\u2005ml of toluene was added to a solution of 2.02\u2005g (0.01\u2005mol) of 1-phenyl-3-methyl-4-formyl\u00adpyrazol-5-one in 20\u2005ml of toluene. The mixture was refluxed for 3\u2005h with a Dean\u2013Stark trap until water stripping was completed. Subsequently, two-thirds of the total volume was distilled off on a rotary evaporator. The precipitate which formed was filtered off and recrystallized from ethanol to give light-yellow crystals . FT\u2013IR in KBr in 20\u2005ml of the same solvent mixture and dried at 423\u2005K, resulting in a yellow crystalline powder . FT\u2013IR : 1608 \u03bd(C=N). 1H NMR : \u03b4 2.25 , 6.99\u20138.92 , 8.46 . UV\u2013vis (nm): 360, 340, 304. PL (nm): \u03bbPL = 478, \u03bbex = 450\u2005nm. Analysis calculated for C40H30N8O2Zn: C 66.72, H 4.20, N 15.56%; found: C 66.78, H 4.25, N 15.64, Zn 9.11%.A hot solution of 0.22\u2005g of zinc acetate dihydrate (1\u2005mmol) in 20\u2005ml of methanol was added to hot solutions of nt Fig.\u00a05. The reaCrystal data, data collection and structure refinement details are summarized in Table\u00a02Scala program using a Rayonix SX165 CCD detector. A total of 360 images were collected using an oscillation range of 1.0\u00b0 and corrected for absorption using the The data completeness of 97.8% is caused by the low (triclinic) crystal symmetry. It is very difficult to get a high data completeness for this symmetry using the \u03c6 scan mode only (\u2018Belok\u2019 beamline limitation), even though we have run two different crystal orientations.I/\u03c3 statistics for high-angle reflections, we selected an exposure time so as to admit a minor fraction of intensity overloads in the low-angle part of the detector. These low-angle reflections have imprecisely measured intensities and thus were excluded from the final steps of refinement. (ii) In the present set-up of the synchrotron diffractometer, the low-temperature device eclipses a small region of the 2D detector near the high-angle limit. This small shadowed region has not been masked during integration of the diffraction frames, which erroneously resulted in zero intensity of some reflections. (iii) The quality of the single crystal chosen for the diffraction experiment was not perfect. Some systematic differences between the calculated and observed intensities are probably caused by extinction and defects present in the crystal specimen.A rather large number of reflections have been omitted from refinement due to the following reasons. (i) In order to achieve better Uiso(H) = 1.5Ueq(O)]. All other H atoms were placed in calculated positions, with C\u2014H = 0.95\u20130.98\u2005\u00c5, and refined in a riding mode, with fixed isotropic displacement parameters [Uiso(H) = 1.5Ueq(C) for the CH3 groups and 1.2Ueq(C) for the other groups]. Disorder over two sets of sites was observed for one methanol solvent mol\u00adecule (atoms O7\u2013C83). In the last cycles of refinement, the occupancy ratio was fixed at 0.75:0.25 and each of the non-H atoms was modelled with a common displacement ellipsoid.The H atoms of the hy\u00addroxy groups were localized from difference Fourier maps and included in a riding mode, with fixed displacement parameters [10.1107/S2056989017010441/wm5401sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989017010441/wm5401Isup2.hklStructure factors: contains datablock(s) I. DOI: 1562057CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-(4-chloro\u00adbenzene\u00adsulfon\u00adyl)glycinyl hydrazide with 4-nitro\u00adbenzaldehyde gives the N,N-di\u00admethyl\u00adformamide monosolvated N-acyl\u00adhydrazone derivative, (E)-N-{2-[2-(4-nitro\u00adbenzyl\u00adidene)- hydrazine-1-yl]-2-oxoeth\u00adyl}-4-\u03c7hloro\u00adbenzene\u00adsulfonamide. Rings of Reaction of 15H13ClN4O5S\u00b7C3H7NO, contains one mol\u00adecule each of the Schiff base and the solvent di\u00admethyl\u00adformamide. The hydrazone group adopts an E configuration about the C=N bond. The dihedral angle between the two aromatic rings is 86.58\u2005(2)\u00b0. In the crystal, pairs of N\u2014H\u22efO hydrogen bonds between centrosymmetrically related mol\u00adecules generates rings with an R22(10) graph-set motif. The dimers are further linked via N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, leading to the formation of R33(11) ring motifs. C\u2014H\u22ef\u03c0 inter\u00adactions are also observed. The inter\u00admolecular inter\u00adactions in the crystal structure were qu\u00adanti\u00adfied and analysed using Hirshfeld surface analysis, which indicates that the most significant contacts in packing are O\u22efH/H\u22efO (31.3%), followed by H\u22efH (25.4%) and C\u22efH/H\u22efC (13.0%).The asymmetric unit of the title compound, C The acidity of the C\u2014H donor group determines the strength of C\u2014H\u22efO inter\u00adactions -4-chloro-N-{2-[2-(4-nitro\u00adbenzyl\u00adidene)hydrazin-1-yl]-2-oxoeth\u00adyl}benzene\u00adsulfonamide N,N-di\u00admethyl\u00adformamide monosolvate.Supra\u00admolecular chemistry is based upon non-covalent inter\u00adactions such as hydrogen bonding, \u03c0\u2013\u03c0 stacking and van der Waals inter\u00adactions and 1.274\u2005(6)\u2005\u00c5, respectively, confirm their double-bond character. The C8\u2014N2 and N2\u2014N3 bond distances are shorter than normal bond lengths as a result of delocalization of the \u03c0-electron density. The mol\u00adecule is twisted at N1\u2014C7 with an S1\u2014N1\u2014C7\u2014C8 torsion angle of 166.5\u2005(4)\u00b0. The other central part of the mol\u00adecule is almost linear with C7\u2014C8\u2014N2\u2014N3, C8\u2014N2\u2014N3\u2014C9 and N2\u2014N3\u2014C9\u2014C10 torsion angles of \u22121.6\u2005(7), \u2212179.7\u2005(5) and 177.9\u2005(4)\u00b0, respectively. The orientations of the sulfonamide group with respect to the attached phenyl ring is given by the torsion angles of C2\u2014C1\u2014S1\u2014N1 = 98.1\u2005(5)\u00b0 and C6\u2014C1\u2014S1\u2014N1 = \u221280.2\u2005(5)\u00b0, while that of the hydrazone group with the attached phenyl ring by the torsion angles of C11\u2014C10\u2014C9\u2014N3 = 1.6\u2005(8)\u00b0 and C15\u2014C10\u2014C9\u2014N3 = \u2212177.4\u2005(5)\u00b0. The dihedral angle between the sulfonyl benzene ring and the mean plane through the SO2\u2014NH\u2014CH2\u2014CO segment is 82.653\u2005(18)\u00b0, while that between the C10\u2013C15 phenyl ring and the mean plane through the C9\u2014N3\u2014N2\u2014CO segment is 4.44\u2005(3)\u00b0. The dihedral angle between the two aromatic rings is 86.58\u2005(2)\u00b0. The C1\u2013C6 and C10\u2013C15 benzene rings are inclined to the mean plane of the central part of the hydrazone mol\u00adecule by 86.4\u2005(3) and 4.5\u2005(3)\u00b0, respectively.The asymmetric unit of the title compound Fig.\u00a01 containsvia N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, generating rings with an via N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, leading to the formation of B) and the sulfonyl oxygen atom (O2) forming c axis, and the other involving an aromatic C\u2014H (H14) and the nitro O4 atom, giving rise to inversion dimers with an E)-4-methyl-N-{2-[2-(4-nitro\u00adbenz\u00adyl\u00adidene)hydrazin-1-yl]-2-oxoeth\u00adyl}benzene\u00adsulfonamide N,N-di\u00admeth\u00adyl\u00adform\u00adamide monosolvate to analyse the close contacts in the title compound. The electrostatic potentials were calculated using TONTO surface resolution with the 3D dnorm surfaces mapped over a fixed colour scale of \u22120.5849 to 1.3948. The curvedness was mapped in the colour range of \u22124.0 to 0.4. The electrostatic potentials were mapped on Hirshfeld surfaces using the STO-3G basis set at the Hartree\u2013Fock level theory over a range \u00b10.1au.dnorm . The H\u22efH inter\u00adactions appear as the largest region of the fingerprint plot with a high concentration in the middle region, shown in light blue, at de = di \u223c1.4\u2005\u00c5 with an overall contribution to the Hirshfeld surfaces of 25.4%. The C\u22efH contacts, which refer to C\u2014H\u22ef\u03c0 inter\u00adactions, contribute 13.0% of the Hirshfeld surfaces. The presence of C\u2014H\u22ef\u03c0 inter\u00adactions is indicated by the appearance of two broad spikes having almost same de + di 3.1\u2005\u00c5. The C\u22efC contacts contribute 4.5% of the Hirshfeld surfaces, featuring two successive triangles with a minimum (de + di) distance of \u223c3.5\u2005\u00c5, which is greater than van der Waals separation, confirming the absence of \u03c0\u2013\u03c0 stacking inter\u00adactions. This is also evident from the absence of flat regions in the Hirshfeld surface mapped over curvedness glycine (L1) obtained was crystallized from aqueous ethanol. Sulfuric acid (0.5\u2005ml) was added to L1 (0.02\u2005mol) dissolved in ethanol (30\u2005ml) and the mixture was refluxed. The reaction mixture was monitored by TLC at regular inter\u00advals. After completion of the reaction, the reaction mixture was concentrated to remove the excess ethanol. The product, N-(4-chloro\u00adbenzene\u00adsulfon\u00adyl)glycine ethyl ester (L2) obtained was poured into water, neutralized with sodium bicarbonate and recrystallized from acetone. The pure L2 (0.01\u2005mol) was then added in small portions to a stirred solution of 99% hydrazine hydrate (10\u2005ml) in 30\u2005ml ethanol and the mixture was refluxed for 6\u2005h. After cooling to room temperature, the resulting precipitate was filtered, washed with cold water and dried to obtain N-(4-chloro\u00adbenzene\u00adsulfon\u00adyl)glycinyl hydrazide (L3). A mixture of L3 (0.01\u2005mol) and 4-nitro\u00adbenzaldehyde (0.01\u2005mol) in anhydrous methanol (30\u2005ml) and two drops of glacial acetic acid was refluxed for 8h. After cooling, the precipitate was collected by vacuum filtration, washed with cold methanol and dried. It was recrystallized to a constant melting point from methanol (493\u2013496\u2005K).4-Chloro\u00adbenzene\u00adsulfonyl chloride (0.01\u2005mol) was added to glycine (0.02\u2005mol) dissolved in an aqueous solution of potassium carbonate . The reaction mixture was stirred at 373\u2005K for 6\u2005h, left overnight at room temperature, then filtered and treated with dilute hydro\u00adchloric acid. The solid \u22121 for the stretching bands of N\u2014H, C=O, C=N, S=O asymmetric and S=O symmetric, respectively. 1H NMR : 3.68, 4.17 , 7.62\u20137.67 , 7.80\u20137.94 , 8.24\u20138.29 , 8.02 , 8.14 , 11.73, 11.75 . 13C NMR : 43.26, 44.42, 123.94, 127.85, 128.53, 129.19, 137.23, 139.77, 141.47, 144.68, 147.75, 164.52, 169.34. Plate-like yellow single crystals of the title compound suitable for X-ray analysis were grown from its DMF solution by slow evaporation of the solvent.The purity of the compound was checked by TLC and characterized by its IR spectrum. The characteristic absorptions observed are 3250.1, 1685.8, 1587.4, 1342.5 and 1166.9\u2005cmUeq or 1.5Ueq(C) for methyl H atoms.. The amino H atoms were freely refined with the N\u2014H distances restrained to 0.86\u2005(2)\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901800292X/rz5227sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901800292X/rz5227Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901800292X/rz5227Isup3.cmlSupporting information file. DOI: 1433601CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The metal atom of the title compound is four coordinated. The asymmetrically appended Cl atom and a widely spread \u03c0-conjugated system of the complex mol\u00adecule construct the supra\u00admolecular structures of a hydrogen-bonded chain and a \u03c0\u2013\u03c0 inter\u00adacted column. 20H14ClN3O)], with an asymmetrically chloride-appended Schiff base ligand has been synthesized and structurally characterized at 100\u2005K. In the compound, the central nickel(II) ion has a square-planar coordination geometry with N3O donors of the \u03c0-conjugated tetra\u00addentate Schiff base ligand. In the crystal, the complexes are connected into an inversion dimer via an Ni\u22efNi inter\u00adaction [3.1753\u2005(5)\u2005\u00c5] and a pair of \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.8416\u2005(16)\u2005\u00c5]. The dimers are linked via a C\u2014H\u22efCl hydrogen bond, forming a chain along the c-axis direction. The dimer chains inter\u00adact with each other through \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.8736\u2005(16)\u2005\u00c5], forming a layer expanding parallel to the ac plane.The title complex, [Ni(C Up to now, a large number of salen derivatives have been prepared and used for complexation in the expectation of a wide range of features such as catalytic ability, magnetic, dielectric and luminescence properties and so on complex using an asymmetrically chloride-appended tetra\u00addentate Schiff base ligand.Metal complexes with a tetra\u00addentate Schiff base ligand as represented by H3O type asymmetrical ligand was reported by Ghorai & Mukherjee complex with a similar Nrjee 2014.3O donor set including one phenolate O atom, two imine N atoms and one amino N atom of the tetra\u00addentate Schiff base ligand atom is in a square-planar coordination with an asymmetrical coordination environment formed by the Nnd Fig.\u00a01. The Ni\u2014et al., 1996c axis \u2005\u00c5] and a pair of \u03c0\u2013\u03c0 inter\u00adactions between the C1\u2013C6 and C15\u2013C20 benzene rings [centroid\u2013centroid distance = 3.8415\u2005(16)\u2005\u00c5]. Such dimerization caused by an Ni\u22efNi inter\u00adaction has also been observed in symmetric Ni compounds of Ni(CH3COO)2\u00b74H2O was added to the solution and stirred for 1\u2005h. The resulting solution was allowed to stand for a few days, during which time dark-purple block-shaped crystals precipitated. They were collected by suction filtration and dried in air to give single crystals of the title compound suitable for X-ray diffraction.The tetra\u00addentate Schiff base ligand was prepared by the reaction of 2-amino\u00adbenzaldehyde = 1.5Ueq(N). Other H atoms were treated as riding with C\u2014H = 0.95\u2005\u00c5 and Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017004613/is5472sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989017004613/is5472Isup2.hklStructure factors: contains datablock(s) I. DOI: 1539785CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular and crystal structure of the insecticide fipronil is reported. In the crystal, N\u2014H\u22efN, N\u2014H\u22efO, C\u2014H\u22efF hydrogen bonds, C\u2014N\u22ef\u03c0 and C\u2014Cl\u22ef\u03c0 inter\u00adactions link adjacent mol\u00adecules, forming a three-dimensional network. In addition, there are short F\u22efF inter\u00adactions present. 12H4Cl2F6N4OS {systematic name: 5-amino-1--4-[(tri\u00adfluoro\u00admethane)sulfinyl]-1H-pyrazole-3-carbo\u00adnitrile}, is a member of the phenyl\u00adpyrazole group of acaricides, and one of the phenyl\u00adpyrazole group of insecticides. The dihedral angle between the planes of the pyrazole and benzene rings is 89.03\u2005(9)\u00b0. The fluorine atoms of the tri\u00adfluoro\u00admethyl substituent on the benzene ring are disordered over two sets of sites, with occupancy ratios 0.620\u2005(15):0.380\u2005(15). In the crystal, C\u2014N\u22ef\u03c0 inter\u00adactions [N\u22efring centroid = 3.607\u2005(4)\u2005\u00c5] together with N\u2014H\u22efN and C\u2014H\u22efF hydrogen bonds form a looped chain structure along [10The title compound, C It is an insecticide with extended use in the control of many agricultural vermin. Fipronil contains a tri\u00adfluoro\u00admethyl\u00adsulfinyl substituent that is not present in any other agrochemicals and this is thought to contribute to its remarkable potency in the field \u2005\u00c5; Cg1 is the centroid of the C5\u2013C10 ring; symmetry code: (iii) \u2212x, y, \u2212z], together with N3\u2014H3A\u22efN4i and C9\u2014H9\u22efF2i hydrogen bonds, forming looped chains along (red dashed lines), link adjacent chains, resulting in a two-dimensional network parallel to the (10B\u22efO1ii hydrogen bonds (black dashed lines) combine with these contacts to generate a three-dimensional network structure \u2005\u00c5] and F3\u22efF6\u2032vi [2.855\u2005(12)\u2005\u00c5] inter\u00adactions are also present .In the crystal, mol\u00adecules are linked by C12\u2014N4\u22ef0] Fig.\u00a02. Inversine Fig.\u00a03. Finallyre Fig.\u00a04. Short Fet al., 2005et al., 2013II, CdII, ZnII and MnII complexes using fipronil as a ligand are known -methyl\u00adidene]amino}-1,5-dimethyl-2-phenyl-1H-pyrazol-3(2H)-one = 0.88\u2005\u00c5, Uiso = 1.2Ueq(C) for the N\u2014H group, d(C\u2014H) = 0.95\u2005\u00c5, Uiso = 1.2Ueq(C) for aromatic C\u2014H. Atoms F4\u2013F6 of the CF3 substituent are disordered over two sets of sites. Their occupancies refined to 0.620\u2005(15) and 0.380\u2005(15).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901701310X/sj5534sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S205698901701310X/sj5534Isup2.hklStructure factors: contains datablock(s) I. DOI: 1574261CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of the title compound consists of discrete octa\u00adhedral complexes that are linked by inter\u00admolecular O\u2014H\u22efS, C\u2014H\u22efCl, C\u2014H\u22efS and C\u2014H\u22efCl hydrogen bonding. 2(C5H4ClN)2(H2O)2], consists of one nickel(II) cation that is located on a center of inversion and one thio\u00adcyanate anion, one water mol\u00adecule and one 2-chloro\u00adpyridine ligand all occupying general positions. The NiII cation is octa\u00adhedrally coordinated by two terminal N-bound thio\u00adcyanato ligands, two aqua ligands and two N-bound 2-chloro\u00adpyridine ligands into discrete complexes. Individual complexes are linked by inter\u00admolecular O\u2014H\u22efS and O\u2014H\u22efCl hydrogen-bonding inter\u00adactions into a layered network extending parallel to the bc plane. Weak inter\u00adactions of types C\u2014H\u22efS and C\u2014H\u22efCl consolidate the crystal packing.The asymmetric unit of the title compound, [Ni(NCS) The cation is located on a center of inversion whereas all ligands are located on general positions. The NiII cation is coordinated by two terminal N-bound inorganic anionic ligands, two water mol\u00adecules and two 2-chloro\u00adpyridine ligands that are coordinated via the pyridine N atom in an all-trans configuration on Fig.\u00a01. As expeb axis \u00b76H2O and 2-chloro\u00adpyridine were purchased from Alfa Aesar. Ni(NCS)2 was synthesized by stirring 17.5\u2005g Ba(NCS)2\u00b73H2O (57\u2005mmol) with 15.0\u2005g Ni(SO4)\u00b76H2O (57\u2005mmol) in 500\u2005ml water. The green residue was filtered off and the filtrate was dried using a rotary evaporator. The homogeneity was checked by X-ray powder diffraction and elemental analysis. Crystals of the title compound suitable for single crystal X-ray diffraction were obtained by the reaction of 26.2\u2005mg Ni(NCS)2 (0.15\u2005mmol) with 56.0\u2005\u00b5l 2-chloro\u00adpyridine (0.6\u2005mmol) in ethanol (1.0\u2005ml) after a few days.Ba(NCS)Uiso(H) = 1.2Ueq(C). The OH H atoms were located in a difference map, and their bond lengths constrained to 0.82\u2005\u00c5, with Uiso(H) = 1.5Ueq(O).Crystal data, data collection, and structure refinement details are summarized in Table\u00a0210.1107/S2056989016015218/wm5326sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016015218/wm5326Isup2.hklStructure factors: contains datablock(s) I. DOI: 1506903CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "One of the two n-pentyl side chains was refined as disordered over two sets of sites, with occupancies of 0.733\u2005(18) and 0.267\u2005(18). The geometry around the HgII atom in the [HgCl4]2\u2212 anion is distorted tetra\u00adhedral, with bond angles ranging from 98.16\u2005(3) to 120.68\u2005(3)\u00b0. In the [HgCl4]2\u2212 anion, there are two short Hg\u2014Cl bonds [2.4120\u2005(9) and 2.4171\u2005(11)\u2005\u00c5], one inter\u00admediate Hg\u2014Cl bond [2.4716\u2005(12)\u2005\u00c5] and one long Hg\u2014Cl bond [2.6579\u2005(13)\u2005\u00c5] for the Cl atom involved in a trifurcated hydrogen bond as an acceptor, including two N\u2014H\u22efCl\u22efH\u2014N interactions as well as one C\u2014H\u22efCl inter\u00adaction. There are several C\u2014H\u22efCl inter\u00adactions, with C\u22efCl distances ranging from 3.492\u2005(3) to 3.796\u2005(3)\u2005\u00c5. These link the cations and anions into a zigzag chain along the c-axis direction. In addition, there are Cl\u22efCl halogen bonds, as well as \u03c0\u2013\u03c0 inter\u00adactions, with centroid-to-centroid distances of 3.4765\u2005(18)\u2005\u00c5, which link one of the two benzimidazole moieties into dimeric units.In the title salt, (C H-benzo[d]imidazol-2-yl)benzene have been well explored. This ligand is an ideal candidate for metalation due to the presence of two N atoms and one C atom, which bind tightly with metal atoms . Reports of structurally related complex have been published recently -2-(di\u00adchloro\u00adstiban\u00adyl)-1,3-phenyl\u00adene)bis\u00ad(1-pentyl-1H-benzimidazole) (2) from (4-(tert-but\u00adyl)-2,6-bis\u00ad(1-pentyl-1H-benzimidazol-2-yl)phen\u00adyl)mercury(II) chloride; [C34H41N4HgCl] (1) using SbCl3 in dry 1,4-dioxane via transmetallation. Related reactions to 120.68\u2005(3)\u00b0. In the [HgCl4]2\u2212 anion, there are two short Hg\u2014Cl bonds , one inter\u00admediate Hg\u2014Cl bond and one long Hg\u2014Cl bond for the Cl atom involved in a trifurcated bond as an acceptor including two N\u2014H\u22efCl\u22efH\u2014N interactions as well as one C\u2014H\u22efCl inter\u00adaction , \u2212175.0\u2005(15), 179.7\u2005(15), and \u2212178.1\u2005(9)\u00b0, respectively, while for C30\u2013C34 they are C23\u2014N3\u2014C30\u2014C31, N3\u2014C30\u2014C31\u2014C32, C30\u2014C31\u2014C32\u2014C33, and C31\u2014C32\u2014C33\u2014C34 . Thus the first side chain is in an all-trans conformation while the second side chain has adopted a conformation where it curls up at the end.In the ligand, the dihedral angles between the benzimidazole moieties and central phenyl ring are 40.60\u2005(9) and 38.08\u2005(10)\u00b0, while the angle between them is 36.04\u2005(6)\u00b0. One of the pentyl substituents was refined as disordered over two sets of sites, with occupancies of 0.733\u2005(18)/0.267\u2005(18). The two pentyl side chains have adopted different conformations and this is illustrated by their torsion angles. For C8c-axis direction, as shown in Fig.\u00a03x, \u2212y, 2\u00a0\u2212\u00a0z) with centroid-to-centroid distances of 3.477\u2005(2)\u2005\u00c5.In addition to the inter-ionic hydrogen bonds mentioned above, there are several C\u2014H\u22efCl inter\u00adactions with C\u22efCl distances ranging from 3.492\u2005(3) to 3.796\u2005(3)\u2005\u00c5 (see Table1). These link the cations and anions into a zigzag chain in the n-hexyl rather than n-pentyl side chains for salts containing both the benzimidazole moiety as well as the tetra\u00adchlorido\u00admercurate(II) anion gave eight hits, including a closely related ligand with 1 in dry 1,4-dioxane was added SbCl3 at room temperature. The reaction mixture was refluxed for 6\u2005h under an inert atmosphere of N2 and filtered through Whatman filter paper. When the solvent was evaporated, a white-colored precipitate was obtained and purified by washing with hexane. The compound was dried under vacuum. Colourless block-shaped single crystals were obtained from MeOH at room temperature, yield 64% (0.120\u2005g).The reaction scheme is shown in Fig.\u00a011H NMR : \u03b4 8.13 , 7.91 , 7.82 , 7.49\u20137.45 , 4.43 , 1.74 , 1.43 , 1.14 , 0.72 . 13C NMR : 153.1, 151.4, 137.9, 134.7, 129.4, 128.5, 128.2, 124.7, 124.5, 117.8, 112.6, 45.1, 35.5, 31.3, 29.1, 28.5, 21.9, 14.1.n-pentyl side chains was refined as disordered over two sets of sites, with occupancies of 0.733\u2005(18) and 0.267\u2005(18) and both conformers were constrained to have similar metrical parameters using the SAME command in SHELXL2016. H atoms were positioned geometrically and refined as riding: N\u2014H = 0.88\u2005\u00c5 with Uiso(H) = 1.2Ueq(N); C\u2014H = 0.95\u20130.98\u2005\u00c5 with 1.2Ueq(C) or 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017004303/zl2697sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017004303/zl2697Isup2.hklStructure factors: contains datablock(s) I. DOI: 1538746CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "CD4+NKG2D+ T cells are associated with tumour, infection and autoimmune diseases. Some CD4+NKG2D+ T cells secrete IFN\u2010\u03b3 and TNF\u2010\u03b1 to promote inflammation, but others produce TGF\u2010\u03b2 and FasL to facilitate tumour evasion. Here, murine CD4+NKG2D+ T cells were further classified into NK1.1\u2212CD4+NKG2D+ and NK1.1+CD4+NKG2D+ subpopulations. The frequency of NK1.1\u2212CD4+NKG2D+ cells decreased in inflamed colons, whereas more NK1.1+CD4+NKG2D+ cells infiltrated into colons of mice with DSS\u2010induced colitis. NK1.1\u2212CD4+NKG2D+ cells expressed TGF\u2010\u03b2 and FasL without secreting IFN\u2010\u03b3, IL\u201021 and IL\u201017 and displayed no cytotoxicity. The adoptive transfer of NK1.1\u2212CD4+NKG2D+ cells suppressed DSS\u2010induced colitis largely dependent on TGF\u2010\u03b2. NK1.1\u2212CD4+NKG2D+ cells did not expressed Foxp3, CD223 (LAG\u20103) and GITR. The subpopulation was distinct from NK1.1+CD4+NKG2D+ cells in terms of surface markers and RNA transcription. NK1.1\u2212CD4+NKG2D+ cells also differed from Th2 or Th17 cells because the former did not express GATA\u20103 and ROR\u2010\u03b3t. Thus, NK1.1\u2212CD4+NKG2D+ cells exhibited immune regulatory functions, and this T cell subset could be developed to suppress inflammation in clinics. Ligands of human NKG2D include MHC class I chain\u2010related protein A/B (MICA/MICB) and UL16\u2010binding proteins (ULBPs). Mouse NKG2D ligands include retinoic acid early induced transcript\u20101 , H60 and murine ULBP\u2010like transcript 1 (MULT\u20101) + T cells. NKG2D in association with its adaptor molecules, namely DAP10 or DAP12, transduces signalling by the activation of phosphatidylinositol 3\u2010kinase (PI3K), resulting in cell activation, survival, cytoskeletal rearrangement and release of cytokines and cytotoxic granules NKG2D is an activating receptor expressed on natural killer (NK) cells, CD8+ NKG2D+ T cells are associated with inflammatory diseases, such as rheumatoid arthritis (RA) + NKG2D+ T cells + NK2D+ T cells exhibit Th1\u2010like properties in tissues because of the produced IFN\u2010\u03b3, TNF\u2010\u03b1 and cytolytic granules. IL\u201015 of the microenvironment either in cis or in trans form contributes to the induction of CD4+ NKG2D+ T cell subset CD4+ NKG2D+ T cell population, which is associated in regulatory activities, is normally found in healthy individuals; CD4+ NKG2D+ T cell population is inversely correlated with disease severity in patients with juvenile\u2010onset systemic lupus, suggesting that CD4+ NKG2D+ T cells acts in regulation rather than inflammation + NKG2D+ T cells with regulatory activity is largely dependent on FasL and TGF\u2010\u03b2; hence, this T cell subset features an immunosuppressive property CD4+ NKG2D+ T cell population significantly increased in RAE\u20101\u03b5 transgenic mice, whose RAE\u20101\u03b5 expression was controlled by the CD86 promoter. CD4+ NKG2D+ T cells produced TGF\u2010\u03b2 to down\u2010regulate NKG2D expression on NK cells, whereas Foxp3 was not expressed in the cytoplasm + NKG2D+ T cells are associated with colitis induced by dextran sodium sulphate (DSS) in mice. Furthermore, whether the subsets of CD4+ NKG2D+ T cells with distinct function could be discriminated by additional cell markers remains unclear. Results show that the frequency of NK1.1\u2212 CD4+ NKG2D+ T cells in colon is negatively correlated with colitis induced by DSS, and NK1.1\u2212 CD4+ NKG2D+ T cell differs from NK1.1+ CD4+ NKG2D+ T cells in terms of cell membrane markers and transcriptional RNAs.The number of mouse CD4The following antibodies were obtained from Biolegend or eBioscience : CD3 (17A2), \u03b3\u03b4 (GL3), CD8 (53.67), CD4 (GK1.5), NK1.1 (PK136), NKG2D (CX5), CD107a (1D4B), IFN\u2010\u03b3 (XMG1.2), NKp46 (29A1.4), NKG2A (16A11), Ly49D (4E5), Ly49H (3D10), TGF\u2010\u03b2 (TW7\u201016B4), FasL (MFL3), IL\u201010 (JES5\u201016E3), IL\u201017 (eBio17B7), CD62L (MEL\u201014), CD44 (IM7), granzyme B (NG2B), perforin (eBioOMAK\u2010D), CD25 (PC61.5), Foxp3 (FJK\u201016S), GITR (YGITR 765), CTLA\u20104 (UC10\u20104B9), CD39 (24DMS1), CD69 (LG.3A10), CCR9 (CW\u20101.2), CD28 (E18), T\u2010bet (4B10), GATA\u20103 (16E10A23) and ROR\u2010\u03b3t (AFKJS\u20109), neutralized TGF\u2010\u03b2 antibody (1D11) and RAE\u20101\u03b5 mAb (205001). C57BL/6 and pCD86\u2010RAE\u20101 transgenic mice n\u00a0=\u00a05). All mice were weighed every day. To assess the extent of colitis, loss in bodyweight , stool consistency and blood in the stool were monitored daily by trained individuals blinded to the treatment groups. Disease activity scores are calculated using the total score, which ranged from 0 to 12. The mice were killed on day 8, and the spleens and intestinal tissues were removed for ex\u00a0vivo analysis. All experimental protocols were approved by the Institutional Animal Care and Use Committee of Yangzhou University.Colitis was induced by administration of DSS to drinking water for 7\u00a0days . The tissues were dissected longitudinally, washed completely and cut into smaller pieces. The tissues were then predigested by Hanks\u2019 balanced salt solution (HBSS) with 5\u00a0mM EDTA and 1\u00a0mM DTT at 37\u00b0C for 20\u00a0min. Mixed cell solution was passed through a nylon filter (100\u00a0\u03bcm) and then digested in PBS containing collagenase D (0.5\u00a0g/L), DNase I (0.5\u00a0g/L) and dispase II (3\u00a0g/L) for another 20\u00a0min. The cell suspension was centrifuged, suspended and washed with RPMI 1640 three times. The mixed cells were supplemented with 35% Percoll and then centrifuged to isolate mononuclear cells. Finally, the mononuclear cells were washed with PBS for further study.Cytokine production was determined using an intracellular staining kit (eBioscience). Lymphocytes were cultured with PMA (200\u00a0ng/ml)/ionomycin (2\u00a0\u03bcg/ml) in the presence of brefeldin A (10\u00a0\u03bcg/ml) for 4\u00a0hrs at 37\u00b0C. At the end of the incubation period, the cells were pre\u2010stained with antibody against surface markers. The cells were fixed, permeabilized, stained with cytokine or isotype antibody and analysed by flow cytometry. Lymphocytes were also permeabilized in referenced buffer and incubated with antibodies against Foxp3, T\u2010bet, GATA\u20103 or ROR\u2010\u03b3t.Mouse tissues were embedded in OCT and frozen instantly in liquid nitrogen for cryostat sections. After fixation, the sections were blocked with donkey serum and stained with goat antimouse RAE\u20101\u03b5 antibody . The sections were washed and stained with Alexa\u2010546 labelled donkey anti\u2010goat as secondary antibody. The sections were covered with DAPI . Fluorescence was detected using Eclipse E600 microscope and analysed using NIS\u2010Elements software .\u2212 CD4+ cells from pCD86\u2010RAE\u20101 transgenic mice were negatively isolated using a CD4+ T cell isolation kit. The cells were stained with PE\u2010antimouse NKG2D antibody and magnetic\u2010labelled anti\u2010PE antibody sequentially. The sorted CD4+ NKG2D+ T cells (5\u00a0\u00d7\u00a0105) were injected into the tail vein of DSS\u2010 or PBS\u2010treated mice (n\u00a0=\u00a05) on days 1, 3 and 5. On day 7, the mice were killed to obtain mononuclear colon cells and spleens. Frequencies of CD4+ NKG2D+ T cells were analysed using flow cytometry gated on 7\u2010AAD\u2212 CD45+ NK1.1\u2212 cells.Splenic NK1.1\u2212 CD4+ NKG2D+ and NK1.1+ CD4+ NKG2D+ cells of normal C57BL/6 mice were sorted using flow cytometry. The sorted cells were lysed in TRIzol reagent and reverse\u2010transcribed into complementary DNAs. The DNA profiles were detected by GeneChip\u00ae Mouse Transcriptome Assay 1.0 in Shanghai GIMIX Information Technology Company. Data were analysed by GCBI online software.Splenic NK1.1\u2212 CD45+ CD4+ NKG2D+ cells were analysed. The frequency of CD4+ NKG2D+ cells was significantly down\u2010regulated in the local inflamed colons of DSS\u2010treated mice and CTLA\u20104, but expressed CD39 at low levels profiles were analysed by gene chips. A total of 161 mRNA genes + NKG2D+ cells was determined by costimulation of the MHC II molecule and NKG2D ligand \u2212 CD4+ NKG2D+ cell after coligation of TCR and NKG2D are still being studied.CD4+ NKG2D+ T cells was observed in patients with rheumatoid arthritis + NKG2D+ T cells were negative in grade 1 cervical intraepithelial neoplasia and melanoma patients treated with sorafenib + NKG2D+ T cells of patients with Crohn's disease. Th17 and Th1 represent 30%\u00a0\u00b1\u00a017% and 12%\u00a0\u00b1\u00a05%, respectively, of the lamina propria CD4+ NKG2D+ population + CD4+ NKG2D+ cells produced IFN\u2010\u03b3, IL\u201017 and IL\u201021, indicating that the NK1.1+ CD4+ NKG2D+ subset may resemble human CD161+ CD4+ NKG2D+ cells involved with inflammation The pro\u2010inflammatory role of CD4\u2212 CD4+ NKG2D+ cells showed distinctive markers from NK1.1+ CD4+ NKG2D+ cells. NKp46 \u2212 CD4+ NKG2D+ cells. Given that IL\u201015 stimulations can program T cells to express NK cell markers, such as NKp46 and NKG2A \u2212 CD4+ NKG2D+ cells may not depend on IL\u201015 alone. Induction of the subset in\u00a0vitro requires coligations of TCR and NKG2D. Naturally occurring regulatory CD4+ NKG2D+ cells were also found in humans \u2212 CD4+ NKG2D+ cells could be produced in thymus innately or induced outside.NK1.1\u2212 CD4+ NKG2D+ cells into the colon with colitis decreased, whereas the frequency of NK1.1\u2212 CD4+ NKG2D+ cells in the spleen of mice was enhanced. Under inflammatory conditions, NK1.1\u2212 CD4+ NKG2D+ cells would be redistributed because of the local environment. NK1.1\u2212 CD4+ NKG2D+ T cells expressed low levels of CD39 \u2212 CD4+ NKG2D+ T cells exhibit potential to migrate into colon tissues. The retention of NK1.1\u2212 CD4+ NKG2D+ T cells in spleens partly resulted from the enhanced expression level of NKG2D ligands of splenic macrophages or DCs under the DSS\u2010induced inflammatory situations. Macrophages or DCs would up\u2010regulate NKG2D ligand expression when they are stimulated by Toll\u2010like receptor ligand \u2212 CD4+ NKG2D+ T cells, because pCD86\u2010RAE\u20101 transgenic mice showed higher expression levels of RAE\u20101 in the colons after DSS treatment than those of the wild mice.The infiltration of NK1.1+ NKG2D+ T cells were further categorized into NK1.1\u2212 CD4+ NKG2D+ T and NK1.1+ CD4+ NKG2D+ T subsets. NK1.1\u2212 CD4+ NKG2D+ T cells suppressed colitis induced by DSS in mice via production of TGF\u2010\u03b2. Regulatory NK1.1\u2212 CD4+ NKG2D+ T cells differ from conventional CD4+ CD25+ Foxp3+ cells and do not have features of NK1.1+ CD4+ NKG2D+ T cells in both phenotype and RNA transcription. This article provides biological features of regulatory NK1.1\u2212 CD4+ NKG2D+ T cells of mice and indicated that NK1.1\u2212 CD4+ NKG2D+ T cells may be used for treatment of inflammation\u2010associated diseases.In conclusion, CD4All authors have declared there are no financial conflicts of interest with regard to this work.Figure\u00a0S1 Detection of colonic T and NK cells of mice treated by DSS or PBS.Figure\u00a0S2 Detection of splenic T and NK cells of mice treated by DSS or PBS.Figure\u00a0S3 Detection of splenic CD3+ \u0263\u03b4\u2212 NK1.1+ CD4+ NKG2D+ T of mice treated by DSS or PBS.Figure\u00a0S4 Expression of Foxp3 in splenic NK1.1\u2212CD4+NKG2D+ Foxp3+ cells of CD86\u2010transgenic or wild type mice treated by DSS.Figure\u00a0S5 Production of IFN\u2010\u0263, IL\u201017, and IL\u201021 by NK1.1+ CD4+NKG2D+ cells of mice treated by DSS or PBS.Figure\u00a0S6 Membrane TGF\u2010\u03b2 on NK1.1\u2212CD4+NKG2D+ cells of spleens from mice treated by DSS or PBS.Click here for additional data file."} +{"text": "EXE) treats estrogen receptor positive (ER+) breast cancer in postmenopausal women by inhibiting the estrogen\u2010synthesizing cytochrome P450 CYP19A1. Variability in the severity and incidence of side effects as well as overall drug efficacy may be partially explained by genetic factors, including nonsynonymous variation in CYP19A1, also known as aromatase. The present study identified phase I EXE metabolites in human liver microsomes (HLM) and investigated mechanisms that may alter the extent of systemic estrogen deprivation in EXE\u2010treated women with breast cancer, including whether functional polymorphisms in aromatase cause differential inhibition by EXE and whether EXE metabolites possess anti\u2010aromatase activity. The potency of EXE and ten of its derivatives was measured with HEK293\u2010overexpressed wild type aromatase (CYP19A1*1) using a rapid novel UPLC tandem mass spectrometry method. Of the ten compounds assayed, five were poor inhibitors (IC50\u00a0\u02c3\u00a050\u00a0\u03bcmol/L) of wild type aromatase while five others, including the major metabolite, 17\u03b2\u2010dihydroexemestane (17\u03b2\u2010DHE), exhibited moderate potency, with IC50 values ranging between 1.2 and 7.1\u00a0\u03bcmol/L. The anti\u2010aromatase activity of EXE was also tested with two common allozymes, aromataseThr201Met (CYP19A1*3) and aromataseArg264Cys (CYP19A1*4). Differential inhibition of variant aromatase is unlikely to account for variable clinical outcomes as EXE\u2010mediated inhibition of aromataseThr201Met (IC50\u00a0=\u00a00.86\u00a0\u00b1 0.12\u00a0\u03bcmol/L) and aromataseArg264Cys (IC50\u00a0=\u00a01.7\u00a0\u00b1\u00a00.65\u00a0\u03bcmol/L) did not significantly differ from wild type (IC50\u00a0=\u00a00.92\u00a0\u00b1\u00a00.17\u00a0\u03bcmol/L). Although less potent than the parent drug, these results suggest that active metabolites may contribute to the therapeutic mechanism of EXE.Exemestane ( A key p\u03b2\u2010DHE as a major metabolite in human plasma has been unequivocally confirmed in studies of postmenopausal women taking EXE . Corning and Integrated DNA Technologies manufactured the NADPH regeneration system and oligonucleotide primers, respectively. A QuikChange II Site\u2010Directed Mutagenesis Kit was purchased from Agilent to produce aromatase variant overexpression vectors. The HEK293 cell line was procured from ATCC . G418, penicillin/streptomycin, fetal bovine serum, Opti\u2010MEM, and DMEM supplemented with 4.5\u00a0g/L glucose, 110\u00a0mg/L sodium pyruvate, and L\u2010glutamine was purchased from Invitrogen along with an XCell electrophoresis system. Lipofectamine 2000, PVDF membranes, Pierce BCA protein assay kit, SuperSignal West Femto Maximum Sensitivity Substrate, sodium dodecyl sulfate (SDS), glycine, tris base, ammonium persulfate (APS), goat anti\u2010rabbit HRP\u2010conjugated antibody (cat. No. 31466), and tetramethylethylenediamine (TEMED) were also purchased from Thermo Fisher Scientific. Nonfat dry milk was prepared by BioRad . Sigma\u2010Aldrich supplied Ponceau staining solution, Tween 20, acrylamide/bis\u2010acrylamide solution, 2\u2010mercaptoethanol, estrone, androstenedione substrate, and estrone\u20102,3,4\u201013C3. Rabbit monoclonal anti\u2010aromatase antibody (cat. no. ab124776) was purchased from Abcam .Hangzhou DayangChem Co. supplied the androgens boldenone, testosterone, and 4\u2010andostene\u20103,17\u2010dione for the synthesis of EXE and its analogs. Tokyo Chemical Industry Co. , Thermo Fisher Scientific , and Sigma\u2010Aldrich produced all other reagents (ACS grade or higher) needed for synthesis. Steroid purification required silica columns and thin\u2010layer chromatography plates . LC/MS grade methanol, acetonitrile, and formic acid was purchased from Thermo Fisher Scientific. XenoTech supplied pooled mixed gender human liver microsomes . Previous studies provide detailed descriptions of the synthesis, purification, and NMR\u2010based identity verification of each compound in a chilled Beckman L7\u201065 ultracentrifuge , resuspended in PBS, and stored at \u221280\u00b0C. The relative expression of aromatase was quantitated in triplicate by subjecting 20\u00a0\u03bcg of protein from each overexpressing cell line to SDS\u2010PAGE in a 10% tris\u2010glycine polyacrylamide gel. Following transfer to PVDF for 90\u00a0min at 30\u00a0V, the membrane was blocked overnight at 4\u00b0C in 5% nonfat dry milk, washed for 30\u00a0min in 0.1% Tween, and probed overnight with anti\u2010aromatase primary antibody (1:2500). The next day, the membrane was again washed for 30\u00a0min, and probed with HRP\u2010conjugated goat anti\u2010rabbit antibody (1:7500) for 1\u00a0h at ambient temperature. Following another 30\u00a0min wash, the blot was incubated with SuperSignal West Femto Maximum Sensitivity Substrate per the manufacturer instructions and imaged on a ChemiDoc Imager . Image J software was used to measure band density while Ponceau staining was used to validate even loading between lanes.Stable overexpression of wild type aromatase in HEK293 was driven by a pcDNA3.1/V5\u2010His\u2010TOPO mammalian expression vector as previously described , 5\u00a0\u03bcmol/L androstenedione, a NADPH regeneration system , and 15\u00a0\u03bcg of microsomes from HEK293 overexpressing wild type or variant aromatase were individually incubated with varying concentrations of each steroid. The\u00a0steroid concentrations used for anti\u2010aromatase activity\u00a0assays\u00a0are as follows: EXE (0.05\u201015\u00a0\u03bcmol/L), 17\u03b2\u2010DHE (0.05\u201330 \u03bcmol/L), 17\u03b1\u2010DHE (100\u00a0\u03bcmol/L), 6\u03b1\u2010spirooxiranandrosta\u20101,4\u2010diene\u20103,17\u2010dione (0.05\u201330\u00a0\u03bcmol/L), 6\u03b2\u2010spirooxiranandrosta\u20101,4\u2010diene\u20103,17\u2010dione (0.05\u201340\u00a0\u03bcmol/L), 6\u03b1\u2010methylandrosta\u20101,4,6\u2010triene\u20103,17\u2010dione (0.5\u201330\u00a0\u03bcmol/L), 6\u03b1\u2010methylandrosta\u20101,4\u2010diene\u20103,17\u2010dione (0.2\u201330\u00a0\u03bcmol/L), 6\u2010hydroxymethylandrosta\u20101,4,6\u2010triene\u20103,17\u2010dione (2.5\u2013300\u00a0\u03bcmol/L), 17\u03b2\u2010hydroxy\u20106\u2010hydroxymethylandrosta\u20101,4,6\u2010triene\u20103\u2010one (100\u00a0\u03bcmol/L), 6\u03b1/\u03b2\u2010hydroxy\u20106\u03b1/\u03b2\u2010hydroxymethylandrosta\u20101,4\u2010diene\u20103,17\u2010dione (100\u00a0\u03bcmol/L), and 6\u03b1/\u03b2,17\u03b2\u2010Dihydroxy\u20106\u03b1/\u03b2\u2010hydroxymethylandrosta\u20101,4\u2010diene\u20103\u2010one (100\u00a0\u03bcmol/L). Organic solvent comprised <1% of the total volume of each enzymatic incubation, which proceeded at 37\u00b0C for 2\u00a0h. Reactions were terminated with 50\u00a0\u03bcl of ice cold acetonitrile and centrifuged at 4\u00b0C for 15\u00a0min at 13,200g. Supernatants were collected and spiked with 50\u00a0ng of estrone\u20102,3,4\u201013C3 as an internal standard. An incubation with microsomes derived from non\u2010transfected HEK293 was also performed to serve as a negative control. Aromatization catalyzed by wild\u2010type or variant aromatase was likewise monitored in the presence of vehicle rather than EXE or compounds from the reference library to reflect maximal uninhibited estrone formation. Estrone was measured using a novel 6\u2010min direct detection UPLC/MS/MS method on the Waters Acquity platform using m/z transitions 271.17\u2192133.09 as a marker for estrone and 274.15\u2192162 for estrone\u20102,3,4\u201013C3. Mobile phase (57% methanol in 0.1% formic acid) was infused isocratically from 0 to 4\u00a0min at a flow rate of 0.4\u00a0mL/min. The column was then washed with methanol for 1\u00a0min followed by 1\u00a0min of re\u2010equilibration with mobile phase. Cone and collision voltages were set at 35\u00a0V and 20\u00a0V respectively. Dwell time for both compounds was 0.1\u00a0sec. IC50 values from incubations with wild\u2010type aromatase were calculated for each compound in GraphPad Prism 6 . One\u2010way ANOVA was used to compare the IC50 value for EXE incubated with wild type aromatase with IC50 values for EXE incubated with overexpressed aromatase allozymes.Per 50\u2010\u03bcl incubation containing 50\u00a0\u03bcg of HLM in PBS (pH 7.4), 400\u00a0\u03bcmol/L EXE, and an NADPH regeneration system was placed in a 37\u00b0C water bath for 4\u00a0h before termination with 50\u00a0\u03bcL of cold acetonitrile. After a 15\u2010min refrigerated centrifugation at 13,200g, the supernatant was examined for phase I EXE metabolites. A 10\u2010min UPLC method was used to separate and detect EXE and the ten other reference compounds through multiple reaction monitoring with positive mode electrospray ionization on a Waters ACQUITY UPLC/MS/MS system . The 1.7\u00a0\u03bcm ACQUITY UPLC BEH C18 column used for these analyses was protected by a 0.2\u00a0\u03bcm in\u2010line filter. The UPLC gradient conditions used have previously been described and its major metabolite 17\u03b2\u2010DHE (IC50\u00a0=\u00a04.3\u00a0\u00b1\u00a00.56\u00a0\u03bcmol/L) were potent and moderate inhibitors of aromatase respectively. These results agree with an earlier study which found that 17\u03b2\u2010DHE was approximately 2.6\u2010fold less potent than EXE , exhibiting nearly 5\u2010fold more potency than its 6\u03b2 stereoisomer (IC50\u00a0=\u00a05.7\u00a0\u00b1\u00a01.6\u00a0\u03bcmol/L). 17\u03b1\u2010DHE and three additional compounds exhibited negligible aromatase inhibition with IC50 values exceeding 100\u00a0\u03bcmol/L than EXE in the present study, a large difference in potency also observed by Buzzetti et\u00a0al. (50\u00a0=\u00a03.3\u20137.1\u00a0\u03bcmol/L). In keeping with the observations of Buzzetti et\u00a0al. between wild type enzyme (0.92\u00a0\u00b1\u00a00.17\u00a0\u03bcmol/L), aromataseThr201Met (0.86\u00a0\u00b1 0.12\u00a0\u03bcmol/L), and aromataseArg264Cys (0.97\u00a0\u00b1 0.09\u00a0\u03bcmol/L) in AAA assays normalized for relative aromatase expression (Fig.\u00a0ICion Fig.\u00a0. The preion Fig.\u00a0. One stuion Fig.\u00a0.\u03b2\u2010DHE, 6\u2010HME, 6\u03b1/\u03b2\u2010hydroxy\u20106\u03b1/\u03b2\u2010hydroxy\u2010methylandrosta\u20101,4\u2010diene\u20103,17\u2010dione, and 6\u03b1/\u03b2,17\u03b2\u2010dihydroxy\u20106\u03b1/\u03b2\u2010hydroxymethyl\u2010androsta\u20101,4\u2010diene\u20103\u2010one were identified in incubations of EXE with pooled human liver microsomes through comparison to reference compounds (Fig.\u00a0\u03b2\u2010DHE\u2010glucuronide produced by UGT2B17 (Sun et\u00a0al. \u03b2\u2010DHE is not only the predominant EXE metabolite formed in human liver microsomes, but also capable of inhibiting aromatase with moderate potency suggesting that it may make clinically relevant contributions to the overall response to EXE in women with ER+ breast cancer (Platt et\u00a0al. 17nds Fig.\u00a0 detectedParticipated in research design: Peterson and Lazarus. Conducted experiments: Peterson. Contributed new reagents or analytic tools: Xia, Chen, and Peterson. Performed data analysis: Peterson. Wrote or contributed to the writing of the manuscript: Peterson, Chen, Xia, and Lazarus.None declared."} +{"text": "The structure of this complex was obtained by X-ray diffraction and supported by DFT calculations.The title compound, [Cu(dppaS 24H20NP2S2)(C18H15P)2]\u00b7CH2Cl2 or [Cu(dppaS2)(PPh3)2]\u00b7CH2Cl2, is a neutral mononuclear copper(I) complex bearing an N,N-bis\u00ad(di\u00adphenyl\u00adphospho\u00adrothio\u00adyl)amidate (dppaS2\u2212) ligand and two tri\u00adphenyl\u00adphosphane ligands. The molecular structure shows that the two S atoms of the dppaS2\u2212 ligand [Cu\u2014S = 2.3462\u2005(9) and 2.3484\u2005(9)\u2005\u00c5] and the two P atoms of the two tri\u00adphenyl\u00adphosphane ligands [Cu\u2014P = 2.3167\u2005(9) and 2.2969\u2005(9)\u2005\u00c5] coordinate to the copper(I) atom, resulting in a tetra\u00adhedral coordination geometry. The crystallographically observed mol\u00adecular structure is compared to the results of DFT calculations.The title compound, [Cu(C Several years ago, we reported some emissive copper(I) complexes bearing diphosphane di\u00adsulfide (PPh3)2] bearing the anionic diphos\u00adphane di\u00adsulfide ligand N,N-bis\u00ad(di\u00adphenyl\u00adphospho\u00adrothio\u00adyl)amidate (dppaS2\u2212) and two tri\u00adphenyl\u00adphosphane ligands.Copper(I) complexes have been studied actively because of the abundance of the metal ore and their inter\u00adesting lumin\u00adescent properties (Costa 2)(PPh3)2] shows that the two sulfur atoms of the dppaS2\u2212 ligand and the two phospho\u00adrus atoms of the two tri\u00adphenyl\u00adphosphane ligands coordinate to the copper atom, resulting in a tetra\u00adhedral coordination geometry and 2.3484\u2005(9)\u2005\u00c5, and those between the copper atom and the phospho\u00adrus atoms of the tri\u00adphenyl\u00adphosphane ligands are Cu\u2014P = 2.3167\u2005(9) and 2.2969\u2005(9)\u2005\u00c5. The diphosphine di\u00adsulfide ligand forms a six-membered ring adopting a boat conformation. The bond order of the P6\u2014S2 and P7\u2014S3 bonds are considered to be slightly smaller than two because the lengths of the bonds are considerably longer than general P=S (= 1.91\u2005\u00c5) bond lengths amine (HdppaS2) and potassium tert-butoxide . Tri\u00adphenyl\u00adphosphane and [Cu(CH3CN)4]PF6 were then added to the reaction solution. After the solution had been stirred for one\u2005h at room temperature, a white powder (KPF6) precipitated. The mixture was then filtered. The solution was added to ethanol (20\u2005ml) and the resulting colorless crystals were obtained by filtration. Yield 256\u2005mg (82%). Analysis found: C, 69.43; H, 4.85; N, 1.36%. Calculated for [Cu(dppaS2)(PPh3)2], C60H50NS2P4Cu: C, 69.51; H, 4.86; N, 1.35%. 31P{1H} NMR 34.6 , \u22122.5 . Broadening of the 31P signals of the phospho\u00adrus atoms directly coordinating to the copper atom, which has a large quadrupole moment, has frequently been observed = 1.2Ueq(C) for methyl\u00adene groups, and 0.95\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for aromatic groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017009380/im2478sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989017009380/im2478Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017009380/im2478Isup3.molSupporting information file. DOI: 1557968CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The isomeric title compounds both display three-dimensional supra\u00admolecular architectures arising from N\u2014H\u22efO, C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions. N-[(2-nitro\u00adphen\u00adyl)sulfon\u00adyl]benzamide, (I), and 4-bromo-N-[(4-nitro\u00adphen\u00adyl)sulfon\u00adyl]benzamide, (II), are described (mol\u00adecular formula = C13H9BrN2O5S in each case). The asymmetric unit of (I) contains two independent mol\u00adecules [(IA) and (IB)], while that of (II) contains one mol\u00adecule. The benzoic acid aromatic ring of mol\u00adecule (IA) is disordered due to rotation about the Car\u2014C(=O) bond over two orientations in a 0.525\u2005(9):0.475\u2005(9) ratio. The dihedral angle between the benzene rings is 85.9\u2005(3)\u00b0 in (IA) and 65.22\u2005(19)\u00b0 in (IB), while in (II), the corresponding value is 56.7\u2005(7)\u00b0. In the crystals of (I) and (II), N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions generate three-dimensional networks.The syntheses and crystal structures of the isomeric 4-bromo- N-(aryl\u00adsulfon\u00adyl)aryl\u00adamides have received much attention as they constitute an important class of drugs for treating Alzheimer\u2019s disease -aryl\u00adamides are known to be potent anti-tumour agents against a broad spectrum of human tumour xenografts in mice aryl\u00adamides aryl\u00adamides and (IB), while that of (II)A) and (IB), the ortho-nitro substitution on the benzene\u00adsulfonyl ring is syn to the N\u2014H bond in the central \u2013C\u2014SO2\u2014N\u2014C(O)\u2013 segment is disordered due to rotation about the Car\u2014C(=O) bond over two orientations in a 0.525\u2005(9):0.475\u2005(9) ratio, which are inclined to each other by 45.5\u2005(4)\u00b0. The nitro groups in both the A and B mol\u00adecules of (I)A) is 56.3\u2005(4)\u00b0, while in (IB), the torsion angle C14\u2014C15\u2014N4\u2014O9 is 35.6\u2005(5)\u00b0, whereas in (II)A), 65.22\u2005(19)\u00b0 in (IB) and 56.7\u2005(7)\u00b0 in (II)S(7) motif.The asymmetric unit of (I)nt Fig.\u00a01. The benN1\u22efO6 and N3\u2014HN3\u22efO4 \u2005\u00c5]. Further, C10\u2014H10B\u22ef\u03c01 [where \u03c01 is the nitro\u00adbenzene ring of mol\u00adecule (IB)] and C12\u2014H12A\u22ef\u03c02 [\u03c02 is the bromo\u00adbenzene ring of mol\u00adecule (IA)] extend the zigzag sheets into a three-dimensional architecture, which is consolidated by several aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions .The crystal structure of (I)4 Table\u00a01 between es Fig.\u00a03. The A +s Table\u00a01. A dimers Table\u00a01 and the N1\u22efO3 hydrogen bonds forming C(4) chains along [100] (Table\u00a02C(5) chains. The mol\u00adecules of neighbouring chains are inter\u00adlinked via C3\u2014H3\u22efO4 and C12\u2014H12\u22efO4 inter\u00adactions (i.e. O4 acts as a double acceptor) and thus, a zigzag sheet propagates in the ac plane (Table\u00a02C(13) and C(5) chains, respectively, along [001]. Mol\u00adecules in adjacent layers are linked via C9\u2014H9\u22efO2 and C10\u2014H10\u22efO1 inter\u00adactions that form C(7) and C(8) chains propagating along the b-axis direction, and thus a three-dimensional network is obtained. A short O5\u22efBr1 [3.173\u2005(4)\u2005\u00c5] contact is observed.The crystal structure of (II)] Table\u00a02: these c] Table\u00a02 forming e Table\u00a02. The C12et al., 2016N-benzoyl\u00adbenzene\u00adsulfonamide Uiso = 1.2Ueq(parent atom).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017001578/hb7646sup1.cifCrystal structure: contains datablock(s) I, II, shelx. DOI: 10.1107/S2056989017001578/hb7646Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017001578/hb7646IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989017001578/hb7646Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989017001578/hb7646IIsup5.cmlSupporting information file. DOI: 1530208, 1530207CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of tetra\u00adiso\u00adbutyl\u00adthiuram di\u00adsulfide reveals a \u221285.81\u2005(1)\u00b0 C\u2014S\u2014C\u2014S torsion angle and multiple intra- and inter\u00admolecular S\u22efC\u2014H close contacts. 18H36N2S4, crystallizes in a general position in the triclinic space group P-1 but shows pseudo-C2 symmetry about the di\u00adsulfide bond. The C\u2014S\u2014S\u2014C torsion angle [\u221285.81\u2005(2)\u00b0] and the dihedral angle between the two NCS2 mean planes [85.91\u2005(5)\u00b0] are within the range observed for this compound type. Multiple intra- and inter\u00admolecular S\u22efH\u2014C close contacts appear to play a role in assisting the specific conformation of the pendant isobutyl groups and the packing arrangement of mol\u00adecules within the cell. Tetra\u00adiso\u00adbutyl\u00adthiuram di\u00adsulfide mol\u00adecules of one optical configuration form sheets in the plane of the a and b axes. Inversion centers exist between adjoining sheets, which stack along the c axis and alternate in the handedness of their constituent mol\u00adecules.Tetra\u00adkis(2-methyl\u00adprop\u00adyl)thio\u00adper\u00adoxy\u00addicarbonic di\u00adamide, or tetra\u00adiso\u00adbutyl\u00adthiuram di\u00adsulfide, C N,N,N\u2032,N\u2032-Tetra\u00adalkyl\u00adthio\u00adper\u00adoxy\u00addicarbonic di\u00adamides, com\u00admon\u00adly called tetra\u00adthiuram di\u00adsulfides, comprise a class of organosulfur compounds with applications that are both diverse and long-standing. Tetra\u00admethyl\u00adthiuram di\u00adsulfide, known by the commercial name thiram, is broadly useful both as a fungicide 2] that was more complex than anti\u00adcipated, even considering the hindered rotation about the \u2212S2\u2013CNiBu2 bond thio\u00adper\u00adoxy\u00addicarbonic di\u00adamide (tetra\u00adiso\u00adbutyl\u00adthiuram di\u00adsulfide) itself does not show evidence of such intra\u00admolecular inter\u00adaction, several recent studies of tetra\u00adthiuram di\u00adsulfides have suggested such inter\u00adactions in the crystalline state . Despite the lack of strict C2 symmetry, tetra\u00adiso\u00adbutyl\u00adthiuram di\u00adsulfide is nevertheless chiral. The image in Fig.\u00a01a presents the mol\u00adecule with a left-handed configuration to the core \u2013H2CNC(S)S\u2013SC(S)NCH2\u2013 portion. If Fig.\u00a01a were to be viewed from above, along the pseudo C2 axis that bis\u00adects the S3\u2014S4 bond, the C1\u2014S1 and C2\u2014S2 thione bonds would project forward and backward, respectively, from the plane of the paper and thereby define a left-handed propeller. The right-handed counterpart is necessarily the other occupant of the unit cell, as required by the racemic space group. Among the structurally characterized thiuram di\u00adsulfides, crystallographically imposed C2 symmetry is also common \u2005\u00c5, while the thione C=S bonds are essentially identical at 1.642\u2005(3) and 1.643\u2005(3)\u2005\u00c5. The C1\u2014S3\u2014S4\u2014C2 torsion angle, \u03c4, is \u221285.81\u2005(2)\u00b0 and, as is typical of tetra\u00adthiuram di\u00adsulfides, very similar in magnitude to the angle of 85.91\u2005(5)\u00b0 between the mean planes defined by the Sb and shown in Table\u00a01D\u2014H\u22efA angles are closer to 90\u00b0 than to 180\u00b0 , 3.810\u2005(3)\u2005\u00c5]. The geometric parameters for both these intra\u00admolecular and inter\u00admolecular S\u22efC\u2013H contacts fall within the range defined as consistent with a weak D\u2014H\u22efA inter\u00adaction and the dihedral angle (\u03b8) between Set al., 20092Cl2 solution. 1H NMR : 3.83 , 2.39 , 0.98 , 0.87 .The synthesis procedure employed was that described by Kapanda 2) or 1.5 times (for \u2013CH3) those of the carbon atoms to which they were attached. The C\u2014H distances assumed were 1.00, 0.99, and 0.98\u2005\u00c5 for the \u2013CH\u2013, \u2013CH2, and \u2013CH3 types of hydrogen atoms, respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017015158/lh5851sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017015158/lh5851Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017015158/lh5851Isup3.cmlSupporting information file. DOI: 1580550CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Solvents are the source of the protons at the ylidic C atom.Reaction of BI 3 with carbodi\u00adphospho\u00adrane, C(PPh3)2, gives a mixture of the dicationic compounds, methyl\u00adenebis(tri\u00adphenyl\u00adphospho\u00adnium) diiodide di\u00adchloro\u00admethane disolvate, C37H32P22+\u00b72I\u2212\u00b72CH2Cl2 or [Ph3PCH2PPh3]I2\u00b72CH2Cl2 (I), methyl\u00adenebis(tri\u00adphenyl\u00adphospho\u00adnium) bis\u00ad(tetra\u00adiodo\u00adborate), C37H32P22+\u00b72BI4\u2212 or [Ph3PCH2PPh3](BI4)2 (II). Solvents are the source of the protons at the ylidic C atom. The P\u2014C\u2014P angle is 124.1\u2005(2)\u00b0 for (I) and 121.7\u2005(3)\u00b0 for (II), while the two P\u2014C bond lengths are 1.804\u2005(4) and 1.807\u2005(5)\u2005\u00c5 in (I), and 1.817\u2005(5) and 1.829\u2005(5)\u2005\u00c5 in (II). In the crystal of (I), the protons of the central P\u2014CH2\u2014P C atom exhibit weak C\u2014H\u22efI hydrogen bonds with the respective anions. The anions in turn are linked to the di\u00adchloro\u00admethane solvent mol\u00adecules by C\u2014H\u22efI hydrogen bonds. In the crystal of (II), one of the BI4\u2212 anions is linked to a phenyl H atom via a weak C\u2014H\u22efI hydrogen bond.Reaction of BI The volume of the resulting solution was reduced to ca 3\u2005ml and layered with ca 5\u2005ml of hexane. A crop of crystals formed in a few days .(PhUiso(H) = 1.2Ueq(C). For both compounds, a small number of reflections were affected by the beam stop and were omitted from the final cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989017010295/su5375sup1.cifCrystal structure: contains datablock(s) Global, II, I. DOI: 10.1107/S2056989017010295/su5375Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017010295/su5375IIsup4.hklStructure factors: contains datablock(s) II. DOI: 1533031, 1552112CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The compound comprises an octa\u00adhedral homoleptic Me2SO-solvated cobalt(II) cation and a tetra\u00adhedral cobaltate(II) anion attached to three chloro ligands and one quinoline moiety.Anhydrous cobalt(II) chloride reacts with quinoline (C 2SO) or quinoline (C9H7N). The title compound, [Co(C2H6OS)6][CoCl3(C9H7N)]2, is a cobalt salt in which the metal ion is complexed to both Me2SO and quinoline. In particular, we observed that anhydrous cobalt(II) chloride reacts with quinoline in Me2SO to form a salt that is to be formulated as [CoII(Me2SO)6]2+{[CoIICl3quinoline]2\u2212}. The CoII atom in the cation portion of this mol\u00adecule lies on a inversion center and is bound to the O atoms of six Me2SO moieties in an octa\u00adhedral configuration, while the CoII atom in the anion is attached to three chloride ligands and one quinoline moiety in a tetra\u00adhedral arrangement.There are few reports that describe crystal structures of compounds containing cobalt complexed to either dimethyl sulfoxide (Me A closely related complex, [Co(Me2SO)6][CoCl4], contains a cobalt cation that is similarly surrounded by six oxygen atoms in a slightly distorted octa\u00adhedral configuration with Co\u2014O distances between 2.06\u2005(1) and 2.10\u2005(1)\u2005\u00c5, with a mean Co\u2014O distance of 2.08\u00c5 bond angles in the title complex are close to 90\u00b0, ranging from 86.29\u2005(7) to 93.71\u2005(7)\u00b0, compared to 87.9\u2005(5) to 90.8\u2005(4)\u00b0 in [Co(Me2SO)6][CoCl4] 6][CoCl4] also showed some distortion with Co\u2014Cl distances ranging from 2.265\u2005(6) to 2.305\u2005(7)\u2005\u00c5, giving an average Co\u2014Cl distance of 2.284\u2005(6)\u2005\u00c5, and the Cl\u2014Co\u2014Cl angles ranging from 107.1\u2005(2) to 112.4\u2005(2)\u00b0 6][CoCl4] were ascribed to disorder, as indicated by the high anisotropic motion to 2.2534\u2005(10)\u2005\u00c5, with an average Co\u2014Cl distance of 2.252\u2005\u00c5, while the Co\u2014N distance is 2.054\u2005(3)\u2005\u00c5. The Cl\u2014Co\u2014Cl angles range from 108.21\u2005(5) to 114.26\u2005(4)\u00b0, giving an average of 110.98\u00b0, and the average N\u2014Co\u2014Cl angle is 107.88\u00b0 [range 107.09\u2005(9) to 108.80\u2005(8)\u00b0], indicating that while the anion is close to tetra\u00adhedral, there is some distortion. Inter\u00adestingly, the [CoCl4 index that is reported and discussed elsewhere ]/141, where \u03b1 and \u03b2 represent the two largest angles; a \u03c44 value of 1.00 indicates an idealized tetra\u00adhedral geometry, whereas a value of 0.00 indicates an idealized square-planar geometry. In the title complex, \u03b1 = 114.26\u2005(4)\u00b0 and \u03b2 = 110.46\u2005(4)\u00b0, such that \u03c44 is 0.96, which indicates very little deviation from a tetra\u00adhedral geometry. For comparison, \u03c44 for the [CoCl4]2\u2212 anion in [Co(Me2SO)6][CoCl4] is 0.98 6]2+ and [CoIICl3quinoline]\u2212 ions, with the exception of very weak C\u2014H\u22efCl interactions. The distances between the Cl and the carbon atoms of the methyl groups of the Me2SO ligands are, for example, Cl1\u22efC32\u2014S3 [3.525\u2005(3)\u2005\u00c5], Cl1\u22efC31\u2014S3 [3.736\u2005(4)\u2005\u00c5], Cl2\u22efC22\u2014S2 [3.633\u2005(4)\u2005\u00c5], Cl2\u22efC21\u2014S2 [3.770\u2005(4)\u2005\u00c5], Cl3\u22efC12\u2014S1 [3.638\u2005(4)\u2005\u00c5] and Cl3\u22efC32\u2014S3 [3.819\u2005(4)\u2005\u00c5] and are comparable to the sum of the van der Waals radii of Cl and CH3 of 3.80\u2005\u00c5 6][CoCl4] complex as discussed above 6][CoCl4] 6][CoCl4], there are a few other examples of cobalt complexes solvated by Me2SO that are listed in the Cambridge Database 6][ClO4]2, one of which possesses Co\u2014O distances in the range 2.0833\u2005(17)\u20132.0934\u2005(15)\u2005\u00c5, giving a mean Co\u2014O distance of 2.088\u2005(5)\u2005\u00c5, with O\u2014Co\u2014O (cis) angles between 90.11\u2005(6) and 92.31\u2005(6)\u00b0 angles between 85.26\u2005(7) and 93.67\u2005(8)\u00b0 6][SnCl6], both the cobalt and tin metal ions display an octa\u00adhedral environments, with the Co\u2014O bond lengths reported between 2.093\u2005(4) and 2.113\u2005(5)\u2005\u00c5 angles vary between 89.0\u2005(2) and 90.0\u2005(2)\u00b0 6][NO3]3 is also known and possesses six equivalent Co\u2014O bond lengths of 2.005\u2005(2)\u2005\u00c5, which are shorter than the values in the CoII complexes (Me2SO-S)(C2H5OH)]\u00b7C2H5OH (acv = a\u00adcyclo\u00advir) is rare, there are some notable examples. For example, the compound bis\u00ad(dimethyl sulfoxide)\u00adhydridobis(tri\u00adphenyl\u00adphosphane)cobalt(I), [CoH(C18H15P)2(Me2SO)2], contains CoI coordinating a hydride anion, two phosphine ligands and two Me2SO moieties that are bound through the sulfur atom in a distorted trigonal\u2013bipyramidal structure -porphyrinato)-cobalt(III) bis\u00ad(dimethyl sulfoxide-\u03baS)-porph\u00adyr\u00adinato)cobalt(III) bis\u00ad(hexa\u00adfluoro\u00adanti\u00admonate) dimethyl sulfoxide solvate 2]\u2212 of the title compound is 2.037\u2005(5)\u2005\u00c5 while the Co\u2013Cl bond lengths are 2.2517\u2005(10)\u20132.2534\u2005(10)\u2005\u00c5, and the Cl\u2014Co\u2014Cl and Cl\u2014Co\u2014N angles range between 108.21\u2005(5) and 114.26\u2005(4)\u00b0, and 107.09\u2005(9) and 108.80\u2005(8)\u00b0, respectively. For comparison, the Co\u2014N bond lengths in the CoIICl2(quinoline)2 complex are 2.061\u2005(3) and 2.037\u2005(5)\u2005\u00c5 and the Co\u2014Cl bond lengths are 2.246\u2005(2) and 2.241\u2005(1)\u2005\u00c5 in Me2SO (20\u2005mL) and refluxed for one\u2005h. After cooling down, the mixture was transferred to a beaker and placed in a desiccator containing anhydrous calcium chloride pellets (4\u201320 mesh) to crystallize over a period of four months. Deep-blue crystals of [Co(Me2SO)6]2+{[CoCl3quinoline]2}\u2212 suitable for X-ray diffraction were obtained from this process of slow evaporation. Notably, when the reaction between anhydrous cobalt(II) chloride and quinoline is conducted in EtOH, rather than Me2SO, the previously reported [CoIICl2(quinoline)2] complex is obtained chloride was mixed with quinoline, CUiso(H) = 1.2Ueq(Csp2) or 1.5Ueq(Csp3).Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2056989018001652/lh5868sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018001652/lh5868Isup2.hklStructure factors: contains datablock(s) I. DOI: 1820336CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The synthesis and structural determination of bis\u00adsulfone tetra\u00ad(nitrate) monohydrate is reported. The crystal structure features N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen-bonds and \u03c0\u2013\u03c0 inter\u00adactions. 12H14N2O2S2+\u00b74NO3\u2212\u00b7H2O {alternative name: bis[bis\u00ad sulfone] tetra\u00adnitrate monohydrate}, the cations are conformationally similar, with comparable dihedral angles between the two benzene rings in each of 70.03\u2005(18) and 69.69\u2005(19)\u00b0. In the crystal, mixed cation\u2013anion\u2013water mol\u00adecule layers lying parallel to the (001) plane are formed through N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen-bonding inter\u00adactions and these layers are further extended into an overall three-dimensional supra\u00admolecular network structure. Inter-ring \u03c0\u2013\u03c0 inter\u00adactions are also present [minimum ring centroid separation = 3.693\u2005(3)\u2005\u00c5].In the title compound, the hydrated tetra\u00ad(nitrate) salt of dapsone , 2C Ka ca 2), is a drug that has been used to treat a diversity of diseases including tuberculosis, leprosy, malaria and AIDS-related pneumonia , a very weak Lewis base anilinium 2-carb\u00adoxy-4,6-di\u00adnitro\u00adphen\u00adolate monohydrate has been reported sulfone cations (A and B), four nitrate anions and one water mol\u00adecule (O1W) in the asymmetric unit (Aa) and (N1/C7\u2013C12) (Ab)] and cation B [defined by (N3/C13\u2013C18) (Ba) and (N4/ C19\u2013C24) (Bb)] are 70.03\u2005(18) and 69.69\u2005(19)\u00b0, respectively. As expected the anilinium groups are planar with maximum r.m.s. deviations of 0.0044, 0.0120, 0.0114 and 0.0072\u2005\u00c5, respectively.The title compound crystallizes in the ortho\u00adrhom\u00adbic space group it Fig.\u00a01. The di\u00advi and nitro O12vii acceptors. The two cations A and B are associated through \u03c0\u2013\u03c0 inter\u00adactions [ring centroid separation CgAb\u22efCgBai = 3.693\u2005(3)\u2005\u00c5 [symmetry code: (i) \u2212x\u00a0+\u00a01, y\u00a0+\u00a0z\u00a0+\u00a0a-axis direction bond motif motifs 3\u00b79H2O in EtOH (2\u2005ml) was added dropwise to 4,4\u2032-di\u00adamino\u00addiphenyl sulfone in EtOH (5\u2005ml), with continuous stirring at room temperature for 72\u2005h. Slow evaporation of this solution yielded yellow crystals suitable for X-ray analysis within 5\u2005d.Fe(NOUiso(H) = 1.2Ueq(C). All N\u2014H atoms were located by difference methods but were subsequently restrained in the refinement with N\u2014H = 0.89\u2005\u00c5 and Uiso = 1.2Ueq(N). The H atoms of the water mol\u00adecule were also located in a Fourier map and were allowed to ride with a restrained O\u2014H bond length = 0.85\u2005(1)\u2005\u00c5 and H\u22efH = 1.39\u2005(2)\u2005\u00c5 and Uiso(H) = 1.5 Ueq(O). Although not of relevance in this achiral compound, the Flack absolute structure parameter I. DOI: 10.1107/S2056989017014803/zs2390Isup2.hklStructure factors: contains datablock(s) I. DOI: 1579678CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Their structures and absolute configurations were unambiguously established by a combination of NMR and MS analysis and electronic circular dichroism (ECD)\u00a0evidence. Callisalignenes H (2) and I (3) have a rare sec-butyl moiety at C-7. Meroterpenoids 1\u20133 exhibited cytotoxicity against HCT116 cells with IC50 values of 8.51\u00a0\u00b1\u00a01.8, 9.12\u00a0\u00b1\u00a00.3, and 16.33\u00a0\u00b1\u00a03.3\u00a0\u03bcM, respectively.Callisalignenes G\u2013I contains supplementary material, which is available to authorized users. Callistemon (Myrtaceae) are evergreen shrubs or small trees native to Australia and have been popularly cultivated in southern China as ornamental species with bottle brush inflorescence. Recently, acylphloroglucinols and their derivatives, including adducts of a phloroglucinol moiety coupled with its derivative or a terpenoid unit, have been extensively\u00a0obtained from this genus . Further analysis of its experimental and calculated ECD spectra -4.Besides the three new meroterpenoids, two known compounds were identified as (\u2013)-callistenone F (4) and vimiS. aureus, E. coli, and P. aeruginosa) and three fungal strains . None of them showed antimicrobial effects (MIC\u00a0>250\u00a0\u03bcg/mL). Additionally, cytotoxicities of 1\u20135 against six human cancer cells were also conducted\u00a0and the results were summarized in Table 1\u20133 exhibited cytotoxicity against HCT116 cells with IC50 values of 8.51\u00a0\u00b1\u00a01.8, 9.12\u00a0\u00b1\u00a00.3, and 16.33\u00a0\u00b1\u00a03.3\u00a0\u03bcM, respectively, compared to that of positive control . Moreover, 1 and 3 displayed cytotoxicity against A549 cells with IC50 values of 12.85\u00a0\u00b1\u00a08.2 and 10.03\u00a0\u00b1\u00a03.2\u00a0\u03bcM , respectively.All the isolates were evaluated for their antimicrobial effects toward three bacterial (cts MIC\u00a0>50\u00a0\u03bcg/mL.\u03b4) were expressed in ppm with reference to the solvent signals, and coupling constant (J) values were reported in Hz. HRESIMS data were measured using an Agilent 1290 UPLC/6540 Q-TOF mass spectrometer. Sephadex LH-20 , Si gel , and RP-18 were used for column chromatography (CC). Semi-preparative HPLC was performed on an Agilent 1260 instrument with a ZORBAX SB-C18 column . Fractions were monitored by Si gel GF254 or RP-18 F254 plates. Spots were visualized under UV light and by spraying with 10% H2SO4 in EtOH followed by heating.Optical rotations were measured on a Jasco P-1020 polarimeter. UV spectra were recorded on a Shimadzu UV2401 PC spectrophotometer. IR spectra were determined on Bruker FT-IR Tensor-27 infrared spectrophotometer with KBr discs. ECD spectra were recorded on an Applied Photophysics spectropolarimeter. 1D and 2D NMR spectra were recorded on Bruker AV 600 or 800\u00a0MHz spectrometers using TMS as an internal standard. Chemical shifts . A voucher specimen (HY0025) was deposited in the State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences.Twigs and leaves of C. salignus (10.0\u00a0kg) were percolated with petroleum ether (PE) at room temperature three times and then filtered. After removal of solvent under reduced pressure, the crude extract (130\u00a0g) was subjected to silica gel CC, eluted with PE-EtOAc to yield five fractions A\u2013E. Fraction A (25\u00a0g) was applied to Sephadex LH-20 column to give A3 (3\u00a0g), which was further separated on an RP-18 column and eluted with a gradient of MeCN\u2013H2O to obtain five subfractions (A3-1\u2013A3-5). Subfraction A3-4 (225\u00a0mg) was subsequently purified by semi-preparative HPLC with MeCN\u2013H2O as mobile phase to afford 1 (5\u00a0mg), 2 (4\u00a0mg), 3 (8\u00a0mg), and 5 (35\u00a0mg). Similarly, fraction C (42\u00a0g) was separated by Sephadex LH-20 column to give three subfraction (C1\u2013C3). After repeated purification by RP-18 column with MeCN\u2013H2O , 4 (18\u00a0mg) was obtained from C2 (0.5\u00a0g).Air-dried and powdered twigs and leaves of \u03b1] +188.3 ; UV (MeOH) \u03bbmax (log \u03b5) 265 (4.18) nm; CD (MeOH) 265 (\u0394 \u03b5 + 36.23), 306 (\u0394\u03b5 \u22127.57) nm; IR (KBr) vmax 3440, 2932, 1721, 1651, 1469, 1248\u00a0cm\u20131; 1H and 13C NMR data, see Table\u00a0m/z 409.2714 [M\u00a0+\u00a0Na]+ .Colorless gum; [\u03b1] \u2212221.4 ; UV (MeOH) \u03bbmax (log \u03b5) 266 (4.20) nm; CD (MeOH) 267 (\u0394\u03b5 \u221231.35), 309 (\u0394\u03b5 + 4.11) nm; IR (KBr) vmax 3439, 2930, 1720, 1649, 1467, 1247\u00a0cm\u22121; 1H and 13C NMR data, see Table\u00a0m/z 409.2708 [M\u00a0+\u00a0Na]+ .Colorless gum; [\u03b1] +164.7 ; UV (MeOH) \u03bbmax (log \u03b5) 267 (4.24) nm; CD (MeOH) 267 (\u0394\u03b5 +27.50), 306 (\u0394\u03b5 \u22126.02) nm; IR (KBr) vmax 3428, 2962, 1718, 1651, 1467, 1384, 1174\u22121; 1H and 13C NMR data, see Table\u00a0m/z 387.2899 [M\u00a0+\u00a0H]+ .Colorless gum; [\u03b1] \u2212133.0 ; CD (MeOH) 235 (\u0394\u03b5 +5.45), 248 (\u0394\u03b5 +1.77), 286 (\u0394\u03b5 +15.10), 317 (\u0394\u03b5 \u221213.95) nm.Colorless gum; [The conformations generated by the MM2 force field in Chem-Bio3D software overlaid with key correlations observed in the ROESY spectrum were subjected to semi-empirical PM3 quantum mechanical geometry optimizations using the Gaussian 09 program . The cor1\u20135 were carried out against three bacterial strains and three fungal strains using the antimicrobial susceptibility assay [The antimicrobial activities compounds ty assay . The comAll the compounds were evaluated for their cytotoxicities against six tumor cell lines, including HCT116 , Huh7 (human hepatoma cell line), Hela , CCRF-CEM (human acute lymphocytic leukemia cell line), DU145 (human prostatic cancer cell line), and A549 (human lung cancer cell line), by MTT assay in 96-well plates [Supplementary material 1 (DOCX 12178\u00a0kb)Below is the link to the electronic supplementary material."} +{"text": "L3Phen (where L\u2212 is the sulfonyl\u00adamido\u00adphosphate (SAPh)-type ligand N-{bis\u00ad[meth\u00adyl(phen\u00adyl)amino]\u00adphosphor\u00adyl}benzene\u00adsulfonamidate, C6H5SO2NHPO[N(CH3)C6H5]2 has been synthesized and its crystal structure determined.A lanthanum(III) complex with formula La 20H21N3O3PS)3(C12H8N2)] is created by one LaIII ion, three deprotonated N-{bis\u00ad[meth\u00adyl(phen\u00adyl)amino]\u00adphosphor\u00adyl}benzene\u00adsulfonamidate (L\u2212) ligands and one 1,10-phenanthroline (Phen) mol\u00adecule. Each LaIII ion is eight-coordinated (6O+2N) by three phosphoryl O atoms, three sulfonyl O atoms of three L\u2212 ligands and two N atoms of the chelating Phen ligand, leading to the formation of six- and five-membered metallacycles, respectively. The lanthanum coordination polyhedron has a bicapped trigonal\u2013prismatic geometry. \u2018Sandwich-like\u2019 intra\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions are observed between the 1,10-phenanthroline ligand and two benzene rings of two different L\u2212 ligands. The phenyl rings of L\u2212 that are not involved in the stacking inter\u00adactions show minor positional disorder. Mol\u00adecules form layers parallel to the (010) plane due to weak C\u2014H\u22efO inter\u00admolecular hydrogen bonds. Unidentified highly disordered solvate mol\u00adecules that occupy ca 400\u2005\u00c53 large voids have been omitted from the refinement model.The asymmetric unit of [La(C These types of compounds were first synthesized by Kirsanov , containing the functional fragment C(O)NHP(O), because of their properties as extractants (OMe)2] amido\u00adphosphate [(Me)PhSO2NHP(O)(OMe)2] and dimethyl 2-naphthyl\u00adsulfonyl\u00adamido\u00adphosphate [(C10H7)SO2NHP(O)(OMe)2] indicate that the ligand first excited singlet state plays a dominant role in intra\u00admolecular energy transfer processes in these Ln complexes amido\u00adphosphate [PhSON-(meth\u00adyl(phenyl\u00adamino)\u00adphosphor\u00adyl)benzene\u00adsulfon\u00adamide (HL) [PhSO2NHP(O)(N(Me)Ph)2] with the general formula 3PhenLa(L).Knowledge of the crystal structure is an essential part of understanding the luminescent properties of these types of lanthanide complexes. In this paper we would therefore like to report the mol\u00adecular and crystal structure of a lanthanum coordination compound based on the amidic type SAPh ligand 3PhenLa(L) crystallizes with one mol\u00adecule in the asymmetric unit bond lengths [2.516\u2005(2)\u20132.541\u2005(2)\u2005\u00c5] are all longer than those of their La\u2014O(P) counterparts [2.424\u2005(2)\u20132.463\u2005(2)\u2005\u00c5], with mean values of 2.435 and 2.456\u2005\u00c5, respectively. The mean average of all La\u2014O bond lengths is 2.476\u2005\u00c5. The La\u2014N distances are with 2.699\u2005(3) and 2.700\u2005(3)\u2005\u00c5 shorter than those previously obtained for a 1,10-phenanthrolinate lanthanum (III) complex with hexa\u00adfluoro\u00adacetyl\u00adacetonate \u20131.474\u2005(2)\u2005\u00c5 and 1.491\u2005(2)\u20131.494\u2005(2)\u2005\u00c5 in their expected ranges. The mean values are 1.468 and 1.492\u2005\u00c5, respectively. The corresponding bond lengths in the related neutral ligands are around 1.42\u2005\u00c5 (Moroz A) and La1\u2013O3\u2013P3\u2013N3\u2013S3\u2013O9 (B) rings both adopt twist-boat conformations are 0.78 and 0.41\u2005\u00c5, respectively. The deviations of the La1 and O3 atoms from the mean plane through the remaining atoms of B (r.m.s.deviation = 0.06\u2005\u00c5) are 0.9 and 0.88\u2005\u00c5, respectively. The La1\u2013O2\u2013P2\u2013N2\u2013S2\u2013O7 (C) ring adopts a flattened half-chair conformation . The deviation of the La1 atom from the mean plane carried through the remaining atoms of ring C (r.m.s.deviation 0.02\u2005\u00c5) is 0.36 \u2005\u00c5.The six-membered metallocyclic rings with the chelate (O)PNS(O) fragments are all non-planar. The La1\u2013O1\u2013P1\u2013N1\u2013S1\u2013O1 similar to that of the Tb(Pip)3(Phen) mixed-ligand complex with 2,2,2-tri\u00adchloro-N-(dipiperidin-1-yl-phosphor\u00adyl)acetamide, HPip \u22efC69(\u03c0) of the 1,10-phenanthroline mol\u00adecule inter\u00adacts with the C41(\u03c0)\u22efC46(\u03c0) phenyl ring at the sulfonyl group from another ligand , and with the C8(\u03c0)\u22efC13(\u03c0) ring at the phosphoryl group from the other ligand . A similar intra\u00admolecular organization was described previously for related compounds (Beloso 3PhenLa(L) mol\u00adecules are linked by weak C\u2014H\u22efO hydrogen bonds .In the crystal phase, the s Table\u00a01, formings Table\u00a01. There aet al., 2016N-(bis\u00ad(di\u00adamino)\u00adphosphor\u00adyl)sulfonamide fragments yielded five hits, with only one metal complex structure with a neodymium metal atom among them --erbium(III) phospho\u00adramidato-2O6 environment yielded 20 hits, with average La\u2014O and La\u2014N bond lengths of 2.476 and 2.693\u2005\u00c5, respectively. 11 complex structures with different lanthanoid metals (Ln) containing Ln\u2013O\u2013P\u2013N\u2013S\u2013O metallocycles were found in the database, all with octa\u00adcoordinated metal atoms. Most of those metallacyclic rings are non-planar with mean deviations of the O and N atoms of 0.329 and 0.434\u2005\u00c5, respectively.A search for octa\u00adcoordinated La complexes with an LaN1H and 31P NMR spectra in DMSO-d6 solutions were recorded on a Varian 400 NMR spectrometer at room temperature. 1H chemical shifts were determined relative to the inter\u00adnal standard TMS whereas 31P chemical shifts were determined relative to 85% H3PO4 as an external standard. Infrared (FTIR) spectra were recorded on a Perkin\u2013Elmer Spectrum BX spectrometer using KBr pellets. The resolution of the FTIR spectra is 1\u2005cm\u22121.Sulfonyl\u00adamido\u00adphosphate ligandN-(meth\u00adyl(phenyl\u00adamino)\u00adphosphor\u00adyl)benzene\u00adsulfonamide (HL) was synthesized via a three-step procedure based on the Kirsanov reaction \u03b4 2.95 , 7.05 , 7.14 , 7.21 , 7.56 , 7.65 , 7.91 .\u22121): 3062 , 2948 , 2780 , 2705 , 2655 , 1594 , 1495 (s), 1446 (m), 1400 (m), 1330 , 1279 (m), 1220 , 1168 , 1084 (m), 1069 (m), 1028 (m), 920 , 887 (ws), 765 (s), 758 (s), 723 (m), 696 (s), 685 (s), 602 (m), 573 (m), 558 (m), 551 (m), 542 (m), 508 (s), 490 (m), 442 (w).IR was prepared by the reaction between equimolar amounts of sodium methano\u00adlate and HL in an methanol medium (20\u2005ml). The mixture was heated with magnetic stirring at 337\u2005K for 10\u2005min. The resulting solution was evaporated and the fine crystalline powder was isolated (yield 83%) and washed with 2-propanol. Dry product NaL was used for the preparation of the complexes. 1H NMR \u03b4 3.46 , 3.48 , 7.26 , 7.53 , 7.6 , 7.77 . 31P NMR \u03b4 54.01.The sodium salt (\u22121): 3068 , 2944 , 2704 , 2660 , 1581 , 1490 (s), 1410 , 1263 , 1271 (m), 1173 , 1165 , 1080 (m), 1031 (m), 891 , 2943 , 2704 , 2660 , 1564 , 1490 (s), 1413 , 1252 , 1243 + \u03bd(C\u2014C)], 1270 (m), 1164 , 1165 , 1082 (m), 1030 (m), 990 , 892 [ws, \u03bd, 763 (s), 747 (s), 720 (m), 678 (s), 682 (s), 564 (m), 547 (m), 534 (m), 502 (s), 485 (m), 427 (w).IR = xUeq(C), where x = 1.5 for methyl H and 1.2 for all other H atoms. A rotating-group model was applied for the methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a02A and B with refined occupancies of 0.50\u2005(3) for both disorder components. The phenyl rings C15\u2013C20, C21\u2013C26 were refined as disordered over two positions with refined occupancies of 0.555\u2005(17) and 0.445\u2005(17), respectively. The bond lenghts C21A\u2014C22A, C22A\u2014C23A, C23A\u2014C24A, C24A\u2014C25A, C25A\u2014C26A, C26A\u2014C21A, C21\u2014C22, C22\u2014C23, C23\u2014C24, C24\u2014C25, C25\u2014C26 and C26\u2014C21 were restrained to have a value of 1.38\u2005(1)\u2005\u00c5 (using a DFIX restraint). The ring carbon atoms C21A, C26A, C25A, C24A, C23A, C22A as well as C21, C22, C23, C24, C25, C26 were restrained to have planar geometries . Anisotropic parameters of all C atoms of disordered rings were restrained to have approximately similar values to within 0.01\u2005\u00c52 (using a SIMU restraint).Phenyl ring C1\u2013C6 was refined as disordered over two positions 3. Satisfactory results (R1 = 5.01%) were obtained modeling disordered C and O atoms, but very large displacement parameters for them were observed. The SQUEEZE procedure per cell. However, the difference in R1 values for the structures with and without the SQUEEZE procedure implemented was rather small (0.5%). In the final refinement, the isolated peaks in the solvent-accessible voids were ignored.During the refinement, several small isolated electron-density peaks were located in solvent-accessible voids that were believed to be solvent mol\u00adecules. The largest residual electron peak accounted to 0.66\u2005e\u2005\u00c510.1107/S2056989017008970/zl2701sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: Click here for additional data file.10.1107/S2056989017008970/zl2701Isup4.cdxSupporting information file. DOI: 10.1107/S2056989017008970/zl2701Isup6.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017008970/zl2701sup5.txthkl-file. DOI: 1556367CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the two indole ring systems are approximately perpendicular to one another, making a dihedral angle of 80.9\u2005(5)\u00b0. In the crystal, pairs of N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into inversion dimers and these are further linked by N\u2014H\u22efO and hydrogen bonds and short Cl\u2014Cl contacts into supra\u00admolecular chains. 29H24Cl2N2O4, the mean planes of the two indole ring systems (r.m.s. deviations = 0.1249 and 0.0075\u2005\u00c5) are approximately perpendic\u00adular to one another, with a dihedral angle of 80.9\u2005(5)\u00b0 between them. The benzene ring is inclined to the mean planes of the two indole ring systems by 76.1\u2005(3) and 78.3\u2005(4)\u00b0. Weak intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions affect the mol\u00adecular conformation. In the crystal, pairs of N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into inversion dimers which are further linked into supra\u00admolecular chains by N\u2014H\u22efO hydrogen bonds and short Cl\u2014Cl contacts.In the title compound, C C11\u2014H11A\u22efCl1 and short Cl2\u22efCl2 contacts [Cl2\u22efCl2 = 3.467\u2005(2)\u2005\u00c5] bridge these chains and form sheets of mol\u00adecules parallel to bis\u00ad(1H-indole-2-carboxyl\u00adate) was dissolved in 20\u2005ml ethanol; commercially available 2,4-di\u00adchloro\u00adbenzalde\u00adhyde was added and the mixture was heated to reflux temperature. Concentrated HCl (0.5\u2005ml) was added and the reaction was left for 1\u2005h. After cooling, the white product was filtered off and washed thoroughly with ethanol. The reaction was monitored with TLC (AcOEt:hexane = 1:3). Colourless block-like crystals of the title compound suitable for X-ray analysis were obtained in 92% yield by slow evaporation of an ethanol solution.Uiso(H) = xUeq, where x = 1.5 for methyl H atoms and 1.2 for all others.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017015730/sj5539sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017015730/sj5539Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017015730/sj5539Isup3.cmlSupporting information file. DOI: 1582719CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Crystal structures of hydrogen-bonded 1:2 compounds of chloranilic acid with 2-pyridone, 3-hy\u00addroxy\u00adpyridine and 4-hyroxypyridine have been determined at 120\u2005K. In each crystal structure, the acid and base mol\u00adecules are linked by short O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds. 5H5NO\u00b7C6H2Cl2O4, (I), bis\u00ad(3-hy\u00addroxy\u00adpyridinium) chloranilate, 2C5H6NO+\u00b7C6Cl2O42\u2212, (II), and bis\u00ad(4-hy\u00addroxy\u00adpyridinium) chloranilate, 2C5H6NO+\u00b7C6Cl2O42\u2212, (III), have been determined at 120\u2005K. In the crystal of (I), the base mol\u00adecule is in the lactam form and no acid\u2013base inter\u00adaction involving H-atom transfer is observed. The acid mol\u00adecule lies on an inversion centre and the asymmetric unit consists of one half-mol\u00adecule of chloranilic acid and one 2-pyridone mol\u00adecule, which are linked via a short O\u2014H\u22efO hydrogen bond. 2-Pyridone mol\u00adecules form a head-to-head dimer via a pair of N\u2014H\u22efO hydrogen bonds, resulting in a tape structure along [201]. In the crystals of (II) and (III), acid\u2013base inter\u00adactions involving H-atom transfer are observed and the divalent cations lie on an inversion centre. The asymmetric unit of (II) consists of one half of a chloranilate anion and one 3-hy\u00addroxy\u00adpyridinium cation, while that of (III) comprises two independent halves of anions and two 4-hy\u00addroxy\u00adpyridinium cations. The primary inter\u00admolecular inter\u00adaction in (II) is a bifurcated O\u2014H\u22ef hydrogen bond between the cation and the anion. The hydrogen-bonded units are further linked via N\u2014H\u22efO hydrogen bonds, forming a layer parallel to the bc plane. In (III), one anion is surrounded by four cations via O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, while the other is surrounded by four cations via N\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds. These inter\u00adactions link the cations and the anions into a layer parallel to (301).The crystal structures of the 1:2 compounds of chloranilic acid with 2-pyridone, 3-hy\u00addroxy\u00adpyridine and 4-hyroxypyridine, namely, bis\u00ad(2-pyridone) chloranilic acid, 2C A weak C\u2014H\u22efCl inter\u00adaction formed between the acid and base mol\u00adecules ii; symmetry code as in Table\u00a02bc plane \u2005\u00c5; symmetry code: (vi) \u2212x\u00a0+\u00a01, y\u00a0\u2212\u00a0z\u00a0+\u00a0In the crystal of (II)ne Fig.\u00a07. Adjacenvia weak C\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions et al., 2016et al., 2010et al., 2016et al., 2009aet al., 2009bA search of the Cambridge Structural Database Uiso(H) = 1.5Ueq(O or N). C-bound H atoms were positioned geometrically (C\u2014H = 0.95\u2005\u00c5) and were treated as riding with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989017013536/lh5855sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989017013536/lh5855Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017013536/lh5855IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989017013536/lh5855IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 1575722, 1575721, 1575720CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The synthesis, characterization and structural analysis of triphenylphosphoniummethylenetrifluoroborate are presented. 19H17BF3P {alternative name: triphen\u00adyl[(tri\u00adfluoro\u00adboran\u00adyl)meth\u00adyl]phosphanium}, was formed by the reaction of tri\u00adphenyl\u00adphosphine with potassium iodo\u00admethyl\u00adtri\u00adfluoro\u00adborate. The mol\u00adecule features a nearly staggered conformation along the P\u2014C bond and a less than staggered conformation along the C\u2014B bond. In the crystal, weak C\u2014H\u22efF hydrogen bonds between the meta-phenyl C\u2014H groups and the tri\u00adfluoro\u00adborate B\u2014F groups form chains of R22(16) rings along [100]. These chains are are further stabilized by weak C\u2014H\u22ef\u03c0 inter\u00adactions. A weak intra\u00admolecular C\u2014H\u22efF hydrogen bond is also observed.The title compound, C Additionally, they may be used to produce organodihaloboranes (RBX2) with boron trifluoride di\u00adethyl\u00adetherate (BF3-OEt2) yields triphen\u00adyl[(tri\u00adfluoro\u00adboran\u00adyl)meth\u00adyl]phosphonium (Ph3PCH2BF3). We have synthesized Ph3PCH2BF3via an alternate route, by reacting tri\u00adphenyl\u00adphosphine (PPh3) with potassium iodo\u00admethyl\u00adtri\u00adfluoro\u00adborate (ICH2BF3K) in 45% yield.Alkyl\u00adtri\u00adphenyl\u00adphospho\u00adnium (PhDatabase survey). Phospho\u00adnium tri\u00adfluoro\u00adborates have been shown to enhance the hydrolytic stability of the RBF3 moiety ring \u00b0] and a less staggered conformation along the C1\u2014B1 bond [F2\u2014B1\u2014C1\u2014P1 torsion angle = 158.3\u2005(2)\u00b0].The mol\u00adecular structure of the title compound is shown in Fig.\u00a01g Table\u00a01. The molet al., 19873sp hybridized. The methyl\u00adene carbon is predominantly 3sp hybridized, but has a distorted tetra\u00adhedral geometry with a P1\u2014C1\u2014B1 angle of 119.7\u2005(2)\u00b0.The B-F bond lengths fall within normal ranges for organotri\u00adfluoro\u00adborate compounds. The methyl\u00adene C\u2014P bond length [1.787\u2005(4)\u2005\u00c5] and the C\u2014B bond length [1.636\u2005(4)\u2005\u00c5] also fall within the normal range for similar compounds and the resulting solid was dissolved in a minimal amount of acetone and the product was precipitated with water and collected by filtration, to afford a white solid X-ray quality crystals were grown by slow diffusion of pentane into a solution of the title compound dissolved in di\u00adchloro\u00admethane.Potassium iodo\u00admethyl\u00adtri\u00adfluoro\u00adborate and tri\u00adphenyl\u00adphosphine were combined in a pressure flask containing a stir bar under nitro\u00adgen, and anhydrous THF (25.0\u2005mL) was added. The flask was sealed and heated to 343\u2005K for 18\u2005h. The reaction was cooled to room temperature and the solvent was removed 1H NMR \u03b4 (ppm): 7.66 , 7.56 , 2.07 . 13C NMR \u03b4 (ppm): 133.7 , 133.5 , 129.6 123.2 (C\u2014B not observed). 11B NMR \u03b4 (ppm): 2.49 . 19F NMR \u03b4 (ppm): \u2212138.9 . FTIR : 3070, 2960, 1587, 1484, 1438, 1146, 1104, 1025, 994, 969, 824, 754, 725, 691, 511, 497.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017009884/lh5846sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017009884/lh5846Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017009884/lh5846Isup3.cmlSupporting information file. DOI: 1560028CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular and crystal structure of the title imidazole derivative is reported. The structure is stabilized by an extensive O\u2014H\u22efN, C\u2014H\u22efO/Cl and C\u2014H\u22ef\u03c0(ring) hydrogen-bonding network. 24H21ClN2O, crystallizes with two unique mol\u00adecules in the asymmetric unit. In each mol\u00adecule, the central imidazole ring is substituted at the 2-, 4- and 5-positions by benzene rings. The 2-substituted ring carries a Cl atom at the 4-position. One of the imidazole N atoms in each mol\u00adecule has a propan-2-ol substituent. In the crystal, a series of O\u2014H\u22efN, C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds, augmented by several C\u2014H\u22ef\u03c0(ring) inter\u00adactions, generate a three-dimensional network of mol\u00adecules stacked along the a-axis direction.The title compound, C Each moO\u22efN23 hydrogen bonds supported by C242\u2014H242\u22efO112 contacts combine with O212\u2014H22O\u22efN13 hydrogen bonds to link alternate type 1 and 2 mol\u00adecules in a head-to-tail fashion, forming C(7) chains along b, Fig.\u00a04C(12) chains along the a-axis direction, Fig.\u00a05Cg5 and C255\u2014H255\u22efCg1 contacts combine with C113\u2014H11D\u22efCg6, C213\u2014H21D\u22efCg2 and two C\u2014H\u22efO hydrogen bonds, Table\u00a01c axis, Fig.\u00a063 across the unit cell. This large void is unexpected as no solvent appeared and the final difference map was reasonably flat . The mol\u00adecules stack in an orderly fashion along each of the three principal crystallographic axes and the voids are clearly visible in views of the overall packing along these directions, see for example Fig.\u00a07O112\u2013H12et al., 2016H-imidazol-2-yl]benzoic acid . All other H atoms were refined using a riding model with d(C\u2014H) = 0.95\u2005\u00c5, Uiso = 1.2Ueq(C) for aromatic, 1.00\u2005\u00c5 for methine and 0.99\u2005\u00c5 for CH2 H atoms, all with Uiso = 1.2Ueq(C) and 0.98\u2005\u00c5, Uiso = 1.5Ueq(C) for CH3 H atoms. Seven reflections with Fo >>> Fc, were omitted from the final refinement cycles.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016019332/hg5480sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989016019332/hg5480Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016019332/hg5480Isup3.cmlSupporting information file. DOI: 1520559CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In (I), the di\u00adthio\u00adcarbamate ligand coordinates the SnIV atom in an asymmetric manner, leading to a highly distorted trigonal\u2013bipyramidal coordination geometry defined by a C3S2 donor set with the weakly bound S atom approximately trans to one of the ipso-C atoms. A similar structure is found in (II), but the di\u00adthio\u00adcarbamate ligand coordinates in an even more asymmetric fashion. The packing in (I) features supra\u00admolecular chains along the c axis sustained by C\u2014H\u22ef\u03c0 inter\u00adactions; chains pack with no directional inter\u00adactions between them. In (II), supra\u00admolecular layers are formed, similarly sustained by C\u2014H\u22ef\u03c0 inter\u00adactions; these stack along the b axis. An analysis of the Hirshfeld surfaces for (I) and (II) confirms the presence of the C\u2014H\u22ef\u03c0 inter\u00adactions but also reveals the overall dominance of H\u22efH contacts in the respective crystals.The crystal and mol\u00adecular structures of two tri\u00adphenyl\u00adtin di\u00adthio\u00adcarbamates, [Sn(C RSn(S2CNRR\u2032)n4\u2013 where R, R\u2032 = alkyl, aryl, most relate to their biological activities and their usefulness as mol\u00adecular, single-source precursors for the formation of tin sulfide nanoparticles 3Sn[S2CN(Ben)CH2CH2Ph] (I)6H5)3Sn[S2CN(CH2CH2OMe)2] (II)Among the varied motivations for investigating organotin di\u00adthio\u00adcarbamate compounds, long and Sn\u2014Sshort bond lengths, of 0.42\u2005\u00c5. This asymmetry is reflected in the relatively large disparity in the associated C\u2014S bond lengths with the bond involving the tightly bound S1 atom being significantly longer than the bond involving the S2 atom, Table\u00a01i.e. from a narrow 92.98\u2005(4)\u00b0 for S1\u2014Sn\u2014C17 to a wide 124.31\u2005(4)\u00b0 for S1\u2014Sn\u2014C29. The wide angle is due to the close approach to the tin atom of S2. Further, the Sn\u2014C17 bond length is systematically longer than the other Sn\u2014C bond lengths, an observation ascribed to the C17 atom being approximately trans to the incoming S2 atom, Table\u00a013S2 donor set. The geometry is not ideal with the value of \u03c4 of 0.57, cf. \u03c4 values of 0.0 and 1.0 for ideal square\u2013pyramidal and trigonal\u2013bipyramidal geometries, respectively cf. 0.06\u2005\u00c5 for (I)trans to the S2 atom, is the longest of all six Sn\u2014C bonds in (I)i.e. 90.94\u2005(4)\u00b0 for S1\u2014Sn\u2014C14 to 119.54\u2005(5)\u00b0 for C8\u2014Sn\u2014C20, is slightly narrower at less than 30\u00b0. The value of \u03c4 computes to 0.58, i.e. virtually identical to that in (I)The mol\u00adecular structure of (II)PLATON ac plane, Fig.\u00a04a. The layers stack along the b axis without directional inter\u00adactions between them, Fig.\u00a04b.Even though there are oxygen atoms in the mol\u00adecule of (II)Crystal Explorer di and de from the Hirshfeld surface to the nearest atom inside and outside, respectively, enable the analysis of the inter\u00admolecular inter\u00adactions through the mapping of dnorm. The combination of de and di in the form of two-dimensional fingerprint plots dnorm (not shown). The curvature of the Hirshfeld surfaces around the phenyl rings participating as acceptors in the C\u2014H\u22ef\u03c0 contacts determine the strength of these inter\u00adactions in the crystal packing. In the structure of (I)i.e. with the H13 and H23 hydrogen atoms. The other C\u2014H\u22ef\u03c0 contact involves methyl-H7C atom as the donor and phenyl (C8\u2013C13) ring as the acceptor. The shape-indexed Hirshfeld surfaces highlighting the C\u2014H\u22ef\u03c0 contacts are shown in Fig.\u00a07The different shapes of Hirshfeld surfaces for mol\u00adecules (I)et al., 2007a\u2013d, respectively; their relative contributions are summarized in Table\u00a04de, di) ranges, i.e. between 1.2 and 2.6\u2005\u00c5, the densities and the areas of their distributions are different. It is evident from the data in Table\u00a04b that these contacts make the most significant contribution to the Hirshfeld surfaces of both (I)b), all the points are situated at the distances greater than or equal to their van der Waals separations i.e. 2 x 1.2\u2005\u00c5, hence there is no propensity to form such inter\u00admolecular contacts. The peak at distances slightly less than van der Waals separations in the fingerprint plot for (II)A\u22efH5Ai = 2.36\u2005\u00c5; symmetry code: (i) \u2212x, 2\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z]. In the fingerprint plot delineated into C\u22efH/H\u22efC contacts for (I)c, the 32.9% contribution to the Hirshfeld surface and the symmetrical distribution of points showing bending of the pattern at (de + di)min \u223c2.8\u2005\u00c5 is the result of short inter\u00adatomic C\u22efH/H\u22efC contacts . In the structure of (II)i.e. 24.4%, an observation ascribed to the presence of only C\u2014H\u22ef\u03c0 contacts in the mol\u00adecular packing, with no other short inter-atomic contacts. The negligible contribution from C\u22efC contacts to the Hirshfeld surfaces indicate that despite the presence of three Sn-bound phenyl rings in the structures of both (I)d, show that crowded geometries around the tin atoms prevent the sulfur atoms from forming such inter\u00admolecular contacts although these contacts have significant contributions to their respective Hirshfeld surfaces, Table\u00a04et al., 2016The overall two-dimensional fingerprint plots for (I)et al., 2016R/R\u2032 substituents. For example, the \u2212S2CN(Ben)CH2CH2Ph anion in (I)2CN(Ben)CH2CH2Ph]2 2 anion, as in (II)et al., 2009et al., 2010According to a search of the Cambridge Structural Database features the shortest Sn\u2014S bond length of the series at 2.429\u2005(3)\u2005\u00c5 of 0.74\u2005\u00c5, is found in the structure of Ph3Sn(4-nitro\u00adphenyl\u00adpiperazine-1-di\u00adthio\u00adcarbamate) , having \u0394(Sn\u2014S) of 0.42\u2005\u00c5 Reflecting the inter\u00adest in organotin di\u00adthio\u00adcarbamates, there are approximately 40 examples of tri\u00adphenyl\u00adtin di\u00adthio\u00adcarbamate structures in the CSD, all of which present the same basic structural motif as described herein for (I)N-Benzyl-2-phenyl\u00adethyl\u00adamine (2\u2005mmol) dissolved in ethanol (10\u2005ml) was stirred for 30\u2005min in an ice-bath. 25% ammonia (1\u20132\u2005ml) was added to generate a basic solution. After that, a cold ethano\u00adlic solution of carbon di\u00adsulfide (2\u2005mmol) was added to the solution followed by stirring for about 2\u2005h. Then, tri\u00adphenyl\u00adtin(IV) chloride (2\u2005mmol) dissolved in ethanol (30\u2005ml) was added drop wise into the solution followed by further stirring for 2\u2005h. The precipitate that formed was filtered off and washed with cold ethanol a few times to remove impurities. Finally, the precipitate was dried in a desiccator. Recrystallization was achieved by dissolv\u00ading the compound in a chloro\u00adform and ethanol mixture (1:1 v/v): this solution was allowed to slowly evaporate at room temperature yielding colourless slabs of (I)34H31NS2Sn requires: C, 64.2; H, 4.9; N, 2.2; S, 10.1. IR (cm\u22121): 1476 \u03bd(C\u2014N), 1021 \u03bd(C\u2014S), 502 \u03bd(Sn\u2014C), 448 \u03bd(Sn\u2014S). 1H NMR (CDCl3): 7.44\u20137.86 , 7.16\u20137.39 , 5.03 , 3.96 , 3.04 . 13C{1H} NMR (CDCl3): \u03b4 197.8 (S2C), 126.7\u2013142.3 (Ar), 59.8 (CH2Ben), 56.4 (NCH2CH2), 32.8 (NCH2CH2). 119Sn{1H} NMR (CDCl3): \u2212180.2.Synthesis of (I)N-benzyl-2-phenyl\u00adethyl\u00adamine. Recrystallization was from chloro\u00adform solution to yield colourless slabs. M.p.: 366\u2013367\u2005K. Yield: 89%. Analysis: found C, 54.4; H, 4.4; N, 2.9; S, 12.1. C25H29NO2S2 Sn requires: C, 53.8; H, 5.2; N, 2.5; S, 11.5. IR (cm\u22121): 1470 \u03bd(C\u2014N), 994 \u03bd(C\u2014S), 559 \u03bd(Sn\u2014C), 425 \u03bd(Sn\u2014S). 1H NMR (CDCl3): 7.40\u20137.74 , 4.13 , 3.72 , 3.35 . 13C{1H} NMR (CDCl3): \u03b4 197.3 (S2C), 128.6\u2013142.4 (Ar), 70.0 (OCH2), 59.0 (NCH2), 57.1 (CH3). 119Sn{1H} NMR (CDCl3): \u2212185.0.Compound (II)Uiso(H) set to 1.2\u20131.5Ueq(C). In the refinement of (II)Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989016014985/hb7618sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989016014985/hb7618Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016014985/hb7618IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1505733, 1505732CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E. The packing features amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds, leading to centrosymmetric aggregates, as well as C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions, which significantly influence the packing.An intra\u00admolecular amine-N\u2014H\u22efN(imine) hydrogen bond is found in the disubstituted amino\u00adurea residue; the conformations about the imine and ethyl\u00adene double bonds are 23H21N3O2, is constructed about an almost planar disubstituted amino\u00adurea residue (r.m.s. deviation = 0.0201\u2005\u00c5), which features an intra\u00admolecular amine-N\u2014H\u22efN(imine) hydrogen bond. In the \u2018all-trans\u2019 chain connecting this to the terminal meth\u00adoxy\u00adbenzene residue, the conformation about each of the imine and ethyl\u00adene double bonds is E. In the crystal, amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds connect centrosymmetrically related mol\u00adecules into dimeric aggregates, which also incorporate ethyl\u00adene-C\u2014H\u22efO(amide) inter\u00adactions. The dimers are linked by amine\u2013phenyl-C\u2014H\u22ef\u03c0(imine\u2013phen\u00adyl) and meth\u00adoxy\u00adbenzene-C\u2014H\u22ef\u03c0(amine\u2013phen\u00adyl) inter\u00adactions to generate a three-dimensional network. The importance of C\u2014H\u22ef\u03c0 inter\u00adactions in the mol\u00adecular packing is reflected in the relatively high contributions made by C\u22efH/H\u22efC contacts to the Hirshfeld surface, i.e. 31.6%.The title compound, C These compounds have attracted much attention due to their diverse pharmacological and biological activities 3O = 0.0201\u2005\u00c5), owing in part to an intra\u00admolecular amine-N\u2014H\u22efN(imine) hydrogen bond, Table\u00a013O plane, forming a dihedral angle of 46.88\u2005(4)\u00b0. The imine/ethyl\u00adene sequence of bonds, i.e. N3=C8\u2014C9=C10\u2014C11, has an all-trans conformation but the N3\u2014C8\u2014C9\u2014C10 and C8\u2014C9\u2014C10\u2014C11 torsion angles of 154.62\u2005(12) and \u2212169.19\u2005(11)\u00b0, respectively, indicate some twisting in this residue, especially about the C8\u2014C9 bond; the conformation about each of the double bonds is E. The imine-bound phenyl ring forms a dihedral angle of 63.30\u2005(7)\u00b0 with the C4N atoms of the imine/ethyl\u00adene sequence, and the corresponding angle for the terminal meth\u00adoxy\u00adbenzene ring is significantly less, at 8.29\u2005(13)\u00b0. The meth\u00adoxy group is twisted out of the plane of the ring to which it is connected as seen in the value of the C17\u2014O18\u2014C14\u2014C15 torsion angle of 15.55\u2005(17)\u00b0.The mol\u00adecular structure of (I)2, Table\u00a01a, as well as meth\u00adoxy-C\u2014H\u22ef\u03c0(amine-phen\u00adyl) contacts, Table\u00a01via amine-phenyl-C\u2014H\u22ef\u03c0(imine-phen\u00adyl) and meth\u00adoxy-benzene-C\u2014H\u22ef\u03c0(amine-phen\u00adyl) inter\u00adactions, Fig.\u00a02b.The most notable feature of the mol\u00adecular packing of (I)et al., 2017N and carbonyl-O1 atoms on the Hirshfeld surface mapped over dnorm in Fig.\u00a03a. The appearance of diminutive red spots near the N3 and C17 atoms, Fig.\u00a03a, and the tiny faint-red spots near the C9 and H82 atoms in Fig.\u00a03b, indicate the influence of short inter\u00adatomic N3\u22efC17 and C9\u22efH82 contacts, Table\u00a02a, the concave region around the imine-phenyl ring on one side and the biconcave region around the amine-phenyl ring indicate their involvement in one and two C\u2014H\u22ef\u03c0 contacts, respectively. The short inter\u00adatomic O\u22efH/H\u22efO contacts s Table\u00a03 as well a, and those delineated into H\u22efH, C\u22efH/H\u22efC, O\u22efH/H\u22efO and N\u22efH/H\u22efN contacts c, which is due to the presence of a significant number of C\u2014H\u22ef\u03c0 inter\u00adactions involving the imine- and amine-phenyl rings, as well as short inter\u00adatomic C\u22efH/H\u22efC contacts, Table\u00a03c. The pair of forceps-like long tips at de + di = 2.1\u2005\u00c5 in the fingerprint plot delineated into O\u22efH/H\u22efO contacts, Fig.\u00a06d, reflect the presence the N\u2014H\u22efO hydrogen bond; the pair of spikes corresponding to the C\u2014H\u22efO contacts and the points related to short inter\u00adatomic O\u22efH/H\u22efO contacts, Table\u00a02e, as their inter\u00adatomic distances are greater than their van der Waals separations, they do not make a specific contribution to the mol\u00adecular packing. The participation of the methyl-C17 atom in two close inter\u00adatomic contacts, Table\u00a02A and H17B atoms so are not surface contacts. Finally, the small contributions from other inter\u00adatomic contacts summarized in Table\u00a03The C\u22efH/H\u22efC contacts in the crystal make the second largest contribution, et al., 2016et al., 2005E-conformations about the two analogous double bonds in the mol\u00adecule. However, there is considerable twisting about the equivalent bonds to C8\u2014C9 in (I)i.e. the N\u2014C\u2014C\u2014C torsion angles are 130.3\u2005(6) and 136.0\u2005(6)\u00b0, cf. 154.62\u2005(12)\u00b0 in (I)The title compound was prepared from the de\u00adhydrogenation reaction of 4-phenyl\u00adsemicarbazide and 4-meth\u00adoxy\u00adchalcone. A search of the Cambridge Structural Database , m.p. 407\u2005K. IR (cm\u22121): 3336 \u03bd(N\u2014H), 1679 \u03bd(C=O), 1526 \u03bd(C=N), 1242 \u03bd(C\u2014N), 1025 \u03bd(C=S). MS (m/z): 371.25 [M+1]+.Analytical grade reagents were used as procured without further purification. 4-Phenyl\u00adsemicarbazide and 4-meth\u00adoxy\u00adchalcone were dissolved separately in hot absolute ethanol (30\u2005ml) and mixed with stirring. A few drops of concentrated hydro\u00adchloric acid were added as a catalyst. The reaction mixture was heated and stirred for about 20\u2005min., then stirred for a further 30\u2005min. at room temperature. The resulting yellow precipitate was filtered, washed with cold ethanol and dried Uiso(H) set to 1.2\u20131.5Ueq(C). The nitro\u00adgen-bound H atoms were located in a difference-Fourier map but were refined with a distance restraint of N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989017014128/hb7709sup1.cifCrystal structure: contains datablock(s) . DOI: 10.1107/S2056989017014128/hb7709Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017014128/hb7709Isup3.cmlSupporting information file. DOI: 1577439CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "IV atom in [Sn(C7H6F)2Cl2(C2H6OS)2] is located on a centre of inversion so the resulting donor C2Cl2O2 donor set is all-trans. The three-dimensional mol\u00adecular packing is sustained by C\u2014H\u22efF, C\u2014H\u22efCl and C\u2014H\u22ef\u03c0 inter\u00adactions.The octa\u00adhedrally coordinated Sn IV atom in the title diorganotin compound, [Sn(C7H6F)2Cl2(C2H6OS)2], is located on a centre of inversion, resulting in the C2Cl2O2 donor set having an all-trans disposition of like atoms. The coordination geometry approximates an octa\u00adhedron. The crystal features C\u2014H\u22efF, C\u2014H\u22efCl and C\u2014H\u22ef\u03c0 inter\u00adactions, giving rise to a three-dimensional network. The respective influences of the Cl\u22efH/H\u22efCl and F\u22efH/H\u22efF contacts to the mol\u00adecular packing are clearly evident from the analysis of the Hirshfeld surface.The Sn The SnIV atom is coordinated by monodentate ligands, i.e. chloride, sulfoxide-O and methyl\u00adene-C atoms. From symmetry, each donor is trans to a like atom resulting in an all-trans-C2Cl2O2 donor set about the SnIV atom. The donor set defines a distorted octa\u00adhedral geometry owing, in part, to the disparate Sn\u2014donor atom bond lengths, Table\u00a01IV atom differ relatively little from the ideal octa\u00adhedral angles with the maximum deviation of ca 6\u00b0 noted for the C1\u2014Sn\u2014O1 angle, Table\u00a01The mol\u00adecular structure of (I)bc plane, Fig.\u00a02a. Layers are connected along the a axis by phenyl-C3\u2014H\u22efF1 and methyl-C9\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adactions to consolidate the mol\u00adecular packing, Fig.\u00a02b.The mol\u00adecular packing in (I)et al., 2016dnorm in Fig.\u00a03B and a pair near Cl1 in Fig.\u00a03dnorm- and shape-index-mapped Hirshfeld surfaces highlighting the various C\u2014H\u22efCl, C\u2014H\u22efF and C\u2014H\u22ef\u03c0 inter\u00adactions are illustrated in Fig.\u00a05a\u2013c, respectively.The Hirshfeld surface analysis on the structure of (I)et al., 2007a\u2013f, respectively, and their relative contributions to the Hirshfeld surfaces are summarized in Table\u00a04b, that although these contacts have the greatest contribution, i.e. 45.7%, to the Hirshfeld surface, the dispersion forces acting between them keep these atoms at the distances greater than the sum of their van der Waals radii, hence they do not contribute significantly to the mol\u00adecular packing. The comparatively greater contribution of F\u22efH/H\u22efF contacts to the Hirshfeld surface cf. Cl\u22efH/H\u22efCl contacts, Table\u00a04c. The forceps-like distribution of points in the plot with tips at de + di \u223c2.8\u2005\u00c5 result from the bifurcated C\u2014H\u22efCl inter\u00adactions, and points at positions less than the sum of their van der Waals radii are ascribed to the short inter\u00adatomic Cl\u22efH/H\u22efCl contacts, the green appearance due to high density of inter\u00adactions. Similarly, a pair of short spikes at de + di \u223c2.5\u2005\u00c5 in the fingerprint plot delineated into F\u22efH/H\u22efF contacts, Fig.\u00a06d, are indicative of inter\u00admolecular C\u2014H\u22efF inter\u00adactions with the short inter\u00adatomic F\u22efH/H\u22efF contacts merged within the fingerprint plot. It is important to note from the fingerprint plot delineated into C\u22efH/H\u22efC contacts, Fig.\u00a06e, that even though their inter\u00adatomic distances are equal to or greater than the sum of their van der Waals radii, i.e. 2.9\u2005\u00c5, the 12.8% contribution from these to the Hirshfeld surfaces are indicative of the presence of C\u2014H\u22ef\u03c0 inter\u00adactions in the structure. This is also justified from the presence of short inter\u00adatomic C\u22efC contacts, Fig.\u00a05c and Table\u00a03f, and the small contributions from the other contacts listed in Table\u00a02The overall two-dimensional fingerprint plot and those delineated into H\u22efH, Cl\u22efH/H\u22efCl, F\u22efH/H\u22efF, C\u22efH/H\u22efC and O\u22efH/H\u22efO contacts (McKinnon R2SnX2(DMSO)2 in the crystallographic literature 2 compound tin dichloride was prepared in accordance with the literature method and tri\u00adethyl\u00adamine in ethyl acetate (25\u2005ml) was added to di(p-fluoro\u00adbenz\u00adyl)tin dichloride in ethyl acetate (10\u2005ml). The resulting mixture was stirred and refluxed for 4\u2005h. The filtrate was evaporated until a dark-orange precipitate was obtained. The precipitate was dissolved in DMSO-d6 solution in a NMR tube for 1H NMR spectroscopic characterization. After the analysis, the tube was set aside for a month and colourless crystals of (I)\u22121): 1595(m) \u03bd(C=C), 1504(s) \u03bd(S=O), 1161(m), 578(w), 508(m) \u03bd(Sn\u2014O). 1H NMR (in CDCl3): 6.90\u20137.11, 7.35\u20137.40 , 3.11 , 2.17 .Uiso(H) set to 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989017005072/wm5381sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017005072/wm5381Isup2.hklStructure factors: contains datablock(s) I. DOI: 1541712CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular and crystal structures of the centrosymmetric naphthoxazine derivative is reported. In the absence of hydrogen-bonding and \u03c0\u2013\u03c0 stacking inter\u00adactions, the crystal structure is stabilized by short C\u2014H\u22ef\u03c0 contacts. 26H24N2O2, the oxazine moiety is fused to a naphthalene ring system. The asymmetric unit consists of one half of the mol\u00adecule, which lies about an inversion centre. The C atoms of the ethyl\u00adene spacer group adopt an anti\u00adperiplanar arrangement. The oxazine ring adopts a half-chair conformation. In the crystal, supra\u00admolecular chains running along the b axis are formed via short C\u2014H\u22ef\u03c0 contacts. The crystal studied was a non-merohedral twin with a fractional contribution of 0.168\u2005(2) of the minor twin component.In the title compound, C Evaporation at room temperature afforded the title compound in 73% yield after recrystallization.The oxazine moiety is well known as a building block for high-performance phenolic resins, which are of great inter\u00adest in industry due to their superior mechanical and physical properties together with unusually high thermal resistance \u2005\u00c5, \u03b8 = 51.5\u2005(4)\u00b0, \u03c6 = 86.6\u2005(4)\u00b0, and the ethyl\u00adene spacer group adopts an anti\u00adperiplanar arrangement as observed in 3,3\u2032-bis\u00ad 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z]. However, unlike the related structures, which crystallized in monoclinic space groups with one mol\u00adecule in the asymmetric unit .In general terms, the structure of the title compound Fig.\u00a01 is similet al., 2007et al., 2015The aromatic C\u2014C bonds of naphthalene ring system have a narrow range of distances [from 1.365\u2005(5) to 1.431\u2005(4)\u2005\u00c5]. The central C5\u2014C10 bond at 1.415\u2005(4)\u2005\u00c5 is, however, shorter by 0.014\u2005\u00c5 than those in related structures set to 1.2Ueq of the parent atom. The crystal was a non-merohedral twin with a fractional contribution of 0.168\u2005(2) of the minor twin component.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017006673/sj5529sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017006673/sj5529Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017006673/sj5529Isup3.cmlSupporting information file. DOI: 1547729CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II complex, the divalent Ni ion occupies a crystallographically imposed centre of symmetry and is coordinated by two O atoms from the carboxyl\u00adate groups of two 2,4,6-tri\u00admethyl\u00adbenzoate ligands, two N atoms from the pyridyl groups of two isonicotinamide ligands and two water mol\u00adecules in a slightly distorted octa\u00adhedral geometry. Hirshfeld surface analysis indicates that the most important contributions for the crystal packing are from H\u22efH (59.8%), O\u22efH/H\u22efO (20.2%) and C\u22efH/H\u22efC (13.7%) inter\u00adactions.In the title Ni II complex, [Ni(C10H11O2)2(C6H6N2O)2(H2O)2]\u00b72H2O, the divalent Ni ion occupies a crystallographically imposed centre of symmetry and is coordinated by two O atoms from the carboxyl\u00adate groups of two 2,4,6-tri\u00admethyl\u00adbenzoate (TMB) ligands [Ni\u2014O = 2.0438\u2005(12)\u2005\u00c5], two N atoms from the pyridyl groups of two isonicotinamide (INA) ligands [Ni\u2014N = 2.1506\u2005(15)\u2005\u00c5] and two water mol\u00adecules [Ni\u2014O = 2.0438\u2005(12)\u2005\u00c5] in a slightly distorted octa\u00adhedral geometry. The coordinating water mol\u00adecules are hydrogen bonded to the non-coordinating carboxyl\u00adate O atom of the TMB ligand [O\u22efO = 2.593\u2005(3)\u2005\u00c5], enclosing an S(6) hydrogen-bonding motif. Two solvent water mol\u00adecules are also present in the formula unit. In the crystal, a network of inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds link the complexes into a three-dimensional array. Hirshfeld surface analysis indicates that the most important contributions for the crystal packing are from H\u22efH (59.8%), O\u22efH/H\u22efO (20.2%) and C\u22efH/H\u22efC (13.7%) inter\u00adactions.In the title Ni A deficiency in this vitamin leads to loss of copper from the body, giving rise to a condition known as pellagra disease. Victims of pellagra show unusually high serum and urinary copper levels carboxyl\u00adates are diverse and include one-dimensional (Guseinov 7H4ClO2)2(C6H6N2O)2(H2O)2] \u00b72(H2O) \u00b72H2O, and report its mol\u00adecular and crystal structures, along with a Hirshfeld surface analysis.The structures of a number of mononuclear complexes of divalent transition-metal ions with both nicotinamide (NA) and benzoic acid derivatives as ligands have been previously reported and include hydrogen bonds (Table\u00a01The asymmetric unit of the mononuclear title compound (I)er Fig.\u00a01. In the z] Fig.\u00a01. The nons Table\u00a01. Intra\u00adms Table\u00a01 link H as Table\u00a01.A (C2\u2013C7), ring by 78.80\u2005(14)\u00b0, while the benzene and pyridine, B (N1/C11\u2013C15), rings are oriented at a dihedral angle of 24.33\u2005(6)\u00b0.The near equalities of the C1\u2014O1 [1.242\u2005(2)\u2005\u00c5] and C1\u2014O2 [1.260\u2005(2)\u2005\u00c5] bonds in the carboxyl\u00adate groups indicate delocalized bonding arrangements, rather than localized single and double bonds. The O2\u2014C1\u2014O1 bond angle [124.52\u2005(17)\u00b0] is comparable the corresponding values of 124.4\u2005(2)\u00b0 in (II), 124.67\u2005(14)\u00b0 in (III), 124.22\u2005(11)\u00b0 in (IV), 125.71\u2005(10)\u00b0 in (V), 126.0\u2005(3)\u00b0 in (VI) and 120.47\u2005(15) and 123.17\u2005(15)\u00b0 in (VII), where the benzoate ions also coordinate the metal atoms monodentately. The Ni1 atom lies 0.3523\u2005(1)\u2005\u00c5 below the planar (O1/O2/C1) carboxyl\u00adate group. In the TMB anion, the carboxyl\u00adate group is twisted away from the attached benzene, coordW\u22efOnoncoordW, O\u2014HnoncoordW\u22efOc, N\u2014HINA\u22efOnoncoordW and N\u2014HINA\u22efOINA (INA = isonicotinamide and noncoordW = non-coordinating water) hydrogen bonds . The presence of these inter\u00adactions may also be shown by the Hirshfeld surface mapped as a function of curvedness in H2O (50\u2005ml) and isonicotinamide in H2O (25\u2005ml) with sodium 2,4,6-tri\u00admethyl\u00adbenzoate in H2O (150\u2005ml) at room temperature. The mixture was set aside to crystallize at ambient temperature for nine weeks and gave green single crystals . Combustion analysis: found; C, 54.70, H, 6.24; N, 8.13%. Calculated: C32H42N4NiO10 C, 54.80; H, 6.04; N, 7.99%. FT\u2013IR: 3354, 3197, 2235, 1949, 1855, 1698, 1934, 1612, 1557, 1415, 1226, 1182, 1148, 1115, 1096, 1066, 1041, 1017, 985, 885, 855, 792, 772, 747, 682, 660, 638, 615, 520, 443\u2005cm \u22121.The title compound was prepared by mixing solutions of NiSO2 groups and water mol\u00adecules were located in difference Fourier maps and refined freely. The C-bound H atoms were positioned geometrically with C\u2014H = 0.93 and 0.96\u2005\u00c5 for aromatic and methyl H atoms, respectively, and constrained to ride on their parent atoms, with Uiso(H) = k \u00d7 Ueq(C), where k = 1.5 for methyl H atoms and k = 1.2 for aromatic H atoms. The maximum and minimum residual density peaks were found at 0.83 and 0.78\u2005\u00c5 from atoms O1 and O4, respectively.The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0210.1107/S205698901701060X/cq2020sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901701060X/cq2020Isup2.hklStructure factors: contains datablock(s) I. DOI: 1562879CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Inter\u00admolecular N\u2014H\u22efO and C \u2014H\u22efO hydrogen bonds connect centrosymmetrically related mol\u00adecules into dimers, forming rings of a axis into a columnar arrangement.The title Schiff base mol\u00adecule displays a 16H16N4O5S\u00b7C3H7NO, displays a trans conformation with respect to the C=N double bond. The C\u2014N and N\u2014N bonds are relatively short compared to their normal bond lengths, indicating some degree of delocalization in the mol\u00adecule. The mol\u00adecule is bent at the S atom, with an S\u2014N\u2014C\u2014C torsion angle of 164.48\u2005(11)\u00b0. The dihedral angle between the two aromatic rings is 84.594\u2005(7)\u00b0. Inter\u00admolecular N\u2014H\u22efO and C \u2014H\u22efO hydrogen bonds connect centrosymmetrically related mol\u00adecules into dimers forming rings of R33(11) and R22(10) graph-set motif stacked along the a axis into a columnar arrangement. The mol\u00adecular columns are further linked into a three-dimensional network by C\u2014H\u22ef\u03c0 inter\u00adactions.The mol\u00adecule of the title Schiff base compound, C N-(p-toluene\u00adsulfon\u00adyl)amino acids have been studied extensively for their biological and medicinal activities -4-methyl-N-{2-[2-(4-nitro\u00adbenzyl\u00adidene)hydrazin-1-yl]-2-oxoeth\u00adyl}benzene\u00adsulfonamide N,N-di\u00admethyl\u00adformamide monosolvate.Hydrazones possess a wide variety of biological activities which include anti-inflammatory, analgesic, anti\u00adconvulsant, anti\u00adtuberculous, anti\u00adtumor, anti-HIV and anti\u00admicrobial activity. Hydrazones and their derivatives which can be prepared easily are stable and crystalline in nature. These characteristics have made them suitable compounds in recent times for drug design, ligands for metal complexes and for heterocyclic synthesis. Thus, hydrazones derived from syn to each other. The C8\u2014O3 and C9\u2014N3 bond lengths of 1.219\u2005(2) and 1.274\u2005(2)\u2005\u00c5, respectively, confirm their significant double-bond characters. Further, the C8\u2014N2 and N2\u2014N3 bond lengths of 1.354\u2005(2) and 1.3723\u2005(18)\u2005\u00c5, respectively, also indicate a significant delocalization of \u03c0-electron density over the hydrazone portion of the mol\u00adecule. The mol\u00adecule is bent at the S atom, with an S1\u2014N1\u2014C7\u2014C8 torsion angle of 164.48\u2005(11)\u00b0. The sulfonamide bond exists in a synclinal conformation, with a C\u2014S\u2014N\u2014C torsion angle of \u221278.2\u2005(1)\u00b0, which is the most preferred conformation for aromatic sulfon\u00adamides \u00b0 and C6\u2014C1\u2014S1\u2014N1 = 98.87\u2005(16)\u00b0, while that of the hydrazone group with the attached 4-nitrobenzene ring is given by the torsion angles C11\u2014C10\u2014C9\u2014N3 = 1.6\u2005(2)\u00b0 and C15\u2014C10\u2014C9\u2014N3 = \u2212177.27\u2005(15)\u00b0, respectively. The dihedral angle between the C1\u2013C6 sulfonyl benzene ring and the mean plane through the SO2\u2014NH\u2014CH2\u2014CO segment is 81.452\u2005(6)\u00b0, while that between the C10\u2013C15 benzene ring and the plane through the C9\u2014N3\u2014N2\u2014CO group is 4.296\u2005(10)\u00b0. The dihedral angle between the two aromatic rings is 84.594\u2005(7)\u00b0. The central part of the title compound, between atoms N1 and C9, is nearly planar with an extended chain conformation. The two benzene rings, i.e. C1\u2013C6 and C10\u2013C15, are inclined to the mean plane of the central spacer unit by 85.59\u2005(8) and 4.35\u2005(8)\u00b0, respectively.The title compound crystallizes as a di\u00admethyl\u00adformamide (DMF) monosolvate with one mol\u00adecule each of the Schiff base and solvent in the asymmetric unit Fig.\u00a01, and twoa axis by C\u2014H\u22efO hydrogen bonds involving aromatic C3 and sulfonyl O3 atoms -N-{2-[2-(4-methyl\u00adbenzyl\u00adidene)hydrazin-1-yl]-2-oxoeth\u00adyl}-p-toluene\u00adsulfonamide, (II) E)-N-{2-[2-(4-Nitro\u00adbenzyl\u00adidene)hydrazine-1-yl]-2-oxoeth\u00adyl}-4-methyl\u00adbenzene\u00adsulfonamide N,N-di\u00admethyl\u00adformamide mono\u00adsolvate was prepared as follows: p-toluene\u00adsulfonyl chloride (0.01\u2005mol) was added to glycine (0.02\u2005mol) dissolved in an aqueous solution of potassium carbonate . The reaction mixture was stirred at 373\u2005K for 6\u2005h, left overnight at room temperature, then filtered and treated with dilute hydro\u00adchloric acid. The solid N-(4-methyl\u00adbenzene\u00adsulfon\u00adyl)glycine (L1) obtained was crystallized from aqueous ethanol. Sulfuric acid (0.5\u2005ml) was added to L1 (0.02\u2005mol) dissolved in ethanol (30\u2005ml) and the mixture was refluxed. The reaction mixture was monitored by thin-layer chromatography (TLC) at regular inter\u00advals. After completion of the reaction, the reaction mixture was concentrated to remove excess ethanol. The product, N-(4-methyl\u00adbenzene\u00adsulfon\u00adyl)glycine ethyl ester (L2), was poured into water, neutralized with sodium bicarbonate and recrystallized from acetone. Pure L2 (0.01\u2005mol) was then added in small portions to a stirred solution of 99% hydrazine hydrate (10\u2005ml) in 30\u2005ml ethanol and the mixture was refluxed for 6\u2005h. After cooling to room temperature, the resulting precipitate was filtered, washed with cold water and dried to obtain N-(4-methyl\u00adbenzene\u00adsulfon\u00adyl)glycinyl hydrazide (L3). A mixture of L3 (0.01\u2005mol) and p-nitro\u00adbenzaldehyde (0.01\u2005mol) in anhydrous methanol (30\u2005ml) and two drops of glacial acetic acid was refluxed for 8\u2005h. After cooling, the precipitated (E)-N-{2-[2-(4-nitro\u00adben\u00adzyl\u00adidene)hydrazine-1-yl]-2-oxoeth\u00adyl}-4-methyl\u00adbenzene\u00adsul\u00adfon\u00adamide was collected by vacuum filtration, washed with cold methanol, dried and recrystallized to constant melting point from methanol (522\u2013523\u2005K). The purity of the compound was checked by TLC and characterized by its IR spectrum. The characteristic absorptions observed are 3236.6, 1687.7, 1587.4, 1338.6 and 1163.1\u2005cm\u22121 for the stretching bands of N\u2014H, C=O, C=N, S=O asymmetric and S=O symmetric, respectively. The characteristic 1H and 13C NMR specta of the title compound are as follows. 1H NMR , 3.61, 4.10 , 7.36\u20137.39 , 7.72\u20137.74 , 7.86 , 8.23\u20138.27 , 7.93 , 8.02 , 11.74 . 13C NMR : \u03b4 20.91, 43.20, 44.55, 123.94, 126.60, 127.81, 129.48, 137.48, 140.24, 141.40, 142.64, 144.62, 147.73, 164.64, 169.44. Prism-like colourless single crystals of the title compound employed in the X-ray diffraction study were grown from a DMF solution by slow evaporation of the solvent. or 1.5Ueq(C) for methyl H atoms. A rotating model was used for the methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017014669/rz5222sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017014669/rz5222Isup2.hklStructure factors: contains datablock(s) I. DOI: 1433602CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III(CO)(C(dppm)2-\u03ba3P,C,P\u2032)ClH]Cl and Cl2 have been determined. Both complexes show a slightly distorted octa\u00adhedral coordinated IrIII centre. The PCP pincer ligand system is arranged in a meridional manner.The crystal structures of [Ir et al. meth\u00adyl]di\u00adphenyl\u00adphosphanyl\u00adidene}methane-\u03ba3P,C,P\u2032)carbonyl\u00adchlorido\u00adhydridoiridium(III) chloride di\u00adchloro\u00admethane tris\u00adolvate, Cl\u00b73CH2Cl2 (1) and the closely related (bis\u00ad{[(di\u00adphenyl\u00adphosphan\u00adyl)meth\u00adyl]di\u00adphenyl\u00adphosphanyl\u00adidene}methanide(1+)-\u03ba3P,C,P\u2032)carbonyl\u00adchlorido\u00adhy\u00addridoirid\u00adium(III) dichloride\u2013hydro\u00adchloric acid\u2013water (1/2/5.5), Cl}2 (2), have been designed and both complexes show a slightly distorted octa\u00adhedral coordinated IrIII centre. The PCP pincer ligand system is arranged in a meridional manner, the CO ligand is located trans to the central PCP carbon and a hydride and chloride are located perpendicular above and below the P2C2 plane. With an Ir\u2014CCDP distance of 2.157\u2005(5)\u2005\u00c5, an Ir\u2014CO distance of 1.891\u2005(6)\u2005\u00c5 and a quite short C\u2014O distance of 1.117\u2005(7)\u2005\u00c5, complex 1 presents a strong carbonyl bond. Complex 2, the corresponding CH acid of 1, shows an additionally attached proton at the carbodi\u00adphospho\u00adrane carbon atom located anti\u00adperiplanar to the hydride of the metal centre. In comparison with complex 1, the Ir\u2014CCDP distance of 2.207\u2005(3)\u2005\u00c5 is lengthened and the Ir\u2014C\u2014O values indicate a weaker trans influence of the central carbodi\u00adphospho\u00adrane carbon atom.After the successful creation of the newly designed PCP carbodi\u00adphospho\u00adrane (CDP) ligand [Reitsamer al. 2012. Dalton al. 2007. Chem. C Consequently, this central atom disposes of two lone-electron pairs and is able to inter\u00adact with one or two Lewis acids functionality and can be described as a naked carbon atom or as a divalent carbon(0) atom in an excited singlet (12]Cl enters an oxidative addition reaction with Vaska\u2019s compound [IrI(CO)Cl(PPh3)2], forming the iridium PCP pincer CDP complex Cl (1) (see reaction scheme). During this reaction sequence, the central carbon atom is deprotonated, becomes neutral and coordin\u00adates the iridium transition metal. Treatment of complex 1 with hydro\u00adchloric acid leads to the protonation of the central carbon atom and consequently to the formation of the conjugate CH acid of 1, the Cl2 complex 2 1 (C(dppm)2-\u03ba3P,C,P\u2032)ClH]+ monocation and the chloride counter-ion. The iridium transition metal centre exhibits an octa\u00adhedral ligand system, formed by a meridional arranged C(dppm)2, relative to the C1 atom, a trans-coordinated carbonyl unit, and a chlorido and hydrido ligand located perpendicular to the meridional plane or more precisely trans to each other. The P1\u2014Ir1\u2014P4 angle of 170.69\u2005(5)\u00b0 indicates a small deviation from the octa\u00adhedral geometry and this value is larger compared to many related Iridium PCP pincer complexes. The environment of the CDP carbon atom C1 is strictly planar < Csp3]. Consequently, the Ir1\u2014C1 separation of 2.157\u2005(5)\u2005\u00c5 indicates an sp3 hybridization of the carbodiphosphorane carbon atom, which is substantiated by the data collected in Table\u00a01B] and 2.62\u2005\u00c5 [Cl2\u22efH3B] illustrate the location within the van der Waals radii. These C\u2014H\u22efX inter\u00adactions are a common feature of complexes containing dppm or related ligands ] and 2.47\u2005\u00c5 [Cl2\u22efH6B].Complex 1 Fig.\u00a01 crystall2 comprises two Cl2 complex mol\u00adecules (CH(dppm)2-\u03ba3P,C,P\u2032)ClH]2+ dication stabilized by two chloride counter-ions. Overall, complex 2 represents the conjugate CH acid of the Cl complex (1). The carbodi\u00adphospho\u00adrane carbon atom additionally coordinates a second Lewis acid, the proton H1, which adopts an anti-periplanar conformation relative to the hydrido ligand H11. As a consequence, atom C1 forms a distorted tetra\u00adhedron with the directly coordinated atoms (sum of angles = 344.3\u00b0). In comparison with complex 1, the values of the angles P2\u2014C1\u2013Ir1 and P3\u2014C1\u2014Ir1 are significantly reduced, whereas the P2\u2014C1\u2014P3 angles differs to a lesser extent (Table\u00a021 [2.157\u2005(5)\u2005\u00c5], as has also been observed in other carbodi\u00adphospho\u00adrane complexes ], 2.89\u2005\u00c5 [H2B\u22efCl7] and 2.57\u2005\u00c5 [H3B\u22efCl5].The asymmetric unit of es Fig.\u00a02, four mot Table\u00a02. The coos Table\u00a04 between 1, there are inter\u00adactions = 2.62\u2005\u00c5] with distances shorter than the sum of the van der Waals radii. Such C\u2014H\u22efX inter\u00adactions are a common feature of complexes containing dppm or related ligands = 2.59\u2005\u00c5 and H5B\u22efCl2 = 2.47\u2005\u00c5].In s Table\u00a03 between 2, C\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions ], 2.89\u2005\u00c5 [H2B\u22efCl7] and 2.57\u2005\u00c5 [H3B\u22efCl5]. A network of different inter\u00adactions occurs between the two independent complex mol\u00adecules. The water and hydro\u00adchloric acid solvent mol\u00adecules form hydrogen bonds with the chloride ligands or counter-ions and the hydrogen atoms of the complex mol\u00adecules, respectively.In s Table\u00a04 occur be2) using standard Schlenk techniques. The 1H, 13C and 31P NMR spectra were recorded on a Bruker DPX 300 NMR spectrometer and were referenced against the 13C/1H solvent peaks of the solvents chloro\u00adform, methanol or the external 85% H3PO4 standard, respectively. The phospho\u00adrus atoms in the NMR data are labelled as in Figs. 1All preparations were carried out under an inert atmosphere (N2-Synthesis of Cl (1): A mixture of 19.5\u2005mg of Vaska\u2019s complex (0.025\u2005mmol), 20.4\u2005mg of [CH(dppm)2]Cl (0.025\u2005mmol) was stirred at ambient temperature for 15\u2005min. The solvent was evaporated in vacuo and the residue was digested with a mixture of CH2Cl2 (0.1\u2005ml) and ethyl acetate (0.7\u2005ml). The solid was separated and washed twice with ethyl acetate (0.6\u2005ml). Single crystals were grown by slow evaporation of a solution in CH2Cl2. 31P {1H} NMR (CHCl3): \u03b4 31.9 , \u03b4 8.2 (P1/P4); 13C {1H} NMR (CDCl3): \u03b4 \u22124.4 = 4); 1H NMR (CDCl3): \u03b4 \u221216.7 .2-Synthesis of Cl2 (2): 19.5\u2005mg of Vaska\u2019s complex (0.025\u2005mmol) and 20.4\u2005mg of [CH(dppm)2]Cl (0.025\u2005mmol) . The mixture was stirred at ambient temperature for 15\u2005min. After addition of 0.1\u2005ml of hydro\u00adchloric acid (10\u2005mol\u2005L\u22121), the product crystallized upon standing for a day. 31P {1H} NMR (CHCl3/MeOH): \u03b4 45.3 , \u03b4 1.7 (P1/P4); 13C {1H} NMR (CDCl3): \u03b4 9.1 = 38, 1JC1H1 = 122); 1H NMR (CDCl3/MeOH): \u03b4 \u221218.9 .1 resulted in the location of the hydride hydrogen atom. The bond length was restrained to a distance of 1.6\u2005\u00c5 and a fixed isotropic displacement parameter of 1.5Ueq of iridium was applied. The hydrido ligand of complex 2 was also detected and refined isotropically without the use of bond restraints. Furthermore, the proton of the CDP carbon atom was spotted and refined with bond restraints of 0.98\u2005\u00c5. The hydrogen atoms of the water and solvent mol\u00adecules could only be partially detected and were omitted. A determination of a 1:1 positional disorder of one water mol\u00adecule (O4 and O4A) and one HCl or chloride (Cl10 and Cl1A) was possible. Eight chloride positions can be detected, which are occupied by a total of four chlorides and four hydro\u00adchloric acid units. The hydrogen-atom positions of the phenyl subunits and methyl\u00adene groups were refined with calculated positions (C\u2014H = 0.94 and 0.98\u2005\u00c5) using a riding model with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a052 are related to each other by the presence of a pseudo-symmetry centre. A halving of the c axis and consequently the changing of the monoclinic setting from P21/n to P21/c allows the consideration of one formula unit of 2. A closer observation of the sections of the reciprocal lattice along c* (l = 2n\u00a0+\u00a01) at different values of l results in the presence of frequent weak reflections. Consequently, an inter\u00adpretation of this system as three-dimensional network between two complex mol\u00adecules, four hydro\u00adchloric acid units and eleven water mol\u00adecules allows the involvement of these weak, but clearly existing reflections, and establishes the possibility of the distinction of the chloride and oxygen positions.The two complex mol\u00adecules of 10.1107/S2056989018004905/eb2007sup1.cifCrystal structure: contains datablock(s) global, complex1, complex2. DOI: 10.1107/S2056989018004905/eb2007complex1sup2.hklStructure factors: contains datablock(s) complex1. DOI: 10.1107/S2056989018004905/eb2007complex2sup3.hklStructure factors: contains datablock(s) complex2. DOI: 1577807, 1577808CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure is stabilized by N/C\u2014H\u22efO hydrogen bond, C\u2014H\u22efF and C\u2014H\u22ef\u03c0 inter\u00adactions with weak \u03c0\u2013\u03c0 inter\u00adactions contacts to form a three-dimensional architecture.The title compound, C 18H22FN5O8S, is used as a herbicide (pyrimidinyl\u00adsulfonyl\u00adurea herbicide). The dihedral angle between the mean planes of the pyridine and pyrimidine rings is 86.90\u2005(7)\u00b0. In the crystal, N/C\u2014H\u22efO hydrogen bonds, C\u2014H\u22efF and C\u2014H\u22ef\u03c0 inter\u00adactions link adjacent mol\u00adecules, forming a chain along [020]. A further two C\u2014H\u22efO hydrogen bonds together with weak \u03c0\u2013\u03c0 inter\u00adactions [ring centroid separation = 3.7584\u2005(12)\u2005\u00c5] further aggregate the structure into a three-dimensional architecture.The title compound, {systematic name: 1-[3-carbamo\u00adyl]amino}\u00adsulfon\u00adyl)pyridin-2-yl]-2-fluoro\u00adpropyl 2-meth\u00adoxy\u00adacetate}, C DOI: 10.1107/S2056989017012737/hg5495Isup2.hklStructure factors: contains datablock(s) I. DOI: 1572854CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound, a bromo derivative of pyran\u00adocoumarin, possesses photobiological activity. It was formed by bromination of seselin by using NBS in MeOH at room temperature. In the crystal, mol\u00adecules are linked by \u03c0\u2013\u03c0 stacking inter\u00adactions and weak C\u2014H\u22efO inter\u00adactions, forming layers parallel to (001). 15H14Br2O4 chromen-2(8H)-one], is a pyran\u00adocoumarin derivative formed by the bromination of seselin, which is a naturally occurring angular pyran\u00adocoumarin isolated from the Indian herb Trachyspermum stictocarpum. In the mol\u00adecule, the benzo\u00adpyran ring system is essentially planar, with a maximum deviation of 0.044\u2005(2)\u2005\u00c5 for the O atom. The di\u00adhydro\u00adpyran ring is in a half-chair conformation and the four essentially planar atoms of this ring form a dihedral angle of 4.6\u2005(2)\u00b0 with the benzo\u00adpyran ring system. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO hydrogen bonds, forming chains propagating along [010]. In addition, \u03c0\u2013\u03c0 stacking inter\u00adactions, with centroid\u2013centroid distances of 3.902\u2005(2) and 3.908\u2005(2)\u2005\u00c5, link the hydrogen-bonded chains into layers parallel to (001).The title compound, C The di\u00adhydro\u00adpyran ring C1\u2013C5/O1 is in a half-chair conformation and atoms C2 and C3 deviate by \u22120.385\u2005(4) and 0.280\u2005(4)\u2005\u00c5 from the plane through the other four essentially planar atoms (mean deviation 0.003\u2005\u00c5), which makes a dihedral angle of 4.6\u2005(2)\u00b0 with the benzo\u00adpyran ring system. The relative stereochemistry at atoms C3 and C4 is R/S and S/R.The title mol\u00adecule Fig.\u00a01 is compoCg1\u22efCg1 of 3.902\u2005(2)\u2005\u00c5 and Cg1\u22efCg2 of 3.908\u2005(2)\u2005\u00c5 where Cg1 and Cg2 are the centroids of the C1/C5/C6/C10\u2013C12 and O2/C6\u2013C10 rings, respectively, link the hydrogen-bonded chains, forming layers parallel (001) Fig.\u00a03.et al., 2016et al., 2006A search of the Cambridge Structural Database from ethyl acetate:hexane (1:4) at room temperature by slow evaporation of the solvents. NMR analysis: 1H NMR data : \u03b4H 8.02 , 7.32 , 6.82 , 5.36 , 4.26 , 3.56 , 1.50 , 1.54 .The title compound is a colourless solid substance formed on bromination of the naturally occurring seseline isolated from the methanol extract of iso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017006132/lh5842sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017006132/lh5842Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017006132/lh5842Isup3.cmlSupporting information file. DOI: 1545541CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular and crystal structure of zwitterionic 2-{[meth\u00adyl]carbamo\u00adyl}benzoate hemihydrate is reported. The crystal structure is stabilized by a variety of hydrogen bonds and offset \u03c0\u2013\u03c0 stacking inter\u00adactions. 15H15N3O3\u00b70.5H2O, comprises two 2-{[meth\u00adyl]carbamo\u00adyl}benzoate zwitterions (A and B) and a water mol\u00adecule. The dihedral angles between the pyridine and phenyl rings in the zwitterions are 53.69\u2005(10) and 73.56\u2005(11)\u00b0 in A and B, respectively. In the crystal, mol\u00adecules are linked by N\u2014H\u22efO, O\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22ef\u03c0(ring) hydrogen bonds into a three-dimensional network. The crystal structure also features \u03c0\u2013\u03c0 inter\u00adactions involving the centroids of the pyridine and phenyl rings [centroid\u2013centroid distances = 3.5618\u2005(12)\u2005\u00c5 in A and 3.8182\u2005(14)\u2005\u00c5 in B].The asymmetric unit of the title compound, C The study and synthesis of new zwitterions is therefore important in the search for new biomedical applications and a cocrystallized water mol\u00adecule, as shown in Fig.\u00a012\u2014Car single bond [1.38\u2005(3)\u2005\u00c5], but are similar to those found in related compounds with an N+=C double bond \u2005\u00c5 and C15A\u2014O2A = 1.257\u2005(3)\u2005\u00c5] in mol\u00adecule A, with comparable values in mol\u00adecule B, are similar to values found in other deprotonated carboxyl\u00adate groups meth\u00adyl]carbamo\u00adyl}benzoate zwitterions \u2005\u00c5, where Cg3 and Cg4 are the centroids of the N1A/C1A\u2013C5A and C9A\u2013C14A rings] Fig.\u00a04. The crys] Fig.\u00a05. An overet al., 2016N-[2-eth\u00adyl]phthalamic acid tetra\u00adhydrate phthalamic acid -2-[(1-phenyl\u00adeth\u00adyl)carbamo\u00adyl]benzoate -1-phenyl\u00adethanaminium 2-{carbamo\u00adyl}benzoate -4H-1\u03bb2-pyridine as the skeleton gave 15 hits, although none of these were zwitterionic derivatives comparable to the title compound. Of these, only three had aromatic rings in the cation in addition to the imino\u00adpoyridine unit phthalimide and 0.01\u2005mol of 4-amino-3-methyl\u00adpyridine in 10\u2005ml of di\u00admethyl\u00adformamide with a catalytic amount of potassium carbonate. The mixture was stirred in a 50\u2005ml round-bottomed flask at room temperature for about 3\u2005h. The progress of the reaction was monitored by thin-layer chromatography and the mixture was poured into cold water once the reaction was complete. The resulting precipitate was filtered off, washed successively with distilled water, and recrystallized from acetone solution by slow evaporation to obtain colourless block-shaped single crystals.The title compound was obtained unexpectedly from the reaction of 0.01\u2005mol of W were fixed at 0.86\u2005\u00c5, the H\u22efH distance was fixed at 1.34\u2005\u00c5 and the H atoms were refined with a riding model . The C-bound H atoms were positioned geometrically using a riding model, with Uiso(H) = 1.2 or 1.5Ueq(C) . A rotating-group model was applied to the methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017007836/sj5531sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989017007836/sj5531Isup2.hklStructure factors: contains datablock(s) I. DOI: 1552520CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "High resolution mass spectrometry (HRMS) was used to monitor library composition and establish conditions under which equilibria were established.Dynamic combinatorial chemistry (DCC) represents an approach, whereby traditional supramolecular scaffolds used for protein surface recognition might be exploited to achieve selective high affinity target recognition. Synthesis, in situ screening and amplification under selection pressure allows the generation of ligands, which bear different moieties capable of making multivalent non\u2010covalent interactions with target proteins. Generic tetracarboxyphenyl porphyrin scaffolds bearing four hydrazide moieties have been used to form dynamic combinatorial libraries (DCLs) using aniline\u2010catalyzed reversible hydrazone exchange reactions, in 10\u2009% DMSO, 5 m The use of different amino acids was intended to allow improved scaffold solubility and to investigate the role of functionality proximal to the hydrazone linker in terms of influencing exchange rate.A tetraphenylporphyrin scaffold was chosen as a multivalent hydrazone scaffold. This choice was motivated by the ready accessibility of commercial porphyrins with a sufficient number of points at which a hydrazide linker could be introduced, and the established utility of porphyrins in protein surface recognition.1\u20133aaaa were achieved by direct coupling of protected amino acid hydrazides with tetracarboxyphenylporphyrin, using PyBOP, to generate protected hydrazide porphyrins 4\u20136. Hydrazone scaffolds 1\u20133aaaa ready for DCC studies bearing a simple phenyl hydrazone at each exchangeable bond were obtained from the corresponding protected hydrazides 4\u20136 by TFA deprotection followed by reaction with benzaldehyde 7.Attempts to synthesise tetracarboxyhydrzidophenylporphyrin resulted in analytically intractable products (see Supporting information). Subsequent efforts centred on the introduction of an amino acid spacer between the porphyrin scaffold and the labile hydrazone linker. Initial synthesis using a glycine spacer resulted in formation of an insoluble product (see Supporting information). Therefore amino acid spacers containing groups to aid scaffold solubility, serine, aspartic acid and glutamic acid, were used. The introduction of the acid functionalities was also envisioned to aid the hydrazone exchange chemistry necessary during the DCL generation, through intramolecular proton transfer effects as has been observed in recent work from the Kool group.1\u20133aaaa, with three different aldehydes: 4\u2010carboxybenzaldehyde 8, 4\u2010methyl ester benzaldehyde 9 and 2,4\u2010dimethoxybenzaldehyde 10 was used to follow hydrazone exchange. Preliminary efforts to employ HPLC to monitor library distribution indicated that identification of gradients to separate closely related products would be challenging, and in the context of a screening workflow, HRMS represents a method to establish compound identity and qualitative changes in library composition in response to template. Hydrazone exchanges were performed with the three different porphyrin scaffolds 2aaaa with 8), hydrazone exchange was observed within 24 h, and could be visualised by HRMS to 8, at first increasing then decreasing. The products of successive hydrazone exchanges then followed in succession.Considering one of these systems, , with a shift to a greater number of hydrazone exchanges observed (see Supporting information).Having established effective exchange of the benzaldehyde moiety for another aldehyde it was necessary to assess the extent to which a thermodynamic rather than kinetic distribution of products was obtained. The speciation of porphyrins reacted with 25 equivalents of aldehyde for 24 h followed by addition of a further 25 equivalents of the same aldehyde and reaction for 24 h was monitored. Using 1\u20133aaaa were carried out using two aldehydes simultaneously to generate a DCL . Flash column chromatography was performed using silica gel 60 (0.043\u20130.063 mm VWR or Sigma Aldrich) or alumina (Brockman I from Sigma Aldrich), unless otherwise stated silica gel was used and pressure was applied by means of head bellows.Thin layer (silica) chromatography was performed using Merck Kiesegel 60 F1H NMR spectra were obtained on Bruker DPX 300 (300 MHz) Avance 500 (500 MHz) or DRX500 (500 MHz) spectrometers and referenced to either residual non\u2010deuterated solvent peaks or tetramethylsilane. 13C spectra were recorded on a Bruker DPX 300 (75 MHz) Bruker or an Avance 500 (126 MHz) and referenced to the solvent peak. 1H spectra are reported as follows: 1H NMR : \u03b4 = ppm to 2 d.p. . Chemical shifts are quoted in ppm with signal splitting recorded as singlet (s), doublet (d), triplet (t), quartet (q), multiplet (m), broad (br). Coupling constants, J, are measured to the nearest 0.1 Hz. Similarly, 13C spectra are reported as follows: \u03b4 : \u03b4 ppm to one decimal place. Infrared spectra were recorded on a Perkin\u2013Elmer Fourier\u2010Transfer spectrometer. Spectra were analysed neat and structurally important absorptions are quoted. Absorption maxima (\u03bd\u0303max=) are quoted in wavenumbers (cm \u20131). HPLC LC\u2010MS were recorded on a Bruker HCT ultra under electrospray ionisation (ESI) conditions. High resolution mass spectra were recorded on a Bruker Daltonics micrOTOF Premier Mass Spectrometer, under positive ESI conditions unless otherwise stated.tert\u2010butoxy)\u20101\u2010{N\u2032\u2010[(tert\u2010butoxy)carbonyl]hydrazinecarbonyl}ethyl)carbamoyl]phenyl}\u201017\u2010{4\u2010[((1S)\u20102\u2010(tert\u2010butoxy)\u20101\u2010{N\u2032\u2010[(tert\u2010butoxy)carbonyl]hydrazinecarbonyl}ethyl)carbamoyl]phenyl}\u201021,22,23,24\u2010tetraazapentacyclotetracosa\u20101,3(24),4,6,8,10,12,14,16(22),17,19\u2010undecaen\u20102\u2010yl)\u2010N\u2010((1S)\u20102\u2010(tert\u2010butoxy)\u20101\u2010{N\u2032\u2010[(tert\u2010butoxy)carbonyl]hydrazinecarbonyl}ethyl)benzamide (4):4\u2010\u20102\u2010, PyBOP , tert\u2010butyl 2\u2010(2\u2010amino\u20103\u2010tert\u2010butoxypropanoyl)hydrazinecarboxylate, and diisopropylethylamine in anhydrous dimethylformamide (5 mL) were stirred under a nitrogen atmosphere for 18 h. The reaction mixture was then diluted with ethyl acetate (50 mL) and washed with saturated sodium hydrogen carbonate solution (50 mL), 1 m hydrochloric acid (50 mL) and brine (3\u2009\u00d7\u200950 mL). The organic phase was dried (sodium sulphate) and concentrated to yield the crude product as a purple solid. This was purified by flash column chromatography (1:1 ethyl acetate/dichloromethane then ethyl acetate) to yield the product as a purple solid . 1H NMR : \u03b4 = 1.29 , 1.45 , 3.51\u20133.63 , 4.07 , 4.83 , 6.40\u20136.57 , 8.15 , 8.21 , 8.40 , 8.73 ppm. 13C NMR : \u03b4 = 26.8, 27.5, 38.4, 53.3, 61.6, 74.0, 80.8, 106.6, 119.3, 125.8, 133.4, 134.4, 145.2, 156.2, 168.4, 170.8 ppm. IR (solid state): \u03bd\u0303 = 3251 (N\u2013H), 1696 (C=O carbamate), 1645 (C=O amide) cm\u20131. ESI\u2010MS m/z found 1820.9402 [M + 2H]+, [C96H124N16O20]+ calcd. 1820.9178.N\u2032\u2010[(1E)\u2010phenylmethylidene]hydrazinecarbonyl}ethyl)carbamoyl]phenyl}\u201017\u2010{4\u2010[((1S)\u20102\u2010hydroxy\u20101\u2010{N\u2032\u2010[(1E)\u2010phenylmethylidene]hydrazinecarbonyl}ethyl)carbamoyl]phenyl}\u201021,22,23,24\u2010tetraazapentacyclotetracosa\u20101,3(24),4,6,8,10,12,14,16(22),17,19\u2010undecaen\u20102\u2010yl)\u2010N\u2010((1S)\u20102\u2010hydroxy\u20101\u2010{N\u2032\u2010[(1E)\u2010phenylmethylidene]hydrazinecarbonyl}ethyl)benzamide (1aaaa):4\u2010\u20102\u2010hydroxy\u20101\u2010{4 in trifluoroacetic acid (4.5 mL), water (0.25 mL) and triisopropylsilane (0.25 mL) was stirred for 6 h. The solution was then concentrated to yield the deprotected hydrazide as a green solid (complete deprotection was confirmed by HRMS). The green solid was then redissolved in water (2 mL) and acetonitrile (2 mL) and benzaldehyde (2 drops) was added, the mixture was then stirred for 30 min. The precipitate was isolated and washed with water and acetonitrile to yield the product as a dark purple solid . 1H NMR : \u03b4 = 1.67 , 3.94 , 4.76 , 5.63 , 7.51 , 7.63 , 7.75 , 7.95 , 8.12 , 8.39 , 8.84 , 8.91 , 11.56 , 11.73 (cis and trans hydrazone isomers observed in 1:1 ratio). IR (solid state): \u03bd\u0303 = 3212 (N\u2013H), 1633 (C=O amide), 1608 (C=N) cm\u20131. ESI\u2010MS m/z found 1547.5750 [M + H]+, [C88H75N16O12]+: calcd. 1547.5340.tert\u2010Butyl (3S)\u20103\u2010{[4\u2010\u20103\u2010(tert\u2010butoxy)\u20101\u2010{N\u2032\u2010[(tert\u2010butoxy)carbonyl]hydrazinecarbonyl}\u20103\u2010oxopropyl)carbamoyl]phenyl}\u201017\u2010{4\u2010[((1S)\u20103\u2010(tert\u2010butoxy)\u20101\u2010{N\u2032\u2010[(tert\u2010butoxy)carbonyl]hydrazinecarbonyl}\u20103\u2010oxopropyl)carbamoyl]phenyl}\u201021,22,23,24\u2010tetraazapentacyclotetracosa\u20101,3(24),4,6,8,10,12,14,16(22),17,19\u2010undecaen\u20102\u2010yl)phenyl]formamido}\u20103\u2010{N\u2032\u2010[(tert\u2010butoxy)carbonyl]hydrazinecarbonyl}propanoate (5): Tetracarboxyphenylporphyrin , PyBOP , (S)\u2010tert\u2010butyl 2\u2010(2\u2010amino\u20104\u2010tert\u2010butoxy\u20104\u2010oxobutanoyl)hydrazinecarboxylate, and anhydrous diisopropylethylamine in anhydrous dimethylformamide (5 mL) were stirred under a nitrogen atmosphere for 18 h. The reaction mixture was then diluted with ethyl acetate (50 mL) and washed with saturated sodium hydrogen carbonate solution (50 mL), 1 m hydrochloric acid (50 mL) and brine (3\u2009\u00d7\u200950 mL). The organic phase was dried (sodium sulphate) and concentrated to yield the crude product as a purple solid. This was purified by flash column chromatography (1:1 ethyl acetate/dichloromethane then ethyl acetate) to yield the product as a purple solid . 1H NMR : \u03b4 = 1.53 , 1.55 , 2.95 , 3.10 , 5.30 , 5.45 , 8.21 , 8.27 , 8.73 , 9.02 . 13C NMR : \u03b4 = 27.4, 27.6, 37.2, 49.3, 80.9, 81.5, 119.3, 125.9, 133.2, 134.4, 145.4, 149.5, 156.3, 168.4, 170.2, 171.3, 185.1 ppm. IR (solid state): \u03bd\u0303 = 3275 (N\u2013H), 1723 (C=O ester), 1711 (C=O carbamate), 1647 (C=O amide) cm\u20131. ESI\u2010MS m/z found 1931.9891 [M + H]+, [C100H123N16O24]+: calcd. 1931.8896.N\u2032\u2010[(1E)\u2010phenylmethylidene]hydrazinecarbonyl}ethyl)carbamoyl]phenyl}\u201017\u2010{4\u2010[((1S)\u20102\u2010carboxy\u20101\u2010{N\u2032\u2010[(1E)\u2010phenylmethylidene]hydrazinecarbonyl}ethyl)carbamoyl]phenyl}\u201021,22,23,24\u2010tetraazapentacyclotetracosa\u20101,3(24),4,6,8,10,12,14,16(22),17,19\u2010undecaen\u20102\u2010yl)phenyl]formamido}\u20133\u2010{N\u2032\u2010[(1E)\u2010phenylmethylidene]hydrazinecarbonyl}propanoic Acid (2aaaa):3(S)\u20103\u2010{+, [C92H75N16O16]+: calcd. 1659.5547.tert\u2010Butyl (4S)\u20104\u2010{[4\u2010\u20101\u2010{N\u2032\u2010[(tert\u2010butoxy)carbonyl]hydrazinecarbonyl}\u20104\u2010oxobutyl)carbamoyl]phenyl}\u201017\u2010{4\u2010[((1S)\u20104\u2010(tert\u2010butoxy)\u20101\u2010{N\u2032\u2010[(tert\u2010butoxy)carbonyl]hydrazinecarbonyl}\u20104\u2010oxobutyl)carbamoyl]phenyl}\u201021,22,23,24\u2010tetraazapentacyclotetracosa\u20101,3(24),4,6,8,10,12,14,16(22),17,19\u2010undecaen\u20102\u2010yl)phenyl]formamido}\u20104\u2010{N\u2032\u2010[(tert\u2010butoxy)carbonyl]hydrazinecarbonyl}butanoate (6): {4\u2010+, [C104H131N16O24]+: calcd. 1987.9522.N\u2032\u2010[(1E)\u2010phenylmethylidenehydrazinecarbonyl]propyl}carbamoyl)phenyl]\u201017\u2010{4\u2010[((1S)\u20103\u2010carboxy\u20101\u2010{N\u2032\u2010[(1E)\u2010phenylmethylidene]hydrazinecarbonyl}propyl)carbamoyl]phenyl}\u201021,22,23,24\u2010tetraazapentacyclotetracosa\u20101,3(24),4,6,8,10,12,14,16(22),17,19\u2010undecaen\u20102\u2010yl)phenyl]formamido}\u20104\u2010{N\u2032\u2010[(1E)\u2010phenylmethylidene]hydrazinecarbonyl}butanoic Acid (3aaaa):(4S)\u20104\u2010{4\u2010+, [C96H83N16O16]+: calcd. 1715.6173.Hydrazone Exchange Studies: Hydrazone exchange reactions were carried out in HPLC vials and were followed using high resolution mass spectrometry on a Bruker Daltonics micrOTOF Premier Mass Spectrometer, using 10 \u00b5L injections and summing the masses over a range of 1.0 to 3.0 min. The intensity of the maximum peak for each of the successive hydrazone exchanges was taken and the percentage associated with each of the porphyrin peaks calculated.1\u20133aaaa were made up to 5 mm concentration in DMSO and were stored in plastic Eppendorf tubes. 1 m stocks of catalyst (11 and 12) and aldehydes were made up in DMSO.Hydrazone functionalised porphyrins 1\u20133aaaa in DMSO was added to a solution of the aldehyde and catalyst to give a total concentration of 10\u2009% DMSO in 5 mm ammonium acetate buffer, pH 6.75, to a final porphyrin concentration of 100 \u00b5m and stated concentrations of other components. For time courses, mass spectra were obtained at appropriates time points . For measurements at single time points, mass spectra were obtained after 24 h incubation. For pre\u2010incubated samples, the pre\u2010incubated mixture was left for 24 h, and a mass spectrum obtained, prior to addition of further components.In all exchange reactions the hydrazone functionalised porphyrin Supporting InformationClick here for additional data file."} +{"text": "The asymmetric unit of the title compound consists of one and a half bis\u00adchalcone mol\u00adecules. In the crystal, mol\u00adecules are linked into a three-dimensional network by C\u2014H\u22efF and C\u2014H\u22efO hydrogen bonds, some of the C\u2014H\u22efF links being unusually short (< 2.20\u2005\u00c5). Hirshfeld surface analyses are presented and discussed. 24H14F4O2, comprises of one and a half mol\u00adecules; the half-mol\u00adecule is completed by crystallographic inversion symmetry. In the crystal, mol\u00adecules are linked into a three-dimensional network by C\u2014H\u22efF and C\u2014H\u22efO hydrogen bonds. Some of the C\u2014H\u22efF links are unusually short (< 2.20\u2005\u00c5). Hirshfeld surface analyses for the title compound are presented and discussed.The asymmetric unit of the title compound, C They consist of two aromatic rings joined by a three-carbon-atom unsat\u00adurated carbonyl system (\u2013CH=CH\u2014CO\u2013). Bischalcones with the general formula Ar\u2014CH=CH\u2014CO\u2014CH=CH\u2014Ar A, the carbonyl groups form similar torsion angles with the 2,4-di\u00adfluoro\u00adphenyl ring and the olefinic double bond . Conversely, the torsion angles between the olefinic double bond and the central benzene ring are slightly different . This leads to slight differences in the dihedral angles between the terminal 2,4-di\u00adfluoro\u00adphenyl and the central benzene rings [7.91\u2005(2)\u00b0 for C1A\u2013C6A and 6.28\u2005(2)\u00b0 for C19A\u2013C24A]. In mol\u00adecule B, both torsion angles \u03c41 and \u03c43 are comparable to those in mol\u00adecule A . However, mol\u00adecule B is slightly closer to planar than mol\u00adecule A, as its central and terminal rings subtend a dihedral angle of 5.49\u2005(2)\u00b0. This might arise from the lower torsion angle between the olefinic double bond and the central benzene ring [O1B\u2014C7B\u2014C8B\u2014C9B = 0.9\u2005(6)\u00b0]. Selected torsion and dihedral angles are listed in Table\u00a01et al., 2005The asymmetric unit of (I)B) Fig.\u00a01. The molA\u2014H8A\u22efF1A, C17A\u2014H17A\u22efF3A and C8B\u2014H8B\u22efF1B hydrogen bonds generates an S(6) ring motif s Table\u00a01 generates Table\u00a02. As the er Fig.\u00a02. Atom F2et al., 2004CrystalExplorer 3.1 and outside (de) of the Hirshfeld surface analyze the inter\u00admolecular inter\u00adaction via the mapping of dnorm. In a dnorm surface, any inter\u00admolecular inter\u00adactions will appear as a red spot.The Hirshfeld surface analyses . Similarly, the C\u2014H\u22efF inter\u00adactions are identified by red spots near the F2A atom in mol\u00adecule A . As illustrated in Fig.\u00a05de and di diagonal axes represent the overall two-dimensional FP and those delineated into F\u22efH/H\u22efF, H\u22efH and O\u22efH/H\u22efO contacts, respectively. The most significant inter\u00admolecular inter\u00adactions are the reciprocal F\u22efH/H\u22efF inter\u00adactions (30.1%), which appear as two sharp symmetric spikes in FP maps with a prominent long spike at de + di \u2243 2.3\u2005\u00c5 . The H\u22efH inter\u00adactions appear in the central region of the FP with ed = id \u2243 2.4\u2005\u00c5 and contribute 29.0% to the Hirshfeld surface whereas two symmetrical narrow pointed wings corresponding to the O\u22efH/H\u22efO inter\u00adactions with 12.7% contribution appear at diagonal axes of de + di \u2243 2.4\u2005\u00c5 . The percentage contributions for other inter\u00admolecular contacts are less than 10% in the Hirshfeld surface mapping .Dark-red spots that are close to atoms O1ds Fig.\u00a04a. Simil A Fig.\u00a04b. As il\u2005\u00c5 Fig.\u00a05b. The Hce Fig.\u00a05c wherea\u2005\u00c5 Fig.\u00a05d. The pet al., 2016E)-1-(4-fluoro\u00adphen\u00adyl)-3-phenyl\u00adprop-2-en-1-one as the main skeleton revealed the presence of seven structures containing the chalcone moiety with different substituent similar to the title compounds in this study. These structures are 4\u2032-fluoro\u00adchalcone -3-[4-(di\u00admethyl\u00adamino)\u00adphen\u00adyl]-1-(4fluoro\u00adphen\u00adyl)prop-2-en-1-one -3-(4-chloro\u00adphen\u00adyl)-1-(4-fluoro\u00adphen\u00adyl)prop-2-en-1-one phen\u00adyl]prop-2-en-1-ones -1-(4-fluoro\u00adphen\u00adyl)-3-(4-methyl\u00adphen\u00adyl)prop-2-en-1-one in methanol (20\u2005ml) was mixed with 2,4-di\u00adfluoro\u00adaceto\u00adphenone (0.02\u2005mol) in methanol (20\u2005ml) in the presence of NaOH. The reaction mixtures were stirred for about 5\u20136\u2005h at room temperature. The resultant crude products were filtered, washed successively with distilled water and recrystallized from ethanol solution to get the title compound. Yellow blocks of (I)-3,3\u2032-bis\u00adprop-2-en-1-one), C Solvent for growing crystals: mixture of chloro\u00adform and aceto\u00adnitrile (1:1 v/v); yield 85%, m.p. 447\u2013449\u2005K; FT\u2013IR (ATR (solid) cm\u22121): 3101 , 1600 , 1593, 1420 , 1229 ; 1H NMR : \u03b4 7.969\u20137.922 , 7.818\u20137.787 , 7.697 , 7.059\u20137.022 , 6.969\u20136.935 ; 13C NMR : 187.00 (C7), 143.62 (C9), 136.83 (C2), 133.11 (C10), 133.03 (C5), 129.14 , 126.18 (C6), 126.12 (C8) 112.47, 112.27 (C3), 105.01, 104.81 (C1), 104.59 (C4).Uiso(H) = 1.2Ueq(C) for H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901701564X/hb7696sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901701564X/hb7696Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901701564X/hb7696Isup3.cmlSupporting information file. DOI: 1449628CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound, tetra\u00adhydro\u00adseselin (THS), a hydrogenated product of the angular pyran\u00adocoumarin seselin, possesses photo-biological activity against different kinds of inflammatory skin diseases such as atopic dermatitis and pigment disorders like vitiligo and psoriasis. 14H16O3, a pyran\u00adocoumarin chromen-2-one] obtained from the hydrogenation of seselin in the presence of Pd/C in MeOH at room temperature, the dihedral angle between the central benzene ring and the best planes of the outer fused ring systems are 6.20\u2005(7) and 10.02\u2005(8)\u00b0. In the crystal, mol\u00adecules show only very weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions.In the title compound, tetra\u00adhydro\u00adseselin, C HL60 and A431, has been observed cyclo\u00adaddition] and diadducts [(4\u00a0+\u00a02) cyclo\u00adaddition] and 178.73\u2005(14)\u00b0, respectively, indicating that these rings are coplanar. The destruction of photo-biological activity and change of conformation of the pyran rings of the title mol\u00adecule is considered to be due to the loss of the double bonds in seselin.In the title compound, the three different fused rings comprising the mol\u00adecule Fig.\u00a01, are thei (ring) [3.221\u2005(2)\u2005\u00c5] and methyl\u00adene C9\u2014H\u22efO3i (carbon\u00adyl) [3.412\u2005(2)\u2005\u00c5] inter\u00adactions and methyl\u00adene C8\u2014H\u22efO3ii (carbon\u00adyl) [3.593\u2005(3)\u2005\u00c5] inter\u00adactions -[1]-benzo\u00adpyran-7-one; bromo\u00adhydroxy-seselin = 1.2Ueq(C). Those on methyl groups were allowed to ride with C\u2014H = 0.96\u2005\u00c5 and with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S205698901700932X/zs2379sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901700932X/zs2379Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901700932X/zs2379Isup3.cmlSupporting information file. DOI: 1557474CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N\u2014H\u22efO, O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds play an important role in the crystal packing, resulting in the formation of mol\u00adecular sheets parallel to the bc plane.The mol\u00adecules of C 15H14N2O3, crystallizes with two independent mol\u00adecules (A and B) in the asymmetric unit that differ in the orientation of the 3-meth\u00adoxy\u00adphenyl group with respect to the methyl\u00adidenebenzohydrazide unit. The dihedral angles between the two benzene rings are 24.02\u2005(10) and 29.30\u2005(9)\u00b0 in mol\u00adecules A and B, respectively. In mol\u00adecule A, the meth\u00adoxy group is twisted slightly relative to its bound benzene ring, with a Cmeth\u00adyl\u2014O\u2014C\u2014C torsion angle of 14.2\u2005(3)\u00b0, whereas it is almost co-planar in mol\u00adecule B, where the corresponding angle is \u22122.4\u2005(3)\u00b0. In the crystal, the mol\u00adecules are linked by N\u2014H\u22efO, O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds, as well as by weak C\u2014H\u22efO inter\u00adactions, forming sheets parallel to the bc plane. The N\u2014H\u22efO hydrogen bond and weak C\u2014H\u22efO inter\u00adaction link different mol\u00adecules (A\u22efB) whereas both O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds link like mol\u00adecules (A\u22efA) and (B\u22efB). Pairs of inversion-related B mol\u00adecules are stacked approximately along the a axis by \u03c0\u2013\u03c0 inter\u00adactions in which the distance between the centroids of the 3-meth\u00adoxy\u00adphenyl rings is 3.5388\u2005(12)\u2005\u00c5. The B mol\u00adecules also participate in weak C\u2014H\u22ef\u03c0 inter\u00adactions between the 4-hy\u00addroxy\u00adphenyl and the 3-meth\u00adoxy\u00adphenyl rings.The title compound, C Various derivatives of benzohydrazide have been reported to possess a range of biological properties, including anti\u00adbacterial -4-hy\u00addroxy-N\u2032-(3-hy\u00addroxy-4-meth\u00adoxy\u00adbenzyl\u00adidene)benzohydrazide -4-hy\u00addroxy-N\u2032-benzohydrazide A and B, of the title benzohydrazide derivative, C15H14N2O3, in the asymmetric unit of (I)A and B, respectively. The mol\u00adecules exist in the trans-conformation with respect to the C8=N2 bond [1.275\u2005(2)\u2005\u00c5 in mol\u00adecule A and 1.271\u2005(2)\u2005\u00c5 in mol\u00adecule B] and the torsion angle N1\u2014N2\u2014C8\u2014C9 = \u2212178.14\u2005(16)\u00b0 in mol\u00adecule A and \u2212177.69\u2005(16)\u00b0 in mol\u00adecule B. Five atoms of the central fragment are approximately coplanar, having r.m.s. deviations of 0.0179\u2005(2)\u2005\u00c5 in mol\u00adecule A and 0.0327\u2005(2)\u2005\u00c5 in mol\u00adecule B. The mean plane through this central fragment makes dihedral angles of 23.87\u2005(11) and 0.20\u2005(12)\u00b0 with the planes of the 4-hy\u00addroxy\u00adphenyl and 3-meth\u00adoxy\u00adphenyl rings, respectively, in mol\u00adecule A. The corresponding values are 22.58\u2005(11) and 11.04\u2005(11) \u00b0 in mol\u00adecule B. In mol\u00adecule A, the meth\u00adoxy group is slightly twisted from the attached benzene ring [C15\u2014O3\u2014C11\u2014C10 = 14.2\u2005(3)\u00b0] whereas it is essentially coplanar in mol\u00adecule B [where the corresponding torsion angle is \u22122.4\u2005(3)\u00b0]. The bond distances agree with literature values and are comparable with those in related structures whereas the O2A\u2014H2A\u22efN2Aiii and O2A\u2014H2A\u22efO1Aiii hydrogen bonds link equivalent A mol\u00adecules, and O2B\u2014H2B\u22efN2Bii and O2B\u2014H2B\u22efO1Bii hydrogen bonds link equivalent B mol\u00adecules. Stacking of planes of mol\u00adecules in the a-axis direction involves \u03c0\u2013\u03c0 inter\u00adactions between B mol\u00adecules with Cg\u22efCgvi distance of 3.5388\u2005(12)\u2005\u00c5. A weak C\u2014H\u22ef\u03c0 inter\u00adaction (C3B\u2014H3B\u22efCgv) between the 4-hy\u00addroxy\u00adphenyl ring and the 3-meth\u00adoxy\u00adphenyl ring of symmetry-related B mol\u00adecules is also present x, y, \u2212z; (iii) x, y, \u2212z; (iv) x, 1\u00a0+\u00a0y, z; (v) \u2212x, \u2212y, z; (vi) 1\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y, 2\u00a0\u2212\u00a0z; Cg is the centroid of the C9B\u2013C14B ring].In the crystal Fig.\u00a02, the molnt Fig.\u00a03 [symmetret al., 2011et al., 2011et al., 2012Chemical context section.A search of SciFinder in ethanol (10\u2005ml) and 3-meth\u00adoxy\u00adbenzaldehyde in ethanol (10\u2005ml) were mixed, stirred and refluxed for 5\u2005h. The resulting mixture was then cooled to room temperature. The white precipitate that formed was filtered. Colorless block-shaped single crystals of (I)3OH) \u03bbmax (log\u220a): 212 (5.51), 302 (5.61)\u2005nm; FT\u2013IR (KBr) \u03bd: 3158, 2834, 1648, 1607, 1509\u2005cm\u22121; 1H NMR \u03b4: 11.65 , 10.15 , 8.39 , 7.80 , 7.27 , 7.25 , 7.37 , 7.00 , 6.86 , 3.81 p.p.m.UV\u2013Vis N\u2032-benzyl\u00adidenebenzohydrazide skeleton by showing the characteristic signals of an amine (N=CH) at 8.39 and an amide (N\u2014H) at 11.65 p.p.m.The UV\u2013Vis spectrum of (I)et al. = 0.85 or 0.87\u2005\u00c5; d(O\u2014H) = 0.82\u2005\u00c5; d(C\u2014H) = 0.93\u2005\u00c5 for aromatic and CH; and 0.96\u2005\u00c5 for CH3 atoms. The Uiso values were constrained to be 1.5Ueq of the carrier atom for methyl and hydroxyl H atoms, and 1.2Ueq for the remaining H atoms. A rotating group model was used for the methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016013268/pk2585sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989016013268/pk2585Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016013268/pk2585Isup3.cmlSupporting information file. DOI: 1499671CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E conformation about the C=C bond, and the quinoline ring system and the benzene ring are inclined to one another by 29.22\u2005(7)\u00b0.The title 8-hy\u00addroxy\u00adquinoline derivative has an 19H15NO3, was synthesized by a Perkin reaction of 2-methyl-8-hy\u00addroxy\u00adquinoline and 4-formyl-2-methyl\u00adbenzoate in acetic anhydride under a nitro\u00adgen atmosphere. The mol\u00adecule has an E conformation about the C=C bond, and the quinoline ring system and the benzene ring are inclined to one another by 29.22\u2005(7)\u00b0. There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond in the 8-hy\u00addroxy\u00adquinoline moiety. In the crystal, mol\u00adecules are linked by pairs of O\u2014H\u22efO hydrogen bonds, forming inversion dimers with an R22(28) ring motif. The dimers are linked by C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming sheets parallel to plane (10-1).The title compound, C The C11\u2014C10 and C6\u2014C9 bond lengths are 1.463\u2005(2) and 1.466\u2005(2)\u2005\u00c5, respectively. These distances are shorter than the standard length of a C\u2014C single bond (ca 1.5\u2005\u00c5) because of the conjugate system formed by the C9=C10 bond and the aromatic systems. The quinoline ring system and the benzene ring are inclined to one another by 29.22\u2005(7)\u00b0.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01In the crystal, mol\u00adecules are linked by pairs of O\u2014H\u22efO hydrogen bonds, forming inversion dimers with an f Table\u00a01. The dimet al., 2016E conformation about the C=C bond, and in the crystal they also form inversion dimers. They include 2-{2-[4-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl]vin\u00adyl}quinolin-8-ol = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901601210X/su5312sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901601210X/su5312Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901601210X/su5312Isup3.molSupporting information file. DOI: Click here for additional data file.10.1107/S205698901601210X/su5312Isup4.cmlSupporting information file. DOI: 859030CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I atom in the title methanol solvate is coordinated octa\u00adhedrally by two N atoms of the chelating organic ligand, one Br atom and three facially configured carbonyl ligands. Hydrogen bonds between the complex and methanol solvent mol\u00adecules lead to a layered arrangement in the structure.The Re 16H16N4O3)(CO)3]\u00b7CH3OH, the ReI atom adopts a distorted octa\u00adhedral coordination sphere with a facial arrangement of the three carbonyl ligands. Two N atoms of the chelating 5--3-(pyridin-2-yl)-1H-1,2,4-triazole ligand and two carbonyl ligands define the equatorial plane of the complex, with the third carbonyl ligand and the bromide ligand in axial positions. Conventional hydrogen bonds including the methanol solvent mol\u00adecules assemble the complex mol\u00adecules through mutual N\u2014H\u22efO\u2014H\u22efBr links [N\u22efO = 2.703\u2005(3)\u2005\u00c5 and O\u22efBr = 3.255\u2005(2)\u2005\u00c5] into centrosymmetric dimers, whereas weaker C\u2014H\u22efO and C\u2014H\u22efBr hydrogen bonds [C\u22efO = 3.215\u2005(3)\u20133.390\u2005(4)\u2005\u00c5 and C\u22efBr = 3.927\u2005(3)\u2005\u00c5] connect the dimers into double layers parallel to the (111) plane.In the title compound, [ReBr(C The two N atoms and two carbonyl C atoms define the equatorial plane, while the octa\u00adhedral coordination sphere is completed by the third carbonyl C atom and the Br atom [Re\u2014Br = 2.6222\u2005(3)\u2005\u00c5] in axial positions. The CO ligands are almost linearly coordinated, with O\u2014C\u2014Re bond angles of 178.2\u2005(3), 177.8\u2005(3) and 177.8\u2005(3)\u00b0. The C\u2014Re\u2014C bond angles between carbonyl C atoms are 90.6\u2005(1), 90.2\u2005(1) and 88.7\u2005(1)\u00b0, close to ideal values, whereas the cis equatorial bite angle of the chelating ligand (N1\u2014Re1\u2014N4) is 73.42\u2005(8)\u00b0.The three carbonyl ligands bonded to the ReL)(CO)3] . However, this situation is unlikely to be a consequence of slipped \u03c0\u2013\u03c0 inter\u00adactions, since the corresponding slippage angle exceeds 56\u00b0 and the inter\u00adcentroid distance is as long as Cg(C6\u2013C10/N4)\u22efCg(C4/C5/N1\u2013N3)v = 4.090\u2005(3)\u2005\u00c5 . Although such weak inter\u00adactions are characteristic of related metal\u2013carbonyl structures in benzene (30\u2005ml). The mixture was refluxed for 4\u2005h under a stream of argon and then allowed to cool to room temperature. The yellow product was collected by suction filtration, washed with hexane and dried . Crystals suitable for X-ray diffraction were obtained by slow diffusion of hexane into a methanol\u2013di\u00adchloro\u00admethane solution of the complex. IR : \u03bdas(CO) 2028 (s), \u03bds(CO) 1894 (s). 1H NMR : \u03b4 9.02 , 8.39 , 8.35 , 7.75 , 7.44 , 3.93 .Penta\u00adcarbonyl\u00adrhenium(I) bromide was mixed with 5--3-(pyridin-2-yl)-1Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq otherwise. The O-bound H atom of the methanol solvent mol\u00adecule was found from a difference map and was refined with O\u2014H = 0.95\u2005\u00c5 and Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017003371/wm5372sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017003371/wm5372Isup2.hklStructure factors: contains datablock(s) I. DOI: 1535292CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Graphene plasmonics has become a highlighted research area due to the outstanding properties of deep-subwavelength plasmon excitation, long relaxation time, and electro-optical tunability. Although the giant conductivity of a graphene layer enables the low-dimensional confinement of light, the atomic scale of the layer thickness is severely mismatched with optical mode sizes, which impedes the efficient tuning of graphene plasmon modes from the degraded light-graphene overlap. Inspired by gap plasmon modes in noble metals, here we propose low-dimensional hybrid graphene gap plasmon waves for large light-graphene overlap factor. We show that gap plasmon waves exhibit improved in-plane and out-of-plane field concentrations on graphene compared to those of edge or wire-like graphene plasmons. By adjusting the chemical property of the graphene layer, efficient and linear modulation of hybrid graphene gap plasmon modes is also achieved. Our results provide potential opportunities to low-dimensional graphene plasmonic devices with strong tunability. In the context of light-matter interactions, the concentration of electromagnetic fields on materials is a critical issue for the performance of tunable optical devices, such as photodetectors3678Due to its two-dimensional (2D) structure with extremely large conductivity from the massless Dirac point111314161719212223242126272931Here, we focus on low-dimensional waveguide systems for the improved light-graphene overlap factor. We firstly reveal the existence of hybrid graphene gap plasmon (H-GGP) modes the field profile of which is strongly confined inside the graphene gap between metallic and dielectric graphene layers. We demonstrate that the H-GGP mode has larger field concentration on graphene layers than those of edge33w (the sheet conductivity \u03c3(G) where Im{\u03c3(G)}\u2009<\u20090) is inserted in-between semi-infinite metallic domain M (Im{\u03c3(M)}\u2009>\u20090) and dielectric domain D (Im{\u03c3(D)}\u2009<\u20090). The gap region G satisfies the condition of Im{\u03c3(D)}\u2009<\u2009Im{\u03c3(G)}\u2009<\u20090, as the analogy of plasmonic gap modes in noble metals8212627\u03bc(x). \u03bc-dependency of the graphene conductivity calculated by Kubo formula1121f\u2009=\u2009\u03c9/2\u03c0\u2009=\u200920\u2009THz, charged particle scattering rate8\u03a9(M))\u22121\u2009>\u20090.6 and 0.5\u2009<\u2009(\u03a9(D))\u22121\u2009<\u2009(\u03a9(G))\u22121\u2009<\u20090.6, where \u03a9\u22121\u2009=\u2009\u03bc/(\u210f\u03c9) is the normalized chemical potential.We consider the 2D metal-gap-dielectric waveguide system composed of three distinct graphene domains ; a dielew\u2009=\u20095\u2009nm, (\u03a9(M))\u22121\u2009=\u20094, (\u03a9(G))\u22121\u2009=\u20090.54, and (\u03a9(D))\u22121\u2009=\u20090.5002). In the numerical analysis, the graphene is considered as the film\u03b4\u2009=\u20090.2\u2009nm and the relative bulk permittivity of \u03b5g(\u03c9)\u2009=\u20091\u2009+\u2009j\u03c3g(\u03c9)/(\u03c9\u03b50\u03b4), where \u03c3g is sheet conductivity of graphene obtained from Kubo formula33\u03c3~\u2009\u2212\u2009\u03c3 for all z) have improved confinement compared to that of the GEP mode with much stronger structural asymmetry |\u2009\u226a\u2009|\u03c3| and |\u03c3|\u2009=\u2009|\u03c3| for z\u2009\u2260\u20090), H-GGP exhibits more confined transverse (Ex) field on the gap region than that of the 1D-SPP mode, as similar to the difference between gap plasmons and surface plasmons in noble metals8Im{\u03c3(G)}\u00b7Ex(G)~Im{\u03c3(D)}\u00b7Ex(D), deriving the enhancement of Ex(G) from the condition of Im{\u03c3(D)}\u2009<\u2009Im{\u03c3(G)}\u2009<\u20090. For the practical realization, we also note that H-GGP modes can be obtained at any finite temperatures when the conductivity of each domain satisfies the condition of Im{\u03c3(D)}\u2009<\u2009Im{\u03c3(G)}\u2009<\u20090\u2009<\u2009Im{\u03c3(M)}, as shown in f\u2009=\u2009100\u2009THz.w\u2009=\u20090) corresponds to the \u03c3(M)\u2009\u2212\u2009\u03c3(D) 1D-SPP system with the effective mode indexneff\u2009=\u2009Re{q}/k0 \u2248 2\u00b7(3/2)1/2\u00b7\u03b5\u03b50\u00b7c/(Im{\u03c3(M)}\u2009+\u2009Im{\u03c3(D)}), while the H-GGP system with infinite w is converged to the \u03c3(M)\u2009\u2212\u2009\u03c3(G) 1D-SPP system. The effective mode index (de index and the de index of the Hw case is converged to that of the \u03c3(M)\u2009\u2212\u2009\u03c3(G) 1D-SPP system, smaller w cases exhibit the intensity profiles focused of the graphene gap. Such a distinct in-plane intensity distribution imposes the unique property on out-of-plane confinement, in terms of the light-graphene overlap factor \u03c1\u2009=\u2009\u222b\u222bgraphene|E|2\u00b7dS/\u222b\u222b|E|2\u00b7dS: the concentration of electromagnetic fields on graphene. \u03c1 as a function of the gap width w, which demonstrates the improved light-graphene overlap for the structures with apparent field concentration on the gap (0\u2009<\u2009w\u2009<\u200940\u2009nm). We note that the H-GGP mode acquires much higher field concentration on the graphene layer (\u03c1\u2009=\u20092.07\u2009\u00d7\u200910\u22123 at w\u2009=\u20095\u2009nm), when compared to those of 1D-SPP modes (\u03c3(M)\u2009\u2212\u2009\u03c3(D) system of \u03c1\u2009=\u20091.81\u2009\u00d7\u200910\u22123 and \u03c3(M)\u2009\u2212\u2009\u03c3(G) system of \u03c1\u2009=\u20091.70\u2009\u00d7\u200910\u22123) and the GEP mode (\u03c1\u2009=\u20090.728\u2009\u00d7\u200910\u22123).Most importantly, there exist differentiated features of the H-GGP mode when compared to 1D-SPP modes, as shown in \u03a9(M))\u22121\u2009=\u20094\u2009+\u2009\u0394\u03a9\u22121, (\u03a9(G))\u22121\u2009=\u20090.54\u2009+\u2009\u0394\u03a9\u22121, and (\u03a9(D))\u22121\u2009=\u20090.5002\u2009+\u2009\u0394\u03a9\u22121, with the spatially global (wmax modulation width) modulation range. As seen, the effective mode index of the H-GGP mode can be controlled with an order of smaller modulation of \u0394\u03a9\u22121 when compared to the GEP mode. The H-GGP mode also provides more efficient regime of \u0394\u03a9\u22121 for controlling effective index compared to 1D-SPP modes (\u0394\u03a9\u22121\u2009\u2264\u20090.015). Such improved efficiency is more apparent for the case of the finite modulation region for \u0394\u03a9\u22121 (dotted lines in wmax modulation width around the graphene gap), due to the improved transverse localization of the H-GGP mode will be required for increasing excitation efficiency, for example, by applying adiabatic procedure or matched layers.We demonstrated the existence of low-dimensional gap plasmon modes on graphene, which supports large light-graphene overlap factor. The system with spatially-varying chemical potential (or doping level) for H-GGP modes can be realized by several existing schemes such as electric field bias332137How to cite this article: Kim, Y. et al. Low-dimensional gap plasmons for enhanced light-graphene interactions. Sci. Rep.7, 43333; doi: 10.1038/srep43333 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations."} +{"text": "In the crystal, supra\u00admolecular layers sustained by C\u2014H\u22efO, \u03c0\u2013\u03c0, C\u2014H\u22ef\u03c0(arene) and C\u2014H\u22ef\u03c0(chelate ring) inter\u00adactions are formed.A highly distorted penta\u00adcoordinated C 3)2(C28H32N2O4)], features a distorted SnC2NO2 coordination geometry almost inter\u00admediate between ideal trigonal\u2013bipyramidal and square-pyramidal. The dianionic Schiff base ligand coordinates in a tridentate fashion via two alkoxide O and hydrazinyl N atoms; an intra\u00admolecular hy\u00addroxy-O\u2014H\u22efN(hydrazin\u00adyl) hydrogen bond is noted. The alk\u00adoxy chain has an all-trans conformation, and to the first approximation, the mol\u00adecule has local mirror symmetry relating the two Sn-bound methyl groups. Supra\u00admolecular layers sustained by imine-C\u2014H\u22efO(hy\u00addroxy), \u03c0\u2013\u03c0 [between dec\u00adyloxy-substituted benzene rings with an inter-centroid separation of 3.7724\u2005(13)\u2005\u00c5], C\u2014H\u22ef\u03c0(arene) and C\u2014H\u22ef\u03c0(chelate ring) inter\u00adactions are formed in the crystal; layers stack along the c axis with no directional inter\u00adactions between them. The presence of C\u2014H\u22ef\u03c0(chelate ring) inter\u00adactions in the crystal is clearly evident from an analysis of the calculated Hirshfeld surface.The title diorganotin compound, [Sn(CH The bond lengths involving the nitro\u00adgen atoms comprising the backbone of the chelate rings suggest some conjugation, i.e. N1\u2014C1, N1\u2014N2 and N2\u2014C12 are 1.317\u2005(3), 1.397\u2005(2) and 1.303\u2005(3)\u2005\u00c5, respectively. The 10 atoms of the fused-ring system appended to the five-membered chelate ring make a dihedral angle of 2.01\u2005(3)\u00b0 with the chelate ring, a conformation allowing the formation of an intra\u00admolecular hy\u00addroxy-O\u2014H\u22efN(hydrazin\u00adyl) hydrogen bond to close an S(6) loop, Table\u00a02trans conformation with the range of torsion angles being \u2212174.96\u2005(18)\u00b0, for C21\u2014C22\u2014C23\u2014C24, to 179.79\u2005(19)\u00b0, for C25\u2014C26\u2014C27\u2014C28. Indeed, the r.m.s. deviation for the least-squares plane through all non-hydrogen atoms except the Sn-bound methyl groups is relatively small at 0.1179\u2005\u00c5, with maximum deviations being for the terminal methyl group of the alk\u00adoxy chain, i.e. 0.296\u2005(2)\u2005\u00c5, and a central methyl\u00adene-C22 atom, i.e. 0.194\u2005(2)\u2005\u00c5. Hence, to a first approximation, the mol\u00adecule has mirror symmetry, relating the two Sn-bound methyl groups.The five-membered, SnON3N2CH}2 synthon is formed, which encapsulates two six-membered {\u22efHOC3N} synthons formed by the intra\u00admolecular hy\u00addroxy-O\u2014H\u22efN(hydrazin\u00adyl) hydrogen bonding mentioned above, Fig.\u00a02a. Centrosymmetrically related dimeric aggregates are linked via \u03c0\u2013\u03c0 inter\u00adactions between dec\u00adyloxy-substituted benzene rings . The remaining inter\u00adactions are of the type C\u2014H\u22ef\u03c0 and involve methyl\u00adene-C\u2014H exclusively. While two of the inter\u00adactions have benzene rings as acceptors, the other two have chelate rings as acceptors, i.e. are of the type C\u2014H\u22ef\u03c0(chelate), a phenomenon gaining increasing attention hydrogen bond, the hy\u00addroxy-O atom accepts an inter\u00adaction from a centrosymmetrically-related imine-H atom, Table\u00a02et al., 2017dnorm, in the range \u22120.053 to + 1.621 au, Fig.\u00a033N2CH}2 synthon as discussed in the previous section; these are also viewed as blue and red regions near the H and O atoms on the Hirshfeld surface mapped over electrostatic potential (over the range \u00b1 0.075 au), Fig.\u00a04de mapped Hirshfeld surface, Fig.\u00a05A\u22ef\u03c0(C13\u2013C18) contacts at 1\u00a0\u2212\u00a0x, \u2212y, 1\u00a0\u2212\u00a0z and their reciprocal contacts, i.e. \u03c0\u22efH\u2014C, are represented with blue and white dotted lines, respectively, in Fig.\u00a06a. The other C\u2014H\u22ef\u03c0 contacts involving benzene rings and \u03c0\u2013\u03c0 stacking inter\u00adactions at 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z are illustrated in Fig.\u00a06b.The Hirshfeld surface analysis for (I)et al., 2007a\u2013f; their relative contributions are summarized qu\u00adanti\u00adtatively in Table\u00a03i.e. 97.0%.The overall two-dimensional fingerprint plot and those delineated into H\u22efH, C\u22efH/H\u22efC, O\u22efH/H\u22efO, N\u22efH/H\u22efN and C\u22efC contacts inter\u00adactions, Table\u00a02c and e, as their ring centroids benzene rings, and appears as an arrow-like distribution of points around de\u00a0=\u00a0di\u00a0\u223c\u00a01.9\u2005\u00c5 in Fig.\u00a07f. The other contacts, having low percentage contribution to the surface, are likely to have a negligible effect on the mol\u00adecular packing.A pair of very short peaks at et al., 2016Chemical context). Two di\u00admethyl\u00adtin structures are available with identical ligands apart from having a substituent in the 5-position, i.e. chloride According to a search of the crystallographic literature -3-hy\u00addroxy-2-napthohydrazide and tri\u00adethyl\u00adamine in ethyl acetate (25\u2005ml) were added to di\u00admethyl\u00adtin dichloride in ethyl acetate (10\u2005ml). The resulting mixture was stirred and refluxed for 3\u2005h. The filtrate was evaporated until a precipitate was obtained. The precipitate was recrystallized from di\u00adchloro\u00admethane:di\u00admethyl\u00adformamide (1:1), and yellow prismatic crystals suitable for X-ray crystallographic studies were obtained from the slow evaporation of the filtrate. Yield: 0.366\u2005g, 60%; M.p.: 507\u2013508\u2005K. IR (cm\u22121): 3162(br), 1633(s), 1597(s), 1169(s) cm\u22121. 1H NMR (in DMSO-d6): \u03b4 11.34 , 8.57 , 6.25\u20136.40, 7.07\u20137.20 , 8.47 , 3.96 , 1.28-1.82 , 0.91, , 0.89 .Uiso(H) set to 1.2\u20131.5Ueq(C). The oxygen-bound H atom was located in a difference Fourier map but was refined with a distance restraint of O\u2014H = 0.84\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.5Ueq(O). The maximum and minimum residual electron density peaks of 0.80 and 1.32\u2005e\u2005\u00c5\u22123 were located 0.42 and 0.83\u2005\u00c5, respectively, from the H23B and Sn atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989017002365/hb7655sup1.cifCrystal structure: contains datablock(s) . DOI: 10.1107/S2056989017002365/hb7655Isup2.hklStructure factors: contains datablock(s) I. DOI: 1532445CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The reaction of manganese dichloride and iron dichloride with the ligand 5,6-bis\u00ad(pyridin-2-yl)pyrazine-2,3-di\u00adcarb\u00adoxy\u00adlic acid leads to the formation of isotypic binuclear complexes which have a cage-like structure. 4N1,O2,N6:O3;\u03ba4O3:N1,O2,N6-bis\u00ad[di\u00adaqua\u00admanganese(II)] tetra\u00adhydrate, [Mn2(C16H8N4O4)2(H2O)4]\u00b74H2O, (I), and bis\u00ad-\u03ba4N1,O2,N6:O3;\u03ba4O3:N1,O2,N6-bis\u00ad[di\u00adaqua\u00adiron(II)] tetrahydrate, [Fe2(C16H8N4O4)2(H2O)4]\u00b74H2O, (II), are, respectively, the mangan\u00adese(II) and iron(II) complexes of the ligand 5,6-bis\u00ad(pyridin-2-yl)-pyrazine-2,3-di\u00adcarb\u00adoxy\u00adlic acid. The complete mol\u00adecule of each complex is generated by inversion symmetry. Each metal ion is coordinated by a pyrazine N atom, a pyridine N atom, two carboxyl\u00adate O atoms, one of which is bridging, and two water O atoms. The metal atoms have MN2O4 coordination geometries and the complexes have a cage-like structure. In the crystals of both compounds, the complexes are linked by O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds involving the coordinating water mol\u00adecules, forming chains along [100]. These chains are linked by O\u2014H\u22efO hydrogen bonds involving the non-coordinating water mol\u00adecules, forming layers parallel to (011). The layers are linked by pairs of C\u2014H\u22efO hydrogen bonds and offset \u03c0\u2013\u03c0 inter\u00adactions, so forming a hydrogen-bonded three-dimensional framework.The title isotypic complexes, bis\u00ad-\u03ba On exposure to air, this compound loses the solvent of crystallization and four water mol\u00adecules, transforming into a polymeric two-dimensional network structure pyrazine-2,3-di\u00adcarb\u00adoxy\u00adlic acid , and by two carboxyl\u00adate O atoms, O1 and O3i . Hence, the ligand coordinates to the metal atoms in a tridentate and a monodentate (O) manner. Atom O3 is bridg\u00ading, so leading to the formation of a cage-like complex situated about a centre of inversion; illustrated in Fig.\u00a03II complex, (II)ica 6.58\u2005\u00c5, while the Fe1\u22efFe1i distance is ca 6.50\u2005\u00c5. Selected bond lengths and angles for compounds (I)The complete mol\u00adecules of complexes (I)pyrazine, Mn1\u2014N1, bond length is 2.242\u2005(3)\u2005\u00c5, which is shorter than the Mn\u2014Npyridine, Mn1\u2014N3, bond length of 2.311\u2005(3)\u2005\u00c5. The Mn1\u2014Owater bond lengths [2.141\u2005(3) and 2.148\u2005(3)\u2005\u00c5] are similar to the Mn\u2014Ocarboxyl\u00adate, Mn1\u2014O3i, bond length of 2.139\u2005(2)\u2005\u00c5, while distance Mn1\u2014O1 is longer at 2.228\u2005(2)\u2005\u00c5.In complex (I)II two-dimensional coordination polymer involving ligand 2LH, mentioned above. The Fe\u2014Npyrazine bond length, Fe1\u2014N1, is 2.126\u2005(2)\u2005\u00c5, which is slightly shorter than the Fe\u2014Npyridine, Fe1\u2014N3, bond length of 2.205\u2005(3)\u2005\u00c5. The Fe1\u2014Owater bond lengths [2.115\u2005(2) and 2.066\u2005(2)\u2005\u00c5] are similar to the Fe1\u2014Ocarboxyl\u00adate bond lengths [2.131\u2005(2) and 2.139\u2005(2)\u2005\u00c5].In complex (II)W,O3i in the equatorial plane and atoms O2W and N1 in the apical positions with an O2W\u2014Mn1\u2014N1 bond angle of 163.62\u2005(11) \u00b0 (Table\u00a01W\u2014Fe1\u2014N1 bond angle of 165.15\u2005(10)\u00b0 (Table\u00a02viz. 57.28\u2005(17)\u00b0.The geometry of the sixfold coordinated metal atoms can best be described as a distorted octa\u00adhedron, with atoms O1,N3,O1\u00b0 Table\u00a01, and an \u00b0 Table\u00a02. The cooW and O2W), forming chains along [100]; illustrated in Fig.\u00a04W and O4W), forming layers parallel to the bc plane, as illustrated in Fig.\u00a05Cg\u22efCgii = 3.671\u2005(4)\u2005\u00c5 in (I)Cg is the centroid of the ring N3/C5\u2013C9; symmetry code: (ii) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01], link the layers, forming a three-dimensional framework; illus\u00adtrated in Fig.\u00a06Details of the hydrogen-bonding inter\u00adactions in the crystals of both compounds, are given in Table\u00a03fs Fig.\u00a05. Pairs oet al., 20162LH, and its dimethyl ester, gave eight hits. Some of these structures have been mentioned in the Chemical context above. In the case of (I)N,N,O) and monodentate (O) manner. This coordination mode of 2LH is the same as that observed in the CdII two-dimensional coordination polymer has been reported previously pyrazine-2,3-di\u00adcarb\u00adoxy\u00adlic acid (Synthesis of compound (I): 2LH was added in solid form to an aqueous solution (15\u2005ml) of MnCl2\u00b74H2O . The yellow solution immediately obtained was stirred for 10\u2005min at room temperature, filtered and the filtrate allowed to slowly evaporate. After two weeks orange\u2013yellow rod-like crystals were obtained. They were separated by filtration and dried in air . Selected IR bands : \u03bd 3226, 3080(w), 1636(s), 1598(vs), 1545(w), 1475(m), 1440(m), 1410(w), 1366(s), 1348(s), 1301(w), 1275(w), 1170(m), 1126(m), 1007(w), 954(w), 850(w), 790(m), 562(m).Synthesis of compound (II): A degassed aqueous solution (20\u2005ml) of 2LH was treated with FeCl2\u00b74H2O . The violet solution immediately obtained was stirred under N2 at 343\u2005K for 1\u2005h, filtered and the filtrate allowed to slowly evaporate. After two months deep-violet block-like crystals were obtained. They were separated by filtration and air dried . Precipitation of small amounts of iron(III) hydroxide accompanied the formation of the crystals. Selected IR bands : \u03bd 3477, 3291, 3078(w), 1640(s), 1593(vs), 1545(w), 1475(m), 1440(m), 1405(w), 1359(m), 1300(w), 1286(w), 1269(w), 1172(m), 1124(m), 1008(w), 954(w), 847(w), 789(m), 772(w), 677(w), 565(m), 549(w), 494(m) %.Uiso(H) = 1.2Ueq(C). Intensity data for (I)Rint = 0, and as no suitable \u03c8-scans could be measured no absorption correction was applied. For compound (II)Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989016014055/pj2034sup1.cifCrystal structure: contains datablock(s) I, II, Global. DOI: 10.1107/S2056989016014055/pj2034Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016014055/pj2034IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1502352, 1502351CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Single-molecule magnet (SMM) properties of these four complexes were observed\u00a0to be dependent on the ROH lattice solvent molecule. There is an interesting magneto-structural correlation: the larger the R group, the higher the energy barrier. For the first time, the\u00a0solvatomagnetic effect is\u00a0used for the\u00a0continuous fine adjustment\u00a0of the energy barrier of 0D molecular nanomagnets. Additionally, [Cu3Dy2(H3L)2(OAc)2(hfac)4]\u22192MeOH (5), an\u00a0analogue of [Cu3Tb2(H3L)2(OAc)2(hfac)4]\u22192MeOH (1), is also reported for comparison.Solvents play important roles in our lives, they are also of interest in molecular materials, especially for molecular magnets. The solvatomagnetic effect is generally used for trigger and/or regulation of magnetic properties in molecule-based\u00a0systems, however, molecular nanomagnets showing solvatomagnetic effects are very difficult to obtain. Here we report four 3d-4f heterometallic cluster complexes containing ROH lattice solvent molecules, [Cu The solvatomagnetic effect is very interesting because solvent molecules can be used for trigger and/or regulation of magnetic properties while the molecular magnetic structure is always maintained. Therefore, molecular magnets showing solvatomagnetic effects can be used as molecule devices, molecular switches and/or molecular sensors. Naturally, solvatomagnetic effects are\u00a0often found\u00a0in porous metal-organic frameworks (MOFs) in which solvent molecules are guest molecules9, while low-dimensional systems with solvatomagnetic effects are more difficult to obtain due to the\u00a0lack of pores. Recently, we found a chain-like azido-bridged manganese(III) coordination polymer showing both solvatomagnetic effect and spin-glass behaviour10. In studies of single molecule magnets (SMMs)11, we also hope to explore SMM systems with solvatomagnetic effects. However, it is a great challenging task because most SMMs\u00a0reported are concentrated on zero-dimensional (0 D) cluster or mononuclear systems.Solvents, especially water, are critical to the origins of life, and they have penetrated into all aspects of human life. Besides as reaction mediums and extracting agents, chemical solvents are also of interest in molecular materials. For example, in the field of molecular magnets they can\u00a0be utilized as the\u00a0terminal ligand to complete the coordination configuration17; and the energy barrier leading to magnetic bistability and slow magnetic relaxation is a pivotal parameter. Therefore, except enhancing the relaxation energy barrier and increasing the blocking temperature21, tuning the relaxation energy barrier is another important target in the molecular nanomagnet field26. Surprisingly, systematic studies\u00a0of SMMs with the same magnetic structure are\u00a0still rare, however, some factors such as the\u00a0electron-withdrawing effect27, the\u00a0electrostatic potential of the key coordination atom28 have been observed to be able to modulate SMMs\u2019 energy barriers recently. Regarding structures and magnetic properties may be affected by a small change of circumstance, solvent molecules may also be used to adjust SMMs\u2019 properties. To the best of our knowledge, a direct correlation between energy barriers and different lattice solvent molecules of 0D molecular nanomagnets has never been documented, though a 3D Dy(III) MOF-type SMM was found to show an obvious solvatomagnetic effect in 201529, and guest-dependent single-ion magnet behaviours were observed in a 2D cobalt(II) coordination polymer\u00a0in 201630. Herein we describe the lattice-solvent effect of ROH molecules on the energy barrier of 0D SMMs with the same magnetic structure [Cu3Tb2(H3L)2(OAc)2(hfac)4] {H6L\u2009=\u20091,3-Bis[tris(hydroxymethyl)methylamino]propane, Fig.\u00a0\u2212\u2009=\u2009hexafluoroacetylacetonate}. Fine adjustment of the energy barrier in this [Cu3Tb2(H3L)2(OAc)2(hfac)4] SMM system was achieved by changing the\u00a0ROH lattice solvent molecule. A similar [Cu3Dy2(H3L)2(OAc)2(hfac)4] SMM system was also studied, but only the crystal structure of [Cu3Dy2(H3L)2(OAc)2(hfac)4]\u22192MeOH (5) was successfully\u00a0solved; complex 5 also exhibits slow magnetic relaxation under zero dc field, with the energy barrier of 30.0\u2009K, a relatively high value for reported relaxation barriers of the Cu-Dy heterometallic SMMs.It is well known that SMMs are of great potential for technological applications in high-density information storage, quantum computing and spintronics6L), an universal ligand due to flexible polydentate coordination sites, has been used to bind\u00a0not only 3d transition metal ions32 but also 4\u2009f lanthanide metal ions33. Furthermore, it can also be utilized to construct 3d-4f heterometallic complexes34. Recently, Murrie et al. reported a series of 3d-4f complexes formulated as {Ln2Cu3(H3L)2Xn} 35; they found that changing\u00a0the auxiliary ligand OAc\u2212 through\u00a0NO3\u2212 may lead to a remarkable improvement of the energy barrier of {Tb2Cu3(H3L)2Xn} (X\u2009=\u2009OAc\u2212 and NO3\u2212) complexes, which suggests that the anion co-ligand has\u00a0a great impact on the energy barrier of {Tb2Cu3(H3L)2Xn} SMMs. In the recent process of pursuing new SMMs, we observed that using Ln(OAc)(hfac)2(H2O)2 as the lanthanide (III) salt source may lead mixed co-ligands OAc\u2212 and hfac\u2212 into 3d-4f heterometallic clusters effectively36. Therefore, we adopted this synthesis strategy to obtain the [Cu3Tb2(H3L)2(OAc)2(hfac)4] SMM with different ROH lattice solvent molecules , in which not only the OAc\u2212 anion but also the hfac\u2212 anion are co-ligands. Notably, our synthetic procedures were\u00a0completed at room temperature rather than at 60\u2009\u00b0C used by Murrie group35. Products\u00a0using methanol, ethanol and isopropyl alcohol as reaction solvents were [Cu3Tb2(H3L)2(OAc)2(hfac)4]\u22192MeOH (1), [Cu3Tb2(H3L)2(OAc)2(hfac)4]\u22192EtOH (2) and [Cu3Tb2(H3L)2(OAc)2(hfac)4]\u22192iso-C3H7OH (3), respectively; while [Cu3Tb2(H3L)2(OAc)2(hfac)4]\u22192H2O (4) was quantitatively transformed from complex 1 by taking place of methanol molecules with water molecules. In order to yield [Cu3Tb2(H3L)2(OAc)2(hfac)4] SMMs with larger ROH lattice solvent molecules, other ROH solvents such as isobutyl alcohol, n-butyl alcohol and isoamyl alcohol were also used instead of methanol for 1, but no any crystalline products could be obtained. Furthermore, the [Cu3Dy2(H3L)2(OAc)2(hfac)4] SMM system was also explored, but only the crystal structure of [Cu3Dy2(H3L)2(OAc)2(hfac)4]\u22192MeOH (5) was successfully\u00a0solved, the crystal structure of [Cu3Dy2(H3L)2(OAc)2(hfac)4] SMMs with other lattice solvent molecules could not be obtained due to the severe\u00a0twinning phenomenon.Bis-tris propane (H3Tb2(H3L)2(OAc)2(hfac)4]\u22192ROH SMMs have the main structure [Cu3Tb2(H3L)2(OAc)2(hfac)4] 2(OAc)2(hfac)4]\u22192MeOH (1) is chose to be described in detail. In the main structure [Cu3Tb2(H3L)2(OAc)2(hfac)4], a {Cu3(H3L)2} linear unit is formed through bridging two terminal {Cu(H3L)}\u2212 fragments using\u00a0a central Cu2+ ion, then two Tb3+ ions link to this {Cu3(H3L)2} linear unit in the opposite direction, in which each Tb3+ ion connects with the central Cu2+ ion and one external Cu2+ ion through sharing one \u03bc3-O atom\u00a0and one \u03bc-O atom from one H3L3\u2212 ligand, and one \u03bc3-O atom from the other H3L3\u2212 ligand 2Xn}35. The eight-coordinate sphere of each Tb3+ ion is finally completed by two hfac\u2212 anions and one OAc\u2212 anion. Shape software37 was adopted to calculate the Tb(III) coordination polyhedron, giving a triangular dodecahedron as the most likely configuration for complex 1, and the deviation value from the ideal D2d symmetry is 1.015 coordination polyhedra of complexes 2\u20134 using Shape software37 are listed in Tables\u00a0All SMM system.The direct current (dc) variable-temperature magnetic susceptibility\u00a0of complexes eld Fig.\u00a0. The roo 24.91\u2009cm\u2009K mol\u22121,\u03c7\u2032, Fig.\u00a0\u03c7\u2032\u2032, Fig.\u00a01\u20134 are frequency-dependent in zero dc field, indicating slow magnetic relaxation typical for SMMs. Such thermally induced relaxation was fitted with the Arrhenius law, \u03c4\u2009=\u2009\u03c40exp(Ueff/kT), extracting Ueff/k values of 30.0(0.4) K for 1, 32.4(0.2) K for 2, 33.1(0.7) K for 3 and 25.7(0.2) K for 4 as well as \u03c40 values of 3.7(0.2)\u2009\u00d7\u200910\u22128 s for 1, 6.2(0.1)\u2009\u00d7\u200910\u22129 s for 2, 2.6(0.3)\u2009\u00d7\u200910\u22128 s for 3 and 2.3(0.1)\u2009\u00d7\u200910\u22128 s for 4 13. A comparison of the effective barrier value for complexes 1\u20134 with the R group of the ROH lattice solvent molecules reveals an important magneto-structural correlation for this [Cu3Tb2(H3L)2(OAc)2(hfac)4] SMM system: The larger the R group in ROH, the higher the energy barrier of the [Cu3Tb2(H3L)2(OAc)2(hfac)4]\u22192ROH SMM 2[Tb2Cu3(H3L)2(NO3)7(CH3OH)2](NO3) (36\u2009K)35; the Ueff/k value of 1 is also remarkable, which is comparable with that of [Cu22Tb2(N3)6]\u00b72CH3OH 38. In many cases44, a dc field is necessary for 3d-4f heterometallic complexes to display magnetic relaxation because of the\u00a0obvious quantum-tunnelling effects.The solvatomagnetic effect could also be detected by alternating current (ac) magnetic susceptibility investigations. Both the in-phase \u03c7\u2032, Fig.\u00a0, SI and r 4 Fig.\u00a0. All fouSMM Fig.\u00a0. It is n3Tb2(H3L)2Xn] system is not because of single-ion behaviours, and both the Cu\u00b7\u00b7\u00b7Cu and Cu\u00b7\u00b7\u00b7Tb ferromagnetic interactions maybe quench the tunnel splitting, which are similar to acting as an internal applied field, inducing to zero-field SMM behaviours35. Nevertheless, the difference of the Tb3+ coordination configurations has\u00a0influence on the SMM characteristics35. Owing to great difficulty for theoretical calculation and comparison of the Cu\u00b7\u00b7\u00b7Cu and Cu\u00b7\u00b7\u00b7Tb ferromagnetic couplings35, we tried to make a magneto-structural correlation for complexes 1\u20134 using the deviation value from the ideal D2d symmetry of the\u00a0biaugmented trigonal prism for the Tb3+ ion and the intermolecular distance as two main structural parameters. As shown in Table\u00a03+ ions is\u00a0closer to the biaugmented trigonal prism from 1 to 3, the corresponding energy barrier value becomes larger from 1 to 3, indicating the biaugmented trigonal prism configuration in the [Cu3Tb2(H3L)2(OAc)2(hfac)4] SMM system is the dominant configuration; but 4 is a bit unusual, its deviation value (1.735) is comparable with that of 1 (1.756), which suggests that\u00a0other structural factors such as intermolecular distances need to be considered; as shown in Table\u00a0central\u2026Cucentral separation), the higher the energy barrier; which is in line with the magneto-structural correlation using the R group itself, because larger ROH lattice solvent molecules may enhance intermolecular distances correspondingly.Simplified theoretical investigations by Murrie group suggested that the magnetic bistability in the (NO3)35, and the ferromagnetic coupling obviously exists between the Cu2+ ion and the Dy3+ ion as well as among the Cu2+ ions, similar to that observed in [Gd2Cu3(H3L)2(CH3COO)6]\u00b7THF\u00b73H2O by Murrie group35.The l\u22121 Fig.\u00a0, which i5 are similar to those of complexes 1\u20134. Under zero dc field, the appearance of frequency-dependent \u03c7\u2032 K and \u03c40\u2009=\u20099.7(0.1)\u2009\u00d7\u200910\u22129 s 2{Dy2Cu3(H3L)2(NO3)7(CH3OH)2](NO3) [23.9(0.1) K], whose \u03c7\u2033 signals even do not appear peaks in zero dc field35. Notably, this Ueff/k value is the third high value for the Cu-Dy heterometallic SMMs, after 47\u2009K of [{Dy(hfac)3}2{Cu(dpk)2}] (dpk\u2212\u2009=\u2009di-2-pyridyl ketoximate)48 and 41.6\u2009K of [Cu4Dy4(vanox)6(Hvanox)2(NO3)4(\u03bc-HOMe)2]\u00b76MeOH 49. Furthermore, this Ueff/k value is remarkable larger than those of the Cu-Dy heterometallic SMMs with higher nucleus (<20\u2009K)51. Additionally, the parameter \u03a6 value of 0.16 for 5 supports the SMM nature too.The magnetization dynamics of compound \u03c7\u2032 Fig.\u00a0, SI and als Fig.\u00a0 indicate9 s Fig.\u00a0. The ene5 revealed that the \u03c7\u2033 signals of 5 are temperature-dependent methylamino]propane ligand (H6L). The ROH lattice solvent molecules in the [Cu3Tb2(H3L)2(OAc)2(hfac)4] SMM system have great influences on the energy barrier; the larger the R group, the higher the energy barrier. We predict that the larger ROH molecule may enlarge the intermolecular distance and can help to change the coordination configuration of the Ln(III) ions through the hydrogen bonding interaction between the ROH lattice solvent molecule and the [Cu3Tb2(H3L)2(OAc)2(hfac)4] main-structural molecule. Our work demonstrates that solvatomagnetic effects can be used to\u00a0continuously fine-tune energy barriers in\u00a0SMMs. The discovery is bound to have significances in enhancing and turning energy barriers of molecular nanomagnets via chemical methods such as using lattice-solvent effects.In summary, a mixed OAcThe elemental analyses were measured on a Vario ELIII elemental analyser. The magnetic susceptibility measurements were carried out on a Quantum Design MPMS-XL5 SQUID magnetometer, and diamagnetic corrections were calculated from Pascal\u2019s constants of all components.6L (0.25\u2009mmol) and Cu(ClO4)2\u00b76H2O (0.375\u2009mmol) in 20\u2009mL of MeOH, was added Tb(OAc)(hfac)2(H2O)2 (0.15\u2009mmol), a blue solution was formed after being stirred for 10\u2009min; Et3N (0.75\u2009mmol) was then added dropwise, the resultant solution was stirred for 3\u2009h at room temperature and turned violet. Violet plate-like X-ray quality crystals were obtained through slow evaporation of the filtrate at room temperature over 1 week. Yield (25%). Anal. Calcd (%) for C48H64Cu3F24N4O26Tb2 (1) C 27.75; H 3.11; N 2.70. Found: C 27.80; H 3.14; N 2.67.To a mixture of H1 was followed, but using ethanol instead of methanol. Violet plate-like X-ray quality crystals were obtained through slow evaporation of the filtrate at room temperature over 10 days. Yield (27%). Anal. Calcd (%) for C50H68Cu3F24N4O26Tb2 (2): C 28.52; H 3.26; N 2.66. Found: C 28.55; H 3.29; N 2.63.The same synthetic procedure for complex 1 was followed, but using isopropyl alcohol instead of methanol. Violet plate-like X-ray quality crystals were obtained through slow evaporation of the filtrate at room temperature over 15 days. Yield (22%). Anal. Calcd (%) for C52H72Cu3F24N4O26Tb2 (3): C 29.27; H 3.40; N 2.63. Found: C 29.23; H 3.43; N 2.60.The same synthetic procedure for complex 1 was kept at 60\u2009\u00b0C for 6\u2009h, and then exposed on air for 24\u2009h. Violet plate-like X-ray quality crystals of 4 were obtained quantitatively. Anal. Calcd (%) for C46H60Cu3F24N4O26Tb2 (4): C 26.96; H 2.95; N 2.73. Found: C 27.02; H 2.99; N 2.69.Complex 1 was followed, but using Dy(OAc)(hfac)2(H2O)2 instead of Tb(OAc)(hfac)2(H2O)2. Violet plate-like X-ray quality crystals were obtained through slow evaporation of the filtrate at room temperature over 1 week. Yield (28%). Anal. Calcd (%) for C48H64Cu3Dy2F24N4O26 (5): C 27.66; H 3.09; N 2.69. Found: C 27.69; H 3.11; N 2.67.The same synthetic procedure for complex 3 of 1, 0.178\u2009\u00d7\u20090.063\u2009\u00d7\u20090.024\u2009mm3 of 2, 0.183\u2009\u00d7\u20090.125\u2009\u00d7\u20090.031\u2009mm3 of 3, 0.108\u2009\u00d7\u20090.067\u2009\u00d7\u20090.025\u2009mm3 of 4, and 0.134\u2009\u00d7\u20090.125\u2009\u00d7\u20090.027\u2009mm3 of 5 was picked out to mount on a Bruker SMART APEX-CCD diffractometer with Mo-K\u03b1 radiation (\u03bb\u2009=\u20090.71073\u2009\u00c5) for data collection at 173(2) K. Empirical absorption corrections from \u03c6 and \u03c9 scan were applied. Cell parameters were calculated by the global refinement of the positions of all collected reflections for five complexes. The structures were solved by direct methods and refined by a full matrix least-squares technique based on F2 using with the SHELX-2014 program package. All hydrogen atoms were set in calculated positions and refined as riding atoms, and all non-hydrogen atoms were refined anisotropically. CCDC 1574978\u20131574982 contain the supplementary crystallographic data, which can be obtained free of charge from the Cambridge Crystallographic Data Centre via www.ccdc.cam.ac.uk/data_request/cif.A single crystal with dimensions 0.261\u2009\u00d7\u20090.093\u2009\u00d7\u20090.025\u2009mm1: P\u22121, a\u2009=\u200910.086(2)\u2009\u00c5, b\u2009=\u200912.463(3)\u2009\u00c5, c\u2009=\u200915.594(3)\u2009\u00c5, \u03b1\u2009=\u2009104.67(3)\u00b0, \u03b2\u2009=\u200994.07(3)\u00b0, \u03b3\u2009=\u2009108.97(3)\u00b0, V\u2009=\u20091767.9(7)\u2009\u00c53, Mr\u2009=\u20092077.49, Dc\u2009=\u20091.951\u2009g\u2009cm\u22123, Z\u2009=\u20091, R1\u2009=\u20090.0366 (I\u2009>\u20092\u03c3(I)), wR2\u2009=\u20090.0878 (I\u2009>\u20092\u03c3(I)), S\u2009=\u20091.080.Crystal data for 2: P\u22121, a\u2009=\u200910.243(2)\u2009\u00c5, b\u2009=\u200912.469(3)\u2009\u00c5, c\u2009=\u200915.602(3)\u2009\u00c5, \u03b1\u2009=\u2009101.71(3)\u00b0, \u03b2\u2009=\u200996.50(3)\u00b0, \u03b3\u2009=\u2009110.00(3)\u00b0, V\u2009=\u20091797.2(6)\u2009\u00c53, Mr\u2009=\u20092105.54, Dc\u2009=\u20091.945\u2009g\u2009cm\u22123, Z\u2009=\u20091, R1\u2009=\u20090.0498 (I\u2009>\u20092\u03c3(I)), wR2\u2009=\u20090.1085 (I\u2009>\u20092\u03c3(I)), S\u2009=\u20091.122.Crystal data for 3: P\u22121, a\u2009=\u200910.309(2)\u2009\u00c5, b\u2009=\u200912.473(3)\u2009\u00c5, c\u2009=\u200915.677(3)\u2009\u00c5, \u03b1\u2009=\u2009101.80(3)\u00b0, \u03b2\u2009=\u200996.96(3)\u00b0, \u03b3\u2009=\u2009110.19(3)\u00b0, V\u2009=\u20091811.4(7)\u2009\u00c53, Mr\u2009=\u20092133.58, Dc\u2009=\u20091.956\u2009g\u2009cm\u22123, Z\u2009=\u20091, R1\u2009=\u20090.0345 (I\u2009>\u20092\u03c3(I)), wR2\u2009=\u20090.0797 (I\u2009>\u20092\u03c3(I)), S\u2009=\u20091.084.Crystal data for 4: P\u22121, a\u2009=\u200910.042(2)\u2009\u00c5, b\u2009=\u200912.480(3)\u2009\u00c5, c\u2009=\u200915.819(3)\u2009\u00c5, \u03b1\u2009=\u2009107.08(3)\u00b0, \u03b2\u2009=\u200999.23(3)\u00b0, \u03b3\u2009=\u2009109.83(3)\u00b0, V\u2009=\u20091706.2(7)\u2009\u00c53, Mr\u2009=\u20092049.44, Dc\u2009=\u20091.995\u2009g\u2009cm\u22123, Z\u2009=\u20091, R1\u2009=\u20090.0488 (I\u2009>\u20092\u03c3(I)), wR2\u2009=\u20090.0945 (I\u2009>\u20092\u03c3(I)), S\u2009=\u20091.153.Crystal data for 5: P\u22121, a\u2009=\u200910.085(2)\u2009\u00c5, b\u2009=\u200912.427(3)\u2009\u00c5, c\u2009=\u200915.581(3)\u2009\u00c5, \u03b1\u2009=\u2009104.59(3)\u00b0, \u03b2\u2009=\u200994.21(3)\u00b0, \u03b3\u2009=\u2009108.86(3)\u00b0, V\u2009=\u20091762.5(7)\u2009\u00c53, Mr\u2009=\u20092084.65, Dc\u2009=\u20091.964\u2009g\u2009cm\u22123, Z\u2009=\u20091, R1\u2009=\u20090.0321 (I\u2009>\u20092\u03c3(I)), wR2\u2009=\u20090.0762 (I\u2009>\u20092\u03c3(I)), S\u2009=\u20091.074.Crystal data for Supplementary Information"} +{"text": "Only a weak directional inter\u00adaction of the C\u2014H\u22efO type combines mol\u00adecules in infinite chains running along the 23H26O5 or CH2=C(CH3)\u2014C(O)O\u2014C6H4\u2014O(O)C\u2014C6H4\u2014OC6H13, has been determined. The mol\u00adecule is non-planar and the dihedral angle between the phenyl rings is 50.72\u2005(4)\u00b0. The crystal packing differs from those typical for mesogenic compounds. Only a weak directional inter\u00adaction of the C\u2014H\u22efO type combines mol\u00adecules in endless chains running along the a axis.The structure of the title compound, C The structural studies of these compounds are of great inter\u00adest as these investigations make it possible to clarify the structure of the mesophase and propose a mechanism of phase transitions in a crystal-mesophase-isotropic system.Phenyl\u00adbenzoates bearing a rather long aliphatic substituent at the benzene ring are potentially mesogenic compounds. On melting, these compounds often form smectic or nematic phases. Cases where these compounds exhibit a monotropic mesomorphism, IIICr 367.7\u2005K Iso 350.6\u2005K IICr 349.9\u2005K ICr.In this work we performed an X-ray structural determination and DSC study of the title compound. According to DSC the compound is non\u2013mesomorphic, exhibiting three solid-state modifications: viz. benzene rings C8\u2013C13 (plane I) and C2\u2013C7 (plane II), ester groups C2/C1/O1/O2 (plane III) and O4/O5/C20/C21 (plane IV) and the hex\u00adyloxy group O3/C14\u2013C19 (plane V). The dihedral angles between the planes I/II, II/III, II/V, I/III and I/IV are 50.72\u2005(4), 4.84\u2005(5), 7.05\u2005(3), 52.82\u2005(4) and 55.50\u2005(5)\u00b0, respectively. According to the CSD Groom et al., 2016nHn+12 (n > 4), this substituent has an extended structure and its plane is nearly coplanar with the plane of the corresponding benzene ring.The unit cell contains one independent mol\u00adecule whose structure is shown in Fig.\u00a01et al., 2009et al., 2012et al., 2009n--n\u2032-tolyidines related mol\u00adecules may be considered to be weak hydrogen bonds I. DOI: 10.1107/S2056989017008568/rk2436Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017008568/rk2436Isup3.cmlSupporting information file. DOI: 1554948CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecule is V-shaped and possesses mirror symmetry; the mirror bis\u00adects the central benzene ring. There are two intra\u00admolecular O\u2014H\u22efN hydrogen bonds present forming S(6) ring motifs.The title Schiff base compound was synthesized 34H28N2O4, is generated by mirror symmetry, with the mirror bis\u00adecting the central benzene ring. It was synthesized via the condensation reaction of 1,2-di\u00adamine\u00adbenzene with 4-benz\u00adyloxy-2-hy\u00addroxy\u00adbenzaldehyde. The mol\u00adecule is V-shaped and there are two intra\u00admolecular O\u2014H\u22efN hydrogen bonds present forming S(6) ring motifs. The configuration about the C=N imine bonds is E. The central benzene ring makes dihedral angles of 41.9\u2005(2) and 43.6\u2005(2)\u00b0 with the phenol ring and the outer benz\u00adyloxy ring, respectively. The latter two rings are inclined to each other by 84.4\u2005(2)\u00b0. In the crystal, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming layers lying parallel to the ab plane. The Hirshfeld surface analysis and the two-dimensional fingerprint plots confirm the predominance of these inter\u00adactions in the crystal structure. The anti\u00adoxidant capacity of the compound was determined by the cupric reducing anti\u00adoxidant capacity (CUPRAC) process.The whole mol\u00adecule of the title Schiff base compound, C In view of this inter\u00adest we have synthesized the title compound, (I)1H NMR NMR spectrum reveals the presence of an imino group (N=CH) in the range \u03b4 = 8.5\u20138.7 p.p.m. The anti\u00adoxidant capacity of the compound was determined by the cupric reducing anti\u00adoxidant capacity (CUPRAC) process.Schiff base derivatives are a biologically versatile class of compounds possessing diverse activities, such as anti-oxidant (Haribabu viz. bonds C1\u2014C1i and C3\u2014C3i . In the mol\u00adecule there are two intra\u00admolecular O\u2014H\u22efN hydrogen bonds present (Table\u00a01S(6) ring motifs as shown in Fig.\u00a01E and the C4=N1 bond length is 1.278\u2005(6)\u2005\u00c5. The C3\u2014N1=C4 bond angles are less than 120\u00b0 [118.9\u2005(4)\u00b0], and the imine group has a C3\u2014N1\u2014C4\u2014C5 torsion angle of \u2212176.8\u2005(4)\u00b0. The mol\u00adecule is V-shaped and the two arms are non-planar; the central benzene ring forms dihedral angles of 41.9\u2005(2) and 43.6\u2005(2)\u00b0 with the phenol ring (C5-C10) and the outer benz\u00adyloxy ring (C12\u2013C17), respectively. The latter two rings are almost normal to each other, with a dihedral angle of 84.4\u2005(2)\u00b0.The mol\u00adecular structure of compound (I)t Table\u00a01, which fIn the crystal of (I)et al., 2007CrystalExplorer17 and H\u22efC/C\u22efH at 34.6% , followed by the H\u22efO/O\u22efH inter\u00adactions at 13.6% .The Hirshfeld surface analysis ]diphenol ethanol solvate (II) ]diphenol (III) ring motifs.A search of the Cambridge Structural Database the phenol rings are inclined to the central benzene ring by 53.9\u2005(3) and 4.0\u2005(2)\u00b0 and to each other by 49.9\u2005(2)\u00b0. In (III) the corresponding dihedral angles are 48.12\u2005(8), 21.44\u2005(8) and 47.70\u2005(8)\u00b0, while in (V) the corresponding dihedral angles are 58.29\u2005(12), 2.20\u2005(12) and 57.60\u2005(12)\u00b0. In compound (IV), that possesses twofold rotational symmetry with the twofold axis bis\u00adecting the central benzene ring, the phenol rings are inclined to the central benzene ring by 82.30\u2005(5)\u00b0 and to each other by 63.76\u2005(5)\u00b0. In the title compound, which possesses mirror symmetry, the corresponding dihedral angles are 41.9\u2005(2) and 68.9\u2005(2)\u00b0.o-phenyl\u00adenebis(nitrilo\u00admethyl\u00adidyne)]diphenolato)nickel(II) dihydrate methylyl\u00adidene]}di\u00adbenzene-1,3-diolato-\u03baO3)copper(II) methanol solvate II-Nc] (CUPRAC) method Nc2\u2013Cu+2 complex. Indeed, in the presence of an anti\u00adoxidant agent, the copper\u2013neocuproene complex is reduced and this reaction is qu\u00adanti\u00adfied spectrophotometrically at a wavelength of 450\u2005nm.The anti\u00adoxidant activity profile of the synthesized compound (I)0.50 = 15.03 \u00b1 1.50 for a 4\u2005mg dosage, compared to the results for buthylated toluene (BHT) [A0.50 = 8.97 \u00b1 3.94], used as a positive control (see Table\u00a02p < 0.05).According to the cupric ion reducing anti\u00adoxidant capacity assay, the title compound displayed activity with variable potency in all tested concentrations, because the percentage (%) inhibition in the CUPRAC assay is good [Ain vacuo. The residue was recrystallized from ethanol, yielding yellow block-like crystals of the title compound on slow evaporation of the solvent. The purity of the compound was characterized by its NMR spectrum . The azomethine proton appears in the 8.5\u20138.7 p.p.m. range, while the imine bond is characterized in the 13C RMN spectrum with the imine C and OH signals in the range 162.23\u2013163.34 p.p.m. 1H NMR: \u03b4 = 6.5\u20137.6 , \u03b4 = 13.7 , \u03b4 = 5.1 . 13C NMR: 70.15, 120.33, 127.30, 127.64, 128.26, 128.75, 142.32, 162.23, 163.33, 163.34.1,2-Di\u00adamine\u00adbenzene (0.027\u2005g) and 4-benz\u00adyloxy-2-hy\u00addroxy\u00adbenzaldehyde (0.1141\u2005g) in ethanol (15\u2005ml) were refluxed for 1\u2005h, then the solvent was evaporated Uiso(H) = 1.5Ueq(O). The C-bound H atoms were positioned geometrically (C\u2013H = 0.93\u20130.97\u2005\u00c5) and refined as riding with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018005832/su5438sup1.cifCrystal structure: contains datablock(s) Global, I. DOI: 10.1107/S2056989018005832/su5438Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018005832/su5438Isup3.cmlSupporting information file. DOI: 1837095CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular and crystal structure of the title Schiff base derivative is reported. The crystal packing depends on O\u2014H\u22efN hydrogen-bonds, augmented by C\u2014H\u22ef\u03c0 inter\u00adactions. 17H18N2O, the central carbon atom with the OH substituent and one of the (E)-benzyl\u00adidene\u00adamino substituents are disordered over two sets of sites with occupancies of 0.851\u2005(4) and 0.149\u2005(4). The relative positions of the two disorder components is equivalent to a rotation of approximately 60\u00b0 about the C\u2014N single bond. In the crystal, the mol\u00adecules are held together by O\u2014H\u22efN hydrogen bonds, forming simple C(5) chains along the b-axis direction. In addition, pairs of the chains are further aggregated by weak C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, C To gain more insight into the structural and spectroscopic properties of this potentially polydentate ligand, we report herein the mol\u00adecular structure of the title compound.During the last decades, inter\u00adest in Schiff bases and their complexes has been constant due to their extensive use for industrial purposes and also for their broad range of biological activities :0.149\u2005(4). The difference between the two conformers is reflected in the relative arrangement of the central spacer units. In the major disorder component, the torsion angle C3\u2014C4\u2014N2\u2014C5 is \u2212158.7\u2005(2)\u00b0 whereas the corresponding angle C3\u2032\u2014C4\u2032\u2014N2\u2032\u2014C5\u2032 in the minor component is \u221293.3\u2005(14)\u00b0. This translates to a rotation of approximately 60\u00b0 about the C4\u2014N2 bond. In the second, fully ordered, (E)-benzyl\u00adidene\u00adamino substituent, the equivalent torsion angles C1\u2014N1\u2014C2\u2014C3 and C1\u2014N1\u2014C2\u2014C3\u2032 are \u2212102.03\u2005(18)\u00b0 and \u221279.8\u2005(8)\u00b0, respectively.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01synclinal (-sc) alignment of the hydroxyl and imine nitro\u00adgen atoms around the N(imine)\u2014C\u2014C\u2014O(hydrox\u00adyl) bond indicate a slight loss of the sp2 character. The N1=C1 azomethine group is essentially co-planar with the attached benzene ring with an N1\u2014C1\u2014C11\u2014C12 torsion angle being 2.0\u2005(5)\u00b0. In contrast, in the disordered (E)-benzyl\u00adidene\u00adamino substituent, the corresponding torsion angles N2\u2014C5\u2014C21\u2014C22 and N2\u2032\u2014C5\u2032\u2014C21\u2032\u2014C22\u2032 are \u221217.6\u2005(6) and 21\u2005(4)\u00b0 for the major and minor disorder components, respectively. All these data suggest that the difference between these (E)-benzyl\u00adidene\u00adamino substituents may result from some loss of conjugation between the phenyl ring and its azomethine substituent in the disordered branch of the mol\u00adecule.The N1=C1 and N2=C5 distances in the mol\u00adecule are 1.270\u2005(2) and 1.259\u2005(3)\u2005\u00c5, respectively, consistent with C=N double bonding. The C1\u2014N1\u2014C2 bond angle of 118.61\u2005(15)\u00b0 confirms the C(5) chains along the b-axis direction. In addition, pairs of the chains are linked by weak C24\u2014H24\u22efCg1 inter\u00adactions bis\u00ad] bis\u00ad(4-methyl\u00adbenzene\u00adsulfonamide) -(2-chloro\u00adbenzyl\u00adidene)amino]\u00adpropan-2-ol -(4-meth\u00adoxy\u00adbenzyl\u00adidene)amino]\u00adpropan-2-ol = 0.95\u2005\u00c5 for aromatic and azomethine atoms, d(C\u2014H) = 0.99\u2005\u00c5 for methyl\u00adene and d(C\u2014H) = 1.00\u2005\u00c5 for C3\u2014H3. The Uiso(H) values were constrained to 1.2Ueq(C). The C3/C4/N2/C5/C21\u2013C26 segment of the mol\u00adecule is disordered over two sets of sites with a refined occupancy ratio of 0.851\u2005(4):0.149\u2005(4).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017004741/sj5523sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017004741/sj5523Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017004741/sj5523Isup3.cmlSupporting information file. DOI: 1540296CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Mecp2ZFN\u0394/y rats display respiratory abnormalities early in development.Post-publication, the authors discovered a scale factor error of 5 affecting volumetric readings from the whole-body plethysmography chamber used for juvenile male rats (PND 21-28) in this study (Buxco PLY3211). As such, reported tidal volume (TV) and minute ventilation (MV) data are erroneously high for this group. This change does not affect the interpretation of this data as the scale factor change is applicable to both groups being compared. The below figure shows the corrected graphical representation of juvenile male rats' TV and MV readings . All numerical values have been reduced by a factor of 5 to correct for the recognized instrument error. The text of the results section has been updated online to reflect this numerical correction and should be as follows:Mecp2ZFN\u0394/y rats exhibited abnormal breathing compared to WTs . Juvenile WTs breathed at a rate of 126.6\u2009\u00b1\u20095.3 BPM, which is comparable to respiratory rates of similarly aged Sprague-Dawley rats (46); however, age-matched Mecp2ZFN\u0394/y rats breathed at a rate of only 100.1\u2009\u00b1\u20094.7 BPM . The reduction in frequency was due to both an increase in inspiratory time (Ti) (0.20\u2009\u00b1\u20090.01\u2009s WT vs. 0.25\u2009\u00b1\u20090.01\u2009s Mecp2ZFN\u0394/y) and expiratory time (Te) (0.28\u2009\u00b1\u20090.01\u2009s WT vs. 0.37\u2009\u00b1\u20090.03\u2009s Mecp2ZFN\u0394/y) . Despite these changes in respiratory frequency, juvenile Mecp2ZFN\u0394/y males exhibit an otherwise normal tidal volume (9.1\u2009\u00b1\u2009.6\u2009ml/kg WT vs. 10.6\u2009\u00b1\u2009.6\u2009ml/kg Mecp2ZFN\u0394/y) , minute ventilation (1151.6\u2009\u00b1\u200985.0\u2009ml/min/kg WT vs. 1073.7\u2009\u00b1\u2009101.1\u2009ml/min/kg Mecp2ZFN\u0394/y) , a relatively stable respiratory cycle (irregularity score\u2009=\u20095.0\u2009\u00b1\u20090.6\u2009WT vs. 5.1\u2009\u00b1\u20090.6 Mecp2ZFN\u0394/y) , and a similar number of spontaneous apneas , but of a shorter duration compared to controls .Respiratory activity was measured in juvenile male (PND 21-28), adult male (PND 40+), and aged (18 month) female rats by whole-body plethysmography. We found that under control conditions (awake and breathing room air) juvenile"} +{"text": "In the mol\u00adecule, the hydroxyl substituent is bound intra\u00admolecularly to the aldehyde group at the ortho position.Wavy layers of mol\u00adecules are formed in the crystal structure of 5-methyl\u00adsalicyl\u00adaldehyde due to weak C\u2014H\u22efO inter\u00adactions between methyl groups and the aromatic ring system. Mol\u00adecules form columns in which the methyl groups are oriented in opposite directions layer-by-layer along cell axis 5-MSA; systematic name 2-hy\u00addroxy-5-methyl\u00adbenzaldehyde), C8H8O2, was discovered to be a textbook example of the drastic structural changes caused by just a few weak C\u2014H\u22efO inter\u00adactions due to the additional methyl\u00adation of the aromatic ring compared to salicyl\u00adaldehyde SA. This weak inter\u00admolecular hydrogen bonding is observed between aromatic or methyl carbon donor atoms and hydroxyl or aldehyde acceptor oxygen atoms with d(D\u22efA) = 3.4801\u2005(18) and 3.499\u2005(11)\u2005\u00c5. The mol\u00adecule shows a distorted geometry of the aromatic ring with elongated bonds in the vicinity of substituted aldehyde and hydroxyl carbon atoms. The methyl hydrogen atoms are disordered over two sets of sites with occupancies of 0.69\u2005(2) and 0.31\u2005(2).The crystal structure of 5-methyl\u00adsalicyl\u00adaldehyde ( Its crystal structure is reported herein and compared to the unsubstituted form of salicyl\u00adaldehyde (SA) apart from van der Waals inter\u00adactions. Three C\u2014H\u22efO inter\u00adactions are present between either aromatic or methyl C atoms and aldehyde or alcohol oxygen atoms: two close to 3.5\u2005\u00c5 with C10\u22efO8 = 3.499\u2005(2)\u2005\u00c5 and C5\u22efO8 = 3.4801\u2005(18)\u2005\u00c5 and corresponding C\u2014H\u22efO angles of 152 and 149.3\u2005(13)\u00b0, respectively. The third and shortest inter\u00adaction, has a C6\u22efO9 distance of 3.4053\u2005(18)\u2005\u00c5 and an angle of 138.7\u2005(12)\u00b0 = 3.7539\u2005(11) and 4.7456\u2005(13)\u2005\u00c5, respectively. This results in a deviation from the usually expected herringbone or completely planar arrangement of planar mol\u00adecules. Wavy layers of mol\u00adecules are formed instead, whereby the 5-MSA mol\u00adecules form columns in which the methyl groups are oriented in opposite directions layer-by-layer along the a axis at positions H10A, H10B, H10C and 0.31\u2005(2) at positions H10D, H10E, H10F. They were included at idealized positions riding on the parent carbon atom, with isotropic displacement parameters iUso(H) = 1.5Ueq(CH3). Refinement of the corresponding site-occupation factors of the methyl-group hydrogen atoms was carried out using a free variable so that their sum is unity. All other hydrogen atoms were located individually in a difference-Fourier map and refined isotropically.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017000238/lh5831sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017000238/lh5831Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017000238/lh5831Isup3.cmlSupporting information file. DOI: 1525796CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecule has an {Na2O6(\u03bc-O)2} core with two bridging carbonyl O atoms and two hydroxamate O atoms of two mono-deprotonated residues of N-hy\u00addroxy\u00adpicolinamide, while two neutral N-hy\u00addroxy\u00adpicolinamide mol\u00adecules are coordinated in a monodentate manner to each sodium ion via the carbonyl O atoms [the Na\u2014O distances range from 2.3044\u2005(2) to 2.3716\u2005(2)\u2005\u00c5]. The penta\u00adcoordinated sodium ion exhibits a distorted trigonal\u2013pyramidal coordination polyhedron. In the crystal, the coordination dimers are linked into chains along the c axis via N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds; the chains are linked into a two-dimensional framework parallel to (100) via weak C\u2014H\u22efO and \u03c0\u2013\u03c0 stacking inter\u00adactions.The title compound, [Na Coordination of the \u03bc-O carbonyl and hydroxamate O atoms of the same anion lead to the formation of five-membered chelate rings [Na1\u2014O6i = 2.3716\u2005(14)\u2005\u00c5 and O5i\u2014Na\u2014O6i = 70.26\u2005(5)\u00b0]. Two neutral N-hy\u00addroxy\u00adpicolinamide mol\u00adecules coordinate in a monodentate manner to each sodium ion via the carbonyl O atoms [Na1\u2014O1 = 2.3300\u2005(16)\u2005\u00c5 and Na1\u2014O3 = 2.3225\u2005(15)\u2005\u00c5]. As a result, each penta\u00adcoordinated sodium ion reveals a distorted trigonal\u2013pyramidal coordination polyhedron \u2005\u00c5 and the deviation of the O\u2014Na\u2014O angles from ideal values are up to 23.47\u2005(5)\u00b0. The Na\u2014O bond lengths are in the range 2.3044\u2005(14)\u20142.3716\u2005(14)\u2005\u00c5, which is common for penta\u00adcoordinated sodium cations 2 core is virtually planar and approaches a square [the O\u2014Na\u2014O angles are 86.43\u2005(5) and 93.57\u2005(5)\u00b0].The mol\u00adecular structure of title compound is shown in Fig.\u00a01N-hy\u00addroxy\u00adpicolinamide act as donors . The nearly coplanar pyridine rings of two neutral mol\u00adecules of N-hy\u00addroxy\u00adpicolinamide coordinating to the same sodium ion reveal intra\u00admolecular stacking inter\u00adactions in unusual \u2018head-to-head\u2019 manner .The deprotonated hydroxamate atom O6 acts as an acceptor of two hydrogen bonds Table\u00a01 in whichN-hy\u00addroxy\u00adpicolinamide residue adopts a strongly flattened conformation with a dihedral angle of only 0.6\u2005(2)\u00b0 between the hydroxamic group and the pyridine ring. At the same time, the corresponding dihedral angles in both neutral N-hy\u00addroxy\u00adpicolinamide mol\u00adecules are noticeably greater [17.5\u2005(2) and 8.9\u2005(2)\u00b0], indicating a deviation of the hydroxamic group from the plane of pyridine rings. The configuration about the hydroxamic C\u2014N bond is Z and that about the C\u2014C bond between the pyridine and hydroxamic groups is E for both the neutral and deprotonated hydroxamates. Intra\u00admolecular N\u2014H\u22efN attractive contacts between the hydroxamate group and the nitro\u00adgen atom of pyridine ring [2.25\u2005(2)\u20132.35\u2005(3)\u2005\u00c5] are present in both the neutral and deprotonated N-hy\u00addroxy\u00adpicolinamide mol\u00adecules and N1\u2014H1\u22efN6 hydrogen bonds supported by a pair of weak non-classical C17\u2014H17\u22efN2 hydrogen bonds hydrogen bond and \u03c0\u2013\u03c0 stacking between the N4/C7\u2013C11 pyridine ring and the deprotonated O5/C18/N5/O6 hydroxamic group . Inter\u00admolecular \u03c0\u2013\u03c0 stacking between the same deprotonated hydroxamic group and the N2/C1\u2013C5 pyridine ring links the frameworks into a three-dimensional structure.In the crystal Fig.\u00a02, the dims Table\u00a01. The chaet al., 2016N-hy\u00addroxy\u00adpicolinamide revealed the crystal structures of over 20 compounds, mostly belonging to the metallacrown (MC) family. In particular, heterometallic copper(II) 15-metallacrown-5 complexes with encapsulated GdIII and EuIII ions ,picHA-4](OTf)1.25(OH)0.75 ,picHA-4]2(OH)3(py)2 ,picHA-4]2\u00b7\u00b7(pyridine)8\u00b7(triflate)3 2(OAc)4(DMF)2 collapsed metallacrown complex has been struct\u00adurally characterized with sodium hydrogen carbonate . Colorless crystals suitable for X-ray diffraction were obtained from the resulting aqueous solution by slow evaporation at ambient temperature within 48\u2005h (yield 78%).The title compound was obtained by the reaction of Uiso = 1.2Ueq(C), and H atoms of the N\u2014H and O\u2014H groups were refined isotropically.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016019095/xu5895sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016019095/xu5895Isup2.hklStructure factors: contains datablock(s) I. DOI: 1520114CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The HCO3\u2212 transport across the luminal membrane has been proposed to be mediated by SLC26A Cl\u2212\u2013HCO3\u2212 exchangers. To examine the electrophysiological properties of Cl\u2212\u2013HCO3\u2212 exchangers, we directly measured HCO3\u2212 conductance in the luminal membrane of the interlobular pancreatic duct cells from guinea pigs using an inside-out patch-clamp technique. Intracellular HCO3\u2212 increased the HCO3\u2212 conductance with a half-maximal effective concentration value of approximately 30\u00a0mM. The selectivity sequence based on permeability ratios was SCN\u2212 (1.4)\u2009>\u2009Cl\u2212 (1.2)\u2009=\u2009gluconate (1.1)\u2009=\u2009I\u2212 (1.1)\u2009=\u2009HCO3\u2212 (1.0)\u2009>\u2009methanesulfonate (0.6). The sequence of the relative conductance was HCO3\u2212 (1.0)\u2009>\u2009SCN\u2212 (0.7)\u2009=\u2009I\u2212 (0.7)\u2009>\u2009Cl\u2212 (0.5)\u2009=\u2009gluconate (0.4)\u2009>\u2009methanesulfonate (0.2). The current dependent on intracellular HCO3\u2212 was reduced by replacement of extracellular Cl\u2212 with gluconate or by H2DIDS, an inhibitor of Cl\u2212\u2013HCO3\u2212 exchangers. RT-PCR analysis revealed that the interlobular and main ducts expressed all SLC26A family members except Slc26a5 and Slc26a8. SLC26A1, SLC26A4, SLC26A6, and SLC26A10 were found to be localized to the luminal membrane of the guinea pig pancreatic duct by immunohistochemistry. These results demonstrate that these SLC26A Cl\u2212\u2013HCO3\u2212 exchangers may mediate the electrogenic HCO3\u2212 transport through the luminal membrane and may be involved in pancreatic secretion in guinea pig ducts.The pancreatic duct epithelium secretes the HCO However, ducts secrete a HCO3\u2212-rich fluid, which is dependent on the presence of CO2/HCO3\u2212-buffer, and that neutralizes acid chyme in the duodenum [3\u2212 transport involves Cl\u2212\u2013HCO3\u2212 exchangers that operate in parallel with cAMP-activated Cl\u2212 channels [cystic fibrosis transmembrane conductance regulator (CFTR)] and Ca2+-activated Cl\u2212 channels, such as TMEM16A/ANO1, on the luminal membranes of duct cells -5-thiazolidinylidene]methyl]benzoic acid and 2-methyl-8-(phenylmethoxy)imidazopyridine-3-acetonitrile were dissolved in DMSO at a 1000-fold concentration for application. The current was recorded in the inside-out configuration using the EPC 800 patch-clamp amplifier (HEKA). The amplifier was driven by Clampex 9 (Axon) in order to allow the delivery of a voltage-ramp protocol with concomitant digitization of the current. The membrane potential was generally held at 0\u00a0mV, and the command voltage was varied from \u2212\u200980 to +\u200980\u00a0mV over a duration of 800\u00a0ms every 10\u00a0s.Standard patch-clamp techniques were used. Patch pipettes, pulled from capillaries of hard borosilicate glass , had a resistance of 5\u20137\u00a0M\u03a9 when filled with a standard Taq DNA Polymerase (Promega). The amplification parameters used were as follows: 1 cycle at 50\u00a0\u00b0C for 30\u00a0min and 1 cycle at 95\u00a0\u00b0C for 15\u00a0min, followed by 40\u00a0cycles at 94\u00a0\u00b0C for 30\u00a0s, 57\u00a0\u00b0C for 30\u00a0s, 72\u00a0\u00b0C for 30\u00a0s, and 1 cycle at 72\u00a0\u00b0C for 10\u00a0min. The transcripts were subsequently verified by agarose gel electrophoresis.RNA was extracted from the interlobular (outside diameter of 50\u2013150\u00a0\u03bcm) and main ducts (outside diameter of around 500\u00a0\u03bcm) from three independent guinea pigs using the RNeasy Plus Micro kit with DNase I (Qiagen). RT-PCR analysis was performed using the OneStep RT-PCR kit (Qiagen) with primers designed to recognize different types of transporters (Table n\u2009=\u20093) in accordance with protocols approved by the Animal Experimentation Committee, Kansai Medical University. The guinea pigs were anesthetized with isoflurane and a mixture of medetomidine (0.5\u00a0mg/kg body weight), midazolam (5.0\u00a0mg/kg b.w.), and butorphanol (2.5\u00a0mg/kg b.w.), and perfused transcardially with 4% paraformaldehyde. The pancreas was fixed with 4% paraformaldehyde in PBS for 24\u00a0h, embedded in paraffin, and sectioned. Detailed methods for immunohistochemistry were described previously [Immunolocalization was performed on the guinea pig pancreas. The pancreas was obtained from female Hartley guinea pigs on ice for 30\u00a0min, and then centrifuged. The pellet was solubilized in lysis buffer containing urea (9\u00a0M), Triton X-100 2%), dithiothreitol (1%), and lithium dodecyl sulfate (2%). The samples (30\u00a0\u03bcg/lane protein) were fractionated on SDS polyacrylamide gel (7.5%), electroblotted to PVDF membranes (Merck Millipore), blocked with skim milk (1%), and reacted with anti-SLC26A4, anti-SLC26A6, or anti-SLC26A10 antibodies or Student\u2019s paired I\u2013V) relationships in the presence of intracellular HCO3\u2212 at different concentrations . As we used the standard NMDG-Cl pipette solution and the bathing solution containing KHCO3, the inward current was due to HCO3\u2212 efflux through the luminal membrane. The inward conductance determined from the linear section of the I\u2013V relationships (from \u2212\u200980 to \u2212\u200960\u00a0mV) increased with intracellular HCO3\u2212 concentration value for the effects of HCO3\u2212 and Hill coefficient were 31.5\u2009\u00b1\u20095.1\u00a0mM and 3.5\u2009\u00b1\u20090.4 (n\u2009=\u20095), respectively. We also measured inward HCO3\u2212 currents in the bathing solution containing 130\u00a0mM NaHCO3. The inward conductance increased to 2.04\u2009\u00b1\u20090.95\u00a0nS in NaHCO3 from 0.34\u2009\u00b1\u20090.08\u00a0nS in NaCl . Thus, there was a minor contribution of K+ conductance under unstimulated conditions.We recorded macroscopic currents from excised inside-out patches from the luminal membrane of the interlobular pancreatic duct cells of guinea pigs under unstimulated conditions. Figure 3\u2212 in the intracellular bathing solution with other monovalent anions. Figure I\u2013V relations recorded in the inside-out configuration with the standard NMDG-Cl pipette solution. In experiments where HCO3\u2212 in the bath was replaced with Cl\u2212 or gluconate (glc\u2212), the reversal potential did not change, but inward conductance significantly decreased from 1.30\u2009\u00b1\u20090.09\u00a0nS in HCO3\u2212 to 0.64\u2009\u00b1\u20090.13\u00a0nS in Cl\u2212 and from 1.69\u2009\u00b1\u20090.08\u00a0nS in HCO3\u2212 to 0.72\u2009\u00b1\u20090.14\u00a0nS in glc\u2212 . Replacement of HCO3\u2212 with methanesulfonate (MES\u2212) shifted the reversal potential in a negative direction, indicating it was less permeable than HCO3\u2212, and the inward conductance significantly decreased from 2.56\u2009\u00b1\u20090.73\u00a0nS in HCO3\u2212 to 0.73\u2009\u00b1\u20090.36\u00a0nS in MES\u2212 . Replacement of HCO3\u2212 with thiocyanate (SCN\u2212) slightly shifted the reversal potential in a positive direction, but the inward conductance had little change . Finally, replacement of HCO3\u2212 with iodide (I\u2212) did not cause a marked difference in the reversal potential or the inward conductance . We calculated the permeability ratio (PX/PHCO3) from the shift in the reversal potential (\u0394Vrev) when anion X is substituted for internal HCO3\u2212 [\u2212 (1.41\u2009\u00b1\u20090.15)\u2009>\u2009Cl\u2212 (1.18\u2009\u00b1\u20090.14)\u2009=\u2009glc\u2212 (1.07\u2009\u00b1\u20090.03)\u2009=\u2009I\u2212 (1.06\u2009\u00b1\u20090.06)\u2009=\u2009HCO3\u2212 (1.00)\u2009>\u2009MES\u2212 (0.65\u2009\u00b1\u20090.11) (n\u2009=\u20095\u20136). The sequence of the relative inward conductance determined from \u2212\u200980 to \u2212\u200960\u00a0mV was HCO3\u2212 (1.00)\u2009>\u2009SCN\u2212 (0.69\u2009\u00b1\u20090.10)\u2009=\u2009I\u2212 (0.66\u2009\u00b1\u20090.09)\u2009>\u2009Cl\u2212 (0.48\u2009\u00b1\u20090.08)\u2009=\u2009glc\u2212 (0.43\u2009\u00b1\u20090.09)\u2009>\u2009MES\u2212 (0.26\u2009\u00b1\u20090.06) (n\u2009=\u20095\u20136).Ion selectivity of the bicarbonate conductance was examined by replacing 130\u00a0mM HCO\u2212\u2013HCO3\u2212 exchangers on the apical membrane of interlobular pancreatic ducts of the guinea pig, Ishiguro and colleagues replaced Cl\u2212 with gluconate in the lumen [\u2212. With the control intracellular solution,Erev was \u2212\u200946.6\u2009\u00b1\u20094.5\u00a0mV with a standard NMDG-Cl pipette solution and \u2212\u200963.6\u2009\u00b1\u20093.9\u00a0mV with gluconate-rich pipette solution , demonstrating a significant difference (ANOVA). We also compared the inward HCO3\u2212 conductance with gluconate-rich and standard NMDG-Cl pipette solutions than with standard NMDG-Cl pipette solutions (1.30\u2009\u00b1\u20090.09\u00a0nS) . Additionally, as described in the previous section, the inward conductance significantly decreased when HCO3\u2212 in the bath was replaced with Cl\u2212, indicating that there was a minor contribution from Cl\u2212-dependent current intracellularly while recording macroscopic currents from excised inside-out patches with the control bicarbonate internal and the standard NMDG-Cl pipette solutions. To exclude the possibility of the contamination of CFTR Cl\u2212 conductance, we included 20\u00a0\u03bcM CFTRinh-172, an inhibitor of CFTR Cl\u2212 channels, in the pipette solution. H2DIDS applied intracellularly significantly decreased inward HCO3\u2212 conductance from 1.57\u2009\u00b1\u20090.55 to 0.86\u2009\u00b1\u20090.37\u00a0nS . We also tested 30\u00a0\u03bcM Sch28080, a H+\u2013K+-pump inhibitor, but did not observe any inhibitory effects on the inward HCO3\u2212 conductance . These results further support that the HCO3\u2212 conductance occurs through Cl\u2212\u2013HCO3\u2212 exchangers.Previous studies reported that Clstilbene , 43. For2+ signaling pathways play a role in fluid secretion. As CFTR Cl\u2212 channels were regulated by intracellular cAMP [n\u2009=\u200913). The addition of 1\u00a0mM cAMP further increased the inward conductance to 14.8\u2009\u00b1\u20095.57\u00a0nS (n\u2009=\u20094). cAMP also activated the marked outward conductance, which was attributed to Cl\u2212 influx, most likely through CFTR Cl\u2212 channels. Therefore, we tested the effects of intracellular ATP-Mg and cAMP with the pipette solution including CFTRinh-172 at 20\u00a0\u03bcM. In the presence of CFTRinh-172, application of intracellular 2\u00a0mM ATP-Mg and 1\u00a0mM cAMP had little effect on the conductance in either direction . These results indicate that intracellular ATP and cAMP may not directly regulate Cl\u2212\u2013HCO3\u2212 exchangers, but instead regulate CFTR Cl\u2212 channels on the luminal membrane of duct cells. Additionally, 1\u00a0\u03bcM free Ca2+ added to the intracellular solution did not affect the inward HCO3\u2212 conductance , suggesting that intracellular Ca2+ does not regulate Cl\u2212\u2013HCO3\u2212 exchangers directly.In pancreatic duct cells, cAMP and Calar cAMP , 8, 29 alar cAMP , we test\u2212\u2013HCO3\u2212 exchangers [Slc26a3 and Slc26a6 [n\u2009=\u20093 animals) and main ducts , along with GAPDH and a duct marker of carbonic anhydrase II (CA2). We also screened all primer sets from the total RNA of the kidney as a positive control . The molecular mass values corresponded to those of human SLC26A proteins (~\u2009100\u00a0kDa), which were N-glycosylated, expressed in HEK-293 cells [We next performed western blot analysis to examine the expression of SLC26A protein in the guinea pig pancreatic ducts. We detected SLC26A6 (~\u2009107\u00a0kDa), SLC26A1 ~\u200978\u00a0kDa), SLC26A4 (~\u2009136\u00a0kDa), and SLC26A10 (~\u2009108\u00a0kDa) in the lysates of the isolated ducts of 160 and 80\u00a0\u03bcM, respectively [\u2212\u2013HCO3\u2212 exchangers from not only the outside but also from the inside of the cell membrane.We found that Hane Fig. . A previectively . It is l3\u2212 conductance through the luminal membrane is mediated by Cl\u2212\u2013HCO3\u2212 exchangers under physiological HCO3\u2212 concentrations in pancreatic duct cells. Our findings suggest that SLC26A1, SLC26A4, SLC26A6, and SLC26A10 may be involved in the HCO3\u2212 transport through the luminal membrane. The SLC26A family may also play a role in pH homeostasis in the pancreatic lumen and duct cells. The direct measurement of the HCO3\u2212 current from the interlobular duct and its functional characterization helps to propose a useful model for HCO3\u2212 secretion from the pancreatic duct epithelia (Fig.\u00a0In conclusion, we used the patch-clamp technique in the inside-out configuration and demonstrated that the HCOlia Fig.\u00a0.Fig. 9Mo"} +{"text": "II cations are bridged by S atoms from the N,N-di\u00adallyl\u00adldi\u00adthio\u00adcarbamate ligands.The characteristic feature of this cadmium(II) complex is the formation of a dimeric bridged structure where the two Cd 2(C7H10NS2)4], is a neutral dinuclear cadmium(II) complex bearing four bis N,N-di\u00adallyl\u00addi\u00adthio\u00adcarbamate ligands coordinating to two CdII cations. In each of the monomeric subunits, there are four S atoms of two di\u00adthio\u00adcarbamate ligands that coordinate to one CdII atom in a bidentate mode. The dimers are located over an inversion centre bridged by two additional bridging Cd\u2014S bonds [2.6021\u2005(3)\u2005\u00c5], leading to a substantial distortion of the geometry of the monomeric subunit from the expected square-planar geometry. The five-coordinate environment around each of the CdII ions in the dimer is best described as substanti\u00adally tetra\u00adgonally distorted square pyramidal. The di\u00adthio\u00adcarbamate groups are themselves planar and are also coplanar with the CdII ions. The negative charge on these groups is delocalized by resonance across the S atoms bound to the CdII cation. This delocalization of the \u03c0 electrons in the di\u00adthio\u00adcarbamate groups also extends to the C\u2014N bonds as they reveal significant double bond character [C\u2014N = 1.3213\u2005(16) and 1.3333\u2005(15)\u2005\u00c5].The title compound, [Cd This common feature has been ascribed to the effect of aggregated species, which they adopt in the solid state, resulting from equal numbers of \u03bc2-tridentate and bidentate (chelating) ligands cadmium compounds have the advantage of having stability similar to that of the zinc complexes, but more favourable stability when compared to the mercury complexes. Cadmium di\u00adthio\u00adcarbamate complexes have been widely used as single-source precursors for CdS nanoparticles and thin films, which have application as non-linear optical materials to 2.8016\u2005(3)\u2005\u00c5 and to a fifth S atom at a distance of 2.6021\u2005(3)\u2005\u00c5; these distances are similar to other complexes found to have been published previously (see Section 4: Database survey). A full geometry check carried out with the Mogul Geometry Check tool \u2013x\u00a0+\u00a02, \u2013y, \u2013z\u00a0+\u00a01]. This means that each bridging S atom simultaneously occupies an equatorial coordination site on one CdII ion and an apical site on the other CdII ion to form an edge-shared tetra\u00adgonal\u2013pyramidal geometry. The CdII ion deviates from the S11\u2014S12\u2014S22\u2014S21 mean plane by 0.704016\u2005(17)\u2005\u00c5 towards S12i. The bridging network Cd1\u2014S12\u2014Cd1i\u2014S12i is completely planar since it lies over the inversion centre with a Cd1\u22efCd1i separation distance of 3.60987\u2005(8)\u2005\u00c5 and S12\u2014Cd1\u2014S12i and Cd1\u2014S12\u2014Cd1i angles of 96.257\u2005(9) and 83.743\u2005(9)\u00b0, respectively. There is substantial distortion of the geometry of the monomeric subunit from the expected square-planar geometry. Deviations from the standard 90\u00b0 angles are evident in the angles of S11\u2014Cd1\u2014S21 [108.203\u2005(11)\u00b0]; S22\u2014Cd1\u2014S21 [70.264\u2005(10)\u00b0]; S22\u2014Cd1\u2014S12 [96.950\u2005(10)\u00b0] and S11\u2014Cd1\u2014S12 [67.486\u2005(10)\u00b0]. Deviations in the standard 180\u00b0 angles are evident in the angles of S11\u2014Cd1\u2014S22 [143.705\u2005(13)\u00b0] and S21\u2014Cd1\u2014S12 [152.651\u2005(11)\u00b0]. The Cd1\u2014S12\u2014Cd1i\u2014S12i and S11\u2014S12\u2014S22\u2014S21 mean planes form a dihedral (twist) angle of 84.6228\u2005(18)\u00b0. The di\u00adthio\u00adcarbamate groups are planar and each group of the monomeric subunit is coplanar with the CdII ion (r.m.s. deviation is 0.010\u2005\u00c5). The mean plane consisting of atoms Cd1, S11, N1, C11, S12 and the mean plane consisting of atoms Cd1, S22, N2, C21, S21 have a plane-normal-to-plane-normal angle of 37.0291\u2005(10)\u00b0; a centroid-to-centroid distance of 4.45354\u2005(8)\u2005\u00c5; a plane-to-plane shift of 4.22298\u2005(8)\u2005\u00c5 and a plane-to-plane torsion (twist) angle of 8.0304\u2005(12)\u00b0.The coordination environment of the Cdry Fig.\u00a01. The CdIII cation. On the opposite side of the CdII ion, both S\u2014C bonds have approximately the same length, where S21\u2014C21 and S22\u2014C21 bond lengths are 1.7224\u2005(12) and 1.7263\u2005(12)\u2005\u00c5, respectively, suggesting that the double bond of the di\u00adthio\u00adcarbamate is spread over the S\u2014C\u2014S bond via resonance. A possible explanation for this may be because of the fact that atom S12 serves as the bridging S atom in the complex. Also, the N1\u2014C11 and N2\u2014C21 distances are shorter compared to the other N\u2014C distances indicating considerable double-bond character. The vinyl substituents are also planar and are at an angle of 91.6049\u2005(14)\u00b0 from the di\u00adthio\u00adcarbamate plane and at an angle of 150.9196\u2005(6)\u00b0 from the vinyl group directly opposite from it. This scenario is comparable with the other structures surveyed in the literature (see Section 4: Database survey). All highlighted and discussed geometrical parameters describing the coordination environment are given in Table\u00a01The S12\u2014C11 bond length [1.7532\u2005(13)\u2005\u00c5] is longer than the adjacent S11\u2014C11 bond length [1.7162\u2005(13)\u2005\u00c5] suggesting that this bond has more double bond character in the di\u00adthio\u00adcarbamate portion that coordinates to the CdPThe space group of the crystal is et al., 2016N,N-diallyl side chain, the side-chains substituents are di-n-propyl [CSD refodes BEHNOR in ethanol (10\u2005ml) was added to a solution of sodium N,N-diallyl di\u00adthio\u00adcarbamate in ethanol (10\u2005ml), and the resulting suspension was stirred for 45\u2005min at room temperature. This solution was then filtered, and rinsed several times with distilled water has been published previously = 1.2 Ueq(C) for methyl\u00adene groups and C\u2014H = 0.95\u2005\u00c5 and Uiso(H) = 1.2 Ueq(C) for all vinyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989017011616/zl2710sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017011616/zl2710Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017011616/zl2710Isup3.molSupporting information file. DOI: 899314CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N\u2032-amino\u00adpyridine-2-carboximidamide (C6H8N4), 1, and N\u2032-{[1-(pyridin-2-yl)ethyl\u00adidene]amino}\u00adpyridine-2-carboximidamide (C13H13N5), 2, mol\u00adecules are linked by inter\u00admolecular N\u2014H\u22efN hydrogen-bonding inter\u00adactions, forming a two-dimensional network in 1 and a chain in 2.In the crystal structures of N\u2032-amino\u00adpyridine-2-carboximidamide (C6H8N4), 1, and N\u2032-{[1-(pyridin-2-yl)ethyl\u00adidene]amino}\u00adpyridine-2-carboximidamide (C13H13N5), 2, are described. The non-H atoms in compound 1 are nearly planar (r.m.s. deviation from planarity = 0.0108\u2005\u00c5), while 2 is twisted about the central N\u2014N bond by 17.8\u2005(2)\u00b0. Both mol\u00adecules are linked by inter\u00admolecular N\u2014H\u22efN hydrogen-bonding inter\u00adactions; 1 forms a two-dimensional hydrogen-bonding network and for 2 the network is a one-dimensional chain. The bond lengths of these mol\u00adecules are similar to those in other literature reports of azine and di\u00adimine systems.The crystal structures of RC(=NH)NHNH2 is accomplished by the action of hydrazine on the corresponding thio\u00adamide, imido ether or nitrile of the heterocyclic ligand and 0.24\u2005(6)\u2005\u00c5 out of the mean plane of non-hydrogen atoms. For H4A and H4B, the deviation is even greater at 0.37\u2005(5) and 0.54\u2005(5)\u2005\u00c5 from the mean plane. Rotation of the non-planar NH2 group, particularly for N4, facilitates hydrogen bonding to other mol\u00adecules. The N\u2014N single bond length in 1 [1.424\u2005(5)\u2005\u00c5] is slightly shorter than that in the free hydrazine (1.449\u2005\u00c5).The mol\u00adecular structure of 2 is shown in Fig.\u00a02The mol\u00adecular structure of E,E conformation, suggesting conjugation throughout the \u03c0 systems. The C6\u2014N2\u2014N3 and C8\u2014N3\u2014N2 angles of 115.5\u2005(2)\u00b0 and 110.57\u2005(19)\u00b0, respectively are significantly below the ideal sp2 value of 120\u00b0, a consequence of the repulsion between the nitro\u00adgen lone pair and the adjacent bonds. The C6\u2014N2\u2014N3\u2014C8 torsion angle is \u2212162.2\u2005(2)\u00b0. This large deviation from planarity has two consequences. First, there is a loss of conjugation between the imine bonds across the azine bond, reflected in the shorter imine bond length. The torsion also leads to a shorter N2\u2014N3 bond length [1.408\u2005(3)\u2005\u00c5] compared to that observed for 1 [1.424\u2005(5)\u2005\u00c5]. Finally, a short intra\u00admolecular contact between N3i and H4B, 2.42\u2005(3)\u2005\u00c5, may add a favorable electrostatic contribution to the stability of this conformation. Notably, there is minimal change in the bond lengths within the ligands when a first row transition metal ion is bound. When the ligand chelates to a metal ion through both N3 and N5, only the bond length C8\u2014N4 changes significantly, becoming shorter on binding.The bond lengths indicate that within the central chain of the mol\u00adecule, the C6\u2014N2 and C8\u2014N3 linkages have largely double-bond character. The azine linkages are in the 1 in each unit cell and these are related by the screw axis. Curiously, N1 does not act as a hydrogen-bond acceptor. H2A is also not involved with the formation of any (short) classical hydrogen bonds. H2B forms a hydrogen bond to N4i . This is augmented by the longer hydrogen bond N4\u2014H4B\u22efN4i. N4\u2014H4A forms a hydrogen bond to N3ii . These three sets of hydrogen bonds 1/2 \u2013 x, 1 \u2013 y, z \u2013 1/2] is present and this knits the mol\u00adecules of 2 together to form hydrogen-bonded chains along the [001] direction, as shown in Fig.\u00a04The classical hydrogen bonding Table\u00a02 in 2 is N\u2032-aminopyridine-2-carboximidamide and related mol\u00adecules, see Case et al. (1965N\u2032-{[1-(pyridin-2-yl)ethylidene]amino}pyridine-2-carboximidamide and analogues, see Gokhale et al. ethyl\u00adidene]amino}\u00adpyridine-2-carboximid\u00adamide is depicted in Fig.\u00a05The synthesis of N\u2032-Amino\u00adpyridine-2-carboximidamide (1) was prepared by an analogy of the procedure published by Case (1965N\u2032-{[1-(Pyridin-2-yl)ethyl\u00adidene]amino}\u00adpyridine-2-carboximid\u00adamide (2) was synthesized by an analogy of the procedure published by Gokhale et al. to 0.99\u2005(3)\u2005\u00c5. The other hydrogen atoms attached to formally single-bonded nitro\u00adgen atoms were freely refined subject to sensible distance and angle restraints. The N\u2014H distances lie in the range 0.94\u2005(3)-0.95\u2005(3)\u2005\u00c5.For compound 2, hydrogen atoms were placed using a riding model .For compound 10.1107/S2056989017008416/nk2236sup1.cifCrystal structure: contains datablock(s) 1, 2. DOI: 10.1107/S2056989017008416/nk22361sup2.hklStructure factors: contains datablock(s) 1. DOI: Click here for additional data file.10.1107/S2056989017008416/nk22361sup4.cdxSupporting information file. DOI: 10.1107/S2056989017008416/nk22362sup3.hklStructure factors: contains datablock(s) 2. DOI: Click here for additional data file.10.1107/S2056989017008416/nk22362sup5.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989017008416/nk22361sup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989017008416/nk22362sup7.cmlSupporting information file. DOI: 1554569, 1554568CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Molecules containing multiple bonds between atoms\u2014most often in the form of olefins\u2014are ubiquitous in nature, commerce, and science, and as such have a huge impact on everyday life. Given their prominence, over the last few decades, frequent attempts have been made to perturb the structure and reactivity of multiply-bound species through bending and twisting. However, only modest success has been achieved in the quest to completely twist double bonds in order to homolytically cleave the associated \u03c0 bond. Here, we present the isolation of double-bond-containing species based on boron, as well as their fully twisted diradical congeners, by the incorporation of attached groups with different electronic properties. The compounds comprise a structurally authenticated set of diamagnetic multiply-bound and diradical singly-bound congeners of the same class of compound. Attempts to bend and twist multiple bonds in order to alter their reactivities have thus far been met with only modest success. Here, Braunschweig and colleagues isolate double-bond-containing boron-based species and their 90\u00b0-twisted diradical analogs, thanks to their stabilization with Lewis basic units. Having one degree of electronic unsaturation more than relatively inert alkanes, olefins are the simplest and most earth-abundant hydrocarbons that show reactivity under ambient conditions, leading to their widespread industrial use as well as their implication in questions related to the origins of life4. Olefins are molecules that contain a double bond between two carbon atoms consisting of one bond of \u03c3 symmetry and one of \u03c0 symmetry olefin by 90\u00b0 would completely break its \u03c0 bond, presumably leading to a triplet diradical due to their necessarily twisted geometry.The \u03c0 orbital of olefins requires the parallel alignment of two carbon cal Fig.\u00a0, right a18. Beyond carbon\u2013carbon double bonds, sterically hindered heavier olefin analogs such as disilenes (of the form R2Si\u2009=\u2009SiR2) also withstand significant twisting of the double bond (up to 55\u00b0)23. In a key 2015 study, Sekiguchi and coworkers reported that one of these disilenes (Si2(SiMetBu2)4) was able to thermally populate a triplet state by twisting of the Si=Si bond above 77\u2009\u00b0C. This diradical compound was characterized by electron paramagnetic resonance (EPR) spectroscopy and calculated to have an average SiSiSiSi dihedral angle of 75.2\u00b0; however, its structure could not be authenticated experimentally22. This work remains the forefront of chemists\u2019 efforts to isolate the diradical products of twisting element\u2013element double bonds.It is thus unsurprising that efforts to isolate \u201ctwisted olefin\u201d compounds have resulted in only partial success: isolation of diamagnetic, incompletely twisted structures 27, as well as their 90\u00b0-twisted diradical congeners 2 imidazol-2-ylidene) led to new 11B NMR signals to higher field of that of the triply-bound precursor (\u03b4B 39)24, signifying the formation of three new products, 1a\u2013c or purple solids with 11B NMR signals in a similar range to those of known doubly NHC-stabilized diborenes (\u03b4B 18\u201328)27, but significantly more downfield than those of two previously observed diboratellurirenium cations (cyclo-[Te(R)B(NHC)B(NHC)]+) prepared by the analogous reactions of B2(IDip)2 with diphenylditelluride (\u03b4B 0)35. Single-crystal X-ray diffraction confirmed the trans-diborene nature of 1a\u2013c, all three molecules displaying effectively planar LRB=BRL double bond units \u2009\u00c5; 1b: avg. 1.568(6)\u2009\u00c5; 1c: avg. 1.565(5)\u2009\u00c5).Simple room-temperature (RT) addition of either equal amounts or slight excesses of dibutyldisulfide, diphenyldisulfide, or diphenyldiselenide to solutions of the triple-bond-containing diboryne species Ba\u2013c Fig.\u00a0, top. Afits Fig.\u00a0, top, an2(IDip)2 (B2(CAAC)2; CAAC\u2009=\u20091--3,3,5,5-tetramethylpyrrolidin-2-ylidene)36. However, to our surprise, all three reaction mixtures became very dark, and all attempts to obtain NMR spectra from the mixtures gave either silent spectra or exceptionally broad signals well outside the normal diamagnetic chemical shift ranges. Solvent extraction and recrystallization provided the black solids 2a\u2013c \u00b0; 2b: 100.9(1)\u00b0; 2c: 99.7(2)\u00b0), and long, single B\u2013B bonds (2a: 1.728(2)\u2009\u00c5; 2b: 1.713(2)\u2009\u00c5; 2c: 1.700(4)\u2009\u00c5). On either side of the midpoint of the molecules, the N-C-B-S/Se units are near-planar, with relatively small N-C-B-S/Se torsion angles (2a: 1.7(2)\u00b0, 14.6(2)\u00b0; 2b: 17.4(2)\u00b0, 1.5(2)\u00b0; 2c: 18.7(3)\u00b0, 0.1(4)\u00b0) suggesting significant \u03c0-electron delocalization over this portion of the molecule. The B-S/Se distances of 2a\u2013c (d(B\u2013S): 1.84\u20131.86\u2009\u00c5; d(B\u2013Se): 1.99\u20132.00\u2009\u00c5) were found to be slightly shorter than those of 1a\u2013c (d(B\u2013S): 1.88\u20131.92\u2009\u00c5; d(B\u2013Se): 2.01\u20132.06\u2009\u00c5), and the B\u2013C distances of 2a\u2013c (1.53\u20131.55\u2009\u00c5) are likewise shorter than those of 1a\u2013c (1.57\u20131.62\u2009\u00c5).We also attempted the analogous addition of dibutyldisulfide, diphenyldisulfide, or diphenyldiselenide to solutions of a more cumulenic CAAC analog of Ba\u2013c Fig.\u00a0, bottom.2a\u2013c, the apparent absence of B\u2013B multiple bonding, and the inability to procure NMR data of the compounds suggested that they may be captodatively stabilized30 triplet diradicals, each with two push\u2013pull systems involving the \u03c0-donating S/Se atom and the \u03c0-accepting CAAC carbene carbon atom \u2013B(\u2022)RL; Fig.\u00a02a\u2013c are reminiscent of our related neutral monoradical species [DurB(Cl)(CAAC)]\u2022 reported in 201431. In order to confirm the diradical nature of 2a\u2013c, we performed EPR spectroscopy on 2a\u2013c in frozen 2-methyltetrahydrofuran glass. While we observed an intense half-field transition for 2a and 2b due to dipole interaction, indicating that the spin system in question is a triplet state. Analysis of the spectra yields estimates of the ZFS parameters , the axial parts of which correspond to interspin distances of 4.5 and 4.4\u2009\u00c5 (assuming point dipole approximation), respectively. These distances are significantly longer than the ca. 1.70\u20131.73\u2009\u00c5 boron\u2013boron distances found by X-ray crystallography, and match better the distance between the two CAAC carbene carbon atoms . Thus, the two unpaired electrons must be significantly delocalized into the substituents, most likely toward the carbene carbon atoms, which is in line with that expected by the captodative effect of the \u03c0-donor nitrogen and \u03c0-acceptor boron atoms.The significantly greater delocalization in the C-B-S/Se portion of 2b Fig.\u00a0\u201316, only2a, temperature-dependent magnetic measurements were performed on a solid sample lower than the singlet state. The exchange coupling determined from solid-state SQUID measurements is also reflected to some degree in the temperature-dependence of the signal intensities observed in the solution-state EPR spectra of 2a and single B\u2013B bond character in 2a\u2013c . Figure\u00a01a\u2013c and diradicals 2a\u2013c. The former, as exemplified by 1a, possess highest occupied molecular orbitals , with lower but roughly equivalent spin density on the B (0.18 and 0.23 e) and N (0.21 and 0.22 e) atoms.Density functional theory (DFT) calculations for diamagnetic diborenes 2a Fig.\u00a0 show tha2a. The overestimation may result from the need for multi-reference approaches38 to describe the singlet and triplet states on equal footing. Because 2a is too large for such costly approaches, we investigated the error of the DFT approaches for a smaller model system , the values obtained with DFT range from \u20134 to \u201328\u2009kcal\u2009mol\u20131. The MN12-L/6\u2013311G approach used to estimate the S\u2013T gap of the full system 2a overestimates the stability of the triplet state by about 6\u20137\u2009kcal\u2009mol\u20131. Transferring this error to the predictions of the MN12L41/6\u2013311G43 approach for the full system 2a , a near-zero S\u2013T gap results, which is in line with the measured value.While DFT computations and experiment agree in terms of geometry and the energetic order of the singlet and triplet states, the DFT computations considerably overestimate the size of the singlet\u2013triplet gap (S\u2013T gap) of compound 2a\u2013c also distinguishes these diradicals from diborenes 1a\u2013c , as previously observed for related diborenes44. In contrast, diradicals 2a\u2013c show less-facile oxidation events, with additional reduction waves, in line with the presence of the more \u03c0-accepting CAAC ligands. It should also be noted that variable-temperature NMR experiments suggest that the diamagnetic diborenes 1 and the diradical species 2 do not convert to their alternative conformations upon heating to 80\u2009\u00b0C.The electrochemical behavior of 1a\u2013c and their twisted congeners 2a\u2013c marks the first isolation and structural authentication of diamagnetic, multiply-bound molecules and their diradical, single-bond congeners, products of formal homolysis of the central multiple bond. Crystallographic, EPR spectroscopic, magnetic and theoretical studies confirm that 2a\u2013c are diradical species. Spectroscopic and computational studies also confirm the substantial energy differences between the singlet and triplet states of the molecules, with the singlet states being favored in 1a\u2013c and the triplet states in 2a\u2013c.The synthesis of diborenes 1H: 500.13\u2009MHz, 11B: 160.46\u2009MHz, 13C{1H}: 125.76\u2009MHz) at RT. Chemical shifts (\u03b4) are given in ppm and internally referenced to the carbon nuclei (13C{1H}) or residual protons (1H) of the solvent. 11B NMR spectra were referenced to the external standard [BF3\u00b7OEt2]. Microanalyses were performed on an Elementar vario MICRO cube elemental analyzer. Melting points were determined with a Mettler Toledo 823e DSC in sealed ampules with a ramp rate of 10\u2009\u00b0C\u2009min\u22121. UV/vis absorption spectra 2(SBu)2, (1a): Dibutyldisulfide was added dropwise to a stirred solution of B2(IDip)2 in pentane (10\u2009mL). The mixture was stirred for 18\u2009h at RT and all volatiles were removed in vacuo. The dark green residue was dried in vacuo at 80\u2009\u00b0C for 2\u2009h to remove the excess dibutyldisulfide, extracted with 5\u2009mL pentane, and crystallized from a slowly evaporating hexane/ether solution. The crystalline material was washed with pentane (2\u2009\u00d7\u20091\u2009mL) yielding 73.4\u2009mg of pure 1a as a green solid. 1H NMR : \u03b4\u2009=\u20097.19\u20137.14 , 7.04 , 7.02 , 6.36 , 3.02 , 1.74 , 1.23 , 1.17\u20131.05 , 0.98 , 0.88 . 13C NMR \u03b4\u2009=\u2009176.8 (CNHC), 147.0 (Cq), 139.2 (Cq), 129.1 (CHAryl), 124.4 (CHAryl), 123.8 (CHNHC), 43.1 (SBu-CH2), 34.9 (SBu-CH2), 29.1 (CHiPr), 26.2 (iPr-CH3), 24.3 (iPr-CH3) 24.0 (SBu-CH2), 14.7 (SBu-CH3). 11B NMR : \u03b4\u2009=\u200930 (s) ppm. Elemental analysis calculated for C62H90B2N4S2[+hexane]: C 76.81%, H 9.86%, N 5.27%, S 6.03%; found: C 76.86%, H 9.63%, N 5.62%, S 5.82%.Synthesis of B2(IDip)2(SPh)2, (1b): B2IDip2 and diphenyldisulfide were dissolved in benzene (10\u2009mL) and stirred for 14\u2009h. The solvent was evaporated and the residue was extracted with pentane (2\u2009\u00d7\u20095\u2009mL). The pentane phase was removed under vacuum giving pure 1b as a purple solid. Suitable crystals for X-ray diffraction were obtained by slow evaporation of a pentane solution. Decomp.: 221\u2009\u00b0C. 1H NMR : \u03b4\u2009=\u20097.04 , 6.91 , 6.85 , 6.79 , 6.29 , 3.20 , 1.05 , 0.97 ppm. 13C NMR \u03b4\u2009=\u2009174.7 (Cq), 148.8 (Cq), 146.3 (Cq), 137.6 (Cq), 129.3 (CHAryl), 128.9 (CHAryl), 127.7 (CHAryl), 124.2 (CHAryl), 124.0 (CHNHC), 120.1 (CHAryl), 28.9 (CHiPr), 26.6 (iPr-CH3), 22.7 (iPr-CH3) ppm. 11B NMR : \u03b4\u2009=\u200925 (s) ppm. Elemental analysis calculated for C66H82B2N4S2: C 77.94%, H 8.13%, N 5.51%, S 6.30%; found: C 78.12%, H 8.26%, N 5.58%, S 6.25%.Synthesis of B2(IDip)2(SePh)2, (1c): A solution of B2IDip2 and 1,2-diphenyldiselenide in benzene (10\u2009mL) was stirred for 14\u2009h. After the solvent was removed, the residue was extracted with pentane (2\u2009\u00d7\u20095\u2009mL). The solvent was evaporated under vacuum giving pure 1c as a purple solid. Suitable crystals for X-ray diffraction were obtained by slow evaporation of a pentane solution. Decomp.: 128\u2009\u00b0C. 1H NMR : \u03b4\u2009=\u20097.05 , 6.96 , 6.93 , 6.88 , 6.76 , 6.33 , 3.29 , 1.02 , 0.98 ppm. 13C NMR \u03b4\u2009=\u2009174.7 (Cq) 146.5 (Cq), 143.2 (Cq), 137.53 (Cq), 137.52 (Cq), 131.3 (CHAryl), 129.5 (CHAryl), 127.7 (CHAryl), 124.3 (CHAryl), 124.1 (CHNHC), 121.3 (CHAryl), 29.0 (CHiPr), 26.7 (iPr-CH3), 22.8 (iPr-CH3) ppm. 11B NMR : \u03b4\u2009=\u200922 (s) ppm. 77Se NMR : \u03b4\u2009=\u2009175.6\u2009ppm. Elemental analysis calculated for C66H82B2N4Se2: C 71.36%, H 7.44%, N 5.04%; found: C 70.68%, H 7.66%, N 4.66%.Synthesis of B2(CAAC)2(SBu)2, (2a): Dibutyl disulfide was added dropwise to a stirred solution of B2(CAAC)2 in pentane (20\u2009mL) at \u201340\u2009\u00b0C. The mixture was stirred for 24\u2009h at RT and all volatiles were removed in vacuo. The dark yellow solid was dried in vacuo, dissolved in hexane (10\u2009mL), and black crystals were obtained by slow evaporation of the solvent. The crystalline material was washed with pentane (2\u2009\u00d7\u20091\u2009mL) yielding 291\u2009mg of pure 2a as a black solid. Elemental analysis calculated for C48H80B2N2S2: C 74.78%, H 10.46%, N 3.63%, S 8.32%; found: C 75.15%, H 10.37%, N 3.70%, S 8.01%.Synthesis of B2(CAAC)2(SPh)2, (2b): Pentane (20\u2009mL at \u201340\u2009\u00b0C) was added to a mixture of B2(CAAC)2 and diphenyl disulfide . The mixture was stirred for 3\u2009h at RT, filtered and the residual solid was washed with pentane (3\u2009\u00d7\u20095\u2009mL). The brown solid was dried in vacuo, dissolved in benzene (10\u2009mL), and black crystals were obtained by slow evaporation of the solvent. The crystalline material was washed with pentane (2\u2009\u00d7\u20091\u2009mL) yielding 53.4\u2009mg of pure 2b as a black solid. Decomp.: 138\u2009\u00b0C. Elemental analysis calculated for C52H72B2N2S2: C 77.02%, H 8.95%, N 3.45%, S 7.91%; found: C 77.57%, H 8.97%, N 3.36%, S 7.58%.Synthesis of B2(CAAC)2(SePh)2, (2c): Pentane (20\u2009mL at \u201340\u2009\u00b0C) was added to a mixture of B2(CAAC)2 and diphenyl diselenide . The mixture was stirred for 3\u2009h at RT, filtered and the residual solid was washed with pentane (2\u2009\u00d7\u20095\u2009mL). The brown solid was dried in vacuo, dissolved in benzene (10\u2009mL), and black crystals were obtained by slow evaporation of the solvent. The crystalline material was washed with pentane (2\u2009\u00d7\u20091\u2009mL) yielding 36.2\u2009mg of pure 2c as a black solid. Decomp.: 191\u2009\u00b0C. Elemental analysis calculated for C52H72B2N2Se2: C 69.04%, H 8.02%, N 3.10%; found: C 69.24%, H 7.95%, N 2.74%.45. The EPR spectra of 2a and 2b show an intense half-field signal but only partially resolved zero-field splittings with no visible hyperfine structure. Attempts to improve the resolution by lowering the concentration or changing the solvent were not successful. For 2c, no clear sign of a triplet ground state could be obtained.EPR measurements at the X-band (9.4\u2009GHz) were carried out using a Bruker ELEXSYS E580 CW EPR spectrometer equipped with an Oxford Instruments helium cryostat (ESR900) and a MercuryiTC temperature controller. The spectral simulations were performed using MATLAB 8.6.0.267246 (R2015b) and the EasySpin 5.1.9 toolboxVycor tip, serving as the reference electrode. Formal redox potentials are referenced to the ferrocene/ferrocenium ([Cp2Fe]+/0) redox couple by using ferrocene as an internal standard. Tetrabutylammonium hexafluorophosphate ([Bu4N][PF6]) was employed as the supporting electrolyte. Compensation for resistive losses (iR drop) was employed for all measurements.Cyclic voltammetry experiments were performed using a Gamry Instruments Reference 600 potentiostat. A standard three-electrode cell configuration was employed using a platinum disk working electrode, a platinum wire counter electrode, and a silver wire, separated by a 2IDip2(SBu)2 (1a), B2IDip2(SPh)2 (1b), B2IDip2(SePh)2 (1c), and B2CAAC2(SBu)2 (2a) were collected on a Bruker X8-APEX II diffractometer with a CCD area detector and multi-layer mirror-monochromated MoK\u03b1 radiation. The structure was solved using the intrinsic phasing method (ShelXT)46, refined with the ShelXL\u00a0program47, and expanded using Fourier techniques. All non-hydrogen atoms were refined anisotropically. Hydrogen atoms were included in structure factor calculations. All hydrogen atoms were assigned to idealized geometric positions.The crystal data of BCrystal data for B2IDip2(SBu)2(1a): The displacement parameters of atoms C63\u2013C74 were restrained to the same value with similarity restraint SIMU. C68H104B2N4S2, Mr\u2009=\u20091063.29, green block, 0.294\u2009\u00d7\u20090.257\u2009\u00d7\u20090.125\u2009mm3, monoclinic space group P21/c, a\u2009=\u200914.423(5)\u2009\u00c5, b\u2009=\u200912.522(5)\u2009\u00c5, c\u2009=\u200936.640(17)\u2009\u00c5, \u03b2\u2009=\u200996.25(3)\u00b0, V\u2009=\u20096578(5)\u2009\u00c53, Z\u2009=\u20094, \u03c1calcd\u2009=\u20091.074\u2009g\u2009cm\u20133, \u03bc\u2009=\u20090.122\u2009mm\u20131, F(000)\u2009=\u20092328, T\u2009=\u2009103(2)\u2009K, R1\u2009=\u20090.0980, wR2\u2009=\u20090.1550, 13420 independent reflections [2\u03b8\u2009\u2264\u200952.744\u00b0], and 768 parameters. CCDC-1567701.Crystal data for B2IDip2(SPh)2(1b): C66H82B2N4S2, Mr\u2009=\u20091017.09, red block, 0.25\u2009\u00d7\u20090.14\u2009\u00d7\u20090.05\u2009mm3, monoclinic space group P21/c, a\u2009=\u200922.2980(16)\u2009\u00c5, b\u2009=\u200924.4556(17)\u2009\u00c5, c\u2009=\u200942.972(3)\u2009\u00c5, \u03b2\u2009=\u200991.133(2)\u00b0, V\u2009=\u200923428(3)\u2009\u00c53, Z\u2009=\u200916, \u03c1calcd\u2009=\u20091.153\u2009g\u2009cm\u22123, \u03bc\u2009=\u20090.134\u2009mm\u20131, F(000)\u2009=\u20098768, T\u2009=\u2009100(2)\u2009K, R1\u2009=\u20090.1465, wR2\u2009=\u20090.1925, 48137 independent reflections [2\u03b8\u2009\u2264\u200952.746\u00b0], and 2730 parameters. CCDC-1567705.Crystal data for B2IDip2(SePh)2(1c): C66H82B2N4Se2, Mr\u2009=\u20091110.89, red block, 0.35\u2009\u00d7\u20090.15\u2009\u00d7\u20090.08\u2009mm3, monoclinic space group P21/c, a\u2009=\u200922.4904(11)\u2009\u00c5, b\u2009=\u200924.4964(13)\u2009\u00c5, c\u2009=\u200943.042(2)\u2009\u00c5, \u03b2\u2009=\u200991.335(2)\u00b0, V\u2009=\u200923707(2)\u2009\u00c53, Z\u2009=\u200916, \u03c1calcd\u2009=\u20091.245\u2009g\u2009cm\u20133, \u03bc\u2009=\u20091.291\u2009mm\u20131, F(000)\u2009=\u20099344, T\u2009=\u2009100(2)\u2009K, R1\u2009=\u20090.0804, wR2\u2009=\u20090.0917, 48482 independent reflections [2\u03b8\u2009\u2264\u200952.744\u00b0], and 2729 parameters. CCDC-1567706.Crystal data for B2CAAC2(SBu)2(2a): C51H87B2N2S2, Mr\u2009=\u2009813.96, orange block, 0.335\u2009\u00d7\u20090.257\u2009\u00d7\u20090.112\u2009mm3, triclinic space group Pa\u2009=\u200910.383(5)\u2009\u00c5, b\u2009=\u200914.048(9)\u2009\u00c5, c\u2009=\u200917.857(8)\u2009\u00c5, \u03b1\u2009=\u200975.299(15)\u00b0, \u03b2\u2009=\u200984.42(3)\u00b0, \u03b3\u2009=\u200977.545(19)\u00b0, V\u2009=\u20092458(2)\u2009\u00c53, Z\u2009=\u20092, \u03c1calcd\u2009=\u20091.100\u2009g\u2009cm\u20133, \u03bc\u2009=\u20090.143\u2009mm\u20131, F(000)\u2009=\u2009898, T\u2009=\u2009100(2)\u2009K, R1\u2009=\u20090.0532, wR2\u2009=\u20090.1087, 10053 independent reflections [2\u03b8\u2009\u2264\u200952.744\u00b0], and 533 parameters. CCDC-1567703.2CAAC2(SPh)2 (2b) and B2CAAC2(SePh)2 (2c) were collected on a Bruker D8 Quest diffractometer with a CMOS area detector and multi-layer mirror-monochromated MoK\u03b1 radiation. The structure was solved using the intrinsic phasing method (ShelXT)46, refined with the ShelXLprogram47, and expanded using Fourier techniques. All non-hydrogen atoms were refined anisotropically. Hydrogen atoms were included in structure factor calculations. All hydrogen atoms were assigned to idealized geometric positions.The crystal data of BCrystal data for B2CAAC2(SPh)2(2b): C52H72B2N2S2, Mr\u2009=\u2009810.85, orange block, 0.173\u2009\u00d7\u20090.161\u2009\u00d7\u20090.122\u2009mm3, monoclinic space group P21/n, a\u2009=\u20099.9798(15)\u2009\u00c5, b\u2009=\u200918.647(4)\u2009\u00c5, c\u2009=\u200924.919(5)\u2009\u00c5, \u03b2\u2009=\u200997.909(15)\u00b0, V\u2009=\u20094593.2(15)\u2009\u00c53, Z\u2009=\u20094, \u03c1calcd\u2009=\u20091.173\u2009g\u2009cm\u20133, \u03bc\u2009=\u20090.153\u2009mm\u20131, F(000)\u2009=\u20091760, T\u2009=\u2009100(2)\u2009K, R1\u2009=\u20090.0422, wR2\u2009=\u20090.0961, 9368 independent reflections [2\u03b8\u2009\u2264\u200952.746\u00b0], and 539 parameters. CCDC-1567702.Crystal data for B2CAAC2(SePh)2(2c): C52H72B2N2Se2, Mr\u2009=\u2009904.65, orange block, 0.061\u2009\u00d7\u20090.055\u2009\u00d7\u20090.025\u2009mm3, monoclinic space group P21/n, a\u2009=\u200910.060(7)\u2009\u00c5, b\u2009=\u200918.707(7)\u2009\u00c5, c\u2009=\u200924.857(14)\u2009\u00c5, \u03b2\u2009=\u200998.86(3)\u00b0, V\u2009=\u20094622(5)\u2009\u00c53, Z\u2009=\u20094, \u03c1calcd\u2009=\u20091.300\u2009g\u2009cm\u20133, \u03bc\u2009=\u20091.637\u2009mm\u20131, F(000)\u2009=\u20091904, T\u2009=\u2009100(2)\u2009K, R1\u2009=\u20090.0628, wR2\u2009=\u20090.0784, 9321 independent reflections [2\u03b8\u2009\u2264\u200952.746\u00b0], and 539 parameters. CCDC-1567704.48, B3LYP51, and BP8652 levels in conjunction with the def2-SVP53 basis set. The structures were characterized as minima by frequency calculation. The NBO54 analyses were carried out at the M05-2X/def2-SVP level calculations were carried out on diamagnetic diborenes 1a\u2013c and the diradicals 2a\u2013c in their singlet and triplet states whereas the triplet states of 2a-c are always more stable than the singlet states 43 approach to optimize the geometry of the lowest-lying singlet and triplet states of 2a. Both optimizations were started from the X-ray structure of the triplet ground state. The computed singlet geometry does not represent the global minimum of the singlet potential energy surface, which possesses a nearly planar CCAAC-B-B-CCAAC orientation. However, only the local minimum lying near the triplet equilibrium structure will not be populated in the measurements because the global minimum of the singlet state is separated from the triplet equilibrium geometry by high-energy barriers. For the computed geometries , the MN12L/6\u2013311G approach predicts an S\u2013T gap of about \u20136.4\u2009kcal\u2009mol\u20131 (triplet below singlet state).To obtain an estimate of the inaccuracies of DFT approaches, we first used the MN12L2a\u02b9 in which we replaced the bulky substituents as indicated in Supplementary Figure\u00a02a systems to include the influence of steric effects on the geometry. The model system 2a\u02b9 contains all essential binding ingredients of 2a but is sufficiently small to perform high-level NEVPT240/def2-TZVP benchmark calculations for the S\u2013T gap. To compare the predicted S\u2013T gaps, we performed single-point calculations employing the geometries described above.To enable high-level multi-reference approaches, we then created a closely related model system \u22121 to nearly \u201329\u2009kcal\u2009mol\u22121, showing that such DFT calculations have to be handled with care.The results are summarized in Supplementary Table\u00a0Cartesian coordinates for all of the calculated compounds are given in Supplementary Table\u00a02a were carried out with a Quantum-Design MPMS-XL-5 SQUID magnetometer equipped with a 5-Tesla magnet in the range from 295 to 2.0\u2009K at a magnetic field of 0.5\u2009T. The powdered sample was contained in a Teflon bucket and fixed in a non-magnetic sample holder. Each raw data file for the measured magnetic moment was corrected for the diamagnetic contribution of the Teflon bucket according to Mdia(bucket)\u2009=\u2009\u03c7g\u2009\u00d7\u2009m\u2009\u00d7\u2009H, with an experimentally obtained gram susceptibility of the Teflon bucket. The molar susceptibility data were corrected for the diamagnetic contribution using Pascal\u2019s constants56. Experimental data were modeled with the julX program57 using a fitting procedure to the spin Hamiltonian \u20136\u2009cm3\u2009mol\u20131) and some weak intermolecular interactions (\u20130.12\u2009K). The latter were considered in a mean field approach by using a Weiss temperature \u039858. The Weiss temperature \u0398 (defined as \u0398\u2009=\u2009zJinterS(S\u2009+\u20091)/3k) relates to intermolecular interactions zJinter, where Jinter is the interaction parameter between two nearest neighbor magnetic centers, k is the Boltzmann constant (0.695\u2009cm\u20131\u2009K\u20131), and z is the number of nearest neighbors.Temperature-dependent magnetic susceptibility measurements for http://www.ccdc.cam.ac.uk/data_request/cif.The data that support the findings of this study are available from the corresponding author on reasonable request. The X-ray crystallographic coordinates for structures reported in this study have been deposited at the Cambridge Crystallographic Data Centre (CCDC), under deposition numbers 1567701\u20131567706. These data can be obtained free of charge from The Cambridge Crystallographic Data Centre via Supplementary Information(PDF 4825 kb)Peer Review File(PDF 139 kb)Description of Additional Supplementary Files(PDF 166 kb)Supplementary Data 1(XLSX 196 kb)"} +{"text": "Scientific Reports7: Article number: 44813; 10.1038/srep44813 published online: 03202017; updated: 09202017.The original version of this Article contained typographical errors in the Abstract.6 (dielectric loss ~5\u2009\u00d7\u200910\u22127) at high powers, degrading to 7\u2009\u00d7\u200910\u22125 and g-factor of 1.995\u2009\u00b1\u20090.008\u201d.\u201cWhispering Gallery Mode (WGM) analysis revealed large Quality Factors of order 2\u2009\u00d7\u200910now reads:6 (dielectric loss ~5\u2009\u00d7\u200910\u22127) at high powers, degrading to 7\u2009\u00d7\u2009105 (dielectric loss ~1.4\u2009\u00d7\u200910\u22126) at single photon energy. A very low-loss narrow line width paramagnetic spin flip transition was detected with extreme sensitivity in 28Si, with very small concentration below 1011\u2009cm\u22123 (less than 10 parts per trillion) and g-factor of 1.995\u2009\u00b1\u20090.008\u201d.\u201cWhispering Gallery Mode (WGM) analysis revealed large Quality Factors of order 2\u2009\u00d7\u200910This has now been corrected in the PDF and HTML versions of the Article."} +{"text": "II with the bridging ligand bis\u00ad(pyridin-3-ylmeth\u00adyl)sulfane afforded a one-dimensional zigzag chain polymeric structure, with the charge balanced by two coordinated chloride anions. C\u2014H\u22efCl hydrogen bonds and Hg\u2014Cl\u22ef\u03c0 inter\u00adactions, together with C\u2014H\u22ef\u03c0 hydrogen bonds, stabilize the crystal structure.The reaction of Hg L, C12H12N2S) in methanol afforded the title crystalline coordination polymer catena-poly[[di\u00adchlorido\u00admercury(II)]-\u03bc-bis\u00ad(pyridin-3-ylmeth\u00adyl)sulfane-\u03ba2N:N\u2032], [HgCl2L]n. The asymmetric unit consists of one HgII cation, one L ligand and two chloride anions. Each HgII ion is coordinated by two pyridine N atoms from separate L ligands and two chloride anions. The metal adopts a highly distorted tetra\u00adhedral geometry, with bond angles about the central atom in the range 97.69\u2005(12)\u2013153.86\u2005(7)\u00b0. Each L ligand bridges two HgII ions, forming an infinite \u2013(Hg\u2013L)n\u2013 zigzag chain along the b axis, with an Hg\u22efHg separation of 10.3997\u2005(8)\u2005\u00c5. In the crystal, adjacent chains are connected by inter\u00admolecular C\u2014H\u22efCl hydrogen bonds, together with Hg\u2014Cl\u22ef\u03c0 inter\u00adactions [chloride-to-centroid distance = 3.902\u2005(3)\u2005\u00c5], that form between a chloride anion and the one of the pyridine rings of L, generating a two-dimensional layer extending parallel to (101). These layers are further linked by inter\u00admolecular C\u2014H\u22ef\u03c0 hydrogen bonds, forming a three-dimensional supra\u00admolecular network.The reaction of mercury(II) chloride with bis\u00ad(pyridin-3-ylmeth\u00adyl)sulfane ( The flexibility, length and coordinating ability of the spacer ligands exert strong influences on the formation of coordination polymers and their resulting diverse topologies (Zheng L), and has reported its AgI and CoII coordination polymers chloride with L sulfane sulfane, C12H12N2S. The asymmetric unit comprises one HgII cation, one L ligand and two chloride anions. The HgII ion is four-coordinated, binding to two Cl anions and two pyridine N atoms from two separate symmetry-related L ligands, forming a highly distorted tetra\u00adhedral geometry \u2005\u00c5. In the L ligand, the dihedral angle between the two terminal pyridine rings is 78.52\u2005(18)\u00b0, and the flexible thio\u00adether moiety [C4\u2013C6\u2013S1\u2013C7\u2013C8] shows a bent arrangement with a gauche--anti configuration . The conformation of the L ligand, along with its Npy\u2014Hg\u2014Npy coordination angle [98.39\u2005(16)\u00b0], may induce the zigzag topology of the chain.Fig.\u00a01ry Fig.\u00a01, with thry Fig.\u00a01. The S aon Fig.\u00a02. The sepet al., 2009L with Cl2\u22efCg1iv = 3.902\u2005(3)\u2005\u00c5 and Hg1\u2014Cl2\u22efCg1iv = 77.21\u2005(6)\u00b0 , generating layers extending parallel to (101). Neighboring layers are linked by C2\u2014H2\u22efCg2 hydrogen bonds gave three hits. Two of the L ligand adopts a bent arrangement that is similar to that of the HgII polymer described here. However, the title compound displays a zigzag topology and is the first example of an HgII coordination polymer with the ligand L.A search of the Cambridge Structural Database = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901701619X/sj5541sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S205698901701619X/sj5541Isup2.hklStructure factors: contains datablock(s) I. DOI: 1584773CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E)-4-meth\u00adoxy\u00adbenzyl\u00adidene\u00adamino]\u00adpropan-2-ol, mol\u00adecules are linked by O\u2014H\u22efN hydrogen bonds into C(5) supra\u00admolecular chains propagating along the a-axis direction.In the crystal of 1,3-bis\u00ad[\u00b0. In the crystal, O\u2014H\u22efN hydrogen bonds link the mol\u00adecules into infinite C(5) chains propagating along the a-axis direction. The packing of these chains is consolidated by C\u2014H\u22efO inter\u00adactions and C\u2014H\u22ef\u03c0 short contacts, forming a three-dimensional network.The title Schiff base, C The N1=C1 and N2=C5 distances of 1.265\u2005(4) and 1.271\u2005(4)\u2005\u00c5, respectively, are consistent with C=N double bonding. The bond angles of 117.0\u2005(3) and 117.7\u2005(3)\u00b0 around the N1 and N2 atoms confirm their sp2 character. The slight differences between N=C distances and C\u2014N=C angles are due to the significant effect of the hydrogen bond on the geometric parameters of the nitro\u00adgen atom (N2) involved in the inter\u00admolecular hydrogen bond (see below).The mol\u00adecular structure of the title compound is illustrated in Fig.\u00a01The N1\u2014C2\u2014C3\u2014C4 torsion angle is \u221264.4\u2005(4)\u00b0 and the C2\u2014C3\u2014C4\u2014N2 torsion angle is 175.0\u2005(3)\u00b0. The O1\u2014C3\u2014C4\u2014N2 torsion angle is \u221265.3\u2005(3)\u00b0, which has a significant role to play in the hydrogen-bonding pattern in the crystal of the title compound (see below). The two meth\u00adoxy substituents are essentially coplanar with their bound benzene rings with torsion angles C17\u2014O2\u2014C14\u2014C13 = 169.3\u2005(3)\u00b0 and C27\u2014O3\u2014C24\u2014C23 = \u2212172.2\u2005(3)\u00b0.C(5) chains extending along the a-axis direction. The chains are further linked to neighbouring chains through a pair of weak C\u2014H\u22efO hydrogen bonds (Table\u00a01Cg (\u03c0\u2013ring) inter\u00adactions = 0.95\u2005\u00c5 for aromatic and azomethine atoms, d(C\u2014H) = 0.99\u2005\u00c5 for methyl\u00adene, d(C\u2014H) = 1.00\u2005\u00c5 for C3\u2014H3 and 0.98\u2005\u00c5 for CH3 atoms. The Uiso values were constrained to be 1.5Ueq of the carrier atom for methyl H atoms and 1.2Ueq for the remaining H atoms. A rotating group model was used for the methyl groups. The absolute structure of the crystal chosen for data collection was indeterminate in the present refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016016947/hb7623sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016016947/hb7623Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016016947/hb7623Isup3.cmlSupporting information file. DOI: 1511139CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The synthesis and structural determination of a nickel(II) complex where the two nickel cations in the asymmetric unit each are coordinated by two tridentate, potentially redox non-innocent bis-imino\u00adpyridyl ligands are reported. N}eth\u00adyl)pyridine-\u03baN]nickel(II) dichloride\u2013di\u00adchloro\u00admethane\u2013water (1/2/2), [Ni(C25H29N5)2]Cl2\u00b72CH2Cl2\u00b72H2O, represents a nickel(II) bis\u00ad(pyridine di\u00adimine) complex with electron-donating di\u00admethyl\u00adamino\u00adphenyl substituents. The complex crystallizes as a water/di\u00adchloro\u00admethane solvate with Z\u2032 = 2, thus the asymmetric unit consists of two NiII complex cations, four chloride anions, four adventitious water and four di\u00adchloro\u00admethane solvent mol\u00adecules. Around each octa\u00adhedrally coordinated NiII cation, one pendant phenyl group on each of the two ligands has an intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adaction with the pyridine ring of the other chelating ligand. In the crystal, pairs of water mol\u00adecules are hydrogen bonded to pairs of chlorine atoms. The di\u00adchloro\u00admethane solvent mol\u00adecules are likewise hydrogen bonded to the chloride anions.The solvated title compound, bis\u00adimino-\u03ba A solution of 2,6-di\u00adacetyl\u00adpyridine , 4-(di\u00admethyl\u00adamino)\u00adaniline and formic acid (1\u2005ml) was prepared in toluene (100\u2005ml) under nitro\u00adgen atmosphere and then stirred for 12\u2005h on mol\u00adecular sieves. The reaction mixture was filtered and extracted with excess di\u00adchloro\u00admethane, then the amount of solvent was reduced in vacuo. The crude yellow product was then washed with cold methanol, followed by diethyl ether and filtered producing a pure bright-yellow solid . 1H NMR : \u03b4 8.4\u20138.2 , 7.9\u20137.8 , 6.8 3.0 , 2.5 . MS (ESI): 400.4 m/z [C25H29N5]H+.Synthesis of [bis-\u00adeth\u00adyl)pyridine)\u00adnickel(II)] chloride. A solution of the PDI-DMA ligand and nickel(II) chloride was prepared in THF (15\u2005ml) under nitro\u00adgen atmosphere, then stirred for 12\u2005h. The solution was filtered and extracted with di\u00adchloro\u00admethane. The solvent was removed in vacuo yielding a dark reddish-brown solid . X-ray diffraction quality crystals were isolated as red\u2013brown blocks by vapor diffusion of hexa\u00adnes into a saturated solution of the product and di\u00adchloro\u00admethane. The complex was NMR silent (paramagnetic). MS (ESI): m/z 855.5 [C50H57N10Ni]+.Uiso(H) = 1.2 \u00d7, 1.2 \u00d7 or 1.5 \u00d7 Ueq(C) for aromatic, methyl\u00adene and methyl H atoms, successively. Water H atoms were initially located from a difference Fourier map and included in their initially observed positions and allowed to ride with the position of the parent oxygen atom. Displacement parameters of the hydrogen atom were freely refined. Two reflections I. DOI: 10.1107/S2056989017010088/wm5402Isup2.hklStructure factors: contains datablock(s) I. DOI: 1560729CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A combined single-crystal X-ray diffraction and NMR crystallography study of a 1:1 cocrystal of two fungicides, namely di\u00adthia\u00adnon and pyrimethanil, is presented. Specifically, the role of hydrogen bonding and C\u2014H\u22ef\u03c0 and S\u22efO inter\u00admolecular inter\u00adactions is qu\u00adanti\u00adtatively investigated. H,10H-naphtho\u00addithiine-2,3-dicarbo\u00adnitrile\u20134,6-dimethyl-N-phenyl\u00adpyrimidin-2-amine (1/1), C14H4N2O2S2\u00b7C12H13N2]. Following an NMR crystallography approach, experimental solid-state magic angle spinning (MAS) NMR spectra are presented together with GIPAW (gauge-including projector augmented wave) calculations of NMR chemical shieldings. Specifically, experimental 1H and 13C chemical shifts are determined from two-dimensional 1H\u201313C MAS NMR correlation spectra recorded with short and longer contact times so as to probe one-bond C\u2014H connectivities and longer-range C\u22efH proximities, whereas H\u22efH proximities are identified in a 1H double-quantum (DQ) MAS NMR spectrum. The performing of separate GIPAW calculations for the full periodic crystal structure and for isolated mol\u00adecules allows the determination of the change in chemical shift upon going from an isolated mol\u00adecule to the full crystal structure. For the 1H NMR chemical shifts, changes of 3.6 and 2.0\u2005ppm correspond to inter\u00admolecular N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonding, while changes of \u22122.7 and \u22121.5\u2005ppm are due to ring current effects associated with C\u2014H\u22ef\u03c0 inter\u00adactions. Even though there is a close inter\u00admolecular S\u22efO distance of 3.10\u2005\u00c5, it is of note that the mol\u00adecule-to-crystal chemical shifts for the involved sulfur or oxygen nuclei are small.A single-crystal X-ray diffraction structure of a 1:1 cocrystal of two fungicides, namely di\u00adthia\u00adnon (DI) and pyrimethanil (PM), is reported [systematic name: 5,10-dioxo-5 While this is an established procedure in the development of new active pharmaceutical ingredients, where it is used to increase the solubility and bioavailability NMR with calculation of NMR parameters, is finding important application to moderately sized organic mol\u00adecules were mixed thoroughly in a 1:1 molar ratio (0.5\u2005g of pyrimethanil) and kept at 323\u2005K under agitation. After a couple of hours, the powdery product had changed to a dark-olive-green colour.The DI\u2013PM cocrystal was prepared according to method VII in point [0041] of Sowa al. 2013, i.e. drUiso(H) = 1.2\u20131.5Ueq(parent)], after which the positions were refined with riding constraints MAS experiments were performed on a Bruker Avance III spectrometer operating at 1H and 13C Larmor frequencies of 600 and 150.9\u2005MHz, respectively, using a 1.3\u2005mm HXY (1H MAS) or a 4\u2005mm HX (13C CP MAS) Bruker probe. In all cases, a 1H 90\u00b0 pulse duration of 2.5\u2005\u00b5s was used. 2D 1H\u201313C HETCOR experiments were performed on a Bruker Avance III spectrometer, using a 4\u2005mm HXY probe in double-resonance mode. In the HETCOR pulse sequence, the following phase cycling was employed: 1H 90\u00b0 pulse (90\u00b0 270\u00b0), 13C 180\u00b0 pulse (2{0\u00b0} 2{180\u00b0}), 13C CP contact pulse (4{0\u00b0} 4{180\u00b0} 4{90\u00b0} 4{270\u00b0}), receiver (0\u00b0 180\u00b0 0\u00b0 180\u00b0 180\u00b0 0\u00b0 180\u00b0 0\u00b0 90\u00b0 270\u00b0 90\u00b0 270\u00b0 270\u00b0 90\u00b0 270\u00b0 90\u00b0). For CP, a 70 to 100% ramp were relaxed and periodic boundary conditions were applied. The space group P21/n was preserved. All distances and angles stated in the main text of this article are for the geometry-optimized crystal structure. Note also that the geometry optimization within CASTEP causes a relabelling of the atoms \u2013 in this article, we use the CASTEP numbering; see Fig. S1 in the Supporting information for a comparison with the numbering employed in the crystallographic CIF file. The GIPAW method , a chain of mol\u00adecules is held together by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds (between DI and PM mol\u00adecules) and by putative S\u22efO inter\u00adactions and 1(c), respectively. As can be seen from the representation along the crystallographic a axis in Fig.\u00a01c), the packing is based on assemblies of blocks of four mol\u00adecules; four mol\u00adecules (PM\u2013DI\u2013DI\u2013PM) are arranged in a layer , forming a block that is perpendicular to an adjacent block of four mol\u00adecules, thus building up the \u2018zigzag\u2019 arrangement.The single-crystal X-ray diffraction structure of the DI\u2013PM cocrystal is schematically represented in Fig.\u00a01er Fig.\u00a01a, formi13C CP MAS NMR 1D spectrum of the DI\u2013PM cocrystal, together with three stick spectra that represent 13C chemical shifts calculated using the GIPAW method for the DI\u2013PM crystal structure. Specifically, the calculated 13C chemical shifts are presented in three groups according to whether they correspond to direct one-bond C\u2014H connectivities or nonprotonated C atoms . The distinction between Figs.\u00a02c) and 2(d) corresponds to whether cross peaks corresponding to a longer-range C\u22efH proximity are observed in 1H\u201313C 2D correlation spectra and 3(b) present 1H NMR spectra of the DI\u2013PM cocrystal recorded at a fast MAS frequency of 60\u2005kHz; specifically, a one-pulse one-dimensional spectrum in Fig.\u00a03a), together with vertical lines corresponding to calculated (GIPAW) 1H chemical shifts, as well as a 2D DQ spectrum in Fig.\u00a03b). In addition, Fig.\u00a03c) presents a 1H\u201313C 2D correlation spectrum of the DI\u2013PM cocrystal; note that this spectrum has been rotated through 90\u00b0 from its usual representation such that the direct (13C) dimension is vertical. In this way, it is possible to directly compare 1H chemical shifts of peaks in the 1H\u201313C and 1H DQ 2D and 1H 1D spectra. Two separate spectral regions are presented in Fig.\u00a03c) corresponding to (top) the methyl resonances at a 13C chemical shift close to 25\u2005ppm and (bottom) the aromatic CH resonances with 13C chemical shifts between 110 and 140\u2005ppm.Figs.\u00a033C Fig.\u00a03c and 1H2D Fig.\u00a03b and 1H1D Fig.\u00a03a spectr1H\u201313C correlation spectrum in Fig.\u00a03c) was recorded using a short CP contact time of 100\u2005\u00b5s to transfer magnetization from 1H to 13C, such that cross peaks correspond to one-bond C\u2014H connectivities. The spreading of the resonances into two dimensions in Fig.\u00a03c) allows the identification of two and ten resolved cross peaks for the CH3 and aromatic CH groups, respectively. The value of such a 1H\u201313C correlation spectrum in resolving and assigning the experimental 1H chemical shifts is thus evident. Table\u00a0213C chemical shifts . For directly bonded C\u2014H connectivities, H-atom labels and calculated (GIPAW) and experimental 1H chemical shifts are presented in normal font.The 1H\u201313C correlation spectra recorded with three different CP contact times of 100\u2005\u00b5s , 500\u2005\u00b5s and 1\u2005ms ; Fig.\u00a04a) is a copy of Fig.\u00a03c), but presented in the normal orientation, i.e. with the direct (13C) dimension horizontal. It is evident that additional cross peaks are observed for longer CP contact times \u2013 these correspond to longer-range C\u22efH proximities , 160.1 (atom C63) and 168.2\u2005ppm (atoms C64 and C67); these all correspond to intra\u00admolecular proximities within the di\u00adthia\u00adnon mol\u00adecule, i.e. C57 with H17 , H21 and H29 , C63 with H29 , C64 and C67 with H25 and CH3 protons . Of most inter\u00adest is the cross peak, which thus enables the determination of the NH 1H chemical shift.Fig.\u00a04\u00b5s Fig.\u00a04a, 500\u2005\u00b5\u00b5s Fig.\u00a04b and 1\u2005ms Fig.\u00a04c; Fig.\u00a01H chemical shifts assigned, let us re-examine the 1H DQ MAS spectrum in Fig.\u00a03b). In such a spectrum, cross peaks are observed in the DQ dimension at the sum of the two single-quantum (SQ) frequencies if there is a close proximity . Consider the two lowest-ppm aromatic CH protons H25 (4.0\u2005ppm) and H2 (6.2\u2005ppm) for which distinct 1H resonances are resolved in the 1H SQ dimension. For H25, the only DQ peak is at 4.0\u00a0+\u00a02.0 = 6.0\u2005ppm with the CH3 protons, since H25 is sandwiched between two methyl-group substituents on the PM mol\u00adecule. For H2, there is a DQ peak at 6.2\u00a0+\u00a07.5 = 13.7\u2005ppm corresponding to the intra\u00admolecular H\u22efH proximity with the neighbouring H1 and H3 DI aromatic CH protons, as well as a DQ peak at 6.2\u00a0+\u00a02.0 = 8.2\u2005ppm due to inter\u00admolecular proximities to the PM CH3 H atoms . Considering the high-ppm region, DQ cross peaks for the overlapping PI NH H29 (9.1\u2005ppm) and aromatic CH H17 (9.1\u2005ppm) resonances are observed at 9.1\u00a0+\u00a07.7 = 16.8\u2005ppm for intra\u00admolecular H29\u22efH21 (2.21\u2005\u00c5) and H17\u22efH18 (2.50\u2005\u00c5) proximities, as well as at 9.1\u00a0+\u00a02.0 = 11.1\u2005ppm for inter\u00admolecular proximities to PM methyl-group protons (closest distances of H17\u22efH26 = 2.48\u2005\u00c5 and H29\u22efH24 = 2.64\u2005\u00c5). For the other overlapping CH aromatic resonances, cross peaks due to intra\u00admolecular proximities with other CH aromatic resonances, as well as inter\u00admolecular proximities to the methyl protons, are also observed.With all the 1H\u201313C correlation spectra presented in Fig.\u00a0413C and 1H chemical shifts. Specifically, in Fig.\u00a04a), red crosses correspond to direct C\u2014H one-bond connectivities (C\u2014H distances under 1.2\u2005\u00c5), while in Figs.\u00a04b) and 4(c), red crosses are presented for C\u2014H proximities between 1.2 and 2.2\u2005\u00c5 , and between 2.2 and 3.0\u2005\u00c5 . We comment here on the level of agreement between experimental and calculated (GIPAW) chemical shifts. Starting with a consideration of the aromatic CH moieties ; this corresponds to the established observation that the discrepancy is within 1% of the chemical shift range . For the 1H chemical shifts, while most are within the usual 0.3\u2005ppm, some exhibit slightly larger discrepancies, notably 0.6\u2005ppm for atoms H17 and H25.In the \u2005\u00c5 Fig.\u00a04b, and b\u2005\u00c5 Fig.\u00a04c. We co3 groups , whereas the calculated 13C chemical shifts are both 8.5\u2005ppm lower than the experimental values, although the experimental difference in 13C chemical shifts between atoms C65 and C68 of 1.8\u2005ppm is reproduced by the calculation (difference of 1.9\u2005ppm). The explanation for this is well known, namely, the gradient of a plot of experimental 13C chemical shifts against calculated shielding deviates slightly from \u22121 , where the calculated 1H chemical shift of 10.5\u2005ppm is 1.4\u2005ppm higher than the experimental value of 9.1\u2005ppm. Such a large difference is explained by a known temperature dependence for hydrogen-bonded protons atom that is involved in an inter\u00admolecular N\u2014H\u22efO hydrogen bond to atom O1 . Inter\u00adestingly, \u0394\u03b4crystal\u2013mol\u00adecule = 2.0\u2005ppm for the aromatic CH H21 atom, for which Fig.\u00a01a) identifies an inter\u00admolecular C\u2014H\u22efO so-called weak hydrogen-bonding and H2 (\u22121.6\u2005ppm); as shown in Fig.\u00a051H chemical shift are a consequence of ring current effects associated with the proton pointing towards the centre of a six-membered aromatic ring of a nearby PM mol\u00adecule in a C\u2014H\u22ef\u03c0 inter\u00adaction, as has been noted previously in a number of other cases global, I. DOI: 10.1107/S2053229617000870/df3006Isup3.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2053229617000870/df3006sup2.txtCASTEP cif output. DOI: 10.1107/S2053229617000870/df3006sup4.txtmagres file. DOI: 10.1107/S2053229617000870/df3006sup5.pdfAdditional Tables 13C chemical shift values) and a Figure showing the difference in the numbering schemes between the crystallographic data and the output of the GIPAW (CASTEP) calculations. DOI: 1507863CCDC reference:"} +{"text": "Podanthus mitiqui, is reported and the stereochemical features established.The structure of erioflorin, a germacrane sesquiterpene lactone isolated from 19H24O6 cyclo\u00addeca\u00adfuran-9-yl methacrylate], is a tricyclic germacrane sesquiterpene lactone, which was isolated from Podanthus mitiqui (L.). The compound crystallizes in the space group P212121, and its mol\u00adecular structure consists of a methacrylic ester of a ten-membered ring sesquiterpenoid annelated with an epoxide and a butyrolactone. The structure is stabilized by one intramolecular C\u2014H\u22efO hydrogen bond. An O\u2014H\u22efO hydrogen bond and further C\u2014H\u22efO interactions can be observed in the packing.The title compound, erioflorin, C Podanthus mitiqui (Lindl) is an endemic plant of the Central Zone of Chile. It is an evergreen shrub that can reach up to two meters in height; its flowers are yellow or orange\u2013yellow globose inflorescences. Previous chemical investigations of extracts isolated from the stems and leaves of Podanthus mitiqui revealed the presence of sesquiterpene lactones with a germacrane framework such as ovatifolin, de\u00adacetyl\u00adovatifolin and arturin \u2005\u00c5 and smallest displacement parameters \u03c6 of 23.2\u2005(8), 252.2\u2005(3) and 346.1\u2005(2)\u00b0. The maximum deviation from the mean plane is 0.589\u2005(3)\u2005\u00c5 (C3). The C\u2014C bond lengths range from 1.474\u2005(5) to 1.557\u2005(4)\u2005\u00c5. The Z-configured double bond is located between C4 and C5 with a bond length of 1.326\u2005(4)\u2005\u00c5. Some bond angles differ notably from ideal values due to the ring strain, such as C3\u2014C4\u2014C5 and C4\u2014C5\u2014C6 . The bond angles within the ten-membered ring including Csp3 atoms range from 112.0\u2005(3)\u00b0 to 125.7\u2005(3)\u00b0. The ten-membered and the five-membered rings are trans-fused. The lactone ring shows a closed puckering on C6-C7 (twisted). The puckering amplitude and the smallest displacement parameter of the five-membered ring are q = 0.192\u2005(3)\u2005\u00c5 and \u03c6 = 58.7\u2005(9)\u00b0. With respect to the lactone ring, H6 and H7 are equatorially oriented, whereas the C6\u2014C5 and the C7\u2014C8 bonds are axial. The maximum deviations of the substituents from the best plane are 0.065\u2005(6)\u2005\u00c5 (O4) and \u22120.323\u2005(6)\u2005\u00c5 (C13). The 1,10-ep\u00adoxy ring is trans-fused. The C8 side chain is \u03b2 oriented as well as the C10 methyl group, whereas the C4 methyl group is \u03b1. The methacrylate substituent deviates from the planarity by twisting about C16\u2014C17 [torsion angle O5\u2014C16\u2014C17\u2014C19 = 28.4\u2005(5)\u00b0]. The structure is closely related to that of di\u00adhydro\u00adheliangine mono\u00adchlorido acetate , eriophyllin-B and eriophyllin-C , which were also isolated from Eriophyllum confertiflorum to 0.805\u2005(3)\u2005\u00c5, yielding a total puckering amplitude ii, running along the c-axis direction is formed via the hydroxyl group and the lactone oxo group , approximately between the a and b axes (C7\u2014H7\u22efO1i) and along b (C13\u2014H13B\u22efO6iii). Non-hydrogen inter\u00admolecular contacts are found between O2 and O4iv . The unit cell contains no residual solvent-accessible voids.The crystal structure features infinite chains connected by hydrogen bonds. A strong O\u2014H\u22efO hydrogen bond, namely O2\u2014H2\u22efO4up Fig.\u00a02. Furtheret al. . Aerial parts (9.6\u2005kg) were powdered and extracted by maceration with ethyl acetate for 3\u2005d. The organic layer was evaporated in vacuo giving a crude product (250\u2005g) which was further purified by column chromatography, giving a primary fractioning of 11 fractions (F1\u2013F11) by using increasing polarity from hexane to ethyl acetate. F-8 (6\u2005g) was further purified by column chromatography giving a white solid, which was recrystallized from EtOc, affording colourless crystals suitable for X-ray diffraction analysis. M.p. (from methanol): 499\u2013500\u2005K. For further physical data for erioflorin, see Torrance et al. , C\u2014H = 0.98\u2005\u00c5 (\u2013CH2), C\u2014H = 0.99\u2005\u00c5, (\u2013CH), C\u2014H = 0.94\u2005\u00c5 (=CH2), and Uiso(H) = 1.5Ueq(CH3) and Uiso(H) = 1.2Uiso, with the exception of the O\u2014H hydrogen atom, which was refined freely, but with Uiso(H) = 1.5Uiso(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017001700/zl2693sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017001700/zl2693Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017001700/zl2693Isup3.cmlSupporting information file. DOI: 1530526CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "As a result of the presence of these ordered species, the framework changes its symmetry from P4/mmm to P21/c. The 4,4\u2032-bi\u00adpyridine guest mol\u00adecules form chains inside the 6.5 \u00d7 6.9\u2005\u00c5 pores parallel to [100] in which the mol\u00adecules inter\u00adact through \u03c0\u2013\u03c0 stacking. Ordered water mol\u00adecules form infinite hydrogen-bonded chains inside a second pore system (1.6 \u00d7 5.3\u2005\u00c5 free aperture) perpendicular to the 4,4\u2032-bi\u00adpyridine channels.The coordination polymer {[Cu(SiF These inter\u00adconnected pores are filled with two other 4,4\u2032-bi\u00adpyridine mol\u00adecules and five water mol\u00adecules and this may be related to the fact that, in the present compound, guest mol\u00adecules fill the pores and inter\u00adact significantly with the framework atoms (see below).The 2D coordination grids are stacked along the [100] direction through the SiFs Figs. 3 and 4 \u25b8.62\u2212 show some variations : 45.29\u2005(7)\u00b0; bipy(N37\u2013N43): 30.31\u2005(7)\u00b0]. Whereas the 4,4\u2032-bi\u00adpyridine mol\u00adecules belonging to the coordination network are rather rigid between the metal atoms , the adsorbed 4,4\u2032-bi\u00adpyridine mol\u00adecules display significantly larger atomic displacement parameters [Ueq = 0.025\u2005(5)\u2005\u00c52].Four of the five water mol\u00adecules (O49 to O52) form infinite es Fig.\u00a05. The fifes Fig.\u00a05. The 4,4es Fig.\u00a05, and arees Fig.\u00a05; these iet al., 20166)(C10H8N2)2]n coordination polymer framework [CSD refcodes: GORWUF 2]\u00b78H2O}n undergoes a structural conversion when immersed in water, leading to an inter\u00adpenetrated network where SiF62\u2212 anions are shifted out of the coordination sphere of copper ions and are replaced by water mol\u00adecules [CSD refcodes: AFEHOI of hydrated copper(II) tetra\u00adfluorido\u00adborate was added to a refluxing aceto\u00adnitrile solution (5\u2005cm3) of 4,4\u2032-bi\u00adpyridine . After filtration, Et2O vapor was diffused into the mother liquor for seven days, and then the solvent was allowed to evaporate very slowly. A mixture of blue and violet crystals was obtained; whereas the diffraction spots of the blue crystals could not be properly indexed, the violet crystals were of very good quality and led to the structure reported on herein.An aqueous solution (5\u2005cmUiso(H) = 1.2Uiso(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989016016686/vn2118sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016016686/vn2118Isup2.hklStructure factors: contains datablock(s) I. DOI: 1510381CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(5) ring motif. In the crystals of both compounds, mol\u00adecules are linked by pairs of N\u2014H\u22efS hydrogen bonds, forming dimers with In the title compounds, the thio\u00adsemicarbazone group adopts an extended conformation, and there is a short N\u2014H\u22efN contact present forming an 11H13N3O2S, (I), and C16H15N3O2S, (II), the thio\u00adsemicarbazone group adopts an extended conformation. The acetate ester (I) crystallizes with two independent mol\u00adecules in the asymmetric unit. In the benzoate ester (II), the planes of the two aryl rings are inclined to one another by 46.70\u2005(7)\u00b0. In both compounds, there is a short intra\u00admolecular N\u2014H\u22efN contact present, forming an S(5) ring motif. In the crystals of both compounds, mol\u00adecules are linked via pairs of N\u2014H\u22efS hydrogen bonds, forming dimers with R22(8) ring motifs. The dimers are linked by N\u2014H\u22efS and N\u2014H\u22efO hydrogen bonds, forming slabs parallel to (01-1). In (I), there are N\u2014H\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions present within the slabs, while in (II), there are only N\u2014H\u22ef\u03c0 inter\u00adactions present.In the title compounds, C They display anti\u00adproliferative activity on different tumors cell lines and have been a common feature of all compounds with carcinogenic potency. A strong correlation has been found between tumor growth rate and the ribonucleoside diphos\u00adphate reductase (RDR) enzyme ring motif \u00b0 in mol\u00adecule B of compound (I)] and S1\u2014C16\u2014N2\u2014N1 [172.2\u2005(1)\u00b0 in compound (II)]. In compound (I)A and B, respectively]. The bond lengths C11A\u2014S1A [1.692\u2005(2)\u2005\u00c5] and C11B\u2014S1B [1.680\u2005(2)\u2005\u00c5] in (I)et al., 2016syn-periplanar to C5 [C5\u2014C6\u2014C7\u2014O1 = \u221215.8\u2005(2) \u00b0] and anti-periplanar to C1 [C1\u2014C6\u2014C7\u2014O1 = 160.7\u2005(1) \u00b0]. The dihedral angle between the benzene rings in compound (II)The mol\u00adecular structure of compounds (I)f Figs. 1 and 2 \u25b8.A\u2013B dimers with an In the crystal of (I)In the crystal of (II)f Table\u00a02. As in tet al., 2016E)-4-(N-carbamo\u00adthioyl\u00adethane\u00adhydrazono\u00adyl)phenyl 4-methyl\u00adbenzoate : Thio\u00adsemicarbazide was added to 50\u2005ml of an ethano\u00adlic solution of the 4-acetyl phenyl acetate (0.01\u2005mol) for (I)Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016018983/su5323sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989016018983/su5323Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016018983/su5323IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1519492, 1519491CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(6) ring motif. In the crystal, mol\u00adecules are linked by N\u2014H\u22efN and C\u2014H\u22efN hydrogen bonds forming chains propagating along the a-axis direction.The title isonicotinohydrazide derivative is planar, with an r.m.s. deviation for the fitted non-H atoms of 0.062\u2005\u00c5, and an intra\u00admolecular O\u2014H\u22efN hydrogen bond with an 13H10FN3O2, the mol\u00adecule has an E conformation with respect to the C=N bond of the hydrazone bridge. The dihedral angle between the isonicotinoyl and fluoro\u00adphenol moieties is 4.03\u2005(4)\u00b0, and an intra\u00admolecular O\u2014H\u22efN hydrogen bond generates an S(6) ring motif. In the crystal, mol\u00adecules are linked by N\u2014H\u22efN and C\u2014H\u22efN hydrogen bonds, forming chains propagating along the a-axis direction. The chains are linked by C\u2014H\u22efO hydrogen bonds, resulting in the formation of layers lying parallel to the ab plane. The crystal structure also features \u03c0\u2013\u03c0 inter\u00adactions [centroid-to-centroid distance = 3.6887\u2005(8)\u2005\u00c5].In the title compound, C The C6\u2014N2 and C7\u2014N3 bond lengths differ by 0.08\u2005\u00c5 hence; these two bonds are formally double and single bonds, respectively. The mol\u00adecule deviates slightly from planarity with an r.m.s deviation for the fitted non-hydrogen atoms of 0.062\u2005\u00c5. There is an intra\u00admolecular O2\u2014H2O\u22efN3 hydrogen bond with an S(6) ring motif present in the pyridine carboxamide moiety, and the pyridine ring (N1/C1\u2013C5) is approximately coplanar with the amide group (C6(=O1)N2) [dihedral angle = 8.25\u2005(6)\u00b0]. The isonicotinoyl moiety (N1/C1\u2013C6/O1/N2) is inclined to the fluoro\u00adphenol moiety (C8-C13/O2/F1) by 4.03\u2005(4)\u00b0.The mol\u00adecular structure of the title compound, with atom labelling, is presented in Fig.\u00a01N\u22efN1i and C4\u2014H4\u22efN1i hydrogen bonds \u2005\u00c5; Cg1 and Cg2 are the centroids of the pyridine (N1/C1\u2013C5) and phenyl (C8\u2013C13) rings, respectively; symmetry code: (i) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01].In the crystal, mol\u00adecules are linked by bifurcated-acceptor N2\u2014H2s Table\u00a01, leadinget al., 2016E)-N-(2-hy\u00addroxy\u00adbenzyl\u00adidene)isonicotinohydrazide skeleton revealed 86 hits. They include the isotypic crystal structures with bromide d 6.94 , 7.32 , 7.44 , 7.85 , 8.70 , 8.81 , 11.40 , 12.39 . HR\u2013MS (ES+) C13H11FN3O2 requires 260.0835 [M\u00a0+\u00a0H]+; found 260.0830.Isonicotinic acid hydrazide and 3-fluoro\u00adsalicyl\u00adaldehyde were suspended in a 1:1 mixture of water and ethanol (6\u2005ml). The reaction mixture was stirred at 363\u2005K for 24\u2005h and formation of a precipitate was observed. The reaction mixture was allowed to cool to room temperature and then filtered. The isolated solid was washed with water to give the product as a white solid . Colourless rod-like crystals, suitable for X-ray diffraction analysis, were grown by slow evaporation of a solution in methanol of the title compound. Uiso(H) = 1.5Ueq(O) and 1.2Ueq for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017009926/su5379sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017009926/su5379Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017009926/su5379Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989017009926/su5379Isup4.cmlSupporting information file. DOI: 1560196CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Cngb1 locus-encoded \u03b2-subunit of rod cGMP-gated cation channel and associated glutamic acid rich proteins (GARPs) are required for phototransduction, disk morphogenesis, and rod structural integrity. To probe individual protein structure/function of the GARPs, we have characterized several transgenic mouse lines selectively restoring GARPs on a Cngb1 knockout (X1\u2212/\u2212) mouse background. Optical coherence tomography (OCT), light and transmission electron microscopy (TEM), and electroretinography (ERG) were used to analyze 6 genotypes including WT at three and ten weeks postnatal. Comparison of aligned histology/OCT images demonstrated that GARP2 accelerates the rate of degeneration. ERG results are consistent with the structural analyses showing the greatest attenuation of function when GARP2 is present. Even 100-fold or more overexpression of GARP1 could not accelerate degeneration as rapidly as GARP2, and when co-expressed GARP1 attenuated the structural and functional deficits elicited by GARP2. These results indicate that the GARPs are not fully interchangeable and thus, likely have separate and distinct functions in the photoreceptor. We also present a uniform murine OCT layer naming nomenclature system that is consistent with human retina layer designations to standardize murine OCT, which will facilitate data evaluation across different laboratories.The The \u03b2-subunit and GARP2 have critical roles in phototransduction, disk morphogenesis, and maintenance of outer segment structural integrity1314Cngb1 locus, these proteins are identical in sequence for 318 amino acids. GARPs are intrinsically disordered proteins161615202215In the rod photoreceptor, the CNG channel is a heterotetramer composed of three \u03b1- and one \u03b2-subunits which are arranged around a central pore781011\u2212/\u2212), results in loss of function by one month, nearly undetectable levels of CNG \u03b1-subunit by two months, and retina degeneration by four months\u2212/\u2212) also results in loss of structure and function, including disruption of disk/plasma membrane interactions\u2212/\u2212 mice occurs 4\u20135 months earlier and the levels of the CNG \u03b1-subunit are significantly lower than in X1\u2212/\u2212 mice. To further dissect individual protein structure/function of the GARPs in rods, we have characterized several new transgenic mouse models, which selectively restore the absent GARP proteins to the X1\u2212/\u2212 mouse retina. Additionally, we present a system for murine optical coherence tomography (OCT) layer designation, that conforms with recently published human OCT guidelinesKnockout of the CNG channel \u03b2-subunit, while leaving GARPs intact and ten weeks (0.042\u2009mm\u2009\u00b1\u20090.01), but no change in FRT. Consistent with progressive retinal degeneration in X26\u2212/\u2212 mice, OCT analysis of transgenic mice showed significant reduction in ONL thickness (from 0.052\u2009mm\u2009\u00b1\u20090.01 to 0.033\u2009mm\u2009\u00b1\u20090.01) and FRT (from 0.215\u2009mm\u2009\u00b1\u20090.02 to 0.182\u2009mm\u2009\u00b1\u20090.01) over time. The effects of overexpression of G1 on the X1\u2212/\u2212 background were also assessed. X1\u2212/\u2212 G1Tg showed reduction in ONL thickness between three (0.052\u2009mm\u2009\u00b1\u20090.01) and ten weeks (0.038\u2009mm\u2009\u00b1\u20090.02), but no change in FRT. X1\u2212/\u2212 G2Tg showed a reduction in ONL thickness between three (0.051\u2009mm\u2009\u00b1\u20090.01) and ten weeks (0.019\u2009mm\u2009\u00b1\u20090.01), as well as a reduction in FRT (from 0.205\u2009mm\u2009\u00b1\u20090.02 to 0.163\u2009mm\u2009\u00b1\u20090.02). X1\u2212/\u2212 G1Tg G2Tg also showed a reduction in ONL thickness between three (0.050\u2009mm\u2009\u00b1\u20090.01) and ten weeks (0.028\u2009mm\u2009\u00b1\u20090.01), as well as a reduction in FRT (from 0.214\u2009mm\u2009\u00b1\u20090.01 to 0.176\u2009mm\u2009\u00b1\u20090.02). All of the genotypes except WT showed a reduction in ONL thickness over time. However, neither X1\u2212/\u2212 or X1\u2212/\u2212G1Tg showed a reduction in FRT, as was observed for all other genotypes.To assess the effects of the transgene alleles on retinal degeneration in X1en weeks and 4 in\u2212/\u2212 (0.212\u2009mm\u2009\u00b1\u20090.01), X1\u2212/\u2212 G1Tg (0.207\u2009mm\u2009\u00b1\u20090.02), X26\u2212/\u2212 (0.215\u2009mm\u2009\u00b1\u20090.02), X1\u2212/\u2212 G2Tg (0.205\u2009mm\u2009\u00b1\u20090.02), and X1\u2212/\u2212 G1Tg G2Tg (0.214\u2009mm\u2009\u00b1\u20090.01) all showed reduced FRT compared to WT (0.227\u2009mm\u2009\u00b1\u20090.02). At ten weeks X1\u2212/\u2212 and X26\u2212/\u2212 retinas showed reduced ONL and FRT thickness compared to WT . X26\u2212/\u2212 (0.033\u2009mm\u2009\u00b1\u20090.01) retinas showed reduced ONL thickness compared to X1\u2212/\u2212 (0.042\u2009mm\u2009\u00b1\u20090.01) and X1\u2212/\u2212 G1Tg (0.038\u2009mm\u2009\u00b1\u20090.02). X1\u2212/\u2212 G1Tg (0.038\u2009mm\u2009\u00b1\u20090.02) showed reduced ONL thickness compared to WT (0.051\u2009mm\u2009\u00b1\u20090.01) and X1\u2212/\u2212 (0.042\u2009mm\u2009\u00b1\u20090.01), as well as reduced FRT (0.205\u2009mm\u2009\u00b1\u20090.02) compared to WT (0.218\u2009mm\u2009\u00b1\u20090.02). X1\u2212/\u2212 G2Tg (0.019\u2009mm\u2009\u00b1\u20090.01) showed reduced ONL thickness compared to WT (0.051\u2009mm\u2009\u00b1\u20090.01), X1\u2212/\u2212 (0.042\u2009mm\u2009\u00b1\u20090.01), X26\u2212/\u2212 (0.033\u2009mm\u2009\u00b1\u20090.01), X1\u2212/\u2212 G1Tg (0.038\u2009mm\u2009\u00b1\u20090.02) and X1\u2212/\u2212 G1Tg G2Tg (0.028\u2009mm\u2009\u00b1\u20090.01), as well as reduced FRT (0.163\u2009mm\u2009\u00b1\u20090.02) compared to WT (0.218\u2009mm\u2009\u00b1\u20090.02), X26\u2212/\u2212 (0.182\u2009mm\u2009\u00b1\u20090.01) and X1\u2212/\u2212 G1Tg (0.205\u2009mm\u2009\u00b1\u20090.02). X1\u2212/\u2212 G1Tg G2Tg (0.028\u2009mm\u2009\u00b1\u20090.01) showed reduced ONL thickness compared to WT (0.051\u2009mm\u2009\u00b1\u20090.01), X1\u2212/\u2212 (0.042\u2009mm\u2009\u00b1\u20090.01), X26\u2212/\u2212 (0.033\u2009mm\u2009\u00b1\u20090.01), and X1\u2212/\u2212 G1Tg (0.038\u2009mm\u2009\u00b1\u20090.02), as well as reduced FRT (0.176\u2009mm\u2009\u00b1\u20090.02) compared to WT (0.218\u2009mm\u2009\u00b1\u20090.02) and X1\u2212/\u2212 G1Tg (0.205\u2009mm\u2009\u00b1\u20090.02). Genotypes expressing GARP2 in the absence of \u03b2-subunit had significant reduction in ONL thickness compared to those that did not express GARP2 (X1\u2212/\u2212 and X1\u2212/\u2212 G1Tg) or expressed GARP2 and \u03b2-subunit (WT).OCT also revealed between group differences in ONL thickness and FRT at each time point . At thre\u2212/\u2212, X26\u2212/\u2212 and X1\u2212/\u2212 G1Tg than in WT (p\u2009<\u20090.001). ONL thinned more rapidly in X1\u2212/\u2212 G2Tg than in WT, X1\u2212/\u2212, X26\u2212/\u2212, X1\u2212/\u2212 G1Tg, or X1\u2212/\u2212 G1Tg G2Tg. ONL thinned more rapidly in X1\u2212/\u2212 G1Tg G2Tg than in WT, X1\u2212/\u2212, or X1\u2212/\u2212 G1Tg. Therefore, the greatest rate of ONL thinning is in X1\u2212/\u2212 G2Tg. There is ONL thinning in X1\u2212/\u2212 G1Tg, but at a much slower rate than in X1\u2212/\u2212 G2Tg. Expression of both GARP1 and GARP2 in X1\u2212/\u2212 G1Tg G2Tg shows a slower rate of ONL thinning than X1\u2212/\u2212 G2Tg, but faster than X1\u2212/\u2212 G1Tg. Thus, in order of decreasing rates of thinning, the comparative rate of thinning per genotype is X1\u2212/\u2212 G2\u2009>\u2009X1\u2212/\u2212 G1Tg G2Tg\u2009>\u2009X26\u2212/\u2212\u2009>\u2009X1\u2212/\u2212 G1Tg\u2009>\u2009X1\u2212/\u2212\u2009>\u2009WT. The only difference between X26\u2212/\u2212 and X1\u2212/\u2212 G1Tg G2Tg, is that the latter expresses more GARP1 than the former. This indicates that co-expression of GARP2 along with increased GARP1 (X1\u2212/\u2212 G1Tg G2Tg) results in increased rates of thinning. However, increased expression of GARP1 without GARP2 (X1\u2212/\u2212 G1Tg) has a lower rate of thinning than endogenous expression of GARP1 with GARP2 (X26\u2212/\u2212), suggesting that GARP2 expression has the strongest effect on the rate of thinning.The rate of thinning over time was compared between groups, a measurement of the interaction between time and genotype . In gene\u2212/\u2212 and X26\u2212/\u2212 than in WT or X1\u2212/\u2212 G1Tg. The rate of FRT thinning is greater in X1\u2212/\u2212 G2Tg than WT or X1\u2212/\u2212 G1Tg. Finally, the rate of FRT thinning is greater in X1\u2212/\u2212 G1Tg G2Tg than WT or X1\u2212/\u2212 G1Tg. Considering all genotypes, the rate of thinning from fastest to slowest is X1\u2212/\u2212 G2Tg, X1\u2212/\u2212 G1Tg G2Tg, X1\u2212/\u2212, and X26\u2212/\u2212\u2009>\u2009X1\u2212/\u2212 G1Tg and WT. The only genotype that did not have faster FRT thinning than WT was X1\u2212/\u2212 G1Tg.The rate of FRT thinning is great\u2212/\u2212, 0.039\u2009mm\u2009\u00b1\u20090.005 for X26\u2212/\u2212, 0.040\u2009mm\u2009\u00b1\u20090.003 for X1\u2212/\u2212 G1Tg, 0.042\u2009mm\u2009\u00b1\u20090.003 for X1\u2212/\u2212 G2Tg and 0.037\u2009mm\u2009\u00b1\u20090.003 for X1\u2212/\u2212 G1Tg G2Tg. At ten weeks WT ONL is 0.036\u2009mm\u2009\u00b1\u20090.002, compared to 0.032\u2009mm\u2009\u00b1\u20090.003 for X1\u2212/\u2212, 0.026\u2009mm\u2009\u00b1\u20090.003 for X26\u2212/\u2212, 0.029\u2009\u00b1\u20090.004 for X1\u2212/\u2212 G1Tg, 0.021\u2009mm\u2009\u00b1\u20090.002 for X1\u2212/\u2212 G1Tg G2Tg, and 0.025\u2009mm\u2009\u00b1\u20090.004 for X1\u2212/\u2212 G2. Similarly to OCT results, there was a significant within group difference between 3 and 10 weeks for all genotypes except WT and X1\u2212/\u2212. While OCT results did show a significant thinning for X1\u2212/\u2212 between 3 and 10 weeks, this was the smallest difference and the least statistically significant change of all of the genotypes. Additionally, there were between group differences for ONL thickness at 10 weeks. OCT results showed a significant thinning in ONL at 10 weeks in X26\u2212/\u2212, X1\u2212/\u2212 G1Tg G2Tg, and X1\u2212/\u2212 G2Tg retinas compared to WT. However, histology measurements showed a significant thinning in ONL at 10 weeks for all genotypes compared to WT. At 3 weeks, WT FRT is 0.182\u2009mm\u2009\u00b1\u20090.006, compared to 0.179\u2009mm\u2009\u00b1\u20090.005 for X1\u2212/\u2212, 0.245\u2009mm\u2009\u00b1\u20090.025 for X26\u2212/\u2212, 0.233\u2009mm\u2009\u00b1\u20090.013 for X1\u2212/\u2212 G1Tg, 0.220\u2009mm\u2009\u00b1\u20090.012 for X1\u2212/\u2212 G2Tg, and 0.200\u2009mm\u2009\u00b1\u20090.012 for X1\u2212/\u2212 G1Tg G2Tg. At 10 weeks, WT FRT is 0.205\u2009mm\u2009\u00b1\u20090.010, compared to 0.197\u2009mm\u2009\u00b1\u20090.014 for X1\u2212/\u2212, 0.190\u2009mm\u2009\u00b1\u20090.013 for X26\u2212/\u2212, 0.179\u2009mm\u2009\u00b1\u20090.011 for X1\u2212/\u2212 G1Tg, 0.201\u2009mm\u2009\u00b1\u20090.020 for X1\u2212/\u2212 G2Tg, and 0.155\u2009mm\u2009\u00b1\u20090.006 for X1\u2212/\u2212 G1Tg G2Tg. While OCT results only showed a significant FRT thinning over time in WT, X26\u2212/\u2212, X1\u2212/\u2212 G2Tg, and X1\u2212/\u2212 G1Tg G2Tg, histology measurements showed significant FRT thinning over time in all genotypes. OCT results at 10 weeks showed a significantly thinner FRT for X26\u2212/\u2212, X1\u2212/\u2212 G1Tg, X1\u2212/\u2212 G1Tg G2Tg, and X1\u2212/\u2212 G2Tg compared to WT. Similar results were observed for histology measurements, except histology did not show a difference in FRT between WT and X26\u2212/\u2212 retinas. Overall, histology measurements of ONL and FRT confirmed OCT observations. In addition, histology also showed that photoreceptor outer segments are less uniform and more disorganized in X1\u2212/\u2212, X1\u2212/\u2212 G1Tg and X1\u2212/\u2212 G1Tg G2Tg. Photoreceptor outer segments are almost completely lost in X1\u2212/\u2212 G2Tg at 10 weeks. At 10 weeks, GARP2 accelerates thinning; however GARP1 does not affect the rate of thinning and partially counteracts the effect of GARP2 in the X1\u2212/\u2212 G1Tg G2Tg hybrid.To validate OCT results, histologic analysis was performed on each genotype at three and ten weeks . At thre\u2212/\u2212, X1\u2212/\u2212 G1Tg, X1\u2212/\u2212 G2Tg, and X1\u2212/\u2212 G1Tg G2Tg retinas images revealed structural changes as early as three weeks in X1 retinas . In WT, Cngb1) at 1 month postnatal , without the N-Cngb1 primary antibody showed no signal in any genotype.Because there is an ongoing degeneration it could not be assumed that the transgene expressed GARPs are properly localizing in photoreceptors. Therefore, we determined the localization of GARPs in WT and the transgenic animals. Using an antibody that recognizes the N-terminus of the \u03b2-subunit and GARPs . When GARP1 is overexpressed without GARP2 (X1\u2212/\u2212G1Tg) or co-expressed with GARP2 (X1\u2212/\u2212G1TgG2Tg) it was readily apparent. GARP2 was easily detected in WT, X1\u2212/\u2212G2Tg, X1\u2212/\u2212G1TgG2Tg, and X26\u2212/\u2212 mice. Protein expression is shown as the percent of GARP2 expression in WT retina in X1\u2212/\u2212G1Tg mice and 215% (\u00b126%) in X1\u2212/\u2212G1TgG2Tg mice. GARP2 was expressed at 42% (\u00b125%) in X1\u2212/\u2212G2Tg mice, 204% (\u00b172%) in X1\u2212/\u2212G1TgG2Tg mice, and 18% (\u00b112%) in X26\u2212/\u2212 mice. Despite a 30% increase in GARP1 expression in X1\u2212/\u2212G1TgG2Tg compared to X1\u2212/\u2212G1Tg mice, this difference was not statistically significant. However, there was significantly lower expression of GARP2 in X1\u2212/\u2212G2Tg and X26\u2212/\u2212 mice compared to X1\u2212/\u2212G1TgG2Tg mice. Despite a 20% increase in GARP2 expression in X1\u2212/\u2212G2Tg mice compared to X26\u2212/\u2212 mice, this difference was not statistically significant.To compare proteins levels of channel \u03b2-subunit, GARP1, and GARP2 in the various genotypes, automated capillary-based Western analysis was performed, using an internal standard for normalization. Similar to previously published resultsT retina , lane WTT retina . While G2 flash were significantly higher than X1\u2212/\u2212 , X26\u2212/\u2212 , X1\u2212/\u2212 G1Tg , X1\u2212/\u2212 G1Tg G2Tg and X1\u2212/\u2212 G2Tg . In addition, the X1\u2212/\u2212 a-wave was significantly higher than X1\u2212/\u2212 G2Tg, and the b-wave was significantly higher than X1\u2212/\u2212 G2Tg and X1\u2212/\u2212 G1Tg G2Tg. At 10 weeks , X26\u2212/\u2212 , X1\u2212/\u2212 G1Tg , X1\u2212/\u2212 G1Tg G2Tg and X1\u2212/\u2212 G2Tg . These data are consistent with a negative effect of GARP2 on rod function in the absence of the \u03b2-subunit. Scotopic responses were also measured at lower intensities (0.0001\u201310\u2009cd*s/m2) and showed similar results as the saturating 25\u2009cd*s/m2 flash .Using the mouse OCT nomenclature described in \u2212/\u2212 G2Tg mice. Co-expression of GARP1 and GARP2 on the X1\u2212/\u2212 reduced this attenuation, but it was still greater than the attenuation observed in X1\u2212/\u2212 G1Tg. These results provide further evidence that the negative effect of GARP2 is dominant over GARP1, and suggests a possible interaction between GARP1 and GARP2. This also further supports the conclusion that GARP2, in the absence of the \u03b2-subunit, drives functional and structural decline in the retina. Finally, coupled with the structural data, this also suggests GARP1 and GARP2 have distinct functions in the rods.As expected, photoreceptor (a-wave) and bipolar cell function (b-wave), measured via ERG, were reduced in all genotypes compared to WT. In addition, photoreceptor and bipolar cell function (attenuated a- and b-waves in ERG) were consistent with OCT findings, with the greatest attenuation in X1\u2212/\u2212 background which increased the GARP1 levels to approximately endogenous levels of GARP2 expression. A GARP2 transgene (at endogenous levels of protein when expressed on a WT background) was also crossed onto the X1\u2212/\u2212 background. In addition, the two GARP expressing animals were then crossed to enable examination of expression of the two transgenes in the absence of the CNG \u03b2-subunit. Finally, endogenous GARP1 and GARP2 were both expressed in the X26\u2212/\u2212 mouse which only lacks the CNG \u03b2-subunit. If the two proteins have identical function one would expect identical phenotypes when expressed in various genetic backgrounds where phenotypes are discernible. Furthermore, one would expect the co-expression of both GARPs to further accelerate the thinning. The ONL in X26\u2212/\u2212 mice thins more slowly than in X1\u2212/\u2212 G2Tg.In WT mice, GARP1 is 20-fold less abundant than GARP2 and four-fold less than the CNG \u03b2-subunit\u2212/\u2212, X26\u2212/\u2212, and a G2 overexpression mouse131419\u2212/\u2212, and the G2 overexpression mouse, GARP/\u03b2-subunit expression is confined to the OS with variable OPL signal. Unsurprisingly, GARP/\u03b2-subunit expression is absent in the X1\u2212/\u2212 mouse. GARP expression in the X1\u2212/\u2212 G1Tg, X1\u2212/\u2212 G2Tg, and X1\u2212/\u2212 G1Tg G2Tg mice was confined primarily to the OS, with minimal expression in the IS and variable signal in OPL. This shows that the transgene, even when overexpressed, primarily localizes to the OS.Previous studies have examined GARP/\u03b2-subunit expression in WT, X1\u2212/\u2212 mice is ~18% of the endogenous GARP2 expression, while GARP2 expression in X1\u2212/\u2212 G2Tg mice is ~42%. This higher level of GARP2 expression could explain why the ONL in X26\u2212/\u2212 mice thins more slowly than in X1\u2212/\u2212 G2Tg. Crossing the X1\u2212/\u2212 G1Tg (185% of WT GARP2 expression) and X1\u2212/\u2212 G2Tg (42% of WT GARP2 expression) mice results in offspring that express GARP2 at ~200% of WT GARP2 levels. X1\u2212/\u2212 G1Tg mice do not have significantly faster ONL thinning than X26\u2212/\u2212, but the X1\u2212/\u2212 G1Tg G2Tg mice (with the 200% increases in GARP2 expression) do have significantly faster rates of thinning than X26\u2212/\u2212 mice. However, the increased GARP2 expression in X1\u2212/\u2212 G1Tg G2Tg is not enough to increase the rate of thinning compared to X1\u2212/\u2212 G2Tg mice, which have lower levels of GARP2. Thus, the presence of GARP1 must be slowing the rate of thinning. This also indicates differing roles for GARP1 and GARP2 and the possibility that increased expression of GARP1 is somehow involved in increasing expression of GARP2. Taken together, our results lead us to conclude that GARP1 and GARP2 are not interchangeable as they do not produce identical phenotypes, nor are they additive in effect, and thus these proteins may have distinct and separate roles in the rod photoreceptor.Total GARP2 protein expression in X26\u2212/\u2212\u2212/\u2212One possible mechanism for the acceleration of degeneration in our GARP transgenic mice could be inhibition of \u03b1-subunit transport (in the absence of the \u03b2-subunit) from IS to OS which is consistent with the significantly lower \u03b1-subunit levels found in X26\u2212/\u2212)\u2212/\u2212)Tg, 100-fold overexpressed) using a 4.4\u2009kb opsin promoter, mouse protamine poly-A construct generated by replacing the PDE6 \u03b3-subunit insertTg, construct shown in \u2212/\u2212 G1Tg G2Tg mice carrying both alleles were generated by crossing G1Tg and G2Tg transgenic mice on a homozygous X1\u2212/\u2212 background and identifying the presence of both alleles by PCR genotyping (see below). All animal lines were made congenic on a C57BL/6J background. All animal use protocols were approved by the University of Alabama at Birmingham (UAB) Institutional Animal Care and Use Committee and are consistent with the Association for Research in Vision and Ophthalmology guidelines for the use of animals in research. All animals were maintained on a standard 12/12-hour light/dark cycle, fed standard rodent chow, and housed with standard rodent bedding.An exon 1, 2 and predicted promoter Cngb1 knockout null allele in digestion buffer consisting of 5\u2009mM EDTA, 200\u2009mM NaCl, 100\u2009mM Tris-HCl, and 0.2% SDS. Tails were digested overnight at 55\u2009\u00b0C. 100% ethanol was added to the lysate, which was then centrifuged at 16,000\u2009\u00d7\u2009g for thirty minutes, washed with 70% ethanol, centrifuged at 16,000\u2009\u00d7\u2009g for twenty minutes, and then the DNA pellet was dissolved in TE buffer, pH 8.0. PCR was performed using the following cycling parameters: 94\u2009\u00b0C denaturation, five minutes, for one cycle; thirty five cycles of 94\u2009\u00b0C, thirty seconds; for GARP1, 58\u2009\u00b0C, thirty seconds; for GARP2, 62\u2009\u00b0C, forty five seconds; 72\u2009\u00b0C, fifty seconds; and one cycle of 72\u2009\u00b0C for seven minutes. Primers sequences for X26TM 1.4 software rectangular volume scan of 1.4\u2009mm diameter with one thousand A-scans/B-scan, one hundred B-scans/volume, and one frame/B-scan or (scan parameter two) rectangular volume scan of 1.6\u2009mm in diameter with one thousand A-scans/B-scan, three B-scans/volume, and forty eight frames/B-scan. The one thousand A-scan frames collected in parameter two were averaged to reduce noise and increase signal. These averaged images were then converted to bmp files. To determine the ONL thickness, the difference in distance between the outer plexiform layer (OPL) and the external limiting membrane (ELM) was calculated. To determine the FRT, the difference in distance between the inner limiting membrane (ILM) and retinal pigment epithelium (RPE) was calculated. Measurements of ONL thickness and FRT were performed with Bioptigen Diver 2.4 software on retinal OCT images, captured using scan parameter one, of three week and ten week old mice at eight equidistant eccentricities was performed to visualize the posterior segment in living mice. The Bioptigen OCT instrument has a transverse resolution of 2.5\u2009\u03bcm and an axial resolution of 1.6\u2009\u03bcm. Animals were anesthetized with intraperitoneal injection of ketamine/xylazine at 100\u2009mg/kg and 10\u2009mg/kg body weight, respectively. Pupils were dilated using 1% tropicamide and 0.5% proparacaine , and artificial tears were applied frequently, with saline wash throughout to maintain corneal clarity. Scans were captured with the beam centered on the optic nerve. Both horizontal and vertical linear scans were obtained with Bioptigen InVivoVuericities , in one ricities . ONL thiricities were takLinear regression mixed models were used to assess ONL and FRT in five groups representing each transgenic line and WT mice. Linear regression mixed models were used to account for the within-animal correlation that occurs when multiple observations are taken from the same subject. First, we examined whether the mean ONL and FRT across the entire retina differed within each transgenic line group over time (three weeks vs. ten weeks). T-tests were also used to examine this association at each eccentricity . Next, we examined whether there were any between-group differences at each time point. Finally, a statistical interaction term was added to the model to assess if the changes over time differed by transgenic group. Separate linear mixed models were used to assess mean ONL and mean FRT. For five of the animals, data was collected from the same animal at both time points. Therefore, two separate analyses were employed to account for the paired nature of these measurements. First, data collected at ten weeks was treated as independent observations from those collected at time three. Second, the analyses were re-run excluding data collected at ten weeks for just those five animals. In both approaches, the pattern of results was similar. The results from the first analysis are presented; however, differences in results are noted where applicable. Adjustment of p-values for multiple comparisons was not performed. P-values\u2009<\u20090.05 were considered statistically significant.Cngb1; QEPPEPKDPPKPPGC) (1:50)Morphologic analysis was performed as previously describedCngb1; QEPPEPKDPPKPPGC) was used (1:500) to detect expression of the \u03b2-subunit and GARP2. N-Cngb1 was generated in rabbit and affinity purified . Data analysis was performed using the Compass Software (ProteinSimple) and quantitation was determined by normalizing the area under the curve of the \u03b2-subunit and GARP2 peaks by the area under the curve of the ProteinSimple internal standard peak. Representative computer generated electerophoretic images were generated by the Compass Software. Images shown contain information from capillaries run over multiple trials. The computer generated electrophoretic images displayed larger product size than is normally observed for these proteins. The very low pI of the proteins is likely the basis for the altered migration, which is even more pronounced in the capillary than is observed in an acrylamide gelWestern analysis on blots for GARP1 were performed as previously described2 and 25\u2009cd*s/m2 were delivered through a Ganzfeld dome on an Ocuscience ERG instrument manufactured for functional analysis of animal eyes. ERG data, including time/response and waveforms, were viewed using proprietary ERGView 4.860\u2009A software and analyzed after extraction into Microsoft Excel 2013 (ver. 15.0.4763.1003). Standard t-tests were used to analyze differences in the maximum saturating a- and b-wave response (\u03bcV) between genotypes at each light intensity.Scotopic ERG responses were measured at 6 different intensities of light in each genotype at 3 weeks and 10 weeks of age. Mice to be used for full-field ERG analysis were dark adapted overnight, sedated with 3% isoflurane in a closed chamber, and anesthesia was maintained at 2% via a certified isoflurane vaporizer . Eyes were anesthetized with proparacaine drops and pupils dilated with topical phenylephrine HCL and tropicamide . Only eyes receiving light stimuli were dilated. Methylcellulose 2.5% was applied to the corneal surface as well as to the contact lens placed on top of the silver-embedded recording electrode. The reference and ground electrodes were stainless steel subdermal needles placed in the cheek and lower back, respectively. During recordings the equipment and animal were enclosed in a Faraday cage and body temperature maintained at 37\u2009\u00b0C by a digital heating pad placed under the animal. Defined light pulses between 0.1\u2009cd*s/mHow to cite this article: DeRamus, M. L. et al. GARP2 accelerates retinal degeneration in rod cGMP-gated cation channel \u03b2-subunit knockout mice. Sci. Rep.7, 42545; doi: 10.1038/srep42545 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations."} +{"text": "The di\u00adhydro\u00adpyrimidine ring adopts a twist-boat conformation while the quinone ring is slightly non-planar. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions. In addition, a short inter\u00admolecular S\u22efN contact occurs. 24H18N2O4S, crystallizes in the ortho\u00adrhom\u00adbic P212121 space group, indicating the existence of only one enanti\u00adomer with an S configuration of the chiral center in the crystal phase. The di\u00adhydro\u00adpyrimidine ring adopts a twist-boat conformation while the quinone ring is slightly non-planar. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions. In addition, a short inter\u00admolecular S\u22efN contact of 3.250\u2005(3)\u2005\u00c5 indicates an inter\u00adaction between the S atom and the \u03c0-system of the thia\u00adzole ring.The title compound, C It was expected that the reaction would proceed with substitution of one or both chlorine atoms and the formation of a thia\u00adzole ring in the product 3 compounds that can be modified easily depending on the starting materials used during the reaction and 0.458\u2005(2)\u2005\u00c5, respectively. Such a conformation is common for 1,6-di\u00adhydro\u00adaromatic heterocycles and \u2212160.5\u2005(3)\u00b0, respectively].The di\u00adhydro\u00adpyrimidine ring adopts a twist-boat conformation with puckering parameters contact of 3.250\u2005(3)\u2005\u00c5 indicates an inter\u00adaction between the S atom and the \u03c0-system of the thia\u00adzole ring was added to a solution of 2,3-di\u00adchloro\u00adnaphthalene-1,4-dione in DMF (20\u2005mL) and the mixture was kept under reflux for 3\u2005h. After that, the reaction mixture was cooled, and the precipitated solid product was filtered off and purified via recrystallization from MeOH:DMF:H2O (2:2:1) to give product 3 in the form of dark-red crystals in 78% yield (0.35\u2005g), m.p. 520.3\u2013 522.0\u2005K.Ethyl 6-methyl-4-phenyl-2-thioxo-1,2,3,4-tetra\u00adhydro\u00adpyrim\u00adid\u00adine-5-carboxyl\u00adate Uiso(H) = nUeq(C) .Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018000117/zq2240sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018000117/zq2240Isup2.hklStructure factors: contains datablock(s) I. DOI: 1814348CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "EXE) is an endocrine therapy commonly used by postmenopausal women with hormone\u2010responsive breast cancer due to its potency in inhibiting aromatase\u2010catalyzed estrogen synthesis. Preliminary in\u00a0vitro studies sought to identify phase I EXE metabolites and hepatic cytochrome P450s (CYP450s) that participate in EXE biotransformation. Phase I metabolites were identified by incubating EXE with HEK293\u2010overexpressed CYP450s. CYP450s 1A2, 2C8, 2C9, 2C19, 2D6, 3A4, and 3A5 produce 17\u03b2\u2010dihydroexemestane (17\u03b2\u2010DHE), an active major metabolite, as well as two inactive metabolites. 17\u03b2\u2010DHE formation in pooled human liver microsomes subjected to isoform\u2010specific CYP450 inhibition was also monitored using tandem mass spectrometry. 17\u03b2\u2010DHE production in human liver microsomes was unaffected by isoform\u2010specific inhibition of CYP450s 2A6, 2B6, and 2E1 but decreased 12\u201339% following inhibition of drug\u2010metabolizing enzymes from CYP450 subfamilies 1A, 2C, 2D, and 3A. These results suggest that redundancy exists in the EXE metabolic pathway with multiple hepatic CYP450s catalyzing 17\u03b2\u2010DHE formation in\u00a0vitro. To further expand the knowledge of phase I EXE metabolism, the impact of CYP450 genetic variation on 17\u03b2\u2010DHE formation was assessed via enzyme kinetic parameters. Affinity for EXE substrate and enzyme catalytic velocity were calculated for hepatic wild\u2010type CYP450s and their common nonsynonymous variants by monitoring the reduction of EXE to 17\u03b2\u2010DHE. Several functional polymorphisms in xenobiotic\u2010metabolizing CYP450s 1A2, 2C8, 2C9, and 2D6 resulted in deviant enzymatic activity relative to wild\u2010type enzyme. Thus, it is possible that functional polymorphisms in EXE\u2010metabolizing CYP450s contribute to inter\u2010individual variability in patient outcomes by mediating overall exposure to the drug and its active metabolite, 17\u03b2\u2010DHE.Exemestane ( Overall, glucuronidation (Vmax/KM) of 17\u03b2\u2010DHE in the HLM panel was 36\u2010fold lower in UGT2B17\u2010null homozygotes (*2/*2) compared to wild\u2010type homozygotes (*1/*1) , such as the CYP450s, can lead to genotype\u2010dependent variability in clinical responses for certain drugs , Tokyo Chemical Industry Co. or Sigma\u2010Aldrich . Thin\u2010layer chromatography plates and silica columns used to purify the synthesized steroids were purchased from Bonna\u2010Agela Technologies Inc. and Yamazen Corp. , respectively. Variant CYP450 overexpression vectors were made using a QuikChange II Site\u2010Directed Mutagenesis Kit from Agilent , as well as oligonucleotides manufactured by Integrated DNA Technologies . Negative control baculosomes, SuperScript II First\u2010Strand Synthesis System for RT\u2010PCR, cell culture medium, fetal bovine serum, penicillin/streptomycin, and G418 were acquired from Invitrogen . Choice\u2010Taq polymerase was purchased from Denville Scientific . LC/MS grade organic solvents, tris base, glycine, tetramethylethylenediamine (TEMED), ammonium persulfate (APS), sodium dodecyl sulfate (SDS), SuperSignal West Femto Maximum Sensitivity Substrate, Lipofectamine 2000, oligo(dT) primer, Pierce BCA protein assay kit, PVDF membranes, and monoclonal HRP\u2010conjugated V5 epitope antibody were procured from Thermo Fisher Scientific . Nonfat dry milk used as a blocking agent for Western blotting was prepared by BioRad . The nicotinamide adenine dinucleotide phosphate (NADPH) regeneration system included in kinetic assays was purchased from Corning . Ampicillin, dithiothreitol (DTT), Tween 20, and acrylamide/bis\u2010acrylamide solution were acquired from Sigma\u2010Aldrich . Pooled mixed gender liver microsomes from 50 human donors were obtained from XenoTech . The ethnic composition of the pooled hepatic microsomes was 84% Caucasian, 8% Hispanic, 6% African American, and 2% Asian. (\u2010)\u2010N\u20103\u2010benzylphenobarbital (NBP) is a product of Cypex Ltd , whereas all other compounds used for CYP450 isoform\u2010specific inhibition were obtained from Sigma\u2010Aldrich.EXE and its phase I metabolites were synthesized at Washington State University using previously published protocols Variation Viewer . Search 20 was used to prime reverse transcription of pooled hepatic total RNA from five human donors (Penn State Cancer Institute Biorepository). The resultant first\u2010strand cDNA was further amplified using CYP450 isoform\u2010specific primers and Taq polymerase. cDNA encoding wild\u2010type CYP450s 1A2, 2C8, 2C9, 2C19, 2D6, 3A4, or 3A5 was introduced into the pcDNA3.1/V5\u2010His\u2010TOPO expression vector for stable overexpression in mammalian cells. CYP2D6*2 and CYP2C19*1B cDNA were also amplified from pooled human liver RNA. Additional constitutive overexpression vectors encoding nonsynonymous variants with MAF > 0.01 were produced for CYP450s 1A2, 2C8, 2C9, 2C19, and 3A4 via site\u2010directed mutagenesis (SDM) using wild\u2010type plasmid as template. Two common CYP2D6 haplotypes were likewise derived from the CYP2D6*2 overexpression vector through SDM. Oligonucleotide pairs used to amplify wild\u2010type CYP450 cDNA or prime mutagenesis are detailed in Data Oligo(dT)\u03bcg/mL G418 selective pressure in DMEM supplemented with 4.5\u00a0g/L glucose, l\u2010glutamine, 110\u00a0mg/L sodium pyruvate, 10% FBS, and penicillin/streptomycin. Data Each expression vector was transformed into chemically competent BL21. Transformants were then grown overnight on ampicillin selection plates at 37\u00b0C. Sanger sequencing was used for sequence confirmation prior to transfecting HEK293 with overexpression plasmids using Lipofectamine 2000 per the manufacturer instructions. Transfected HEK293 were grown for at least 3\u00a0weeks under 700\u00a0\u03bcL reaction contained 2.5\u00a0mmol/L EXE substrate, 3\u00a0\u03bcL NADPH regeneration system, and 20\u00a0\u03bcg of microsomal protein from a wild\u2010type CYP450\u2010overexpressing HEK293 cell line in phosphate\u2010buffered saline (PBS), pH 7.4. Similar incubations were conducted to assess background activity of endogenous metabolizing enzymes in non\u2010transfected HEK293, as well as commercially available negative control baculosomes. These incubations included 100\u00a0\u03bcM EXE, 50\u00a0\u03bcg of microsomes, or 50\u00a0\u03bcg of baculosomes in addition to PBS, pH7.4, and an NADPH regeneration system. Enzymatic incubations were terminated with 50\u00a0\u03bcL of ice\u2010cold acetonitrile before centrifugation at 13,200g for 15\u00a0min at 4\u00b0C. The resulting supernatant was analyzed by ultra\u2010pressure liquid chromatography paired with tandem mass spectrometry (UPLC/MS/MS) on a Waters ACQUITY platform configured with a 0.2\u00a0\u03bcm inline filter preceding a 1.7\u00a0\u03bcm ACQUITY UPLC BEH C18 column . The UPLC gradient used to separate EXE and its phase I metabolites has been described previously , 17\u03b1\u2010DHE (m/z 299.14 \u2192 135.07), 17\u03b2\u2010DHE (m/z 299.20 \u2192 135.13), and 6\u2010HME (m/z 312.89 \u2192 158.98). 0.01\u00a0sec dwell time was optimal for all four compounds, whereas 25\u00a0V of collision energy was used for EXE and 6\u2010HME. 20\u00a0V of collision energy was used for 17\u03b1\u2010 and 17\u03b2\u2010DHE. An additional non\u2010targeted screening for phase I EXE metabolites was performed using UPLC/MS to detect positively charged molecular ions with m/z ranging from 100 to 450. The identity of all metabolites observed was verified by comparing the observed retention times and m/z with those of purified standards.CYP450\u2010derived phase I EXE metabolites were identified in 45\u2010min incubations performed at 37\u00b0C. Each 50\u2010\u03b2\u2010DHE production was measured in 45\u2010min incubations at 37\u00b0C in PBS, pH 7.4 with varying concentrations of EXE (25\u20132500\u00a0\u03bcmol/L). Each reaction included 20\u00a0\u03bcg of microsomal protein from CYP450\u2010overexpressing HEK293, as well as an NADPH regeneration system. The chosen incubation length and protein concentration fell within the linear range of EXE reduction velocity curves for the seven wild\u2010type CYP450s assayed (data not shown). Cold acetonitrile (50\u00a0\u03bcl) was used to terminate each reaction. After centrifugation for 15\u00a0min at 4\u00b0C at 13,200g, 17\u03b2\u2010DHE formation was monitored in the supernatant according to a previously established UPLC/MS/MS method and quantitated against a standard curve constructed of known 17\u03b2\u2010DHE concentrations . Furafylline (1\u00a0\u03bcmol/L) was used to inhibit CYP1A2, whereas 0.5\u00a0\u03bcmol/L tranylcypromine (TCP) and 10\u00a0\u03bcmol/L thioTEPA inhibited CYPs 2A6 and 2B6, respectively. Other compounds used to systematically inhibit hepatic CYP450s included 0.5\u00a0\u03bcmol/L montelukast (CYP2C8 inhibitor), 1\u00a0\u03bcmol/L sulfaphenazole (CYP2C9 inhibitor), 0.5\u00a0\u03bcmol/L NBP (CYP2C19), 1\u00a0\u03bcmol/L quinidine (CYP2D6 inhibitor), 5\u00a0\u03bcmol/L clomethiazole (CYP2E1 inhibitor), and 1\u00a0\u03bcmol/L ketoconazole (CYP3A inhibitor). Initial dosages were selected from the literature and then further titrated to determine the lowest dose producing maximum isoform\u2010specific inhibition and analyzed by UPLC/MS/MS using the aforementioned method. A negative control reaction was run in parallel and received ethanol vehicle rather than inhibitor. Organic solvent constituted less than 1% of each incubation.Reduction of EXE to 17KM and relative Vmax values were calculated in GraphPad Prism 6 according to the Michaelis\u2013Menten equation\u00a0. Vmax is expressed as pmol\u00a0min\u22121\u00a0mg\u22121 and was normalized to total protein content to account for differences in CYP450 expression between cell lines as determined by Western blot analysis. Two\u2010sided unpaired t\u2010tests were used to compare the wild\u2010type catalytic activity of CYP450s 1A2 and 3A4 to their respective variants. Wild\u2010type CYP450s 2C8, 2C9, 2C19, and 2D6 were compared to their nonsynonymous variants using one\u2010way ANOVA supplemented with Dunnett's multiple comparison test. In all instances, the threshold for statistical significance was set at a two\u2010tailed P\u00a0<\u00a00.05. The percent change in 17\u03b2\u2010DHE formation associated with each inhibitor in pooled HLM was likewise calculated in GraphPad Prism 6. An uninhibited reaction receiving vehicle was considered maximum catalytic activity for the purposes of comparison. All experimental results represent triplicate assays performed independently.\u03b2\u2010DHE. Low levels of 6\u2010HME and 17\u03b1\u2010DHE formation were observed for all CYPs studied. In all cases, formation of the 17\u03b1\u2010dihydroexemestane (17\u03b1\u2010DHE) and 6\u2010hydroxymethylandrosta\u20101,4,6\u2010triene\u20103,17\u2010dione (6\u2010HME) metabolites exceeded the minimum threshold of detection. No EXE metabolite formation was observed in incubations using microsomes from the parent HEK293 cell line followed by CYP2C8 (KM\u00a0=\u00a00.66\u00a0\u00b1\u00a00.10\u00a0mmol/L), CYP3A5 (KM\u00a0=\u00a00.69\u00a0\u00b1 0.18\u00a0mmol/L), CYP1A2 (KM\u00a0=\u00a00.74\u00a0\u00b1\u00a00.5\u00a0mmol/L), and CYP3A4 (KM\u00a0=\u00a00.83\u00a0\u00b1\u00a00.16\u00a0mmol/L) . CYP1A2 catalyzed 17\u03b2\u2010DHE production at a rate of 61\u00a0\u00b1\u00a017\u00a0pmol\u00a0min\u22121\u00a0mg \u22121 followed by CYP2C19 (51\u00a0\u00b1\u00a08\u00a0pmol\u00a0min\u22121\u00a0mg \u22121) and CYP2D6 (49\u00a0\u00b1\u00a03\u00a0pmol\u00a0min\u22121\u00a0mg \u22121). Catalytic velocities for CYP450s 3A4 and 3A5 were not significantly different (39\u00a0\u00b1\u00a07 and 36\u00a0\u00b1\u00a04\u00a0pmol\u00a0min\u22121\u00a0mg \u22121). Intrinsic clearance (Vmax/KM) was calculated for each wild\u2010type enzyme as an indication of its overall catalytic activity against EXE. CYP2C8 exhibited the highest intrinsic clearance value (194\u00a0nL\u00a0min\u22121\u00a0mg\u22121) followed by CYP2D6 (86\u00a0nL\u00a0min\u22121\u00a0mg\u22121), CYP1A2 (82\u00a0nL\u00a0min\u22121\u00a0mg\u22121), CYP2C19 (55\u00a0nL\u00a0min\u22121\u00a0mg\u22121), CYP3A5 (52\u00a0nL\u00a0min\u22121 mg\u22121), and CYP3A4 (47\u00a0nL\u00a0min\u22121\u00a0mg\u22121). In addition to having the slowest rate of EXE reduction, CYP2C9 also exhibited the lowest overall catalytic activity against EXE substrate (27\u00a0nL\u00a0min\u22121\u00a0mg\u22121).Representative kinetic curves of 17In all, 23 nonsynonymous polymorphisms with MAF\u00a0>\u00a00.01 were detected using the NCBI Variation Viewer to search for common variants in hepatic CYP450s involved in xenobiotic metabolism. Four polymorphisms of interest were identified for CYP2C9. Two variants were gleaned for CYP2C19, whereas a single variant was reported for both CYP450s 1A2 and 3A4. No variants matching the search criteria were listed for CYP3A5. Variation Viewer search filters returned four common single nucleotide polymorphisms (SNPs) in CYP2C8. In the present study, two of the CYP2C8 polymorphisms were investigated as a single haplotype (CYP2C8*3). In all, 11 nonsynonymous polymorphisms occur in CYP2D6 at > 1%. Although CYP2D6 is notoriously polymorphic, only a handful of common haplotypes are associated with clinically relevant alterations in drug metabolism Zhou . For thiVmax was comparable between wild\u2010type CYP1A2 and CYP1A2Ser298Arg, the variant enzyme exhibited significantly increased affinity for EXE with KM values of 0.74\u00a0\u00b1\u00a00.5 and 0.59\u00a0\u00b1\u00a00.2\u00a0mmol/L, respectively. No notable differences were observed in affinity between wild\u2010type CYP450s 2C8, 2C9, 2C19, or 3A4 and their respective variants (Table\u00a0KM\u00a0=\u00a00.95\u00a0\u00b10.10\u00a0mmol/L) and CYP2D6*10 (KM\u00a0= 1.04\u00a0\u00b1\u00a00.01\u00a0mmol/L) were both associated with decreased affinity relative to wild\u2010type CYP2D6 (KM\u00a0=\u00a00.57 \u00b1\u00a00.03\u00a0mmol/L). The CYP2C8*2 and CYP2C8*4 polymorphisms were associated with 2.2 and 1.7\u2010fold increases in velocity of EXE reduction compared to wild\u2010type. CYP2C9*8 and CYP2C9*9 variants were likewise associated with increased rates of 17\u03b2\u2010DHE formation (Vmax\u00a0=\u00a073\u00a0\u00b1\u00a013 and 116\u00a0\u00b1\u00a013 versus 26\u00a0\u00b1\u00a00.6 pmol min\u22121\u00a0mg\u22121 for wild\u2010type). The Vmax for EXE reduction by CYP2D6*2 (23\u00a0\u00b1\u00a00.6\u00a0pmol\u00a0min\u22121\u00a0mg \u22121) is 53% lower than that of wild\u2010type CYP2D6 (49\u00a0\u00b1\u00a03 pmol\u00a0min\u22121\u00a0mg \u22121). Vmax values were comparable between CYP2C19 and its variants CYP2C19Ile331Val and CYP2C19Glu92Asp. No significant difference in Vmax was observed between wild\u2010type CYP3A4 and CYP3A4Arg162Gln .Although \u03b2\u2010DHE production . Similar results were obtained when either thioTEPA or clomethiazole were included to chemically inhibit CYP2B6 (112.5\u00a0\u00b1\u00a08.3% control activity) or CYP2E1 (97.3\u00a0\u00b1\u00a05.3% control activity), respectively. Inhibition of CYP2D6 decreased EXE reduction by 27%, whereas furafylline\u2010induced CYP1A2 inhibition reduced formation of the active 17\u03b2\u2010DHE metabolite by 19%.In incubations of EXE with pooled mixed gender HLMs, inhibition of CYP3A isoforms with ketoconazole resulted in a 39% decrease in 17ion Fig.\u00a0. Isoform\u03b2\u2010DHE represents a major metabolic pathway for this commonly prescribed aromatase inhibitor . CYPs 3A4 and 3A5 exhibited intermediate intrinsic clearance values in the present study , but the literature indicates that CYP3A isoforms are well expressed in the liver comprising 30% of total CYP450 content and accounting for nearly 55% of xenobiotic metabolism (Chang and Kam \u22121\u00a0mg\u22121) was second only to that of CYP2C8 (194\u00a0nL\u00a0min\u22121\u00a0mg\u22121) in kinetic assays. CYP2D6 is estimated to account for 30% of drug metabolism despite constituting only 2% of total hepatic CYP450 content (Chang and Kam INT\u00a0=\u00a082\u00a0nL\u00a0min\u22121\u00a0mg\u22121), accounts for only 2% of overall CYP450\u2010mediated xenobiotic metabolism in human liver in the presence of the isoform\u2010specific inhibitor ketoconazole suggests that CYP3As are the major hepatic CYP450s responsible for EXE reduction in human liver microsomes in the present study. CYP1A1 expression, in turn, is low in human liver diminishing the likelihood that it is a key participant in phase I EXE metabolism . Despite its high incidence in African populations (MAF\u00a0=\u00a00.0893), a three\u2010dimensional structure examining conformational changes in CYP1A2 arising from arginine substitution is currently unavailable , CYP2C8Arg139Lys,Lys399Arg (*3), and CYP2C8Ile264Met (*4) are comparable to wild\u2010type CYP2C8 in affinity for EXE substrate in\u00a0vitro. Kaspera et\u00a0al. and 6\u2010hydroxyl\u2010 (M\u201023) cerivastatin metabolites. Although E.\u00a0coli\u2010expressed CYP2C8*2 yielded individual Vmax values similar to wild\u2010type CYP2C8 for M\u20101 and M\u201023 formation, a 53% increase in the sum of cerivastatin metabolite clearance was noted . Kaspera et\u00a0al. and CYP2C9Ile359Leu (*3) were similar to those of wild\u2010type CYP2C9. The CYP2C9*2 and *3 polymorphisms are relatively common (MAF\u00a0>\u00a00.01) in South Asian, European, and Hispanic populations (see Data Arg150His (*8) and CYP2C9His251Arg (*9) polymorphic protein reduced EXE to 17\u03b2\u2010DHE 2.8\u2010 and 4.5\u2010fold faster than wild\u2010type CYP2C9, respectively. The CYP2C9*8 allele also increased clearance of the antidiabetic tolbutamide in\u00a0vitro relative to wild\u2010type CYP2D6 due to significant decreases in affinity and 17\u03b2\u2010DHE formation rate. Sakuyama et\u00a0al. increased the KM value for EXE substrate by 82% causing a 56% decrease in clearance compared to CYP2D6*1. CYP2D6*10 is highly prevalent in Asians and has previously been associated with decreased catalytic activity in\u00a0vitro is common in individuals of African heritage and is likewise considered a reduced function allele and production of the major active metabolite 17\u03b2\u2010DHE (Vmax) indicate that EXE metabolism by common allelic variants of CYP2C19 and CYP3A4 is comparable to that of their respective wild\u2010type CYP450s. The neutral effect of the CYP2C19Ile331Val (*1B) polymorphism on EXE reduction confirms prior observations of equivalent catalytic activity for the CYP2C19*1A and *1B alleles did not directly impact 17\u03b2\u2010DHE formation, it cosegregates with approximately 20% of CYP2C19*2 poor metabolizing alleles in Caucasians polymorphism does not appreciably impact EXE metabolism. However, kinetic studies of the variant enzyme with additional substrates are needed to confirm its overall effect on CYP3A4 activity.Kinetic parameters measuring substrate affinity (\u03b1\u2010DHE, as well as the active metabolite 17\u03b2\u2010DHE. Earlier studies suggested that 17\u03b2\u2010DHE may partially determine overall drug exposure by acting as an androgen agonist, contributing to estrogen blockade through aromatase inhibition, and by serving as a gateway to phase II conjugation and excretion (Ariazi et\u00a0al. \u03b2\u2010DHE formation or clearance may contribute to inter\u2010individual variation in the overall therapeutic efficacy of EXE by altering a major metabolic pathway. This possibility is bolstered by the observation that three of the variant CYP450s included in this study had altered affinity for EXE substrate leading to differential 17\u03b2\u2010DHE production while five variants had deviant catalytic rates of EXE reduction. To our knowledge, this is the first study to report the impact of common nonsynonymous polymorphisms in CYP450s on EXE reduction to its 17\u03b2\u2010DHE metabolite. A previous in\u00a0vitro study demonstrated the capacity of genetic variation to alter EXE metabolism by the cytosolic ketosteroid reductases CBR1 and AKR1C1\u20104 (Platt et\u00a0al. The present in\u00a0vitro study augments existing pharmaceutical knowledge by examining the role of hepatic CYP450s in the metabolism of EXE, a widely used endocrine therapy for hormone\u2010responsive breast cancer. Qualitative enzymatic incubations with EXE confirm that multiple hepatic monoxygenases from CYP450 families 1, 2, and 3 catalyze the production of 6\u2010HME, 17There are no conflicts of interest.Data S1. Digital Content 1.doc.Click here for additional data file.Data S2. Digital Content 2.doc.Click here for additional data file.Data S3. Digital Content 3.doc.Click here for additional data file.Data S4. Digital Content 4.doc.Click here for additional data file."} +{"text": "The title crystal is a co-crystal with the 1,8-naphthyridine derivative crystallizing with one mol\u00adecule of succinimide per formula unit. In the crystal, the two mol\u00adecules are mutually linked by N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds. The packing is consolidated by C\u2014H\u22ef hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions. 17H13Br2N3O\u00b7C4H5NO2, is a co-crystal of N-benzamide and pyrrolidine-2,5-dione (succinimide). The benzamide mol\u00adecule exhibits pseudo-mirror symmetry, with an r.m.s. deviation of the non-H atoms of 0.09\u2005\u00c5 (except for the two Br atoms). The angle between the least-squares planes of the two mol\u00adecules is 26.2\u2005(2)\u00b0. In the crystal, the two mol\u00adecules are mutually linked by N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds. The packing is consolidated by C\u2014H\u22ef hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions.The title compound, C The naphthyridine ring system makes a dihedral angle of 2.2\u2005(2)\u00b0 with the benzene ring and is oriented at an angle of 26.2\u2005(2)\u00b0 relative to the succinimide. The conformation of the C=O and the N\u2014H bonds of the amide segment are anti to one another, similar to that reported for benzamide moiety in N-{4-[(6-chloro\u00adpyridin-3-yl)-meth\u00adoxy]phen\u00adyl}-2,6-di\u00adfluoro\u00adbenzamide \u2005\u00c5 , forming a three-dimensional supra\u00admolecular network N\u2014H\u22efN bond bifurcated Table\u00a01. In the rk Fig.\u00a03.et al., 2016et al., 2013In the Cambridge Structural Database benzamide were added to an dry aceto\u00adnitrile (30\u2005ml) solution under nitro\u00adgen atmosphere. The mixture was refluxed at room temperature in the presence of light with a 250\u2005W infrared lamp for 4\u2005h. Excess solvent was removed and the crude product was purified by column chromatography using di\u00adchloro\u00admethane/methanol (120:1) as the mobile phase to give a light-yellow powder . Crystals suitable for X-ray analysis were obtained by slow diffusion of a di\u00adchloro\u00admethane solution at ambient temperature. Several cycles of purification by chromatography were used to reduce the amount of succinimide.Uiso(H) = 1.5Ueq(C) for methyl H atoms and Uiso(H) = 1.2Ueq(C) for all other H atoms, and with N\u2014H = 0.86\u2005\u00c5, Uiso(H) = 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016019034/wm5334sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016019034/wm5334Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016019034/wm5334Isup3.cmlSupporting information file. DOI: 1519551CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atoms within N2S2 donor sets because of the presence of chelating thio\u00adsemicarbazone anions. Supra\u00admolecular chains are found in the crystal owing to the formation of thio\u00adamide-N\u2014H\u22efS(thione) hydrogen bonds and eight-membered thio\u00adamide {\u22efHNCS}2 synthons.The title thio\u00adsemicarbazone compound features tetra\u00adhedrally coordinated Zn II complex, [Zn(C19H20N3OS)2] {systematic name: bis\u00ad[(N-ethyl-N\u2032-{(Z)-[(2E)-3-(4-meth\u00adoxy\u00adphen\u00adyl)-1-phenyl\u00adprop-2-en-1-yl\u00adidene]amino}\u00adcarb\u00adam\u00adim\u00adid\u00ado\u00adyl)sulfanido]zinc(II)}, features a tetra\u00adhedrally coordinated ZnII ion within an N2S2 donor set provided by two N,S-chelating thio\u00adsemicarbazone anions. The resulting five-membered Zn,C,N2,S chelate rings adopt different conformations, i.e. almost planar and an envelope with the Zn atom being the flap atom. The configuration about the imine bond within the chelate ring is Z but those about the exocyclic imine and ethyl\u00adene bonds are E. In the crystal, supra\u00admolecular [100] chains mediated by thio\u00adamide-N\u2014H\u22efS(thione) hydrogen bonds and eight-membered thio\u00adamide {\u22efHNCS}2 synthons are observed. A range of inter\u00adactions, including C\u2014H\u22efO, C\u2014H\u22ef\u03c0, C\u2014H\u22ef\u03c0(chelate ring) and \u03c0(meth\u00adoxy\u00adbenzene)\u2014\u03c0(chelate ring) consolidate the packing. The Hirshfeld surface analysis performed on the title complex also indicates the influence of the inter\u00adactions involving the chelate rings upon the packing along with the more conventional contacts.The title Zn R1R2C=N\u2014N(R3)\u2014C(=S)NR4R5 for R1\u20135 = H/alk\u00adyl/aryl, are numerous and multi-functional. Their preparation is often facile, being formed from the condensation reaction between an aldehyde (or a ketone) with the amine group of a thio\u00adsemicarbazide precursor. In the same way, the diversity in ligand construction ensures a rich coord\u00adination chemistry \u00b0 for the S1\u2014Zn\u2014N3 chelate angle to 127.92\u2005(4)\u00b0 for S1\u2014Zn\u2014S2. The assignment of four-coordinate geometries can be qu\u00adanti\u00adfied by comparing the calculated value of \u03c44, in this case 0.74, with the ideal values for an ideal tetra\u00adhedron, i.e. 1.00, and perfect square-planar geometry, i.e. 0.00 2 chelate rings, and these adopt different conformations. Whereas, the ring is almost planar (r.m.s. deviation = 0.0325\u2005\u00c5), the chelate ring is best described as an envelope with the Zn atom lying 0.205\u2005(5)\u2005\u00c5 out of the plane of the remaining four atoms (r.m.s. deviation = 0.0011\u2005\u00c5). The dihedral angle between the mean planes through the chelate rings is 79.68\u2005(8)\u00b0. To a first approximation, the thio\u00adsemicarbazone ligands comprise two planar regions. Thus, the non-hydrogen, non-phenyl atoms of the atoms of the S1-ligand define one plane (r.m.s. deviation = 0.1910\u2005\u00c5), which forms a dihedral angle of 54.53\u2005(8)\u00b0 with the (C14\u2013C19) ring, consistent with a near perpendicular relationship. The comparable values for the S2-ligand are 0.2800\u2005\u00c5 and 75.09\u2005(11)\u00b0, respectively.The mode of the coordination of the thio\u00adsemicarbazone ligands leads to the formation of five-membered ZnSCNc-axis direction sustained by eight-membered thio\u00adamide {\u22efHNCS}2 synthons, Fig.\u00a02a and Table\u00a02b, it is evident that two rows of mol\u00adecules, each with a right-angle topology, are connected by N\u2014H\u22efS(thione) hydrogen bonds. Centrosymmetrically related right angles are connected into a supra\u00admolecular tube, Fig.\u00a02c, via imine-phenyl-C\u2014H\u22efO(meth\u00adoxy), imine-phenyl-C\u2014H\u22ef\u03c0(imine-phen\u00adyl) and imine-phenyl-C\u2014H\u22ef\u03c0(meth\u00adoxy\u00adbenzene) inter\u00adactions, Table\u00a02et al., 2014et al., 2016v with a ring centroid\u2013ring centroid separation of 3.778\u2005(2)\u2005\u00c5 and angle of inclination = 15.04\u2005(17)\u00b0 for symmetry operation (v): 2\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z. A review has appeared very recently on the topic of \u03c0(chelate ring)\u2013\u03c0(arene) and \u03c0(chelate ring)\u2013\u03c0(chelate ring) inter\u00adactions where it was suggested that inter\u00adactions of the former type provide comparable energies of stabilization to mol\u00adecular packing as do weak conventional hydrogen bonds ring and between ethyl\u00adene-C24\u2014H and the ring, Table\u00a02a. The donors and acceptors of the inter\u00admolecular N\u2014H\u22efS and C\u2014H\u22efO inter\u00adactions are also viewed as blue and red regions about respective atoms in the images of Fig.\u00a04a and b, respectively.The Hirshfeld surfaces calculated for (I)a, and those delineated into H\u22efH, C\u22efH/H\u22efC, S\u22efH/H\u22efS and O\u22efH/H\u22efO contacts de + di \u223c\u20052.1\u2005\u00c5 in Fig.\u00a06a is due to a short inter\u00adatomic H\u22efH contact are also reflected from the small but important contribution from C\u22efN/N\u22efC and C\u22efS/S\u22efC contacts, Table\u00a04The overall two dimensional fingerprint plot for (I)t Table\u00a03 and the N\u2032-{(E)--amino}-N-ethyl\u00adcarbamimido\u00adthio\u00adato-\u03ba2N\u2032,S)zinc(II) mol\u00adecule, which differs from (I)et al., 20172S2 coordination geometries are found with values of \u03c44 of 0.70 and 0.74 for the two independent mol\u00adecules comprising the asymmetric unit. Indeed, in the publication describing this structure =NN=CR\u2032R\u2032\u2032]2 and all structures adopt the same basic structural motif as described herein for (I)The most relevant structure available for comparison is that of the recently described bis\u00ad was dissolved in hot absolute ethanol (30\u2005ml), which was added to a solution of Zn(CH3COO)2\u00b72H2O in hot absolute ethanol (40\u2005ml). The mixture was heated and stirred for about 10\u2005min, followed by stirring for 1\u2005h at room temperature. The yellow precipitate obtained was filtered, washed with cold ethanol and dried in vacuo. Single crystals were grown at room temperature from the slow evaporation of the title compound in a mixed solvent system containing di\u00admethyl\u00adformamide and aceto\u00adnitrile . IR (cm\u22121): 3351 \u03bd(N\u2014H), 1597 \u03bd(C=N), 1009 \u03bd(N\u2014N), 420 \u03bd(M\u2014N), 362 \u03bd(M\u2014S).Analytical grade reagents were used as procured and without further purification. 4-Ethyl-3-thio\u00adsemicarbazide and 4-meth\u00adoxy\u00adchalcone were dissolved separately in hot absolute ethanol (30\u2005ml) and mixed while stirring. About five drops of concentrated hydro\u00adchloric acid were added to the mixture to catalyse the reaction. The reaction mixture was heated and stirred for about 20\u2005min, and stirring was continued for another 30\u2005min at room temperature. The resulting yellow precipitate, 4-meth\u00adoxy\u00adchalcone-4-ethyl-3-thio\u00adsemicarbazone, was filtered off, washed with cold absolute ethanol and dried Uiso(H) set to 1.2\u20131.5Ueq(C). The nitro\u00adgen-bound H atoms were located in a difference-Fourier map but were refined with a distance restraint of N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.2Ueq(N). The maximum and minimum residual electron density peaks of 1.10 and 0.59\u2005e\u2005\u00c5\u22123, respectively, are located 1.04 and 0.71\u2005\u00c5 from the Zn atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989018000282/hb7725sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018000282/hb7725Isup2.hklStructure factors: contains datablock(s) I. DOI: 1814817CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II ion is coordinated by two pyridine N atoms from two symmetry-related unsymmetrical dipyridyl ligands and two chloride anions in a highly distorted tetra\u00adhedral geometry. Each unsymmetrical dipyridyl ligand links two HgII ions into polymeric zigzag chains. In the crystal, the chains are linked into a three-dimensional supra\u00admolecular network by inter\u00admolecular N/C\u2014H\u22efCl hydrogen bonds and weak C\u2014H\u22ef\u03c0 inter\u00adactions. Weak C\u2014Cl\u22ef\u03c0 inter\u00adactions and Cl\u22efCl contacts between the network and the solvent chloro\u00adform mol\u00adecules are also observed.In the title compound, each Hg LCl2]\u00b70.5CHCl3}n (L = N-(pyridin-4-ylmeth\u00adyl)pyridin-3-amine, C11H11N3), contains one HgII ion, one bridging L ligand, two chloride ligands and a chloro\u00adform solvent mol\u00adecule with half-occupancy that is disordered about a crystallographic twofold rotation axis. Each HgII ion is coordinated by two pyridine N atoms from two symmetry-related L ligands and two chloride anions in a highly distorted tetra\u00adhedral geometry with bond angles falling in the range 99.05\u2005(17)\u2013142.96\u2005(7)\u00b0. Each L ligand bridges two HgII ions, forming polymeric zigzag chains propagating in [010]. In the crystal, the chains are linked by inter\u00admolecular N/C\u2014H\u22efCl hydrogen bonds together with weak C\u2014H\u22ef\u03c0 inter\u00adactions, resulting in the formation of a three-dimensional supra\u00admolecular network, which is further stabilized by C\u2014Cl\u22ef\u03c0 inter\u00adactions between the solvent chloro\u00adform mol\u00adecules and the pyridine rings of L [chloride-to-centroid distances = 3.442\u2005(11) and 3.626\u2005(13)\u2005\u00c5]. In addition, weak Cl\u22efCl contacts [3.320\u2005(5)\u2005\u00c5] between the chloro\u00adform solvent mol\u00adecules and the coordinating chloride anions are also observed.The asymmetric unit of the title compound, {[Hg The solvent mol\u00adecule is disordered over two orientations of equal occupancy about the crystallographic twofold rotation axis. As shown in Fig.\u00a01II ion is highly distorted tetra\u00adhedral with two coordination sites being occupied by two pyridine N atoms from two symmetry-related L ligands. The geometry of the HgII ion is completed by the coordination of two chloride ions. The tetra\u00adhedral angles around the HgII ion fall in the range of 99.05\u2005(17)\u2013142.96\u2005(7)\u00b0 \u2005\u00c5. In the L ligand, the Cpy\u2014N\u2014C\u2014Cpy torsion angle is \u221270.9\u2005(7)\u00b0 while the dihedral angle between two terminal pyridine ring planes is 85.0\u2005(2)\u00b0. The conformation of the L ligand, along with the the Npy\u2014Hg\u2014Npy coordination angle [99.05\u2005(17)\u00b0], may induce the zigzag topology of the chain.Each is Fig.\u00a02. The sepbc plane \u2005\u00c5, C12\u2014Cl4\u22efCg2 = 170.7\u2005(8)\u00b0, Cl5\u22efCg2iv = 3.626\u2005(13)\u2005\u00c5 and C12\u2014Cl5\u22efCg2iv 144.1\u2005(8)\u00b0 are observed.In the crystal, adjacent zigzag chains are linked by inter\u00admolecular N\u2014H\u22efCl hydrogen bonds and weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions Table\u00a02, forminge Figs. 2 and 3 \u25b8.L ligand was synthesized according to a literature method = 0.93\u2005\u00c5 for Csp2\u2014H, 0.97\u2005\u00c5 for methyl\u00adene C\u2014H, 0.98\u2005\u00c5 for methine C\u2014H, and 0.86\u2005\u00c5 for amine N\u2014H, and were refined as riding with Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016015310/sj5509sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989016015310/sj5509Isup2.hklStructure factors: contains datablock(s) I. DOI: 1507232CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "These findings provide additional understanding of the NBC series and will aid in the development of these NLRP3 inhibitors as tool compounds or therapeutic candidates for sterile inflammatory diseases.The NLRP3 inflammasome is an important regulator of the sterile inflammatory response, and its activation by host\u2010derived sterile molecules leads to the intracellular activation of caspase\u20101, processing of the pro\u2010inflammatory cytokines interleukin\u20101\u03b2 (IL\u20101\u03b2)/IL\u201018, and pyroptotic cell death. Inappropriate activation of NLRP3 drives a chronic inflammatory response and is implicated in several non\u2010communicable diseases, including gout, atherosclerosis, type\u2005II diabetes and Alzheimer's disease. In this study, we report the design, synthesis and biological evaluation of novel boron compounds (NBCs) as NLRP3 inflammasome inhibitors. Structure\u2013activity relationships (SAR) show that 4\u2010fluoro substituents on the phenyl rings retain NLRP3 inhibitory activity, whereas more steric and lipophilic substituents diminish activity. Loss of inhibitory activity is also observed if the CCl Sterile inflammation is a host\u2010driven immune response to injury in the absence of infection.NLRP3 gene causes spontaneous IL\u20101\u03b2 release in patients with cryopyrin\u2010associated periodic syndrome (CAPS) diseases that are characterised by fever, rashes and extensive joint pain.IL\u20101\u03b2 and NLRP3 activation are well characterised in a number of non\u2010communicable diseases involving sterile inflammation including gout,Given the critical role of NLRP3 and IL\u20101\u03b2 in human disease,A number of small molecule inhibitors of the NLRP3 inflammasome have been previously described.1), BC23 (2) and NBC6 group. However, the impact of phenyl ring substitutions on IL\u20101\u03b2 release was not assessed. Additionally, the presence of the CCl3 group significantly contributes to the high lipophilicity of these oxazaborine inhibitors, limiting their drug\u2010likeness.We recently reported on the discovery of new boron\u2010based small molecules as potent NLRP3 inhibitors.2) and NBC6 (3), in addition to seeking alternative bioisosteres of the CCl3 group in the search for new NLRP3 inhibitors with improved potency and drug\u2010like properties.Here we explored structure\u2013activity relationships (SAR) of phenyl ring substitutions based on the known NLRP3 inhibitors BC23 with magnesium turnings or isopropylmagnesium chloride (iPrMgCl), followed by treatment with one molar equivalent of trimethyl borate (B(OMe)3) to afford symmetrical borinic acids (5).4 (when X=MgBr), such as p\u2010tolylmagnesium bromide, with a stoichiometric quantity of phenylboronic acid pinacol ester (7) to give the asymmetric borinic acid (6).Using the Topliss scheme for aromatic substituents,5 and 6 were then reacted with either (Z)\u20102\u2010acetyl\u20103\u2010amino\u20104,4,4\u2010trichlorobut\u20102\u2010enamide or (Z)\u20104\u2010amino\u20105,5,5\u2010trichloropent\u20103\u2010en\u20102\u2010oneThe synthesised borinic acids m,AN0128, a known borinic acid picolinate ester prepared by Anacor Pharmaceuticals, was also synthesised in a two\u2010step method according to the reported procedure and NBC6 (3) were then designed where the CCl3 group was replaced with the bioisosteric trifluoromethyl (CF3) group to give analogues NBC41 and NBC42 to prime the cells and induce expression of pro\u2010IL\u20101\u03b2. The primed cells were then incubated with vehicle (0.5\u2009% DMSO) or inhibitor (10\u2005\u03bcm) for 15 minutes before the NLRP3 inflammasome activator nigericin was added to cells. Supernatants were removed and IL\u20101\u03b2 levels were analysed by ELISA. Nigericin induced release of IL\u20101\u03b2 and this was inhibited by our parent molecule BC23 as expected had minimal effect on inhibitory activity and 4\u2010CH3 (NBC38) derivatives all inhibited IL\u20101\u03b2 release to a similar extent. This observation is further supported by the significantly decreased activities of 3,4\u2010Cl2 (NBC39) and 3\u2010Cl,4\u2010Me (NBC40) derivatives which possess additional substitutions around the phenyl rings. In contrast, there was an observed correlation between increasing lipophilicity and decreasing inhibitory activity across the BC23 series (NBC35 < NBC36 \u2248NBC37 \u2248NBC38 < NBC39). These observations were also in agreement across the NBC6 series, where the disubstituted\u2010p\u2010tolyl analogue NBC32 had slightly lower IL\u20101\u03b2 inhibitory activity than the monosubstituted\u2010p\u2010tolyl analogue NBC33 and parent compound NBC6 in THP\u20101 cells . These results suggest that aryl ring substitution, particularly with more steric and lipophilic substituents, is unlikely to enhance the activity of the NBCs.We initially tested the effect of modifying the phenyl rings of NBCs on their ability to inhibit NLRP3 inflammasome\u2010dependent release of IL\u20101\u03b2 from macrophages. Immortalised bone marrow derived macrophages (iBMDMs) were treated with LPS (1\u2005\u03bcg\u2009mLIt was surprising to note the lack of inhibitory activity of AN0128 in the IL\u20101\u03b2 release assay given its known anti\u2010inflammatory effects against IL\u20101\u03b2 and TNF\u2010\u03b1.3 group had been replaced with the bioisosteric trifluoromethyl (CF3) group and NBC42 were ineffective under these conditions . Therefore these initial studies suggest that the CCl3 group is not labile under these reaction conditions. It was noted that during these experiments, nucleophiles were found to attack the boron atom and undergo decomplexation. For example, BC23 cleanly hydrolyses into diphenylborinic acid and (Z)\u20104\u2010amino\u20105,5,5\u2010trichloropent\u20103\u2010en\u20102\u2010one in 9:1 [D6]DMSO/D2O solvent at 37\u2009\u00b0C with a half\u2010life of \u224824\u2005h, as monitored by 1H\u2005NMR spectroscopy .Given that the CClThe NLRP3 inflammasome is a multi\u2010molecular protein complex that is critical for inflammatory responses. Its formation leads to activation of caspase\u20101, which cleaves and activates IL\u20101\u03b2.3 group.3 group is key to inhibitory activity. These discoveries provide new insight into the activity of the NBC series and will aid future development of the NBC molecules as inflammasome inhibitors.We recently published the NBC series of NLRP3 inflammasome inhibitors reporting that key features required for bioactivity were the oxazaborine ring and CClGeneral: (Z)\u20104\u2010Amino\u20105,5,5\u2010trichloropent\u20103\u2010en\u20102\u2010one and (Z)\u20102\u2010acetyl\u20103\u2010amino\u20104,4,4\u2010trichlorobut\u20102\u2010enamide intermediates were prepared as previously described.1H, 13C, 11B and 19F\u2005NMR spectra were recorded on a Bruker Avance 400 or 300\u2005MHz spectrometer. Chemical shifts (\u03b4) are defined in parts per million (ppm). 1H\u2005NMR spectra were referenced to tetramethylsilane or residual undeuterated solvent . 13C\u2005NMR spectra were referenced to residual undeuterated solvent as an internal reference. 11B\u2005NMR chemical shifts were referenced to external reference BF3\u22c5OEt2 (\u03b4=0.0\u2005ppm). 19F\u2005NMR chemical shifts were referenced using the deuterium lock signal of the solvent. ESI and APCI mass spectrometry was carried out on a Waters Acquity UPLC system connected to a Waters SQD2 mass spectrometer. Accurate mass determination was carried out on a Thermo Exactive Plus EMR Orbitrap LC\u2013MS system. Molecular ion peaks are defined as mass/charge (m/z) ratios. Infrared spectroscopy was recorded on a JASCO FT/IR\u20104100 spectrophotometer using the Spectra Manager\u2005II (JASCO) software package. Melting points were measuring using a Stuart SMP10 melting point apparatus. Lyophilisation was carried out using a Christ alpha1\u20104 plus freeze dryer equipped with an Edwards vacuum pump. Microwave irradiation was carried out on a Biotage Initiator Classic microwave using 2\u20135\u2005mL Biotage glass vials. Analytical thin\u2010layer chromatography (TLC) was performed using silica gel 60 on aluminium sheets coated with F254 indicator. All spots were visualised with KMnO4 or ultraviolet light using a MV Mineralight lamp (254/365) UVGL\u201058. Flash column chromatography was performed using silica gel with particle size 40\u201363\u2005\u03bcm. Evaporation of solvents was conducted on a B\u00fcchi Rotavapor R\u2010200.p\u2010tolylborinic acid (5\u2009a)Di\u2010: To an oven\u2010dried Schlenk flask under N2 was added p\u2010tolylmagnesium bromide in anhydrous THF (5\u2005mL). B(OMe)3 was added dropwise to the reaction mixture and stirred at room temperature for 3\u2005h. HCl was then added and stirred for 30\u2005min to quench the reaction. The reaction mixture was extracted with EtOAc (3\u00d710\u2005mL), washed with brine (1\u00d710\u2005mL), dried over MgSO4, filtered and evaporated in\u2005vacuo. The crude product was then purified by flash column chromatography to give 5\u2009a as a colourless oil . 1H\u2005NMR : \u03b4=7.63 ), 7.18 ), 5.65 , 2.33\u2005ppm ; 13C\u2005NMR : \u03b4=141.2 (B\u2010Ar(p)), 134.8 (B\u2010Ar(o)), 128.7 (B\u2010Ar(m)), 21.7\u2005ppm (CH3), B\u2010Ar(i) quaternary signal not observed.p\u2010tolyl)\u20104\u2010(trichloromethyl)\u20102,3\u2010dihydro\u20101,3,2\u2010oxazaborinin\u20101\u2010ium\u20102\u2010uide (NBC32)5\u2010Acetyl\u20106\u2010amino\u20102,2\u2010bis(: (Z)\u20102\u2010Acetyl\u20103\u2010amino\u20104,4,4\u2010trichlorobut\u20102\u2010enamide was added to a solution of 5\u2009a in anhydrous THF (5\u2005mL). The mixture was stirred at 50\u2009\u00b0C under N2 for 16\u2005h. The mixture was concentrated in\u2005vacuo and purified by flash column chromatography . The collected fractions were combined, evaporated in\u2005vacuo and stirred in cold n\u2010hexane (15\u2005mL) for 30\u2005min. The precipitate was then filtered and dried under vacuum to give NBC32 as a yellow solid ; mp: 145\u2013148\u2009\u00b0C (dec); 1H\u2005NMR : \u03b4=9.23 , 7.58 C=C), 7.29 ), 7.12 ), 5.92 , 2.34 , 2.32\u2005ppm ; 13C\u2005NMR : \u03b4=198.1 (CH3CO), 169.2 (CONH2), 165.5 (Cl3C(NH)C=C), 156.1 (B\u2010Ar(i)), 136.2 (B\u2010Ar(p)), 132.0 (B\u2010Ar(o)), 128.2 (B\u2010Ar(m)), 97.9 (Cl3C(NH)C=C), 94.5 (CCl3), 34.0 (CH3CO), 21.3\u2005ppm (CH3); IR (neat): \u03bd\u02dc=3388 (N\u2212H), 3313 (N\u2212H), 1644 (C=O), 1608 , 1561\u2005cm\u22121 (C=C\u2010NH); MS(ES\u2212) (m/z): 434.2 \u2212, 435.2 \u2212, 436.2 \u2212, 437.2 \u2212, 438.2 \u2212, 439.2 \u2212; MS(ES+) (m/z): 344.1 +, 345.0 +, 346.0 +, 347.0 +, 349.1 +, 439.1 +; HRMS(ES+) (m/z): [M+H]+ calcd for C20H2111B35Cl3N2O2, 437.0756; found, 437.0738, error: 4.1\u2005ppm.p\u2010tolyl)borinic acid (6\u2009a)(Phenyl) in anhydrous THF (5\u2005mL). Phenylboronic acid pinacol ester in anhydrous THF (5\u2005mL) was added dropwise to the reaction mixture and stirred at room temperature for 3\u2005h. HCl was then added and stirred for 30\u2005min to quench the reaction. The reaction mixture was extracted with EtOAc (3\u00d710\u2005mL), washed with brine (1\u00d710\u2005mL), dried over MgSO4, filtered and evaporated in\u2005vacuo. The crude product was then purified by flash column chromatography to give 6\u2009a as a colourless oil . 1H\u2005NMR : \u03b4=7.72 ), 7.64 ), 7.32\u20137.47 ), 7.19 ), 5.74 , 2.33\u2005ppm ; 13C\u2005NMR : \u03b4=141.4 (B\u2010Ar(p)), 135.0 (B\u2010Ph(o)), 134.6 (B\u2010Ar(o)), 130.9 (B\u2010Ph(p)), 128.8 (B\u2010Ar(m)), 127.9 (B\u2010Ph(m)), 21.7\u2005ppm (CH3), B\u2010Ar(i) and B\u2010Ph(i) quaternary signals not observed; MS(ES\u2212) (m/z): 195.1 \u2212; HRMS(ES\u2212) (m/z): [M\u2212H]\u2212 calcd for C13H1211BO, 195.0987; found, 195.0973, error: 7.2\u2005ppm.p\u2010tolyl)\u20104\u2010(trichloromethyl)\u20102,3\u2010dihydro\u20101,3,2\u2010oxazaborinin\u20101\u2010ium\u20102\u2010uide (NBC33)5\u2010Acetyl\u20106\u2010amino\u20102\u2010(phenyl)\u20102\u2010(: (Z)\u20102\u2010Acetyl\u20103\u2010amino\u20104,4,4\u2010trichlorobut\u20102\u2010enamide was added to a solution of 6\u2009a in anhydrous THF (5\u2005mL). The mixture was stirred at 50\u2009\u00b0C under N2 for 16\u2005h. The mixture was concentrated in\u2005vacuo and purified by flash column chromatography . The collected fractions were combined, evaporated in\u2005vacuo and stirred in cold n\u2010hexane (15\u2005mL) for 30\u2005min. The precipitate was then filtered and dried under vacuum to give NBC33 as a yellow solid ; mp: 141\u2013143\u2009\u00b0C; 1H\u2005NMR : \u03b4=9.25 , 7.59 C=C), 7.40 ), 7.23\u20137.34 & B\u2010Ar(o)), 7.12 ), 5.94 , 2.34 , 2.31\u2005ppm ; 13C\u2005NMR : \u03b4=192.8 (CH3CO), 163.9 (Cl3C(NH)C=C), 160.3 (CONH2), 131.1 (B\u2010Ar(p)), 126.8 (B\u2010Ph(o)), 126.6 (B\u2010Ar(o)), 123.0 (B\u2010Ar(m)), 122.1 (B\u2010Ph(m)), 121.5 (B\u2010Ph(p)), 92.7 (Cl3C(NH)C=C), 89.2 (CCl3), 28.7 (CH3CO), 16.0\u2005ppm (CH3), B\u2010Ar(i) and B\u2010Ph(i) quaternary signals not observed; MS(ES\u2212) (m/z): 420.2 \u2212, 421.1 \u2212, 422.2 \u2212, 423.2 \u2212, 424.1 \u2212, 425.1 \u2212; HRMS(ES+) (m/z): [M+H]+ calcd for C19H1911B35Cl3N2O2, 423.0600; found, 423.0605, error: 1.2\u2005ppm.General procedure for synthesis of 2,2\u2010bisaryl\u20101,3,2\u2010oxazaborines: Using an adapted procedure,2 was added magnesium turnings (2.2\u2005equiv), anhydrous THF (5\u2005mL) and a small crystal of I2. The reaction was stirred at 40\u2009\u00b0C for 30\u2005min until complete decolourisation. A solution of aryl bromide (2\u2005equiv) and B(OMe)3 (1\u2005equiv) in anhydrous THF (5\u2005mL) was then added dropwise to the reaction mixture and then stirred for an additional 3\u2005h at 40\u2009\u00b0C. After cooling to room temperature, 1\u2009m HCl (10\u2005mL) was added and stirred for 30\u2005min to quench the reaction. The reaction mixture was extracted with EtOAc (3\u00d710\u2005mL), washed with brine (1\u00d710\u2005mL), dried over MgSO4, filtered and evaporated in\u2005vacuo to give the corresponding crude bisarylborinic acid, typically as a solid. (Z)\u20104\u2010Amino\u20105,5,5\u2010trichloropent\u20103\u2010en\u20102\u2010one (1.5\u2005equiv) was then added to crude bisarylborinic acid (1\u2005equiv) in anhydrous THF (5\u2005mL). The reaction was stirred at 50\u2009\u00b0C for 16\u2005h under N2. The reaction mixture was concentrated and purified by flash column chromatography using the indicated solvent system. Collected fractions were evaporated in\u2005vacuo and stirred in the minimum amount of cold n\u2010hexane for 30\u2005min to induce precipitation. The precipitate was then filtered and dried under vacuum to give the corresponding oxazaborine product. Percentage yields are reported over two steps.2,2\u2010Bis(4\u2010fluorophenyl)\u20106\u2010methyl\u20104\u2010(trichloromethyl)\u20102,3\u2010dihydro\u20101,3,2\u2010oxazaborinin\u20101\u2010ium\u20102\u2010uide (NBC35): EtOAc/n\u2010hexane, 1:9. Reaction scale: B(OMe)3 , 1\u2010bromo\u20104\u2010fluorobenzene and magnesium turnings gives bis(4\u2010fluorophenyl)borinic acid . Bis(4\u2010fluorophenyl)borinic acid and (Z)\u20104\u2010amino\u20105,5,5\u2010trichloropent\u20103\u2010en\u20102\u2010one gives NBC35 as a yellow solid ; mp: 130\u2013131\u2009\u00b0C; 1H\u2005NMR : \u03b4=7.31 ), 7.00 ), 5.83 C=CH), 2.25\u2005ppm , NH signal is overlapping with triplet at 7.31\u2005ppm; 13C\u2005NMR : \u03b4=186.5 (CH3CO), 166.1 (Cl3C(NH)C=CH), 162.6 ), 133.4 ), 114.4 ), 92.8 (CCl3), 92.0 (Cl3C(NH)C=CH), 24.9\u2005ppm (CH3CO), B\u2010Ar(i) quaternary signal not observed; 11B{1H}\u2005NMR : \u03b4=4.11\u2005ppm; 19F{1H}\u2005NMR : \u03b4=\u2212116.1\u2005ppm; MS(ES\u2212) (m/z): 399.1 \u2212, 400.1 \u2212, 401.1 \u2212, 402.1 \u2212, 403.1 \u2212, 404.1 \u2212; HRMS(ES\u2212) (m/z): [M\u2212H]\u2212 calcd for C17H1211B35Cl319F2NO, 400.0051; found, 400.0048, error: 0.7\u2005ppm.2,2\u2010Bis(4\u2010chlorophenyl)\u20106\u2010methyl\u20104\u2010(trichloromethyl)\u20102,3\u2010dihydro\u20101,3,2\u2010oxazaborinin\u20101\u2010ium\u20102\u2010uide (NBC36): EtOAc/n\u2010hexane, 1:9. Reaction scale: B(OMe)3 , 1\u2010bromo\u20104\u2010chlorobenzene and magnesium turnings gives bis(4\u2010chlorophenyl)borinic acid . Bis(4\u2010chlorophenyl)borinic acid and (Z)\u20104\u2010amino\u20105,5,5\u2010trichloropent\u20103\u2010en\u20102\u2010one gives NBC36 as a yellow solid ; mp: 106\u2013107\u2009\u00b0C; 1H\u2005NMR : \u03b4=7.25 , 7.19 ), 5.76 C=CH), 2.18\u2005ppm ; 13C\u2005NMR : \u03b4=186.7 (CH3CO), 166.3 (Cl3C(NH)C=CH), 133.2 (B\u2010Ar(o)), 133.1 (B\u2010Ar(p)), 127.8 (B\u2010Ar(m)), 92.7 (CCl3), 92.2 (Cl3C(NH)C=CH), 24.8\u2005ppm (CH3CO), B\u2010Ar(i) quaternary signal not observed; 11B{1H}\u2005NMR : \u03b4=3.82\u2005ppm; MS(ES\u2212) (m/z): 431.0 \u2212, 432.0 \u2212, 433.0 \u2212, 434.0 \u2212, 435.0 \u2212, 436.0 \u2212, 437.0 \u2212, 438.0 \u2212; HRMS(ES\u2212) (m/z): [M\u2212H]\u2212 calcd for C17H1211B35Cl5NO, 431.9460; found, 431.9460, error: 0.0\u2005ppm.6\u2010Methyl\u20104\u2010(trichloromethyl)\u20102,2\u2010bis(4\u2010(trifluoromethyl)phenyl)\u20102,3\u2010dihydro\u20101,3,2\u2010oxazaborinin\u20101\u2010ium\u20102\u2010uide (NBC37): EtOAc/n\u2010hexane, 1:19. Reaction scale: B(OMe)3 , 4\u2010bromobenzotrifluoride and magnesium turnings gives bis(4\u2010trifluorophenyl)borinic acid . Bis(4\u2010trifluorophenyl)borinic acid and (Z)\u20104\u2010amino\u20105,5,5\u2010trichloropent\u20103\u2010en\u20102\u2010one gives NBC37 as a yellow solid ; mp: 103\u2013105\u2009\u00b0C; 1H\u2005NMR : \u03b4=7.56 ), 7.46 ), 7.41 , 5.89 C=CH), 2.30\u2005ppm ; 13C\u2005NMR : \u03b4=187.1 (CH3CO), 166.7 (Cl3C(NH)C=CH), 131.8 (B\u2010Ar(o)), 129.2 ), 124.7 , 124.4 ), 92.7 (Cl3C(NH)C=CH), 24.8\u2005ppm (CH3CO), B\u2010Ar(i) and CCl3 quaternary signals not observed; 11B{1H}\u2005NMR : \u03b4=3.56\u2005ppm; 19F{1H}\u2005NMR : \u03b4=\u221262.4\u2005ppm; MS(ES\u2212) (m/z): 499.1 \u2212, 500.1 \u2212, 501.1 \u2212, 502.1 \u2212, 503.1 \u2212, 504.1 \u2212; HRMS(ES\u2212) (m/z): [M\u2212H]\u2212 calcd for C19H1211B35Cl319F6NO, 499.9987; found, 499.9990, error: 0.6\u2005ppm.2,2\u2010Bis(4\u2010methylphenyl)\u20106\u2010methyl\u20104\u2010(trichloromethyl)\u20102,3\u2010dihydro\u20101,3,2\u2010oxazaborinin\u20101\u2010ium\u20102\u2010uide (NBC38): EtOAc/n\u2010hexane, 1:19. Reaction scale: B(OMe)3 , 4\u2010bromotoluene and magnesium turnings gives bis(4\u2010methylphenyl)borinic acid . Bis(4\u2010methylphenyl)borinic acid and (Z)\u20104\u2010amino\u20105,5,5\u2010trichloropent\u20103\u2010en\u20102\u2010one gives NBC38 as a yellow solid ; mp: 105\u2013107\u2009\u00b0C; 1H\u2005NMR : \u03b4=7.41 , 7.28 ), 7.13 ), 5.77 C=CH), 2.34 , 2.23\u2005ppm ; 13C\u2005NMR : \u03b4=186.2 (CH3CO), 136.3 (B\u2010Ar(p)), 131.9 (B\u2010Ar(o)), 128.4 (B\u2010Ar(m)), 91.5 (Cl3C(NH)C=CH), 24.9 (CH3CO), 21.5\u2005ppm (CH3), B\u2010Ar(i) and Cl3C(NH)C=CH quaternary signals not observed; 11B{1H}\u2005NMR : \u03b4=4.61\u2005ppm; MS(ES\u2212) (m/z): 392.1 \u2212, 393.1 \u2212, 394.1 \u2212, 395.1 \u2212, 396.1 \u2212; MS(ES+) (m/z): 301.2 +, 302.1 +, 303.1 +, 304.1 +, 305.1 +, 306.0 +; HRMS(ES\u2212) (m/z): [M\u2212H]\u2212 calcd for C19H1811B35Cl3NO, 392.0553; found, 392.0555, error: 0.6\u2005ppm.2,2\u2010Bis\u20106\u2010methyl\u20104\u2010(trichloromethyl)\u20102,3\u2010dihydro\u20101,3,2\u2010oxazaborinin\u20101\u2010ium\u20102\u2010uide (NBC39): EtOAc/n\u2010hexane, 1:19. Reaction scale: B(OMe)3 , 4\u2010bromo\u20101,2\u2010dichlorobenzene and magnesium turnings gives bisborinic acid . Bisborinic acid and (Z)\u20104\u2010amino\u20105,5,5\u2010trichloropent\u20103\u2010en\u20102\u2010one gives NBC39 as a cream solid ; mp: 109\u2013110\u2009\u00b0C; 1H\u2005NMR : \u03b4=7.37 , 7.36 , 7.10 , 5.88 C=CH), 2.28\u2005ppm , NH signal is observed but overlapping with CHCl3 at 7.26\u2005ppm; 13C\u2005NMR : \u03b4=187.1 (CH3CO), 166.7 (Cl3C(NH)C=CH), 133.5 (B\u2010Ar\u2010C6), 132.1 (B\u2010Ar\u2010C3), 131.1 (B\u2010Ar\u2010C4), 130.9 (B\u2010Ar\u2010C2), 129.9 (B\u2010Ar\u2010C5), 92.8 (Cl3C(NH)C=CH), 92.5 (CCl3), 24.9\u2005ppm (CH3CO), B\u2010Ar\u2010C1 quaternary signal not observed; 11B{1H}\u2005NMR : \u03b4=3.10\u2005ppm; MS(ES\u2212) (m/z): 499.9 \u2212, 500.9 \u2212, 501.9 \u2212, 502.9 \u2212, 503.9 \u2212, 504.9 \u2212, 505.9 \u2212, 506.9 \u2212, 507.9 \u2212; HRMS(ES\u2212) (m/z): [M\u2212H]\u2212 calcd for C17H1011B35Cl7NO, 499.8681; found, 499.8681, error: 0.0\u2005ppm.5\u2010Acetyl\u20106\u2010amino\u20102,2\u2010bis(3\u2010chloro\u20104\u2010methylphenyl)\u20104\u2010(trichloromethyl)\u20102,3\u2010dihydro\u20101,3,2\u2010oxazaborinin\u20101\u2010ium\u20102\u2010uide (NBC40): Using an adapted procedure,iPrMgCl was added dropwise to a solution of 2\u2010chloro\u20104\u2010iodotoluene in anhydrous THF (5\u2005mL) in an oven\u2010dried Schlenk flask under N2. The reaction was stirred at 0\u2009\u00b0C for 5\u2005h. B(OMe)3 was then added and the reaction mixture was stirred overnight allowing to warm to room temperature. HCl was added and the reaction mixture was extracted with EtOAc (3\u00d710\u2005mL), washed with brine (1\u00d710\u2005mL), dried over MgSO4, filtered and evaporated in\u2005vacuo to give crude bis(3\u2010chloro\u20104\u2010methylphenyl)borinic acid as a cream solid in quantitative yield. To a portion of this intermediate in anhydrous THF (5\u2005mL) was added (Z)\u20102\u2010acetyl\u20103\u2010amino\u20104,4,4\u2010trichlorobut\u20102\u2010enamide . The reaction was stirred at 50\u2009\u00b0C for 16\u2005h under N2. The reaction mixture was concentrated and purified by flash column chromatography . Collected fractions were evaporated in\u2005vacuo and stirred in the minimum amount of cold n\u2010hexane for 30\u2005min to induce precipitation. The precipitate was then filtered and dried under vacuum to give NBC40 as a white solid ; mp: 147\u2013148\u2009\u00b0C; 1H\u2005NMR : \u03b4=11.22 C=C), 7.38 , 7.18 , 7.10 , 6.16 , 5.89 , 2.32 , 2.26\u2005ppm ; MS(ES\u2212) (m/z): 502.1 \u2212, 503.1 \u2212, 504.1 \u2212, 505.1 \u2212, 506.1 \u2212, 507.1 \u2212, 508.1 \u2212, 509.1 \u2212; HRMS(ES\u2212) (m/z): [M\u2212H]\u2212 calcd for C20H1711B35Cl5N2O2, 502.9831; found, 502.9833, error: 0.4\u2005ppm.3\u2010Hydroxypyridine\u20102\u2010carbonyloxy\u2010bis(3\u2010chloro\u20104\u2010methylphenyl) borane (AN0128): Prepared according to a previously published method.iPrMgCl was added dropwise to a solution of 2\u2010chloro\u20104\u2010iodotoluene in anhydrous THF (5\u2005mL) in an oven\u2010dried Schlenk flask under N2. The reaction was stirred at 0\u2009\u00b0C for 5\u2005h. B(OMe)3 was then added and the reaction mixture was stirred overnight allowing to warm to room temperature. HCl was then added and the reaction mixture was extracted with EtOAc (3\u00d710\u2005mL), washed with brine (1\u00d710\u2005mL), dried over MgSO4, filtered and evaporated in\u2005vacuo to give crude bis(3\u2010chloro\u20104\u2010methylphenyl)borinic acid as a cream solid in quantitative yield. A portion of this intermediate was dissolved in EtOH (5\u2005mL) and heated at reflux. 3\u2010Hydroxypicolinic acid was added in portions to the hot solution and after the last addition, the reaction mixture was stirred at reflux for 15\u2005min. The reaction was then cooled, resulting in the precipitation of product from the solution. The reaction mixture was partially concentrated and the precipitate was re\u2010crystallised in EtOH to give AN0128 as a white solid ; mp: 166\u2013167\u2009\u00b0C (lit.: 165.0\u2013166.5\u2009\u00b0C)1H\u2005NMR : \u03b4=12.17 , 8.58 , 7.93 , 7.22 , 7.20 , 7.11 , 2.27\u2005ppm ; 13C\u2005NMR : \u03b4=162.2 (COO), 156.2 (Py\u2010C9), 134.2 (Py\u2010C8), 134.0 (Py\u2010C12), 133.0 (B\u2010Ar\u2010C4), 132.1 (B\u2010Ar\u2010C3), 131.9 (B\u2010Ar\u2010C6), 131.5 (Py\u2010C11), 130.7 (B\u2010Ar\u2010C5), 130.5 (B\u2010Ar\u2010C2), 127.5 (Py\u2010C10), 19.4\u2005ppm (CH3), B\u2010Ar\u2010C1 quaternary signal not observed; 11B{1H}\u2005NMR : \u03b4=6.76\u2005ppm; MS(ES\u2212) (m/z): 397.1 \u2212, 398.1 \u2212, 399.1 \u2212, 400.1 \u2212, 401.1 \u2212; HRMS(ES\u2212) (m/z): [M\u2212H]\u2212 calcd for C20H1511B35Cl2NO3, 398.0528; found, 398.0526, error: 0.5\u2005ppm. All data were in agreement with literature values.3\u2010pentane\u20102,4\u2010dione (8): Using a previously reported procedure,3CN was slowly generated by dropwise addition of a solution of trifluoroacetic anhydride in anhydrous pyridine (40\u2005mL) to a solution of trifluoroacetamide in anhydrous pyridine (20\u2005mL) under N2 in a three\u2010neck round\u2010bottom flask equipped with a N2 gas inlet and gas outlet. The gas outlet was connected to a two\u2010neck round\u2010bottom flask containing a solution of acetylacetone and Zn(acac)2 in anhydrous CH2Cl2 (10\u2005mL) that was equipped with a dry ice condenser connected to a bubbler outlet. CF3CN was bubbled into the stirring solution at room temperature for several hours until complete consumption of the trifluoroacetic anhydride solution. The reaction was further stirred at room temperature for 16\u2005h. The reaction mixture was extracted with CH2Cl2 (3\u00d710\u2005mL), washed with brine (1\u00d710\u2005mL), dried over MgSO4, filtered and evaporated in\u2005vacuo to give 8 as a white solid . 1H\u2005NMR : \u03b4=2.46 , 2.18\u2005ppm ; 13C\u2005NMR : \u03b4=202.1 (CH3CO), 196.4 (CH3CO), 145.7 C=C), 120.2 , 112.8 (F3C(NH2)C=C), 32.2 , 29.2\u2005ppm (CH3CO); 19F{1H}\u2005NMR : \u03b4=\u221266.4\u2005ppm. All data were in agreement with literature values.Z)\u20104\u2010Amino\u20105,5,5\u2010trifluoropent\u20103\u2010en\u20102\u2010one (10) was dissolved in EtOH (10\u2005mL) and a saturated solution of K2CO3 (20\u2005mL) was added. The reaction was stirred at 50\u2009\u00b0C for 24\u2005h. The reaction mixture was then extracted with CHCl3 (3\u00d710\u2005mL), washed with brine (1\u00d710\u2005mL), dried over MgSO4, filtered and evaporated in\u2005vacuo. The crude mixture was purified by flash column chromatography to give 10 as an orange solid . 1H\u2005NMR : \u03b4=5.52 C=CH), 2.18\u2005ppm ; 13C\u2005NMR : \u03b4=199.6 (CH3CO), 147.1 C=CH), 120.4 , 94.1 C=CH), 30.5\u2005ppm (CH3CO); 19F{1H}\u2005NMR : \u03b4=\u221271.8\u2005ppm. All data were in agreement with literature values.6\u2010Methyl\u20102,2\u2010diphenyl\u20104\u2010(trifluoromethyl)\u20102,3\u2010dihydro\u20101,3,2\u2010oxazaborinin\u20101\u2010ium\u20102\u2010uide (NBC41): Using an adapted procedure,10 was added to a solution of DPBA in anhydrous THF (5\u2005mL). The reaction mixture was stirred at 50\u2009\u00b0C under Ar for 16\u2005h. The reaction mixture was then concentrated and purified by flash column chromatography to give NBC41 as a yellow solid ; mp: 96\u201397\u2009\u00b0C (lit.: 99\u2013100\u2009\u00b0C);1H\u2005NMR : \u03b4=7.14\u20137.31 , 6.95 , 5.44 C=CH), 2.17\u2005ppm ; 13C\u2005NMR : \u03b4=188.3 (CH3CO), 156.7 C=CH), 131.8 (B\u2010Ph(o)), 127.6 (B\u2010Ph(m)), 127.1 (B\u2010Ph(p)), 118.8 , 91.6 (F3C(NH)C=CH), 24.9\u2005ppm (CH3CO); 11B{1H}\u2005NMR : \u03b4=4.37\u2005ppm; 19F{1H}\u2005NMR : \u03b4=\u221272.9\u2005ppm; MS(ES\u2212) (m/z): 316.11 \u2212; MS(ES+) (m/z): 340.11 +; HRMS(ES\u2212) (m/z): [M+Na]\u2212 calcd for C17H1511BF3NO, 340.1091; found, 340.1085, error: 1.8\u2005ppm. All data were in agreement with literature values.Z)\u20102\u2010Acetyl\u20103\u2010amino\u20104,4,4\u2010trifluorobut\u20102\u2010enamide (9) in anhydrous pyridine (40\u2005mL) to a solution of trifluoroacetamide in anhydrous pyridine (20\u2005mL) under N2 in a three\u2010neck round\u2010bottom flask equipped with a N2 gas inlet and gas outlet. The gas outlet was connected to a two\u2010neck round\u2010bottom flask containing a solution of acetoacetamide and Zn(acac)2 in anhydrous CH2Cl2 (10\u2005mL) that was equipped with a dry ice condenser connected to a bubbler outlet. CF3CN was bubbled into the stirring solution at room temperature for several hours until complete consumption of the trifluoroacetic anhydride solution. The reaction was further stirred at room temperature for 16\u2005h. The reaction mixture was extracted with EtOAc (5\u00d750\u2005mL), washed with brine (3\u00d750\u2005mL), dried over MgSO4, filtered and evaporated in\u2005vacuo. Et2O (10\u2005mL) was added to induce precipitation and the precipitate was filtered, washed with additional Et2O and dried to give 9 as a white solid ; mp: 192\u2013193\u2009\u00b0C; 1H\u2005NMR : \u03b4=9.06 , 7.75 , 7.40 , 2.18\u2005ppm ; 13C\u2005NMR : \u03b4=196.5 (CH3CO), 168.3 (CONH2), 144.9 C=C), 120.3 , 107.0 (F3C(NH2)C=C), 28.0\u2005ppm (CH3CO); 19F{1H}\u2005NMR : \u03b4=\u221265.4\u2005ppm; MS(ES\u2212) (m/z): 195.04 \u2212; MS(ES+) (m/z): 219.03 +; HRMS(ES\u2212) (m/z): [M+Na]\u2212 calcd for C6H7F3N2O2, 219.0352; found, 219.0346, error: 2.7\u2005ppm.5\u2010Acetyl\u20106\u2010amino\u20102,2\u2010diphenyl\u20104\u2010(trifluoromethyl)\u20102,3\u2010dihydro\u20101,3,2\u2010oxazaborinin\u20101\u2010ium\u20102\u2010uide (NBC42): 9 was added to a solution of DPBA in anhydrous THF (5\u2005mL). The reaction mixture was stirred at 50\u2009\u00b0C under Ar for 16\u2005h. The reaction mixture was then concentrated and purified by flash column chromatography . The collected fractions were combined, evaporated in\u2005vacuo and precipitated in n\u2010hexane (10\u2005mL) to give NBC42 as a white solid ; mp: 118\u2013120\u2009\u00b0C; 1H\u2005NMR : \u03b4=10.05 , 7.22\u20137.36 , 6.17 , 2.26\u2005ppm ; 13C\u2005NMR : \u03b4=196.3 (CH3CO), 169.8 (CONH2), 156.7 C=C), 131.8 (B\u2010Ar(o)), 127.5 (B\u2010Ar(m)), 127.0 (B\u2010Ar(p)), 119.4 , 97.2 (F3C(NH)C=C), 30.5\u2005ppm , B\u2010Ar(i) quaternary signal not observed; 11B{1H}\u2005NMR : \u03b4=2.55\u2005ppm; 19F{1H}\u2005NMR : \u03b4=\u221265.1\u2005ppm; MS(ES\u2212) (m/z): 227.0 \u2212, 359.1 \u2212, 523.3 \u2212; MS(ES+) (m/z): 219.0 +, 399.1 +, 219.0 +, 563.2 +; HRMS(ES+) (m/z): [M+H]+ calcd for C18H1711BF3N2O2, 361.1330; found, 361.1338, error: 2.3\u2005ppm.Cell culture: Immortalised murine bone marrow\u2010derived macrophages (iBMDMs) were cultured in DMEM, 10\u2009% fetal bovine serum (FBS), 100\u2005U\u2009mL\u22121 penicillin and 100\u2005\u03bcg\u2009mL\u22121 streptomycin (PenStrep). Cells were seeded overnight at 0.75\u00d7106 cells per mL and then stimulated with LPS , and then incubated with vehicle (0.5\u2009% DMSO) or drug as indicated (10\u2005\u03bcm) for 15\u2005min before activation of NLRP3 using nigericin . IL\u20101\u03b2 release was measured by a specific ELISA (R&D systems).Data presentation and statistical analysis: Data are presented as mean values \u00b1 standard error of the mean (SEM) of at least three separate experiments. Statistical analyses performed were one\u2010way analysis of variance (ANOVA) with Dunnett's multiple comparisons test post hoc. Accepted levels of significance were *p<0.05, ***p<0.001. Statistical analyses were carried out using GraphPad Prism.The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "Hydrogen bonding through the water mol\u00adecules gives two-dimensional sheets lying parallel to (100).The present paper describes the synthesis and crystal structure of a cobalt(II) isobutyrate dihydrate, based on a slightly distorted CoO 3)2CO2}2(H2O)]\u00b7H2O}n, the Co2+ ion is hexa\u00adcoordinated in a slightly distorted octa\u00adhedral coordination environment defined by two O atoms from two bridging water mol\u00adecules and four O atoms from four bridging carboxyl\u00adate ligands. The carboxyl\u00adates adopt two different coordination modes, \u03bc-(\u03ba2O:O\u2032) and \u03bc-(\u03ba2O:O), forming a one-dimensional polymeric chain extending along [010]. The intra-chain cobalt\u22efcobalt separation is 3.2029\u2005(2)\u2005\u00c5. The polymeric chains are linked by hydrogen bonds involving the water mol\u00adecules of solvation, giving a two-dimensional network structure lying parallel to (100).In the title cobalt(II) coordination polymer with isobutyrate ligands, {[Co{CH(CH Oligo- and polynuclear cobalt carboxyl\u00adates in turn have attracted great attention because of their utilization in homogeneous oxidation catalysis 2CO2}2(H2O)]\u00b7H2O}n, (I)As a part of our ongoing studies on these compounds, we describe here synthesis and crystal structure of the title compound, {[Co{CH(CH2+ cation coordinated by four O atoms from four bridging isobutyrate ligands and two O atoms from two bridging water mol\u00adecules (O1W) in a distorted octa\u00adhedral coordination. A water mol\u00adecule of solvation (O2W) is also present \u2013110.31\u2005(2)\u00b0. This data correlates with the angles and the distances in cobalt(II) acetate dihydrate which has a similar structure nt Fig.\u00a01. The Co\u2014nt Fig.\u00a01 and the \u221e[Co(H2O)((CH3)2CHCOO)2] composition, extending along [010] and \u03bc-(\u03ba2O:O). The C\u2014O bond lengths of the first group (involving O1A and O2A) have close values [1.2755\u2005(10) and 1.2533\u2005(10)\u2005\u00c5], whereas those of the second group (involving O1B and O2B) have a more striking difference [1.2878\u2005(9) and 1.2510\u2005(11)\u2005\u00c5]. The carboxyl\u00adate O2B atom of the second group forms an inter-unit hydrogen bond with the bridging water mol\u00adecule [O1W\u2014H\u22efO2Bi = 2.6206\u2005(9)\u2005\u00c5] 0] Fig.\u00a02. The Co\u22ef\u00c5] Fig.\u00a02.W) by a system of hydrogen bonds, forming a sheet structure arranged parallel to (100) Table\u00a02. Only weet al., 2016\u221e[Co(RCOO)2(H2O)]: acetate was added. The reaction mixture was period\u00adically stirred in an ultrasonic bath at room temperature until the liberation of carbon dioxide ceased. The unreacted CoCO3\u00b76H2O was removed by filtration, and the filtrate was allowed to stand at room temperature for slow evaporation. Red single crystals of (I)The title compound was synthesized using a similar procedure as for the synthesis of the analogous carboxyl\u00adates cobalt(II) propionate dihydrate (Fischer Uiso(H) set to 1.5Ueq(O). Other hydrogen atoms were placed in calculated positions and refined using a riding model with d(C\u2014H) = 0.98\u2005\u00c5, Uiso(H) = 1.2Ueq(C) for the tertiary carbon atoms and d(C\u2014H) = 0.96\u2005\u00c5, Uiso(H) = 1.5Ueq(C) for the methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017001360/zs2371sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 1529830CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "C=O\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions also occur. Hirshfeld surface analysis was used to confirm and qu\u00adantify the supra\u00admolecular inter\u00adactions.The structure of a coumarin ester is reported and compared with the results of a quantum chemical calculation. In the crystal, inter\u00admolecular C\u2014H\u22efO contacts generate an infinite C(6) chain along the 20H18O4, the benzoate ring is oriented at an acute angle of 33.10\u2005(12)\u00b0 with respect to the planar (r.m.s deviation = 0.016\u2005\u00c5) coumarin ring system. An intra\u00admolecular C\u2014H\u22efO hydrogen bond closes an S(6) ring motif. In the crystal, C\u2014H\u22efO contacts generate infinite C(6) chains along the b-axis direction. Also present are \u03c0\u2013\u03c0 stacking inter\u00adactions between neighbouring pyrone and benzene rings [centroid\u2013centroid distance = 3.7034\u2005(18)\u2005\u00c5] and C=O\u22ef\u03c0 inter\u00adactions [O\u22efcentroid = 3.760\u2005(3)\u2005\u00c5]. The data obtained from quantum chemical calculations performed on the title compound are in good agreement with the observed structure, although the calculated C\u2014O\u2014C\u2014C torsion angle between the coumarin ring system and the benzoate ring (129.1\u00b0) is somewhat lower than the observed value [141.3\u2005(3)\u00b0]. Hirshfeld surface analysis has been used to confirm and qu\u00adantify the supra\u00admolecular inter\u00adactions.In the title compound, C Coumarin and its derivatives have been reported to serve as anti-bacterial ring motif arises from an intra\u00admolecular C6\u2014H6\u22efO4 hydrogen bond, and generates a pseudo-tricyclic ring system em Fig.\u00a01. The couc-axis direction, as shown in Fig.\u00a03B = 2.38\u2005\u00c5] is found at a distance shorter than the sum of the van der Waals radii. An unusual C10=O4\u22ef\u03c0 inter\u00adaction , is also present. The resulting supra\u00admolecular aggregation is completed by the presence of \u03c0\u2013\u03c0 stacking = 3.7035\u2005(18) and Cg3\u22efCg1 = 3.7034\u2005(18)\u2005\u00c5, where Cg1 and Cg3 are the centroids of the pyrone and the C11\u2013C16 benzene rings, respectively] that are less than 3.8\u2005\u00c5, the maximum regarded as suitable for an effective \u03c0\u2013\u03c0 inter\u00adaction and 3.6143\u2005(13)\u2005\u00c5, respectively, and the distances between Cg1 and a perpendicular projection of Cg3 on ring 1 (slippage) are 0.726 and 0.807\u00c5, respectively.In the crystal, two types of inter\u00admolecular hydrogen-bonding inter\u00adactions are present Table\u00a01. The C8\u2014ng Fig.\u00a04 between et al., 2016et al.,1982et al., 1985et al., 2011et al., 2014meta-substituted coumarin esters dnorm surfaces mapped over a fixed colour scale of \u22120.39 (red) to 1.4\u2005\u00c5 (blue) with the program CrystalExplorer 3.1 dnorm highlights six red spots showing distances shorter than the sum of the van der Waals radii. These dominant inter\u00adactions correspond to inter\u00admolecular C\u2014H\u22efO hydrogen bonds, O\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions between the surface and the neighbouring environment. The mapping also shows white spots with distances equal to the sum of the van der Waals radii and blue regions with distances longer than the sum of the van der Waals radii. The surfaces are transparent to allow visualization of the mol\u00adecule , as shown by the X-ray study. The most significant contribution to the Hirshfeld surface (46.8%) is from H\u22efH contacts, which appear in the central region of the FP . H\u22efO/O\u22efH inter\u00adactions with a 24.1% contribution appear as blue spikes in Fig.\u00a06c and show the presence of O\u22efH contacts, whereas the C\u22efH/H\u22efC plot (17.3%) gives information about inter\u00admolecular hydrogen bonds . Other visible spots in the Hirshfeld surfaces show C\u22efO/O\u22efC and O\u22efO contacts, which contribute only 4.0 and 1.0%, respectively .Mol\u00adecular Hirshfeld surfaces of 2-oxo-2le Fig.\u00a05. Furtherns Fig.\u00a06a, as shFP Fig.\u00a06b. H\u22efO/Ods Fig.\u00a06d. Otherly Fig.\u00a06e and 6f++G basis set. The crystal structure in the solid state was used as the starting structure for the calculations. The DFT calculations are performed with the GAUSSIAN09 program package tert-butyl\u00adbenzoyl chloride in dry tetra\u00adhydro\u00adfuran (30 to 40\u2005ml), was added dry tri\u00admethyl\u00adamine and 7-hy\u00addroxy\u00adcoumarin in small portions over 30\u2005min. The mixture was then refluxed for four\u2005h and poured into 40\u2005ml of chloro\u00adform. The solution was acidified with diluted hydro\u00adchloric acid until the pH was 2\u20133. The organic layer was extracted, washed with water to neutrality, dried over MgSO4 and the solvent removed. The resulting precipitate (crude product) was filtered off with suction, washed with petroleum ether and recrystallized from chloro\u00adform. Colourless crystals of the title compound were obtained in a good yield: 90%; m.p. 406\u2013408\u2005K.To a solution of 4-Uiso(H) constrained to 1.2 (aromatic) or 1.5 (meth\u00adyl) times Ueq(C) of the respective parent atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018004188/sj5549sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018004188/sj5549Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018004188/sj5549Isup3.cmlSupporting information file. DOI: 1828991CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "ASBTapical sodium\u2010dependent bile acid transporterBAbile acidBSEPbile salt export pumpCAcancer antigenCDCAchenodeoxycholic acidCholcholesterolCol1\u03b1, collagen 1\u03b1; COX2cyclooxygenase 2eNOSendothelial NO synthaseET1endothelin 1FAfatty acidFFAfree fatty acidFGFfibroblast growth factorFGFRFGF receptorFXRfarnesoid X receptorHCChepatocellular carcinomaHDLhigh\u2010density lipoproteinIFNinterferonIGF1Rinsulin\u2010like growth factor 1 receptorILinterleukiniNOSinducible NO synthaseKCKupffer cellLDLlow\u2010density lipoproteinLDLrLDL receptorLp(a), lipoprotein a; LSECliver sinusoidal endothelial cellMCP1monocyte chemoattractant protein 1MDRmultidrug resistanceMMPmatrix metalloproteinaseNAFLDnonalcoholic fatty liver diseaseNASHnonalcoholic steatohepatitisNF\u03baBnuclear factor kappa BNOnitric oxideNTCPsodium/taurocholate cotransporting polypeptidePLphospholipidsSR\u2010BIscavenger receptor class B type 1p\u2010STAT3phosphorylated signal transducer and activator of transcription 3TGtriglycerideTGFtransforming growth factorTIMPtissue inhibitor of metalloproteinaseTJtight junctionTNFtumor necrosis factorVCAMvascular cell adhesion moleculeVLDLvery low\u2010density lipoproteinZO\u20101zona occludens 1."} +{"text": "Offset \u03c0\u2013\u03c0 inter\u00adactions are also present.The title hydrated salt, C 10H12N3O3S+\u00b7Cl\u2212\u00b7H2O, the benzimidazolium ring system is almost planar (r.m.s. deviation = 0.006\u2005\u00c5) and the nitro group is inclined at an angle of 4.86\u2005(9)\u00b0 to this plane. In the crystal, C\u2014H\u22efO hydrogen bonds form centrosymmetric R22(20) dimers and these are further aggregated through N\u2014H\u22efO and O\u2014H\u22efCl hydrogen bonds involving the water mol\u00adecules and chloride anions. Aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions are also found between two parallel benzene rings or the benzene and imidazolium rings, with centroid\u2013centroid distances of 3.5246\u2005(9) and 3.7756\u2005(9)\u2005\u00c5, respectively. Analysis of the bond lengths and comparison with related compounds show that the nitro substituent is not involved in conjugation with the adjacent \u03c0-system and hence has no effect on the charge distribution of the heterocyclic ring.In the cation of the title hydrated molecular salt, C The centroid\u2013centroid separations are less than 3.8\u2005\u00c5, the maximum regarded as suitable for an effective \u03c0\u2013\u03c0 inter\u00adaction in dimethyl sulfoxide (DMSO) (10\u2005ml). The reaction mixture was agitated for 5\u2005h at room temperature. 50\u2005ml of water was then added to the reaction mixture, and the products were extracted with di\u00adchloro\u00admethane (3 \u00d7 50\u2005ml). The combined organic extracts were washed with ammonium chloride solution (10\u2005g of ammonium chloride in 100\u2005ml of water), dried over Na2SO4, filtered and evaporated under reduced pressure. The residue was purified by column chromatography on silica gel . The resulting powder was dissolved in di\u00adchloro\u00admethane and after three days, yellow crystals suitable for single-crystal X-ray diffraction analysis were obtained in 72% yield with a melting point of 425\u2005K.2-Chloro\u00adethanol and potassium carbonate were added to 2-methyl\u00adthio-5-nitro-11H NMR \u03b4(p.p.m.): 2.7 ; 3 ; 3.7 ; 4.3 ; 5 ; 7.5\u20138.5 .13C \u03b4 (p.p.m.): 114.28 (CH3); 47 (CH2O); 59 (CH2N); 106.56; 110.03; 112.87; 117.13; 136.38; 147.37; 155.52 ; 162.23 (C=N).Uiso(H) were refined with O\u2014H distances restrained to be 0.82\u2005\u00c5 with a standard deviation of 0.02\u2005\u00c5. Other H atoms were placed in calculated positions and refined using a riding-model approximation with Uiso(H) constrained to 1.2 or 1.5 times Ueq of the respective parent atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016013657/sj5502sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989016013657/sj5502Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016013657/sj5502Isup3.cmlSupporting information file. DOI: 1500918CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Mol\u00adecules (I) and (II) are nearly planar, while mol\u00adecule (III) is not planar. In compounds (I) and (II), mol\u00adecules are linked into chain by C\u2014H\u22ef\u03c0 inter\u00adactions. In compound (III), mol\u00adecules are linked by a pair of C\u2014H\u22efO hydrogen bonds, forming inversion dimers. Weak C\u2014Br\u22ef\u03c0 inter\u00adactions are present in (III). viz. (E)-1-[4-(4-bromo\u00adbut\u00adoxy)\u00adphen\u00adyl]-3-phenyl\u00adprop-2-en-1-one, C19H19BrO2, (I), (E)-1-[4-(4-bromo\u00adbut\u00adoxy)\u00adphen\u00adyl]-3-(4-meth\u00adoxy\u00adphen\u00adyl)prop-2-en-1-one, C20H21BrO3, (II), and (E)-1-[4-(4-bromo\u00adbut\u00adoxy)\u00adphen\u00adyl]-3-prop-2-en-1-one, C21H23BrO4, (III), are reported. In all mol\u00adecules, the conformation of the keto group with respect to the olefinic bond is s-cis. Mol\u00adecules of (I) and (II) are nearly planar, while mol\u00adecule (III) is not planar. In the crystal of compounds (I) and (II), mol\u00adecules are linked into chains parallel to the c axis by C\u2014H\u22ef\u03c0 inter\u00adactions. In the crystal of compound (III), mol\u00adecules are linked by a pairs of C\u2014H\u22efO hydrogen bonds, forming inversion dimers. Weak C\u2014Br\u22ef\u03c0 inter\u00adactions are also observed in (III).The crystal structures of three chalcones with a bromo-substituted but\u00adoxy side chain, D\u2014\u03c0\u22efA type mol\u00adecule. When the electron-rich groups are located on the 4 and/or 4\u2032 positions, the electron flow follows a \u039b-shaped path, and therefore the mol\u00adecule is called a \u039b-shaped mol\u00adecule anti-tumor activity connected by an enone moiety with a bromo\u00adbut\u00adoxy side chain attached at the 4-position of one of the phenyl rings. In mol\u00adecule (I)B. All of them crystallize in the monoclinic space group P21/c with Z = 4. All three mol\u00adecules adopt an s-cis conformation about the central olefinic C12=C13 bond with O2\u2014C11\u2014C12\u2014C13 torsion angles of \u22123.2\u2005(4), \u22121.6\u2005(5) and \u221221.5\u2005(4)\u00b0, respectively, and the hydrogen atoms of the central enone groups are trans-arranged with respect to the C12=C13 double bond. Mol\u00adecules (I)A and B are 3.10\u2005(13), 5.34\u2005(11)\u00b0 in compound (I)et al., 2007B with torsion angles of \u22123.3\u2005(4)\u00b0 (C20\u2014O3\u2014C17\u2014C16) in mol\u00adecule (II)The mol\u00adecular structures of the title compounds (I)A with a C6\u2014C5\u2014O1\u2014C4 torsion angle of 179.7\u2005(3)\u00b0.In compounds (I)et al., 2003In compounds (I)et al., 1995et al., 2013et al., 2005via atom H2A of ring A and the centroid of ring B of a symmetry-related mol\u00adecule (Table\u00a01c axis. In compound (II)c axis by two C\u2014H\u22ef\u03c0 inter\u00adactions involving the C2 and C3 carbon atoms via atoms H2B and H3A of ring A and the centroid of ring B of two symmetry-related mol\u00adecules (Table\u00a02The packing for mol\u00adecules (I)e Table\u00a01, formings Table\u00a02.A via atom H10 and the O3 oxygen atom \u2005\u00c5; Cg is the centroid of ring B; symmetry code: (ii) 1\u00a0\u2212\u00a0x, \u2212y, z] having a \u2018face-on\u2019 geometry m Table\u00a03. In addiet al., 2016A search of the Cambridge Structural Database with benzaldehyde (1 equiv.) for compound (I)2Cl2 (3 \u00d7 100\u2005ml). The organic layers were separated, washed with brine (1 \u00d7 150\u2005ml), dried over anhydrous Na2SO4 and evaporated to give the crude bromo compounds, which were purified by column chromatography (SiO2) using a mixture of hexa\u00adne/CHCl3 (9:2 v/v) as eluent to afford yellow solids. The compounds were recrystallized by slow evaporation of chloro\u00adform solutions.Mixtures of chalcone (1 equiv.), 1,4-di\u00adbromo\u00adbutane (1.2 equiv.) and anhydrous potassium carbonate (2 equiv.) in dry acetone (40\u2005mL) were then stirred at 333\u2005K for 12\u2005h. After completion of reactions, the solvents were evaporated under reduced pressure and the residues extracted with CHUiso(H) values were set to 1.2Ueq(C) or 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989017010052/rz5217sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989017010052/rz5217Isup5.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017010052/rz5217IIsup6.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989017010052/rz5217IIIsup7.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989017010052/rz5217Isup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989017010052/rz5217IIsup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989017010052/rz5217IIIsup7.cmlSupporting information file. DOI: 1545201, 1545200, 1545196CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Proc. R. Soc. A472, 20160062 (Published online May 4 2016) (doi:10.1098/rspa.2016.0062)K is symmetrical with respect to \u03b7. More generally they should be replaced by periodic boundary conditions on \u2014\u2003The Neumann boundary conditions (3.6), (3.9), (3.26), (A\u200911), (A\u200915), (A\u200922), (A\u200927) and (A\u200931) are applicable only when \u2014\u2003Equation (4.25) should read\u2014\u2003Equations (4.26) and (4.27) should read\u2014\u2003The two lines following equation (4.30) should read:\u03a9+ with \u0393, and \u03a9+ with \u0393, with both where \u2014\u2003The line following equation (4.33) should read:\u03a9\u2212 with \u0393, andwhere \u2014\u2003Equation (A\u200918) should read:"} +{"text": "III ions (each with site symmetry 2..) in the title compound have a distorted octa\u00adhedral coordination environment with four N atoms of a cyclam ligands and two chloride ions or one oxalate bidentate ligand in a cis position whereby the cyclam ligands adopt a cis-V conformation. The crystal packing is stabilized by extensive hydrogen-bonding inter\u00adactions among the cyclam N\u2013H groups, the Cl ligands, and O atoms of the oxalate and ClO4\u2212 anions.Two Cr 2(C10H24N4)][Cr(C2O4)(C10H24N4)](ClO4)2 , there are two independent halves of the [CrCl2(cyclam)]+ and [Cr(ox)(cyclam)]+ cations, and one perchlorate anion. In the complex cations, which are completed by application of twofold rotation symmetry, the CrIII ions are coordinated by the four N atoms of a cyclam ligand, and by two chloride ions or one oxalate bidentate ligand in a cis arrangement, displaying an overall distorted octa\u00adhedral coordination environment. The Cr\u2014N(cyclam) bond lengths are in the range of 2.075\u2005(5) to 2.096\u2005(4)\u2005\u00c5 while the Cr\u2014Cl and Cr\u2014O(ox) bond lengths are 2.3358\u2005(14) and 1.956\u2005(4)\u2005\u00c5, respectively. Both cyclam moieties adopt the cis-V conformation. The slightly distorted tetra\u00adhedral ClO4\u2212 anion remains outside the coordination sphere. The supra\u00admolecular architecture includes N\u2014H\u22efO and N\u2014H\u22efCl hydrogen bonding between cyclam NH donor groups, O atoms of the oxalate ligand or ClO4\u2212 anions and one Cl ligand as acceptors, leading to a three-dimensional network structure.In the asymmetric unit of the title compound, [CrCl Each cyclam moiety exhibits point group symmetry ..2 and can be described as being in the cis-V (anti\u2013anti) conformation 2(cyclam)]ClO4 [2.069\u2005(3)\u20132.103\u2005(3)\u2005\u00c5] 2(cyclam)]NO2 [2.0874\u2005(16)\u20132.0916\u2005(15)\u2005\u00c5] 2\u00b70.5H2O [2.070\u2005(5)\u20132.089\u2005(5)\u2005\u00c5] 2(cyclam)]NCS [2.0851\u2005(14)\u20132.0897\u2005(14)\u2005\u00c5] 2(cyclam)]ClO4 [2.046\u2005(2)\u20132.060\u2005(2)\u2005\u00c5] 2(cyclam)]BF4 [2.064\u2005(4)\u20132.073\u2005(4)\u2005\u00c5] 2(cyclam)][ZnCl4]Cl\u00b7H2O [2.0501\u2005(15)\u20132.0615\u2005(15)\u2005\u00c5] 2(cyclam)]ClO4 [2.058\u2005(4)\u20132.064\u2005(4)\u2005\u00c5] 2]2ZnCl4 2(Me2tn)2]ClO4\u00b72H2O 2(Me2tn)2]SCN\u00b70.5H2O F2]ClO4 \u2005\u00c5 for the oxalate ligand is close to the mean of 1.959\u2005(4)\u2005\u00c5 found in [Cr(ox)(cyclam)]ClO4 \u2005\u00c5 is comparable to those found in cis-[CrCl2(cyclam)]ClO4 [2.331\u2005(2)\u2005\u00c5] ]ClO4 [2.3157\u2005(7)\u2005\u00c5] 2]2ZnCl4 [2.3112\u2005(6)\u2005\u00c5] 2]Cl [2.3253\u2005(7)\u2005\u00c5] \u00b0, while the Cl1B\u2014Cr1B\u2014ClBi angle is 89.11\u2005(9)\u00b0 [symmetry code: (i) \u2013x\u00a0+\u00a0y\u00a0+\u00a0z]. The folded angles of the cyclam in [CrCl2(cyclam)]+ and [Cr(ox)(cyclam)]+ cations are 93.7\u2005(2) and 97.5\u2005(2)\u00b0, respectively. The significant distortion of the octa\u00adhedral coordination sphere and the larger folded angle in the [Cr(ox)(cyclam)] + cation seem to arise from the small bite angle of the oxalato ligand. The tetra\u00adhedral ClO4\u2212 anion remains outside the coordination sphere of two CrIII ions. It is distorted due to its involvement in hydrogen-bonding inter\u00adactions. Cl\u2014O bond lengths range from 1.426\u2005(5) to 1.443\u2005(5)\u2005\u00c5 and the O\u2014Cl\u2014O angles from 107.8\u2005(4)\u2013111.0\u2005(3)\u00b0.The asymmetric unit contains two halves of the [CrClon Fig.\u00a01. In eachon Fig.\u00a01. The Cr\u2014+ cation while N\u2014H\u22efO and N\u2014H\u22efCl contacts inter\u00adconnect two [Cr(ox)(cyclam)]+ and one cis-[CrCl2(cyclam)]+ cation (cyclam)]n Table\u00a01 and 3 \u25b8.et al., 2016cis-[CrL2(C10H24N4)]+ unit. The crystal structure of cis-[CrCl2(cyclam)]ClO4 2(cyclam)]ClO4 2(cyclam)](ClO4)Cl2 2)(cyclam)]NO2 2\u00b70.5H2O 2(cyclam)]NCS ][Cr(ox)(cyclam)]2+ with any anion has been deposited.A search of the Cambridge Structural Database ]ClO4 and [Cr(ox)(cyclam)]ClO4, were prepared according to literature methods ][Cr(ox)(cyclam)](ClO4)2, was prepared by mixing concentrated equimolar aqueous solutions of the two starting compounds. A saturated solution of NaClO4 was added to the resulting solution for crystallization, and allowed to stand at room temperature for two days to give needle-like orange crystals of (I)The free ligand cyclam was purchased from Fluka and used as provided. All chemicals were reagent grade materials and were used without further purification. The starting materials, Uiso(H) values of 1.2Ueq of the parent atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016014134/wm5323sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016014134/wm5323Isup2.hklStructure factors: contains datablock(s) I. DOI: 1502530CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Secondary Sn\u22efS inter\u00adactions lead to centrosymmetric dimeric aggregates in the crystal.In (CH 3)2(C5H8NOS2)2], has the SnIV atom bound by two methyl groups which lie over the weaker Sn\u2014S bonds formed by two asymmetrically chelating di\u00adthio\u00adcarbamate ligands so that the coordination geometry is skew-trapezoidal bipyramidal. The most prominent feature of the mol\u00adecular packing are secondary Sn\u22efS inter\u00adactions [Sn\u22efS = 3.5654\u2005(7)\u2005\u00c5] that lead to centrosymmetric dimers. These are connected into a three-dimensional architecture via methyl\u00adene-C\u2014H\u22efS and methyl-C\u2014H\u22efO(morpholino) inter\u00adactions. The Sn\u22efS inter\u00adactions are clearly evident in the Hirshfeld surface analysis of the title compound along with a number of other inter\u00admolecular contacts.The title compound, [Sn(CH Fewer examples are known for five-coordinate, trigonal\u2013bipyramidal species, e.g. (t-Bu)2Sn(S2CNMe2)2 in which one di\u00adthio\u00adcarbamate ligand is monodentate CH2CH2]2Sn(S2CNMe)2 where the carbonyl-O atom of one Sn-bound organic substituent is also coordinating the tin atom 2 structures as the strong chelating ability of the di\u00adthio\u00adcarbamate ligand reduces the Lewis acidity of the tin atom. However, in (I)Both binary tin and organotin di\u00adthio\u00adcarbamates, IV atom in the title compound (I)R2Sn(S2CNRR\u2019)2 mol\u00adecules, i.e. skew-trapezoidal bipyramidal rather than octa\u00adhedral \u2212 d\u00b0 at the tin atom. The angle subtended at the tin atom by the strongly bound sulfur atoms of 85.878\u2005(19)\u00b0 is significantly less than that formed by the weakly bound sulfur atoms, i.e. 143.066\u2005(18)\u00b0, and is largely responsible for the formation of the skew-trapezoidal plane about the tin atom.The Sna, where two long edges of the translationally displaced trapezoidal planes approach each other to form the inter\u00adactions. Here, Sn\u22efS4i is 3.5654\u2005(7)\u2005\u00c5, which is approximately 0.4\u2005\u00c5 shorter than the sum of the van der Waals radii of Sn and S of 3.97\u2005\u00c5 , Table\u00a02b.An inter\u00adesting feature of the mol\u00adecular packing in (I)et al., 2017dnorm in Fig.\u00a03a indicate the formation of the supra\u00admolecular dimer through secondary Sn\u22efS contacts. On the Hirshfeld surface mapped over electrostatic potential in Fig.\u00a04a. The pair of bright-red spots appearing near the methyl-H12C and morpholine-O1 atoms in Fig.\u00a03b represent the respective donor and acceptor atoms of the C12\u2014H\u22efO1 inter\u00adaction. The comparatively weaker methyl\u00adene-C10\u2014H\u22efS1 inter\u00adaction is viewed as a pair of faint-red spots near these atoms in Fig.\u00a03b. It is important to note from the immediate environments about a reference mol\u00adecule within dnorm-mapped Hirshfeld surfaces highlighting inter\u00admolecular inter\u00adactions in Fig.\u00a05The Hirshfeld surfaces calculated on the structure of (I)a, and those delineated into H\u22efH, S\u22efH/H\u22efS, O\u22efH/H\u22efO, C\u22efH/H\u22efC, N\u22efH/H\u22efN, Sn\u22efS/S\u22efSn and S\u22efS contacts 2. Of these, 12 feature secondary Sn\u22efS inter\u00adactions which, with (I)R2Sn(S2CNRR\u2019)2 structures have Sn\u22efS secondary inter\u00adactions. Selected geometric details for the 13 structures are collated in Table\u00a05A, is a dimeric aggregate disposed about a centre of inversion, as is in (I)i.e. nine. This motif is illustrated in Fig.\u00a07a for (PhCH2)2Sn(S2CNEt2)2 CH2C6H4N-4)2 ligands and self-assemble into supra\u00admolecular chains. In {Me2SnS2CN(CH2Ph)CH2CH2(PhCH2)NCS2SnMe2}2 CH2CH2(PhCH2)NCS2SnMe2}2, where there are two independent, centrosymmetric mol\u00adecules in the asymmetric unit. Here, the resulting supra\u00admolecular chain is twisted \u22121): 1465(s), 1423(s) \u03bd(C\u2014N), 1222(s) \u03bd(C\u2014O), 1110(m), 994(s) \u03bd(C\u2014S), 541(m) \u03bd(Sn\u2014C) cm\u22121. 1H NMR (CDCl3): 4.18 , 3.77 , 1.54 .Sodium morpholine\u00addithio\u00adcarbamate (prepared from the reaction between carbon di\u00adsulfide and morpholine (Merck) in the presence of sodium hydroxide; 1.0\u2005mmol, 0.185\u2005g) in methanol (20\u2005ml) was added to di\u00admethyl\u00adtin dichloride in methanol (10\u2005ml). The resulting mixture was stirred and refluxed for 2\u2005h. The filtrate was evaporated until an off-white precipitate was obtained. The precipitate was recrystallized from methanol solution by slow evaporation to yield colourless prisms. Yield: 0.305\u2005g, 64.4%; m.p.: 448\u2005K. IR (cmUiso(H) set to 1.2\u20131.5Ueq(C). Owing to poor agreement, one reflection, i.e. I, global. DOI: 10.1107/S2056989017006855/hb7675Isup2.hklStructure factors: contains datablock(s) I. DOI: 1548414CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Diabetic neuropathy is the most common complication of diabetes. The idea of alterations in energy metabolism in diabetes is emerging. The biogenic antioxidant R(+)-thioctic acid has been successfully used in the treatment of diabetic polyneuropathic (DPN) patients. The effects of R(+)-thioctic acid administration were evaluated in 12 DPN patients at baseline and at 15, 30, 60, and 120 administration days throughout the assessment of oxidative stress (OxS); ROS production rate by electron paramagnetic resonance (EPR) technique; and oxidative damage biomarkers (thiobarbituric acid reactive substances (TBARS) and protein carbonyls (PC)), electroneurography (ENG) and visual analogue scale. p < 0.05) at 30 and 60 days. ROS production rate up to \u221216%; TBARS (\u221231%), PC (\u221238%), and TAC up to +48%. Motor nerve conduction velocity in SPE and ulnar nerves (+22% and +16%) and sensor conduction velocity in sural and median nerves (+22% and +5%). Patients reported a general wellness sensation improvement (+35%) at 30 days: lower limb pain sensation (\u221240%) and upper limbs (\u221223%). Supplementation induced significant changes ( The results strongly indicate that an increased antioxidant capacity plays an important role in OxS, nerve conduction velocity, pain, and general wellness improvement. Nevertheless, the effects of the antioxidant compound were found positive up to 60 days. Then, a hormesis effect was observed. Novelty of the research would be a challenge for investigators to carefully address issues, including dose range factors, appropriate administration time, and targeting population to counteract possible \u201cboomerang effects.\u201d The great number of monitored parameters would firmly stress these conclusions. Type 2 diabetes mellitus (type 2 DM) is the most common form of diabetes, accounting for 90\u201395% of patients in developed countries: in 2030, up to 438 million people affected by type 2 DM have been estimated by recent studies .Type 2 DM is a metabolic disorder, formerly known as non-insulin-dependent diabetes mellitus or adult-onset diabetes, resulting from a combination of insulin resistance and inadequate insulin secretion .Long-term and serious complications of diabetes develop gradually; they can eventually be disabling or even life-threatening . One of 2\u2022\u2212), hydroxyl (HO\u2022), peroxyl (RO2\u2212\u2022), and hydroperoxyl (HRO2\u2212\u2022\u2212) and nonradical species, such as hydrogen peroxide (H2O2) and hydrochloric acid (HOCl) is also reported in the text. Sample size calculation to determine the minimum number of subjects for adequate study power was made by using the Freeware G\u2217Power software (http://www.psycho.uni-duesseldorf.de/abteilungen/aap/gpower3/). At 80% power, the calculated sample size was of 11 subjects, slightly lower than the subject's population recruited in the present study.All data were presented as mean\u2009\u00b1\u2009SD and were analysed using repeated Shapiro-Wilks 2, blood pressure, heart rate, and pharmacological therapy did not show any significant difference. At T0, the biochemical parameters and the haematological indices resulted in the reference ranges as follows: erythrocyte indices: RBCs: 4.63\u2009\u00b1\u20090.48 106/mm3, HGB: 14.04\u2009\u00b1\u20091.16\u2009g/dL, HCT: 41.46\u2009\u00b1\u20093.52%, MCV: 89.83\u2009\u00b1\u20092.60\u2009\u03bcm3, MCH: 30.44\u2009\u00b1\u20091.04\u2009pg, MCHC: 33.89\u2009\u00b1\u20090.47\u2009g/dL, and RDW: 13.78\u2009\u00b1\u20090.95%; leukocyte indices: WBCs: 6.58\u2009\u00b1\u20091.60 103/mm3, %LYM 27.65\u2009\u00b1\u20096.87, %MON 8.95\u2009\u00b1\u20092.62, %GRA 58.94\u2009\u00b1\u20098.88, #LYM: 1.89\u2009\u00b1\u20090.83 103/mm3, #MON: 0.56\u2009\u00b1\u20090.10 103/mm3, and #GRA 3.81\u2009\u00b1\u20090.80 103/mm3; platelet indices: PLT: 219.75\u2009\u00b1\u200951.01 103/mm3 and MPV: 9.50\u2009\u00b1\u20090.92\u2009\u03bcm3; S-lactate dehydrogenase ((LDH): 327.63\u2009\u00b1\u200977.55\u2009U/L); S-creatinine (0.86\u2009\u00b1\u20090.32\u2009\u03bcmol/L); C-reactive protein ((CRP): 0.23\u2009\u00b1\u20090.33\u2009mg/L). By contrast, haemoglobin A1c ((HbA1c): 6.93\u2009\u00b1\u20090.69%) and glycaemic index ((GI): 165.77\u2009\u00b1\u200932.34\u2009mg/dL) resulted strong increases with respect to the normal range.During the supplementation period of R(+)-thioctic acid and for the whole duration of the study, subjects' weight, BMI, SaOAll biochemical and haematological parameters, monitored at T3, T4, T6, and T8, did not show any statistically significant differences with respect to the basal T0 level.However, the glycaemic index followed a trend parallel to OxS, showing an 8% decrease at T4 and a 26% increase at T6, slowly returning to the basal level after the wash out time period.\u03bcmol\u00b7min\u22121) measured at the different times (T0\u2013T8) in type II DN patients' capillary blood and plasma. The ROS production rate data calculated at any time and the statistically significant differences between times of measurements are shown in Figures Capillary blood (A):T0 (2.14\u2009\u00b1\u20090.21) versus T1 (1.97\u2009\u00b1\u20090.25), T2 (1.77\u2009\u00b1\u20090.31), T3 (1.79\u2009\u00b1\u20090.21), and T6 (2.33\u2009\u00b1\u20090.28); T1 (1.97\u2009\u00b1\u20090.25) versus T6 (2.33\u2009\u00b1\u20090.28), T7 (2.19\u2009\u00b1\u20090.20), and T8 (2.16\u2009\u00b1\u20090.20); T2 (1.77\u2009\u00b1\u20090.31) versus T5 (2.24\u2009\u00b1\u20090.12) and T6 (2.33\u2009\u00b1\u20090.28); T3 (1.79\u2009\u00b1\u20090.21) versus T5 (2.24\u2009\u00b1\u20090.12), T6 (2.33\u2009\u00b1\u20090.28), T7 (2.19\u2009\u00b1\u20090.20), and T8 (2.16\u2009\u00b1\u20090.20).in Plasma (B):T0 (0.20\u2009\u00b1\u20090.03) versus T6 (0.27\u2009\u00b1\u20090.04); T2 (0.19\u2009\u00b1\u20090.03) versus T6 (0.27\u2009\u00b1\u20090.04); T3 (0.18\u2009\u00b1\u20090.03) versus T5 (0.22\u2009\u00b1\u20090.02), T6 (0.27\u2009\u00b1\u20090.04), and T7 (0.22\u2009\u00b1\u20090.02); T4 (0.22\u2009\u00b1\u20090.02) versus T6 (0.27\u2009\u00b1\u20090.04); T5 (0.22\u2009\u00b1\u20090.02) versus T6 (0.27\u2009\u00b1\u20090.04); T6 (0.27\u2009\u00b1\u20090.04) versus T7 (0.22\u2009\u00b1\u20090.02) and T8 (0.21\u2009\u00b1\u20090.02).The antioxidant supplementation of R(+)-thioctic acid induced EPR detectable changes in the ROS production levels (\u03bcM) and PC (nmol\u00b7mg\u22121 protein) concentration data measured at any time and the statistically significant differences between times of measurements are shown in Figures At the same time, the antioxidant supplementation induced changes in the investigated oxidative markers. TBARS (TBARS (A):T2 (14.37\u2009\u00b1\u20094.68) versus T6 (19.02\u2009\u00b1\u20095.60) and T7 (17.82\u2009\u00b1\u20094.93); T3 (12.38\u2009\u00b1\u20094.51) versus T6 (19.02\u2009\u00b1\u20095.60) and T7 (17.82\u2009\u00b1\u20094.93); T4 (11.57\u2009\u00b1\u20093.49) versus T6 (19.02\u2009\u00b1\u20095.60) and T7 (17.82\u2009\u00b1\u20094.93).PC (B):T0 (1.75\u2009\u00b1\u20090.62) versus T2 (1.39\u2009\u00b1\u20090.60), T3 (1.63\u2009\u00b1\u20090.46), T4 (1.08\u2009\u00b1\u20090.46), and T5 (1.56\u2009\u00b1\u20090.59); T2 (1.39\u2009\u00b1\u20090.60) versus T6 (1.99\u2009\u00b1\u20090.65), T7 (1.90\u2009\u00b1\u20090.60), and T8 (1.77\u2009\u00b1\u20090.59); T3 (1.63\u2009\u00b1\u20090.46) versus T6 (1.99\u2009\u00b1\u20090.65), T7 (1.90\u2009\u00b1\u20090.60), and T8 (1.77\u2009\u00b1\u20090.59); T4 (1.08\u2009\u00b1\u20090.46) versus T5 (1.56\u2009\u00b1\u20090.59), T6 (1.99\u2009\u00b1\u20090.65), T7 (1.90\u2009\u00b1\u20090.60), and T8 (1.77\u2009\u00b1\u20090.59); T5 (1.56\u2009\u00b1\u20090.59) versus T6 (1.99\u2009\u00b1\u20090.65).More specifically, the differences between brackets in the Figure are as follows:Finally, the antioxidant capacity in plasma, TAC (mM) concentration data measured at any time, and the statistically significant differences between times of measurements are reported in More specifically, the differences between brackets in the Figure are as follows:T0 (1.42\u2009\u00b1\u20090.38) versus T5 (2.21\u2009\u00b1\u20090.19) and T6 (2.36\u2009\u00b1\u20090.15); T2 (1.72\u2009\u00b1\u20090.17) versus T5 (2.21\u2009\u00b1\u20090.19) and T6 (2.36\u2009\u00b1\u20090.15); T3 (1.85\u2009\u00b1\u20090.18) versus T5 (2.21\u2009\u00b1\u20090.19) and T6 (2.36\u2009\u00b1\u20090.15); T4 (2.10\u2009\u00b1\u20090.19) versus T6 (2.36\u2009\u00b1\u20090.15) and T8 (1.59\u2009\u00b1\u20090.18); T5 (2.21\u2009\u00b1\u20090.19) versus T8 (1.59\u2009\u00b1\u20090.18); T6 (2.36\u2009\u00b1\u20090.15) versus T7 (1.80\u2009\u00b1\u20090.21) and T8 (1.59\u2009\u00b1\u20090.18).In the patient group examined in the present study, the four item symptom DNS score resulted as follows: neuropathic pain: 100% (12 patients); paraesthesia: 58% (7 patients); unsteadiness in walking: 8% (1 pz); numbness in the legs or feet: 0%. Therefore, a score (\u00b1SD) of 1.6\u2009\u00b1\u20090.65, positive for PNP, was calculated. ENG abnormalities were monitored in all patients. Changes in signal amplitude (mV) and distal latency (m/s) in sensory and motor nerves at T0, T3, T4, T6, and T8 were observed (data not shown).R(+)-thioctic acid supplementation induced changes in motor (MNCV m/s) and sensitive (SNCV m/s) conduction velocities see calculatMNCV (m/s) in ulnar nerve increased significantly at :T3 (53.62\u2009\u00b1\u20096.48) versus T0 (48.06\u2009\u00b1\u20095.58); T3 versus T6 (49.52\u2009\u00b1\u20094.98) and T8 (48.28\u2009\u00b1\u20095.36);superficial peroneal nerve (SPE) MNCV (m/s) increased significantly at :T3 (46.68\u2009\u00b1\u20096.42) and T4 (46.70\u2009\u00b1\u20096.43) versus T0 (38.30\u2009\u00b1\u20095.53).SNCV (m/s) in median wrist-elbow changed significantly at :T0 (52.38\u2009\u00b1\u20094.89) versus T3 (58.76\u2009\u00b1\u20093.24); T3 (58.76\u2009\u00b1\u20093.24) versus T6 (59.16\u2009\u00b1\u20093.90) and T8 (53.60\u2009\u00b1\u20095.96); T6 (59.16\u2009\u00b1\u20093.90) versus T8 (53.60\u2009\u00b1\u20095.96).SNCV (m/s) in sural nerve changed significantly at T0 (a), TBARS (b), and PC concentration data (c) and ROS production in plasma versus TBARS (d), PC (e), and glycaemic values (f) are also displayed.In particular, ROS production in capillary blood versus ROS in plasma . In particular, multiple sources of OxS in diabetes can be found, including nonenzymatic , enzymatic , and mitochondrial pathways , 38. Hyp\u03b1-lipoic acid, evening primrose oil (EPO), or sunflower oil was found to decrease plasma lipid levels and risk factors [\u03b1-lipoic acid on oxidative stress parameters are reported in the literature: increased super oxide dismutase (SOD) and subsequent reduction of H2O2 by catalase [Dietary supplementation with factors . The poscatalase , 42.2\u2212) levels [Direct evidence of OxS in diabetes was previously provided by the measurement of oxidative stress markers, such as plasma and urinary F2-isoprostane, as well as plasma and tissue nitrotyrosine and superoxide anion radicals (O\u2022) levels , 44.Starting from these considerations, the present study aimed at investigating the effects produced by antioxidant supplementation -thioctic acid, 1.6\u2009g/die) and after the wash out period (two months) on diabetic neuropathic patients throughout ROS production measurement by adopting an innovative EPR method.The study protocol was sketched in As observed in As a matter of fact, in all patients at T0 that is just before the antioxidant supply , a high degree of oxidative damage, as confirmed by the high ROS production rate concentration values, was measured in both capillary blood and plasma.Indeed, the acute effects, \u201cone shot\u201d of R(+)-thioctic acid supplementation in differently aged healthy subjects, were previously reported . A decreIndeed, the results of the present study conducted on diabetic patients, homogeneous for age and physical characteristics see and concp < 0.01) and kept decreasing at T3 and T4 of daily supplementation. According to the ROS decrease, also, TBARS and PC levels significantly (p < 0.01) decreased at T3 (\u221226 and \u221239%) and T4 (\u221231% and 38%), respectively increased as shown by the increase of ROS production in capillary blood and plasma , when compared to the basal level increased : the OxS values significantly , then returning slowly towards the baseline during the wash out and the periods without administration even if at a slightly higher level . This figure was found also for the ROS production and oxidative damage markers -thioctic acid on ROS production and oxidative damage markers, as well as on motor and sensory nerve conduction velocities see and 3. TThe role played by OxS in nerve damage has been extensively studied in experiments carried out in an animal model and moreDiabetic neuropathies are diabetes mellitus-associated disorders probably ascribable to different causes; some of them are not yet well elucidated and in particular how prolonged exposure to high blood glucose levels can produce nerve damage.\u03b1-lipoic acid on nerve functions. In fact, \u03b1-lipoic acid has been shown to improve motor nerve conduction velocity in experimental diabetic neuropathy and to protect peripheral nerves from ischemia in rats [Motor nerve (MNCV) and sensory nerve (SNCV) conduction velocities resulted in the primary endpoints for studying the therapeutic effectiveness of in rats .Evidence related to changes in nerve conduction velocity after thioctic acid administration was observed primarily in experimental models \u201360.The conduction velocities (m/s) of the sensory component of ulnar and sural nerves in diabetic subjects affected with neuropathy were previously found significantly lower than those measured in diabetics without neuropathy as well as nondiabetic subjects . In well\u03b1-lipoic acid (600\u2009mg/die). This molecule is known to interact with many glucose-converting enzymes, fatty acids, and other energy sources in ATP. In the present study, nerve conduction significantly increased at T3 and T4 (30\u201360 supplementation days): MNCV: +16-17% in ulnar and external sciatic-popliteal (SPE) nerves; SNCV: +12-13% in median and about +16% in sural nerves.Again, the literature data reportedIndeed, in human studies to date, a great number of studies targeting \u201cthioctic acid\u201d and \u201cdiabetes mellitus\u201d have been published, from which only a few randomized control trials in humans are distinguished in terms of DPN treatment with thioctic acid. We are well aware that nerve conduction velocity cannot be used to monitor changes in oxidative stress; nonetheless, the data of the present study suggested us that the hormesis effect described above could be hypothesized for MNCV and SNCV too: the beneficial effects on nerve conduction were lost after long antioxidant administration time period (T6 and over), almost returning to the pretreatment (T0) conduction velocity values see .VAS is a suitable method to measure pain/pain relief and general wellness. In the present study, in all patients, a regression of the subjective pain sensation as well as a general wellness improvement was reported under R(+)-thioctic acid treatment mainly at T3 and T4 , followed by an improvement of about the 50% after 30\u201360\u2009days of treatment , then returning to the basal level after the wash out period. At the same time, at T3-T4, the general wellness sensation was found to improve at about +35% as well.Side effects, like stomachache and nausea, were reported, but mild or moderate, since no patients interrupted the supplementation for this reason.Side effects as a consequence of alpha-lipoic acid oral administration were previously reported by Ziegler et al. . DespiteA significantly higher dose protocol was followed in the present study (1.6\u2009g/die): patients referred stomachache and nausea mainly at T2-T3 and T5-T6, the symptoms returning to normal after the wash out period see .Nevertheless, during the supplementation time course, patients reported an improvement of the general wellness sensation mainly at T3-T4: in particular, pain at the lower limbs and feet duration of the treatment, no significant differences were found in the haematological parameters all over the supplementation duration.p < 0.01) linear correlation was found at every time between the ROS production mean values in capillary blood and ROS production in plasma (R2 = 0.76), TBARS (R2 = 0.60), and PC (R2 = 0.79) relationships were found at every time between ROS production mean values in plasma and TBARS (R2 = 0.58), PC (R2 = 0.66), and glycaemic levels (R2 = 0.54) was large enough to ensure an adequate study power, as determined by the Freeware G\u2217Power software (see paragraph 2.7). In fact, as already pointed out, at 80% power, the calculated sample size was 11 subjects. However, the population recruited for the study is certainly well below the number of participants standardly recruited for a clinical trial. As a matter of fact, the present study has to be considered as a pilot test. The research concerns the use of an antioxidant substance, the R(+)-thioctic acid, whose acute effects were previously tested on healthy subjects by some of us. In type II diabetic patients, the pilot test study consisted in a very long administration time and the wash out period. Despite the long administration time, none of the patients interrupted the trail so that the results could be obtained at each time from the whole subject sample. It is worth noting the great number of monitored parameters joining oxidative stress by using innovative instruments (EPR) applied by a mini-invasive technique and electroneurography investigations, together with the homogeneity of the assessed results, supported and strengthened by the significance of the correlations -thioctic acid 1.6\u2009mg administration was shown to produce EPR detectable changes in ROS formation as measured in capillary blood and plasma and related oxidative damage parameters with consequent positive effects on nerve conduction velocity too. The findings of the present study strongly indicated that an increased antioxidant capacity plays an important role in the improvement of OxS, nerve conduction velocity, pain, and general wellness. Nevertheless, at the same time, the results of the study showed that the safety effects of an exogenous antioxidant compound were positive up to 60 administration days. After this period, a hormesis effect was observed. Therefore, the novelty of the conducted research would be a challenge for investigators to carefully address issues, including dose range factors, as well as appropriate administration time and targeting population to counteract possible \u201cboomerang effects\u201d of the treatment. The great number of parameters monitored in this study would firmly stress these conclusions."} +{"text": "Scientific Reports6: Article number: 3157410.1038/srep31574; published online: 08172016; updated: 02172017\u20134\u2009\u03bc W cm\u22122, at 4.5\u2009cm) illuminated the surface of the polymer samples\u201d.This Article contains an error in the Methods section. \u201cTwo 30\u2009W UVC lamps illuminated the surface of the polymer samples\u201d.\u201cTwo 30\u2009W UVC lamps (253.7\u2009nm, irradiance of 0.01\u2009W\u2009cm"} +{"text": "I atom in the mol\u00adecular structure of the title compound exhibits a distorted tetra\u00adhedral coordination sphere, defined by two N atoms of the chelating ligand, one N atom of the aceto\u00adnitrile ligand and one iodide ligand.The Cu 2H3N)(C26H24N2)], the CuI ion has a distorted tetra\u00adhedral coordination environment, defined by two N atoms of the chelating 2--6-[imino]\u00adpyridine ligand, one N atom of an aceto\u00adnitrile ligand and one iodide ligand. Within the complex, there are weak intra\u00admolecular C\u2014H\u22efN hydrogen bonds, while weak inter\u00admolecular C\u2014H\u22efI inter\u00adactions between complex mol\u00adecules, help to facilitate a three-dimensional network.In the mononuclear title complex, [CuI(C The two N atoms of the bidentate ligand chelate to CuI with similar Cu\u2014N bond lengths . A comparable N,N\u2032-binding has been observed in related structures with bis\u00ad[2-(2-pyrid\u00adyl)eth\u00adyl]amine ligands \u2005\u00c5. The N2\u2014Cu1\u2014N1 bite angle of the chelating ligand is 78.86\u2005(18)\u00b0, while the N3\u2014Cu\u2014I angle between the monodentate aceto\u00adnitrile and iodide ligands is closer to tetra\u00adhedral, 112.74\u2005(15)\u00b0. The naphthyl ring system is inclined by 58.20\u2005(17)\u00b0 to the central N=C(CH3)\u2014pyridine moiety, whereas the tri\u00admethyl\u00adphenyl ring is almost perpendicular to the latter, at 84.8\u2005(3)\u00b0. Within the complex, an intra\u00admolecular C\u2014H\u22efN hydrogen-bonding inter\u00adaction is present, stabil\u00adizing the mol\u00adecular conformation \u2005\u00c5, 152\u00b0; symmetry code: (i) x, y\u00a0\u2212\u00a01, z] and a H atom of the aceto\u00adnitrile methyl group link the complex mol\u00adecules, forming a three-dimensional network was synthesized according to a modified literature procedure in 5\u2005ml of aceto\u00adnitrile. The mixture was stirred at room temperature for 24\u2005h before evaporating the volatiles. The residue was extracted with n-hexane (5 \u00d7 3\u2005ml). The extracts were combined and the solvent removed under reduced pressure to give a red solid which was recrystallized from aceto\u00adnitrile solution. Yield: 54%. M.p. >253\u2005K (decomp). 1H NMR : \u03b4 1.88 , 1.97 , 2.16 , 2.20 , 6.84 , 7.39 , 7.45 , 7.51 , 7.73 , 7.81 , 7.87 , 8.04 . IR \u03bdmax (solid)/cm\u22121 1620 (C=Nimine), 1555 (C=Npy). ESI MS: m/z 428 [M\u2013I\u2013MeCN]+.A solution of 0.0262\u2005g of CuI (0.137\u2005mmol) in 5\u2005ml of aceto\u00adnitrile was mixed with a solution of 0.05\u2005g of Uiso(H) = 1.2Ueq(C) for H atoms on Csp2 and 0.98\u2005\u00c5 with Uiso(H) = 1.5Ueq(C) for H atoms on Csp3.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016018685/wm5341sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016018685/wm5341Isup2.hklStructure factors: contains datablock(s) I. DOI: 1518571CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II ion is completed by the ligation of two N atoms of two piperazinium ions. In the crystal, a network of N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds leads to the formation of a three-dimensional supra\u00admolecular structure.In the title compound, two thio\u00adsulfate ions coordinate to the zinc(II) ion through the terminal S atoms. The tetra\u00adhedral coordination around the Zn 4H11N2)2(S2O3)2]\u00b72H2O, two thio\u00adsulfate ions coordinate to the zinc(II) atom through the terminal S atoms. The tetra\u00adhedral coordination around the ZnII ion is completed by ligating to two N atoms of two piperazinium ions. The remaining two N atoms of the piperazinium ions are diprotonated and do not coordinate to the metal centre. In the crystal, however, they are involved in N\u2014H\u22efOwater and N\u2014H\u22efOsulfato hydrogen bonds. Together, a series of N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, involving the O atoms of the thio\u00adsulfate ions and the water mol\u00adecules as acceptors and the hydrogen atoms of the piperazinium ions and the water mol\u00adecules as donors, form a three-dimensional supra\u00admolecuar structure. Within this framework there are a number of intra- and inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efS contacts present.In the title compound, [Zn(C Recently, Natarajan and co-workers and two nitro\u00adgen atoms from the piperazinium ions (N1 and N3), in an approximately tetra\u00adhedral geometry . The Zn\u2014S bond lengths are 2.2927\u2005(4)\u2005\u00c5 for Zn1\u2014S1 and 2.3324\u2005(4)\u2005\u00c5 for Zn1\u2014S3. The Zn\u2014N bond lengths are 2.0879\u2005(13)\u2005\u00c5 for Zn1\u2014N1 and 2.0727\u2005(12)\u2005\u00c5 for Zn1\u2014N3. The N/S\u2014Zn1\u2014S/N bond angles lie in the range 101.24\u2005(4) to 116.79\u2005(2)\u00b0, confirming the tetra\u00adhedral nature of the zinc ions. Within the two thio\u00adsulfate ligands, the S\u2014S bond lengths are 2.0511\u2005(5)\u2005\u00c5 for S1\u2014S2 and 2.0332\u2005(5)\u2005\u00c5 for S3\u2014S4. The S\u2014O bond lengths vary from 1.4437\u2005(14) to 1.4623\u2005(13)\u2005\u00c5, while the O\u2014S\u2014O angles vary from 104.53\u2005(5) to 112.85\u2005(10)\u00b0, which is indicative of a fairly regular tetra\u00adhedral arrangement. In the mol\u00adecular unit, the two thio\u00adsulfate units are bonded to the zinc(II) ion only through the terminal S atoms, and the oxygen atoms are uncoordinated. In addition, only one nitro\u00adgen atom of each piperazinium ion is bonded to the zinc(II) ion, the second being diprotonated in each case.The mol\u00adecular structure of the title compound is illustrated in Fig.\u00a01The supra\u00admolecular architecture Fig.\u00a02 arises fet al., 2016et al. zinc(II) hemihydrate (CSD refcode: WEHTOT). The thio\u00adsulfate ligand is coordinated to the zinc ions through S and O atoms, forming octa\u00adhedral zinc centres. In addition, a zinc\u2013thio\u00adsulfate complex containing both one-dimensional cationic and anionic chains has been reported by the same authors, viz. catena-[tetra\u00adaqua\u00adzinc(II) bis\u00adbis\u00ad(thio\u00adsulfato-\u03baS)dizinc(II) dihydrate] (IJUWER), catena-[(\u03bc-thio\u00adsulfato)\u00adzinc] (IJU\u00adWIV), and catena-[bis\u00ad\u00adbis\u00ad(\u03bc-thio\u00adsulfato)\u00addizinc dihydrate] (IJUWOB) and catena-[bis\u00ad\u00adbis\u00ad(\u03bc-thio\u00adsulfato)\u00addizinc (\u03bc-thio\u00adsulfato)\u00adzinc trihydrate] (IJUWUH).Karthik & Natarajan 2016 have recet al., 1996et al., 2001et al., 2004A number of mol\u00adecular cadmium\u2013thio\u00adsulfate and manganese\u2013thio\u00adsulfate structures have been reported by Baggio and co-workers (thio\u00adsulfato)cadmium (CSD refcode: ORUJOC), which was reported recently, was isolated in the presence of the aliphatic amine 1,3-di\u00adamino\u00adpropane 2\u00b76H2O was dissolved in 5\u2005ml distilled water. Then (NH4)2S2O3 was added to the solution, which was stirred for 15\u2005min. Piperazine was dissolved separately in distilled water (5\u2005ml) and the solution poured into the initial reaction mixture until the pH was 8. The resulting solution was left undisturbed and after 1 week, colourless block-shaped crystals were obtained. The product was filtered and washed with cold water. The yield was approximately 85% based on Zn metal. Elemental analysis calculated for C8H26N4O8S4Zn: C 19.20, H 5.24, N 11.20%; found: C 19.27, H 5.29, N 11.16%.Zn = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018000555/cq2022sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989018000555/cq2022Isup2.hklStructure factors: contains datablock(s) I. DOI: 1571150CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-017-03589-w, published online 12 June 2017Correction to: The original version of this Article contained a typographical error in the Abstract.9/L)\u2009\u00d7\u2009AST(U/L).\u201d\u201cThe formula of the mFIB-4 index is 10\u2009\u00d7\u2009Age(years)\u2009\u00d7\u2009AST(U/L)/Platelet count(10now reads:9/L)\u2009\u00d7\u2009ALT(U/L).\u201d\u201cThe formula of the mFIB-4 index is 10\u2009\u00d7\u2009Age(years)\u2009\u00d7\u2009AST(U/L)/Platelet count(10This has now been corrected in the PDF and HTML versions of the Article."} +{"text": "Crystal structures of hydrogen-bonded 1:2 dihydrate compounds of chloranilic acid with 2-carb\u00adoxy\u00adpyridine (I) and 2-carb\u00adoxy\u00adquinoline (II) have been determined at 180 and 200\u2005K, respectively. The base mol\u00adecule in (I) is disordered over cationic and twitterionic states, while that in (II) is in a twitterionic form. In each crystal, the three components are linked by O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming a layer structure. 6H5.5NO20.5+\u00b7C6HCl2O4\u2212\u00b72H2O, (I), has been determined at 180\u2005K, and the structure of the 1:2 dihydrate compound of chloranilic acid with 2-carb\u00adoxy\u00adquinoline , namely, 2C10H7NO2\u00b7C6H2Cl2O4\u00b72H2O, (II), has been redetermined at 200\u2005K. This determination presents a higher precision crystal structure than the previously published structure [Marfo-Owusu & Thompson was analysed as a disordered structure over two states, viz. salt and co-crystal. The salt is bis\u00ad(2-carb\u00adoxy\u00adpyridinium) chloranilate dihydrate, 2C6H6NO2+\u00b7C6Cl2O42\u2212\u00b72H2O, and the co-crystal is bis\u00ad(pyridinium-2-carboxyl\u00adate) chloranilic acid dihydrate, 2C6H5NO2\u00b7C6H2Cl2O4\u00b72H2O, including zwitterionic 2-carb\u00adoxy\u00adpyridine. In both salt and co-crystal, the water mol\u00adecule links the chloranilic acid and 2-carb\u00adoxy\u00adpyridine mol\u00adecules through O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds. The 2-carb\u00adoxy\u00adpyridine mol\u00adecules are connected into a head-to-head inversion dimer by a short O\u2014H\u22efO hydrogen bond, in which the H atom is disordered over two positions. Compound (II) is a 1:2 dihydrate co-crystal of chloranilic acid and zwitterionic 2-carb\u00adoxy\u00adquinoline. The water mol\u00adecule links the chloranilic acid and 2-carb\u00adoxy\u00adquinoline mol\u00adecules through O\u2014H\u22efO hydrogen bonds. The 2-carb\u00adoxy\u00adquinoline mol\u00adecules are connected into a head-to-tail inversion dimer by a pair of N\u2014H\u22efO hydrogen bonds.The crystal structure of the 1:2 dihydrate compound of chloranilic acid with 2-carb\u00adoxy\u00adpyridine , namely, 2Cpson 2014. X-ray S The pyridine rings are also stacked along the b axis through a \u03c0\u2013\u03c0 inter\u00adaction [centroid\u2013centroid distance = 3.6851\u2005(7)\u2005\u00c5 and inter\u00adplanar spacing = 3.4787\u2005(5)\u2005\u00c5]. Between the layers, a short Cl\u22efCl contact is observed [Cl1\u22efCl1v = 3.3717\u2005(5)\u2005\u00c5; symmetry code: (v) \u2212x\u00a0+\u00a01, y\u00a0\u2212\u00a0z\u00a0+\u00a0In the crystal of compound (I)ne Fig.\u00a04. In the via a pair of N\u2014H\u22efO hydrogen bonds \u2005\u00c5. The water mol\u00adecule links the stacked base mol\u00adecules related by translation along a via O\u2014H\u22efO hydrogen bonds [O5\u2014H3\u22efO4 and O5\u2014H4\u22efO4ii; Table\u00a02via O\u2014H\u22efO hydrogen bonds, forming a layer structure parallel to et al., 2016A search of the Cambridge Structural Database Uiso(H) = 1.5Ueq(O). For the water H atoms, distant restraints of H\u22efH = 1.37\u2005(4)\u2005\u00c5 were also applied. C-bound H atoms were positioned geometrically (C\u2014H = 0.95\u2005\u00c5) and were treated as riding with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a03Uiso(H) = 1.2Ueq(C).All H atoms in compound (II)10.1107/S2056989017015997/lh5860sup1.cifCrystal structure: contains datablock(s) General, I, II. DOI: 10.1107/S2056989017015997/lh5860Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017015997/lh5860IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989017015997/lh5860IIsup4.cmlSupporting information file. DOI: 1583721, 1583720CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "DNGR\u20101 is receptor expressed by certain dendritic cell (DC) subsets and by DC precursors in mouse. It possesses a C\u2010type lectin\u2010like domain (CTLD) followed by a poorly characterized neck region coupled to a transmembrane region and short intracellular tail. The CTLD of DNGR\u20101 binds F\u2010actin exposed by dead cell corpses and causes the receptor to signal and potentiate cross\u2010presentation of dead cell\u2010associated antigens by DCs. Here, we describe a conformational change that occurs in the neck region of DNGR\u20101 in a pH\u2010 and ionic strength\u2010dependent manner and that controls cross\u2010presentation of dead cell\u2010associated antigens. We identify residues in the neck region that, when mutated, lock DNGR\u20101 in one of the two conformational states to potentiate cross\u2010presentation. In contrast, we show that chimeric proteins in which the neck region of DNGR\u20101 is replaced by that of unrelated C\u2010type lectin receptors fail to promote cross\u2010presentation. Our results suggest that the neck region of DNGR\u20101 is an integral receptor component that senses receptor progression through the endocytic pathway and has evolved to maximize extraction of antigens from cell corpses, coupling DNGR\u20101 function to its cellular localization. Bufferlong mouse DNGR\u20101 ECD to mildly denaturing (Laemmli buffer) or strongly denaturing (8\u00a0M urea) conditions in the presence of DTT and analyzed the samples by SDS\u2013PAGE and Western blot. As predicted, strongly denaturing conditions almost completely abolished the ability of DNGR\u20101 to resist reduction, while under weakly denaturing conditions the reduction\u2010insensitive dimers could be observed. In the absence of reducing agents, the protein maintained its dimeric status regardless of the denaturing conditions to induce the reduction\u2010resistant state, dialyzed the protein back into PBS, and tested its reduction sensitivity. As expected, we observed reduction\u2010resistant dimers under mildly acidic conditions , all the proteins were readily reducible while, when subjected to weakly acidic conditions (pH 6.1), they showed increased resistance to reduction Fig\u00a0B. These We expressed a chimeric ECD, in which the neck region of DNGR\u20101 was fused to the CTLD of another C\u2010type lectin receptor, dectin\u20101 Fig\u00a0A. Dectinlong mouse and human DNGR\u20101 isoforms under neutral (PBS) or mildly acidic (MES pH 6.1) conditions, and observed almost perfect overlap between the two . Far\u2010UV CD spectra reflect the secondary structure content of the protein while near\u2010UV CD signals derive from the three aromatic residues and, in some cases, from disulfide bonds, and reflect protein tertiary structure conditions , suggesting that the part of the neck region involved in the process is conserved between the two. In order to pinpoint its location, we genetically removed overlapping blocks of 10\u201311 amino acids from the conserved part of the neck [K57\u2013L66 (\u03941), L64\u2013I73 (\u03942), L72\u2013L82 (\u03943), N81\u2013T90 (\u03944), R87\u2013A96 (\u03945), and Q95\u2013S104 (\u03946) resulted in an ECD protein that behaved like the \u03944 mutant in that it formed type\u20102 dimers in neutral conditions overnight and measured the extent of T\u2010cell reactivation by assessing the amount of IFN\u2010\u03b3 that accumulated in the culture medium. Consistent with previous data . pH\u2010induced conformational changes have been reported for other proteins and are often mediated by protonation of one or more histidines stabilizing forces in the same range as electrostatic repulsion, they can contribute to, and perhaps even be sufficient for, pH\u2010sensitive conformational rearrangements . Thus, other parts of DNGR\u20101 are also involved in determining the ability of the receptor to promote cross\u2010presentation of dead cell\u2010associated antigens.long mouse, K57\u2013I238 for short mouse, and K57\u2013V241 for the human isoform) or mouse dectin\u20101 (R71\u2013L244) was inserted into the p3xFLAG\u2010CMV\u20109 expression vector (Sigma\u2010Aldrich) and all the constructs were verified by sequencing. The proteins were expressed by transient transfection in 239F cells, as described previously and following primers: C94S: 5\u2032\u2010CCATGCATGATCCAATTAGGAAGGCATGTTTCCTTGCTGTCCAGGGTCTG\u20103\u2032, \u03941: 5\u2032\u2010CAAGGATGACGATGACAAGCTTGCGGCCGCGGAGCAGCAGGAAAGACTCATCCAACAGGAC\u20103\u2032, \u03942: 5\u2032\u2010CGGCCGCGAAGTTCTTCCAGGTATCCTCTCAACAGGACACAGCATTGGTGAACCTTACAC\u20103\u2032, \u03943: 5\u2032\u2010CCTCTCTTGTCTTGGAGCAGCAGGAAAGAACACAGTGGCAGAGGAAATACACACTGGAATACTGCC\u20103\u2032, \u03943A: 5\u2032\u2010GGTATCCTCTCTTGTCTTGGAGCAGCAGGAAAGAGCCGCCGCCGCCGCCGCCGCCGCCGCCGCCGCCACACAGTGGCAGAGGAAATACACACTGGAATACTGCCAAGCC\u20103\u2032, \u03944: 5\u2032\u2010GGAAAGACTCATCCAACAGGACACAGCATTGGTGCTGGAATACTGCCAAGCCTTACTGCAGAGATCTCTCC\u20103\u2032, \u03944A: 5\u2032\u2010GCAGGAAAGACTCATCCAACAGGACACAGCATTGGTGGCCGCCGCCGCCGCCGCCGCCGCCGCCGCCCTGGAATACTGCCAAGCCTTACTGCAGAG\u20103\u2032, \u03945: 5\u2032\u2010GACACAGCATTGGTGAACCTTACACAGTGGCAGTTACTGCAGAGATCTCTCCATTCAGGCAC\u20103\u2032, \u03946: 5\u2032\u2010CAGTGGCAGAGGAAATACACACTGGAATACTGCGGCACAGATGCTTCTACTGGACCAG\u20103\u2032, \u03946A: 5\u2032\u2010GTGAACCTTACACAGTGGCAGAGGAAATACACACTGGAATACTGCGCCGCCGCCGCCGCCGCCGCCGCCGCCGCCGGCACAGATGCTTCTACTGGACCAGTTCTTCTGAC\u20103\u2032.long mouse DNGR\u20101 (K57\u2013T130) with 5\u2032 and 3\u2032 sequences complementary to regions of the p3xFLAG\u2010CMV\u20109 plasmid and dectin\u20101, respectively, was amplified in a PCR with Phusion Hot Start II polymerase (Thermo Scientific), using the following primers: 5\u2032\u2010GCTGGGTGCCCTAGCATTTTGGAAGTTCTTCCAGGTATCCTCTCTTG\u20103\u2032 and 5\u2032\u2010CCATGCATGATCCAATTAGGAAGGCATGTTTCCTTGCTGTCCAGGGTCTG\u20103\u2032 and a DNGR\u20101\u2010coding plasmid as a template, and then used in a modified reaction with QuikChange Lightning kit (Agilent Technologies) as per manufacturer's instructions with p3xFLAG\u2010CMV\u20109 plasmid containing dectin\u20101 ECD sequence as a template. For generation of chimeric DNGR\u20101 proteins with the neck region of CD69 or Ly49, \u201cmegaprimers\u201d covering the entire neck region of appropriate proteins were generated using the following primers: 5\u2032\u2010GTTAGCAACGTCCATTTTCTTGGGCATCGGCAAGTACAATTGCCCAGGCTTG\u20103\u2032 and 5\u2032\u2010GTGGACAAGGGCTGCAGTCACTACCAGCAACATGGTGGTCAGATG\u20103\u2032 for CD69 and 5\u2032\u2010GTTAGCAACGTCCATTTTCTTGGGCATCAACATTTTTCAGAATAGTCAACAAAATCATGAACTGCAGG\u20103\u2032 and 5\u2032\u2010GTGGACAAGGGCTGCAGTCACTACCTTCAAAACCTCTGCCTGTGTGCTGTGAGG\u20103\u2032 for Ly49. cDNA generated from total splenic mRNA of a C57BL/6 and a CBA/J mouse, respectively, was used as templates. \u201cMegaprimers\u201d were then used in a modified reaction with QuikChange Lightning kit (Agilent Technologies) as per manufacturer's instructions with pFB\u2010IRES GFP plasmid containing the entire sequence of DNGR\u20101 as a template.For the chimeric DNGR\u20101:dectin\u20101 protein, a \u201cmegaprimer\u201d covering the whole neck region of Samples for SDS\u2013PAGE were prepared in 6\u00d7 Laemmli buffer (60% (v/v) glycerol, 150\u00a0mg/ml SDS, 0.75\u00a0mg/ml bromophenol blue in 75\u00a0mM Tris\u2013HCl pH 6.8) under reducing (+100\u00a0mM DTT) or non\u2010reducing conditions unless otherwise stated. Separation was carried out using 4\u201320% Mini\u2010PROTEAN TGX precast gels (Bio\u2010Rad). Proteins were transferred onto Immobilon P membrane (Merck Millipore) using wet transfer, the membrane was blocked in 5% milk in PBS + 0.05% Tween\u201020, and ECD proteins were detected using HRP\u2010conjugated M2 anti\u2010FLAG antibody (Sigma\u2010Aldrich). Full\u2010length DNGR\u20101 was detected using anti\u2010DNGR\u20101 antibody (clone 397) followed by HRP\u2010conjugated polyclonal anti\u2010rat antibody (IgG (H+L); Stratatech).long DNGR\u20101 ECD was concentrated to 7\u00a0mg/ml and, immediately before analysis, diluted into PBS or 10\u00a0mM MES pH 6.1 to final concentration 170\u00a0\u03bcg/ml. Human DNGR\u20101 ECD was concentrated to 1.4\u00a0mg/ml and immediately before analysis diluted into PBS or 10\u00a0mM MES pH 6.1 to final concentration 140\u00a0\u03bcg/ml. Far\u2010UV CD spectra (260\u2013195\u00a0nm) were recorded at 20\u00b0C in 1\u00a0mm fused silica cuvettes using a Jasco J\u2010815 spectropolarimeter fitted with a cell holder temperature controlled by a CDF\u2010426S Peltier unit. The spectra were typically recorded with 0.1\u2010nm resolution and baseline corrected by subtraction of the appropriate buffer spectrum. CD intensities are presented as the CD absorption coefficient calculated on a mean residue weight basis (\u0394\u03b5MRW). Secondary structure content was estimated using methods originally described by Sreerama and Woody were recorded at 20\u00b0C in 10\u00a0mm fused silica cuvettes. Near\u2010UV CD intensities are presented on a molecular weight basis (\u0394\u03b5M).For far\u2010UV CD, mouse et\u00a0al, et\u00a0al, 2) and polymerized in F\u2010buffer . Polymerized actin was stabilized with 5\u00a0\u03bcM phalloidin (Life Technologies) and spotted onto nitrocellulose membrane (Whatman) in twofold dilution series. The membrane was blocked in 5% milk in PBS + 0.05% Tween\u201020 overnight and incubated with protein supernatants diluted into appropriate buffers at equal concentrations. Binding of all proteins was detected using HRP\u2010conjugated M2 anti\u2010FLAG antibody (Sigma\u2010Aldrich) and revealed using SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific) or Luminata Forte Western HRP Substrate (Merck Millipore).Dot blot assay was performed as described previously supplemented with \u03b2\u2010ME and 10% heat\u2010inactivated fetal calf serum (FCS) at 37\u00b0C and 5% CO2. 293F cells were grown in FreeStyle 293 Expression Medium (Gibco) at 37\u00b0C, 8% CO2, and with constant shaking on an orbital shaker at 120\u00a0rpm. 293FT cells were grown in DMEM medium (Gibco) supplemented with 10% heat\u2010inactivated fetal calf serum (FCS) at 37\u00b0C and 10% CO2.Phoenix, B3Z\u2010Syk , VSV\u2010G envelope protein\u2010coding plasmid, and a pFB plasmid coding for the desired protein. On days 1, 2, and 3 post\u2010transfection, the pseudotyped virus\u2010containing culture medium was recovered, filtered, supplemented with 8\u00a0\u03bcg/ml polybrene (Sigma\u2010Aldrich), and immediately applied to target cells. The plate was centrifuged for 90\u00a0min at 2,500\u00a0\u00d7\u00a0g at room temperature and left in the incubator for a further 90\u00a0min. After the incubation, the medium was exchanged for fresh complete IMDM medium. On day 5, selection medium containing 0.5\u00a0\u03bcg/ml puromycin was applied. Surviving cells after 1\u00a0week of selection were expanded and transduced in an identical manner as described above with lentivirus generated in 293FT cells transfected with a mixture of Lipofectamine 2000, psPax2, pMD2.G, and pLX\u2010sgRNA plasmids. Successfully transduced cells were selected in medium containing 5\u00a0\u03bcg/ml blasticidin. Surviving cells after 1\u00a0week of selection were expanded and Cas\u20109 expression was induced by addition of 1\u00a0\u03bcg/ml doxycycline to the culture medium for 3\u00a0weeks. Cells that lost DNGR\u20101 expression were then purified by FACS sorting. The sequence of sgRNA used was as follows: CTGAACATTTGCTAGGGGAT. pCW\u2010Cas9 and pLX\u2010sgRNA plasmids were a gift from Eric Lander & David Sabatini (Addgene plasmids #50661 and #50662), pMD2.G and psPAX2 plasmids were a gift from Didier Trono (Addgene plasmids #12259 and #12260).293FT cells were transfected by a mixture of Lipofectamine 2000 (Thermo Fisher Scientific), psPax2, pMD2.G, and pCW\u2010Cas9 plasmids. Lentivirus\u2010containing culture medium was recovered on days 1, 2, and 3 post\u2010transfection, filtered, diluted 1:9, supplemented with 8\u00a0\u03bcg/ml polybrene (Sigma\u2010Aldrich), and immediately applied to MuTu cells. The plate was centrifuged for 90\u00a0min at 2,500\u00a0\u00d7\u00a0et\u00a0al, 2 for 45\u00a0min. After the incubation, the cells were harvested, washed in ice\u2010cold PBS + 5\u00a0mM EDTA, and fixed in 4% PFA. After fixation, the cells were washed in FACS buffer and surface\u2010stained with PE\u2010conjugated anti\u2010DNGR\u20101 antibody (clone 1F6). Staining was analyzed using LSR Fortessa Flow cytometer (BD Biosciences). Data analysis was performed in FlowJo 9.8.5 software (TreeStar).Internalization assay was described previously 293T cells at the indicated ratios, or in wells coated with anti\u2010DNGR\u20101 antibody (clone 7H11) or in medium alone in a 96\u2010well plate. LacZ activity after incubation overnight at 37\u00b0C and 5% CO2 was determined by lysing the cells in CPRG (Roche) containing buffer and measuring OD595 with OD655 as a reference at multiple time points.B3Z\u2010Syk reporter assay was described previously and left overnight to undergo secondary necrosis. Cell corpses were harvested, washed once in full RPMI1640 medium, and plated at indicated ratios with 6\u00a0\u00d7\u00a0104 MuTu cells per well in a U\u2010bottom 96\u2010well plate. After a 4\u2010h incubation at 37\u00b0C and 5% CO2, 3\u00a0\u00d7\u00a0104 pre\u2010activated OT\u2010I T cells (see below) were added to each well and incubated at 37\u00b0C and 5% CO2 overnight. Next morning, the plate was freeze\u2013thawed once, and total amount of IFN\u2010\u03b3 in the supernatants was determined by ELISA.Ovalbumin\u2010expressing mouse embryonic fibroblasts (bm1 T OVA MEFs; Sancho \u2212/\u2212 mice were bred at The Francis Crick Institute under specific pathogen\u2010free conditions in accordance with the national and institutional guidelines for animal care and approval by The Francis Crick Institute Animal Ethics Committee and by the Home Office, UK. Spleen and lymph nodes from one OT\u2010I Rag1\u2212/\u2212 mouse were homogenized into a single cell suspension. Cells were plated at 1\u00a0\u00d7\u00a0105 per ml in complete RPMI1640 medium supplemented with 0.1\u00a0nM SIINFEKL peptide. On day 3 of culture, IL\u20102 was added to a final concentration of 25\u00a0U/ml. CD8+ T cells were MACS\u2010enriched on day 5 and used immediately.OT\u2010I Rag1PH designed the study, performed and analyzed experiments, and wrote the manuscript. OS provided technical assistance, contributed to the design of experiments, and performed some of the experiments shown in Fig\u00a0The authors declare that they have no conflict of interest.Review Process FileClick here for additional data file.Source Data for Figure\u00a01Click here for additional data file.Source Data for Figure\u00a02Click here for additional data file.Source Data for Figure\u00a03Click here for additional data file.Source Data for Figure\u00a04Click here for additional data file.Source Data for Figure\u00a06Click here for additional data file."} +{"text": "The title structure consists of 3-amino\u00adpyridinium cations and 1\u2032-carb\u00adoxy\u00adferrocene-1-carboxyl\u00adate monoanions held together by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds. 5H7N2)[Fe(C6H4O2)(C6H5O2)], consists of 3-amino\u00adpyridinium cations and 1\u2032-carb\u00adoxy\u00adferrocene-1-carboxyl\u00adate monoanions. The ferrocenyl moiety of the anion adopts a typical sandwich structure, with Fe\u2014C distances in the range 2.0270\u2005(15)\u20132.0568\u2005(17)\u2005\u00c5. The anion possesses an eclipsed conformation, with the torsion angle \u03c6 (Csubst\u2014Cpcent\u2014Cpcent\u2014 Csubst) equal to 66.0\u00b0. The conformations of other 1\u2032-carb\u00adoxy\u00adferrocene-1-carboxyl\u00adate monoanions are compared and analyzed on the basis of literature data.The structure of the title salt, (C The C16\u2014O bond lengths within the carboxyl\u00adate anion are almost equal [1.2604\u2005(19) and 1.2636\u2005(19)\u2005\u00c5], whereas, in contrast, they differ greatly within the carb\u00adoxy\u00adlic acid group, with C26=O22\u00a0= 1.2128\u2005(19)\u2005\u00c5 and C26\u2014O21 = 1.326\u2005(2)\u2005\u00c5, the latter involving the OH group. The planes of the cyclo\u00adpenta\u00addienyl (Cp) rings are almost parallel to the planes of the corresponding carb\u00adoxy/carboxyl\u00adate groups, with O\u2014C\u2014C\u2014C torsion angles less than 13\u00b0. The conformation of 1,1\u2032-disubstituted ferrocenes is described by the torsion angle Csubst\u2014Cpcent\u2014Cpcent\u2014Csubst, where Csubst stands for a ferrocene C atom with an additional bonding partner and Cpcent for the centre of gravity of the C atoms of the ring; this angle is hereafter referred to as \u03c6. In (1), the anion possesses an eclipsed conformation with \u03c6 = 66.0\u00b0 consists of one 3-amino\u00adpyridinium cation and one 1\u2032-carb\u00adoxy\u00adferrocene-1-carboxyl\u00adate monoanion Fig.\u00a01. In the \u00b0) Fig.\u00a02.\u2212O2C hydrogen bonds of medium\u2013strong-to-weak nature and of CO2H\u22ef\u2212O2C hydrogen bonds of strong nature by charge-supported NH\u22efe Table\u00a02.et al., 20162C-\u03b75-C5H4)Fe(\u03b75-C5H4-CO2\u2212) units from 14 crystallographically independent monoanions. Among these 14 fragments, three adopt a trans-staggered conformation, with m = 5 was added dropwise to 1,1\u2032-di\u00adacetyl\u00adferrocene under stirring at a temperature of 317\u2013320\u2005K. The solution was stirred at this temperature for 2\u2005h. Three more 25\u2005ml portions of NaOCl solution were added every 2\u2005h. The reaction mixture was filtered and acidified to a pH of 1.1 with 10% hydro\u00adchloric acid and cooled to 277\u2005K overnight. The yellow precipitate which formed was filtered off and recrystallized from ethanol to give an orange microcrystalline powder .Ferrocene-1,1\u2032-di\u00adcarb\u00adoxy\u00adlic acid was dissolved in methanol and mixed with a methano\u00adlic solution of 3-amino\u00adpyridine . The reaction mixture was filtered and subjected to slow evaporation at room temperature to give orange crystals of the title salt.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017007058/wm5388sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017007058/wm5388Isup2.hklStructure factors: contains datablock(s) I. DOI: 1444115CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E configuration with respect to the azomethine bond, and one nitrate anion.The title aroyl hydrazone Schiff base salt, consists of one mol\u00adecular cation in the keto tautomeric form, adopting an 13H12N3O2+\u00b7N O3\u2212, consists of one mol\u00adecular cation in the keto tautomeric form, adopting an E configuration with respect to the azomethine bond, and one nitrate anion. The two units are linked via an N\u2014H\u22efO hydrogen bond. The mol\u00adecule overall is non-planar, with the pyridinium and benzene rings being inclined to each other by 4.21\u2005(4)\u00b0. In the crystal, cations and anions are linked via inter\u00admolecular O\u2014H\u22efO and bifurcated N\u2014H\u22efO hydrogen bonds, forming a two-dimensional network parallel to (101). These networks are further linked by C\u2014H\u22efO hydrogen bonds, forming slabs parallel to (101). The slabs are linked by offset \u03c0\u2013\u03c0 inter\u00adactions, involving the benzene and pyridinium rings of adjacent slabs [inter\u00adcentroid distance = 3.610\u2005(2)\u2005\u00c5], forming a three-dimensional structure. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efO/O\u22efH (45.1%), H\u22efH (19.3%), H\u22efC/C\u22efH (14.5%), H\u22efN/N\u22efH (7.9%) and C\u22efC (6.0%) inter\u00adactions.The asymmetric unit of the title aroyl hydrazone Schiff base salt, C The orgE, where torsion angle N1\u2014N2\u2014C8\u2014C9 is \u2212177.58\u2005(14)\u00b0. On the other hand, torsion angles N2\u2014N1\u2014C7\u2014C5 and C8\u2014N2\u2014N1\u2014C7 are \u2212179.66\u2005(13) and \u2212178.09\u2005(15)\u00b0, respectively, and the benzene (C1\u2013C6) and pyridinium (N3/C9\u2013C13) rings are oriented at a dihedral angle of 4.21\u2005(4)\u00b0, probably due to the steric inter\u00adactions between the hydrogen atoms (Table\u00a02The configuration at the N2=C8 [1.276\u2005(2)\u2005\u00c5] bond is s Table\u00a02. Thus, tHydr\u22efOHydrz, N\u2014HPym\u22efON and bifurcated N\u2014HHydrz\u22efON hydrogen bonds ] link adjacent layers, forming slabs parallel to (101); see Fig.\u00a03Cg1 is the centroid of atoms C1\u2013C6) and pyridinium (Cg2 is the centroid of atoms N3/C9\u2013C13) rings of adjacent slabs has an inter\u00adcentroid Cg1\u22efCg2 distance of 3.610\u2005(2)\u2005\u00c5, while \u03b1 is 4.2\u2005(1)\u00b0, and the inter\u00adplanar distances are 3.263\u2005(7) and 3.366\u2005(7)\u2005\u00c5, with an offset distance of 1.303\u2005\u00c5.Hydrogen bonding and van der Waals contacts are the dominant inter\u00adactions in the crystal packing. In the crystal, O\u2014Hs Table\u00a01 link theCrystal Explorer 17.5 , the 19.3% contribution to the overall crystal packing is reflected as widely scattered points of high density due to the large hydrogen content of the mol\u00adecule. The single spike in the centre at de = di = 1.2\u2005\u00c5 in Fig.\u00a05c is due to the short inter\u00adatomic H \u22ef H contacts and C\u22efO/O\u22efC contacts in the structure with 3.4% and 1.9% contributions to the HS have nearly symmetrical distributions of points, with the scattered points of low densities.In order to visualize the inter\u00admolecular inter\u00adactions in the crystal of the title aroyl hydrazone Schiff base salt, a Hirshfeld surface (HS) analysis 2\u00b74H2O and 4-[(4-hy\u00addroxybenzo\u00adyl)hydrazonemeth\u00adyl]pyridin in ethanol by using a branched-tube method : 3526 m, \u03bd(OH); 1375 m, \u03bd(N\u2014O); 1644 s, \u03bd(C=N); 1501 s, \u03bd(N=O); 1105 s, \u03bd(NN).The title compound was prepared by the reaction of Cd(NOUiso(H) = 1.2Ueq(C). The highest residual electron density was found 2.48\u2005\u00c5 from atom H1.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018002141/su5422sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018002141/su5422Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018002141/su5422Isup3.cmlSupporting information file. DOI: 1822116CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Two new salts \u2013 2,6-di\u00adamino-4-chloro\u00adpyrimidin-1-ium 5-chloro\u00adsalicylate and bis\u00ad naphthalene-1,5-di\u00adsulfonate \u2013 have been synthesized and characterized by single-crystal X-ray diffraction. The supra\u00admolecular inter\u00adactions such as hydrogen bonding, halogen bonding, C\u2014Cl\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions are investigated for these crystal structures. 4H6ClN4+\u00b7C7H4ClO3\u2212, (I), and bis\u00ad naphthalene-1,5-di-sulfonate, 2C4H6ClN4+\u00b7C10H6O6S22\u2212, (II), have been synthesized and characterized by single-crystal X-ray diffraction. In both compounds, the N atom of the pyrimidine group in between the amino substituents is protonated and the pyrimidinium cation forms a pair of N\u2014H\u22efO hydrogen bonds with the carboxyl\u00adate/sulfonate ion, leading to a robust R22(8) motif (supra\u00admolecular heterosynthon). In compound (I), a self-complementary base pairing involving the other pyrimidinium ring nitro\u00adgen atom and one of the amino groups via a pair of N\u2014H\u22efN hydrogen bonds [R22(8) homosynthon] is also present. In compound (II), the crystallographic inversion centre coincides with the inversion centre of the naphthalene-1,5-di\u00adsulfonate ion and all the sulfonate O atoms are hydrogen-bond acceptors, generating fused-ring motifs and a quadruple DDAA array. A halogen-bond (Cl\u22efCl) inter\u00adaction is present in (I) with a distance and angle of 3.3505\u2005(12)\u2005\u00c5 and 151.37\u2005(10)\u00b0, respectively. In addition, a C\u2014Cl\u22ef\u03c0 inter\u00adaction and a \u03c0\u2013\u03c0 inter\u00adaction in (I) and a \u03c0\u2013\u03c0 inter\u00adaction in (II) further stabilize these crystal structures.The crystals of two new salts, 2,6-di\u00adamino-4-chloro\u00adpyrimidin-1-ium 5-chloro\u00adsalicylate, C The pyrimidine group offers two protonation sites (the two ring nitro\u00adgens) and the site of protonation depends on the nature of the substituents. Tautomerism of the pyrimidinium cation has also been reported recently via two N\u2014H\u22efO bonds (Table\u00a01S(6) motif (involving the carboxyl group and the phenolic \u2013OH) observed in salicylates/salicylic acid is also present hydrogen bonds forming an et al., 2014et al., 2012aet al., 2012bvia a Cl\u22efCl inter\u00adaction \u2005\u00c5; symmetry code: x, \u22121\u00a0+\u00a0y, z; Cg1 and Cg2 are the centroids of the N1/C2/N3/C4/C5/C6 and C8\u2013C13 rings, respectively] and C\u2014Cl\u22ef\u03c0 inter\u00adactions s Table\u00a01 forming et al., 1995et al., 2007et al., 2001et al., 2002et al., 2003et al., 2007et al., 2007via N\u2014H\u22efO bonds. Weak \u03c0\u2013\u03c0 stacking inter\u00adactions is also present 8) Fig.\u00a05. This tynt Fig.\u00a06.et al., 2014et al., 2017H-benzimidazole-3-ium) naphthalene-1,5-di\u00adsulfonate Uiso(H) = kUeq, where k = 1.5 for hy\u00addroxy and 1.2 for all other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018001196/zl2723sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989018001196/zl2723Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989018001196/zl2723IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989018001196/zl2723Isup4.cmlSupporting information file. DOI: 1817972, 1817971CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title Schiff base compound is considerably non-planar, with the outer phenol and pyridine rings being inclined to each other by 70.21\u2005(3)\u00b0. 19H17N3O, the configuration about the C=N bond is E. The mol\u00adecule is non-planar, with the phenolic and pyridine rings being inclined to the central benzene ring by 56.59\u2005(4) and 15.13\u2005(14)\u00b0, respectively. In the crystal, mol\u00adecules are linked by pairs of O\u2014H\u22efN hydrogen bonds, forming inversion dimers. The dimers are connected to neighbouring dimers by N\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming layers parallel to the bc plane. The layers are linked by offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.779\u2005(2)\u2005\u00c5], forming a three-dimensional supra\u00admolecular structure. Quantum chemical calculations of the mol\u00adecule are in good agreement with the solid-state structure.In the title Schiff base compound, C The configuration about the C14=N2 bond is E, with a C11\u2014N2\u2014C14\u2014C15 torsion angle of 176.40\u2005(2)\u00b0. The C7\u2014N1\u2014C8 angle is 123.43\u2005(1)\u00b0 and the C7\u2014N1\u2014H1A\u2014C8 fragment is approximately planar; the amine N1 atom exhibits a geometry what is typical for an sp2 rather than an sp3 atom. Bond angles C11\u2014N2\u2014C14 and C15\u2014N3\u2014C19 are also near 120\u00b0 , and the imine group has a torsion angle C11\u2014N2\u2014C14\u2014C15 of 176.40\u2005(2)\u00b0.The mol\u00adecular structure of the title compound is illustrated in Fig.\u00a01bc plane \u2005\u00c5; Cg is the centroid of the N3/C15\u2013C19 ring; inter\u00adplanar distance = 3.462\u2005(1)\u2005\u00c5 and slippage = 1.516\u2005\u00c5; symmetry code (iii) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01].In the crystal, pairs of O\u2014H\u22efN hydrogen bonds link the mol\u00adecules to form inversion dimers, with an f Table\u00a01. The dimf Table\u00a01 and C\u2014H\u22eff Table\u00a01, formingne Fig.\u00a03. The slaet al., 2016i.e. N-(2-pyridyl\u00admethyl\u00adene)benzene-1,4-di\u00adamine \u00b0 and the N atoms are also trans to each another. This is in contrast to the situation in the metal complexes of EXOQAK, e.g. di\u00adchloro\u00ad{N-[(pyridin-2-yl)methyl\u00adene]benz\u00adene-1,4-di\u00adamine}\u00adzinc(II) level and lowest unoccupied orbitals (LUMO) are named frontier orbitals (FMOs). The LUMO and HOMO orbital energy parameters are considerably answerable for the charge transfer, chemical reactivity and kinetic/thermodynamic stability of a mol\u00adecule 1. The DFT study of the title compound revealed that the HOMO and LUMO are localized in the plane extending from the whole phenol ring to the pyridine ring and electron distribution of the HOMO-1, HOMO, LUMO and the LUMO+1 energy levels are shown in Fig.\u00a04The title compound was prepared from an equimolar mixture of 4-amino\u00adphenyl\u00adamino\u00admethyl\u00adphenol and pyridine-2-carbaldehyde in (50\u2005ml) methanol. The yellow reaction mixture was stirred for 3\u2005h at room temperature and solvent was evaporated to 5\u2005ml. The resulting yellow solid was isolated by filtration, washed successively with a cold water and methanol mixture (10\u2005ml) and hexane (20\u2005ml). The compound was recrystallized from hot methanol, giving yellow plate-like crystals. Finally, the yellow solid was dried in a vacuum desiccator .max nm : 258 , 383 . IR : \u03bd(C=N) 1625, \u03bd(N\u2014H) 3265.Spectroscopic data: UV\u2013Vis (MeOH): \u03bb1H NMR : \u03b4 8.6 , 7.4 , 7.8 , 8.0 , 8.5 , 6.7 , 6.6 , 4.1 , 7.1 , 7.2 , 9.3 (\u2013OH), 6.5 (NH).m/z [M + H]+ calculated for C19H17N3O: 304.1444; found: 304.1455.HRMS (ESI) ca 40% of the data can be considered to be observed; hence the large value for Rint of 0.122. The N\u2014H and O\u2014H H atoms were located in difference Fourier maps. The OH H atom was freely refined, while during refinement, the N- and C-bound H atoms were included in calculated positions and treated as riding, with N\u2014H = 0.86\u2005\u00c5 and C\u2014H = 0.93\u2005\u00c5, and Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018003043/su5421sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989018003043/su5421Isup2.hklStructure factors: contains datablock(s) I. DOI: 1542988CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Two independent copper(II) cations with coordination numbers of 4 and 6 are bridged by dianionic salicylate anions into chains extending parallel [001]. O\u2014H\u22efO hydrogen-bonding inter\u00adactions involving both the coordinating and the lattice water mol\u00adecules result in the formation of a three-dimensional network. 2(C7H4O3)2(H2O)2]\u00b72H2O}n, contains two copper(II) cations in special positions (one on a twofold rotation axis and one on an inversion centre) and the the salicylate ligand in its dianionic form. By four- and six-coordinate metal coordination, chains are formed parallel to [001], which are extended by O\u2014H\u22efO hydrogen bonding into sheets extending parallel to (100). These sheets are weakly connected by O\u2014H\u22efO hydrogen bonding via the non-coordinating lattice water mol\u00adecules into a three-dimensional network.The title compound, {[Cu Cu2 is four-coordinated in a square-planar configuration with donor atoms O2 of the carboxyl\u00adate and O3 of the deprotonated hy\u00addroxy group. The two pairs of Cu\u2014O distances are 1.905\u2005(2)\u2005\u00c5 and are the shortest in the present structure (Table\u00a01i and the O4\u2014Cu1\u2014O4i angles . This allows the five atoms Cu1, O1, O4, O1i and O4i to deviate significantly from planarity. The sum of the cis angles is 382.8\u2005(3)\u00b0 and the dihedral angle between the O1\u2014Cu1\u2014O1i/O4\u2014Cu1\u2014O4i planes is 49.86\u2005(14)\u00b0. If one decides to consider Cu1 as four-coordinated, the coordination environment is consequently best described as halfway between square-planar and tetra\u00adhedral with approximate Dd2 symmetry \u00b72H2O \u2005\u00c5 from the least-squares plane. This small deviation involves the carboxyl\u00adate group with torsion angles of \u22123.4\u2005(5) \u00b0 for C7\u2014C2\u2014C1\u2014O1 and \u22122.5\u2005(6) \u00b0 for C3\u2014C2\u2014C1\u2014O2. The C\u2014OH bond length of 1.359\u2005(2)\u2005\u00c5 in CuO Table\u00a01. One of x = 0.25 and z = 0.25, which is the inter\u00adsection of two glide planes. By symmetry, there are four channels per unit cell with a volume of 59\u2005\u00c53 each, as calculated with the PLATON software TOPOS 0.55\u2005g (4\u2005mmol) salicylic acid were suspended in 8\u2005ml water. With a concentrated NaOH solution the pH value was adjusted to approximately 5. A solution of 0.5\u2005g (2\u2005mmol) copper(II) sulfate penta\u00adhydrate in 10\u2005ml water was added. Crystals appeared after a few days of standing. From the unit-cell determinations it became clear that the mixture of crystals contained at least three species: colourless salicylic acid, green CuCrystal data, data collection and structure refinement details are summarized in Table\u00a03ca 2\u00b0 rotation about hkl = I, global. DOI: 10.1107/S2056989017000883/wm5360Isup2.hklStructure factors: contains datablock(s) I. DOI: 1528159CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the 1,3-dioxane ring has an envelope conformation. In the crystal, classical O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link mol\u00adecules into a sheet structure, and a weak inter\u00admolecular C\u2014H\u22efCl inter\u00adaction extends the sheet structure into a three-dimensional network. 10H14Cl3NO5, the five-membered dioxolane ring adopts an envelope conformation with the C atom bonded to the butenoate side chain as the flap. It deviates from the mean plane of the other atoms in the ring by 0.446\u2005(6)\u2005\u00c5. In the crystal, mol\u00adecules are connected by O\u2014H\u22efO hydrogen bonds into helical chains running along the b-axis direction. The chains are linked into a sheet structure parallel to (001) by an N\u2014H\u22efO hydrogen bond. These classical hydrogen bonds enclose an R44(24) graph-set motif in the sheet structure. Furthermore, a weak inter\u00admolecular C\u2014H\u22efCl inter\u00adaction expands the sheet structures into a three-dimensional network.In the title compound, C On the basis of this concept, we have explored the utilization of cyclic ortho\u00adamides, prepared from allylic diol and triol with known conditions = 0.285\u2005(4)\u2005\u00c5 and \u03c6(2) = 296.7\u2005(8)\u00b0]. The C=C and C=O double bonds of the unsat\u00adurated ester are slightly skewed with torsion angle C13=C14\u2014C16=O18 being of 8.4\u2005(6)\u00b0. There is a weak intra\u00admolecular N6\u2014H6A\u22efCl1 inter\u00adaction present graph-set motif graph-set motif short contact of 3.076\u2005(3)\u2005\u00c5 is also observed.In the crystal, a classical O\u2014H\u22efO hydrogen bond , related to the title compound has not yet been reported.In the Cambridge Structural Database , one derivative , and one derivative . The amino H atoms were refined as adopting an sp2 configuration for WEKWOY and WAXBEE, while they were refined assuming an sp3 configuration of the N atom for LIBHIO and LAGMAK, as in the present study. Each N\u2014H bond of the amino group in LIBHIO is mostly eclipsed by the neighbouring C\u2014Cl bonds of the tri\u00adchloro\u00admethyl group, whereas those in the title compound are slightly tilted \u2005\u00c5; N6\u2014H6A\u22efCl1 = 115\u2005(3)\u00b0] in the title compound . HRMS (ESI) m/z calculated for C10H15Cl3NO5+ [M + H]+: 334.0016; found: 334.0016.The title compound was afforded from Uiso(H) = 1.5Ueq(methyl C) and 1.2Ueq(C) for other C-bound H atoms. The hy\u00addroxy H atom was placed, guided by difference-Fourier maps, with O\u2014H = 0.84\u2005\u00c5 and refined with Uiso(H) = 1.5Ueq(O). The amino H atoms were placed, guided by difference-Fourier maps, and were refined with distance restraints of N\u2014H = 0.86\u2005(2)\u2005\u00c5 and H\u22efH = 1.40\u2005(2)\u2005\u00c5, with Uiso(H) = 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017008283/su5377sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989017008283/su5377Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017008283/su5377Isup3.cmlSupporting information file. DOI: 1554119CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports6: Article number: 3745510.1038/srep37455; published online: 11222016; updated: 05042017This Article contains errors. The position of the mutation p.(R91W); (V172D) was incorrectly calculated, taking as a starting point the beginning of cDNA rather than the start codon. The correct position of the mutation is c.[271C\u2009>\u2009T]; [515T\u2009>\u2009A]. As a result of this the following changes in the Article are made:The legend of Figure 3 is incorrect,\u201c(C) c.[325C\u2009>\u2009T]; [569T\u2009>\u2009A] (RPE65)\u201dshould read:\u201c(C) c.[271C\u2009>\u2009T]; [515T\u2009>\u2009A] (RPE65)\u201d.In Table 3 for the family ID F7, the DNA mutation should read \u2018c.[271C\u2009>\u2009T]; [515T\u2009>\u2009A]\u2019.In Figure 5 for family F7 \u2018RPE65, c.325C\u2009>\u2009T; 569T\u2009>\u2009A; p.R91W; V172D\u2019 should read \u2018RPE65, c.271C\u2009>\u2009T; 515T\u2009>\u2009A; p.R91W; V172D\u2019. The correct Figure 5 appears below as In the Results section under the subheading \u2018Mutation analysis\u2019,\u201cThe second mutation was identified in family F7 and was a new compound heterozygous mutations c.[325C\u2009>\u2009T]; [569T\u2009>\u2009A] resulting in p.(R91W); (V172D) \u201d.should read:\u201cThe second mutation was identified in family F7 and was a new compound heterozygous mutations c.[271C\u2009>\u2009T]; [515T\u2009>\u2009A] resulting in p.(R91W); (V172D) \u201d."} +{"text": "In the crystal, mol\u00adecules are linked by O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds, forming chains propagating along [101].In the title boronic acid derivative, the mean plane of the \u2013B(OH) 7H6BNO2, the mean plane of the \u2013B(OH)2 group is twisted by 21.28\u2005(6)\u00b0 relative to the cyano\u00adphenyl ring mean plane. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds, forming chains propagating along the [101] direction. Offset \u03c0\u2013\u03c0 and B\u22ef\u03c0 stacking inter\u00adactions link the chains, forming a three-dimensional network. Hirshfeld surface analysis shows that van der Waals inter\u00adactions constitute a further major contribution to the inter\u00admolecular inter\u00adactions, with H\u22efH contacts accounting for 25.8% of the surface.In the title compound, C However, the \u2013B(OH)2 moiety contains two O\u2014H hydrogen-bond donors and can, thus, form two O\u2014H\u22efX hydrogen bonds and adopt different conformations . This enables the generation of hydrogen-bonding networks with increased dimensionality (one to three dimensions) in the solid state , alcohols (\u2013OH) and pyridines, which are based on mol\u00adecular recognition processes 2 mean plane is twisted by 21.28\u2005(6)\u00b0 relative to the cyano\u00adphenyl ring mean plane. This torsion disables intra\u00admolecular C\u2014H\u22efO hydrogen bonding between the oxygen atom of the exo-oriented B\u2014OH function and weakens the B\u2014C \u03c0\u2013\u03c0 bonding inter\u00adactions A; graph set B; graph set C; graph set a, Table\u00a01et al., 2017i, C2\u22efN1ii and O2\u22efN1ii separations in motifs A, B and C are 2.796\u2005(1), 3.452\u2005(2) and 2.909\u2005(2)\u2005\u00c5, respectively \u2005\u00c5, see Table\u00a01Cg\u22efCgiv = 3.8064\u2005(8) \u00c5; slippage 1.38\u2005\u00c5; symmetry code (iv) = \u22121\u00a0+\u00a0x, y, z] and \u03b72-type B\u22ef\u03c0 contacts with B\u22efC distances of 3.595\u2005(2) and 3.673\u2005(2)\u2005\u00c5 . Similar inter\u00adactions are also depicted in mol\u00adecular crystals formed between 1,4-benzene\u00addiboronic acid and aromatic amine N-oxides y Table\u00a01, and are\u2005\u00c5 Fig.\u00a03c. SimilCrystalExplorer and the shape-index (\u22121.0 to 1.0\u2005\u00c5). In the dnorm map, the vivid red spots in the Hirshfeld surface are due to short normalized O\u22efH and N\u22efH distances corresponding to O\u2014H\u22efO and O\u2014H\u22efN inter\u00adactions. The white spots represent the contacts resulting from C\u2014H\u22efN hydrogen bonding . On the shape-index surface for compound (I)2 group behaves simultaneously as a donor and an acceptor, meanwhile the \u2013C\u2261N group is an acceptor only. The occurrence of offset \u03c0\u2013\u03c0 inter\u00adactions is indicated by adjacent red and blue triangles .Hirshfeld surfaces and fingerprint plots were generated for (I)ng Fig.\u00a04a. On thes Fig.\u00a04b.et al., 2007The two-dimensional fingerprint plots qu\u00adantify the contributions of each type of non-covalent inter\u00adaction to the Hirshfeld surface in 5\u2005ml of ethanol was heated to reflux for 15\u2005min. The solution was left to evaporate slowly at room temperature, giving after one week colorless crystals suitable for single-crystal X-ray diffraction analysis.Uiso(H) = 1.2Ueq(C) and 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018003146/su5425sup1.cifCrystal structure: contains datablock(s) Global, I. DOI: 10.1107/S2056989018003146/su5425Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018003146/su5425Isup3.cmlSupporting information file. DOI: 1825335CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the indazole ring system is oriented at dihedral angles of 25.04\u2005(4) and 5.10\u2005(4)\u00b0 o the furan and benzene rings, respectively 20H16Cl2N2O2, the indazole ring system is approximately planar [maximum deviation = 0.033\u2005(1)\u2005\u00c5], its mean plane is oriented at dihedral angles of 25.04\u2005(4) and 5.10\u2005(4)\u00b0 to the furan and benzene rings, respectively. In the crystal, pairs of C\u2014Hind\u22efObo (ind = indazole and bo = benz\u00adyloxy) hydrogen bonds link the mol\u00adecules into centrosymmetric dimers with graph-set motif R22(12). Weak C\u2014H\u22ef\u03c0 inter\u00adactions is also observed. Aromatic \u03c0\u2013\u03c0 stacking between the benzene and the pyrazole rings from neighbouring mol\u00adecules [centroid\u2013centroid distance = 3.8894\u2005(7)\u2005\u00c5] further consolidates the crystal packing.In the title compound, C Its mean plane is oriented with respect to the furan and benzene rings at dihedral angles of A/B = 25.04\u2005(4) and B/C = 5.10\u2005(4)\u00b0. The dihedral angle between the furan and benzene rings is 20.21\u2005(5)\u00b0. Atom C6 is \u22120.054\u2005(1)\u2005\u00c5 from the indazole ring plane, while atom C1 is 0.038\u2005(1)\u2005\u00c5 from the furan ring plane. Atoms Cl1, Cl2 and C14 are displaced by \u22120.0430\u2005(3), 0.0233\u2005(4) and \u22120.016\u2005(1)\u2005\u00c5, respectively, to the benzene ring plane.In the mol\u00adecule of the title compound, Fig.\u00a01, the bonind\u22ef Obo (ind = indazole and bo = benz\u00adyloxy) hydrogen bonds \u2005\u00c5; symmetry code: (i) 2\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y, \u2212 z; Cg3 and Cg4 are the centroids of rings C (C15\u2013C20) and D (N1/N2/C7/C8/C13)].In the crystal, pairs of C\u2014Hs Table\u00a01, enclosirs Fig.\u00a02, which aon Fig.\u00a03. Weak C\u2014on Fig.\u00a03 occur. \u03c0H-indazol-2-yl)ethanol with NaH and 2,5-dichlorobenzyl bromide. NaH was added in small fractions to a solution of alcohol in DMF (3\u20134\u2005ml). Then, 2,5-dichlorobenzyl bromide was added portionwise. The mixture was stirred at room temperature for 3\u2005h, and the excess hydride was decomposed with a small amount of methyl alcohol. After evaporation to dryness under reduced pressure, a small amount of water was added and extracted with methyl\u00adene chloride. The organic layer was separated, dried over anhydrous sodium sulfate, and then evaporated to dryness. The crude residue was purified by chromatography on a silica-gel column using a hexa\u00adne\u2013ethyl acetate mixture (10:1) as eluent. The ether was recrystallized from 2-propanol solution to obtain colourless crystals suitable for X-ray analysis .The title compound was synthesized by the reaction of 1-(furan-2-yl)-2-(2Uiso(H) = 1.2Ueq(C).The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0210.1107/S2056989016013827/xu5891sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016013827/xu5891Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016013827/xu5891Isup3.cmlSupporting information file. DOI: 1501341CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(6) ring motif in the title compound. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds with a C(4) chain motif, and also by C\u2014H\u22ef\u03c0 inter\u00adactions. The chains are linked by \u03c0\u2013\u03c0 inter\u00adactions, forming a sheet parallel to the bc plane.There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond forming an 15H18N2O3, the coumarin ring is essentially planar, with an r.m.s. deviation of 0.012\u2005\u00c5. An intra\u00admolecular O\u2014H\u22efN hydrogen bond forms an S(6) ring motif. The piperazine ring adopts a chair conformation. In the crystal, a C\u2014H\u22efO hydrogen bond generates a C(4) chain motif running along the c axis. The chain structure is stabilized by a C\u2014H\u22ef\u03c0 inter\u00adaction. The chains are linked by \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance of 3.5745\u2005(11)\u2005\u00c5], forming a sheet structure parallel to the bc plane.In the title compound, C H-chromen-2-one) derivatives have wide applications in diverse areas such as pharmaceuticals ring motif \u2005\u00c5, \u03b8 = 1.9\u2005(2)\u00b0 and \u03c6 = 22\u2005(7)\u00b0. The C16\u2014N4\u2014C15\u2014C14 and C19\u2014N4\u2014C15\u2014C14 torsion angles are \u221278.8\u2005(2) and 158.52\u2005(16)\u00b0, respectively. The bond lengths and angles of the title compound are normal and agree with those values in other Mannich bases of 7-hy\u00addroxy\u00adcoumarin chain motif running parallel to the c axis. A C\u2014H\u22ef\u03c0 inter\u00adaction \u2005\u00c5, inter-planar distance = 3.404\u2005\u00c5 and slippage = 1.090\u2005\u00c5; symmetry code: (iii) \u2212x, \u2212y, \u2212z\u00a0+\u00a01], forming a supra\u00admolecular sheet parallel to the bc plane.In the crystal, mol\u00adecules are linked by a C\u2014H\u22efO hydrogen bond -l-alanine : [M + H]+, 275.1. Analysis calculated for C15H18N2O3: C 65.68, H 6.61, N 10.21%; found: C 65.40, H 6.45, N 10.06%.The title compound was prepared by modification of the reported procedure (Mazzei Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016017217/is5461sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989016017217/is5461Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016017217/is5461Isup3.cmlSupporting information file. DOI: 1511659CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound comprises a central pyromellitic di\u00adimide moiety with terminal decyl groups, with potential applications as an acaricide, insecticide and mematicide. 30H44N2O4 iso\u00adindole-1,3,5,7-tetra\u00adone], consists of a central pyromellitic di\u00adimide moiety with terminal decyl groups at the N-atom positions. The centre of the mol\u00adecule lies on a crystallographic inversion centre so the asymmetric unit contains one half-mol\u00adecule. The mol\u00adecule exhibits a rod-shaped conformation, like other similar compounds of this type, the distance between the ends of terminal decyl groups being 32.45\u2005\u00c5. The packing is dominated by a lamellar arrangement of the mol\u00adecules, which is reinforced by C\u2014H\u22efO hydrogen bonds and C\u2014O\u22ef\u03c0 inter\u00adactions, forming a classic herringbone structure. The mol\u00adecular structure is consistent with the theoretical calculations performed by density functional theory (DFT).The title compound, C In addition, a reversible anti to syn transition was achieved by agitating in mixed organic solvents x, \u2212y\u00a0+\u00a0z\u00a0\u2212\u00a0ii\u22efCg1 (Cg1 is the centroid of the N1/C1\u2013C4 ring) inter\u00adactions [O\u22ef\u03c0 = 3.272\u2005(1)\u2005\u00c5; symmetry code: (ii) x, \u2212y\u00a0+\u00a0z\u00a0+\u00a0bc plane : d = 8.27 , 3.74 , 1.70 , 1.32 , 0.88 A mixture of pyromellitic dianhydride and decyl amine in toluene (10\u2005ml) and dimethyl sulfoxide (6\u2005ml) was heated at 453\u2005K with stirring for 5\u2005h. Upon cooling to room temperature, an off-white crude solid was filtered and washed with water, methanol and ether. Crystals suitable for X-ray diffraction analysis were obtained by slow evaporation of a di\u00adchloro\u00admethane solution of the title compound. d(C\u2014H) = 0.95\u2005\u00c5, Uiso(H) = 1.2Ueq(C) for aromatic C\u2014H, d(C\u2014H) = 0.99\u2005\u00c5, Uiso(H) = 1.2Ueq(C) for Csp3\u2014H, d(C\u2014H) = 0.98\u2005\u00c5, Uiso = 1.5Ueq(C) for methyl group.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017006867/hb7674sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989017006867/hb7674Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017006867/hb7674Isup3.cmlSupporting information file. DOI: 1548456CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports6: Article number: 3762710.1038/srep37627; published online: 11222016; updated: 04052017This Article contains an error in the labelling of Figure 4D, where the panel labels \u2018\u03b1CD3\u2019 and \u2018\u03b1CD3\u2009+\u2009OFS\u2019 are inverted. The correct"} +{"text": "Both complexes display a bent metallocene unit, the metal centre is coordinated in a distorted tetra\u00adhedral geometry.The crystal structures of two ansa-titanocene tri\u00adfluoro\u00admethane\u00adsulfonate complexes bearing the Me2Si(C5Me4)2 ligand are reported, namely [di\u00admethylbis\u00ad(\u03b75-tetra\u00admethyl\u00adcyclo\u00adpenta\u00addien\u00adyl)silane]titanium(III) toluene monosolvate, [Ti(CF3O3S)(C20H30Si)]\u00b7C7H8, 1, and chlorido\u00ad[di\u00admethyl\u00adbis\u00ad(\u03b75-tetra\u00admethyl\u00adcyclo\u00adpenta\u00addien\u00adyl)silane](tri\u00adfluoro\u00admethane\u00adsulfonato-\u03baO)titanium(IV), [Ti(CF3O3S)(C20H30Si)Cl], 2. Both complexes display a bent metallocene unit, the metal atom being coordinated in a distorted tetra\u00adhedral geometry, with the tri\u00adfluoro\u00admethane\u00adsulfonate anion acting as a bidentate or monodentate ligand in 1 and 2, respectively. In 1, weak \u03c0\u2013\u03c0 stacking inter\u00adactions involving the toluene solvent mol\u00adecules [centroid-to-centroid distance = 3.9491\u2005(11)\u2005\u00c5] are observed.The crystal structures of two Photolysis of the latter results in a photoreduction and elimination of the OH ligand to give a TiIII tri\u00adfluoro\u00admethane\u00adsulfonate complex. Several cycles of this synthetic model scheme for water splitting can be passed Ti].Variation of the 2Si(C5Me4)2Ti(OTf)]\u00b7C7H8 (1) and [Me2Si(C5Me4)2Ti(OTf)Cl] (2), respectively. Both metal complexes exhibit distorted tetra\u00adhedral coordination geometries and show the typical bent metallocene moiety.Figs. 11 crystallizes with one mol\u00adecule of toluene in the asymmetric unit. The crystal structure of 1 confirms the bidentate binding mode of the tri\u00adfluoro\u00admethane\u00adsulfonate ligand, which is in contrast to other complexes bearing different metallocene units 2]2Nd(\u03baO-OTf) 3]2Ce , B: 2.058\u2005(2)\u2005\u00c5].However, this binding mode is known for group 4 complexes \u2005\u00c5, which is slightly shorter compared to the bis\u00ad(penta\u00admethyl\u00adcyclo\u00adpenta\u00addien\u00adyl) compound Cp*2Ti(Cl)(OTf) .Titanocene(IV) complexes with a tri\u00adfluoro\u00admethane\u00adsulfonate ligand in a \u03ba al. 1994; Taw et al. 2003; Deacon al. 2006; Kessler al. 2011 and Gode al. 2015. The cry1, weak \u03c0\u2013\u03c0 stacking inter\u00adactions were observed between two neighbouring toluene solvent mol\u00adecules along the a axis [distance between ring centroids 3.9491\u2005(11)\u2005\u00c5 and ring slippage of 1.985\u2005\u00c5].For 2Si(C5Me4)2Ti(\u03b72-Me3SiC2SiMe3) was prepared according to a published procedure 2Ti(\u03b72-Me3SiC2SiMe3) and Yb(OTf)3 were dissolved in 30\u2005ml of toluene and heated at 333\u2005K overnight, resulting in a colour change from dark yellow to green. All volatiles were removed in vacuo and the residue was again dissolved in toluene. The solution was filtered and the solvent was evaporated in vacuum to yield complex 1 as a dark-green powder. Single crystals suitable for an X-ray analysis were obtained from a saturated toluene solution at 195\u2005K.Synthesis of 2: In an experiment which aimed at the synthesis of the above TiIII tri\u00adfluoro\u00admethane\u00adsulfonate complex 1, a batch of the alkyne complex Me2Si(C5Me4)2Ti(\u03b72-Me3SiC2SiMe3) was used that contained significant amounts of the monochloride complex Me2Si(C5Me4)2TiCl, which was formed by incomplete reduction of the dichloride complex Me2Si(C5Me4)2TiCl2 during synthesis of the alkyne complex. Reaction of the monochloride complex with Yb(OTf)3 yields the ansa-titanocene(IV) chloride tri\u00adfluoro\u00admethane\u00adsulfonate complex 2. Single crystals suitable for an X-ray analysis were obtained from a saturated toluene solution by slow cooling from 353\u2005K to room temperature.Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C) for methyl H atoms. A rotating model was used for the methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2056989016018363/rz5199sup1.cifCrystal structure: contains datablock(s) 1, 2, New_Global_Publ_Block. DOI: 10.1107/S2056989016018363/rz51991sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989016018363/rz51992sup3.hklStructure factors: contains datablock(s) 2. DOI: 1517524, 1517523CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the original publication one authIncorrect author name:C\u00e9line ChaleurCorrect author name:C\u00e9line Chauleur"} +{"text": "II complex is centrosymmetric and the mol\u00adecules are linked by O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds into the three-dimensional supra\u00admolecular network.The Mn 10H11O2)2(C10H14N2O)2(H2O)2], the MnII cation is located on an inversion centre. The four O atoms form a slightly distorted square-planar arrangement around the MnII cation, and the distorted octa\u00adhedral coordination is completed by two pyridine N atoms at distances of 2.3289\u2005(15)\u2005\u00c5. The dihedral angle between the planar carboxyl\u00adate group and the adjacent benzene ring is 87.73\u2005(16)\u00b0, while the benzene and pyridine rings are oriented at a dihedral angle of 43.03\u2005(8)\u00b0. In the crystal, the water mol\u00adecules are involved in both intra\u00admolecular (to the non-coordinating carboxyl\u00adate O atom) and inter\u00admolecular (to the amide carbonyl O atom) O\u2014H\u22efO hydrogen bonds. The latter lead to the formation of layers parallel to (100). These layers are further linked via weak C\u2014H\u22efO hydrogen bonds, resulting in a three-dimensional supra\u00admolecular network. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (70.0%), H\u22efO/O\u22efH (15.5%) and H\u22efC/C\u22efH (14.0%) inter\u00adactions. One of the ethyl groups of the di\u00adethyl\u00adnicotinamide ligand is disordered over two sets of sites, with an occupancy ratio of 0.282\u2005(10):0.718\u2005(10).In the title centrosymmetric complex, [Mn(C N,N-Di\u00adethyl\u00adnicotinamide (DENA) is an important respiratory stimulant 2(DENA)2(H2O)2] and some benzoic acid derivatives as ligands, e.g. [Zn(NA)2(C7H5O3)2] \u00b7H2O , was synthesized and its crystal structure is reported on herein.The structure\u2013function\u2013coordination relationships of the aryl\u00adcarboxyl\u00adate ion in MnII cation located on an inversion centre, one 2,4,6-tri\u00admethyl\u00adbenzoate (TMB) anion and one N,N-di\u00adethyl\u00adnicotinamide (DENA) mol\u00adecule together with the one water mol\u00adecule, with all ligands coordinating to the MnII cation in a monodentate manner of the two symmetry-related TMB anions and the two symmetry-related water O atoms (O4 and O4i) at distances of 2.0999\u2005(14) and 2.2230\u2005(15)\u2005\u00c5, respectively, to form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination sphere is completed by the two pyridine N atoms (N1 and N1i) at distances of 2.3289\u2005(15)\u2005\u00c5 of the two symmetry-related DENA ligands in the axial positions \u2005\u00c5]. The Mn\u2014N bond length [2.3289\u2005(15)\u2005\u00c5] is the longest one in the MnO4N2 octa\u00adhedron. The Mn1 atom lies 0.0697\u2005(1)\u2005\u00c5 above the planar (O1/O2/C1) carboxyl\u00adate group. The O2\u2014C1\u2014O1 bond angle [125.5\u2005(2)\u00b0] seems to be significantly increased than that present in a free acid [122.2\u00b0], in which the O2\u2014C1\u2014O1 bond angle may be compared with the corresponding values of 123.5\u2005(2) and 120.4\u2005(2)\u00b0 in (II), 119.2\u2005(3) and 123.8\u2005(2)\u00b0 in (III), 123.86\u2005(13) and 118.49\u2005(14)\u00b0 in (IV), 125.11\u2005(13) and 124.80\u2005(14)\u00b0 in (V) and 126.65\u2005(14)\u00b0 in (VI), where the benzoate ions are coordinated to the metal atoms only bidentately in (V), only monodentately in (VI) and both monodentately and bidentately in (II), (III) and (IV). The O\u2014Mn\u2014O and O\u2013Mn\u2014N bond angles deviate slightly from ideal values, with same average values of 90.00\u2005(6)\u00b0.The near equalities of the C1\u2014O1 [1.254\u2005(3)\u2005\u00c5] and C1\u2014O2 [1.243\u2005(3)\u2005\u00c5] bonds in the carboxyl\u00adate groups indicate delocalized bonding arrangements, rather than localized single and double bonds. The Mn\u2014O bond lengths [2.2230\u2005(15)\u2005\u00c5] for water oxygen atoms are by A (C2\u2013C7) ring is 87.73\u2005(16)\u00b0, while the benzene A and pyridine B (N1/C11\u2013C15) rings are oriented at a dihedral angle of A/B = 43.03\u2005(8)\u00b0.The dihedral angle between the planar carboxyl\u00adate group (O1/O2/C1) and the adjacent benzene w\u22efOc hydrogen bonds (Table\u00a01S(6) ring motif hydrogen bonds and C\u2014HDENA\u22efODENA hdyrogen bonds analysis in H2O (100\u2005ml) and N,N-di\u00adethyl\u00adnicotinamide in H2O (10\u2005ml) with sodium 2,4,6-tri\u00admethyl\u00adbenzoate in H2O (150\u2005ml). The mixture was filtered and set aside to crystallize at ambient temperature for three weeks, giving colourless single crystals.The title compound was prepared by the reaction of MnSOUiso(H) = k \u00d7 Ueq(C), where k = 1.5 for methyl H atoms and k = 1.2 for other H atoms. The disordered ethyl group was refined over two sets of sites with distance restraints and SIMU and DELU restraints I, global. DOI: 10.1107/S2056989018003377/xu5920Isup2.hklStructure factors: contains datablock(s) I. DOI: 1826038CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The current redetermination confirms the previous structure report, but with considerably higher precision and accuracy. 3)2(C5H7O2)2] or SnMe2(acac)2, from CCD data recorded at 100\u2005K basically confirms the previous study based on integrated film data recorded at room temperature [Miller & Schlemper > 6.7694\u2005(4)\u2005\u00c5]. The mol\u00adecule belongs to point group iC with the SnIV atom on a centre of inversion. The SnIV atom shows a slightly distorted octa\u00adhedral coordination sphere with the methyl groups in trans positions and a Sn\u2014C bond length of 2.115\u2005(2)\u2005\u00c5 which may serve as a standard value for an Sn\u2014CH3 bond of an octa\u00adhedrally coordinated SnIV atom. The Sn\u2014O bonds involving the two carbonyl groups of the acetyl\u00adacetonate ligand are of equal length [2.180\u2005(1) and 2.183\u2005(1)\u2005\u00c5], as are the C=O [1.273\u2005(1) and 1.274\u2005(1)\u2005\u00c5] and C\u2014C bond lengths [1.393\u2005(2) and 1.400\u2005(2)\u2005\u00c5]. The acetyl\u00adacetonate ligand deviates considerably from planarity, with a dihedral angle of 5.57\u2005(9)\u00b0 between the least-squares planes of the two acetone moieties. The four O atoms of the two symmetry-related acetyl\u00adacetonate ligands are arranged in a nearly quadratic rectangle. Weak C\u2014H\u22efO inter\u00adactions consolidate the crystal packing.The redetermination of the title compound, [Sn(CHmper 1972. Inorg. Moreover, the title compound is an excellent candidate for the determination of the Sn\u2014CMe bond length as another reference in case the SnIV atom is in a well-defined octa\u00adhedral coordination. The precise measurement of this Sn\u2014C distance therefore should supplement the observations of Britton and the constitution of the asymmetric unit comprising half a formula unit with the Sn atom at a crystallographic centre of inversion [Wyckhoff symbol: b]. As we performed the X-ray measurement at 100\u2005K, the unit-cell volume is somewhat smaller in comparison with the original room-temperature data which is mainly caused by a considerable change of the a axis from 7.12\u2005(1) to 6.7694\u2005(4)\u2005\u00c5 while changes of all other lattice parameters show a normal temperature-dependent shrinkage.The redetermination of the crystal structure of the title compound confirms the former results obtained by Miller & Schlemper 1973 with restrans position \u00b0. The four O atoms of the two symmetry-related acetyl\u00adacetonate ligands around the Sn atom form a planar rectangle with similar edge lengths [O1\u22efO2 = 2.975\u2005(1)/O1\u22efO1 = 3.191\u2005(1)\u2005\u00c5], and almost right angles [89.9\u2005(1)\u00b0 at O1 and 90.1\u2005(1)\u00b0 at O2]. This plane is nearly perpendicular to the axis through the two methyl groups [deviation: 0.44\u2005(2)\u00b0] but constitutes a dihedral angle of 10.2\u2005(1)\u00b0 with the least-squares plane through the two carbonyl groups of the acetyl\u00adacetonate ligand x, y, 1\u00a0+\u00a0z; (ii) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y,1\u00a0\u2212\u00a0z; (iii) x, y, z; (iv) x, \u2212y, z] 2\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z] for each oxygen atom. In summary, the inter\u00admolecular contacts result in a columnar arrangement of the mol\u00adecules parallel to the a axis . The crystals are stable in air.The synthesis of the title compound by refluxing a suspension of di\u00admethyl\u00adtin oxide, Mebias 1965. Single 1H NMR , nJ [Hz]): \u03b4(CH3\u2014Sn) = 0.58, 2J(1H\u2014119/117Sn) = 100.6/97.1; \u03b4(CH3)acac = 1.96; \u03b4(CH)acac = 5.31 ], nJ [Hz]): \u03b4(CH3\u2014Sn) = 7.75, 1J(13C\u2014119/117Sn = 973.7/930.4), \u03b4(CH3)acac = 27.94, \u03b4(CH)acac = 100.09, \u03b4(C=O)acac = 190.75 ]: 3010 w, 2920 w,1562 s, 1512 s, 1436 m, 1361 s,bd, 1256 m, 1203 m, 1015 m, 925 m, 803 m, 781 m, 655 m, 572 m, 552 m ]: 3092 w, 2999 w, 2920 s, 2708 w, 1574 w, 1427 w, 1366 m, 1263 m, 1206 m, 1194 m, 1021 w, 927 m, 668 m, 567 m, 512 s, 415 m, 220 m, 130 m 94 m, 68 m , and 0.95\u2005\u00c5 (\u2013CH\u2013) and with Uiso(H) = 1.2 and 1.5Ueq(C), respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017003206/wm5370sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989017003206/wm5370Isup2.hklStructure factors: contains datablock(s) I. DOI: 1534819CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Ti and Al(Ga), and in a \u03bc2 manner.The mol\u00adecular structures of two isotypic titanium(III) complexes bearing an tri\u00admethyl\u00adaluminium or -gallium motif are reported. In both compounds, two methyl groups coordinate to the metal atoms, EMe3 unit with two \u03bc2-coordinating methyl groups, namely [\u03bc-1(\u03b75)-(adamantan-1-yl-2\u03baC1)cycylo\u00adpenta\u00addien\u00adyl]di-\u03bc2-methyl-methyl-2\u03baC-[1(\u03b75)-penta\u00admethyl\u00adcyclo\u00adpenta\u00addien\u00adyl]aluminiumtitanium(III), [AlTi(CH3)3(C10H15)(C15H18)], and [\u03bc-1(\u03b75)-(adamantan-1-yl-2\u03baC1)cycylo\u00adpenta\u00addien\u00adyl]di-\u03bc2-methyl-methyl-2\u03baC-[1(\u03b75)-penta\u00admethyl\u00adcyclo\u00adpenta\u00addien\u00adyl]galliumtitanium(III), [GaTi(CH3)3(C10H15)(C15H18)], are reported. Reacting a dinuclear nitro\u00adgen-bridged low-valent titanium(III) complex with the Lewis acids AlMe3 or GaMe3 results in the loss of mol\u00adecular di\u00adnitro\u00adgen and the formation of two monomeric titanocene(III) fragments bearing two \u03bc2-bridging methyl groups. Single crystal X-ray diffraction reveals the formation of a new E\u2014C bond involving the penta\u00adfulvene ligand while the bridging and terminal methyl groups remain intact.The isotypic crystal structures of two titanocene complexes containing an The crystal packing Fig.\u00a03 appears et al., 20053 and GaMe3 solutions were purchased from Sigma Aldrich and used as received. Solvents were dried according to standard procedures over Na/K alloy with benzo\u00adphenone as indicator and distilled under a nitro\u00adgen atmosphere.All reactions were carried out under a dry nitro\u00adgen atmosphere using Schlenk techniques or in a glove box. The starting titanium complex was prepared according to a published procedure (Scherer Synthesis of 1:\u03b75-penta\u00admethyl\u00adcyclo\u00adpenta\u00addien\u00adyl)(\u03b75:\u03b71-adamantylidene\u00adpenta\u00adfulvene)titanium]-\u03bc2,\u03b71,\u03b71-di\u00adnitro\u00adgen was dissolved in toluene and AlMe3 was added. The colour of the solution changed from blue to green, after 48\u2005h the volume had reduced to 5\u2005ml and another 5\u2005ml of n-hexane were added. Crystals suitable for X-ray diffraction separated after 48\u2005h directly from the mother liquor.Bis[(Synthesis of 2:\u03b75-penta\u00admethyl\u00adcyclo\u00adpenta\u00addien\u00adyl)(\u03b75:\u03b71-adamantylidene\u00adpenta\u00adfulvene)titanium]-\u03bc2,\u03b71,\u03b71-di\u00adnitro\u00adgen was dissolved in toluene and GaMe3 was added. The former blue solution turned brown and was stored at 233\u2005K. After 10 days, brown\u2013green crystals suitable for X-ray diffraction separated from the mother liquor.Bis[(Uiso(H) = 1.2Ueq(C); H atoms of all methyl groups were refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2056989017004856/wm5378sup1.cifCrystal structure: contains datablock(s) global, 1, 2. DOI: 10.1107/S2056989017004856/wm53781sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989017004856/wm53782sup3.hklStructure factors: contains datablock(s) 2. DOI: 1540662, 1540661CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A mol\u00adecular docking calculation of the title compound with the neuraminidase enzyme was carried out.In the title chalcone-thio\u00adphene derivative, the dihedral angle between the aromatic and the thio\u00adphene rings is 11.4\u2005(2)\u00b0. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO and C\u2014H\u22efS weak inter\u00adactions along [100], forming rings of 15H15NOS, whose mol\u00adecular structure matches the asymmetric unit. The mol\u00adecule is not planar, the dihedral angle between the aromatic and the thio\u00adphene rings being 11.4\u2005(2)\u00b0. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO and weak C\u2014H\u22efS inter\u00adactions along [100], forming 22R(8) rings, and by weak C\u2014H\u22efO inter\u00adactions along [010], forming chains with a C(6) graph-set motif. In addition, mol\u00adecules are connected into centrosymmetric dimers by weak C\u2014H\u22ef\u03c0 inter\u00adactions, as indicated by the Hirshfeld surface analysis. The most important contributions for the crystal structure are the H\u22efH (46.50%) and H\u22efC (23.40%) inter\u00adactions. The crystal packing resembles a herringbone arrangement when viewed along [100]. A mol\u00adecular docking calculation of the title compound with the neuraminidase enzyme was carried out. The enzyme shows (ASN263)N\u2014H\u22efO, (PRO245)C\u2014H\u22efCg(thio\u00adphene ring) and (AGR287)C\u2014H\u22efN inter\u00admolecular inter\u00adactions with the title compound. The crystal structure was refined as a two-component twin with a fractional contribution to the minor domain of 0.0181\u2005(8).The equimolar reaction between 4-(di\u00admethyl\u00adamino)\u00adbenzaldehyde and 2-acetyl\u00adthio\u00adphene in basic ethano\u00adlic solution yields the title compound, C The 22R(8) rings are the subunits of the periodic arrangement along [100] and one very weak H7\u22efH2i contact is also observed [H\u22efH = 2.26\u2005\u00c5]. The mol\u00adecular units are also linked by very weak C15\u2014H15\u22efO1ii links into chains along [010] with a C(6) graph-set motif and di (x axis) values are the closest external and inter\u00adnal distances from given points on the Hirshfeld surface contacts N\u2014H\u22efO1 (d = 1.796), (PRO245)C\u2014H\u22efCg(thio\u00adphene ring) (d = 2.829) and (AGR287)C\u2014H\u22efN1 (d = 2.620) contact is observed in the structure inter\u00adpretation, by the centrosymmetric dimeric arrangement of the mol\u00adecules Fig.\u00a08. More des Figs.\u00a03 and 9 \u25b8,sp2-hybridized C atoms of the main fragment, and the weak inter\u00admolecular inter\u00adactions, e.g. H\u22efH, H\u22efC or \u03c0\u2013\u03c0 contacts. One example for comparison with the title compound is the crystal structure of the compound 3-(4-methyl\u00adphen\u00adyl)-1-(3-thien\u00adyl)-2-propen-1-one and for the above-mentioned 3-thienyl derivative, along [001] .Chalcone-thio\u00adphene derivatives have some mol\u00adecular structural features in common, namely the nearly planar geometry, as a result of the 1] Fig.\u00a09b.et al., 2016All starting materials are commercially available and were used without further purification. The synthesis of the title compound was adapted from a previously reported procedure = 1.2Ueq(C) or 1.5Ueq(C) for methyl H atoms. A rotating model was used for the methyl groups. The crystal was refined as a two-component twin {twin law: two-axis (001) [105], BASF = 0.0181\u2005(8)}.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017003437/rz5205sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017003437/rz5205Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017003437/rz5205sup3.pdfSUPPORTING INFORMATION FOR THE EVALUATION OF THE CRYSTAL STRUCTURE OF (2E)-3-[4-(DIMETHYLAMINO)PHENYL]-1-(THIOPHEN-2-YL)PROP-2-EN-1-ONE AND THE NEURAMINIDASE ENZYME. DOI: Click here for additional data file.10.1107/S2056989017003437/rz5205Isup4.cmlSupporting information file. DOI: 1535563CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure of the title compound, a di\u00adspiro\u00ad-2,5\u2032\u2032-dione, C\u2014H\u22efO hydrogen bonding predominates, linking mol\u00adecules to form chains propagating along [100]. 32H26ClN3O2, the cyclo\u00adhexa\u00adnone ring of the iso\u00adquinoline unit has a distorted envelope conformation, with the methyl\u00adene C atom adjacent to the spiro C atom as the flap. The central 1-methyl\u00adpyrrolidine ring has an envelope conformation with the N atom as the flap. The mean planes of the indolin-2-one ring system, the chloro\u00adbenzene ring and the iso\u00adquinoline ring system are inclined to the mean plane of the central 1-methyl\u00adpyrrolidine ring by 87.95\u2005(11), 71.01\u2005(12) and 88.81\u2005(10)\u00b0, respectively. There are two short C\u2014H\u22efO intra\u00admolecular contacts present. In the crystal, mol\u00adecules are linked via C\u2014H\u22ef O hydrogen bonds, forming chains along the a-axis direction. The NH H atom is involved in a weak N\u2014H\u22efO hydrogen bond with the same carbonyl O atom. There are no further significant inter\u00admolecular contacts present. The largest contribution to the overall Hirshfeld surface of 52.3% is due to H\u2014H contacts.In the title di\u00adspiro compound, C The pyridine ring (N3/C32\u2013C36) has a shallow twist-boat conformation . Their mean planes are inclined to each other by 14.06\u2005(10)\u00b0, and the phenyl ring (C51\u2013C56) is inclined to the pyridine ring mean plane by 22.35\u2005(12)\u00b0.The mol\u00adecular structure of the title mol\u00adecule is shown in Fig.\u00a01Q = 0.094\u2005(2)\u2005\u00c5, \u03b8 = 92.3\u2005(13)\u00b0, \u03c6 = 84.5\u2005(13)\u00b0]. The mean planes of the indolin-2-one ring system, the chloro\u00adbenzene (C41\u2013C46) ring and the iso\u00adquinoline (N3/C3/C31\u2013C38) ring system are inclined to the mean plane of the central 1-methyl\u00adpyrrolidine (N1/C2\u2013C5) ring by 87.95\u2005(11), 71.01\u2005(12) and 88.81\u2005(10)\u00b0, respectively. The sum of the bond angles around atoms N1 and N2 are 333.6 and 358.6\u00b0, respectively, indicating a pyramidal geometry and sp3 hybridization.In the indolin-2-one ring system (N2/C2/C21\u2013C27), the benzene (C21\u2013C26) and pyrrolidine (N2/C2/C21/C26/C27) rings make a dihedral angle of 2.45\u2005(12)\u00b0, while the keto atom O1 deviates from the attached pyrrolidine ring by 0.043\u2005(1)\u2005\u00c5. The 1-methyl\u00adpyrrole ring (N1/C2\u2013C5) has an envelope conformation with atom N1 as the flap -1,2\u2032\u2032(1\u2032\u2032H)-dione methanol solvate -dione chloro\u00adform solvate . The relative contributions of the other different inter\u00admolecular inter\u00adactions to the Hirshfeld surface in descending order are: C\u22efH/H\u22efC (23.3%), O\u22efH/H\u22efO (8.5%), Cl\u22efH/H\u22efCl (8.4%), N\u22efH/H\u22efN (4.1%) and there is only a very small contribution from other contacts, i.e. 3.1%, in the structure. This illustrates that the N\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions contribute significantly to the crystal packing of the title compound.The overall two-dimensional fingerprint plot is illustrated in Fig.\u00a05E)-6-(2-chloro\u00adbenzyl\u00adidene)-2-phenyl-7,8-di\u00adhydro\u00adquinolin-5(6H)-one (L) in 94% yield (m.p. 323\u2013324\u2005K). A mixture of isatin (1.1\u2005mmol) and sarcosine (1.1\u2005mmol) was taken in 10\u2005ml of aceto\u00adnitrile in a 50\u2005ml round-bottom flask and heated to reflux for 2\u2005h. Then 1\u2005mmol of L was added to the above reaction mixture and reflux was continued for a further 14\u2005h. After completion of the reaction, as evident from TLC, the solvent was removed under reduced pressure and the residue washed with ice-cold water (50\u2005ml). The crude product was purified by column chromatography using a 90:10 (v/v) petroleum ether\u2013ethyl acetate mixture to obtain the pure product . Colourless block-like crystals were obtained by slow evaporation of a solution in ethyl acetate.An equimolar mixture of 2-phenyl-5,6,7,8-tetra\u00adhydro-5-quinolinone and 2-chloro\u00adbenzaldehyde was dissolved in 10\u2005ml of ethanol followed by the addition of 0.5 equiv. of potassium hydroxide. The mixture was stirred for 1\u2005h at ambient temperature and the precipitate formed was filtered and dried to obtain pure (Uiso = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018005455/su5434sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018005455/su5434Isup2.hklStructure factors: contains datablock(s) I. DOI: 1835595CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "For example, conventional \u03b1\u03b2 T cells develop from DP cells after partial-agonist T cell receptor (TCR) interactions with self-peptide/MHC, whereas unconventional \u03b1\u03b2 T cells, such as TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) intraepithelial lymphocytes (IELs), require full-agonist TCR interactions. Despite this, DP cells appear homogeneous, and it remains unclear how distinct TCR signalling instructs distinct developmental outcomes. Moreover, whether TCR signals at earlier stages of development, for example in CD4(\u2212)CD8(\u2212) double negative (DN) cells, impact on later fate decisions is presently unknown. Here, we assess four strains of mice that display altered TCR signal strength in DN cells, which correlates with altered generation of unconventional TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs. FVB/n mice and mice with altered preTCR\u03b1 (pT\u03b1) expression, both displayed weaker TCR signalling in DN cells, an inefficient DN-to-DP transition, and reduced contribution of TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs to gut epithelium. Conversely, TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL development was favoured in mice with increased TCR signal strength in DN cells. Collectively, these data suggest TCR signal strength in DN cells directly impacts on subsequent DP cell differentiation, fundamentally altering the potential of thymocyte progenitors to adopt conventional versus unconventional T cell fates.CD4 Gut IELs display anti-microbial and anti-inflammatory properties and are central to the control of intestinal epithelial homeostasis2. A large proportion of gut IELs express TCR\u03b1\u03b2, and can be further characterized by expression of CD8\u03b1\u03b2 or CD8\u03b1\u03b1 dimers3. TCR\u03b1\u03b2(+)CD8\u03b1\u03b2(+) IELs are termed \u201cconventional\u201d, closely sharing gene expression signatures with CD8\u03b1\u03b2(+) T cells from secondary lymphoid organs5. By contrast, \u201cunconventional\u201d TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs do not require priming in lymphoid structures, appear restricted to the gut epithelium, and display gene expression signatures more similar to \u03b3\u03b4 T cells5.T cell receptor-expressing intraepithelial lymphocytes (IELs) are epithelial-resident T cells found at numerous body locations(+)CD8\u03b1\u03b1(+) IELs develop extra-thymically in gut lymphoid structures known as cryptopatches6. However, TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs are severely reduced in athymic nude mice, and fate-mapping experiments suggested they traverse the CD4(+)CD8(+) double positive (DP) stage in the thymus7. Further work implicated agonist self-peptide-mediated selection through TCR\u03b1\u03b2 at the DP stage8, and identified pre- and post-selection progenitor subsets9. Nonetheless, it remains unclear how \u201cstrong\u201d TCR-agonist signals in DP cells instruct positive selection of TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs instead of driving negative selection. Indeed, unconventional TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs were recently found to express TCRs that had been \u201crecycled\u201d from strong negatively selecting signals10.Initial studies suggested that unconventional TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL development, the DP stage is not the first in which TCR signalling occurs. DP cells arise from CD4(\u2212)CD8(\u2212) double negative (DN) cells in a process known as \u201c\u03b2-selection\u201d that is mediated by signalling through the preTCR (rearranged TCR\u03b2 paired with invariant pT\u03b1)11. PreTCR signalling is generally considered weak, due to very low surface preTCR expression12. By contrast, stronger signalling in DN cells, for example by TCR\u03b3\u03b4, is less efficient at generating DP cells; instead driving cells to a \u03b3\u03b4 T cell fate14.Although TCR\u03b1\u03b2 signalling at the DP stage appears critical for TCR\u03b1\u03b215. Although TCR signal strength in DN cells clearly affects the efficient induction of these processes, it is presently unclear whether it also affects the future fate of the DP cells that are generated. Here, we begin to investigate this idea in the context of TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL development. We show that FVB/n wild type (WT) mice have a much reduced TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL compartment when compared with WT C57BL/6 animals, that correlates with weaker preTCR signalling at the \u03b2-selection checkpoint. Indeed, by reducing preTCR signal strength in pT\u03b1-transgenic animals we re-capitulate this relative absence of gut TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs. By contrast, in two mouse models in which TCR signal strength is greater in DN cells by forced expression of TCR\u03b1\u03b2, increased generation of TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs was observed. Thus, these data provide evidence that TCR signal strength at the DN-to-DP transition directly influences the efficiency of TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL development.Successful transition through the \u03b2-selection checkpoint results in cell survival, extensive proliferation, and significant differentiation, events that may be mechanistically linked(+)CD8\u03b1\u03b1(+) IELs in the small intestine CD8\u03b1\u03b2(+) IELs was increased in FVB/n mice, and TCR\u03b3\u03b4(+) IELs were comparable between the two strains CD8\u03b1\u03b1(+) IELs are thought to develop in the thymus from cells that are CD4(lo)CD8(lo)TCR\u03b4(\u2212)TCR\u03b2(+)CD5(hi)CD69(+)PD-1(+)CD122(+) CD8(+) double positive (DP) cells16, that in turn are generated when the preTCR (rearranged TCR\u03b2 paired with invariant pre-T\u03b1) drives CD44(\u2212)CD25(+) DN3 cells to CD44(\u2212)CD25(\u2212) DN4 cells (and then on to the DP stage) in a process called \u03b2-selection11. C57BL/6 mice had an expected DN3-to-DN4 ratio of ~1.5, consistent with a normal transition through the \u03b2-selection checkpoint . Moreover, DN3 cells from FVB/n mice had significantly elevated surface expression of CD25, a signatory feature of inefficient progression through the DN-to-DP transition11 CD8\u03b1\u03b1(+) IEL compartment compared with C57BL/6 animals, also display inefficient developmental progression through thymic stages that lead to generation of unconventional IEL progenitors.We had noted that FVB/n wild type (WT) mice, in comparison with C57BL/6 WT mice, had significantly reduced unconventional TCR\u03b1\u03b2ine Fig.\u00a0 . By conins Fig.\u00a0. Unconvells Fig.\u00a0. IEL proint Fig.\u00a0. Howevern11 Fig.\u00a0. Thus, F(+)CD8\u03b1\u03b1(+) IEL development, we first assessed components of the preTCR. Intracellular TCR\u03b2 levels in DN3 cells appeared comparable between FVB/n and C57BL/6 animals , than C57BL/6 mice of CD71(+)i.c.TCR\u03b2(+) proliferating DN3 cells18 CD8\u03b1\u03b1(+) IELs in FVB/n mice.To investigate the inefficient DN3-to-DN4 transition in FVB/n mice that appeared to correlate with reduced unconventional TCR\u03b1\u03b2als Fig.\u00a0. For pT\u03b1ice Fig.\u00a0. It was s18 Fig.\u00a0, that exs18 Fig.\u00a0, a recog(+)CD8\u03b1\u03b1(+) IELs are linked, we generated mice in which only a weakly signalling pT\u03b1a-containing preTCR could be formed. pT\u03b1-intron-1 was removed from the full pT\u03b1 genomic locus (to prevent splicing to pT\u03b1b), in a 50\u2009kb bacterial artificial chromosome (BAC), that was then used to generate pT\u03b1a-only BAC transgenic mice on a pT\u03b1\u2212/\u2212 (C57BL/6) background DN3 cells displayed markedly less CD5, indicative of weaker TCR signalling CD8\u03b1\u03b1(+) IELs in the small intestine CD8\u03b1\u03b2(+) IELs and TCR\u03b3\u03b4(+) IELs were present in normal proportions CD8\u03b1\u03b1(+) IELs.To test the idea that weaker signalling at the \u03b2-selection checkpoint and reduced generation of unconventional TCR\u03b1\u03b2ls, Fig.\u00a0. Consistls, Fig.\u00a0, with a als Fig.\u00a0. Moreove WT Fig.\u00a0, while ning Fig.\u00a0. Importaine Fig.\u00a0 . By con(+)CD8\u03b1\u03b1(+) IELs. To test this idea further, we sought to increase TCR signalling in DN cells to ascertain if this would conversely increase unconventional TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL development. TCR\u03b1\u03b2 transgenic (tg) mice aberrantly express TCR\u03b1\u03b2 complexes that signal strongly at the \u03b2-selection checkpoint. Indeed, as well as driving DN cells to the DP stage, this strong TCR signalling is also thought to divert some DN progenitors to the \u03b3\u03b4 lineage22. OT-II mice (C57BL/6 background), express an ovalbumin-specific MHC-II-restricted V\u03b12(+)V\u03b25(+) transgenic TCR\u03b1\u03b223. V\u03b12(+) (i.e. largely tg-TCR(+)) DN3 cells from OT-II mice express much higher levels of CD5 than V\u03b12(\u2212) DN3 cells, indicative of stronger TCR signalling DN cells from OT-II mice traverse the \u03b2-selection checkpoint efficiently CD8\u03b1\u03b1(+) IELs was observed, notably inverting the TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL-to-TCR\u03b1\u03b2(+)CD8\u03b1\u03b2(+) IEL ratio rearrange early TCR\u03b1 (and TCR\u03b2). Compared with pT\u03b1\u2212/\u2212 mice, TCR\u03b4\u2212/\u2212. pT\u03b1\u2212/\u2212 mice had an increased population of DN4 cells, the majority of which robustly expressed TCR\u03b2 CD8\u03b1\u03b1(+) IELs was observed, again inverting the TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL-to-TCR\u03b1\u03b2(+)CD8\u03b1\u03b2(+) IEL ratio CD8\u03b1\u03b1(+) IEL development.TCR-tg mice express a single TCR\u03b1/TCR\u03b2 combination that may have unusual signalling characteristics. Thus, to extend these studies, we took advantage of a little-known feature of TCR\u03b4CR\u03b2 Fig.\u00a0. Unlike als Fig.\u00a0. Moreoveing Fig.\u00a0. Thus, \u03b2tio Fig.\u00a0. Collect(+)CD8\u03b1\u03b1(+) intraepithelial lymphocytes have long been debated. Initially, the recognition that TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs, but not TCR\u03b1\u03b2(+)CD8\u03b1\u03b2(+) IELs, were present in athymic mice led to suggestion of their extra-thymic generation in gut lymphoid structures called cryptopatches (CPs)6. Indeed, CPs were shown to contain CD25(+)IL-7R\u03b1(+)c-kit(+) progenitors that could reconstitute the T cell compartments of irradiated T cell-deficient animals26. Nonetheless, that athymic mice have only 5\u201310% of the total TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs of euthymic mice conversely argued for a predominately thymic origin. The thymic generation of TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs was further supported by fate mapping experiments in which a GFP marker was activated by ROR\u03b3t-promoter-driven Cre protein7. ROR\u03b3t is expressed in thymic DP cells but not in their DN precursors. As all \u03b3\u03b4 T cells develop from DN cells, TCR\u03b3\u03b4(+) IELs were GFP(\u2212). By contrast, TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs were GFP(+), suggesting their development through a DP stage7.The developmental origins of TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs use \u201cforbidden\u201d TCR\u03b2 chains that were purged from the conventional TCR\u03b1\u03b2(+) T cell pool by superantigen-driven negative selection of DP cells in the thymus27. Nonetheless, despite evidence of \u201cTCR-agonist-selection\u201d28, identifying the thymic stages through which TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL progenitors progress has proven more problematic. Cheroutre and colleagues described a minor subset of CD8\u03b1\u03b1(+) DP cells that appeared to develop as TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs if exposed to TCR-agonist signals9. Somewhat surprisingly, these \u201ctriple positive\u201d (TP) cells then shut off CD4, CD8\u03b1 and CD8\u03b2 to become TCR\u03b1\u03b2(+)CD5(+) DN cells, before re-expressing CD8\u03b1\u03b1(+) in the gut epithelium possibly under the influence of TGF\u03b229. By contrast, the stages that precede the TP stage are less clear. As too are the reasons why TP cells survive TCR-agonist signals, rather than dying as DP cells do during TCR-agonist-driven negative selection. It is conceivable that TP cells have already entered an unconventional IEL developmental pathway, and are thus cell-intrinsically distinct from DP cells that allow them to survive agonist signals. In this regard, RhoH30, and TGF\u03b2 signalling29, have been implicated in differential thymic development of TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs versus conventional T cell subsets. Nonetheless, how unconventional IEL progenitors could adopt such a distinct gene expression profile by the TP stage, and indeed how TP cells are even generated, had not been previously elucidated.It was also recognized early that TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs; stronger TCR signalling in DN cells, as judged by CD5 surface levels and the efficiency of the DN-to-DP transition, appearing to favour TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL generation. As TP cells, like DP cells, are presumably derived from DN precursors as a result of (pre)TCR signalling, we speculate that stronger signalling at the DN stage favours TP generation. Unfortunately, we were unable to formally test this idea, as CD8\u03b1\u03b1-specific antibodies are not available and TL-tetramer staining did not work in our hands9. Nonetheless in this regard, peripheral CD4(+) cells do upregulate CD8\u03b1\u03b1 when activated through TCR\u03b1\u03b2 (by cross-linking antibodies) in the presence of TGF\u03b229, the absence of which has been correlated with reduced generation of TCR\u03b1\u03b2(+)CD5(+) IEL precursors29.In this study, we have shown that modulation of TCR signal strength at the thymic DN stage fundamentally impacts on the development of TCR\u03b1\u03b2(+)) DP cells conflicts with the notion that such signals are required for commitment to the \u03b3\u03b4 T cell lineage14. Indeed, premature expression of TCR\u03b1 in DN cells of TCR\u03b1\u03b2 transgenic mice was shown to promote differentiation of CD4(\u2212)CD8(\u2212)TCR\u03b1\u03b2(+) cells with \u03b3\u03b4 T cell-like properties22. Nonetheless, many of these studies also reported increased numbers of TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IELs, and made obvious but perhaps incorrect links between these DN TCR\u03b1\u03b2(+) cells and IEL development31. However, these \u201c\u03b3\u03b4-wannabie\u201d cells do not traverse the DP stage (unlike bone fide IEL progenitors), and do not rearrange their endogenous TCR\u03b1 chains20; something obviously required for TCR\u03b1\u03b2(+) IEL development. By way of explanation, our data now suggest that in TCR\u03b1\u03b2(+) transgenic mice at least some DN cells will transition to the DP stage as a result of stronger transgenic-TCR signalling. This could boost the TP cell to conventional DP cell ratio, which in turn would favour increased TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL development.At first glance, invoking strong TCR signals in DN cells to drive generation of CD8\u03b1\u03b1(+) IELs.Finally, what would normally provide the stronger TCR signals in DN cells to drive TP generation in the absence of prematurely expressed transgenic TCR\u03b1\u03b2? TCR\u03b1 is reported to rearrange in a small fraction of WT DN cells(+)CD8\u03b1\u03b1(+) IELs. Stronger signalling favours TCR\u03b1\u03b2(+)CD8\u03b1\u03b1(+) IEL development. By contrast, weaker signalling favours a greater contribution from conventional TCR\u03b1\u03b2(+)CD8\u03b1\u03b2(+) IELs that are likely differentiated from na\u00efve conventional CD8\u03b1\u03b2(+) T cells in the gut-associated lymphoid tissue.In sum, our findings shed new light on the early thymic stages of unconventional IEL development. We demonstrate that TCR signal strength in DN cells at the \u03b2-selection checkpoint significantly influences the development of unconventional TCR\u03b1\u03b2All experimental protocols were performed in, and approved by, The Blizard Institute, Bart\u2019s and The London School of Medicine, Queen Mary University of London.C57BL/6 and FVB/n mice were purchased from Charles River Laboratories. All mice were 6\u201312 weeks old. Mice were bred and maintained in specific pathogen-free animal facilities at Queen Mary University of London. All experiments were performed in compliance with relevant laws and institutional guidelines and were approved by a local ethics committee.http://recombineering.ncifcrf.gov)33. In brief, a pT\u03b1 locus-containing BAC (BMQ452P20) was purchased from www.ensemble.org. A GalK cassette was first amplified from a GalK plasmid using primers that contained 50\u2009bp homology with sections of pT\u03b1 intron-1;BAC-transgenic mice were generated by the recombineering technique in fluorescence-activated cell sorting (FACS) buffer containing; 2% heat-inactivated fetal calf serum (FCS) (Life technologies); and 5\u2009mM ethylenediaminetetraacetic acid (EDTA) (Life technologies)). For spleen, red blood cells were removed by gradient centrifiguation at 1600rpm for 25\u2009min using 4\u2009mL of Lymphocyte Separation Medium (Fischer). For IEL preparations, faecal material was flushed from lumen of small intestine with ice-cold PBS using a gavage needle. Fatty tissues, vasculature and Peyer\u2019s patches were removed, followed by longitudinal opening and 60\u2009min agitation in RPMI-1640 10% Newborn calf serum (NCS) (Life technologies) with 5\u2009mM EDTA (Life technologies) at 37\u2009\u00b0C. Cells were subsequently passed through an autoclaved column containing 0.7\u2009g of nylon wool , equilibrated with RPMI-1640 with 38\u2009mM HEPES (Life technologies). IELs were enriched on a discontinuous Percoll gradient (40%/80% isotonic Percoll), before use and stained as described below.Fluorochrome-conjugated antibodies (eBioscience or BD) were; CD3\u03b5 (145-2C11), CD4 (RM4-5), CD8\u03b1 (53\u20136.7), CD25 (PC61), CD44 (IM7), TCR\u03b4 (GL3), TCR\u03b2 (H57\u2013597), CD8\u03b2 (H35\u201317.2), CD5 (53\u20137.3), CD69 (H1.2F3), V\u03b12 (B20.1), CD117 (2B8), B220 (RA3\u20136B2). For surface staining, cells were Fc-blocked and stained with antibodies in FACS buffer for 30\u2009mins. Cells were then washed twice with FACS buffer using a centrifugation speed of 300\u2009g for 5\u2009mins. After staining, cells were resuspended in FACS buffer containing 0.5\u2009\u03bcg/ml DAPI (Life technologies) for dead cell exclusion. For intracellular staining, cells were first stained for extracellular markers and subsequently fixed and permeabilized using the eBioscience intracellular flow kit according to the manufacturer\u2019s protocol. The corresponding isotype control was used as a negative control. Samples were acquired using an LSR-II and analysed using Flow Jo v10.Total RNA was extracted using the RNeasy Mini kit (QIAGEN) according to the manufacturer\u2019s instructions. Concentration and purity was determined using the NanoDrop ND-1000 spectrophotometer (Thermo Scientific). Total RNA was reverse-transcribed into cDNA using the High-Capacity RNA-to-cDNA\u2122 Kit (life technologies). Quantitative real-time PCR was performed using the power SYBR green PCR Master mix (life technologies) on the 7500 real-time PCR system (life technologies). Primers were designed manually and recognised intron/exon boundaries;aFwd: 5\u2032-GGCTCTACCATCAGGCATCGC-3\u2032;pT\u03b1aRev: 5\u2032-GGTGGTTTGCCTGGTCCTCG-3\u2032;pT\u03b1bFwd: 5\u2032-GCTCTACCATCAGGGGGAATCTTC-3\u2032;pT\u03b1bRev: 5\u2032-CGGGGGGACACAGCGG-3\u2032.pT\u03b1All samples were run in triplicate and expression was normalised against the housekeeping gene GAPDH. Analysis of PCR results was performed on 7500 real-time software v2.0.6 (life technologies).Cells were individually FAC-sorted into 96 well-plates containing 10\u2009\u03bcl of reverse transcription mix , 30U MMLV-RT reverse\u00a0transcriptase (life Technologies), 1% BSA and 0.5\u2009\u03bcM of the following primers;pT\u03b1geneF: 5\u2032-TAGGACATGGCTGCTGCTGC-3\u2032;pT\u03b1geneR: 5\u2032-TCCCACCCACAGAATTTGGAC-3\u2032.2 (NEB) and 0.25\u2009mM of the pT\u03b1geneF and pT\u03b1geneR primers. The second round PCR was performed using 2.0\u2009\u03bcl of the PCR product generated by the first-round PCR and consisted of a 3\u2009min denaturation step followed by 25 cycles of amplification in a PCR mix containing 6.5U of Taq polymerase, 1.6\u2009mM MgCl2, 0.25\u2009mM dNTPs, and 0.25\u2009mM of the following primers;Plates were incubated for 2hr at 37\u2009\u00b0C in a thermocycler (Biorad) and the reaction was stopped by 10\u2009min incubation at 70\u2009\u00b0C. Two rounds of PCR were then carried out on the cDNA. The first round of PCR consisted of denaturation at 94\u2009\u00b0C, followed by 34 cycles of amplification in a PCR mix containing 7.5U of Taq polymerase (NEB), 0.25\u2009mM dNTPs (NEB), 1.6\u2009mM MgClaF: 5\u2032-GCTCTACCATCAGGCATCGC-3\u2032;pT\u03b1aR: 5\u2032-CTCCAGCTGTCAGGGGAATC-3\u2032;pT\u03b1bF: 5\u2032-GCTCTACCATCAGGGGAATCTTC-3\u2032;pT\u03b1bR: 5\u2032-GCAGGTACTGTGGCTGAGC-3\u2032.pT\u03b1PCR products were run on a ReadyAgarose 96 Plus gel (Biorad) and PCR products were sequenced for validation.Data are presented as mean\u2009\u00b1\u2009s.d. A student\u2019s t-test was used to assess statistical significance between groups. A difference was considered significant if p\u2009<\u20090.05.Supplementary Figures"} +{"text": "It has been highlighted that the original article , publishChanges:A labeling error in Table\u00a0[Without DPN n\u2009=\u2009607 (100%)] was replaced by (Yes n\u2009=\u2009396).[With DPN n\u2009=\u2009396 (100%)] was replaced by (No n\u2009=\u2009607).[Age group (year): (mean\u2009\u00b1\u2009SD)] was replaced by [Age (year)], and the mean age was removed from the table.2): (mean\u2009\u00b1\u2009SD)] was replaced by [Body mass index (kg/m2)] and the mean BMI was removed.[Body mass index (BMI) (kg/m[Duration of diabetes (year): (mean\u2009\u00b1\u2009SD)] was replaced by [Duration of diabetes (year)] and the mean duration was removed.SBP mmHg (mean\u2009\u00b1\u2009SD) DBP mmHg (mean\u2009\u00b1\u2009SD) was removed.All these were removed TC (mean\u2009\u00b1\u2009SD), LDL (mean\u2009\u00b1\u2009SD), HDL (mean\u2009\u00b1\u2009SD).[HbA1C (%): (mean\u2009\u00b1\u2009SD)] was replaced by [HbA1c (%)], and the mean HbA1c was removed from the table."} +{"text": "Their crystal structures exhibits alternating inorganic and organic stacking sheets or layers in (II), with Cl 6H22N4)[SnCl6]Cl2\u00b72H2O, (I), and 1,4-bis\u00ad(2-ammonio\u00adeth\u00adyl)piperazin-1,4-diium hexa\u00adchlorido\u00adstannate (IV) dichloride dihydrate, (C8H24N4)[SnCl6]Cl2\u00b72H2O, (II), have been synthesized from the same starting materials. In each case both the cations and anions are located about inversion centers. Their crystal structures exhibits alternating inorganic and organic stacking sheets in (I) and layers in (II), with Cl\u2212 ions and water mol\u00adecules occupying the space in between. The cohesion of the three-dimensional frameworks are governed by N\u2014H\u22efCl, N\u2014H\u22efO, C\u2014H\u22efCl and O\u2014H\u22efCl hydrogen bonds. Hirshfeld surface analysis of both crystal structures indicates that the H\u22efCl/Cl\u22efH contacts exert an important influence on the stabilization of the packing.Two new organic\u2013inorganic hybrid compounds, tri\u00adethyl\u00adene\u00adtetra\u00adammonium hexa\u00adchlorido\u00adstannate (IV) dichloride dihydrate, (C Piperazine derivatives are relatively more volatile than the corresponding linear polyethyl\u00adene amines and other branched or cyclic TETA, with close boiling points, such as tris-(2-amino\u00adeth\u00adyl)amine), 1,4-bis\u00ad(2-amino\u00adeth\u00adyl)piperazine, (Bis AEP), and 2 salt. Under very mild reaction conditions, we believe that (Bis AEP) is present as an impurity in commercial TETA based on the fact that rearrangement reactions of aliphatic chelating polyamines require high pressure and temperature 4+ cation, one half of an inorganic [SnCl6]2- dianion, one Cl\u2212 ion and one mol\u00adecule of water dianion [SnCl6]2\u2212, lying on a centre of inversion, exhibits a nearly perfect octa\u00adhedral coordination sphere with Sn\u2014Cl bond lengths ranging from 2.4114\u2005(6) to 2.4469\u2005(6)\u2005\u00c5 and Cl\u2014Sn\u2014Cl bond angles between 88.94\u2005(2) and 91.06\u2005(2)\u00b0.The asymmetric unit of (I)er Fig.\u00a01. The 2\u2212dianion lying on a centre of inversion to 2.4331\u2005(6)\u2005\u00c5 and Cl\u2014Sn\u2014Cl bond angles ranging from 88.55\u2005(2) to 91.45\u2005(2)\u00b0 for the cis angles [180\u00b0 for trans angles]. The organic part is totally protonated and the piperazinium portion adopts a chair conformation, with both ammonio\u00adethyl groups being in equatorial positions.The asymmetric unit of compound (II)on Fig.\u00a02. The [Bi4+ and inorganic [SnCl6]2\u2212 sheets extending along the a-axis direction. The organic cations in adjacent chains are oriented in opposite directions, forming anti\u00adparallel sheets. The isolated chloride ions Cl\u2212 and the water mol\u00adecules are located in the otherwise empty space between the sheets ts Fig.\u00a03.W and C\u2014H\u22efCl hydrogen-bonding inter\u00adactions (Table\u00a013+ group as well as the NH2+ group of [TETA]4+ act as hydrogen-bond donors. The D\u22efA distances for the NH3+ group range from 2.980\u2005(4) to 3.255\u2005(3)\u2005\u00c5, while D\u22efA distances of 3.026\u2005(2) to 3.452\u2005(2)\u2005\u00c5 are found for the NH2+ group. The water mol\u00adecules play an important role in stabilizing the crystal packing of (I)\u2212 anions and [SnCl6]2\u2212 dianions via O1W\u2014H1W\u22efCl4 and O1W\u2013-H2W\u22efCl2 hydrogen bonds with a H\u22efCl distances of 2.60\u2005(5) and 2.82\u2005(5)\u2005\u00c5, respectively. Additionally, by playing the role of acceptors, the water mol\u00adecules link the inorganic moieties with the organic cations through N1+\u2014H1B\u22efO1W and N1+\u2014H1C\u22efO1W charge-assisted hydrogen bonds with H\u22efO distances of 2.09 and 2.25\u2005\u00c5, respectively.The crystal packing of (I)s Table\u00a01. The NH34+ cations, are joined to their adjacent water mol\u00adecules through strong OW\u2014H\u22efCl hydrogen bonds, leading to a hydrogen-bonding pattern with a +\u2014H1B\u22efO1W and C6\u2014H5B\u22efCl4 hydrogen bonds, promote the formation of sheets of cations aligned parallel to the to 3.431\u2005(2)\u2005\u00c5 and the C\u22efCl distances of 3.715\u2005(3)\u2005\u00c5.In (II)e Table\u00a02. These sCrystalExplorer software In compounds (I)et al., 2016i.e. tri\u00adethyl\u00adene\u00adtetra\u00adminium bis\u00ad(sulfate) monohydrate, (C6H22N4)SO4\u00b7H2O (III), and bis\u00ad(2-ammonio\u00adeth\u00adyl)piperazin-1,4-ium tetra\u00adperchlorate tetra\u00adhydrate, (C8H24N4)4ClO4\u00b74H2O (IV), have been reported and tetra\u00adethyl\u00adene\u00adtetra\u00adamine in an HCl-acidified medium with a stoichiometric ratio of 1:1 was refluxed for one\u2005h at 343\u2005K for (I)All chemicals were used without further purificationUiso(H) = 1.2Ueq. H atoms of the water mol\u00adecule were located in a difference-Fourier map and refined with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018001044/tx2003sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 1817660, 1817659CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The planar deca\u00adchloro\u00adcyclo\u00adpenta\u00adsilane rings in the title compounds are coordinated by two chloride ions to generate inverse-sandwich complexes. n-butyl\u00adammonium) dichloride deca\u00adchloro\u00adcyclo\u00adpenta\u00adsilane di\u00adchloro\u00admethane disolvate, 2C16H36N+\u00b72Cl\u2212\u00b7Si5Cl10\u00b72CH2Cl2, (I), and bis\u00ad(tetra\u00adethyl\u00adammonium) dichloride deca\u00adchloro\u00adcyclo\u00adpenta\u00adsilane di\u00adchloro\u00admethane disolvate, 2C8H20N+\u00b72Cl\u2212\u00b7Si5Cl10\u00b72CH2Cl2, (II), both of which crystallize with discrete cations, anions, and solvent mol\u00adecules. In (I), the complete deca\u00adchloro\u00adcyclo\u00adpenta\u00adsilane ring is generated by a crystallographic twofold rotation axis. In (II), one cation is located on a general position and the other two are disordered about centres of inversion. These are the first structures featuring the structural motif of a five-membered cyclo\u00adpenta\u00adsilane ring coordinated from both sides by a chloride ion. The extended structures of (I) and (II) feature numerous C\u2014H\u22efCl inter\u00adactions. In (II), the N atoms are located on centres of inversion and as a result, the ethyl\u00adene chains are disordered over equally occupied orientations.We have determined the crystal structures of two deca\u00adchloro\u00adcyclo\u00adpenta\u00adsilanes, namely bis\u00ad(tetra- The distances of the chloride ions to the Si atoms show that the chloride ions are located almost exactly above the centroid of the ring 2[Si5Cl12] (R = nBu or Et) could be harvested after storage of the reaction solution for one week at 195\u2005K in 89% and 93% yield, respectively. Both adducts are stable in the solid phase under inert conditions. However, in solution a rapid transformation of [nBu4N]2[Si5Cl12] to [nBu4N]2[Si6Cl14] and [nBu4N]2[Si7Cl16] , while [Et4N]2[Si5Cl12] is not soluble. For comparison, a 29Si CP/MAS NMR spectrum of single crystals of [nBu4N]2[Si5Cl12] was recorded .The addition of a solution of [t) Fig.\u00a04. Crystal6] Fig.\u00a05 can be ometh\u00adyl\u2014H = 0.98\u2005\u00c5 or Cmethyl\u00adene\u2014H = 0.99\u2005\u00c5 and with Uiso(H) = 1.5Ueq(Cmeth\u00adyl) or 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a04The Cl atoms of the di\u00adchloro\u00admethane solvent mol\u00adecule in (I)10.1107/S2056989017016310/hb7717sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989017016310/hb7717Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017016310/hb7717IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989017016310/hb7717sup4.pdfFig. S1. 29Si NMR spectra. DOI: 10.1107/S2056989017016310/hb7717sup5.pdfFig. S2. 29Si CP/MAS NMR spectrum. DOI: 1585194, 1585193CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N,N\u2032,N\u2032\u2032-donating imine ligand led to a mononuclear compound, the N,N\u2032,O-donating imine ligand produced a binuclear metal complex.It is shown that tridentate imine ligands can control the nuclearity of copper(II) complexes based on the donor atoms present in the ligand. While the N,N\u2032,N\u2032\u2032-donating imine ligand led to a mononuclear compound, namely di\u00adchlorido\u00adcopper(II) monohydrate, [CuCl2(C10H15N3)]\u00b7H2O, 1, while the N,N\u2032,O-donating imine ligand produced a binuclear metal complex, namely \u03bc2-chlorido-di\u00adchlorido\u00ad(\u03bc2-2-{[2-(di\u00admethyl\u00adamino)\u00adethyl]imino\u00admethyl}phenolato)\u00addicopper(II) 0.11-hydrate, [Cu2(C11H15N2O)Cl3(C4H12N2)]\u00b70.11H2O, 2. The structure of 2 is a remarkable example of a binuclear copper(II) complex containing a single substituted 2-imino\u00admethyl\u00adphenolate ligand that has two copper(II) sites in square-pyramidal coordination.It is demonstrated here that tridentate imine ligands can control the nuclearity of copper(II) complexes based on the donor atoms present in the ligand. The Imines represent an inter\u00adesting class of ligands because they can be easily synthesized and fine-tuned to the desired application by introducing extra donor atoms or groups with the desired steric properties into the side chains. A limited number of binuclear copper(II) compounds containing substituted 2-imino\u00admethyl\u00adphenole ligands have been reported in the literature sites of haemocyanin and binuclear (2) copper(II) complexes with tridentate imine-containing ligands obtained by a one-pot synthetic method. The nuclearity of the complexes was shown to be directed by the different donor atoms present in the imine ligand.We describe here the crystal structures of mononuclear . The CuII cation is displaced from the least-squares plane defined by the four coordinating atoms of the square base by 0.334\u2005\u00c5. The bond lengths to these atoms are: Cu\u2014N1 = 2.060\u2005(2), Cu\u2014N2 = 1.978\u2005(2), Cu\u2014N3 = 2.058\u2005(2) and Cu\u2014Cl2 = 2.2639\u2005(8)\u2005\u00c5; the Cu\u2014Cl bond length to the apical Cl1 atom that completes the first coordination sphere is considerably longer, at 2.5013\u2005(8)\u2005\u00c5. In order to assess the coordination geometry of copper(II) more qu\u00adanti\u00adtatively, the \u03c45 index as defined by Addison et al. cations, both in a square-pyramidal coordination environment . The presence of the phenolate group in the structure of the imine ligand directs the reaction with copper(II) cations to form a binuclear coordination compound, in contrast with the mononuclear species 1 obtained when a pyridine group is present in the ligand. Atoms Cu1 and Cu2 in 2 are displaced from the least-squares plane defined by the four coordinating atoms of the square base by 0.299 and 0.170\u2005\u00c5, respectively. The distances from the central copper(II) cations to these ligating atoms are: Cu1\u2014N1 = 2.068\u2005(2), Cu1\u2014N2 = 1.959\u2005(2), Cu1\u2014O1 = 1.968\u2005(1) and Cu1\u2014Cl2 = 2.2958\u2005(5)\u2005\u00c5; Cu2\u2014N3 = 2.021\u2005(2), Cu2\u2014N4 = 2.040\u2005(2), Cu2\u2014Cl3 = 2.2501\u2005(5) and Cu2\u2014O1 = 2.004\u2005(1)\u2005\u00c5. The two Cu\u2014Cl distances to the apical Cl atoms are likewise longer, Cu1\u2014Cl1 = 2.5476\u2005(5)\u2005\u00c5 and Cu2\u2014Cl2 = 2.5938\u2005(5)\u2005\u00c5. The Cu\u22efCu distance within the binuclear complex is 3.2525\u2005(5)\u2005\u00c5. In compound 2, the \u03c45 index for Cu1 is 0.294 and for Cu2 0.260, indicating more distorted square-pyramidal coordination environments for both central copper(II) cations.The binuclear compound nt Fig.\u00a02a. The p2, a solvent-accessible void of 42\u2005\u00c53 was detected by a PLATON analysis distance of 3.393\u2005(2)\u2005\u00c5 . The partly occupied water mol\u00adecule participates in a hydrogen bond with the \u03bc2-bridging Cl2 ligand \u00adphenolate ligand were considered as analogues of 2. A total of 12 hits were found as analogues of 1, while 11 hits were found for analogues of 2, including both mono- and bis\u00ad(imino\u00admethyl)\u00adphenolate ligands. Averages of selected bond lengths and the statistical analysis module in Mercury (Version 3.9) compounds 1 is associated with the nonhydrated analogue , Cu\u2014N2 = 2.104\u2005(2) and Cu\u2014N3 = 2.236\u2005(2)\u2005\u00c5, and almost identical Cu\u2014Cl1 = 2.2573\u2005(5) and Cu\u2014Cl2 = 2.22561\u2005(6)\u2005\u00c5 distances. The CuII cation is displaced from the mean plane defined by the four coordinating atoms of the square base by 0.622\u2005\u00c5. While for 1 \u03c45 = 0.0593, for the structure of TAWMEK \u03c45 = 0.302. The differences in the coordination environment of copper(II) probably arise as a consequence of the presence of the hydrogen-bonded network established between the chloride ligands and the water mol\u00adecules in the crystal structure of 1. The coordination spheres around the CuII cations in 1 and TAWMEK are compared in Fig.\u00a04The closest relation to 2, the search returned only two examples of binuclear CuII complexes containing a single \u03bc2-(mono\u00adimino\u00admethyl)\u00adphenolate ligand [VAMJIE complex with a single \u03bc2-(mono\u00adimino\u00admethyl)\u00adphenolate ligand that has two CuII co\u00adordination sites in square-pyramidal environments. The co\u00adordination spheres around the two CuII cations in 2 and UFATEB are compared in Fig.\u00a04Regarding the binuclear compound N,N-di\u00admethyl\u00adethylene\u00addi\u00adamine, pyridine-2-carbox\u00adaldehyde and salicyl\u00adaldehyde were purchased from Sigma\u2013Aldrich and used without further purification.Copper(II) chloride dihydrate was purchased from Vetec (Brazil). 1, C10H15Cl2CuN3\u00b7H2O, was obtained as follows. In a 10\u2005ml beaker, N,N-di\u00admethyl\u00adethylene\u00addi\u00adamine was combined with pyridine-2-carbox\u00adaldehyde in methanol (200\u2005\u00b5l). The reaction was carried out at room temperature for 24\u2005h. Afterwards, solid CuCl2\u00b72H2O was added to the reaction mixture. A polycrystalline green compound was obtained, filtered off and washed with small amounts of cold methanol. Elemental analysis was performed on a Perkin\u2013Elmer CHNS-O 2400. Analysis, calculated for C10H15Cl2CuN3\u00b7H2O: C 36.4, H 5.2, N 12.7%; found: C 36.8, H 4.8, N 13.0%. The supernatant was transferred to an amber flask and green crystals suitable for single-crystal X-ray diffraction were obtained by slow evaporation.Compound 2 (C15H27Cl3Cu2N4O\u00b70.11H2O) was obtained following the same synthetic procedure as used for 1, but replacing pyridine-2-carbox\u00adaldehyde by salicyl\u00adaldehyde (11\u2005\u00b5l). Green needle-like crystals of 2 were obtained by slow evaporation of the supernatant. Since only a few crystals were obtained, no further analytical data were acquired.Compound 2) or 0.95\u2005\u00c5 (CH), with Uiso(H) = 1.2Ueq(C), and C\u2014H = 0.98\u2005\u00c5 (CH3) and Uiso(H) = 1.5Ueq(C). For structure 1, the H atoms of the water mol\u00adecule were refined with an O\u2014H distance restraint of 0.82\u2005(1)\u2005\u00c5 and a H\u22efH separation of 1.29\u2005(2)\u2005\u00c5, and with Uiso(H) = 1.5Ueq(O). For structure 2, the H atoms of the amine functionality (H3A and H3B) were refined freely. The occupancy of the partly occupied water solvent mol\u00adecule was refined to a value of 0.11\u2005(1); for this mol\u00adecule, H atoms were not located and they were not considered in the final model.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989017013652/wm5417sup1.cifCrystal structure: contains datablock(s) global, 2, 1. DOI: 10.1107/S2056989017013652/wm54171sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989017013652/wm54172sup3.hklStructure factors: contains datablock(s) 2. DOI: 1576100, 1576099CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-tetra\u00adzol-1-olate) dianion.In energetic materials, the crystal density is an important parameter that affects the performance of the material. When making ionic energetic materials, the choice of counter-ion can have detrimental or beneficial effects on the packing, and therefore the density, of the resulting energetic crystal. Presented herein are a series of five ionic energetic crystals, all containing the 5,5\u2032-bis\u00ad(1 H-tetra\u00adzol-1-olate), with the following cations: hydrazinium (1) (2N2H5+\u00b7C6N12O42\u2212), hydroxyl\u00adammonium (2) 2NH4O+\u00b7C6N12O42\u2212 , di\u00admethyl\u00adammonium (3) (2C2H8N+\u00b7C6N12O42\u2212), 5-amino-1H-tetra\u00adzol-4-ium (4) (2CH4N5+\u00b7C6N12O42\u2212\u00b74H2O), and amino\u00adguanidinium (5) (2CH7N4+\u00b7C6N12O42\u2212). Both the supra\u00admolecular inter\u00adactions and the sterics of the cation play a role in the density of the resulting crystals, which range from 1.544 to 1.873 Mg\u2005m\u22121. In 5, the tetra\u00adzolate ring is disordered over two positions [occupancy ratio 0.907\u2005(5):0.093\u2005(5)] due to a 180\u00b0 rotation in the terminal tetra\u00adzole rings.In energetic materials, the crystal density is an important parameter that affects the performance of the material. When making ionic energetic materials, the choice of counter-ion can have detrimental or beneficial effects on the packing, and therefore the density, of the resulting energetic crystal. Presented herein are a series of five ionic energetic crystals, all containing the dianion 5,5\u2032-bis\u00ad(1 As a result of the variety of cation structures and inter\u00admolecular inter\u00adactions, each exhibits subtly different crystal packing, which affects the resulting density. The mol\u00adecule of inter\u00adest, however, only exhibits minor changes in bond distances depending on the cation.One of the critical parameters directly related to the performance of an energetic material, specifically its detonation velocity, is the density of the material (Ma H-tetra\u00adzol-1-olate), is comprised of four penta\u00adnuclear rings, with two 1,2,4-oxa\u00addiazole rings linked together through the 5-position carbon atom, and the tetra\u00adzol-1-olate rings linked at the 5-position carbon atom to each 1,2,4-oxa\u00addiazole ring at the 3-position carbon.The primary mol\u00adecule, 5,5\u2032-bis\u00ad%] in which one tetra\u00adzolate is flipped by 180\u00b0. The N4\u2014C5\u2014C6\u2014N10 torsion angles for 1 [174.25\u2005(13)\u00b0], 4 , and 5 show only slight deflections from coplanar, while in 2 [168.63\u2005(15)\u00b0], the deflection is more pronounced. In structure 3, the N4\u2014C5\u2014C6\u2014N10 dihedral angle is 2.38\u2005(19)\u00b0, showing only a slight deviation from coplanarity, despite the proximity of the two electronegative oxygen atoms.In each structure, the oxa\u00addiazole oxygen atoms are on opposite sides. For 4 Fig.\u00a04, with thet al., 1987et al., 1987In all five structures, the tetra\u00adzolate C\u2014N and N\u2014N bond distances suggest a delocalized aromatic system rather than discrete single and double bonds \u2005\u00c5 matches the distance of 1.45\u2005\u00c5 seen in hydrazinium chloride \u2005\u00c5 matches the distance of 1.41\u2005\u00c5 seen for hydroxyl\u00adammonium perchlorate and 1.4780\u2005(17)\u2005\u00c5 are consistent, albeit on the low side, with those reported for di\u00adalkyl\u00adammonium ions, on average 1.494\u2005(16)\u2005\u00c5 ]: C\u2014Namino, 1.320\u2005(2) and 1.314\u2005(2)\u2005\u00c5 (1.308\u2005\u00c5); C\u2014Nring, 1.334\u2005(2) to 1.338\u2005(2)\u2005\u00c5 (1.334 to 1.342\u2005\u00c5); C\u2014N(H)\u2014N=N, 1.357\u2005(2) to 1.366\u2005(2)\u2005\u00c5 (1.363 to 1.366\u2005\u00c5); N(H)\u2014N=N\u2014N(H), 1.272\u2005(2) and 1.269\u2005(2)\u2005\u00c5 ]: C\u2014NH2, 1.309\u2005(3) and 1.320\u2005(3)\u2005\u00c5 (1.312 and 1.320\u2005\u00c5); C\u2014N(H)(NH2), 1.337\u2005(3)\u2005\u00c5 (1.328\u2005\u00c5); and N(H)\u2014NH2, 1.420\u2005(3)\u2005\u00c5 bis\u00ad(1H-tetra\u00adzol-1-olate) (dianion) with the N2\u2014N3 bond of one dianion over the tetra\u00adzolate ring of the dianion in the neighboring column . Hydrazinium ions occupy the gaps between neighboring coplanar dianions along the b-axis, above the plane of the mol\u00adecules. One hydrazinium forms a hydrogen-bonded network linking the neighboring intra\u00adsheet dianions through the tetra\u00adzolate oxygen, tetra\u00adzolate N4 atom, and the NH3 portion of hydrazinium. Additionally, hydrogen bonds form between the NH2 portion of hydrazin\u00adium, the tetra\u00adzolate oxygen atom, and the tetra\u00adzolate N3 atom of neighboring dianions. An additional hydrogen bond connects the NH3 of one hydrazinium with the NH2 portion of the symmetry-related hydrazinium ion and 4.01\u2005(2)\u2005\u00c5. The tetra\u00adzolate oxygen atom forms an anion\u2013\u03c0 inter\u00adaction with the oxa\u00addiazole ring of a neighboring dianion, with an O1-to-centroidC6/O7/N8/C9/N10 close contact of 2.98\u2005(2)\u2005\u00c5 at an O1\u2013centroidC6/O7/N8/C9/N10\u2013O7 angle of 92.3\u2005(2)\u00b0 . Additionally, the opposing columns are staggered with respect to one another. The hydroxyl\u00adammonium cations occupy the space formed where three dianion columns meet, above the dianion planes. The arrangement of the dianions in the peaks and troughs of the packing is dictated by the hydrogen bonds between the hydroxyl\u00adammonium hydroxyl group and the tetra\u00adzolate oxygen atom, and those between the hydroxyl\u00adammonium NH3 group and O1, N2, and N4 of three symmetry-related dianions \u2005\u00c5, centroidN1\u2013N4/C5\u2013centroidC6/O7/N8/C9/N10\u2013O7 angle, 80\u2005(2)\u00b0] inter\u00adaction between the tetra\u00adzolate and oxa\u00addiazole rings. Additionally, the tetra\u00adzolate oxygen atom does not participate in an anion\u2013\u03c0 inter\u00adaction with the oxa\u00addiazole ring due to the stronger \u03c0\u2013\u03c0 inter\u00adaction. The oxa\u00addiazole rings of neighboring dianions are far apart, at a centroidC6/O7/N8/C9/N10\u2013centroidC6/O7/N8/C9/N10 distance of 4.26\u2005(2)\u2005\u00c5 and a centroidC6/O7/N8/C9/N10\u2013centroidC6/O7/N8/C9/N10\u2013N10 angle of 50\u2005(2)\u00b0, suggesting minimal \u03c0\u2013\u03c0 inter\u00adaction.Structure ns Fig.\u00a07a. Addit3, space group Pa), with the dianion stacked in a staggered arrangement, with the tetra\u00adzolate ring of one dianion over the central oxa\u00addiazole\u2013oxa\u00addiazole C\u2014C bond of the dianions above and below. The oxa\u00addiazole ring resides over the tetra\u00adzolate\u2013oxa\u00addiazole C\u2014C bond in the dianions above and below. The void space between the dianion columns is occupied by di\u00admethyl\u00adammonium ions, located within the plane of the mol\u00adecules in an up\u2013down arrangement. Two di\u00admethyl\u00adammonium ions are positioned between the sheets, forming hydrogen bonds between the NH2 group and the tetra\u00adzolate oxygen atoms of dianions in neighboring sheets and 3.99\u2005(2)\u2005\u00c5 (the latter distance to the inversion-related oxa\u00addiazole of the same dianion).Structure rn Fig.\u00a08a, with 4, space group P21/c, packs in the sheet-like pattern consisting of extended sheets containing the dianion, cations, and incorporated water . The 5-amino-1H-tetra\u00adzol-4-ium cations and water mol\u00adecules surround each dianion, isolating the dianion from other dianions within the sheets. Between the sheets, the dianion only inter\u00adacts with another dianion via one terminal tetra\u00adzolate ring, with the oxygen atom of the tetra\u00adzolate over the C\u2014C bond between the tetra\u00adzolate and oxa\u00addiazole rings. Within each sheet, there is extensive hydrogen bonding between the dianions, 5-amino-1H-tetra\u00adzol-4-ium, and incorporated water mol\u00adecules, isolating the dianions from one another in the sheet plane \u2005\u00c5 [N\u2013-centroidN1\u2013N4/C5\u2013centroidN1\u2013N4/C5 angle 62.0\u2005(2)\u00b0]. The C11-oxa\u00addiazole engages in a \u03c0\u2013\u03c0 inter\u00adaction with its symmetry equivalent as well, at a centroidC11/N12/O13/C14/N15\u2013centroidC11/N12/O13/C14/N15 distance of 3.93\u2005(2)\u2005\u00c5 . A \u03c0\u2013\u03c0 inter\u00adaction is also seen between the N30-tetra\u00adzolium ring and its symmetry equivalent, at a centroidC29/N30\u2013N33\u2013centroidC29/N30\u2013N33 distance of 3.69\u2005(2)\u2005\u00c5 . Additionally, there are two anion\u2013\u03c0 inter\u00adactions, the first between O21 and the N1-tetra\u00adzolate of a neighboring dianion, and the second between O21 and the C6-oxa\u00addiazole, with an O21\u2013centroidN1\u2013N4/C5 distance of 3.33\u2005(2)\u2005\u00c5 [O21\u2013centroidN1\u2013N4/C5\u2013N2 angle 95.8\u2005(2)\u00b0] and O21\u2013centroidC6/O7/N8/C9/N10 3.02\u2005(2)\u2005\u00c5 [O21\u2013centroidC6/O7/N8/C9/N10\u2013C6 angle 76.3\u2005(2)\u00b0].Structure er Fig.\u00a09a. The 55, space group P21/n, packs in a mixing pattern, with columns containing stacked sheets consisting of the dianion coplanar with two amino\u00adguanidinium cations . Neighboring columns of sheets are rotated by 67\u00b0 with respect to one another as a result of the hydrogen bonding of the amino group of the cation with the oxygen atom of a neighboring oxa\u00addiazole ring. In fact, it is the hydrogen-bonding inter\u00adaction between the amino group of the amino\u00adguanidinium cation and the oxygen atom of the oxa\u00addiazole that directs the mixing-type packing seen in the crystal structure. The planar portion of the amino\u00adguanidinium cation inter\u00adacts via hydrogen bonds from the unsubstituted guanidinium amines to the tetra\u00adzolate oxygen atom, oxa\u00addiazole N8, and symmetry-related oxa\u00addiazole N10 atoms of one dianion, and to the tetra\u00adzolate N2 atom of a neighboring dianion \u2005\u00c5 [centroidC6/O7/N8/C9/N10\u2013centroidA\u2013N4A/C5AN1\u2013N1A angle 65.4\u2005(2)\u00b0].Structure s Fig.\u00a010a. Neigh3 < 6 < 1 < 5 < 2. Unsurprisingly, the di\u00admethyl\u00adammonium, with minimal hydrogen bonding, non-inter\u00adacting substituents, and a poor steric match for the dianion, is the least dense of the structures shown. Amino\u00adguandinium, despite significant hydrogen bonding, exhibits a lower density as well, likely due to the directionality of the hydrogen-bond donors, which directs packing of the dianions into less efficient arrangements. Hydrazinium benefits from extensive hydrogen bonding, but the orientation of the hydrazinium directs the dianions into slightly less efficient packing than the hydroxyl\u00adammonium cation, preventing the staggering of the columns that allows for improved space occupation. The 5-amino-1H-tetra\u00adzol-4-ium cation, with the second-highest density, packs very efficiently, in extended sheets with extensive hydrogen bonding, losing out to the hydroxyl\u00adammonium cation likely only due to the included water mol\u00adecules needed to fill in gaps between the dianions and cations. Hydroxyl\u00adammonium exhibits the most efficient, highest-density packing due to the directing influence and strong hydrogen-bond donating ability of the hydroxyl group, which forms a short hydrogen bond and directs the columns into a staggered arrangement, fitting the dianions slightly closer together at the point where neighboring columns meet. The range of densities, from 1.544 to 1.873\u2005g\u2005cm\u22121, shows the significant effect that matching the hydrogen-bonding abilities and sterics of the counter-ion to the primary energetic ion has on efficient packing and, by extension, the expected performance of these ionic energetics.As demonstrated above, it is the hydrogen-bonding networks that establish the crystal packing exhibited in each example, with \u03c0\u2013\u03c0 and anion\u2013\u03c0 inter\u00adactions occurring if packing allows. As shown in Table\u00a06et al., 2016H-tetra\u00adzol-1-olate). A search using 5-[3-]-1H-tetra\u00adzolate also yields no results. Searching for the ring fragments separately yielded 443 structures for 1,2,4-oxa\u00addiazole and 127 structures for tetra\u00adzol-1-olate. The closest structures to those presented herein are dimers between similar ring fragments. A search for each of the cations yields the following results: 196 structures containing hydrazinium, 99 structures containing hydroxyl\u00adammonium, 1,583 structures containing di\u00admethyl\u00adammonium, 2,230 structures containing ammonium, 17 structures containing 5-amino-1H-tetra\u00adzol-4-ium, and 130 structures containing amino\u00adguanidinium.A search of the CSD -5,5\u2032-di\u00adchloroxime (6) and 5,5\u2032--5,5\u2032-di\u00adyl)bis\u00ad(1-hy\u00addroxy\u00adtetra\u00adzole) (7), have been described previously was added to a 20\u2005ml vial with water (1.5\u2005ml) and a stir bar. Hydrazine hydrate was added to the reaction mixture and heated until dissolved. Stirring was discontinued, the stir bar was removed, and the solution was allowed to cool slowly providing crystals of 1.Compound 3: In a round-bottom flask, fitted with a drying tube, was suspended chloroxime 6 in di\u00admethyl\u00adformamide (DMF) , which was then cooled in an ice\u2013water bath. Sodium azide was added in portions with stirring, and the reaction was allowed to warm to room temperature. Additional DMF (10\u2005ml) was added to the creamy mixture, and after 1.5\u2005h, the solids went into solution. At this point, complete formation of the di\u00adazidoxime was assumed, and cyclization to 1 proceeded as follows. A 1:1 mixture of diethyl ether/dioxane was added to the reaction mixture , and the solution was cooled to 273\u2005K with an ice bath. HBr or Cl2 gas was bubbled into the reaction at which time the temperature increased to 298\u2005K. Gas was added until the reaction temperature returned to approximately 278\u2005K, and the vessel was then stoppered and allowed to stir for 22\u2005h. The voluminous, white precipitate that formed (hygroscopic di\u00admethyl\u00adamonium bromide) was separated by vacuum filtration, and the filtrate was allowed to evaporate from a crystallizing dish. Upon evaporation, a white solid (3) in a yellow oil remained. The solid was separated from the oil by vacuum filtration (535\u2005mg). 3 was crystallized by heating in minimal water and slow cooling.Compound 4: Dihydrate 7 was added to a 20\u2005ml vial with water (1.5\u2005ml) and a stir bar. 5-Amino\u00adtetra\u00adzole was added to the mixture, which was then heated with stirring until dissolved. Stirring was discontinued, the stir bar was removed, and the solution was allowed to cool slowly providing crystals of 4.Compound 5: Dihydrate 7 was added to a 20\u2005ml vial with water (1.5\u2005ml) and a stir bar. Amino\u00adguanidinium H2CO3 was added to the mixture, which was then heated with stirring until dissolved. During dissolution, gas evolved, the solution became clear, followed by the formation of a tan\u2005precipitate. Heating was continued until complete dissolution, followed the removal of the stir bar, and slow cooling to provide crystals of 5.Compound 5, the tetra\u00adzolate ring is disordered over two positons (A and B) due to a 180\u00b0 rotation in the terminal tetra\u00adzole rings. The disorder has the relative ratio of 90.7\u2005(5):9.3\u2005(5). CCDC deposition numbers are as follows: 1, CCDC 1567779; 2, CCDC 1567780; 3, CCDC 1567783; 4, CCDC 1567784; 5, CCDC 1567804.Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S205698901800364X/pk2606sup1.cifCrystal structure: contains datablock(s) global, 1, 2, 3, 4, 5. DOI: 10.1107/S205698901800364X/pk26061sup9.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S205698901800364X/pk26062sup10.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S205698901800364X/pk26063sup15.hklStructure factors: contains datablock(s) 3. DOI: 10.1107/S205698901800364X/pk26064sup20.hklStructure factors: contains datablock(s) 4. DOI: 10.1107/S205698901800364X/pk26065sup8.hklStructure factors: contains datablock(s) 5. DOI: Click here for additional data file.10.1107/S205698901800364X/pk26061sup7.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901800364X/pk26062sup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901800364X/pk26063sup9.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901800364X/pk26064sup10.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901800364X/pk26065sup11.cmlSupporting information file. DOI: 1567804, 1567784, 1567783, 1567780, 1567779CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II complex with a heterocyclic pyrazole ligand, a potential candidate for metal catalysts, are reported. The CuII atom is coordinated by three O atoms and two N atoms, provided by a tridentate pyridine-2,6-di\u00adcarboxyl\u00adate, one pyrazole and one water ligand, forming a slightly distorted square-pyramidal geometry.The synthesis and crystal structure of tridentate pyridine-2,6-di\u00adcarboxyl\u00adate Cu 7H3NO4)(C3H4N2)(H2O)]\u00b72H2O, the CuII atom is coordinated by three O atoms and two N atoms, provided by a tridentate pyridine-2,6-di\u00adcarboxyl\u00adate (pdc), one pyrazole and one water ligand, forming a slightly distorted square-pyramidal geometry [range of O\u2014Cu\u2014O and O\u2014Cu\u2014N bond angles = 79.55\u2005(8)\u2013166.22\u2005(10)\u00b0]. The water mol\u00adecule is positioned at the apical position. In the crystal, the complex mol\u00adecule and the two crystallographically independent non-coordinating water mol\u00adecules are linked into a supra\u00admolecular layer structure parallel to the ab plane via O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds.In the title compound, [Cu(C H-pyrazol-1-yl)meth\u00adyl]pyridine are known to be catalysts of polyethyl\u00adene polymerization meth\u00adyl]pyridine was oxidized to pyridine-2,6-di\u00adcarboxyl\u00adate (pdc) by metal nitrate , pyrazole and water ligands. The coordination geometry around the CuII atom is a distorted square pyramid as indicated by the \u03c4 value of 0.113 and two oxygen atoms (O9 and O12 from pdc). The plane including the CuII atom is almost planar, with an r.m.s. deviation of 0.0847\u2005\u00c5 from the corresponding least-squares plane defined by the five constituent atoms. The pyrazole ring is twisted by 66.61\u2005(10)\u00b0 from the basal plane. The apical Cu1\u2014O19 bond length of 2.217\u2005(2)\u2005\u00c5 is much longer than those of the basal Cu\u2014O lengths [Cu1\u2014O9 = 2.026\u2005(2)\u2005\u00c5 and Cu1\u2014O12 = 2.058\u2005(2)\u2005\u00c5].The mol\u00adecular structure of the title compound is shown in Fig.\u00a01B\u22efO21, O20\u2014H20B\u22efO13 and O20\u2014H20A\u22efO10iii; symmetry code as in Table\u00a01A\u22efO20iv and O21\u2014H21B\u22efO20v; Table\u00a01i and O19\u2014H19A\u22efO9ii; Table\u00a01ab plane meth\u00adyl]pyridine in aceto\u00adnitrile (5\u2005ml) in a high-pressure vessel. After sealing the high-pressure vessel, the resulting solution was stirred for three days at 403\u2005K. The precipitate formed was removed by filtration, and the filtrate was washed with aceto\u00adnitrile and di\u00adchloro\u00admethane to get a dark-green powder. Single crystals of the title compound were obtained from its aqueous solution by slow evaporation of the solvent at 333\u2005K within five days.A solution of copper nitrate trihydrate in aceto\u00adnitrile (5\u2005ml) was added to a solution of 2,6-bis\u00ad[(1Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017016231/is5480sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017016231/is5480Isup2.hklStructure factors: contains datablock(s) I. DOI: 1584872CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II cation, located on a twofold rotation axis, is coordinated by two 2,4,6-tri\u00admethyl\u00adbenzoate anions, two nicotinamide ligands and a water mol\u00adecule in a distorted penta\u00adgonal\u2013bipyramidal geometry.The Cd 10H11O2)2(C6H6N2O)2(H2O)], contains one half of the complex mol\u00adecule, with the CdII cation and the coordinated water O atom residing on a twofold rotation axis. The CdII cation is coordinated in a bidentate manner to the carboxyl\u00adate O atoms of the two symmetry-related 2,4,6-tri\u00admethyl\u00adbenzoate (TMB) anions and to the water O atom at distances of 2.297\u2005(2), 2.527\u2005(2) and 2.306\u2005(3)\u2005\u00c5 to form a distorted penta\u00adgonal arrangement, while the distorted penta\u00adgonal\u2013bipyramidal coordin\u00adation sphere is completed by the two pyridine N atoms of the two symmetry-related monodentate nicotinamide (NA) ligands at distances of 2.371\u2005(3)\u2005\u00c5 in the axial positions. In the crystal, mol\u00adecules are linked via inter\u00admolecular N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds with R22(12), R33(8), R33(14), R33(16), R33(20), R33(22), R44(22), R55(16), R66(16) and R66(18) ring motifs, forming a three-dimensional architecture. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are H\u22efH (56.9%), H\u22efC/C\u22efH (21.3%) and H\u22efO/O\u22efH (19.0%) inter\u00adactions.The asymmetric unit of the title complex, [Cd(C A deficiency of this vitamin leads to loss of copper from the body, known as pellagra disease. Victims of pellagra show unusually high serum and urinary copper levels 2(C6H6N2O)2] \u00b7H2O , O1\u2014Cd1\u2014O2 [53.63\u2005(7)\u00b0], O1i\u2014Cd1\u2014O2i [53.63\u2005(7)\u00b0], O2\u2014Cd1\u2014O4 [84.47\u2005(5)\u00b0] and O2i\u2014Cd1\u2014O4 [84.47\u2005(5)\u00b0] in the basal plane around CdII cation is 363.77\u00b0 . This confirms the presence of the CdII cation with a small deviation from the basal plane. The distorted penta\u00adgonal\u2013bipyramidal coordination sphere is completed by the two pyridine N atoms (N1 and N1i) of the two symmetry-related monodentate nicotinamide (NA) ligands at distances of 2.371\u2005(3)\u2005\u00c5 in the axial positions \u00b0 in (II), 59.02\u2005(8)\u00b0 in (IV), 58.45\u2005(9)\u00b0 in (V) and 60.70\u2005(4)\u00b0 in (VI). In the TMB anion, the carboxyl\u00adate group is twisted away from the attached benzene ring, A (C2\u2013C7), ring by 60.94\u2005(18)\u00b0, while the benzene and pyridine rings [pyridine = B (N1/C11\u2013C15)], are oriented at a dihedral angle of 50.32\u2005(11)\u00b0. The four-membered ring D (Cd1/O1/O2/C1) is nearly planar with a maximum deviation of 0.0029\u2005(30)\u2005\u00c5 (for C1) from the mean plane, and it is oriented at dihedral angles of 60.98\u2005(11) and 81.91\u2005(7)\u00b0, with respect to the A and B rings.The near equalities of the C1\u2014O1 [1.249\u2005(4)\u2005\u00c5] and C1\u2014O2 [1.253\u2005(3)\u2005\u00c5] bonds in the carboxyl\u00adate groups indicate delocalized bonding arrangements, rather than localized single and double bonds. The O2\u2014C1\u2014O1 bond angle [121.7\u2005(3)\u00b0] seems to be slightly decreased than that present in a free acid [122.2\u00b0]. The O2\u2014C1\u2014O1 bond angle may be compared with the corresponding values of 123.5\u2005(2) and 120.4\u2005(2)\u00b0 in (II), 125.2\u2005(5)\u00b0 in (III), 119.2\u2005(3) and 123.8\u2005(2)\u00b0 in (IV), 123.6\u2005(3) and 119.4\u2005(3)\u00b0 in (V) and 123.86\u2005(13) and 118.49\u2005(14)\u00b0 in (VI), where the benzoate ions are coordinated to the metal atoms only monodentately in (III), and both monodentately and bidentately in (II), (IV), (V) and (VI). The Cd1 atom lies 0.0192\u2005(1)\u2005\u00c5 above of the planar (O1/O2/C1) carboxyl\u00adate group. The O1\u2014Cd1\u2014O2 angle is 53.63\u2005(7)\u00b0. The corresponding O\u2014via inter\u00admolecular N\u2014HNA\u22efONA, N\u2014HNA\u22efOC, O\u2014HW\u22efONA and C\u2014HTMB\u22efOC hydrogen bonds analysis in water (50\u2005ml) and nicotinamide in water (25\u2005ml) with sodium 2,4,6-tri\u00admethyl\u00adbenzoate in water (150\u2005ml) at room temperature. The mixture was filtered and set aside to crystallize at ambient temperature for six weeks, giving colourless single crystals . Combustion analysis: found; C, 57.07, H, 5.67, N, 7.92%. Calculated: C32H36CdN4O7 C, 57.42; H, 5.43, N, 8.34%. FT\u2013IR: 3390, 3122, 2921, 1669, 1619, 1539, 1445, 1399, 1113, 1038, 847, 731, 641\u2005cm\u22121.The title compound was prepared by the reaction of 3CdSO2 group and of the water mol\u00adecule were located in difference-Fourier maps and refined freely. The C-bound H atoms were positioned geometrically with C\u2014H = 0.93 and 0.96\u2005\u00c5 for aromatic and methyl H atoms, respectively, and constrained to ride on their parent atoms, with Uiso(H) = k \u00d7 Ueq(C), where k = 1.5 for methyl H-atoms and k = 1.2 for aromatic H-atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018001494/xu5916sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018001494/xu5916Isup2.hklStructure factors: contains datablock(s) I. DOI: 1818756CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Individual mol\u00adecules are stacked along the b-axis direction. The cohesion in the crystal structure is accomplished by C\u2014H\u22efF hydrogen bonds and additional off-set \u03c0\u2013\u03c0 inter\u00adactions , leading to the formation of a three-dimensional supra\u00admolecular network.The mol\u00adecule of the title 1,8-naphthyridine-BF The individual F\u2014B bond length is 1.364\u2005(5)\u2005\u00c5 and the Br\u2014C bond length 1.940\u2005(4)\u2005\u00c5. Compared with the mol\u00adecular structure of a related compound \u2005\u00c5, inter\u00adplanar distance = 3.6085\u2005(1)\u2005\u00c5, slippage = 0.472\u2005\u00c5; Cg2 is the centroid of the N2/C3\u2013C6/C11 ring; symmetry code: (i) \u2212x, \u2212y, 2\u00a0\u2212\u00a0z] ethanimidato](di\u00adfluoro)\u00adborate was added dropwise to an ice-cooled solution of 2,6-lutidine (1\u2005ml) and N-acetamide in anhydrous CH2Cl2 (80\u2005ml) under a nitro\u00adgen atmosphere. After the mixture had been stirred for 24\u2005h under ambient temperature, the reaction was quenched with 20\u2005ml distilled water. The aqueous layer was extracted with CH2Cl2 (3 \u00d7 50\u2005ml); the organic layer was dried with anhydrous Na2SO4 and the solvent removed under reduced pressure. The residue was purified by silica gel chromatography using CH2Cl2 as eluent to give the pure product as a bright white powder . Yellow crystals of the title compound were obtained from its CH2Cl2 solution by slow evaporation at room temperature.BFUiso(H) = 1.2Ueq(C) for aromatic and Uiso(H) = 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016016704/wm5325sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016016704/wm5325Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016016704/wm5325Isup3.cmlSupporting information file. DOI: 1510400CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II\u2013NaI metal\u2013organic framework has been synthesized and the X-ray structure has been reported.A novel three-dimensional bimetallic Cd 2(C8H4O4)2(C3H7NO)(H2O)2]n or [CdNa22(DMF)(H2O)2]n, is a new CdII\u2013NaI heterobimetallic coordination polymer. The asymmetric unit consists of one CdII atom, two NaI atoms, two 1,3-bdc ligands, two coordinated water mol\u00adecules and one coordinated DMF mol\u00adecule. The CdII atom exhibits a seven-coordinate geometry, while the NaI atoms can be considered to be penta\u00adcoordinate. The metal ions and their symmetry-related equivalents are connected via chelating\u2013bridging carboxyl\u00adate groups of the 1,3-bdc ligands to generate a three-dimensional framework. In the crystal, there are classical O\u2014H\u22efO hydrogen bonds involving the coordinated water mol\u00adecules and the 1,3-bdc carboxyl\u00adate groups and \u03c0\u2013\u03c0 stacking between the benzene rings of the 1,3-bdc ligands present within the frameworks.The title compound, [CdNa Studies incorporating alkaline metal ions into d10-MOFs with one type of bridging ligand to construct novel heterobimetallic d10-alkaline metal ion MOFs have been undertaken ] where 1,2,4-btc = benzene-1,2,4-tri\u00adcarboxyl\u00adate 3\u00b7(DMF)2\u00b7(m-H2O)2] where 1,4-bdcH2 = benzene-1,4-di\u00adcarb\u00adoxy\u00adlic acid 2]} where 1,3-bdcH2 = benzene-1,3-di\u00adcarb\u00adoxy\u00adlic acid 2]\u00b72H2O where OH-1,3-bdcH2 = 5-hy\u00addroxy\u00adbenzene-1,3-di\u00adcarb\u00adoxy\u00adlic acid 6(H2O)8] where ntcH3 = 5-nitro\u00adbenzene-1,2,3-tri\u00adcarb\u00adoxy\u00adlic acid , we explored mixed sources of ZnII/CdII\u2013NaI with this ligand. The expected products are prepared by using a direct synthetic method, mixing metal nitrate salts, 1,3-bdcH2 and NaOH (mole ratio 1:1:2) in water, methanol and DMF solvents. However, only the CdII\u2013NaI MOF product has been successfully synthesized. As part of our ongoing studies on this complex, we describe here the synthesis and crystal structure of a novel three-dimensional heterobimetallic CdII\u2013NaI MOF, [CdNa22(DMF)(H2O)2]n (I).Porous coordination polymers or metal\u2013organic frameworks (MOFs) constructed from I) crystallizes in the tetra\u00adgonal crystal system with polar P43 space group. The asymmetric unit of (I) consists of one CdII ion, two crystallographically independent Na(I) ions, two 1,3-bdc ligands, two coordinated water mol\u00adecules and one DMF mol\u00adecules, as shown in Fig.\u00a01II ion is coordinated by seven carboxyl\u00adate oxygen atoms from four different 1,3-bdc ligands with the Cd\u2014O bond distances range between 2.301\u2005(3) and 2.555\u2005(3)\u2005\u00c5 (Table\u00a01I). The 1,3-bdc mol\u00adecule is fully deprotonated and coordinated to the CdII and NaI ions in a \u03bc5-coordination mode, creating a one-dimensional heterobimetallic chain running parallel to the c axis, Fig.\u00a03a- and b-axis directions, generating a three-dimensional framework structure as shown in Fig.\u00a04The title compound (\u00c5 Table\u00a01. The Na1I), classical O\u2014H\u22efO hydrogen bonds and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions are observed and these inter\u00adactions presumably help to stabilize the frameworks. All water mol\u00adecules are shown to act as O\u2014H\u22efO hydrogen-bond donors towards the carboxyl\u00adate groups of the 1,3-bdc ligands (Table\u00a02Cg1\u22efCg2i distance of 3.588\u2005(3)\u2005\u00c5 and a dihedral angle of 3.8\u2005(4)\u00b0 .In the crystal of (s Table\u00a02. The \u03c0\u2013\u03c0I), only the three-dimensional coordination framework {[CdNa2]\u00b7[NH2(CH3)2]} has been reported 2 mol\u00adecules. In comparison, compound (I) contains coordinated H2O and DMF mol\u00adecules projecting into the framework channels. Other related three-dimensional heterobimetallic d10\u2013NaI coordination frameworks containing benzene\u00adpolycarboxyl\u00adate ligands have been published, such as [CdNa2(H2O)2]\u00b72H2O where OH-1,3-bdcH2 = 5-hy\u00addroxy-benzene-1,3-di\u00adcarb\u00adoxy\u00adlic acid 3\u00b7(DMF)2\u00b7(m-H2O)2] where 1,4-bdcH2 = benzene-1,4-di\u00adcarb\u00adoxy\u00adlic acid 6(H2O)8] where ntcH3 = 5-nitro\u00adbenzene-1,2,3-tri\u00adcarb\u00adoxy\u00adlic acid and NaOH (2.0\u2005mmol) in 10\u2005mL of distilled water was slowly dropped to a methano\u00adlic solution (10\u2005ml) of Cd(NO3)2\u00b74H2O (1.0\u2005mmol). The reaction mixture was stirred at 333\u2005K for 30\u2005min and allowed to cool to room temperature and then filtered. The filtrate was allowed to stand to slowly evaporate at ambient temperature. Colorless block-shaped crystals suitable for single crystal X-ray diffraction were obtained after three days (76% yield based on Cd).A mixture solution of 1,3-bdcHUiso(H) = 1.2Ueq(C). The coordinated DMF mol\u00adecule was found to be disordered with two sets of sites with a refined occupancy ratio of 0.382\u2005(10) and 0.618\u2005(10).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017013871/pj2046sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017013871/pj2046Isup2.hklStructure factors: contains datablock(s) I. DOI: 1576543CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Despite a growing appreciation of \u03b3\u03b4 T\u00a0cell contributions to numerous immune responses, the mechanisms that underpin their thymic development remain poorly understood. Here, using precursor/product relationships, we identify thymic stages in two distinct developmental pathways that generate \u03b3\u03b4 T\u00a0cells pre-committed to subsequent secretion of either IL-17A or IFN\u03b3. Importantly, this framework for tracking \u03b3\u03b4 T\u00a0cell development has permitted definitive assessment of TCR\u03b3\u03b4 signal strength in commitment to \u03b3\u03b4 T\u00a0cell effector fate; increased TCR\u03b3\u03b4 signal strength profoundly prohibited the development of all IL-17A-secreting \u03b3\u03b4 T\u00a0cells, regardless of V\u03b3 usage, but promoted the development of \u03b3\u03b4 progenitors along the IFN\u03b3 pathway. This clarifies the recently debated role of TCR\u03b3\u03b4 signal strength in commitment to distinct \u03b3\u03b4 T\u00a0cell effector fates and proposes an alternate methodology for the study of \u03b3\u03b4 T\u00a0cell development. \u2022CD44 and CD45RB identify two distinct thymic \u03b3\u03b4 T\u00a0cell developmental pathways\u2022Cytokine-secretion-independent identification of effector fate-committed \u03b3\u03b4 T\u00a0cells\u2022Sizable numbers of IL-17A-committed \u03b3\u03b4 T\u00a0cells express V\u03b31 and V\u03b32/3 chains\u2022Increased TCR\u03b3\u03b4 signal strength prohibits development of IL-17A-secreting \u03b3\u03b4 T\u00a0cells Sumaria et\u00a0al. identify distinct thymic pathways that generate murine \u03b3\u03b4 T\u00a0cells pre-committed to the secretion of IL-17A or IFN\u03b3. This permits assessment of TCR\u03b3\u03b4 signal strength in thymic commitment to \u03b3\u03b4 T\u00a0cell effector fate; increased TCR\u03b3\u03b4 signal strength profoundly prohibits development of all IL-17A-secreting \u03b3\u03b4 T\u00a0cells. Anti-tu\u2212CD8\u2212 double-negative (DN) 2 cells (rather than DN3 cells) (+ cells) may inherently require certain transcription factors was suggested to promote commitment to an IFN\u03b3-secreting fate , with we3 cells) . And cer Sox-13) . ClearlyThere is presently no accepted approach for stage-wise assessment of thymic \u03b3\u03b4 T\u00a0cell development. Indeed, although studies have analyzed V\u03b3 usage , acquisi\u2212 cells that expressed high CD44 but not CD45RB were committed to IL-17A secretion, but did not make IFN\u03b3, whereas cells that had upregulated CD45RB had potential to secrete IFN\u03b3 but not IL-17A that re-capitulates thymic T\u00a0cell development in\u00a0vitro and is suited to studying \u03b3\u03b4 T\u00a0cell development that occurs predominantly in the perinatal period. Indeed, E15 thymic lobes cultured for 7\u00a0days generate \u03b3\u03b4 T\u00a0cell subsets similar to those observed ex\u00a0vivo -purified the four CD24 d cells C. Sortedectively C and 2D.products C and 2D,+ and V\u03b36+ cells being linked to IL-17A production, with V\u03b31+ and V\u03b35+ cells linked to IFN\u03b3 expression in V\u03b36+ cells cells of making IL-17A express T-bet but not ROR\u03b3t. Interestingly, our CD44/CD45RB plots show overlap with CD44/Ly-6C plots suggested to identify naive-like and memory-like peripheral \u03b3\u03b4 T\u00a0cell subsets supports a model in which commitment to an IL-17A- or IFN\u03b3-secreting fate, with initial expression of corresponding \u201cmaster\u201d transcriptional regulators . These observations suggest thymic commitment of \u03b3\u03b4 progenitors to distinct effector fates is distinguishable from actual capacity to secrete cytokine.Importantly, our analyses identify two thymic pathways of\u00a0functional \u03b3\u03b4 T\u00a0cell differentiation that diverge from a common\u00a0CD24gulators , spans a+V\u03b41+ cells in the presence of Skint1 \u03b3\u03b4 T\u00a0cells in SKG mice that have severely reduced Zap-70 activity effects of increased TCR\u03b3\u03b4 signal strength mediated by anti-TCR\u03b4 antibody. Thus, activation of the ERK/MAP kinase pathway by strong TCR\u03b3\u03b4 signaling is a key limiter of progression to an IL-17A-secreting fate. As mentioned above, such strong signaling may reflect engagement of TCR ligand, as supported by complete segregation, in the prenatal thymus, of V\u03b35 pathway . Howeverignaling that depnsidered . These iAdditional details are available in C57BL/6 (B6) mice were purchased from Charles River Laboratories. All mice\u00a0were fetal (E14\u2013E17), neonatal (1\u20133\u00a0days), post-natal (4\u20138\u00a0days), or\u00a0adult\u00a0. All experiments involving animals were performed in full compliance with UK Home Office regulations and institutional guidelines.Thymic lobes from B6 mice were cultured on Nuclepore membrane filter discs (Whatman) in complete RPMI-1640 medium plus 10% fetal calf serum (FCS) for 7\u201314\u00a0days.OP9-DL1 cells were provided by J.C. Z\u00faniga-Pfl\u00fccker (University of Toronto).For detection of V\u03b35V\u03b41 and V\u03b36V\u03b41, cells were pre-stained with GL3 followed by 17D1. For i.c. cytokine staining (eBioscience), cells were stimulated with 50\u00a0ng/ml phorbol 12-myristate 13-acetate and 1\u00a0\u03bcg/ml ionomycin (Sigma) for 4\u00a0hr at 37\u00b0C. Acquisition was performed with an LSR-II or a Canto II (BD). Analysis was performed using FlowJo (Tree Star).GraphPad Prism software was used to analyze data, which are presented as mean \u00b1 SD. Two-tailed Student\u2019s unpaired t test was used when only two groups were compared, and one-way ANOVA with Tukey\u2019s test was used for multiple comparisons. Significance was determined at p \u2264 0.05.N.S. and C.L.G. performed experiments. N.S. and D.J.P. analyzed the data. B.S.-S. and D.J.P. designed the study. D.J.P. and N.S. wrote the paper."} +{"text": "III and DyIII with the ligand 2,6-di\u00adacetyl\u00adpyridine bis\u00ad(benzoyl\u00adhydrazone) have been synthesized and structurally characterized.Two isotypic complexes of Tb Ln(DAPBH2)(CH3OH)(H2O)3]Cl3\u00b72CH3OH , are isotypic. The central lanthanide ions are nine-coordinate, being ligated by three N and two O atoms from the penta\u00addentate DAPBH2 ligand, and four O atoms from the coordinated methanol mol\u00adecule and three coordinated water mol\u00adecules. The coordination geometry of the lanthanide ion is a distorted capped square anti\u00adprism. In the crystals, the various components are linked by O\u2014H\u22efCl, N\u2014H\u22efCl and O\u2014H\u22efO hydrogen bonds, forming three-dimensional supra\u00admolecular frameworks. Within the frameworks, there are C\u2014H\u22efCl and C\u2014H\u22efO hydrogen bonds and offset \u03c0\u2013\u03c0 inter\u00adactions (inter\u00adcentroid distance ca 3.81\u2005\u00c5).The title lanthanide complexes, [ The bond distances for the DyIII complex are slightly shorter than those of the TbIII complex as a result of the lanthanide contraction effect. The DAPBH2 ligand is approximately planar, and the LnIII ion lies out of the mean plane (O1/N2/N3/N4/O2) by a distance of 0.5754\u2005(3)\u2005\u00c5 for the TbIII complex and 0.5702\u2005(3)\u2005\u00c5 for the DyIII complex. The coordination of the DAPBH2 ligand to the lanthanide ion shows a bent arrangement . These coordination features are similar to those reported for the dysprosium DAPBH2 nitrate complex \u2005\u00c5, \u03b1 = 3.8\u2005(1)\u00b0, inter\u00adplanar distance = 3.483\u2005(1)\u2005\u00c5, slippages = 1.77 and 1.55\u2005\u00c5; Cg2 and Cg3 are the centroids of C2\u2013C7 and C18\u2013C23 rings, respectively, symmetry codes: (a) x, y\u00a0\u2212\u00a01, z; (b) x, y\u00a0+\u00a01, z]. The layers are linked by O\u2014H\u22efO, O\u2014H\u22efCl and N\u2014H\u22efCl hydrogen bonds, forming a three-dimensional supra\u00admolecular framework, which is reinforced by a series of C\u2014H\u22efCl and C\u2014H\u22efO hydrogen bonds , 2,6-di\u00adacetyl\u00adpyridine , and benzoyl\u00adhydrazine was refluxed for 2\u2005h. The resulting mixture was filtered. Vapour diffusion of diethyl ether into the filtrate afforded colourless plate-like crystals of the TbIII complex . The synthetic procedure for the DyIII complex is similar, starting from dysprosium chloride (yield 43%).A methanol solution (15\u2005ml) of TbClIII complex was restrained to 0.82\u2005\u00c5. Other hydrogen atoms were generated geometrically and refined with a riding model: N\u2014H = 0.88\u2005\u00c5, C\u2013H = 0.95\u20130.98\u2005\u00c5 with Uiso(H) = 1.5Ueq and 1.2 Ueq for other H atoms.Crystal data, data collection, and structure refinement details are summarized in Table\u00a0410.1107/S2056989018004103/su5429sup1.cifCrystal structure: contains datablock(s) TbDAPBH2, DyDAPBH2, global. DOI: 10.1107/S2056989018004103/su5429TbDAPBH2sup2.hklStructure factors: contains datablock(s) TbDAPBH2. DOI: 10.1107/S2056989018004103/su5429DyDAPBH2sup3.hklStructure factors: contains datablock(s) DyDAPBH2. DOI: 1828810, 1828809CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The organic mol\u00adecule is twisted with a near orthogonal relationship between the outer rings [dihedral angle = 71.79\u2005(6)\u00b0]. Supra\u00admolecular ribbons sustained by hydrogen bonding feature in the mol\u00adecular packing. 14H12BrN3O2\u00b7H2O {systematic name: N\u2032-[(1E)-1-(5-bromo-2-hy\u00addroxy\u00adphen\u00adyl)ethyl\u00adidene]pyridine-4-carbohydrazide monohydrate}, the central CN2O region of the organic mol\u00adecule is planar and the conformation about the imine-C=N bond is E. While an intra\u00admolecular hy\u00addroxy-O\u2014H\u22efN(imine) hydrogen bond is evident, the dihedral angle between the central residue and the benzene rings is 48.99\u2005(9)\u00b0. Overall, the mol\u00adecule is twisted, as seen in the dihedral angle of 71.79\u2005(6)\u00b0 between the outer rings. In the crystal, hydrogen-bonding inter\u00adactions, i.e. hydrazide-N\u2014H\u22efO(water), water-O\u2014H\u22efO(carbon\u00adyl) and water-O\u2014H\u22efN(pyrid\u00adyl), lead to supra\u00admolecular ribbons along the a-axis direction. Connections between these, leading to a three-dimensional architecture, are mediated by Br\u22efBr halogen bonding [3.5366\u2005(3)\u2005\u00c5], pyridyl-C\u2014H\u22efO(carbon\u00adyl) as well as weak \u03c0\u2013\u03c0 inter\u00adactions [inter-centroid separation between benzene rings = 3.9315\u2005(12)\u2005\u00c5]. The Hirshfeld surface analysis reveals the importance of hydrogen atoms in the supra\u00admolecular connectivity as well as the influence of the Br\u22efBr halogen bonding.In the title isonicotinohydrazide hydrate, C Recent inter\u00adest in the coordination of hydrazide Schiff base ligands arises owing to the presence of electron-donating nitro\u00adgen and oxygen atoms, allowing these to act as a multidentate ligands, and in some cases, function as supra\u00admolecular building blocks in their mol\u00adecular assemblies \u00b0], N2\u2014N1\u2014C1\u2014C10 [179.59\u2005(15)\u00b0] and C1\u2014N1\u2014N2\u2014C2 [171.14\u2005(18)\u00b0] torsion angles is consistent with an all-trans relationship in the central chain and a small twist about the N1\u2014N2 bond. The conformation about the imine-C2=N2 bond [1.292\u2005(2)\u2005\u00c5] is E. An intra\u00admolecular hy\u00addroxy-O1\u2014H\u22efN2(imine) hydrogen bond is noted, Table\u00a01The mol\u00adecular structures of the constituents of (I)a-axis direction and mediated by hydrogen-bonding inter\u00adactions. In essence, the water mol\u00adecule provides links between three organic mol\u00adecules via hydrazide-N\u2014H\u22efO(water), water-O\u2014H\u22efO(carbon\u00adyl) and water-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonds, Table\u00a014O}2 synthons as shown in Fig.\u00a02a. Lateral connections between ribbons are via halogen bonding of the type Br\u22efBr. Here, the Br\u22efBri separation is 3.5366\u2005(3)\u2005\u00c5 . The C7\u2014Br\u22efBri angle is 156.56\u2005(5)\u00b0, and, being disposed about a centre of inversion, the C7\u2014Br\u22efBri\u2014C7i torsion angle is constrained by symmetry to 180\u00b0. The geometric characteristics indicate the Br\u22efBri halogen bond is classified as a type I halogen bond within the surface near the water-O1W atoms as well as the blue regions about the water-H1W and H2W atoms, which correspond to the acceptor and donors of the hydrogen bonds, respectively. Similarly, the other donor and acceptor atoms participating in the more significant inter\u00admolecular inter\u00adactions are viewed as the blue and red regions, respectively, in Fig.\u00a03dnorm in Fig.\u00a04b also highlight the significant contribution of Br\u22efBr and C\u22efC contacts to the mol\u00adecular packing. The presence of faint-red spots near the pyridyl-N3, C13 and H13 atoms and the carbonyl-O1 atom indicate their contributions to short inter\u00adatomic C\u22efN/N\u22efC contacts and shape-index- mapped Hirshfeld surfaces highlighting the various short inter\u00adatomic contacts influential on the mol\u00adecular packing are illustrated in Fig.\u00a05a, C\u22efC and C\u22efN/N\u22efC contacts, Fig.\u00a05b, and Br\u22efBr and Br\u22efH/H\u22efBr contacts, Fig.\u00a05c, identify their roles in consolidating the packing in the crystal.The analysis of the Hirshfeld surface for (I)s Table\u00a02 and compm- Fig.\u00a05a and b x- Fig.\u00a05c mappeda, and those delineated into H\u22efH, O\u22efH/H\u22efO, C\u22efH/H\u22efC, Br\u22efH/H\u22efBr, N\u22efH/H\u22efN, C\u22efC, Br\u22efBr and C\u22efN/N\u22efC contacts hydrogen bonds are seen on the dnorm-mapped Hirshfeld surface in Fig.\u00a04b). The conformational relationship between each of the pyridyl and benzene rings to the central planar region make these residues available for forming C\u22efH/H\u22efC contacts. The significant contribution of 17.9% from C\u22efH/H\u22efC contacts results from the short inter\u00adatomic contacts listed in Table\u00a02d with the pair of peaks at de\u00a0+\u00a0di\u00a0\u223c\u00a02.8\u2005\u00c5; these short inter\u00adatomic contacts are illustrated in Fig.\u00a05a. A forceps-like fingerprint plot corresponding to Br\u22efH/H\u22efBr contacts in Fig.\u00a06e with its tips at de\u00a0+\u00a0di\u00a0\u223c\u00a03.0\u2005\u00c5 represents the influence of the halogen\u22efhydrogen inter\u00adactions in the mol\u00adecular packing. Along with Br\u22efH/H\u22efBr contacts, Table\u00a02via Br\u22efBr contacts, as evident in Fig.\u00a06h as a very thin line beginning at de\u00a0+\u00a0di\u00a0\u223c\u00a03.5\u2005\u00c5. The contributions from other inter\u00adatomic contacts involving the bromide atom have negligible effect on the crystal stability because their inter\u00adatomic distances are much greater than sum of their respective van der Waals radii. The small but notable contributions from the C\u22efC and C\u22efN/N\u22efC contacts to the Hirshfeld surface, Table\u00a02g, a spear-shaped distribution of points with the tip at de\u00a0+\u00a0di\u00a0\u223c\u00a03.2\u2005\u00c5 and an adjacent arrow-like distribution of points at de\u00a0=\u00a0di\u00a0\u223c\u00a01.9\u2005\u00c5 result, respectively, from inter\u00adatomic C\u22efC contacts and \u03c0\u2013\u03c0 stacking inter\u00adactions involving the C3\u2013C8 ring. The short inter\u00adatomic C\u22efN/N\u22efC contacts involving the pyridyl-C13 and N3 atoms, Fig.\u00a05b, are reflected in a pair of small peaks at de\u00a0+\u00a0di\u00a0\u223c\u00a03.2\u2005\u00c5 in Fig.\u00a06i. The small contributions from other inter\u00adatomic contacts listed in Table\u00a02The overall two-dimensional fingerprint plot, Fig.\u00a06et al., 2016et al., 2014cf. the near to orthogonal relationship in (I)The most closely related structure to (I)\u22121. The 1H NMR spectrum was recorded at room temperature in CDCl3 solution on a Jeol ECA 400\u2005MHz FT\u2013NMR spectrometer.All chemicals and solvents were used as purchased without purification, and all reactions were carried out under ambient conditions. The melting point was determined using an Electrothermal digital melting-point apparatus and was uncorrected. The IR spectrum for the compound was obtained on a Perkin Elmer Spectrum 400 FT Mid-IR/Far-IR spectrophotometer from 4000 to 400\u2005cmn-butyl\u00adtin dichloride in methanol (10\u2005ml). The resulting mixture was stirred and refluxed for 3\u2005h. A cloudy orange solution was obtained and the mixture was filtered. The filtrate was allowed to stand at room temperature and yellow crystals, suitable for X-ray crystallographic studies, were obtained after the slow evaporation. The yellow crystals were found to be a side-product from the reaction mixture. Yield: 0.112\u2005g, 34%; M.p. 501\u2005K. IR (cm\u22121): 3158(br), 1666(s), 1548(s), 1152 (m), 964(s) cm\u22121. 1H NMR (in CDCl3): 11.20 , 8.73-8.82, 7.92-8.20, 6.80-6.99 , 4.82 , 4.10 , 3.13 .1-(5-Bromo-2-hy\u00addroxy\u00adphen\u00adyl)ethyl\u00adidene]iso\u00adnicotino\u00adhydra\u00adzide and tri\u00adethyl\u00adamine in methanol (25\u2005ml) were added to di-Uiso(H) set to 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989017004790/hb7669sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017004790/hb7669Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017004790/hb7669Isup3.cmlSupporting information file. DOI: 1540550CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound is an epimer of natural tetra\u00adcyclic isosaxorumamide. The di\u00adhydro\u00adfuran\u00adone, oxolane, azepane and pyrrolidine rings adopt planar, twist, twist-chair and envelope forms, respectively. In the crystal, O\u2014H\u22efO hydrogen bonds connect the water and main mol\u00adecules into a tape structure, which is further expanded into a three-dimensional network by C\u2014H\u22efO inter\u00adactions. 17H23NO4\u00b7H2O, is an epimer of the natural tetra\u00adcyclic alkaloid isosaxorumamide which consists of a fused 5\u20137\u20135 tricyclic core and a di\u00adhydro\u00adfuran\u00adone substituent. The terminal di\u00adhydro\u00adfuran ring is essentially planar with a maximum deviation of 0.0273\u2005(14)\u2005\u00c5 from the mean plane and oxolane, azepane and pyrrolidine rings in the tricyclic ring system adopt twist, twist-chair and envelope forms, respectively. In the crystal, the amide and water mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds, forming a tape structure running along the b-axis direction. The tapes are further connected by C\u2014H\u22efO inter\u00adactions into a three-dimensional architecture.The title compound, C Stemona alkaloids isolated from the root of Stemona Saxorum = 0.350\u2005(3)\u2005\u00c5 and \u03c6(2) = 271.0\u2005(4)\u00b0. Atoms C8 and C9 deviate from the plane through the other three atoms by 0.309\u2005(6) and \u22120.271\u2005(6)\u2005\u00c5, respectively. The central seven-membered azepane ring (N1/C2\u2013C5/C9/C10), which is trans-fused to the oxolane ring, adopts a twist-chair form with puckering parameters of Q = 0.796\u2005(2), Q(2) = 0.472\u2005(2)\u2005\u00c5, \u03c6(2) = 195.0\u2005(3)\u00b0, Q(3) = 0.641\u2005(2)\u2005\u00c5 and \u03c6(3) = 246.7\u2005(2)\u00b0. The pyrrolidine ring (N1/C10\u2013C13) fused to the azepane ring adopts an envelope form with puckering parameters of Q(2) = 0.300\u2005(3)\u2005\u00c5 and \u03c6(2) = 251.1\u2005(5)\u00b0. The flap atom C11 deviates from the mean plane through the other four atoms by 0.473\u2005(4)\u2005\u00c5. The amide moiety (N1/C2/C10/C13/O14) is planar, and atoms N1 and O14 deviate from the mean plane through the other three atoms by 0.028\u2005(2) and \u22120.035\u2005(4)\u2005\u00c5, respectively.The asymmetric unit of the title compound is shown in Fig.\u00a02W\u2014H1WA\u22efO14i and O1W\u2014H1WB\u22efO14ii; symmetry codes as in Table\u00a01b-axis direction. A C\u2014H\u22efO inter\u00adaction graph-set motif. Furthermore, a weak C\u2014H\u22efO inter\u00adaction , i.e. the fused tricyclic core related to the title compound , with CSD refcodes VATJAC , with refcodes PROTMI is the natural alkaloid (\u2013)-stemo\u00adamide, which is a -7,13-dione derivative of (a), and XATFOP (space group P21/n) is its racemate.In the Cambridge Structural Database . HRMS (ESI) m/z calculated for C17H24NO4+ [M + H]+: 306.1705; found: 306.1703.The title compound was afforded in a synthetic study of saxorumamide and isosaxorumamide, from ethyl 4-bromo\u00adbutano\u00adate and a siloxypyrrole analogue (Yoritate Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C). Water H atoms were located in a difference-Fourier map, and then refined freely with Uiso(H) = 1.5Ueq(O), and with distance restraints of O\u2014H = 0.84\u2005(2)\u2005\u00c5 and H\u22efH = 1.33\u2005(4)\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018004425/is5494sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018004425/is5494Isup2.hklStructure factors: contains datablock(s) I. DOI: 1830268CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-(2-iodo\u00adphen\u00adyl)acetamide into twofold inter\u00adwoven sheets, and the mol\u00adecules of 2-(4-chloro\u00adphen\u00adyl)-N-(pyrazin-2-yl)acetamide are linked into complex sheets built solely from hydrogen bonds.A combination of N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds together with C\u2014Cl\u22ef\u03c0(arene) and C\u2014I\u22ef\u03c0(arene) inter\u00adactions links the mol\u00adecules of 2-(4-chloro\u00adphen\u00adyl)- N-(2-iodo\u00adphen\u00adyl)acetamide, C14H11ClINO, mol\u00adecules are linked by a combination of N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds to form a C(4)C(4)[R21(7)] chain of rings and chains of this type are linked by a combination of C\u2014Cl\u22ef\u03c0(arene) and C\u2014I\u22ef\u03c0(arene) inter\u00adactions to form deeply puckered twofold inter\u00adwoven sheets. In the crystal of 2-(4-chloro\u00adphen\u00adyl)-N-(pyrazin-2-yl)acetamide, C12H10ClN3O, mol\u00adecules are linked into complex sheets by N\u2014H\u22efN, C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds, and by C\u2014H\u22ef\u03c0(arene) inter\u00adactions.In the crystal of 2-(4-chloro\u00adphen\u00adyl)- R1CH2CONHR2, where R1 and R2 are aromatic or hetero-aromatic substituents, are of inter\u00adest as they have some resemblance to benzyl penicillins ca 0.01\u2005\u00c5, indicating that this ring is fully aromatic.In the pyrazine ring of compound (II)C(4)C(4) direction x, y, z, where Cg1 represents the centroid of the C11\u2013C16] ring, I\u22efCg 3.7977\u2005(14), C\u22efCg 5.082\u2005(3)\u2005\u00c5 and C\u2014I\u22efCg 116.34\u2005(8)\u00b0; for C24\u2014Cl24\u22efCg2ii , Cl\u22efCg 3.4557\u2005(8), C\u22efCg 4.504\u2005(3)\u2005\u00c5 and C\u2014Cl\u22efCg 116.19\u2005(11)\u00b0. The metrics of the C\u2014Cl\u22efCg inter\u00adaction are well within the normal range, as deduced using database analysis on Fig.\u00a03. There ion Fig.\u00a03, lying won Fig.\u00a03. The dimi 3.058\u2005(2)\u2005\u00c5 and C12\u2014I12\u22efO1i 170.88\u2005(8)\u00b0 which complements the C\u2014Cl\u22efCg contact. The I\u22efO distance here is significantly shorter than the sum of the van der Waals radii, 3.56\u2005\u00c5 C(5) for the group of 428 very weak reflections having Fc/Fc(max) in the range 0.000 < Fc/Fc(max) < 0.008.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016012512/su5316sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989016012512/su5316Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016012512/su5316IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989016012512/su5316Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016012512/su5316IIsup5.cmlSupporting information file. DOI: 1497360, 1497359CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N,S-chelating hydrazinecarbodi\u00adthio\u00adate ligands provide a trans-N2S2 donor set and a distorted square-planar geometry for the NiII atom. In the crystal, a three-dimensional network is sustained by C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions.Two 19H21N2S2)2], is generated by the application of a centre of inversion. The NiII atom is N,S-chelated by two hydrazinecarbodi\u00adthio\u00adate ligands, which provide a trans-N2S2 donor set that defines a distorted square-planar geometry. The conformation of the five-membered chelate ring is an envelope with the NiII atom being the flap atom. In the crystal, p-tolyl-C\u2014H\u22ef\u03c0(benzene-iPr), iPr-C\u2014H\u22ef\u03c0(p-tol\u00adyl) and \u03c0\u2013\u03c0 inter\u00adactions [between p-tolyl rings with inter-centroid distance = 3.8051\u2005(12)\u2005\u00c5] help to consolidate the three-dimensional architecture. The analysis of the Hirshfeld surface confirms the importance of H-atom contacts in establishing the packing.The complete mol\u00adecule of the title hydrazine carbodi\u00adthio\u00adate complex, [Ni(C S-R-di\u00adthio\u00adcarbazate (R = meth\u00adyl/benz\u00adyl/methyl\u00adbenz\u00adyl) and heterocyclic aldehydes or ketones have received much attention in recent years owing to their cytotoxicity S,N-chelating hydrazinecarbodi\u00adthio\u00adate anions. From symmetry, the resulting N2S2 donor set has like atoms trans, and the square-planar coordination geometry is strictly planar. Distortions from the ideal geometry are related to the deviations of angles subtended at nickel by the donor atoms, Table\u00a01i.e. \u2212165.61\u2005(17)\u00b0. Despite being involved in a formal bond to the NiII atom, the C1\u2014S1 bond length of 1.7296\u2005(19)\u2005\u00c5 is still significantly shorter than those formed by the S2 atom, i.e. C1\u2014S2 = 1.7479\u2005(18)\u2005\u00c5 and C12\u2014S2 = 1.824\u2005(2)\u2005\u00c5.The Ni2S2 donor set does not extend to the five-membered chelate ring, which has an envelope conformation with the nickel atom lying 0.465\u2005(2)\u2005\u00c5 above the least-squares plane through the remaining atoms [r.m.s. deviation = 0.0016\u2005\u00c5]. The sequence of C1=N1, N1\u2014N2 and N2=C2 bond lengths of 1.294\u2005(2), 1.408\u2005(2) and 1.300\u2005(2)\u2005\u00c5, respectively, suggests limited conjugation across this residue. Each of the benzene rings of the S- and N-bound substituents is twisted with respect to the least-squares plane through the chelate ring. Thus, a nearly orthogonal relationship exists between the chelate and p-tolyl rings, with the dihedral angle being 89.72\u2005(5)\u00b0. Less dramatic is the twist of the iPr-substituted ring with the dihedral angle being 13.83\u2005(9)\u00b0. The dihedral angle between the aromatic rings is 84.31\u2005(6)\u00b0.The planarity of the Ni.e. the S1 and N1 atoms, are involved in intra\u00admolecular inter\u00adactions, Table\u00a02i.e. p-tolyl-C\u2014H\u22ef\u03c0(iPr-benzene) and iPr-benzene-C\u2014H\u22ef\u03c0(p-tol\u00adyl) contacts, Table\u00a02p-tolyl rings self-associate via face-to-face, \u03c0\u2013\u03c0, inter\u00adactions , indicating the p-tolyl ring participates in two distinct inter\u00adactions. The result of the supra\u00admolecular association is the formation of a three-dimensional architecture, Fig.\u00a02The two sites potentially available for hydrogen bonding in (I)et al., 2017dnorm (not shown) clearly indicates the absence of conventional hydrogen bonding in the crystal. The donors and acceptors of C\u2014H\u22ef\u03c0 inter\u00adactions, involving atoms of each of the iPr-benzene and p-tolyl rings, are viewed as blue and light-red regions and correspond to the respective positive and negative potentials on the Hirshfeld surface mapped over electrostatic potential (over the range \u00b1 0.025 au), Fig.\u00a03iPr-benzene and p-tolyl rings on the Hirshfeld surface mapped over de, Fig.\u00a04i.e. \u03c0\u22efH\u2014C contacts, between iPr\u2013H11B and the p-tolyl ring are represented by red and white dotted lines, respectively in Fig.\u00a05a; the blue dotted lines in Fig.\u00a05a represent \u03c0\u2013\u03c0 stacking between p-tolyl rings at \u2212x, \u22121\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z. The other C\u2014H\u22ef\u03c0 contacts involving p-tolyl-H17 and iPr-benzene rings are illustrated in Fig.\u00a05b.The Hirshfeld surface analysis for (I)et al., 2007a\u2013f. From the qu\u00adanti\u00adtative summary of the relative contributions of the various inter\u00adatomic contacts given in Table\u00a03i.e. 95.3%. In the fingerprint plot delineated into H\u22efH contacts. Fig.\u00a06b, the points are distributed in the major part of the plot, but they do not make significant contributions to the mol\u00adecular packing as their inter\u00adatomic separations are greater than sum of their van der Waals radii, i.e. de\u00a0+\u00a0di\u00a0>\u00a02.4\u2005\u00c5. The presence of short inter\u00adatomic C\u22efH/H\u22efC contacts, see Table\u00a04i.e. 22.2%. This is consistent with the fingerprint plot, Fig.\u00a06c, where the short inter\u00adatomic C\u22efH/H\u22efC contacts appear as a pair of small peaks at de\u00a0+\u00a0di\u00a0\u223c\u00a02.8\u2005\u00c5 and also as the blue regions around the participating hydrogen atoms, namely H5 and H10B, on the Hirshfeld surface mapped over electrostatic potential, Fig.\u00a03d and e, indicate meaningful contributions to the Hirshfeld surface, Table\u00a03i.e. 2.1%, but recognizable contribution from C\u22efC contacts to the Hirshfeld surface is ascribed to \u03c0\u2013\u03c0 stacking inter\u00adactions between symmetry-related p-tolyl rings, and appear as an arrow-like distribution of points around de\u00a0=\u00a0di\u00a01.9\u2005\u00c5 in Fig.\u00a06f. The other contacts have low percentage contributions to the surface and are likely to have negligible effects on the mol\u00adecular packing, Table\u00a03The overall two-dimensional fingerprint plot and those delineated into H\u22efH, C\u22efH/H\u22efC, S\u22efH/H\u22efS and N\u22efH/H\u22efN and C\u22efC contacts , with a formal link between the two imine functionalities, the cis-N2S2 arrangement is imposed by the geometric requirements of the bis\u00ad(di\u00adthio\u00adcarbazate) di-anion di\u00adthio\u00adcarbazate complexes in the crystallographic literature (Groom S-4-methyl\u00adbenzyl\u00addithio\u00adcarbazate (S4MDTC) precursor was synthesized by following a procedure adapted from the literature : 1589, \u03bd(C=N); 997, \u03bd(N\u2014N); 823, \u03bd(C=S).The synthesized Schiff base was dissolved in hot aceto\u00adnitrile (50\u2005ml) and added to nickel(II) acetate tetra\u00adhydrate in an ethano\u00adlic solution (30\u2005ml). The mixture was heated and stirred to reduce the volume of the solution. Precipitation occurred once the mixture cooled to room temperature. The precipitate then was filtered and dried over silica gel. The complex was recrystallized from its methanol solution. Brown prismatic crystals were formed from the filtrate after being left to stand for a month. The crystals were filtered and washed with absolute ethanol at room temperature. Yield: 70%. M.p.: 479\u2013480\u2005K. Elemental composition calculated for CUiso(H) set to 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989017002419/hb7658sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017002419/hb7658Isup2.hklStructure factors: contains datablock(s) I. DOI: 1532446CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A growing body of evidence suggests that metabolic syndrome is associated with endocrine disorders including thyroid dysfunction. Thyroid dysfunction in metabolic syndrome patients may further add to cardiovascular disease risk thereby increasing mortality. This study was done to assess thyroid function in metabolic syndrome patients and evaluate its relationship with the components of metabolic syndrome.A cross sectional study was carried out among 169 metabolic syndrome patients at B P Koirala Institute of Health Sciences, Dharan, Nepal. Anthropometric measurements and blood pressure were taken. Fasting blood samples were analysed to measure glucose, triglyceride, high density lipoprotein (HDL) cholesterol and thyroid hormones .n\u2009=\u200954) metabolic syndrome patients. Subclinical hypothyroidism (26.6\u00a0%) was the major thyroid dysfunction followed by overt hypothyroidism (3.5\u00a0%) and subclinical hyperthyroidism (1.7\u00a0%). Thyroid dysfunction was much common in females than males but not statistically significant (p\u2009=\u20090.068). The relative risk of having thyroid dysfunction in females was 1.525 (CI: 0.983\u20132.368) as compared to males. Significant differences (p\u2009=\u20090.001) were observed in waist circumference between patients with and without thyroid dysfunction and HDL cholesterol which had significant negative correlation with thyroid stimulating hormone.Thyroid dysfunction was seen in 31.9\u00a0% . Metabolic syndrome constitutes a cluster of risk factors characterized by hypertension, atherogenic dyslipidemia, hyperglycemia, prothrombotic and proinflammatory conditions . This clThyroid dysfunction, prominently subclinical hypothyroidism has been observed more frequently in metabolic syndrome patients than general population . Both meThere is evidence that thyroid function may need to be assessed in patients with metabolic syndrome who are also at higher risk for CVD. Thyroid dysfunction is common in Nepal, and the prevalence of diabetes mellitus and metabolic syndrome has been rising steadily. Reports suggest that 20.7\u00a0% of the Nepalese population have metabolic syndrome based on National Cholesterol Education Program (NCEP) criteria , 9. HoweIn this study, we assessed thyroid function and examined the association between components of metabolic syndrome and thyroid function in Nepalese population with metabolic syndrome.A cross-sectional study was conducted among metabolic syndrome patients at the department of biochemistry of B P Koirala Institute of Health Sciences (BPKIHS), Dharan, Nepal from September 2013 to September 2014. One hundred sixty nine metabolic syndrome patients aged \u226520\u00a0years were selected from the hospital during the study period. The sample size was calculated by taking prevalence of thyroid dysfunction as 15\u00a0% (approximate) in this region. Metabolic syndrome was diagnosed based on modified Asian NCEP-ATP III panel criteria . The excMetabolic syndrome patients were considered to have thyroid dysfunction if patients thyroid hormones level fell outside the reference range (free T3 (4.0\u20138.3 pmol/L), free T4 (9.0\u201320.0 pmol/L) and TSH level (0.25\u20135 mIU/L)). Patients were said to be euthyroid if all thyroid hormone levels fell within reference range. Overt hypothyroidism was defined as TSH\u2009>\u20095 mIU/L and free T3\u2009<\u20094.0 pmol/L and free T4\u2009<\u20099.0 pmol/L. Subclinical hypothyroidism was considered if TSH\u2009>\u20095 mIU/L and free T3 and free T4 within reference range. Subclinical hyperthyroidism was defined as TSH\u2009<\u20090.25 mIU/L and free T3 and free T4 within reference range. The data generated from study was analyzed using SPSS version 11.0. Continuous variables were expressed as mean\u2009\u00b1\u2009SD values. Independent sample t test and one way ANOVA was applied for continuous variables and chi square test for categorical variables at 95\u00a0% confidence interval. Pearson correlation coefficients were calculated to find relationship between the components of metabolic syndrome and thyroid profile parameters at 95\u00a0% confidence interval. Relative risk with 95\u00a0% confidence interval (CI) was used to assess the risk factors for thyroid dysfunction in metabolic syndrome.n\u2009=\u200996) males and 43.2\u00a0% (n\u2009=\u200973) females, with mean age of 47\u2009\u00b1\u200912.5\u00a0years. Height, weight, body mass index (BMI), waist circumference, systolic BP and diastolic BP were 157.4\u2009\u00b1\u20098\u00a0cm, 70.7\u2009\u00b1\u20097.9 Kg, 28.6\u2009\u00b1\u20093.3 Kg/m2, 102.5\u2009\u00b1\u20096.7\u00a0cm, 129.3\u2009\u00b1\u200913.6\u00a0mmHg and 84.9\u2009\u00b1\u200911.5\u00a0mmHg respectively. Similarly, levels of biochemical parameters; fasting blood glucose, triglyceride, HDL cholesterol, free T3, free T4 and TSH were 126.2\u2009\u00b1\u200950.4\u00a0mg/dL, 198.2\u2009\u00b1\u200990.8\u00a0mg/dL, 49.9\u2009\u00b1\u200915.3\u00a0mg/dL, 5.1\u2009\u00b1\u20091.0 pmol/L, 12.0\u2009\u00b1\u20092.9 pmol/L and 4.2\u2009\u00b1\u20093.4 mIU/L respectively. Thyroid dysfunction was found in 31.9\u00a0% (n\u2009=\u200954) patients. Subclinical hypothyroidism (26.6\u00a0%) was the major thyroid dysfunction followed by overt hypothyroidism (3.5\u00a0%) and subclinical hyperthyroidism (1.7\u00a0%). Components of metabolic syndrome according to thyroid dysfunction type are shown in Table\u00a0n\u2009=\u200929) than males 26\u00a0% (n\u2009=\u200925) but not statistically significant (p\u2009=\u20090.068). Among males, 23 had subclinical hypothyroidism, 1 had overt hypothyroidism and 1 had subclinical hyperthyroidism. Similarly among females, 22 had subclinical hypothyroidism, 5 had overt hypothyroidism and 2 had subclinical hyperthyroidism. The relative risk of having thyroid dysfunction in females was 1.525 as compared to males. When metabolic syndrome parameters were compared between the patients subgroups (with TSH\u2009<\u20095 mIU/L and TSH\u2009\u2265\u20095 mIU/L), then systolic BP, diastolic BP, waist circumference, blood glucose, triglyceride and HDL cholesterol were 129.7\u2009\u00b1\u200912.6\u00a0mmHg versus 128.4\u2009\u00b1\u200916\u00a0mmHg; p\u2009=\u20090.602, 85.3\u2009\u00b1\u200911\u00a0mmHg versus 84\u2009\u00b1\u200912.8\u00a0mmHg; p\u2009=\u20090.513, 103.4\u2009\u00b1\u20096.6\u00a0cm versus 100.5\u2009\u00b1\u20096.6\u00a0cm; p\u2009=\u20090.008, 125.2\u2009\u00b1\u200953.2\u00a0mg/dL versus 128.6\u2009\u00b1\u200943.7\u00a0mg/dL; p\u2009=\u20090.692, 189.7\u2009\u00b1\u200975.5\u00a0mg/dL versus 217.9\u2009\u00b1\u2009117.6\u00a0mg/dL; p\u2009=\u20090.119 and 51\u2009\u00b1\u200916.8\u00a0mg/dL versus 47.4\u2009\u00b1\u200911\u00a0mg/dL; p\u2009=\u20090.103 respectively. Correlation between the components of metabolic syndrome and TSH and free T4 is shown in Table\u00a0p\u2009=\u20090.037) and free T4 and weak positive correlation with TSH .The study population consisted of 56.8\u00a0% between patients with and without thyroid dysfunction, and HDL cholesterol had significant negative association with TSH level, however, other components of metabolic syndrome had no significant relationships with thyroid dysfunction. Thyroid hormones affect lipid metabolism and thus the components of metabolic syndrome, and there is positive relation between TSH and LDL cholesterol, whereas negative relation between TSH and HDL cholesterol [The correlation between subclinical hypothyroidism and metabolic syndrome and its components varies in different studies and seems to be influenced by age, gender and race of study participants . In the lesterol . Our finlesterol . There alesterol . In a stlesterol . Howeverlesterol .2), had higher triglyceride levels, and an increased likelihood of having metabolic syndrome [In the present study, we observed negative association of BMI with free T3 and free T4 and weak positive correlation with TSH. In a study in Germany, euthyroid subjects with TSH in the upper normal range (2.5\u20134.5\u00a0mU/L) were more obese ."} +{"text": "Elaeodendron trichotomum (Turcz.) Lundell is reported.The crystal structure of the triterpene lactone ochraceolide A (3-oxolup-20\u2005(29)-en-30,21\u03b1-olide) isolated from 30H44O3 indeno\u00adfuran-3,11(2H)-dione], is a triterpene lactone, which was isolated from di\u00adchloro\u00admethane extract of Elaeodendron trichotomum (Turcz.) Lundell (celastraceae) stem bark. The compound has a lupane skeleton and consists of four fused six-membered rings and two five-membered rings. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO hydrogen bonds into a three-dimensional network. The configuration of ochraceolide A was proposed based on analogue compounds which belong to the lupane type.The title compound, C Kokoona ochracea (Elm.) Merril stem bark with an ED50 of 6.0\u2005\u00b5M; and hormone-dependent breast cancer with an ED50 of 9.9\u2005\u00b5M isolated from Hippocratea celastroides K. Lundell stem bark is reported and the crystal structure described.Ochraceolides A\u2013E are a group of cytotoxic lupane \u03b3-lactones isolated from the Celastraceae family. Ochraceolide A was firstly isolated from P212121 with one mol\u00adecule in the asymmetric unit and two five-membered rings (E and F). The cyclo\u00adhexane rings are trans-fused and in standard chair conformations. The cyclo\u00adpentane (C17\u2013C19/C21/C22) ring is trans-fused to the triterpene D ring and exhibits an envelope conformation [Q = 0.451\u2005(4)\u2005\u00c5 and \u03b8 = 356.7\u2005(5)\u00b0] with the puckered C17 atom having the maximum deviation of 0.285\u2005(4)\u2005\u00c5. The \u03b1-methyl\u00adene \u03b3-lactone is cis-fused at C19\u2013C21 to the cyclo\u00adpentane E ring and is essentially planar with a maximum deviation of 0.006\u2005(4)\u2005\u00c5 for atom C19. The torsion angle C20\u2014C19\u2014C21\u2014O2 is 0.8\u2005(4)\u00b0 and the weighted average absolute inter\u00adnal torsion angle for the lactone ring is 0.7\u2005(2)\u00b0The title compound has a lupane skeleton and crystallizes in the ortho\u00adrhom\u00adbic space group it Fig.\u00a01. The triA rings of adjacent mol\u00adecules inter\u00adact through two hydrogen bonds (C2\u2014H2A\u22efO2 and C24\u2014H24A\u22efO3) in a head-to-tail arrangement, forming chains along [001]. These chains are further connected through a weak hydrogen bond between the oxygen of the ketone group (O1) and a methyl\u00adene group on the C ring (C12), forming an overall three-dimensional network.In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO hydrogen bonds Table\u00a01. The lacet al., 2016H-cyclo\u00adpenta\u00ad[b]furan-2,5(3H)-dione Lundell was collected from Chunchucmil, Yucat\u00e1n, M\u00e9xico . A voucher specimen (JTun2328) was deposited at the Herbarium Alfredo Barrera Mar\u00edn, Universidad Aut\u00f3noma de Yucat\u00e1n, M\u00e9xico. Dried and milled stem bark (2100\u2005g) was exhaustively extracted by di\u00adchloro\u00admethane using a Soxhlet extraction apparatus to yield 184.2\u2005g of crude extract. A portion of the extract (100\u2005g) was chromatographed on silica gel (40\u201360\u2005\u00b5m) using a gradient elution with n-hexa\u00adne\u2013ethyl acetate (10\u2013100% ethyl acetate), to obtain 44 fractions. Single crystals suitable for X-ray structure analysis were obtained by slow evaporation of the mixture of solvents present in fractions 7\u201310 at room temperature.Uiso(H) = 1.2Ueq(C) or 1.5Ueq(methyl C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017012816/lh4023sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017012816/lh4023Isup2.hklStructure factors: contains datablock(s) I. DOI: 1573017CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-chromen-2-one), the 4-hy\u00addroxy-3-meth\u00adoxy\u00adphenyl-substituted derivative of dicoumarol, was deprotonated by the addition of tri\u00adethyl\u00adamine, yielding the respective ammonium salt which was crystallized from a methanol solution. The deprotonated dicoumarol derivative exhibits an intra\u00admolecular negative charge-assisted hydrogen bond between the deprotonated and non-deprotonated alcohol functions of the coumarol substituents.3,3\u2032-[(3-Meth\u00adoxy-4-hy\u00addroxy\u00adphen\u00adyl)methanedi\u00adyl]bis\u00ad(4-hy\u00addroxy-2 H-chromen-2-one) and tri\u00adethyl\u00adamine in methanol yielded the title compound tri\u00adethyl\u00adammonium 3-[(4-hy\u00addroxy-3-meth\u00adoxy\u00adphen\u00adyl)(4-hy\u00addroxy-2-oxo-2H-chromen-3-yl)meth\u00adyl]-2-oxo-2H-chromen-4-olate, C6H16N+\u00b7C26H17O8\u2212 or (NHEt3)+(C26H17O8)\u2212, which crystallized directly from its methano\u00adlic mother liquor. The non-deprotonated coumarol substituent shares its H atom with the deprotonated coumarolate substituent in a short negative charge-assisted hydrogen bond in which the freely refined H atom is moved from its parent O atom towards the acceptor O atom, elongating the covalent O\u2014H bond to 1.18\u2005(3)\u2005\u00c5. The respective H atom can therefore be described as being shared by two alcohol O atoms, culminating in the formation of an eight-membered ring.The reaction between 3,3\u2032-[(3-meth\u00adoxy-4-hy\u00addroxy\u00adphen\u00adyl)methanedi\u00adyl]bis\u00ad(4-hy\u00addroxy-2 The distance of the freely refined hydrogen atom to its parent atom O3 is elong\u00adated to 1.18\u2005(3)\u2005\u00c5, while the H\u22efA hydrogen-bond length to O6 is rather short at only 1.24\u2005(3)\u2005\u00c5. This inter\u00adaction is therefore the second shortest such \u2013CAHB overall and the shortest intra\u00admolecular one. In the three related deprotonated dicoumarols, the D\u22efA distances range from 2.423\u2005\u00c5 . The distances between the alcohol oxygen atoms and bound carbon atoms are 1.3005\u2005(16)\u2005\u00c5 (O3\u2014C10) and 1.2939\u2005(17)\u2005\u00c5 (O6\u2014C19); i.e. both very similar and both significantly shorter than those reported for non-deprotonated derivatives, which range from 1.331 to 1.338\u2005\u00c5 with D\u22efA = 2.7727\u2005(19)\u2005\u00c5 and D-\u2013H\u22efA = 164.5\u2005(18)\u00b0.The ammonium hydrogen atom, which was refined freely, exhibits a hydrogen bond to the carbonyl oxygen atom of the deprotonated coumarol substituent , C11\u2014C1\u2014C20 = 114.44\u2005(12), C2\u2014C1\u2014C20 = 110.79\u2005(11)\u00b0] are slightly widened compared to the ideal tetra\u00adhedral value. As this is most pronounced for the angle involving the two coumarin substit\u00aduents, it is most likely based on steric strain. The bond lengths involving the two pyran oxygen atoms are similar as observed previously, indicating conjugation between the six-membered rings in the two benzo\u00adpyran systems in a stacking fashion are observed, O\u22efO1(\u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a0D\u22efA = 2.4139\u2005(15)\u2005\u00c5] and as acceptor [O8\u22efH27B\u2014C27(\u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a0D\u22efA = 3.257\u2005(2)\u2005\u00c5] in a non-classical hydrogen bond from an amine methyl group and the non-classical donation towards O8 [C27\u2014H27B\u22efO8(\u2212x\u00a0+\u00a0y\u00a0\u2212\u00a0z\u00a0+\u00a0D\u22efA = 3.257\u2005(2) (19)\u2005\u00c5]. Supported by the hydrogen bond with the carbonyl oxygen atom O1 as acceptor [O1\u22efH8O\u2014O8(\u2212x\u00a0+\u00a0y\u00a0\u2212\u00a0z\u00a0+\u00a0D\u22efA = 2.6488\u2005(16)\u2005\u00c5], these inter\u00adactions form infinite flat chains with \u2018up and down\u2019-pointing benzo\u00adpyrane moieties protruding along b bridges the adjacent cation and anion by hydrogen bonding as a classical donor bis\u00ad(4-hy\u00addroxy-2H-chromen-2-one) was carried out by adding 1\u2005mL of tri\u00adethyl\u00adamine to its methano\u00adlic solution. The resulting transparent yellowish solution was left standing overnight to grow transparent crystals of tri\u00adethyl\u00adammonium 3-[(4-hy\u00addroxy-3-meth\u00adoxy\u00adphen\u00adyl)(4-hy\u00addroxy-2-oxo-2H-chromen-3-yl)meth\u00adyl]-2-oxo-2H-chromen-4-olate.3,3\u2032-[(3-Meth\u00adoxy-4-hy\u00addroxy\u00adphen\u00adyl)methanedi\u00adyl]bis\u00ad(4-hy\u00addroxy-2Uiso(H) = 1.5Ueq(C-methyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a02PLATON I. DOI: 10.1107/S2056989018001561/lh5866Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018001561/lh5866Isup3.cmlSupporting information file. DOI: 1818945CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Individual ions and the solvating water mol\u00adecule assemble into dimeric units located around crystallographic inversion centers 8H13N)(C17H14P)]Cl\u00b7H2O, assemble into dimeric units located around crystallographic inversion centers via N\u2014H\u22efCl and O\u2014H\u22efCl hydrogen bonds. These discrete fragments are further inter\u00adconnected into chains by C\u2014H\u22efO inter\u00adactions. The disubstituted ferrocene core in the {[1\u2032-(di\u00adphenyl\u00adphosphino)ferrocen\u00adyl]meth\u00adyl}di\u00admethyl\u00adammonium cation has an approximate synclinal eclipsed conformation and is tilted by 3.40\u2005(11)\u00b0.Individual ions and the solvating water mol\u00adecule constituting the structure of the title compound, [Fe(C The phosphine substituent at the other cyclo\u00adpenta\u00addienyl ring is oriented so that one of its pivotal P\u2014C(Ph) bonds lies nearly in the plane of the bonding five-membered ring C6\u2013C10, while the other is roughly parallel with the axis of the ferrocene unit. The angle at which the P\u2014C18 bond inter\u00adsects the C6\u2013C10 plane is 13.17\u2005(10)\u00b0, whereas the angle subtended by the P\u2014C12 bond and the Cg1\u22efCg2 line is only 8.68\u2005(5)\u00b0.The protonated amino\u00admethyl chain is directed away from the ferrocene core, with the angle between the C1\u2014N bond and the axis of the ferrocene unit, 2PfcCH2NHMe2]+ cation in the structure of the title compound is involved in an N\u2014H\u22efCl hydrogen bond to a proximal chloride anion 2Cl2(H2O)2} around the crystallographic inversion centers. These discrete units are further inter\u00adlinked into chains along the a axis via the weaker C\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions, as shown in Fig.\u00a02Each X, where X = Cl and ClO4 3SO3 was dissolved in acetic acid (10\u2005mL) and the solution was evaporated under reduced pressure. After this procedure was repeated twice using chloro\u00adform as a solvent, the residue was dissolved in a minimum amount of hot ethyl acetate. The solution was filtered and layered with hexane. Crystallization by liquid-phase diffusion over several days afforded orange crystals of the title compound. The yield was not determined.The \u2018amine\u2019 Ph25H27FeNP]Cl\u00b7H2O (481.76\u2005g\u2005mol\u22121): C 62.32, H 6.07, N 2.91%. Found: C 62.23, H 5.91, N 2.79%. ESI MS: m/z 383 ([Ph2PfcCH2]+), 428 ([Ph2PfcCH2NMe2 + H]+)Analysis calculated for [CUiso(H) set to 1.2Ueq of their bonding atom. Hydrogen atoms bonded to carbons were included in their theoretical positions and refined as riding atoms with Uiso(H) = 1.2Ueq(C).Relevant crystallographic data and structure refinement parameters are summarized in Table\u00a0210.1107/S2056989017013408/im2484sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989017013408/im2484Isup3.hklStructure factors: contains datablock(s) I. DOI: 1575391CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound provides the first crystal structure of an \u03b1-alkenyl oxa\u00adthia\u00adzolone ring. 10H7NO2S, provides the first structure of an \u03b1-alkenyl oxa\u00adthia\u00adzolone ring. The phenyl ring and the oxa\u00adthia\u00adzolone groups make dihedral angles of 0.3\u2005(3) and \u22122.8\u2005(3)\u00b0, respectively, with the plane of the central alkene group; the dihedral angle between the rings is 2.68\u2005(8)\u00b0. A careful consideration of bond lengths provides insight into the electronic structure and reactivity of the title compound. In the crystal, extended \u03c0-stacking is observed parallel to the a-axis direction, consisting of cofacial head-to-tail dimeric units [centroid\u2013centroid distance of 3.6191\u2005(11)\u2005\u00c5]. These dimeric units are separated by a slightly longer centroid\u2013centroid distance of 3.8383\u2005(12)\u2005\u00c5, generating infinite stacks of mol\u00adecules.The title compound, C In addition, the C1\u2014S1 bond [1.7379\u2005(18)\u2005\u00c5] is slightly shorter than the statistical average for mol\u00adecules of this type (1.75\u00b10.02\u2005\u00c5). Thus the pattern of bonding within the heterocycle is consistent with the Krayuskin conjugation model, leading to the hypothesis that deca\u00adrbonylation of this derivative should occur with milder conditions than observed for heterocycles substituted with saturated substituents.The title compound Fig.\u00a01 is the fThe atoms in the ring of the oxa\u00adthia\u00adzolone heterocycle form bond angles that sum to 540.0\u00b0 consistent with a planar ring . The torsion angles O1\u2014C2\u2014C3\u2014C4 [\u22122.8\u2005(3)\u00b0] and C3\u2014C4\u2014C5\u2014C6 [\u2212179.81\u2005(17)\u00b0] confirm a near planarity of the mol\u00adecule favorable for conjugation between the \u03c0-systems of the two rings and the central alkene.et al., 2002et al., 1989et al., 1999et al., 2002The planarity, bond lengths and angles in the styryl fragment are comparable with previously reported values .There are eleven crystal structures of oxa\u00adthia\u00adzolone derivatives reported in the literature \u03bbmax (log \u220a) : 228\u2005nm (4.21), 307\u2005nm (4.52).The title compound was prepared following literature methods I. DOI: 10.1107/S2056989017011264/hb7694Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017011264/hb7694Isup3.cmlSupporting information file. DOI: 1565845CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Please note that column heights and error bars in the original figures and data in the ESM tables are correct and statistical tests are valid. These corrections do not affect any results or conclusions in this article.Unfortunately, the original version of Figs.\u00a04, 5 and 6b in the article containen numbers are as follows:The correct http://media.springernature.com/full/springer-static/image/art%3A10.1186%2Fs13550-016-0218-3/MediaObjects/13550_2016_218_Fig4_HTML.gif): \u2018HCT116\u2019 (IC) n\u2009=\u20098; \u2018A549\u2019 (WWU) n\u2009=\u20097; \u2018HCT116\u2019 (WMIC) n\u2009=\u20093; \u2018H1975\u2019 (AZ) n\u2009=\u20096.Figure\u00a04 : \u2018ONU (AZ)\u2019 n\u2009=\u20096; \u2018H1975 in ONU (AZ)\u2019 n\u2009=\u20096; \u2018A431 in ONU (AZ)\u2019 n\u2009=\u20096.Figure\u00a05 : \u2018PC9\u2019 n\u2009=\u200913.Figure\u00a06b ("} +{"text": "The title compounds are differing only by the position of the chlorine atom in the benzene ring. The mol\u00adecular structures are very similar, except for the relative position of the hy\u00addroxy\u00adphenyl rings. 23H21ClN2O2, differ from each other only by the position of the Cl atom on the corresponding benzene ring: meta relative to the central sp3 C atom for (I) and para for (II). In (I), the hy\u00addroxy\u00adphenyl rings are almost parallel, the dihedral angle between the mean planes being 9.2\u2005(2)\u00b0, but in (II), the relative position of the ring is different, characterized by a dihedral angle of 48.5\u2005(1)\u00b0. Compound (I) features intra\u00admolecular O\u2014H\u22efN and inter\u00admolecular C\u2014H\u22efO hydrogen bonds, while in (II), intra\u00admolecular O\u2014H\u22efN, C\u2014H\u22efN hydrogen bonds and weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions are observed. Compound (I) was refined as an inversion twin.The title compounds, C Recent reports on a single-crystal study , C15(S). The chloro\u00adphenyl group is almost planar with atom Cl1 deviating by 0.013\u2005(1)\u2005\u00c5 from the ring in (I)The mol\u00adecular structure of the title compounds, (I)In compounds (I)C(9) chains propagating along [010]; see Fig.\u00a04C(11) chains propagating along the ab plane of the unit cell; see Fig.\u00a05In the crystal of (I)E)-({1-(3-chloro\u00adphen\u00adyl)-2-[(E)-(2-hy\u00addroxy\u00adbenzyl\u00adidene)amino]\u00adprop\u00adyl}imino)\u00admeth\u00adyl]phenol was achieved by the condensation of salicyl\u00adaldehyde (0.02\u2005mol) and 1-(3-chloro\u00adphen\u00adyl)propane-1,2-di\u00adamine (0.01\u2005mol) in ethanol . The completion of the reaction was monitored by TLC. The obtained yellow solid was purified by recrystallization from ethanol. Single crystals suitable for X-ray analysis were obtained by slow evaporation from ethanol. The above procedure was repeated with 1-(4-chloro\u00adphen\u00adyl)propane-1,2-di\u00adamine (0.01\u2005mol) instead of 1-(3-chloro\u00adphen\u00adyl)propane-1,2-di\u00adamine to synthesise 2-[(1E)-({1-(4-chloro\u00adphen\u00adyl)-2-[(E)-(2-hy\u00addroxy\u00adbenzyl\u00adidene)amino]\u00adprop\u00adyl}imino)\u00admeth\u00adyl]phenol.The synthesis of the salen ligand 2-[(1Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms. Pairs of O\u2014H bond distances were restrained to 0.82\u2005(1)\u2005\u00c5. Compound (I)Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017016292/zq2239sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989017016292/zq2239Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017016292/zq2239IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1412946, 1412945CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E conformation with respect to the C=N bond, and a dihedral angle of 14.54\u2005(11)\u00b0 between the benzene ring and the mean plane of the N\u2014N\u2014C(N)=S hydrazinecarbo\u00adthio\u00adamide unit.The mol\u00adecule of the title Schiff base, has an 8H8BrN3OS\u00b7C2H6OS, which crystallizes as a di\u00admethyl sulfoxide (DMSO) monosolvate, displays an E configuration with respect to the C=N bond, with a dihedral angle of 14.54\u2005(11)\u00b0 between the benzene ring and the mean plane of the N\u2014N\u2014C(N)=S unit. In the crystal, mol\u00adecules are linked by N\u2014H\u22efO hydrogen bonds, forming chains propagating along the b-axis direction. Within the chains there are R23(11) ring motifs, which are reinforced by C\u2014H\u22efODMSO hydrogen bonds enclosing secondary R12(6) and R23(9) loops. The chains are linked by O\u2014Hhydrox\u00adyl\u22efS hydrogen bonds, forming layers parallel to (011). Inversion-related layers are linked by short Br\u22efBr inter\u00adactions [3.5585\u2005(5)\u2005\u00c5], forming slabs parallel to (011). The inter\u00admolecular inter\u00adactions have been investigated using Hirshfeld surface studies and two-dimensional fingerprint plots. The crystal structure of the unsolvated form of the title compound has been reported previously , and its solid-state structure is compared with that of the title solvated form.The mol\u00adecule of the title Schiff base, C Schiff bases are nitro\u00adgen-containing active organic compounds that play a vital role in enzymatic reactions involving inter\u00adaction of an enzyme with a carbonyl group of a substrate \u00b0 between the benzene ring and the mean plane of the N1/N2/C8/N3/S1 unit. The C8\u2014-S1 bond distance of 1.698\u2005(2)\u2005\u00c5 is close to that expected for a C=S bond -2-thio\u00adsemicarbazone and (E)-2-[(1H-indol-3-yl)methyl\u00adene]thio\u00adsemicarbazone -1-[4-(di\u00admethyl\u00adamino)\u00adbenzyl\u00adidene]thio\u00adsemicarbazide ring motif. Comparing the two mol\u00adecules, as shown in the structural overlay of Fig.\u00a02ca. 180\u00b0 with respect to that in the unsolvated form of the mol\u00adecule. The bond lengths and bond angles of the two mol\u00adecules are similar. In the title compound, the dihedral angle between the benzene ring and the mean plane of the N\u2014N\u2014C(N)=S hydrazinecarbo\u00adthio\u00adamide unit is 14.54\u2005(11)\u00b0 compared to ca 7.05\u00b0 in the unsolvated phase. Kargar et al. ; see Fig.\u00a04x, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01) inter\u00adactions of 3.5585\u2005(5)\u2005\u00c5, forming slabs parallel to (011), as illustrated in Fig.\u00a05In the crystal, the Schiff base hydrazone is hydrogen bonded , S\u22efH/H\u22efS (18.8%), O\u22efH/H\u22efO (13.3%), Br\u22efH/H\u22efBr (11.6), C\u22efH/H\u22efC (8.8%), N\u22efH/H\u22efN (3.4%), C\u22efC (2.8%), Br\u22efN/N\u22efBr (2.0%), Br\u22efBr (1.5%), Br\u22efO/O\u22efBr (1.1%), Br\u22efC/C\u22efBr (1.1%), C\u22efN/N\u22efC (1.0%), S\u22efS (0.7%), S\u22efN/N\u22efS (0.6%) and S\u22efC/C\u22efS (0.2%), as shown in the two-dimensional fingerprint plots in Fig.\u00a07The three-dimensional et al., 2016viz. 5-bromo-2-hy\u00addroxy\u00adbenzaldehyde thio\u00adsemicarbazone -2-(2-hy\u00addroxy\u00adbenzyl\u00adidene)hydrazine\u00adcarbo\u00adthio\u00adamide hydrate] have been reported at 100\u2005K (UJIPIN) and 203\u2005K (UJIPOT and UJIPUZ) by Monfared et al. \u00b0 in the title compound.A search of the Cambridge Structural Database of thio\u00adsemicarbazide and a hot ethano\u00adlic solution of 5-bromo\u00adsalicyl\u00adaldehyde . The solution was then cooled and kept at room temperature. The precipitate that formed was filtered off and recrystallized from di\u00admethyl sulfoxide. Colourless block-like crystals, suitable for the X-ray analysis, were obtained in a few days on slow evaporation of the solvent.Uiso(H) = 1.5Ueq and 1.2Ueq for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018000233/su5414sup1.cifCrystal structure: contains datablock(s) global, I, 1. DOI: 10.1107/S2056989018000233/su5414Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018000233/su5414Isup3.cmlSupporting information file. DOI: 1587285CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports6: Article number: 37272; 10.1038/srep37272 published online: 11172016; updated: 03072017.In the methods section, under the subheading \u2018Human Fetal Brain Atlas Dataset\u201950\u201d.\u201cGenerally, this dataset includes T2 templates and tissue probability maps for ages between 23\u201337 weeks of gestation. To create the 4D probabilistic atlas, 142 T2-weighted fast-spin echo images are acquired on 3\u2009T Philips Intera system with MR sequence parameters TR\u2009=\u20091712\u2009ms, TE\u2009=\u2009160\u2009ms, flip angle 90\u00b0 and voxel sizes 0.86\u2009\u00d7\u20090.86\u2009\u00d7\u20091\u2009mm. The atlas construction is described in detail elsewhereshould read:et al.50\u201d.\u201cT2 weighted MR images from 80 fetuses with normal brain appearances were used to create the 4D atlas. The age range at the time of scan was 21.7 to 38.7 weeks gestational age (GA), with mean and standard deviation of 29.6\u2009\u00b1\u20094.6 weeks. The images were acquired on 1.5 T Philips Achieva system with the following parameters: T2 weighted single shot Fast Spin Echo (ssFSE) TR\u2009=\u200915000\u2009ms, TE\u2009=\u2009160\u2009ms, flip angle\u2009=\u200990\u00b0 and voxel size\u2009=\u20091.25\u2009\u00d7\u20091.25\u2009\u00d7\u20092.5\u2009mm. For each subject multiple stacks of images were acquired in approximately transverse, sagittal and coronal planes and the data reconstructed into a single 3D brain volume using the slice-to-volume reconstruction method. The reconstruction voxel size was 1.18\u2009\u00d7\u20091.18\u2009\u00d7\u20091.18\u2009mm. More details can be found in studies from Serag In addition, reference 50 was listed incorrectly. The correct reference appears below:et al. Construction of a consistent high-definition spatio-temporal atlas of the developing brain using adaptive kernel regression. NeuroImage59, 2255\u20132265 (2012).Serag, A."} +{"text": "This packing motif still enables significant \u03c0\u2013\u03c0 inter\u00adactions between two pyridyl groups, and may result from the close proximity of the tetra\u00adfluorido\u00adborate ions to the platinum(II) complexes, resulting in intra\u00admolecular H\u22efF distances between 2.156 and 2.573\u2005\u00c5.The crystal structure of a platinum(II) supra\u00admolecular building block, [Pt(dbbpy)(NCCH The solubility and apt geometry of the (dbbpy)platinum(II) complex make it a desirable building block for coordination-driven self-assembly of homo-metallic PtCl2 mol\u00adecule: one with Pt\u2014N distances of 2.013\u2005(2) and 2.011\u2005(2)\u2005\u00c5 and a 79.79\u2005(6)\u00b0 N\u2014Pt\u2014N angle Pt(OH2)2](OTf)2 Pt(NCCH3)(Ph)] [BAr\u20194], containing a Pt\u2014N distance of 2.000\u2005(4)\u2005\u00c5, located trans to the aceto\u00adnitrile, while the phenyl ligand causes an elongation to 2.092\u2005(4)\u2005\u00c5 for the other Pt\u2014N bond and 1.995\u2005(4)\u2005\u00c5, respectively, with a bond angle of 80.5\u2005(2)\u00b0. These are shorter than those affected by the stronger Day, 2009, and theDay, 2009. The Pt\u20142 2](OTf)2 (Ph)][BAr\u20324] phenyl; McKeown et al., 2011et al., 2003tert-butly groups for the (dbbpy)Pt(NCCH3)2 cation and its corresponding dimer are at an angle of 10.82\u00b0], both of which accommodate the bulky tert-butyl groups of the dbbpy ligands. The intra\u00admolecular Pt\u2014Pt distance is quite long at 4.5123\u2005(3)\u2005\u00c5, yet the pyridyl rings of the dbbpy are positioned for \u03c0\u2013\u03c0 inter\u00adactions with distances between 3.616\u2005(5)\u2005\u00c5 (N1\u22efN1i) and 4.032\u2005(7)\u2005\u00c5 (C4\u22efC4i) occurring between the two rings 2](OTf)2 2](ClO4)2 compounds containing the bulky dbbpy ligand pack as head-to-tail dimers, such as the aforementioned (dbbpy)PtClDay, 2009, [Pt(SO3CF3)2 4](BF4)2 Cl3)2](BF4)2[Pt(dbbpy) of Pt(dbbpy)Cl2, and 164\u2005mg (0.8425\u2005mmol) of AgBF4 was refluxed under stirring until a yellow solution formed. The solution was isolated, via cannula, from the AgCl precipitate and condensed under reduced pressure until \u223c5\u2005mL of orange solution remained. This was combined with 25\u2005ml of Et2O and the resulting precipitate was washed with 3 \u00d7 20\u2005mL Et2O to give 206.9\u2005mg (83.8% yield) of product. UV\u2013vis \u03bbmax (\u220a Lmol\u22121cm\u22121): 211 (4.6 \u00d7 104), 249 (4.2 \u00d7 104), 306 (2.0 \u00d7 104), 319 (2.4 \u00d7 104) and 346 (6.0 \u00d7 103) nm.Yellow crystals of the title compound were grown from liquid diffusion of hexa\u00adnes into a dilute acetone solution.Uiso(H) = 1.2Ueq(CH) or Uiso(H) = 1.2Ueq(CH3).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018005923/jj2198sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018005923/jj2198Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018005923/jj2198Isup3.molSupporting information file. DOI: 1837532CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A step-like conformation about the pyranyl ring is found for the mol\u00adecular structure of the title compound. The three-dimensional packing is sustained by \u03c0\u2013\u03c0, C\u2014Cl\u22ef\u03c0 and C\u2014H\u22efO inter\u00adactions. 19H13ClO3Se {systematic name: 2-[(4-chloro\u00adphen\u00adyl)selan\u00adyl]-2H,3H,4H,5H,6H-naphtho\u00adpyran-5,6-dione}, has the substituted 2-pyranyl ring in a half-chair conformation with the methyl\u00adene-C atom bound to the methine-C atom being the flap atom. The dihedral angle between the two aromatic regions of the mol\u00adecule is 9.96\u2005(9)\u00b0 and indicates a step-like conformation. An intra\u00admolecular Se\u22efO inter\u00adaction of 2.8122\u2005(13)\u2005\u00c5 is noted. In the crystal, \u03c0\u2013\u03c0 contacts between naphthyl rings [inter-centroid distance = 3.7213\u2005(12)\u2005\u00c5] and between naphthyl and chloro\u00adbenzene rings [inter-centroid distance = 3.7715\u2005(13)\u2005\u00c5], along with C\u2014Cl\u22ef\u03c0(chloro\u00adbenzene) contacts, lead to supra\u00admolecular layers parallel to the ab plane, which are connected into a three-dimensional architecture via methyl\u00adene-C\u2014H\u22efO(carbon\u00adyl) inter\u00adactions. The contributions of these and other weak contacts to the Hirshfeld surface is described.The title organoselenium compound, C It exhibits biological activities in the context of cancer can be isolated from the bark of the lapacho tree found in Central and South American countries Compounds of the bio-essential element selenium, found in amino acids such as seleno\u00adcysteine and seleno\u00admethio\u00adnine, are known to hold potential as pharmaceutical agents i.e. Cg(C2\u2013C4/C9\u2013C11)\u22efCg(C3\u2013C8)i = 3.7213\u2005(12)\u2005\u00c5 for an angle of inclination = 0.72\u2005(9)\u00b0 and symmetry operation (i) \u2212x, \u2212y, \u2212z. Two types of inter\u00adactions connect centrosymmetric aggregates into a supra\u00admolecular layer parallel to the ab plane . Thus, \u03c0\u2013\u03c0 inter\u00adactions between naphthyl and chloro\u00adbenzene rings are formed, along with C\u2014Cl\u22ef\u03c0(chloro\u00adbenzene) contacts between centrosymmetrically related rings (Table\u00a01b).In the mol\u00adecular packing of (I)ne Fig.\u00a02a. Thus,s Table\u00a01. Connects Table\u00a01 to consos Table\u00a01b.et al., 2016dnorm in Fig.\u00a03dnorm-mapped Hirshfeld surface confirms the absence of conventional hydrogen bonds in the structure except for a weak C\u2014H\u22efO inter\u00adaction as given in Table\u00a01The Hirshfeld surfaces calculated on the structure of (I)a) and those delineated into H\u22efH, O\u22efH/H\u22efO, Cl\u22efH/H\u22efCl, C\u22efC, C\u22efH/H\u22efC, C\u22efCl/Cl\u22efC and Cl\u22efO/O\u22efCl contacts de + di \u223c2.3\u2005\u00c5 in Fig.\u00a06b is the result of a short inter\u00adatomic H\u22efH contact ; the points arising from the short inter\u00adatomic O\u22efH contacts are merged in the plot.The overall two-dimensional fingerprint plot Fig.\u00a06a and tht Table\u00a02. The intot Fig.\u00a06c; the pe, characterizes the two \u03c0\u2013\u03c0 stacking inter\u00adactions, one between inversion-related naphthyl rings, and the other between the chloro\u00adbenzene and (C2\u2013C4/C9\u2013C11) rings as the two overlapping triangular regions at around de = di \u223c1.8 and 1.9\u2005\u00c5, respectively, having green points in the overlapping portion. The presence of these two \u03c0\u2013\u03c0 stacking inter\u00adactions is also seen in the flat regions around the participating rings labelled with 1, 2 and 3 in the Hirshfeld surface mapped over curvedness in Fig.\u00a07The fingerprint plot delineated into C\u22efC contacts, Fig.\u00a06i.e. 3.0%, contribution from C\u22efCl/Cl\u22efC contacts to the Hirshfeld surface is the result of its involvement in a C\u2014Cl\u22ef\u03c0 contact formed between symmetry-related chloro\u00adbenzene atoms . Its presence is also clear from the fingerprint plot delineated into Cl\u22efH/H\u22efCl , and Cl\u22efO/O\u22efCl contacts . The contribution from C\u22efH/H\u22efC contacts and other contacts (Table\u00a03The chlorine atom on the benzene (C14\u2013C19) ring makes a useful contribution to the mol\u00adecular packing. The small, ts Fig.\u00a06g to thems Fig.\u00a05c. Its pCl Fig.\u00a06d, and Cts Fig.\u00a06h. The cts Fig.\u00a06f and ots Table\u00a03, includiet al., 2016i.e. (II) as eluent to afford \u03b1-lapachone and \u03b2-lapachone (I)Referring to the reaction scheme, in a double-necked flask equipped with a magnetic bar and reflux condenser, under a nitro\u00adgen atmosphere, lawsone , paraformaldehyde , the vinyl selenide and the ionic liquid 1-butyl-3-methyl\u00adimidazolium chloride, [Bmim]Cl were added over 1,4-dioxane (2\u2005ml). The reaction mixture was heated at 383\u2005K and stirred over 2\u2005h. The reaction mixture was cooled and diluted with di\u00adchloro\u00admethane (100\u2005ml) and then washed with water (3 \u00d7 50\u2005ml). The organic phase was dried over NaUiso(H) set to 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989017007605/wm5392sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017007605/wm5392Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017007605/wm5392Isup3.cmlSupporting information file. DOI: 1551641CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The B\u2014H groups exhibit an intra\u00admolecular close contact with a C\u2014H group of the pyridine ring, which may be evidence of electrostatic attraction between the hydridic B\u2014H and the electropositive aromatic C\u2014H.The asymmetric unit contains two independent mol\u00adecules, which exhibit coplanar, mostly 14H19BN2O, contains two independent mol\u00adecules in the asymmetric unit. Both molecules exhibit coplanar, mostly sp2-hybridized meth\u00adoxy and di\u00admethyl\u00adamino substituents on their respective aromatic rings, consistent with \u03c0-donation into the aromatic systems. The B\u2014H groups exhibit an intra\u00admolecular close contact with a C\u2014H group of the pyridine ring, which may be evidence of electrostatic attraction between the hydridic B\u2014H and the electropositive aromatic C\u2014H. There appears to be weak C\u2014H\u22ef\u03c0(arene) inter\u00adactions between two of the H atoms of an amino\u00admethyl group and the meth\u00adoxy-substituted benzene ring of the other independent mol\u00adecule, and another C\u2014H\u22ef\u03c0 (arene) inter\u00adaction between one of the pyridine ring H atoms and the same benzene ring.The title compound [systematic name: 4-(di\u00admethyl\u00adamino)\u00adpyridine\u20134-meth\u00adoxy\u00adphenyl\u00adborane (1/1)], C RBH2) have been the focus of chemical research for over fifty years, most notably for their use in the indispensable hydro\u00adboration reaction, which permits reduction of olefins, carbonyl compounds and others and C1\u2032\u2014B1\u2032\u2014N1\u2032 = 111.0\u2005(1)\u00b0] . The B1\u2014C1 and B1\u2032\u2014C1\u2032 distances are consistent with a formal C\u2014B single bond. The oxygen atom of both meth\u00adoxy groups appears to be mostly 2sp hybridized, [C7\u2014O1\u2014C4 = 117.3\u2005(1) and C7\u2032\u2014O1\u2032\u2014C4\u2032 = 117.4\u2005(1)\u00b0] and is close to coplanar with the phenyl ring [torsion angles C7\u2014O1\u2014C4\u2014C3 = \u22127.4\u2005(2) and C7\u2032\u2014O1\u2032\u2014C4\u2032\u2014C3\u2032 = \u22127.1\u2005(2)\u00b0], consistent with \u03c0-donation into the phenyl ring.The asymmetric unit contains two independent mol\u00adecules Figs. 1 and 2 \u25b8 2sp hybridized [torsion angles C13\u2014N2\u2014C10 = 121.0\u2005(1)\u00b0 and C13\u2032\u2014N2\u2032\u2014C10\u2032 = 122.2\u2005(1)\u00b0] and is close to coplanar [torsion angles C13\u2014N2\u2014C10\u2014C11 = 2.4\u2005(2) and C13\u2032\u2014N2\u2032\u2014C10\u2032\u2014C11\u2032 = 3.4\u2005(1)\u00b0] consistent with \u03c0-donation into the pyridine ring.The geometries of the 4-(dimethylamino)pyridine (DMAP) fragment of both mol\u00adecules is similar to other structures of DMAP\u2013borane adducts. The nitro\u00adgen atom of the di\u00admethyl\u00adamino fragment appears to be Database survey). Inter\u00adestingly, the B\u2014H atoms exhibit intra\u00admolecular close contacts with the C\u2014H atoms of the pyridine ring [H12\u22efH2B = 2.26\u2005(3) and H12\u2032\u22efH2B\u2032 = 2.27\u2005(3)\u2005\u00c5] and are close to coplanar [torsion angles H2B\u2014B1\u2014N1\u2014C12 = 4(1) and H2B\u2014B1\u2014N1\u2014C12 = 16\u2005(1)\u00b0], which may be evidence of electrostatic inter\u00adactions between the hydridic B\u2014H atoms and electropositive aromatic C\u2014H atoms, and is observed in other DMAP\u2013borane adducts (see Database Survey). The planes of the pyridine rings and the benzene rings are almost normal to one another [the dihedral angle between the C1\u2013C6 and C8\u2013C12/N1 rings is 73.14\u2005(7)\u00b0 and that between the C1\u2032\u2013C6\u2032 and C8\u2032\u2013C12\u2032/N1\u2032 rings is 74.15\u2005(7)\u00b0]. Perhaps the most significant difference between the two mol\u00adecules is the 9.0\u00b0 difference in the torsion angle about the B\u2014N bond [C1\u2014B1\u2014N1\u2014C8 = \u221263.9\u2005(2) while C1\u2032\u2014B1\u2032\u2014N1\u2032\u2014C8\u2032 = \u221272.9\u2005(2)\u00b0] and 1.595\u2005(2)\u2005\u00c5, respectively] are consistent with formal N\u2014B single bonds, and are within the range observed for other DMAP\u2013borane adducts inter\u00adactions between two of the hydrogen atoms of the amino\u00admethyl group and the meth\u00adoxy\u00adphenyl group of a neighboring mol\u00adecule inter\u00adactions described in the previous section yielded four structures: UTOZEJ , followed by extraction with anhydrous di\u00adchloro\u00admethane (4\u2005mL). The extract was filtered through a 0.45\u2005\u00b5m PTFE syringe filter. The solvent was again removed in vacuo to afford a white solid . Crystals suitable for X-ray diffraction were grown by diffusion of pentane into a concentrated solution of the title compound in anhydrous di\u00adchloro\u00admethane.In a nitro\u00adgen-filled glove box, sodium 4-meth\u00adoxy\u00adphenyl\u00adborohydride and 4-di\u00admethyl\u00adamino\u00adpyridine were combined in a 20\u2005mL vial containing a stir bar and dissolved in anhydrous THF (4\u2005mL). The solution was cooled to 247\u2005K in the freezer and chloro\u00adtri\u00admethyl\u00adsilane was added dropwise 1H NMR \u03b4 (ppm): 8.12 , 7.23 , 6.80 , 6.52 , 3.78 , 3.11 . 13C NMR \u03b4 (ppm): 157.3, 154.9, 146.7, 145.0 (br s), 134.5, 122.9, 106.5, 55.0, 39.5. 11B NMR \u03b4 (ppm): \u22125.0 . FTIR : 3012, 2952, 2923, 2853, 2610, 2346, 2288, 2227, 1634, 1548, 1442, 1418, 1392, 1237, 1223, 1161, 1076, 1031, 811, 797, 548, 515.Uiso(H) = 1.5Ueq(C-methyl) and 1.2eq(C) for other H atoms. The B-bound H atoms were located in a difference-Fourier map and freely refined. Methyl H atoms were refined without restrictions on rotation around the C\u2014C bonds, HFIX 138 in SHELXL I. DOI: 10.1107/S2056989017015171/lh5853Isup2.hklStructure factors: contains datablock(s) I. DOI: 1580559CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The configuration about each double bond in the N\u2014N=C\u2014C=C chain is E; the chain has an all trans conformation. In the crystal, N\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions link the mol\u00adecules into a three-dimensional network.The title di\u00adthio\u00adcarbazate ester has approximate mirror symmetry with the putative plane bis\u00adecting the \u2013CH 18H18N2S2 [systematic name: (E)-4-methyl\u00adbenzyl 2-[(E)-3-phenyl\u00adallyl\u00adidene]hydrazinecarbodi\u00adthio\u00adate, comprises an almost planar central CN2S2 residue [r.m.s. deviation = 0.0131\u2005\u00c5]. The methyl\u00adene(tolyl-4) group forms a dihedral angle of 72.25\u2005(4)\u00b0 with the best plane through the remaining non-hydrogen atoms [r.m.s. deviation = 0.0586\u2005\u00c5] so the mol\u00adecule approximates mirror symmetry with the 4-tolyl group bis\u00adected by the plane. The configuration about both double bonds in the N\u2014N=C\u2014C=C chain is E; the chain has an all trans conformation. In the crystal, eight-membered centrosymmetric thio\u00adamide synthons, {\u22efHNCS}2, are formed via N\u2014H\u22efS(thione) hydrogen bonds. Connections between the dimers via C\u2014H\u22ef\u03c0 inter\u00adactions lead to a three-dimensional architecture. A Hirshfeld surface analysis shows that (I) possesses an inter\u00adaction profile similar to that of a closely related analogue with an S-bound benzyl substituent, (II). Computational chemistry indicates the dimeric species of (II) connected via N\u2014H\u22efS hydrogen bonds is about 0.94 kcal mol\u22121 more stable than that in (I).The title di\u00adthio\u00adcarbazate ester (I), C The dihedral angles between the central group and the S2- and N2-bound substituents are 71.65\u2005(4) and 7.08\u2005(8)\u00b0, respectively. The dihedral angle between the outer groups is 72.33\u2005(4)\u00b0 and is indicative of an approximately orthogonal relationship. Indeed, the r.m.s. deviation of all non-hydrogen atom in (I)2(tolyl-4) residue is 0.0586\u2005\u00c5, and the angle between this plane and that through the CH2(tolyl-4) residue is 72.25\u2005(4)\u00b0. The 1,4-carbon atoms of the 4-tolyl ring lie on the approximate mirror plane defined by the rest of the mol\u00adecule with the remaining pairs of ring atoms being related across the putative plane.The mol\u00adecular structure of (I)E in each case. This implies the N1\u2014N2=C2\u2014C3=C4 sequence has an all trans conformation as seen in the N1\u2014N2\u2014C2\u2014C3, N2\u2014C2\u2014C3\u2014C4 and C2\u2014C3\u2014C4\u2014C5 torsion angles of 177.41\u2005(13), \u2212178.70\u2005(15) and 178.23\u2005(15)\u00b0, respectively. The C1\u2014S2 [1.7455\u2005(16)\u2005\u00c5] and, especially, C11\u2014S2 [1.8233\u2005(16)\u2005\u00c5] bond lengths are considerably longer than the C1\u2014S1 bond [1.6752\u2005(16)\u2005\u00c5] consistent with considerable thione character in the latter. This is borne out also by the observation that the angles about the C1 atom involving S1 are wider, by over 7\u00b0, i.e. S1\u2014C1\u2014S2 = 125.20\u2005(10)\u00b0 and N1\u2014C1\u2014S1 121.06\u2005(12)\u00b0, cf. N1\u2014C1\u2014S2 of 113.74\u2005(11)\u00b0.The configuration about the C2=N2 imine [1.284\u2005(2)\u2005\u00c5] and C3=C4 ethene [1.339\u2005(2)\u2005\u00c5] bonds is Computational chemistry calculations.Further discussion on the mol\u00adecular geometry of (I)2 mediated by N\u2014H\u22efS(thione) hydrogen bonds, Fig.\u00a02a and Table\u00a01b, via methyl\u00adene-C\u2014H\u22ef\u03c0(tol\u00adyl), tolyl-C\u2014H\u22ef\u03c0(phen\u00adyl) and phenyl-C\u2014H\u22ef\u03c0(tol\u00adyl) inter\u00adactions, Table\u00a01bc plane and these are linked by the N\u2014H\u22efS hydrogen bonds.The most prominent feature of the mol\u00adecular packing is the formation of an eight-membered, centrosymmetric thio\u00adamide synthon, {\u22efHNCS}et al., 20082(tolyl-4) group, that might be regarded as the \u2018parent\u2019 compound, hereafter referred to as (II). While detailed discussion on the comparison of their mol\u00adecular geometries and computational modelling are given in Computational chemistry calculations, the present section focuses upon the study of inter\u00admolecular inter\u00adactions formed by (I)et al., 2016The most closely related compound in the crystallographic literature is one with a benzyl substituent at the S2 atom (Tarafder ca 25.4% for (I)ca 17.5 and 16.9%], N\u22efH/H\u22efN [ca 5.6 and 5.5%] as well as other minor inter\u00adactions including N\u22efC/C\u22efN, S\u22efC/C\u22efS and S\u22efN/N\u22efS, which constitute less than 5% of the overall contacts.Both (I)divs de at the inter\u00advals of 0.01\u2005\u00c5 reveals that (I)a,. Specifically, the decomposed fingerprint plot of H\u22efH for (I)de + di contact distance of 1.96\u2005\u00c5 which is approximately 0.43\u2005\u00c5 (17%) shorter cf. 2.36\u2005\u00c5 for (II), Fig.\u00a04b. Both (I)c, at approximately 2.7\u2005\u00c5, which is slightly shorter than the van der Waals radii of 2.9\u2005\u00c5. The decomposed fingerprint plots of S\u22efH/H\u22efS and N\u22efH/H\u22efN contacts for (I)cf. (II), in which the difference is merely 0.04\u2005\u00c5 (1.7%). Similarly, the S\u22efH/ H\u22efS contacts of both (I)cf. the sum of their van der Waals radii by 0.53 and 0.58\u2005\u00c5, respectively (21.5 and 24.0%). As a result, those contacts display intense red spots on their Hirshfeld surface, Fig.\u00a04d.A detailed comparison of the two-dimensional fingerprint plots of \u22efS Fig.\u00a04d and N\u22efts Fig.\u00a04e for , exhibits a greater mol\u00adecular volume and surface area, and is slightly less globular. This results in a lower surface-to-volume ratio and density for (I)In view of the close structural similarity between (I)et al., 2016As mentioned in the previous section, the \u2018parent\u2019 compound represents the most closely related analogue to (I)GaussView5 and pre-optimized using a semi empirical method (PM6) with a precise self-consistent field criterion. Subsequently, the geometries were further optimized at B3LYP/6-311+G without imposing symmetry constraints. A frequency analysis was performed on each optimized structure using the same level of theory and basis set to validate that each structure was indeed the local minimum structure with no imaginary frequency. All calculations were performed using the Gaussian09 software package ca 0.02 and 0.03\u2005\u00c5, respectively. In the chain, the C1\u2014N1 bond lengths have lengthened by ca 0.03\u2005\u00c5, a difference accompanied by a contraction in the N1\u2014N2 bond length by about the same amount. Minor differences are also noted in bond angles with widening of S1\u2014C1\u2014S2 and the angles subtended at the nitro\u00adgen atoms by 2\u20133\u00b0 with similar contractions in the C1\u2014S1\u2014C11 and S1\u2014C1\u2014N1 angles.The results, as shown from the superposition of the experimental structure and theoretical model of (I)d,p) basis set. It has been demonstrated that the long-range corrected hybrid method can greatly reduce self-inter\u00adaction errors was obtained upon the correction of basis set superposition error (BSSE) by counterpoise correction. All calculations were performed in gas phase using Gaussian09 software EBSSEint) of \u221212.92 and \u221213.86 kcal mol\u22121, respectively. The range is approximately 3.89 to 5.23 kcal mol\u22121 less than the energy computed for a pair of thio\u00adurea dimers at the RIMP2/cc-pVDZ and cc-pVTZ levels of theory is lower in energy cf. (I)cf. (I)The dimeric species of (I)et al., 2010S-4-methyl\u00adbenzyl\u00addithio\u00adcarbazate was dissolved in hot aceto\u00adnitrile (100\u2005ml) and added to an equimolar amount of cinnamaldehyde in absolute ethanol (20\u2005ml). The mixture was heated for about 2\u2005h and was then allowed to stand overnight. The pale-brown crystals that formed were filtered and washed with absolute ethanol at room temperature. Yield: 70%. M.p. 463\u2013466\u2005K. Analysis: Calculated for C18H18N2S2: C, 66.22; H, 5.56; N, 8.58. Found: C, 65.87; H, 5.77; N, 9.00%. FT\u2013IR : 3102, \u03bd(N\u2014H); 1613, \u03bd(C=N); 1021, \u03bd(N\u2014N); 749, \u03bd(CSS).The following procedure was adapted from the literature (Ravoof Uiso(H) set to 1.2\u20131.5Ueq(C). The nitro\u00adgen-bound H atom was located in a difference-Fourier map but was refined with a distance restraint of N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989017003991/hb7666sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017003991/hb7666Isup2.hklStructure factors: contains datablock(s) I. DOI: 1537500CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III defined by an N4O3 donor set. The packing features supra\u00admolecular layers sustained by C\u2014H\u22efO, C\u2014H\u22ef\u03c0(ar\u00adyl) and C\u2014Cl\u22ef\u03c0(ar\u00adyl) inter\u00adactions.The title compound features an amine-N-capped octa\u00adhedral coordination geometry for Yb III atom in the title complex, [Yb(C27H24Cl3N4O3)] }tris\u00ad(4-chloro\u00adphenolato)ytterbium(III)], is coordinated by a trinegative, hepta\u00addentate ligand and exists within an N4O3 donor set, which defines a capped octa\u00adhedral geometry whereby the amine N atom caps the triangular face defined by the three imine N atoms. The packing features supra\u00admolecular layers that stack along the a axis, sustained by a combination of aryl-C\u2014H\u22efO, imine-C\u2014H\u22efO, methyl\u00adene-C\u2014H\u22ef\u03c0(ar\u00adyl) and end-on C\u2014Cl\u22ef\u03c0(ar\u00adyl) inter\u00adactions. A Hirshfeld surface analysis points to the major contributions of C\u22efH/ H\u22efC and Cl\u22efH/H\u22efCl inter\u00adactions to the overall surface but the Cl\u22efH contacts are at distances greater than the sum of their van der Waals radii.The Yb Being large and having seven potential donor atoms, these ligands are capable of coordinating lanthanides even if the atomic sizes of lanthanides are greater in comparison to their transition metal counterparts Despite being less studied than transition metal complexes, the crystal chemistry of lanthanides is rich and diverse as their complexes can display various coordination numbers and geometries that are not readily predicted \u00b0, indicating a parallel disposition. There is a range in the Yb\u2014O bond lengths, i.e. >0.02\u2005\u00c5, with the shortest Yb\u2014O1 bond being trans to the most loosely bound imine-N4 atom, and the longest Yb\u2014O2 bond being trans to most tightly held imine-N2 atom, Table\u00a01The mol\u00adecular structure of (I)b axis, Fig.\u00a02a. These are reinforced by methyl\u00adene-C\u2014H\u22ef\u03c0(ar\u00adyl) contacts, also shown in Fig.\u00a02a. Chains are connected into supra\u00admolecular layers in the bc plane by end-on C\u2014Cl\u22ef\u03c0(ar\u00adyl) inter\u00adactions, Fig.\u00a02b. Layers stack along the a axis with no directional inter\u00adactions between them, Fig.\u00a02c dnorm contact distances within the range of \u22120.18 to 1.65\u2005\u00c5 through calculation of the inter\u00adnal (di) and external (de) distances of a particular Hirshfeld surface point to its nearest nucleus a shows a butterfish-like two-dimensional fingerprint plot for (I)ca 26 and 5%, respectively, of the overall inter\u00adactions in the crystal, with de + di distances of \u223c2.54\u2005\u00c5 and \u223c2.43\u2005\u00c5, respectively, Fig.\u00a03b and 3c. As seen from the images of Fig.\u00a04d. Finally, the Cl\u22efH/H\u22efCl inter\u00adactions appear as the third most dominant inter\u00adaction, right after H\u22efH and C\u22efH/H\u22efC, with an overall contribution of ca 24% to the Hirshfeld surface, despite the fact that these inter\u00adactions are considered weak with contact distances greater than the sum of van der Waals radii. However, as seen from Fig.\u00a03e, these inter\u00adactions are responsible for the appearance of the tails of the \u2018butterfish-shape\u2019.The Hirshfeld surface analysis was performed on (I)III complex has been the subject of two independent determinations structures are available for comparison where the ligand is identical to that in (I)i.e. P21/n, for Ln = Sm bond lengths. The only anomalous parameter might be the length of the Gd\u2014N(amine) bond, i.e. 2.737\u2005(8)\u2005\u00c5, the relatively high standard uncertainty value notwithstanding.A wider range of lanthanide : 1628 (s) \u03bd(C=N), 1517 (m), 1449 (m), 1392 (m) \u03bd(\u2013O\u2014C=C\u2013), 1158 (m) \u03bd(C\u2014O\u2014C). Analysis calculated for C27H24Cl3N4O3Yb: C, 44.31; H, 3.31; N, 7.66%. Found: C, 44.63; H, 3.12; N, 7.92%.The Schiff base ligand, tris\u00ad{[amino]\u00adeth\u00adyl}amine (Kanesato Uiso(H) set to 1.2Ueq(C). Owing to poor agreement, one reflection, i.e. (7 0 4), was omitted from the final cycles of refinement. The maximum and minimum residual electron density peaks of 1.65 and 0.45\u2005e\u2005\u00c5\u22123, respectively, were located 0.84 and 1.51\u2005\u00c5 from the Yb and N2 atoms, respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989016013748/hb7612sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016013748/hb7612Isup2.hklStructure factors: contains datablock(s) I. DOI: 1501230CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "O:O\u2032-bridged dinuclear unit which is extended through N-atom donors of the pyrimidine ligand into a two-dimensional layered structureThe coordination polymeric silver(I)\u2013diclofenac complex including pyrimidine is based on a centrosymmetric carboxyl\u00adate 14H11Cl2NO2) (diclH) and pyrimidine (pym), namely poly[{\u03bc2-2-[2-phen\u00adyl]acetato-\u03ba2O:O\u2032}(\u03bc2-pyrimidine-\u03ba2N1:N3)silver(I)], [Ag(C14H10Cl2NO2)(C4H4N2)]n or [Ag(\u03bc-dicl)(\u03bc-pym)]n, the very distorted tetra\u00adhedral AgN2O2 coordination centres comprise two N-atom donors from bridging pym ligands [Ag\u2014N = 2.381\u2005(3) and 2.412\u2005(3)\u2005\u00c5] and two carboxyl\u00adate O-atom donors from dicl ligands [Ag\u2014O = 2.279\u2005(2) and 2.280\u2005(2)\u2005\u00c5], which bridge Ag atoms, giving a centrosymmetric dinuclear units with a short Ag\u22efAg separation [2.8931\u2005(5)\u2005\u00c5]. Within the units are short intra\u00adligand C\u2014Cl\u22ef\u03c0(pym) inter\u00adactions [3.6409\u2005(15)\u2005\u00c5]. The units are linked through the bridging N atoms of the pym ligand into a two-dimensional sheet\u2013polymer structure lying parallel to (100) and stabilized by inter-ring \u03c0\u2013\u03c0 inter\u00adactions between the pym ligands [Cg\u22efCg = 3.4199\u2005(17)\u2005\u00c5]. Additional inter-unit C\u2014H\u22efO and C\u2014H\u22efCg hydrogen-bonding inter\u00adactions between the sheets give an overall three-dimensional structure.In the title mixed-ligand silver(I) coordination polymeric complex with the non-steroidal anti-inflammatory drug diclofenac (C These studies have shown that short Ag\u22efAg separations are one of the most important factors for the manifestation of such properties from separate dicl ligands and two nitro\u00adgen atoms /141\u00b0}, where \u03b1 and \u03b2 are the largest angles around the metal atom) is 0.732 and indicates substantial deviation from ideal tetra\u00adhedral geometry 2(\u03bc-pym)2]n n 2] units \u2005\u00c5] is significantly shorter than the sum of the van der Waals radii for two silver atoms (3.44\u2005\u00c5), indicating weak inter\u00adactions between adjacent AgI ions, forming an [Ag2(COO)2] units. If coexisting strong argentophilic Ag1\u22efAg1i inter\u00adactions are considered as coordinative, it could be reasoned that the coordination around Ag1 is slightly distorted trigonal\u2013bipyramidal [the structural distortion index tau (\u03c4) was calculated to be 0.06] ds Fig.\u00a01. The dists Fig.\u00a02. Within 2-1N,N-bridging ligand between neighboring [Ag2(COO)2] units, leading to the formation of a two-dimensional coordination polymer, extending along (100) 2] units, which comprise eight-membered rings, can be defined as the nodes of the structure. Connection of the four different pym ligands to these nodes provides continuity of the structure Fig.\u00a04. In othere Fig.\u00a04.carbox\u00adyl hydrogen-bonding inter\u00adaction [2.971\u2005(3)\u2005\u00c5] \u00b0, the conformation of the ligand being stabilized by an intra\u00admolecular N1\u2014H1\u22efO2] Table\u00a02.iii hydrogen-bonding inter\u00adaction stabilizes the crystal packing (Table\u00a02Cg6iv inter\u00adaction to a pym ring [3.983\u2005\u00c5] and a strong \u03c0\u2013\u03c0 stacking inter\u00adaction between aromatic rings of the pym ligands , shown in Fig.\u00a03In the crystal, a C16\u2014H16\u22efO1g Table\u00a02. In addi3) of AgNO3 with stirring. A white suspension with a white precipitate formed and the addition of aceto\u00adnitrile (10\u2005cm3) to this resulted in a clear solution which was left to stand for slow evaporation in darkness at room temperature. Single crystals of (I)All reactions were performed with commercially available reagents and used without further purification. Solid sodium 2-phenyl\u00adacetate (Nadicl) and pyrimidine were added to an aqueous solution in the frequency range 4000\u2013600\u2005cmas) and symmetric (\u03c5s) vibrations of the carboxyl\u00adate group. The difference between the asymmetric and symmetric carboxyl\u00adate stretching [\u0394\u03bd = \u03c5as(COO\u2212) - \u03c5s(COO\u2212)] is often used to correlate the infrared spectra of metal carboxyl\u00adate structures. When \u0394\u03bd < 200\u2005cm\u22121, the carboxyl\u00adate groups of the complexes can be considered bidentate = 1.2Ueq(C). The N-bound H atom was located in a difference-Fourier map but was also allowed to ride in the refinement with Uiso(H) = 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989016014730/zs2370sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016014730/zs2370Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016014730/zs2370sup3.tifSupporting information file. DOI: 1500646CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports7: Article number: 45206; 10.1038/srep45206 published online: 05302017; updated: 12222017.This Article contains errors. In the legend of Figure 1,a) Spectral flux per bunch through an on-axis 1\u2009mm2 area delivered by Beamline ID19 in the four bunch mode (40\u2009mA storage ring current), including 2.8\u2009mm diamond and 1.4\u2009mm aluminium filtering. The total flux was 1.2\u2009\u00d7\u2009109 photons s\u22121 mm\u22122 on-axis.\u201d\u201c(should read:a) Spectral flux per bunch through an on-axis 1\u2009mm2 area delivered by Beamline ID19 in the four bunch mode (40\u2009mA storage ring current), including 2.8\u2009mm diamond and 1.4\u2009mm aluminium filtering. The total flux was 4\u2009\u00d7\u2009107 photons/bunch/mm2 on-axis.\u201d\u201c delivered per bunch (150\u2009ps duration23) on Beamline ID19 in the four bunch mode.\u201d\u201cFigure 1(a) shows the calculated on-axis spectral X-ray flux (1.2\u2009\u00d7\u200910should read:7 photons/bunch/mm2 on-axis) delivered per bunch (150\u2009ps duration23) on Beamline ID19 in the four bunch mode.\u201d\u201cFigure 1(a) shows the calculated on-axis spectral X-ray flux (4\u2009\u00d7\u200910In the Discussion section,\u201cThe time at which the leading lateral release waves reached the shock front on-axis in the PC-PC and Cu-PC loadings were estimated to be 3.30\u2009\u03bcs and 2.53\u2009\u03bcs, respectively.\u201dshould read:\u201cThe times at which the leading lateral release waves reached the shock front on-axis in the PC-PC and Cu-PC loadings were estimated to be 2.3 \u03bcs and 1.9 \u03bcs, respectively.\u201d"} +{"text": "Scientific Reports7: Article number: 42192; 10.1038/srep42192 published online: 02082017; updated: 01292018.This Article contains an error in Figure 2. For CYP3A5 *3\u2009=\u2009AG without hypertension,\u201cN\u2009=\u200931 Dose\u2009=\u20093.73\u201dshould read:\u201cN\u2009=\u200931 Dose\u2009=\u20094.13\u201dThe correct Figure 2 appears below as"} +{"text": "The packing features C\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions.With both chelating and \u03bc 2(C8H8NS2)4], is a centrosymmetric dimer with both chelating and \u03bc2-tridentate di\u00adthio\u00adcarbamate ligands. The resulting S5 donor set defines a CdII coordination geometry inter\u00admediate between square-pyramidal and trigonal\u2013bipyramidal, but tending towards the former. The packing features C\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions, which generate a three-dimensional network. The influence of these inter\u00adactions, along with intra-dimer \u03c0\u2013\u03c0 inter\u00adactions between chelate rings, has been investigated by an analysis of the Hirshfeld surface.The title compound, [Cd The S3 di\u00adthio\u00adcarbamate ligand is strictly chelating, with \u0394(Cd\u2014S) = 0.08\u2005\u00c5. Reflecting the symmetric modes of coordination of the di\u00adthio\u00adcarbamate ligands, the C\u2014S bond lengths are equal within 5\u03c3 (Table\u00a01The centrosymmetric binuclear mol\u00adecule of (I)\u03c3 Table\u00a01.5 donor set defines a highly distorted penta\u00adcoordinate geometry, with the major distortions due to the disparate Cd\u2014S bond lengths and the acute angles subtended at the CdII atom by the chelating ligands \u00b0. A measure of the distortion of a coordination geometry from the ideal square-pyramidal and trigonal\u2013bipyramidal geometries is given by the value of \u03c4 or Fig.\u00a02.et al., 2016dnorm in the range \u22120.055 to 1.371 au and 5(b), respectively, and again highlight the influence of C\u2014H\u22efS inter\u00adactions, short C10\u22efC15 contacts and C\u2014H\u22ef\u03c0 inter\u00adactions involving phenyl rings (atoms C3\u2013C8) as the acceptor. Thus, the C\u2014H\u22efS inter\u00adactions involving the phenyl-ring C4, C5 and H5 atoms with S1 are shown with black dashed lines in Fig.\u00a05a); the red dashed lines indicate short inter\u00adatomic C\u22efC contacts .The Hirshfeld surface analysis for (I)au Fig.\u00a03, the briau Fig.\u00a03. The donau Fig.\u00a03 give risau Fig.\u00a03. The imms Table\u00a03. The C\u2014Het al., 2007a)\u2013(e); their relative contributions to the Hirshfeld surface are summarized qu\u00adanti\u00adtatively in Table\u00a04b) that H\u22efH contacts do not exert much influence on the mol\u00adecular packing, as their inter\u00adatomic distances are greater than the sum of their van der Waals radii, i.e. de + di > 2.8\u2005\u00c5. A pair of peaks appearing in the fingerprint plot delineated into S\u22efH/H\u22efS contacts at de + di \u223c 2.8\u2005\u00c5 arise from the C5\u2014H5\u22efS1 inter\u00adaction; the weaker C4\u22efH4\u22efS1 inter\u00adaction and short inter\u00adatomic H\u22efS/S\u22efH contacts involving the S3 atom indicate the significance of these contacts through the presence of C\u2014H\u22ef\u03c0 inter\u00adactions and short inter\u00adatomic C\u22efH/H\u22efC contacts in the crystal. A pair of green lines within the forceps also indicates the influence of these contacts. Finally, an arrow-shaped distribution of green points in the centre in the plot corresponding to S\u22efS contacts , together with the contribution from Cd\u22efS/S\u22efCd contacts to the Hirshfeld surface (Table\u00a04Cg\u22efCg = 3.6117\u2005(11)\u2005\u00c5; symmetry code: \u2212x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z]. The small contributions from Cd\u22efH/H\u22efCd and N\u22efH/H\u22efN contacts \u22121. 1H and 13C NMR spectra were recorded at room temperature in DMSO-d6 solution on a Jeol ECA 400\u2005MHz FT\u2013NMR spectrometer.All chemicals and solvents were used as purchased without purification, and all reactions were carried out under ambient conditions. The melting point was determined using an Electrothermal digital melting-point apparatus and was uncorrected. The IR spectrum was obtained on a PerkinElmer Spectrum 400 FT Mid-IR/Far-IR spectrophotometer from 4000 to 400\u2005cm\u22121): 1491 (m) [\u03bd(C\u2014N)], 1160 (m), 964 (s) [\u03bd(C\u2014S)] cm\u22121. 1H NMR: \u03b4 7.26\u20137.42 , 2.05 . 13C NMR: \u03b4 46.6 (Me) 125.6, 128.4, 129.6, 147.9 (aromatic C), 207.8 (CS2).Sodium methyl\u00adphenyl\u00addithio\u00adcarbamate in methanol (25\u2005ml) was added to cadmium chloride in methanol (10\u2005ml). The resulting mixture was stirred and refluxed for 2\u2005h. The filtrate was evaporated until an off-white precipitate was obtained, which was recrystallized in methanol. Slow evaporation of the filtrate yielded colourless crystals of the title compound . IR (cmUiso(H) values set at 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989017002705/hb7659sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017002705/hb7659Isup2.hklStructure factors: contains datablock(s) I. DOI: 1533246CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Prunus domestica ssp. domestica), six colorless non\u2010fluorescent chlorophyll catabolites (NCCs) were characterized, named Pd\u2010NCCs. In addition, several minor NCC fractions were tentatively classified. The structure of the most polar one of the NCCs, named Pd\u2010NCC\u201032, featured an unprecedented twofold glycosidation pattern. Three of the NCCs are also functionalized at their 32\u2010position by a glucopyranosyl group. In addition, two of these glycosidated NCCs carry a dihydroxyethyl group at their 18\u2010position. In the polar Pd\u2010NCC\u201032, the latter group is further glycosidated at the terminal 182\u2010position. Four other major Pd\u2010NCCs and one minor Pd\u2010NCC were identified with five NCCs from higher plants known to belong to the \u2018epi\u2019\u2010series. In addition, tentative structures were derived for two minor fractions, classified as yellow chlorophyll catabolites, which represented oxidation products of two of the observed Pd\u2010NCCs. The chlorophyll catabolites in leaves of plum feature the same basic structural pattern as those found in leaves of apple and pear trees.In cold extracts of senescent leaves of the plum tree ( Hv\u2010NCC\u20101 from senescent leaves of barley (Hordeum vulgare).Hv\u2010NCC\u20101 as a 1\u2010formyl\u201019\u2010oxobilin\u2010type linear tetrapyrrolepFCCs), with species\u2010dependent configuration of their formation.pFCCs are rapidly hydroxylated to 32\u2010OH\u2010pFCC (probably still in the chloroplast).Fig.\u00a0About 25\u00a0years ago, chlorophyll (Chl) breakdown and the appearance of the fall colors were still a stunning mystery.Table\u00a0In the meantime, NCCs have been found in extracts of senescent leaves of a range of plants,Fig.\u00a0However, as was recognized recently, Chl\u2010breakdown \u2018branches out\u2019, and furnishes \u20181,19\u2010dioxobilin\u2010type\u2019 Chl\u2010catabolites (DCCs)Prunus domestica ssp. domestica). As shown below, Chl\u2010breakdown in senescent leaves of this fruit tree follows the \u2018PaO/phyllobilin\u2019 pathway of Chl\u2010breakdown.epi\u2010type\u2019. In addition, in the extracts several YCCs were also found.In the context of investigations of Chl\u2010catabolites in domestic agricultural plants, we have studied the nature of such phyllobilins in stone fruit and report here our work on the Chl\u2010catabolites in leaves of the plum tree (Prunus domestica) and frozen for storage. Five major and nine minor colorless NCCs were provisionally identified in extracts of senescent leaves of plum trees on the basis of their characteristic UV\u2010absorbance properties, using analytical HPLC , as first reported in the spectrum of Hv\u2010NCC\u20101.Yellow senescent and green leaves were collected from plum trees , analyzed by UV/VIS\u2010spectroscopy first and of Hv\u2010NCC\u20101R)\u2010configuration at the stereogenic C(10).For spectroscopic analysis of the most abundant NCCs in the leaves of Pd\u2010NCC\u201032 (1) displayed a strong signal at m/z 1025.3839, corresponding to C47H62N4NaO20+ and establishing the molecular formula as C47H62N4O20. Likewise, a positive\u2010ion\u2010mode ESI\u2010MS spectrumFig.\u00a0pseudo\u2010molecular ion [M\u00a0+\u00a0H]+ at m/z 1003.1, also consistent with the molecular formula of C47H62N4O20. Characteristic fragment ions at m/z 841.2, 684.1 and 679.2 were also detected, which indicated the loss of a sugar moiety (C6H10O5) from [M\u00a0+\u00a0H]+,D in CD3OD at 10\u00a0\u00b0C ) at low field, four Me group singlets at high field and a singlet for the methyl ester group at 3.75\u00a0ppm. The typical signals for a peripheral vinyl group were not observed. From 1H,13C\u2010heteronuclear (HSQC and HMBC) and 1H,1H\u2010homonuclear NMR\u2010correlations (COSY and ROESY) of Pd\u2010NCC\u201032 (1) in CD3OD, assignment of the signals of 47 H\u2010atoms and 45 13C\u2010nuclei could be achieved (4.17\u00a0ppm) and H\u2013C(1\u2033) (4.33\u00a0ppm), as well as H\u2013C(2\u2032) (3.17\u00a0ppm) and H\u2013C(2\u2033) (3.21\u00a0ppm), the chemical shifts of the pairs of signals differed significantly . Chemical shifts and doublet nature (J\u00a0=\u00a07.8\u00a0Hz) of H\u2013C(1\u2032) and H\u2013C(1\u2033) indicated \u03b2\u2010anomeric attachment of both sugar moieties, as observed earlier for the 32\u2010glucopyranoside moieties of NCCs.1H\u2010 and 13C\u2010chemical shifts of Pd\u2010NCC\u201032 (1) with those of the known NCCs with a peripheral glucopyranosyl group at C(32).1H,13C\u2010HMBCs from H\u2013C(1\u2032) with C(32) and from H\u2013C(1\u2033) with C(182) established the attachment of one sugar moiety at each one of the terminal C\u2010atoms of the Et side chain at C(3) (ring A) and of the 1,2\u2010dihydroxyethyl group at C(18) (ring D). The 1H\u2010 and 13C\u2010chemical shifts at the positions C(182) and C(32) were also consistent with an attached peripheral sugar substituent. However, as with other 1,2\u2010dihydroxyethyl substituted NCCs,1 the configuration at C(182) remains unknown.In a 600\u00a0MHz Pd\u2010NCC fractions , Pd\u2010NCC\u201040 (3), Pd\u2010NCC\u201056 (4), Pd\u2010NCC\u201060 (5), and Pd\u2010NCC\u201071 (6) were also isolated and purified by HPLC. A positive\u2010ion\u2010mode ESI\u2010MS spectrum of Pd\u2010NCC\u201060 (5) showed a pseudo\u2010molecular ion [M\u00a0+\u00a0H]+ at m/z 645.2, consistent with the molecular formula of C35H40N4O8. Characteristic fragment ion peaks were visible at m/z 613.2 and 522.1, corresponding to the loss of MeOH and the loss of ring D from [M\u00a0+\u00a0H]+. The same molecular formula and fragmentation is known for the major NCC from Cercidiphyllum japonicum (Cj\u2010NCC\u20101),epi\u2019\u2010configuration at C(16).Pd\u2010NCC\u201060 (5) and Cj\u2010NCC\u20101 were separately analyzed by analytical HPLC, as well as a 1:1 mixture of both in a co\u2010injection of Pd\u2010NCC\u201060 (5). Consistent with their origin from a common \u2018primary\u2019 FCC,epi\u2010pFCC, the other colorless Pd\u2010NCCs were also deduced to belong to the epi\u2010series.Five other tion see , Fig.\u00a010Pd\u2010NCC\u201056 (4) was determined as C41H50N4O13 by ESI mass spectrometry, which furnished a base peak [M\u00a0+\u00a0H]+ at m/z 807.2. Fragment\u2010ions at m/z 775.3, 684.2 and 645.2 corresponded to the loss, alternatively, of MeOH, of ring D and of a hexose moiety from [M\u00a0+\u00a0H]+. These data indicate the presence of one hexopyranose moiety at the HO\u2013C(32) group of ring A of Pd\u2010NCC\u201056 (4) and a vinyl group at C(18) of ring D. This indicates a common chemical constitution of 4 and of Nr\u2010NCC\u20102Fig.\u00a0The molecular formula of Pd\u2010NCC\u201040 (3) could be deduced tentatively as C35H42N4O10 by ESI mass spectrometry, which showed the experimental base peak [M\u00a0+\u00a0H]+ at m/z 679.2. In the mass spectra, characteristic fragment\u2010ion peaks at m/z 647.2 and 522.1 were also detected, which corresponded to the loss of MeOH and to the loss of ring D (from [M\u00a0+\u00a0H]+). Accordingly, the catabolite Pd\u2010NCC\u201040 (3) indicated a pseudo\u2010molecular ion at m/z 841.2, consistent with the molecular formula of C41H52N4O15. The fragments at m/z 809.3, 684.2, 679.2 and 522.1 indicated the loss of MeOH, the loss of ring D, the loss of a sugar moiety and the loss of ring D and a sugar moiety. Thus, the catabolite 2 carries a sugar substituent at the C(3) hydroxyethyl side chain (ring A) and a 1,2\u2010dihydroxyethyl group at C(18) (ring D). According to their fragmentation pattern,Pd\u2010NCC\u201035 (2) was determined as C35H40N4O7 with a pseudo\u2010molecular ion at m/z 629.2. Fragments at m/z 597.2 and 506 indicate the loss of MeOH and ring D. Pseudo\u2010molecular ion and fragment\u2010ions are consistent with a chemical constitution of 6, as previously found for Cj\u2010NCC\u20102 and of Cj\u2010NCC\u20102 was supported by a common retention time of 6 and Cj\u2010NCC\u20102 in a HPLC co\u2010injection experiment.The molecular formula of Pd\u2010NCC\u201054) by LC/ESI\u2010MS revealed a pseudo\u2010molecular ion at m/z 661.2 ([M\u00a0+\u00a0H]+), consistent with the molecular formula of C35H40N4O9. We suspected Pd\u2010NCC\u201054 as product of the formal addition of an O\u2010atom to Pd\u2010NCC\u201060 (5) from an endogenous oxidation process. Indeed, as shown recently,2O may eliminate easily, resulting in corresponding YCCs.Pd\u2010YCC\u201067, which showed mass spectral data (pseudo\u2010molecular ion with m/z 643.2) consistent with its formation as the formal product of an oxidative dehydrogenation of Pd\u2010NCC\u201060 (5). A further minor fraction, classified as YCC from a prominent absorption maximum near 420\u00a0nm, was also subjected further to ESI\u2010MS analysis. The latter data suggested Pd\u2010YCC\u201061 (m/z 805.1) to represent a YCC derived from oxidation of the glucosylated Pd\u2010NCC\u201056 (4). When extracts were prepared after storage of senescent leaves of the plum tree at room temperature for 7\u00a0min, an increase of the content of both YCCs (Pd\u2010YCC\u201061 and Pd\u2010YCC\u201067) was observed, as well as the formation of 15\u2010OH\u2010Pd\u2010NCC\u201060, identified by comparison with its analogue from the established oxidation of Cj\u2010NCC\u20101.Pd\u2010NCC\u201054. Clearly, work\u2010up and preparation of extracts of cold senescent leaves need to be done swiftly, in order to avoid oxidation artefacts.Analysis of a minor NCC (tentatively named Prunus domestica ssp. domestica) were shown to contain a range of NCCs, two YCCs, and, in traces, a PiCC, all members of the \u2018type I\u2019 phyllobilin family. In spite of the absence of DCCs,Pd\u2010NCC\u201032 (1) showed a previously unknown structure and is functionalized with two glycopyranose moieties on the \u2018distant\u2019 pyrrole rings A and D. The structure of Pd\u2010NCC\u201032 (1) also provided the first (indirect) evidence for enzymatic glycosidation of an FCC at the 182\u2010position .Pd\u2010NCC\u201060 (5) and Pd\u2010NCC\u201071 (6) with corresponding Cj\u2010NCCs, indicated the plum NCCs to belong to the C(16)\u2010epi series, as well.Fig.\u00a0Extracts of naturally senescent leaves of the plum tree , LC/MS gradient grade MeOH from or VWR , and AcONH4, puriss. p.a., from Fluka . KH2PO4, puriss. p.a., K3PO4 dibasic\u2010anh., puriss. p.a., and hexane were from Sigma\u2013Aldrich . Sand was from J. T. Baker , Sep\u2010Pak\u00aeC18 cartridges (1 and 5\u00a0g) were from Waters Associates. pH Values were measured with a WTW SenTix 21 electrode connected to a WTW pH525 digital pH meter.HPLC grade MeOH was purchased from HPLC. Dionex Summit HPLC system with manual sampler, P680 pump, online degasser and diode array detector, 1.35\u00a0ml or 200\u00a0\u03bcl injection loop. Data were collected and processed with Chromeleon V6.70.i) Anal. HPLC. Kinetex 00G\u20104601\u2010E0\u20105u\u2010C18\u2010100A 250\u00a0\u00d7\u00a04.6\u00a0mm i.d. column at 20\u00a0\u00b0C protected with a Phenomenex AJ0\u20104287 C18 4\u00a0\u00d7\u00a03.0\u00a0mm i.d. pre\u2010column was used with a flow rate of 0.5\u00a0ml\u00a0min\u22121. Solvent A: 50mm aq. potassium phosphate buffer (pH 7.0), solvent B: MeOH, solvent C: H2O; solvent composition (A/B/C) as a function of time (0\u00a0\u2013\u00a090\u00a0min): 0\u00a0\u2013\u00a05, 80:20:0; 5\u00a0\u2013\u00a060, 80:20:0 to 40:60:0; 60\u00a0\u2013\u00a080, 40:60:0 to 0:100:0; 80\u00a0\u2013\u00a085, 0:100:0; 85\u00a0\u2013\u00a087, 0:100:0 to 0:20:80; 87\u00a0\u2013\u00a090, 0:20:80 to 80:20:0.ii) Semi\u2010prep. HPLC (90\u00a0min run). 00G\u20104252\u2010NO Luna 5u C18(2) 100A 250\u00a0\u00d7\u00a010\u00a0mm i.d. column at 20\u00a0\u00b0C protected with a Phenomenex AJ0\u20107220 250\u00a0\u00d7\u00a010\u00a0mm i.d. pre\u2010column was used with a flow rate as a function of time: 0\u00a0\u2013\u00a05\u00a0min: 1\u00a0\u2013\u00a04\u00a0ml\u00a0min\u22121; 5\u00a0\u2013\u00a090\u00a0min: 4\u00a0ml\u00a0min\u22121. Solvent A: 4mm aq. AcONH4, solvent B: MeOH with AcONH4 (c\u00a0=\u00a04mm), solvent C: H2O; solvent composition (A/B/C) as a function of time (0\u00a0\u2013\u00a090\u00a0min): 0\u00a0\u2013\u00a05, 80:20:0; 5\u00a0\u2013\u00a060, 80:20:0 to 40:60:0; 60\u00a0\u2013\u00a080, 40:60:0 to 0:100:0; 80\u00a0\u2013\u00a085, 0:100:0; 85\u00a0\u2013\u00a087, 0:100:0 to 0:20:80; 87\u00a0\u2013\u00a090, 0:20:80 to 80:20:0.iii) Semi\u2010prep. HPLC (70\u00a0min run). 00G\u20104252\u2010NO Luna 5u C18(2) 100A 250\u00a0\u00d7\u00a010\u00a0mm i.d. column at 20\u00a0\u00b0C protected with a Phenomenex AJ0\u20107220/1 C18 250\u00a0\u00d7\u00a010\u00a0mm i.d. pre\u2010column was used with a flow rate as a function of time: 0\u00a0\u2013\u00a05\u00a0min: 1\u00a0\u2013\u00a04\u00a0ml\u00a0min\u22121; 5\u00a0\u2013\u00a070\u00a0min: 4\u00a0ml\u00a0min\u22121. Solvent A: 50mm aq. potassium phosphate buffer (pH 7.0), solvent B: MeOH, solvent C: H2O; solvent composition (A/B/C) as function of time (0\u00a0\u2013\u00a070\u00a0min): 0\u00a0\u2013\u00a05, 80:20:0; 5\u00a0\u2013\u00a050, 80:20:0 to 47.3:52.7:0; 50\u00a0\u2013\u00a055, 47.3:52.7:0 to 0:52.7:47.3; 55\u00a0\u2013\u00a060, 0:52.7:47.3 to 0:100:0; 60\u00a0\u2013\u00a065, 0:100:0; 65\u00a0\u2013\u00a067, 0:100:0 to 0:20:80; 67\u00a0\u2013\u00a070, 0:20:80 to 80:20:0.LC/MS. i) Pre\u2010Purifcation of Minor Fractions Pd\u2010NCC\u201054 and Pd\u2010NCC\u201071 on an anal. HPLC. Minor catabolite fractions were first purified by HPLC according to following procedure: 3\u00a0\u2013\u00a04\u00a0g of leaf material were ground in mortar and pestle under liquid N2 with addition of ca. 1\u00a0g of sand, a tip of a spatula of CaCO3 and 4\u00a0\u2013\u00a05\u00a0ml of MeOH. The mixture was centrifuged and the supernatants were stored at \u221280\u00a0\u00b0C until use. An aliquot of the supernatant was centrifuged (1\u00a0min at 7200\u00a0g), diluted (1:1 v/v) with aq. potassium phosphate buffer and centrifuged again . In total 300\u00a0\u03bcl (3\u00a0\u00d7\u00a0100\u00a0\u03bcl) of the supernatant were purified on the anal. HPLC at a flow rate of 0.5\u00a0ml\u00a0min\u22121 using 50mm aq. K3PO4 as solvent A and MeOH as solvent B , and desired fractions were collected and combined.ii) LC/MS Analysis of minor fractions of the collected HPLC fractions were analyzed on an LC/MS system using AcONH4 buffer and MeOH as eluents . Twenty microliter of the collected catabolite fraction were injected and analyzed at a flow rate of 0.5\u00a0ml\u00a0min\u22121 .Spectroscopy. UV/VIS Spectra: Agilent Technologies Cary 60 spectrophotometer; \u03bbmax (nm) (rel. \u03b5). CD Spectra: Jasco J715, \u03bbmax and \u03bbmin (nm), \u0394\u03b5. 1H\u2010 and 13C\u2010NMR: Bruker 600\u00a0MHz Avance II+ (\u03b4(C1HD2OD) 3.31\u00a0ppm, and \u03b4(13CD3OD) 49.0\u00a0ppm, \u03b4 in ppm,J in Hz. Mass Spectrometry: Finnigan LCQ Classic, electrospray ionization (ESI) source, positive\u2010ion mode,Senescent plum tree leaves were harvested in November 2013 from a commercial orchard in Aldino (South Tyrol). They were immediately frozen in a freezer (\u221280\u00a0\u00b0C) and transported in a cold box (\u221220\u00a0\u00b0C) to Innsbruck, where they were stored cold (\u221280\u00a0\u00b0C).2) was frozen in liquid N2, grounded in a mortar and extracted with 1\u00a0ml of MeOH. The resulting suspension was centrifuged for 3\u00a0min at 13,000\u00a0g. Five hundred microliter of the MeOH supernatant were diluted with 2\u00a0ml of 50mm aq. potassium phosphate buffer (pH 7.0). After centrifugation for 3\u00a0min at 13,000\u00a0g, 200\u00a0\u03bcl of the extract was analyzed by HPLC . Yellow\u2010greenish senescent plum tree leaves (18.7\u00a0g) were frozen in liquid N2, pulverized to a fine powder and extracted with 60\u00a0ml of MeOH. The suspension was centrifuged for 5\u00a0min at 4000\u00a0g. Forty\u2010two milliliter of the supernatant were diluted with 168\u00a0ml of 50mm aq. potassium phosphate buffer (pH 7.0). After centrifugation for 5\u00a0min at 4000\u00a0g, the soln. was extracted two times with hexane. The MeOH extract was diluted with 300\u00a0ml of 50mm potassium phosphate buffer (pH 7.0) and applied to a pre\u2010conditioned 5\u00a0g SepPak cartridge. This was washed with 35\u00a0ml of H2O and the NCC\u2010containing fraction was eluted with 30\u00a0ml of MeOH. The solvents were removed by using a rotary evaporator. The residue was dissolved in 1\u00a0ml of MeOH and 4\u00a0ml of 50mm aq. potassium phosphate buffer (pH 7.0) using an ultrasonic bath. After centrifugation for 3\u00a0min at 13,000\u00a0g, the sample was divided in four aliquots and applied to semi\u2010prep. HPLC; injection volume, 1.25\u00a0ml; flow rate, 0\u00a0\u2013\u00a05\u00a0min: 1\u00a0\u2013\u00a04\u00a0ml\u00a0min\u22121, 5\u00a0\u2013\u00a090\u00a0min: 4\u00a0ml\u00a0min\u22121; solvent A: 4mm aq. AcONH4, solvent B: MeOH with AcONH4 (c\u00a0=\u00a04mm), solvent C: H2O; solvent composition (A/B/C) as a function of time (0\u00a0\u2013\u00a090\u00a0min): 0\u00a0\u2013\u00a05, 80:20:0; 5\u00a0\u2013\u00a060, 80:20:0 to 40:60:0; 60\u00a0\u2013\u00a080, 40:60:0 to 0:100:0; 80\u00a0\u2013\u00a085, 0:100:0; 85\u00a0\u2013\u00a087, 0:100:0 to 0:20:80; 87\u00a0\u2013\u00a090, 0:20:80 to 80:20:0. Fractions containing Pd\u2010NCC\u201032 (1) of five consecutive semi\u2010prep. HPLC runs were collected and dried under reduced pressure. The residue was dissolved in 200\u00a0\u03bcl of MeOH and 800\u00a0\u03bcl of 50mm aq. potassium phosphate buffer (pH 7.0) and re\u2010purified by semi\u2010prep. HPLC; injection volume, 1.00\u00a0ml; flow rate, 0\u00a0\u2013\u00a05\u00a0min: 1\u00a0\u2013\u00a04\u00a0ml\u00a0min\u22121, 5\u00a0\u2013\u00a070\u00a0min: 4\u00a0ml\u00a0min\u22121; solvent A: 50mm aq. potassium phosphate buffer (pH 7.0), solvent B: MeOH, solvent C: H2O; solvent composition (A/B/C) as a function of time (0\u00a0\u2013\u00a070\u00a0min): 0\u00a0\u2013\u00a05, 80:20:0; 5\u00a0\u2013\u00a050, 80:20:0 to 47.3:52.7:0; 50\u00a0\u2013\u00a055, 47.3:52.7:0 to 0:52.7:47.3; 55\u00a0\u2013\u00a060, 0:52.7:47.3 to 0:100:0; 60\u00a0\u2013\u00a065, 0:100:0; 65\u00a0\u2013\u00a067, 0:100:0 to 0:20:80; 67\u00a0\u2013\u00a070, 0:20:80 to 80:20:0. The fraction containing Pd\u2010NCC\u201032 (1) was collected between and diluted with 20\u00a0ml of 50mm aq. potassium phosphate buffer (pH 7.0). For de\u2010salting, the aq. soln. was applied to a pre\u2010conditioned 5\u00a0g SepPak cartridge, washed with 15\u00a0ml of H2O and eluted with 5\u00a0ml of MeOH. After removal of the solvents using a rotary evaporator, the sample was dried under high vacuum and a uniform sample of 0.29\u00a0mg of Pd\u2010NCC\u201032 (1) was obtained.Isolation of Raw Pd\u2010NCCs for Structural Analysis. 12 anal. extracts were prepared, combined and diluted with 95\u00a0ml of 50mm aq. potassium phosphate buffer (pH 7.0). This was applied to a pre\u2010conditioned 5\u00a0g SepPak cartridge, washed with 30\u00a0ml of H2O and the NCC\u2010containing fraction was eluted with 30\u00a0ml of MeOH. The fraction was dried under reduced pressure and the precipitate was dissolved in 400\u00a0\u03bcl of MeOH and 1.6\u00a0ml of 4mm aq. AcONH4. After centrifugation for 3\u00a0min at 13,000\u00a0g, the sample was divided in two aliquots and applied to semi\u2010prep. HPLC; injection volume, 1.00\u00a0ml; flow rate, 0\u00a0\u2013\u00a05\u00a0min: 1\u00a0\u2013\u00a04\u00a0ml\u00a0min\u22121, 5\u00a0\u2013\u00a090\u00a0min: 4\u00a0ml\u00a0min\u22121; solvent A: 4mm aq. AcONH4, solvent B: MeOH with AcONH4 (c\u00a0=\u00a04mm), solvent C: H2O; solvent composition (A/B/C) as a function of time (0\u00a0\u2013\u00a090\u00a0min): 0\u00a0\u2013\u00a05, 80:20:0; 5\u00a0\u2013\u00a060, 80:20:0 to 40:60:0; 60\u00a0\u2013\u00a080, 40:60:0 to 0:100:0; 80\u00a0\u2013\u00a085, 0:100:0; 85\u00a0\u2013\u00a087, 0:100:0 to 0:20:80; 87\u00a0\u2013\u00a090, 0:20:80 to 80:20:0. The fractions containing Pd\u2010NCC\u201032 (1), Pd\u2010NCC\u201035 (2), Pd\u2010NCC\u201040 (3), Pd\u2010NCC\u201056 (4) and Pd\u2010NCC\u201060 (5) were collected and to obtain pure samples from all fractions an anal. HPLC run with AcONH4 had to be performed; injection volume, 200\u00a0\u03bcl; flow rate, 0.5\u00a0ml\u00a0min\u22121; solvent A: 4mm aq. AcONH4, solvent B: MeOH with AcONH4 (c\u00a0=\u00a04mm), solvent C: H2O; solvent composition (A/B/C) as a function of time (0\u00a0\u2013\u00a090\u00a0min): 0\u00a0\u2013\u00a05, 80:20:0; 5\u00a0\u2013\u00a060, 80:20:0 to 40:60:0; 60\u00a0\u2013\u00a080, 40:60:0 to 0:100:0; 80\u00a0\u2013\u00a085, 0:100:0; 85\u00a0\u2013\u00a087, 0:100:0 to 0:20:80; 87\u00a0\u2013\u00a090, 0:20:80 to 80:20:0. In each anal. HPLC run, the desired catabolite was collected.Pd\u2010NCC\u201032 (1). tR\u00a0=\u00a032.6\u00a0min. UV/VIS : 244sh (0.83), 314 (1.00) : 226 (8), 249 (\u22123), 263 (\u22123), 283 (\u22128), 319 (1). 1H\u2010NMR : 1.92 ); 2.08 ); 2.12 ); 2.23 ); 2.26\u00a0\u2013\u00a02.30 ); 2.31\u00a0\u2013\u00a02.35 ); 2.37\u00a0\u2013\u00a02.41 ); 2.60\u00a0\u2013\u00a02.65 , Ha\u2013C(121)); 2.71\u00a0\u2013\u00a02.77 ); 2.91 ); 3.17 ); 3.21 ); 3.24\u00a0\u2013\u00a03.28 , H\u2013C(4\u2033), H\u2013C(5\u2032), H\u2013C(5\u2033)); 3.33\u00a0\u2013\u00a03.39 , H\u2013C(3\u2032), H\u2013C(3\u2033)); 3.62\u00a0\u2013\u00a03.69 , Ha\u2013C(6\u2033), Hb\u2013C(32)); 3.71 ); 3.75 ); 3.83\u00a0\u2013\u00a03.87 , Hb\u2013C(6\u2033)); 3.96\u00a0\u2013\u00a04.01 , Hb\u2013C(182)); 4.02\u00a0\u2013\u00a04.09 , Hb\u2013C(5)); 4.17 ); 4.33 ); 4.77 ); 4.87 ); 9.32 ). 13C\u2010NMR : 8.4 (C(21)); 9.1 (C(71)); 9.1 (C(131)); 12.5 (C(171)); 22.0 (C(121)); 23.6 (C(5)); 24.8 (C(31)); 29.7 (C(15)); 36.9 (C(10)); 39.6 (C(122)); 52.7 (C(85)); 62.1 (C(6\u2032)); 62.1 (C(6\u2033)); 62.5 (C(16)); 66.6 (C(181)); 67.7 (C(82)); 70.3 (C(32)); 71.1 (C(4\u2032)); 71.1 (C(4\u2033)); 73.2 (C(182)); 74.8 (C(2\u2032)); 74.8 (C(2\u2033)); 77.7 (C(3\u2032)); 77.7 (C(3\u2033)); 77.7 (C(5\u2032)); 77.7 (C(5\u2033)); 103.9 (C(1\u2032)); 103.9 (C(1\u2033)); 112.2 (C(7)); 114.8 (C(13)); 120.0 (C(3)); 120.0 (C(12)); 124.2 (C(11)); 124.2 (C(14)); 125.6 (C(8)); 128.5 (C(1)); 133.9 (C(6)); 134.0 (C(2)); 139.6 (C(4)); 159.3 (C(17)); 130.2 (C(18)); 161.5 (C(9)); 171.4 (C(83)); 174.8 (C(19)); 177.2 (C(20)). ESI\u2010MS: 1079.2 ; 1063.3 ; 1047.3 ; 1041.3 ; 1025.3 ; 1005.1 (20), 1004.1 (53), 1003.1 ; 987.3 (7); 879.3 ; 863.4 ; 841.2 ; 827.3 (4); 684.1 ; 679.2 .see Fig.\u00a0. CD (MeOPd\u2010NCC\u201035 (2). tR\u00a0=\u00a035.5\u00a0min. UV/VIS (4mm aq. AcONH4/MeOH 63:37): 284 (0.75), 316 (1.00). ESI\u2010MS: 879.3 ; 863.3 ; 843.2 (15), 842.2 (47), 841.2 ; 825.3 (6); 809.3 ; 684.2 ; 679.2 ; 522.1 .Pd\u2010NCC\u201040 (3). tR\u00a0=\u00a040.6\u00a0min. UV/VIS (4mm aq. AcONH4/MeOH 59:41): 278 (0.78), 316 (1.00). ESI\u2010MS: 755.1 ; 739.2 ; 717.3 ; 701.3 ; 681.2 (10), 680.1 (40), 679.2 ; 647.2 ; 522.1 .Pd\u2010NCC\u201054. UV/VIS (4mm aq. AcONH4/MeOH 40:60): 316\u00a0nm. ESI\u2010MS: 699.2 ; 678.2 ; 663 (4), 662 (32), 661.2 ; 629.3 .Pd\u2010NCC\u201056 (4). tR\u00a0=\u00a056.4\u00a0min. UV/VIS (4mm aq. AcONH4/MeOH 47:53): 316\u00a0nm. ESI\u2010MS: 845.3 ; 829.3 ; 809.2 (15), 808.2 (46), 807.2 ; 775.3 ; 684.2 ; 645.2 .Pd\u2010NCC\u201060 (5). tR\u00a0=\u00a060.0\u00a0min. UV/VIS (4mm aq. AcONH4/MeOH 44:56): 315 (1.00). ESI\u2010MS: 683.2 ; 667.3 ; 647.2 (11), 646.2 (39), 645.2 ; 613.2 ; 522.1 .Pd\u2010NCC\u201060 (5) and Cj\u2010NCC\u20101Pd\u2010NCC\u201060 (5), of Cj\u2010NCC\u20101, as well as a 1:1 mixture of both were analyzed by anal. HPLC . tR\u00a0=\u00a070.7\u00a0min. UV/VIS (50mm aq. potassium phosphate buffer (pH 7.0)/MeOH 20:80): 239sh (1.00), 316 (0.87). ESI\u2010MS: 667.1 ; 631 (2), 630 (35), 629.2 ; 597.2 ; 506 . Provisional identification of Pd\u2010NCC\u201071 (6) with Cj\u2010NCC\u20102Pd\u2010NCC\u201071 (6), a separate sample of purified Cj\u2010NCC\u20102, as well as a mixture of both were analyzed by anal. HPLC.Pd\u2010YCC\u201061. tR\u00a0=\u00a061.3\u00a0min. UV/VIS (50mm aq. potassium phosphate buffer (pH 7.0)/MeOH 40:60): 246 (0.73), 313 (1.00), 429 (1.77). ESI\u2010MS: 881.1 ; 865.3 ; 843.1 ; 827.3 ; 807.2 (17), 806.1 (48), 805.1 ; 796.6 (16); 774.4 (26); 756.3 (17); 700.3 (15); 643.1 ; 611.3 .Pd\u2010YCC\u201067. tR\u00a0=\u00a067.1\u00a0min. UV/VIS (50mm aq. potassium phosphate buffer (pH 7.0)/MeOH) 25:75): 247 (0.74), 316 (1.00), 428 (1.29). ESI\u2010MS: 719.1 ; 703.2 ; 681.1 ; 665.3 ; 645.2 (11), 644.2 (40), 643.2 ; 611.2 .Pd\u2010PiCC\u201075. tR\u00a0=\u00a075.0\u00a0min. UV/VIS (50mm aq. potassium phosphate buffer (pH 7.0)/MeOH 1:9): 314 (0.75), 525 (1.00).Pd\u2010PiCC\u201075 (in an extract of a plum tree leaf) and of a purified sample of Cj\u2010PiCCProvisional identification of"} +{"text": "Glycosylation of the immunoglobulin G (IgG)-Fc tail is required for binding to Fc-gamma receptors (Fc\u03b3Rs) and complement-component C1q. A variety of IgG1-glycoforms is detected in human sera. Several groups have found global or antigen-specific skewing of IgG glycosylation, for example in autoimmune diseases, viral infections, and alloimmune reactions. The IgG glycoprofiles seem to correlate with disease outcome. Additionally, IgG-glycan composition contributes significantly to Ig-based therapies, as for example IVIg in autoimmune diseases and therapeutic antibodies for cancer treatment. The effect of the different glycan modifications, especially of fucosylation, has been studied before. However, the contribution of the 20 individual IgG glycoforms, in which the combined effect of all 4 modifications, to the IgG function has never been investigated. Here, we combined six glyco-engineering methods to generate all 20 major human IgG1-glycoforms and screened their functional capacity for Fc\u03b3R and complement activity. Bisection had no effect on Fc\u03b3R or C1q-binding, and sialylation had no- or little effect on Fc\u03b3R binding. We confirmed that hypo-fucosylation of IgG1 increased binding to Fc\u03b3RIIIa and Fc\u03b3RIIIb by ~17-fold, but in addition we showed that this effect could be further increased to ~40-fold for Fc\u03b3RIIIa upon simultaneous hypo-fucosylation and hyper-galactosylation, resulting in enhanced NK cell-mediated antibody-dependent cellular cytotoxicity. Moreover, elevated galactosylation and sialylation significantly increased (independent of fucosylation) C1q-binding, downstream complement deposition, and cytotoxicity. In conclusion, fucosylation and galactosylation are primary mediators of functional changes in IgG for Fc\u03b3R- and complement-mediated effector functions, respectively, with galactose having an auxiliary role for Fc\u03b3RIII-mediated functions. This knowledge could be used not only for glycan profiling of clinically important (antigen-specific) IgG but also to optimize therapeutic antibody applications. The importance of the biological properties of antibodies to specifically engage a target of choice and activate complement and Fc gamma receptors (Fc\u03b3R) on immune cells is curreThis fucose residue is part of a conserved glycan on asparagine 297 in the Fc domain of immunoglobulin G (IgG). This glycan is important for the quaternary structure of the Fc part, since its removal abrogates binding of Fc\u03b3R and C1q and hence the antibody\u2019s effector functions , 11. ThiN-acetylglycosamines and mannoses and can be found in human serum with variable levels of core fucose, bisecting N-acetylglycosamine, galactose, and terminal sialic acids or RBCs. NK cell isolation was only performed with blood from well-genotyped donors who do not express Fc\u03b3RIIc to excluEscherichia coli strain DH5\u03b1 was used for recombinant DNA work. Restriction endonucleases, DNA modification enzymes were obtained from Thermo Fisher Scientific . Oligonucleotides were obtained from Geneart (Thermo Fisher Scientific) or Integrated DNA Technologies .Variable (V) genes for anti-human RhD (anti-D clone 19A10) heavy and light chain were sequenced from a single human B cell from a hyper immunized donor . A singll-fucose (2FF) or 1\u2009mM 2-deoxy-2-fluoro-d-galactose (2FG) (Carbosynth), respectively, was added to the cell suspension 4\u2009h post transfection. To increase bisecting GlcNAc, 1% pEE6.4\u2009+\u2009GNTIII encoding mannosyl -glycoprotein beta-1,4-N-acetylglucosaminyltransferase (GNTIII) enzyme was co-transfected with 99% IgG1-\u03ba HC\u2009+\u2009LC vector. To increase galactose, 1% pEE6.4\u2009+\u2009B4GALT1 encoding \u03b2-1,4-galactosyltransferase 1 (B4GALT1) enzyme was co-transfected with 99% IgG1 vector and 5\u2009mM d-galactose was added to the cell suspension 1\u2009h before transfection. To increase sialylation, the level of galactosylation must also be elevated as sialic acid is the terminal sugar group with galactose residues as substrate. Thus, 1% pEE6.4\u2009+\u2009B4GALT1 and 2.5% pEE14.4\u2009+\u2009STGALT encoding \u03b2-galactoside alpha-2,6-sialyltransferase 1 (ST6GALT) were both co-transfected 96.5% IgG1 vector and 5\u2009mM d-galactose was added to the cell suspension 1\u2009h before transfection. To further increase sialylation, in vitro sialylation (ivs) was performed on the purified in vivo sialylated IgG created using the previous method. Recombinant human \u03b1-2,6-sialyltransferase and cytidine-5\u2032-monophospho-N-acetylneuraminic acid (CMP-NANA) (Roche) were incubated at 37\u00b0C for 24\u2009h with purified IgG1 with already in vivo enhanced galactose and sialic acid (as described above), after incubation samples were re-purified with protein A, as described previously (IgG1 production in human embryonic kidney (HEK) F cells and purification using protein A affinity chromatography was performed as described previously by Kruijssen et al. Glyco-eneviously , 34.via a CaptiveSpray source with a NanoBooster . The latter enriched the N2 flow (3\u2009l/min) with CH3CN (pressure 0.2\u2009bar), resulting in increased sensitivity. The samples were ionized in positive ion mode at 1,100\u2009V. The Maxis Impact quadrupole-TOF\u2013MS was used as detector. MS1 spectra were collected at a frequency of 1\u2009Hz with a scan range of m/z 550\u20131,800. The mass spectrometric data were calibrated internally in DataAnalysis 4.0 using a list of known IgG glycopeptide masses. MSConvert (Proteowizard 3.0) (m/z window of \u00b10.2 and within a time window of \u00b115\u2009s surrounding the retention time) was extracted using the in-house developed 3D Max Xtractor software tool. If above a signal:background ratio of 3, the background-subtracted area of the first three isotopic peaks of each glycopeptide in both 2+, 3+, and 4+ charge state were summed, and this summed value was then divided by the total summed value of all IgG1 glycopeptides to arrive at a percentage for each glycopeptide. From these percentages, we calculated several derived traits using the following formulas: fucosylation (H3N3F1\u2009+\u2009H4N3F1\u2009+\u2009H5N3F1\u2009+\u2009H6N3F1\u2009+\u2009G0F\u2009+\u2009G1F\u2009+\u2009G2F\u2009+\u2009H6N4F1\u2009+\u2009G0FN\u2009+\u2009G1FN\u2009+\u2009G2FN\u2009+\u2009H6N5F1\u2009+\u2009H4N3F1S1\u2009+\u2009H5N3F1S1\u2009+\u2009H6N3F1S1\u2009+\u2009G1FS\u2009+\u2009G2FS\u2009+\u2009H6N4F1S1\u2009+\u2009G2FS2\u2009+\u2009G1FNS\u2009+\u2009G2FNS\u2009+\u2009H6N5F1S1\u2009+\u2009G2FNS2), bisection (H6N4F1\u2009+\u2009G0FN\u2009+\u2009G1FN\u2009+\u2009G2FN\u2009+\u2009H6N5F1\u2009+\u2009H6N4F1S1\u2009+\u2009G1FNS\u2009+\u2009G2FNS\u2009+\u2009H6N5F1S1\u2009+\u2009G2FNS2\u2009+\u2009H6N4\u2009+\u2009G0N\u2009+\u2009G1N\u2009+\u2009G2N\u2009+\u2009H6N5\u2009+\u2009H6N4S1\u2009+\u2009G1NS\u2009+\u2009G2NS\u2009+\u2009H6N5S1\u2009+\u2009G2NS2), galactosylation [(H4N3F1\u2009+\u2009H5N3F1\u2009+\u2009G1F\u2009+\u2009H6N4F1\u2009+\u2009G1FN\u2009+\u2009H6N5F1\u2009+\u2009H4N3F1S1\u2009+\u2009H5N3F1S1\u2009+\u2009H6N3F1S1\u2009+\u2009G1FS\u2009+\u2009H6N4F1S1\u2009+\u2009G1FNS\u2009+\u2009H6N5F1S1\u2009+\u2009H4N3\u2009+\u2009H5N3\u2009+\u2009H6N3\u2009+\u2009G1\u2009+\u2009H6N4\u2009+\u2009G1N\u2009+\u2009H6N5\u2009+\u2009H4N3S1\u2009+\u2009H5N3S1\u2009+\u2009H6N3S1\u2009+\u2009G1S\u2009+\u2009H6N4S1\u2009+\u2009G1NS\u2009+\u2009H6N5S1) * 0.5\u2009+\u2009G2F\u2009+\u2009G2FN\u2009+\u2009G2FS\u2009+\u2009G2FS2\u2009+\u2009G2FNS\u2009+\u2009G2FNS2\u2009+\u2009G2\u2009+\u2009G2N\u2009+\u2009G2S\u2009+\u2009G2S2\u2009+\u2009G2NS\u2009+\u2009G2NS2], sialylation [(H4N3F1S1\u2009+\u2009H5N3F1S1\u2009+\u2009H6N3F1S1\u2009+\u2009G1FS\u2009+\u2009G2FS\u2009+\u2009H6N4F1S1\u2009+\u2009G1FNS\u2009+\u2009G2FNS\u2009+\u2009H6N5F1S1\u2009+\u2009H4N3S1\u2009+\u2009H5N3S1\u2009+\u2009H6N3S1\u2009+\u2009G1S\u2009+\u2009G2S\u2009+\u2009H6N4S1\u2009+\u2009G1NS\u2009+\u2009G2NS\u2009+\u2009H6N5S1) * 0.5\u2009+\u2009G2FS2\u2009+\u2009G2FNS2\u2009+\u2009G2S2\u2009+\u2009G2NS2], hybrid-types (H5N3F1\u2009+\u2009H6N3F1\u2009+\u2009H6N4F1\u2009+\u2009H6N5F1\u2009+\u2009H5N3F1S1\u2009+\u2009H6N3F1S1\u2009+\u2009H6N4F1S1\u2009+\u2009H6N5F1S1\u2009+\u2009H5N3\u2009+\u2009H6N3\u2009+\u2009H6N4\u2009+\u2009H6N5\u2009+\u2009H5N3S1\u2009+\u2009H6N3S1\u2009+\u2009H6N4S1\u2009+\u2009H6N5S1), and high-mannose (H5N2\u2009+\u2009H6N2\u2009+\u2009H7N2\u2009+\u2009H8N2\u2009+\u2009H9N2). For some of the minor hybrid-type glycans, it could not be determined conclusively whether a galactose or a bisecting N-acetylglucosamine was present, so an educated guess was made based on structural knowledge .Immunoglobulin G-Fc glycan composition of produced IgG1 was determined by mass spectrometry as described previously by Dekkers et al. Trypsin-ard 3.0) was usedard 3.0) was usedProtein A purified IgG was analyzed for monomeric and dimeric IgG on a Superdex 200 10/300 gel filtration column connected to an \u00c4kta explorer HPLC system at RT with a flow rate of 0.5\u2009ml/min and PBS as running buffer. Elution profiles were obtained by recording the absorbance at 215\u2009nm.Human Fc\u03b3R constructs for surface plasmon resonance (SPR) analysis were obtained from Sino biological . To further include all human Fc\u03b3Rs, a fusion Fc\u2013Fc\u03b3R construct composed of the extracellular domain of the Fc\u03b3RIIIb in both allotypes followed by a Fc domain was created. To create the fusion Fc\u2013Fc\u03b3RIIIb constructs the amino acid code of the extracellular domain of either Fc\u03b3RIIIb of NA1 allotype or Fc\u03b3RIIIb NA2 allotype NCBI re. For bioKD) was done using an equilibrium analysis by linear intrapolation to Rmax\u2009=\u2009500 at 109\u2009cells/ml. An amount of 105 erythrocytes were incubated with NK cells for 2\u2009h at 37\u00b0C in a 2:1 ratio in Iscove\u2019s modified dulbecco\u2019s medium supplemented with 10% fetal calf serum and anti-D IgG1-glycoforms at a total volume of 100\u2009\u00b5l. To determine 100% lysis, 2.5% saponine was added to RBC in control wells and spontaneous lysis (sp) was determined by incubation of RBC without NK cells. Supernatants were collected and released 51Cr was quantified in a Packard Cobra II Auto-Gamma Counter Model D5005 (PerkinElmer). Percentage cytotoxicity was determined by the following formula: NK cells were isolated from Ficoll-Plaque\u2122-Plus gradient obtained PBMCs by a CD56 magnetic-activated cell separation isolation kit , according to manufacturer\u2019s description. D\u2009+\u2009RBCs were isolated and labeled with radioactive chromium and incubated 30\u2009min at RT. To remove unbound TNBS, the solution was dialyzed using a dialysis cassette for 1.5\u2009h at RT against PBS and additionally overnight at 4\u00b0C to obtain HSA-TNP.A 2.4-mM 2,4,6-trinitrobenzenesulfonic acid (TNBS) (Sigma-Aldrich) solution was added to 20\u2009mg human serum albumin (HSA) diluted to 20\u2009mg/ml in 0.2\u2009M NA+/+/plx {veronalbuffer \u2009+\u200910\u2009mM CaCl2 (Merck)\u2009+\u20092\u2009mM MgCl2 (Merck)\u2009+\u20090.1% poloxamer} for 1\u2009h at RT. When C1q was blocked, 10\u2009min prior to addition of serum to the ELISA plate, anti-C1q-85 blocking antibody and the reaction was stopped with the addition of 100\u2009\u00b5l 2\u2009M H2SO4 (Merck). The optical density (OD) was measured at A450\u2009nm using a plate reader .To coat, maxisorp plates were incubated o/n at RT with 20\u2009\u00b5g/ml HSA-TNP in PBS. The plates were washed 5\u00d7 with PBS\u2009+\u20090.1% tween-20 (Sigma-Aldrich) (wash buffer) using an ELISA washer . All following washing steps were done similarly. The IgG samples were diluted in 100\u2009\u00b5l PBS/plx [PBS\u2009+\u20090.1% poloxamer ] per well and incubated for 1.5\u2009h at RT. The plates were washed and incubated with 100\u2009\u00b5l 1:35 serum pool in VBantibody was addeti-C1q-2 , 0.5\u2009\u00b5g/ti-C1q-2 , 0.6\u2009\u00b5g/ti-C1q-2 , or 1\u2009\u00b5gThe results were analyzed with a parallel line assay in Microsoft Office Excel . We asse+ RBCs obtained from heparinized blood were mixed with 350\u2009\u00b5l 0.313\u2009mM TNBS in 0.15\u2009M Na2HPO4, pH 8.8 and incubated for 10\u2009min at RT. TNPylated RBCs were centrifuged for 2\u2009min at 350\u2009\u00d7\u2009g and washed two times with PBS. RBC were resuspended into VBG+/+ [VB+/+\u2009+\u20090.05% w/v gelatin (Sigma-Aldrich)]. Anti-TNP IgG1 was serially diluted in VBG\u2212/\u2212 . In round bottom plates to a final volume of 100\u2009\u00b5l we combined the diluted IgG1, 10% serum, ~4.5\u2009\u00d7\u2009106 RBC, and a glass bead to ensure mixing of the solution during incubation . This amount of RBC was taken to ensure the 100% absorbance between 1.8 and 2.2 delta (\u0394) A412\u2013A690\u2009nm. The plates were incubated for 90\u2009min at 37\u00b0C while shaking at 150\u2009rpm . After incubation, 1.25% w/v saponine was supplemented to the 100% control wells, 100\u2009\u00b5l VBG\u2212/\u2212 was added to all wells and the plates were centrifuged for 2\u2009min at 350\u2009\u00d7\u2009g. Subsequently, 150\u2009\u00b5l of supernatant was transferred into a separate plate and OD was measured at \u0394 A412\u2013A690\u2009nm using a plate reader. The percentage of lysed cells was calculated as follows: 50) for each replicate of the different glycoforms using a non-linear fit for normalized response with a variable slope and combined these to an average EC50.Fifty microliters of washed, packed, Dp\u2009<\u20090.05 using two-tailed tests.Statistical analyses were performed using GraphPad Prism version 6.00 for Windows . The level of significance was set at x-axis legend, Figures N-acetylglucosaminyltransferase III (GntIII) to increase bisection \u2014but this was only observed in samples with low starting-levels of galactosylation Figure , 47, 48.) Figure , with boKD of more than twofold from unmodified IgG to be potentially meaningful changes and within the scope of the SPR method, using a simplified 1:1 Langmuir model that does not fully represent the actual interaction which is more complicated and subsequent C4b deposition was titrated by serial dilution . All glycovariants of anti-TNP bound TNP-HSA equally well , but C1q binding and C4b deposition differed profoundly for the different glycoforms . The relative C1q binding and C4b deposition were then calculated Figures A and witWe have previously created an orthogonal set of glyco-engineering tools which weOf the Fc\u03b3Rs, we only observed an effect of glycosylation on binding to the Fc\u03b3RIII-family of receptors, both Fc\u03b3RIIIa and Fc\u03b3RIIIb and their allotypes, which confirms and expands recent studies using a limited set of glycovariants presented here , 30. IncThe enhanced binding of galactosylated and afucosylated IgG was slightly weakened by addition of sialic acid, but only if a bisecting GlcNAc was present. A similar negative effect of sialylation has previously been observed for mouse Fc\u03b3R by Ravetch and colleagues . ImportaIn vitro, this has been found result in stronger functional efficacy for the V158-variant . In vitr-variant \u201357. In vlearance . It shoulearance . We alsolearance , 62, 63.Importantly, the observed changes in Fc\u03b3RIIIa-binding due to glycosylation reliably translated into functional NK cell-mediated ADCC lysis of RBC. For Fc\u03b3RIIIa and Fc\u03b3RIIIb it was known that absence of IgG-Fc core-fucosylation increases the affinity of interaction due to a glycan\u2013glycan interaction between the Fc glycan and the N162-glycan uniquely found in the Fc\u03b3RIII family . Our appThe possible effect of the Fc-glycans on complement activity, has until now remained enigmatic. It has been proposed for a long time that agalactosylated IgG activates complement more efficiently through the lectin pathway (MBL) . To our Low galactosylation level in total IgG generally correlates with disease severity of several autoimmune diseases, such as rheumatoid arthritis and multiple sclerosis , 14. WhiIn summary, we show here that a set of glyco-engineering techniques we recently developed can be cPeripheral blood from anonymous, healthy volunteers was obtained with informed, written consent of all subjects, in accordance with Dutch regulations. This study was approved by the Sanquin Ethical Advisory Board in accordance with the Declaration of Helsinki.GD, TK, DW, TR, MW, and GV designed the research. GD, AB, DW, TR, MW, and GV designed the experiments. GD, LT, RP, AB, MdB, CK, SL-T, RV, and YM performed the experiments. GD, RP, AB, MdB, CK, TR, TK, MW, and GV analyzed data, GD and GV wrote the manuscript. All authors contributed to and approved the final manuscript.The authors declare that this study received funding from Sanquin Bloedvoorziening, a not-for-profit organization. The funder was not involved in the study design or collection, analysis, or interpretation of the data."} +{"text": "The fused oxazine ring adopts an approximately half-chair conformation. The two benzoxazine rings are oriented anti to one another around the central C\u2014C bond. The dominant inter\u00admolecular inter\u00adaction in the crystal structure is a C\u2014H\u22efF hydrogen bond between the F atoms and the axial H atoms of the OCH2N methyl\u00adene group in the oxazine rings of neighbouring mol\u00adecules. C\u2014H\u22ef\u03c0 contacts further stabilize the crystal packing.The title fluorinated bis\u00adbenzoxazine, C Bond lengths in the benzoxazine moiety in (I)et al., 2012abThe mol\u00adecular structure of the title compound is illustrated in Fig.\u00a01Q = 0.4913\u2005(15)\u2005\u00c5, and \u03b8 = 52.03\u2005(17)\u00b0 and \u03c6 = 98.3\u2005(2)\u00b0, with C2 and N1 displaced from the mean plane by \u22120.299\u2005(2) and 0.331\u2005(1)\u2005\u00c5, respectively. The C1\u2014C1A bond is in an axial position with a C5\u2014N1\u2014C1\u2014C1A torsion angle of 75.45\u2005(18)\u00b0. The two benzoxazine rings are oriented anti to one another about the central C1\u2014C1A bond, with an N1\u2014C1\u2014C1A\u2014N1A torsion angle of 180.0\u2005(2)\u00b0.The fused six-membered heterocyclic rings exist in an approximately half-chair conformation, characterized by a puckering amplitude A\u22efF1 hydrogen bonds and 3.577\u2005(2)\u2005\u00c5 and with C\u2014H\u22efCg angles of 126 and 129\u00b0, respectively.The packing of title compound is dominated by C2\u2014H2s Table\u00a01, that cos Table\u00a01, as indiH-1,3-benzoxazine) monohydrate bis set to 1.2Ueq of the parent atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016015243/sj5506sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016015243/sj5506Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016015243/sj5506Isup3.cmlSupporting information file. DOI: 1507056CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The structure of the title squarate salt is reported. Classical N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds combine with weak C\u2014O\u22ef\u03c0(ring) and \u03c0\u2013\u03c0 contacts to stabilize the crystal packing. 6H10N22+\u00b72C4HO4\u2212, comprises two hydrogen squarate anions and a 2-(aza\u00adniumylmeth\u00adyl)pyridinium dication. The squaric acid mol\u00adecules each donate an H atom to the N atoms of the pyridine ring and the amino\u00admethyl units of a 2-(amino\u00admeth\u00adyl)pyridine mol\u00adecule, forming the 1:2 salt. The Hsq\u2212 anions are linked by strong O\u2014H\u22efO hydrogen bonds and an N\u2014H\u22efO hydrogen bond links the 2-(aza\u00adniumylmeth\u00adyl)pyridinium cation to one of the squaric acid anions. The crystal structure features additional N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, \u03c0\u2013\u03c0 stacking and unusual weak C\u2014O\u22ef\u03c0(ring) inter\u00adactions.The asymmetric unit of the title compound, C It can be found in three forms: uncharged H2sq, the Hsq\u2212 monoanion and the sq2\u2212 dianion. The mono- and dianions are often produced following deprotonation by amines electron donors of one or two lone pairs of electrons. Recently, we reported the synthesis and characterization of the same organo\u00adammonium squarate as the title compound but as a hydrate in the triclinic space group PP21/c.Hydrogen bonding is the most common way of generating supra\u00admolecular organic systems in crystal engineering and mol\u00adecular recognition. Hydrogen-bonded systems generated from organic cations and anions are of special inter\u00adest as they would be expected to form stronger hydrogen bonds than those in neutral mol\u00adecules \u2005\u00c5] hydrogen bond pyridine forms a salt with two squaric acid mol\u00adecules and each mol\u00adecule of the acid loses one proton. One of these is transferred to the N atom of the pyridine ring, generating the 4-(amino\u00admeth\u00adyl)morpholinium mono-cation. The other from the second acid mol\u00adecule is engaged in the formation of a homo-conjugated hydrogen squarate anion nd Fig.\u00a01. The elend Fig.\u00a01. ConsideCg1, Cg2 and Cg3. The dihedral angles between the mean plane of ring 1 and those of rings 2 and 3 are 18.818\u2005(8) and 31.564\u2005(6)\u00b0, respectively. The dihedral angle between the two squarate anions is 29.19\u2005(1)\u00b0. The angles between the C\u2014C bonds in the Hsq\u2212 anions are close to 90\u00b0, with the oxygen atoms directed almost along the diagonals.N1/C1\u2013C5, C7\u2013C10 and C11\u2013C14 are defined as rings 1, 2 and 3, respectively, with centroids \u2212 ion has one C\u2014O bond (C11\u2014O5) at 1.3023\u2005(17)\u2005\u00c5, which is significantly longer than a normal single C\u2014O bond. This most likely reflects the involvement in the negative charge-assisted hydrogen bonding mentioned earlier. At 1.3000\u2005(15)\u2005\u00c5, the C10\u2014O4 bond is similarly extended. The remaining C\u2014O bonds in both rings display a similar pattern with one obvious C=O double bond in each ring and the others of inter\u00admediate length in the range 1.2356\u2005(16) to 1.2658\u2005(15)\u2005\u00c5, indicating some delocalization occurring in both rings.The C\u2014C distances in the planar squarate ring systems reflect partial double-bond character for C9\u2014C10, C7\u2014C10, C11\u2014C12 and C11\u2014C14 with distances of 1.4291\u2005(17), 1.4357\u2005(16), 1.4139\u2005(17) and 1.4465\u2005(18)\u2005\u00c5, respectively. In contrast C7\u2014C8, C8\u2014C9, C12\u2014C13 and C13\u2014C14 display more single-bond character with distances of 1.4886\u2005(17), 1.4929\u2005(17), 1.4802\u2005(18) and 1.5141\u2005(17)\u2005\u00c5, respectively. The HsqA\u22efO3 [2.4583\u2005(14)\u2005\u00c5] related to the proton-sharing inter\u00adaction discussed earlier. This pair of anions is further linked to the 2-(aza\u00adniumylmeth\u00adyl)pyridinium dication by an N1\u2014H1A\u22efO1 hydrogen bond, Fig.\u00a01A\u22efO3, N2\u2014H2B\u22efO2i and N2\u2014H2B\u22efO5i hydrogen bonds form rings with an C\u22efO6i and N2\u2014H2B\u22efO5i hydrogen bonds combine to form C\u22efO6i, and N2\u2014H2A\u22efO8ii and homonuclear O4\u2014H4A\u22efO6iii and O5\u2014H5A\u22efO3 hydrogen bonds generate a larger Cg2ii, C7\u2014O1\u22efCg3iv and C13\u2014O7\u22efCg2v inter\u00adactions reinforced by \u03c0\u2013\u03c0 stacking inter\u00adactions. These latter contacts also contribute to the stabilization of the crystal packing and 2-(amino\u00admeth\u00adyl)pyridine were dissolved in water (25\u2005cm3) to obtain a mixture in the molar ratio 2:1 and the solution was heated to 323\u2005K in a temperature-controlled bath and stirred for one\u2005h. The reaction mixture was then slowly cooled to room temperature. The crystals formed were filtered and washed with 10\u2005cm3 of methanol, and dried in air.All chemical reagents were analytical grade commercial products. The solvent was purified by conventional methods. Squaric acid (HUiso(H) = 1.2Ueq(C). The NH3 group (N\u2014H = 0.89\u2005\u00c5) was also allowed to ride in the refinement with Uiso(H) = 1.5Ueq(N). The O-bound H atoms and N1-bound H atom were located in a difference-Fourier map and refined with Uiso(H) = 1.2Ueq(O) and Uiso(H) = 1.5Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017004376/sj5524sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017004376/sj5524Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017004376/sj5524Isup3.cmlSupporting information file. DOI: 1538918CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The macrocyclic title compound crystallizes as a dihydrate with a 12-membered inner ring system. Hydrogen bonds involving the lattice water mol\u00adecules link the components into a three-dimensional network system. 21H40N4\u00b72H2O, has been determined from synchrotron X-ray radiation data. The asymmetric unit comprises one 12-membered macropolycycle and two lattice water mol\u00adecules. The macropolycycle contains two cyclo\u00adhexane rings and one 1,3-di\u00adaza\u00adcyclo\u00adhexane ring, all in chair conformations. The C\u2014N and C\u2014C bond lengths are in the ranges 1.4526\u2005(16)\u20131.4786\u2005(17) and 1.517\u2005(2)\u20131.5414\u2005(17)\u2005\u00c5, respectively. One intra\u00admolecular N\u2014H\u22efN hydrogen bond helps to stabilize the mol\u00adecular conformation while medium-strength inter\u00admolecular N\u2014H\u22efO, O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds involving the lattice water mol\u00adecules connect the components into a three-dimensional network.The structure of the title compound, C The two methyl substituents of the C10 and C20 atoms are on the same side with respect to the macrocyclic plane of the four N atoms \u20131.4786\u2005(17)\u2005\u00c5 and 1.517\u2005(2)\u20131.5414\u2005(17)\u2005\u00c5 for the C\u2014N and C\u2014C bonds, respectively. The N1\u2014C20 distance is the longest C\u2014N distance, presumably as a consequence of the methyl group on the C20 atom and the N\u22efH\u2014O hydrogen bond involving N1. The bond angles within the six-membered 1,3-di\u00adaza\u00adcyclo\u00adhexane ring, N2\u2014C7\u2014N3, C7\u2014N2\u2014C8, and C7\u2014N3\u2014C10, are 109.89\u2005(10), 109.60\u2005(10), and 108.08\u2005(9)\u00b0, respectively. All other C\u2014N, C\u2014C, and C\u2014H bond lengths and corresponding angles are in the normal range for such compounds unit, viz. the crystal structure of [Cr(L2)(H2O)](ClO4)2\u00b73H2O docosane (L1) was synthesized according to a literature protocol in H2O (40\u2005mL) was rapidly added 37% formaldehyde at room temperature. The reaction mixture was refluxed for 3\u2005h. After cooling, the resultant white solid was filtered, washed with water, and dried. The crude product of L2\u00b72H2O, (I)2O (1:2 v/v) solution to give colourless crystals suitable for X-ray analysis.Commercially available (Sigma\u2013Aldrich) 1,2-cyclo\u00adhexa\u00adnedi\u00adamine was used as provided. All other chemicals were reagent grade and used without further purification. The starting material, 3,14-dimethyl-2,6,13,17-tetra\u00adaza\u00adtri\u00adcyclo(16.4.0Uiso(H) values of 1.5 and 1.2 Ueq of the parent atoms, respectively. N- and O-bound H atoms were assigned based on a difference Fourier map, and were refined with distance restraints of 0.91\u2005(4) and 0.88\u2005(2)\u2005\u00c5 (using DFIX and DANG commands), respectively, and with Uiso(H) values of 1.2Ueq of the parent atoms.Crystal data, data collection, and structure refinement details are summarized in Table\u00a0210.1107/S2056989017002444/wm5364sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017002444/wm5364Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017002444/wm5364Isup3.cmlSupporting information file. DOI: 1532347CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N6-benzoyl\u00adadenine (BA) and one mol\u00adecule of 4-hy\u00addroxy\u00adbenzoic acid (HBA). The N6-benzoyl\u00adadenine (BA) has an N(7)\u2014H tautomeric form with non-protonated N-1 and N-3 atoms. This tautomeric form is stabilized by a typical intra\u00admolecular N\u2014H\u22efO hydrogen bond on the Hoogsteen face of the purine ring. The primary robust via N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds.The asymmetric unit of the title co-crystal contains one mol\u00adecule of 12H9N5O\u00b7C7H6O3, contains one mol\u00adecule of N6-benzoyl\u00adadenine (BA) and one mol\u00adecule of 4-hy\u00addroxy\u00adbenzoic acid (HBA). The N6-benzoyl\u00adadenine (BA) has an N(7)\u2014H tautomeric form with nonprotonated N-1 and N-3 atoms. This tautomeric form is stabilized by a typical intra\u00admolecular N\u2014H\u22efO hydrogen bond between the carbonyl (C=O) group and the N(7)\u2014H hydrogen on the Hoogsteen face of the purine ring, forming a graph-set S(7) ring motif. The primary robust R22(8) ring motif is formed in the Watson\u2013Crick face via N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds . Weak inter\u00adactions, such as, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 are also observed in this crystal structure.The asymmetric unit of the title co-crystal, C N6-derivatives have plant hormone (kinetin) (Tr). They also offer a variety of hydrogen-bonding donor and acceptor sites mol\u00adecule and one 4-hy\u00addroxy\u00adbenzoic acid (HBA) mol\u00adecule \u2014H tautomeric form with non-protonated N1, N3 and N9 atoms. In the crystal structures of N6-benzoyl\u00adadenine and N6-benzoyl adenine-dl-tartaric acid (1:1) ring motif in the Hoogsteen face. In contrast, it may be noted that in the crystal structure of N6-benzyl\u00adadenine, (where no intra\u00admolecular hydrogen bond is present) the N6-substituent is syn to N1 and distal to N7 and the adenine moiety exists in the N(9)\u2014H tautomeric form le Fig.\u00a01. The bond Table\u00a01 with an via N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds involving the carboxyl group of HBA. The N7 atom is a bifurcated donor and the carbonyl oxygen atom acts as a double acceptor for the N\u2014H\u22efO hydrogen bonds. Inversion-related BA mol\u00adecules form dimers through an array of hydrogen bonds, generating ring motifs, and these dimers are doubly bridged by inversion-related HBA mol\u00adecules supra\u00admolecular ring is formed along the c-axis direction. A weak C8\u2014H8\u22ef\u03c0 inter\u00adaction is also present. Further consolidation of the structure is provided by homo and hetero \u03c0\u2013\u03c0 stacking inter\u00adactions and N6-benzoyl\u00adadenine\u2013dl-tartaric acid (1:1) and 4-hy\u00addroxy\u00adbenzoic acid (35\u2005mg) in an equimolar ratio in a total volume of 30\u2005mL. The mixture was warmed over a water bath for 30\u2005min, filtered, and left aside for a few days. Colourless plate-shaped crystals were collected from the mother solution following slow cooling at room temperature.The title co-crystal was prepared by mixing a hot ethanol solution of Uiso(H) = kUeq, where k = 1.5 for hy\u00addroxy and 1.2 for all other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017002171/hg5481sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017002171/hg5481Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017002171/hg5481Isup3.cmlSupporting information file. DOI: 1531929CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title zwitterion exists in the iminium/phenoxide form. The mol\u00adecule is twisted around the N\u2014C(benzene) bond with the C=N\u2014C\u2014C torsion angle being 39.42\u2005(8)\u00b0. In the crystal, a zigzag supra\u00admolecular chain is sustained by charge-assisted hy\u00addroxy-O\u2014H\u22efO(phenoxide) hydrogen bonding. 17H13NO2 (systematic name: 1-{(1E)-[(4-hy\u00addroxy\u00adphen\u00adyl)iminium\u00adyl]meth\u00adyl}naphthalen-2-olate), features an intra\u00admolecular charge-assisted N+\u2014H\u22efO\u2212 hydrogen bond. A twist in the mol\u00adecule is evident around the N\u2014C(hy\u00addroxy\u00adbenzene) bond [C\u2014N\u2014C\u2014C torsion angle = 39.42\u2005(8)\u00b0] and is reflected in the dihedral angle of 39.42\u2005(8)\u00b0 formed between the aromatic regions of the mol\u00adecule. In the crystal, zigzag supra\u00admolecular chains along the a axis are formed by charge-assisted hy\u00addroxy-O\u2014H\u22efO(phenoxide) hydrogen bonding. These are connected into a layer in the ab plane by charge-assisted hy\u00addroxy\u00adbenzene-C\u2014H\u22efO(phenoxide) inter\u00adactions and \u03c0\u2013\u03c0 contacts [inter-centroid distance between naphthyl-C6 rings = 3.4905\u2005(12)\u2005\u00c5]. Layers stack along the c axis with no specific inter\u00adactions between them. The Hirshfeld surface analysis points to the significance C\u22efH contacts between layers.The title zwitterion, C The dihedral angle between the two aromatic regions is 39.42\u2005(8)\u00b0. The coplanar relationship between the imine and naphthyl residues is stabilized by an intra\u00admolecular charge-assisted N+\u2014H\u22efO\u2212 hydrogen bond, Table\u00a01The mol\u00adecular structure of (I)a axis mediated by hy\u00addroxy-O\u2014H\u22efO(phenoxide) charge-assisted hydrogen bonding, Fig.\u00a02a and Table\u00a01ab plane by charge-assisted hy\u00addroxy\u00adbenzene-C\u2014H\u22efO(phenoxide) inter\u00adactions, Table\u00a01i = 3.4905\u2005(12)\u2005\u00c5 and angle of inclination = 2.68\u2005(8)\u00b0 , Fig.\u00a02b. Layers stack along the c axis with no directional inter\u00adactions between them, Fig.\u00a02c.The most prominent feature of the mol\u00adecular packing is the formation of a zigzag (glide symmetry) supra\u00admolecular chain along the et al., 2017dnorm, Fig.\u00a03The Hirshfeld surface was calculated for (I)a, and those delineated into H\u22efH, C\u22efH/H\u22efC, O\u22efH/H\u22efO and C\u22efC contacts and -(C12\u2013C17) rings through the arrow-shaped distribution with the green points spread about de = di = 1.8\u2005\u00c5. The small contributions from other inter\u00adatomic contacts summarized in Table\u00a03The overall two-dimensional fingerprint plot, Fig.\u00a06et al., 2016i.e. (I)\u00b70.5EtOH and 1.2943\u2005(19)\u2005\u00c5. By contrast to (I)viz. a P21/c form with Z\u2032 = 2 \u00b0 for the C2/c form. Finally, a deprotonated form of (I)et al., 2007The most closely related structure to (I)v/v) by slow evaporation.4-Hy\u00addroxy\u00adaniline was added to a solution of 2-hy\u00addroxy-1-napthaldehyde in ethanol (25\u2005ml). The resulting mixture was refluxed at 333\u2005K and stirred for 2.5\u2005h. The reaction mixture was cooled to room temperature and the resulting orange precipitate was filtered off and washed with cold ethanol to obtain the pure product in 65% yield. Crystals of (I)Uiso(H) values set at 1.2Ueq(C). The O- and N-bound H atoms were located in a difference Fourier map, but were refined with distance restraints of O\u2014H = 0.82\u00b10.01\u2005\u00c5 and N\u2014H = 0.86\u00b10.01\u2005\u00c5, and with Uiso(H) values set at 1.5Ueq(O) and 1.2Ueq(N), respectively. To confirm the positions of the acidic-H atoms, a separate refinement was conducted whereby no distance restraints were applied resulting in O\u2014H and N\u2014H bond lengths of 0.93\u2005(2) and 1.00\u2005(3)\u2005\u00c5, respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S205698901701458X/ex2001sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901701458X/ex2001Isup2.hklStructure factors: contains datablock(s) I. DOI: 1429885CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II centre via the thiol\u00adate S and imine N atoms in each of the two independent mol\u00adecules comprising the asymmetric unit, leading to N2S2 donor sets and distorted tetra\u00adhedal geometries. The crystal features zigzag chains of mol\u00adecules sustained by N\u2014H\u22efN and amine-N\u2014H\u22efS hydrogen bonds.The title thio\u00adsemicarbazonate complex has the ligands coordinating the Zn II complex, [Zn(C18H18N3S)2], (I), features two independent but chemically equivalent mol\u00adecules in the asymmetric unit. In each, the thio\u00adsemicarbazonate monoanion coordinates the ZnII atom via the thiol\u00adate-S and imine-N atoms, with the resulting N2S2 donor set defining a distorted tetra\u00adhedral geometry. The five-membered ZnSCN2 chelate rings adopt distinct conformations in each independent mol\u00adecule, i.e. one ring is almost planar while the other is twisted about the Zn\u2014S bond. In the crystal, the two mol\u00adecules comprising the asymmetric unit are linked by amine-N\u2014H\u22efN(imine) and amine-N\u2014H\u22efS(thiol\u00adate) hydrogen bonds via an eight-membered heterosynthon, {\u22efHNCN\u22efHNCS}. The dimeric aggregates are further consolidated by benzene-C\u2014H\u22efS(thiol\u00adate) inter\u00adactions and are linked into a zigzag supra\u00admolecular chain along the c axis via amine-N\u2014H\u22efS(thiol\u00adate) hydrogen bonds. The chains are connected into a three-dimensional architecture via phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl) and \u03c0\u2013\u03c0 inter\u00adactions, the latter occurring between chelate and phenyl rings [inter-centroid separation = 3.6873\u2005(11)\u2005\u00c5]. The analysis of the Hirshfeld surfaces calculated for (I) emphasizes the different inter\u00adactions formed by the independent mol\u00adecules in the crystal and the impact of the \u03c0\u2013\u03c0 inter\u00adactions between chelate and phenyl rings.The title Zn The similarity in bond lengths extends to the angles subtended at the ZnII atoms which, for the Zn1-containing mol\u00adecule range from 87.00\u2005(5)\u00b0 for S2\u2014Zn1\u2014N6, to 134.00\u2005(5)\u00b0 for S2\u2014Zn1\u2014N3, i.e. a range of 47\u00b0; the acute angle is associated with the chelate angle. A slightly narrower range is noted for the Zn2-containing mol\u00adecule, i.e. 85.99\u2005(5)\u00b0 for S3\u2014Zn2\u2014N9, to 131.29\u2005(5)\u00b0 for S3\u2014Zn2\u2014N12, i.e. about 45\u00b0. The assignment of four-coordinate geometries can be qu\u00adanti\u00adfied by the values of \u03c44, which range from 1.00 for an ideal tetra\u00adhedron to 0.00 for perfect square-planar geometry E, as are the conformations about the ethyl\u00adene bonds, Table\u00a01Two independent mol\u00adecules comprise the asymmetric unit of (I)II atoms to form five-membered ZnSCN2 rings. The chelate rings adopt different conformations in each independent mol\u00adecule. For the Zn1-containing mol\u00adecule, the Zn1/S1/C1/N2/N3 ring is almost planar (r.m.s. deviation = 0.005\u2005\u00c5) but the Zn1/S2/C19/N5/N6 ring is twisted about the Zn1\u2014S2 bond. A similar situation pertains to the Zn2-containing mol\u00adecule where there is a small twist about the Zn2\u2014S3 bond in the Zn2/S3/C37/N8/N9 ring and the Zn2/S4/C55/N11/N12 ring is planar to within an r.m.s. deviation of 0.008\u2005\u00c5. To a first approximation, for each thio\u00adsemicarbazone ligand, all atoms but the terminal ethyl and central phenyl rings lie in a plane. This is qu\u00adanti\u00adfied in the dihedral angle between each five-membered chelate ring and the central and terminal rings of the prop-2-en-1-yl\u00adidene substituent, as summarized in Table\u00a02The thio\u00adsemicarbazone ligands chelate the ZnCrystal Explorer a and Table\u00a04c axis via additional amine-N\u2014H\u22efS(thiol\u00adate) hydrogen-bonds, Fig.\u00a03b. Chains are connected via \u03c0\u2013\u03c0 inter\u00adactions occurring between Zn2-containing mol\u00adecules, involving chelate rings, comprising the Zn2/S4/C55/N11/N12 atoms and phenyl (C61\u2013C66) rings. Precedents for chelate/arene ring inter\u00adactions have been established in the literature involving residues of the Zn1-containing mol\u00adecule exclusively, Table\u00a04c.The most prominent feature of the mol\u00adecular packing is the formation of an eight-membered heterosynthon, {\u22efHNCN\u22efHNCS}, mediated by amine-N\u2014H\u22efN(imine) and amine-N\u2014H\u22efS(thiol\u00adate) hydrogen-bonds which occur between the two mol\u00adecules comprising the asymmetric unit, Fig.\u00a03et al., 2016dnorm in Fig.\u00a04a and e, the bright-red spots near the amine-H1N, H7N, H10N, imime-N2 and thiol\u00adate-S2 and S3 atoms indicate their participation in N\u2014H\u22efN and N\u2014H\u22efS bonds between the two independent mol\u00adecules. In the views of the Hirshfeld surfaces mapped over electrostatic potential for the Zn1-mol\u00adecule in Fig.\u00a04b and c, and for the Zn2-mol\u00adecule in Fig.\u00a04f and g, the hydrogen-bond donors and acceptors are represented by blue and red regions, respectively. Greater insight into inter\u00admolecular inter\u00adactions in the crystal can be obtained by modifying the mapping range for dnorm, as shown in Fig.\u00a04d and h, which reveals additional characteristic spots on the surface. A pair of red spots near amine-HN4 and near phenyl-C7 and C8 in Fig.\u00a04d indicate the presence of short inter-atomic C\u22efH/H\u22efC contacts in the crystal, see Table\u00a05a and e, respectively. The immediate environments about the Zn1- and Zn2-mol\u00adecules within shape-index-mapped Hirshfeld surfaces highlighting hydrogen-bonding and C\u2014H\u22ef\u03c0 inter\u00adactions are illustrated in Fig.\u00a05a and 5b while the C\u2014H\u22ef\u03c0 and their reciprocal, i.e. \u03c0\u22efH\u2014C, contacts involving phenyl-C8 and C32 atoms as donors and phenyl (C31\u2013C36 and C13\u2013C18) rings as acceptors are shown in Fig.\u00a05c.The Hirshfeld surface calculations of (I)i.e. (I)a. In addition, the fingerprint plots delineated into H\u22efH, S\u22efH/H\u22efS, N\u22efH/H\u22efN, C\u22efH/H\u22efC,C\u22efN/N\u22efC and C\u22efC contacts rings of Zn2-mol\u00adecules is evident from the presence of short inter-atomic Zn\u22efC/C\u22efZn contacts, Table\u00a05c, these contacts are reflected by a pair of points with an S-shaped distribution at around de + di \u223c1.9 to 2.1\u2005\u00c5. This \u03c0\u2013\u03c0 stacking is also apparent from the small but effective contributions from Zn\u22efC/C\u22efZn and C\u22efN/N\u22efC contacts to the Hirshfeld surface of the Zn2-mol\u00adecule, Table\u00a06The presence of a short inter-atomic C\u22efS contact between the thiol\u00adate-S3 and phenyl-C66 atoms is evident from the typical H-shaped plot in Fig.\u00a07et al., 2016i.e. of general formula Zn[SC(NHR)=NN=CR\u2032R\u2032\u2032]2 reflecting the inter\u00adest in this class of compound. All of the structures resemble the mol\u00adecular geometry described above for (I)i.e. R\u2032 = R\u2032\u2032 = Me for the R = Ph compound thio\u00adsemi\u00adcarbazide , was used without further purification and was dissolved in heated absolute ethanol (50\u2005ml). Zn(CH3COO)2\u00b72H2O was dissolved separately in heated absolute ethanol (30\u2005ml) and then added into an ethano\u00adlic N-ethyl-N-thio\u00adsemicarbazide solution. The mixture was heated and stirred for about 10 mins, followed by stirring for 1\u2005h at room temperature. The obtained yellow precipitate was filtered, washed with cold ethanol and dried in vacuo. Single crystals were grown at room temperature from the slow evaporation of a solution of di\u00admethyl\u00adformamide and aceto\u00adnitrile (1:1 v/v 20\u2005ml).Analytical grade reagents were used as procured without further purification. Equimolar qu\u00adanti\u00adties of 4-ethyl-3-thio\u00adsemicarbazide and 1,3-di\u00adphenyl\u00adprop-2-en-1-one were dissolved in heated absolute ethanol (30\u2005ml) separately and the mixtures were mixed with stirring. About five drops of concentrated hydro\u00adchloric acid were added to the mixture to catalyse the reaction. The reaction mixture was kept under heating and stirring for about 10 mins, followed by stirring for 1\u2005h at room temperature. The resulting yellow precipitate was filtered off, washed with chilled absolute ethanol and dried Uiso(H) set to 1.2\u20131.5Ueq(C). The nitro\u00adgen-bound H atoms were located in a difference-Fourier map but were refined with a distance restraint of N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S2056989017008064/hb7684sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017008064/hb7684Isup2.hklStructure factors: contains datablock(s) I. DOI: 1553218CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of the centrosymmetric title compound features short N\u2014H\u22ef\u03c0 inter\u00adactions. 8H8N2, is generated by a crystallographic center of symmetry. In the crystal, short N\u2014H\u22ef\u03c0 [H\u22ef\u03c0 = 2.499\u2005(19)\u2005\u00c5] inter\u00adactions link the mol\u00adecules into a herringbone structure.The complete mol\u00adecule of the title compound, C The double bonds C1=C2 and C3=C4 in (I)versus the double-bond length in pyrrole of 1.357\u2005\u00c5. The shortening of the C2\u2014C3 bond and the lengthening of the adjacent C=C double bonds in (I)The complete mol\u00adecule of (I)1 screw axis is 2.499\u2005(19)\u2005\u00c5 \u00c5 Table\u00a01. A surveet al., 2015et al., 1997There are very few examples of similar compounds available in the literature. Most bi\u00adpyrroles have carbonyl groups as substituents on the pyrrole ring in which the N\u2014H group forms hydrogen bonds with the oxygen atom of the carbonyl (Okawara 230\u2005\u00b5l (3.3\u2005mmol) of pyrrole was added to di\u00adchloro\u00admethane (10\u2005ml), degassed and cooled to 195\u2005K. Tri\u00admethyl\u00adsilyl bromide and phenyl iodine trifluoracetic acid were added quickly to the cooled reaction. The mixture was stirred for 1\u2005h then extracted with a saturated sodium bicarbonate solution and purified by column chromatography with a penta\u00adne/ethyl acetate (1:1) solution. Colourless blocks of (I)Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017013433/hb7692sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017013433/hb7692Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017013433/hb7692sup3.pdfSurvey of the Cambridge Database NH-pi bond lengths. DOI: 1575394CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound has a \u2018banana\u2019 shape, with the dihedral angle formed by the outer rings being 8.30\u2005(7)\u00b0. In the crystal, the three-dimensional architecture features nitro\u00adbenzene-C\u2014H\u22efO(carbon\u00adyl), pyrrole-C\u2014H\u22efO(nitro), \u03c0(pyrrole)\u2014\u03c0(nitro\u00adbenzene) and nitro-O\u22ef\u03c0(pyrrole) inter\u00adactions. 12H12N2O4, the di\u00adhydro\u00adpyrrole ring is almost planar (r.m.s. deviation = 0.0049\u2005\u00c5) and is nearly coplanar with the adjacent C2O2 residue [dihedral angle = 4.56\u2005(9)\u00b0], which links to the 4-nitro\u00adbenzene substituent [dihedral angle = 4.58\u2005(8)\u00b0]. The mol\u00adecule is concave, with the outer rings lying to the same side of the central C2O2 residue and being inclined to each other [dihedral angle = 8.30\u2005(7)\u00b0]. In the crystal, supra\u00admolecular layers parallel to (10-5) are sustained by nitro\u00adbenzene-C\u2014H\u22efO(carbon\u00adyl) and pyrrole-C\u2014H\u22efO(nitro) inter\u00adactions. The layers are connected into a three-dimensional architecture by \u03c0(pyrrole)\u2013\u03c0(nitro\u00adbenzene) stacking [inter-centroid separation = 3.7414\u2005(10)\u2005\u00c5] and nitro-O\u22ef\u03c0(pyrrole) inter\u00adactions.In the title compound, C The planarity extends to the 4-nitro\u00adbenzene ring, with the dihedral angle between the C2O2 and C6 planes being 4.58\u2005(8)\u00b0. However, the mol\u00adecule is not planar but rather is curved as the outer rings lie to the same side of the central C2O2 residue; the dihedral angle = 8.30\u2005(7)\u00b0. To a first approximation, the nitro group is co-planar with the benzene ring to which is connected, as seen in the value of the O4\u2014N2\u2014C10\u2014C9 torsion angle of 173.50\u2005(15)\u00b0.The mol\u00adecular structure of (I)3OCO}2 synthons. The dimeric aggregates are connected into a supra\u00admolecular layer via pyrrole-C4\u2014H\u22efO3(nitro) inter\u00adactions. The layers lie parallel to \u2013\u03c0(C7\u2013C12)i stacking inter\u00adactions occur between pyrrole and nitro\u00adbenzene rings: inter-centroid separation = 3.7414\u2005(10)\u2005\u00c5 and angle of inclination = 7.99\u2005(9)\u00b0 for symmetry code: (i): x, \u2212y, z. The other inter\u00adactions between layers are of the type nitro-O4\u22ef\u03c0, Table\u00a01et al., 2008b.The mol\u00adecular packing of (I)et al., 2017The Hirshfeld surface calculations for (I)dnorm in Fig.\u00a03B and ester-C5 atoms, respectively, Fig.\u00a04a. The other short inter\u00adatomic C\u22efH/H\u22efC contacts between the nitro\u00adbenzene-H11 and pyrrole-C2 and C3 atoms contacts is highlighted in Fig.\u00a04b, showing the Hirshfeld surface mapped over the electrostatic potential.In addition to the bright-red spots on the Hirshfeld surface mapped over s Table\u00a02 are intret al., 2007a\u2013d, respectively, and the percentage contribution from the identified inter\u00adatomic contacts to the Hirshfeld surface are summarized in Table\u00a03i.e. 39.0%, contribution from H\u22efH contacts to the overall surface is due to the involvement of many hydrogen atoms in directional inter\u00admolecular inter\u00adactions, e.g. C\u2014H\u22efO, \u03c0 (Tables 1b). Conversely, the relatively significant contribution of 33.8% from O\u22efH/H\u22efO contacts to the Hirshfeld surface is consistent with this observation. The fingerprint plot delineated into O\u22efH/H\u22efO contacts features a pair of green aligned points within the pair of spikes with their tips at de + di \u223c2.3\u2005\u00c5 superimposed upon a distribution blue points characterizing inter\u00admolecular C\u2014H\u22efO inter\u00adactions. The short inter\u00adatomic C\u22efH/H\u22efC contacts in the inter- and intra-layer regions are represented by the two pairs of short forceps-like spikes at de + di \u223c2.8 and 2.9\u2005\u00c5, respectively, in Fig.\u00a05d. The small but discernible contributions from inter\u00adatomic C\u22efC and C\u22efN/N\u22efC contacts 2 Di\u00adhydro\u00adpyrrole rings as found in (I)M solution, 10.56\u2005mmol) in dry toluene was then added. The bath was removed and the solution stirred for 2\u2005h at room temperature. Subsequently, the flask was immersed for 20\u2005min in an oil bath preheated to 393\u2013403\u2005K with a reflux condenser. The solution was concentrated in a rotary evaporator and the residue was purified by flash column chromatography on silica gel, using a mixture of EtOAc/n-hexane (1:4) as the eluent. The yield of (I)2Cl2 solution.A solution of (4-nitro\u00adphen\u00adyl)methyl 2-hy\u00addroxy\u00adpyrrolidine-1-carboxyl\u00adate in toluene (100\u2005ml) was cooled to 273\u2005K in an ice/water bath. Under an atmosphere of nitro\u00adgen, 2,4-lutidine was added to this solution. The solution was stirred for 15\u2005min at 273\u2005K. A tri\u00adfluoro\u00adacetic anhydride (TFAA) solution : \u03b4 8.21 , 7.54 , 6.80 and 6.68 , 5.35 , 5.03 , 3.71 , 2.46 . 13C NMR : \u03b4 = 152.3 (CO2R), 151.5 (CO2R), 147.8 (C4\u2032), 144.9 (C1\u2032), 129.8 (C2), 129.2 (C2), 128.4 (C2\u2032 and C6\u2032), 128.3 (C2\u2032 and C6\u2032), 123.9 (C3\u2032 and C5\u2032), 109.4 (C3), 65.8 (CH2), 65.6 (CH2), 45.8 (C5), 45.4 (C5), 30.1 (C4), 29.0 (C4). ESI\u2013MS (m/z) calculated for C12H12N2O4 248.07971, found 248.07876.Spectroscopic characterization. Uiso(H) set to 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989018002451/hb7736sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018002451/hb7736Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018002451/hb7736Isup3.cmlSupporting information file. DOI: 1823263CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the 1:2 co-crystalline adducts are linked by \u03c0\u2013\u03c0 stacking inter\u00adactions. 3,8]undecano (TATU) with 4-chloro-3,5-di\u00admethyl\u00adphenol led to the formation of the title co-crystal, C7H14N4\u00b72C8H9ClO. The asymmetric unit contains one aminal cage mol\u00adecule and two phenol mol\u00adecules linked via two O\u2014H\u22efN hydrogen bonds. In the aminal cage, the N\u2013CH2\u2013CH2\u2013N unit is slightly distorted from a syn periplanar geometry. Aromatic \u03c0\u2013\u03c0 stacking between the benzene rings from two different neighbouring phenol mol\u00adecules [centroid\u2013centroid distance = 4.0570\u2005(11)\u2005\u00c5] consolidates the crystal packing.Solvent-free treatment of 1,3,6,8-tetra\u00adaza\u00adtri\u00adcyclo\u00ad[4.3.1.1 Nitro\u00adgCg8\u22efCg9i = 4.0570\u2005(11); symmetry code: (i) x, y, Cgz; 8 and Cg9 are the centroids of the C11\u201316 and C21\u2013C26 rings, respectively] and the benzene ring planes are inclined to one another by 0.58\u2005(9)\u00b0. These additional contacts link the three-membered co-crystal units into chains approximately parallel to undecane hydro\u00adquinone and 4-chloro-3,5-di\u00admethyl\u00adphenol was ground using a mortar and pestle at room temperature for 15\u2005min. Completion of the reaction was monitored by TLC. The mixture was recrystallized from n-hexane solution to obtain colourless crystals suitable for X-ray analysis, m.p. = 375\u2013376\u2005K. (yield: 63%).A mixture of 1,3,6,8-tetra\u00adaza\u00adtri\u00adcyclo\u00ad[4.3.1.1Uiso(H) set to 1.2Ueq of the parent atom. The hydroxyl H atoms were refined freely.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016016650/sj5512sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016016650/sj5512Isup2.hklStructure factors: contains datablock(s) I. DOI: 1510356CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II cation is five-coordinated in a distorted trigonal\u2013bipyramidal geometry. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (58.4%), H\u22efC/C\u22efH (20.3%) and H\u22efO/O\u22efH (18.3%) inter\u00adactions.In the title Zn complex, the Zn 10H11O2)2(C6H6N2O)2(H2O)], contains one half of the complex mol\u00adecule, and the ZnII cation and the water O atom lie on a twofold rotation axis. The ZnII cation is coordinated by two carboxyl\u00adate O atoms of the two symmetry-related 2,4,6-tri\u00admethyl\u00adbenzoate (TMB) anions and by the water O atom at distances of 2.0311\u2005(16) and 2.076\u2005(2)\u2005\u00c5 to form a slightly distorted trigonal\u2013planar arrangement, while the distorted trigonal\u2013bipyramidal coordination sphere is completed by the two pyridine N atoms of the two symmetry-related monodentate nicotinamide (NA) ligands at distances of 2.2066\u2005(19)\u2005\u00c5 in the axial positions. In the crystal, mol\u00adecules are linked via inter\u00admolecular N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds with R22(12), R33(10) and R33(16) ring motifs, forming a double-column structure running along the c-axis direction. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (58.4%), H\u22efC/C\u22efH (20.3%) and H\u22efO/O\u22efH (18.3%) inter\u00adactions.The asymmetric unit of the title complex, [Zn(C A deficiency of this vitamin leads to loss of copper from the body, known as pellagra disease. Victims of pellagra show unusually high serum and urinary copper levels 2(C6H6N2O)2] \u00b7H2O (I)The transition metal complexes with ligands of biochemical inter\u00adest as imidazole and some II cation (site symmetry 2), one 2,4,6-tri\u00admethyl\u00adbenzoate (TMB) anion and one nicotinamide (NA) mol\u00adecule together with one water mol\u00adecule (point group symmetry 2), all ligands coordinating in a monodentate manner 1\u00a0\u2212\u00a0x, y, z] of the two symmetry-related monodentate TMB anions and the coordinating water O atom (O4) are at distances of 2.0311\u2005(16) and 2.076\u2005(2)\u2005\u00c5, respectively, around the Zn1 atom and form a slightly distorted triangular planar arrangement. The sum of the bond angles O2\u2014Zn1\u2014O2i [95.38\u2005(9)\u00b0], O2\u2014Zn1\u2014O4 [132.31\u2005(5)\u00b0] and O2\u2014Zn1\u2014O4i [132.31\u2005(5)\u00b0] in the basal plane around ZnII cation is 360\u00b0. This confirms the presence of the ZnII cation with very slight deviation from the basal plane. The slightly distorted trigonal\u2013bipyramidal coordination sphere is completed by the two pyridine N atoms (N1 and N1i) of the two symmetry-related monodentate NA ligands at distances of 2.2066\u2005(19)\u2005\u00c5 in the axial positions. The index of trigonality \u03c4 and C1\u2014O2 [1.259\u2005(3)\u2005\u00c5] bonds in the carboxyl\u00adate groups indicate delocalized bonding arrangements rather than localized single and double bonds. The O2\u2014C1\u2014O1 bond angle [121.8\u2005(2)\u00b0] seems to be slightly decreased than that present in a free acid [122.2\u00b0], in which the O2\u2014C1\u2014O1 bond angle may be compared with the corresponding values of 123.5\u2005(2) and 120.4\u2005(2)\u00b0 in (II), 125.2\u2005(5)\u00b0 in (III), 119.2\u2005(3) and 123.8\u2005(2)\u00b0 in (IV), 123.6\u2005(3) and 119.4\u2005(3)\u00b0 in (V), 124.4\u2005(2)\u00b0 in (VI) and 124.3\u2005(2)\u00b0 in (VII), where the benzoate ions coordinate to the metal atoms only monodentately in (III), (VI) and (VII), and both monodentately and bidentately in (II), (IV) and (V). The Zn1 atom lies 0.0817\u2005(1)\u2005\u00c5 above of the planar (O1/O2/C1) carboxyl\u00adate group. In the TMB anion, the carboxyl\u00adate group is twisted away from the attached benzene C2\u2013C7 ring by 61.32\u2005(14)\u00b0, while the benzene ring and the pyridine N1/C11\u2013C15 ring are oriented at a dihedral angle of 81.90\u2005(8)\u00b0.2 group links to the non-coordinating carboxyl\u00adate and NA oxygen atoms via inter\u00admolecular N\u2014H\u22efO hydrogen bonds, and the water mol\u00adecule links to the NA oxygen atoms via inter\u00admolecular O\u2014H\u22efO hydrogen bonds , the 18.3% contribution to the HS arises from the inter\u00admolecular O\u2014H\u22efO hydrogen bonding and is viewed as pair of spikes with the tip at de + di \u223c1.9\u2005\u00c5. The short H\u22efO/O\u22efH contacts are masked by strong O\u2014H\u22efO hydrogen bonding in this plot. The H\u22efN/N\u22efH contacts in the structure with 1.9% contribution to the HS has a symmetrical distribution of points with the tips at de + di \u223c2.8\u2005\u00c5 arising from the short inter\u00adatomic H\u22efN/N\u22efH contact . The HSs mapped over shape-index, curvedness and those with the function dnorm plotted onto the surface are shown for the H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH and H\u22efN/N\u22efH inter\u00adactions are shown in Figs. s1\u2013s3 in the Supporting Information.A Hirshfeld surface (HS) analysis in H2O (50\u2005ml) and nicotinamide in H2O (25\u2005ml) with sodium 2,4,6-tri\u00admethyl\u00adbenzoate in H2O (150\u2005ml) at room temperature. The mixture was set aside to crystallize at ambient temperature for ten weeks, giving colourless single crystals . FT\u2013IR: 3396, 3111, 2953, 2919, 2740, 2321, 1947, 1693, 1665, 1621, 1601, 1584, 1445, 1397, 1199, 1113, 1047, 860, 839, 797, 731, 647, 614, 545, 559\u2005cm\u22121.The title compound was prepared by the reaction of ZnSO2 group were also located in a difference Fourier map and the positions were refined with Uiso(H) = 1.5Ueq(N). The C-bound H atoms were positioned geometrically with C\u2014H = 0.93 and 0.96\u2005\u00c5 for aromatic and methyl H-atoms, respectively, and refined as riding with Uiso(H) = k \u00d7 Ueq(C), where k = 1.5 for methyl H-atoms and k = 1.2 for aromatic H-atoms.The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0210.1107/S2056989017011690/is5478sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017011690/is5478Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017011690/is5478sup3.pdfHirsfeld surface plotted over shape-index. DOI: 10.1107/S2056989017011690/is5478sup4.pdfHirshfeld surface plotted over curvedness. DOI: 10.1107/S2056989017011690/is5478sup5.pdfHirshfeld surfaces plotted over dnorm. DOI: 1567723CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the isomeric title compounds, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bond, and C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions build different supra\u00admolecular architectures. 14H13Cl2NO2S, namely 3,5-di\u00adchloro-N--benzene\u00adsulfonamide (I), 3,5-di\u00adchloro-N-benzene\u00adsulfonamide (II) and 3,5-di\u00adchloro-N-benzene\u00adsulfonamide (III) are described. The mol\u00adecules of all the three compounds are U-shaped with the two aromatic rings inclined at 41.3\u2005(6)\u00b0 in (I), 42.1\u2005(2)\u00b0 in (II) and 54.4\u2005(3)\u00b0 in (III). The mol\u00adecular conformation of (II) is stabilized by intra\u00admolecular C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions. The crystal structure of (I) features N\u2014H\u22efO hydrogen-bonded R22(8) loops inter\u00adconnected via C(7) chains of C\u2014H\u22efO inter\u00adactions, forming a three-dimensional architecture. The structure also features \u03c0\u2013\u03c0 inter\u00adactions [Cg\u22efCg = 3.6970\u2005(14)\u2005\u00c5]. In (II), N\u2014H\u22efO hydrogen-bonded R22(8) loops are inter\u00adconnected via \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.606\u2005(3)\u2005\u00c5] to form a one-dimensional architecture running parallel to the a axis. In (III), adjacent C(4) chains of N\u2014H\u22efO hydrogen-bonded mol\u00adecules running parallel to [010] are connected via C\u2014H\u22ef\u03c0 inter\u00adactions, forming sheets parallel to the ab plane. Neighbouring sheets are linked via offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.8303\u2005(16)\u2005\u00c5] to form a three-dimensional architecture.The crystal structures of three isomeric compounds of formula C The structure also features \u03c0\u2013\u03c0 inter\u00adactions involving the benzene\u00adsulfonyl ring and the aniline ring as illustrated in Fig.\u00a04Cg1\u22efCg2iii = 3.6970\u2005(14)\u2005\u00c5; Cg1 and Cg2 are the centroids of the C1\u2013C6 and C7\u2013C12 rings, respectively; symmetry code: (iii) x, \u2212y, z].The crystal structure of (I)ps Fig.\u00a04. The (8)i hydrogen-bonded via \u03c0\u2013\u03c0 inter\u00adactions involving inversion-related benzene\u00adsulfonyl rings, forming a one-dimensional architecture running parallel to the a axis, as shown in Fig.\u00a05Cg1\u22efCg1ii = 3.606\u2005(3)\u2005\u00c5; Cg1 is the centroid of the C1\u2013C6 ring; symmetry code: (ii) 2\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, \u2212z].In (II)ps Fig.\u00a05 are connvia N1\u2014H1\u22efO1i hydrogen bonds chains running parallel to [010]. Adjacent chains are connected by C14\u2014H14B\u22ef\u03c0 inter\u00adactions involving the aniline ring, forming two-dimensional sheets parallel to the ab plane. Neighbouring sheets are further linked via offset \u03c0\u2013\u03c0 inter\u00adactions involving inversion-related benzene\u00adsulfonyl rings, forming a three dimensional architecture as as illustrated in Fig.\u00a07Cg1\u22efCg1i = 3.8303\u2005(16)\u2005\u00c5, inter\u00adplanar distance = 3.3874\u2005(11)\u2005\u00c5, slippage 1.788\u2005(3)\u2005\u00c5; Cg1 is the centroid of the C1\u2013C6 ring; symmetry code: (iii) 1\u00a0\u2212\u00a0x, \u2212y, \u2212z].In the crystal structure of (III)ds Fig.\u00a06 to form N-(substitutedphen\u00adyl)-benzene\u00adsulfon\u00adamides, namely 3,5-di\u00adchloro-N-(4-methyl\u00adphen\u00adyl)benzene\u00adsulfonamide [Shakuntala, Naveen et al., 2017N-benzene\u00adsulfonamide [Shak\u00aduntala, Lokanath et al., 2017via N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, whereas in (V) the three-dimensional supra\u00admolecular architecture is built through N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, Cl\u22efCl contacts and \u03c0\u2013\u03c0 inter\u00adactions.Two 3,5-di\u00adchloro-et al., 2015The title compounds were prepared according to a literature method (Rodrigues Uiso(H) = 1.2 or 1.5Ueq(C). A rotating model was applied to the methyl groups. To improve considerably the values of R1, wR2, and S (goodness-of-fit), a low-angle reflection partially obscured by the beam-stop (100) was omitted from the final refinement of (III)Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989017005230/rz5211sup1.cifCrystal structure: contains datablock(s) I, II, III, shelx. DOI: 10.1107/S2056989017005230/rz5211Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017005230/rz5211IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989017005230/rz5211IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989017005230/rz5211Isup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989017005230/rz5211IIsup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989017005230/rz5211IIIsup7.cmlSupporting information file. DOI: 1542706, 1542704, 1542705CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "C(6) helical chain running along the [100] direction.In the two independent mol\u00adecules in the asymmetric unit of the title compound, the cyclo\u00adhexane rings adopt a chair conformation, while the oxane rings are also puckered. In the crystal, O\u2014H\u22ef O hydrogen bonds connect adjacent mol\u00adecules, forming a 20H34O2, contains two crystallographically independent mol\u00adecules (1 and 2) with similar conformations. In both mol\u00adecules, the cyclo\u00adhexane rings adopt a chair conformation, while the oxane rings are also puckered. In the crystal, O\u2014H\u22efO hydrogen bonds connect adjacent mol\u00adecules, forming C(6) helical chains located around a 21 screw axis and running along the crystallographic a axis. The packing of these chains is governed only by van der Waals inter\u00adactions. Semi-empirical PM3 quantum chemical calculations are in a satisfactory agreement with the structural results of the X-ray structure analysis. The absolute structure was indeterminate in the present experiment.The asymmetric unit of the title compound, C Sideritis genus belonging to the Lamiaceae family is represented by more than 150 species in the world \u2005\u00c5, \u03b8 = 0.0\u2005(3), \u03c6 = 270\u2005(81)\u00b0 and QT = 0.584\u2005(3)\u2005\u00c5, \u03b8 = 4.4\u2005(3), \u03c6 = 59\u2005(4)\u00b0, respectively. The oxane ring (O2/C11/C12/C15\u2013C17) is also puckered, with puckering parameters QT = 0.551\u2005(3)\u2005\u00c5, \u03b8 = 12.1\u2005(3) and \u03c6 = 133.5\u2005(16)\u00b0. The equivalent rings in mol\u00adecule 2 have as puckering parameters QT = 0.534\u2005(3)\u2005\u00c5, \u03b8 = 1.9\u2005(3), \u03c6 = 296\u2005(11)\u00b0, QT = 0.583\u2005(3)\u2005\u00c5, \u03b8 = 5.0\u2005(3), \u03c6 = 72\u2005(3)\u00b0 and QT = 0.554\u2005(3)\u2005\u00c5, \u03b8 = 11.9\u2005(3), \u03c6 = 127.2\u2005(15)\u00b0, respectively. Bond lengths and angles are within normal range, comparable with each other and with those reported for similar structures in the literature helical chains located around a 21 screw axis running along the crystallographic a axis is a semi-empirical method for the quantum calculation of the mol\u00adecular electronic structure in computational chemistry. It is based on the neglect of differential diatomic overlap integral approximation. The semi-empirical PM3 parameterization used in the MOPAC program is widely used to derive charges, dipole moments and bond lengths. The computed quantum chemical descriptors include bond lengths, bond angles, torsion angles, atom charges, HOMO and LUMO energy levels, dipole moment, polarizability, etc. In the present case, the geometry of the mol\u00adecule of the title compound was calculated with a semi-empirical PM3 method , with the sole exception of the angles in the meth\u00adoxy groups. This may be ascribed to the steric inter\u00adactions between adjacent mol\u00adecules in the crystal structure.\u22121 flow rate. 16 fractions, each of which was 150\u2005mL, were collected. Similar fractions were combined according to the TLC profile. Further purification was carried out with silica gel column chromatography to isolate the title compound. Colourless prisms were recrystallized from ethanol solution.The aerial part of the plant material (5\u2005g) was extracted with ethyl acetate (3 \u00d7 20\u2005mL). After removal of the solvent by rotary evaporator, the extract was subjected to column chromatography (2.5 \u00d7 70\u2005cm); sephadex LH-20 (50\u2005g) was used as a stationary phase and methanol was used as a mobile phase with a 0.25\u2005ml\u2005minUiso fixed at 1.5 times Ueq(O). H atoms bound to carbon were positioned geometrically and allowed to ride on their parent atoms with Uiso = 1.2Ueq(C) and with Uiso = 1.5Ueq(C) (C\u2014H = 0.96\u2005\u00c5) for methyl H atoms. The absolute structure was indeterminate in the present experiment.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016013864/bg2593sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989016013864/bg2593Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016013864/bg2593sup3.pdfSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016013864/bg2593sup4.tifSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016013864/bg2593Isup5.cmlSupporting information file. DOI: 1501445CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecular and crystal structure of the title imidazole derivative is reported. The structure features an extensive O\u2014H\u22efN, C\u2014H\u22efO/N and C\u2014H\u22ef\u03c0(ring) hydrogen-bonding network. 24H21N3O3, crystallizes with two unique but closely r.m.s. overlay fit = 0.215\u2005\u00c5) comparable mol\u00adecules (1 and 2) in the asymmetric unit of the triclinic unit cell. In molecule 1, the dihedral angles between the central imidazlole ring and the benzene-ring substituents are 42.51\u2005(9), 45.41\u2005(9) and 56.92\u2005(8)\u00b0, respectively. Comparable data for molecule 2 are 39.36\u2005(10), 34.45\u2005(11) and 60.34\u2005(8)\u00b0, respectively. The rings at the 2-positions carry p-nitro substituents that subtend dihedral angles of 12.9\u2005(4)\u00b0 in mol\u00adecule 1 and 11.7\u2005(4)\u00b0 in mol\u00adecule 2 to their respective benzene ring planes. The imidazole rings also have propan-2-ol substituents on the 1-N atoms, which adopt extended conformations for the N\u2014C\u2014C\u2014C chains. In the crystal, classical O\u2014H\u22efN hydrogen bonds combine with C\u2014H\u22efO, C\u2014H\u22efN and C\u2014H\u22ef\u03c0(ring) hydrogen bonds and stack the molecules along the a-axis direction.The title compound, C The benzene rings are inclined to the imidazole ring plane at angles of 42.51\u2005(9) and 39.36\u2005(10)\u00b0 for C121\u2013C126 and C221\u2013C226, 45.41\u2005(9) and 34.45\u2005(11)\u00b0 for C141\u2013C146 and C241\u2013C246 and 56.92\u2005(8) and 60.34\u2005(8) for C151\u2013C156 and C251\u2013C256, values that further attest to the close similarities between the structures of the two unique mol\u00adecules. Bond lengths and angles in the two mol\u00adecules are also similar and compare well with those found in comparable mol\u00adecules with iso\u00adpropanol substituents at the 1-position chains along the b-axis direction, Fig.\u00a03c, in this case head-to-head, due to C153\u2014H153\u22efCg5 and C255\u2014H255\u22efCg1 contacts combined with C152\u2014H152\u22efO212, C153\u2014H153\u22efN21 and C256\u2014H256\u22efO112 hydrogen bonds, Fig.\u00a04via C243\u2014H243\u22efO23 hydrogen bonds, forming C(13) chains along a, Fig.\u00a05a. Overall, these numerous contacts generate layers of molecules of (I)a-axis direction, Fig.\u00a06In the crystal, classical O112\u2014H11et al., 20162 substituent on N1 are reasonably common with 43 entries. However, restricting the search to compounds with iso\u00adpropanol substituents on N1 reduces the hits to three reports of our work to produce compounds with 4-benzoic acid . All other atoms were refined using a riding model with d(C\u2014H) = 0.95\u2005\u00c5 for aromatic, 1.00\u2005\u00c5 for methine and 0.98\u2005\u00c5 for CH2 atoms, all with Uiso(H) = 1.2Ueq(C). For methyl H atoms d(C\u2014H) = 0.98\u2005\u00c5 and Uiso(H) = 1.5Ueq(C). One low angle reflection with Fo << Fc that may have been affected by the beamstop was omitted from the final refinement cycles.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017011057/hg5491sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017011057/hg5491Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017011057/hg5491Isup3.cmlSupporting information file. DOI: 1565059CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I atom in the title compound is two-coordinate, being bound to two pyridine N atoms from two N-(pyridin-3-ylmeth\u00adyl)pyridin-3-amine ligands in a slightly distorted linear fashion. Each AgI ion bridges the dipyridyl-type ligands to form polymeric zigzag chains. The chains are connected via Ag\u22efAg and \u03c0\u2013\u03c0 inter\u00adactions, forming a corrugated layer parallel to ]PF6\u00b72CH3CN or {[AgL]\u00b7PF6\u00b72CH3CN}n, L = N-(pyridin-3-ylmeth\u00adyl)pyridin-3-amine, comprises one AgI atom, one L ligand, two aceto\u00adnitrile solvent mol\u00adecules and one PF6\u2212 anion disordered over two orientations in a 0.567\u2005(11):0.433\u2005(11) ratio. Each AgI atom is coordinated by two pyridine N atoms from two L ligands in a slightly distorted linear coordination geometry [N\u2014Ag\u2014N = 170.55\u2005(8)\u00b0]. Each L ligand bridges two AgI ions, resulting in the formation of a zigzag chain propagating along the [101] direction. In the crystal, Ag\u22efAg contacts [3.3023\u2005(5)\u2005\u00c5] and inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid-to-centroid distance = 3.5922\u2005(15)\u2005\u00c5] between the pyridine rings link these chains into a corrugated layer parallel to the \u2005\u00c5] between the layer and the inter\u00adcalated guests and between the inter\u00adcalated guests, forming a three-dimensional supra\u00admolecular network.The asymmetric unit in the title compound, [Ag(C The silver(I) atom is coordinated by two pyridine N atoms (N1 and N2) from two symmetry-related L ligands, leading to the formation of an infinite zigzag chain propagating along the [101] direction. Thus, the AgI atom is two-coordinated in a slightly distorted linear coordination geometry . All other H atoms were positioned geometrically and refined as riding: C\u2014H = 0.95\u2005\u00c5 for Csp2\u2014H, 0.99\u2005\u00c5 for methyl\u00adene C\u2014H and 0.98\u2005\u00c5 for methyl C\u2014H with Uiso(H) = 1.5Ueq (C-meth\u00adyl) and 1.2Ueq(C) for other C-bound H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017013421/su5393sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989017013421/su5393Isup2.hklStructure factors: contains datablock(s) I. DOI: 1575393CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The di\u00adthio\u00adcarbamate ligand coordinates to the metal atom in an asymmetric mode with the resulting C2S4 donor set defining a skew trapezoidal bipyramidal geometry; the n-butyl groups are disposed to lie over the longer Sn\u2014S bonds. Supra\u00admolecular chains aligned along the a-axis direction and sustained by methyl\u00adene-C\u2014H\u22efS(weakly coordinating) inter\u00adactions feature in the mol\u00adecular packing. A Hirshfeld surface analysis reveals the dominance of H\u22efH contacts in the crystal.The complete mol\u00adecule of the title compound, [Sn(C R2Sn(S2CNRR\u2032)2 is diverse with coordination geometries ranging from five, as in trigonal bipyramid (t-Bu)2Sn(S2CNMe2)2 2 Me]2, with a di\u00adthio\u00adcarbamate ligand with dissimilar substituents, was found to be octa\u00adhedral but, Ph2Sn[S2CN(CH2CH2OMe)2]2, with the di\u00adthio\u00adcarbamate ligand having similar substituents, was skew trapezoidal bipyramidal Me]2, (I)via an analysis of the Hirshfeld surface.The structural chemistry of mol\u00adecules with the general formula n-butyl groups lying on the plane, Fig.\u00a01i.e. the difference between the Sn\u2014Slong and Sn\u2014Sshort distances, being ca 0.39\u2005\u00c5, Table\u00a01n-butyl groups. The four S atoms are co-planar and define a skewed trapezoidal plane, and the \u03b1-C atoms are disposed over the weaker Sn\u2014S bonds so that the C2S4 donor set defines a skew trapezoidal bipyramidal geometry.The asymmetric unit of (I)a-axis direction, Fig.\u00a02a and Table\u00a02b. In order to ascertain more information of the nature of inter\u00adactions between mol\u00adecules, the mol\u00adecular packing and its Hirshfeld surface was analysed, as discussed in Hirshfeld surface analysis.The only notable contacts identified in the mol\u00adecular packing are methyl\u00adene-C\u2014H\u22efS(weakly coordinating) inter\u00adactions that assemble mol\u00adecules into linear supra\u00admolecular chains propagating along the et al., 2016dnorm, in the range \u22120.298 to +1.346 au, in Fig.\u00a03C and H13B of the disordered methyl groups, i.e. deviating from mirror symmetry, indicate their participation in specific inter\u00admolecular H\u22efH inter\u00adactions. In the crystal, these lead to a supra\u00admolecular chain along the c axis. The presence of this di\u00adhydrogen inter\u00adaction, resulting from disparate charges on respective hydrogen atoms, can also be viewed by the different curvatures and electrostatic potentials around these atoms on the Hirshfeld surface mapped over the electrostatic potential in the range \u22120.082 to +0.163 au, Fig.\u00a04dnorm, highlighting the inter\u00admolecular C\u2014H\u22efS and H\u22efH inter\u00adactions.The Hirshfeld surface analysis for (I)a, and those delineated region, Fig.\u00a06a and b, show the dominance of these inter\u00adactions in the mol\u00adecular packing. The peak in the plot corresponding to a second short inter\u00adatomic H\u22efH contact, i.e. between methyl-H2B and methyl\u00adene-H10A, Table\u00a04B\u22efH13B inter\u00adaction. The di\u00adhydrogen H\u22efH bonding also results in short inter\u00adatomic C\u22efH/H\u22efC contacts, Table\u00a04de + di \u223c 2.8\u2005\u00c5 in the delineated fingerprint plot, Fig.\u00a06c; the other inter\u00adatomic short C\u22efH/H\u22efC contact is merged within the plot. The presence of the weak C\u2014H\u22efS inter\u00adactions, Table\u00a02d, and is evident as a pair of broad peaks at de + di \u223c 2.9\u2005\u00c5. The fingerprint plots delineated into O\u22efH/H\u22efO and N\u22efH/H\u22efN contacts, Fig.\u00a06e and f, contribute in a minor fashion to the Hirshfeld surface and their characteristic points are longer than their respective van der Waals separations, i.e. longer than 2.72 and 2.75\u2005\u00c5, respectively, and hence it is likely they do not make any significant contribution to the mol\u00adecular packing.In the fingerprint plot delineated into H\u22efH contacts, Fig.\u00a06b) that that H atoms adopt positions as far apart from each other as possible rather than participate in \u2018non-bonded steric repulsion\u2019 et al., 2016n-Bu2Sn(S2CNRR\u2019)2. One structure, i.e. R = R\u2032 = i-Pr , seven, i.e. R = Me, R\u2032 = n-Bu Me2 piperazine 2.The inter\u00adest in organotin di\u00adthio\u00adcarbamates is reflected in the relatively large number of crystal structures available in the crystallographic literature (Groom n-Bu2Sn(S2CNRR\u2032)2 is that they all conform to the same structural motif as adopted for (I)short bond lengths in these structures span a relatively narrow range of 2.51 to 2.55\u2005\u00c5 and cluster around 2.53\u2005\u00c5. As might be anti\u00adcipated, a wider range is exhibited by the Sn\u2014Slong bonds, i.e. 2.83 to 3.08\u2005\u00c5 and these cluster around 2.96\u2005\u00c5. Given the range of Sn\u2014Sshort bond lengths is 0.04\u2005\u00c5 and that for Sn\u2014Slong is 0.25\u2005\u00c5, the observation that differences between the average values of Sn\u2014Sshort and Sn\u2014Slong span a range of 0.43\u2005\u00c5 indicates no specific correlations exist between Sn\u2014Sshort and Sn\u2014Slong bond lengths. The Sshort\u2014Sn\u2014Sshort, Slong\u2014Sn\u2014Slong and C\u2014Sn\u2014C angles cluster around 83, 147 and 136\u00b0, respectively. However, these angles span ranges of 8\u00b0 (range: 80 to 88\u00b0), 10\u00b0 (140 to 151\u00b0) and 18\u00b0 (127 to 145\u00b0), respectively. The disparity in the S\u2014Sn\u2014S angles is as expected from the adopted coordination geometry. While, generally, the Slong\u2014Sn\u2014Slong angles are wider than the C\u2014Sn\u2014C angles, there are three exceptional structures, namely R = R\u2032 = Et 2 structural motif does not translate to the diphenyl analogues, i.e. Ph2Sn(S2CNRR\u2032)2. Of the 19 structures conforming to this general formula, seven resemble the skew trapezoidal bipyramidal motif with the majority, i.e. twelve, having a cis-disposition of the tin-bound phenyl substituents. In this context, it is noteworthy that all structures of the general formula Sn(S2CNRR\u2032)2X2, where X = halide, are invariably cis-S4X2 octa\u00adhedral 2 compounds.The homogeneity in the ca 277\u2005K) for 30\u2005min. 25% Ammonia solution (ca 2\u2005ml) was added to make the solution basic. Then, a cold ethanol solution of carbon di\u00adsulfide (10\u2005mmol) was added to the solution followed by stirring for about 2\u2005h. Next, di-n-butyl\u00adtin(IV) dichloride (5\u2005mmol), dissolved in ethanol (30\u2005ml), was added to the solution which was further stirred for 2\u2005h. The precipitate that formed was filtered and then washed three times with cold ethanol to remove any impurities. The precipitate was then dried in a dessicator. The compound was crystallized in a mixture of chloro\u00adform and ethanol (1:2 v/v) at room temperature to give colourless slabs. Yield: 66%, m.p. 333\u2013336\u2005K. Analysis. Found C, 40.3; H, 7.3; N, 5.0; S, 22.8. C18H38N2O2S4Sn requires: C, 38.5; H, 6.8; N, 5.0; S, 23.7. IR (cm\u22121): 1490 \u03bd(C\u2014N), 991 \u03bd(C\u2014S), 553 \u03bd(Sn\u2014C), 420 \u03bd(Sn\u2014S). 1H NMR (CDCl3): 7.40\u20137.74 , 4.07 , 3.71 , 3.46 , 3.40 , 2.04 , 1.92 , 1.44 , 0.98 . 13C{1H} NMR (CDCl3): \u03b4 201.2 (S2C), 70.1 (OCH2), 59.1 (NCH2), 56.6 (OCH3), 44.5 (NCH3), 34.3 (SnCH2), 28.6 (SnCH2CH2), 26.5 (CH2CH3), 13.9 (CH2CH3). 119Sn{1H} NMR (CDCl3): 338.6.(2-Meth\u00adoxy\u00adeth\u00adyl)methyl\u00adamine (10\u2005mmol) dissolved in ethanol (30\u2005ml) was stirred in an ice bath (Uiso(H) set to 1.2\u20131.5Ueq(C). The mol\u00adecule has crystallographic mirror symmetry with the Sn atom and n-butyl-C atoms lying on the plane. The terminal CH2CH3 residue of each n-butyl group is statistically disordered across this plane. Owing to poor agreement, three reflections, i.e. (172), (124) and (155), were omitted from the final cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989017001098/hb7654sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017001098/hb7654Isup2.hklStructure factors: contains datablock(s) I. DOI: 1528948CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the complex mol\u00adecules inter\u00adact with the caffeine mol\u00adecules through O\u2014H\u22efN, O\u2014H\u22efO, C\u2013H\u22efS hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions.In the structure of the title compound, , [Co(NCS)2(H2O)4]\u00b72C8H10N4O2\u00b74H2O, the cobalt(II) cation lies on an inversion centre and is coordinated in a slightly distorted octa\u00adhedral geometry by the oxygen atoms of four water mol\u00adecules and two N atoms of two trans-arranged thio\u00adcyanate anions. In the crystal, the complex mol\u00adecules inter\u00adact with the caffeine mol\u00adecules through O\u2014H\u22efN, O\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid-to-centroid distance = 3.4715\u2005(5)\u2005\u00c5], forming layers parallel to the ab plane, which are further connected into a three-dimensional network by O\u2014H\u22efO and O\u2014H\u22efS hydrogen bonds involving the non-coordinating water mol\u00adecules.In the structure of the title compound [systematic name: tetra\u00adaqua\u00adbis\u00ad(thio\u00adcyanato-\u03ba The cobalt(II) cation lies on an inversion centre and displays a trans-arranged octa\u00adhedral coordination geometry provided by the N atoms of two thio\u00adcyanate anions and four O atoms of coordinating water mol\u00adecules. The Co1\u2014N15 [2.0981\u2005(8)\u2005\u00c5] and Co1\u2014O18 [2.0981\u2005(7)\u2005\u00c5] bond lengths are equal within standard uncertainties and significantly longer than the Co1\u2013O19 bond length [2.0732\u2005(7)\u2005\u00c5], and therefore the CoN2O4 octa\u00adhedron is slightly axially compressed. This structural feature is typical for related compounds \u2005\u00c5; Cg is the centroid of the N3/N7/C4/C6/C8/C9 ring; symmetry code: (i) 1\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z; Fig.\u00a02In the crystal, each complex mol\u00adecule inter\u00adacts with four neighboring caffeine mol\u00adecules through classical O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds Table\u00a01 involvin2\u00b76H2O in 5\u2005ml of water and caffeine in 10\u2005ml of ethanol was added to a solution of potassium thio\u00adcyanate in 5\u2005ml of water. Single crystals of the title compound suitable for X-ray analysis were grown after several months by slow evaporation of the solvent at room temperature.In a glass tube, a solution of CoClUiso(H) set at 1.2\u20131.5 times of the Ueq of the parent atom, after which the positions were refined with riding constraints global, I. DOI: 10.1107/S2056989017008180/rz5216Isup2.hklStructure factors: contains datablock(s) I. DOI: 1553654CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In this study, we investigated the proliferation and activation of lung \u03b3\u03b4 T cell subsets, specifically the IL-17 and IFN\u03b3 production by them following Cm lung infection. Our results found that five \u03b3\u03b4 T cell subsets, V\u03b31+ T, V\u03b32+ T, V\u03b34+ T, V\u03b35+ T, and V\u03b36+ T, expressed in lungs of na\u00efve mice, while Cm lung infection mainly induced the proliferation and activation of V\u03b34+ T cells at day 3 p.i., following V\u03b31+ T cells at day 7 p.i. Cytokine detection showed that Cm lung infection induced IFN\u03b3 secretion firstly by V\u03b34+ T cells at very early stage (day 3) and changed to V\u03b31+ T cells at midstage (day 7). Furthermore, V\u03b34+ T cell is the main \u03b3\u03b4 T cell subset that secretes IL-17 at the very early stage of Cm lung infection and V\u03b31+ T cell did not secrete IL-17 during the infection. These findings provide in vivo evidence that V\u03b34+T cells are the major IL-17 and IFN\u03b3-producing \u03b3\u03b4 T cell subsets at the early period of Cm lung infection.Our previous studies showed that C. pneumoniae causes respiratory diseases like bronchitis, sinusitis, and pneumonia, whereas C. trachomatis is a major cause of ocular and sexually transmitted diseases and 2\u2009mg/ml collagenase type XI ) for 60\u2009min at 37\u00b0C and added 2\u2009mM EDTA 5\u2009min before incubation finished. Then the cell population was purified by mixing with 35% Percoll (Sigma-Aldrich) and centrifuged for 20\u2009min at 750\u2009g, followed by lysis of erythrocytes with ammonium chloride-potassium (ACK) lysis buffer . The cells were washed twice using RPMI 1640 with 2% fetal calf serum and resuspended in complete RPMI 1640 medium (containing 10% FBS) for further experiment.Lung mononuclear cells were prepared as described previously . Briefly\u03b3 transcripts, total RNA was extracted from frozen lung tissues using Trizol agent (Invitrogen) according to the manufacturer's instruction. The isolated total RNA was reversely transcribed into cDNA (TaKaRa). Special primers for V\u03b31, V\u03b32, V\u03b34, V\u03b35, V\u03b36, and V\u03b37 were used to amplify cDNA. And \u03b2-actin, a housekeeping gene, was used as a control. The primers used in the PCR analysis were as follows: V\u03b31 (320\u2009bp), forward: 5\u2032-ACACAGCTATACATTGGTAC-3\u2032, reverse: 5\u2032-CTTATGGAGATTTGTTTCAGC-3\u2032; V\u03b32 (270\u2009bp), forward: 5\u2032-CGGCAAAAAACAAATCAACAG-3\u2032, reverse: 5\u2032-CTTATGGAGATTTGTTTCAGC-3\u2032; V\u03b34 (310\u2009bp), forward: 5\u2032-TGTCCTTGCAACCCCTACCC-3\u2032, reverse: 5\u2032-CTTATGGAGATTTGTTTCAGC-3\u2032; V\u03b35 (300\u2009bp), forward: 5\u2032-TGTGCACTGGTACCAACTGA-3\u2032, reverse: 5\u2032-CTTATGGAGATTTGTTTCAGC-3\u2032; V\u03b36 (300\u2009bp), forward: 5\u2032-TGTGCACTGGTACCAACTGA-3\u2032, reverse: 5\u2032-CTTATGGAGATTTGTTTCAGC-3\u2032; V\u03b37 (380\u2009bp), forward: 5\u2032-AAGCTAGAGGGGTCCTCTGC-3\u2032, reverse: 5\u2032-CTTATGGAGATTTGTTTCAGC-3\u2032; \u03b2-actin (582\u2009bp), forward: 5\u2032-CTTATGGAGATTTGTTTCAGC-3\u2032, reverse: 5\u2032-ATGAGGTAGTCMGTCAGGT-3\u2032. The products were electrophoresed in 1% agarose gel containing Gel-Red (0.01%). The bands were visualized and photographed by automatic gel imaging system and were analyzed for density on Image J software.To analyze the expressions of TCR V\u03b3\u03b4, anti-CD69, anti-TCRV\u03b31, anti-TCRV\u03b34, and isotype control Abs for 30\u2009min on ice for surface marker analysis. For intracellular cytokine analysis, single cell suspensions were stimulated with PMA (50\u2009ng/ml)/ionomycin (1\u2009\u03bcg/ml) (Sigma) for 6 hours at 37\u00b0C in the presence of 20\u2009mg/ml brefeldin A (Sigma). After the stimulation, cells were washed with FACS buffer twice and incubated with Fc receptor (FcR) block antibodies for 15\u2009min on ice to block nonspecific staining. Surface markers were stained first. The cells were then fixed with 4% w/v paraformaldehyde in PBS and permeabilized with permeabilization buffer , subsequently stained with anti-IFN\u03b3, IL-17, or corresponding isotype control Abs (eBioscience). The raw data were collected using FACS CantoII flow cytometer (BD Biosciences) and were analyzed using Flowjo 6.0 software.Lung mononuclear cells were aseptically prepared from mice at different time points postinfection and incubated with anti-CD3, anti-TCRt-test were used to determine statistical significance among groups. IFUs of Cm were converted to logarithmic values and analyzed using ANOVA. The value of p < 0.05 was considered as a statistically significant difference.One-way analysis of variance (ANOVA) and unpaired \u03b3\u03b4 T cells are the vital components of the innate immune system and play important roles in the early responses to pathogens. Our previous studies have shown that \u03b3\u03b4 T cells are the major producer of IL-17A in the very early stages of infection and depletion of \u03b3\u03b4 T cells by administration of mAb (GL3) against TCR\u03b3\u03b4 i.n. exists more body weight loss following Cm lung infection. The results here keep consistent with our previous studies that the percentage and absolute number of lung \u03b3\u03b4 T cells significantly increased at day 3 postinfection (p.i.) and reached to the highest level at day 7 p.i. Even though the percentage of \u03b3\u03b4 T cells reduced to baseline levels, the absolute number of \u03b3\u03b4 T cells still kept in a relatively higher level . All these results implicated that \u03b3\u03b4 T cells contribute to the IFN\u03b3 and IL-17 production and reduce morbidity during Cm infection, but its role in bacterial clearance is rather limited. Figures . CD69 wa Figures . TCR\u03b4\u2212/\u2212o 6 p.i. ; howeverfference . Further\u03b3\u03b4 T cells are heterogeneous population that can be subdivided based on the expression of specific V\u03b3 and V\u03b4 TCR chains. Although we already demonstrated the importance of \u03b3\u03b4 T cell in the early protection against Cm lung infection, this did not prove that \u03b3\u03b4 T cell subpopulation actually contributes to the \u03b3\u03b4 T cell-mediated early protection. To investigate this, we first analyzed the \u03b3\u03b4 T cell subsets in lungs of naive mice. Our results by using RT-PCR detection showed that there are more than five subpopulations, V\u03b31+ T, V\u03b32+ T, V\u03b34+ T, V\u03b35+ T, and V\u03b36+ T but not V\u03b37+ T cells; in lungs of naive mice, the expression intensity of mRNA is V\u03b32\u2009>\u2009V\u03b34\u2009>\u2009V\u03b31\u2009>\u2009V\u03b36\u2009>\u2009V\u03b35 while V\u03b31 T cells at midstage p.i. (day 7) against TCR\u03b3\u03b4 i.n. exists more body weight loss following Cm lung infection, which suggested that \u03b3\u03b4 T cells played a protective role in mice Chlamydia lung infection [\u03b4\u2212/\u2212 mice in this paper. It is worth mentioning that \u03b3\u03b4 T cell is the highest producer of IL-17A but the protection conferred by IL-17A is mainly mediated by Th17 cells following Cm infection. Therefore, the protective role of early production of IL-17 and IFN\u03b3 by V\u03b34+ T and V\u03b31+ T cells is not essential but supplementary in clearance of Chlamydia. In our present study using Cm infection model, it was found that V\u03b34+ T cells were the major source of IL-17 in the early stage, and V\u03b31+ T cells did not secrete IL-17. Similarly, Listeria monocytogenes also induces \u03b3\u03b4 T cells, especially V\u03b34+ T and V\u03b36+ T cells, and secretes IL-17 in infected liver, but more than 60 percent of the IL-17 are produced by V\u03b36+ T cell, which have fast kinetic response characteristics [\u03b31+ \u03b3\u03b4 T-cell reactivity which dominantly produces IL-17. Furthermore, with anti-CD3 antibody and virus-LPS stimulation in vitro, V\u03b31+ T cells dramatically produced IL-17, while only IL-10+ V\u03b34+ T cells existed [\u03b3\u03b4 T cell in varieties of pathogen infections are not always the same pattern, while these data suggest increased numbers of \u03b3\u03b4 T cells with cytokine-producing potential during immune response; any role for \u03b3\u03b4 T cell-derived cytokines in various model remains to be defined.The effector functions of le genes . IL-17-pfections , 36, 37.nfection . These reristics . However existed . Unlike \u03b34-\u03b3\u03b4 T cell subsets which is not identified during Cm infection. Interestingly, in our model, V\u03b36+ cells also present to proliferate following the Cm infection at the middle stage, which might be an important IL-17-producing cell after the early infection stage. IL-17+ V\u03b36+ T cells promote cancer cell growth by mobilizing peritoneal macrophages in the mice model of ovarian cancer [Listeria monocytogenes, more than 60 percent of the IL-17 are produced by V\u03b36+ T cell in infected liver, which have fast kinetic response characteristics [\u03b36+ T cell because it is reported that V\u03b36+ cells are the major \u03b3\u03b4 T cell population in reproductive tract but not in lungs [\u03b3 and IL-17 may be partially secreted by V\u03b36+ T cells apart from V\u03b31+ T and V\u03b34+ T cells during Cm infection.Notably, there are still a small number of IL-17-producing-Vn cancer . In Listeristics . In thisin lungs . But it \u03b31+ T and V\u03b34+ T cells are the major proliferative cell subsets of \u03b3\u03b4 T cell during Cm lung infection in mice. Moreover, V\u03b34+ T cells are the major IL-17 and IFN\u03b3-producing \u03b3\u03b4 T cell subsets at the early period of Cm lung infection. The findings in the present study provide new insights into the mechanisms bridging innate and adaptive immunity during lung chlamydial infections, which may have implications in developing effective chlamydial vaccines and in the understanding of host defense mechanisms in other lung infections.In conclusion, our data show that V"} +{"text": "C(4) along [100] and and There are two mol\u00adecules in the asymmetric unit of the title compound, one of them being disordered over the methyl group. The mol\u00adecules are linked by weak H\u22efS inter\u00adactions into chains with graph-set motifs 13H17N3S, one of them being disordered over the methyl group [site-occupancy ratio = 0.705\u2005(5):0.295\u2005(5)]. The maximum r.m.s. deviations from the mean plane of the non-H atoms for the tetra\u00adlone fragments amount to 0.4572\u2005(17) and 0.4558\u2005(15)\u2005\u00c5. The N\u2014N\u2014C\u2014N fragments are not planar and torsion angles are \u22129.4\u2005(2) and 8.3\u2005(2)\u00b0. In the crystal, the mol\u00adecules are linked by weak N\u2014H\u22efS inter\u00adactions into chains along [100] with graph-set motif C(4) and connected by weak N\u2014H\u22efS and C\u2014H\u22efS inter\u00adactions, forming R21(10) rings. The Hirshfeld surface analysis indicates that the most important contributions for the crystal packing are the H\u22efH (64.20%), H\u22efS (12.60%) and H\u22efC (12.00%) inter\u00adactions. The crystal packing resembles a herringbone arrangement when viewed along [001].There are two crystallographically independent mol\u00adecules in the asymmetric unit of the title compound, C In the 1940s it was reported that in in vitro assays, the thio\u00adsemicarbazone turned out to be very effective against tuberculosis. In contrast, the related oxygen-containing semicarbazones did not shown biological activity in the same assays, so that the sulfur atom in the mol\u00adecular structure is essential for Mycobacterium tuberculosis growth inhibition, a true milestone in the thio\u00adsemi\u00adcarbazone pharmacological research :0.295\u2005(5) with ed Fig.\u00a01.For the first mol\u00adecule, the C1/C2/C5/C10 atoms are essentially planar and atoms C3 and C4 deviate by 0.564\u2005(2) and \u22120.142\u2005(2)\u2005\u00c5, respectively, from this plane. For the second, the C14/C15/C18/C23 atoms are essentially planar while atoms C16 and C17 deviate from the plane by \u22120.534\u2005(2) and 0.201\u2005(2)\u2005\u00c5, respectively.In addition, the N1\u2014N2\u2014C11\u2014N3 and N4\u2014N5\u2014C24\u2014N6 torsion angles are 9.4\u2005(2)\u00b0 and 8.3\u2005(2)\u00b0. The dihedral angle between the tetra\u00adlone fragments of the two mol\u00adecules within the asymmetric unit is 85.51\u2005(02)\u00b0.i inter\u00adactions into chains along [100]. The S1\u2013C11\u2013N3\u2013H12 and S2\u2013C24\u2013N6\u2013H29 fragments are the subunits of the periodic arrangement, with graph-set motif C(4). In addition, the mol\u00adecules are linked by C9\u2014H10\u22efS2 and C22\u2014H27\u22efS1i inter\u00adactions building rings with graph-set motif D\u2014H\u22efS inter\u00adactions and di (x axis) values are the closest external and inter\u00adnal distances (in \u00c5) from given points on the Hirshfeld surface contacts hydrazinecarbo\u00adthio\u00adamide, the mol\u00ad\u00adecules are linked into chains by N\u2014H\u22efS hydrogen bonds (H\u22efS distances = 2.45 and 2.71\u2005\u00c5) and the H\u22efS contribution for the cohesion of the structure amounts to 19.20% . This kind of arrangement, the one-dimensional hydrogen-bonded polymer, is possible due to the unsubstituted amine, which increases the possibilities for inter\u00admolecular hydrogen bonding hydrazinecarbo\u00adthio\u00adamide, one H atom of the amine group is substituted by one methyl group. The N\u2014H\u22efS hydrogen bonds are weaker in comparison with the first structure (H\u22efS distances = 3.03 and 3.29\u2005\u00c5), the H\u22efS contribution for the cohesion of the structure amounts to 15.80% and the dimensionality of the structure is preserved with mol\u00adecules linked into chains . The disorder over the mol\u00adecules in the asymmetric unit was not considered and the calculations were performed using atoms of the major occupancy component hydrazinecarbo\u00adthio\u00adamide, the mol\u00adecules are also linked by N\u2014H\u22efS hydrogen bonds, but not into hydrogen-bonded polymers (H\u22efS distance = 2.70\u2005\u00c5). The phenyl rings linked to the amino groups change the mol\u00adecular arrangement due to steric effects: the mol\u00adecules build dimers and the H\u22efS contribution to the crystal packing amounts to 13.00% . Thus, there is a relationship between the mol\u00adecular assembly, the geometry of the H\u22efS inter\u00adactions and their contribution to the crystal structures and two positions will be possible for the terminal CH3\u2013group. The C25 and C26 atoms were fixed with restraints (SADI command) and had to be split over two positions. The occupancy factor for C25A and C26A is 0.705\u2005(5) and for C25B and C26B it is 0.295\u2005(5). H atoms were located in difference maps but were positioned with idealized geometry and were refined with isotropic displacement parameters using a riding model (HFIX command) with Uiso(H) = 1.2Ueq(secondary C atoms) , Uiso(H) = 1.5 Ueq (C\u2014H = 0.98\u2005\u00c5) and Uiso(H) = 1.2 Ueq(N) (N\u2014H = 0.88\u2005\u00c5). Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017001311/lh5835sup1.cifCrystal structure: contains datablock(s) I, publication_text. DOI: 10.1107/S2056989017001311/lh5835Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017001311/lh5835Isup3.cmlSupporting information file. DOI: 1529622CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "By contrast, analogous syntheses from o\u2010xylene and m\u2010xylene as the solvent yield the solvent\u2010free coordination polymer [Ag4(O2CCF3)4(phen)2] (2). Toluene, p\u2010xylene and benzene have been successfully used in mixed\u2010arene syntheses to template the formation of coordination polymers 1\u22c5phen\u22c5arene, which incorporate o\u2010 or m\u2010xylene. The selectivity of 1\u22c5phen\u22c5arene for the arene guests was determined, through pairwise competition experiments, to be p\u2010xylene>toluene\u2248benzene>o\u2010xylene>m\u2010xylene. The largest selectivity coefficient was determined as 14.2 for p\u2010xylene:m\u2010xylene and the smallest was 1.0 for toluene:benzene.The coordination polymers [Ag Materials that are porous on the molecular scale have been in use for many years in applications involving molecular separation. Fixed\u2010pore materials, exemplified by zeolites and related inorganic porous materials,4{O2C(CF2)2CF3}4(phen)2(arene)n]\u22c5m\u2009(arene) (phen=phenazine) and examined the role of these guests in templating solid\u2010state transformations.o\u2010xylene, m\u2010xylene and p\u2010xylene by the coordination polymer [Ag4(O2CCF3)4(phen)3] (1) during its self\u2010assembly from the solution phase. This results in the crystalline materials [Ag4(O2CCF3)4(phen)3]\u22c5phen\u22c5arene (1\u22c5phen\u22c5arene), in which the arenes act as co\u2010guests alongside non\u2010coordinated phenazine. Specifically, we are able to establish the selectivity of the coordination polymer for each of the five arenes by determination of the pairwise selectivity coefficients, and are able to determine the potential of this material in separation of structurally similar arenes by recycling of the encapsulation process.We have developed a class of 1D coordination polymers based on silver(I) perfluoroalkylcarboxylate dimer units linked through diimine ligands, such as substituted pyrazine or phenazine is important commercially due to the large scale on which such compounds are synthesised for use as precursors in the chemical industry, combined with the non\u2010regiospecific manner in which alkylarenes, such as xylenes, are synthesised. Their similarity in physical properties makes conventional methods of separation, such as distillation, a difficult and not very cost\u2010effective approach.p\u2010xylene or benzene resulted in exclusive formation of the corresponding arene\u2010guest\u2010containing 1D coordination polymer [Ag4(O2CCF3)4(phen)3]\u22c5phen\u22c5arene (1\u22c5phen\u22c5arene). Phase purity was confirmed by elemental analysis and Pawley fitting of the corresponding X\u2010ray powder patterns. Crystal structures of [Ag4(O2CCF3)4(phen)3]\u22c5phen\u22c52\u2009(toluene) (1\u22c5phen\u22c5tol), [Ag4(O2CCF3)4(phen)3]\u22c5phen\u22c52\u2009(p\u2010xylene) (1\u22c5phen\u22c5pxyl) and [Ag4(O2CCF3)4(phen)3]\u22c5phen\u22c52\u2009C6H6 (1\u22c5phen\u22c5benz) were determined by single\u2010crystal X\u2010ray diffraction 4(L)3] described in our previous work.1\u22c5phen\u22c5arene material additional non\u2010coordinated phenazine molecules are included as guests, situated between each of the doubly\u2010bridging phenazine linkers in a \u03c0\u2010stacked manner. Two equivalents of the arene used as solvent are also present as guests per repeat unit of the polymer. These molecules are \u03c0\u2010stacked on both sides of the electron\u2010deficient central ring of the singly\u2010bridging phenazine ligands. The arenes are crystallographically ordered and each arene molecule is related to another by a centre of symmetry located in the centre of those phenazine ligands trifluoroacetate in methanol onto a solution of phenazine dissolved in either toluene, o\u2010 and m\u2010xylene, however, did not yield the analogous 1D coordination polymer. Rather, these syntheses led exclusively to the 2D coordination polymer [Ag4(O2CCF3)4(phen)2] (2), which excludes the xylene guests. This more densely\u2010packed phase is propagated in one dimension by an extended arrangement of silver perfluoroacetate units that employ both the anti and syn lone pairs on the carboxylate oxygen in coordination to AgI centres,Analogous syntheses conducted using p\u2010xylene and benzene) were included in the self\u2010assembly of 1\u22c5phen\u22c5arene, the selectivity of this inclusion process was examined by means of pairwise competition experiments between the three arenes. This was achieved by conducting the assembly of 1\u22c5phen\u22c5arene in the presence of a 1:1 mixture (by volume) of two of the three possible pairs of arenes. Pawley fitting of X\u2010ray powder diffraction confirmed the formation of 1\u22c5phen\u22c5arene, along with a very small amount of 2.6]DMSO, and studying the resulting solution by 1H\u2005NMR spectroscopy and gas chromatography (GC). These data and their analyses are presented in full in the Supporting Information. Pairwise selectivity constants, KA:B were determined from the corresponding inclusion experiments \u22c5phen\u22c52\u2009(toluene) (1\u22c5phen\u22c5tol)\u22c5phen\u22c52\u2009(p\u2010xylene) (1\u22c5phen\u22c5pxyl)\u22c5phen\u22c52\u2009C6H6 (1\u22c5phen\u22c5benz)\u22c5phen\u22c52\u2009{(toluene)0.73\u22c5(p\u2010xylene)0.27} (1\u22c5phen\u22c5 tol\u22c5pxyl)\u22c5phen\u22c52\u2009{(toluene)0.49\u22c5(p\u2010xylene)0.51} (1\u22c5phen\u22c5 tol\u22c5pxyl)\u22c5phen\u22c52\u2009{(toluene)0.38\u22c5(p\u2010xylene)0.62} (1\u22c5phen\u22c5 tol\u22c5pxyl)\u22c5phen\u22c52\u2009{(toluene)0.30\u22c5(p\u2010xylene)0.70} (1\u22c5phen\u22c5 tol\u22c5pxyl)\u22c5phen\u22c52\u2009{(toluene)0.14\u22c5(p\u2010xylene)0.86} (1\u22c5phen\u22c5 tol\u22c5pxyl)\u22c5phen\u22c52\u2009{(toluene)0.46\u22c5(C6H6)0.54} (1\u22c5phen\u22c5tol\u22c5 benz)\u22c5phen\u22c52\u2009{(p\u2010xylene)0.58\u22c5(C6H6)0.42} (1\u22c5phen\u22c5 pxyl\u22c5benz)\u22c5phen\u22c52\u2009{(C6H6)0.81\u22c5(o\u2010xylene)0.19} (1\u22c5phen\u22c5 C6H6\u22c5oxyl)[Ag: A 0.05\u2009m solution of AgO2CCF3 in methanol (8\u2005mL) was layered on to a 0.05\u2009m solution of phenazine in 1:1 (v/v) benzene:o\u2010xylene (8\u2005mL). Yield 40\u2009% .4(O2CCF3)4(phen)3]\u22c5phen\u22c52\u2009{(C6H6)0.91\u22c5(m\u2010xylene)0.09} (1\u22c5phen\u22c5C6H6\u22c5mxyl)[Ag: A 0.05\u2009m solution of AgO2CCF3 in methanol (8\u2005mL) was layered on to a 0.05\u2009m solution of phenazine in 1:1 (v/v) benzene:m\u2010xylene (8\u2005mL) . Yield 40\u2009% .4(O2CCF3)4(phen)3]\u22c5phen\u22c52\u2009{(toluene)0.84\u22c5(o\u2010xylene)0.16} (1\u22c5phen\u22c5 tol\u22c5oxyl)[Ag: A 0.05\u2009m solution of AgO2CCF3 in methanol (8\u2005mL) was layered on to a 0.05\u2009m solution of phenazine in 1:1 (v/v) toluene:o\u2010xylene (8\u2005mL). Yield 33\u2009% .4(O2CCF3)4(phen)3]\u22c5phen\u22c52\u2009{(toluene)0.91\u22c5(m\u2010xylene)0.09} (1\u22c5phen\u22c5tol\u22c5mxyl)[Ag: A 0.05\u2009m solution of AgO2CCF3 in methanol (8\u2005mL) was layered on to a 0.05\u2009m solution of phenazine in 1:1 (v/v) toluene:m\u2010xylene (8\u2005mL). Yield 32\u2009% .4(O2CCF3)4(phen)3]\u22c5phen\u22c52\u2009{(p\u2010xylene)0.90\u22c5(o\u2010xylene)0.10} (1\u22c5phen\u22c5pxyl\u22c5oxyl)[Ag: A 0.05\u2009m solution of AgO2CCF3 in methanol (8\u2005mL) was layered on to a 0.05\u2009m solution of phenazine in 1:1 (v/v) p\u2010xylene:o\u2010xylene (8\u2005mL). Yield 42\u2009% .4(O2CCF3)4(phen)3]\u22c5phen\u22c52\u2009{(p\u2010xylene)0.96\u22c5(m\u2010xylene)0.04} (1\u22c5phen\u22c5pxyl\u22c5mxyl)[Ag: A 0.05\u2009m solution of AgO2CCF3 in methanol (8\u2005mL) was layered on to a 0.05\u2009m solution of phenazine in 1:1 (v/v) p\u2010xylene:m\u2010xylene (8\u2005mL). Yield 45\u2009% .2(O2CCF3)2(phen)] (2)DMSO, then filtered through cotton wool. 1H\u2005NMR spectra were measured on a Bruker AV 400\u2005MHz spectrometer. The NMR spectra can be found in the Supporting Information, Section 4. The NMR spectra were analysed using the Bruker TOPSPIN 3.1 programme. Methyl peaks for mixed xylene systems, which did not show complete baseline separation, were deconvoluted using the mixed\u2010line descriptor (mixed Lawrencian & Gaussian) deconvolution function in TOPSPIN.Gas chromatography: The solutions used for 1H\u2005NMR were transferred to glass vials using crimped caps, and then analysed using a PerkinElmer Autosystem GC with an AlltechTM HeliflexTM AT\u20101 capillary column (L\u00d7I.D. 30\u2005m\u00d70.32\u2005mm\u00d7df 5.00\u2005\u03bcm), heating from 40 to 200\u2009\u00b0C at 10\u2009\u00b0C min\u22121. Expected guest retention times were found to be 9.9\u2005min (benzene), 12.7\u2005min (toluene), 15.1\u2005min (p\u2010xylene), 15.2\u2005min (m\u2010xylene\u2010indistinguishable from p\u2010xylene) and 15.7\u2005min (o\u2010xylene). Relative content of guests was determined by direct comparison of chromatogram peak areas. The gas chromatograms can be found in the Supporting Information.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "In their respective crystals, hydrogen bonding leads to dimeric aggregates in the former (O\u2014H\u22efS) and supra\u00admolecular chains in the latter .Highly-distorted five-coordinate NS 5H10NS2)2(C5H5NO)], (I), and [Zn(C4H8NOS2)2(C5H5NO)], (II), are NS4 donor sets derived from N-bound hy\u00addroxy\u00adpyridyl ligands and asymmetrically chelating di\u00adthio\u00adcarbamate ligands. The resulting coordination geometries are highly distorted, being inter\u00admediate between square pyramidal and trigonal bipyramidal for both independent mol\u00adecules comprising the asymmetric unit of (I), and significantly closer towards square pyramidal in (II). The key feature of the mol\u00adecular packing in (I) is the formation of centrosymmetric, dimeric aggregates sustained by pairs of hy\u00addroxy-O\u2014H\u22efS(di\u00adthio\u00adcarbamate) hydrogen bonds. The aggregates are connected into a three-dimensional architecture by methyl\u00adene-C\u2014H\u22efO(hy\u00addroxy) and methyl-C\u2014H\u22ef\u03c0(chelate) inter\u00adactions. With greater hydrogen-bonding potential, supra\u00admolecular chains along the c axis are formed in the crystal of (II), sustained by hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) hydrogen bonds, with ethyl\u00adhydroxy and pyridyl\u00adhydroxy groups as the donors, along with ethyl\u00adhydroxy-O\u2014H\u22efS(di\u00adthio\u00adcarbamate) hydrogen bonds. Chains are connected into layers in the ac plane by methyl\u00adene-C\u2014H\u22ef\u03c0(chelate) inter\u00adactions and these stack along the b axis, with no directional inter\u00adactions between them. An analysis of the Hirshfeld surfaces clearly distinguished the independent mol\u00adecules of (I) and reveals the importance of the C\u2014H\u22ef\u03c0(chelate) inter\u00adactions in the packing of both (I) and (II).The common feature of the mol\u00adecular structures of the title compounds, [Zn(C Zn(S2CNEt2)2]2(bpe)\u00b7bpe 2]2(bpe)}n, for R = Et and n-Bu 2]2(bpe) S2CN(+)RR\u2032, with two formally negatively charged sulfur atoms, which makes di\u00adthio\u00adcarbamate a very effective chelating agent, thereby decreasing the Lewis acidity of the zinc atom.The structures of binary zinc bis\u00ad(di\u00adthio\u00adcarbamates) are always zero-dimensional (i.e using di\u00adthio\u00adcarbamate anions of the type \u2212S2CN(R)CH2CH2OH. This influence is seen in the recent report of the structures of Zn[S2CN(R)CH2CH2OH]2 for R = i-Pr and CH2CH2OH 2 CH2CH2OH]2, in the case when R = CH2CH2OH, isolated as a 1:1 hydrate, leads to supra\u00admolecular ladders and these extend in two dimensions via water-O\u2014H\u22efS(di\u00adthio\u00adcarbamate) hydrogen bonds. When R = i-Pr, layers are sustained by hy\u00addroxy-O\u2014H\u22efS hydrogen bonds 2 has been complexed with 3-hy\u00addroxy\u00adpyridine (pyOH) to yield two 1:1 complexes. Quite different aggregation patterns are observed when R = R\u2032 = Et (I)R = i-Pr and R\u2032 = CH2CH2OH (II)An approach to increase the supra\u00admolecular aggregation in the crystal structures of zinc di\u00adthio\u00adcarbamates has been to introduce hydrogen bonding functionality into the ligands, 2CNEt2)2(pyOH) comprise the asymmetric unit of (I)a, the ZnII atom is chelated by two di\u00adthio\u00adcarbamate ligands and one nitro\u00adgen atom derived from the monodentate pyOH ligand. The S1-di\u00adthio\u00adcarbamate ligand chelates the zinc atom forming quite different Zn\u2014S bond lengths compared with the S3-di\u00adthio\u00adcarbamate ligand. This is qu\u00adanti\u00adfied in the values of \u0394(Zn\u2014S), being the difference between the Zn\u2014Slong and Zn\u2014Sshort bond lengths, Table\u00a01i.e. 0.43 and 0.15\u2005\u00c5, respectively. The Zn1\u2014N3 bond length is significantly shorter than the Zn\u2014S bonds. The NS4 coordination geometry is highly distorted as seen in the value of \u03c4 of 0.48 and, especially, the Sl\u2014Zn\u2014Sl (l = long) bond angles, Table\u00a01b, is quite similar to that just described for the Zn1 atom. The values of \u0394(Zn\u2014S) of 0.21 and 0.25\u2005\u00c5 are inter\u00admediate to those for the Zn1-mol\u00adecule. Even so, the differences in the Zn\u2014S bond lengths in both mol\u00adecules are not that great with this observation reflected in the closeness of the C\u2014S bond lengths, Table\u00a01cf. the Zn1-mol\u00adecule.Two independent mol\u00adecules of Zn(S2CN(Me)CH2CH2OH]2(pyOH), is shown in Fig.\u00a02The mol\u00adecular structure of (II)Overlay diagrams of the three mol\u00adecules in (I)via a 14-membered {\u22efHOC2NZnS}2 synthon, Fig.\u00a04a and Table\u00a02x, y, z. The aggregates are further stabilized by pyOH-C\u2014H\u22ef\u03c0 inter\u00adactions where the \u03c0-system is a chelate ring. Such C\u2014H\u22ef\u03c0(chelate) inter\u00adactions are increasingly being recognized as being important in the supra\u00admolecular chemistry of metal 1,1-di\u00adthiol\u00adates but with methyl-H atoms as the donors, Fig.\u00a04b. The connections between layers along the c direction are of the type methyl\u00adene-C\u2014H\u22efO(hy\u00addroxy), Fig.\u00a04c.The key feature of the mol\u00adecular packing of (I)a, these hydrogen bonds lead to a centrosymmetric 22-membered {\u22efSZnSCNC2OH\u22efOH}2 synthon. On either side of these synthons, the pyOH hy\u00addroxy group hydrogen bonds to the O2-hy\u00addroxy atom and through symmetry, a centrosymmetric 24-membered {\u22efOC2NCSZnNC2OH}2 synthon is formed, highlighted as \u20182\u2019 in Fig.\u00a05a. Alternating synthons generate a supra\u00admolecular chain aligned along the c axis. Methyl\u00adene-C\u2014H\u22ef\u03c0(chelate) inter\u00adactions link mol\u00adecules into dimeric units, Fig.\u00a05b. The combination of the aforementioned inter\u00adactions lead to supra\u00admolecular layers that stack along the b axis with no directional inter\u00adactions between them, Fig.\u00a05c.The addition of greater hydrogen-bonding potential in (II)et al., 2016dnorm in the range \u22120.2 to + 1.3 au for the Zn1- and Zn2-containing mol\u00adecules of (I)O and -H2O, and di\u00adthio\u00adcarbamate-S2 and S8 atoms represent the donors and acceptors of the O\u2014H\u22efS hydrogen bonds; these are viewed as blue and red regions on the Hirshfeld surfaces mapped over electrostatic potential (mapped over the range \u22120.07 to +0.10 au), Fig.\u00a07b and 6c are due to comparatively weaker inter\u00admolecular C\u2014H\u22efO inter\u00adactions. The intra-dimer \u03c0\u2013\u03c0 stacking inter\u00adaction between the pyOH rings, Fig.\u00a04a, is evident through the appearance of faint-red spots near the arene-C13 and C26 atoms of the rings, Fig.\u00a06a and 6b, forming a close inter\u00adatomic C\u22efC contact, Table\u00a04a\u2013c, characterize the influence of the C\u2014H\u22ef\u03c0(chelate) inter\u00adactions; in Fig.\u00a07The Hirshfeld surface analysis for (I)dnorm, Fig.\u00a09a and 9b, and result in the blue and red regions corres\u00adponding to positive and negative potential on the Hirshfeld surface mapped over electrostatic potential (mapped over the range \u22120.12 to +0.18 au), Fig.\u00a09c. The faint-red spots near the S4, C8, C9 and H2B atoms in Fig.\u00a09a and 9b indicate their involvement in short inter\u00adatomic S\u22efS, C\u22efC and C\u22efH/H\u22efC contacts, Table\u00a04a illustrates the immediate environment about a reference mol\u00adecule within Hirshfeld surfaces mapped over electrostatic potential and highlights the O\u2014H\u22efO hydrogen bonds. The C\u2014H\u22ef\u03c0(chelate) and its reciprocal contact, i.e. \u03c0\u2014H\u22efC, and short inter\u00adatomic S\u22efS, C\u22efC and C\u22efH/H\u22efC contacts, with labels 3\u20136, are shown in Fig.\u00a010b.The presence of peripheral hy\u00addroxy groups participating in the O\u2014H\u22efO hydrogen bonds in the structure of (II)a. The respective plots delineated into H\u22efH, O\u22efH/H\u22efO, S\u22efH/H\u22efS, C\u22efH/H\u22efC, C\u22efC and S\u22efS contacts The overall two-dimensional fingerprint plot for individual Zn1- and Zn2-containing mol\u00adecules, overall (I)b, show different distributions of points in the individ\u00adual plots for Zn1- and Zn2-mol\u00adecules. This, as well as their different percentage contributions to the Hirshfeld surface, Table\u00a05de, di) distances shorter than their van der Waals separations, corresponding to short inter\u00adatomic H\u22efH contacts, Table\u00a04The fingerprint plots delineated into H\u22efH contacts for (I)c, also exhibit slightly different profiles for the independent mol\u00adecules. The respective peaks at de + di \u223c 2.7\u2005\u00c5 and \u223c 2.6\u2005\u00c5 correspond to donors (upper region) and the acceptors (lower region) for the Zn1-mol\u00adecule, whereas these appear as a pair of peaks at the same de + di \u223c 2.6\u2005\u00c5 distance for the Zn2-mol\u00adecule. This is likely due to the inter\u00adacting oxygen and hydrogen atoms for the Zn1-mol\u00adecule being at their van der Waals separation in the donor region, i.e. at 2.72\u2005\u00c5, while in the acceptor region the peak corresponds to a short inter\u00adatomic O\u22efH contact, Table\u00a04de + di \u223c 2.6\u2005\u00c5.The fingerprint plots delineated into O\u22efH/H\u22efO contacts, Fig.\u00a011de + di distances in the fingerprint plots delineated into S\u22efH/H\u22efS contacts, Fig.\u00a011d, for the Zn1- and Zn2-mol\u00adecules result from different hy\u00addroxy-O\u2014H\u22efS(di\u00adthio\u00adcarbamate) hydrogen bonds. The tips at de + di \u223c 2.4\u2005\u00c5 in the donor region of the plot for the Zn1-mol\u00adecule and in the acceptor region for the Zn2-mol\u00adecule are due to the formation of O\u2014H\u22efS hydrogen bonds between the hy\u00addroxy-O1 and di\u00adthio\u00adcarbamate-S8 atoms; the other hydrogen bond, involving the O2 and S2 atoms, gives rise to tips at de + di \u223c 2.3\u2005\u00c5 in the respective donor and acceptor regions of the plots, Fig.\u00a011d. The plot for the overall structure results from the superimposition of individual plots and shows the symmetric distribution of points as a pair of long spikes having tips at de + di \u223c 2.3\u2005\u00c5. The short inter\u00adatomic S\u22efH/H\u22efS contacts in the crystal of (I)de + di \u223c 3.0\u2005\u00c5 in the respective plots.The pair of spikes with their tips at different e, have the same shape with tips at de + di \u223c 2.7\u2005\u00c5 which are due to the short inter\u00adatomic C\u22efH/H\u22efC contacts, Table\u00a04i.e. 1.8%, but recognizable contribution to the Hirshfeld surface and appear as an arrow-like distribution of points around de = di = 1.8\u2005\u00c5 in Fig.\u00a011f. As indicated in Fig.\u00a011g, S\u22efS contacts do not figure prominently in the mol\u00adecular packing of (I)Almost the same percentage contribution from C\u22efH/H\u22efC contacts to the overall surface is made by the Zn1- and Zn2-mol\u00adecules, Table\u00a05b, a pair of very thin spikes having their tips at de + di \u223c 2.3\u2005\u00c5 indicate the presence of short inter\u00adatomic H\u22efH contacts between hy\u00addroxy-H1O and -H2O atoms, Table\u00a04O and H3O atoms, Table\u00a04cf. (I)de + di \u223c 1.8\u2005\u00c5, Fig.\u00a011c. The tips corresponding to the O1\u22efH6A contact, Table\u00a04The corresponding two-dimensional fingerprint plots for (II)i.e. 22.2 and 22.7%, respectively, reflect the O\u2014H\u22efS hydrogen bonds and additional S\u22efH contacts resulting in tips at de + di \u223c 2.9\u2005\u00c5 in Fig.\u00a011d and Table\u00a04de + di \u223c 2.6\u2005\u00c5 in the plot, Fig.\u00a011e, results from the C\u2014H\u22ef\u03c0(chelate) and short inter\u00adatomic C\u22efH/H\u22efC contacts, Table\u00a04f, as the pair of tips at de + di \u223c1.7\u2005\u00c5. Finally, a cone-shaped distribution of points with a 3.8% contribution to the surface from S\u22efS contacts having a vertex at de = di \u223c 1.7\u2005\u00c5 in the fingerprint plot, Fig.\u00a011g, results from short inter\u00adatomic contacts between S4 atoms, Table\u00a04The S\u22efH/H\u22efS contacts with the nearly same contribution to the surface of (II)Chemical context, the presence of hydroxy\u00adethyl groups in zinc di\u00adthio\u00adcarbamates leads to a higher degree of recognizable supra\u00admolecular aggregation owing to hydrogen bonding, usually of the type hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) but, sometimes also of the type hy\u00addroxy-O\u2014H\u22efS(di\u00adthio\u00adcarbamate) 2]2}2, the usual dimeric motif is evident but these self-assemble via strong hydrogen bonding into three-dimensional architectures in both of the polymorphs characterized thus far, with the difference between the structures being the topology of supra\u00admolecular layers, i.e. flattened CH2CH2OH]2}2, the reduced hydrogen bonding leads to supra\u00admolecular chains CH2CH2OH)2]2}2L, where L is (3-pyrid\u00adyl)CH2N(H)C(=O)C(=O)N(H)CH2(3-pyrid\u00adyl). However, hydrogen bonding of the type hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) links the mol\u00adecules into inter-woven double chains 2, N,N\u2032-bis\u00ad(pyridin-3-ylmeth\u00adyl)ethane\u00addithiodi\u00adamide : \u03bd(C=N) 1482 ; \u03bd(C\u2014S) 987 (s). 1H NMR : \u03b4 9.91 , 8.20\u20138.00 , 7.30\u20137.10 , 3.82 ; 1.22 .Synthesis of (I)2N(Me)CH2CH2OH]2 and 3-hy\u00addroxy pyridine were dissolved in a MeOH/EtOH (1:1 v/v) solution. Solvent diffusion of hexane into this solution led to the formation of colourless crystals. FT\u2013IR (cm\u22121): \u03bd(C=N) 1480 (s); \u03bd(C\u2014S) 1002 (s). 1H NMR : \u03b4 9.91 , 8.20\u20138.00 , 7.30\u20137.10 , 4.91 ; 3.90 ; 3.70 ; 3.41 .Synthesis of (II)Uiso(H) set to 1.2\u20131.5Ueq(C). The oxygen-bound H-atoms were located in difference Fourier maps but were refined with a distance restraint of O\u2014H = 0.84\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S205698901601728X/hb7628sup1.cifCrystal structure: contains datablock(s) . DOI: 10.1107/S205698901601728X/hb7628Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S205698901601728X/hb7628IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1511865, 1511864CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A similar reaction of SnCl4 with 2 equiv. of Li[o-(C2H10B10)C(NiPr)(NHiPr)] unexpectedly afforded the known solvated penta\u00adchlorido\u00adstannate(IV) salt [Li(THF)4][SnCl5(THF)] as the main reaction product. Small amounts of the new chlorido-tin(IV) bis\u00ad(carboranylamidinate) bischloridotin(IV), SnIVCl[o-(C2H10B10)C(NiPr)(NHiPr)][o-(C2H10B10)C(NiPr)2] (2), were isolated as a by-product. Single-crystal X-ray structure analysis revealed a \u03baC,\u03baN-chelating coordination of the carboranylamidinate ligands in both 1 and 2. The Sn atom in 1 adopts a pseudo-trigonal\u2013bipyramidal coordination under participation of a stereoactive lone pair. In 2, a trigonal\u2013bipyramidal coordination of Sn is completed by a chlorido ligand.Reaction of 2 equiv. of the lithium carboranylamidinate Li[ RC(NR\u2032)2]\u2212 are the nitro\u00adgen analogs of carboxyl\u00adate anions. These versatile N,N\u2032-chelating ligands form stable coordination compounds with nearly every metallic element in the Periodic Table. In view of this rich coordination chemistry, amidinate ligands are frequently regarded as \u2018steric cyclo\u00adpenta\u00addienyl equivalents\u2019. Metal complexes comprising amidinato ligands are readily available by insertion of the 1,3-diorganocarbodi\u00adimide, R\u2013-N=C=N\u2014R, into an M\u2014C bond of an organometallic precursor compound. Another common synthetic route involves salt metathetical reactions between lithium amidinates and metal halides 2]\u2212 is the fact that the substituents R and R\u2032 attached to the N\u2013C\u2013N unit can be varied in many ways. With R = ortho-C2H11B10 (\u2018ortho-carboran\u00adyl\u2019) we introduced a sterically demanding and chemically versatile moiety in the backbone of the amidinate ligand. Carboranes are of tremendous scientific and technological inter\u00adest due to their various applications ever since their discovery in the 1960\u2032s. These applications include the synthesis of polymers and ceramics, catalysts, radiopharmaceuticals and non-linear optics, as well as the BNCT (= boron neutron capture therapy) technique C(NiPr)(NHiPr)]\u2212 (= [HiLPr]\u2212) was synthesized in our lab in 2010 by in situ li\u00adthia\u00adtion of the parent o-carborane, ortho-C2H12B10 (= ortho-dicarba-closo-dodeca\u00adborane), followed by treatment with 1 equiv. of N,N\u2032-diiso\u00adpropyl\u00adcarbodi\u00adimide , a proton is formally shifted from the carborane C atom to the amidinate unit, resulting in an amidine moiety that usually acts as a monodentate donor functionality as shown in Fig.\u00a02a.A key advantage of the amidinate anions [RL]2\u2013, whose favored coordination mode is still \u03baC,\u03baN . Derivatives of Si, P, Ge, SnII, SnIV, FeII and FeIII, RhI and IrI containing this ligand system have been prepared by double li\u00adthia\u00adtion of the parent ortho-carboranyl\u00adamidine followed by treatment with appropriate element chloride precursors C(NCy)(NHCy)]2 and SnIVCl[o-(C2H10B10)C(NiPr)(NHiPr)][o-(C2H10B10)C(NiPr)2] . Compound 2 is the first carboranylamidinate complex containing both mono- and dianionic carboranylamidinate ligands in a single mol\u00adecule.Among the known carboranylamidinate complexes are only very few compounds with more than one carboranylamidinate ligand per metal atom, and these are exclusively of the type ortho-carboranylamidinate precursors Li[HLCy] and Li[HiLPr] were readily available following a published procedure by reaction of the mono-li\u00adthia\u00adted o-carborane Li-o-C2B10H11 with a stoichiometric amount of the carbodi\u00adimides iPrN=C=NiPr or CyN=C=NCy, respectively, in THF as colorless, block-like single crystals after recrystallization from toluene. The low isolated yield of ca 20% can be traced back to the very high solubility of 1 even in non-polar organic solvents. In addition to the X-ray diffraction study, compound 1 was also characterized through elemental analysis and the usual set of spectroscopic methods. In the IR spectrum, a characteristic \u03bd(NH) band at 3423\u2005cm\u22121 confirmed the presence of monoanionic [HLCy]\u2212 ligands. The NH functionalities were also observed in the 1H NMR spectrum through a broad singlet at \u03b4 4.50 ppm. A single 119Sn NMR resonance at \u03b4 \u221246 ppm was in agreement with the formation of a single Sn-containing species. The mass spectrum of 1 showed the mol\u00adecular ion at m/z 818 with 27% relative intensity.The mono-li\u00adthio-4 with 2 equiv. of Li[HiLPr] was carried out with the aim of synthesizing the hitherto unknown tin(IV) bis\u00ad(carboranylamidinate) Sn[iLPr]2. Cooling of the reaction mixture afforded a fairly large amount of well-formed, colorless crystals, which turned out to be the known solvated penta\u00adchlorido\u00adstannate(IV) salt [Li(THF)4][SnCl5(THF)]. This compound was first prepared and structurally characterized by Junk & Leary (2000ca 5% isolated yield) of the unexpected tin(IV) carboranylamidinate SnCl[iLPr][HiLPr] (2) could be obtained. The X-ray crystal structure determination of 2 revealed the presence of the first complex containing both a mono- and a dianionic carboranylamidinate ligand in one mol\u00adecule. As in 1, the IR spectrum of 2 showed a characteristic \u03bd(NH) band at 3410\u2005cm\u22121. Elemental analysis and a single resonance in the 119Sn NMR spectrum (\u03b4 290 ppm) confirmed the purity of 2. In the mass spectrum, the mol\u00adecular ion was observed at m/z 692 with 47% relative intensity.A similar reaction of SnCleary 2000. From th1 and 2 are illustrated in Figs.\u20053\u03baC,\u03baN-chelating mode. In the tin(II) complex Sn[HLCy]2 (1), the coordination of the metal atom can be described as pseudo-trigonal\u2013bipyramidal, with the two N atoms defining the axial positions [N1\u2014Sn\u2014N3 157.76\u2005(4)\u00b0]. The two Sn-C bonds are situated in the equatorial plane [C3\u2014Sn\u2014C18 100.12\u2005(5)\u00b0], together with the stereoactive lone pair at Sn. The bite angles of the [HLCy]\u2212 ligands are 72.01\u2005(5)\u00b0 (N1\u2014Sn\u2014C3) and 73.27\u2005(5)\u00b0 (N3\u2014Sn\u2014C18). The coordination geometry of the Sn atom therefore resembles that in the related tin(II) bis\u00ad(carboranylamidinate) Sn[HLiPr]2 . These values are in the same range as those observed in the isopropyl substituted analogue Sn[HiLPr]2 .The molecular structures of the title compounds iPr][HiLPr] (2) is structurally closely related to 1, as the Sn atom exhibits a trigonal\u2013bipyramidal coordination with the two amidinate N atoms in the axial positions . The lone pair at Sn is formally replaced by a chlorido ligand, which is almost perpendicular to the Sn\u2014N bonds . The Sn\u2014C bond lengths are 2.160\u2005(3)\u2005\u00c5 (Sn\u2014C2) and 2.154\u2005(3)\u2005\u00c5 (Sn\u2014C11) and therefore considerably shorter than in compound 1. This finding can be traced back to the higher oxidation number of the Sn atom. Similar Sn\u2014C bond lengths have been observed in the related tin(IV) derivatives SnCl3[HiLPr] [Sn\u2014C 2.132\u2005(3)\u2005\u00c5] and SnCl2[iLPr](THF) \u2212 ligands and deprotonated [iLPr]2\u2013 ligands are virtually equal, the Sn\u2014C(carborane) bond is obviously not noteworthily influenced by the bonding situation within the amidinate moiety. In contrast, in 2 the Sn\u2014N contact to the deprotonated carboranylamidinate ligand is considerably shorter than that to the protonated ligand . As has been discussed previously, the M\u2014N contact in [iLPr]2\u2013 complexes can be regarded as a distinct single bond, while the M\u2014N contact in [HiLPr]\u2212 complexes should be described as a secondary coordinative inter\u00adaction \u2005\u00c5, SnCl2[iLPr](THF): Sn\u2014N 205.0\u2005(2)\u2005\u00c5; Harmgarth et al., 2017The tin(IV) complex SnCl are clearly in the range of double bonds, while the C\u2014N bonds to the protonated N atoms can be regarded as single bonds. These values are consistent with those observed in other tin complexes with [HLR]\u2212 ligands (e.g. Sn[HLiPr]2: C=N(Sn) 1.287\u2005(3), C\u2014N(H) 1.347\u2005(4)\u2005\u00c5; Dr\u00f6se et al., 20103[HiLPr]: C=N(Sn) 1.288\u2005(4), C\u2014N(H) 1.339\u2005(4)\u2005\u00c5; Harmgarth et al. 20172, however the difference is less pronounced in this case . The \u03c0 electron density within the amidinate group in the [iLPr]2\u2013 ligand in 2 is clearly inverted, resulting in a long C\u2014N(Sn) bond [C3\u2014N1 1.372\u2005(4)\u2005\u00c5] and a short C=N bond to the non-coordinated N atom [C3\u2014N2 1.269\u2005(5)\u2005\u00c5]. Similar values have been obtained in SnCl2[iLPr](THF) \u2212 and [RL]2\u2013 ligands always display a typical \u03baC,\u03baN-chelating mode, this may explain why the formation of the complex SnCl[iLPr][HiLPr] (2) is preferred over a homoleptic complex Sn[iLPr]2, where the Sn atom would be tetra\u00adhedrally coordinated. Nonetheless, the example of SnCl2[iLPr](THF) (solv. = solvent) might exist.In summarizing the results reported here, in tin carboranylamidinates a trigonal\u2013bipyramidal \u2005\u00c5] and between isopropyl groups in 2 [C5\u22efC15 3.670\u2005(7)\u2005\u00c5], respectively. In both compounds, the free N\u2014H groups are not involved in hydrogen bonding.In both aFor reviews on amidinate complexes, see: Collins 2011\u2019 Edelmanet al. spectrometer in THF-d8 at 295\u2005(2)\u2005K. 1H and 13C NMR shifts are referenced to Si(CH3)4, 119Sn shifts to Sn(CH3)4 (each \u03b4 = 0 ppm). IR spectra were measured on a Bruker Vertex V70 spectrometer equipped with a diamond ATR unit, electron impact mass spectra on a MAT95 spectrometer with an ionization energy of 70\u2005eV. Elemental analyses were performed using a VARIO EL cube apparatus.All operations were performed under an argon atmosphere using standard Schlenk techniques. THF and toluene were distilled from sodium/benzo\u00adphenone under argon. NMR spectra were recorded on a Bruker DPX400 (Cy]2 (1): oPreparation of Sn was treated in situ with SnCl2 and then stirred for 24\u2005h. The solvent was subsequently removed in vacuo, the solid residue extracted with toluene (40\u2005mL) and the insoluble matter filtered off. The filtrate was concentrated to a total volume of ca 10\u2005mL. Cooling to 278\u2005K for 2\u2005d afforded colorless, block-like single-crystals of 1. Yield: 0.58\u2005g (20%). Analysis calculated for C30H66B20N4Sn, M = 817.81\u2005g\u2005mol\u22121: C 44.06, H 8.13, N 6.85%. Found: 43.66, H 8.00, N 6.25%. IR: \u03bd 3423 w (\u03bd NH), 2935 m (\u03bd CH2), 2854 m (\u03bd CH2), 2560 m (\u03bd BH), 1623 m (\u03bd C=N), 1448 m (\u03b4 CH2) cm\u22121. 1H NMR \u2005K): \u03b4 4.50 , 3.24 , 1.70\u20132.80 , 1.08\u20131.98 ppm. 13C NMR \u2005K): \u03b4 143.0 (NCN), 80.3 (C\u2014Sn), 77.0 (C\u2014CN2), 54.6 (CH Cy), 34.8 (CH2 Cy), 25.7 (CH2 Cy), 25.4 (CH2 Cy), 25.1 (CH2 Cy), 25.0 (CH2 Cy) ppm. 119Sn NMR \u2005K): \u03b4 \u221246 ppm. MS: m/z (%) 818\u2005(27) [M]+, 467\u2005(19) [C15H32B10N2Sn]+, 350\u2005(24) [C15H32B10N2]+, 269\u2005(100) [C9H23B10N2]+, 143\u2005(54) [C2H10B10]+.iPr] (2):Preparation of SnCl[L][SnCl5(THF)]. Yield: 2.13\u2005g. In addition to the IR data reported by Junk & Leary (20007Li and 119Sn) data. 7Li NMR \u2005K): \u03b4 = \u22120.76 ppm. 119Sn NMR \u2005K): \u03b4 = \u2212641 ppm. Concentration of the mother liquid to ca. 10\u2005ml followed by cooling again to 278\u2005K for several days afforded 0.13\u2005g (5%) of 2 as colorless, plate-like single-crystals. Analysis calculated for C18H49B20ClN4Sn, M = 692.00\u2005g\u2005mol\u22121: C 31.24, H 7.14, N 5.12%. Found: 31.02, H 6.88, N 4.98%. IR: \u03bd 3410 w (\u03bd NH), 2964 m, 2930 w , 2616 m, 2569 s (\u03bd BH), 1628 m, 1596 vs (\u03bd C=N), 1461 m, 1372 m cm\u22121. 1H NMR \u2005K): \u03b4 5.56 , 4.33\u20134.39 , 3.30\u20133.42 , 1.7\u20133.2 , 1.23\u20131.56 , 0.96\u20131.05 ppm. 13C NMR \u2005K): \u03b4 140.6 (NCN), 139.1 (NCN), 74.4 (C\u2014CN2), 73.2 (C\u2014CN2), 69.8 (C\u2014Sn), 52.0 (CH iPr), 48.3 (CH iPr), 25.5 (CH3), 23.1 (CH3) ppm. 119Sn NMR \u2005K): \u03b4 290 ppm. MS: m/z (%) 692\u2005(49) [M]+, 656\u2005(76) [C18H49B20N4Sn]+, 423\u2005(48) [C9H24B10ClN2Sn]+, 388\u2005(82) [C9H24B10N2Sn]+, 211\u2005(100) [C5H13B10N2]+, 171\u2005(18) [C3H10B10N]+.eary 2000, the com3 groups in 2 were allowed to rotate freely around the C\u2014C vector, the corresponding C\u2014H distances were constrained to 0.98\u2005\u00c5. C\u2014H distances within CH2 groups were constrained to 0.99\u2005\u00c5, C\u2014H distances within CH groups to 1.00\u2005\u00c5. H atoms attached to B and N atoms were located in the difference-Fourier map, B\u2014H distances were restrained to 1.12\u2005(2)\u2005\u00c5 and N\u2014H distances to 0.88\u2005(2)\u2005\u00c5. The Uiso(H) values were set at 1.5Ueq(C) for the methyl groups in 2, and at 1.2Ueq(X) in all other cases. For 1, the reflections (100) and compound_1, compound_2. DOI: 10.1107/S2056989017012671/zl2714compound_1sup2.hklStructure factors: contains datablock(s) compound_1. DOI: 10.1107/S2056989017012671/zl2714compound_2sup3.hklStructure factors: contains datablock(s) compound_2. DOI: 1565456, 1565455CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atoms are bridged by two carboxyl\u00adate anions.The crystal structure of a dinuclear tetra\u00adcarboxyl\u00adate complex of manganese(II) is reported wherein the Mn 2(C7H4NO4)4(C6H7N)4] or [Mn2(\u03bc-NBz)2(\u03ba2-NBz)2(4-Mepy)4], where NBz is 4-nitro\u00adbenzoate and 4-Mepy is 4-methyl\u00adpyridine, is a centrosymmetric dinuclear complex in which the MnII atoms are bridged by two NBz ligands with Mn\u22efMn = 4.1324\u2005(4)\u2005\u00c5. The MnII atom in this dimeric species is present in a distorted octa\u00adhedral environment with the four coordinating O atoms forming the equatorial plane and the two pyridyl N atoms occupying the axial sites. An important structural feature of the dimeric complex is that each of the bridging carboxyl\u00adate ligands binds to the metal ions in an asymmetric fashion involving bent and linear Mn\u2014O\u2014C units. The crystal packing is consolidated by C\u2014H\u22efO and C\u2014H\u22ef\u03c0 interactions.The title compound, [Mn While the former are in the range 56.95\u2005(4)\u2013150.77\u2005(4)\u00b0, the latter are in the range 88.02\u2005(5)\u201394.59\u2005(5)\u00b0.The highly distorted nature of the MnOIn the title compound, the carboxyl \u2013COO and \u2013NOO planes of the chelating NBz anion deviate slightly from the phenyl ring plane, forming dihedral angles of 2.6\u2005(3) and 23.6\u2005(4)\u00b0, respectively. According to Kaduk 2000 and KaduI2 and \u2013CO2 groups of the NBz ligand act as hydrogen acceptors (Table\u00a02D\u22efA separations for these weak contacts are in the range of 3.161\u2005(2) to 3.369\u2005(2)\u2005\u00c5, the =98.0%) were used as purchased from Sigma\u2013Aldrich . Aceto\u00adnitrile (>=99.9%) was used as purchased from Fisher Scientific . A 1:2 molar ratio of piroxicam:gentisic acid was dissolved in aceto\u00adnitrile. The concentration of piroxicam in aceto\u00adnitrile was near saturation I. DOI: 10.1107/S2056989016017321/hb7625Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016017321/hb7625Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016017321/hb7625Isup4.cmlSupporting information file. DOI: 1512430CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Mol\u00adecules are connected into a three-dimensional architecture by O\u2014H\u22efO,N hydrogen bonding.A distorted octa\u00adhedral NS 6H12NOS2)2(C12H10N4)]2\u00b72H2O, comprises a CdII atom, two di\u00adthio\u00adcarbamate (dtc) anions, a monodentate 3-pyridine\u00adaldazine ligand and a lattice water mol\u00adecule. The binuclear mol\u00adecule is constructed by the application of inversion symmetry. One dtc ligand simultaneously chelates one cadmium atom and bridges the centrosymmetric mate, while the other dtc ligand is chelating only. This leads to a centrosymmetric [Cd(dtc)2]2 core to which are appended two 3-pyridine\u00adaldazine ligands. The resulting NS5 donor set is based on an octa\u00adhedron. The three-dimensional mol\u00adecular packing is sustained by hydroxyl-O\u2014H(hydrox\u00adyl) and water-O\u2014H\u22efO(hydrox\u00adyl) hydrogen bonding, leading to supra\u00admolecular layers parallel to (101) which are connected by water-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonding; additional C\u2014H\u22efO, S \u03c0(chelate ring) inter\u00adactions are also evident. The retention of the central [Cd(dtc)2]2 core upon adduct formation is unprecedented in the structural chemistry of the zinc-triad di\u00adthio\u00adcarbamates.The asymmetric unit in the title binuclear compound, [Cd(C The overwhelming majority of structures are binuclear, [Cd(S2CNRR\u2032)2]2, arising from equal numbers of \u03bc2-tridentate and bidentate (chelating) ligands furan-2-ylmeth\u00adyl]2}3 2]n CH2CH2OH]2}n CH2CH(OMe)2]2}n CH2CH2OH]2}n species 2 is reacted with bases, e.g. pyridyl-donors, the original aggregate is disrupted in that no dtc links are retained between cadmium atoms. Thus, when archetypal, binuclear [Cd(S2CNEt2)2]2 ethyl\u00adene ethane CH2CH2OH)]2}2 and 3-pyridine\u00adaldazine were isolated and shown by X-ray crystallography that despite having one potentially bidentate bi-pyridyl ligand per Cd[S2CN(iPr)CH2CH2OH)]2 unit, the central binuclear core CH2CH2OH)]2 entity, a 3-pyridine\u00adaldazine ligand and one water mol\u00adecule of solvation. One di\u00adthio\u00adcarbamate (dtc) ligand coordinates in a chelating mode forming very similar Cd\u2014S bond lengths, i.e. the difference between the Cd\u2014Sshort and Cd\u2014Slong bond lengths is only 0.033\u2005\u00c5; this equivalence is reflected in the equivalence in the associated C1\u2014S1, S2 bond lengths, Table\u00a01bridging bond lengths are close to being equal, differing by only 0.010\u2005\u00c5, and are longer by ca 0.1\u2005\u00c5 than the non-bridging Cd\u2014S4 bond length, Table\u00a01anti disposition about both imine bonds, i.e. C18=N4 = 1.283\u2005(3)\u2005\u00c5 and C19=N5 = 1.277\u2005(3)\u2005\u00c5; the central, azo bond is 1.415\u2005(2)\u2005\u00c5. The pyridyl-N atoms are also anti but there are twists in the 3-pyridine\u00adaldazine mol\u00adecule, as seen in the value of the dihedral angle between the two pyridyl rings of 22.78\u2005(12)\u00b0.The mol\u00adecular structure of the binuclear title compound, isolated as a dihydrate, is shown in Fig.\u00a01hy\u00addroxy\u2014H\u22efOhy\u00addroxy\u2014H\u22efOwater\u2014H}n chains are formed as shown in Fig.\u00a02a. The water mol\u00adecules also form water-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonds on either side of the supra\u00admolecular layers sustained by O\u2014H\u22efO hydrogen bonds, Fig.\u00a02b. The pendent pyridyl-N atoms of Fig.\u00a02b are coordinating to cadmium atoms of successive layers so that a three-dimensional architecture results. Globally, and as seen from Fig.\u00a032CN(iPr)CH2CH2OH)]2}2 and 3-pyridine\u00adaldazine with the key links between them being hydrogen and coordinate bonding. Within this framework stabilized primarily by hydrogen-bonding inter\u00adactions, there are some second tier inter\u00adactions worthy of comment inter\u00adactions are present angle is 178\u00b0, in the inter-layer region, Table\u00a02Significant O\u2014H\u22efO hydrogen bonding is found in the mol\u00adecular packing of the binuclear title compound as would be expected from the chemical composition. Thus, mol\u00adecules are assembled into layers approximately parallel to (101) by hy\u00addroxy-O\u2014H\u22efO(hydrox\u00adyl) and hy\u00addroxy-O\u2014H\u22efO(water) hydrogen bonds as detailed in Table\u00a02et al., 20162-bridging 3-pyridine\u00adaldazine such as in the two most relevant compounds to the present study, namely {Cd[S2P(O-iPr)2]2}n 2]2}n 2]22\u00b7H2O}n (OH2)2]\u00b72ClO4}n . A good number of zinc and mercury binary di\u00adthio\u00adcarbamates are also known to adopt related binuclear [M(S2CNRR\u2019)2]2 aggregates owing to the presence of equal numbers of \u03bc2-tridentate and chelating ligands 2(hmta)]2, where hmta is hexa\u00admethyl\u00adene\u00adtetra\u00admine, for which an analogous centrosymmetric core and NS5 donor set as in the title compound was observed 2]2 cores in the structural chemistry of metal xanthates CH2CH2OH)]2 and 3-pyridine\u00adaldazine were dissolved in 1-propanol (15\u2005ml). The solution was carefully covered with hexa\u00adnes. Yellow blocks were obtained via slow diffusion of hexa\u00adnes into the 1-propanol solution over two weeks. m.p. 389\u2013391\u2005K. IR (cm\u22121): 1449 (m) \u03bd(C=N), 1173 (s) \u03bd(C\u2014S). 1H NMR: \u03b4 9.04 , 8.81 , 8.72 , 8.29 , 7.56 , 5.22 , 4.83 , 3.74 , 3.68 , 1.18 . TGA: three steps, corresponding to loss of water , loss of the 3-pyridine\u00adaldazine ligand , and decomposition down to cadmium sulfide .Cd[SUiso(H) set at 1.2\u20131.5Ueq(C). The oxygen-bound H atoms were located in a difference Fourier map but were refined with a distance restraint of O\u2014H = 0.84\u00b10.01\u2005\u00c5, and with Uiso(H) set at 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016012214/wm5312sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016012214/wm5312Isup2.hklStructure factors: contains datablock(s) I. DOI: 1496352CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Acta Cryst. E71, o125-o126. The motivation for this redetermination follows from negligence of important features of the difference electron-density maps as well as from wrongly applied constraints, which significantly affect the structural model. The corrections affect mainly the positions of the H atoms involved in the hydrogen bonds (centered on the primary amine group for which the H atom turned out to be disordered over two positions about the centre of an N\u22efH\u22efO hydrogen bond) and the methyl group, which is disordered and has now been remodelled.The title structure, 4-amino\u00adbenzoic acid 4-methyl\u00adpyridine/4-methyl\u00adpyridinium 4-amino\u00adbenzoate 0.58/0.42, has been redetermined from the data published by Kumar 6H7N\u00b7C7H7NO2)\u00b70.42(C6H8N+\u00b7C7H6NO2\u2212), has been redetermined from the data published by Kumar et al. \u2005\u00c5], while the H atoms of the primary amine group account more realistically for the hydrogen-bond pattern after the removal of the positional constraints. All the O\u2014H\u22efN or N\u2014H\u22efO hydrogen bonds which are present in the title structure are of moderate strength.The title structure, 4-amino\u00adbenzoic acid 4-methyl\u00adpyridine/4-methyl\u00adpyridinium 4-amino\u00adbenzoate 0.58/0.42, 0.58(C al. 2015. Acta Cr Figs. 1et al. = 1.2Ueq(Nprimary/secondary amine).In the course of recalculation of suspect structures that were retrieved from the CSD, the structure determination of the title structure by Kumar al. 2015, CSD refet al., 2006et al. (H1x) and 0.42(7) (H1y) Figs. 1 and 2 \u25b8.x\u22efN2iii/O1\u22efH1yiii\u2014N2iii is quite long for an O\u22efN hydrogen bond with a disordered bridging hydrogen atom, i.e. for a hydrogen atom the substantial part of its electron density is situated along the connecting line between the donor/acceptor atoms as happens in O1\u2014H1x\u22efN2iii/O1\u22efH1yiii\u2014N2iii of the title structure x hydrogen trioxo\u00adfluoro\u00adphosphate(1\u2212)x monohydrate/2,4,6-tri\u00adamino\u00adpyrimidinium(2+)x)(1\u2013 trioxo\u00adfluoro\u00adphosphate(2\u2212)x) \u2212 pKa(acid) is close to 0 , or ionized, forming a salt x\u00a0+\u00a01, y, z\u00a0+\u00a01] with a bridging hydrogen atom disordered over two positions (H1x and H1yiii) forms a finite D(3) pattern \u2212x\u00a0+\u00a01, \u2212y, z\u00a0\u2212\u00a0a--N1\u2013H1b and atom O2iv /\u03c3(i) < 0.6}. The indicators of the refinement of such a model were substanti\u00adally higher: Robs = 0.0503, Rwobs = 0.1035, Rall = 0.0930, Rwall = 0.1119. The condition for the observed diffractions was I/\u03c3(I) > 3, cf. Table\u00a02It is worthwhile mentioning that the recalculation of the original model with 10.1107/S2056989017013226/su5390sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989017013226/su5390Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017013226/su5390Isup3.cmlSupporting information file. DOI: 1574686CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The packing of the title compound features short C\u2014I\u22efN contacts. 18H18I2N2O2, consists of one half-mol\u00adecule, completed by the application of inversion symmetry. The mol\u00adecule adopts the typical structure for this class of bis-benxozazines, characterized by an anti orientation of the two benzoxazine rings around the central C\u2014C bond. The oxazinic ring adopts a half-chair conformation. In the crystal, mol\u00adecules are linked by C\u2014I\u22efN short contacts [I\u22efN = 3.378\u2005(2)\u2005\u00c5], generating layers lying parallel to the bc plane.The asymmetric unit of the title compound, C So, an iodine functional bis-1,3-benzoxazine, namely 3,3\u2032-bis\u00ad has been synthesized in high yield and purity.As the electrophilic character of the substituents affects the stability both of the reaction inter\u00admediates and the benzoxazine ring bond \u2005\u00c5, \u03b8 =129.6\u2005(2)\u00b0, \u03c6 = 283.6\u2005(3)\u00b0: with respect to the plane formed by O1/C3/C4/C5, the deviations of C2 and N1 are 0.301\u2005(3) and \u22120.320\u2005(3)\u2005\u00c5, respectively. The observed C\u2014O bond length [1.376\u2005(3)\u2005\u00c5] is in a good agreement with the related p-fluoro and p-bromo structures . Moreover, the C2\u2014N1\u2014C1 [112.6\u2005(2)\u00b0] and C5\u2014N1\u2014C1 [113.0\u2005(2)\u00b0] angles are larger than the mean value of sp3 hybridization in ammonia , monohydrate set to 1.2Ueq of the parent atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017005047/hb7668sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017005047/hb7668Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017005047/hb7668Isup3.cmlSupporting information file. DOI: 1541561CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atoms in {Zn[S2CN(R)CH2CH2OH]2(pyridine)\u00b7pyridine}, for R = Me (I) and Et (II), are coordinated non-symmetrically by two di\u00adthio\u00adcarbamate ligands and by a pyridine ligand, resulting in an NS4 donor set that defines a distorted geometry in each case; the non-coordinating pyridine mol\u00adecules are connected with the Zn-containing mol\u00adecules via O\u2014H\u22efN(pyridine) hydrogen bonds. The mol\u00adecular packing features significant (hy\u00addroxy)O\u2014H\u22efO(hy\u00addroxy) hydrogen bonding, in each case leading to supra\u00admolecular chains with zigzag (I) or helical (II) arrangements.The Zn 4H8NOS2)2(C5H5N)]\u00b7C5H5N (I) and [Zn(C5H10NOS2)2(C5H5N)]\u00b7C5H5N (II), which differ by having di\u00adthio\u00adcarbamate N-bound methyl (I) and ethyl (II) groups, is the coordination of each ZnII atom by two non-symmetrically chelating di\u00adthio\u00adcarbamate ligands and by a pyridine ligand; in each case, the non-coordinating pyridine mol\u00adecule is connected to the Zn-containing mol\u00adecule via a (hy\u00addroxy)O\u2014H\u22efN(pyridine) hydrogen bond. The resulting NS4 coordination geometry is closer to a square-pyramid than a trigonal bipyramid in the case of (I), but almost inter\u00admediate between the two extremes in (II). The mol\u00adecular packing features (hy\u00addroxy)O\u2014H\u22efO(hy\u00addroxy) hydrogen bonds, leading to supra\u00admolecular chains with a zigzag arrangement along [10-1] (I) or a helical arrangement along [010] (II). In (I), \u03c0\u2013\u03c0 [inter-centroid distances = 3.4738\u2005(10) and 3.4848\u2005(10)\u2005\u00c5] between coordinating and non-coordinating pyridine mol\u00adecules lead to stacks comprising alternating rings along the a axis. In (II), weaker \u03c0\u2013\u03c0 contacts occur between centrosymmetrically related pairs of coordinating pyridine mol\u00adecules [inter-centroid separation = 3.9815\u2005(14)\u2005\u00c5]. Further inter\u00adactions, including C\u2014H\u22ef\u03c0(chelate) inter\u00adactions in (I), lead to a three-dimensional architecture in each case.The common structural feature of the title compounds, [Zn(C While many similarities are apparent in their coordination propensities ethyl\u00adene (bpe). With Zn(S2COEt)2, a 1:1 compound can be prepared which crystallography shows to be a one-dimensional coordination polymer with a zigzag arrangement group only the dimeric aggregate [Zn(S2COCy)2]2(bpe) could be isolated 2]2, in that 1:1 one-dimensional coordination polymers can be formed with bpe when R = i-Pr di\u00adthio\u00adcarbamates of bpe, where only binuclear species, [Zn(S2CNR2)2]2(bpe), have been isolated, e.g. R = Me CH2CH2OH 2]2, where the dimeric aggregate, mediated by Zn\u2014S bridges, self-assembles into a three-dimensional architecture based on hydrogen bonding CH2CH2OH]2}2(bipy), which allows for the construction of a doubly inter\u00adpenetrated architecture. When the bridge is pyrazine, the three-dimensional architecture is sustained by (hy\u00addroxy)O\u2014H\u22efO(hy\u00addroxy) hydrogen bonding exclusively CH2N(H)C(=O)C(=O)N(H)CH2(3-pyrid\u00adyl), i.e. LH2, the dimeric {Zn[S2CN(Me)CH2CH2OH)2]2}2(LH2) aggregates are inter\u00adwoven into supra\u00admolecular chains sustained by hydrogen bonding CH2CH2OH]2(pyridine)\u00b7pyridine} for R = Me (I)In order to overcome the reluctance of zinc(II) di\u00adthio\u00adcarbamates to generate extended supra\u00admolecular architectures, the di\u00adthio\u00adcarbamate ligands can be functionalized with hydrogen-bonding potential, 2CN(R)CH2CH2OH]2(pyridine)\u00b7.pyridine}, for R = Me (I)long - Zn\u2014Sshort. For the S1-di\u00adthio\u00adcarbamate ligand, \u0394Zn\u2014S = 0.23\u2005\u00c5, but this decreases to 0.17\u2005\u00c5 for the S3-ligand. From the data in Table\u00a01\u03c4, which ranges from 0.0 to 1.0\u00b0 for ideal square-pyramidal to trigonal-bipyramidal, respectively \u03c4 is 0.56, indicating a small tendency towards trigonal\u2013bipyramidal, certainly when compared with the coordination geometry for (I)i.e. 166.375\u2005(19)\u00b0. As for (I)etc. The solvent mol\u00adecule in (II)via a hydrogen bond.To a first approximation the structure of (II)2CN(R)CH2CH2OH]2 mol\u00adecules in (I)Despite the relatively close agreement between the coord\u00adination geometries of the Zn. A view of the unit-cell contents is shown in Fig.\u00a04b. As for (I)In the crystal of (II)e.g. as anti-cancer agents 2(pyridine), which was motivated by the desire to destroy the binuclear structure observed for the binary di\u00adthio\u00adcarbamate compound to form a lighter (i.e. lower mol\u00adecular weight) species to facilitate chemical vapour deposition studies 2(pyridine) species. More sophisticated monodentate nitro\u00adgen-donor adducts are also known, such as substituted pyridines, e.g. 3-hy\u00addroxy\u00adpyridine 2CNEt2)2(pyridine) adduct has also been characterized as a mono-pyridine solvate CH2CH2OH]2, R = Me and Et, precursors were prepared as per established procedures CH2CH2OH]2, R = Me and Et (50\u2005mg), was dissolved in pyridine (10\u2005ml) and carefully layered with hexa\u00adnes (10\u2005ml). Crystals were harvested directly from solution and mounted immediately onto the diffractometer to avoid loss of pyridine.The Zn[SUiso(H) set to 1.2\u20131.5Ueq(C). The O-bound H atoms were located in difference-Fourier maps but were refined with a distance restraint of O\u2014H = 0.84\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.5Ueq(O). For (I)\u22123, respectively, were located 1.03 and 1.02\u2005\u00c5 from the Zn atom. For (II)\u22123, respectively, were located 1.02 and 0.61\u2005\u00c5 from the Zn atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989017010568/wm5406sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989017010568/wm5406Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017010568/wm5406IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1562767, 1562766CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The bridging mode of the organic ligands leads to the formation of a polymeric layer structure parallel to the ab plane.The zinc cation in the structure has a N 8H4O6S)(C10H8N2S)(H2O)]\u00b70.26H2O}n, is characterized by a layered arrangement parallel to the ab plane. The zinc cation is five-coordinated in a slightly distorted trigonal\u2013bipyramidal coordination environment defined by two pyridine ligands, two carboxyl\u00adate groups of two thio\u00adphene di\u00adcarboxyl\u00adate ligands, and by one water mol\u00adecule. The ethyl\u00adene bridge in the dioxine ligand is disordered over two sets of sites [occupancy ratio 0.624\u2005(9):0.376\u2005(9)]. Several hydrogen-bonding inter\u00adactions of the types O\u2014H\u22efO, C\u2014H\u22efO, C\u2014H\u22efS and C\u2014H\u22efN ensure the cohesion within the crystal structure.The crystal structure of the title polymer, {[Zn(C The equatorial Zn1\u2014N1 bond length is 2.1131\u2005(18)\u2005\u00c5, while the axial Zn1\u2014N2 bond is longer, 2.2107\u2005(18)\u2005\u00c5. Similarly, the two equatorial Zn1\u2014O bond lengths, ranging from 1.9835\u2005(15) to 2.0285\u2005(15)\u2005\u00c5, are shorter than the axial Zn1\u2014O7 bond of 2.1375\u2005(17)\u2005\u00c5. These are typical values, numerical details of which are given in Table\u00a01In the crystal structure of (1), the zinc ion is coordinated by four organic ligands and one water mol\u00adecule, giving rise to a slightly distorted trigonal\u2013bipyramidal coordination environment. Two nitro\u00adgen atoms are delivered by two symmetry-related pyridine ligands, two oxygen atoms of two carboxyl groups stem from two symmetry-related thio\u00adphene carboxyl\u00adate ligands, and one O atom from the aqua ligand Fig.\u00a01. In the ab plane to the coordinating O5 atom of the carboxyl group. Another (pyridine)C\u2014H group (C18\u2014H18A) is hydrogen-bonded to the disordered O8 atom of the lattice water mol\u00adecule. Three O\u2014H\u22efO inter\u00adactions are present between the coordinating water mol\u00adecule to either the carboxyl group oxygen atoms or the dioxine oxygen atom in the thio\u00adphene derivative with D\u22efA distances ranging between 2.733\u2005(2) and 3.123\u2005(2)\u2005\u00c5 and corresponding O\u2014H\u22efO angles of 135\u2005(2) and 159\u2005(2)\u00b0. Numerous other C\u2014H\u22efO inter\u00adactions are present between the disordered dioxine C\u2014H groups and a carboxyl O atom (O6) or the lattice water atom O8. Other C\u2014H\u22efO inter\u00adactions involve pyridyl C\u2014H groups and the carboxyl O3 atom. In addition, one C\u2014H\u22efS inter\u00adaction and one C\u2014H\u22efN inter\u00adaction are found between pyridyl C\u2014H groups and the sulfane S1 atom or the pyridyl N1 atom C\u2014H group was prepared as reported , H2ttdc , 4,4\u2032-di\u00adthiodi\u00adpyridine , 5\u2005ml di\u00admethyl\u00adformamide and 3\u2005ml water was mixed and heated at 353\u2005K for 3 days. After cooling, 0.17\u2005g light-yellow crystals were collected in a yield of 32%.2,3-Di\u00adhydro\u00adthieno could not be retrieved from difference maps and thus were not part of the model. Two carbon atoms of the dioxine moiety are disordered over two sets of sites and were refined in two parts (C3\u2013C4/C3A\u2013C4A) with a refined occupancy ratio of 0.624\u2005(9)/0.376\u2005(9). Soft restraints were applied on the disordered atoms to keep their geometries and atomic displacement parameters reasonable.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017002031/wm5363sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017002031/wm5363Isup2.hklStructure factors: contains datablock(s) I. DOI: 1528425CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In this paper, we report the discovery of a \u201creciprocal\u201d carbonyl-carbonyl interaction with substantial back and forth n\u2192\u03c0* and \u03c0\u2192\u03c0* electron delocalization between neighboring carbonyl groups. We have carried out experimental studies, analyses of crystallographic databases and theoretical calculations to show the presence of this interaction in both small molecules and proteins. In proteins, these interactions are primarily found in polyproline II (PPII) helices. As PPII are the most abundant secondary structures in unfolded proteins, we propose that these local interactions may have implications in protein folding.Carbonyl-carbonyl \u03c0* non covalent interactions affect the structure and stability of small molecules and proteins. Here, the authors carry out experimental studies, analyses of crystallographic databases and theoretical calculations to describe an additional type of carbonyl-carbonyl interaction.Carbonyl-carbonyl Intermolecular noncovalent interactions of varying magnitude are also responsible for the existence of different states of matter4. Carbonyl-carbonyl (C\u2550O\u00b7\u00b7\u00b7C\u2550O) n\u2192\u03c0* interactions where one of the lone pairs (n) on the oxygen atom of a carbonyl group is delocalized over the antibonding \u03c0* orbital of a nearby carbonyl C\u2550O bond (\u03c0*C\u2550O) along the B\u00fcrgi-Dunitz trajectory5 (\u2220O\u00b7\u00b7\u00b7C\u2009\u2550\u2009O\u2009~\u2009109\u00b0) have attracted a great deal of attention in recent years11. Previous studies have shown that C\u2550O\u00b7\u00b7\u00b7C\u2550O n\u2192\u03c0* interactions not only influence geometries of important small molecules15 but also play crucial roles in determining the three dimensional structures of polyesters16, peptides17, peptoids21 and proteins25. C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions between the side-chain and backbone carbonyl groups of Asp, Asn, Glu, and Gln were also observed in the high-resolution crystal structures of proteins27. C\u2550O\u00b7\u00b7\u00b7C\u2550O n\u2192\u03c0* interaction is characterized by a short O\u00b7\u00b7\u00b7C\u2550O distance (d) of less than 3.22\u2009\u00c5 [the sum of van der Waals radii of carbon and oxygen atom28], bond angle \u2220O\u00b7\u00b7\u00b7C\u2550O (\u03b8) of ~109\u00b0 and the pyramidality of the acceptor carbon atom towards the donor oxygen atom29. Direct spectroscopic evidence for n\u2192\u03c0* interaction was recently reported by using gas-phase infrared spectroscopy30.Nature effectively uses combinations of weak noncovalent interactions in the functional forms of various biologically important molecules such as nucleic acids and proteinsn\u2192\u03c0* interaction both donor and acceptor C\u2550O bonds will be polarized, which will make the acceptor carbonyl oxygen atom a better electron donor and the donor carbonyl carbon atom a better electron acceptor. The acceptor carbonyl oxygen, therefore, can donate electrons to another nearby carbonyl carbon either to form a sequential chain of O\u00b7\u00b7\u00b7C contacts 16 and proteins22, reciprocal n\u2192\u03c0* interactions remained unexplored. Allen and coworkers reported anti-parallel arrangements of carbonyl groups in ketone dimers that were bound together by two intermolecular C\u2550O\u00b7\u00b7\u00b7C\u2550O short contacts of dipolar nature31. Maccallum et al reported a similar geometrical arrangement of carbonyl groups in right-twisted \u03b2-strands and observed two chemically distinct dipolar C\u2550O\u00b7\u00b7\u00b7C\u2550O short contacts32. However, these C\u2550O\u00b7\u00b7\u00b7C\u2550O short contacts were considerably longer than the sum of van der Waals radii of C and O atoms.We anticipated that due to cts Fig.\u00a0 or it caons Fig.\u00a0. Althougn\u2192\u03c0* interactions should lead to back and forth donations between the carbonyl pairs. Based on our hypothesis, we discovered the presence of \u201creciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions\u201d both in small molecules and proteins. To establish the existence of reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions, we designed and synthesized model compounds and carried out X-ray crystallographic and theoretical studies. Further, we carried out Cambridge Structural Database (CSD)33 and Protein Data Bank (PDB)34 analyses to show that these interactions are widely present in small molecules and proteins. In proteins, these interactions are primarily found in random coils and turn regions. Based on our observations we propose that reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions may be a key local interaction that restricts the number of conformers of unfolded proteins and may have a role in protein folding.In this paper, we hypothesized that the polarization of the carbonyl groups by n\u2192\u03c0* interactions, we have synthesized N,N\u02b9-diacylhydrazines 1-8 having various substituents on either side of the carbonyl groups crystallized in nonplanar form with the carbonyl groups oriented almost orthogonal to each other , the two carbonyl groups stay far apart. The crystallographic distances d1 and d2 show no evidence of n\u2192\u03c0* interaction , an increase in the acceptor ability of the carbonyl CO-II is expected, which should increase n\u2192\u03c0* interaction from the oxygen atom of CO-I to the \u03c0*C\u2550O orbital of CO-II. However, the inductive electron withdrawal of Cl can be negated by the electron donation from the Cl lone pairs into the antibonding orbitals (\u03c3* and \u03c0*) of the adjacent carbonyl group (CO-II). In compound 4, we observed such electron delocalizations from the Cl lone pairs to both \u03c3* and \u03c0* orbitals of the C=O bonds of CO-II group, which contributed 0.67\u2009kcal\u2009mol\u22121 to the stabilization and presence of NBO second order perturbation energy . As can be anticipated, the nature of the two atoms/groups between the interacting carbonyl groups plays a key role in keeping the two carbonyl groups non coplanar and provides them the conformation required for reciprocal interactions th amino acid residue is defined as d1. The distance between the carbonyl oxygen of (i\u2009+\u20091)th residue and carbonyl carbon of ith residue is defined as d2. The corresponding \u2220O\u00b7\u00b7\u00b7C\u2550O angles are defined as \u03b81 and \u03b82, respectively deviates significantly from the B\u00fcrgi-Dunitz trajectory studied here show that shorter distances d1 and d2\u2009\u2264\u20093.2\u2009\u00c5 fall within the tail of the full distribution less than 10%, out of which 2184 showed the presence of reciprocal interactions in them. The PDB protein structures ranked by the number of reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions present in them are included in Supplementary Data\u00a0nge Fig.\u00a0. The angory Fig.\u00a0. These o22, Bartlett et al reported one-sided n\u2192\u03c0* interactions with d\u2009\u2264\u20093.20\u2009\u00c5 and 99o\u2009\u2264\u2009\u03b8\u2009\u2264\u2009119o. As we have applied the same distance (d\u2009\u2264\u20093.20\u2009\u00c5) and resolution (<1.6\u2009\u00c5) criteria, the reciprocal interactions observed here for angles 99o\u2009\u2264\u2009\u03b81, \u03b82\u2009\u2264\u2009119o would be observed as one-sided n\u2192\u03c0* interactions by using the criteria of Bartlett et al. As can be seen from Fig.\u00a0\u03b81 and \u03b82 in the range of 99\u00b0\u2013119\u00b0 is a very small percentage (6.5%) of the total number of reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions that are being reported here. This indicates that reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions are novel and distinct from one-sided n\u2192\u03c0* interactions reported previously.In a previous study1 and d2\u2009\u2264\u20093.20\u2009\u00c5) that covers the complete range of observed \u2220O\u00b7\u00b7\u00b7C\u2009=\u2009O angle (\u03b8) (70\u2013110\u00b0) clearly showed the presence of reciprocal n\u2192\u03c0* interactions , \u03c0\u2192\u03c0* interactions were observed for \u03b81 and \u03b82 values <90\u00b0 also (Table\u00a0n\u2192\u03c0* and two \u03c0\u2192\u03c0* interactions) could be substantial to protein stabilization. Based on the NBO calculations at B3LYP/6-311\u2009+\u2009G level, we observed that reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions contribute 0.27\u20134.41\u2009kcal\u2009mol\u22121 to the stabilization of proteins per amino acid pair . Secondary structure analyses using Stride38 show that reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions have considerable abundance in random coils (~20%) and turn regions (10%) of proteins but negligible presence in \u03b1-helices (0.35%) : ]. We have confirmed this by plotting the \u03c6, \u03c8 angles of residues in the random coil regions having reciprocal interactions Table\u00a0. This isons Fig.\u00a0. This isn\u2192\u03c0* interactions are twisted of the central two amino acids do not participate in any inter-residue hydrogen bonding. Therefore, we assume that these residues may participate in local reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions either between themselves or with their other neighbors, which should compensate for the lack hydrogen bonding interactions in them. A careful examination of the orientations of the carbonyl groups in various common \u03b2-turns indicated that reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interaction may be feasible between the first and the second residues of type I\u02b9 and II \u03b2-turns due to the favorable orientations of the two carbonyl groups but likely to be unfavoured in type I and II\u02b9 \u03b2-turns. In fact, analysis of the 10 protein crystal structures discussed above show that most of the reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interaction pairs found in \u03b2-turns were type II, followed by type IV (Supplementary Table\u00a0n\u2192\u03c0* interactions in \u03b1-helices and \u03b2-sheets22 (Pro\u2009>\u2009Gly\u2009>\u2009Ala). Analysis of distribution of reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions among the amino acid pairs in various proteins reveals that Pro\u2013Pro is the most abundant pair show a strong correlation against the stabilization energies due to n\u2192\u03c0* interactions [NBO second order perturbation energies E1n\u2192\u03c0*)( and E2n\u2192\u03c0*)(] reported in Tables\u00a0En\u2192\u03c0*)( for n\u2192\u03c0* interactions decreases with an increase in the O\u00b7\u00b7\u00b7C (d) in synthetic molecules 1\u20138, molecules taken from CSD and interacting amino acid pairs obtained from PDB (Tables\u00a0n\u2192\u03c0* [En\u2192\u03c0*)(] and \u03c0\u2192\u03c0* [E\u03c0\u2192\u03c0*)(] interaction energies reported in Tables\u00a0n\u2192\u03c0* interactions increases charge on donor oxygen lone pair (nO) and depletes it on acceptor carbonyl \u03c0*C\u2550O orbital, which correlate well with the strength of O\u00b7\u00b7\u00b7C distances between two dipoles could be used to understand the dipolar nature of interaction between them. As we know, antiparallel (T\u2009~\u2009180\u00b0) dipoles attract and parallel dipoles (T\u2009~\u20090\u00b0) repel each other whereas two orthogonal dipoles (T\u2009~\u200990\u00b0) have net zero dipolar interaction. In case of reciprocal interaction, the C\u2009=\u2009O\u00b7\u00b7\u00b7C\u2009=\u2009O torsion angles show an orientational preference [C\u2550O\u00b7\u00b7\u00b7C\u2550O torsion angle falls in 60\u00b0 to 90\u00b0 (or \u221260\u00b0 to \u221290\u00b0) range] as a consequence of the simultaneous restrictions on d1 and d2 (\u22643.2\u2009\u00c5). However, the values of the C\u2550O\u00b7\u00b7\u00b7C\u2550O torsion angles (~90\u00b0) suggest that there would be almost net zero interaction between the dipoles, eliminating the possibility of strong dipolar interactions. Therefore, we conclude that orbital delocalization is the major driving force for the stabilization of reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions. An elaborate energy decomposition analysis may be required for the accurate deconvolution of various factors contributing to the stabilization of reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O short contacts.The nature of C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions has been debated in the literature. While some consider them on Figs.\u00a0. In Fig.B Tables\u00a0. The ovenO) and \u03c0*C\u2550O orbital is weak. This weak donation from the first carbonyl group to the second is compensated by a back donation from the second carbonyl group to the first. In many cases, reciprocal \u03c0\u2192\u03c0* interactions were also observed along with reciprocal n\u2192\u03c0* interactions and their overall contributions to the stabilization of molecules having reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O short contacts could be significant. In proteins, C\u2550O\u00b7\u00b7\u00b7C\u2550O n\u2192\u03c0* interactions are present in all types of secondary structures. While one-sided n\u2192\u03c0* interactions are prevalent in \u03b1-helices23, reciprocal interactions are abundant in PPII helices and turn regions. Prevalence of reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions in PPII helices and turn regions of proteins suggests a possible role for these interactions in protein folding. Further, the presence of reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions in distorted \u03b1-helices and twisted \u03b2-sheets suggests that these interactions could stabilize secondary structures that deviate from their regular geometries. The reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions present at the interface of two different types of secondary structures could also help in stabilizing the strained amino acid residues that are present at these interfaces. In future, it would be interesting to investigate the ability of amino acid pairs having high propensity to get involved in reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions to stabilize PPII helices and \u03b2-turns. It would also be interesting to investigate if some non-peptidic fragments obtained from the CSD search having strong reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions could be used to stabilize PPII conformation or design peptide-turns. Finally, an energy decomposition analysis would provide better understanding of the forces that contributes to the stabilization of reciprocal C\u2550O\u00b7\u00b7\u00b7C\u2550O interactions.We conclude that reciprocal carbonyl-carbonyl interactions exist both in small organic molecules and proteins. However, due to geometrical constraints associated with such interactions, the approach of the donor oxygen atoms to the acceptor carbon atoms deviates significantly from the B\u00fcrgi-Dunitz trajectory, and therefore, electron delocalization between the oxygen lone pair . Details of the crystallization conditions are given in Supplementary Table\u00a0Single crystals of compounds 1\u20138 were determined by measuring X-ray intensity data. Bruker D8Venture APEX 342 single crystal home source X-ray diffractometer equipped with CMOS PHOTON 100 detector and Monochromated microfocus sources Mo K\u03b1 radiation (\u03bb\u2009=\u20090.71073\u2009\u00c5) were used for data collection in phi (\u03d5) and omega (\u03c9) scan strategy at room temperature (298\u2009K). The data was processed using SAINT43 and absorption correction was done using SADABS44 implemented in APEX 3. For structure solution XSHELL program based on SHELX45 was used. The non-hydrogen atoms were refined anisotropically and located in successive difference Fourier syntheses. The hydrogen atoms were fixed to neutron bond length using appropriate HFIX commands. ORTEP diagrams of compounds 1\u20138 (CCDC 1486577- 1486584) is provided in Supplementary Fig.\u00a05 crystallized with a water molecule in the asymmetric unit. However, for clarity we have not shown the water molecule in its ORTEP diagram. Compound 7 has disorder at chlorine atom; the occupancy of disordered chlorine atom namely Cl1A and Cl1B was refined using the PART command. Similar ADP restraint SIMU46 and rigid bond restraint DELU46 was applied to stabilize the anisotropic refinement. SADI46 instruction was used to restrain the distance to equal. The anisotropic displacement parameter for disordered chlorine atom was fixed using EADP46 constraint.Single crystal structures of compound 33 (CSD version 5.21 Nov. 2015) using Conquest47 (version 1.18) program. The fragment chosen for the search is shown in Supplementary Fig.\u00a0X is indicative for any atom. Only unique matching fragments were taken and the fragment was chosen in such a way that there are at least two carbonyl groups irrespective of their nature. Distances d1 (O2\u2013C5) and d2 (O6\u2013C1) are restricted to \u22643.2\u2009\u00c5. Angles [O2\u2013C5\u2013O6 (\u03b81) and O6\u2013C1\u2013O2 (\u03b82)] and dihedral angles (C1\u2013O2\u2013C5\u2013O6 and C5\u2013O6\u2013C1\u2013O2) were printed without any restriction. Only crystalline, non-ionic and non-polymeric organic molecules having no disorder and error with R factor\u2009\u2264\u20095% having at least three covalent bond separations between the carbonyl groups were considered in this search.Intramolecular C\u2550O\u00b7\u00b7\u00b7C\u2550O noncovalent interactions were searched and structural data were retrieved from Cambridge Structural Database34 using a search criterion of resolution <1.6\u2009\u00c5 with redundancy (pairwise sequence identity) less than 10%, downloaded on 19 January 2016. Out of 2269 proteins, 2184 showed the reciprocal n\u2192\u03c0* interaction. For proteins existing in polymeric form or for proteins containing amino acids in more than one conformation, Chain A and conformation A were chosen, except for 57 proteins where chain A is absent. Distance d1 is defined as distance between the ith amide oxygen to the subsequent (i\u2009+\u20091)th amide carbon, while d2 is defined as distance between the (i\u2009+\u20091)th amide oxygen to the ith amide carbon .A subset of 2269 protein was culled out from RCSB PDB48. The Hartree-Fock (HF)49 and the hybrid Becke 3-Lee-Yang-Parr (B3LYP)51 exchange correlation functional with 6-311\u2009+\u2009G basis set were used for the calculations. Natural bond orbital (NBO)35 analyses were performed on the crystal geometries of the synthetic molecules and small organic molecules obtained from CSD search. For proteins, the coordinates of the interacting amino acid residue pair were extracted using PyMOL52. The \u03b1-carbons of the amino acid residues adjacent to N and C termini of the amino acid pair were also included, so as to mimic a dipeptide with N and C termini capped with N(CO)Me and (CO)NMe, respectively. Finally, hydrogen atoms were added to the structure using PyMOL ( and E\u03c0\u2192\u03c0*) level of theory.All the calculations were performed by using Gaussian09 suite of quantum chemistry programswww.ccdc.cam.ac.uk/.The authors declare that the data supporting the findings of this study are available within the paper and its Supplementary Information files, and also are available from the corresponding author upon reasonable request. X-ray crystallographic data for structures reported in this study have been deposited at the Cambridge Crystallographic Data Centre (CCDC), under deposition number CCDC 1486577-1486584. These data can be obtained free of charge from the CCDC via Supplementary InformationSupplementary Data 1Supplementary Data 2Supplementary Data 3Supplementary Data 4Supplementary Data 5Supplementary Data 6Supplementary Data 7Supplementary Data 8Supplementary Data 9Peer Review File"} +{"text": "Only one enanti\u00adomer was observed crystallographically, resulting from the combination of (S)-sec-butyl\u00adamine with l-tartaric acid.The title hydrated mol\u00adecular salt was prepared by deprotonation of enanti\u00adopure 4H12N+\u00b7C4H5O6\u2212\u00b7H2O, was prepared by deprotonation of enanti\u00adopure l-tartaric acid with racemic sec-butyl\u00adamine in water. Only one enanti\u00adomer was observed crystallographically, resulting from the combination of (S)-sec-butyl\u00adamine with l-tartaric acid. The sec-butyl\u00adammonium moiety is disordered over two conformations related by rotation around the CH\u2013CH2 bond; the refined occupancy ratio is 0.68\u2005(1):0.32\u2005(1). In the crystal, mol\u00adecules are linked through a network of O\u2014H\u22efO and N\u2014H\u22efO hydrogen-bonding inter\u00adactions, between the ammonium H atoms, the tartrate hy\u00addroxy H atoms, and the inter\u00adstitial water, forming a three-dimensional supra\u00admolecular structure.The title hydrated mol\u00adecular salt, C Since enanti\u00adomers have identical physical properties, they cannot be separated by standard physical means such as distillation, crystallization, or chromatography. One common method to overcome this issue is to convert the racemic compound into a mixture of diastereomers through reaction with an enanti\u00adopure component -sec-butyl ammonium cation, the l-tartrate anion, and one mol\u00adecule of water in the asymmetric unit. The Flack parameter [\u20132.7\u2005(8)] was not of use in determining the absolute configuration of the sec-butyl\u00adamine in the crystal. The absolute configuration of the (S)-sec-butyl ammonium cation is therefore based on the known absolute configuration of the l-tartaric acid used during compound preparation. The final structure is disordered, with the sec-butyl ammonium moiety taking on two different rotamers about the C2\u2013C3 axis [refined occupancy ratio is 0.68\u2005(1):0.32\u2005(1)]. The major component takes on a conformation where the C4 methyl group and N9 ammonium are in a gauche relationship , while the minor component places the C4A methyl group anti\u00adperiplanar to the N9A ammonium . The C\u2014C bond lengths in the amine and tartrate units average 1.523\u2005(11)\u2005\u00c5 [1.516\u2005(22)\u2005\u00c5 for the minor component of the disorder] and 1.532\u2005(5)\u2005\u00c5, respectively. The C\u2014N bonds of the two components of the disorder average 1.498\u2005(17)\u2005\u00c5. The tartrate C\u2014OH bonds average 1.411\u2005(4)\u2005\u00c5, while the C\u2014O bonds of the carboxyl moieties average 1.257\u2005(4)\u2005\u00c5 for the one involved in hydrogen bonding with the amine, and 1.258\u2005(4)\u2005\u00c5 for the other. An intra\u00admolecular hydrogen bond [2.00\u2005(3)\u2005\u00c5] occurs with O12 acting as a hydrogen-bond donor to O11.The mol\u00adecular structure of the title hydrated mol\u00adecular salt is shown in Fig.\u00a01ip Fig.\u00a01a, whileum Fig.\u00a01b. The CA atom of the sec-butyl ammonium cation acts as a hydrogen-bond donor to O11 of the tartrate anion [1.89\u2005(2)\u2005\u00c5], and the tartrate O13 donates a hydrogen bond to O16 of water [1.83\u2005(3)\u2005\u00c5]. The water in turn acts as a hydrogen-bond donor to O10 [2.01\u2005(3)\u2005\u00c5] and O15 [1.93\u2005(4)\u2005\u00c5] of two adjacent symmetry-related mol\u00adecules. Three additional hydrogen bonds are formed from N9, with N9\u2014H9B donating to O12 of an adjacent mol\u00adecule [1.97\u2005(3)\u2005\u00c5], and N9\u2014H9C donating to both O13 [2.16\u2005(4)\u2005\u00c5] and O15 [2.20\u2005(4)\u2005\u00c5] of a second adjacent mol\u00adecule. Finally, O14 donates a hydrogen bond to O10 of an additional symmetry-related mol\u00adecule [1.58\u2005(5)\u2005\u00c5]. A view of the crystal packing reveals the amine, tartrate, and water mol\u00adecules form columns when viewed down the c axis was added to 40\u2005ml of water and stirred to ensure homogeneity. While stirring, l-tartaric acid was slowly added. The solution was covered and allowed to stand at ambient temperature. After 24\u2005h, crystal formation was evident. The crystallization process was allowed to continue undisturbed for one week, at which point a crystal for diffraction analysis was selected directly from the reaction mixture without further purification or isolation. The crystals can be isolated by vacuum filtration to yield a white crystalline solid .The title compound was prepared Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. The sec-butyl ammonium moiety displays a twofold disorder arising from two different rotamers being present that is best described as a 0.68\u2005(1):0.32\u2005(1) ratio of the two possible conformations. In the final cycles of refinement SAME restraints were applied to the two components of the disordered sec-butyl ammonium moiety and DFIX restraints were applied to the N\u2014H bonds [N\u2014H = 0.91\u2005(2)\u2005\u00c5] and the ammonium H\u22efH distances [H\u22efH = 1.50\u2005(2)\u2005\u00c5], to improve the refinement and geometry.Crystal data, data collection, and structure refinement details are summarized in Table\u00a0210.1107/S2056989017005448/su5364sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989017005448/su5364Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017005448/su5364Isup3.cmlSupporting information file. DOI: 1543331CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two CPy ligands act as monodentate N(pyridine)-bonding ligands, while the two NB anions act as bidentate ligands through the carboxyl\u00adate O atoms. The centrosymmetric dinuclear complex is generated by application of inversion symmetry, whereby the CdII atoms are bridged by the carboxyl\u00adate O atoms of two symmetry-related NB anions, thus completing the distorted N2O5 penta\u00adgonal\u2013bipyramidal coordination sphere of each CdII atom. The benzene and pyridine rings are oriented at dihedral angles of 10.02\u2005(7) and 5.76\u2005(9)\u00b0, respectively. In the crystal, C\u2014H\u22efN hydrogen bonds link the mol\u00adecules, enclosing R22(26) ring motifs, in which they are further linked via C\u2014H\u22efO hydrogen bonds, resulting in a three-dimensional network. In addition, \u03c0\u2013\u03c0 stacking inter\u00adactions between parallel benzene rings and between parallel pyridine rings of adjacent mol\u00adecules , as well as a weak C\u2014H\u22ef\u03c0 inter\u00adaction, may further stabilize the crystal structure.The asymmetric unit of the title compound, [Cd The two CPy ligands are monodentate (through the pyridine N atoms), while both NB anions act as bidentate ligands through their carboxyl\u00adate O atoms and its symmetry-related counterpart . Hence, this carboxyl\u00adate group not only chelates to one CdII atom but also bridges two CdII atoms , one chelating NB anion (O1 and O2) and two 3-cyano\u00adpyridine (CPy) ligands (N3 and N5), resulting in a distorted penta\u00adgonal\u2013bipyramidal environment. The five carboxyl\u00adate O atoms of the three NB anions around the CdII atom form a distorted penta\u00adgonal arrangement, with an average Cd1\u2014O bond length of 2.42\u2005\u00c5 (Table\u00a01i separation in the binuclear mol\u00adecule is 3.9360\u2005(15)\u2005\u00c5 and is comparable to the corresponding M\u2014M distances in the structurally related transition metal(II) complexes . The metal atoms are bridged by two NA ligand N and O atoms in (III), (V) and (VI), while they are bridged by two carboxyl\u00adate O atoms in (IV).The asymmetric unit of the title complex contains one Cdms Fig.\u00a01. The cenms Fig.\u00a02. Thus, e\u00c5 Table\u00a01. The dis\u00c5 Table\u00a01 and 2 \u25b8.M\u2014O angles are 60.92\u2005(12)\u00b0 in (II), 53.50\u2005(14)\u00b0 in (IV), 57.61\u2005(8)\u00b0 in (V), and 54.22\u2005(4) and 53.32\u2005(5)\u00b0 in (VI). The dihedral angles between the planar carboxyl\u00adate groups (O1/O2/C1 and O5/O6/C8) and the adjacent benzene [A (C2\u2013C7) and B (C9\u2013C14)] rings in the title structure are 17.18\u2005(13) and 3.36\u2005(12)\u00b0, respectively, while the benzene (A and B) and pyridine [C (N3/C15\u2013C19) and D (N5/C21\u2013C25)] rings are oriented at dihedral angles of A/B = 10.02\u2005(7)\u00b0, A/C = 72.70\u2005(7)\u00b0, A/D = 74.72\u2005(7)\u00b0, B/C = 82.28\u2005(7)\u00b0, B/D = 84.54\u2005(8)\u00b0 and C/D = 5.76\u2005(9)\u00b0.The near equalities of the C1\u2014O1 [1.264\u2005(3)\u2005\u00c5], C1\u2014O2 [1.241\u2005(3)\u2005\u00c5], C8\u2014O5 [1.256\u2005(3)\u2005\u00c5] and C8\u2014O6 [1.253\u2005(3)\u2005\u00c5] bonds in the carboxyl\u00adate groups indicate delocalized bonding arrangements, rather than localized single and double bonds. The O1\u2014Cd1\u2014O2 and O5\u2014Cd1\u2014O6 bite angles are reduced to 54.33\u2005(5) and 53.47\u2005(5)\u00b0, respectively. The corresponding O\u2014cpy\u22efOc (cpy = cyano\u00adpyridine and c = carboxyl\u00adate) and C\u2014Hnb\u22efOc (nb = nitro\u00adbenzoate) hydrogen bonds hydrogen bonds (Table\u00a02Cg1\u2013Cg2i and Cg3\u2013Cg4ii may further stabilize the structure, with centroid\u2013centroid distances of 3.885\u2005(1) and 3.712\u2005(1)\u2005\u00c5, respectively. A weak C\u2014H\u22ef\u03c0 inter\u00adaction = 1.2Ueq(C). The maximum and minimum electron densities were found at 1.43 and 0.80\u2005\u00c5 from atoms O2 and Cd1, respectively.The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a034\u00b78H2O in H2O (50\u2005ml) and 3-cyano\u00adpyridine in H2O (50\u2005ml) with sodium 3-nitro\u00adbenzoate in H2O (100\u2005ml) at 333\u2005K. The mixture was filtered and set aside to crystallize at ambient temperature for one week, giving colourless single crystals.The title compound was prepared by the reaction of 3CdSO10.1107/S2056989017002675/wm5366sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017002675/wm5366Isup2.hklStructure factors: contains datablock(s) I. DOI: 1533101CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The Zn-containing units are connected into a one-dimensional coordination polymer (zigzag topology) propagating in the [010] direction, with one di\u00adthio\u00adcarbamate ligand bridging in a \u03bc2-\u03ba3 mode, employing one pyridyl N and both di\u00adthio\u00adcarbamate S atoms, while the other is \u03ba2-chelating. In each case, the resultant ZnNS4 coordination geometry approximates a square pyramid, with the pyridyl N atom in the apical position. In the crystal, the chains are linked into a three-dimensional architecture by methyl- and pyridyl-C\u2014H\u22efS, methyl\u00adene-C\u2014H\u22efN(pyrid\u00adyl) and pyridyl-C\u2014H\u22ef\u03c0(ZnS2C) inter\u00adactions. The connection between the chain and the 4-methyl\u00adpyridine solvent mol\u00adecule is of the type pyridyl-C\u2014H\u22efN(4-methyl\u00adpyridine).The title compound, {[Zn(C Thus, a two-dimensional architecture is found in centrosymmetric {Zn[S2CN(Et)CH2py]2 residues, shown in Fig.\u00a01i.e. \u0394(Zn\u2014S) = [Zn\u2014S(long) \u2212 Zn\u2014S(short)], for the S1-di\u00adthio\u00adcarbamate ligand of 0.32\u2005\u00c5 is greater than the value of 0.10\u2005\u00c5 for the S3-di\u00adthio\u00adcarbamate ligand. For the Zn2-mol\u00adecule, these differences diminish to 0.23 and 0.09\u2005\u00c5 for the S5- and S7-di\u00adthio\u00adcarbamate ligands, respectively. The similarity of the structures is emphasized in the overlay diagram of Fig.\u00a022-\u03ba3 mode, chelating one ZnII cation and simultaneously bridging another via the pyridyl N atom. It is noted that it is the di\u00adthio\u00adcarbamate ligand that forms the more equivalent Zn\u2014S bond lengths in each residue that forms the bridging inter\u00adactions. The resultant coordination geometry for each ZnII cation is based on an NS4 donor set.The asymmetric unit of (I)et al., 1984i.e. S1\u2013S4 (r.m.s. deviation = 0.1721\u2005\u00c5), in the direction of the pyridyl N6 atom. For the Zn2-mol\u00adecule, the deviation of the Zn2 cation from the S4 plane is 0.6018\u2005(6)\u2005\u00c5 and the r.m.s. deviation through the S5\u2013S8 atoms is 0.1273\u2005\u00c5.For five-coordinate species, the value computed for \u03c4 is a useful indicator of the adopted coordination geometry, with the values of \u03c4 ranging from 0 to 1 for ideal square-pyramidal and trigonal\u2013bipyramidal geometries, respectively 2CN(Et)CH2py]2\u00b73-methyl\u00adpyridine}n, which was also isolated from an experiment attempting to coordinate isomeric methyl\u00adpyridines to the heavy element \u2212[S2CN(Et)CH2py], found in (I)2CN(Et)CH2py]2\u00b73-methyl\u00adpyridine}n salt CH2py]2, for R = Me, nBu and Ph inter\u00adaction of 2.98\u2005\u00c5 between the two mol\u00adecules comprising the asymmetric unit. This result is consistent with surveys of diorganotin bis\u00ad(di\u00adthio\u00adcarbamate)s in general 2, a recent survey indicated that secondary bonding inter\u00adactions occur in only 10% of their crystal structures CH2py]2 (generated from the reaction of Zn(NO3)2\u00b7H2O and \u2212[S2CN(Et)CH2py]) from 4-picoline. Suitable single crystals formed upon slow evaporation of the solvent (m.p. 337\u2013339\u2005K).The title compound was isolated from the recrystallization of Zn{[SUiso(H) values set at 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017010179/hb7691sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017010179/hb7691Isup2.hklStructure factors: contains datablock(s) I. DOI: 1561011CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A distinctive O\u2014H\u22efN-based synthon is present. Cl\u22efCl and \u03c0\u2013\u03c0 stacking inter\u00adactions further stabilize the crystal structure, forming a two-dimensional network parallel to the bc plane.The asymmetric unit of the title compound, 2C In recent years, the physicochemical properties of active pharmaceutical ingredients have been improved widely with the use of co-crystals via O\u2014H\u22efN hydrogen-bonding inter\u00adactions between (O1\u2014H1) of the carboxyl group and the nitro\u00adgen (N1) of BPY \u2005\u00c5, C3\u2014Cl1\u22efCl1iii = 151.71\u2005(1)\u00b0; symmetry code: (iii) 1\u00a0\u2212\u00a0x, y, z] \u2005\u00c5; Cg1 is the centroid of the N1/C6/C7/C8/C9/C10 ring; symmetry code: (ii) 1\u00a0\u2212\u00a0x, 2\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z]. The perpendicular distance between two parallel mol\u00adecules is 3.4812\u2005(15)\u2005\u00c5. This weak inter\u00adaction holds the hydrogen-bonded chains together, supporting a two-dimensional supra\u00admolecular network parallel to the bc plane, as seen in Fig.\u00a023TPC and BPY are inter\u00adconnected Y Table\u00a01. This O\u2014et al., 2001N,N\u2032-dioxide-3-hy\u00addroxy-2-naphthoic acid (1/2) , 10\u2005ml of a hot methano\u00adlic solution of BPY was added. The resulting solution was warmed over a water bath for half an hour and then kept at room temperature for crystallization. After a week, clear yellow plates were obtained. The crystal used for X-ray diffraction data collection was cut from a larger crystal.Uiso(H) = 1.2Ueq(C). The carb\u00adoxy\u00adlic acid hydrogen atom was freely refined, including its isotropic displacement parameter.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016013724/hg5476sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016013724/hg5476Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016013724/hg5476Isup3.cmlSupporting information file. DOI: 1501060CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Its reaction with copper chloride lead to the formation of a copper(II) paddle-wheel structure.Tranexamic acid is an anti\u00adfibrinolytic amino acid that exists as a zwitterion [ trans-4-(amino\u00admeth\u00adyl)cyclo\u00adhexane-1-carb\u00adoxy\u00adlic acid], is an anti\u00adfibrinolytic amino acid that exists as a zwitterion [trans-4-(ammonio\u00admeth\u00adyl)cyclo\u00adhexane-1-carboxyl\u00adate] in the solid state. Its reaction with copper chloride leads to the formation of a compound with a copper(II) paddle-wheel structure that crystallizes as a hexa\u00adhydrate, [Cu2Cl2(C8H15NO2)4]2+\u00b72Cl\u2212\u00b76H2O. The asymmetric unit is composed of a copper(II) cation, two zwitterionic tranexamic acid units, a coordinating Cl\u2212 anion and a free Cl\u2212 anion, together with three water mol\u00adecules of crystallization. The whole structure is generated by inversion symmetry, with the Cu\u22efCu axle of the paddle-wheel dication being located about a center of symmetry. The cyclo\u00adhexane rings of the zwitterionic tranexamic acid units have chair conformations. The carboxyl\u00adate groups that bridge the two copper(II) cations are inclined to one another by 88.4\u2005(8)\u00b0. The copper(II) cation is ligated by four carboxyl\u00adate O atoms in the equatorial plane and by a Cl\u2212 ion in the axial position. Hence, it has a fivefold O4Cl coordination sphere with a perfect square-pyramidal geometry and a \u03c45 index of zero. In the crystal, the paddle-wheel dications are linked by a series of N\u2014H\u22efCl hydrogen bonds, involving the coordinating and free Cl\u2212 ions, forming a three-dimensional network. This network is strengthened by a series of N\u2014H\u22efOwater, Owater\u2014H\u22efCl and Owater\u2014H\u22efO hydrogen bonds.Tranexamic acid [systematic name: It has important anti\u00adfibrinolytic activity and is used extensively in both trauma and normal surgery to prevent excessive blood loss \u2005\u00c5; symmetry code (i): \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01] of the paddle-wheel being located about a center of symmetry. Selected bond lengths and angles in the paddle-wheel dication are given in Table\u00a01\u2212 ion, Cl1, in the axial position. The Cu\u2014O distances vary from 1.950\u2005(4) to 1.991\u2005(3)\u2005\u00c5, with a longer Cu1\u2014Cl1 axial distance of 2.499\u2005(1)\u2005\u00c5 (Table\u00a01i), has a perfect square-pyramidal coordination sphere with a \u03c45 index of 0.0 \u2005\u00c5, \u03b8 = 176.3\u2005(6)\u00b0, \u03c6 = 358\u2005(13)\u00b0, and for ring C10\u2013C15 are Q = 0.568\u2005(6)\u2005\u00c5, \u03b8 = 6.0\u2005(6)\u00b0, \u03c6 = 137\u2005(6)\u00b0. The carboxyl\u00adate groups (C1/O1/O2 and C9/O3/O4) are inclined to the mean planes of the four planar atoms of the respective cyclo\u00adhexane rings (C3/C4/C6/C7 and C11/C12/C14/C15) by 67.5\u2005(6) and 85.8\u2005(7)\u00b0, while they are inclined to one another by 88.4\u2005(8)\u00b0. The ammonio\u00admethyl units, C5/C8/N1 and C13/C16/N2, are inclined to the mean planes of the four planar atoms of the respective cyclo\u00adhexane rings (C3/C4/C6/C7 and C11/C12/C14/C15) by 34.9\u2005(6) and 47.5\u2005(6)\u00b0.The conformations of the two zwitterionic tranexamic acid units differ slightly. The cyclo\u00adhexane rings (C2\u2013C7 and C10\u2013C15) have chair conformations; puckering parameters for ring C2\u2013C7 are 3+ groups of the zwitterionic tranexamic acid units and the coordinating and free Cl\u2013 ions are linked by a series of N\u2014H\u22efCl hydrogen bonds forming a three-dimensional framework and the hydro\u00adchloride of the cis-isomer (CHCAHC) were reported in 1966 , viz. tranexamic acid is inclined to the mean plane of the four planar atoms of the cyclo\u00adhexane ring by 48.9\u2005(2)\u00b0, compared to 67.5\u2005(6) and 85.8\u2005(7)\u00b0 in the title compound. The plane of the ammonio\u00admethyl unit (Car\u2014C\u2014N) is inclined to the same mean plane of the four planar atoms of the cyclo\u00adhexane ring by 37.4\u2005(2)\u00b0, compared to 34.9\u2005(6) and 47.5\u2005(6)\u00b0 in the title compound. Hence, on complexation the cyclo\u00adhexane rings are rotated about the Ccarboxyl\u00adate\u2014Ccyclo\u00adhexa\u00adne bonds (C1\u2014C2 and C9\u2014C10), most probably to minimize steric hindrance.A search of the Cambridge Structural Database . The Cu\u22efCu distances vary from ca 2.63 to 2.84\u2005\u00c5, with the carboxyl\u00adate groups being inclined to one another by ca 84.65\u201390\u00b0, and the Cu\u2014Cl distances varying from ca 2.41 to 2.49\u2005\u00c5. The values observed for the title compound fall within these limits . In all 13 compounds the copper atoms have perfect square-pyramidal geometry, with \u03c45 = 0.0.In the CSD over 1500 copper(II) paddle-wheel structures have been deposited. There are only 13 compounds in which the axial position is occupied by a Cl2\u00b72H2O in 20\u2005ml of aceto\u00adnitrile at ambient temperature and the mixture was stirred for 30\u2005min. The green solution obtained was filtered and the filtrate kept undisturbed at room temperature for slow evaporation. After five days green plate-like crystals of the title compound were obtained.Tranexamic acid dissolved in 10\u2005ml of deionized water was added dropwise to a transparent blue solution of CuClUiso(H) = 1.5Ueq(O). The ammonium H atoms and the C-bound H atoms were included in calculated positions and treated as riding: N\u2014H = 0.91\u2005\u00c5, C-H = 0.99\u20131.00\u2005\u00c5 with Uiso(H) = 1.5Ueq(N-ammonium) and 1.2Ueq(C) for other H atoms. In the final difference-Fourier map the residual density peaks are located at a distance of 1.2 and 0.9\u2005\u00c5, respectively, from the copper atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017012543/wm5416sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989017012543/wm5416Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017012543/wm5416sup3.pdfCSD search of axially Cl- ligated Cu-Cu paddle-wheel structures. DOI: 1571897CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Interleukin 17-producing \u03b3\u03b4 T (\u03b3\u03b4T17) cells have unconventional trafficking characteristics, residing in mucocutaneous tissues but also homing into inflamed tissues via circulation. Despite being fundamental to \u03b3\u03b4T17-driven early protective immunity and exacerbation of autoimmunity and cancer, migratory cues controlling \u03b3\u03b4T17 cell positioning in barrier tissues and recruitment to inflammatory sites are still unclear. Here we show that \u03b3\u03b4T17 cells constitutively express chemokine receptors CCR6 and CCR2. While CCR6 recruits resting \u03b3\u03b4T17 cells to the dermis, CCR2 drives rapid \u03b3\u03b4T17 cell recruitment to inflamed tissues during autoimmunity, cancer and infection. Downregulation of CCR6 by IRF4 and BATF upon \u03b3\u03b4T17 activation is required for optimal recruitment of \u03b3\u03b4T17 cells to inflamed tissue by preventing their sequestration into uninflamed dermis. These findings establish a lymphocyte trafficking model whereby a hierarchy of homing signals is prioritized by dynamic receptor expression to drive both tissue surveillance and rapid recruitment of \u03b3\u03b4T17 cells to inflammatory lesions. IL-17-producing \u03b3\u03b4 T (\u03b3\u03b4T17) cells position in barrier tissues but also home to inflammatory sites. How this trafficking is regulated is unclear. Here the authors show that the dynamic expression of chemokine receptors CCR2 and CCR6 differentiates \u03b3\u03b4T17 cell trafficking patterns at homeostasis and in inflammatory scenarios. In models of autoimmunity, cancer and infection, activation-induced downregulation of CCR6 releases \u03b3\u03b4T17 cells from their homeostatic immunosurveillance trafficking circuit through the skin and circulation, which then enhances their CCR2-dependent homing to inflamed tissue. Therefore, the dynamic interplay between CCR6 and CCR2 expression defines \u03b3\u03b4T17 cell trafficking patterns between resting and activated states.\u2212CCR2+ phenotype marking the encephalitogenic granulocyte\u2013macrophage colony-stimulating factor/interferon-\u03b3-producing population221Il17aCre \u00d7 Rosa26eYFP mice, where Il17a expression drives permanent marking of cells with eYFP\u2212CD44hi phenotype, characteristic of \u03b3\u03b4T17 cells (+ and V\u03b36+ \u03b3\u03b4T17 cell subsets as distinguished by both V\u03b34 expression and CD3/T-cell receptor (TCR) expression level, as previously reported (\u2018CD3bright staining')ex vivo chemotaxis (\u2212CCR2+) were prominent in lung and gut-associated tissues (Ccr2-deficiency inhibited \u03b3\u03b4T17 cell recruitment to the CNS at both onset and peak disease (Ccr6 and Ccr2 (Ccr6\u2212/\u2212Ccr2\u2212/\u2212) did not further affect \u03b3\u03b4T17 cell infiltration in either model . CCR2-dres (LNs) . Similar disease , time po disease . CCR2 aper model .Ccr2\u2212/\u2212Ccr6\u2212/\u2212 mice exhibited enhanced tumour growth, while Ccr2\u2212/\u2212 and Ccr2\u2212/\u2212Ccr6\u2212/\u2212 mice had decreased EAE severity and Ccr2\u2212/\u2212 \u03b3\u03b4T17 cells were co-transferred into B16 melanoma-bearing recipients. While donor \u03b3\u03b4T17 cells recovered from spleen retained the input ratio, Ccr2\u2212/\u2212 \u03b3\u03b4T17 cells did not migrate efficiently to tumours. This observation was true for both V\u03b34+ and V\u03b36+ subsets than WT at 72\u2009h post-infection , a population previously reported to produce IL-17 and distinct from TCRhi dendritic epidermal T cellslo cells were entirely marked by eYFP in Il17aCre \u00d7 Rosa26eYFP mice, despite negligible IL-17A production following ex vivo restimulation (Ccr6-deficiency reduced the number of both V\u03b34+ and V\u03b34\u2212 (V\u03b36+) \u03b3\u03b4Tlo cells, although the ratio was skewed slightly towards V\u03b36+ cells was highly expressed in resting \u03b3\u03b4T17 cells but was downregulated by 24\u2009h of activation. Batf and Prdm1 (Blimp1) were rapidly upregulated by 24\u2009h, while Irf8 and Irf4 were upregulated by 48\u2009h, although Irf4 was already present in resting \u03b3\u03b4T17 cells. Expression of Eomes and Tbx21 (T-bet) at rest or following activation was minimal mice were bred at the University of Adelaide animal facility. Irf4\u2212/\u2212, Irf8\u2212/\u2212, Batf\u2212/\u2212 and LckCrePrdm1fl/fl mice were bred at the WEHI animal facility. Mice were age- and gender-matched and used at 6\u201314 weeks of age. Experiments were conducted with approval of the University of Adelaide Animal Ethics Committee.C57Bl/6 (WT) and Ly5.1 mice were purchased from Animal Resource Centre or bred at the University of Adelaide animal facility. 35\u201355 (GL Biochem) in phosphate-buffered saline (PBS) emulsified 1:1 in complete Freund's adjuvant, coupled with i.p. injection of 300\u2009ng Pertussis toxin (Sapphire Bioscience) on days 0 and 2. Mice were analysed at clinical scores of 0.5 (onset) and 2\u20133 (peak) in wild type (WT) mice, where scoring criteria were: 0.5 tremor, 1 partially limp tail, 2 fully limp tail, 2.25 unable to right, 2.5 sprawled hindlimbs, 2.75 one hindlimb paralysed, 3 both hindlimbs paralysed, 3.5 one forelimb paralysed. B16.F10 melanoma cells were cultured in RPMI 1640 containing 10% fetal calf serum (FCS) and 5 \u00d7 104 cells were injected subcutaneously into mice at four sites. S. pneumoniae EF3030 was grown to a D600 of 0.18 in nutrient broth with 10% horse serum at 37\u2009\u00b0C 5% CO2 and stored at \u221280\u2009\u00b0C. Stocks were defrosted and 1 \u00d7 106 colony-forming units were delivered intranasally. Bacterial load was determined by serial dilution of concentrated NW onto blood agar with 5\u2009\u03bcg\u2009ml\u22121 gentamicin (Sigma).Mice were immunized for chronic EAE by subcutaneous injection of 100\u2009\u03bcg of MOGg. The following digestions were performed at 37\u2009\u00b0C with 30U\u2009ml\u22121 DNase (Sigma). Epidermis and dermis from ears or shaved trunk skin were separated by incubation in 0.375% tryspin for 2\u2009h at 37\u2009\u00b0C, and then digested separately with 85\u2009\u03bcg\u2009ml\u22121 Liberase TM (Roche) for 1\u2009h. Tumours and perfused lungs were digested in 2\u2009mg\u2009ml\u22121 collagenase (Sigma) for 1\u2009h. NP tissue between the nose tip and eyes was dissected following removal of nasal-associated lymphoid tissue, digested with 2\u2009mg\u2009ml\u22121 collagenase for 1.5\u2009h and separated by a 40/80% Percoll gradient at 600g. Livers were pressed through 70\u2009\u03bcm filters and then lymphocytes were isolated by a 37.5% Percoll gradient at 850g. Flushed and longitudinally opened small intestine free of Peyer's patches was washed in PBS then incubated in 5\u2009mM EDTA for 40\u2009min at 37\u2009\u00b0C to remove epithelium, before remaining lamina propria was digested with 0.5\u2009mg\u2009ml\u22121 collagenase for 1.5\u2009h at 37\u2009\u00b0C and separated over a 40/80% Percoll gradient at 1,000g to isolate lamina propria lymphocytes. Peritoneal exudate cells were collected by 3 \u00d7 1\u2009ml PBS washes.Single-cell suspensions were prepared from lymphoid organs by pressing through 70\u2009\u03bcm filters. Peripheral blood was collected into heparinized Vacutainer tubes (BD). Red blood cells were lysed as required. CNS from PBS-perfused mice was pressed through 70\u2009\u03bcm filters and then separated over a 30/70% Percoll (GE) gradient at 5006 lymphocytes per well using antibodies and other reagents detailed in \u22121 PMA (Life Technologies), 1\u2009nM ionomycin (Life Technologies) and 1/1,500 GolgiStop (BD) for 4\u2009h at 37\u2009\u00b0C. Cells were washed in PBS and stained with Near Infrared fixable dye diluted 1/1,000 (Life Technologies) for 15\u2009min at room temperature. Cells were then washed with FACS buffer and blocked with mouse \u03b3-globulin (m\u03b3g) (200\u2009\u03bcg\u2009ml\u22121) for 5\u2009min at room temperature. All subsequent steps were incubated at 4\u2009\u00b0C. For purified antibodies, cells were stained with purified antibody for 20\u201360\u2009min, washed in FACS buffer, stained with secondary antibody in m\u03b3g (200\u2009\u03bcg\u2009ml\u22121) and normal mouse serum (NMS) (1%) for 20\u2009min, washed in FACS buffer and blocked with rat \u03b3-globulin for 15\u2009min. Cells were stained with directly conjugated and biotinylated antibodies for 20\u2009min. In the case of biotinylated antibodies, cells were then washed in FACS buffer and stained with streptavidin for 15\u2009min. Cells were then washed in PBS 0.04% azide. For intracellular cytokine staining, cells were incubated in Cytofix/Cytoperm (BD) for 20\u2009min, washed in Permwash (BD) and stained with intracellular directly conjugated antibodies for 20\u2009min. For transcription factor staining, cells were incubated in Foxp3 kit perm buffer (eBioscience) for 30\u2009min to overnight, and then washed in Foxp3 kit permwash (eBioscience). Cells were then stained with directly conjugated \u03b1-transcription factor antibodies in NMS (2%) and normal rat serum (2%), and then washed in PBS 0.04% azide. All stains were washed in PBS 0.04% azide, resuspended in PBS 1% paraformaldehyde and stored at 4\u2009\u00b0C in the dark.Single-cell suspensions were stained in 96-well round- or v-bottom plates (Corning) at 2 \u00d7 10Ccr2\u2212/\u2212 \u03b3\u03b4T17 cells. CCR2 and CCR6 gating was determined by relevant isotype controls. For measurement of fluorescence intensity, relevant isotype control geometric mean fluorescence intensity was subtracted from raw geometric mean fluorescence intensity value. For in vivo proliferation analysis, mice were given 2\u2009mg BrdU i.p. and then drinking water with 0.8\u2009mg\u2009ml\u22121 BrdU 2% glucose. Following restimulation and surface staining, cells were permeabilized, DNase-treated and stained with \u03b1-BrdU (BD) according to the manufacturer's instructions. For in vitro proliferation analysis, cells were labelled with Cell Proliferation Dye (eBioscience) according to the manufacturer's instructions. Flow cytometry was acquired on a BD LSR II or FACSAria and analysed with FlowJo (Treestar). Gating strategies are detailed in Specificity of \u03b1-CCR2 was confirmed by negative staining on 6 cells per ml in RPMI 1640 containing 10% FCS, antibiotics, 1 \u00d7 Glutamax (Gibco), 10\u2009mM HEPES (SA Pathology), 1\u2009mM sodium pyruvate, 54 pM \u03b2-mercaptoethanol and 1 \u00d7 non-essential amino acids (Gibco) with 5\u2009ng\u2009ml\u22121 recombinant (r)IL-23 (eBioscience), 5\u2009ng\u2009ml\u22121 rIL-1\u03b2 (Miltenyi Biotec) and 10\u2009\u03bcg\u2009ml\u22121 \u03b1-IFN-\u03b3 (BioXCell) in 96-well round-bottom plates coated with 1\u2009\u03bcg\u2009ml\u22121 \u03b1-TCR-\u03b3\u03b4 for 3 days. Cells were washed and re-seeded on fresh plastic at 1 \u00d7 106 cells per ml for a further 3 days as above without TCR-\u03b3\u03b4 stimulation. Cells were then washed and re-seeded in 20\u2009ng\u2009ml\u22121 rIL-7 (Peprotech) and 10\u2009\u03bcg\u2009ml\u22121 \u03b1-IFN-\u03b3 for a further 3 days. pMIG, pMIG-Rorc and pMIG-Ccr6 (cloned from mouse Ccr6 cDNA) were transfected into EcoPack 2 293 cells with Lipofectamine 2000 (ThermoFisher), and supernatant collected after 48\u2009h. \u03b3\u03b4T17 cells at days 4 and 5 of culture were centrifuged at 2,500\u2009r.p.m. (30\u2009\u00b0C for 1.5\u2009h) in supernatant with 8\u2009\u03bcg\u2009ml\u22121 polybrene (Sigma) in flat-bottom 96 well trays before being returned to culture.Pooled spleen and LN cells were cultured at 1 \u00d7 10Ccr2\u2212/\u2212 and Ccr6tg trafficking experiments, 1\u20132 \u00d7 106 each of in vitro-expanded F1 (CD45.1+CD45.2+) and Ccr2\u2212/\u2212(CD45.2+), or transduced control and Ccr6tg \u03b3\u03b4T17 cells (F1 or WT), were mixed and transferred i.v. into Ly5.1 (CD45.1+) recipient mice d5 post-challenge with B16 melanoma, d8\u201310 post EAE induction or 24\u2009h post-S. pneumoniae infection. \u03b3\u03b4T17 cell infiltration of target organs was analysed 24\u201348\u2009h post-transfer, and CD45 congenic ratios were normalized to input sample. For S. pneumoniae Tcrd\u2212/\u2212 reconstitution, in vitro-expanded WT and Ccr2\u2212/\u2212 \u03b3\u03b4T17 cells were further purified by MACS (Miltenyi Biotec) before 3 \u00d7 106 cells were transferred into separate Tcrd\u2212/\u2212 hosts 24\u2009h prior to infection. For na\u00efve dermis trafficking experiments, 5\u201310 \u00d7 107 fresh WT or Ccr6\u2212/\u2212 lymphocytes or 3 \u00d7 107 72\u2009h IL-23/IL-1\u03b2-stimulated WT lymphocytes were transferred into separate unimmunized Ly5.1 mice and analysed 36\u2009h later. Number of recovered cells was normalized to number of \u03b3\u03b4T17 cells transferred.For Tumour and NP supernatants from digested samples and supernatants from filtered CNS were supplemented with protease inhibitors (Sigma) and stored at \u221280\u2009\u00b0C. Mouse CCL2 Duoset ELISA (R&D) was conducted according to the manufacturer's instructions.Il17aCre \u00d7 Rose26eYFP mice were enriched by MACS using mouse TCR\u03b3\u03b4+ isolation kit , and then sorted using a BD FACSAria. Na\u00efve CD4+ T cells were sorted from WT splenocytes, and skin stromal populations were sorted from digested epidermal and dermal suspensions from WT mice. Sorting strategies are detailed in \u2212(CT target\u2013CT reference) where reference was Rplp0.\u03b3\u03b4 T cells from 6 splenocytes or 2 \u00d7 105in vitro-expanded \u03b3\u03b4T17 cells were loaded into the upper chambers and plates were incubated at 37\u2009\u00b0C for 3\u2009h. Lower wells were harvested and stained for flow cytometry. CountBrite beads (Invitrogen) were added to samples prior to acquisition to normalize event counts. Chemotaxis index was calculated as number of gated events divided by number in 0 chemokine control.Splenocytes were rested in complete RPMI for 3\u20134\u2009h at 37\u2009\u00b0C, washed and suspended in chemotaxis buffer . \u03b3\u03b4T17 cells from culture were washed and suspended in chemotaxis buffer. CCL2 or CCL20 (from the late Prof. Ian Clark-Lewis) were diluted in chemotaxis buffer and loaded into the lower chambers of 96-well 5\u2009\u03bcm pore transwell plates (Corning). 2 \u00d7 10L-glutamine, \u03b2-mercaptoethanol) at 2.5 \u00d7 106 cells per ml with 10\u2009ng\u2009ml\u22121 rIL-23 (eBioscience), 10\u2009ng\u2009ml\u22121 rIL-1\u03b2 (Miltenyi Biotec), 20\u2009ng\u2009ml\u22121 rIL-7 (Peprotech), 10\u2009ng\u2009ml\u22121 rIL-12 (R&D), 10\u2009ng\u2009ml\u22121 rIL-18 (R&D) and/or with pre-coating in 1\u2009\u03bcg\u2009ml\u22121 \u03b1-TCR-\u03b3\u03b4 (Biolegend) for up to 72\u2009h at 37\u2009\u00b0C. For mitomycin C pre-treatment, cells were first incubated with 10\u2009\u03bcg\u2009ml\u22121 mitomycin C (Sigma) in complete IMDM at 2 \u00d7 107 cells per ml for 2\u2009h at 37\u2009\u00b0C before extensive washing.Splenocytes were cultured in complete IMDM .Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.Supplementary Figures and Supplementary Tables"} +{"text": "In the crystal, N\u2014H\u22efO, O\u2014H\u22efO and N\u2014H\u22efF hydrogen bonds and weak \u03c0\u2013\u03c0 inter\u00adactions connect chains of mol\u00adecules into a three-dimensional network.The title compound {systematic name: 2-[\u00adoxy]acetic acid}, C 7H5Cl2FN2O3, the mean plane of the carb\u00adoxy\u00adlic acid substituent and the pyridyl ring plane subtend a dihedral angle of 77.5\u2005(1)\u00b0. In the crystal, pairs of O\u2014H\u22efO hydrogen bonds form inversion dimers with R22(8) ring motifs. These are extended into chains along [011] by N\u2014H\u22efF hydrogen bonds. In addition, inter\u00admolecular N\u2014H\u22efO hydrogen bonds and weak \u03c0\u2013\u03c0 inter\u00adactions [ring centroid separation = 3.4602\u2005(9)\u2005\u00c5] connect these chains into a three-dimensional network.In the title pyridine herbicide {systematic name: 2-[\u00adoxy]acetic acid}, C It is widely used on cereal crops, olive trees and fallow croplands to control broad-leaf weeds \u2005\u00c5; symmetry code: (iv) \u2212x, \u2212y\u00a0+\u00a02, \u2212z], resulting in a three-dimensional network structure = 0.84\u2005\u00c5, Uiso = 1.5Ueq(C) for the O\u2014H group, d(N\u2014H) = 0.88\u2005\u00c5, Uiso = 1.2Ueq(C) for the amine group, and d(C\u2014H) = 0.99\u2005\u00c5, Uiso = 1.2Ueq(C) for the CH2 group.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016018533/sj5515sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989016018533/sj5515Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016018533/sj5515Isup3.cmlSupporting information file. DOI: 1518035CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystals of both compounds, mol\u00adecules are linked 30H18N2O5S, (I), and C27H18N2O4S2, (II), are carbazole derivatives with a phenyl\u00adsulfonyl group and a nitro\u00adphenyl group attached to the carbazole moiety in identical positions in both mol\u00adecules. A benzo\u00adfuran ring system in (I) and a methyl\u00adthio\u00adphene ring in (II) are fused with the respective carbazole moieties on the same sides. The mean plane of the carbazole ring system makes a dihedral angle of 3.17\u2005(7)\u00b0 with the benzo\u00adfuran ring system in (I) and a dihedral angle of 3.39\u2005(11)\u00b0 with the methyl\u00adthio\u00adphene ring in (II), implying that both fused units are essentially planar. The mean planes of the carbazole ring systems in both the compounds are almost orthogonal to the respective nitro-substituted phenyl rings, making dihedral angles of 75.64\u2005(10) and 77.63\u2005(12)\u00b0 in compounds (I) and (II), respectively. In (I), the phenyl\u00adsulfonyl ring system is positionally disordered with a refined occupancy ratio of 0.63\u2005(2):0.37\u2005(2). In both compounds, the mol\u00adecular structures are stabilized by intra\u00admolecular C\u2014H\u22efO hydrogen bonds, generating S(6) ring motifs with the sulfone group O atoms. In the crystal of compound (I), mol\u00adecules are linked by pairs of C\u2014H\u22efO hydrogen bonds, which generate R22(18) inversion dimers, and inter\u00adconnected by C(14) chains running along the c-axis direction, whereas in compound (II), the C\u2014H\u22efO hydrogen bonds generate R43(37) ring motifs. In the crystals of both compounds, C\u2014H\u22efO hydrogen-bonded sheets are formed lying parallel to (10-1). In addition, C\u2014H\u22ef\u03c0 and offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.7158\u2005(14)\u2005\u00c5 in (I) and 3.9040\u2005(15)\u2005\u00c5 in (II)] are also present in the crystals of both compounds.The title compounds, C Carbazole derivatives exhibit various biological activities such as anti\u00adtumor 2 groups are inclined to the benzene ring (C19\u2013C24) to which they are attached by 9.8\u2005(4)\u00b0 in (I)The mean planes of the carbazole ring systems make dihedral angles of 3.17\u2005(7) and 3.39\u2005(11)\u00b0, respectively, with the benzo\u00adfuran ring in (I)sp2 bond lengths are longer than the mean value of 1.355\u2005(14)\u2005\u00c5 for N\u2014C bond lengths ring motifs with the sulfone oxygen atoms (Tables 1The sums of the bond angles around atom N1 are 349.58\u00b0 in (I) Tables 1 and 2 \u25b8.via pairs of C20\u2014H20\u22efO2ii hydrogen bonds chains. These inter\u00adactions result in the formation of sheets parallel to = 3.7158\u2005(14)\u2005\u00c5, inter\u00adplanar distance = 3.472\u2005(1)\u2005\u00c5, slippage = 1.324\u2005(11)\u2005\u00c5; Cg4 is the centroid of the C7\u2013C12 ring]; see Table\u00a01In the crystal of compound (I)s Table\u00a01, formingi and C14\u2014H14\u22efOii4 hydrogen bonds = 3.9040\u2005(15)\u2005\u00c5, inter\u00adplanar distance = 3.791\u2005(1)\u2005\u00c5, slippage 0.932\u2005\u00c5; Cg4 is the centroid of the C7\u2013C12, ring]; see Table\u00a02In the crystal of compound (II)s Table\u00a02, which rfs Fig.\u00a04. The cryrs Fig.\u00a05, which aet al., 2016H-[1] benzo\u00adfuran\u00adcarbazole and 47 hits for 9-(phenyl\u00adsulfon\u00adyl)-9H-carbazole. However, the compound 7-phenyl\u00adsulfonyl-7H-benzo\u00adfuran\u00adcarbazole H-thienocarbazole et al., 2011A search of Cambridge Structural Database : A solution of [3-(4-nitro\u00adbenzo\u00adyl)-1-(phenyl\u00adsulfon\u00adyl)-1H-indol-2-yl]methyl pivalate , anhydrous SnCl4 and benzo\u00adfuran in dry DCE (10\u2005ml) was stirred at room temperature under nitro\u00adgen for 3\u2005h. After completion of the reaction (monitored by TLC), it was poured into ice\u2013water (100\u2005ml). The organic layer was separated and the aqueous layer was extracted with DCM (2 \u00d7 20\u2005ml). The combined extract was washed with water (3 \u00d7 50ml) and dried (Na2SO4). Removal of the solvent by column chromatographic purification yielded compound (I)Compound (II): A solution of [3-(4-nitro\u00adbenzo\u00adyl)-1-(phenyl\u00adsulfon\u00adyl)-H-indol-2-yl]methyl pivalate , anhydrous SnCl4 and 2-methyl\u00adthio\u00adphene in dry DCE (10\u2005ml) was stirred at room temperature under nitro\u00adgen atmosphere for 3\u2005h. After the completion of the reaction (monitored by TLC), it was poured into ice\u2013water (100\u2005ml), the organic layer was separated and the aqueous layer was extracted with DCM (2 \u00d7 20ml). The combined extract was washed with water (3 \u00d7 50\u2005ml) and dried (Na2SO4). Removal of the solvent by column chromatographic purification yielded compound (II)Uiso(H)= 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. In compound (I)SHELXL97; Sheldrick, 2008Crystal data, data collection and structure refinement details for compounds (I)10.1107/S2056989016016996/su5333sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989016016996/su5333Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016016996/su5333IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989016016996/su5333Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989016016996/su5333IIsup5.cmlSupporting information file. DOI: 1473995CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II cation displays a distorted octa\u00adhedral geometry coordinated by four pyridine N atoms from four individual ligands and two O atoms from two monodentate nitrate anions. Two symmetry-related ligands are connected by two symmetry-related CoII cations, forming a 20-membered cyclic dimer. These cyclic dimers are connected to each other by sharing CoII atoms, forming a looped chain. In the crystal, adjacent looped chains are connected by inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions and C\u2014H\u22ef\u03c0 and C\u2014H\u22efO hydrogen bonds, resulting in the formation of a three-dimensional supra\u00admolecular architecture.The reaction of cobalt(II) nitrate with bis\u00ad(pyridin-3-ylmeth\u00adyl)sulfane ligand, afforded a one-dimensional looped polymeric chain. The Co 3)2(C12H12N2S)2]n, contains a bis\u00ad(pyridin-3-ylmeth\u00adyl)sulfane (L) ligand, an NO3\u2212 anion and half a CoII cation, which lies on an inversion centre. The CoII cation is six-coordinated, being bound to four pyridine N atoms from four symmetry-related L ligands. The remaining coordination sites are occupied by two O atoms from two symmetry-related nitrate anions in a monodentate manner. Thus, the CoII centre adopts a distorted octa\u00adhedral geometry. Two symmetry-related L ligands are connected by two symmetry-related CoII cations, forming a 20-membered cyclic dimer, in which the CoII atoms are separated by 10.2922\u2005(7)\u2005\u00c5. The cyclic dimers are connected to each other by sharing CoII atoms, giving rise to the formation of an infinite looped chain propagating along the [101] direction. Inter\u00admolecular C\u2014H\u22ef\u03c0 (H\u22efring centroid = 2.89\u2005\u00c5) inter\u00adactions between one pair of corresponding L ligands and C\u2014H\u22efO hydrogen bonds between the L ligands and the nitrate anions occur in the looped chain. In the crystal, adjacent looped chains are connected by inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid-to-centroid distance = 3.8859\u2005(14)\u2005\u00c5] and C\u2014H\u22ef\u03c0 hydrogen bonds (H\u22efring centroid = 2.65\u2005\u00c5), leading to the formation of layers parallel to (101). These layers are further connected through C\u2014H\u22efO hydrogen bonds between the layers, resulting in the formation of a three-dimensional supra\u00admolecular architecture.The asymmetric unit of the title compound, [Co(NO The CoII cation is coordinated by four pyridine N atoms from four symmetry-related L ligands. In addition, the CoII cation binds to two O atoms of two symmetry-related monodentate nitrate anions, forming a distorted octa\u00adhedral CoN4O2 coordination. Selected bond lengths and angles around the Co1 atom are listed in Table\u00a01II centre are tilted by 70.75\u2005(7)\u00b0 with respect to each other \u2005\u00c5. The cyclic dimers are connected by sharing CoII atoms, leading to the formation of an infinite looped chain propagating along the [101] direction. An inter\u00admolecular C7\u2014H7B\u22ef Cg2i inter\u00adactions between one pair of corresponding L ligands and several C\u2014H\u22efO hydrogen bonds between the L ligands and the NO3\u2212 anions (Table\u00a02Two symmetry-related s Table\u00a02 contribuCg1\u22efCg1ii = 3.8859\u2005(14)\u2005\u00c5; yellow dashed lines in Fig.\u00a02Cg1 is the centroid of atoms N1/C1\u2013C5; symmetry code: (ii) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01] together with inter\u00admolecular C6\u2014H6A\u22efCg2iii hydrogen bonds , generating layers parallel to (101). Neighboring layers are packed by C1\u2014H1\u22efO3iv hydrogen bonds between pyridine H atoms and nitro\u00adgen O atoms, resulting in the formation of a three-dimensional supra\u00admolecular architecture.Adjacent looped chains in the structure are connected by inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions between the N1-pyridine rings [L ligand was synthesized according to a literature method solution of the L ligand with Co(NO3)2\u00b76H2O in a 2:1 molar ratio.The sp2\u2014H and 0.97\u2005\u00c5 for methyl\u00adene C\u2014H with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017014980/hg5499sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989017014980/hg5499Isup2.hklStructure factors: contains datablock(s) I. DOI: 1580230CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound consists of an di\u00adethyl\u00adammonium\u2013aniline\u00adsulfonate ion pair and a zwitterionic aniliniumsulfonate mol\u00adecule. 4H12N+\u00b7C6H6NO3S\u2212\u00b7C6H7NO3S, consists of an ion pair and a zwitterionic neutral mol\u00adecule. The cation adopts an extended conformation [C\u2014C\u2014N\u2014C torsion angles = 177.1\u2005(3) and \u2212178.4\u2005(3)\u00b0]. In the crystal, the components are linked by N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, generating a three-dimensional network, which is consolidated by weak C\u2014H\u22efO inter\u00adactions.The title compound, C The bi-layers are then linked through N1\u2014H1NA\u22efO2, N1\u2014H1NB\u22efO5 and N1\u2014H1NB\u22efO6 hydrogen bonds, yielding a three-dimensional network inter\u00adactions consolidate the packing in the crystal . Examination of the packing reveals layers of diethyl ammonium cation sandwiched between bi-layers of aniline sulfate moieties. The key hydrogen bonds establishing the three-dimensional array are the contacts to sulfonate oxygen atoms and the N2\u22efN3 aniline inter\u00adactions. All amine hydrogen atoms form good hydrogen-bond contacts to neighboring hydrogen-bond acceptor atoms.The zwitterionic aniliniumsulfonate and the aniline\u00adsulfonate anion are connected through N2\u2014H2s Table\u00a01 giving srk Fig.\u00a02. Some weet al., 2016et al., 2008A search of the Cambridge Structural Database I. DOI: 10.1107/S2056989016018041/hb7622Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016018041/hb7622Isup3.cmlSupporting information file. DOI: 1515845CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-hy\u00addroxy\u00adquinoline-2-carboxamide mol\u00adecule has a nearly planar structure [maximum deviation = 0.062\u2005(1)\u2005\u00c5] and only the hy\u00addroxy H atom deviates from the mol\u00adecule plane.The 10H8N2O2\u00b7H2O, consists of an N-hy\u00addroxy\u00adquinoline-2-carboxamide mol\u00adecule in the keto tautomeric form and a water mol\u00adecule connected through an O\u2014H\u22efO hydrogen bond. The N-hy\u00addroxy\u00adquinoline-2-carboxamide mol\u00adecule has a nearly planar structure [maximum deviation = 0.062\u2005(1)\u2005\u00c5] and only the hy\u00addroxy H atom deviates significantly from the mol\u00adecule plane. In the crystal, \u03c0\u2013\u03c0 stacking between the aromatic rings [inter\u00adcentroid distance = 3.887\u2005(1)\u2005\u00c5] and inter\u00admolecular O\u2014H\u22efO hydrogen bonds organize the crystal components into columns extending along the b-axis direction.The title compound, C The carbonyl group possesses a Z conformation against the N1 atom of the quinoline moiety and E conformation against the hy\u00addroxy oxygen atom [torsion angles O2\u2014N2\u2014C10\u2014O1 = 0.8\u2005(2)\u00b0 and N1\u2014C9\u2014C10\u2014O1\u00a0\u2212177.33\u2005(14)\u00b0]. The N-hy\u00addroxy\u00adquinoline-2-carboxamide mol\u00adecule has an almost planar structure (non-hydrogen atoms are planar to within 0.03\u2005\u00c5). Only the H atom of the OH group deviates significantly from the mol\u00adecular plane: the C\u2014N\u2014O\u2014H torsion angle of \u221275.1\u2005(13)\u00b0 is defined by the O\u2014H\u22efO hydrogen bond between hy\u00addroxy group and the water mol\u00adecule. The C\u2014N and C\u2014C bond lengths in the quinoline moiety are typical for 2-substituted pyridine derivatives \u2005\u00c5, inter\u00adcentroid distance 3.887\u2005(1)\u2005\u00c5, displacement 1.846\u2005(1)\u2005\u00c5]. These columns are linked pairwise by the O\u2014H\u22efO hydrogen bonds global, I. DOI: 10.1107/S2056989017005618/xu5902Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017005618/xu5902Isup3.cmlSupporting information file. DOI: 1543825CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The persence of C\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adactions in the crystals lead to dimeric aggregates in (I) and supra\u00admolecular chains (II).The metal coordination geometry in each of the title mol\u00adecules, [Sn(C viz. [Sn(C6H5)3(C8H16NS2)], (I), and [Sn(C6H5)3(C10H12NS2)], (II), are described. The di\u00adthio\u00adcarbamate ligand in each mol\u00adecule coordinates in an asymmetric fashion resulting in heavily distorted tetra\u00adhedral C3S coordin\u00adation geometries for the Sn atoms, with the distortions traced to the close approach of the non-coordinating thione-S atom. The mol\u00adecular packing in both compounds features C\u2014H\u22ef\u03c0(Sn-phen\u00adyl) inter\u00adactions. In (I), the donors are Sn-phenyl-C\u2014H groups leading to centrosymmetric aggregates, while in (II), the donors are both Sn-phenyl-C\u2014H and methyl-C\u2014H groups leading to supra\u00admolecular chains propagating along the b axis. The identified aggregates assemble into their respective crystals with no directional inter\u00adactions between them. An analysis of the Hirshfeld surfaces show distinctive patterns, but an overwhelming predominance (>99% in each case) of H\u22efH, C\u22efH/H\u22efC and S\u22efH/H\u22efS contacts on the respective Hirshfeld surface.The crystal and mol\u00adecular structures of two tri\u00adphenyl\u00adtin di\u00adthio\u00adcarbamate compounds, A key inter\u00adest in di\u00adthio\u00adcarbamate compounds of both transition metals and main-group elements relates to their biological activity 3Sn[S2CN(Me)Hex] (I)6H5)3Sn[S2CN(CH3)CH2CH2Ph] (II)A vast array of different di\u00adthio\u00adcarbamate anions, a, is coordinated by three ipso-carbon atoms along with a di\u00adthio\u00adcarbamate ligand. As seen from Table\u00a01long \u2013 Sn\u2014Sshort) being 0.64\u2005\u00c5. This asymmetry is confirmed in the differences in the C\u2014S bond lengths with the C1\u2014S1 bond associated with the short Sn\u2014S1 contact, at 1.761\u2005(4)\u2005\u00c5, being significantly longer than the C1\u2014S2 bond, i.e. 1.688\u2005(4)\u2005\u00c5, involving the weakly bound S2 atom. If the S2 atom is ignored, the coordination geometry about the tin atom is distorted C3S tetra\u00adhedral with the range of angles being 90.00\u2005(11)\u00b0, for S1\u2014Sn\u2014C31, to 121.53\u2005(10)\u00b0, for S1\u2014Sn\u2014C11. The wide angle clearly reflects the influence of the close approach of the S2 atom, Fig.\u00a01a and Table\u00a013S2 donor set is almost perfectly inter\u00admediate between ideal square-pyramidal (SP) and trigonal\u2013bipyramidal (TP). This is qu\u00adanti\u00adfied in the value of \u03c4 = 0.52, which compares with the ideal values for SP and TP geometries of \u03c4 = 0.0 and 1.0, respectively , 178.5\u2005(4) and 178.9\u2005(5)\u00b0, respectively, indicate + anti-periplanar descriptors but, that of C5\u2014C6\u2014C7\u2014C8, i.e. \u221266.4\u2005(8)\u00b0, indicative of a \u2212 syn-clinal disposition.The tin atom in (I)b, resembles closely that described for (I)3S2 description pertains, the value of \u03c4 = 0.60 indicates a distortion towards TP. The phenyl\u00adethyl chain is kinked as seen in the N1\u2014C3\u2014C4\u2014C5 and C3\u2014C4\u2014C5\u2014C6 torsion angles of \u2212175.8\u2005(3) and 91.9\u2005(5)\u00b0, respectively.The mol\u00adecular structure of (II)a. Such cooperative C\u2014H\u22ef\u03c0(phen\u00adyl) embraces have been described for many phenyl-rich systems and in instances where six phenyl rings of two residues associate by edge-to-face inter\u00adactions, i.e. a six-fold embrace, the energies of stabilization can resemble or even exceed that provided by strong conventional hydrogen bonding PLATON et al., 2017i.e. n-hexyl in the former and phenyl\u00adethyl in the latter. These differences are readily discerned from the differently shaped Hirshfeld surfaces mapped over dnorm for (I)The Hirshfeld surface calculations for the tri\u00adphenyl\u00adtin di\u00adthio\u00adcarbamate derivatives (I)a reflect the presence of a weak C\u2014H\u22ef\u03c0 inter\u00adaction, as summarized in Table\u00a04C and n-hexyl atoms C7 and H7B are indicative of the short inter\u00adatomic H\u22efH and C\u22efH/H\u22efC contacts involving these atoms, as listed in Table\u00a04cf. (I)dnorm in Fig.\u00a05B and phenyl-C11 together with the pair of faint-red spots near the methyl-H2B and phenyl-C16 atoms in Fig.\u00a05a. The influence of other short inter\u00adatomic C\u22efH/H\u22efC contacts summarized in Table\u00a04a,b. The involvement of different atoms in the inter\u00admolecular inter\u00adactions in the crystals of (I)The faint-red spots near the phenyl-C33 and H26 atoms in Fig.\u00a04a, also highlight the different mol\u00adecular environments for the two mol\u00adecules. The significant contributions from H\u22efH, C\u22efH/H\u22efC and S\u22efH/H\u22efS contacts to the Hirshfeld surfaces of both (I)C and n-hexyl-H7B atoms in (I)de + di \u223c2.0\u2005\u00c5 in the delineated plot A and phenyl-H7, H9 and H23 atoms in comparatively weaker short inter\u00adatomic H\u22efH contacts, Table\u00a04de + di \u223c2.6\u2005\u00c5 in the fingerprint plot delineated into C\u22efH/H\u22efC contacts for (I)c, is the result of a short inter\u00adatomic contact between phenyl-C32 and -H23 atoms while the points corresponding to other short inter\u00adatomic contacts are merged within the plot. The presence of a pair of twin forceps-like tips at de\u00a0+\u00a0di \u223c\u00a02.7\u2005\u00c5 in the C\u22efH/H\u22efC delineated plot for (II)c, also indicates the involvement of methyl-H2A and -H2B, and phenyl-C7, -C8, -C11, -C16 and -C35 atoms in short inter\u00adatomic contacts, Table\u00a04d, that the pair of spikes at de\u00a0+\u00a0di \u223c 3.0\u2005\u00c5 for (I)de\u00a0+\u00a0di > 3.1\u2005\u00c5 for (II)The distinct distribution of points in the overall two-dimensional fingerprint plots for (I)et al., 2016N-hexyl-N-methyl\u00addithio\u00adcarbamate ligand reported in (I)i.e. dtcI, has been reported in the crystal structures of Ph2Sn(dtcI)2 compound, i.e. Hg(dtcII)3 in the Cambridge Structural Database short bond length is 2.47\u2005\u00c5 and the average Sn\u2014Slong bond length is 3.04\u2005\u00c5. This gives rise to an average \u0394(Sn\u2014S) of 0.57\u2005\u00c5. These values indicate the structures of (I)long are generally longer than usually observed. An analysis of the available crystallographic data showed the shortest Sn\u2014S1 bond length occurred in the structure of Ph3Sn(S2CNEt2) SnPh3 2} ; 26.39 [N\u2014(CH2)3CH2]; 22.6 [N\u2014CH2)4CH2]; 14.06 (hex\u00adyl\u2014CH3). 119Sn NMR (CDCl3): \u2212187.56.Synthesis of tri\u00adphenyl\u00adtin(IV)N-methyl-N-phenylethyl\u00addithio\u00adcarbamate (II) compound (II)N-methyl-N-phenylethyl\u00adamine in place of N-hexyl-N-methyl\u00adamine. Recrystallization was achieved by dissolving the compound in a chloro\u00adform/ethanol mixture (1:2 v/v). Yield: 67%, m.p. 387.5\u2013388.3\u2005K. Elemental analysis: calculated (%): C 60.0, H 4.9, N 2.5, S 11.4. Found (%): C 57.9, H 5.3, N 2.8, S 11.2. IR (KBr cm\u22121): 1452 \u03bd(C\u2014N), 977 \u03bd(C\u2014S), 502 \u03bd(Sn\u2014C), 488 \u03bd(Sn\u2014S). 1H NMR (CDCl3): \u03b4 7.43\u20137.77 ; 7.24\u20137.35 ; 4.06 ; 3.36 ; 3.09 . 13C NMR (CDCl3): \u03b4 196.61 (NCS2); 126.8\u2013142.3 (C-aromatic); 60.25 (NCH2); 44.59 (NCH2CH2); 33.12 (N\u2014CH3). 119Sn NMR (CDCl3) = \u2212183.84.Uiso(H) set to 1.2\u20131.5Ueq(C). For (I)\u22123, respectively, are located 0.95 and 0.86\u2005\u00c5 from the Sn atom. For (II)\u22123, respectively, are located 0.96 and 0.68\u2005\u00c5 from the C11 and Sn atoms, respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S2056989018005133/hb7745sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989018005133/hb7745Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989018005133/hb7745IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1833664, 1833663CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The cation and anion of the title salt are linked by an O\u2014H\u22efN hydrogen bond and a C\u2014H\u22efO inter\u00adaction, resulting in a high viscosity and a crystallization temperature slightly lower than ambient temperature. 3H8NO+\u00b7C2F6NO4S2\u2212, has ortho\u00adrhom\u00adbic (P212121) symmetry; the amino H atom of bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfon\u00adyl)amine (HNTf2) was transferred to the basic O atom of di\u00admethyl\u00adformamide (DMF) when the ionic liquid components were mixed. The structure displays an O\u2014H\u22efN hydrogen bond, which links the cation to the anion, which is reinforced by a non-conventional C\u2014H\u22efO inter\u00adaction, generating an R22(7) loop. A further very weak C\u2014H\u22efO inter\u00adaction generates an [001] chain.At 100\u2005K, the title mol\u00adecular salt, C The N2\u2014C4\u2014O5 angle does not deviate from the expected 120\u00b0 of an sp2-hybridized carbon atom [120.37\u2005(11)\u00b0]. The bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfon\u00adyl)amide anion features S1\u2014N1 and S2\u2014N1 bond distances of 1.6035\u2005(11) and 1.5947\u2005(11)\u2005\u00c5, respectively.The asymmetric unit consists of one bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfon\u00adyl)amide anion and one di\u00admethyl\u00adformamidium cation Fig.\u00a01: when thThe ion pair features two hydrogen bonds Table\u00a01. One is A CSD search found no structures that have the same ion pairing. Some structures feature the same bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfon\u00adyl)amide anion but different cations, which are usually metal complexes.2 (I)et al., 2014A literature procedure was followed to synthesize [(DMF)H]NTfUiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. The methyl groups were refined as rotating groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016012251/hb7586sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016012251/hb7586Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016012251/hb7586Isup3.cmlSupporting information file. DOI: 1496417CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, dications, anions and solvent water mol\u00adecules are connected via N/C/O\u2014H\u22efCl and N\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional network.In the title salt, C N,N-bis\u00ad(pyridin-4-ylmeth\u00adyl)cyclo\u00adhexane-1,4-di\u00adamine with hydro\u00adchloric acid in ethanol led to the formation of the title salt, C18H26N42+\u00b72Cl\u2212\u00b72H2O, which lies about a crystallographic inversion center at the center of the cyclo\u00adhexyl ring. The asymmetric unit therefore comprises one half of the N,N-bis\u00ad(pyridin-4-ylmeth\u00adyl)cyclo\u00adhexane-1,4-di\u00adammonium dication, a chloride anion, and a solvent water mol\u00adecule. In the dication, the two trans-(4-pyridine)\u2013CH2\u2013NH2\u2013 moieties occupy equatorial sites at the 1- and 4-positions of the central cyclo\u00adhexyl ring, which is in a chair conformation. The terminal pyridine ring is tilted by 27.98\u2005(5)\u00b0 with respect to the mean plane of the central cyclo\u00adhexyl moiety (r.m.s. deviation = 0.2379\u2005\u00c5). In the crystal, dications, anions, and solvent water mol\u00adecules are connected via N/C/O\u2014H\u22efCl and N\u2014H\u22efO hydrogen bonds together with C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional network.Treatment of The terminal pyridine ring is tilted by 27.98\u2005(5)\u00b0 with respect to the mean plane through the central cyclo\u00adhexyl moiety (r.m.s. deviation = 0.2379\u2005\u00c5). The distance between the two terminal pyridine nitro\u00adgen atoms in the dication is 15.864\u2005(2)\u2005\u00c5. This is slightly shorter than the N\u22efN separation [15.970\u2005(3)\u2005\u00c5] in the dication ligand of a one-dimensional zigzag-like CoII coordination polymer built up from alternate CoII ions and the dication of the title salt cyclo\u00adhexane-1,4-di\u00adamine, synthesized according to a literature method = 0.95\u2005\u00c5 for Csp2\u2014H, 0.99\u2005\u00c5 for methyl\u00adene, 1.00\u2005\u00c5 for methine H atoms, and were refined as riding with Uiso(H) = 1.2Ueq(C). The N- and O-bound H atoms involved in hydrogen bonding were located in difference Fourier maps and refined freely .Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016014626/sj5507sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989016014626/sj5507Isup2.hklStructure factors: contains datablock(s) I. DOI: 1504428CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Both mol\u00adecules adopt the ladder structure typical for this class of dimeric tetra\u00adorganodistannoxane di\u00adcarboxyl\u00adates. A nearly linear very short C\u2014H\u22efO contact gives rise to a chain-like arrangement of mol\u00adecules in the [111] directionThe asymmetric unit of [{Sn(C 4H9)2(C6H5COO)}2O]2, consists of two half molecules, completed by application of inversion symmetry. Both mol\u00adecules adopt a ladder structure typical for this class of dimeric tetra\u00adorganodistannoxane di\u00adcarboxyl\u00adates characterized by a centrosymmetric four-membered (Sn\u2014O)2 ring of rhomboidal shape that is extended on both sides by folded six-membered Sn\u2014O\u2014C rings. To a first approximation, both kinds of Sn atoms (Sni and Sno) are trigonal\u2013bipyramidally coordinated. The bond angles between the n-butyl groups are widened [135.64\u2005(7)\u2013146.20\u2005(7)\u00b0] in comparison with an ideal trigonal bipyramid. Sn\u2014O bond lengths within the {R2SnO3} coordination sphere depend strongly on the position of the corresponding O atom \u2013 axial (ax) or equatorial (eq) \u2013 as well as on the functionality of the carboxyl\u00adate groups which exhibit \u03bc2 (\u2013COOi) and \u03bc1 (\u2013COOo) coordination modes, respectively. In summary, the following sequence of distances [mean values] is found: d(Sno\u2014O\u03bc3)eq [2.024\u2005(2)\u2005\u00c5] < d(Sni\u2014O\u03bc3)eq [2.044\u2005(2)\u2005\u00c5] < d(Sni\u2014O\u03bc3)ax [2.158\u2005(6)\u2005\u00c5] < d(Sno\u2014O\u03bc1-carb)ax [2.182\u2005(6)\u2005\u00c5] < d(Sni\u2014O\u03bc2-carb)ax [2.250\u2005(2)\u2005\u00c5] \u2243 d(Sno\u2014O2-carb\u03bc)ax [2.247\u2005(12)\u2005\u00c5]. The n-butyl groups adopt an anti\u2013anti conformation with exception of two disordered outer n-butyl groups of the second mol\u00adecule which exhibit gauche\u2013anti and anti\u2013gauche conformations. Weak intra\u00admolecular Sn\u22efO inter\u00adactions between the different O atoms of the outer carboxyl groups with the inner, as well as outer, Sn atoms give rise to a strongly distorted octa\u00adhedral coordination at these Sn atoms. Inter\u00admolecular inter\u00adactions between the individual mol\u00adecules are restricted to van der Waals and O\u22efH\u2014C inter\u00adactions of which a nearly linear very short C\u2014H\u22efO contact between the H atom of the phenyl group of one of the mol\u00adecules with the outer non-coordinating C=O group of the other molecule is the most prominent. It gives rise to a chain-like arrangement of the mol\u00adecules along [111]. The two n-butyl groups attached to the outer Sn atom of one mol\u00adecule are disordered over two sets of sites with occupancies of 0.806\u2005(3)/0.194\u2005(3) and 0.702\u2005(3)/0.298\u2005(3).The asymmetric unit of the title compound, [{Sn(C The disorder of the two n-butyl groups was managed by a split model with site occupancies of 0.806\u2005(3)/0.194\u2005(3) and 0.702\u2005(3)/0.298\u2005(3). No further consideration will be made for the structural parameters of those n-butyl groups. It is noteworthy, however, that this disorder is caused by the conformational flexibility of the n-butyl group which adopts \u2013 in the case of the major/minor components \u2013 a gauche\u2013anti/anti\u2013gauche and anti-gauche/anti\u2013gauche conformation with respect to the Sn\u2014C\u03b1\u2014C\u03b2\u2014C\u03b3 and C\u03b1\u2014C\u03b2\u2014C\u03b3\u2014C\u03b4 torsion angles . This conformation of the disordered n-butyl groups is in contrast to the conformation of all other n-butyl groups of both dimers, which show exclusively an anti\u2013anti conformation . Structural parameters mean = 1.521\u2005(6)\u2005\u00c5, \u2329(C\u2014C\u03b2/\u03b3\u2014C)mean = 112.6\u2005(11)\u00b0 while bond angles at C\u03b1 range from 112.5\u2005(1) to 123.1\u2005(1)\u00b0. Sn\u2014C distances are in the range of 2.127\u2005(2)\u20132.134\u2005(2)\u2005\u00c5, mean value 2.130\u2005(3)\u2005\u00c5.The title compound Fig.\u00a01 crystallry Fig.\u00a01. With thes Fig.\u00a02b,c. Thion Fig.\u00a02a. Strucs Table\u00a01 within tladder structure typical for this class of tetra\u00adorganodistannoxane di\u00adcarboxyl\u00adates and two outer (Sno) tin atoms are \u2013 to a first approximation \u2013 fivefold, trigonal\u2013bipyramidally coordinated and linked together via two \u03bc2-coordinating oxygen atoms (Oi) and two chelating (\u2013COOi) carboxyl\u00adate groups. The structure is completed by two monodentate carboxyl\u00adate groups (\u2013COOo) attached to the outer tin atoms /75.78\u2005(5)\u00b0] angles at tin and obtuse ones [104.10\u2005(5)/104.22\u2005(5)\u00b0] at oxygen. Its rhomboidal shape with different Sn\u2014O distances results from the position of the \u03bc3-oxygen atom (O1/O3) within the trigonal\u2013bipyramidal coordination sphere of the inner tin atoms (Sn1/Sn3): bonds where the O atom is in an equatorial (eq) position are significantly shorter [2.042\u2005(1)/2.046\u2005(1)\u2005\u00c5] than those where the O atom is in an axial (ax) position [2.164\u2005(1)/2.152\u2005(1)\u2005\u00c5]. The second axially positioned Sn\u2014O bond at the inner tin atoms is even longer [2.251\u2005(1)/2.248\u2005(1)\u2005\u00c5] as it results from a coordinative bond of the oxygen atom (O11/O31) of the \u03bc2-benzoate ligand (\u2013COOi). In contrast to the prediction of the VSEPR concept, the bond angle between both equatorially positioned n-butyl groups is widened to 146.20\u2005(7)/141.73\u2005(7)\u00b0.The central, planar and centrosymmetric four-membered (Sn\u2013O)d(Sno\u2014O\u03bc3)eq [2.027\u2005(1)/2.022\u2005(1)\u2005\u00c5] < d(Sno\u2014O\u03bc1-carb)ax [2.175\u2005(1)/2.188\u2005(1)\u2005\u00c5] < d(Sno\u2014O\u03bc2-carb)ax [2.260\u2005(1)/2.235\u2005(1)\u2005\u00c5]. Bond angles between the n-butyl groups at the tin atoms are 142.49\u2005(7) and 135.64\u2005(7)\u00b0.The conformation of the outer, six-membered Sn\u2013O\u2013C rings is defined by an angle of 19.9\u2005(1)/23.4\u2005(1)\u00b0 between the O\u2013C\u2013O plane and the Sn\u2013O\u2013Sn plane Fig.\u00a04. In caseo), the C\u2014O bonds are of different strengths: the short, strong one [1.232\u2005(2)/1.222\u2005(3)\u2005\u00c5] indicates a localized C=O double bond whereas the long, weak one [1.307\u2005(2)/1.306\u2005(3)\u2005\u00c5] of the Sn-coordinating oxygen atom indicates a localized C\u2014O single bond. In the case of the bridging benzoate groups (\u2013COOi) both C\u2014O bonds are of almost equal lengths , in accordance with a delocalized \u03c0-system. In the two benzoate ligands, the carboxyl\u00adate groups and the phenyl groups are not co-planar, but are inclined to each other at angles of 15.1\u2005(2), 14.8\u2005(3)/3.9\u2005(3) and 17.3\u2005(1)\u00b0.The different coordination modes of both benzoate ligands are reflected in different C\u2014O bond lengths: in the case of the monodentate carboxyl\u00adate group (\u2013COOd(Sn\u22efO) = 2.7857\u2005(2)/2.7141\u2005(2)\u2005\u00c5] of the \u03bc1-O atoms (O21/O41) of the outer carboxyl\u00adate groups with the inner tin atoms (Sn1/Sn3), while those of the \u03bc0-O atoms (O22/O42) of the outer carboxyl\u00adate groups with the outer tin atoms (Sn2/Sn4) are still longer [2.8901\u2005(2)/2.9883\u2005(2)\u2005\u00c5]. Taking these weak inter\u00adactions into account, both kinds of Sn atoms adopt a strongly distorted octa\u00adhedral coordination. All bonding features (except the last ones) of the two mol\u00adecules are summarized in Fig.\u00a05n-butyl groups relative to the Sn\u2013O framework.Another characteristic feature of the mol\u00adecular structure comprises some additional, very weak inter\u00adactions [d(H\u22efO) = 2.487\u2005\u00c5], nearly linear [\u2329(C\u2014H\u22efO) = 172.6\u00b0] contact between the hydrogen atom H26 of a phenyl ring of mol\u00adecule 1 and the non-coordinating oxygen atom O42 of the second mol\u00adecule attracts attention as it leads to a chain-like arrangement of the two mol\u00adecules along [111] ]2O, have been extensively structurally characterized. The Cambridge Structural Database while for the di\u00adcarboxyl\u00adates benzoic acid derivatives (90) are the most studied. Even for the combination of R = nBu and R\u2032 = benzoic acid derivatives not less than 67 structures are described, but from the parent compound with R\u2032 = PhCOO\u2212, only the structure of the methyl compound (R = Me) has been completely characterized }2O]2 was obtained from an equimolar mixture of 0.300\u2005g (1.2\u2005mmol) of n-di\u00adbutyl\u00adtin oxide with 0.147\u2005g (1.2\u2005mmol) of benzoic acid in ethanol under reflux for 3.5\u2005h. After removal of the solvent, single crystals were obtained by recrystallization of the solid from ethanol/n-hexane. and displacement parameters were taken from the chemically equivalent C atoms of the major occupancy component.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017001505/zl2692sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017001505/zl2692Isup2.hklStructure factors: contains datablock(s) I. DOI: 1530058CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the former, the 4-methyl\u00adbenzene ring substituent makes a dihedral angle of 71.40\u2005(9)\u00b0 with the mean plane of the naphthalene ring system, while the phenyl ring substituent in the latter makes a dihedral angle of 67.08\u2005(12)\u00b0 with the mean plane of the anthracene ring system.The title compounds, 6-, and C26H16S, (II), are benzo\u00adthio\u00adphene derivatives in which the benzo\u00adthio\u00adphene moiety is fused with a naphthalene ring system in (I), and with an anthracene ring system in (II). In (I), the mean plane of the benzo\u00adthio\u00adphene ring system makes a dihedral angle of 2.28\u2005(6)\u00b0 with the naphthalene ring system, and a dihedral angle of 1.28\u2005(6)\u00b0 with the anthracene ring system in (II), showing that the fused units are essentially planar. In (I), the 4-methyl\u00adbenzene ring substituent makes a dihedral angle of 71.40\u2005(9)\u00b0 with the naphthalene ring system, while the phenyl ring substituent in (II) makes a dihedral angle of 67.08\u2005(12)\u00b0 with the anthracene ring system. In the crystals of both compounds, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, leading to the formation of slabs parallel to (001) in (I) and to zigzag chains along [001] in (II). There are also offset \u03c0\u2013\u03c0 inter\u00adactions present within the slabs in (I). In the crystal of (II), they link the chains, forming sheets parallel to (010). The triclinic polymorph of compound (II) has been reported .The title compounds, Cal., 2012. J. Org. In the crystal of (II)Cg2\u22efCg4iii = 3.711\u2005(2)\u2005\u00c5, inter\u00adplanar distance = 3.479\u2005(1)\u2005\u00c5, slippage = 1.21\u2005\u00c5; Cg3\u22efCg4iii = 3.741\u2005(2)\u2005\u00c5, inter\u00adplanar distance = 3.443\u2005(1)\u2005\u00c5, slippage = 1.22\u2005\u00c5; Cg2, Cg3 and Cg4 are the centroids of rings C1\u2013C3/C10\u2013C12, C1/C12\u2013C16 and C4\u2013C9, respectively; symmetry code: (iii) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01].In the crystals of both compounds, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions benzo[b]naphtho\u00adthio\u00adphene b]thio\u00adphen-3-yl){2-[pivalo\u00adyloxy(p-tol\u00adyl) meth\u00adyl]phen\u00adyl}methyl pivalate as a viscous liquid. Dipivalate (benzo[b]thio\u00adphen-3-yl){2-[pivalo\u00adyloxy(p-to\u00adyl) meth\u00adyl]phen\u00adyl}methyl pivalate upon inter\u00adaction with ZnBr2 followed by removal of solvent and column chromatographic purification gave 6-(p-tol\u00adyl)benzo[b]naphtho\u00adthiophene as a pale-green solid . Single crystals suitable for X-ray diffraction were prepared by slow evaporation of a solution of (I)The reduction of the diketone (benzo\u00adthio\u00adphen-3-yl)[2-(4-methyl\u00adbenzo\u00adyl)phen\u00adyl]methanone using sodium borohydride followed by work-up gave the diol. Dipivaloylation of the diol using pivaloyl chloride and tri\u00adethyl\u00adamine in the presence of a catalytic amount of DMAP (10\u2005mg) in dry DCM (20\u2005ml) led to the isolation of dipivalate (benzothio\u00adphen-2-yl)methanone using sodium borohydride followed by work-up gave the diol. Dipivaloylation of the diol using pivaloyl chloride and tri\u00adethyl\u00adamine in the presence of a catalytic amount of DMAP (10\u2005mg) in dry DCM (20\u2005ml) led to the isolation of dipivalate {2-[phen\u00adylmeth\u00adyl]phen\u00adyl}methyl pivalate as a thick liquid. Dipivalate {2-[phen\u00adylmeth\u00adyl]phen\u00adyl}methyl pivalate upon inter\u00adaction with ZnBr2 followed by removal of solvent and column chromatographic purification gave a new ortho\u00adrhom\u00adbic polymorph of 7-phenyl\u00adanthrabenzo[d]thio\u00adphene as a yellow solid. Single crystals suitable for X-ray diffraction were prepared by slow evaporation of a solution of the compound (II)The reduction of the diketone (2-benzoyl\u00adphen\u00adyl)(dibenzo[Uiso(H) = 1.5Ueq(methyl C) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details for compounds (I)10.1107/S2056989016012937/lh5819sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989016012937/lh5819Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989016012937/lh5819IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989016012937/lh5819Isup4.cmlSupporting information file. DOI: 1498519, 1498518CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The three mol\u00adecules have slightly varying overall conformations, all having trans conformations with respect to the C=N bond. In the crystal, the packing features N\u2014H\u22efN hydrogen bonds, which connect mol\u00adecules into chains extending along the c-axis direction, inter\u00adlinked by C\u2014H\u22ef\u03c0 inter\u00adactions (minimum H\u22efCg = 2.65\u2005\u00c5) into sheets lying parallel to (001).The title compound, C These are close to the literature value of 1.279\u2005\u00c5 for Csp2=Nsp2 bonds , 1.472\u2005(3) and 1.476\u2005(3)\u2005\u00c5, respectively. The comparative N1\u2014C16 bonds connecting the central benzene ring to the central C=N bond in A, B and C are 1.422\u2005(2), 1.419\u2005(2) and 1.420\u2005(2)\u2005\u00c5, respectively. The C14\u2014C15\u2014N1\u2014C16, torsion angles for the \u2013C\u2014C\u2014N\u2014C\u2013 bridge groups are \u2212178.47\u2005(17)\u00b0 (for A), \u2212176.35\u2005(17)\u00b0 (for B) and 178.31\u2005(17)\u00b0 (for C). The comparative dihedral angles between the anthracene ring system of the mol\u00adecule (defined by C1\u2013C14) and the benzene and phenyl rings (defined by C16\u2013C21 and C22\u2013C27) and between the benzene and phenyl rings, respectively, are 82.68\u2005(4), 73.76\u2005(5) and 25.63\u2005(11)\u00b0 in A, 80.10\u2005(4), 78.82\u2005(5) and 22.56\u2005(11)\u00b0 (in B) and 85.02\u2005(5), 81.66\u2005(5) and 16.25\u2005(11)\u00b0 (in C).The title compound, crystallizes with three independent mol\u00adecules = 2.65\u2005\u00c5; C\u2014H\u22efCg = 154\u00b0], forming layers lying parallel to (001) Fig.\u00a03.et al., 2016N-phenyl-p-phenyl\u00adenedi\u00adamine. Of these three compounds, N1-phenyl-N-4-(quinolin-2-yl\u00admethyl\u00adene)benzene-1,4-di\u00adamine {synonym: N-phenyl-4-[(quinolin-2-yl\u00admethyl\u00adene)amino]-aniline; WOJJIQ -(pyren-1-yl)-methyl\u00adidene]benzene-1,4-di\u00adamine of 9-anthracenecarboxaldehyde in 5\u2005ml of absolute ethanol was added dropwise under stirring. The mixture was stirred for 10\u2005min, two drops of glacial acetic acid were then added and the mixture was further refluxed for 2h. The resulting yellow precipitate was recovered by filtration, washed several times with small portions of ice-cold ethanol and then with diethyl ether to give 140\u2005mg (87%) of the title compound. Dark-yellow block-like crystals suitable for X-ray analysis were obtained within 3 days by slow evaporation of a solution in MeOH.80\u2005mg (0.435\u2005mmol) of Uiso(H)= 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016020612/zs2373sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016020612/zs2373Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989016020612/zs2373Isup3.cmlSupporting information file. DOI: 1524849CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The evolution of T1\u03c1 and of other endogenous contrast methods in the first month after reperfused myocardial infarction (MI) is uncertain. We conducted a study of reperfused MI in pigs to serially monitor T1\u03c1, T2 and T1 relaxation, scar size and transmurality at 1 and 4\u00a0weeks post-MI.n\u2009=\u200910) and 4\u00a0weeks (n\u2009=\u20095). Semi-automatic FWHM thresholding was used to assess scar size and transmurality and compared to histology. Relaxation times and contrast-to-noise ratio were compared in healthy and remote myocardium at 1 and 4\u00a0weeks. Linear regression and Bland-Altman was performed to compare infarct size and transmurality.Ten Yorkshire swine underwent 90\u00a0min of occlusion of the circumflex artery and reperfusion. T1\u03c1, T2 and native T1 maps and late gadolinium enhanced (LGE) cardiovascular magnetic resonance (CMR) data were collected at 1\u00a0week (T1\u2009=\u20097.0\u2009\u00b1\u20093.5 (1\u00a0week) and 6.9\u2009\u00b1\u20092.4 (4\u00a0week), CNRT1\u03c1\u2009=\u200912.0\u2009\u00b1\u20096.2 and 12.3\u2009\u00b1\u20093.2, and CNRT2\u2009=\u20098.0\u2009\u00b1\u20093.6 and 10.3\u2009\u00b1\u20095.8. Infarct size was not significantly different for T1\u03c1, T1 and T2 compared to LGE (p\u2009=\u20090.14) and significantly decreased from 1 to 4\u00a0weeks (p\u2009<\u20090.01). Individual infarct size changes were \u2206T1\u03c1\u2009=\u2009\u22123.8%, \u2206T1\u2009=\u2009\u22123.5% and \u2206LGE\u2009=\u2009\u22122.8% from 1 \u2013 4\u00a0weeks, but there was no observed change in infarct size for T2 or histologically.Relaxation time differences between infarcted and remote myocardial tissue were \u2206T1 (infarct-remote)\u2009=\u2009421.3\u2009\u00b1\u2009108.8 (1\u00a0week) and 480.0\u2009\u00b1\u200933.2\u00a0ms (4\u00a0week), \u2206T1\u03c1\u2009=\u200968.1\u2009\u00b1\u200911.6 and 74.3\u2009\u00b1\u200914.2, and \u2206T2\u2009=\u200951.0\u2009\u00b1\u200910.1 and 59.2\u2009\u00b1\u200911.4\u00a0ms. Contrast-to-noise ratio was CNRT1\u03c1 was highly correlated with alterations left ventricle (LV) pathology at 1 and 4\u00a0weeks post-MI and therefore it may be a useful method endogenous contrast imaging of infarction.The online version of this article (doi:10.1186/s12968-017-0332-z) contains supplementary material, which is available to authorized users. Ischemic heart disease is an enormous health and economic burden and the most common cause of death throughout the world . A devasT1\u03c1 (\u201cT-one-rho\u201d) CMR has recently emerged as an endogenous contrast method for quantitative imaging of myocardial injury \u201313. T1\u03c1 We conducted a serial study of reperfused infarction in pigs to monitor T1\u03c1 at 1 and 4\u00a0weeks post-MI. The objectives were to compare T1\u03c1 relaxation time changes in ischemic tissue with native T1, T2 and LGE and associate each with infarct size and transmurality. Finally, imaging results were correlated with fibrosis using histological data.n\u2009=\u200910) were procured for this study. During all procedures, sedation was induced with intramuscular ketamine, endotracheal intubation was performed, and the animal was maintained with a mixture of isoflurane 1-2% and oxygen with a ventilator tidal volume of 20\u00a0mL/kg . Anesthesia and animal temperature was closely monitored for the duration of surgical and imaging procedures to maintain a constant physiologic state. Arterial access was obtained at the carotid artery for measurement of intraventricular pressure . Venous access was obtained at the internal jugular veins for administration of medication. After each procedure, the animal was weaned from anesthesia and transported to the recovery room. Upon completion of the terminal CMR study, the animal was returned to the operating room for euthanasia and tissue harvest.Yorkshire swine (n\u2009=\u20095) and II, a 4-week terminal study (n\u2009=\u20095). Furthermore, five animals underwent a baseline CMR immediately prior to coronary artery occlusion. The experimental model was chosen to emulate human post-percutaneous coronary intervention ischemia injury. All the animals of the 4\u00a0week group underwent both 1 and 4\u00a0week CMR. Figure\u00a0Ten pigs underwent 90\u00a0min of coronary artery occlusion and were randomized into two groups: group I, a 1-week post-MI terminal study with 40\u00a0mT/m gradient and 12 channel RF receiver arrays. Intraventricular pressure was interfaced to physiological monitoring software and filtered to facilitate dual respiratory and cardiac gating . All 2D images were acquired in the short-axis during breath-holding and 3D with dual cardiac and respiratory gating. Breath-holding was performed by temporarily disabling the animal ventilator.x \u2013 SLy - 180y \u2013 SL-y - 90-x) [1\u2009=\u2009500\u00a0Hz, spatial resolution\u2009=\u20091.4\u2009\u00d7\u20091.4\u00a0mm2, slice thickness\u2009=\u20096\u00a0mm, flip angle\u2009=\u200970\u00b0, TE\u2009=\u20091.45\u00a0ms, TR\u2009=\u20092.9\u00a0ms, NSeg\u2009=\u200955, bandwidth\u2009=\u2009900\u00a0Hz/pixel, linear k-space phase encoding ordering, parallel imaging with acceleration factor\u2009=\u20092, 34 reference k-space lines obtained in a separate heartbeat, and four heartbeats for T1 relaxation between shots. The T1\u03c1 amplitude was set at the highest available within scanner specific absorption rate limits (B1\u2009=\u2009500\u00a0Hz).2D T1\u03c1 single-shot balanced steady-state free precession (bSSFP) sequences were performed using a spin echo, spin lock (SL) T1\u03c1 pulse cluster (90 - 90-x) , 15. T1\u03c1x \u2013 180y \u2013 90-x) bSSFP sequence and 8 images were acquired with different contrast times TE\u2009=\u20092, 10, 18, 26, 34, 42, 50\u00a0ms using the same readout as with the T1\u03c1 images.2D T2 maps were obtained using a single-shot T2 prepared . Other p2, slice thickness\u2009=\u20096\u00a0mm.Retrospective, short axis, multi-slice cine CMR was performed with a temporal resolution\u2009=\u200940\u00a0ms, flip angle\u2009=\u200970\u00b0, bandwidth\u2009=\u2009940\u00a0Hz/pixel, spatial resolution\u2009=\u20091.1\u2009\u00d7\u20091.1\u00a0mm2, flip angle\u2009=\u200950\u00b0, TE\u2009=\u20091.6\u00a0ms, TR\u2009=\u20093.2\u00a0ms. slice thickness\u2009=\u20092\u00a0mm, and parallel imaging acceleration factor\u2009=\u20092 [The animals received a 0.1\u00a0mmol/kg intravenous injection of gadolinium contrast for LGE imaging . Imaging was performed 10\u00a0minutes after injection of contrast agent using an inversion time (TI) scout sequence to determine the inversion time to null myocardial tissue signal. LGE CMR was obtained using a 3D multishot phase-sensitive inversion recovery (PSIR) bSSFP sequence at spatial resolution\u2009=\u20091.2\u2009\u00d7\u20091.2\u00a0mmctor\u2009=\u20092 .After ex vivo CMR, the heart was flash-frozen using liquid nitrogen in a 4.5\u00a0L cryogenic container . The heart was sectioned into uniformly thick short axis slices using a commercial-grade prosciutto slicer obtaining slices of uniform thickness of approximately 2\u00a0mm to be matched to CMR studies. Slices were submerged in a PBS solution with 0.1\u00a0M triphenyl tetrazolium chloride (TTC) and incubated at 50\u00a0\u00b0 C for 15\u00a0min . Slices Cine CMR image series were used to calculate indexed LV mass (Mass), wall thickness (WT), end-diastolic volume (EDV), end-systolic volume (ESV), ejection fraction (EF) and cardiac output (CO). Epi- and endocardial contours were drawn manually at ED and ES (excluding papillary muscles) using standard techniques , 20.For all images, scar size was assessed using full width at half maximum (FWHM) thresholding in a mid-LV slice with visible enhancement on LGE (QMass) . FWHM thScar transmurality was computed as the ratio of hyperintense (infarct) to non-enhanced myocardium (%) in a mid-ventricular short axis slice. The myocardium was divided into six circumferential wedges and transmurality was reported for anterolateral and posterolateral segments in which infarction was observed (two of six segments). Scar transmurality <5% in a segment was excluded as noise.Motion correction was used to align T1\u03c1 and T2 images using optical flow estimation of the image deformations . T1 and \u2206TX=TXinf\u2013TXrem and TXinf (TXrem) was the mean relaxation time in the infarcted (remote) tissue and \u03c3(TXrem) was the standard deviation of the relaxation times observed in remote myocardium [Contrast-to-noise ratio (CNR) was calculated for T1\u03c1, T2 and native T1:ocardium .t-test.Descriptive statistics were reported as mean\u2009\u00b1\u2009standard deviation (SD). Comparison of statistical means was performed using 2-way analysis of variance. Correlations were assessed with Pearson\u2019s r. Bland-Altman testing was performed to test for mean bias and variation between imaging methods . Pairwiss\u2009=\u200910) survived the 90\u00a0min ischemia and reperfusion study to their terminal CMR at 1 or 4\u00a0weeks. At 4\u00a0weeks post-MI, there was a 13.4\u2009\u00b1\u20095.4% reduction in EF compared to baseline and associated with a 22.7\u2009\u00b1\u20098.0\u00a0mL increase in ESV and a 24.2\u2009\u00b1\u200914.1\u00a0mL increase in EDV. End diastolic wall thickness decreased by 4.0% after 4\u00a0weeks (p\u2009<\u20090.05) and \u0394WT and \u0394LVEDV between baseline and 4\u00a0weeks were correlated . Additional details are reported in Table\u00a0All pigs s\u2009=\u20090 surviveHyperintense regions were observed at the lateral wall on all T1\u03c1, T2, and T1 maps and in LGE images at 1 and 4\u00a0weeks, consistent with ischemic injury to the circumflex coronary circulation and 12.3\u2009\u00b1\u20093.2 (4\u00a0week), CNRT2\u2009=\u20098.0\u2009\u00b1\u20093.6 and 10.3\u2009\u00b1\u20095.8 and CNRT1\u2009=\u20097.0\u2009\u00b1\u20093.5 and 6.9\u2009\u00b1\u20092.4.As illustrated in Fig.\u00a0ss\u2009=\u20090.14) and, among animals who underwent both 1 and 4\u00a0week CMR, significantly decreased from 1 to 4\u00a0weeks (s\u2009<\u20090.01). Individual infarct size changes were \u0394T1\u03c1\u2009=\u2009\u22123.8%, \u0394T1\u2009=\u2009\u22123.5% and \u0394LGE\u2009=\u2009\u22122.8% from 1 to 4\u00a0weeks , good correlation for T1 and LGE and poor correlation for T2 and LGE and was unchanged from 1 to 4\u00a0weeks (p\u2009=\u20090.15).Infarct transmurality was not significantly different for T1\u03c1, T1 and T2 compared to LGE (p\u2009<\u20090.001), T1 and LGE and moderate correspondence between T2 and LGE T1\u03c1 values had higher relaxation time-dependent change than T2 and contrast-to-noise ratio compared to T1 and T2 in the infarcted myocardium; (2) there was a decrease in infarct size from 1 to 4\u00a0weeks on T1\u03c1, T1 and LGE CMR; and (3) T1\u03c1 infarct size was better correlated with LGE than T1 or T2 and that T2 in particular was poorly correlated. Improved infarct contrast-to-noise ratio on T1\u03c1 may explain the better correlation with LGE and histological infarct size and transmurality than T1 and T2.T1\u03c1 is increased in acute and chronic myocardial infarction , 11, 13,Furthermore, while T2 and native T1 are believed to be biomarkers for myocardial edema in the area-at-risk after acute MI , recent T2 had reduced correlation with LGE as compared to T1\u03c1 and T1, which may be partly explained by differences in the way post-infarction hemorrhage affects endogenous contrast. Degradation of hemoglobin byproducts in the hemorrhage in T2 and T2* CMR studies in patients and animals contribute to increased magnetic susceptibility-induced dephasing on CMR , 28. Acc1. As the amplitude B1 approaches zero, T1\u03c1 approaches T2 . In our comparison of T1\u03c1, T2 and T1, we used identical readout sequences to eliminate a major cause of measurement variation. In addition, we matched the time between adjacent T2 or T1\u03c1 preparations (4 heartbeats) and discarded the first scan because of variations in T1 recovery between preparations.Another reason for the poor T2 agreement with LGE is that different methods of preparation may give different T2, e.g. preparations with adiabatic refocusing pulses instead of a non-selective Hahn spin echo . The freInfarct size is expected to decrease from sub-acute to chronic post-infarction times . T1\u03c1, T1We found that T1\u03c1 and T1 infarct transmurality was correlated with LGE CMR at 1 and 4\u00a0week post-MI. However, differences in T1\u03c1 infarct transmurality might be explained by the varying acquisition times in the cardiac cycle. Although infarct size is not influenced by cardiac cycle, scar transmurality (derived from LGE CMR) has shown to vary between end-diastolic and end-systolic assessment . LGE MR T1\u03c1 CMR is increased in myocardial infarction compared to T2 and has improved contrast-to-noise ratio compared to T1 and T2. Infarct size and transmurality on T1\u03c1 and native T1 endogenous contrast was correlated with LGE CMR and thus may be useful as endogenous contrast CMR methods in ischemic patients who cannot receive contrast agents."} +{"text": "The 1:1, 2-methyl\u00adpyridium and 4-methyl\u00adpyridinium salts of the chiral 4-methyl\u00adbenzo\u00adyloxy-substituted succinic acid form, respectively one- and two-dimensional hydrogen-bonded crystal structures, 6H8N+\u00b7C20H17O8\u2212, comprises a 2-methyl\u00adpyridinium cation and a 2,3-bis\u00ad(4-methyl\u00adbenzo\u00adyloxy)succinate mono-anion while the salt (II), 2C6H8N+\u00b72C20H17O8\u2212\u00b75H2O, consists of a pair of 4-methyl\u00adpyridinium cations and 2,3-bis\u00ad(4-methyl\u00adbenzo\u00adyloxy)succinate mono-anions and five water mol\u00adecules of solvation in the asymmetric unit. In (I), the dihedral angle between the aromatic rings of the anion is 40.41\u2005(15)\u00b0, comparing with 43.0\u2005(3) and 85.7\u2005(2)\u00b0 in the conformationally dissimilar anion mol\u00adecules in (II). The pyridine ring of the cation in (I) is inclined at 23.64\u2005(16) and 42.69\u2005(17)\u00b0 to the two benzene moieties of the anion. In (II), these comparative values are 4.7\u2005(3), 43.5\u2005(3)\u00b0 and 43.5\u2005(3), 73.1\u2005(3)\u00b0 for the two associated cation and anion pairs. The crystal packing of (I) is stabilized by inter-ionic N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds as well as weak C\u2014H\u22ef\u03c0 inter\u00adactions, linking the ions into infinite chains along [100]. In the crystal packing of (II), the anions and cations are also linked by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds involving also the water mol\u00adecules, giving a two-dimensional network across (001). The crystal structure is also stabilized by weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions.The title salt (I), C A white precipitate was formed, which was dissolved in 30\u2005ml of water and then kept at room temperature for slow evaporation. After 2 months, crystals of (I)The title salts (I)Uiso = 1.5Ueq(methyl C or O) and Uiso = 1.2Ueq(aromatic and methyl\u00adene C). H atoms for NH and OH groups were located in difference-Fourier maps and refined with a distance restraint [N\u2014H = 0.86\u2005(1)\u2005\u00c5 or O\u2014H = 0.82\u2005(1)\u2005\u00c5]. The Flack absolute structure obtained for both structures [(for (I)] and 0.6\u2005(3) [for (II)], for 1335 and 2690 quotients, respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017012981/zs2388sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989017012981/zs2388Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017012981/zs2388IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989017012981/zs2388Isup4.cmlSupporting information file. DOI: 1573939, 1573938CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal of the methyl\u00adphenyl compound, the mol\u00adecules are linked 28H24O6, (I), C34H22O5S, (II), and C27H20O3S2, (III), the cyclo\u00adhexa-1,3-diene rings of the 1,2-di\u00adhydro\u00adnaphthalene ring systems adopt half-chair, boat and half-chair conformations, respectively. The carbonyl O atoms attached to the di\u00adhydro\u00adnaphthalene ring systems are each significantly deviated from the mean plane of the 1,2-di\u00adhydro\u00adnaphthalene ring system, by 0.6162\u2005(12)\u2005\u00c5 in (I), 0.6016\u2005(16)\u2005\u00c5 in (II) and 0.515\u2005(3)\u2005\u00c5 in (III). The mean planes of the 1,2-di\u00adhydro\u00adnaphthalene ring systems make dihedral angles of 85.83\u2005(3), 88.19\u2005(3) and 81.67\u2005(8)\u00b0, respectively, with the methyl\u00adphenyl ring in (I), the pyrene ring in (II) and the phenyl ring in (III). In (I), the mol\u00adecular structure is stabilized by an intra\u00admolecular C\u2014H\u22efO hydrogen bond, generating an S(6) ring motif. In the crystal of (I), mol\u00adecules are linked by an inter\u00admolecular C\u2014H\u22efO hydrogen bond, which generates a C(8) zigzag chain running along [100]. Adjacent chains are further connected by C\u2014H\u22ef\u03c0 and offset \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.6572\u2005(9)\u2005\u00c5], forming a double-chain structure. In the crystals of (II) and (III), mol\u00adecules are linked into chain structures by offset \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances of 3.5349\u2005(12) and 3.8845\u2005(13)\u2005\u00c5 for (II) and 3.588\u2005(2)\u2005\u00c5 for (III). In (II) and (III), the thio\u00adphene rings are orientationally disordered over two sites, with occupancy ratios of 0.69:0.31 for (II), and 0.528\u2005(4):0.472\u2005(4) and 0.632\u2005(5):0.368\u2005(5) for (III).In the title 1-oxo-1,2-di\u00adhydro\u00adnaphthalene derivatives, C In (II)via C\u2014H\u22efO hydrogen bonds zigzag chain along to [100]. Adjacent chains are further linked into a double-chain structure \u2005\u00c5, inter\u00adplanar distance = 3.443\u2005(1)\u2005\u00c5, slippage = 1.232\u2005\u00c5; Cg1 and Cg4 are the centroids of the C1\u2013C6 and C22\u2013C27 benzene rings, respectively].In the crystal of compound (I)re Fig.\u00a04 through Cg3\u22efCg6iii = 3.5349\u2005(12)\u2005\u00c5, inter\u00adplanar distance = 3.466\u2005(1)\u2005\u00c5; Cg3\u22efCg7iii = 3.8845\u2005(13)\u2005\u00c5, inter\u00adplanar distance = 3.468\u2005(1)\u2005\u00c5; Cg3, Cg6 and Cg7 are the centroids of the C1\u2013C6, C22\u2013C25/C33/C34 and C25\u2013C29/C34 benzene rings, respectively; symmetry code: (iii) \u2212x, 1/2-y, \u2212z; Fig.\u00a05Cg5\u22efCg7iv = 3.888\u2005(2)\u2005\u00c5, inter\u00adplanar distance = 3.632\u2005(1)\u2005\u00c5; Cg5 and Cg7 are the centroids of the benzene C1\u2013C6 and C22\u2013C27 rings, respectively; symmetry code: (iv) x, 3/2-y, z; Fig.\u00a06In the crystal of (II)Compound (I): To a stirred solution of 1-(4-meth\u00adoxy\u00adphen\u00adyl)-3-p-tolyl\u00adisobenzo\u00adfuran in dry di\u00adchloro\u00admethane (DCM), dimethyl acetyl\u00adenedi\u00adcarboxyl\u00adate (DMAD) was added and the reaction mixture was stirred at room temperature for 1\u2005h. Removal of the solvent was followed by column chromatographic purification gave the isobenzo\u00adfuran\u2013DMAD adduct as a colorless solid . To a stirred solution of isobenzo\u00adfuran\u2013DMAD adduct in dry DCM, BF3\u00b7OEt2 was added and the reaction mixture was stirred at room temperature for 5\u2005min. Removal of the solvent followed by column chromatographic purification gave compound (I)Compound (II): To a stirred solution of 1-(pyren-1-yl)-3-(thio\u00adphen-2-yl)isobenzo\u00adfuran in dry DCM (10\u2005ml), DMAD was added and the reaction mixture was stirred at room temperature for 1\u2005h. To this, BF3\u00b7OEt2 was added and stirred at room temperature for 5\u2005min. Removal of the solvent followed by column chromatographic purification afforded compound (II)Compound (III): To a solution of 1,3-di(thio\u00adphen-2-yl)isobenzo furan in dry toluene (15\u2005ml), ethyl-3-phenyl\u00adpropiolate was added and refluxed till the consumption of 1,3-di(thio\u00adphen-2-yl)isobenzo\u00adfuran (disappearance of fluorescent colour in 8\u2005h). After removal of toluene in vacuo, the crude adduct was dissolved in dry DCM (15\u2005ml), BF3\u00b7OEt2 was added and the reaction mixture was stirred for 10\u2005min at room temperature. Removal of the solvent was followed by column chromatographic purification which afforded compound (III)Uiso(H) values were set to 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms. In compound (II)SIMU and DELU) and bond length restraints (DFIX) with C\u2014S = 1.70\u2005(1)\u2005\u00c5, C\u2014C = 1.50\u2005(1)\u2005\u00c5 and C=C = 1.40\u2005(1)\u2005\u00c5 were applied to the disordered rings.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017000469/is5470sup1.cifCrystal structure: contains datablock(s) I, II, III, global. DOI: 10.1107/S2056989017000469/is5470Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989017000469/is5470IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989017000469/is5470IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989017000469/is5470Isup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989017000469/is5470IIsup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989017000469/is5470IIIsup7.cmlSupporting information file. DOI: 997379, 1438209, 1438503CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit contains two independent mol\u00adecules, which are linked by an O\u2014H\u22efO hydrogen bond. The dimers are further assembled into one-dimensional ladder like structure through O\u2014H\u22efF hydrogen bonds and stabilized by \u03c0\u2013\u03c0 inter\u00adactions. The ladders are further linked by C\u2014H\u22ef\u03c0 contacts. 2 complex, C12H12BF2N3O, contains two independent mol\u00adecules, which are linked by an O\u2014H\u22efO hydrogen bond. The dimers are further assembled into a one-dimensional ladder-like structure through O\u2014H\u22efF hydrogen bonds and stabilized by \u03c0\u2013\u03c0 inter\u00adactions. The ladders are further linked by C\u2014H\u22ef\u03c0 contacts.The asymmetric unit of the title azo\u00adpyrrole-BF Li et al. (20092\u2013azo\u00adpyrrole complexes as sensitizers for dye-sensitized solar cells (DSSCs) have been evaluated and 1.382\u2005(3)\u2005\u00c5, respectively; Table\u00a01pyrrole bonds are shorter than the B\u2014Nazo bonds. The two N\u2014N bonds each adopt a trans conformation and at 1.318\u2005(3) and 1.312\u2005(3)\u2005\u00c5 are much longer than that in the structure of the free azo\u00adpyrrole ligand = 3.544\u2005(1)\u2005\u00c5, Cg2\u22efCg3 = 3.617\u2005(1)\u2005\u00c5 and Cg3\u22efCg4 = 3.664\u2005(13)\u2005\u00c5; Cg1, Cg2, Cg3 and Cg4 are the centroids of the N1/C1\u2013C4, C7\u2013C12 and C19\u2013C24 rings, respectively]. The ladders assemble into a layer structure through C\u2014H\u22ef\u03c0 contacts and 1.310\u2005(1)\u2005\u00c5] are comparable to those in the title compound.A search in the Cambridge Structural Database and tri\u00adethyl\u00adamine (6\u2005mL) in dry di\u00adchloro\u00admethane (15\u2005mL) was slowly added boron trifluoride ethyl ether (2\u2005mL). The resulting solution was stirred for 40\u2005min, and then saturated potassium carbonate solution was added and stirred for 30 minutes. The resulting solution was extracted with ethyl acetate (10\u2005mL \u00d7 3) and evaporated under vacuum to dryness. The residue was purified by column chromatography, eluting with ethyl acetate and petroleum ether (v/v = 1:14), to give a dark-green product, m.p. = 405\u2005K. Yield 65%. 1H NMR : \u03b4 10.118 , 7.548\u20137.526 , 6.920\u20136.897, 6.342 , 2.371, 2.314 . Suitable crystals for X-ray diffraction analysis were obtained by the slow evaporation of an CHCl3/CH3OH solution of the title compound.To a solution of 2-(4-hy\u00addroxy\u00adlphenyl\u00adazo)-3,5-dimethyl-1-Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018006229/ex2007sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018006229/ex2007Isup2.hklStructure factors: contains datablock(s) I. DOI: 1839158CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two mol\u00adecules are linked into dimers by a combination of Hg\u22efHg [Hg\u22efHg = 3.6153\u2005(3)\u2005\u00c5] and C\u2014H\u22efCl and C\u2014H\u22ef\u03c0 inter\u00adactions.The structure of C 3N,C1,N\u2032}[bromido/chlorido\u00ad(0.30/0.70)]mercury(II)\u2013{2,6-bis\u00ad[(di\u00admethyl\u00adamino)\u00admeth\u00adyl]phenyl-\u03ba3N,C1,N\u2032}[bromido/chlorido\u00ad(0.24/0.76)]mer\u00adcury(II) (1/1), [HgBr0.30Cl0.70(C12H19N2)]\u00b7[HgBr0.24Cl0.76(C12H19N2)], there are two mol\u00adecules in the asymmetric unit of formula LHgX {L = 2,6-bis\u00ad[(di\u00admethyl\u00adamino)\u00admeth\u00adyl]phenyl and X = Cl/Br}. In each mol\u00adecule, the halide site is mixed Cl/Br, with occupancies of 0.699\u2005(7):0.301\u2005(7) and 0.763\u2005(7):0.237\u2005(7), respectively. The two mol\u00adecules are linked into dimers by a combination of Hg\u22efHg [Hg\u22efHg = 3.6153\u2005(3)\u2005\u00c5] and C\u2014H\u22efCl and C\u2014H\u22ef\u03c0 inter\u00adactions.In the mol\u00adecular structure of the title compound, {2,6-bis\u00ad[(di\u00admethyl\u00adamino)\u00admeth\u00adyl]phenyl-\u03ba R2Hg and RHgX have received considerable attention in the last three decades, mainly related to the search for versatile reagents in controlled transmetallation reactions :0.237\u2005(7) in mol\u00adecule B. In these moieties, there are two coordination spheres around each Hg atom \u2005\u00c5 is significantly smaller than the sum of the van der Waals radii for Hg\u22efHg 2 dimer at 3.41\u2005\u00c5 \u00admeth\u00adyl]-6-[(di\u00admethyl\u00adamino)\u00admeth\u00adyl]benzene}, which is a dimer linked by Hg2Br2 units, with one di\u00admethyl\u00adamino arm of each ligand coordinated to an Hg atom, and where there are no Hg\u22efHg inter\u00adactions present /2.867\u2005(6)\u2005\u00c5] are significantly shorter than the sum of the van der Waals radii for Hg and N [\u03a3rvdW = 3.53\u2005\u00c5]. However, these values are slightly longer than related organo\u00admercury(II) compounds with one pendant arm of 2.65\u2005(1), 2.725\u2005(4) and 2.647\u2005(2)\u2005\u00c5 \u2005\u00c5, which is significantly smaller than the sum of the van der Waals radii for Hg\u22efHg 2C6H3Br], 1, was prepared according to the procedure given by van Koten and co-workers was employed instead of 2.2 equivalents to quench with 2-bromo-1,3-bis\u00ad(bromo\u00admeth\u00adyl)benzene. This afforded a yellow oil which was purified by vacuum distillation to give a colorless oil in 70% yield. n-BuLi was added dropwise via syringe to the solution of 1 in dry Et2O (15\u2005ml) under an inert atmosphere at 273\u2005K. After 30\u2005min, the color of the reaction mixture changed from colorless to pale yellow. To this, a solution of HgCl2 in dry THF (10\u2005ml) was added. The whole mixture was stirred for 5\u2005h at 273\u2005K and then allowed to warm slowly to room temperature. Then reaction mixture was filtered and the filtrate evaporated to dryness and the resulting precipitate extracted with hexane. The workup afforded a white precipitate of 2 . Colorless crystals of 2 suitable for single-crystal diffraction analysis were obtained by slow diffusion of hexane into CHCl3 at room temperature.The precursor 1H NMR: \u03b4 7.15 , 7.07 , 3.45 , 2.21 . 13C NMR: \u03b4 144.90, 128.36, 128.10, 66.01, 44.85. 199Hg NMR: \u03b4 \u2212930. Analysis calculated for C12H19ClHgN2: C 33.73, H 4.48, N 6.56%; found: C 32.55, H 5.10, N 5.26%. ESI\u2013MS (positive mode): [M + H]+m/z = 429.1005 (observed), 429.1015 .Uiso(H) = xUeq(C), where x = 1.5 for methyl H atoms and 1.2 for all other C-bound H atoms. There are two mol\u00adecules in the asymmetric unit and in each the halide site is occupied by a mix or Cl and Br, with refined occupancies of 0.699\u2005(7):0.301\u2005(7) and 0.763\u2005(7):0.237\u2005(7), respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989017014682/hg5496sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017014682/hg5496Isup2.hklStructure factors: contains datablock(s) I. DOI: 1510433CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, pairs of mol\u00adecules are linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming inversion dimers with graph-set notation 16H13Cl2N3O3S2, the thio\u00adphene ring is disordered in a 0.762\u2005(3):0.238\u2005(3) ratio by an approximate 180\u00b0 rotation of the ring around the S\u2014C bond linking the ring to the sulfonyl unit. The di\u00adchloro\u00adbenzene group is also disordered over two sets of sites with the same occupancy ratio. The mol\u00adecular conformation is stabilized by intra\u00admolecular C\u2014H\u22efCl and C\u2014H\u22efN hydrogen bonds, forming rings with graph-set notation S(5). In the crystal, pairs of mol\u00adecules are linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming inversion dimers with graph-set notation R22(8) and R12(11), which are connected by C\u2014H\u22efO hydrogen-bonding inter\u00adactions into ribbons parallel to (100). The ribbons are further connected into a three-dimensional network by C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions between benzene and thio\u00adphene rings, with centroid-to-centroid distances of 3.865\u2005(2), 3.867\u2005(7) and 3.853\u2005(2)\u2005\u00c5. Hirshfeld surface analysis has been used to confirm and qu\u00adantify the supra\u00admolecular inter\u00adactions.In the title compound, C The mol\u00adecular conformation is stabilized by intra\u00admolecular C\u2014H\u22efCl and C\u2014H\u22efN hydrogen bonds (Table\u00a01S(5).In the mol\u00adecule of the title compound Fig.\u00a01, the dihs Table\u00a01, formingCg1\u22efCg1v, 3.867\u2005(7)\u2005\u00c5 for Cg2\u22efCg2v and 3.853\u2005(2)\u2005\u00c5 for Cg4\u22efCg4vi where Cg1, Cg2 and Cg4 are the centroids of the thio\u00adphene ring B, the thio\u00adphene ring B\u2032 and the benzene ring C .In the crystal, pairs of mol\u00adecules are linked by inter\u00admolecular N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds Table\u00a01 and 3 \u25b8,CrystalExplorer17.5 and external (revdw) atom of the surface:The inter\u00admolecular distance information on the surface can be condensed into a two-dimensional histogram of dnorm= (di \u2212 rivdw)/rivdw + (de \u2212 revdw)/ervdw.dnorm surfaces mapped over a fixed colour scale of \u22121.9033 (red) to 1.1934 (blue). In the fingerprint plots correspond to C\u22efCl (1.3%), N\u22efC (1.3%) and other less important inter\u00adactions (<1%). C\u22efC contacts correspond to inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions. The occurrence of non-high inter\u00adaction rates can be attributed to the fact that the small disordered portion of the mol\u00adecule is not considered.The mol\u00adecular Hirshfeld surfaces were obtained using a standard (high) surface resolution with the three-dimentional trans-rac-methyl 4-methyl\u00adbenzene\u00adsulfonate thio\u00adphen-2-yl]-1,3,4-oxa\u00addiazole thio\u00adphen-2-yl]-1,3,4-oxa\u00addiazole in di\u00adchloro\u00admethane (DCM) were added 2-thio\u00adphene\u00adsulfonamide , 1-ethyl-3-(3-di\u00admethyl\u00adamino-prop\u00adyl)carbodi\u00adimide and 4-dimethyl-amino\u00adpyridine , and the resulting mixture was stirred overnight at room temperature. Upon completion of the reaction, the reaction mixture was partitioned between DCM and water. The collected organic layer was dried over anhydrous Na2SO4, filtered and evaporated to give the crude compound, which was purified with automated-flash chromatography . The obtained product was recrystallized from hexane and ethyl acetate (4:1), m.p. 464.8\u2013465.3\u2005K. 1H NMR (CDCl3): \u03b4 2.24 , 5.33 , 6.64 , 7.12 , 7.21 , 7.45 , 7.69 , 7.97 , 9.29 ; 13C NMR (CDCl3): 11.2, 50.5, 107.6, 127.3, 127.9, 129.1, 129.6, 131.7, 132.9, 133.8, 134.8, 135.1, 139.2, 142.0, 143.5, 158.4. HRMS m/z calculated for C16H13Cl2N3O3S2 [M + H]+ 429.9854; found: 429.9857.To a solution of methyl 1--5-methyl-1Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C) for methyl H atoms. A rotating model was used for the methyl group. The nitro\u00adgen-bound H atom (H1N) was located in a difference-Fourier map and refined with the constraint N1\u2014H1N = 0.84\u2005(3)\u2005\u00c5 and Uiso(H) = 1.2Ueq(N). The thio\u00adphene ring is rotationally disordered by approximately 180\u00b0 over two positions, the ratio of refined occupancies being 0.762\u2005(3):0.238\u2005(3). The di\u00adchloro\u00adbenzene group of the title compound is also disordered over two sets of sites with the same occupancy ratio. The disordered dicholoro\u00adbenzene groups (C: C11\u2013C16 and C\u2032: C11A\u2013C16A) were refined as rigid hexa\u00adgons with bond lengths of 1.39\u2005\u00c5. The displacement ellipsoids for the corresponding carbon atoms in the disordered dicholoro\u00adbenzene groups were constrained by using the EADP command. Six outliers global, I. DOI: 10.1107/S2056989018006242/rz5232Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018006242/rz5232Isup4.cmlSupporting information file. DOI: 10.1107/S2056989018006242/rz5232sup4.pdfCheckcif Report. DOI: 1839201CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the title compound contains two independent mol\u00adecules with very similar conformations. Each mol\u00adecule is built up from a ferrocene unit substituted in the 1 and 1\u2032 positions by a protected sulfur di\u00adphenyl\u00adphosphine and by a di\u00admethyl\u00adhydrazine fragment. 8H11N2)(C17H14PS)], contains two independent mol\u00adecules (A and B) with very similar conformations. Each mol\u00adecule is built up from a ferrocene unit substituted in the 1 and 1\u2032 positions by a protected sulfur di\u00adphenyl\u00adphosphine and by a di\u00admethyl\u00adhydrazine, \u2013C(H)=N\u2014N(CH3)2, fragment. The two independent mol\u00adecules are linked by a C\u2014H\u22efN hydrogen bond. In the crystal, the A\u2013B dimer is linked by a pair of C\u2014H\u22efS hydrogen bonds, forming a centrosymmetric four-mol\u00adecule arrangement. These units are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming a supra\u00admolecular three-dimensional structure.The asymmetric unit of the title compound, [Fe(C The two independent mol\u00adecules are linked by a C\u2014H\u22efN hydrogen bond (Table\u00a02A view of the mol\u00adecular structures of the two independent mol\u00adecules (d Table\u00a02.A and 9.4\u2005(1)\u00b0 for mol\u00adecule B. However, the Cp rings are roughly parallel to each other with a dihedral angle of 1.46\u2005(12)\u00b0 for mol\u00adecule A and 1.85\u2005(12)\u00b0 for mol\u00adecule B. The protected di\u00adphenyl\u00adphosphine and the di\u00admethyl\u00adhydrazine units are approximately trans with respect to the ferrocenyl moiety: the torsion angle P1\u2014C11\u2014C16\u2014C161 and P2\u2014C21\u2014C26\u2014C261 are ca 140.4\u00b0 and ca 141.0\u00b0, respectively.In both mol\u00adecules, the two Cp rings are between eclipsed and staggered conformations, with a twist angle \u03c4 of 5.2\u2005(2)\u00b0 for mol\u00adecule endo towards the FeII ion, by \u22120.7330\u2005(6)\u2005\u00c5 (mol\u00adecule A) and 0.6986\u2005(6)\u2005\u00c5 (mol\u00adecule B) from the Cp ring plane, whereas the phospho\u00adrus atom lies in this plane, displaced by 0.0114\u2005(5) and \u22120.0603\u2005(4)\u2005\u00c5 for mol\u00adecules A and B, respectively. This arrangement, with the protected sulfur atom endo towards the FeII ion and the P atom roughly coplanar with the Cp ring, is quite common in related compounds . The geometry within the hydrazine moiety =N\u2014N(CH3)2 fragments the C=N bond lengths, which vary from 1.270\u2005(5) to 1.287\u2005(4)\u2005\u00c5, and the N\u2014N bond lengths, which vary from 1.367\u2005(4) to 1.382\u2005(5)\u2005\u00c5, are similar to those in the title compound to 0.952\u2005(1)\u2005\u00c5. The corresponding distances for compound 3 fall within these ranges .In ZEQSOD and ZEQSOD02, the P atom is roughly in the Cp ring plane, with deviations from the mean plane ranging from 0.009\u2005(1) to 0.035\u2005(1)\u2005\u00c5, whereas the S atom is 3, is illustrated in Fig.\u00a012), 200\u2005mg (1.66\u2005mmol) of anhydrous magnesium sulfate MgSO4 and 5\u2005ml of anhydrous di\u00adchloro\u00admethane. To the red suspension, 100\u2005ml of N,N-di\u00admethyl\u00adhydrazine was added using a syringe. The reaction mixture was then stirred at room temperature overnight. The crude material obtained was purified by flash chromatography on silica gel to yield 41\u2005mg of compound 3 as a brown solid (yield = 57%). Orange needle-like crystals of 3 were obtained by slow evaporation of a solution in pentane.The synthesis of the title compound, 1H NMR : \u03b4 (p.p.m.): 7.77\u20137.71 , 7.49\u20137.41 , 6.93 , 4.57 , 4.49 , 4.36 , 4.20 , 2.79 . 31P NMR : \u03b4 (p.p.m.): 41.6.Spectroscopic data: Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018000440/su5416sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018000440/su5416Isup2.hklStructure factors: contains datablock(s) I. DOI: 1815294CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Hyperlipidemia, a major pathological condition associated with disrupted lipid levels and physiological redox homeostasis. The excessive release of reactive oxygen species (ROS) leads to enhanced lipid peroxidation, aggravated atherosclerosis and oxidative stress. Integration of natural antioxidant blends in alone or with conventional treatments can alleviate these issues synergistically contributing least side effects. Published literature reported the efficacy of natural antioxidants as individual and in combinations in various conditions but less data is available on their evaluation in low dose ratio blends particularly in hypercholesterolemic diet.n\u2009=\u20096) weighing 1.2\u20131.6\u00a0kg, supplemented with high cholesterol diet (400\u00a0mg/kg) for 12\u00a0weeks were used in the experiment. Antioxidants were administered individual high (100\u00a0mg/kg) and in low dose combinations . Student\u2019s t test and one way analysis of variance (ANOVA) followed by Dunnet\u2019s test were used as statistical tools for evaluation.Antihyperlipidemic effects of selected natural antioxidants; the phenolic oligomeric proanthocyanidins (OPC) and pterostilbene (PT) with niacin (NA) were investigated in current study. Their effects on lipid profile, lipid peroxidation and their aptitude to establish redox state between oxidants and antioxidants in body were evaluated in high cholesterol diet fed animal model. Male albino rabbits (p >0.001) observed in 50:30:20 ratios of OPC, NA and PT, compared to individual therapies 37 and 18\u00a0% max respectively. Moreover the results were also in close proximity with the statin therapy .The results showed synergistic effects of low dose antioxidant blends. Therapies retarded elevation in blood lipid levels, lipid peroxidation and blood antioxidant depletion and consequently contributed in reestablishing redox homeostasis. The LDL/HDL ratio and atherogenic index were suppressed significantly in blend therapies with maximum effects of 59.3 and 25\u00a0% (This study provides an evidence for natural antioxidants blends superiority over individual therapy in chronic diseases like hyperlipidemia. Such therapies in human equivalent doses can help in mitigating chronic illnesses in general populations. Their oxidation leads to severe endothelial cell injury and atherosclerosis [Hyperlipidemia is a major pathological condition associated with mortality and disability in both developed and developing countries . Chronicclerosis \u20136. The sclerosis . Howeverclerosis . The antclerosis , 10. In clerosis , 11, 12.clerosis \u201316.Natural antioxidants were evaluated in individual and in combinations for lipid lowering and free radical scavenging properties and were found effective in depressing the disease progression \u201322. In cOligomeric proanthocyanidins (OPC) is a polyphenolic compound belongs to flavonoids class, widely distributed in plants and posses potent antioxidant and lipit test was applied to evaluate the difference statistically. Lipid profiles are calculated as mean\u2009\u00b1\u2009SEM (n\u2009=\u20096) along with p-values in Table\u00a0Acute rise in lipid profiles were observed in rabbits administered with cholesterol for 2\u00a0weeks compared to healthy group. Almost two to four time rise was observed in every parameter. Student\u2019s p <0.001), 20.26\u2009\u00b1\u20090.94 (p <0.001) and 19.19\u2009\u00b1\u20090.91 (p <0.01) and AI values 0.893\u2009\u00b1\u20090.033 (p <0.01), 0.896\u2009\u00b1\u20090.053 (p <0.01), 0.971\u2009\u00b1\u20090.073 (p <0.05) respectively. The atorvastatin therapy showed 14.0\u2009\u00b1\u20090.99 (p <0.001) and 0.785\u2009\u00b1\u20090.061 (p <0.001) compared to disease control 30.50\u2009\u00b1\u20090.69, 1.096\u2009\u00b1\u20090.040. In combination therapies the results were even more significant and synergism were observed by improving blood lipid profile depending on dose ratio in particular combination. Among combinations of OPC with NA the 70:30 ratio delivered significant effects by suppressing LDL/HDL ratio to 15.36\u2009\u00b1\u20090.98 (p <0.001) while differ nonsignificantly from other two ratios 16.77\u2009\u00b1\u20090.69 (p <0.001) and 17.33\u2009\u00b1\u20090.75 (p <0.001). AI was significantly reduced to 0.843\u2009\u00b1\u20090.041 (p <0.001), 0.837\u2009\u00b1\u20090.045 (p <0.001) and 0.887\u2009\u00b1\u20090.037 (p <0.01) in 70:30, 30:70 and 50:50 respectively. Similarly in OPC with PT combinations all the ratios provided significant effects compared to disease control but no significant differences were observed between the groups. The LDL/HDL and AI were 14.60\u2009\u00b1\u20090.98 (p <0.001) and 0.851\u2009\u00b1\u20090.041 (p <0.001) in 70:30 ratio, 15.19\u2009\u00b1\u20090.42 (p <0.001), 0.929\u2009\u00b1\u20090.057 (p <0.05) in 30:70 and 16.33\u2009\u00b1\u20090.75 (p <0.001), 0.906\u2009\u00b1\u20090.037 (p <0.01) in 50:50 respectively. In NA and PT, the 70:30 and 50:50 combinations induced similar effects on LDL/HDL ratios 15.16\u2009\u00b1\u20090.89 and 15.16\u2009\u00b1\u20090.84 (p <0.001) respectively, while comparing AI values results were more profound in 70:30 (0.858\u2009\u00b1\u20090.053) (p <0.001) compared to other groups 0.919\u2009\u00b1\u20090.037 (p <0.05) and 0.868\u2009\u00b1\u20090.037 (p <0.01) respectively. In general the two drug combinations showed synergism, evident from significant outcome however differ less from one another [Single drug therapies showed significant effects in reducing lipid levels, confirmed by LDL/HDL ratio and atherogenic index (AI). The OPC, NA and PT maintained LDL/HDL ratios by 19.49\u2009\u00b1\u20090.87 (p <0.001) comparable with the statin outcome 52.66, 26.28\u00a0% (p <0.001) respectively. Though there were minor differences among the groups but differ significantly from the disease control. The high dose single drug therapies suppressed LDL/HDL by 37\u00a0% and AI by 18.5\u00a0% maximum, which are much less as compared to low dose blend therapies confirmed the superiority of combinational therapies. Lipid profiles in mean\u2009\u00b1\u2009SEM and %change in blood lipids at 95\u00a0% confidence interval are summarized in Tables\u00a0The ratios with optimum lipid lowering effects in above therapies were subsequently molded in to three drug combinations to evaluate the extent of synergism of antioxidants acting through various mechanisms. The antioxidant with highest ratio (70\u00a0%) was reduced by 20\u00a0mg and this amount was replaced with third candidate. All three drug combinations showed more significant results as compared to two drug combinations. Most significant effects were observed in 50:30:20 ratio of OPC, NA and PT which suppressed LDL/HDL ratio by 59.3\u00a0% and AI by 25\u00a0% respectively in combination therapies compared to disease control 19.30\u2009\u00b1\u20091.18, 3.01\u2009\u00b1\u20090.28 and 3.73\u2009\u00b1\u20090.412\u00a0\u03bcmol/L. Although in individual therapies the raised levels of methionine, GSH and NAC (24.73\u2009\u00b1\u20091.04\u00a0\u03bcmol/L (p <0.05), 3.38\u2009\u00b1\u20090.42\u00a0\u03bcmol/L (p <0.01) and 3.86\u2009\u00b1\u20090.404\u00a0\u03bcmol/L (p <0.05)) were less significant compared to combination therapies. The all-trans retinoic acid showed opposite results and increased parallel with the lipid levels from 1.80\u2009\u00b1\u20090.10\u00a0\u03bcmol/L (blank control) to 4.48\u2009\u00b1\u20090.24\u00a0\u03bcmol/L (disease control) while in treatment groups its elevation was retarded down to 3.26\u2009\u00b1\u20090.12\u00a0\u03bcmol/L (p <0.001) max; observed on 70:30 ratio of OPC and PT.Significant up-regulation was observed in levels of methionine, GSH and NAC upto 35.08\u2009\u00b1\u20091.60, 3.82\u2009\u00b1\u20090.47 and 4.21\u2009\u00b1\u20090.425\u00a0\u03bcmol/L (p <0.01) among individual therapies and more in combinational therapies ranging 2.51\u2009\u00b1\u20090.092 to 3.40\u2009\u00b1\u20090.097 (p <0.001), compared to disease control 2.12\u2009\u00b1\u20090.058. Homocysteine level was significantly controlled down to 10.09\u2009\u00b1\u20091.39\u00a0\u03bcmol/L (p <0.01), although nonsignificant variations were observed among treatment groups. Almost half of groups delivered nonsignificant effects on ascorbic acid levels with maximum effect of 5.29\u2009\u00b1\u20090.51\u00a0\u03bcmol/L (p <0.001) observed in 50:30:20 ratio of OPC, NA and PT compared to disease control 4.90\u2009\u00b1\u20090.59\u00a0\u03bcmol/L. The natural antioxidants were able to correct the antioxidant levels in blood and are more effective when used in blend therapies rather than individual as summarized in mean\u2009\u00b1\u2009SEM in Tables\u00a0The improvement in GSH/GSSG ratios was significant in OPC (2.83\u2009\u00b1\u20090.065) (\u226599\u00a0% pure). Cholesterol kit; TC, TG and HDL (Human diagnostics GmbH Germany). Oligomeric proanthocyanidins (OPC) , pterostilbene (PT) (\u226599\u00a0% pure) and niacin (NA) (Scharlau Chemie SA) (\u226598\u00a0% pure).HPLC system linked with a DECADE II Electrochemical Detector , HPLC column Discovery HS C18 RP chromatographic column . Lambda-25 UV/Visible spectrophotometer (Perkin Elmer). Centrifuge machine (Centurion scientific Ltd) and Incubator (Incucell Med Center GmbH Germany) were used in the analysis.n\u2009=\u20096. Three groups were treated with OPC, PT and NA individually in 100\u00a0mg/kg doses along with cholesterol doses. In 09 groups, low dose combinatorial blends of all the three drugs in 30:70, 50:50 and 70:30 ratios were administered and most effective combination in each two drug combinations was further incorporated in three drug combinations were used in the experiment. The experimental protocol for our study was approved by ethical committee of Department of Pharmacy University of Peshawar under reference number: 04/EC-15/Pharm. After 7\u00a0days of acclimatization in animal house and bio assay center, hyperlipidemia was developed according to the previous reported protocol . AnimalsPlasma lipid profiling was performed on commercial kits. Base line levels were recorded and samples were taken at every 2\u00a0weeks interval after overnight fasting to evaluate the effect of therapies on blood lipid levels sequentially. Total cholesterol (TC), teiglyceride (TG) and high density lipoproteins (HDL-C) were calculated enzymatically, while low density lipoproteins (LDL-C), very low density lipoproteins (VLDL-C) and atherogenic index (AI) were calculated according to the following formulae \u201365.1\\doctrans retinol (ATR) were determined simultaneously using HPLC-UV [Serum levels of water soluble thiol antioxidants; glutathione (GSH) its oxidized form (GSSG), GSH/GSSG ratio, cystine, cysteine (cys), Homocysteine (Hcy), methionine (meth), N-acetylcysteine (NAC) long with ascorbic acid (AA) and lipid peroxidation were determined simultaneously at the end of study using RP-HPLC- electro chemical detection (ECD) method . Fat sol HPLC-UV .t test for pair analysis and one way analysis of variance (ANOVA) followed by Dunnett\u2019s test for comparison of lipid lowering effect in all treatment groups with respect to the disease control (with no treatment). Data is presented in mean\u2009\u00b1\u2009standard error of mean (SEM) at level of significance p <0.05\u00a0%.Data analysis was performed by applying student\u2019s"} +{"text": "II complex containing an \u03c3-dimerized TCNQ\u2013TCNQ unit is presented, with a C\u2014C bond length of 1.653\u2005(11)\u2005\u00c5. In addition, the \u03c3-dimerized TCNQ\u2013TCNQ unit (refined 75% occupancy) is disordered, forming also a less populated pair of TCNQ mol\u00adecules (25% occupancy) with tightly \u03c0-stacked di\u00adcyano\u00admethanide groups.The first example of an Ni 3)2, 2,2\u2032-bipyridine (bpy) and LiTCNQ in a 1:3:2 molar ratio yielded single crystals of bis\u00ad[tris\u00adnickel(II)] bis\u00ad bi hexa\u00adhydrate, [Ni(C10H8N2)3]2(C24H8N8)(C12H4N4)2\u00b76H2O or [Ni(bpy)3]2(TCNQ\u2013TCNQ)(TCNQ)2\u00b76H2O. The crystal structure comprises [Ni(bpy)3]2+ complex cations, two centrosymmetric crystallographically independent TCNQ\u00b7\u2212 anion radicals with \u03c0-stacked exo groups, and an additional dimeric TCNQ\u2013TCNQ unit which comprises 75.3\u2005(9)% of a \u03c3-dimerized (TCNQ\u2013TCNQ)2\u2212 dianion and 24.7\u2005(9)% of two TCNQ\u00b7\u2212 anion radicals with tightly \u03c0-stacked exo groups. The title complex represents the first example of an NiII complex containing a \u03c3-dimerized (TCNQ\u2013TCNQ)2\u2212 dianion. Disordered solvent water mol\u00adecules present in the crystal structure participate in hydrogen-bonding inter\u00adactions.Crystallization from an aqueous methanol system composed of Ni(NO Several complexes of NiII-containing TCNQ species have been reported previously, e.g. [Ni(terpy)2](TCNQ)2 with non-coordinating \u03c0-dimerized anion radicals with \u03c3-coordinating anion radicals 4\u00b7(CH3)2CO was reported, along with the results of its crystal structure analysis 3]2(TCNQ\u2013TCNQ)(TCNQ)2\u00b76H2O (1) and report here its crystal structure.In the quest for new promising mol\u00adecular magnetic materials besides the complexes of 31, comprises two [Ni(bpy)3]2+ complex cations, a centrosymmetric TCNQ\u2013TCNQ dimeric unit, two centrosymmetric crystallographically independent TCNQ\u00b7\u2212 anion radicals, and three crystallographically independent disordered solvent water mol\u00adecules 8]\u00b76H2O :0.247\u2005(9) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 2\u00a0\u2212\u00a0z] dimerization bond length is 1.653\u2005(11)\u2005\u00c5 and this value is within the usual range (see Database survey section). At the same time, this value is longer than a usual single C\u2014C bond and, consequently, the corresponding bond angles around the C37A atom range from 105.6\u2005(4) to 113.6\u2005(3)\u00b0, displaying significant deviations from the ideal tetra\u00adhedral angle. In the less populated pair of anion radicals within TCNQA, the distance between the C37B atom and its symmetry-related counterpart C37Biii is 3.06\u2005(2)\u2005\u00c5; the inter\u00adplanar distance between the least-squares plane P1 formed by atoms C31B, C37B, C38B and C39B and the least-squares plane P2 formed by their symmetry-related counterparts through a centre of symmetry at is 3.03\u2005\u00c5. The distance of the C37iii atom from the plane P1 is 2.90\u2005\u00c5 and the slippage between atoms C37B and C37Biii is 0.98\u2005\u00c5. These geometric parameters suggest a very strong \u03c0-inter\u00adaction between the less populated pair of anion radicals in TCNQA, and they are pre-positioned for \u03c3-dimerization with little structural rearrangement required upon formation of the covalent bond. This could be seen as an indication of \u03c3-bond formation in the solid state upon crystallization rather than pre-formation of the \u03c3-dimers in solution.The unit cell of the title complex, s Figs. 1 \u25b8 \u25b8 \u25b8\u00b7\u2212 anion radicals, TCNQB and TCNQC, in the crystal structure of 1 \u2019 by Ballester et al. (1999Biii and N10vii is, however, much longer than for the \u2018front-end\u2019 di\u00adcyano\u00admethanide group. It is outside the usually observed range for strong \u03c0-stacking inter\u00adactions in analogous systems 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z] and the second shortest contact is 3.479\u2005(4)\u2005\u00c5 between atoms C46 and C53ii; the latter distance is already somewhat longer due to the noncoplanarity of the two anion radicals. These observed distances are at the upper border for stacking arrangements reported for similar compounds hydrogen-bonded bridges involving TCNQA dianions and TCNQC anion radicals, yielding a supra\u00admolecular layer within the bc plane form an additional hydrogen-bonded bridging path, N\u22efH\u2014O2A\u22efH\u2014O3B\u2014H\u22efO1B\u2014H\u22efN, between the supra\u00admolecular layers. On the other hand, the least-occupied position (O2B) of water mol\u00adecule O2 seems to be hydrogen bonded only to the nitrile N atom and so partially occupies the void in the structure in alternation with its symmetry-related atom O2Bxi (bipy)]\u00b7p-xy (TCNQ\u2013TCNQ)] \u2005\u00c5, which is in line with the observed range in the published crystal structures.A search of the CSD (Groom 3)2\u00b76H2O and bpy in methanol (2\u2005ml) at the same temperature. The dark-green solution that resulted was immediately enclosed in a 5\u2005ml vial and cooled to room temperature (8.75\u2005K\u2005h\u22121) in a programmable drying oven. The dark-green crystalline solid that resulted was filtered off, washed with a small amount of methanol and ether, and dried in air. The solid was mainly of microcrystalline character, with a few single crystals suitable for X-ray study (yield 63%). IR : 3341 (m), 3382 (m), 3074 (vw), 3033 (vw), 2200 (s), 2175 (vs), 2152ssh, 1598 (m), 1581 (s), 1504 (s), 1471 (m), 1441 (m), 1359 (s), 1182 (m), 1020 (w), 987 (w), 826 (w), 779 (m), 765 (m), 737 (w), 653 (w), 483 (w). CNH : C 65.54/67.00, H 3.87/3.98, N 19.81/19.80.A solution of LiTCNQ in methanol (2\u2005ml) heated to 323\u2005K was added dropwise to a mixture of Ni2\u2212 dianion is positionally disordered were considered to be exactly one half, while the refined site-occupation factors for atoms O2A and O2B are 0.908\u2005(3) and 0.092\u2005(3), respectively. Some of the water H atoms were resolved in difference maps and all H-atom positions were refined assuming idealized geometric parameters of O\u2014H = 0.85\u2005(1)\u2005\u00c5 and H\u22efH = 1.344\u2005(1)\u2005\u00c5. For the H atoms of the O2B water mol\u00adecule (the least-occupied water mol\u00adecule), a riding model was used. The Uiso parameters for water H atoms were set at 1.5 times the Ueq value of the parent O atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016019162/zl2686sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989016019162/zl2686Isup2.hklStructure factors: contains datablock(s) I. DOI: 1520298CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, complex and DMF solvate mol\u00adecules build up a one-dimensional hydrogen-bonded polymer along [010].The homoleptic nickel\u2013thio\u00adsemicarbazonate complex shows structural features including an unusual II acetate tetra\u00adhydrate with 4-methyl-2-hydrazinecarbo\u00adthio\u00adamide in a 2:1 molar ratio and recrystallization from di\u00admethyl\u00adformamide yielded the title compound, [Ni(C12H14N3S)2]\u00b7C3H7NO. The ligands act as monoanionic \u03ba2N1,S-donors, forming five-membered metallarings. The NiII ion is fourfold coordinated in a distorted square-planar cis-configuration, which is rather uncommon for mono\u00adthio\u00adsemicarbazone complexes. Intra\u00admolecular H\u22efNi trans-inter\u00adactions are observed [H\u22efNi distances are 2.50 and 2.57\u2005\u00c5] and thus anagostic inter\u00adactions can be suggested. The Hirshfeld surface analysis indicates that the major contributions for the crystal packing are H\u22efH (66.6%), H\u22efS (12.3%) and H\u22efC (10.9%) inter\u00adactions. In the crystal, the complex mol\u00adecules are linked by di\u00admethyl\u00adformamide solvent mol\u00adecules through N\u2014H\u22efO inter\u00adactions into one-dimensional hydrogen-bonded polymers along [010].The reaction of Ni The hydrazinic H atom can be easily removed and the negative charge is then delocalized over the C\u2014N\u2014N\u2014C\u2014S backbone, which enables chemical bonding with many different metal ions hydrazinecarbothi\u00adamide as ligand \u2005\u00c5 for C3 and of 0.3685\u2005(15)\u2005\u00c5 for C15. The two ligands are deprotonated with the negative charge delocalized over the C\u2014N\u2014N\u2014C\u2014S entity, as suggested by their inter\u00admediate bond lengths and supported by the sp2-hybridization for C1, C13, N1, C11, C23 and N4. The imine and thio\u00adamide C\u2014N distances indicate considerable double-bond character, while the C\u2014S distance is consistent with mainly single-bond character. The change of the bond lengths is a key feature to distinguish free, i.e. non-coordinating, and coordinating thio\u00adsemicarbazones. For the title compound these distances are C1\u2014N1 = 1.306\u2005(2), N1\u2014N2 = 1.408\u2005(2), N2\u2014C11 = 1.307\u2005(2) and C11\u2014S1 = 1.752\u2005(2) for one ligand and C13\u2014N4 = 1.303\u2005(2), N4\u2014N5 = 1.409\u2005(2), N5\u2014C23 = 1.303\u2005(2) and C23\u2014S2 = 1.7573\u2005(19) for the other one.One mol\u00adecule of the title complex and one di\u00admethyl\u00adformamide solvate comprise the asymmetric unit. The Nids Fig.\u00a01. The maxcis coordination mode, which is rather uncommon for mono-thio\u00adsemicarbazone ligands, as well as two positioned trans H\u22efNi anagostic inter\u00adactions \u2212x\u00a0+\u00a01, y\u00a0\u2212\u00a0z\u00a0+\u00a0In the crystal, the coordination entities are linked by DMF solvate mol\u00adecules through N\u2014H\u22efO inter\u00adactions. The DMF-oxygen atoms are hydrogen-bond acceptors, forming a bridg\u00ading structure between two N\u2014H\u22efO arrangements: N6\u2014H6\u22efO1 and N3\u2014H3\u22efO10] Fig.\u00a03. Additio0] Fig.\u00a03.et al., 2012de (y axis) and di (x axis) values are the closest external and inter\u00adnal distances from given points on the Hirshfeld surface contacts -4-phenyl-hydrazine\u00adcarbo\u00adthio\u00adamidate-\u03ba2N1,S)}nickel(II) monohydrate bis\u00ad(tetra\u00adhydro\u00adfurane) solvate = 1.2Ueq(C and N) for the sp2\u2013hybridized DMF C atom, the aromatic and the secondary C atoms, and for all N atoms, and with Uiso(H) = 1.5Ueq(C) for the methyl C atoms. The bond lengths are: C\u2014H = 0.99 for \u2013CH2\u2013 fragments, C\u2014H = 0.98 for CH3\u2013 fragments, C\u2014H = 0.95 for aromatic groups and the sp2-hybridized DMF C atom; N\u2014H = 0.88 for all N atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017007198/wm5389sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989017007198/wm5389Isup2.hklStructure factors: contains datablock(s) I. DOI: 1550129CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title benzoyl\u00adthio\u00adurea derivative (MPiCB), the piperidine ring has a chair conformation and its mean plane is inclined to the 4-meth\u00adoxy\u00adbenzene ring by 63.0\u2005(3)\u00b0. 14H18N2O2S, the piperidine ring has a chair conformation. Its mean plane is twisted with respect to the 4-meth\u00adoxy\u00adbenzoyl ring, with a dihedral angle of 63.0\u2005(3)\u00b0. The central N\u2014C(=S)\u2014N(H)\u2014C(=O) bridge is twisted with an N\u2014C\u2014N\u2014C torsion angle of 74.8\u2005(6)\u00b0. In the crystal, mol\u00adecules are linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming chains along the c-axis direction. Adjacent chains are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming layers parallel to the ac plane. The layers are linked by offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.927\u2005(3)\u2005\u00c5], forming a supra\u00admolecular three-dimensional structure.In the title compound, C The central N\u2014C(=S)\u2014N(H)\u2014C(=O) bridge is twisted with an N2\u2014C8\u2014N1\u2014C7 torsion angle of 74.8\u2005(6)\u00b0. The meth\u00adoxy group lies in the plane of the benzene ring, with the C14\u2014O2\u2014C4\u2014C3 torsion angle being 180.0\u2005(4)\u00b0.The mol\u00adecular structure of the title compound, MPiCB, is illustrated in Fig.\u00a01c-axis direction (Table\u00a01ac plane (Table\u00a01Cg\u22efCgi = 3.927\u2005(3)\u2005\u00c5; Cg is the centroid of the C1\u2013C6 ring; inter\u00adplanar distance = 3.517\u2005(2)\u2005\u00c5; slippage = 1.747\u2005\u00c5; symmetry code: (i) \u2212x, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a02].In the crystal of MPiCB, neighbouring mol\u00adecules are linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming chains along the n Table\u00a01. Adjacene Table\u00a01. The layet al., 2016N-(carbono\u00adthio\u00adyl)benzamide skeleton gave 37 hits. Two compounds are of particular inter\u00adest, namely 4-meth\u00adoxy-N-(pyrrolidin-1-ylcarbono\u00adthio\u00adyl)benzamide -3,4,5-tri\u00admeth\u00adoxy\u00adbenz\u00adamide benzamide was added slowly to ammonium thio\u00adcyanate (0.01\u2005mol) in acetone and the mixture was stirred for 30\u2005min at room temperature. A white precipitate of ammonium chloride was filtered off and the filtrate was cooled in an ice bath (278\u2013283\u2005K) for about 15\u2005min. A cold solution (278\u2013283\u2005K) of piperidine (0.01\u2005mol) in acetone was added to the benzoyl iso\u00adthio\u00adcyanate and the mixture was left for 3\u2005h at room temperature. A yellowish precipitate was formed, filtered and washed with cold water to give pale-yellow crystals .3) 2900, \u03bd(C=O) 1609, \u03bd(C\u2014Cbenzene) 1460, \u03bd(C\u2014Ostretching) 1327 and v(C=S) 1252\u2005cm\u22121. The 1H NMR spectrum exhibits the H(N) group at 8.35\u2005Hz, while the 13C NMR signal of the C=S and C=O groups appear at 174.66 and 163.19\u2005Hz, respectively.The infrared spectrum of MPiCB shows the characteristic signals for \u03bd(NH) 3300, \u03bd(O\u2014CHUiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017013317/su4162sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989017013317/su4162Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017013317/su4162Isup3.cmlSupporting information file. DOI: 1575129CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II complex mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds into supra\u00admolecular chains propagating along the [110] direction.In the crystal, the title centrosymmetric Zn 8H4NO2)2(C10H14N2O)2(H2O)2], the ZnII cation, located on an inversion centre, is coordinated by two water mol\u00adecules, two 4-cyano\u00adbenzoate (CB) anions and two di\u00adethyl\u00adnicotinamide (DENA) ligands in a distorted N2O4 octa\u00adhedral geometry. In the mol\u00adecule, the dihedral angle between the planar carboxyl\u00adate group and the adjacent benzene ring is 9.50\u2005(14)\u00b0, while the benzene and pyridine rings are oriented at a dihedral angle of 56.99\u2005(5)\u00b0. The water mol\u00adecules exhibit both an intra\u00admolecular hydrogen bond and an inter\u00admolecular hydrogen bond ; the latter lead to the formation of supra\u00admolecular chains propagating along the [110] direction.In the title complex, [Zn(C N,N-di\u00adethyl\u00adnicotinamide (DENA) is an important respiratory stimulant 2(C7H4ClO2)2(H2O)2] 2(C10H14N2O)2(H2O)2] is one form of niacin. A deficiency of this vitamin leads to loss of copper from the body, known as pellagra disease. Victims of pellagra show unusually high serum and urinary copper levels and DENA ligands, namely trans-di\u00adaqua\u00adbis\u00ad(4-cyano\u00adbenzoato-\u03baO)bis\u00adzinc(II), [Zn(DENA)2(CB)2(H2O)2], and report herein its crystal structure.The structure-function\u2013coordination relationships of the aryl\u00adcarboxyl\u00adate ion in ZnII atom located on an inversion centre, one 4-cyano\u00adbenzoate (CB) ligand, one N,N-di\u00adethyl\u00adnicotinamide (DENA) ligand and one water mol\u00adecule, all ligands coordinating to the ZnII atom in a monodentate manner of the two symmetry-related monodentate CB anions and the two symmetry-related water O atoms (O4 and O4i) around the Zn1 atom form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination sphere is completed by the two pyridine N atoms (N2 and N2i) of the two symmetry-related monodentate DENA ligands in the axial positions \u2005\u00c5 below the planar (O1/O2/C1) carboxyl\u00adate group. The O\u2014Zn\u2014O and O\u2013Zn\u2014N bond angles range from 87.64\u2005(5) to 92.36\u2005(5)\u00b0.In the carboxyl\u00adate groups, the C\u2014O bonds for coordinating O atoms are 0.0148\u2005(19)\u2005\u00c5 longer than those of the non-coordinating ones [C1\u2014O1 = 1.2436\u2005(19)\u2005\u00c5 and C1\u2014O2 = 1.2584\u2005(18)\u2005\u00c5], indicating delocalized bonding arrangements rather than localized single and double bonds. The Zn\u2014O bond lengths are 2.1503\u2005(11)\u2005\u00c5 (for water O atoms) and 2.0842\u2005(10)\u2005\u00c5 (for benzoate O atoms) and the Zn\u2014N bond length is 2.1501\u2005(11)\u2005\u00c5, the Zn\u2014O bond lengths for water oxygen atoms are The dihedral angle between the planar carboxyl\u00adate group (O1/O2/C1) and the adjacent benzene ring (C2\u2013C7) is 9.50\u2005(14)\u00b0, while the benzene and pyridine (N2/C9\u2013C14) rings are oriented at a dihedral angle of 56.99\u2005(5)\u00b0.w\u22efOc hydrogen bonds (Table\u00a01S(6) hydrogen-bonding motifs hydrogen bonds in H2O (50\u2005ml) and di\u00adethyl\u00adnicotinamide in H2O (10\u2005ml) with sodium 4-cyano\u00adbenzoate in H2O (100\u2005ml). The mixture was filtered and set aside to crystallize at ambient temperature for several days, giving translucent intense colourless single crystals.The title compound was prepared by the reaction of ZnSO2O) were located in a difference Fourier map and were refined freely. The C-bound H atoms were positioned geometrically with C\u2014H = 0.93, 0.97 and 0.96\u2005\u00c5, for aromatic, methyl\u00adene and methyl H atoms, respectively, and constrained to ride on their parent atoms, with Uiso(H) = k \u00d7 Ueq(C), where k = 1.5 for methyl H atoms and k = 1.2 for aromatic and methyl\u00adene H atoms.The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0210.1107/S2056989016013815/xu5890sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989016013815/xu5890Isup2.hklStructure factors: contains datablock(s) I. DOI: 1501337CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, there are only weak C\u2014H\u22efO hydrogen bonds and offset \u03c0\u2013\u03c0 inter\u00adactions present.The whole mol\u00adecule of the title di\u00adaryl\u00adcarbonate is generated by mirror symmetry, the mirror bis\u00adecting the central benzene ring, and the carbonate groups adopt an 20H14O6, is generated by mirror symmetry, the mirror bis\u00adecting the central benzene ring. The carbonate groups adopt an s-cis-s-cis conformation, with torsion angles of 58.7\u2005(2) and 116.32\u2005(15)\u00b0. The crystal structure of 1,3-phenyl\u00adene bis\u00ad(phenyl carbonate) contains no strong hydrogen bonds, though weak C\u2014H\u22efO and offset \u03c0\u2013\u03c0 inter\u00adactions are observed, forming layers parallel to the ac plane.The whole mol\u00adecule of the title compound, C We surmise this compound formed through a combination of inter\u00admolecular \u2018self-alcoholysis\u2019 reactions leading to a carb\u00adamate inter\u00admediate structure reported herein was identified as an unexpected side product from the attempted recrystallization of 1- and 116.32\u2005(15)\u00b0, respectively. The 1,3-substitution of the central aromatic ring imparts the mol\u00adecule with a bent or \u2018U-shape\u2019 conformation and a significant net dipole moment.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01a-axis direction are related by glide symmetry and assemble into polar chains \u2005\u00c5, inter\u00adplanar distance = 3.438\u2005(1)\u2005\u00c5, with a slippage of 1.669\u2005\u00c5 ; Cg2\u22efCg2iii,iv = 3.822\u2005(1)\u2005\u00c5, inter\u00adplanar distance = 3.398\u2005(1)\u2005\u00c5, with a slippage of 1.749\u2005\u00c5 , C\u2014O bond lengths fall within 1.310 and 1.387\u2005\u00c5 [average: 1.343\u2005(9)\u2005\u00c5], and O\u2014C\u2014O angles average 106\u2005(1)\u00b0. However, torsion angles about the C\u2014O\u2014C\u2014Carom bonds are extremely variable.A search of the Cambridge Structural Database was identified in this search, 4,4\u2032-iso\u00adpropyl\u00adidenediphenyl-bis\u00ad(phenyl\u00adcarbonate) -3-phenyl\u00adurea was filtered, dried, and recrystallized in assorted organic solvents . Slow evaporation of an ethano\u00adlic solution in a 1 dram vial, capped with pierced lids, yielded large colorless plates of 1,3-phenyl\u00adene bis\u00ad(phenyl carbonate). Needle-like crystals identified within the same vials corresponded to 1-(m-phenol)-3-phenyl\u00adurea. The appearance of 1,3-phenyl\u00adene bis\u00ad(phenyl carbonate) crystals was not consistent across multiple recrystallization experiments, suggesting that select impurities and/or longer, delayed evaporation methods that favor non-equilibrium products may be needed to obtain this material.Equimolar amounts of 3-amino\u00adphenol and phenyl iso\u00adcyanate were added to benzene under nitro\u00adgen and stirred for 24\u2005h. A white precipitate identified as 1-(Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017016772/su5407sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017016772/su5407Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017016772/su5407Isup3.cmlSupporting information file. DOI: 1586885CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Intra\u00admolecular amine-N\u2014H\u22efN(imine) and hy\u00addroxy-O\u2014H\u22efO(meth\u00adoxy) hydrogen bonds are noted. In the mol\u00adecular packing, amide-N\u2014H\u22efO(amide), hydroxyl-O\u2014H\u22efN(imine) and phenyl\u00adamine-N\u2014H\u22efO(meth\u00adoxy) hydrogen bonding leads to layers in the A and B) comprise the asymmetric unit of the title compound, C18H21N3O3. The urea moiety is disubstituted with one amine being linked to a phenyl ring, which is twisted out of the plane of the CN2O urea core [dihedral angles = 25.57\u2005(11) (A) and 29.13\u2005(10)\u00b0 (B)]. The second amine is connected to an imine (E conformation), which is linked in turn to an ethane bridge that links a disubstituted benzene ring. Intra\u00admolecular amine-N\u2014H\u22efN(imine) and hydroxyl-O\u2014H\u22efO(meth\u00adoxy) hydrogen bonds close S(5) loops in each case. The mol\u00adecules have twisted conformations with the dihedral angles between the outer rings being 38.64\u2005(81) (A) and 48.55\u2005(7)\u00b0 (B). In the crystal, amide-N\u2014H\u22efO(amide) hydrogen bonds link the mol\u00adecules A and B via an eight-membered {\u22efHNCO}2 synthon. Further associations between mol\u00adecules, leading to supra\u00admolecular layers in the ac plane, are hydrogen bonds of the type hydroxyl-O\u2014H\u22efN(imine) and phenyl\u00adamine-N\u2014H\u22efO(meth\u00adoxy). Connections between layers, leading to a three-dimensional architecture, comprise benzene-C\u2014H\u22efO(hy\u00addroxy) inter\u00adactions. A detailed analysis of the calculated Hirshfeld surfaces shows mol\u00adecules A and B participate in very similar inter\u00admolecular inter\u00adactions and that any variations relate to conformational differences between the mol\u00adecules.Two independent mol\u00adecules ( They have attracted considerable attention due to their wide spectrum of bio\u00adlogical activities, including anti-convulsant B]. The phenyl ring is inclined to this plane, forming a dihedral angle of 25.57\u2005(11)\u00b0 [29.13\u2005(10)\u00b0 for mol\u00adecule B]. Intra\u00admolecular N\u2014H\u22efN hydrogen bonds are found within the urea residues, Table\u00a01B]. As a result, the mol\u00adecule is twisted with the terminal rings inclined to each other, forming a (C2\u2013C7)/(C11\u2013C16) dihedral angle of 38.64\u2005(8)\u00b0 [(C20\u2013C25)/(C29\u2013C34) = 48.55\u2005(7)\u00b0 for B]. The latter represents the major difference between mol\u00adecules A and B, as illustrated in the overlay diagram shown in Fig.\u00a02E in each mol\u00adecule. Finally, each of the meth\u00adoxy substituents is twisted out of the plane of the ring to which it is bonded [C18\u2014O2\u2014C13\u2014C12 = 11.7\u2005(3) and C36\u2014O5\u2014C31\u2014C30 = \u221216.5\u2005(3)\u00b0].Two independent mol\u00adecules, a. The two mol\u00adecules comprising the asymmetric unit associate via an eight-membered amide synthon, {\u22efOCNH}2. The hy\u00addroxy-O\u2014H groups at each end of the dimeric aggregate hydrogen bond to an imine-N atom of the other independent mol\u00adecule. The hydroxyl-O3\u2014H\u22efN6(imine) inter\u00adaction is incorporated within a 10-membered {\u22efHOC2O\u22efHNCNN} heterosynthon owing to the formation of a relatively weak phenyl\u00adamine-N4\u2014H\u22efO2(meth\u00adoxy) hydrogen bond. The putative phenyl\u00adamine-N1\u2014H\u22efO5(meth\u00adoxy) hydrogen bond is beyond the standard limits . The most obvious connections between the supra\u00admolecular layers are of the type benzene-C\u2014H\u22efO(hydrox\u00adyl), which occur between centrosymmetrically related O6-benzene rings. A view of the unit-cell contents highlighting the stacking of layers is shown in Fig.\u00a03c. Other C\u2014H\u22efO and several C\u2014H\u22ef\u03c0 inter\u00adactions occur in the crystal but within the layers stabilized by hydrogen bonding. These and other weak inter\u00adactions are discussed in more detail in Analysis of the Hirshfeld surface i.e. mol\u00adecules A and B, and for overall (I)et al., 2017The Hirshfeld surface was calculated for the individual O1- and O4-mol\u00adecules in (I)O and H6O, and imine-N3 and N6 atoms on the Hirshfeld surfaces mapped over dnorm shown with labels \u20181\u2019 and \u20182\u2019 in Fig.\u00a04N and H5N, and amide-O1 and O4 atoms, i.e. \u20183\u2019 and \u20184\u2019 in Fig.\u00a04dnorm-mapped Hirshfeld surfaces with labels \u20185\u20137\u2019 in Fig.\u00a04The bright-red spots appearing near the hydroxyl-H3a and C24 in Fig.\u00a04b, of the independent mol\u00adecules, respectively, reflect short inter\u00adatomic edge-to-edge C\u22efC contacts, Table\u00a02i.e. 0.1%, to the Hirshfeld surface owing to the absence of \u03c0\u2013\u03c0 stacking between aromatic rings in the crystal, Table\u00a03A, H18A, C28, C6, C33 and C24 atoms in the images of Fig.\u00a04A and H18B atoms, Fig.\u00a04a, and O5, C35, H22 and H36C atoms, Fig.\u00a04b, indicate the contributions from the additional short inter\u00adatomic C\u22efH/H\u22efC and O\u22efH/H\u22efO contacts, Table\u00a02The presence of diminutive red spots viewed near phenyl atoms C6 in Fig.\u00a04On the Hirshfeld surfaces mapped over the electrostatic potential for the independent mol\u00adecules of (I)et al., 2007i.e. the twisting of the meth\u00adoxy substituents on the respective benzene rings and the inclination of these benzene rings with respect to the ethane bridges.It is clear from the overall two-dimensional fingerprint plots for each independent mol\u00adecule and for the entire asymmetric unit of (I)A and B have almost the same percentage contribution to their respective Hirshfeld surfaces, Table\u00a03A and B. The single short peaks at de + di \u223c 2.1\u2005\u00c5 in the delineated plots for both the mol\u00adecules indicate the involvement of hydrogen atoms of both in short inter\u00adatomic H\u22efH contacts, Table\u00a02de + di \u223c 2.0\u2005\u00c5 (inner region) and at \u223c 2.2\u2005\u00c5 (outer region) in the fingerprint plots delineated into O\u22efH/H\u22efO and N\u22efH/H\u22efN contacts, respectively. The forceps-like distribution of points linked with the donor spike for mol\u00adecule A and the acceptor spike for mol\u00adecule B at de + di \u223c 2.5\u2005\u00c5 in the fingerprint plots delineated into O\u22efH/H\u22efO contacts are due to weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions and the short inter\u00adatomic contacts summarized in Table\u00a02de + di \u223c 2.6\u2005\u00c5 in the acceptor and donor regions of fingerprint plots delineated into C\u22efH/H\u22efC contacts for mol\u00adecules A and B, respectively, represent the involvement of these atoms in the short inter\u00adatomic C\u22efH/H\u22efC contacts, Table\u00a02de + di \u223c 2.7\u2005\u00c5 in the donor and acceptor regions of mol\u00adecules A and B, respectively. The other C\u22efO/O\u22efC, O\u22efO and C\u22efC inter\u00adatomic contacts summarized in Table\u00a03The fingerprint plot delineated into H\u22efH contacts for mol\u00adecules et al., 2016et al., 1980et al., 1980et al., 2011et al., 2014et al., 2014There are no direct precedents for the structure of (I)in vacuo; yield: 75%. Light-yellow prisms of (I)v/v 20\u2005ml). IR (cm\u22121): 3201 \u03bd(N\u2014H), 1670 \u03bd(C=N), 1213 \u03bd(C\u2014N), 1026 \u03bd(C=O). MS m/z: 327.25 [M+1]+Analytical grade reagents were used as procured without further purification. 4-Phenyl\u00adsemicarbazide and vanillylacetone were dissolved sep\u00adarately in hot absolute ethanol (30\u2005ml) and mixed with stirring. The reaction mixture was heated and stirred for 20\u2005min., then stirred for another 30\u2005min. at room temperature. The resulting white precipitate was filtered off, washed with cold absolute ethanol and dried Uiso(H) set to 1.2\u20131.5Ueq(C). The oxygen- and nitro\u00adgen-bound H atoms were located in a difference-Fourier map but were refined with distance restraints of O\u2014H = 0.84\u00b10.01\u2005\u00c5 and N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.5Ueq(O) and 1.2Ueq(N), respectively. The maximum and minimum residual electron density peaks of 0.60 and 0.26\u2005e\u2005\u00c5\u22123, respectively, were located 0.95 and 0.75\u2005\u00c5 from atoms H10A and H36A, respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989017017273/hb7720sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989017017273/hb7720Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017017273/hb7720Isup3.cmlSupporting information file. DOI: 926756CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports6: Article number: 3657910.1038/srep36579; published online: 11152016; updated: 05042017This Article contains errors. In the legend of Figure 1,\u22121\u201d\u201c1.25\u2009cGy\u2009minshould read:\u22121\u201d.\u201c1.25\u2009Gy\u2009minIn addition, in the Methods section under subheading \u2018Animals and Radiation Exposure\u2019,\u22121\u201d\u201cFor serial tissue analysis, age and sex-matched C57BL/6J and C57L/J mice at 10\u201312 weeks of age were irradiated to the whole lung with a single dose of 15\u2009Gy of 320\u2009kVp X-rays at a dose rate of 1.25\u2009cGy\u2009minshould read:\u22121\u201d\u201cFor serial tissue analysis, age and sex-matched C57BL/6J and C57L/J mice at 10\u201312 weeks of age were irradiated to the whole lung with a single dose of 15\u2009Gy of 320\u2009kVp X-rays at a dose rate of 1.25\u2009Gy\u2009min"} +{"text": "In the crystal, N+(C)\u2014H\u22efCl\u2212 hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions lead to the formation of a three-dimensional supra\u00admolecular network.The title salt, C 18H26N42+\u00b72Cl\u2212, was obtained by the protonation of N,N-bis\u00ad(pyridin-4-ylmeth\u00adyl)cyclo\u00adhexane-1,4-di\u00adamine with hydro\u00adchloric acid in ethanol. The asymmetric unit consists of one half of an N,N-bis\u00ad(pyridin-3-ylmeth\u00adyl)cyclo\u00adhexane-1,4-di\u00adammonium dication, with a crystallographic inversion centre located at the centre of the cyclo\u00adhexyl ring, and a chloride anion. The central cyclo\u00adhexyl ring in the dication adopts a chair conformation. The two trans-(4-pyridine)\u2013CH2\u2013NH2\u2013 moieties at the 1- and 4-positions of the central cyclo\u00adhexyl ring occupy equatorial sites. The terminal pyridine ring is tilted by 53.72\u2005(6)\u00b0 with respect to the mean plane of the central cyclo\u00adhexyl ring (r.m.s. deviation = 0.2413\u2005\u00c5). In the crystal, N+\u2014H\u22efCl\u2212 hydrogen bonds between the dications and the chloride anions, and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings of the dications afford a two-dimensional sheet extending parallel to the ab plane. These sheets are further connected through weak C\u2014H\u22efCl\u2212 hydrogen bonds, resulting in the formation of a three-dimensional supra\u00admolecular network.The title salt, C In the dication, the central cyclo\u00adhexyl ring displays a chair conformation and the two trans-(4-pyridine)\u2013CH2\u2013NH2\u2013 moieties occupy equatorial sites at the 1- and 4-positions of the central cyclo\u00adhexyl ring. The terminal pyridine ring is tilted by 53.72\u2005(6)\u00b0 with respect to the mean plane of the cyclo\u00adhexyl ring (r.m.s. deviation = 0.2413\u2005\u00c5). This tilting angle is larger than that [27.98\u2005(5)\u00b0] of the similar dication with 4-pyridine rings as the terminal groups \u2005\u00c5] between the pyridine rings cyclo\u00adhexane-1,4-diaminium)cobalt(II) penta\u00adnitrate methanol solvate] cyclo\u00adhexane-1,4-di\u00adamine, prepared according to a literature method = 0.95\u2005\u00c5 for Csp2\u2014H, 0.99\u2005\u00c5 for methyl\u00adene, 1.00\u2005\u00c5 for methine H atoms] and were refined as riding with Uiso(H) = 1.2Ueq(C). The N-bound H atoms involved in hydrogen bonds were located in difference Fourier maps and refined freely [N\u2014H = 0.891\u2005(15) and 0.876\u2005(16)\u2005\u00c5].Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989016017205/hg5479sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989016017205/hg5479Isup2.hklStructure factors: contains datablock(s) I. DOI: 1511616CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "ATP\u2010sensitive P2X7 receptor activates the NLRP3 inflammasome and is a key component of the innate immune system. We used a fluid\u2010resuscitated rat model of fecal peritonitis and acute kidney injury (AKI) to investigate the contribution of this purinergic receptor to renal dysfunction in sepsis. Six and 24\u00a0h time\u2010points were chosen to represent early and established sepsis, respectively. A selective P2X7 receptor antagonist (A\u2010438079) dissolved in dimethyl sulfoxide (DMSO) was infused 2\u00a0h following induction of sepsis. Compared with sham\u2010operated animals, septic animals had significant increases in heart rate (\u22121(\u22124 to 8)% vs. 21(12\u201326)%; P\u00a0=\u00a00.003), fever (37.4(37.2\u201337.6)\u00b0C vs. 38.6(38.2\u201339.0)\u00b0C; P\u00a0=\u00a00.0009), and falls in serum albumin (29(27\u201330)g/L vs. 26(24\u201328); P\u00a0=\u00a00.0242). Serum IL\u20101\u03b2 (0(0\u201310)(pg/mL) vs. 1671(1445\u201333778)(pg/mL); P\u00a0<\u00a00.001) and renal IL\u20101\u03b2 (86(50\u2013102)pg/mg protein vs. 200 (147\u2013248)pg/mg protein; P\u00a0=\u00a00.0031) were significantly elevated in septic compared with sham\u2010operated animals at 6\u00a0h. Serum creatinine was elevated in septic animals compared with sham\u2010operated animals at 24\u00a0h (23(22\u201325) \u03bcmol/L vs. 28 (25\u201330)\u03bcmol/L; P\u00a0=\u00a00.0321). Renal IL\u20101\u03b2 levels were significantly lower in A\u2010438079\u2010treated animals compared with untreated animals at 6\u00a0h (70(55\u2013128)pg/mg protein vs. 200(147\u2013248)pg/mg protein; P\u00a0=\u00a00.021). At 24\u00a0h, compared with untreated animals, A\u2010438079\u2010treated animals had more rapid resolution of tachycardia (22(13\u201336)% vs. \u22121(\u22126 to 7)%; P\u00a0=\u00a00.019) and fever (39.0(38.6\u201339.1)\u00b0C vs. 38.2(37.6\u201338.7)\u00b0C; P\u00a0<\u00a00.024), higher serum albumin (23(21\u201325)g/L vs. (27(25\u201328)g/L); P\u00a0=\u00a00.006), lower arterial lactate (3.2(2.5\u20134.3)mmol/L vs. 1.4(0.9\u20131.8)mmol/L; P\u00a0=\u00a00.037), and lower serum creatinine concentrations (28(25\u201330)\u03bcmol/L vs. 22(17\u201327)\u03bcmol/L; P\u00a0=\u00a00.019). P2X7A treatment ameliorates the systemic inflammatory response and renal dysfunction in this clinically relevant model of sepsis\u2010related AKI.Sepsis is a major clinical problem associated with significant organ dysfunction and high mortality. The Sepsis is a common and serious global health issue, accounting for 20% of all admissions to intensive care units (ICU), and is the leading cause of death in noncardiac ICUs mice subjected to cecal ligation and puncture have an attenuated inflammatory response and reduced lung injury compared with wild\u2010type mice , P2X7 receptor contributes to renal injury in sepsis through an inflammatory response with local production of cytokines. We aimed to define the expression of this receptor within the kidney, and its relationship with cytokine production, and renal function; we sought to assess effects of P2X7 antagonism in a rat model of sepsis\u2010induced AKI. The primary objective of this study was to evaluate the effect of a selective P2X7 antagonist (A\u2010438079) in preventing renal dysfunction (as defined by an elevated serum creatinine) in an experimental model of sepsis.We hypothesized that upregulation of the ATP\u2010sensitive P2XAll animal experiments were performed under a Home Office Project License (PPL 70/7029) and local UCL Ethics Committee approval.7 receptor antagonist (A\u2010438079) dissolved in DMSO. Details on monocyte isolation from male Wistar rats are detailed in the supplementary data. Monocytes require priming with LPS prior to release of IL\u20101\u03b2 by co\u2010stimulation with ATP . The duration of LPS\u2010priming is variable, although 4\u00a0h has been shown to be adequate was kindly donated by Abbvie . Based on previously published data and personal communication with Abbvie, the pharmacokinetic profile of A\u2010438079 was determined. A\u2010438079 blocked BzATP (10\u00a0\u03bcmol/L)\u2013mediated changes in intracellular Ca++ concentrations at rat P2X7 receptors but not at other P2 receptors would achieve therapeutic levels in serum. We therefore administered a dose of 10\u00a0\u03bcg/kg. The antagonist was given as a single bolus to achieve a peak (therapeutic) concentration, followed by 4\u00d7 the dose for the first half\u2010life, followed by 2\u00d7 the dose for the second half life, followed by 10\u00a0\u03bcg/(kg\u00b7h) thereafter.Unpublished data from Abbvie Pharmaceuticals demonstrates that the half\u2010life of intravenous A\u2010438079 is 0.69\u00a0h (41\u00a0min). An intravenous dose of 10\u00a0\u03bcmol/L, the pH at room temperature fell to 2.2, and at 38\u00b0C was 1.6. Dimethylsulfoxide (DMSO) was therefore selected as a drug solvent. Due to its amphipathic properties, DMSO is an effective solvent for water\u2010insoluble compounds and is a hydrogen\u2010bound disrupter. As such it is a commonly used solvent for many drugs in pharmacological studies. This is a commonly used drug solvent, however, itself has biological activity weighing 300\u2013375\u00a0g were used throughout. All experiments were performed in accordance with relevant guidelines and regulations and are detailed in the supplementary data and our characterization manuscript initially comprised sham\u2010operated and untreated septic animals at both 6\u00a0h and 24\u00a0h time points, and DMSO\u00a0\u00b1\u00a0A\u2010438079 treatment at 6\u00a0h and 24\u00a0h. On review of the results, we undertook further studies where twice the dose of A\u2010438079 was administered.Experimental groups .DuoSet ELISA kits were used according to the manufacturer's instructions. The antibody supplied in the IL\u20101\u03b2 in sham and septic animals at 24\u00a0h. Details of methods for Western blot are included in the supplementary data.Protein was extracted and estimated from whole kidney tissue. Four animals were randomly selected from each of the sham and sepsis groups, for analysis. Kidney homogenate were assessed for expression of Caspase\u20101, and IL\u20101The number of antagonist\u2010treated animals and placebo\u2010treated controls was calculated based on previous laboratory experience. We regarded a 20% change in a tested variable as an important biological effect. The study was designed to have a power of 90% and a significance level of 5%. At least 6 replicate animals per group per time point.U test was used for comparison of continuous variables between 2 groups. A P\u00a0<\u00a00.05 was taken as statistically significant.Graphpad Prism was used for statistical analyses and graphs. All data were regarded as nonparametric due to small sample sizes (6\u201312 per group). Continuous variables are presented as median (interquartile range). Data were analyzed in prospectively defined groups. All 6\u00a0h experiments were analyzed as a predefined group, as were all 24\u00a0h experiments. For comparison of continuous variables between more than two groups, Kruskal\u2013Wallis test with post hoc Dunn's test is used. Mann\u2013Whitney \u03b2 production (P\u00a0=\u00a00.029). Addition of 2% DMSO significantly inhibited the release of IL\u20101\u03b2 (P\u00a0=\u00a00.025). Addition of 10\u00a0\u03bcmol/L A\u2010438079 had no added effect on the reduction in IL\u20101\u03b2 release. 5\u00a0\u03bcmol/L brilliant blue G (BBG) had a significant inhibitory effect on monocyte IL\u20101\u03b2 release (P\u00a0=\u00a00.029) stimulation of cultured monocytes resulted in significant IL\u20101P\u00a0=\u00a00.003), fever (37.4(37.2\u201337.6)\u00b0C vs. 38.6(38.2\u201339.0)\u00b0C; P\u00a0=\u00a00.0009), and falls in serum albumin (29(27\u201330)g/L vs. 26(24\u201328); P\u00a0=\u00a00.0242). Serum IL\u20101\u03b2 (0(0\u201310)(pg/mL) vs. 1671(1445\u201333778)(pg/mL); P\u00a0<\u00a00.001) and renal IL\u20101\u03b2 (86(50\u2013102)pg/mg protein vs. 200 (147\u2013248)pg/mg protein; P\u00a0=\u00a00.0031) were significantly elevated in septic compared with sham\u2010operated animals at 6\u00a0h. Serum creatinine, however, remained unchanged at 6\u00a0h (25(23\u201327)\u03bcmol/L vs. 23(21\u201328) \u03bcmol/L; P\u00a0=\u00a00.9155) in sham\u2010operated compared with septic animals.Clinical, biochemical, and cytokine changes in the septic model are detailed in our characterization manuscript , fever (37.5(37.4\u201337.8)\u00b0C vs. 39.0(38.6\u201339.1)\u00b0C; P\u00a0=\u00a00.0002), and fall in serum albumin (30(29\u201331)g/L vs. 23(21\u201325)g/L; P\u00a0=\u00a00.0003). Serum IL\u20101\u03b2 (0(0\u201310)(pg/mL) vs. 1463(927\u20132541)(pg/mL); P\u00a0<\u00a00.001) remains significantly elevated in septic compared with sham\u2010operated animals at 24\u00a0h although renal IL\u20101 \u03b2 levels were similar (168(154\u2013176)pg/mg protein vs. 186(149\u2013280)pg/mg protein; P\u00a0=\u00a00.053). Serum creatinine was significantly elevated in septic animals compared with sham\u2010operated animals at 24\u00a0h (23(22\u201325)\u03bcmol/L vs. 28 (25\u201330)\u03bcmol/L; P\u00a0=\u00a00.0321).At 24\u00a0h, septic animals had persistent changes of systemic inflammation Table\u00a0, Fig.\u00a02.7 on immunohistochemistry: there was minimal constitutive P2X7 expression limited to distal tubular epithelial cells protein expression in septic animals compared with sham\u2010operated animals vs. 1.35(1.25\u20131.37); \u03bcmol/L did not result in any clinical or biochemical differences between untreated and A\u2010438079\u2010treated septic animals (data not shown). We therefore undertook further studies where twice the dose of A\u2010438079 was administered.Initial in\u00a0vivo experiments targeting a peak concentration of 3\u00a0\u03b2 and between treated and untreated animals pg/mg protein vs. 200(147\u2013248)pg/mg protein; P\u00a0=\u00a00.021). Arterial lactate concentration was different between DMSO\u2010treated and untreated animals (1.0 (0.9\u20131.0)mmol/L vs. 1.2(1.0\u20131.3)mmol/L; P\u00a0=\u00a00.033), although remained within the normal clinical range in all groups of animals.At 6\u00a0h there was no difference among the groups in core temperature, change in heart rate, serum creatinine, or serum IL\u20101ls Table\u00a0. Renal IP\u00a0=\u00a00.019) and fever (39.0(38.6\u201339.1)\u00b0C vs. 38.2(37.6\u201338.7)\u00b0C; P\u00a0<\u00a00.024), higher serum albumin (23(21\u201325)g/L vs. (27(25\u201328)g/L); P\u00a0=\u00a00.006), lower arterial lactate (3.2(2.5\u20134.3)mmol/L vs. 1.4(0.9\u20131.8)mmol/L; P\u00a0=\u00a00.037), and lower serum creatinine concentrations (28(25\u201330)\u03bcmol/L vs. 22(17\u201327)\u03bcmol/L; P\u00a0=\u00a00.019).At 24\u00a0h, there were a number of significant differences between A\u2010438079 treated animals and untreated septic animals Table\u00a0, Fig.\u00a02.P\u00a0=\u00a00.016), and lower arterial lactate (2.2 (2.1\u20132.2)mmol/L vs. 1.4(0.9\u20131.8)mmol/L; P\u00a0=\u00a00.001). Serum IL\u20101\u03b2 and renal IL\u20101\u03b2 were similar in A\u2010438079\u2010treated, DMSO\u2010treated, and untreated animals at 24\u00a0h.At 24\u00a0h, compared with DMSO\u2010treated animals, A\u2010438079\u2010treated animals had higher serum albumin (24(22\u201325)g/L vs. 27(25\u201328)g/L; 7 and IL\u20101\u03b2, are expressed de novo within the renal tubular epithelial cells in this septic model. This is consistent with published data showing that LPS\u2010primed Madin\u2013Darby canine kidney (MDCK) renal tubular epithelial cells express Toll\u2010like receptor 4, NLRP3, caspase\u20101, and IL\u20101\u03b2 mRNA , though P2X7 activation accelerates mature IL\u20101\u03b2 release from monocytes levels of IL\u20101\u03b2 \u2010producing Escherichia coli had markedly lower survival, higher cytokine levels (including IL\u20101\u03b2), and activated intravascular coagulation compared with wildtype mice transplanted with wild\u2010type kidneys were protected from LPS\u2010induced AKI, whereas wild\u2010type mice transplanted with C3H/HeJ kidneys developed severe LPS\u2010induced AKI. This suggests that TLR4 expression in circulating cells propagates injury in septic AKI, rather than intra\u2010renal TLR4 , with better perfusion to organs including the kidney, rather than specific actions over the renal\u2010inflammasome activation.The changes found in animals treated with the P2X\u03b2 measurement, which does not allow localization to specific cell types. Immunohistochemistry would enable localization of these cytokines. Although insights into renal expression of the inflammasome and P2X7 in sepsis have been demonstrated, the expression of the P2X7 in other organs and on immune cells also needs to be evaluated.ELISAs were performed on whole kidney homogenate for IL\u2010\u03b2 and tubular cell P2X7 expression by 6\u00a0h and 24\u00a0h, respectively. Treatment with A\u2010438079 after the onset of sepsis abrogated the rise in serum creatinine at 24\u00a0h with reduced renal IL\u20101\u03b2 expression early. In addition, A\u2010438079 treatment reduced fever, improved resolution of tachycardia, increased serum albumin concentration and reduced lactate at 24\u00a0h. Given these encouraging results in an in\u00a0vivo model of sepsis that is relevant to human disease, further studies targeting P2X7 in sepsis are warranted.We demonstrate a number of strengths of our experimental model relevant to potential therapeutics in sepsis. Sepsis is associated with an increase in renal IL\u20101All animal experiments were performed under a Home Office Project License (PPL 70/7029) and local University College London Ethics Committee approval.7 antagonist (A\u2010438079). FWKT has received research project grants from AstraZeneca Limited, Baxter Biosciences, Boehringer Ingelheim, and MedImmune. He has consultancy agreements with Rigel Pharmaceuticals, Novartis and Baxter Biosciences.Materials: Abbvie pharmaceuticals have provided the selective P2XData S1. Supplementary data.Click here for additional data file."} +{"text": "Scientific Reports6: Article number: 18631; 10.1038/srep18631 published online: 01112016; updated: 09012016.In this Article and Supplementary Information file, some instances of \u20180.001M PBS\u2019 and \u20180.01M PBS\u2019 are incorrectly written as \u20180.01M PBS\u2019 and \u20180.1M PBS\u2019 respectively. With the exception of the following:In the Materials and Methods section, under subheading \u2018Specimen Preparation\u2019,\u201cFixed samples were incubated in A4P0 hydrogel monomer solution (4% acrylamide in 0.1\u2009M PBS) supplemented with 0.25% of the photoinitiator 2,2\u2032-azobis[2-(2-imidazolin-2-yl)propane] dihydrochloride overnight at 4\u2009\u00b0C\u201d.should read:\u201cFixed samples were incubated in A4P0 hydrogel monomer solution (4% acrylamide in 0.001\u2009M PBS) supplemented with 0.25% of the photoinitiator 2,2\u2032-azobis[2-(2-imidazolin-2-yl)propane] dihydrochloride overnight at 4\u2009\u00b0C\u201d.Under subheading \u2018ACT Reagents\u2019,A4P0 solution, Forty ml of 40% acrylamide was added to 360\u2009ml dH2O to prepare 400\u2009ml of the hydrogel monomer solution (A4P0)\u201d.\u201cshould read:A4P0 solution, Forty ml of 40% acrylamide was added to 360\u2009ml 0.001M PBS to prepare 400\u2009ml of the hydrogel monomer solution (A4P0)\u201d.\u201cUnder subheading \u2018Whole-body and Organ ACT Protocol,\u201cHigher sodium concentrations in the clearing solution (0.03\u2009M and 0.1\u2009M PBS) ameliorated tissue swelling but resulted in tissue thinning (data not shown)\u201d.should read:\u201cHigher sodium concentrations in the clearing solution (0.003\u2009M and 0.01\u2009M PBS) ameliorated tissue swelling but resulted in tissue thinning (data not shown)\u201d.\u201cThe head was freed from subcutaneous skin and hair and washed several times with 0.1\u2009M PBS.should read:\u201cThe head was freed from subcutaneous skin and hair and washed several times with 0.001\u2009M PBS\u201d.In the Supplementary Information file, under subheading \u2018\u2019Hydrogel monomer infusion & polymerization,\u201c1. Prepare the A4P0 hydrogel monomer solution.Take 40\u2009ml of 4% acrylamide in 0.1\u2009M PBS solution and add 100\u2009mg of VA-044 Initiator to make a final concentration of 0.25%\u201d.should read:\u201c1. Prepare the A4P0 hydrogel monomer solution.Take 40\u2009ml of 4% acrylamide in 0.001\u2009M PBS solution and add 100\u2009mg of VA-044 Initiator to make a final concentration of 0.25%\u201d.Under the subheading \u2018Immunolabeling ACT-processed brain tissue\u2019,\u201c1. Wash samples in 0.1\u2009M PBS for 3\u20135\u2009hours and change the buffer every hour\u201d.should read:\u201c1. Wash samples in 0.001\u2009M PBS for 3\u20135\u2009hours and change the buffer every hour\u201d.\u201c3. Wash samples in 0.1\u2009M PBS for 3\u20135\u2009hours, and change buffer every hour\u201d.should read:\u201c3. Wash samples in 0.001\u2009M PBS for 3\u20135\u2009hours, and change buffer every hour\u201d."} +{"text": "The title salt forms a racemate due to disorder of the hy\u00addroxy group [occupancy ratio 0.738\u2005(3):0.262\u2005(3)] at the stereogenic C atom. 13H22NO3+\u00b7C7H5O2\u2212, comprises one salbutamol cation {sys\u00adtematic name: 4-[2-(tert-butyl\u00adaza\u00adnium\u00adyl)-1-hy\u00addroxy\u00adeth\u00adyl]-2-(hy\u00addroxy\u00admeth\u00adyl)phenol} and a benzoate anion. The cation shows disorder of the hy\u00addroxy group [occupancy ratio 0.738\u2005(3):0.262\u2005(3)] at the stereogenic C atom. The non-planar benzoate anion [the dihedral angle between the benzene ring and the carboxyl group is 11.30\u2005(8)\u00b0] is linked to the salbutamol cation by a medium-strength O\u2014H\u22efO hydrogen bond. Other inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds of weaker nature give rise to [001] chains.The title salt, C The bond lengths of the carboxyl\u00adate group of the BA\u2212 anion, C20\u2014O4 and C20\u2014O5, are 1.2617\u2005(15) and 1.2604\u2005(15)\u2005\u00c5, respectively. The slight difference may be caused by the role of O4 as an acceptor atom of the O3\u2014H3\u22efO4 hydrogen bond with one of the hy\u00addroxy groups of +SAL. The +SAL cation also has an intra\u00admolecular hydrogen bond between the two hy\u00addroxy functions (O1\u2014H1\u22efO3), forming an S(6) ring motif \u00b0. There is some disorder at the stereogenic centre (C8) of the +SAL cation, but the space group is centrosymmetric and the +SAL cation is racemic.The +SAL cation is connected to the \u2212BA anion via a medium-strength O3\u2014H3\u22efO4 hydrogen bond was determined and benzoic acid were added to 10\u2005ml methanol and stirred for 3\u2005h. The solvent was then evaporated at room temperature to yield salbutamol benzoate. After recrystallization from water, pure crystals were again dissolved in ethanol and the solution filtered. The neat filtrate was evaporated slowly to give colourless block-like single crystals of salbutamol benzoate.Uiso(H) = 1.5Ueq(C) for methyl H atoms and Uiso(H)\u00a0= 1.2Ueq(C) for all other H atoms. The H atoms of the NH2 group and the hy\u00addroxy group were also constrained to ideal values and allowed to ride in the refinement, with Uiso(H) = 1.2Ueq(N) and 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989017011513/wm5403sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989017011513/wm5403Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989017011513/wm5403Isup3.cmlSupporting information file. DOI: 1482124CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The chemical modifications in terms of changes in substituents in the title compounds have not affected the type nor strength of two defining inter\u00admolecular inter\u00adactions present in both crystal structures. 28H26N2O2, (I), and C27H22BrClN2O2, (II), differ in their substituents, viz.4-methyl\u00adphenyl and benzoyl rings in (I) replaced by 2-chloro\u00adphenyl and 4- bromo\u00adbenzoyl, respectively, in (II). A significant difference between the two mol\u00adecules is found in the deviation of the benzoyl O atom from the least-squares plane of the ring to which it is attached , a fact which may be attributed to the different participation of the benzoyl O atoms as acceptors in their inter\u00admolecular C\u2014H\u22efO inter\u00adactions. The chemical modifications in (I) and (II) do not seem to affect the type nor strength of the inter\u00admolecular N\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds responsible for the two crystal structures, such that the aggregation of mol\u00adecules appears similar in spite of the mol\u00adecular changes.The two title compounds, C Indole, a pharmacologically significant nucleus, is known for anti-inflammatory viz. N2\u2013C9\u2013C10\u2013C11\u2013C12 and N2\u2013C2\u2013C14\u2013C13\u2013C12 are, respectively, Q = 0.362\u2005(4)\u2005\u00c5, \u03c6 = 264.4\u2005(5)\u00b0 indicating a twist about C10\u2013C11, and Q = 0.408\u2005(3)\u2005\u00c5, \u03c6=67.9\u2005(4)\u00b0 conforming to an envelope on C14. The corresponding values in (II)Q = 0.378\u2005(3)\u2005\u00c5, \u03c6 = 82.4\u2005(4)\u00b0 and Q = 0.423\u2005(3)\u2005\u00c5, \u03c6 = 251.4\u2005(3)\u00b0, may differ slightly from those in (I)Q of the fused eight-membered pyrrolizine and the nine-membered indolone ring systems are respectively, 0.727\u2005(3) and 0.129\u2005(3)\u2005\u00c5 in (I)Mol\u00adecular diagrams of (I)In both compounds, the spiro-fused ring systems tend to be rigid by remaining nearly perpendicular to each other, whereas the remaining substituted rings appear to be more \u2018compromising\u2019 towards hydrogen-bonding requirements, irrespective of their intra- or inter\u00admolecular nature. As an example, the free rotation of the benzoyl group in (II)A significant difference between the two structures is observed in the deviation of benzoyl atom O2 from the least-squares plane of the C15\u2013C21 atoms: 0.593\u2005(4) in (I)etc, these mol\u00adecular modifications do not seem to affect the type nor strength of the two relevant N\u2014H\u22efN and C\u2014H\u22efO inter\u00admolecular hydrogen bonds defining the crystal structures (Tables 1x\u00a0+\u00a0y\u00a0\u2212\u00a0z\u00a0+\u00a0dO\u22efBr= 3.192\u2005(2)\u2005\u00c5] is present in structure (II)Even if the differences in the substituents produce differences in lattice types, space group, cell metrics, Tables 1 and 2 \u25b8,et al., 2016et al., 1998A search of the Cambridge Structural Database -1-phenyl-3-(p-tol\u00adyl)prop-2-en-1-one (0.4\u2005mmol) [for the synthesis of (II)E)-1-(4-bromo\u00adphen\u00adyl)-3-(2-chloro\u00adphen\u00adyl)prop-2-en-1-one (0.4\u2005mmol)], isatin (0.4\u2005mmol) and l-proline (0.4\u2005mmol), which was dissolved in 5\u2005ml of methanol, and 1\u2005mol% of CMPTC was added and stirred at reflux temperature until the completion of reaction as indicated by TLC. After this step, the mixture was poured onto ice; the precipitate was filtered and recrystallized from ethanol solution, to get the pure product without column chromatography.The synthesis of (I)Uiso(H) set at 1.2\u20131.5Ueq(C). Compound (I)Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989016016741/bg2594sup1.cifCrystal structure: contains datablock(s) I, II, c1. DOI: 1503430, 1503429CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The ethyl\u00adeneglycol ligand forms a non-planar metallacyclic ring by chelating the Ru atom via the O atoms. The O\u22efO distances of 2.554\u2005(2) and 2.568\u2005(2)\u2005\u00c5 are indicative of hydrogen bonding between coordinated ethyl\u00adeneglycol and outer-sphere trifluoro\u00admethane\u00adsulfonate fragments. The crystal packing is stabilized by ionic forces and several CH3\u22ef\u00b7F (2.585 and 2.640\u2005\u00c5) and CH3\u22efO inter\u00adactions between the penta\u00admethyl\u00adcyclo\u00adpenta\u00addienyl ligand and trifluoro\u00admethane\u00adsulfonate anion. There is noticeable short inter\u00admolecular contact [2.9039\u2005(16)\u2005\u00c5], between an O atom of the SO3 group and a C atom of the penta\u00admethyl\u00adcyclo\u00adpenta\u00addienyl ligand.The title compound, [Ru(C DOI: 10.1107/S1600536807067426/bg2160Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Cu2(C8H7O2)4(C9H7N)2], the two Cu cations are bridged by four carboxyl\u00adate groups of the phenyl\u00adacetate anions; each Cu cation is further coordinated by an isoquinoline ligand to complete the distorted CuO4N square-pyramidal geometry. The Cu cation is displaced by 0.2092\u2005(8)\u2005\u00c5 from the basal plane formed by the four O atoms. Within the dinuclear mol\u00adecule, the Cu\u22efCu separation is 2.6453\u2005(6)\u2005\u00c5. Although a parallel, overlapped arrangement of isoquinoline ligands exists in the crystal structure; the longer face-to-face distance of 3.667\u2005(5)\u2005\u00c5 suggests there is no \u03c0\u2013\u03c0 stacking between isoquinoline ring systems.In the title centrosymmetric binuclear Cu DOI: 10.1107/S1600536809048697/ng2685Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the cation, the FeCo2 triangle is symmetrically capped on one face by an S atom and on the other by a C atom linked to a methyl trithio\u00adcarbonate residue that bridges the Fe\u2014C bond. Each Co atom carries a cyclo\u00adpenta\u00addienyl ligand while the Fe atom coordinates to one carbonyl and one triphenyl\u00adphosphine ligand. In the crystal structure, the cation is linked to the anion by a number of weak non-classical C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds and weak S\u22efO (3.317\u2005\u00c5) and S\u22efF (3.198\u2005\u00c5) inter\u00adactions. The structure is further stabilized by additional inter\u00admolecular C\u2014H\u22efO, C\u2014H\u22efF and O\u22efO (2.942\u2005\u00c5) contacts, together with an unusual S\u22ef\u03c0(Cp) inter\u00adaction (S\u22efcentroid distance = 3.385\u2005\u00c5), generating an extended network.The asymmetric unit of the title compound, [FeCo DOI: 10.1107/S1600536808008970/hb2713Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The initial reactant of CuI was oxidized to CuII. The asymmetric unit contains two independent complex mol\u00adecules, two I\u2212 ions and one water molecule. Each CuII atom is coordinated by two O atoms from a 4-methyl\u00adbenzoate ligand and four N atoms from two 2,2\u2032-bipyridine ligands, displaying a distorted octa\u00adhedral geometry. The structure involves O\u2014H\u22efI hydrogen bonds between the water mol\u00adecule and iodide ions and \u03c0\u2013\u03c0 stacking inter\u00adactions between the benzene and pyridyl rings [centroid\u2013centroid distance = 3.79\u2005(1)\u2005\u00c5] and between the pyridyl rings [centroid\u2013centroid distance = 3.87\u2005(1)\u2005\u00c5].The title compound, [Cu(C DOI: 10.1107/S1600536808024252/hy2148Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The N and O donor atoms are mutually cis. The dihedral angle between two benzene rings of the ligand is 38.86\u2005(8)\u00b0. There are also three solvent water mol\u00adecules, two of which lie across different crystallographic twofold rotation axes; one of these is partially occupied with a refined occupancy factor of 0.570\u2005(7). The water mol\u00adecules are linked together as tetra\u00admers in R 2 2(8) ring motifs, which also connect two neighbouring mol\u00adecules of the complex through a network of O\u2014H\u22efO hydrogen bonds. The crystal structure is further stabilized by inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions, which link neighbouring mol\u00adecules into extended chains along the b axis. Other inter\u00adesting features of the crystal structure are the short inter\u00admolecular C\u22efC [3.204\u2005(3)\u20133.365\u2005(3)\u2005\u00c5] and the C\u22efO [3.199\u2005(2)\u20133.205\u2005(2)\u2005\u00c5] contacts which are shorter than the sum of the van der Waals radii of these atoms.In the title complex, [Ni(C DOI: 10.1107/S1600536809014500/sj2621Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, O\u2014H\u22efO hydrogen bonds between the coordinated water mol\u00adecules and uncoordinated carboxyl\u00adate O atoms, and weak \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 4.105\u2005(2)\u2005\u00c5] between the benzene and pyrazine rings lead to a three-dimensional supra\u00admolecular network.In the title compound, [Co(C DOI: 10.1107/S1600536810037906/hy2354Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "A supra\u00admolecular network is formed via inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen-bonding inter\u00adactions. \u03c0\u2013\u03c0 stacking inter\u00adactions between neighboring pyridine rings are also present, the centroid\u2014centroid distance being 3.808\u2005(2)\u2005\u00c5.In the title complex, [Cu(C DOI: 10.1107/S1600536810035555/pv2323Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Both MnIII centers are in a distorted octa\u00adhedral geometry with the N2O2 donor atoms of the tetra\u00addentate Schiff base dianion in the equatorial plane. The axial positions in the coordination environment of one MnIII complex are occupied by a chloride ion and a water mol\u00adecule, but a methanol mol\u00adecule replaces the water mol\u00adecule in the other complex. The coordinated water mol\u00adecule takes part in an O\u2014H\u22efCl hydrogen bond between the two MnIII complexes. In the crystal structure, O\u2014H\u22efCl hydrogen bonds link the mol\u00adecules into infinite one-dimensional chains along the [100] direction. The crystal structure is stabilized by O\u2014H\u22efCl hydrogen bonds together with weak C\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions. A C\u2014H\u22ef\u03c0 inter\u00adaction is also observed in the crystal structure.The asymmetric unit of the title complex, [Mn(C Cl(H2O)][Mn(C28H30N2O2)Cl(CH4O)] = 0.057 wR(F 2) = 0.147 S = 1.03 10108 reflections653 parametersH-atom parameters constrainedmax = 0.56 e \u00c5\u22123 \u0394\u03c1min = \u22120.52 e \u00c5\u22123 \u0394\u03c1 APEX2 (Bruker, 2005APEX2; data reduction: SAINT (Bruker, 2005SHELXTL (Sheldrick, 2008SHELXTL; molecular graphics: SHELXTL; software used to prepare material for publication: SHELXTL and PLATON (Spek, 2003Data collection: 10.1107/S1600536808006818/sj2472sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536808006818/sj2472Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the anion, the GaIII ion is coordinated by four O atoms [Ga\u2014O = 1.9706\u2005(16)\u20132.0494\u2005(15)\u2005\u00c5] and two N atoms [Ga\u2014N = 1.9660\u2005(18) and 1.9709\u2005(17)\u2005\u00c5] from two pydc ligands in a distorted octa\u00adhedral geometry. The crystal structure exhibits inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances of 3.5359\u2005(13) and 3.6550\u2005(14)\u2005\u00c5].The asymmetric unit of the title compound, (C DOI: 10.1107/S1600536808029140/cv2438Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The O atoms of two 4-pyridone ligands bridge two symmetrically related AgNO3 units, forming a dimer, with an Ag\u22efAg separation of 3.680\u2005(2)\u2005\u00c5. Neighbouring dimers are linked into an infinite chain through weak Ag\u22efO inter\u00adactions [2.765\u2005(2)\u2005\u00c5], Ag\u22efAg inter\u00adactions [3.1511\u2005(4)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.623\u2005(4)\u2005\u00c5]. N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds assemble these chains into a three-dimensional network.In the title complex, [Ag(NO DOI: 10.1107/S1600536809019138/hy2199Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Both Ag+ cations show a distorted square-pyramidal coordination. Ag1 is bonded to two water molecules, one L N atom, one N atom and one carboxylate O atom from a neighbouring L, whereas Ag2 is surrounded by two L N atoms, two L carboxylate O atoms and one bridging water molecule. O\u2014H\u22efO hydrogen-bonding inter\u00adactions involving water clusters and carboxyl\u00adate O atoms link the mol\u00adecules into a three-dimensional supra\u00admolecular architecture, which is further consolidated by weak C\u2014H\u22efO inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance 3.643\u2005(5)\u2005\u00c5].In the title silver(I) coordination polymer, {[Ag DOI: 10.1107/S160053680802984X/kj2095Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the two ligands, the pyridyl rings form dihedral angles of 21.0\u2005(1) and 15.5\u2005(1)\u00b0. The crystal packing exhibits an extensive network of O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions proved by short distances of 3.650\u2005(1) and 3.732\u2005(2)\u2005\u00c5 between the centroids of pyridyl rings of neighbouring mol\u00adecules.In the title compound, [Cu(C DOI: 10.1107/S1600536809009453/cv2529Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal packing is stabilized by inter\u00admolecular O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds. A weak C\u2014H\u22ef\u03c0 inter\u00adaction also occurs.In the title compound, C DOI: 10.1107/S1600536809055391/jj2016Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The OH group of the 2-hydroxy\u00adbenzoate anion is disordered over two positions with site-occupancy factors of 0.5. The asymmetric unit is completed with by an uncoordinated half-mol\u00adecule of dmphen, disordered about a crystallographic twofold axis. In the crystal structure, mol\u00adecules are linked into a two-dimensional framework by O\u2014H\u22efN, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. The packing of the structure is further stabilized by \u03c0\u2013\u03c0 stacking inter\u00adactions involving dmphen mol\u00adecules, with centroid\u2013centroid separations of 3.8027\u2005(3) and 3.6319\u2005(3)\u2005\u00c5.In the asymmetric unit of the title complex, [Mn(C DOI: 10.1107/S1600536809000981/bh2207Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There is a weak \u03c0\u2013\u03c0 stacking inter\u00adaction between symmetry-related triazolyl rings with a centroid\u2013centroid distance of 3.802\u2005(4)\u2005\u00c5 and a perpendicular distance of 3.413\u2005\u00c5 between the rings.In the title complex, [Zn(C DOI: 10.1107/S160053680903877X/bt5062Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each NiII ion has a distorted square-pyramidal environment, with the basal plane formed by three O [Ni\u2014O = 1.9290\u2005(16)\u20131.9588\u2005(10)\u2005\u00c5] and one N [Ni\u2014N = 1.9828\u2005(18)\u2005\u00c5] atoms and the apical position occupied by the water mol\u00adecule [Ni\u2014O = 2.2643\u2005(11)\u2005\u00c5]. The water mol\u00adecules are involved in the formation of O\u2014H\u22efO hydrogen bonds.In the title compound, {[Ni DOI: 10.1107/S1600536808031619/cv2458Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II compound, [Zn2(C12H9O3)4(H2O)2], is centrosymmetric. Each Zn atom is coordinated by two bridging 2-naphthoxyacetate anions, one terminal 2-naphth\u00adoxy\u00adacetate anion and one water mol\u00adecule in a distorted ZnO4 tetra\u00adhedral geometry. The naphthalene system of the bridging ligand is nearly perpendicular to the naphthalene of the terminal ligand, with a dihedral angle of 78.26\u2005(6)\u00b0. Within the binuclear mol\u00adecule the Zn\u22efZn separation is 3.815\u2005(5)\u2005\u00c5. In the crystal structure, inter\u00admolecular O\u2014H\u22efO hydrogen bonding between the water mol\u00adecule and carboxyl\u00adate groups helps to stabilize the crystal structure.The title binuclear Zn DOI: 10.1107/S1600536809013750/xu2494Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "III compound, [In2(SO4)3(C12H8N2)2(H2O)2]\u00b72H2O, each InIII cation is coordinated by a 1,10-phenanthroline (phen) ligand, a water mol\u00adecule and three sulfate O atoms in a distorted InN2O4 octa\u00adhedral geometry. Three sulfate anions bridge two InIII cations, forming the dinuclear entities. O\u2014H\u22efO and weak C\u2014H\u22efO hydrogen bonding is observed in the crystal structure. The crystal structure is further consolidated by \u03c0\u2013\u03c0 stacking between nearly parallel phen ring systems [dihedral angle = 4.2\u2005(4)\u00b0], the centroid\u2013centroid distance between benzene rings of adjacent phen ligands being 3.528\u2005(9)\u2005\u00c5.In the title dinuclear In DOI: 10.1107/S1600536810036330/xu5026Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit contains one copper cation to which two acetate units bind asymmetrically; one of the Cu\u2014O distances is rather long [2.740\u2005(2)\u2005\u00c5], almost at the limit of coordination. These Cu\u2014O bonds define an equatorial plane to which the Cu\u2014N bonds to the INA ligands are almost perpendicular, the Cu\u2014N vectors subtending angles of 2.4\u2005(1) and 2.3\u2005(1)\u00b0 to the normal to the plane. The metal coordination geometry can be described as a slightly distorted trigonal bipyramid if the extremely weak Cu\u2014O bond is disregarded, or as a highly distorted square bipyramid if it is not. The double acetate bridge between the copper ions is not coplanar with the CuO4 equatorial planes, the dihedral angle between the (O\u2014C\u2014O)2 and O\u2014Cu\u2014O groups being 34.3\u2005(1)\u00b0, resulting in a sofa-like conformation for the 8-member bridging loop. In the crystal, N\u2014H\u22efO hydrogen bonds occur, some of which generate a head-to tail-linkage between INA units, giving raise to chains along [101]; the remaining ones make inter-chain contacts, defining a three-dimensional network. There are in addition a number of C\u2014H\u22efO bonds involving aromatic H atoms. Probably due to steric hindrance, the aromatic rings are not involved in significant \u03c0\u22ef\u03c0 inter\u00adactions.The title centrosymmetric bimetallic complex, [Cu For coo al. 2003. For a c al. 2007. 2(C2H3O2)4(C6H6N2O)4] = 0.039 wR(F 2) = 0.099 S = 1.08 3736 reflections244 parametersH-atom parameters constrainedmax = 0.36 e \u00c5\u22123 \u0394\u03c1min = \u22120.43 e \u00c5\u22123 \u0394\u03c1 MSC/AFC Diffractometer Control Software used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008SHELXTL-NT (Sheldrick, 2008SHELXTL-NT and PLATON (Spek, 2009Data collection: 10.1107/S1600536810004393/hb5326sup1.cif Crystal structure: contains datablocks I, New_Global_Publ_Block. DOI: 10.1107/S1600536810004393/hb5326Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal packing is stabilized by inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances 3.572\u2005(1) and 3.671\u2005(1)\u2005\u00c5 together with C\u2014H\u22efBr hydrogen bonds.The title compound, [CdBr DOI: 10.1107/S1600536809020352/sj2628Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Longer Cu\u22efO contacts [2.680\u2005(2)\u2005\u00c5] complete an approximate square-based pyramidal coordination geometry around CuII, forming a dimeric complex across a centre of inversion. The dimeric complexes form stacks along the a axis, with Cu\u22efO contacts of 3.332\u2005(2)\u2005\u00c5 between them. The Na+ cations and perchlorate anions lie on twofold rotation axes between the stacks. The former are coordinated by two disordered water mol\u00adecules , and form Na\u22efO contacts of 3.698\u2005(3)\u2005\u00c5 to the perchlorate anions and Na\u22ef\u03c0 contacts to neighbouring salicylideneglycinate ligands [shortest Na\u22efC = 3.516\u2005(3)\u2005\u00c5].In the title compound, [Na(H DOI: 10.1107/S1600536808039561/bi2318Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "N-heterocycle in the title centrosymmetric dinuclear compound, [Ag2(NO3)2(C8H15N3)2], bridges two metal atoms through its imino N atoms. The N\u2014Ag\u2014N skeleton is bent [N\u2014Ag\u2014N = 127.2\u2005(3)\u00b0]; as one of two O atoms of the nitrate anion is nearly coplanar with this N\u2014Ag\u2014N skeleton [Ag\u2014O = 2.63\u2005(1)\u2005\u00c5], the coordination geometry around the AgI atom is regarded as trigonal-planar. One of the two isopropyl groups is disordered over two positions in respect of the methyl groups in a 1:1 ratio. In the crystal structure, inter\u00admolecular N\u2014H\u22efO hydrogen bonding is observed between the nitrate groups and triazole ligands.The neutral DOI: 10.1107/S1600536809028384/xu2560Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by inter\u00admolecular C\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions with a centroid\u2013centroid distance of 3.6136\u2005(16)\u2005\u00c5.In the title mononuclear zinc(II) complex, [Zn(C DOI: 10.1107/S1600536809033893/rk2159Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds link the cations, anions and water mol\u00adecules into a three-dimensional supra\u00admolecular structure. The crystal packing also exhibits inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings of the anions with a centroid\u2013centroid distance of 3.604\u2005(2)\u2005\u00c5.The asymmetric unit of the title compound, [Mg(H DOI: 10.1107/S160053680904344X/cv2632Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The 2-amino\u00adethyl group has a coiled conformation with an N\u2014C\u2014C\u2014NH2 torsion angle of 53.6\u2005(4)\u00b0. In the crystal structure, inter\u00admolecular N\u2014H\u22efN and weak C\u2014H\u22efO hydrogen bonds link mol\u00adecules into chains along [001].In the title compound, C DOI: 10.1107/S160053680901962X/lh2821Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Zn\u2014Br bond lengths range from 2.400\u2005(2) to 2.408\u2005(3)\u2005\u00c5 and the Br\u2014Zn\u2014Br angles range from 108.14\u2005(6) to 115.15\u2005(15)\u00b0. In the crystal structure, the [ZnBr4]2\u2212 anion is connected to two cations through N\u2014H\u22efBr and H2C\u2014H\u22efBr hydrogen bonds, forming two-dimensional cation\u2013anion\u2013cation layers normal to the b axis. No significant Br\u22efBr inter\u00adactions [the shortest being 4.423\u2005(4)\u2005\u00c5] are observed in the structure.In the crystal structure of the title compound, (C DOI: 10.1107/S1600536809015219/at2771Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Edge-to-face C\u2014H\u22ef\u03c0 inter\u00adactions exist between mol\u00adecules, with a dihedral angle of 37.8\u2005(1)\u00b0 between the benzene ring planes and a shortest H\u22efcentroid distance of 3.62\u2005(5)\u2005\u00c5.In the title compound, [ZnCl DOI: 10.1107/S1600536808037860/bi2311Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII atoms in each complex adopt a distorted square-pyramidal coordination geometry being penta\u00adcoordinated by one N and two O atoms of 4-hydroxy\u00adpyridine-2,6-dicarboxyl\u00adate anions and two N atoms of 2,9-dimethyl-1,10-phenanthroline (dmp) molecules. In the crystal structure, there are O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and five \u03c0\u2013\u03c0 stacking inter\u00adactions with centroid\u2013centroid distances in the range 3.620\u2005(1)\u20133.712\u2005(1)\u2005\u00c5. In addition, a C\u2014H\u22ef\u03c0 inter\u00adaction between a heterocyclic ring of dmp is observed to reinforce the crystal cohesion.In the title complex, [Cu(C DOI: 10.1107/S1600536809021588/pv2164Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "These components are inter\u00adconnected in the crystal structure by an extensive network of O\u2014H\u22efO, N\u2014H\u22efO, C\u2014H\u22efO, O\u2014H\u22efN, O\u2014H\u22efCl, N\u2014H\u22efCl and C\u2014H\u22efCl hydrogen bonds. The shortest inter\u00admolecular inter\u00adaction is realized between the cation and anion . C\u2014H\u22efO inter\u00adactions also play a important role in the inter\u00adconnection of the cations.The asymmetric unit of the title compound, (C DOI: 10.1107/S160053681000615X/rk2191Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the complex mol\u00adecule, the two benzimidazole planes are twisted, making a dihedral angle of 55.93\u2005(11)\u00b0. The three-dimensional framework is organized by inter\u00admolecular N\u2014H\u22efO hydrogen bonding and O\u2014H\u22efO inter\u00adactions and \u03c0\u2013\u03c0 inter\u00adactions between adjacent benzimidazole rings [centroid\u2013centroid distance = 3.586\u2005(3)\u2005\u00c5].In the title compound, [Zn(C DOI: 10.1107/S1600536810015631/kp2256Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular O\u2014H\u22efN inter\u00adactions link the monomeric units into a two-dimensional hydrogen-bonded network, which is extended to a three-dimensional supra\u00admolecular aggregate via \u03c0\u22ef\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances 3.809\u2005(3) and 3.499\u2005(3)\u2005 \u00c5].In the mol\u00adecular structure of the centrosymmetric mononuclear complex [Ni(2-bpt) DOI: 10.1107/S1600536809012598/hg2489Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of neighbouring mol\u00adecules [centroid\u2013centroid distance = 3.698\u2005(4)\u2005\u00c5], and by C\u2014H\u22ef\u03c0 inter\u00adactions between a methyl\u00adene H atom of the butyl group and the benzene ring of the benzofuran system. Additionally, the crystal structure exhibits weak inter\u00admolecular C\u2014H\u22efO contacts. The butyl group is disordered over two positions, with site-occupancy factors, from refinement, of 0.720\u2005(8) and 0.280\u2005(8).In the title compound, C DOI: 10.1107/S1600536808043985/tk2347Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "This compound is composed of an anionic complex, [Cr(pydc)2]\u2212, protonated 4,4\u2032-bipyridine as a counter-ion, +, and four uncoordinated water mol\u00adecules. The anion is a six-coordinate complex with a distorted octa\u00adhedral geometry around the CrIII atom, formed by two tridentate pyridine-2,6-dicarboxyl\u00adate, pydc2\u2212, groups. Inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, and C\u2014O\u22ef\u03c0 stacking inter\u00adactions (with distances of 3.3390\u2005(13) and 3.4575\u2005(13)\u2005\u00c5) connect the various components into a supra\u00admolecular structure.The title compound, (C DOI: 10.1107/S1600536808006594/om2216Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Fe2S2 unit exhibits a butterfly conformation and the ferrocenyldiphenyl\u00adphosphine ligand is trans to the Fe\u2014Fe bond. The Fe\u2014Fe distance of 2.5160\u2005(8)\u2005\u00c5 is longer than found in related model structures. Intra\u00admolecular C\u2014H\u22efS and inter\u00admolecular C\u2014H\u22efO hydrogen bonds are observed.The title compound, [Fe DOI: 10.1107/S1600536808032698/ci2685Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The water mol\u00adecules and the 1-(4-chloro\u00adphen\u00adyl)piperazine-1,4-diium cations inter\u00adact with the [ZnCl4]2\u2212 anions through O\u2014H\u22efCl, N\u2014H\u22efCl, N\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds (five simple and one bifurcated). Inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions are present between adjacent aromatic rings of 1-(4-chloro\u00adphenyl)\u00adpiperazine-1,4-diium cations (the centroid\u2013centroid distance is 3.453\u2005\u00c5).In the crystal structure of the title compound, (C DOI: 10.1107/S1600536808016590/bg2190Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each Au atom is in a square-planar environment coordinated by diphenyl\u00adphosphanide and 2,6-diphenyl\u00adpyridine ligands. There are weak \u03c0\u2013\u03c0 stacking inter\u00adactions between neighbouring mol\u00adecules (the inter\u00adplanar separations between two neighbouring dpp units are 3.40 and 3.57\u2005\u00c5). The intra\u00admolecular Au\u22efAu separation is 3.788\u2005(5)\u2005\u00c5. The crystal structure shows weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds involving an O atom of the perchlorate counter-ion and the N atom of the acetonitrile solvent mol\u00adecule, respectively.The title complex, [Au DOI: 10.1107/S1600536808024537/bx2152Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The structure contains infinite chains of ZnCl2 units connected by the bifunctional thia\u00addiazole ligands, with ZnII adopting a distorted tetra\u00adhedral coordination geometry. The dihedral angle between the two pyridyl rings in each ligand is 34.3\u2005(1)\u00b0, and the dihedral angles between the thia\u00addiazole ring and the two pyridyl rings are 18.3\u2005(2) and 16.1\u2005(2)\u00b0. The shortest Zn\u22efZn distance within each polymeric chain is 11.862\u2005(3)\u2005\u00c5, while the shortest inter\u00adchain Zn\u22efZn distance is 7.057\u2005(3)\u2005\u00c5.The title compound, [ZnCl DOI: 10.1107/S1600536808001967/bi2274Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the anion, the S\u2014O bond [1.551\u2005(2)\u2005\u00c5] involving the O atom bearing the acid H atom is longer than the other three S\u2014O bonds, which range from 1.437\u2005(1) to 1.454\u2005(1)\u2005\u00c5.The crystal structure of the title salt, C DOI: 10.1107/S1600536809048533/xu2677Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Intra\u00admolecular C\u2014H\u22efO hydrogen bonds result in the formation of two planar and two non-planar five-membered rings; the latter adopt envelope conformations. There are weak \u03c0\u2013\u03c0 inter\u00adactions between aromatic rings, with centroid-to-centroid distances of 3.796\u2005(2) and 4.171\u2005(2)\u2005\u00c5. There is also a single C\u2014Cl\u22ef\u03c0 inter\u00adaction .In the mol\u00adecule of the title compound, [Sn(C DOI: 10.1107/S1600536808018321/hk2473Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Ni\u2014O(phenolate) bond [1.9750\u2005(16)\u2005\u00c5] is shorter than the Ni\u2014O(carbon\u00adyl) bond [2.0840\u2005(16)\u2005\u00c5] and the Ni\u2014N bonds (mean 2.120\u2005\u00c5).In the title compound, [Ni(C DOI: 10.1107/S1600536808023489/is2292Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "I atom in the salt, [Ag(C4H6N2)2]NO3\u00b72H2O, shows a nearly linear coordination [N\u2014Ag\u2014N = 178.26\u2005(7)\u00b0]. The cation, anion and water mol\u00adecules are linked by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds into a layer motif extending parallel to (101).The Ag DOI: 10.1107/S1600536809045838/bt5122Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The HfO8 coordination polyhedron shows a slightly distorted Archimedean square anti-prismatic coordination with average Hf\u2014O, C\u2014O, C\u2014CMe distances of 2.19\u2005(2), 1.26\u2005(2) and 1.49\u2005(2)\u2005\u00c5, respectively, and an O\u2014Hf\u2014O bite angle of 75.3\u2005(5)\u00b0. Weak O\u2014H\u22efO hydrogen bonding inter\u00adactions are observed between one of the bridging hydr\u00adoxy groups and the disordered solvent mol\u00adecule.The binuclear title compound, [Hf DOI: 10.1107/S1600536809041658/wm2265Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Cl ligands are trans to each other, with a Cl\u2014Pt\u2014Cl angle of 178.83\u2005(8)\u00b0. The pyridine ring forms a dihedral angle of 48.8\u2005(2)\u00b0 with the planar PtCl2SN unit. Within the mol\u00adecule, the distance between Pt atoms is 20.262\u2005(5)\u2005\u00c5 and the N\u22efN separation between the terminal pyridyl rings is 16.23\u2005(1)\u00c5.The title dinuclear platinum compound, [Pt DOI: 10.1107/S1600536808024914/gk2157Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Mol\u00adecules are connected into layers in the ac plane via extensive C\u2014H\u22efCl contacts as each Cl atom forms two such inter\u00adactions. Contacts between the layers are of the type C=O\u22ef\u03c0 [O\u22efcentroid distance = 3.110\u2005(8)\u2005\u00c5].In the title compound, [HgCl DOI: 10.1107/S1600536809049289/hy2254Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The CuII atom is coordinated by one tridentate N-salicylideneglycinate Schiff base ligand, one 4-methyl\u00adquinoline ligand and one water mol\u00adecule, leading to a slightly distorted square-pyramidal N2O3 geometry. In the crystal structure, the mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds into linear chains in the [100] direction. The structure is further stabilized by inter\u00admolecular C\u2014H\u22efO inter\u00adactions and C\u22efC contacts with C\u22efC = 3.3058\u2005(2), 3.3636\u2005(2) and 3.3946\u2005(2)\u2005\u00c5.The title complex, [Cu(C DOI: 10.1107/S1600536807067852/tk2236Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Co(C8H5O3)2(C10H14N2O)2(H2O)2], the metal centre is located on an inversion center and is coordinated by two 4-formyl\u00adbenzoate (FOB), two diethyl\u00adnicotinamide (DENA) ligands and two water mol\u00adecules in a slightly distorted CoO4N2 octa\u00adhedral geometry. In the crystal structure, O\u2014H\u22efO hydrogen bonds link the mol\u00adecules into infinite chains. \u03c0\u2013\u03c0 contacts between the parallel pyridine rings of neighboring DENA ligands [centroid\u2013centroid distance = 3.652\u2005(3)\u2005\u00c5] further stabilize the crystal structure.In the crystal structure of the title Co DOI: 10.1107/S1600536809008265/xu2489Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "A Cu\u22efCu separation of 3.2281\u2005(3)\u2005\u00c5 is observed. The in-plane Cu\u2014Ophenolate distance [1.9342\u2005(8)\u2005\u00c5] is shorter than the axial distance [2.252\u2005(8)\u2005\u00c5]. The Cu\u2014Namine and Cu\u2014Npy distances are similar . The Cu\u2014Nthio\u00adcyanate distance [1.9678\u2005(11)\u2005\u00c5] is in the range found for Cu\u2014N distances in previously determined structures containing coordinated thio\u00adcyanate anions. There is an inter\u00admolecular hydrogen bond between the amine H atom and the S atom of a coordinated thio\u00adcyanate anion.The centrosymmetric binuclear complex, [Cu DOI: 10.1107/S1600536809031742/kp2227Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the structure, the ZrIV atom is nine-coordinated by three (pydc)2\u2212 groups, resulting in an anionic complex which is balanced by two (tataH)+ cations. One of the NH2 groups shows positional disorder, with site occupation factors of 0.60 and 0.40. There are four uncoordinated water mol\u00adecules (one of which is disordered with occupation factors of 0.70 and 0.30) in the crystal structure. Several inter\u00admolecular inter\u00adactions, including O\u2014H\u22efO, O\u2014H\u22efN, N\u2014H\u22efO, N\u2014H\u22efN, C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds, a C\u2014O\u22ef\u03c0 inter\u00adaction , and \u03c0\u2013\u03c0 stacking [with centroid\u2013centroid distances of 3.694\u2005(2) and 3.802\u2005(2)\u2005\u00c5] are also present.The title compound, (C DOI: 10.1107/S1600536808029887/om2259Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The hydroxonium hydrate (H5O2)+, also known as the Zundel cation, resides on a twofold rotation axis. The O\u2014H distance is 1.274\u2005(14)\u2005\u00c5, the O\u22efO distance is 2.518\u2005(5)\u2005\u00c5, and the O\u2014H\u2014O angle is 162\u2005(8)\u00b0. One of the melamine H+ cations, the uncoordinated pydcH2, and two water mol\u00adecules also reside on crystallographic twofold axes. The CuII atom has a tetra\u00adgonally distorted octa\u00adhedral coordination environment. The structure features extensive hydrogen bonding, with 21 distinct inter\u00adactions. There is also a centrosymmetric C=O\u22ef\u03c0 inter\u00adaction with an O\u22efcentroid distance of 3.288\u2005(3)\u2005\u00c5. The structure is similar to a mixed-valence manganese(II/III) structure but shows inter\u00adesting differences in the metal-atom coordination. One of the water molecules is equally disordered with respect to a twofold axis.The reaction of copper(II) nitrate hexa\u00adhydrate with pyridine-2,6-dicarboxylic acid (pydcH DOI: 10.1107/S1600536809000828/gw2056Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions. In addition, the crystal structure exhibits a Br\u22ef\u03c0 inter\u00adaction of 3.551\u2005(3)\u2005\u00c5 between the Br atom and the centroid of the benzene ring of an adjacent mol\u00adecule.In the title compound, C DOI: 10.1107/S1600536809008101/zl2184Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The 2,5-dimethyl\u00adpyrazine 1,4-dioxide ligand, which also lies on an inversion center, acts as a bridging ligand, linking symmetry-related CoII ions [Co\u22efCo = 8.669\u2005(3)\u2005\u00c5] and forming one-dimensional chains along the b axis. In the crystal structure, these chains are linked by inter\u00admolecular aqua\u2013perchlorate O\u2014H\u22efO hydrogen bonds, forming two-dimensional layers which are in turn connected into a three-dimensional network via \u03c0\u2013\u03c0 stacking inter\u00adactions between quinoline rings, with a centroid\u2013centroid distance of 3.580\u2005(3)\u2005\u00c5. An intermolecular O\u2014H\u22efCl inter\u00adaction is also present.In the title complex, {[Co(C DOI: 10.1107/S1600536810008895/lh5007Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The AgI ion is linearly coordinated by two dipyrazin-2-ylamine ligands through two pyrazine N atoms. (ClO4 \u2212)\u22ef\u03c0(pyrazine) [O\u22efcentroid distances of 3.612\u2005(3) and 3.664\u2005(1)\u2005\u00c5] and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.518\u2005(2)\u2005\u00c5] as well as O\u2014H\u22efO and N\u2014H\u22efO hydrogen-bonds assemble the chains into a three-dimensional supra\u00admolecular aggregation. In the title complex, {[Ag(C DOI: 10.1107/S1600536809037532/bg2275Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, inversion-related mol\u00adecules are linked into dimers by O\u2014H\u22efO hydrogen bonds. The packing is further stabilized by \u03c0\u2013\u03c0 inter\u00adactions involving the benzene rings of the dmphen and hydroxy\u00adbenzoate units, with centroid\u2013centroid distances of 3.4930\u2005(14) or 3.5727\u2005(14)\u2005\u00c5.In the title compound, [Cu(C DOI: 10.1107/S1600536808034788/ci2692Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The HgII atoms show a typical linear geometry to a C atom of the benzene ring and to a Cl atom. A benzene C and the azomethine N atom chelate the HgII atoms with weak intra\u00admolecular Hg\u22efN bonding distances of 2.735\u2005(3) and 2.739\u2005(3)\u2005\u00c5, respectively. The resulting five-membered metallacycles are nearly coplanar with the benzene rings [dihedral angles = 0.9\u2005(1) and 0.7\u2005(1)\u00b0], while the pyridine rings make dihedral angles with the benzene units of 58.17\u2005(1) and 56.58\u2005(1)\u00b0. In the crystal structure, the HgII complexes are linked by hydr\u00adoxy donor and pyridine acceptor groups into chains along [010]. The water mol\u00adecules connect the complexes through inter\u00admolecular O\u2014H\u22efOcarbon\u00adyl bonds in the a-axis direction, and the azomethine H atoms donate towards the water O atoms, forming a three-dimensional network of inter\u00admolecular O\u2014H\u22efN, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds.The asymmetric unit of the title compound, [Hg(C Cl]\u00b7H2O = 0.021 wR(F 2) = 0.046 S = 1.05 5658 reflections381 parametersH-atom parameters constrainedmax = 0.52 e \u00c5\u22123 \u0394\u03c1min = \u22120.83 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008SHELXTL (Sheldrick, 2008SHELXTL and PLATON (Spek, 2009Data collection: 10.1107/S1600536809023824/si2184sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536809023824/si2184Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Mn3+ ions are bridged by anionic pyridine-2-carboxyl\u00adate (pic) ligands, thereby forming a chain-like structure along the c axis, and are six-coordinated in a distorted octa\u00adhedral environment by two O atoms of the two different carboxyl\u00adate groups, two O atoms of two water mol\u00adecules and two Br atoms. The complex displays inter\u00admolecular O\u2014H\u22efBr, O\u2014H\u22efN, O\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efBr hydrogen bonding. There may also be inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between adjacent pyridine rings, with a centroid\u2013centroid distance of 3.993\u2005(8)\u2005\u00c5.The asymmetric unit of the title compound, [MnBr DOI: 10.1107/S160053680902844X/im2127Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecular conformation is stabilized by weak intra\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds. The crystal structure features weak C\u2014H\u22ef\u03c0 inter\u00adactions.In the title mol\u00adecule, C DOI: 10.1107/S1600536809037830/is2462Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, a combination of close contacts formed via Ag\u22efN inter\u00adactions [Ag\u22efN = 3.098\u2005(2) and 3.261\u2005(2)\u2005\u00c5] from symmetry-related mol\u00adecules and inter\u00admolecular N\u2014H\u22efO hydrogen bonds between CF3CO2 \u2212 anions and the hydrazone groups of two ligands give rise to chains. Furthermore, there are Ag\u22efO inter\u00adactions with a separation of 2.765\u2005(2)\u2005\u00c5 between chains. The F atoms of the CF3CO2 \u2212 anion are disordered over two sites with refined occupancies of 0.593\u2005(5) and 0.407\u2005(5).In the title compound, [Ag(C DOI: 10.1107/S1600536809029183/lh2867Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ansa-molybdocene [C2Me4(\u03b75-C5H4)2]Mo unit has a typical bent-sandwich metallocene structure with an inter-ring angle of 127.98\u2005(8)\u00b0. The Mo atom in the bridging (\u03bc2-O)(\u03bc3-O)2MoO2 group has a distorted trigonal\u2013bipyramidal coordination. The Mo\u2014(\u03bc3-O) and Mo\u2014(\u03bc2-O) bond distances inside the units are slightly longer than the Mo\u2014(\u03bc3-O) bond distance between the units [1.9986\u2005(14)\u2005\u00c5]. The solvent water mol\u00adecules together with complex O atoms form a network of O\u2014H\u22efO hydrogen bonds.The title compound, [Mo DOI: 10.1107/S1600536808031668/hb2806Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "It is composed of a planar r.m.s. deviation (0.0012\u2005\u00c5) tetra\u00adzole ring which is nearly coplanar with the benzene ring, the dihedral angle being 2.67\u2005(9)\u00b0. In the crystal, symmetry-related mol\u00adecules are linked by inter\u00admolecular N\u2014H\u22efN hydrogen bonds. The mol\u00adecules stack along [100] with a \u03c0\u22ef\u03c0 inter\u00adaction involving the phenyl and tetra\u00adzole rings of adjacent mol\u00adecules [centroid\u2013centroid distance = 3.5639\u2005(15)\u2005\u00c5]. The H atom of the N\u2014H group is disordered over two sites of equal occupancy. The methyl H atoms were modelled as disordered over two sets of sites of equal occupancy rotated by 60\u00b0 with respect to each other.The title compound, C DOI: 10.1107/S1600536809036411/su2134Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Ni(C7H4ClO2)2(C6H6N2O)2(H2O)2], is centrosymmetric with the Ni atom located on an inversion centre. The mol\u00adecule contains two 2-chloro\u00adbenzoate (CB) and two nicotinamide (NA) ligands and two water mol\u00adecules, all ligands being monodentate. The four O atoms in the equatorial plane around the Ni atom form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl group and the adjacent benzene ring is 29.48\u2005(16)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 83.16\u2005(5)\u00b0. In the crystal structure, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into infinite chains. \u03c0\u2013\u03c0 Contacts between the benzene and pyridine rings [centroid\u2013centroid distance = 3.952\u2005(1)\u2005\u00c5] may further stabilize the crystal structure. There is also a C\u2014H\u22ef\u03c0 inter\u00adaction.The title Ni DOI: 10.1107/S1600536809011209/xu2501Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure stabilized by three different inter\u00admolecular non-classical C\u2014H\u22efO hydrogen bonds. The crystal structure also exhibits aromatic \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of adjacent benzofuran ring systems [centroid\u2013centroid distance = 3.688\u2005(2)\u2005\u00c5]In the title compound, C DOI: 10.1107/S1600536809037003/bq2158Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angles between the benzimidazole ring system and the phenyl rings in each of the benzyl\u00adbenzimidazole units are 78.56\u2005(12), 81.68\u2005(11), 75.76\u2005(10) and 85.78\u2005(9)\u00b0. In the crystal structure, there are weak but significant inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions with centroid\u2013centroid distances of 3.685\u2005(1) and 3.978\u2005(1)\u2005\u00c5.In the title compound, [Zn(C DOI: 10.1107/S1600536809022296/lh2840Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The thia\u00addiazole rings are oriented at a dihedral angle of 84.87\u2005(4)\u00b0. Intra\u00admolecular N\u2014H\u22efCl inter\u00adactions result in the formation of two six-membered rings having envelope and planar conformations. In the crystal structure, inter\u00admolecular N\u2014H\u22efN and N\u2014H\u22efCl inter\u00adactions link the mol\u00adecules into a three-dimensional network. \u03c0\u2013\u03c0 contacts between thia\u00addiazole rings [centroid\u2013centroid distance = 3.602\u2005(1)\u2005\u00c5] may further stabilize the structure.In the title compound, [ZnCl DOI: 10.1107/S1600536809032073/hk2748Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "It is an example of a new type of tropolone derivative that has not been characterized via solid-state methods. Weak intra\u00admolecular C\u2014H\u22efN and inter\u00admolecular C\u2014H\u22efO hydrogen bonds, and \u03c0\u2013\u03c0 stacking inter\u00adactions between the tropolone rings [centroid\u2013centroid distance = 3.590\u2005(8)\u2005\u00c5] are observed in the crystal structure.In the title compound, [Rh(C DOI: 10.1107/S160053680803780X/hy2163Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the ferrocene unit, the substituted (Cps) and unsubstituted (Cp) cyclo\u00adpenta\u00addienyl rings are eclipsed and almost parallel [Cps\u2014Fe\u2014Cps angle = 176.1\u2005(2)\u00b0]. The mol\u00adecule is linked into an S(5) motif via intra\u00admolecular C\u2014H\u22efO hydrogen bonds. The mol\u00adecules are arranged into a three-dimensional framework by five inter\u00admolecular C\u2014H\u22efO hydrogen bonds and one inter\u00admolecular C\u2014H\u22ef\u03c0(Cps) inter\u00adaction.In the title compound, [Fe(C DOI: 10.1107/S1600536808025518/pv2096Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each PZDC2\u2212 dianion serves as a spacer, connecting adjacent metal atoms into a polymeric chain structure parallel to the b axis. The chain motif is consolidated into a three-dimensional supra\u00admolecular network by O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 aromatic stacking inter\u00adactions involving adjacent DPPZ ligands and PZDC2\u2212 dianions with centroid\u2013centroid separations of 3.522\u2005(6) and 3.732\u2005(8)\u2005\u00c5, respectively.In the title compound, [Zn(C DOI: 10.1107/S1600536808022824/rz2235Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the complex, the two MnIII ions are bridged by two O atoms from two symmetry-related N,N\u2032-bis\u00ad-1,2-diimino\u00adbenzene dianionic ligands with the longer Mn\u2014O distance of 2.703\u2005(3)\u2005\u00c5, thus each Mn ion is six-coordinated by two N and three O atoms from the two dianionic ligands and one capping Cl atom in a distorted octa\u00adhedral environment. The crystal structure displays inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between adjacent benzene rings, with a shortest centroid\u2013centroid distance of 3.673\u2005(2)\u2005\u00c5, and inter\u00admolecular C\u2014H\u22efO, C\u2014H\u22ef Cl and C\u2014H\u22ef Br hydrogen bonds.The asymmetric unit of the title compound, [Mn DOI: 10.1107/S1600536810003247/xu2722Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each CuII atom is coordinated by two O atoms and one N atom from the tridentate ligand L 2\u2212 [LH2 = (E)-2--2-(4-hydroxy\u00adphenyl)acetic acid] and the O atom of a dimethyl\u00adformamide mol\u00adecule to give a slightly distorted square-planar geometry. The two unique mol\u00adecules form a dimer through weak C\u2014H\u22efO hydrogen bonds. In the dimer, the Cu\u22efCu distance is 3.712\u2005(1)\u2005\u00c5. In the crystal structure, mol\u00adecules form a one-dimensional chain through C\u2014H\u22efO hydrogen bonds. These are further aggregated into a three-dimensional network by O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds.In the title complex, [Cu(C DOI: 10.1107/S1600536808007915/sj2468Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There are two intra\u00admolecular N\u2014H\u22efBr hydrogen bonds completing twisted six-membered rings with R(6) motifs. The dihedral angles between the aromatic rings in the ligands are 62.11\u2005(13) and 85.73\u2005(13)\u00b0. In the crystal, components are linked by N\u2014H\u22efO, O\u2014H\u22efS and O\u2014H\u22ef\u03c0 inter\u00adactions. There also exist \u03c0\u2013\u03c0 inter\u00adactions with a distance of 3.876\u2005(2)\u2005\u00c5 between the centroids of benzene rings of two different ligands. Together, the inter\u00admolecular inter\u00adactions lead to a three-dimensional network.In the title compound, [CuBr(C DOI: 10.1107/S1600536809026038/hb5021Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "All hydroxyl groups act as donors and acceptors in hydrogen bonding and the mol\u00adecule is involved in ten inter\u00admolecular O\u2014H\u22efO inter\u00adactions [O\u22efO = 2.672\u2005(5)\u20132.776\u2005(4)\u2005\u00c5] with eight neighbouring mol\u00adecules. Two independent O\u2014H\u22efO\u2014H\u22ef helices extending along the z axis are found in this structure.The mol\u00adecule of the title compound, C DOI: 10.1107/S1600536809000397/gk2181Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "It contains a half-sandwich complex of ruthenium with a three-legged piano-stool geometry, with Ru\u2014P = 2.3585\u2005(4) and 2.3312\u2005(4)\u2005\u00c5, and Ru\u2014N = 2.0422\u2005(15)\u2005\u00c5\u2005as the legs. The CF3SO3 \u2212 anion is anchored in the crystal lattice by C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds, with C\u22efF,O distances starting at 3.125\u2005(2)\u2005\u00c5.The title compound, [Ru(C DOI: 10.1107/S1600536809038720/ez2188Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The slightly distorted octa\u00adhedral environment of the Ru center is formed by four P atoms [Ru\u2014P = 2.4417\u2005(6) and 2.4544\u2005(6)\u2005\u00c5] from two different -TangPhos ligands and two Cl atoms [Ru\u2014Cl = 2.4267\u2005(5)\u2005\u00c5].In the title compound, [RuCl DOI: 10.1107/S1600536808008301/cv2390Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "At the core of the structure is an Li2O2 four-membered ring. The structure is centrosymmetric with an inversion centre midway between two Li atoms. Inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 inter\u00adactions between the 2-methyl\u00adpyridine rings exist [centroid\u2013centroid distance = 3.6312\u2005(16)\u2005\u00c5].The title compound, [Li DOI: 10.1107/S1600536808042748/gw2055Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Ni(C7H4BrO2)2(C6H6N2O)2(H2O)2], is centrosymmetric. It contains two 2-bromo\u00adbenzoate (BB) ligands, two nicotinamide (NA) ligands and two water mol\u00adecules, all of them being monodentate. The four O atoms in the equatorial plane around the Ni atom form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 30.81\u2005(17)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 84.66\u2005(6)\u00b0. In the crystal structure, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a supra\u00admolecular structure. A weak C\u2014H\u22ef\u03c0 inter\u00adaction is also found.The title Ni DOI: 10.1107/S1600536809021710/xu2538Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII ion is coordinated by two N atoms [Cu\u2014N = 1.960\u2005(4)\u2005\u00c5] and two O atoms [Cu\u2014O = 1.908\u2005(4)\u2005\u00c5] from the tetra\u00addentate Schiff base ligand and by one O atom [Cu\u2014O = 2.324\u2005(6)\u2005\u00c5] of the methanol molecule in a square-pyramidal geometry. In the crystal structure, inter\u00admolecular O\u2014H\u22efO hydrogen bonds link complex mol\u00adecules into extended chains along [001].In the title compound, [Cu(C DOI: 10.1107/S1600536809011179/lh2794Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II compound, [Ni(C8H5O3)2(C10H14N2O)2(H2O)2], the central NiII atom is coordinated by two O atoms from two 4-formyl\u00adbenzoate (FOB) ligands, two O atoms from two water mol\u00adecules and two N atoms from two diethyl\u00adnicotinamide (DENA) ligands. The coordination geometry is slightly distorted octa\u00adhedral, with four O atoms in the equatorial plane and two N atoms in axial positions. Intra\u00admolecular O\u2014H\u22efO hydrogen bonds are observed. In the crystal structure, mol\u00adecules are linked into chains along the a axis by inter\u00admolecular O\u2014H\u22efO hydrogen bonds. The structure is further stabilized by \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings of DENA units, with a centroid\u2013centroid distance of 3.668\u2005(2)\u2005\u00c5.In the title centrosymmetric mononuclear Ni DOI: 10.1107/S1600536809006345/ci2769Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "An uncoordinated dmphen and an uncoordinated water mol\u00adecule cocrystallized with each complex mol\u00adecule. Intra- and inter\u00admolecular O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds are also present between the coordinated 3-HBA and water mol\u00adecules and the uncoordinated dmphen and water mol\u00adecules in the crystal. The packing of the structure is further stabilized by \u03c0\u2013\u03c0 stacking inter\u00adactions involving dmphen mol\u00adecules, with a centroid\u2013centroid separation of 3.705\u2005(3)\u2005\u00c5.In the title compound, [Mn(C DOI: 10.1107/S1600536809025926/pv2157Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two TbIII ions are linked by four bridging benzoate ions, with a Tb\u22efTb distance of 3.9280\u2005(6)\u2005\u00c5. Additionally, each TbIII ion is coordinated by one phenanthroline heterocycle and a bidentate benzoate ion. The irregular nine-coordinated geometry of the TbIII ion is composed of seven O and two N atoms. The mol\u00adecular structure is stabilized by intra\u00admolecular C\u2014H\u22efO hydrogen bonds. In the crystal structure, mol\u00adecules are linked into chains along the a axis by inter\u00admolecular C\u2014H\u22efO hydrogen bonds. The crystal structure is further stabilized by inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. Weak \u03c0\u2013\u03c0 inter\u00adactions are also observed [centroid\u2013centroid distances = 3.6275\u2005(14)\u20133.6604\u2005(14)\u2005\u00c5].The asymmetric unit of the title complex, [Tb For rel al. 1999, 2002 \u25b6;2(C7H5O2)6(C12H8N2)2]\u00b72C7H6O2 = 0.018 wR(F 2) = 0.101 S = 1.20 11944 reflections460 parametersH-atom parameters constrainedmax = 1.41 e \u00c5\u22123 \u0394\u03c1min = \u22122.25 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXTL (Sheldrick, 2008SHELXTL; molecular graphics: SHELXTL; software used to prepare material for publication: SHELXTL and PLATON (Spek, 2009Data collection: 10.1107/S1600536810016788/rz2444sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536810016788/rz2444Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II ion in the title complex, [HgCl2(C13H12N2)], adopts a distorted tetra\u00adhedral geometry being coordinated by two Cl anions and by two N atoms of the benz\u00adyl(2-pyridyl\u00admethyl\u00adene)amine ligand. The Cl\u2014Hg\u2014Cl plane is twisted at 70.1\u2005(1)\u00b0 from the mean plane of the chelate ring. In the crystal structure, inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.793\u2005(3)\u2005\u00c5] between the aromatic rings link the mol\u00adecules into zigzag chains extending along [010].The Hg DOI: 10.1107/S1600536810035889/cv2760Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One of PMB ions acts as a bidentate ligand while the other and the two INA are monodentate ligands. An O\u2014H\u22efO hydrogen bond links the uncoordinated water mol\u00adecule to the carboxyl groups of the complex. The dihedral angles between the carboxyl groups and the adjacent benzene rings are 10.28\u2005(11) and 21.24\u2005(9)\u00b0, while the two benzene rings and the two pyridine rings are oriented at dihedral angles of 6.90\u2005(4) and 88.64\u2005(4)\u00b0, respectively. In the crystal structure, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a supra\u00admolecular structure. A \u03c0\u2013\u03c0 contact between the benzene rings [centroid\u2013centroid distance = 3.911\u2005(1)\u2005\u00c5] may further stabilize the crystal structure. Weak C\u2014H\u22ef\u03c0 inter\u00adactions involving the pyridine rings also occur in the crystal structure.In the crystal structure of the title compound, [Cd(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)]\u00b7H2O = 0.020 wR(F 2) = 0.053 S = 1.14 7154 reflections404 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.56 e \u00c5\u22123 \u0394\u03c1min = \u22120.38 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S1600536810008366/xu2732sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810008366/xu2732Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The xanthene ring system is essentially planar, with an average deviation of 0.020\u2005\u00c5, and the substituent benzene ring forms a dihedral angle of 85.89\u2005(2)\u00b0 with it. The Sn\u2014Cl distances are in the range 2.4237\u2005(3)\u20132.4454\u2005(3)\u2005\u00c5. There are N\u2014H\u22efCl hydrogen bonds between SnCl6 2\u2212 ions and rhodamine 6G cations as well as \u03c0\u2013\u03c0 stacking inter\u00adactions between rhodamine 6G cations (inter\u00adplanar distance of 3.827\u2005\u00c5).In the title compound, bis({6-ethylamino-10-[2-(methoxycarbonyl)phenyl]-2,7-dimethylxanthen-3-ylidene}ethanaminium) hexachloridotin(IV) acetonitrile disolvate, (C DOI: 10.1107/S1600536807066287/pv2056Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Sn atom is coordinated by one N [Sn\u2014N = 2.187\u2005(3)\u2005\u00c5], two O [Sn\u2014O = 2.123\u2005(3) and 2.174\u2005(3)\u2005\u00c5] and two C [Sn\u2014C = 2.096\u2005(4) and 2.101\u2005(4)\u2005\u00c5] atoms in a distorted trigonal-bipyramidal geometry. The crystal packing exhibits weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds, which link the mol\u00adecules into centrosymmetric dimers with an Sn\u22efSn separation of 4.330\u2005(6)\u2005\u00c5, and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance of 3.690\u2005(5)\u2005\u00c5 between the benzene rings of neighbouring mol\u00adecules].In the title mol\u00adecule, [Sn(CH DOI: 10.1107/S1600536808040786/cv2482Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular \u03c0\u2013\u03c0 stacking between parallel pyridine rings [face-to-face distance 3.32\u2005(4)\u2005\u00c5] and C\u2014H\u22efBr and C\u2014H\u22efO hydrogen-bonding interactions are present in the crystal structure.In the mol\u00adecule of the title compound, [ZnBr DOI: 10.1107/S1600536810017551/xu2761Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each AgI ion is coordinated by two N atoms from two different isonicotinate ligands. The crystal structure exhibits a two-dimensional heterometallic polymeric layer. O\u2014H\u22efO hydrogen bonds involving the coordinated and uncoordinated water mol\u00adecules and intra\u00adlayer \u03c0\u2013\u03c0 inter\u00adactions between the pyridine rings [centroid\u2013centroid distances = 3.571\u2005(2) and 3.569\u2005(2)\u2005\u00c5] are observed. Each layer inter\u00adacts with two neighboring ones via Ag\u22efO(H2O) contacts and inter\u00adlayer \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.479\u2005(3) to 3.530\u2005(3)\u2005\u00c5], leading to a three-dimensional supra\u00admolecular network.In the title complex, {[Ag DOI: 10.1107/S1600536809042342/hy2237Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Ni(C8H7O2)2(C6H6N2O)2(H2O)2], is centrosymmetric with the Ni atom located on an inversion center. The mol\u00adecule contains two 4-methyl\u00adbenzoate (PMB) and two nicotinamide (NA) ligands and two coordinated water mol\u00adecules, all ligands being monodentate. The four O atoms in the equatorial plane around the Ni atom form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 26.15\u2005(10)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 87.81\u2005(4)\u00b0. In the crystal structure, inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network. The \u03c0\u2013\u03c0 contact between the benzene rings [centroid\u2013centroid distance = 3.896\u2005(1)\u2005\u00c5] may further stabilize the crystal structure. A weak C\u2014H\u22ef\u03c0 inter\u00adaction involving the pyridine ring also occurs.The title Ni N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.030 wR(F 2) = 0.068 S = 1.05 3390 reflections204 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.48 e \u00c5\u22123 \u0394\u03c1min = \u22120.50 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S1600536810007385/xu2730sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810007385/xu2730Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One of the Br atoms is disordered over two sites [site-occupancy factors = 0.51\u2005(6) and 0.49\u2005(6)]. The N\u2014C\u2014N angles in the cations are 110.7\u2005(6) and 111.4\u2005(7)\u00b0. In the crystal packing, a supra\u00admolecular chain is formed via both weak inter\u00admolecular C\u2014H\u22efBr hydrogen bonds and \u03c0\u2013\u03c0 aromatic ring stacking inter\u00adactions [centroid\u2013centroid separation = 3.803\u2005(1)\u2005\u00c5].In the title compound, (C DOI: 10.1107/S1600536809047461/zs2017Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The compound consists of an anionic complex, a doubly protonated propane-1,3-diamine as a counter-ion and 3.5 uncoord\u00adinated water mol\u00adecules. The coordination polyhedron around the ZnII atom is distorted octa\u00adhedral, defined by four O atoms and two N atoms from two Hchel ligands. In the crystal structure, O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds along with \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.473\u2005(2)\u2005\u00c5] are observed to reinforce the crystal cohesion.The asymmetric unit of the title compound, (C DOI: 10.1107/S1600536809041634/hy2228Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Two intra\u00admolecular O\u2014H\u22efO hydrogen bonds stabilize the conformation of the anion. The polymeric three-dimensional supra\u00admolecular architecture is formed via coordination inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions involving centrosymmetrically related pyrone rings, with a centroid\u2013centroid separation of 3.513\u2005(2)\u2005\u00c5.In the polymeric title compound, [K(C DOI: 10.1107/S1600536808041779/rz2268Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "This unit coordinates to a CaII cation in an O,O\u2032,O\u2032\u2032-tridentate fashion, generating a distorted CaCo3O4 cubane-type cluster. The Ca\u2014\u03bc3-O distances [2.429\u2005(5)\u20132.572\u2005(6)\u2005\u00c5] are much longer than the Co\u2014\u03bc3\u2014O bonds [1.895\u2005(5)\u20131.941\u2005(5)\u2005\u00c5]. The CaII cation is also coord\u00adinated by five water mol\u00adecules with Ca\u2014O distances in the range 2.355\u2005(6)\u20132.543\u2005(6)\u2005\u00c5. There are three additional uncoordinated water mol\u00adecules in the asymmetric unit, the occupancy of which refined to 0.54\u2005(3). In H2O (or D2O), the title complex hydrolyses to Ca2+ aq cations and [Co3(ida)3(\u03bc2-OH)3(\u03bc3-O)]2\u2212 anions. In the title compound, [CaCo DOI: 10.1107/S1600536810010998/sj2753Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ZnII ion is located on a crystallographic twofold rotation axis and assumes a distorted tetra\u00adhedral ZnN2O2 coordination geometry. Mol\u00adecules are linked by an inter\u00admolecular O\u2014H\u22efO hydrogen bond and \u03c0\u2013\u03c0 stacking inter\u00adactions between pyridine rings [centroid\u2013centroid speparation 3.594\u2005(1)\u2005\u00c5].The title complex, [Zn(C DOI: 10.1107/S1600536809027147/bx2223Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In ligand L, the imidazole and triazole rings form a dihedral angle of 74.25\u2005(8)\u00b0. Mol\u00adecules are assembled into a three-dimensional structure via inter\u00admolecular O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efN hydrogen-bonds, and \u03c0\u2013\u03c0 inter\u00adactions with a short distance of 3.665\u2005(2)\u2005\u00c5 between the centroids of the imidazole and triazole rings of neighbouring mol\u00adecules.In the title compound, [Mn(C DOI: 10.1107/S1600536810012626/cv2709Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The tridentate function of each ligand is completed by two N atoms coordinated to the two CuII atoms [Cu\u2014N = 1.933\u2005(2)\u2005\u00c5]. The separation distance of two CuII atoms in a cluster is 2.988\u2005(1)\u2005\u00c5. The dihedral angle between the six-membered chelate rings is 2.13\u2005(9)\u00b0. The perchlorate counter-anion is disordered over two sites in a 0.58\u2005(10):0.42\u2005(10) ratio.The title complex, [Cu DOI: 10.1107/S1600536810008512/kp2237Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The \u2013OH substituent only results in weak C\u2014H\u22efO weak inter\u00adactions between one of cyclo\u00adpenta\u00addienyl (Cp) ring H atoms and the O atom of a neighbouring mol\u00adecule with a distance of 3.308\u2005(3)\u2005\u00c5 between the donor and acceptor atoms. The inter\u00adplanar angle between the Cp and benzene rings is 13.0\u2005(4)\u00b0. There are also weak O\u2014H\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions involving the unsubstituted Cp and the benzene ring, respectively.The title compound, [Fe(C DOI: 10.1107/S1600536808039524/dn2401Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angles between meso-substituted chloro\u00adphenyl rings and the basic tetra\u00adpyrrolic ring are 82.66\u2005(9), 82.16\u2005(7), 83.97\u2005(11) and 76.87\u2005(8)\u00b0. In one of the phosphine ligands, the two terminal methyl groups are disordered over two positions with refined site-occupancy ratios of 0.70\u2005(7):0.30\u2005(7) and 0.66\u2005(2):0.34\u2005(2). In the crystal structure, mol\u00adecules are linked together along the a axis by inter\u00admolecular C\u2014H\u22efCl inter\u00adactions. The crystal structure is further stabilized by intra\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN inter\u00adactions and inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions.In the mol\u00adecule of the title compound, [Co(C DOI: 10.1107/S1600536809019163/at2788Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, there is a \u03c0\u2013\u03c0 stacking inter\u00adaction involving a pyrazole ring and a symmetry-related pyridine ring with a centroid\u2013centroid distance of 3.578\u2005(3)\u2005\u00c5.In the title complex, [Cd(NCS) DOI: 10.1107/S1600536809039920/lh2919Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "A twofold rotation axis passes through the Mg atom. In the crystal structure, the cations and anions are linked by inter\u00admolecular O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.804\u2005(2)\u2005\u00c5] into layers parallel to the ac plane.In the mononuclear title complex, [Mg(C DOI: 10.1107/S1600536808022150/im2068Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The torsion angles for the Te\u2014Re\u2014Te\u2014Re sequence of atoms are 19.29\u2005(18) and 16.54\u2005(16)\u00b0 in the two mol\u00adecules. Thus, the Re\u2014Te four-membered rings in the two mol\u00adecules deviate significantly from planarity. Two intra\u00admolecular C\u2014H\u22efO inter\u00adactions occur in one of the mol\u00adecules. Te\u2014Te [4.0551\u2005(10)\u2005\u00c5] inter\u00adactions between the two mol\u00adecules and weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions stabilize the crystal packing.The title compound, [Re DOI: 10.1107/S1600536810014297/sj2772Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angle between the two phenyl rings is 48.14\u2005(6)\u00b0. In the crystal structure, inter\u00admolecular N\u2014H\u22efO hydrogen bonds link mol\u00adecules into centrosymmetric dimers. These dimers are, in turn, linked into a two-dimensional network via weak N\u2014H\u22ef\u03c0(arene) inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions with centroid\u2013centroid distances of 3.6937\u2005(11)\u2005\u00c5.The crystal structure determination of the title compound, C DOI: 10.1107/S1600536809046133/sj2668Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "N2OS and N2O2, each exhibiting a distorted square-planar geometry. \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings of neighbouring complexes [centroid\u2013centroid distance = 3.856\u2005(5)\u2005\u00c5] link pairs of mol\u00adecules into centrosymmetric dimers, which are further packed into stacks along the b axis with relatively short Cu\u22efCu separations of 3.482\u2005(1)\u2005\u00c5. Weak inter\u00admolecular C\u2014H\u22efN hydrogen bonds help to consolidate the crystal packing.In the title dinuclear complex, [Cu DOI: 10.1107/S1600536809040951/cv2616Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each independent [FeIII(mnt)2]\u2212 unit forms a centrosymmetric dimer supported by two inter\u00admonomer FeIII\u2014S bonds [Fe\u2014S = 2.4715\u2005(9) and 2.4452\u2005(9)\u2005\u00c5]. In the crystal structure, the dimers form one-dimensional \u03c0\u2013\u03c0 stacks along the a axis, with an inter\u00adplanar separation of 3.38\u2005(6)\u2005\u00c5.The title compound, [Fe(C DOI: 10.1107/S1600536808036805/lh2727Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds and weak \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance 3.501\u2005(3)\u2005\u00c5]. As well as the cation, two anions and two dimethyl\u00adformamide solvent mol\u00adecules, the asymmetric unit also contains an ethanol solvent molecule with 0.25 occupancy.In the title compound, [Ni(C DOI: 10.1107/S1600536809021163/lh2826Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII atom is coordinated by two pydc dianions acting as tridentate ligands, and forming five-membered chelate rings with copper(II) as the central atom. The CuII atom is surrounded by four O atoms in the equatorial plane and two pyridine N atoms in axial positions, resulting in a distorted octa\u00adhedral coordination geometry. In the crystal, there are two types of O\u2014H\u22efO and N\u2014H\u22efO hydrogen-bonding synthons linking the anionic and cationic fragments and the water mol\u00adecules, namely R 4 4(16), and R 4 2(8). There are also weak C\u2014H\u22efO hydrogen bonds, \u03c0\u2013\u03c0 stacking inter\u00adactions [the shortest centroid\u2013centroid distance is 3.350\u2005(2)\u2005\u00c5], and a C\u2014O\u22ef\u03c0 inter\u00adaction [O\u22efcentroid distance = 3.564\u2005(2)\u2005\u00c5], which connect the various components into a three-dimensional network.The asymmetric unit of the title compound, (C DOI: 10.1107/S160053681003059X/vm2038Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CoII atoms are bridged by the PZDC dianions, forming an infinite one-dimensional chain running along the b axis. Adjacent chains pack together through \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid separations = 3.498\u2005(4) and 3.528\u2005(4)\u2005\u00c5], and O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds involving the water mol\u00adecule complete the structure.In the title compound, [Co(C DOI: 10.1107/S1600536808027177/hb2775Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The most important bond distances include Pd\u2014P of 2.380\u2005(2)\u2005\u00c5 and Pd\u2014Br of 2.515\u2005(2)\u2005\u00c5. Weak inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of adjacent mol\u00adecules [centroid\u2013centroid distance = 3.949\u2005(6)\u2005\u00c5] are present via crystallographic inversion centres, resulting in a one-dimensional supra\u00admolecular architecture.The title compound, [PdBr DOI: 10.1107/S1600536808030845/si2114Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Bi\u2014S and S\u2014P bond lengths are in the ranges 2.7694\u2005(18)\u20132.8391\u2005(17) and 2.019\u2005(2)\u20132.035\u2005(2)\u2005\u00c5, respectively. The crystal structure is consolidated by C\u2014H\u22efS hydrogen bonds. Intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions also play a role in stabilizing the mol\u00adecules.The title compound, [Bi(C DOI: 10.1107/S1600536810015618/wm2334Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Cu2Cl4(C24H21N5O4)2], was synthesized by the reaction of CuCl2\u00b72H2O with the tripodal ligand 2,2\u2032-diphthalimide (L). Each of the CuII ions is coordinated by two N atoms from the ligand, two bridging Cl atoms and one terminal Cl atom. The CuII coordination can be best be described as a transition state between four- and five-coordination, since one of the bridging Cl atoms has a much longer Cu\u2014Cl bond distance [2.7069\u2005(13)\u2005\u00c5] than the other [2.2630\u2005(12)\u2005\u00c5]. In addition, the Cu\u22efCu distance is 3.622\u2005(1)\u2005\u00c5. The three-dimensional structrure is generated by N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.658\u2005(4) and 4.020\u2005(4)\u2005\u00c5].The centrosymmetric dinuclear Cu DOI: 10.1107/S1600536809045565/kp2236Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Cu2+ and the Cl\u2212 ion are imposed by a twofold rotation axiss which also bisects the equally disordered benzoate anion. In the crystal, the mol\u00adecules are assembled into chains along [010] by C\u2014H\u22efCl, O\u2014H\u22efCl and C\u2014H\u22efO hydrogen-bonding inter\u00adactions. The resulting chains are further connected into two-dimensional supra\u00admolecular layers parallel to [100] by inter\u00adchain \u03c0\u22ef\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.823\u2005(5)\u2005\u00c5] between the phenanthroline ligands and the benzoic acid mol\u00adecules, and by C\u2014H\u22efO hydrogen-bonding inter\u00adactions. Strong \u03c0\u22ef\u03c0 stacking inter\u00adactions between adjacent phenantroline ligands [3.548\u2005(4)\u2005\u00c5] assemble the layers into a three-dimensional supra\u00admolecular architecture.In the title complex, [Cu(C DOI: 10.1107/S1600536810011487/zq2032Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The phenyl ring is nearly perpendicular to the plane of the tricyclic naphthofuran system [81.77\u2005(6)\u00b0] and is tilted slightly towards it. The 4-bromo\u00adphenyl ring is rotated out of the naphthofuran plane by a dihedral angle of 31.12\u2005(4)\u00b0. In the crystal structure, non-classical inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efBr hydrogen bonds are observed. The crystal structure also exhibits aromatic \u03c0\u2013\u03c0 inter\u00adactions between the furan ring and the central benzene ring of the adjacent naphthofuran system [centroid\u2013centroid distance = 3.768\u2005(3)\u2005\u00c5]. In addition, inter\u00admolecular C\u2014Br\u22ef\u03c0 inter\u00adactions [3.866\u2005(2)\u2005\u00c5] between the Br atom and the phenyl ring of the phenyl\u00adsulfinyl substituent are present.In the title compound, C DOI: 10.1107/S1600536809029250/im2130Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each Ag atom is connected by one bridging 4,4\u2032-bipyridine [Ag\u2014N = 2.177\u2005(4)\u00c5] and a terminal dicyanamide [Ag\u2014N = 2.108\u2005(4)\u2005\u00c5]. The Ag\u2014Ag interactions play a key role in constructing a unique neutral polymeric chain.In the title compound, [Ag DOI: 10.1107/S1600536809050491/pv2240Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal structure, inter\u00admolecular N\u2014H\u22efCl hydrogen bonds link the mol\u00adecules into centrosymmetric dimers. There is a \u03c0\u2013\u03c0 contact between the pyridine rings [centroid\u2013centroid distance = 3.896\u2005(5)\u2005\u00c5].In the mol\u00adecule of the title compound, [HgCl DOI: 10.1107/S1600536808040294/hk2588Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions between a methyl H atom of the methyl\u00adsulfinyl group and the benzene ring of the benzofuran system, and by an I\u22efO halogen bond of 3.173\u2005(3)\u2005\u00c5 and a nearly linear C\u2014I\u22efO angle of 171.7\u2005(1)\u00b0. In addition, the crystal structure exhibits weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds. The O atom of the carbonyl group and the butyl chain are both disordered over two positions with site-occupancy factors from refinement of 0.55\u2005(4) and 0.45\u2005(4) (for the O atom), and 0.76\u2005(2) and 0.24\u2005(2) (for the butyl group).In the title compound, C DOI: 10.1107/S1600536809000208/hg2458Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, inter\u00admolecular O\u2014H\u22efO hydrogen bonds link two CuL mol\u00adecules and four solvent mol\u00adecules into a centrosymmetric cluster. The crystal packing exhibits short inter\u00admolecular C\u22efC contacts of 3.185\u2005(4) and 3.232\u2005(4)\u2005\u00c5.In the title compound, [Cu(C DOI: 10.1107/S1600536809053720/cv2670Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure exhibits inter\u00admolecular O\u2014H\u22efO hydrogen bonds, short Cl\u22efCl contacts [3.334\u2005(1)\u2005\u00c5] and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance 3.621\u2005(11)\u2005\u00c5].In the title complex, [Cu(C DOI: 10.1107/S1600536808000044/cv2370Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The values of the bond angles at the Zn atom are in the range 99.4\u2005(1) to 113.2\u2005(1)\u00b0. The presence of O\u2014H\u22efO and O\u2014H\u22efBr intra\u00admolecular hydrogen bonds can explain the difference between the two Zn\u2014O [1.961\u2005(3)/2.015\u2005(3)\u2005\u00c5] and the two Zn\u2014Br [2.350\u2005(1)/2.378\u2005(1)\u2005\u00c5] bond lengths. The crystal structure is governed by C\u2014H\u22efO, C\u2014H\u22efBr and Zn\u2014Br\u22efCg(\u03c0-ring) inter\u00adactions.In the title compound, [ZnBr DOI: 10.1107/S1600536808016838/dn2351Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The oxycarbene group is nearly planar, with the phenyl ring twisted by an angle of 39.1\u2005(2)\u00b0 with respect to this plane. One of the cyclo\u00adpenta\u00addienyl rings undergoes an offset face-to-face \u03c0\u2013\u03c0 inter\u00adaction [3.544\u2005(6)\u2005\u00c5] with the symmetry-related cyclo\u00adpenta\u00addienyl ring of a neighbouring mol\u00adecule.The title compound, [TiW(C DOI: 10.1107/S1600536808036465/at2672Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two cyclo\u00adpenta\u00addienyl (Cp) rings are parallel to each other in both mol\u00adecules, forming dihedral angles of 2.3\u2005(3) and 1.0\u2005(3)\u00b0, respectively, and adopt an eclipsed conformation. The mean plane of the semicarbazone group is twisted slightly away from the attached Cp ring in both mol\u00adecules, the dihedral angles between the mean plane and the Cp ring being 15.3\u2005(2) and 10.8\u2005(2)\u00b0. The ethyl group in mol\u00adecule A is coplanar with the mean plane of the semicarbazone group [C\u2014N\u2014C\u2014C torsion angle = \u2212175.2\u2005(4)\u00b0], whereas it is nearly perpendicular in mol\u00adecule B [C\u2014N\u2014C\u2014C torsion angle = 84.8\u2005(6)\u00b0]. In the crystal structure, inter\u00admolecular N\u2014H\u22efS hydrogen bonds link the mol\u00adecules into dimers. These dimers are further linked into chains via inter\u00admolecular C\u2014H\u22efS hydrogen bonds. The crystal studied was a non-merohedral twin with a refined ratio of the twin components of 0.265\u2005(2):0.735\u2005(2).The asymmetric unit of title compound, [Fe(C DOI: 10.1107/S1600536810018209/rz2450Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The [Fe(H2O)6]2+ octa\u00adhedron has point symmetry 3(CO)3]\u2212 anion has point symmetry 1 and mean bond lengths of Fe\u2014Br = 2.455\u2005(5)\u2005\u00c5 and Fe\u2014C = 1.809\u2005(2)\u2005\u00c5. The cation and anion complexes are mutually linked via O\u2014H\u22efBr hydrogen bonds with O\u22efBr distances of 3.340\u2005(3) to 3.388\u2005(3)\u2005\u00c5.In the title compound, [Fe(H DOI: 10.1107/S1600536809036198/om2275Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Co(C9H10NO2)2(C10H14N2O)2(H2O)2], is centrosymmetric. It contains two dimethyl\u00adamino\u00adbenzoate (DMAB) and two diethyl\u00adnicotinamide (DENA) ligands and two water mol\u00adecules, all of them being monodentate. The four O atoms in the equatorial plane around the Co atom form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two pyridine N atoms of DENA ligands with the Co\u2014N distance of 2.1519\u2005(11)\u2005\u00c5 in the axial positions. The Co atom is displaced out of the least-squares plane of the carboxyl\u00adate group by \u22120.781\u2005(1)\u2005\u00c5. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 5.05\u2005(7)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 71.48\u2005(5)\u00b0. In the crystal structure, inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network. Two weak C\u2014H\u22ef\u03c0 inter\u00adactions are also present.The title Co DOI: 10.1107/S1600536809030980/xu2578Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Two pyridine-2,6-dicarboxyl\u00adate (pydc) ligands are bound to the NiII ion, giving an NiN2O4 bonded set. The coordination geometry around the NiII atom is distorted octa\u00adhedral. There are two types of robust O\u2014H\u22efO hydrogen-bond synthons, namely R 6 6(24) and R 2 4(8), which link the complex anions and water mol\u00adecules to each other. N\u2014H\u22efO hydrogen bonds connect the stacks of anions and cations in the structure. Other inter\u00admolecular inter\u00adactions, including weak C\u2014H\u22efO hydrogen bonds, \u03c0\u2013\u03c0 [shortest centroid\u2013centroid distance = 3.336\u2005(7)\u2005\u00c5] and C\u2014O\u22ef\u03c0 [O\u22efcentroid distance = 3.562\u2005(10)\u2005\u00c5] inter\u00adactions, connect the various components.The title compound, (C DOI: 10.1107/S1600536810016776/hy2305Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The average Hf\u2014O and Hf\u2014N distances are 2.096\u2005(3) and 2.398\u2005(3)\u2005\u00c5, respectively, and the average O\u2014Hf\u2014N bite angle is 70.99\u2005(11)\u00b0. The crystal packing is controlled by \u03c0\u2013\u03c0 inter\u00adactions between quinoline ligands of neighbouring mol\u00adecules and hydrogen-bonding inter\u00adactions. The inter\u00adplanar distances vary between 3.138\u2005(1) and 3.208\u2005(2)\u2005\u00c5, while the centroid\u2013centroid distances range from 3.576\u2005(1) to 4.074\u2005(1)\u2005\u00c5.In the title compound, [Hf(C DOI: 10.1107/S1600536809043244/bg2301Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Both RuIII atoms are octa\u00adhedrally coordinated by four chloride ligands in the equatorial plane and by two DMSO mol\u00adecules in apical positions within a RuCl4S2 donor set. The Na atom is surrounded by three chloride anions and three O atoms derived from three DMSO mol\u00adecules, with the resulting Cl3O3 donor set defining an octa\u00adhedron. The crystal structure is further stabilized by inter\u00adatomic inter\u00adactions of the types C\u22efCl [C\u2014Cl = 3.284\u2005(2)\u2005\u00c5], C\u2014H\u22efCl [C\u22efCl = 3.903\u2005(3)\u2005\u00c5] and C\u2014H\u22efO [C\u22efO = 3.376\u2005(3)\u2005\u00c5].The structure of the title compound, [NaRuCl DOI: 10.1107/S1600536810007063/tk2632Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The bpbda ligand, lying on an inversion center, bridges two CuI centres into a Z-shaped complex. Intra\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the dipyridophenazine ligand and the benzene ring of the bpbda ligand are observed [centroid\u2013centroid distance = 3.459\u2005(3)\u2005\u00c5]. The crystal structure also involves inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the dipyridophenazine ligands [centroid\u2013centroid distance = 3.506\u2005(3)\u2005\u00c5], which lead to a one-dimensional supra\u00admolecular structure.In the centrosymmetric dinuclear title compound, [Cu DOI: 10.1107/S1600536809031754/hy2208Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The distorted octa\u00adhedral environment of MnII is defined by two N atoms [Mn\u2014N = 2.217\u2005(4) and 2.132\u2005(5)\u2005\u00c5] and one O atom [Mn\u2014O 2.305\u2005(4)\u2005\u00c5]. There are inter\u00admolecular O\u2014H\u22efS hydrogen bonds and inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions between adjacent (2-pyrid\u00adyl)methano\u00adlate ligands [centroid\u2013centroid distance = 3.5569\u2005(7)\u2005\u00c5], leading to a chain structure running along [100].In the title complex, [Mn(NCS) DOI: 10.1107/S1600536810034483/bg2365Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure contains a CoII ion surrounded by the L 2\u2212 ligand in a slightly distorted square-planar fashion. Inter\u00admolecular C\u2014H\u22efO hydrogen-bonding contacts between the dichloro\u00admethane solvent mol\u00adecules and the meth\u00adoxy or carboxyl\u00adate O atoms are observed in the crystal structure. The planar complex mol\u00adecules stack through inversion related \u03c0\u2013\u03c0 inter\u00adactions between the six-membered rings of the vanillalimine half ligands. The distance between centroids is 3.498\u2005(2)\u2005\u00c5 and the perpendicular distance is 3.345\u2005\u00c5. A partial stacking is observed with a centroid\u2013centroid distance of 3.830\u2005(2)\u2005\u00c5, a perpendicular distance of 3.350\u2005\u00c5 and a slippage of 1.856\u2005\u00c5.The title compound, [Co(C DOI: 10.1107/S160053680900083X/si2147Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by aromatic \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of neighbouring mol\u00adecules [centroid\u2013centroid distance = 3.655\u2005(3)\u2005\u00c5] and by three inter\u00admolecular C\u2014H\u22efO non-classical hydrogen bonds.The title compound, C DOI: 10.1107/S1600536809008460/rk2133Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The metal atoms display a distorted tetragonal-pyramidal coordination geometry, and are linked by two \u03bc 2- and two 3\u03bc-hydroxo groups, assuming a chair-like conformation for the Cu4O2 core. In the crystal, the complex mol\u00adecules are linked into a three-dimensional network by inter\u00admolecular O\u2014H\u22efO, O\u2014H\u22efCl, C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions with centroid\u2013centroid separations of 3.724\u2005(2) and 3.767\u2005(3)\u2005\u00c5.The tetra\u00adnuclear copper(II) title complex, [Cu DOI: 10.1107/S160053680804381X/rz2280Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The structure displays O\u2014H\u22efO hydrogen bonding and inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions between 1,10-phenantroline ligands [inter\u00adplanar distance of 3.448\u2005(5)\u2005\u00c5]. In the structure of the title complex, [Cu(C DOI: 10.1107/S1600536809011659/gk2199Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Three chloridoaluminate groups and a hexa\u00admethyl\u00adbenzene mol\u00adecule are bound to the central lanthanum(III) ion, forming a distorted penta\u00adgonal bipyramid with the \u03b76-coordinated arene located at the apical position. The hexa\u00admethyl\u00adbenzene ligand disordered between two orientations in a 1:1 ratio is also involved in parallel-slipped \u03c0\u2013\u03c0 stacking inter\u00admolecular inter\u00adactions with a benzene solvent mol\u00adecule [centroid\u2013centroid distance 3.612\u2005(4)\u2005\u00c5].In the title compound, [Al DOI: 10.1107/S1600536809004899/cv2516Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, mol\u00adecules inter\u00adact via aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid separation = 3.6741\u2005(18)\u2005\u00c5].In the title compound, [Pt(CH DOI: 10.1107/S1600536809055470/hb5294Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, inter\u00admolecular O\u2014H\u22efCl hydrogen bonds link the mol\u00adecules into a chain structure along [010]. There is a \u03c0\u2013\u03c0 contact between the pyridine rings [centroid\u2013centroid distance = 3.824\u2005(5)\u2005\u00c5].The Ni atom in the title compound, [NiCl DOI: 10.1107/S1600536808043961/at2700Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The metal ion is coordinated via the thia\u00adzole nitro\u00adgen, imine nitro\u00adgen and thione sulfur atoms from each thio\u00adsemicarbazone ligand, and two coordinating units lie almost perpendicular to each other give dihedral angle = 81.89\u2005(1)\u00b0]. One thio\u00adsemicarbazone unit is found to bind a chloride anion through two hydrogen bonds, while the other is linked with the disordered crystal water molecule. Two mol\u00adecules are connected to each other through an inter\u00admolecular N\u2014H\u22efS inter\u00adaction, forming a centrosymmetric dimer. Dimers are linked into sheets by \u03c0\u2013\u03c0 stacking of two phenyl rings [shortest C\u22efC distance = 4.041\u2005(3)\u2005\u00c5].In the title compound, [Ni(C DOI: 10.1107/S1600536810013280/br2143Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The nitrate ion inter\u00adacts with the copper center [Cu1\u22efO3 = 2.579\u2005(4)\u2005\u00c5]. In the crystal, the cations, anions and water mol\u00adecules are linked by O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds.In the title mononuclear complex, [Cu(C DOI: 10.1107/S1600536809052350/zq2021Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The diethyl ether mol\u00adecule is located in a cavity provided by four O atoms of the ligand with weak C\u2014H\u22efO inter\u00adactions, generating two short O\u22efO contact distances [2.766\u2005(3) and 2.745\u2005(3)\u2005\u00c5] between the diethyl ether mol\u00adecule and the ligand. The crystal structure is stabilized by the weak C\u2014H\u22efO and C\u2014H\u22efN inter\u00adactions and \u03c0\u2013\u03c0 inter\u00adactions between the naphthyl ring system and the benzene ring [centroid\u2013centroid distance = 3.657\u2005(5)\u2005\u00c5] and between the two naphthyl ring systems [centroid\u2013centroid distance = 4.305\u2005(2)\u2005\u00c5].In the title complex, [Co(C For rel al. 1984; Marzill al. 1985; \u00c1lvarez al. 2002. For hyd al. 2002. For non al. 2002; De Ange al. 1996. For the al. 2008. 26H20N2O4)(NCS)]\u00b7C4H10O\u00b7CH2Cl2 = 0.075 wR(F 2) = 0.136 S = 1.21 7241 reflections397 parametersH-atom parameters constrainedmax = 0.69 e \u00c5\u22123 \u0394\u03c1min = \u22120.60 e \u00c5\u22123 \u0394\u03c1 CrystalClear (Rigaku, 2001CrystalClear; data reduction: CrystalStructure (Rigaku/MSC, 2007SIR97 (Altomare et al., 1999SHELXL97 (Sheldrick, 2008SHELXL97; software used to prepare material for publication: CrystalStructure.Data collection: 10.1107/S1600536809000841/is2372sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536809000841/is2372Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "\u00c5]. In the crystal, mol\u00adecules are connected into supra\u00admolecular chains via \u03c0\u2013\u03c0 inter\u00adactions formed by the pyridine rings [centroid\u2013centroid distance = 3.552\u2005(3)\u2005\u00c5] and these are connected into a two-dimensional array in the ac plane by C\u2014H\u22ef\u03c0 contacts. One of the tert-butyl groups is disordered over two orientations in a 0.734\u2005(6):0.266\u2005(6) ratio.The structure of the dinuclear title complex, [Cu al. 2006. Acta Cr DOI: 10.1107/S1600536810015060/hb5416Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The planar imidazole ring system [maximum deviation = 0.012\u2005(3)\u2005\u00c5] is oriented at a dihedral angle of 6.08\u2005(5)\u00b0 with respect to the protonated pyridine ring. An O\u2014H\u22efCl inter\u00adaction links the water mol\u00adecule to the dianion. In the crystal structure, inter\u00admolecular O\u2014H\u22efCl, N\u2014H\u22efO and N\u2014H\u22efCl inter\u00adactions link the mol\u00adecules into a three-dimensional network.In the title compound, (C DOI: 10.1107/S1600536809024568/hk2717Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There is a \u03c0\u2013\u03c0 contact between the pyridine rings [centroid\u2013centroid distance = 3.9758\u2005(5)\u2005\u00c5].In the mol\u00adecule of the title compound, [HgCl DOI: 10.1107/S1600536808032777/hk2551Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ZnII atom is bonded by two O atoms of the bidentate chelating sulfate ligand and four N atoms of the two chelating 2,2\u2032-bipyridine ligands. The Zn\u2014N bond distances range from 2.1287\u2005(17) to 2.1452\u2005(17)\u2005\u00c5 and the Zn\u2014O bond distance is 2.1811\u2005(15)\u2005\u00c5. The two chelating NCCN groups subtend a dihedral angle of 81.1\u2005(1)\u00b0. In the crystal structure, the [ZnSO4(C10H8N2)2] and C2H6O2 units are connected by inter\u00admolecular O\u2014H\u22efO hydrogen bonding, which leads to additional stabilization of the structure.The title compound, [Zn(SO DOI: 10.1107/S1600536809055433/bq2188Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, combined with weak (lone pair)\u22ef\u03c0 [O\u22efcentroid distance = 3.401\u2005(4)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking [centroid\u2013centroid distance = 3.975\u2005(3)\u2005\u00c5] inter\u00adactions, stabilize the three-dimensional supra\u00admolecular network.In the title compound, [Ag(NH DOI: 10.1107/S1600536810007725/hy2282Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two pyridine rings in each 2,2\u2032-bipyridine ligand form dihedral angles of 10.75\u2005(12) and 4.28\u2005(13)\u00b0. The crystal packing is stabilized by inter\u00adionic C\u2014H\u22efO hydrogen bonds, C\u2014H\u22ef\u03c0 inter\u00adactions and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions, with centroid\u2013centroid distances of 3.616\u2005(7)\u2005\u00c5.In the cation of the title compound, [CoCl(C DOI: 10.1107/S1600536809049034/rz2393Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The [Zn2Cl6(C8H4O4)]4\u2212 anions lie on centres of inversion and comprise two ZnCl3 groups bridged by benzene-1,4-dicarboxyl\u00adate. In addition to N\u2014H\u22efCl and N\u2014H\u22efO hydrogen bonds between the cations and anions, solvent water mol\u00adecules form O\u2014H\u22efO and O\u2014H\u22efCl hydrogen bonds to give a three-dimensional network.The title compound, (C DOI: 10.1107/S1600536808033011/bi2304Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The 2-pyridinium cations are linked to these chains through pyridinium\u2013water N\u2014H\u22efO hydrogen bonds and a two-dimensional network is formed through water bridges between sulfonate and 2-nitro O atoms, while the structure also has weak cation\u2013anion \u03c0\u2013\u03c0 aromatic ring inter\u00adactions [minimum ring centroid separation = 3.8441\u2005(13)\u2005\u00c5].In the structure of the title salt, 2C DOI: 10.1107/S1600536810014819/ng2763Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the indole ring system, the benzene and pyrrole rings are nearly coplanar, forming a dihedral angle of 1.95\u2005(8)\u00b0. The cyclo\u00adhexenone ring has an envelope conformation. In the crystal structure, pairs of strong N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into centrosymmetric dimers with R 2 2(10) ring motifs. \u03c0\u2013\u03c0 contacts between parallel pyrrole rings [centroid\u2013centroid distance = 3.776\u2005(2)\u2005\u00c5] may further stabilize the structure. A weak C\u2014H\u22ef\u03c0 inter\u00adaction is also observed.The title compound, C DOI: 10.1107/S160053680902385X/xu2543Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Both the stibine and arsine ligands are equatorial with respect to the Ru3 triangle. Additionally, each Ru atom carries one equatorial and two axial terminal carbonyl ligands. The three stibine-substituted phenyl rings make dihedral angles of 84.3\u2005(3), 80.4\u2005(3) and 70.5\u2005(3)\u00b0 with each other. The dihedral angles between the two phenyl rings are 85.9\u2005(3) and 75.2\u2005(3)\u00b0 for the two diphenyl\u00adarsine groups. In the crystal packing, mol\u00adecules are linked into chains down the c axis via inter\u00admolecular C\u2014H\u22efO hydrogen bonds. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions further stabilize the crystal structure.In the title DOI: 10.1107/S1600536809049927/sj2682Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Two terminal C atoms of one allyl group are disordered over two sites of equal occupancy. The crystal structure is stabilized by \u03c0\u2013\u03c0 stacking inter\u00adactions, the centroid\u2013centroid distance between nearly parallel [dihedral angle = 19.82\u2005(4)\u00b0] benzene and imidazole rings being 3.7885\u2005(15)\u2005\u00c5.In the title compound, C DOI: 10.1107/S1600536810007890/xu2721Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, inter\u00admolecular O\u2014H\u22ef and O\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid separation = 3.5535\u2005(13)\u2005\u00c5] link the mol\u00adecules into chains.In the title complex, [Mn(C DOI: 10.1107/S1600536809052659/hb5265Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "ZnN2O3 and ZnO5. All eight ZnII centers adopt a distorted square-pyramidal coordination; four ZnII ions have the N2O2 tetra\u00addentate Schiff base ligand bound in a basal plane and the coordinated water mol\u00adecule occupying the apical site, while the remaing four ZnII ions are bound by five O atoms from three Schiff base ligands. In the crystal structure, ZnII complex mol\u00adecules, coordinated and uncoord\u00adinated water mol\u00adecules and dimethyl sulfoxide mol\u00adecules are linked via O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional framework.The asymmetric unit of the title compound, tetra\u00adaqua\u00adtetrakis\u00ad{\u03bc II coordination complexes, see: Basak et al. 2007et al. 4(H2O)4]\u00b74C2H6OS\u00b72H2O = 0.050 wR(F 2) = 0.138 S = 1.02 5851 reflections314 parameters6 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.67 e \u00c5\u22123 \u0394\u03c1min = \u22121.34 e \u00c5\u22123 \u0394\u03c1 APEX2 (Bruker, 2005APEX2; data reduction: SAINT (Bruker, 2005SHELXTL (Sheldrick, 2008SHELXTL; molecular graphics: SHELXTL; software used to prepare material for publication: SHELXTL and PLATON (Spek, 2003Data collection: 10.1107/S1600536808017340/ci2611sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536808017340/ci2611Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit contains one-half of the whole mol\u00adecule, and there is an inversion center at the mid-point of the Cu\u22efCu bond. The octa\u00adhedral coordination of each Cu atom, with four O atoms in the equatorial plane, is completed by the N atom of the 6-methyl\u00adquinoline mol\u00adecule [Cu\u2014N = 2.212\u2005(2)\u2005\u00c5] and by another Cu atom [Cu\u22efCu = 2.6939\u2005(13)\u2005\u00c5]. The Cu atom lies 0.234\u2005\u00c5 out of the plane of the four O atoms. The molecular packing is stabilized by one intramolecular C\u2014H\u22efO as well as C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 interactions.In the title compound, [Cu DOI: 10.1107/S1600536808016516/bx2146Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angles between the central benzene ring and the two outer rings are 7.62\u2005(16) and 9.78\u2005(17)\u00b0. The crystal structure is stabilized by inter\u00admolecular C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions with a centroid\u2013centroid distance of 3.8218\u2005(19)\u2005\u00c5.In the title Schiff base complex, [Ni(C DOI: 10.1107/S1600536810035890/su2210Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "It is further extended to a two-dimensional layer structure by hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid-centroid distances of 3.560\u2005(2) and 3.666\u2005(2)\u2005\u00c5]. There is a C4 water chain in the structure whose repeat unit contains four water mol\u00adecules with O\u22efO distances in the range 2.748\u2005(3)\u20132.795\u2005(4)\u2005\u00c5. One of the two H atoms of each water of hydration is statistically distributed over two positions with equal occupancy.The title complex, {[Cd(C DOI: 10.1107/S1600536808037203/pv2111Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Hydrogen bonding O\u2014H\u22efO between water molecules and between water anions as well as \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances between phen rings = 3.38\u2005(7) and 3.40\u2005(5)\u2005\u00c5] are responsible for the supra\u00admolecular assembly.In the title compound, [Cu(CHO DOI: 10.1107/S1600536808035320/pk2125Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the anionic polymer, the Mn2+ ions are bridged by the pyridine\u00adcarboxyl\u00adate (pic) anion ligand, forming a chain structure along the c axis. The Mn2+ ion is six-coordinated in a distorted octa\u00adhedral environment by one N atom of the pyridine ring, two O atoms of the two carboxyl\u00adate groups, one O atom of the water mol\u00adecule and two Br atoms. The compound displays inter\u00admolecular N\u2014H\u22efO, N\u2014H\u22efBr, O\u2014H\u22efBr and O\u2014H\u22efO hydrogen bonding. There may also be inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between adjacent pyridine rings, with a centroid\u2013centroid distance of 3.992\u2005(4)\u2005\u00c5.The asymmetric unit of the title compound, {(C DOI: 10.1107/S1600536809016316/cs2117Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "These polymeric chains generate microchannels extending along [100], and these cavities are occupied by discrete tetra\u00addeca\u00admeric water clusters, which inter\u00adact with their surroundings and finally furnish the three-dimensional supra\u00admolecular network via 15 O\u2014H\u22efO, one O\u2014H\u22efS, two O\u2014H\u22efN and six N\u2014H\u22efO classical hydrogen bonds. 4,4-Bipyridine acts as an inserting component and hydrogen-bond acceptor, and it is a nonplanar mol\u00adecule with a dihedral angle of 33.12\u2005(13)\u00b0 between the pyridine rings. Owing to the numerous classical hydrogen bonds, the observed weak inter\u00admolecular C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions can be neglected with regard to stabilizing the network.In the title compound, {[La DOI: 10.1107/S1600536808031450/si2111Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There is a \u03c0\u2013\u03c0 stacking inter\u00adaction between the five-membered ring containing the Cu atom and a pyridine ring of a neighboring complex [centroid\u2013centroid distance = 3.567\u2005(2)\u2005\u00c5 and a perpendicular distance of 3.394\u2005\u00c5]. The crystal structure also contains inter\u00admolecular N\u2014H\u22efO, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, linking cations and anions. In addition, there is a short inter\u00admolecular contact [2.784\u2005(6)\u2005\u00c5] between an O atom of the coordinated nitrate group and its symmetry-related atom.In the title mononuclear complex, [Cu(NO DOI: 10.1107/S1600536808015523/wn2264Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atoms in the crystal structure of the title compound, (C2H10N2)[HgBr4]\u00b7H2O, are tetra\u00adhedrally coordinated by four Br atoms and the resulting [HgBr4]2\u2212 ions are inter\u00adconnected to the [NH3\u2014CH2\u2014CH2\u2014NH3]2+ ions and water mol\u00adecules by N\u2014H\u22efBr and O\u2014H\u22efBr bonds, forming a three-dimensional network. N\u2014H\u22efO inter\u00adactions are also present. The observed three different Hg\u2014Br distances of 2.5597\u2005(6), 2.6862\u2005(8) and 2.6923\u2005(8)\u2005\u00c5 in the tetra\u00adbromo\u00admercurate unit are due to the connection of Br atoms to different numbers of H atoms. The Hg, O and two Br atoms are located on a crystallographic mirror plane. The cation has The Hg DOI: 10.1107/S160053680902772X/bt5004Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The diimine ligand is equatorial and is bonded to the Re centre in an N,N\u2032-bidentate chelating fashion, with a bite angle of 77.7\u2005(2)\u00b0. The dihedral angle between the two benzene rings is 88.7\u2005(6)\u00b0. In the crystal structure, there are F\u22efO [2.856\u2005(9)\u2005\u00c5], Cl\u22efC [3.150\u2005(8)\u2005\u00c5] and O\u22efC [2.984\u2005(10)\u2005\u00c5] contacts which are shorter than the sum of the van der Waals radii for these atoms. In addition, symmetry-related mol\u00adecules are linked via inter\u00admolecular C\u2014H\u22efO, C\u2014H\u22efBr and the F\u22efO inter\u00adactions into one-dimensional chains extending along the a axis. The crystal structure is further stabilized by inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.571\u2005(5)\u2005\u00c5].In the title compound, [ReBr(C DOI: 10.1107/S1600536809001044/lh2753Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The compound is a seven-coordinate binuclear polymeric complex with distorted penta\u00adgonal bipyramidal geometry around CdII [Cd\u2014O = 2.247\u2005(4)\u20132.474\u2005(3)\u2005\u00c5]. In the binuclear monomeric units, the central atoms join together by O atoms of two bridging tridentate (hypydc)2\u2212 ligands, and the polymer propagates via two bridging water mol\u00adecules that link each CdII centre of one monomer to the adjacent neighbour. Propane-1,3-diamine (pn) does not appear in the product but plays a role as a base. Inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, and \u03c0\u2013\u03c0 stacking inter\u00adactions, with distances of 3.725\u2005(3) and 3.766\u2005(3)\u2005\u00c5, connect the various components.The title polymeric compound, {[Cd DOI: 10.1107/S1600536808030869/om2258Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CdII atom shows a distorted octa\u00adhedral environment, with four N atoms from two chelating 2,2\u2032-bipyridine ligands forming the equatorial plane and two sulfonate O atoms from two 9,10-dioxoanthracene-1,5-disulfonate anions occupying the apical positions. Weak C\u2014H\u22efO hydrogen-bonding contacts and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.6920\u2005(12) and 3.7095\u2005(12)\u2005\u00c5] connect the complex mol\u00adecules into a three-dimensional supra\u00admolecular framework.The title complex, [Cd(C DOI: 10.1107/S1600536809046881/si2219Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The NiII atom is coordinated by two N atoms [Ni\u2014N = 1.839\u2005(2)\u2005\u00c5] and two O atoms [Ni\u2014O = 1.8253\u2005(19)\u2005\u00c5] in an approximately square-planar geometry. In the crystal structure, inter\u00admolecular O\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a chain along [001].The monomeric title nickel(II) complex of a salicylaldimine, [Ni(C DOI: 10.1107/S160053680904063X/ds2006Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Cu\u22efCu separation is 2.5912\u2005(4)\u2005\u00c5. In the crystal, the mol\u00adecules are linked into polymeric chains propagating in [001] by inter\u00admolecular O\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, [Cu DOI: 10.1107/S1600536810013322/bq2206Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The conformation of the dimer is stabilized by inter\u00admolecular C\u2014H\u22efO hydrogen bonds and by \u03c0\u2013\u03c0 aromatic stacking inter\u00adactions involving the pyridine and benzene rings with centroid\u2013centroid separations of 3.624\u2005(3)\u2005\u00c5.The dinuclear title complex, [Cu DOI: 10.1107/S1600536809053665/rz2402Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CoII atom is coordinated by four O and two N atoms from two pydc ligands in a distorted octa\u00adhedral environment. The structure also contains three uncoordinated water mol\u00adecules. Extensive inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.565\u2005(14) and 3.425\u2005(14)\u2005\u00c5] and O\u22ef\u03c0 inter\u00adactions [O\u22efcentroid distance = 3.480\u2005(2)\u2005\u00c5] connect the various components in the crystal structure.The title compound, (C DOI: 10.1107/S1600536809021837/hy2201Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angle between the aromatic rings in the ligand is 8.70\u2005(16)\u00b0. The S atom is disordered over two positions in a 0.901\u2005(6):0.099\u2005(6) ratio. In the crystal, mol\u00adecules inter\u00adact by way of \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid separation = 3.720\u2005(2)\u2005\u00c5].In the title compound, [Cu(C DOI: 10.1107/S1600536810014212/hb5406Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Ni ion is coordinated by two N [Ni\u2014N = 1.8462\u2005(18)\u2005\u00c5] and two O [Ni\u2014O = 1.8645\u2005(14)\u2005\u00c5] atoms in a distorted square-planar geometry. The water mol\u00adecule and the Ni complex mol\u00adecule are paired via O\u2014H\u22efO hydrogen bonds.In the title compound, [Ni(C DOI: 10.1107/S1600536808039822/cv2487Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Short centroid\u2013centroid distances [3.762\u2005(5) and 3.867\u2005(5)\u2005\u00c5] between the aromatic rings of neighbouring cations suggest the existence of \u03c0\u2013\u03c0 inter\u00adactions. Inter\u00admolecular O\u2014H\u22efBr hydrogen bonds and weak C\u2014H\u22efO and C\u2014H\u22efBr inter\u00adactions consolidate the crystal packing.In the title compound, [CuBr(C DOI: 10.1107/S1600536809048995/cv2644Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Pairs of complex mol\u00adecules related by inversion centers are organized into dimers via pairs of Pb\u22efO inter\u00adactions [3.185\u2005(2)\u2005\u00c5] and stacking interactions between 2,2\u2032-bipyridine and 4-methyl\u00adbenzoate ligands, with a mean distance between their planes of 3.491\u2005\u00c5.In the title compound, [Pb(C DOI: 10.1107/S1600536810005544/gk2256Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The NiII atom lies on an inversion centre and is coordinated by four S atoms of O,O\u2032-diphenethyl dithio\u00adphosphate mol\u00adecules and two N atoms of 4-methyl\u00adpyridine mol\u00adecules. Important geometric data include Ni\u2014N = 2.100\u2005(3)\u2005\u00c5, and Ni\u2014S = 2.5101\u2005(10) and 2.4772\u2005(11)\u2005\u00c5.The title complex, [Ni(C DOI: 10.1107/S1600536808020898/dn2365Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "An offset face-to-face \u03c0\u2013\u03c0 stacking inter\u00adaction [centroid\u2013centroid distance = 3.674(4)\u2005\u00c5] and N\u2014H\u22efN and N\u2014H\u22efO hydrogen-bonding inter\u00adactions give rise to a one-dimensional supra\u00admolecular structure in the solid state.In the centrosymmetric title compound, [Ni(C DOI: 10.1107/S1600536808001116/pk2081Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One pair of phenyl rings attached to the two different P atoms are almost parallel, as are the other pair [dihedral angles = 8.7\u2005(5) and 8.9\u2005(5)\u00b0]. The planes of the two cyclo\u00adpenta\u00addienyl rings are inclined by 26.8\u2005(7)\u00b0 with respect to each other. The carbonyl groups at each Fe atom are almost perpendicular [C\u2014Fe\u2014C = 92.6\u2005(6) and 94.3\u2005(5)\u00b0].The title compound, [Fe DOI: 10.1107/S1600536808014931/bq2081Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordination geometry is distorted tetra\u00adhedral, with Sn\u2014C bond lengths in the range 2.133\u2005(6)\u20132.188\u2005(6)\u2005\u00c5 and with an Sn\u2014S bond length of 2.4516\u2005(19)\u2005\u00c5. The nonbonded S atom of the piperidine\u00addithio\u00adcarboxyl\u00adate anion makes an Sn\u22efS contact of 3.174\u2005(3)\u2005\u00c5.In the title compound, [Sn(C DOI: 10.1107/S1600536808021909/bi2291Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Ni atom lies on a crystallographic twofold rotation axis. The asymmetric unit contains one half-mol\u00adecule of the complex and a water mol\u00adecule. An inter\u00admolecular O\u2014H\u22efO hydrogen bond forms a four-membered ring, producing an R 1 2(4) ring motif involving a bifurcated hydrogen bond to the phenolate O atoms of the complex mol\u00adecule. In the crystal structure, mol\u00adecules are linked by \u03c0\u2013\u03c0 stacking inter\u00adactions, with centroid\u2013centroid distances in the range 3.5750\u2005(11)\u20133.7750\u2005(11)\u2005\u00c5. As a result of the twofold symmetry, the central benzene ring makes the same dihedral angle of 15.75\u2005(9)\u00b0 with the two outer benzene rings. The dihedral angle between the two hydroxy\u00adphenyl rings is 13.16\u2005(5)\u00b0. In the crystal structure, mol\u00adecules are linked into infinite one-dimensional chains by directed four-membered O\u2014H\u22efO\u2014H inter\u00adactions along the c axis and are further connected by C\u2014H\u22efO and \u03c0\u2013\u03c0 stacking into a three-dimensional network. An inter\u00adesting feature of the crystal structure is the short Ni\u22efO, O\u22efO and N\u22efN inter\u00adactions which are shorter than the sum of the van der Waals radii of the relevant atoms. The crystal structure is stabilized by inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and by \u03c0\u2013\u03c0 stacking inter\u00adactions.In the title complex, [Ni(C DOI: 10.1107/S1600536808026093/pk2114Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Weak inter\u00admolecular C\u2014H\u22efBr hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings [centroid\u2013centroid distances = 3.763\u2005(5) and 3.835\u2005(6)\u2005\u00c5] contribute to crystal-packing effects.In the title compound, [ZnBr DOI: 10.1107/S1600536810035658/jj2041Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The MgII ion is coordinated by four N atoms from two 1,10-phenanthroline ligands and two O atoms from coordinated water mol\u00adecules in a distorted octa\u00adhedral geometry. Inter\u00admolecular O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings [shortest centroid\u2013centroid separation = 3.527\u2005(2)\u2005\u00c5] link the cations, anions and 1,10-phenanthroline solvent mol\u00adecules into a hydrogen-bonded cluster.In the title compound, [Mg(C DOI: 10.1107/S1600536809030128/cv2597Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, the carboxyl groups are involved in inter\u00admolecular O\u2014H\u22efO hydrogen bonds, which link the mol\u00adecules into centrosymmetric dimers. These dimers are further packed into stacks along the c axis by weak C\u2014H\u22ef\u03c0 inter\u00adactions. In addition, the stacked mol\u00adecules exhibit a Br\u22efS inter\u00adaction of 3.4787\u2005(7)\u2005\u00c5.The title compound, C DOI: 10.1107/S1600536809005376/fj2195Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Intra\u00admolecular C\u2014H\u22efO inter\u00adactions occur. The mol\u00adecules are connected by inter\u00admolecular N\u2014H\u22efO, N\u2014H\u22efN and C\u2014H\u22efO hydrogen-bonding inter\u00adactions and C\u2014H\u22ef\u03c0 stacking inter\u00adactions, forming a three-dimensional supra\u00admolecular frameworkIn the title compound, [Sb(C DOI: 10.1107/S1600536809043578/gk2233Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the anion, four distorted [MoO6] octa\u00adhedra are connected via edge-sharing, forming an [Mo4O13]2\u2212 building block, composed of Mo\u2014O(t), Mo\u2014O(\u03bc2), Mo\u2014O(\u03bc3) and Mo\u2014O(\u03bc4) units, with Mo\u2014O distances ranging from 1.6858\u2005(15) to 2.4785\u2005(13)\u2005\u00c5. The \u03b3-type [Mo8O26]4\u2212 anion is completed by crystallographic inversion symmetry and is linked into an infinite chain along [100] by corner-sharing. The anionic chains and the cations are joined by N\u2014H\u22efO hydrogen bonds, generating layers extending parallel to (001).In the title compound, {(C DOI: 10.1107/S1600536810010111/wm2312Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two benzothia\u00adzole-2-thiol\u00adate ligands chelate via their thia\u00adzole N and thiol\u00adate S atoms while the methyl 2-(2-amino\u00adthia\u00adzol-4-yl)-2-(methoxy\u00adimino)acetate also acts as a chelate ligand binding through the thia\u00adzole and imino N atoms. Intra\u00admolecular N\u2014H\u22efN, C\u2014H\u22efN and C\u2014H\u22efO inter\u00adactions contribute to the mol\u00adecular conformation. In the crystal structure, inter\u00admolecular N\u2014H\u22efO hydrogen bonds produce R 1 2(6) rings and generate chains along the c axis. An extensive one-dimensional supra\u00admolecular network of N\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions is responsible for the crystal structure stabilization.In the title compound, [Ni(C DOI: 10.1107/S1600536810015072/sj2774Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The other Sn atom also has a distorted trigonal bipyramidal geometry, being coordinated by two O atoms of two carboxyl\u00adate anions, one bridging O atom and two butyl groups. An inter\u00adesting feature of the crystal structure is the short Sn\u22efO [2.756\u2005(4)\u2005\u00c5] and O\u22efO [2.608\u2005(3)\u2005\u00c5] inter\u00adactions. The \u2013BrCH2\u2014CHBr\u2013 segments of the two carboxyl\u00adate anions are disordered over two positions [site occupancies of 0.60\u2005(1)/0.40\u2005(1) and 0.53\u2005(2)/0.47\u2005(2)]. Weak non-directional C\u2014H\u22efO inter\u00adactions lead to the formation of infinte chains along the a axis; other weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions are also present.In the centrosymmetric tetra\u00adnuclear title complex, [Sn DOI: 10.1107/S1600536808037513/ng2514Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, aromatic \u03c0\u2013\u03c0 stacking occurs [centroid\u2013centroid distances 3.7630\u2005(14) and 3.7269\u2005(15)\u2005\u00c5], as well as an extensive O\u2014H\u22efO and C\u2014H\u22efO hydrogen-bonding networkIn the title compound, [Co(C DOI: 10.1107/S1600536808019843/hb2753Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In athymic rats, \u03b3/\u03b4 T-cells expressing intracellular \u03b2 proteinare present but at a lower frequency than in euthymic controls, suggesting that in the thymus,more \u03b3/\u03b4 T-cell precursors pass through a stage where functional \u03b2 rearrangement hasoccurred than in extrathymic sites. Analysis of TCR expression in purified transitory immatureCD4-8+ (iCD8SP) thymocytes and their spontaneously developing CD4+8+ (DP) progenyshowed that TCR\u03b3 mRNA is expressed in iCD8SP cells but not in their immediate DPprogeny that reinitiate RAG-1 transcription and commence \u03b1/\u03b2TCR expression. We concludethat rat \u03b3/\u03b4 T cells can separate from the \u03b1/\u03b2 lineage after TCR\u03b2 expression, but not afterentry into the DP compartment.The relationship between \u03b1/\u03b2 and \u03b3/\u03b4 T-cell lineages was studied in rats using RT-PCR analysisof TCR\u03b2 transcripts in \u03b3/\u03b4 T-cell hybridomas and an intracellular staining technique todetect TCR\u03b2 protein in primary \u03b3/\u03b4 T-cells. We report the presence of functional TCR\u03b2 transcriptsin 2/9 \u03b3/\u03b4 T-cell hybridomas. About 15 % of peripheral \u03b3/\u03b4 T-cells and thymocytes alsoexpress TCR\u03b2 protein, giving a minimum estimate for successful Tcrb rearrangement basedon"} +{"text": "The two CdII ions are connected by two tridentate bridging 2-hydroxy\u00adbenzoate anions. Each CdII ion is seven-coordinated by five O atoms from three 2-hydroxy\u00adbenzoate ligands and two N atoms from 1,10-phenanthroline. The 2-hydroxy\u00adbenzoate mol\u00adecules adopt two kinds of coordination mode, bidentate chelating and tridentate bridging\u2013chelating. Intra\u00admolecular hydrogen bonds between hydr\u00adoxy and carboxyl\u00adate groups from 2-hydroxy\u00adbenzoate groups and \u03c0\u2013\u03c0 stacking interactions between parallel 1,10-phenanthroline ligands [centroid\u2013centroid distances = 3.707\u2005(3) and 3.842\u2005(3)\u2005\u00c5] are observed. Furthermore, adjacent benzene rings from 2-hydroxy\u00adbenzoate ligands are involved in \u03c0\u2013\u03c0 inter\u00adactions with inter\u00adplanar distances of 3.642\u2005(3)\u2005\u00c5, thereby forming a chain along the a axis direction.The dinuclear title compound, [Cd DOI: 10.1107/S1600536808033886/zl2145Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each PtII center is coordinated by the amine groups of two (1-adamantylmeth\u00adyl)amine ligands and two Cl atoms in a cis-square-planar arrangement. The PtII centers lie slightly outside [0.031\u2005(4) and 0.038\u2005(4)\u2005\u00c5] the coordination planes. The N\u2014Pt\u2014N and Cl\u2014Pt\u2014Cl angles [92.1\u2005(4)\u201392.30\u2005(11)\u00b0] are slightly more open than the N\u2014Pt\u2014Cl angles [87.3\u2005(3)\u201388.3\u2005(3)\u00b0]. N\u2014H\u22efO and N\u2014H\u22efCl inter\u00admolecular hydrogen bonds are observed, forming two discrete pairs of complexes and solvent mol\u00adecules.The asymmetric unit of the title compound {systematic name: DOI: 10.1107/S1600536809037982/hy2221Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The carboxyl\u00adate groups are approximately coplanar with the thio\u00adphene ring [dihedral angle = 4.0\u2005(1)\u00b0] and the Sn\u2014O bond distance of 2.058\u2005(4)\u2005\u00c5 is comparable to that in related organotin carboxyl\u00adates.Mol\u00adecules of the title compound, [Sn DOI: 10.1107/S1600536809020273/bi2371Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The four O atoms in the equatorial plane around the Ni atom form a slightly distorted square-planar arrangement with an average Ni\u2014O bond length of 2.079\u2005\u00c5, and the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the nicotinamide (NA) ligands in the axial positions. The dihedral angle between the carboxyl group and the attached benzene ring is 28.28\u2005(11)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 8.31\u2005(4)\u00b0. In the crystal structure, O\u2014H\u22efO, N\u2014H\u22efO, C\u2014H\u22efO, and C\u2014H\u22efF hydrogen bonds link the mol\u00adecules into a three-dimensional network. \u03c0\u2013\u03c0 Contacts between the pyridine and benzene rings [centroid\u2013centroid distance = 3.626\u2005(1)\u2005\u00c5] may further stabilize the crystal structure. The 2-fluoro\u00adbenzoate anion is disordered over two orientations, with an occupancy ratio of 0.85:0.15.The asymmetric unit of the title complex, [Ni(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(H2O)4](C7H4FO2)2 = 0.026 wR(F 2) = 0.071 S = 1.04 3339 reflections221 parameters7 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.55 e \u00c5\u22123 \u0394\u03c1min = \u22120.70 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S1600536809040392/xu2610sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536809040392/xu2610Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Since assay constraints limited the data at lower doses, conventional analysis was not feasible and a \u2018population approach\u2019 was used. Bound concentrations were best described by a biexponential model and further analyses revealed a small influence of dose or weight on V1 but no identifiable effects of age, body surface area, renal or hepatic function. The final model was: clearance (Q) 0.194 l h\u22121; central compartment volume (V1) 4.48 \u00d7 (1+0.00074 \u00d7 dose (mg)) l; peripheral compartment volume (V2) 7.94 l; intercompartmental clearance 0.685 l h\u22121. Distribution and elimination half-lives had median estimates of 2.7 h and 49 h respectively. Free doxorubicin was present at most sampling times with concentrations around 1000 times lower than bound doxorubicin values. Data were best described using a biexponential model and the following parameters were estimated: apparent clearance 180 l h\u22121; apparent V1 (l) 1450 \u00d7 (1+0.0013 \u00d7 dose (mg)), apparent V2 (l) 21 300 \u00d7 (1\u20130.0013 \u00d7 dose (mg)) \u00d7 (1+2.95 \u00d7 height (m)) and apparent Q 6950 l h\u22121. Distribution and elimination half-lives were 0.13 h and 85 h respectively. \u00a9 1999 Cancer Research CampaignDoxorubicin pharmacokinetics were determined in 33 patients with solid tumours who received intravenous doses of 20\u2013320 mg m"} +{"text": "The structure contains [Mn(C10H8N2)(H2O)4]2+ cations with the MnII atoms lying on a centres of inversion and bridged into a linear chain along the a axis by 4,4\u2032-bipy ligands, surrounded by HL \u2212 anions and uncoordinated water mol\u00adecules. Extensive O\u2014H\u22efO hydrogen-bonding and weak \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.7572 \u2005(3)\u2005\u00c5] between the constituents lead to the formation of a three-dimensional supra\u00admolecular network.The title compound, {[Mn(C DOI: 10.1107/S1600536809028360/at2846Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Adjacent silver complexes are primarily linked together by Ag\u22efN inter\u00adactions, with Ag\u22efN separations of 2.877\u2005(2) and 3.314\u2005(2)\u2005\u00c5. On the other hand, one CF3SO3 \u2212 anion inter\u00adacts with hydrazone groups of two neighbouring ligands via N\u2014H\u22efO hydrogen bonds. These weak inter\u00admolecular inter\u00adactions contribute to the formation of supra\u00admolecular chains. In addition, there are Ag\u22efO inter\u00adactions [2.787\u2005(2)\u2005\u00c5] between Ag and O atoms from adjacent chains.In the title compound, [Ag(C DOI: 10.1107/S1600536809039579/bh2248Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atoms in the two independent mol\u00adecules of the title compound, [Cu(C7H5O3)2(C14H12N2)], are each coordinated by a bidentate 2,9-dimethyl-1,10-phenanthroline (dmphen) mol\u00adecule and two monodentate 2-hydroxy\u00adbenzoate anions in a distorted tetra\u00adhedral geometry. The crystal packing is stabilized by intra\u00admolecular hydrogen bonding and \u03c0\u2013\u03c0 inter\u00adactions between the dmphen rings of neighboring mol\u00adecules, with distances between their ring planes of 3.5670\u2005(4) and 3.5181\u2005(9)\u2005\u00c5.The Cu DOI: 10.1107/S1600536808036283/hg2438Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The L ligands are coordinated to the HoIII ions in three modes: chelating, bridging and bridging\u2013tridentate. Intra\u00admolecular C\u2014H\u22efO inter\u00adactions occur. The crystal packing is stabilized by inter\u00admolecular C\u2014H\u22efO inter\u00adactions and weak aromatic \u03c0\u2013\u03c0 inter\u00adactions between phen mol\u00adecules and the aromatic rings of the L ligands [centroid\u2013centroid distance = 3.821\u2005(2)\u2005\u00c5].In the centrosymmetric title compound, [Ho DOI: 10.1107/S1600536810036408/pv2299Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The PdII cation is situated on an inversion centre and exhibits an almost square-planar coordination by the S atoms of the TU ligands. The complex cations are connected through the thio\u00adcyanate ions via N\u2014H\u22efN [2.922\u2005(3)\u20133.056\u2005(3)\u2005\u00c5] and N\u2014H\u22efS [3.369\u2005(2)\u20133.645\u2005(2)\u2005\u00c5] hydrogen bonds.The title compound, [Pd(CH DOI: 10.1107/S160053680801088X/wm2176Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, the deca\u00advanadate [V10O28]6\u2212 anion 3}2(\u03bc-H2O)2]2+ units. Inter\u00adconnection of these aquasodium-ion-sandwiched deca\u00advanadates to chains parallel to [001] is effected by \u03bc-[{Na(H2O)3}2(\u03bc-H2O)2]2+ units, bridging adjacent deca\u00advanadates via O=V. The structure is consolidated by an extensive network of O\u2014H\u22efO hydrogen bonds.The title compound, {[Na(H DOI: 10.1107/S1600536810010251/br2141Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal packing exhibits inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions evidenced by relatively short distances [3.525\u2005(5)\u20133.937\u2005(6)\u2005\u00c5] between the centroids of the six-membered rings of neighbouring mol\u00adecules.In the title compound, [Cd(C DOI: 10.1107/S1600536807067918/cv2375Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Both the phosphine and arsine ligands are equatorial with respect to the Ru3 triangle. Additionally, each Ru atom carries one equatorial and two axial terminal carbonyl ligands. The three arsine-substituted phenyl rings make dihedral angles of 87.36\u2005(10), 81.96\u2005(10) and 73.37\u2005(11)\u00b0 with each other. The dihedral angles between the two phenyl rings are 88.08\u2005(12) and 80.15\u2005(10)\u00b0 for the two diphenyl\u00adphosphino groups. In the crystal packing, the mol\u00adecules are linked together as dimers via inter\u00admolecular C\u2014H\u22efO hydrogen bonds. These dimers are stacked down b axis. Inter\u00admolecular C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.6383\u2005(13)\u2005\u00c5] further stabilize the crystal structure.In the title DOI: 10.1107/S1600536809046704/sj2670Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "In the crystal structure, each RuII ion is coordinated by two Cl [Ru\u2014Cl = 2.4308\u2005(7) and 2.4139\u2005(7)\u2005\u00c5], two N [Ru\u2014N = 2.016\u2005(2) and 2.003\u2005(2)\u2005\u00c5], and two P [Ru\u2014P\u00a0= 2.3688\u2005(7) and 2.3887\u2005(7)\u2005\u00c5] atoms in a distorted octa\u00adhedral geometry. Packing inter\u00adactions include typical C\u2014H\u22ef\u03c0 contacts involving phenyl groups as well as weak hydrogen bonds between CH3CN methyl H atoms and Cl or solvent CH3CN N atoms.The title compound, [RuCl DOI: 10.1107/S1600536807065968/ci2537Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The tetra\u00adfluorido\u00adborate anions link the mononuclear cations through inter\u00admolecular C\u2014H\u22efF hydrogen-bonding inter\u00adactions, forming an infinite tape structure along [110]. Other weak inter\u00adactions occur: \u03c0\u2013\u03c0 stacking with centroid\u2013centroid distances of 3.820\u2005(2) and 3.898\u2005(1)\u2005\u00c5 between pyridyl rings and 3.610\u2005(2) and 3.926\u2005(2)\u2005\u00c5 between pyrazinyl rings as well as F\u22ef\u03c0 contacts involving the tetra\u00adfluorido\u00adborate anions and pyrazine rings [F\u22efcentroid = 2.999\u2005(3)\u2005\u00c5]; these combine with the hydrogen-bonding inter\u00adactions to link the mononuclear cations into a three-dimensional supra\u00admolecular architecture.In the title mononuclear complex, [Ag(C DOI: 10.1107/S1600536810035178/bg2359Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The central AgI ion is coordinated by two different N atoms in the aromatic rings of the ligands, with an N\u2014Ag\u2014N angle of 173.9\u2005(2)\u00b0. The crystal structure is composed of two complexed cations and stabilized by an inter\u00admolecular N\u2014H\u22efO and N\u2014H\u22efN hydrogen-bond network and there is \u03c0\u2013\u03c0 stacking of the aromatic rings [inter\u00adplanar distance 3.651\u2005(10)\u2005\u00c5].Colourless crystals of the title compound, [Ag(C DOI: 10.1107/S1600536809035181/jh2100Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, centrosymmetric dimers are formed through \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.624\u2005(2)\u2005\u00c5] between the substituted ferrocene ring and the three fused rings of the naphthalic anhydride unit. Pairs of dimers are held together by further naphthalene\u2013naphthalene \u03c0\u2013\u03c0 interactions [distance between parallel mean planes 3.45\u2005(3)\u2005\u00c5]. Each dimer inter\u00adacts with four neighbouring dimers in a herringbone fashion through C\u2014H\u22ef\u03c0 inter\u00adactions, so forming a two-dimensional sheet-like structure.In the structure of the title compound, [Fe(C DOI: 10.1107/S1600536808007393/su2047Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Mo\u2014Mo distance is 2.5732\u2005(4)\u2005\u00c5. The perpendicular distances of the Mo atoms from the C7 planes are 1.5827\u2005(8) and 1.5814\u2005(8)\u2005\u00c5, with individual Mo\u2014C bond lengths in the range 2.261\u2005(2)\u20132.2789\u2005(14)\u2005\u00c5. Mo\u2014H distances range from 1.77\u2005(3) to 1.85\u2005(4)\u2005\u00c5, with Mo\u2014H\u2014Mo angles of 89\u2005(2) and 92\u2005(1)\u00b0.In the title compound, [Mo DOI: 10.1107/S1600536808042244/bt2832Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CoII(TPP) complex exhibits a nearly planar conformation of the porphyrinate core [maximum deviation = 0.069\u2005(2)\u2005\u00c5] with an average Co\u2014N distance of 1.971\u2005(4)\u2005\u00c5. The distance between the Co atom and the closest O atom of the 18-crown-6 mol\u00adecule is 2.533\u2005(2)\u2005\u00c5, indicating a short non-bonded contact between the Co atom and the crown ether mol\u00adecule. An ethyl\u00adene group of the 18-crown-6 mol\u00adecule is disordered over two sites with occupancies of 0.565\u2005(7) and 0.435\u2005(7).The asymmetric unit of the title compound, [Co(C DOI: 10.1107/S1600536810012080/pv2268Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angle formed by the pyridine and benzene rings is 74.61\u2005(5)\u00b0. Intra\u00admolecular C\u2014H\u22efO hydrogen bonds are present. The crystal structure is stabilized by weak aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions involving neighbouring pyridine rings [centroid\u2013centroid distance = 3.853\u2005(2)\u2005\u00c5].In the mononuclear title complex, [Cu(C DOI: 10.1107/S1600536809029523/rz2354Isup2.hkl Structure factors: contains datablocks I. DOI:"} +{"text": "In the crystal structure, there is a weak \u03c0\u2013\u03c0 stacking inter\u00adaction between adjacent 1,10-phenanthroline rings, with a pyridine centroid\u2013centroid distance of 3.6620\u2005(15)\u2005\u00c5.In the title complex, [Zn(NCS) DOI: 10.1107/S1600536808011793/lh2603Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal packing is stabilized by \u03c0\u2013\u03c0 ; C\u2014H\u22ef\u03c0\u22eftolyl ring inter\u00adactions are also present.In the mol\u00adecule of the title compound, C DOI: 10.1107/S1600536808023222/dn2368Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized via inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine and benzene rings of neighboring phen ligands .In the title mononuclear complex, [Cu(C DOI: 10.1107/S1600536809053483/hy2263Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The N\u2014Ni\u2014N angle is 78.64\u2005(8)\u00b0. Weak but significant \u03c0\u2013\u03c0 stacking inter\u00adactions exist between the pyridine rings with a centroid\u2013centroid distance of 3.652\u2005(8)\u2005\u00c5. In addition, four O atoms of the two carboxyl groups form hydrogen bonds with both coordinated and uncoordinated water mol\u00adecules, forming an infinite three-dimensional network.In the title compound, [Ni(C DOI: 10.1107/S1600536809035910/ng2636Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The uncoordinated water mol\u00adecules are involved in O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, which contribute to the formation of a three-dimensional supra\u00admolecular structure, along with \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances of 3.5313\u2005(13) and 3.6028\u2005(11)\u2005\u00c5 between the pyridine rings of neighbouring dianions].In the title compound, (C DOI: 10.1107/S1600536808029395/cv2442Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Cu\u2014Cu distance is 2.6458\u2005(4)\u2005\u00c5. In the crystal structure, the dinuclear units are linked into a three-dimensional network by O\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds. One of the Cl atoms is disordered over two positions with occupancies of 0.650\u2005(2) and 0.350\u2005(2).In the title binuclear copper(II) complex, [Cu DOI: 10.1107/S1600536808041986/ci2732Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "An intra\u00admolecular C\u2014H\u22efO inter\u00adaction forms an S(7) ring motif. The dihedral angles between the benzoate group and the other three phenyl rings are 76.94\u2005(8), 66.82\u2005(8) and 42.34\u2005(9)\u00b0. The crystal structure is further stabilized by inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, [Sn(C DOI: 10.1107/S1600536808036337/kp2195Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by weak non-classical inter\u00admolecular C\u2014H\u22efO hydrogen-bond inter\u00adactions. The crystal structure also exhibits aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions between furan/benzene and benzene/benzene rings of adjacent benzofuran ring systems [centroid\u2013centroid distances = 3.8258\u2005(9) and 3.8794\u2005(9)\u2005\u00c5] and a weak inter\u00admolecular C\u2014H\u22ef\u03c0 ring inter\u00adaction.In the title compound, C DOI: 10.1107/S1600536809030451/jj2003Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure exhibits aromatic \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of adjacent mol\u00adecules [centroid\u2013centroid distance = 3.643\u2005(2)\u2005\u00c5] and nonclassical C\u2014H\u22efO hydrogen bonds.In the title compound, C DOI: 10.1107/S1600536809020613/cv2569Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "All six 4-methoxy\u00adbenzoate ligands act in a bidentate mode, two coordinating to one Tb center each and the other four bridging two Tb centers [Tb\u22efTb separation = 4.3144\u2005(6)\u2005\u00c5]. In the crystal, inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings of 1,10-phenanthroline and 4-methoxy\u00adbenzoate ligands [centroid\u2013centroid distance = 3.742\u2005(9)\u2005\u00c5] link two mol\u00adecules into a centrosymmetric dimer. Weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds help to consolidate the crystal packing.In the title dinuclear complex, [Tb DOI: 10.1107/S1600536809037751/cv2614Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The four O atoms in the equatorial plane around the CoII ion form a slightly distorted square-planar arrangement with an average Co\u2014O bond length of 2.086\u2005\u00c5; the slightly distorted octa\u00adhedral coordination is completed by the two N atoms of the isonicotinamide (INA) ligands at a slightly longer distance [2.1603\u2005(14)\u2005\u00c5] in the axial positions. The dihedral angle between the carboxyl\u00adate group and the attached benzene ring is 5.93\u2005(13)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 3.09\u2005(6)\u00b0. In the crystal structure, O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network. \u03c0\u2013\u03c0 Contacts between the benzene and pyridine rings [centroid\u2013centroid distance = 3.758\u2005(1)\u2005\u00c5] may further stabilize the crystal structure.The asymmetric unit of the crystal structure of the title complex, [Co(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(H2O)4](C8H5O3)2\u00b72H2O = 0.034 wR(F 2) = 0.074 S = 0.98 3780 reflections236 parameters9 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.36 e \u00c5\u22123 \u0394\u03c1min = \u22120.41 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008Mercury (Macrae et al., 2006WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S1600536809033200/xu2594sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536809033200/xu2594Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The anion is a six-coordinate complex with a distorted octa\u00adhedral geometry around the FeIII atom. A wide range of non-covalent inter\u00adactions, i.e. O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds, ion pairing, C\u2014O\u22ef\u03c0 [3.431\u2005(2)\u2005\u00c5] and C\u2014H\u22ef\u03c0 stacking inter\u00adactions result in the formation of a three-dimensional network structure.In the title compound, (C DOI: 10.1107/S1600536810008597/su2166Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each Cu atom is coordinated by a bidentate chelating 2,2\u2032-bipyridine ligand. A bridging cyanide anion links the two Cu units to form a binuclear unit. Adjacent binuclear units are connected by a thio\u00adcyanate anion into a one-dimensional helical chain along [010]. The cyanide anion is disordered, with each site occupied by both C and N atoms in an occupancy ratio of 0.61\u2005(5):0.39\u2005(5). The S atom of the thio\u00adcyanate anion is also disordered over two sites, with occupancy factors of 0.61\u2005(3) and 0.39\u2005(3). There are \u03c0\u2013\u03c0 inter\u00adactions between the pyridyl rings of neighbouring chains [centroid\u2013centroid distance = 3.82\u2005(1)\u2005\u00c5].The title compound, [Cu DOI: 10.1107/S1600536808037756/hy2162Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Numerous weak C\u2014H\u22efCl hydrogen bonds between [AuCl4]\u2212 and the 1,3-benzothia\u00adzolium units form layers comprised of 24-membered rings in which hydrogen-bonded tetra\u00adhydro\u00adfuran (THF) solvent mol\u00adecules are accommodated. C\u2014H\u22efCl inter\u00adactions between THF and [AuCl4]\u2212 from adjacent layers result in bilayers. These are further stabilized by \u03c0\u2013\u03c0 inter\u00adactions between the thia\u00adzole and benzene rings [centroid\u2013centroid distance = 3.971\u2005(3)\u2005\u00c5], resulting in the formation of a three-dimensional supra\u00admolecular assembly.In the crystal structure of the title ionic compound (C DOI: 10.1107/S1600536809003572/ng2541Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Cu and Pd atoms are both located on centers of symmetry in an alternating array of [Cu(N-Eten)2]2+ (N-Eten = N-ethyl\u00adethylenediamine) and [Pd(CN)4]2\u2212 units. The Pd\u2014C distances of 1.991\u2005(3) and 1.992\u2005(3)\u2005\u00c5 are inter\u00admediate values compared with the analogous NiII and PtII complexes [Akitsu & Einaga \u2005\u00c5 is noticeably longer than the equatorial distances [Cu\u2014NH2 = 2.007\u2005(2) and Cu\u2014NHC2H5 = 2.050\u2005(2)\u2005\u00c5]. There are interchain hybrogen bonds, with N(\u2014H)\u22efN = 3.099(4)\u2005\u00c5.The title compound, [CuPd(CN)naga 2007. Inorg. DOI: 10.1107/S160053680900885X/bg2245Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "This results in a distorted cis-ZnO2N4 octa\u00adhedral coordination geometry for the metal ion. In the crystal, mol\u00adecules are expanded into a three-dimensional supra\u00admolecular motif via O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22ef hydrogen bonds. In addition, \u03c0\u2013\u03c0 stacking inter\u00adactions between the aromatic rings of the polycyclic ligands consolidate the sturcture [shortest centroid\u2013centroid distance = 3.501\u2005(2)\u2005\u00c5].In the title compound, [Zn(C DOI: 10.1107/S1600536809039154/hb5120Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The three phosphine-substituted benzene rings make dihedral angles of 77.7\u2005(3), 84.4\u2005(3) and 77.4\u2005(3)\u00b0 with each other. Two of the three F atoms are disordered over two positions, with refined site occupancies of 0.591\u2005(11):0.409\u2005(11) and 0.730\u2005(12):0.270\u2005(12). In the crystal structure, mol\u00adecules are linked into a three-dimensional network by inter\u00admolecular C\u2014H\u22efCl and C\u2014H\u22efF hydrogen bonds.In the title gold complex, [AuCl(C DOI: 10.1107/S1600536810034896/fj2329Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ZnII atoms are four-coordinated in distorted tetra\u00adhedral configurations by two N atoms from 5,5\u2032-dimethyl-2,2\u2032-bipyridine and two terminal Cl atoms. In the crystal structure, inter\u00admolecular C\u2014H\u22efCl hydrogen bonds link the mol\u00adecules. There are C\u2014H\u22ef\u03c0 contacts between the methyl groups and the pyridine and five-membered rings containing ZnII atoms; \u03c0\u2013\u03c0 contacts also exist between the pyridine rings [centroid\u2013centroid distances = 3.665\u2005(5) and 3.674\u2005(5)\u2005\u00c5].The asymmetric unit of the title compound, [ZnCl DOI: 10.1107/S1600536808027104/hk2517Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the complex, the two cyclo\u00adpenta\u00addienyl (Cp) rings are almost parallel with a dihedral angle of 1.36\u2005(8)\u00b0, and are separated by a centroid\u2013centroid distance of 3.299\u2005(2)\u2005\u00c5. In the crystal, adjacent mol\u00adecules are linked into a one-dimensional supra\u00admolecular structure via weak C\u2014H\u22ef\u03c0 inter\u00adactions between the Cp ring H atom and the Cp ring.The title compound, [Fe(C DOI: 10.1107/S1600536810005155/hy2280Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The ZrIV centre is in a distorted tetra\u00adhedral environment defined by two Cp rings of chelating organic ligands and two Br anions. Two five-membered rings form a dihedral angle of 59.7\u2005(2)\u00b0. Unequal Zr\u2014C bonds [2.471\u2005(3)\u20132.556\u2005(3)\u2005\u00c5] in the mol\u00adecule indicate that the inter\u00adaction of the central metal with the [(C5H4)2SiMe2]2\u2212 ligand contains noticeable \u03b73-allyl and \u03b72-olefin contributions.The title mol\u00adecule, [ZrBr DOI: 10.1107/S1600536808040713/cv2494Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Ru\u2014Cl distances are 2.4161\u2005(7) and 2.4317\u2005(7)\u2005\u00c5, the Ru\u2014O distance is 2.1540\u2005(19)\u2005\u00c5, and the Ru\u2014S distances are in the range 2.2254\u2005(8)\u20132.2657\u2005(7)\u2005\u00c5, with the shortest being that trans to the aqua ligand. The coordinated water mol\u00adecule forms inter\u00admolecular hydrogen bonds with Cl and sulfoxide O atoms.The title mol\u00adecule, [RuCl DOI: 10.1107/S1600536809000439/pv2128Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The almost planar thio\u00adpyrano\u00adindole\u00adthione ring system makes a dihedral angle of 80.70\u2005(8)\u00b0 with the p-tolyl ring. In the crystal, mol\u00adecules are connected via C\u2014H\u22efO hydrogen bonds into two chains along the b axis. These chains are connected via \u03c0\u2013\u03c0 inter\u00adactions between symmetry-related thio\u00adpyrano\u00adindole\u00adthione ring systems [centroid\u2013centroid distance = 3.588\u2005(1)\u2005\u00c5].The title compound, C DOI: 10.1107/S1600536810038201/bt5362Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Mol\u00adecules are linked by C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and by \u03c0\u2013\u03c0 inter\u00adactions with inter\u00adplanar distances of 3.2661\u2005(6) and 3.2775\u2005(6)\u2005\u00c5.In the title compound, C DOI: 10.1107/S160053680803941X/fj2169Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each [CuBr4]2\u2212 anion is connected non-symmetrically to four surrounding cations through N\u2014H\u22efX (pyridine and amine proton) hydrogen bonds, forming chains of the ladder-type running parallel to the crystallographic b axis. These layers are further connected by means of offset face-to-face inter\u00adactions , giving a three-dimensional network. Cation \u03c0\u2013\u03c0 stacking [centroid separations of 3.69\u2005(9) and 3.71\u2005(1)\u2005\u00c5] and Br\u22efaryl inter\u00adactions [3.72\u2005(2) and 4.04\u2005(6)\u2005\u00c5] are present in the crystal structure. There are no inter\u00admolecular Br\u22efBr inter\u00adactions.In the crystal structure of the title compound, (C DOI: 10.1107/S1600536808010647/at2561Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, the cations and anions are linked by N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds, forming a layer network parallel to the ac plane. Weak \u03c0\u2013\u03c0 inter\u00adactions between adjacent pyridine rings with a centroid\u2013centroid distance of 3.589\u2005(2)\u2005\u00c5 are also present.In the dication of the title salt, C DOI: 10.1107/S1600536809038689/ng2649Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecule contains an inversion center in the middle of the Ge4O2 ring which is in a chair conformation. The Ge\u2014Ge bond distance is 2.4418\u2005(5)\u2005\u00c5 and the Ge\u2014O bond distances are 1.790\u2005(2) and 1.785\u2005(2)\u2005\u00c5. The torsion angles within the Ge4O2 ring are \u221256.7\u2005(1) and 56.1\u2005(1)\u00b0 for the Ge\u2014Ge\u2014O\u2014Ge angles and \u221243.9\u2005(1)\u00b0 for the O\u2014Ge\u2014Ge\u2014O angle.The title compound, C DOI: 10.1107/S1600536809032012/lh2875Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The compound displays intra\u00admolecular C\u2014H\u22efCl hydrogen bonds and pairs of complex mol\u00adecules are connected by inter\u00admolecular C\u2014H\u22efCl hydrogen bonds. Inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions are present between the pyridine rings of the ligand, the shortest centroid\u2013centroid distance being 4.096\u2005(3)\u2005\u00c5. As a result of the electronic nature of the chelate ring, it is possible to create \u03c0\u2013\u03c0 inter\u00adactions to its symmetry-related counterpart [3.720\u2005(2)\u2005\u00c5] and also with a pyridine ring [3.570\u2005(3)\u2005\u00c5] of the bipy unit. The present structure is a redetermination of a previous structure [Vicente et al. (1997In the title compound, [PdCl al. 1997. Private X 2(bipy)] (X = Cl or Br), see: Maekawa et al. ]\u00b7CH2Cl2 = 0.044 wR(F 2) = 0.097 S = 1.05 2862 reflections203 parametersAll H-atom parameters refinedmax = 0.56 e \u00c5\u22123 \u0394\u03c1min = \u22120.69 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 (Farrugia, 1997PLATON (Spek, 2009SHELXL97.Data collection: 10.1107/S1600536809016262/kp2220sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536809016262/kp2220Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, 2-amino-3-(1H-indol-3-yl)propionic acid mol\u00adecules and pyridine-2,4-dicarb\u00adoxylic acid mol\u00adecules are linked through strong inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming layers parallel to (100). The layers are linked through the ethanol mol\u00adecules via somewhat weaker inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming thus a three-dimensional network. Weak C\u2014H\u22efO and N\u2014H\u22efN hydrogen bonding and \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings are also present.In the title compound, C DOI: 10.1107/S1600536810014017/fb2190Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between the 1,10-phenanthroline ligands and 5-chloro\u00adsalicylate ligands, with a centroid\u2013centroid distance between neighbouring aromatic rings of 3.730\u2005(1)\u2005\u00c5.In the title complex, [Cd(C DOI: 10.1107/S1600536808015687/wn2261Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The seven donor atoms of the (pydc)2\u2212 groups and the hydroxido ligands form a distorted penta\u00adgonal\u2013bipyramidal arrangement around the SbIII/SbV centers. C\u2014H\u22ef\u03c0 stacking inter\u00adactions between CH groups of the complex dianion and the aromatic rings of the + cations, with a distance of 2.89\u2005\u00c5, are observed. In the crystal structure, a wide range of noncovalent inter\u00adactions, consisting of O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds [D\u22efA ranging from 2.722\u2005(2) to 3.137\u2005(3)\u2005\u00c5], ion pairing, \u03c0\u2013\u03c0 stacking [centroid\u2013centroid distance of 3.4363\u2005(13)\u2005\u00c5] and C\u2014H\u22ef\u03c0 inter\u00adactions, connect the various components into a supra\u00admolecular structure.The reaction of anti\u00admony(III) chloride, 4,4\u2032-bipyridine and pyridine-2,6-dicarboxylic acid (pydcH DOI: 10.1107/S1600536808001372/su2034Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "All three bidentate ligands have coordinating ring systems that are almost coplanar [inter\u00adplanar angles = 1.7\u2005(1)\u20133.8\u2005(2)\u00b0]. The vinyl\u00adbenzyl group is disordered over two positions with occupations of 0.653\u2005(4) and 0.347\u2005(4). The methanol solvent mol\u00adecule is involved in a classical O\u2014H\u22efN hydrogen bond to a triazole N atom.In the title compound, [Ir(C DOI: 10.1107/S1600536809052726/tk2595Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The torsion angle for the Te\u2014Re\u2014Te\u2014Re sequence of atoms is 17.06\u2005(3)\u00b0. The crystal structure is stabilized by weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. In addition, there are Te\u22efTe distances [4.0392\u2005(12)\u2005\u00c5] and O\u22efO distances [2.902\u2005(19)\u2005\u00c5] which are shorter than the sum of the van der Waals radii for these atoms. A short inter\u00admolecular lone pair\u22ef\u03c0 distance [C\u00a0O\u22efCg = 3.31\u2005(2)\u2005\u00c5] is also observed.In the title complex, [Re DOI: 10.1107/S1600536810012389/lh5009Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by aromatic \u03c0\u2013\u03c0 inter\u00adactions, with centroid\u2013centroid distances of 3.765\u2005(5) and 3.896\u2005(5)\u2005\u00c5 between the phenyl ring and the benzene ring of the chromene unit of neighbouring mol\u00adecules. In addition, the stacked mol\u00adecules exhibit inter- and intra\u00admolecular O\u2014H\u22efO hydrogen bonds, including the uncoordinated water mol\u00adecules.In the title compound, [Ca(C DOI: 10.1107/S1600536808037586/lx2072Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two EuIII ions are linked by four bridging benzoate ions, with an Eu\u22efEu distance of 3.96041\u2005(12)\u2005\u00c5. Each EuIII ion is coordinated by one phenanthroline heterocycle and a bidentate benzoate ion. The irregular nine-coordinated geometry of the metal ion is composed of seven O and two N atoms. The mol\u00adecular structure is stabilized by intra\u00admolecular C\u2014H\u22efO hydrogen bonds. In the crystal structure, mol\u00adecules are linked into chains by inter\u00admolecular C\u2014H\u22efO hydrogen bonds along the a axis. The crystal structure is further stabilized by inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. Weak \u03c0\u2013\u03c0 inter\u00adactions are also observed [centroid\u2013centroid distances = 3.6962\u2005(10)\u20133.6963\u2005(10)\u2005\u00c5].The asymmetric unit of the title complex, [Eu For rel al. 1999, 2002 \u25b6;2(C7H5O2)6(C12H8N2)2]\u00b72C7H6O2 = 0.020 wR(F 2) = 0.050 S = 1.04 12557 reflections464 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.62 e \u00c5\u22123 \u0394\u03c1min = \u22120.44 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXTL (Sheldrick, 2008SHELXTL; molecular graphics: SHELXTL; software used to prepare material for publication: SHELXTL and PLATON (Spek, 2009Data collection: 10.1107/S1600536810015229/rz2435sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536810015229/rz2435Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "All intramolecular distances and angles are similar for the two structures. This applies also for the intermolecular forces, consisting of O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 interactions [with centroid\u2013centroid distances of 3.428\u2005(2) and 3.579\u2005(2)\u2005\u00c5], that lead to a cohesion of the structure.The title compound, (C DOI: 10.1107/S1600536809022053/pv2161Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The main geometric parameters are Pt\u2014N = 1.960\u2005(5)\u2005\u00c5, and Pt\u2014Cl = 2.3177\u2005(12) and 2.3196\u2005(12)\u2005\u00c5. The N\u00a0C bond is a typical triple bond [1.137\u2005(7)\u2005\u00c5]. The Pt\u2014N\u00a0C\u2014C unit is almost linear, with Pt\u2014N\u2014C and N\u2014C\u2014C angles of 174.6\u2005(4) and 177.1\u2005(6)\u00b0, respectively.In the title compound, [PtCl DOI: 10.1107/S1600536809016535/pv2142Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, inter\u00admolecular bifurcated N\u2014H\u22ef hydrogen bonds connect the mol\u00adecules into chains propagating along [100]. Adjacent chains inter\u00adact by strong aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions, with a centroid\u2013centroid distance of 3.483\u2005(2)\u2005\u00c5.In the title compound, [Pb(NO DOI: 10.1107/S1600536808041317/hb2871Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The conformations of the two crystallographically independent mol\u00adecules are slightly different: in A, the Cu\u22efCu separation is 4.174\u2005(9)\u2005\u00c5 and the dihedral angle between the triazole and phenyl rings is 74.23\u2005(11)\u00b0; these values are 4.137\u2005(9)\u2005\u00c5 and 68.58\u2005(13)\u00b0, respectively, in B. In each mol\u00adecule, the copper(II) ions have a distorted trigonal\u2013bipyramidal coordination geometry with a CuCl2NN\u2032N\u2032\u2032 chromophore. The crystal packing exhibits weak inter\u00admolecular C\u2014H\u22efCl inter\u00adactions.The asymmetric unit of the title compound, [Cu DOI: 10.1107/S160053680804035X/cv2472Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The K\u22efO distances range from 2.8416\u2005(8) to 3.0025\u2005(8)\u2005\u00c5, and the two K\u22efN distances are 2.7441\u2005(11) and 2.7654\u2005(11)\u2005\u00c5. The K cation is displaced by 0.8437\u2005(4)\u2005\u00c5 from the best plane through the six O atoms. The latter plane is almost perpendicular to the plane of the pyrazolate ring [dihedral angle 83.93\u2005(3)\u00b0].The asymmetric unit of the title compound, [K(C DOI: 10.1107/S1600536808043237/rz2281Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The other Sn atom has a bent R 2Sn skeleton [C\u2014Sn\u2014C = 144.0\u2005(1)\u00b0], but the geometry is best regarded as being a trans-C2SnO4 octa\u00adhedron as the Sn\u2013O(single bond) inter\u00adaction is shorter [Sn\u2014O = 2.674\u2005(1)\u2005\u00c5].In the centrosymmetric tetra\u00adnuclear title compound, [Sn DOI: 10.1107/S1600536808023787/tk2287Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Both the phosphine and arsine ligands are equatorial with respect to the Ru3 triangle. In addition, each Ru atom carries one equatorial and two axial terminal carbonyl ligands. All three cyclo\u00adhexane rings are disordered over two positions with site occupancies of 0.628\u2005(6) and 0.372\u2005(6). The mean planes of these three phosphine-substituted cyclo\u00adhexane rings make dihedral angles of 53.0\u2005(8), 68.3\u2005(6) and 89.9\u2005(7)\u00b0 (major components), and 46.7\u2005(14), 41.3\u2005(11) and 75.8\u2005(10)\u00b0 (minor components) with each other. The dihedral angles between the two phenyl rings are 85.0\u2005(2) and 88.1\u2005(2)\u00b0 for the two diphenyl\u00adarsino groups. Two cyclo\u00adhexane rings adopt a chair conformation whereas the other adopts a slightly twisted chair conformation for the major components; these conformations are similiar for the minor components. Intra\u00admolecular C\u2014H\u22efO hydrogen bonds stabilize the mol\u00adecular structure. In the crystal packing, the mol\u00adecules are linked together into chains via inter\u00admolecular C\u2014H\u22efO hydrogen bonds down the a axis. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions further stabilize the crystal structure.In the title DOI: 10.1107/S1600536809047977/sj2677Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The crystal structure is stabilized by a combination of weak \u03c0\u2013\u03c0 stacking inter\u00adactions between symmetry-related 1,10-phenanthroline ligands [centroi\u2013centroid distance between benzene rings = 3.5936\u2005(18)\u2005\u00c5] and weak O\u2014H\u22efS, C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds between methanol and complex mol\u00adecules.In the title complex, [Ni(NCS) DOI: 10.1107/S1600536808011811/lh2613Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each DyIII ion is coordinated by six O atoms and two N atoms from two pyridine-2,6-dicarboxyl\u00adate and two 6-carboxy\u00adpicolinate ligands, displaying a bicapped trigonal-prismatic geometry. The average Dy\u2014O bond distance is 2.40\u2005\u00c5, some 0.1\u00c5 longer than the corresponding Ho\u2014O distance in the isotypic holmium complex. Adjacent DyIII ions are linked by the pyridine-2,6-dicarboxyl\u00adate ligands, forming a layer in (100). These layers are further connected by \u03c0\u2013\u03c0 stacking inter\u00adactions between neighboring pyridyl rings [centroid\u2013centroid distance = 3.827\u2005(3)\u2005\u00c5] and C\u2014H\u22efO hydrogen-bonding inter\u00adactions, assembling a three-dimensional supra\u00admolecular network. Within each layer, there are other \u03c0\u2013\u03c0 stacking inter\u00adactions between neighboring pyridyl rings [centroid\u2013centroid distance = 3.501\u2005(2)\u2005\u00c5] and O\u2014H\u22efO and C\u2014H\u22efO hydrogen-bonding inter\u00adactions, which further stabilize the structure.In the title complex, [Dy(C DOI: 10.1107/S1600536809039075/sj2654Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, mol\u00adecules are stacked alternately in opposite orientations, forming a one-dimensional column parallel to [110]. The primary inter\u00adactions between mol\u00adecules comprising the columns are of the S\u22efS type [3.554\u2005(1)\u2005\u00c5]. Inter\u00adactions between columns are of the S\u22efS type [3.411\u2005(1) along b and 3.444\u2005(1)\u2005\u00c5 along c], as well as S\u22efI contacts [3.435\u2005(2)\u2005\u00c5].The mol\u00adecular framework of the title compound, C DOI: 10.1107/S1600536809044493/tk2559Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "These units are connected via the carboxyl\u00adate O atoms and water mol\u00adecules, building polymeric layers parallel to (100). In the crystal structure, non-covalent inter\u00adactions consisting of O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.862\u2005(17) and 3.749\u2005(17)\u2005\u00c5] connect the various components, forming a three-dimensional structure.In the title polymeric complex, [Sr(C DOI: 10.1107/S160053680902683X/pv2167Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each CuI atom is tetra\u00adhedrally coordinated by three I atoms and one N atom of an benzyl\u00addimethyl\u00adamine ligand. Each I atom acts as a \u03bc3-ligand, linking three CuI atoms. The Cu\u2014I bond distances vary between 2.6328\u2005(7) and 2.7121\u2005(6)\u2005\u00c5, while the Cu\u2014N bond distances vary between 2.107\u2005(3) and 2.122\u2005(3)\u2005\u00c5.The title compound, [Cu DOI: 10.1107/S1600536809026208/fj2223Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII atom exhibits a six-coordinated distorted octa\u00adhedral geometry with two N atoms from the phenanthroline ligand [Cu\u2014N 2.007\u2005(2)\u2005\u00c5] and four O atoms from two 3,4-dimethoxy\u00adbenzoate ligands [Cu\u2014O 1.950\u2005(1) and 2.524\u2005(1)\u2005\u00c5]. The difference in Cu\u2014O bond distances indicates a strong Jahn\u2013Teller effect. In the crystal, C\u2014H\u22ef\u03c0 inter\u00adactions result in chains of mol\u00adecules along the c axis.The asymmetric unit of the title compound, [Cu(C DOI: 10.1107/S1600536809052234/ez2194Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The phenyl rings are rotated out of the benzofuran planes, making dihedral angles of 43.38\u2005(7) and 56.13\u2005(6)\u00b0 in the two mol\u00adecules. The carboxyl groups are involved in inversion-related inter\u00admolecular O\u2014H\u22efO hydrogen bonds, which link the mol\u00adecules into centrosymmetric dimers. These dimers are further packed into stacks along the b axis by weak non-classical inter\u00admolecular C\u2014H\u22efO hydrogen bonds. The crystal structure also exhibits inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions, and two aromatic \u03c0\u2013\u03c0 inter\u00adactions between the furan rings of neighbouring benzofuran systems; the centroid\u2013centroid distances are 3.500\u2005(3) and 3.605\u2005(3)\u2005\u00c5.The title compound, C DOI: 10.1107/S160053680903253X/nk2002Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Cu atom is five-coordinated in a distorted trigonal-bipyramidal geometry by two phen mol\u00adecules and one bromide ligand, which coordinates in the equatorial plane at a distance of 2.5228\u2005(4)\u2005\u00c5 and lying along with the Cu and the amide N atoms on a twofold rotation axis. The two axial Cu\u2014N distances [1.9926\u2005(15)\u2005\u00c5] are slightly shorter than the two equatorial Cu\u2014N bonds [2.0979\u2005(15)\u2005\u00c5]. The structure is stabilized by a weak C\u2014H\u22efN hydrogen bond, with a cyanide N atom of the dicyanamide ligand as an acceptor, and \u03c0\u2013\u03c0 inter\u00adactions between nearly parallel phenyl and pyridine rings of two adjacent phen mol\u00adecules [centroid\u2013centroid distance = 3.589\u2005(1)\u2005\u00c5], and between \u03c0 electrons of the dicyanamide anion and the pyridine ring .The title compound, [CuBr(C For pen al. 1984. For a d al. 1984. For ref al. 1984. 12H8N2)2]C2N3 = 0.021 wR(F 2) = 0.062 S = 1.06 2182 reflections160 parametersH-atom parameters constrainedmax = 0.29 e \u00c5\u22123 \u0394\u03c1min = \u22120.43 e \u00c5\u22123 \u0394\u03c1 CrysAlis CCD used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008DIAMOND (Brandenburg, 2001SHELXL97.Data collection: 10.1107/S1600536810037979/kp2274sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810037979/kp2274Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The C\u2014P bond lengths are 1.8824\u2005(19) and 1.8991\u2005(19)\u2005\u00c5. The P\u2014C\u2014P angle of 109.69\u2005(9)\u00b0 is essentially equal to the expected value of 109.5\u00b0 for a tetra\u00adhedral C atom. Although the C(methine)\u2014P\u2014C(aromatic) bond angles range from 102.67\u2005(9) to 107.04\u2005(9)\u00b0, the C(aromatic)\u2014P\u2014C(aromatic) bond angles of 96.72\u2005(9) and 97.29\u2005(9)\u00b0 are significantly smaller. The steric demands of the o-tolyl groups cause deviations from the bond lengths and angles reported for its phenyl analog.The complete molecule of title compound, C DOI: 10.1107/S1600536809032942/ng2629Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "A water mol\u00adecule cocrystallizes with the coordination compound and may be held in the crystal by O\u2014H\u22efO hydrogen bonds. Heteroatomic \u03c0\u2013\u03c0 ring inter\u00adactions may be present between symmetry-related complexes, with centroid\u2013centroid distances of 3.5661\u2005(8)\u2005\u00c5.In the title complex, [Co(C DOI: 10.1107/S1600536809021540/cs2106Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Adjacent stacked mol\u00adecules exhibit a carbon\u00adyl\u2013carbonyl inter\u00adaction [3.295\u2005(2)\u2005\u00c5]. The O atom of the methyl\u00adsulfinyl group is disordered over two positions with site-occupancy factors of 0.9 and 0.1.Mol\u00adecules of title compound, C DOI: 10.1107/S1600536808034466/ng2503Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Each CrIII atom is hexa\u00adcoordinated by four O and two N atoms from two (hypydc)2\u2212 fragments, which act as tridentate ligands, in a distorted octa\u00adhedral geometry. The O\u2014Cr\u2014O\u2014C torsion angles between the two planes of the (hypydc)2\u2212 fragments [\u221299.81\u2005(17) and 97.77\u2005(17)\u00b0] indicate that these two units are almost perpendicular to one another. In the crystal structure, extensive O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds with D\u22efA distances ranging from 2.560\u2005(2) to 3.279\u2005(3)\u2005\u00c5, ion pairing, C\u2014O\u22ef\u03c0 [O\u22ef\u03c0 = 3.166\u2005(2)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking inter\u00adactions between (hypydc)2\u2212 and (pydaH)+ rings [with a centroid\u2013centroid distance of 3.3353\u2005(14)\u2005\u00c5] contribute to the formation of a three-dimensional supra\u00admolecular structure.The reaction of chromium(III) nitrate hexa\u00adhydrate, pyridine-2,6-diamine and 4-hydroxy\u00adpyridine-2,6-dicarboxylic acid in a 1:2:2 molar ratio in aqueous solution resulted in the formation of the title compound, (C DOI: 10.1107/S1600536808027347/ym2072Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit contains one mol\u00adecule of the complex and half a water solvent mol\u00adecule. The solvent water mol\u00adecule lies on a crystallographic twofold rotation axis. An inter\u00admolecular O\u2014H\u22efO hydrogen bond forms an R 2 1(4) ring motif involving a bifurcated hydrogen bond to the phenolate O atoms of the complex. In the crystal structure, mol\u00adecules are linked by \u03c0\u2013\u03c0 stacking inter\u00adactions, with centroid\u2013centroid distances in the range 3.5310\u2005(11)\u20133.7905\u2005(12)\u2005\u00c5, forming extended chains along the b axis. In addition, there are Ni\u22efNi and Ni\u22efN inter\u00adactions which are shorter than the sum of the van der Waals radii of the relevant atoms. Further stabilization of the crystal structure is attained by weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions.In the title complex, [Ni(C DOI: 10.1107/S1600536808023362/lh2663Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit of the title compound comprises one-half of the complex mol\u00adecule and one of the water mol\u00adecules of crystallization. The water H atoms form bifurcated O\u2014H\u22ef hydrogen bonds with the O atoms of the phenolato and eth\u00adoxy groups with R 1 2(5) and R 1 2(6) ring motifs. The dihedral angle between the central benzene ring and the two outer benzene rings are 4.07\u2005(11) and 3.99\u2005(12)\u00b0. The dihedral angle between the two O\u2013Ni\u2013N coordination planes is only 0.77\u2005(11)\u00b0. In the crystal structure, the mol\u00adecules are linked together into extended chains along the c axis by inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions. An inter\u00adesting feature of the crystal structure is a short inter\u00admolecular C \u22ef C [3.355\u2005(3)\u2005\u00c5] contact, which is shorter than the sum of the van der Waals radii. The crystal structure may be further stabilized by inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances in the range 3.5758\u2005(13)\u20133.6337\u2005(13)\u2005\u00c5]. In the title complex, [Ni(C DOI: 10.1107/S1600536809012641/cs2114Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, centrosymmetric dimers are formed though O\u2014H\u22efO and O\u2014H\u22efCl hydrogen bonds; \u03c0\u2013\u03c0 stacking inter\u00adactions, with a centroid\u2013centroid distance of 3.796\u2005(2)\u2005\u00c5, are also found.In the title compound, [Ni(C DOI: 10.1107/S1600536808022939/lh2644Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the cation, the Cu2+ ion is coordinated by four N atoms from two bidentate 2,2\u2032-bipyridine mol\u00adecules and an N atom from an isothio\u00adcyanate anion, resulting in a distorted CuN5 pyramidal configuration. The crystal structure is stabilized by weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds, and weak \u03c0\u2013\u03c0 inter\u00adactions between 2,2\u2032-bipyridine rings [centroid\u2013centroid distance = 3.908\u2005(4)\u2005\u00c5]. The perchlorate counteranion is disordered over two positions in a 0.66:0.34 ratio.The asymmetric unit of title compound, [Cu(NCS)(C DOI: 10.1107/S1600536809031067/at2856Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Ca2+ ions are bridged by 2-chloronicotinate groups and exhibit an eight-coordination by six carboxylate O atoms of four different 2-chloronicotinate ligands and two O atoms of water molecules. In the crystal structure, inter\u00admolecular O\u2014H\u22efO, O\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds result in the formation of a supra\u00admolecular network structure. The \u03c0\u2013\u03c0 contacts between the 2-chloro\u00adnicotinate rings [centroid\u2013centroid distances = 3.875\u2005(3) and 3.701\u2005(3)\u2005\u00c5] may further stabilize the structure. The itle compound, [Ca(C DOI: 10.1107/S1600536808044267/hk2578Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The L ligands chelate the LaIII ion with strong La\u2014O(deprotonated phenolic) bonds [2.435\u2005(3)\u20132.464\u2005(3)\u2005\u00c5] and significantly longer La\u2014O(meth\u00adoxy) bonds [2.801\u2005(3)\u20132.831\u2005(3)\u2005\u00c5]. The La\u2014N bond lengths range from 2.541\u2005(4) to 2.605\u2005(4)\u2005\u00c5.In the title compound, [La(NCS) DOI: 10.1107/S1600536809049113/pv2228Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The two rings are almost parallel and are eclipsed and ordered. The conjugated substituent is slightly twisted with respect to the cyclo\u00adpenta\u00addiene ring. The crystal structure contains four inter\u00admolecular C\u2014H\u22efO hydrogen-bonds in the range 3.324\u2005(3)\u20133.539\u2005(3)\u2005\u00c5 and one \u03c0(aryl ring)\u2013\u03c0 (Cp ring) stacking inter\u00adaction with a ring\u2013centroid distance of 3.894\u2005(2)\u2005\u00c5.In the title compound, [Fe(C DOI: 10.1107/S1600536808027815/om2257Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The plane containing the thio\u00adcarbonyl and carbonyl groups subtends dihedral angles of 48.19\u2005(3) and 87.51\u2005(3)\u00b0 with the planes formed by the 3-chloro and 2,6-dichloro\u00adphenyl rings, respectively; the dihedral angle between the two benzene ring planes is 45.32\u2005(3)\u00b0. An intra\u00admolecular N\u2014H\u22efO hydrogen bond stabilizes the mol\u00adecular conformation and the mol\u00adecules form inter\u00admolecular N\u2014H\u22efS and N\u2014H\u22efO hydrogen bonds, generating a sheet along the a axis.The structure of the title compound, C DOI: 10.1107/S1600536808043444/pv2127Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "One of the Cp rings is disordered over two positions with occupancies of 0.55 and 0.45. The dihedral angle between the substituted Cp rings is 56.1\u2005(5)\u00b0 and the two phenyl rings are orientated at a dihedral angle of 41.7\u2005(4)\u00b0. In the crystal structure, inter\u00admolecular O\u2014H\u22efS, N\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds link the mol\u00adecules into chains along the b axis. The structure is further consolidated by O\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, [Fe DOI: 10.1107/S1600536809046078/ci2959Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Intra\u00admolecular N\u2014H\u22efO hydrogen bonds occur. The crystal packing exhibits weak inter\u00admolecular C\u2014H\u22efS hydrogen bonds, \u03c0\u2013\u03c0 inter\u00adactions with a distance of 3.904\u2005(7)\u2005\u00c5 between the centroids of the aromatic rings, and voids of 101\u2005\u00c53.In the title compound, [Nd(NCS) DOI: 10.1107/S1600536809053124/cv2653Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The trinuclear mol\u00adecules, with an intramolecular Cu\u22efCu separation of 6.313\u2005(3)\u2005\u00c5, are linked to each other, forming a chain through O\u2014H\u22efO and O\u2014H\u22efBr hydrogen bonds involving the half-occupied water mol\u00adecule. Futhermore, weak C\u2014H\u22efO inter\u00adactions link the chains to form a supra\u00admolecular network.The title complex, [Cu DOI: 10.1107/S1600536809014263/dn2446Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The phenyl ring makes a dihedral angle of 83.64\u2005(4)\u00b0 with the mean plane of the 5,6-(methyl\u00adene\u00addi\u00adoxy)\u00adbenzo\u00adfuran fragment. The crystal structure is stabilized by C\u2014H\u22ef\u03c0 inter\u00adactions between a benzene H atom of the 5,6-(methyl\u00adene\u00addi\u00adoxy)\u00adbenzo\u00adfuran unit and the phenyl ring of the phenyl\u00adsulfonyl substituent. Additionally, the crystal structure exhibits inter- and intra\u00admolecular C\u2014H\u22efO inter\u00adactions.The title compound, C DOI: 10.1107/S160053680800980X/bh2167Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There are two uncoord\u00adinated water mol\u00adecules in the structure. The asymmetric unit of the compound has two different coordination types of ZnII in the crystal structure that are bridged to each other via one hypydc2\u2212 group . A variety of inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds involving water mol\u00adecules, cations and anions, and also a weak \u03c0\u2013\u03c0 inter\u00adaction [3.798\u2005(1)\u2005\u00c5], are responsible for extending the structure into a three-dimensional network.The title compound, (CH DOI: 10.1107/S1600536808003930/om2211Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The four nearest O atoms around each CuII ion form a distorted square-planar arrangement, and the distorted square-pyramidal coordination is completed by the pyridine N atom of the isonicotinamide (INA) ligand. Each CuII ion is displaced by 0.2633\u2005(1)\u2005\u00c5 from the plane of the four O atoms, with an average Cu\u2014O distance of 1.974\u2005(2)\u2005\u00c5. The dihedral angles between carboxyl\u00adate groups and the adjacent benzene rings are 7.88\u2005(19) and 9.68\u2005(10)\u00b0, while the benzene rings are oriented at a dihedral angle of 85.90\u2005(9)\u00b0. The pyridine ring is oriented at dihedral angles of 8.59\u2005(7) and 83.89\u2005(9)\u00b0 with respect to the benzene rings. In the crystal structure, inter\u00admolecular N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network. \u03c0\u2013\u03c0 contacts between the benzene rings and between the pyridine and benzene rings, may further stabilize the crystal structure.In the title centrosymmetric binuclear complex, [Cu N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 4(C6H6N2O)2] = 0.043 wR(F 2) = 0.087 S = 1.01 5101 reflections281 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.47 e \u00c5\u22123 \u0394\u03c1min = \u22120.70 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S1600536810006513/xu2729sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810006513/xu2729Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecule is centrosymmetric; the pairs of equivalent ligands coordinate trans to each other in a distorted octa\u00adhedral geometry. Intra\u00admolecular C\u2014H\u22efO bonds lying in the equatorial plane stabilize the mol\u00adecule. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, creating a three-dimensional supra\u00admolecular structure. \u03c0\u2013\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions are also observed. The dihedral angle and centroid-to-centroid distance between the pyridine ring (A) and the benzene ring (B i) of a symmetrically related mol\u00adecule are 1.27\u2005(11)\u00b0 and 3.974\u2005(2)\u2005\u00c5, respectively. For the C\u2014H\u22ef\u03c0 inter\u00adactions, the relevant distances and angles are: C\u22efCg[A ii] = 3.643\u2005(2)\u2005\u00c5, H\u22efCg[A ii] = 2.750\u2005(2)\u2005\u00c5 and C\u2014H\u22efCg[A ii] = 155\u2005(1)\u00b0 .The title compound, [Mn(C II complexes with the quinoline-2 carboxyl\u00adate ligand, see: Okabe &Koizumi 2(CH4O)2] = 0.034 wR(F 2) = 0.089 S = 0.98 1924 reflections146 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.37 e \u00c5\u22123 \u0394\u03c1min = \u22120.31 e \u00c5\u22123 \u0394\u03c1 CrysAlis CCD used to solve structure: SHELXTL-NT (Sheldrick, 2008SHELXTL-NT; molecular graphics: SHELXTL-NT; software used to prepare material for publication: SHELXTL-NT.Data collection: 10.1107/S1600536808031905/su2065sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536808031905/su2065Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The environment around the Ho atom can be described as a distorted bicapped square anti\u00adprism. Two phenanthroline ligands form a dihedral angle of 43.72\u2005(13)\u00b0. Short inter\u00admolecular distances between the centroids of the six-membered rings [3.6887\u2005(14)\u20133.8374\u2005(16)\u2005\u00c5] indicate the existence of \u03c0\u2013\u03c0 inter\u00adactions, which link the mol\u00adecules into stacks extended in the [10In the title compound, [Ho(NO DOI: 10.1107/S1600536808020655/cv2423Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Zn(C9H10NO2)2(C6H6N2O)(H2O)2], contains two 4-(dimethyl\u00adamino)benzoate (DMAB) ligands, one isonicotinamide (INA) ligand and two water mol\u00adecules; one of the DMAB ions acts as a bidentate ligand while the other and INA are monodentate ligands. The four O atoms in the equatorial plane around the Zn atom form a distorted square-planar arrangement, while the distorted octa\u00adhedral coordination is completed by the N atom of the INA ligand and the O atom of the water mol\u00adecule in the axial positions. Intra\u00admolecular C\u2014H\u22efO hydrogen bonding results in the formation of a six-membered ring adopting an envelope conformation. The dihedral angle between the carboxyl groups and the adjacent benzene rings are 4.87\u2005(16) and 2.2\u2005(2)\u00b0, while the two benzene rings are oriented at a dihedral angle of 65.13\u2005(8)\u00b0. The dihedral angle between the benzene and pyridine rings are 11.47\u2005(7) and 74.83\u2005(8)\u00b0, respectively. In the crystal structure, inter\u00admolecular O\u2014H\u22efO, O\u2014H\u22efN and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a supra\u00admolecular structure. \u03c0\u2013\u03c0 contacts between the pyridine and benzene rings and between the benzene rings further stabilize the structure. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions are also present.The mol\u00adecule of the title Zn DOI: 10.1107/S1600536809017620/xu2522Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordinated water mol\u00adecule is involved in inter\u00admolecular O\u2014H\u22efO hydrogen bonding, which links the complex mol\u00adecules into chains propagating along the c axis. Neighbouring chains inter\u00adact further via \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings of 1,10-phenanthroline fragments [centroid\u2013centroid distances = 3.726\u2005(4) and 3.750\u2005(4)\u2005\u00c5].The asymmetric unit of the title compound, [Cu(C DOI: 10.1107/S1600536809024428/cv2568Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The 4-tolyl ring makes a dihedral angle of 70.96\u2005(6)\u00b0 with the plane of the naphthofuran fragment. The crystal structure is stabilized by aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions, with centroid\u2013centroid distances of 3.672\u2005(3) and 3.858\u2005(3)\u2005\u00c5 between the central benzene and furan rings, and between the brominated benzene and central benzene rings of the naphthofuran system of neighbouring mol\u00adecules, respectively. In addition, the stacked mol\u00adecules exhibit C\u2014H\u22ef\u03c0 and inter- and intra\u00admolecular C\u2014H\u22efO inter\u00adactions.The title compound, C DOI: 10.1107/S1600536808015286/at2569Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by inter\u00admolecular N\u2014H\u22efO and N\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions of the aromatic rings of the two ligands [inter\u00adplanar distance = 3.652\u2005(10)\u2005\u00c5]. The AgI atom shows a linear coordination [N\u2014Ag\u2014N = 174.6\u2005(1)\u00b0].Colourless crystals of the title mixed ligand complex, [Ag(C DOI: 10.1107/S1600536809035193/ng2621Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Co(C5H3N2O4)2(H2O)4]\u00b72H2O, the CoII ion is located on an inversion center and is coordinated by two orotate anions and four water mol\u00adecules in a slightly distorted octa\u00adhedral geometry. The dihedral angle between the carboxyl\u00adate group and the attached orotate ring is 1.2\u2005(3)\u00b0. In the crystal structure, inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network. \u03c0\u2013\u03c0 contacts between the orotate rings [centroid\u2013centroid distances = 3.439\u2005(2) and 3.438\u2005(2)\u2005\u00c5] further stabilize the structure.In the title Co DOI: 10.1107/S1600536810015837/xu2754Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There is a \u03c0\u2013\u03c0 contact between pyridine rings of adjacent molecules [centroid\u2013centroid distance = 3.723\u2005(5)\u2005\u00c5].In the mol\u00adecule of the title compound, [HgI DOI: 10.1107/S160053680802953X/hk2531Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordination geometry for Cd is distorted octa\u00adhedral based on a cis-N2S4 donor set. In the crystal structure, mol\u00adecules are connected into a supra\u00admolecular chain aligned along the a direction via C\u2014H\u22efS and C\u2014H\u22ef\u03c0 contacts, and by \u03c0\u2013\u03c0 contacts [centroid-to-centroid distance 3.5708\u2005(15)\u2005\u00c5]. The n-propyl groups are each disordered, one equally over two sites and the other with a site-occupancy factor of 0.618\u2005(8) for the major component. The dinuclear centrosymmetric title compound, [Cd DOI: 10.1107/S1600536808025889/ng2485Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Both P atoms have a trigonal\u2013pyramidal conformation. There is a short intra\u00admolecular C\u2014H\u22efP contact with an H\u22efP distance of 2.56\u2005\u00c5. The hydr\u00adoxy group is involved in an intra\u00admolecular O\u2014H\u22ef\u03c0phen\u00adyl inter\u00adaction. The crystal packing shows five very weak inter\u00admolecular C\u2014H\u22ef\u03c0 contacts, with H\u22efCg distances between 3.26 and 3.39\u2005\u00c5 (Cg is the centroid of a phenyl or cyclo\u00adpenta\u00addienyl ring).The absolute configuration of the title compound, [Fe(C DOI: 10.1107/S1600536808038294/nc2123Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular C\u2014H\u22efO hydrogen bonds link neighbouring mol\u00adecules into a one-dimensional supra\u00admolecular structure with an R22(14) ring motif. This structure is further stabilized by \u03c0\u2013\u03c0 stacking inter\u00adactions between adjacent benzene rings [centroid\u2013centroid distance = 3.862\u2005(1)\u2005\u00c5].In the title centrosymmetric mononuclear copper(II) complex, [Cu(C DOI: 10.1107/S1600536809047989/hy2253Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "Inter\u00admolecular C\u2014H\u22efO hydrogen bonds form an eight-membered R 2 2(8) motif. The dihedral angle betwen two benzene rings is 36.34\u2005(9)\u00b0. There are inter\u00admolecular Cu\u22efBr [3.4566\u2005(5)\u2005\u00c5] and Cu\u22ef\u00b7N [3.569\u2005(3)\u2005\u00c5] contacts, which are significantly shorter than the sum of van der Waals radii of the relevant atoms. These inter\u00adactions, along with the inter\u00admolecular C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 [centroid\u2013centroid distances of 3.709\u2005(1) and 3.968\u2005(2)\u2005\u00c5] inter\u00adactions, link neighbouring mol\u00adecules into a one-dimensional infinite chain along the c axis.In the title compound, [Cu(C DOI: 10.1107/S1600536808036635/hy2161Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The geometries of the protonated amino\u00adpyridinium cations reveal amine\u2013imine tautomerism. The crystal packing is influenced by N\u2014H\u22efCl and C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distances = 3.635\u2005(4) and 3.642\u2005(4)\u00b0].The title compound, (C DOI: 10.1107/S160053680904923X/kp2235Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The dihedral angle between the mean planes of the two aromatic rings is 6.16\u2005(6)\u00b0. In the crystal, pairs of inter\u00admolecular weak C\u2014H\u22efO hydrogen bonds link neighboring mol\u00adecules into a chain along the a axis. The crystal structure is further stabilized by two inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances of 3.7252\u2005(13) and 3.8323\u2005(13)\u2005\u00c5.In the title Schiff base complex, [Ni(C DOI: 10.1107/S1600536810007373/bq2198Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two benzene rings are individually planar and make a dihedral angle of 53.90\u00b0. The crystal structure is stabilized by inter\u00admolecular N\u2014H\u22efN hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions (centroid\u2013centroid distance = 3.962\u2005\u00c5).In the mol\u00adecule of the title compound, C DOI: 10.1107/S1600536809013415/rk2138Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit comprises one complex mol\u00adecule and a water mol\u00adecule of crystallization. The water H atoms form bifurcated O\u2014H\u22ef inter\u00admolecular hydrogen bonds with the O atoms of the phenolate and eth\u00adoxy groups with 12(5)R and 12(6)R ring motifs, which may, in part, influence the mol\u00adecular configuration. The dihedral angle between the two O\u2014Cu\u2014N coordination planes is 31.02\u2005(6)\u00b0 and the dihedral angle between the two benzene rings is 34.98\u2005(7)\u00b0. In the crystal structure, mol\u00adecules are linked together by inter\u00admolecular C\u2014H\u22efO inter\u00adactions, forming extended chains along the a axis. The crystal structure is further stabilized by inter\u00admolecular C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 [centroid\u2013centroid = 3.5068\u2005(13)\u2005\u00c5] inter\u00adactions.In the title complex, [Cu(C DOI: 10.1107/S1600536809012859/kj2122Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by C\u2014H\u22efCl inter\u00adactions. There is inter\u00admolecular \u03c0\u2013\u03c0 stacking between adjacent phenanthroline ligands, with a centroid\u2013centroid distance of 3.151\u2005(3)\u2005\u00c5.In the crystal structure of the title complex, [Zn DOI: 10.1107/S1600536809014482/at2764Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The O atoms of the perchlorate anions are disordered with occupancies in the ratio 0.593\u2005(10):0.407\u2005(10). In the crystal, mol\u00adecules are stabilized by two N\u2014H\u22efO hydrogen bonds, forming zigzag chains along the a axis, which are further inter\u00adconnected by N\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.50\u2005(1)\u2005\u00c5] into a three-dimensional network.In the cation of the title compound, [Mn(C DOI: 10.1107/S1600536809029195/tk2501Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CoII atom is located on an inversion center and is coordinated by six O atoms from two water mol\u00adecules and four \u03bc2-carboxyl\u00adate groups of 4-fluoro\u00adbenzoate anions, forming a distorted CoO6 octa\u00adhedron, with Co\u2014O bond lengths in the range 2.071\u2005(2)\u20132.130\u2005(2)\u2005\u00c5. All adjacent O\u2014Co\u2014O angles are in the range 84.78\u2005(6)\u201395.22\u2005(6)\u00b0 and opposite angles are 180.0\u00b0. Each \u03bc-carboxyl\u00adate group of the 4-fluoro\u00adbenzoate anions bridges two symmetry-related CoII atoms. Hydrogen-bonding inter\u00adactions of the coordinated water mol\u00adecules further connect the cobalt\u2013carboxyl\u00adate units, forming layers perpendicular to the a axis. The cobalt\u2013oxygen layers are encased in a sandwich-like fashion by layers of \u03c0-stacked 4-fluoro\u00adbenzoate anions. Within these layers the benzene rings of the 4-fluoro\u00adbenzoate anions are \u03c0-stacked, with centroid\u2013centroid distances of 3.432\u2005(4)\u2005\u00c5.The hydro\u00adthermal reaction of CoCO DOI: 10.1107/S1600536809014913/zl2189Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal packing exhibits \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings from neighbouring mol\u00adecules [centroid\u2013centroid distance = 3.616\u2005(4)\u2005\u00c5] and weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions.In the title mol\u00adecule, C DOI: 10.1107/S1600536808000895/cv2379Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure exhibits four inter\u00admolecular non-classical C\u2014H\u22efO hydrogen bonds. In addition, the crystal structure contains aromatic \u03c0\u2013\u03c0 inter\u00adactions between the furan and benzene rings of adjacent mol\u00adecules [centroid\u2013centroid distance = 3.743\u2005(2)\u2005\u00c5], and two inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, C DOI: 10.1107/S1600536809025938/ez2175Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, the complex mol\u00adecules are linked into a three-dimensional network by inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, and \u03c0\u2013\u03c0 stacking inter\u00adactions with centroid\u2013centroid separations of 3.758\u2005(2) and 3.597\u2005(1)\u2005\u00c5.In the title compound, [Zn(C DOI: 10.1107/S1600536809009180/hy2185Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Two N atoms and the Cl atom are in equatorial positions while the remaining two N atoms occupy apical sites, the equatorial Cu\u2014N bonds being significantly longer than the two apical Cu\u2014N bonds. The N=C\u2014O\u2014C torsion angles involving the four eth\u00adoxy groups are in the range 161.5\u2005(8) to 177.0\u2005(5)\u00b0. In the crystal structure, there are significant \u03c0\u2013\u03c0 stacking inter\u00adactions between inversion-related rings of phenanthroline groups with centroid\u2013centroid distances in the range 3.649\u2005(4)\u20133.790\u2005(4)\u2005\u00c5.In the title complex, [CuCl(C DOI: 10.1107/S160053680804138X/lh2739Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Weak \u03c0\u2013\u03c0 stacking inter\u00adactions, with centroid\u2013centroid distances of 3.862\u2005(2) and 3.887\u2005(5)\u2005\u00c5, and significant C\u2014H\u22ef\u03c0 inter\u00adactions help to stabilize the crystal structure. The atoms of the unique terminal 4-pyridine\u00adpropane group are disordered over two sites, the ratio of refined occpancies being 0.712\u2005(7):0.288\u2005(7).In the title compound, [Mn(C DOI: 10.1107/S1600536809009891/lh2780Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There are two formula units in the asymmetric unit. Molecules are further assembled into a three-dimensional network through C\u2014H\u22efCl contacts, a Cu\u22efCl weak inter\u00adaction [3.161\u2005(2)\u2005\u00c5], O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds. The three water mol\u00adecules of the asymmetric unit are distributed over five positions with one full and two approximately half occupancies, while a tyrosine side chain in one of the complex mol\u00adecules is disordered over two positions [occupancies 0.507\u2005(5) and 0.493\u2005(5)].In the title compound, [Cu(C DOI: 10.1107/S1600536808012038/cs2072Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordinated water mol\u00adecule acting as the bridging ligand is located on a twofold axes and the complex mol\u00adecule displays C 2 mol\u00adecular symmetry. The Fe\u22efFe separation in the binuclear complex is 3.490\u2005(3)\u2005\u00c5. The crystal structure is stabilized by hydrogen bonding and \u03c0\u2013\u03c0 stacking inter\u00adactions .In the title binuclear complex, [Fe DOI: 10.1107/S1600536808014207/kp2167Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The three pyridine rings are approximately coplanar, with a maximum deviation of 0.03\u2005\u00c5 from the mean plane. The phen\u00adoxy substituent makes a dihedral angle of 18.1\u2005(2)\u00b0 with the central pyridine ring. The benzyl group has a C\u2014O\u2014C\u2014C torsion angle of 77.62\u2005(8)\u00b0 relative to the phen\u00adoxy ring. In the crystal, mol\u00adecules are linked via C\u2014H\u22efO hydrogen bonds.In the title compound, [Zn(CH DOI: 10.1107/S1600536809049502/wm2284Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "II ion in the title compound, [Zn(C15H14N10)(H2O)2](ClO4)2, lies on a centre of symmetry. The distorted N4O2 octa\u00adhedral coordination environment around the Zn atom is composed of two 1,3-bis\u00ad[5-(2-pyrid\u00adyl)-2H-tetra\u00adzol-2-yl]propane ligands (L1) and two water mol\u00adecules, coordinated in trans positions. The ligand acts as a typical bidentate chelating ligand through one of its 2-pyridyl-2H-tetra\u00adzole units, forming a five-membered Zn\u2014N\u2014C\u2014C\u2014N metallacycle with a small N\u2014Zn\u2014N bite angle [77.40\u2005(8)\u00b0]. The other 2-pyridyl-2H-tetra\u00adzole unit remains uncoordinated. The average Zn\u2014N distance (2.156\u2005\u00c5) is somewhat longer than the distance between the ZnII center and the aqua ligand [2.108\u2005(2)\u2005\u00c5]. The coordinated pyrid\u00adyl-tetra\u00adzoyl rings are quasi-coplanar, making a dihedral angle of 1.9\u2005(2)\u00b0, while the uncoordinated rings show a larger inter\u00adplanar angle of 21.3\u2005(2)\u00b0. The flexible propane spacer displays a zigzag chain. Inter\u00admolecular O\u2014H\u22efN and O\u2014H\u22efO inter\u00adactions result in two-dimensional polymeric structures parallel to (100). Two C atoms of the spacer are disordered over two positions, with site occupancy factors of ca 0.85 and 0.15.The Zn DOI: 10.1107/S1600536808006703/bg2168Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal, mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds into chains running along [010]. Adjacent chains are joined together by weak \u03c0\u2013\u03c0 inter\u00adactions between benzene rings [centroid\u2013centroid distance = 4.040\u2005(2)\u2005\u00c5].There are two mol\u00adecules in the asymmetric unit of the title compound, C DOI: 10.1107/S1600536810000139/fk2009Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "IV atom in the title compound, [Sn(C6H5)2(C18H11ClN2O3)], is O,N,O\u2032-chelated by the deprotonated Schiff base ligand and further bonded by two phenyl rings in a distorted cis-C2SnNO2 trigonal-bipyramidal geometry [C\u2014Sn\u2014C = 125.7\u2005(2)\u00b0]. The two phenyl rings are oriented at a dihedral angle of 55.2\u2005(3)\u00b0. Intra\u00admolecular O\u2014H\u22efN hydrogen bonding is present in the crystal structure.The Sn DOI: 10.1107/S1600536809050107/xu2687Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The crystal structure is stabilized by C\u2014H\u22efBr hydrogen bonds and inter\u00admolecular C\u2014Br\u22ef\u03c0 inter\u00adactions [C\u22ef\u03c0 = 3.57\u2005(1)\u2005\u00c5].In the title compound, [Sb(C DOI: 10.1107/S1600536808033783/lx2073Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "An intra\u00admolecular O\u2014H\u22efO hydrogen bond occurs. In the crystal, inter\u00admolecular O\u2014H\u22efO hydrogen bonds link the mononuclear complexes into chains extending parallel to [010]. Furthermore, \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.5696\u2005(6)\u2005\u00c5] stabilize the crystal structuure. In the mononuclear title complex, [Ni(C DOI: 10.1107/S1600536810034252/bt5331Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There is an intra\u00admolecular O\u2014H\u22efO hydrogen bond. Inter\u00admolecular C\u2014H\u22efO hydrogen bonds result in the formation of a three-dimensional network and \u03c0\u2013\u03c0 stacking inter\u00adactions [3.44\u20133.83\u2005\u00c5] are observed between symmetry-related rings of 2-methyl\u00adfuropyridine. Further inter\u00adactions in the crystal structure are a short Cl\u22efCl inter\u00adaction [3.384\u2005(2)\u00c5] and C\u2014H\u22ef\u03c0 inter\u00adactions between 2-methyl\u00adfuropyridine rings.In the title compound, [Cu(C DOI: 10.1107/S1600536808008404/om2220Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There are significant differences between chemically equivalent coordination bond lengths. The crystal structure is stabilized by weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds and weak \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance 3.495\u2005(1)\u2005\u00c5]. In one of the anions one nitro group is rotationally disordered about the C\u2014N bond with refined occupancies of 0.524\u2005(8) and 0.476\u2005(8).In the title complex, [Co(C DOI: 10.1107/S1600536809035934/lh2889Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular O\u2014H\u22efO hydrogen-bonding inter\u00adactions lead to infinite chains, which are further self-assembled into a supra\u00admolecular network through inter\u00admolecular N\u2014H\u22efO hydrogen-bonding inter\u00adactions and \u03c0\u2013\u03c0 stacking [centroid\u2013centroid distance = 3.717\u2005(2)\u2005\u00c5].In the mononuclear title compound, [Ni(C DOI: 10.1107/S1600536808009471/ng2442Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The central 1,6-diazecine ring adopts the common chair conformation invariably found in the family of complexes bearing such ligands. The CuII atoms have an octa\u00adhedral geometry, with a very strong tetra\u00adgonal distortion due to the Jahn\u2013Teller effect. Axial sites are occupied by a nitrate ion and a water mol\u00adecule. The Cu\u22efCu separations [7.3580\u2005(9) and 7.3341\u2005(9)\u2005\u00c5] are compatible with a potential catecholase activity. Neighboring mol\u00adecules in the crystal structure are connected via O\u2014H\u22efO hydrogen bonds formed by water mol\u00adecules and carboxyl\u00adate O atoms. N\u2014H\u22efO hydrogen bonds are also present.The title compound, [Cu II complexes based on related bis\u00ad(amino\u00adimidazole) ligands, which were designed as models of the catechol oxidaze active site, see: Driessen et al. (NO3)2(H2O)4]\u00b73H2O = 0.030 wR(F 2) = 0.082 S = 1.02 6396 reflections433 parameters22 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.50 e \u00c5\u22123 \u0394\u03c1min = \u22120.48 e \u00c5\u22123 \u0394\u03c1 XSCANS used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008Mercury (Macrae et al., 2006SHELXL97.Data collection: 10.1107/S1600536808023969/hy2146sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536808023969/hy2146Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The C\u2014Al\u2014C angles range from 113.25\u2005(7) to 116.27\u2005(8)\u00b0, much larger than the O\u2014Al\u2014C angles, which range from 103.39\u2005(7) to 103.90\u2005(6)\u00b0. The tetra\u00adhydro\u00adfuran ring adopts an envelope conformation. The crystal packing is stabilized by C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, [Al(C DOI: 10.1107/S1600536808032091/ci2683Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit contains one PbII ion, two benzoate ligands and one water mol\u00adecule. The Pb\u2014O bond distances are in the range 2.494\u2005(4)\u20132.735\u2005(4)\u2005\u00c5. The Pb\u22efPb distance is 4.0683\u2005(4)\u2005\u00c5, indicating an insignificant metal\u2013metal inter\u00adaction. The PbII atom has a distorted penta\u00adgonal-bipyramidal geometry chelated by two carboxyl\u00adate O atoms. The Pb atoms are bridged through a coordinating water mol\u00adecule and two carboxyl\u00adate O atoms from another two benzoate ligands, giving an infinite three-dimensional supra\u00admolecular structure. O\u2014H\u22efO hydrogen-bonding inter\u00adactions involved the coordinating water and carboxyl\u00adate O atoms enhance the stability of the supra\u00admolecular arrangement.The reaction of lead(II) nitrate and benzoic acid in aqueous solution yields the title polymer, [Pb(C For rel al. 2007. 7H5O2)2(H2O)] = 0.034 wR(F 2) = 0.087 S = 1.09 2664 reflections181 parametersH-atom parameters constrainedmax = 3.35 e \u00c5\u22123 \u0394\u03c1min = \u22121.31 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008SHELXTL (Sheldrick, 2008SHELXTL.Data collection: 10.1107/S1600536809028542/fj2238sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536809028542/fj2238Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecules are linked into chains running parallel to the b axis by inter\u00admolecular N\u2014H\u22efO hydrogen bonds and by \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.669\u2005(2)\u2005\u00c5] involving centrosymmetrically related imidazole rings.In the title mononuclear complex, [Cu(C DOI: 10.1107/S1600536808012440/rz2208Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "A three-dimensional network is formed via inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the imidazole and benzene rings of neighboring mol\u00adecules [centroid\u2013centroid distance = 3.856\u2005(2)\u2005\u00c5].In the title mononuclear complex, [Ni(C DOI: 10.1107/S1600536809005960/hy2181Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The strongest cation\u2013cation O\u2014H\u22efO hydrogen bond in the structure, together with other strong cation\u2013cation N\u2014H\u22efO hydrogen bonds, generates a succession of infinite chains of R 2 2(8) rings along the b axis. Additional cation\u2013cation C\u2014H\u22efO hydrogen bonds link these chains into two-dimensional layers formed by alternating R 4 4(24) and R 4 2(12) rings. Connections between these layers are provided by the strong cation\u2013anion N\u2014H\u22efO hydrogen bonds, as well as by one weak C\u2014H\u22efO inter\u00adaction, thus forming a three-dimensional network. Some of the cation\u2013anion N\u2014H\u22efO hydrogen bonds are bifurcated of the type D\u2014H\u22ef.In the title compound, C DOI: 10.1107/S1600536809031730/fb2157Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Au atom exists within a linear geometry defined by an S,P-donor set with a deviation from linearity [S\u2014Au\u2014P = 176.86\u2005(6)\u00b0] due to the close approach of the thio\u00adcarbamate O atom [Au\u22efO = 3.108\u2005(5)\u2005\u00c5]. The mol\u00adecule has a U-shaped geometry which facilitates the formation of an intra\u00admolecular Au\u22efAu inter\u00adaction [3.0231\u2005(5)\u2005\u00c5]. In the crystal, the presence of C\u2014H\u22efOnitro contacts leads to the formation of layers with substantial voids; these are occupied by the solvent mol\u00adecules of crystallization, which are held in place by C\u2014H\u22efS contacts. The dinuclear title mol\u00adecule, [Au For the al. 1993. 2Fe(C10H11N2O3S)2(C17H14P)2]\u00b72CHCl3 = 0.045 wR(F 2) = 0.104 S = 0.97 7148 reflections357 parametersH-atom parameters constrainedmax = 2.08 e \u00c5\u22123 \u0394\u03c1min = \u22120.74 e \u00c5\u22123 \u0394\u03c1 SMART used to solve structure: PATTY in DIRDIF92 (Beurskens et al., 1992SHELXL97 (Sheldrick, 2008DIAMOND (Brandenburg, 2006SHELXL97.Data collection: 10.1107/S1600536809043864/hb5168sup1.cif Crystal structure: contains datablocks global, I. DOI: 10.1107/S1600536809043864/hb5168Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Pt atom has an octa\u00adhedral coordination. In the crystal structure, inter\u00admolecular N\u2014H\u22efCl and C\u2014H\u22efCl hydrogen bonds result in the formation of a supra\u00admolecular structure. There is a \u03c0\u2013\u03c0 contact between the pyridine rings [centroid\u2013centroid distance = 4.235\u2005(1)\u2005\u00c5].The asymmetric unit of the title compound, (C DOI: 10.1107/S1600536808025257/hk2508Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angles between the carboxyl\u00adate groups and the adjacent benzene rings are 11.33\u2005(13) and 10.90\u2005(9)\u00b0 and the benzene rings are oriented at a dihedral angle of 67.88\u2005(6)\u00b0. The uncoordinated water mol\u00adecules link the carboxyl\u00adate groups and coordinated water mol\u00adecules via O\u2014H\u22efO hydrogen bonding. Weak C\u2014H\u22ef\u03c0 inter\u00adactions are also found in the crystal structure.In the crystal structure of the title complex, {[Mn(C DOI: 10.1107/S1600536809021060/xu2525Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The equatorial positions are occupied by two N atoms from a 1,10-phenanthroline ligand [Cu\u2014N = 1.994\u2005(3) and 2.027\u2005(3)\u2005\u00c5] and two O atoms from dichloro\u00adacetate ligands and a water mol\u00adecule [Cu\u2014O = 1.971\u2005(2) and 1.939\u2005(2)\u2005\u00c5]. One O atom from another dichloro\u00adacetate ligand occupies the apical positon [Cu\u2014O = 2.152\u2005(3)\u2005\u00c5]. Inter\u00admolecular O\u2014H\u22efO hydrogen bonds link the mol\u00adecules into centrosymmetric dimers. The crystal packing also exhibits weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds, \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.734\u2005(2)\u2005\u00c5] and short inter\u00admolecular Cl\u22efCl contacts [3.306\u2005(2) and 3.278\u2005(2)\u2005\u00c5].In the title complex, [Cu(C DOI: 10.1107/S1600536808042578/cv2493Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by aromatic \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of neighbouring mol\u00adecules [centroid\u2013centroid distance = 3.814\u2005(9)\u2005\u00c5], and possibly by weak C\u2014H\u22ef\u03c0 inter\u00adactions. In addition, the crystal structure exhibits three inter\u00admolecular C\u2014H\u22efO non-classical hydrogen bonds. The ethyl group bonded to carboxyl\u00adate O atom is disordered over two positions, with refined site-occupancy factors of 0.686\u2005(18) and 0.314\u2005(18).The title compound, C DOI: 10.1107/S1600536809009775/rk2134Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecular packing in the crystal is stabilized by inter\u00admolecular N\u2014H\u22efO inter\u00adactions, linking the mol\u00adecules into infinite chains along the c axis. In addition, there are weak C\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, C DOI: 10.1107/S1600536810009803/bt5217Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The thia\u00adzole ring forms dihedral angles of 83.7\u2005(2) and 34.4\u2005(2)\u00b0 with the benzene ring of the benzodioxole ring and the fused phenyl ring, respectively. The mol\u00adecular conformation is stabilized by an intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction. The crystal packing features inter\u00admolecular C\u2014H\u22efN, C\u2014H\u22efO hydrogen bonds and weak C\u2014H\u22ef\u03c0 inter\u00adactions.In the title compound, [Fe(C DOI: 10.1107/S160053681003638X/rz2480Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There is a \u03c0\u2013\u03c0 contact between the pyridine rings [centroid\u2013centroid distance = 3.775\u2005(6)\u2005\u00c5].In the mol\u00adecule of the title compound, [HgI DOI: 10.1107/S1600536808028791/hk2525Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is assembled via hydrogen-bonding inter\u00adactions of two kinds, N(pyridine/amine)\u2014H\u22efBr\u2014Sn, along with C\u2014Br\u22efBr\u2014Sn interactions [3.4925\u2005(19)\u2005\u00c5]. The cations are involved in \u03c0\u2013\u03c0 stacking, which adds an extra supra\u00admolecularity as it presents a strong case of offset-face-to-face motifs [centroid\u2013centroid distance = 3.577\u2005(3)\u2005\u00c5]. The inter\u00admolecular hydrogen bonds, short Br\u22efBr inter\u00adactions and \u03c0\u2013\u03c0 stacking result in the formation of a three-dimensional supra\u00admolecular architecture.The asymmetric unit of the title compound, (C DOI: 10.1107/S1600536809015189/hk2669Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "L) and the title silver(I) complex, namely bis\u00ad[\u03bc-1--2-naphthol]bis\u00ad{[1--2-naphthol]silver(I)} dinitrate monohydrate, [Ag2(C13H10N4O)4](NO3)2\u00b7H2O, were synthesized. Each silver center in the dimeric complex (related by an inversion centre) is coordinated by two bridging L ligands and one additional L ligand in a monodentate fashion, exhibiting a distorted trigonal-planar coordination. The discrete dimers are further linked through O\u2014H\u22efO hydrogen bonds and weak \u03c0\u2013\u03c0 stacking inter\u00adactions [the shortest atom\u2013atom separation is ca 3.46\u2005\u00c5 between the parallel stacking pairs]. Intramolecular O\u2014H\u22efN hydrogen bonds are also present.The new ligand 1--2-naphthol ( DOI: 10.1107/S1600536809004760/wk2097Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "This compound is composed of an anionic complex, [Al(pydc)2]\u2212, a protonated 2,2\u2032-bipyridine mol\u00adecule as a counter-ion, +, and three uncoordinated water mol\u00adecules. The anion is a six-coordinate complex, with the AlIII atom in a distorted octa\u00adhedral geometry coordinated by two tridentate pyridine-2,6-dicarboxyl\u00adate groups. In the crystal structure, inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO, N\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds, \u03c0\u2013\u03c0 stacking between two aromatic rings [centroid\u2013centroid distance = 3.827\u2005(10)\u2005\u00c5], and C=O\u22ef\u03c0 stacking , connect the various components to form a supra\u00admolecular structure.The title compound, (C DOI: 10.1107/S1600536808015973/su2057Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "IV compound, [Sn4(C7H6Cl)8Cl2O2(OH)2], has site symmetry 2\u2212 and two OH\u2212 anions bridge four SnIV cations to form the tetra\u00adnuclear compound. The two independent SnIV cations assume SnO3C2 and SnO2C2Cl distorted trigonal-bipyramidal coordination geometries. Intra\u00admolecular O\u2014H\u22efCl hydrogen bonding is present in the structure. One Cl atom of a chloro\u00adbenzyl ligand is disordered over two sites with an occupancy ratio of 0.693\u2005(2):0.307\u2005(2).The title tetra\u00adnuclear Sn DOI: 10.1107/S1600536809045176/xu2652Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, mol\u00adecules are linked into centrosymmetric dimers by pairs of O\u2014H\u22efO hydrogen bonds, and the dimers are linked together by \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.8310\u2005(13)\u2005\u00c5] and C\u2014H\u22efO bonds.In the mol\u00adecule of the title compound, C DOI: 10.1107/S1600536809049721/fk2006Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The bond distances for Sn\u2014C, Sn\u2014N and Sn\u2014O are in the ranges 2.097\u2005(3)\u20132.098\u2005(3), 2.298\u2005(2)\u20132.623\u2005(2) and 2.157\u2005(2)\u20132.266\u2005(2)\u2005\u00c5, respectively. The mol\u00adecular structure of the monomeric compound is stabilized by three intra\u00admolecular C\u2014H\u22efO hydrogen bonds, all involving bipyridine C\u2014H groups.In the crystal structure of the title compound, [Sn(CH DOI: 10.1107/S1600536808006090/im2056Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecular structure features intra\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds. In the crystal structure, the complex mol\u00adecules are assembled into a two-dimensional supra\u00admolecular layer parallel to (011) via O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridyl and pyrazole rings [centroid\u2013centroid distances = 3.544\u2005(2) and 3.722\u2005(3)\u2005\u00c5].In the mononuclear title complex, [Cd(C DOI: 10.1107/S1600536810034616/hy2347Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There is a \u03c0\u2013\u03c0 stacking inter\u00adaction between the symmetry-related 1,10-phenanthroline ligands with a centroid\u2013centroid distance of 3.5578\u2005(16)\u2005\u00c5 and a perpendicular distance of 3.445\u2005(su?)\u2005\u00c5 between the relevant rings.In the title complex, [CdCl DOI: 10.1107/S1600536809032358/bt5029Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Two AgI ions are each bonded to two N atoms from two isonicotinate ligands in a linear or bow-like fashion [N\u2014Ag\u2014N angles = 178.6\u2005(2) and 147.1\u2005(2)\u00b0]. These metal ions are connected by the isonicotin\u00adate ligands into a layer parallel to (010). O\u2014H\u22efO hydrogen bonds donated by the coordinated and uncoordinated water mol\u00adecules and intra\u00adlayer \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings [centroid\u2013centroid distances = 3.551\u2005(4) and 3.555\u2005(4)\u2005\u00c5] are observed. The layers inter\u00adact with each other by inter\u00adlayer Ag\u22efO(aqua) contacts [2.731\u2005(4)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings [centroid\u2013centroid distances = 3.466\u2005(3) and 3.516\u2005(3)\u2005\u00c5], resulting in the formation of a three-dimensional supra\u00admolecular structure.In the title compound, {[Ag DOI: 10.1107/S1600536810035634/hy2346Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral angle between the planes of the phenanthridine ring systems is 69.92\u2005(3)\u00b0. An intra\u00admolecular C\u2014H\u22efCl inter\u00adaction results in the formation of a planar five-membered ring, which is oriented at a dihedral angle of 8.32\u2005(3)\u00b0 with respect to the adjacent phenanthridine ring system. In the crystal structure, \u03c0\u2013\u03c0 contacts between the phenanthridine systems may stabilize the structure. Two weak C\u2014H\u22ef\u03c0 inter\u00adactions are also found.In the mol\u00adecule of the title compound, [ZnCl DOI: 10.1107/S160053680901959X/hk2696Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The complex anion is approximately planar with a maximum deviation of 0.097\u2005(1)\u2005\u00c5. In the 1-(4-nitro\u00adbenz\u00adyl)-4-cyano\u00adpyridinium cation, the pyridine ring is twisted at a dihedral angle of 73.84\u2005(16)\u00b0 with respect to the benzene ring. \u03c0-\u03c0 stacking is observed between nearly parallel [dihedral angle = 4.71\u2005(7)\u00b0] dithiole and benzene rings, the centroid\u2013centroid distance being 3.791\u2005(2)\u2005\u00c5.In the title salt, (C DOI: 10.1107/S1600536810037426/xu5018Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "O\u2014H\u22efO and O\u2014H\u22efCl hydrogen bonds involving hydr\u00adoxy groups and one of coordinated Cl atoms link complexes in the crystal packing. There is a \u03c0\u2013\u03c0 stacking inter\u00adaction between adjacent 1,10-phenanthroline rings, with a distance of 3.675\u2005(2)\u2005\u00c5 between the centroids of the pyridine and benzene rings.In the title complex, [CdCl DOI: 10.1107/S1600536809037465/kp2226Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The RuII ion is chelated by a 10,11,12,13-tetra\u00adhydro\u00addipyridophenazine ligand and two 2,9-dimethyl-1,10-phenanthroline ligands in a distorted octa\u00adhedral geometry. The two uncoord\u00adinated water mol\u00adecules are disordered over five positions, with an occupancy factor of about 0.4 for each site. A supra\u00admolecular structure is formed by weak \u03c0\u2013\u03c0 inter\u00adactions between neighbouring mol\u00adecules, with centroid\u2013centroid distances of 3.618\u2005(2) and 3.749\u2005(2)\u2005\u00c5.The title compound, [Ru(C DOI: 10.1107/S1600536810002874/hy2273Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The combination of coordinative bonds, O\u2014H\u22efN and O\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.630\u2005(2)\u2005\u00c5] results in the stabilization of a supra\u00admolecular structure. All uncoordinated water molecules are disordered. Thermogravimetric analysis reveals that the title complex loses the four crystal water mol\u00adecules at about 333\u2005K, then the anhydrous phase loses no further mass up to about 573\u2005K, above which decomposition occurs.The hydro\u00adthermal reaction of CoCl DOI: 10.1107/S1600536809030165/at2833Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The complex cations are linked by hydrogen bonds between the carboxyl groups into a chain. The chains are further connected through C\u2014H\u22efO hydrogen bonds and a weak Ag\u22efO inter\u00adaction [2.757\u2005(8)\u2005\u00c5] into a layer. Another Ag\u22efO inter\u00adaction [2.899\u2005(2)\u2005\u00c5] and a C\u2014H\u22efO hydrogen bond connect the layers into a three-dimensional network.In the title compound, [Ag(C DOI: 10.1107/S1600536808044206/hy2176Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CoII center is coordinated by six H2O mol\u00adecules in a distorted octa\u00adhedral coordination environment. In the crystal structure, intra- and inter\u00admolecular O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds link the cations and anions into a three-dimensional network. \u03c0\u2013\u03c0 contacts between the tetra\u00adzole rings [centroid\u2013centroid distance = 3.346\u2005(1)\u2005\u00c5] may further stabilize the structure.The asymmetric unit of the title complex, [Co(H DOI: 10.1107/S1600536809028463/hk2740Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The bridging carbonyl C\u2014C(=O)\u2014C plane makes dihedral angles of 45.55\u2005(6) and 28.62\u2005(7)\u00b0, respectively, with the naphthalene ring system and the benzene ring. Weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions stabilize the crystal packing.In the title compound, C DOI: 10.1107/S1600536810006859/bt5198Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The centrosymmetric complex cation involves intra\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.862\u2005(4)\u2005\u00c5] between the central pyridine and benzene rings. In the crystal structure, inter\u00admolecular C\u2014H\u22efO hydrogen bonds result in the formation of a supra\u00admolecular network.In the binuclear title complex, [Ag DOI: 10.1107/S1600536809046832/hy2238Isup2.hklStructure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "The Ni\u2014N bond distances range from 2.1061\u2005(18) to 2.1425\u2005(18)\u2005\u00c5. The Mo\u2014C and C\u2014N distances in the [Mo(CN)8] unit range from 2.154\u2005(2) to 2.174\u2005(2)\u2005\u00c5 and 1.149\u2005(3) to 1.156\u2005(3)\u2005\u00c5, respectively. The complex ions and water mol\u00adecules are linked by weak N\u2014H\u22efN/O and O\u2014H\u22efN/O hydrogen bonds into a three-demensional structure.In the title compound, [Ni(C DOI: 10.1107/S1600536809051964/pv2243Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two N atoms and two O atoms around the CuII atom are trans to each other, with an O\u2014Cu\u2014O bond angle of 177.00\u2005(9)\u00b0 and an N\u2014Cu\u2014N bond angle of 165.63\u2005(10)\u00b0. The average distances between the CuII atom and the coordinated O and N atoms are 1.905\u2005(2) and 1.995\u2005(2)\u00c5, respectively.In the title complex, [Cu(C DOI: 10.1107/S1600536810037888/rk2236Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II complex, [Ni(C9H10NO2)2(C10H14N2O)2(H2O)2], contains two dimethyl\u00adamino\u00adbenzoate (DMAB), two diethyl\u00adnicotinamide (DENA) ligands and two water mol\u00adecules, all of them monodentate. The four O atoms in the equatorial plane around the NiII atom form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two pyridine N atoms of the DENA ligands in axial positions. The NiII atom is displaced by 0.681\u2005(1)\u2005\u00c5 out of the least-squares plane of the carboxyl\u00adate group. The dihedral angle between the carboxyl\u00adate group and the adjacent benzene ring is 5.61\u2005(7)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 73.20\u2005(4)\u00b0. An intra\u00admolecular O\u2014H\u22efO hydrogen bond results in the formation of a six-membered ring with a twisted conformation. In the crystal structure, inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link mol\u00adecules into a three-dimensional network. Two weak C\u2014H\u22ef\u03c0 inter\u00adactions are also present.The centrosymmetric title Ni N,N-diethyl\u00adnicotinamide, an important respiratory stimulant, see: Bigoli et al. 2(C10H14N2O)2(H2O)2] = 0.029 wR(F 2) = 0.076 S = 1.04 4819 reflections253 parametersH atoms treated by a mixture of independent and constrained refinementmax = 0.47 e \u00c5\u22123 \u0394\u03c1min = \u22120.41 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S1600536809030098/xu2570sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536809030098/xu2570Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Pt\u2014P bond lengths of 2.2536\u2005(8) and 2.2513\u2005(8)\u2005\u00c5 and the Pt\u2014Cl bond lengths of 2.3750\u2005(8) and 2.3588\u2005(8)\u2005\u00c5 are normal. This crystal form is a polymorph of a structure reported previously .The title complex, [PtClasai 1976. Bull. C DOI: 10.1107/S1600536808000603/pv2062Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Neighboring CoII atoms are linked together by two Cl bridges, forming a dinuclear CoII complex with inversion symmetry. There are inter\u00admolecular O\u2014H\u22efCl hydrogen bonds and inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions between adjacent bpy ligands [centroid\u2013centroid distance = 3.617\u2005(2)\u2005\u00c5] in the structure.The title complex, [Co DOI: 10.1107/S1600536809027846/at2843Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The complexes are assembled into a three-dimensional network via C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions. The mean inter\u00adplanar distance between adjacent phenanthroline ligands is 3.399\u2005(2)\u2005\u00c5.In the crystal structure of the title compound, [Zn(C DOI: 10.1107/S1600536808006004/is2271Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CoII cation is coordinated by two Cl\u2212 ions, one N atom from the 5-(4-pyridinio)tetra\u00adzolate zwitterion and three O atoms from three water mol\u00adecules in a distorted octa\u00adhedral geometry. In the crystal, mol\u00adecules are linked into a three-dimensional network by N\u2014H\u22efCl hydrogen bonds and O\u2014H\u22efO/N/Cl hydrogen bonds involv\u00ading both coordinated and uncoordinated water mol\u00adecules. Strong \u03c0\u2013\u03c0 stacking is present between parallel pyridinium and tetra\u00adzolate rings [centroid\u2013centroid distances = 3.411\u2005(2) and 3.436\u2005(2)\u2005\u00c5].The title compound, [CoCl DOI: 10.1107/S1600536809024337/ci2831Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Both the cationic and anionic portions of the starting proton-transfer compound are involved in the complexation. The NiII atom has a distorted octa\u00adhedral geometry and is hexa\u00adcoordinated by three O atoms and three N atoms from one phen fragment (as a bidentate ligand), one (pydc)2\u2212 unit (as a tridentate ligand) and one water mol\u00adecule. In the crystal structure, extensive O\u2014H\u22efO, O\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds with D\u22efA distances ranging from 2.573\u2005(2) to 3.385\u2005(2)\u2005\u00c5, \u03c0\u2013\u03c0 inter\u00adactions between the phen ring systems and inter\u00admolecular C\u2014O\u22ef\u03c0 inter\u00adactions connect the various components together.The title compound, [Ni(C DOI: 10.1107/S1600536808039378/om2263Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The other NiII ion is also six-coordinated, by four other pyridine N atoms from four other amino\u00adpyridine ligands and two carbonate O atoms to complete a distorted octa\u00adhedral geometry. In the crystal structure, mol\u00adecules are linked into an infinite three-dimensional network by O\u2014H\u22efO, N\u2014H\u22efCl, N\u2014H\u22efO, O\u2014H\u22efN, C\u2014H\u22efO, C\u2014H\u22efN and C/N\u2014H\u22ef\u03c0 inter\u00adactions involving the pyridine rings.In the title compound, [Ni DOI: 10.1107/S1600536808033321/at2653Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The apical Cu\u2014O bond [2.3520\u2005(16)\u2005\u00c5] is much longer than the basal Cu\u2014O and Cu\u2014N bonds [1.9139\u2005(14)\u20132.0136\u2005(17)\u2005\u00c5]. The carboxyl\u00adate group bridges CuII atoms, forming a zigzag chain along the a axis.In the title compound, [Cu(C DOI: 10.1107/S1600536809037520/is2450Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The MoVI atom shows a distorted octa\u00adhedral environment, with the phenanthroline N atoms and the two oxide groups forming the equatorial plane and the F atoms occupying the apical positions. Weak C\u2014H\u22efO and C\u2014H\u22efF hydrogen-bonding contacts and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.662\u2005(1)\u2005\u00c5] connect the complex mol\u00adecules into a three-dimensional supra\u00admolecular framework.The title compound, [MoF DOI: 10.1107/S1600536809031626/si2192Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "A short intra\u00admolecular C\u2014H\u22ef\u03c0 interaction is observed involving a substituted \u03b75-C5H4 ring and an ortho H atom of the benzene ring on the opposite side of the mol\u00adecule. In the crystal structure, there are no classical hydrogen bonds: inter\u00adactions comprise a short C6\u2014H\u22ef\u03c0(C6) inter\u00adaction involving substituted benzene rings and two C\u2014H\u22efO=C inter\u00adactions per mol\u00adecule.The title compound, [Fe DOI: 10.1107/S1600536809006278/bg2237Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials: interactive version of Fig. 2 Enhanced figure:"} +{"text": "There is a mean deviation of 0.016\u2005(4)\u2005\u00c5 from the least-squares plane defined by the nine constituent benzofuran atoms. The crystal structure is stabilized by aromatic \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of neighbouring mol\u00adecules [centroid\u2013centroid distance = 3.689\u2005(7)\u2005\u00c5]and by a weak C\u2014H\u22ef\u03c0 interaction between an H atom of the methylene group bonded to the carboxylate O atom and the benzene ring of an adjacent molecule. In addition, the crystal structure exhibits weak non-classical inter\u00admolecular C\u2014H\u22efO hydrogen bonds. The chloro\u00adethyl group is disordered over two positions, with refined site-occupancy factors of 0.767\u2005(6) and 0.233\u2005(6).In the title compound, C DOI: 10.1107/S1600536809012847/sj2602Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular C\u2014H\u22efCl hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pyridine rings [centroid\u2013centroid distances = 3.788\u2005(1) and 3.957\u2005(1)\u2005\u00c5] are present in the crystal structure.In the title compound, [CoCl DOI: 10.1107/S1600536810036846/hy2353Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The terpyridyl motif in each fctpy ligand is coplanar, but the cyclo\u00adpenta\u00addienyl ring is twisted by 9.5\u2005(2)\u00b0 out of coplanarity with each central pyridine. The two cyclo\u00adpenta\u00addienyl rings of the ferrocenyl group are almost eclipsed with a deviation of 4.5\u2005(1)\u00b0. In addition, inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance 3.753\u2005(2)\u2005\u00c5] are present between the cyclo\u00adpenta\u00addienyl and outer pyridyl rings of the fctpy ligands. One of the perchlorate anions is equally disordered over two positions.In the title complex, [FeZn(C DOI: 10.1107/S1600536809023939/jh2081Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "It differs from the first ortho\u00adrhom\u00adbic polymorph [Pan, Lin & Zheng ligand. In the present structure, the Cu atom is shifted from the mean plane of the dmp ligand by only 0.005\u2005(1)\u2005\u00c5, compared with 0.318\u2005(6)\u2005\u00c5 in the ortho\u00adrhom\u00adbic form. Hydrogen-bonding and \u03c0\u2013\u03c0 stacking inter\u00adactions (mean inter\u00adplanar distance of 3.59\u2005\u00c5 in the title compound) in the two different polymorphs are both essential to the supra\u00admolecular assembly.A new monoclinic polymorphic form of the title compound, [Cu(HCOheng 2005. Z. Kris DOI: 10.1107/S1600536808022812/fj2128Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00admolecular O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonding, as well as \u03c0\u2013\u03c0 stacking between parallel thia\u00adzole rings [centroid\u2013centroid distance 3.531\u2005(8)\u2005\u00c5], helps to stabilize the crystal structure.In the crystal structure of the title compound, [Ni(C DOI: 10.1107/S1600536809017978/xu2518Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Together the ligands create a slightly distorted square-planar cordination environment for the Rh(I) atom. There are three mol\u00adecules in the asymmetric unit and intra\u00admolecular P\u2014Rh\u2014P bite angles of 82.78\u2005(5), 82.97\u2005(6) and 83.09\u2005(5)\u00b0 are observed. The dihedral angles between the P\u2014Rh\u2014P and the X\u2014Rh\u2014X planes (X is the centroid of a double bond) are 14.7\u2005(1), 14.8\u2005(1) and 15.3\u2005(1)\u00b0. The structure exhibits disorder of one cyclo\u00adocta\u00addiene ligand as well as one BF4 anion.The title compound, [Rh(C DOI: 10.1107/S1600536810019859/im2198Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "It was synthesized from 2,4,6,8-tetra\u00adphenyl-3,7-diaza\u00adbicyclo\u00ad[3.3.1]nonan-9-one and was crystallized from a methanol\u2013ethanol solution over two years as a racemate. The C=N double bond [1.2868\u2005(15)\u2005\u00c5] is bent significantly out of the plane of the aromatic bicyclic ring system [N\u2014C\u2014C\u2014C = \u2212157.63\u2005(12)\u00b0] and out of the plane of the phenyl ring bonded at the 1-position [N\u2014C\u2014C\u2014C = 41.15\u2005(16)\u00b0].The title compound, C DOI: 10.1107/S1600536810013619/bt5246Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Intra\u00admolecular C\u2014H\u22efO hydrogen bonding and inter\u00admolecular \u03c0\u2013\u03c0 stacking between parallel pyridine rings [centroid\u2013centroid distance = 3.658\u2005(3)\u2005\u00c5] are present in the crystal structure.In the title compound, [CdI DOI: 10.1107/S1600536810012572/xu2734Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The anion is a six-coordinated complex with a distorted octa\u00adhedral geometry around the FeIII atom. Extensive inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, involving the complex anion, (pydaH)+ counter-ion and two uncoordinated water mol\u00adecules, and \u03c0\u2013\u03c0 [centroid-to-centroid distance 3.323\u2005(11)\u2005\u00c5] and C\u2014O\u22ef\u03c0 [O\u2013centroid distance 3.150\u2005(15)\u2005\u00c5] inter\u00adactions connect the various components into a supra\u00admolecular structure.The reaction of iron(II) sulfate hepta\u00adhydrate with the proton-transfer compound (pydaH)(hypydcH) (pyda = pyridine-2,6-diamine; hypydcH DOI: 10.1107/S1600536808029280/hy2152Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The pyrrolidine ring is almost perpendicular to the cyclo\u00adpenta\u00adnone ring, making a dihedral angle of 81.91\u2005(6)\u00b0. The mol\u00adecular conformation is stabilized by an intra\u00admolecular O\u2014H\u22efN hydrogen bond and C\u2014H\u22efO inter\u00adactions. The crystal structure is stabilized by inter\u00admolecular C\u2014H\u22efO inter\u00adactions.In the title compound, [Fe(C DOI: 10.1107/S1600536809018583/bt2949Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CuII center has a distorted square-pyramidal coordination with three N atoms of the 4\u2032-ferrocenyl-2,2\u2032:6\u2032,2\u2032\u2032- terpyridine (fctpy) ligand and one 1,10-phenanthroline (phen) N atom in the basal plane and a second phen N atom in the apical position with an axial distance of 2.254\u2005(4)\u2005\u00c5. The disordered ClO4 \u2212 anion is weakly coordin\u00adated to the CuII ion with a Cu\u2014O distance of 2.766\u2005(11)\u2005\u00c5. The two cyclo\u00adpenta\u00addienyl rings of the ferrocenyl group are almost eclipsed with a deviation of 4.7\u2005(1) \u00b0, and are involved in inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions with the outer pyridyl rings of the fctpy ligands [centroid\u2013centroid distance = 3.759\u2005(2)\u2005\u00c5.].The title complex, [CuFe(C DOI: 10.1107/S1600536809020753/at2803Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The bis\u00ad(diphenyl\u00adarsino)methane ligand bridges an Ru\u2014Ru bond and the monodentate phosphine ligand bonds to the third Ru atom. Both the phosphine and arsine ligands are equatorial with respect to the Ru3 triangle. In addition, each Ru atom carries one equatorial and two axial terminal carbonyl ligands. With regard to the three phosphine-substituted rings, the benzyl ring makes dihedral angles of 41.0\u2005(3) and 43.9\u2005(3)\u00b0 with the other two benzene rings in mol\u00adecule A; these angles are 49.8\u2005(3) and 56.8\u2005(3)\u00b0 in mol\u00adecule B. The dihedral angles between the two benzene rings are 76.1\u2005(3) and 88.2\u2005(3)\u00b0 for the two diphenyl\u00adarsino groups in mol\u00adecule A and 71.3\u2005(3) and 78.1\u2005(3)\u00b0 in mol\u00adecule B. In the crystal packing, mol\u00adecules are linked into chains via inter\u00admolecular C\u2014H\u22efO hydrogen bonds. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions further stabilize the crystal structure.The asymmetric unit of the title DOI: 10.1107/S1600536809053045/sj2709Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The average Ni\u2014N and Ni\u2014O bond lengths are 2.131\u2005(13) and 2.098\u2005(11)\u2005\u00c5, respectively. An intramolecular N\u2014H\u22efO inter\u00adaction occurs. Relatively weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions between the ligands and the ClO4 \u2212 ions result in a chain extending along the b axis.In the title complex, [Ni(CH DOI: 10.1107/S1600536810034446/pv2321Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The Co\u2014N bonds involving the thio\u00adcyanate ligands are significantly shorter than the other two Co\u2014N bonds. The atoms of one of the eth\u00adoxy groups are essentially coplanar with the phenanthroline ring [N=C\u2014O\u2014C = 178.8\u2005(4)\u00b0], while the other eth\u00adoxy group is slightly twisted from the phenanthroline ring plane [N=C\u2014O\u2014C = 167.2\u2005(4)\u00b0]. In the crystal structure, there is a weak \u03c0\u2013\u03c0 stacking inter\u00adaction between two symmetry-related phenanthroline rings with a centroid\u2013centroid distance of 3.706\u2005(4)\u2005\u00c5.In the title complex, [Co(NCS) DOI: 10.1107/S160053680803496X/lh2718Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The coordination bonds of the 4,4\u2032-substituted bipyridyl donors [Ru\u2014N = 2.038\u2005(3) and 2.051\u2005(3)\u2005\u00c5] are shorter than those of the 2,2\u2032-bipyridyl donors [Ru\u2014N1 = 2.065\u2005(3)\u20132.077\u2005(3)\u2005\u00c5], due to the electron-withdrawing effects of the substituents at the 4,4\u2032-positions. The angles between the pyridyl planes of the three bipyridyl ligands are 1.5\u2005(2), 6.3\u2005(3) and 8.7\u2005(2)\u00b0, respectively. The cations are connected by anions via N\u2014H\u22efF inter\u00adactions.In the title compound, [Ru(C DOI: 10.1107/S1600536809002360/kp2198Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The [ZnCl4]2\u2212 anions have a distorted tetra\u00adhedral geometry. The dihedral angles between the isoquinoline rings of the two cations are nearly equal [16.1\u2005(2) and 16.3\u2005(2)\u00b0]. In the crystal structure, the ordered linear formation is aggregated by weak inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions between neighboring isoquinoline pyridine rings with a centroid\u2013centroid distance of 3.779\u2005(4)\u2005\u00c5.The asymmetric unit of the title compound, (C DOI: 10.1107/S1600536809017693/bq2137Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The geometry of the resulting CuN2O4 coordination can be described as distorted octa\u00adhedral. The the two pydc2\u2212 fragments are almost perpendicular to one another [77.51\u2005(11)\u00b0]. To balance the charges, two centrosymmetric protonated butane-1,4-diammonium, (bdaH2)2+ cations are present. In the crystal structure, extensive O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds [D\u22efA = 2.720\u2005(2)\u20133.446\u2005(3)\u2005\u00c5], ion pairing, C\u2014O\u22ef\u03c0 [O\u22ef\u03c0 = 3.099\u2005(2)\u2005\u00c5] and \u03c0\u2013\u03c0 stacking inter\u00adactions between the pydc2\u2212 rings [centroid\u2013centroid distance = 3.5334\u2005(15)\u2005\u00c5] contribute to the formation of a three-dimensional supra\u00admolecular structure.In the title compound, (C DOI: 10.1107/S1600536808011938/su2053Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, N\u2014H\u22efO hydrogen bonds link mol\u00adecules in rows along a. Short inter\u00admolecular Cl\u22efCl inter\u00adactions [3.4225\u2005(5)\u2005\u00c5] link these rows into sheets in the ac plane. Additional weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions generate a three-dimensional network.In the title benzamide derivative, C DOI: 10.1107/S1600536808036313/sg2272Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The metal centre has a distorted octa\u00adhedral coordination with the monoanionic Schiff base ligand occupying one equatorial and two axial coordination positions. The separation between V atoms is 3.214\u2005(3)\u2005\u00c5. In the crystal structure, there are N\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22ef\u03c0 hydrogen bonds, and \u03c0\u2013\u03c0 inter\u00adactions.In the title dinuclear compound, [V DOI: 10.1107/S160053680801787X/xu2429Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by aromatic \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of neighbouring mol\u00adecules [centroid-to-centroid distance = 3.635\u2005(3)\u2005\u00c5], and by C\u2014H\u22ef\u03c0 inter\u00adactions between a propyl methyl\u00adene H atom and the furan ring of an adjacent mol\u00adecule. In addition, the crystal structure exhibits weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds.In the title compound, C DOI: 10.1107/S1600536809004735/gk2190Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Inter\u00adactions of the \u03c0\u2013\u03c0 type are absent between cations in the stacks [centroid\u2013centroid separation = 5.01\u2005(5)\u2005\u00c5]. Significant inter\u00admolecular Br\u2013aryl inter\u00adactions are present in the structure, especially an unusually short Br\u2013ring centroid inter\u00adaction of 3.78\u2005(1)\u2005\u00c5. The coordination geometry of the anion is approximately tetrahedral and a twofold rotation axis passes through the Co atom.In the crystal structure of the title compound, (C DOI: 10.1107/S160053680800439X/at2543Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The 2-fluoro\u00adbenzoate ligands bridge Pb atoms, giving rise to a one-dimensional chain structure extending along the [100] direction. The polymeric chains are connected via C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions, with an inter\u00adplanar distance of 3.46\u2005(1)\u2005\u00c5. An intramolecular O\u2014H\u22efF interaction is also present.In the title compound, [Pb DOI: 10.1107/S1600536808022678/hy2143Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Face-to-face \u03c0-stacking inter\u00adactions between inversion-related pyridine rings with centroid\u2013centroid distances of 3.548\u2005(3) and 3.662\u2005(3)\u2005\u00c5 (perpendicular distances between the respective rings are 3.314 and 3.438\u2005\u00c5) are found. Inter\u00admolecular O\u2014H\u22efO hydrogen bonds between water mol\u00adecules and L ligands form R 5 3(10), R 6 5(14) and R 5 5(12) rings and also a centrosymmetric cage-like unit of water mol\u00adecules, which link eight adjacent NiII centers, forming a three-dimensional framework.In the title compound, [Ni(C DOI: 10.1107/S1600536809006515/si2152Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II atom of the title complex, [Cu(C17H15N2S2)2], lies on a twofold rotation axis, and is in a distorted tetra\u00adhedral geometry with the two bidentate N2S2 Schiff bases. In the crystal structure, the mol\u00adecules are inter\u00adconnected into chains along the c axis by weak C\u2014H\u22efS inter\u00admolecular inter\u00adactions. The crystal packing is further stabilized by C\u2014H\u22ef\u03c0 inter\u00adactions.The Cu DOI: 10.1107/S1600536808002262/ci2558Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The dihedral between the naphthalene ring system and the bridging carbonyl C\u2014C(=O)\u2014C plane is 54.68\u2005(6)\u00b0, far larger than that [12.54\u2005(7)\u00b0] between the phenyl group and the bridging carbonyl group. The nitro group and the phenyl ring are almost coplanar [O\u2014N\u2014C\u2014C torsion angle = 2.94\u2005(19)\u00b0]. In the crystal, mol\u00adecules are linked by C\u2014H\u22ef\u03c0 inter\u00adactions and the phenyl rings are involved in a centrosymmetric \u03c0\u2013\u03c0 inter\u00adaction with a perpendicular distance of 3.523\u2005\u00c5 and a lateral offset of 1.497\u2005\u00c5. In addition, weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds are formed between an H atom of one meth\u00adoxy group and a nearby carbonyl O atom.In the title compound, C DOI: 10.1107/S1600536810005398/fl2290Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The observed Pt\u2014I bond distances of 2.6094\u2005(5) and 2.6130\u2005(5)\u2005\u00c5 are in the expected range for PtI2 complexes. The C=C double bonds in the mol\u00adecule differ significantly [1.373\u2005(10) and 1.403\u2005(10)\u2005\u00c5]. As expected for a platinum(II) complex, the PtII atom is in a square-planar environment (\u03a3Pt\u03b1= 359.71\u00b0).The monoclinic title complex, [PtI DOI: 10.1107/S1600536809029997/bg2283Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The AgI atom is coordinated in an almost linear fashion by two pyridyl N atoms from two nicotinate ligands. The linear coordination is augmented by weak inter\u00adactions with three O atoms from one perchlorate anion, one uncoordinated water mol\u00adecule and one carboxyl\u00adate group. Two Pr atoms link two {Ag(nic)2}+ units into a ring, which is further extended into an infinite zigzag chain by sharing the Pr atoms. These chains are further connected into a three-dimensional network via weak Ag\u22efO inter\u00adactions, O\u2014H\u22efO hydrogen bonds, Ag\u22efAg inter\u00adactions [3.357\u2005(2)\u2005\u00c5] and \u03c0\u2013\u03c0 inter\u00adactions between the pyridyl rings [centroid\u2013centroid distance = 3.685\u2005(4)\u2005\u00c5].In the title compound, {[Ag DOI: 10.1107/S1600536809001718/hy2177Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The bis\u00ad(diphenyl\u00adarsino)methane ligand bridges an Ru\u2014Ru bond and the monodentate phosphine ligand bonds to the third Ru atom. Both the phosphine and arsine ligands are equatorial with respect to the Ru3 triangle. In addition, each Ru atom carries one equatorial and two axial terminal carbonyl ligands. The three phosphine-substituted benzene rings make dihedral angles of 73.5\u2005(3), 57.2\u2005(3) and 75.7\u2005(3)\u00b0 with each other in mol\u00adecule A, while these angles are 60.7\u2005(3), 86.8\u2005(3) and 54.9\u2005(3)\u00b0 in mol\u00adecule B. The dihedral angles between the two benzene rings are 87.3\u2005(3) and 89.6\u2005(3)\u00b0 for the two diphenyl\u00adarsino groups in mol\u00adecule A and 85.6\u2005(3) and 87.7\u2005(3)\u00b0 in mol\u00adecule B. In the crystal packing, the mol\u00adecules are linked into a three-dimensional framework via inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions furture stabilize the crystal structure. The crystal studied was an inversion twin, the refined ratio of twin components being 0.480\u2005(7):0.520\u2005(7).The asymmetric unit of the title DOI: 10.1107/S1600536809053884/sj2702Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The benzofuran ring plane makes dihedral angles of 28.63\u2005(6) and 31.55\u2005(5)\u00b0 with the 4-fluoro\u00adphenyl and phenyl rings, respectively. Weak C\u2014H\u22efF and C\u2014H\u22efO hydrogen bonds and inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions are present in the crystal structure. The title crystal was refined as an inversion twin with a 0.39\u2005(7):0.61\u2005(7) domain ratio.In the title mol\u00adecule, C DOI: 10.1107/S1600536810014613/fb2185Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit contains two PbII atoms, four MBA ligands and two water mol\u00adecules. Each PbII cation is hepta\u00adcoordinated and chelated by four carboxyl\u00adate O atoms from two MBA ligands. The Pb atoms are bridged through the carboxyl\u00adate O atoms from another two MBA ligands, leading to a central Pb2O2 core. The Pb\u2014O bond lengths are in the range 2.325\u2005(3)\u20132.757\u2005(4)\u2005\u00c5. The intra- and inter\u00addimer Pb\u22efPb distances are 4.2942\u2005(3) and 4.2283\u2005(3)\u2005\u00c5, respectively, indicating little direct metal\u2013metal inter\u00adaction. The coordinating water mol\u00adecules and carboxyl\u00adate O atoms are involved in extensive O\u2014H\u22efO hydrogen-bonding inter\u00adactions. The complex has an extended ladder-like chain structure and the chains are assembled by hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distance = 3.6246\u2005(3)\u2005\u00c5] into a three-dimensional supra\u00admolecular structure.The reaction of lead(II) acetate and 3-methyl\u00adbenzoic acid (MBA) in aqueous solution yielded the title polymer, [Pb(C DOI: 10.1107/S1600536809019771/fj2218Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "H)-thiolate anions in the asymmetric unit of the title compound, Na+\u00b7C2H2N3S2 \u2212\u00b72H2O, which are almost perpendicular to each other [dihedral angle = 84.64\u2005(6)\u00b0]. The two Na+ cations are in distorted fourfold coordinations by O atoms of the water molecules. The crystal structure is stabilized by N\u2014H\u22efS, O\u2014H\u22efN and O\u2014H\u22efS hydrogen bonds.There are two 5-amino-1,3,4-thia\u00addiazole-2(3 DOI: 10.1107/S1600536809051897/pk2208Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In addition to the usual inter\u00admolecular C\u2014H\u22efO hydrogen bonding, short intra\u00admolecular C\u2014H\u22efS contacts and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid distance = 3.762\u2005(2)\u2005\u00c5] contribute to the crystal packing.In the title compound, C DOI: 10.1107/S1600536808007356/rk2081Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the crystal structure, inter\u00admolecular N\u2014H\u22efBr hydrogen bonds link the mol\u00adecules into centrosymmetric dimers. There are \u03c0\u2013\u03c0 contacts between the pyridine rings [centroid\u2013centroid distances = 3.9662\u2005(5) and 3.9321\u2005(4)\u2005\u00c5]. There also exists a C\u2014H\u22ef\u03c0 contact between the pyridine CH group and a pyridine ring.In the mol\u00adecule of the title compound, [HgBr DOI: 10.1107/S1600536808038129/hk2575Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "An inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adaction between the pyridine rings [centroid\u2013centroid distance = 3.900\u2005(4)\u2005\u00c5] is observed. The CdII atom has a distorted tetra\u00adhedral coordination.In the cation of the title compound, (C DOI: 10.1107/S1600536808030092/is2330Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The centroid\u2013centroid distances of 3.809\u2005(2) and 3.680\u2005(2)\u2005\u00c5 between nearly parallel pyridine rings of the phen ligands and the benzene rings of dhba anions indicate that the dhba anions are involved in \u03c0\u2013\u03c0 stacking in the crystal structure. The face-to-face separation of 3.35\u2005(3)\u2005\u00c5 between parallel phen ring systems also suggests \u03c0\u2013\u03c0 stacking between adjacent complex mol\u00adecules. The crystal structure contains extensive O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonding.In the crystal structure of the title compound, [Cd(C DOI: 10.1107/S1600536808018126/rk2092Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by various \u03c0\u2013\u03c0 stacking inter\u00adactions in which the benzene ring, a pyridine ring and the five-membered CuN2C2 ring are involved. The centroid\u2013centroid distances range from 3.5631\u2005(15) to 3.5666\u2005(16)\u2005\u00c5.In the title mononuclear complex, [CuCl DOI: 10.1107/S160053680801948X/lh2647Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is composed of centrosymmetric dimers lying about inversion centres. Both independent Sn atoms adopt distorted trigonal-bipyramidal SnC2O3 coordination geometries with the basal planes consisting of two C-atoms from the methyl groups and a bridging O atom. The Sn\u2014C and Sn\u2014O bond lengths lie in the ranges 2.090\u2005(2)\u20132.104\u2005(3) and 2.0241\u2005(14)\u20132.2561\u2005(15)\u2005\u00c5, respectively. The central four-membered planar Sn2O2 ring [Sn\u22efSn distance = 3.2993\u2005(2)\u2005\u00c5] makes dihedral angles of 5.43\u2005(11) and 59.50\u2005(7)\u00b0 with the methyl\u00adphenyl groups, which are themselves oriented at a dihedral angle of 61.38\u2005(8)\u00b0. Besides weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions, the packing mainly features van der Waals forces between the mol\u00adecules.The title compound, [Sn DOI: 10.1107/S1600536810036512/wm2399Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit contains one 4-methyl\u00adbenzoate anion, one nicotinamide (NA) ligand and one coordinated water mol\u00adecule. The four O atoms in the equatorial plane around the MnII ion form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two pyridine N atoms of the NA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the attached benzene ring is 9.01\u2005(7)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 42.44\u2005(5)\u00b0. In the crystal structure, inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, and O\u2014H\u22ef\u03c0 and C\u2014H\u22ef\u03c0 inter\u00adactions link the mol\u00adecules into a two-dimensional network parallel to (001).In the mononuclear title complex, [Mn(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C6H6N2O)2(H2O)2] = 0.035 wR(F 2) = 0.099 S = 1.08 3297 reflections204 parameters3 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.73 e \u00c5\u22123 \u0394\u03c1min = \u22120.38 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008ORTEP-3 for Windows (Farrugia, 1997WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S1600536810011815/ci5069sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810011815/ci5069Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "In the absence of any strong hydrogen bonds, the structure results from a large number of competing weaker inter\u00adactions including Cl\u22efCl [3.4610\u2005(5)\u2005\u00c5] and C\u2014H\u22efCl contacts and both (aryl) C\u2014H\u22efBr and N+\u2014Csp 3\u2014H\u22efBr\u2212 cation\u2013anion inter\u00adactions.In the title compound, C DOI: 10.1107/S160053680902159X/jh2078Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The 2,6-dihydroxy\u00adbenzoic acid mol\u00adecule is disordered about an inversion center. The face-to-face separations of 3.540\u2005(11) and 3.429\u2005(8)\u2005\u00c5 between parallel phen ligands indicate the existence of \u03c0\u2013\u03c0 stacking between adjacent MnII complexes. Uncoordinated water mol\u00adecules are linked with complex and dihydroxy\u00adbenzoic acid mol\u00adecules via O\u2014H\u22efCl and O\u2014H\u22efO hydrogen bonds.In the crystal structure of the title compound, [MnCl DOI: 10.1107/S1600536808012427/ng2452Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The crystal structure is stabilized by N\u2014H\u22efO, N\u2014H\u22efN, O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds. In addition, weak \u03c0\u2013\u03c0 inter\u00adactions are observed between symmetry-related thia\u00addiazole ring systems .The title compound, C DOI: 10.1107/S1600536809021825/at2800Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The AgI atom is coordinated in an almost linear fashion by two pyridyl N atoms of two nic ligands. The linear coordination is augmented by weak inter\u00adactions with one O atom from a half-occupied ClO4 \u2212 anion and a water mol\u00adecule lying on a twofold axis. Two Ag(nic)2 units connect two La atoms, forming a cyclic unit. These units are further extended into an infinite zigzag chain. The chains are bridged by the disordered perchlorate ions via weak Ag\u2014O [2.678\u2005(2)\u2005\u00c5] inter\u00adactions. O\u2014H\u22efO hydrogen bonds, weak Ag\u22efAg [3.3340\u2005(15)\u2005\u00c5] inter\u00adactions and \u03c0\u2013\u03c0 inter\u00adactions between the pyridyl rings [centroid\u2013centroid distance = 3.656\u2005(2)\u2005\u00c5] lead to a three-dimensional network.In the title complex, [Ag DOI: 10.1107/S1600536809026130/hy2200Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The mol\u00adecule has a short intra\u00admolecular contact involving an aromatic H atom (Au\u22efH = 2.64\u2005\u00c5); two adjacent mol\u00adecules are linked by an Au\u22efHylid inter\u00adaction (Au\u22efH = 3.14\u2005\u00c5).The metal atom in the title ylid\u2013gold(I) adduct, [Au(C DOI: 10.1107/S160053680904152X/ng2660Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The geometry of the resulting NiN2O4 coordination can be described as distorted octa\u00adhedral. Considerable C=O\u22ef\u03c0 stacking inter\u00adactions are observed between the carboxyl\u00adate C=O groups and the pyridine rings of the (pydc)2\u2212 fragments, with O\u22ef\u03c0 distances of 3.1563\u2005(12) and 3.2523\u2005(12)\u2005\u00c5 and C=O\u22ef\u03c0 angles of 95.14\u2005(8) and 94.64\u2005(8)\u00b0. In the crystal structure, a wide range of non-covalent inter\u00adactions, consisting of hydrogen bonding , ion pairing, \u03c0\u2013\u03c0 [centroid-to-centroid distance = 3.4825\u2005(8)\u2005\u00c5] and C=O\u22ef\u03c0 stacking, connect the various components to form a supra\u00admolecular structure.The reaction of nickel(II) nitrate hexa\u00adhydrate, propane-1,2-diamine and pyridine-2,6-dicarboxylic acid in a 1:2:2 molar ratio in aqueous solution resulted in the formation of the title compound, (C DOI: 10.1107/S1600536808016309/su2054Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The CdII ion is hexa\u00adcoordinated by four carboxylate O atoms [Cd\u22efO = 2.280\u2005(2)\u20132.404\u2005(2)\u2005\u00c5] from two chelating 4-methoxy\u00adbenzoate anions, and two N atoms [Cd\u22efN = 2.313\u2005(2) and 2.332\u2005(2)\u2005\u00c5] from one chelating 2,2\u2032-dimethyl-3,3\u2032-(oxydiethyl\u00adene)bis\u00ad(1H-benzimidazole) ligand. In the crystal, mol\u00adecules are linked by a weak inter\u00admolecular C\u2014H\u22efO hydrogen bond and an inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction.The title complex, [Cd DOI: 10.1107/S1600536810009505/lx2137Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The structure is consolidated by inter\u00admolecular O\u2014H\u22efO hydrogen bonding, as well as by \u03c0\u2013\u03c0 stacking inter\u00adactions between adjacent naphthyl ring systems [centroid\u2013centroid distance between parallel naphthoate rings is 3.768\u2005(2)\u2005\u00c5].In the title mononuclear complex, [Mg(C DOI: 10.1107/S1600536808006351/ng2419Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "II\u2013HoIII complex ]diphenolato-1\u03ba4 O 1,O 1\u2032,O 6,O 6\u2032:2\u03ba4 O 1,N,N\u2032,O 1\u2032}trinitrato-1\u03ba6 O,O\u2032-holmium(III)nickel(II)), [HoNi(C20H22N2O4)(NO3)3], with the hexa\u00addentate Schiff base compartmental ligand N,N\u2032-bis\u00adethyl\u00adenediamine (H2 L), the Ho and Ni atoms are doubly bridged by two phenolate O atoms of the Schiff base ligand. The coordination of Ni is square-planar with the donor centers of two imine N atoms and two phenolate O atoms. The holmium(III) center has a tenfold \u00adcoordination environment of O atoms, involving the phenolate O atoms, two eth\u00adoxy O atoms and two O atoms each from the three nitrates. Weak C\u2014H\u22efO and O\u22efNi [3.383\u2005(4)\u2005\u00c5] inter\u00adactions generate a two-dimensional zigzag sheet.In the title heteronuclear Ni DOI: 10.1107/S1600536808013755/hg2401Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The asymmetric unit contains one 4-methyl\u00adbenzoate (PMB) anion, one N,N-diethyl\u00adnicotinamide (DENA) ligand and one coordinated water mol\u00adecule. The four O atoms in the equatorial plane around the CoII ion form a slightly distorted square-planar arrangement, while the slightly distorted octa\u00adhedral coordination is completed by the two pyridine N atoms of the DENA ligands in the axial positions. The dihedral angle between the carboxyl\u00adate group and the attached benzene ring is 3.73\u2005(14)\u00b0, while the pyridine and benzene rings are oriented at a dihedral angle of 77.28\u2005(6)\u00b0. In the crystal structure, inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a two-dimensional network parallel to (001). The structure is further stabilized by \u03c0\u2013\u03c0 contacts between the pyridine rings [centroid\u2013centroid distance = 3.544\u2005(1)\u2005\u00c5] and weak C\u2014H\u22ef\u03c0 inter\u00adactions involving the benzene ring.In the centrosymmetric mononuclear title complex, [Co(C N,N-diethyl\u00adnicotinamide, see: Bigoli et al. 2(C10H14N2O)2(H2O)2] = 0.038 wR(F 2) = 0.090 S = 1.04 4484 reflections234 parameters3 restraintsH atoms treated by a mixture of independent and constrained refinementmax = 0.86 e \u00c5\u22123 \u0394\u03c1min = \u22120.55 e \u00c5\u22123 \u0394\u03c1 APEX2 used to solve structure: SHELXS97 (Sheldrick, 2008SHELXL97 (Sheldrick, 2008et al., 2006WinGX (Farrugia, 1999PLATON (Spek, 2009Data collection: 10.1107/S1600536810013954/ci5076sup1.cif Crystal structure: contains datablocks I, global. DOI: 10.1107/S1600536810013954/ci5076Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "There are three N-[(anthracen-9-yl)\u00admethyl\u00adene\u00adamino]\u00adthio\u00adureate ligands coordinated to the CoIII atom via three imine N and three thio\u00adamide S atoms. The Co\u2014S and Co\u2014N bond distances are in expected ranges . The endocyclic S\u2014Co\u2014N bond angles in the five-membered chelate rings range from 82.91\u2005(7) to 85.33\u2005(7)\u00b0. The structure contains four water mol\u00adecules which are disordered over 12 sites and link the complex mol\u00adecules into a three-dimensional network through N\u2014H\u22efO, O\u2014H\u22efO, O\u2014H\u22efN, and O\u2014H\u22efS hydrogen bonds.In the title complex, [Co(C DOI: 10.1107/S1600536808031425/pv2097Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "The two inversion-related CdII ions are separated by 3.9920\u2005(6)\u2005\u00c5 and are bridged by two O atoms from two nitrite ligands. There are two types of \u03c0\u2013\u03c0 stacking inter\u00adactions involving symmetry-related pyrazole rings, with centroid\u2013centroid distances of 3.445\u2005(2) and 3.431\u2005(2)\u2005\u00c5.In the title centrosymmetric binuclear complex, [Cd DOI: 10.1107/S1600536809039841/lh2917Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "AID). Upon germinal centre exit, B cells differentiate into antibody\u2010secreting plasma cells. Germinal centre maintenance and terminal fate choice require transcriptional reprogramming that associates with a substantial reconfiguration of DNA methylation patterns. Here we examine the role of ten\u2010eleven\u2010translocation (TET) proteins, enzymes that facilitate DNA demethylation and promote a permissive chromatin state by oxidizing 5\u2010methylcytosine, in antibody\u2010mediated immunity. Using a conditional gene ablation strategy, we show that TET2 and TET3 guide the transition of germinal centre B cells to antibody\u2010secreting plasma cells. Optimal AID expression requires TET function, and TET2 and TET3 double\u2010deficient germinal centre B cells show defects in CSR. However, TET2/TET3 double\u2010deficiency does not prevent the generation and selection of high\u2010affinity germinal centre B cells. Rather, combined TET2 and TET3 loss\u2010of\u2010function in germinal centre B cells favours C\u2010to\u2010T and G\u2010to\u2010A transition mutagenesis, a finding that may be of significance for understanding the aetiology of B\u2010cell lymphomas evolving in conditions of reduced TET function.Upon activation by antigen, B cells form germinal centres where they clonally expand and introduce affinity\u2010enhancing mutations into their B\u2010cell receptor genes. Somatic mutagenesis and class switch recombination (CSR) in germinal centre B cells are initiated by the activation\u2010induced cytidine deaminase . Loss of TET function favors C\u2010to\u2010T and G\u2010to\u2010A mutagenesis during somatic hypermutation, a finding of potential significance for understanding the etiology of B\u2010cell lymphomas.TET (ten\u2010eleven\u2010translocation) enzymes promote gene expression by catalyzing the oxidation of 5\u2010methylcytosine in DNA. TET deficiency has been linked to defects in embryonic development and enhanced risk of myeloid malignancies, including B\u2010cell lymphomas. Here, Verena Labi and colleagues show using a conditional gene ablation strategy that TET function is vital for humoral immunity As compared to mature na\u00efve follicular (FO) B cells, TET2 and TET3 are substantially down\u2010regulated in antigen\u2010experienced GC B cells and plasma cells, a result in agreement with a recent report in human GC B cells During B\u2010cell development expression of both, TET2 Fig.\u00a0A and TETDependent on the mouse strain and animal facility, young mice lacking only TET2 or TET3 display no or only moderate B\u2010cell phenotypes at steady state in\u00a0vitro treatment of activated B cells with 5\u2010azacytidine augmented the appearance of plasmablasts in a division\u2010dependent manner B\u2010cell transit through the GC is accompanied by extensive DNA demethylation, focal methylation gains and an overall increased heterogeneity in DNA methylation patterns ;Tet2F/F;Tet3F/F mice in which physiologic germ\u2010line Cg1 transcription drives expression of the Cre\u2010recombinase Tet genes is expected in a majority of GC B cells upon IgG1\u2010priming. Of note, acute GC B cell\u2010specific Tet deletion circumvents indirect effects caused by extended TET\u2010deficiency during B\u2010cell development.Addressing the involvement of TET proteins in this process, we generated Cg1\u2010Crein\u00a0vitro induced GC (iGC) B cells First, we used a co\u2010culture system that allows the generation and exponential growth of Tet deletion is complete as indicated by qRT\u2010PCR analysis conjugated to chicken gamma globulin (NP\u2010CGG). NP\u2010specific GC B cells that form in response to alum\u2010adjuvanted immunogens predominately switch to IgG1 ;Tet2F/F;Tet3F/F GC B cells, suggesting that these cells proliferate normally and high\u2010affinity (NP1.7\u2010BSA) \u03b1\u2010NP IgG1 were decreased similarly, resulting in no significant skewing of the affinity ratio in Cg1\u2010Cre;Tet2F/F;Tet3F/F mice ;Tet2F/F;Tet3F/F (54%) GC B\u2010cell clones in PBS in a volume of 200\u00a0\u03bcL per mouse. Animal procedures were approved by the Austrian Federal Ministry of Education, Science and Research (BMWF: 66\u2010011/0106\u2010WF/3b/2015 and 66\u2010011/0031\u2010V/3b/2019).The Cg1\u2010Cre 4)2 in PBS] was mixed with a similar volume of 1\u00a0mg\u00b7mL\u22121 NP15\u2010CGG , and the pH was adjusted to 6.5\u20137.0 using 10\u00a0m NaOH . The precipitate was washed three times with PBS at 2348 g\u00a0for 15\u00a0s and finally resuspended in PBS to reach the initial volume of the Alum/NP\u2010CGG mixture.A freshly prepared 10% Alum solution was generally used for cell preparation. Single\u2010cell suspensions were prepared by forcing murine spleen through 70\u00a0\u03bcB cells were enriched from splenic single\u2010cell suspensions using MagniSort Streptavidin Negative Selection Beads as per manufacturer's instructions, and biotinylated antibodies against Ter119 , CD11b and TCR\u03b2 .et\u00a0al. m l\u2010glutamine and 100\u00a0U\u00b7mL\u22121\u00a0penicillin/100\u00a0\u03bcg\u00b7mL\u22121\u00a0streptomycin . 3\u00a0\u00d7\u00a0106 feeder cells per 10\u00a0cm plate were treated for 2\u00a0h with 10\u00a0\u03bcg\u00b7mL\u22121 mitomycin C in 6\u00a0mL feeder cell medium. Following five repeated washing steps with PBS, 1.5\u00a0\u00d7\u00a0106 B cells/10\u00a0cm dish were plated on 40LB feeder cells in 40\u00a0mL B\u2010cell medium: DMEM supplemented with 10% (v/v) FBS, 2\u00a0mm l\u2010glutamine, 10\u00a0mm Hepes , 1\u00a0mm sodium pyruvate , 1\u00d7 nonessential amino acids , 100\u00a0U\u00b7mL\u22121\u00a0penicillin/100\u00a0\u03bcg\u00b7mL\u22121\u00a0streptomycin, 50\u00a0\u03bcm \u03b2\u2010mercapto\u2010ethanol and 10\u00a0ng\u00b7mL\u22121 rIL\u20104 . On day 3, 30\u00a0mL of medium containing IL\u20104 were replaced. On day 4, iGC B cells were harvested and analysed. 1.5\u00a0\u00d7\u00a0106 iGC B cells were replated per 10\u00a0cm dish containing fresh mitomycin C\u2010treated 40LB feeder cells as detailed above, and cultivated in 40\u00a0mL of B\u2010cell medium containing either 10\u00a0ng\u00b7mL\u22121 rIL\u20104 or 10\u00a0ng\u00b7mL\u22121 rIL\u201021 . Thirty millilitre of medium containing the respective cytokines were replaced by fresh medium on day 7. The final analysis was conducted on day 8. Analyses (day 4 and 8) entailed collection of medium for ELISA and pellets for qRT\u2010PCR, cell counting (Trypan blue exclusion) and flow cytometric analysis. For analysis, iGC B cells were generally handled at room temperature and cultured at 37\u00a0\u00b0C in a humidified atmosphere containing 5% CO2.The iGC B cell culture was conducted as described by Nojima 2. Mitogenic stimuli were used as follows: 1\u00a0\u03bcg\u00b7mL\u22121 \u03b1\u2010CD40 , 25\u00a0ng\u00b7mL\u22121 IL\u20104 , 10\u00a0ng\u00b7mL\u22121 IL\u201021 (Peprotech 210\u201021), 20\u00a0mg\u00b7mL\u22121 lipopolysaccharide and 25\u00a0ng\u00b7mL\u22121 IL\u20105 .Primary B cells were enriched from splenic single\u2010cell suspensions and cultivated in medium as described above for the iGC B\u2010cell culture at 37\u00a0\u00b0C in a humidified atmosphere containing 5% COin\u00a0vitro, B cells were labelled with 10\u00a0\u03bcm Cell Proliferation Dye eFluor 450 as per manufacturer's instructions and placed in culture.To determine cell proliferation in the presence of mitogens 2\u2010FITC , \u03b1CD25\u2010PE , \u03b1cKit\u2010APC , \u03b1CD1d\u2010PE , \u03b1CD23\u2010PE/Cy7 , \u03b1CD38\u2010FITC , \u03b1CD38\u2010APC , \u03b1CD38\u2010eFluor450 , \u03b1CD95\u2010PE , \u03b1CD95\u2010PE/Cy7 , \u03b1CXCR4\u2010APC , \u03b1CD86\u2010PE/Cy7 , \u03b1IgG1\u2010FITC , \u03b1IgG1\u2010PE , CD138\u2010PE and CD138\u2010BV510 . NP24\u2010PE was used to detect NP\u2010binding B cells. Data were acquired on an LSRII cytometer (BD Biosciences) and analysed using flowjo software . Nonsinglet events were excluded from analyses using FSC\u2010H/FSC\u2010W and SSC\u2010H/SSC\u2010W characteristics.Flow cytometry analysis has been performed as described in Ref. m EDTA). Cells were harvested by addition of B\u2010cell medium and vigorous pipetting.To collect iGC B cells from 10\u00a0cm dishes, 9/10 of the medium was gently removed, and the cells were incubated for 5\u00a0min at 37\u00a0\u00b0C in 3\u00a0mL harvest buffer and surface digested for 4\u00a0min at 37\u00a0\u00b0C. To stop trypsin digestion, 3\u00a0mL of staining buffer (described above for flow cytometry) were added, the cells were pelleted at 1800\u00a0r.p.m. for 4\u00a0min and the supernatant was removed. The cells were transferred to a U\u2010bottom 96 well plate and dead cells were labelled with the Fixable Viability Dye eFluor 780 as per manufacturer's instructions. Subsequently, cells were fixed and processed using the active Caspase 3 staining kit as per manufacturer's instructions. Before staining with the active Caspase 3 antibody, the iGC B cells were incubated with 1\u00a0\u03bcg\u00b7mL\u22121 of \u03b1CD16/32 Fc\u2010Block in 25\u00a0\u03bcL perm/wash buffer for 10\u00a0min at 4\u00a0\u00b0C. Subsequently another 25\u00a0\u03bcL of \u03b1IgE\u2010bio diluted in perm/wash buffer was added, the cells were further incubated for 15\u00a0min at 4\u00a0\u00b0C and washed with perm/wash buffer at 568\u00a0g\u00a0for 2\u00a0min. Next, the cells were incubated with 25\u00a0\u03bcL of antibody mix containing \u03b1IgM\u2010PeCy7 , \u03b1IgG1\u2010FITC , \u03b1CD19\u2010BV605 and Streptavidin\u2010APC for 15\u00a0min at 4\u00a0\u00b0C. The cells were washed with perm/wash buffer at 2000\u00a0r.p.m. for 2\u00a0min and stained for active caspase 3 as per manufacturer's instructions.For CSR analysis, 5\u00a0\u00d7\u00a0105 iGC B cells were transferred to a U\u2010bottom 96 well plate and pelleted at 2000\u00a0r.p.m. for 2\u00a0min. Dead cells were labelled with the Fixable Viability Dye eFluor 780 as per manufacturer's instructions. Subsequently, cells were incubated with 1\u00a0\u03bcg\u00b7mL\u22121 of \u03b1CD16/32 Fc\u2010Block in 25\u00a0\u03bcL staining buffer (PBS/3%FBS) for 10\u00a0min. Next, 25\u00a0\u03bcL of staining buffer containing \u03b1CD19\u2010BV605 and \u03b1CD138\u2010BV421 was added. The cells were further incubated for 15\u00a0min, washed with 200\u00a0\u03bcL of staining buffer at 2000\u00a0r.p.m. for 2\u00a0min, and resuspended in 200\u00a0\u03bcL staining buffer for flow cytometry analysis.The processing of cells for plasmablast flow cytometry analysis was performed at room temperature in the dark. 5\u00a0\u00d7\u00a010flowjo software (Tree Star). Nonsinglet events were excluded from analyses using FSC\u2010H/FSC\u2010W and SSC\u2010H/SSC\u2010W characteristics.Data were acquired on an LSRII cytometer (BD Biosciences) and analysed using \u22121 of \u03b1CD16/32 Fc\u2010Block in 500\u00a0\u03bcL staining buffer (PBS/3%FBS) for 10\u00a0min, washed and stained for 20\u00a0min with antibodies in a volume of 500\u00a0\u03bcL staining buffer. The sorted cell subsets were defined as follows: Bone Marrow: pro B cells (B220loCD19+AA4.1+IgM\u2212CD25\u2212ckit+), large pre B cells (B220loCD19+AA4.1+IgM\u2212CD25+ckit\u2212FSChi), small pre B cells (B220loCD19+AA4.1+IgM\u2212CD25+ckit\u2212FSClo) and immature IgM+ B cells (B220loCD19+AA4.1+IgM+). Spleen: T1 B cells (CD19+B220+AA4.1+CD23\u2212IgMhi), T2 B cells (CD19+B220+AA4.1+CD23+IgMhi), FO B cells (CD19+B220+AA4.1\u2212CD1d+IgM+), MZ B cells (CD19+B220+AA4.1\u2212CD1dhiIgMhi), GC B cells (CD19+B220+CD138\u2212CD95hiCD38lo/\u2212), plasma cells/plasmablasts (B220\u2212/loCD138hi), DZ GC B cells (CD19+B220+CD138\u2212CD95hiCD38lo/\u2212CXCR4hiCD86lo) and LZ GC B cells (CD19+B220+CD138\u2212CD95hiCD38lo/\u2212CXCR4loCD86hi). For sorting B\u2010cell division cycles cells were harvested, filtered through 70\u00a0\u03bcm mesh filters and resuspended in staining buffer.Cell sorting has been performed as described in Ref. Cell sorting was carried out on a FACS Aria III (BD Biosciences). Nonsinglet events were excluded from analyses based on characteristics of FSC\u2010H/FSC\u2010W and SSC\u2010H/SSC\u2010W.in\u00a0vitro\u2010cultivated B cells was isolated using Trizol reagent as per manufacturer's instructions. DNA was removed using the RQ1 RNAse\u2010free DNase , and RNA was retrieved using GlycoBlue Coprecipitant as per manufacturer's instructions. First\u2010strand cDNA was generated from 100\u00a0ng of total RNA using the iScript cDNA Synthesis Kit and cDNA was amplified using the AceQ qPCR SYBR Green Master Mix as per manufacturer's instructions. The qRT\u2010PCR was run on a StepOnePlus Real\u2010time PCR system and melt curve analysis was performed for every run. The expression of individual mRNAs was normalized to HPRT with the following formula: fold induction\u00a0=\u00a02(\u2212\u0394Ct), where \u0394Ct=Ct(target)\u2212Ct(HPRT). The following primers were used: HPRT F: 5\u2032\u2010GTCATGCCGACCCGCAGTC\u20103\u2032, HPRT R: 5\u2032\u2010AGTCCATGAGGAATAAAC\u20103\u2032; TET2\u00a0F: 5\u2032\u2010GCCAGAAGCAAGAAACCAAG\u20103\u2032, TET2 R: 5\u2032\u2010TTGGAGCAATGACAGTAGCC\u20103\u2032; TET3\u00a0F: 5\u2032\u2010AAGAGTCTGCTGGACACACC\u20103\u2032, TET3 R: 5\u2032\u2010CTCCATGAGTTCCCGGATAG\u20103\u2032; AID F: 5\u2032\u2010GGACTTCGGCCACCTTC\u20103\u2032, AID R: 5\u2032\u2010CATCTCAGAAACTCAGCCACG\u20103\u2032. Ki67\u00a0F: 5\u2032\u2010GAACAGACTTGCTCTGGCCTAC\u20103\u2032, Ki67 R: 5\u2032\u2010 CTTCATAGGCATTCCCTCACTC\u20103\u2032.RNA from snap\u2010frozen cell pellets of FACS\u2010sorted B cells was isolated using the Quick\u2010RNA Micro Prep Kit and DNase digestion as per manufacturer's instructions. RNA from snap\u2010frozen \u22121 capture antibody was coated overnight onto 96\u2010well enzyme\u2010linked immunosorbent assay plates at 4\u00a0\u00b0C . Plates were washed three times with wash buffer (PBS containing 0.05% TWEEN 20), blocked with 100\u00a0\u03bcL per well 1% BSA in PBS for 4\u00a0h at room temperature and washed three more times with wash buffer. Subsequently, wells were incubated over night at 4\u00a0\u00b0C with 100\u00a0\u03bcL per well of mouse serum serially diluted 1\u00a0:\u00a04 in blocking buffer (range 1\u00a0:\u00a0800 to 1\u00a0:\u00a0160\u00a0000). Plates were washed three times with wash buffer and incubated with 100\u00a0\u03bcL per well of HRP\u2010conjugated \u03b1\u2010mouse IgG1 or HRP\u2010conjugated \u03b1\u2010mouse IgM for 4\u00a0h at room temperature. For detection, 100\u00a0\u03bcL of ABTS substrate solution per well was incubated for 20\u00a0min. Absorbance was measured at 405\u00a0nm using a microplate reader .For NP\u2010specific serum IgG1 and IgM titres, ELISA was performed as described in zeiss zen blue 2.5 lite software (Zeiss).IHC was performed on 7\u00a0\u03bcm sections of murine spleens as described in Ref. ;Tet2F/F;Tet3F/F mice. Briefly, IgG1+ GC B cells were FACS\u2010sorted (CD19+B220+CD138\u2212FashiCD38lo/\u2212IgG1+), pelleted and total RNA was isolated using the Quick\u2010RNA Micro Prep Kit including the DNase digestion step as per manufacturer's instructions. First\u2010strand cDNA was generated from 100\u00a0ng of total RNA using the Promega GoScript Reverse transcription system and a gene\u2010specific primer for the IgG1 locus: Cg1 5\u2032\u2010CATGGAGTTAGTTTGGGCAG\u20103\u2032. Subsequently, two rounds of semi\u2010nested PCRs were performed using the Herculase II Fusion DNA polymerase kit as per manufacturer's instructions. Following each round of PCR, the gene product was run on an agarose gel and purified using the peqGold Gel extraction kit as per manufacturer's instructions. For the first PCR, the following primers were used: V186.2 leader 5\u2032\u2010AGCTGTATCATGCTCTTCTTGGCA\u20103\u2032, Cg1 5\u2032\u2010CATGGAGTTAGTTTGGGCAG\u20103\u2032. For the second PCR, the following primers were used: V186.2 nested 5\u2032\u2010CATGCTCTTCTTGGCAGCAACAG\u20103\u2032, Cg1 5\u2032\u2010CATGGAGTTAGTTTGGGCAG\u20103\u2032. For cloning of VH186.2 segments, the pJet1.2 blunt vector was used and cleaved with EcoRV . The purified PCR products were ligated into the vector using T4 ligase as per manufacturer's instructions. Using a standard transformation protocol, DH5\u03b1 bacteria were transformed with the bulk of ligated plasmids, spread onto LB Agar/ampicillin plates and incubated over night at 37\u00a0\u00b0C. For each of the six mice, >10 colonies were transferred into a PCR Master Mix , and the very same colonies were selected on new LB agar plates. For the colony PCR, the following primers were used: pJet1.2\u00a0F 5\u2032\u2010CGACTCACTATAGGGAGAGCGGC\u20103\u2032, pJet1.2 R 5\u2032\u2010AAGAACATCGATTTTCCATGGCAG\u20103\u2032. Cycle conditions were: 1 cycle: 95\u00a0\u00b0C for 1\u00a0min; 30 cycles: 95\u00a0\u00b0C for 15\u00a0s, 56\u00a0\u00b0C for 15\u00a0s, 72\u00a0\u00b0C for 45\u00a0s; 1 cycle: 72\u00a0\u00b0C for 3\u00a0min. The PCR products were loaded onto an agarose gel, and colonies with correct insertions were grown in 2\u00a0mL LB medium over night. Plasmid DNA was extracted using the Monarch Plasmid Miniprep Kit as per manufacturer's instructions. Subsequently, Sanger sequencing was performed using the pJet1.2\u00a0F primer detailed above. Sequences were aligned to the VH186.2 germ line sequence using VBASE2 (http://www.vbase2.org/). Mutation data were only used if the entire sequence was intact, and only unique sequences were included in the analysis. According to these criteria, two sequences from Cg1\u2010Cre;Tet2F/F;Tet3F/F GC B cells had to be excluded.Somatic hypermutation analysis was performed according to Ref. graphpad prism 7 software using Student's t test when comparing two groups, One\u2010way ANOVA and Tukey post hoc test when comparing multiple groups and Mann\u2013Whitney test for SHM analysis. The number of biological repetitions (n) is stated in each figure legend, and every experiment was performed at least twice. Differences between groups were considered statistically significant when P\u00a0<\u00a00.05. In figures, asterisks stand for: *P\u00a0<\u00a00.05; **P\u00a0<\u00a00.01; ***P\u00a0<\u00a00.001; ****P\u00a0<\u00a00.0001.Results are always shown as mean and standard deviation (SD). Graphs were plotted and statistical analysis was performed with The authors declare no conflict of interest.Conceptualization: VL; Methodology: VL; Investigation: KS, SM, SH, ED,\u00a0AA; Resources: VL, SH, ED, AV, KR; Writing \u2013 Original Draft: VL; Visualization: VL; Project Administration: VL; Funding Acquisition: VL."} +{"text": "One Cu atom is coordinated to the 2-eth\u00adoxy\u00adpyrazine mol\u00adecule and two bridging cyanide ligands, equally disordered over two sites. The second Cu atom is coordinated by two disordered over two sites bridging cyanide groups. Two copper\u2013cyanide chains are connected through Cu\u22efCu contact.The title compound, {[Cu(EtOpz)(CN) 4H3OC2H5N2)][Cu(CN)]}n, there are two Cu atoms with different coordination environments. One CuI ion is coordinated in a triangular coordination geometry by the N atom of the 2-eth\u00adoxy\u00adpyrazine mol\u00adecule and by two bridging cyanide ligands, equally disordered over two sites exchanging C and N atoms, thus forming polymeric chains parallel to the c axis. The other Cu atom is connected to two bridging cyanide groups disordered over two sites with an occupancy of 0.5 for each C and N atom, and forming an almost linear polymeric chain parallel to the b axis. In the crystal, the two types of chain, which are orthogonal to each other, are connected by cuprophilic Cu\u22efCu inter\u00adactions [2.7958\u2005(13)\u2005\u00c5], forming two-dimensional metal\u2013organic coordination layers parallel to the bc plane. The coordination framework is further stabilized by weak long-range (electrostatic type) C\u2014H\u22ef\u03c0 inter\u00adactions between cyano groups and 2-eth\u00adoxy\u00adpyrazine rings.In the asymmetric unit of the title coordination compound, {[Cu(CN)(C The Cu1 atom is coordinated to the N atom of a 2-eth\u00adoxy\u00adpyrazine mol\u00adecule [Cu1\u2014N5 = 2.090\u2005(4)\u2005\u00c5]. Two other coordination positions are occupied by bridging cyanide groups, which are equally disordered over two sites, exchanging C and N atoms [Cu1\u2014C1/N1 = 1.905\u2005(4)\u2005\u00c5 and Cu1\u2014C2/N2 = 1.888\u2005(4)\u2005\u00c5], thus forming an irregular triangular coordination geometry where the copper ion is displaced from the centre . The Cu2 atom is coordinated by two cyanide ligands, which are also disordered over two sites with an occupancy of 0.5 for each C and N atom to form an almost linear chain [C4/N4iii\u2014Cu2\u2014C3/N3 = 170.5\u2005(2)\u00b0]. The two CuI centres are connected through a Cu\u22efCu inter\u00adaction [Cu1\u2014Cu2 = 2.7958\u2005(13)\u2005\u00c5] that could be inter\u00adpreted as a cuprophilic contact inter\u00adconnected by Cu\u22efCu contacts and forming two-dimensional layers parallel to (100). The Cu\u22efCu contacts are almost perpendicular to the [Cu2(CN)] chains [C3/N3\u2014Cu2\u2014Cu1 = 89.8\u2005(2)\u00b0 and C4/N4iii\u2014Cu2\u2014Cu1 = 99.7\u2005(2)\u00b0]. At the same time, the Cu2 atom occupies an axial position with respect to the triangular [N(CN)2] coordination environment of Cu1 [C1/N1\u2014Cu1\u2014Cu2 = 70.6\u2005(2)\u00b0 and C2/N2\u2014Cu1\u2014Cu2 = 87.6\u2005(2)\u00b0]. The resulting metal\u2013organic coordination framework is additionally stabilized by weak long-range (electrostatic-type) C\u2014H\u22ef\u03c0 inter\u00adactions between cyanide groups and 2-eth\u00adoxy\u00adpyrazine rings \u2005\u00c5 are also observed .The crystal packing of the title compound Fig.\u00a02 consistset al., 2016catena-[penta\u00adkis\u00ad(\u03bc2-cyano)\u00adtris\u00ad(1-phenyl\u00adpiperazine)penta\u00adcopper] (PhPip) and (CuCN)3(PhPip), associated by Cu\u22efCu pairwise cuprophilic inter\u00adactions, with distances of 2.5586\u2005(10) and 2.6441\u2005(10)\u2005\u00c5. A search of the CSD for two C\u2014N\u2014Cu\u2014C\u2014N fragments with a defined Cu\u22efCu distance less than 2.8\u2005\u00c5 gave 80 hits, among which is an example close to the title structure, i.e.catena-[tetra\u00adkis\u00ad(\u03bc2-cyano)\u00adtetra\u00adcopper(I)] and 2.9644\u2005(6)\u2005\u00c5.A search of the Cambridge Structural Database in 1\u2005ml of H2O, the second layer was a H2O/EtOH mixture and the third layer was a solution of 2-eth\u00adoxy\u00adpyrazine in 0.5\u2005ml of EtOH. After two weeks, colourless block-shaped crystals had formed in the middle layer. The crystals were kept under the mother solution prior to measurement.Crystals of the title compound were obtained by slow diffusion within three layers in a 3\u2005ml glass tube. The first layer was a solution of K[Cu(CN)Uiso(H) = 1.2Ueq(C) for aromatic hydrogens, C\u2014H = 0.97\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for the CH2 group, and C\u2014H = 0.96\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for the CH3 group. A rotating model was used for the methyl group. All cyano ligands are disordered over two sites with occupancies of 0.5. The coordinates of C and N atoms sharing the same sites and their displacement ellipsoids were constrained to be the same.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901901452X/rz5265sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901901452X/rz5265Isup2.hklStructure factors: contains datablock(s) I. DOI: 1961274, 1961274CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "DNA polymerase \u03b8 (Pol\u03b8) is a unique polymerase-helicase fusion protein that promotes microhomology-mediated end-joining (MMEJ) of DNA double-strand breaks (DSBs). How full-length human Pol\u03b8 performs MMEJ at the molecular level remains unknown. Using a biochemical approach, we find that the helicase is essential for Pol\u03b8 MMEJ of long ssDNA overhangs which model resected DSBs. Remarkably, Pol\u03b8 MMEJ of ssDNA overhangs requires polymerase-helicase attachment, but not the disordered central domain, and occurs independently of helicase ATPase activity. Using single-particle microscopy and biophysical methods, we find that polymerase-helicase attachment promotes multimeric gel-like Pol\u03b8 complexes that facilitate DNA accumulation, DNA synapsis, and MMEJ. We further find that the central domain regulates Pol\u03b8 multimerization and governs its DNA substrate requirements for MMEJ. These studies identify unexpected functions for the helicase and central domain and demonstrate the importance of polymerase-helicase tethering in MMEJ and the structural organization of Pol\u03b8. DNA polymerase \u03b8 is a polymerase-helicase essential for microhomology-mediated end-joining (MMEJ) or alternative end-joining of DNA. Here the authors use biochemical and biophysical methods to reveal how full-length human DNA polymerase \u03b8 performs MMEJ at the molecular level. MMEJ functions during S and G2 cell-cycle phases and therefore acts on 3\u2032 single-strand DNA (ssDNA) overhangs generated by 5\u2032\u20133\u2032 exonuclease resection of DSBs, similar to homologous recombination (HR) is a polymerase-helicase fusion protein Fig.\u00a0, that is(HR)Fig.\u00a07,8. The 11. Pol\u03b8-hel is related to HELQ/Hel308 helicases, which are involved in replication and repair13. Recent studies show that Pol\u03b8-hel unwinds short DNA in an ATP-dependent manner with 3\u2032-5\u2032 polarity, similar to HELQ/Hel30812. Pol\u03b8-hel also facilitates annealing of complementary ssDNA in an ATP-independent manner, and anneals RPA coated ssDNA by utilizing the energy of ATP14. Pol\u03b8-hel may also act as an anti-recombinase by counteracting RAD51 activity15. Pol\u03b8-pol is related to bacterial Pol I enzymes such as Klenow fragment, but contains an inactive 3\u2032\u20135\u2032 exonuclease domain16, and exhibits low fidelity DNA synthesis and translesion (TLS) synthesis activities21. The polymerase and helicase include unstructured motifs, and a disordered motif (loop 2) in Pol\u03b8-pol was shown to promote its ssDNA extension and MMEJ activities22.Pol\u03b8 consists of a super-family 2 helicase (Pol\u03b8-hel), a disordered central domain (Pol\u03b8-cen), and an A-family polymerase (Pol\u03b8-pol) Fig.\u00a01,10,11. 7, and that Pol\u03b8 competes with HR15. Wyatt et al. also demonstrated that the efficiency of cellular MMEJ is positively correlated with 3\u2032 ssDNA overhang length8. This report further demonstrated that Ku binds tightly to DNA ends with short 4 nt overhangs, but exhibits very low affinity for 70 nt overhangs8. This reveals a clear mechanistic difference between the respective substrate requirements for non-homologous end-joining (NHEJ) and MMEJ. How Pol\u03b8 functions on long ssDNA overhangs to promote MMEJ, however, remains unknown.The ssDNA overhangs that Pol\u03b8 functions on during MMEJ have become clearer in recent cellular studies. Wyatt et al. demonstrated that relatively long (\u2265\u200945\u201370\u2009nt) ssDNA overhangs promote MMEJ in cells Fig.\u00a0. This is2. Recent biochemical studies similarly show that Pol\u03b8-pol can perform MMEJ on short (\u2009<\u200912\u2009nt) ssDNA23. Although these studies provide insight into the activity of Pol\u03b8-pol, they fail to recapitulate MMEJ on DNA with longer (45\u201370\u2009nt) overhangs that support cellular MMEJ8. In vitro, Pol\u03b8-pol primarily performs snap-back replication on long ssDNA as a result of its intrastrand base-pairing activity22. For example, under physiological buffer conditions with Mg2+, Pol\u03b8-pol efficiently extends relatively long (\u2265\u200926\u2009nt) ssDNA by using the 5\u2032 portion of the ssDNA as a template in cis overhangs in cellular studies8, we hypothesized that the helicase domain within full-length Pol\u03b8 (hereafter referred to as Pol\u03b8) promotes MMEJ by suppressing Pol\u03b8-pol intrastrand pairing. For instance, since Pol\u03b8-hel binds tightly to ssDNA and translocates along ssDNA with 3\u2032\u20135\u2032 directionality13, it may inhibit Pol\u03b8-pol intrastrand pairing by binding and/or translocating along ssDNA upstream from the polymerase in the context of the full-length protein 3\u2032 overhangs are required for MMEJ activity by the isolated polymerase domain (Pol\u03b8-pol)cis Fig.\u00a0. Consistcis Fig.\u00a02,22. Becein Fig.\u00a0.S. cerevisiae using a N-terminal 3xFLAG-tag are equally deficient in MMEJ of long ssDNA ssDNA overhangs to promote MMEJ in cells, we examined Pol\u03b8 MMEJ of long ssDNA (70\u2009nt and 100\u2009nt) which models substrates with long 3\u2032 overhangs Fig.\u00a0. We find23. On longer 26\u2009nt ssDNA with identical microhomology, Pol\u03b8-pol exclusively promotes ssDNAx byproducts overhangs2, demonstrates it performs interstrand pairing without Pol\u03b8-hel. Pol\u03b8-pol, however, fails to perform efficient MMEJ of long ssDNA under various conditions ssDNA which is almost entirely contained within its active site Fig.\u00a0, leftSu and 3E16cts Fig.\u00a0, right, cts Fig.\u00a0-4F. Unexcts Fig.\u00a0, yet is cts Fig.\u00a0. This sucts Fig.\u00a0 and 3E23\u2009nt ssDNA8. Consistent with this, we find that decreasing ssDNA length to 45\u2009nt while maintaining the same 6\u2009bp microhomology significantly reduces Pol\u03b8 MMEJ, yet, the full-length protein still shows significantly higher end-joining activity than Pol\u03b8-pol would be extruded from the enzyme\u2019s DNA exit channel conjugated ssDNA linker in place of the central domain (Pol\u03b8\u0394cen) Fig.\u00a027. Pol\u03b8\u0394ity Fig.\u00a0, and is ity Fig.\u00a0. Remarkaity Fig.\u00a0. Togetheons Fig.\u00a0. Controlons Fig.\u00a0, right, ons Fig.\u00a0, and is pol Fig.\u00a0.2, and the addition of Pol\u03b8-hel in trans fails to stimulate Pol\u03b8-pol MMEJ of this substrate as expected ssDNA compared to Pol\u03b8-pol Fig.\u00a0, we test16. Our previous studies demonstrate that Pol\u03b8-hel exhibits similar activities to the related helicase Hel308, such as ATP-dependent 3\u2032-5\u2032 directional ssDNA translocation activity12. Based on this polarity, Pol\u03b8-hel was modeled facing towards the 5\u2032 terminus. The model superimposes the ssDNA substrate from the Hel308 structure, in which the enzyme translocates along, onto Pol\u03b8-hel28. The primer strand from the Pol\u03b8-pol structure is also included with additional ssDNA linking the two substrates together, resulting in a long contiguous 31 nt ssDNA that is easily bound by the tethered enzymes. In this close configuration along ssDNA Pol\u03b8-pol and Pol\u03b8-hel fit well together and space is readily available for a long flexible linker (e.g. Pol\u03b8-cen). Although this model is speculative, it suggests bivalent ssDNA binding by the fusion protein which would be expected to be in the low nanomolar range.Because covalent attachment of Pol\u03b8-hel to Pol\u03b8-pol is the minimal requirement for stimulating Pol\u03b8-pol MMEJ on relatively long ssDNA substrates, this indicates close cooperation between these two domains on long ssDNA. To gain insight into how these linked enzymes cooperate, we modeled the structure of the minimal end-joining Pol\u03b8\u0394cen protein on long ssDNA which combines the crystal structures of Pol\u03b8-pol and Pol\u03b8-hel attached by a flexible linker Fig.\u00a013,16. Ou13, we further investigated the footprint of Pol\u03b8\u0394cen on partial ssDNA (pssDNA) of similar size following incubation in MMEJ buffer conditions lacking dNTPs and BSA. Without ssDNA, 81% of Pol\u03b8 behaves as monomers and dimers, whereas 3% correspond to large (\u2009>\u20091000 \u00c5) multimers connected by a flexible linker Fig.\u00a0. The sphely Fig.\u00a0. Despiteely Fig.\u00a0\u20138.Fig. 731. The highly irregular shape of the assemblies suggests that, rather than droplets, the proteins form extended percolating networks; moreover, the fact that they are catalytically active indicates that the assemblies are permeable to the solvent and, possibly, large solutes. Based on the computational and EM evidence, we surmise that Pol\u03b8, and especially Pol\u03b8\u0394cen, form large (\u2009>\u2009100\u2009nm) gel-like complexes similar to others reported33, which accumulate DNA due to the high local concentration and accessibility of the polymerase and helicase domains.Overall, there is a close match between the morphology of the simulated complexes and the EM images Fig.\u00a0. A peculUsing a structure function approach in vitro, we investigated how recombinant human full-length Pol\u03b8 promotes MMEJ. On short ssDNA and short 3\u2032 overhangs, Pol\u03b8-pol efficiently performs MMEJ due to its interstrand pairing activity Fig.\u00a0. Thus, PPol\u03b8-hel stimulates Pol\u03b8-pol end-joining independently from its ATPase function. This supports the notion that Pol\u03b8-hel upregulates Pol\u03b8-pol MMEJ on long overhangs by simply binding ssDNA 5\u2032 proximal to the polymerase where it suppresses unproductive intrastrand pairing and snap-back replication by Pol\u03b8-pol Fig.\u00a0. Pol\u03b8-he34.Pol\u03b8 is unable to perform MMEJ or ssDNAx on short (\u2264\u200926\u2009nt) ssDNA Fig.\u00a0. HoweverPol\u03b8 forms multimeric complexes upon binding DNA. Replacement of Pol\u03b8-cen with a short linker, however, results in multimerization of the enzyme into an extended gel-like phase, even without DNA. This suggests that Pol\u03b8-cen suppresses Pol\u03b8 oligomerization by masking protein-protein interactions. Pol\u03b8 DNA binding may alter the conformation of Pol\u03b8-cen which in turn enables Pol\u03b8 oligomerization via protein-protein interactions. Our imaging data demonstrate that Pol\u03b8 multimers facilitate DNA accumulation and MMEJ. In summary, this report reveals the importance of the unique polymerase-helicase architecture of Pol\u03b8 which is essential for MMEJ.32P radiolabeled ssDNA in buffer ; the reaction was initiated by the addition of the indicated Pol\u03b8 enzyme and was incubated for 45\u2009min or as indicated at 37\u2009\u00b0C. For analysis in nondenaturing gels, reactions were terminated by the addition of non-denaturing stop buffer and incubated at 37\u2009\u00b0C for at least 15\u201320\u2009min. DNA was resolved in non-denaturing 11% or 12% polyacrylamide gels and analyzed by phosphorimager (Fujifilm FLA 7000). For time course experiments an aliquot of sample was removed from pooled reactions at the specified time point and transferred to tubes containing non-denaturing stop buffer. All quantified experiments were performed in triplicate and plotted with\u2009\u00b1\u2009s.d. Quantification was performed using ImageJ Gel Analysis. For XmaI digestion assays, after initial incubation with the indicated Pol\u03b8 enzyme the following buffer was added . 25 units of XmaI (New England Biolabs) was then added, as indicated, and incubated overnight at 37\u2009\u00b0C. The reaction was then stopped using non-denaturing stop buffer and resolved as above.10\u2009nM 5\u2032-32P radiolabeled pssDNA in buffer ; the reaction was initiated by the addition of the indicated Pol\u03b8 enzyme and incubated for 30\u2009min at 37\u2009\u00b0C. For analysis in denaturing gels, reactions were terminated by the addition of denaturing stop buffer (90% formamide and 50\u2009mM EDTA). DNA was resolved in denaturing 15% polyacrylamide gels and analyzed by phosphorimager (Fujifilm FLA 7000).10\u2009nM 5\u2032-32P) ATP and 100\u2009nM ssDNA (29 nt poly-dT) in buffer at room temp for the indicated times. The reaction mixture was then spotted onto a TLC plate on PEI cellulose, which was developed in a buffer containing 1\u2009M acetic acid and 0.25\u2009M LiCl2 for 1.5\u2009h. Plates were dried, then visualized by phosphorimager (Fujifilm FLA 7000).The indicated amounts of proteins were incubated with 10\u2009\u03bcM ATP, 2\u2009\u03bcCi of (\u03b3-: 5\u2009nM 5\u2032-32P radiolabeled pssDNA (RP538/RP539) in buffer was incubated with or without indicated Pol\u03b8 enzyme for 15\u2009min at 37\u2009\u00b0C. After initial incubation, 4 units of EcoRI were added as indicated and incubated for 10\u2009min at 37\u2009\u00b0C. Reactions were terminated by the addition of non-denaturing stop buffer and incubated at 37\u2009\u00b0C for at least 15\u201320\u2009min. DNA was resolved in non-denaturing 15% polyacrylamide gels and analyzed by phosphorimager (Fujifilm FLA 7000).T5 exonuclease assay: 8\u2009nM 3\u2032-Cy3 ssDNA (RP540Cy3) in buffer was mixed with 10\u2009nM Pol\u03b8\u0394cen or Pol\u03b8-pol and was preincubated for 10\u2009min at 37\u2009\u00b0C. After initial incubation, T5 Exonuclease (New England Biolabs) to a final concentration of 0.1\u2009U/ul or 0.5\u2009U/ul was added to the reactions as indicated and incubated for 15\u2009min at 37\u2009\u00b0C. Reactions were terminated by the addition of denaturing stop buffer (90% formamide and 50\u2009mM EDTA). DNA was resolved in denaturing 15% polyacrylamide gels and imaged at the Cy3 wavelength using FluorChem Q imager (Alpha Innotech). EcoRI endonuclease assay2, 2\u2009mM ATP, 30\u2009mM NaCl and 0.01% NP-40. Reactions comparing foci formation frequency were incubated for 10\u2009min at room temp before imaging, excluding time course reactions which were imaged as indicated. Cy3 foci were identified using a Leica DMi8 scanning confocal microscope with a 63x objective at the Cy3 emission wavelength. Random fields were collected for each condition and quantified using ImageJ particle analysis. Time course reactions were imaged at a single fixed field and quantified over time using ImageJ particle analysis of each individual frame. End-joining reactions performed with confocal imaging were performed as written with the addition of 20\u2009\u00b5M dNTPs. After confocal imaging, reactions were terminated by the addition of nondenaturing stop buffer and incubated at 37\u2009\u00b0C for 20\u2009min. Reactions were resolved in non-denaturing 12% polyacrylamide gels and imaged at the Cy3 wavelength using FluorChem Q imager (Alpha Innotech).Glass bottom wells were coated with 30\u2009mg/ml BSA for 30\u2009min at room temp. For foci formation frequency comparisons 40\u2009nM of Cy3 labeled DNA (RP334Cy3) was incubated with 3\u2009nM of the indicated Pol\u03b8 enzyme. For Rad52 confocal experiments 100\u2009nM Cy3 labeled DNA (RP334Cy3) was incubated with the indicated amounts of Rad52. For time course confocal assays and end-joining confocal assays 15\u2009nM of the indicated Pol\u03b8 enzyme was incubated with 40\u2009nM DNA (RP344Cy3). Reactions were performed with 25\u2009mM Tris-HCl, pH 8.8, 1\u2009mM DTT, 0.1\u2009mg/ml BSA, 10% glycerol, 10\u2009mM MgCl2, 10% glycerol, 0.1\u2009mg/ml BSA, 1\u2009mM ATP, 30\u2009mM NaCl for at least 30\u2009min at room temp. Reactions contained 10\u2009nM FAM-conjugated ssDNA (RP316FAM5), and the indicated amounts of the indicated Pol\u03b8 enzyme. A Clariostar (BMG Labtech) plate reader was used to measure fluorescence anisotropy. All experiments were performed in triplicate, normalized, and plotted with\u2009\u00b1\u2009s.d.Binding reactions were performed at room temp in 25\u2009mM Tris\u2013HCl, pH 8.8, 1\u2009mM DTT, 0.01% NP-40, 10\u2009mM MgCl2, 10% glycerol, 0.1\u2009mg/ml BSA, 1\u2009mM ATP, 30\u2009mM NaCl for at least 1\u2009hr at room temp. Reactions contained 10\u2009nM each of RP540Cy3 and RP541Cy5 and the indicated Pol\u03b8 enzyme. A Clariostar (BMG Labtech) plate reader was used to measure FRET . All experiments were performed in triplicate, normalized, and plotted with\u2009\u00b1\u2009s.d.Binding reactions were performed at room temp in 25\u2009mM Tris-HCl, pH 8.8, 1\u2009mM DTT, 0.01% NP-40, 10\u2009mM MgCl2, 2\u2009mM ATP, 0.01% NP-40 and 1\u2009mM DTT. Incubations were carried out at 37\u2009\u00b0C for 30\u2009min and deposited onto freshly cleaved mica. After 15\u2009s the mica surface was washed with milli Q water and dried with a stream of filtered air. Images were obtained on a NanoScope IV SFM operating in tapping mode in air with a type J scanner. Silicon Nanotips were from AppNano . Images were collected at 3\u2009\u03bcm\u2009\u00d7\u20093 \u03bcm and flattened to remove background slope using Nanoscope software. The size of proteins was measured from NanoScope images imported into IMAGE SXM 1.89 . Statistical analysis was done using QtiPlot (Version 0.9.8.9 svn 2288) and LibreOffice (Version: 5.1.6.2). The volume of proteins monomers, measured from SFM images of proteins deposited in 10\u2009mM Tris\u2013HCl, pH 8.8/100\u2009mM KCl, were 100\u2009nm3 for Pol\u03b8-hel and Pol\u03b8-pol, 210\u2009nm3 for Pol\u03b8\u0394cen and 320 nm3 for Pol\u03b8 and Pol\u03b8K121M. These values (\u2009\u00b1\u200950%) were used to normalize the volume of observed complexes. Quantification is presented as % of total proteins present in certain oligomeric state.Proteins (10\u2009nM) with or without ssDNA (20\u2009nM) were incubated in MMEJ buffer without dNTPs or BSA containing: 25\u2009mM Tris\u2013HCl, pH 8.8, 30\u2009mM NaCl or 100\u2009mM NaCl where indicated, 10\u2009mM MgCl2, 2\u2009mM ATP) at 37\u2009\u00b0C for 30\u2009min. Samples (4\u2009\u03bcl) were applied to glow discharged lacey carbon grids and incubated for 2\u2009min at room temp. The grids were then washed with water three times followed by staining with 2% uranyl acetate for 10\u2009s. Excess staining was removed by filter paper. The grids were air dried and imaged in a FEI Tecnai BioTwin Spirit transmission electron microscopy at Penn State University Cryo-Electron Microscopy facility.Pol\u03b8 or Pol\u03b8\u0394cen (200\u2009nM) with or without ssDNA (2\u2009\u03bcM) was incubated in 40\u2009\u03bcl of MMEJ reaction buffer without dNTPs, BSA or detergent or ATCC 208289/BJ5465 (a ura3-52 trp1 leu2-delta1 his3-delta200 pep4::HIS3 prb1-delta1.6\u2009R can1 GAL\u2009+\u2009), as previously described35. Colonies were picked and grown in SC-TRP with 2% raffinose at 30\u2009\u00b0C. Starter cultures were expanded and grown to an OD600 of 0.6\u20131.0. Expanded cultures were grown to an OD600 of 1.6\u20132, diluted with YPR to OD600 of 0.8\u20131.0 and induced as follows. Expression was induced by the addition of 2% galactose for 5\u2009hr at 30\u2009\u00b0C. Cells were harvested by centrifugation at 6,000\u2009rpm and washed with 50\u2009mM HEPES pH 8.0 and 1\u2009M Sorbitol then washed with lysis buffer . Cells were crushed in a freezer mill with liquid nitrogen. Frozen cell powder was stored at \u221280\u2009\u00b0C until purification. Frozen cell powder was thawed and resuspended in lysis buffer. The resuspended cell powder was centrifuged at 92,000\u2009g at 4\u2009\u00b0C for 30\u2009min. The clarified supernatant was re-centrifuged at 256,000\u2009\u00d7\u2009g at 4\u2009\u00b0C for 1\u2009h. The supernatant was collected. Anti-FLAG M2 resin was washed with 0.1\u2009M Glycine-HCl, pH 3.5; then with TBS buffer pH 7.4 then with lysis buffer by centrifugation at 1000\u2009rpm at 4\u2009\u00b0C for 5\u2009min. In total 5\u2009\u00b5g/ml 3xFLAG peptide (Sigma) and equilibrated anti-FLAG M2 resin (Sigma) were added to each tube and incubated at 4\u2009\u00b0C. The resin was settled by centrifugation at 1,000\u2009rpm at 4\u2009\u00b0C for 5\u2009min and flow-through was collected. The resin was washed with lysis buffer and then with wash buffer A . The resin was incubated in wash buffer A on ice for 15\u2009min, then the resin was settled by centrifugation at 1,000\u2009rpm at 4\u2009\u00b0C for 5\u2009min. The resin was then washed with wash buffer B . The resin was resuspended in elution buffer with 500\u2009\u00b5g/ml 3xFLAG peptide (Sigma) at 4\u2009\u00b0C with rotation. A disposable 10\u2009ml polypropylene column (Thermo Fisher) was washed with TBS buffer, pH 7.4 and then with elution buffer. The resin was loaded to the column and elution fractions were collected. The elution was dialyzed against dialysis buffer overnight at 4\u2009\u00b0C. Pol\u03b8, Pol\u03b8K121M and Pol\u03b8\u0394cen concentrations were determined by SDS gel analysis and by specific activity using Pol\u03b8-pol as a standard. Pol\u03b8, Pol\u03b8K121M and Pol\u03b8\u0394cen were stored in aliquots at \u221280\u2009\u00b0C.Pol\u03b8-pol, Pol\u03b8-hel and RPA were purified as describedTemplates are as follows: Figs.\u00a032P-5\u2032-radiolabeled with T4 polynucleotide kinase (New England Biolabs) and (\u03b3-32P) ATP (Perkin Elmer).\u00a0RP469D: CTGTCCTGCATGATG;RP486: CACTGTGAGCTTAGTCACATTTCATCATGCAGGACAG;RP344: CACTGTGAGCTTAGGGTTAGCCCGGG;RP348: CACTGTGAGCTTAGGGTTAGAGCCGG;SJB108: GTTCTTCGGTCTCGAGGTGACTACAAGGATGACGACGACAAGGGCACTGTGAGCTTAGGGTTAGCCCGGG;SJB116: CACTGTGAGCTTAGGGTTAGGCGGCTTGCAGAGCACAGAGGCCGCAGAATGTGCTCTAGATTCCGATGCTGACTTGCTGGGTATTATATGTGTGCCCGGG;RP514: GTTCTTCGGTCTCGAGGTGACTACAAGGATGACGACGACAAGGGCACTGTGAGCTTAGGGTTAGAAATTT;RP515: GTTCTTCGGTCTCGAGGTGACTACAAGGATGACGACGACAAGGGCACTGTGAGCTTAGGGTTAGCCGG;RP132: GACGTTGACTTAAAGTCTAACCTATAGGATACTTACAGCCATCGAGAGGGACACGGCGCATTCTCGAGCGTAC;RP132C: GCTCGAGAATGCGCCGTGTCCCTCTCGATGGCTGTAAGTATCCTATAGGTTAGACTTTAAGTCAACGTCGTAC;RP370: CACTGTGAGCTTAGGGTTAGGAATTC;RP316FAM5: /56-FAM/TTTTTTTTTTTTTTTTTTTTTTTTTTTTT;SJB123: GGTTAGCCCGGG;RP344Cy3: /5Cy3/TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT;RP343: CTAAGCTCACAGTG;SJB153: TATAATACCCAGCAAGTCAGCATC; SJB154: GGAATCTAGAGCACATTCTGCGGCC;SJB155: TCTGTGCTCTGCAAGCCGCCTAACC;SJB158: TATAATACCCAGCAAGTCAGCAT/3ddC/;SJB159: GGAATCTAGAGCACATTCTGCGGC/3ddC/;RP540Cy3: GCATATTCACTGTGAGCTTAGTGTTAGGACTCGG/3Cy3Sp/;RP541Cy5: GCATATTCACTGTGAGCTTAGTGTTACCGAGTCC/3Cy5Sp/;RP538: CGACAAGAGTCATGAATTCTTAGGGTTAGCCCGGG;RP539: CTAAGAATTCATGACTCTTGTCG;SJB089: CAATTCAGCAACTAATGTCATACCAGCTGAAGTTGGTGCAGAG;SJB090: CTCTGCACCAACTTCAGCTGGTATGACATTAGTTGCTGAATTG;SJB100: TTCTGCTAGCGGTGGTGGAGGAAGTGGAGGAGGCGGATCTGGTGGTGGCGGTAGCGGTTTTAAAGATAACTCTCCAATCTCAGATACTTC;SJB101: TCTTCTGCTAGCCATTTCAACCAAATCTTGTTGCAAG.Primer templates were 5\u2032- phosphorylated on the shorter strand with T4 polynucleotide kinase (New England Biolabs) and ATP. Primer templates were annealed by mixture of a ratio of 1:2 of short to long strands then boiling and slow cooling to room temp. DNA was 3xFLAG-Pol\u03b8 plasmid was derived from pRS424 (ATCC). Human Pol\u03b8 nucleotide sequence was optimized for yeast and synthesized as 2 gene fragments by GenScript. Yeast optimized gene fragments were cloned into pRS424 sequentially. 3xFLAG-Pol\u03b8K121M plasmid was derived from 3xFLAG-Pol\u03b8 plasmid by site-directed mutagenesis using primers (SJB089 and SJB090). 3xFLAG-Pol\u03b8\u0394cen was derived from 3xFLAG-Pol\u03b8 plasmid by molecular cloning with PCR primers (SJB100 and SJB101).Pol\u03b8, Pol\u03b8-pol, and Pol\u03b8\u0394cen were resolved by SDS/PAGE and transferred to a nitrocellulose membrane. The membrane was blocked with 5% milk in TBS with 0.1% Tween-20 and incubated with primary antibody against C-terminal portion of Pol\u03b8 in TBS/5% milk/0.1% Tween-20 for 1.5\u2009hr at room temp, followed by incubation with HRP-conjugated secondary antibody for 1\u2009h at room temp. Bands were detected via chemiluminescence.13 and Pol\u03b8-pol 16. The Pol\u03b8-hel:ssDNA model was constructed by superposing Pol\u03b8-hel structure with the crystal structure of archaeal DNA helicase (Hel308):DNA complex28, and the strand in which Hel308 translocates on was included in the model. The template DNA in the Pol\u03b8-pol:DNA complex was removed and the 5\u2032 end of the primer DNA was connected to the 3\u2032-end of DNA in the Pol\u03b8-hel model. Pol\u03b8-hel was modeled facing upstream due to its 3\u2032\u22125\u2032 translocation activity.The Pol\u03b8\u0394cen:ssDNA model was constructed by using the previously determined crystal structures of human Pol\u03b8-hel 3 linker (L), see Supplementary Fig.\u00a0Pol\u03b8\u0394cen is modelled by two large spheres, representing the polymerase (P) and helicase (H) domains, connected by a short flexible polymer chain made by 6 beads, representing the (GGGGS)T\u2009=\u2009300\u2009K and with a potential energy V, composed by the following interaction terms.Each bead is governed by a Langevin equation of motion at temp VP-P(r)\u2009=\u2009VH-H(r)\u2009=\u2009\u03b5((\u03c3/r)12-(\u03c3/r)6), where \u03c3\u2009=\u20095.6\u2009nm is about twice the gyration radius of both P and H and \u03b5\u2009=\u200910KBT is the interaction strength. The potentials VP-P and VH-H are truncated at r\u2009=\u20098\u2009nm, which correspond to a cutoff distance of about 1.5\u2009nm from their potential minimum, situated at 21/6\u03c3. The distance 1.5\u2009nm corresponds roughly to the Debye screening length for this system.The interactions between polymerase-polymerase and helicase-helicase are attractive, and are modelled with a Lennard-Jones potential r\u2009=\u200921/6\u03c3.All other bead-bead interactions are instead only repulsive and modelled with a Lennard\u2013Jones potential truncated at the potential minimum VP-H(r)), \u03c3\u2009=\u20095.6\u2009nm and \u03b5\u2009=\u200910KBT. For linker-linker interactions (VL-L(r)), \u03c3\u2009=\u20091\u2009nm, corresponding to about 2.5 amino acids, and \u03b5\u2009=\u20091KBT. For polymerase-linker and helicase-linker (VP-L(r)\u2009=\u2009VH-L(r)) \u03c3\u2009=\u20093.3\u2009nm and \u03b5\u2009=\u200910KBT.\u03c3 and \u03b5 vary depending on the particle types. For polymerase-helicase interactions (VH\u2009=\u2009K(r-r0), where K\u2009=\u2009750 KBT/nm2 and r0\u2009=\u20091\u2009nm for linker-linker bonds and r0\u2009=\u20093.3\u2009nm for linker-helicase and linker-polymerase bonds.The connectivity between the beads of each Pol\u03b8\u2206cen model , and the size range of the simulation boxes allow to reproduce the local concentration, size and shape of multimers observed in the EM data.To generate the assemblies of varying size, we initialized the system with proteins randomly distributed at densities of about 1\u20133\u2009\u00d7\u200910BT and m\u2009=\u2009900\u2009Da. In simulation units, the integration timestep is dt\u2009=\u20090.01 and the friction coefficient is \u03b3\u2009=\u20090.5. The simulation box is bounded by repulsive walls with a repulsive truncated Lennard\u2013Jones potential, where the truncation is set at r\u2009=\u200921/6\u03c3. A typical simulation is long 5\u20136\u2009\u00d7\u2009106 time units.The simulations are carried out with the software LAMMPS with reduced units \u03c3\u2009=\u20091\u2009nm, \u03b5\u2009=\u20091KThe clusters generated by the self-assembly process have a peculiar elongated shape, which is also characteristic of the EM images. This structural feature is absent if the helicase-helicase or polymerase-polymerase attractive interactions are substituted with a truncated repulsive Lennard-Jones potential, see Supplementary Fig.\u00a0Further information on research design is available in the\u00a0Supplementary InformationPeer Review FileDescription of Additional Supplementary FilesSupplementary Movie 1Supplementary Movie 2Supplementary Movie 3Supplementary Movie 4Supplementary Movie 5Supplementary Movie 6Supplementary Movie 7Supplementary Movie 8Reporting SummarySource Data"} +{"text": "Nature Communications 10.1038/s41467-018-03728-5; published online: 10 April 2018Correction to: The originally published version of this article contained an error in the name of the author Fl\u00f3ra G\u00f6l\u00f6ncs\u00e9r, which was incorrectly given as Fl\u00f3ra G\u00f6r\u00f6ncs\u00e9r. This has now been corrected in both the PDF and HTML versions of the article."} +{"text": "In the title com\u00adpound, inter\u00admolecular C\u2014H\u22efO, O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular network. 13H14N2O3, the dihydropyridazine ring (r.m.s. deviation\u00a0= 0.166\u2005\u00c5) has a screw-boat conformation. The dihedral angle between its mean plane and the benzene ring is 0.77\u2005(12)\u00b0. In the crystal, inter\u00admolecular O\u2014H\u22efO hydrogen bonds generate C(5) chains and N\u2014H\u22efO hydrogen bonds produce R22(8) motifs. These types of inter\u00adactions lead to the formation of layers parallel to (12R24(8) motifs. Inter\u00admolecular inter\u00adactions were additionally investigated using Hirshfeld surface analysis and two-dimensional fingerprint plots. The most significant contributions to the crystal packing are by H\u22efH (43.3%), H\u22efC/C\u22efH (19.3%), H\u22efO/H\u22efO (22.6%), C\u22efN/N\u22efC (3.0%) and H\u22efN/N\u22efH (5.8%) contacts. C\u2014H\u22ef\u03c0 inter\u00adactions and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions are not observed.In the title com\u00adpound, C This nitro\u00adgen heterocycle became a scaffold of choice for the development of potential drug candidates -6-(4-hy\u00addroxy-3-meth\u00adoxy\u00adstyr\u00adyl)-4,5-di\u00adhydro\u00adpyridazin-3(2H)-one, as well as an analysis of the Hirshfeld surfaces.In continuation of our studies related to mol\u00adecular structures and Hirshfeld surface analysis of new heterocyclic derivatives \u2005\u00c5 for the C3 atom; the C2 atom lies \u22120.177\u2005(3)\u2005\u00c5 out of the plane in the opposite direction relative to the C3 atom. The dihedral angle between the dihydropyridazine ring mean plane and the benzene ring (C7\u2013C12) is 0.77\u2005(12)\u00b0, indicating an almost planar conformation of the molecule favouring delocalization over the C4\u2014C5=C6\u2014C7 bridge.In the title mol\u00adecule Fig.\u00a01, the coni hydrogen bonds between the phenolic OH group and the carbonyl O atom of a neighbouring mol\u00adecule generate C(5) chains extending parallel to [101]. Likewise, N1\u2014H1\u22efO1ii hydrogen bonds between the N\u2014H function of the di\u00adhydro\u00adpyridazine ring and the carbonyl O atom generate centrosymmetric dimers with an A\u22efO2iii and C13\u2014H13C\u22efO2iv hydrogen bonds with In the crystal, mol\u00adecules are stacked in rows parallel to [100]. Notably, no significant C\u2014H\u22ef\u03c0 or \u03c0\u2013\u03c0 inter\u00adactions are observed. O2\u2014H2\u22efO1fs Fig.\u00a02.et al., 2016viz. 6-phenyl-4,5-di\u00adhydro\u00adpyridazin-3(2H)-one -(\u2212)-6-(4-amino\u00adphen\u00adyl)-5-methyl-4,5-di\u00adhydro\u00adpyridazin-3(2H)-one to 2.2382\u2005a.u. (blue). The surfaces mapped over relevant inter\u00admolecular contacts are illustrated in Fig.\u00a03dnorm plotted onto the surface are shown for the H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH, C\u22efN/N\u22efC and H\u22efN/N\u22efH inter\u00adactions in Figs.\u00a04a)\u2013(e), respectively. The overall two-dimensional fingerprint plot and those delineated into H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH, C\u22efN/N\u22efC and H\u22efN/N\u22efH contacts are illustrated in Figs.\u00a05a)\u2013(f), respectively. The largest inter\u00adaction is that of H\u22efH, contributing 43.3% to the overall crystal packing. H\u22efC/C\u22efH contacts add a 19.3% contribution to the Hirshfeld surface, with the tips at de + di \u223c 2.72\u2005\u00c5. H\u22efO/O\u22efH contacts make a 22.6% contribution to the Hirshfeld surface and are represented by a pair of sharp spikes in the region de + di\u00a0\u223c 2.70\u2005\u00c5 in the fingerprint plot. H\u22efO/O\u22efH inter\u00adactions arise from inter\u00admolecular O\u2014H\u22efO hydrogen bonding and C\u2014H\u22efO contacts. The contributions of the other contacts to the Hirshfeld surface are negligible, i.e. C\u22efN/N\u22efC of 3.0% and H\u22efN/N\u22efH of 5.8%.Hirshfeld surface analysis was used to qu\u00adantify the inter\u00admolecular inter\u00adactions of the title com\u00adpound, using To a solution of 6-(4-hy\u00addroxy-3-meth\u00adoxy\u00adphen\u00adyl)-4-oxohex-5-enoic acid in 20\u2005ml of ethanol, an equimolar amount of hydrazine hydrate was added. The mixture was maintained under reflux until thin-layer chromatography (TLC) indicated the end of the reaction. After cooling, the precipitate which formed was filtered off, washed with ethanol and recrystallized from ethanol. Slow evaporation at room temperature led to the formation of single crystals of the title com\u00adpound.Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) otherwise. The NH and OH hydrogens were located in a difference Fourier map and were constrained with N\u2014H = 0.86\u2005\u00c5 and Uiso(H) = 1.2Ueq(N), and O\u2014H = 0.86\u2005\u00c5 and Uiso(H) = 1.5Ueq(O), using a riding model.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019014130/wm5521sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019014130/wm5521Isup3.hklStructure factors: contains datablock(s) I. DOI: 1959568, 1959568CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-thio\u00adchromene (A) and 2-(2-bromo-5-fluoro\u00adphen\u00adyl)-7-meth\u00adoxy-3-nitro-2H-thio\u00adchromene (B) are described. In each crystal, the mol\u00adecules are linked by hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions.The synthesis and structure of 2-(2-bromo-5-fluoro\u00adphen\u00adyl)-8-eth\u00adoxy-3-nitro-2 H-thio\u00adchromene and 2-(2-bromo-5-fluoro\u00adphen\u00adyl)-7-meth\u00adoxy-3-nitro-2H-thio\u00adchromene , were prepared via the condensation reaction between 2-mer\u00adcapto\u00adbenzaldehyde and nitro\u00adstyrene derivatives. In both com\u00adpounds, the thio\u00adchromene plane is almost perpendicular to the phenyl ring. In the structure of A, mol\u00adecules are assembled via \u03c0\u2013\u03c0 stacking and C\u2014H\u22efO and C\u2014F\u22ef\u03c0 inter\u00adactions. In the crystal packing of B, mol\u00adecules are linked by C\u2014H\u22efF, C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions.Two thio\u00adchromene com\u00adpounds containing Br and F atoms, namely 2-(2-bromo-5-fluoro\u00adphen\u00adyl)-8-eth\u00adoxy-3-nitro-2 H-Chromenes (or 2H-benzo\u00adpyrans) are heterocyclic com\u00adpounds found in many natural plants. This class of mol\u00adecules shows a wide variety of biological activities, such as anti\u00adcancer, anti-inflammation and anti-HIV and 2-(2-bromo-5-fluoro\u00adphen\u00adyl)-7-meth\u00adoxy-3-nitro-2H-thio\u00adchromene (B). Crystal structure determination can help to understand the role of halogenated substituents in the biological activity of these com\u00adpounds.2A) crystallizes in the triclinic space group PB) crystallizes in the space group P21/c, both with one mol\u00adecule in the asymmetric unit \u00b0 with phenyl ring C1\u2013C6, while in B, this angle is 86.94\u2005(8)\u00b0, which indicates that the 2-bromo-5-fluoro\u00adphenyl ring is roughly perpendicular to the thio\u00adchromene plane. Both 2H-thio\u00adpyran rings have a screw-boat conformation, with atom C7 having the largest deviation from the best plane through atoms S1/C7\u2013C11 . The C\u2014S bond lengths are almost equal . The C11\u2014S1\u2014C7 bond angle is 102.5\u2005(2)\u00b0 in A and 100.47\u2005(9)\u00b0 in B. The N\u2014O bond lengths in com\u00adpound B [1.232\u2005(2) and 1.221\u2005(2)\u2005\u00c5] are slightly longer than those in com\u00adpound A [both 1.219\u2005(5)\u2005\u00c5]. The nitro group is situated in the thio\u00adchromene plane, as illustrated by the torsion angle O2\u2014N8\u2014C8\u2014C9 of 1.3\u2005(7)\u00b0 in A and 9.5\u2005(3)\u00b0 in B.Compound \u2005\u00c5; symmetry code: (i) \u2212x, \u2212y\u00a0+\u00a02, \u2212z; Cg3 is the centroid of the C10\u2013C15 ring]. Neighbouring dimers inter\u00adact through C\u2014F\u22ef\u03c0 and short Br1\u22efH5ii inter\u00adactions .In the crystal of s Table\u00a01 and \u03c0\u2013\u03c0 B, two mol\u00adecules form dimers through C\u2014H\u22efF hydrogen bonds (Table\u00a02c direction through \u03c0\u2013\u03c0 inter\u00adactions . Parallel chains inter\u00adact via C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions.In the crystal of com\u00adpound s Table\u00a02. These det al., 2016H-thio\u00adchromene derivatives, of which three contain halogen atoms [CSD refcodes IFOZIO in toluene and the reaction mixture was stirred at room temperature for 2\u2005h. After com\u00adpletion of the reaction, the solvent was evaporated under reduced pressure and the crude product was purified by flash chromatography on silica gel (yield 90%). Crystals suitable for single-crystal X-ray diffraction data collection were obtained by slow evaporation from an ethanol solution.To a round-bottomed flask was added 2-mercaptobenzaldehyde (1 equiv.), nitro\u00adstyrene (1 equiv.) and KUiso(H) = 1.2Ueq(C) for Csp2\u2014H and Uiso(H) = 1.5Ueq(C) for Csp3\u2014H. A rotating-group model was applied for methyl-group C17 in A and C16 in B.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019014178/vm2223sup1.cifCrystal structure: contains datablock(s) A, B, global. DOI: 10.1107/S2056989019014178/vm2223Asup8.hklStructure factors: contains datablock(s) A. DOI: 10.1107/S2056989019014178/vm2223Bsup9.hklStructure factors: contains datablock(s) B. DOI: Click here for additional data file.10.1107/S2056989019014178/vm2223Asup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019014178/vm2223Bsup5.cmlSupporting information file. DOI: 1950406, 1953121, 1950406, 1953121CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound has a nonplanar conformation. In the crystal, the anions are linked to the cations and the water mol\u00adecules by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, forming a three-dimensional network. Face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions are also observed. 2H10N22+\u00b7C21H13N3O8S2\u2212\u00b72H2O, the planes of the phenyl rings and the benzene ring of the 5-nitro-2-oxido\u00adbenzene\u00adsulfonate group are inclined to one another by 44.42\u2005(11), 56.87\u2005(11) and 77.70\u2005(12)\u00b0. In the crystal, the anions are linked to the cations and the water mol\u00adecules by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, forming a three-dimensional network. Furthermore, there are face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions between the centroids of one phenyl ring and the benzene ring of the 5-nitro-2-oxido\u00adbenzene\u00adsulfonate group [centroid\u2013centroid distance = 3.8382\u2005(13)\u2005\u00c5 and slippage = 1.841\u2005\u00c5]. A Hirshfeld surface analysis was conducted to verify the contributions of the different inter\u00admolecular inter\u00adactions.In the anion of the title hydrated salt, C DOI: 10.1107/S2056989018009118/qm2125Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018009118/qm2125Isup3.cmlSupporting information file. DOI: 1851087CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, inter\u00admolecular N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional supra\u00admolecular network. H-pyridazin-3-one derivative. The mol\u00adecule, C18H16N2O, is not planar; the benzene and pyridazine rings are twisted with respect to each other, making a dihedral angle of 11.47\u2005(2)\u00b0, and the toluene ring is nearly perpendicular to the pyridazine ring, with a dihedral angle of 89.624\u2005(1)\u00b0. The mol\u00adecular conformation is stabilized by weak intra\u00admolecular C\u2014H\u22efN contacts. In the crystal, pairs of N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into inversion dimers with an R22(8) ring motif. The inter\u00admolecular inter\u00adactions were investigated using Hirshfeld surface analysis and two-dimensional (2D) fingerprint plots, revealing that the most important contributions for the crystal packing are from H\u22efH (56.6%), H\u22efC/C\u22efH (22.6%), O\u22efH/H\u22efO (10.0%) and N\u22efC/C\u22efN (3.5%) inter\u00adactions.In this paper, we describe the synthesis of a new di\u00adhydro-2 Pyridazinone is an important pharmacophore possessing a wide range of biological applications . Weak C\u2014H\u22efO hydrogen bonds and weak off-set \u03c0-stacking stabilize the packing. In the crystal, hydrogen bonds link the chains into a two-dimensional (2D) network parallel to (011) ], with a centroid-to-centroid distance of 3.8333\u2005(18)\u2005\u00c5 and a slippage of 1.460\u2005\u00c5 (Cg1 is the centroid of the C9\u2013C11/N1/N2 ring and Cg3 is the centroid of the C13\u2013C18 ring) .The mol\u00adecules are connected two-by-two through N1\u2014H1\u22efO1 hydrogen bonds Table\u00a02, with a g) Fig.\u00a02a.et al., 2016H)-one (see A in Schemep-tolyl\u00adpyridazin-3(2H)-one to 1.4188\u2005a.u. (blue). The 3D dnorm surface of the title complex is illustrated in Figs.\u00a03a) and 4dnorm values on the surface correspond to the N\u2014H\u22efO inter\u00adactions is due to short inter\u00adatomic H\u22efH contacts. In the absence of C\u2014H\u22ef\u03c0 inter\u00adactions in the crystal, the pair of characteristic wings in the fingerprint plot representing H\u22efC/C\u22efH contacts (22.6% contribution to the HS) have a symmetrical distribution of points , with the tips at de\u00a0+ di = 2.797\u2005\u00c5. The O\u22efH contacts contribute 10% to the HS and have a symmetrical distribution of points, with the tips at de + di = 1.853\u2005\u00c5. The contribution of the other contact to the Hirshfeld surface is N\u22efC/C\u22efN (3.5%). The Hirshfeld surface representations with the function dnorm plotted on the surface are shown for the H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH, C\u22efC and H\u22efN/N\u22efH inter\u00adactions in Figs.\u00a06et al., 2015The Hirshfeld surface analysis . The convex blue regions on the shape-index symbolize hydrogen-donor groups and the concave red regions symbolize hydrogen-acceptor groups. The \u03c0\u2013\u03c0 inter\u00adactions on the shape-index map of the Hirshfeld surface are generally indicated by adjacent red and blue triangles.A shape-index map of the title compound was generated in the range \u22121 to 1\u2005\u00c5 Fig.\u00a03b. The cc). This shows large regions of green indicating a relatively flat surface area (planar), while the blue regions indicate areas of curvature. The presence of \u03c0\u2013\u03c0 stacking inter\u00adactions is also evident in the flat regions around the rings on the Hirshfeld surface plotted over curvedness (see the Supra\u00admolecular features section above).A curvedness map of the title compound was generated in the range \u22124 to 0.4\u2005\u00c5 Fig.\u00a03c. This H)-one and of 4-methyl\u00adbenz\u00adaldehyde in ethanol (30\u2005ml), sodium hydroxide was added. The solvent was evaporated under vacuum and the residue was purified through silica-gel column chromatography using hexa\u00adne/ethyl acetate (7:3 v/v). Slow evaporation at room temperature leads to single crystals.To a solution of 6-phenyl-4,5-di\u00adhydro\u00adpyridazin-3(2Uiso(H) = 1.5Ueq(C) for methyl, C\u2014H = 0.96\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for methyl\u00adene, C\u2014H = 0.93\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for aromatic and C\u2014H = 0.98\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for methine H atoms. Crystal data, data collection and structure refinement details are summarized in Table\u00a03H atoms were fixed geometrically and treated as riding, with C\u2014H = 0.97\u2005\u00c5 and 10.1107/S2056989019011551/mw2146sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019011551/mw2146Isup3.hklStructure factors: contains datablock(s) I. DOI: 1947718CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-indene-1,3(2H)-dione (ID[1]) and (E)-2-{3-[4-(di\u00admethyl\u00adamino)\u00adphen\u00adyl]allyl\u00adidene}-1H-indene-1,3(2H)-dione (ID[2]), with donor\u2013\u03c0-bridge\u2013acceptor structures, are almost planar for the mol\u00adecule with a short \u03c0-bridge (ID[1]) but less planar for the mol\u00adecule with a longer bridge (ID[2]).The title pull\u2013push chromophores, 2-[4-(di\u00admethyl\u00adamino)\u00adbenzyl\u00adidene]-1 H-indene-1,3(2H)-dione, C18H15NO2 (ID[1]) and (E)-2-{3-[4-(di\u00admethyl\u00adamino)\u00adphen\u00adyl]allyl\u00adidene}-1H-indene-1,3(2H)-dione, C20H17NO2 (ID[2]), have donor\u2013\u03c0-bridge\u2013acceptor structures. The mol\u00adecule with the short \u03c0-bridge, ID[1], is almost planar while for the mol\u00adecule with a longer bridge, ID[2], is less planar. The benzene ring is inclined to the mean plane of the 2,3-di\u00adhydro-1H-indene unit by 3.19\u2005(4)\u00b0 in ID[1] and 13.06\u2005(8)\u00b0 in ID[2]. The structures of three polymorphs of compound ID[1] have been reported: the \u03b1-polymorph is non-centrosymmetric (space group P21), which suggests potential NLO properties for this crystalline material. In both compounds, there is short intra\u00admolecular C\u2014H\u22efO contact present, enclosing an S(7) ring motif. In the crystal of ID[1], mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming layers parallel to the bc plane. In the crystal of ID[2], mol\u00adecules are liked by C\u2014H\u22efO hydrogen bonds to form 21 helices propagating along the b-axis direction. The mol\u00adecules in the helix are linked by offset \u03c0\u2013\u03c0 inter\u00adactions with, for example, a centroid\u2013centroid distance of 3.9664\u2005(13)\u2005\u00c5 (= b axis) separating the indene rings, and an offset of 1.869\u2005\u00c5. Spectroscopic and electrochemical measurements show the ability of these compounds to easily transfer electrons through the \u03c0-conjugated chain.The title pull\u2013push chromophores, 2-[4-(di\u00admethyl\u00adamino)\u00adbenzyl\u00adidene]-1kova 1978. Kristalkova 1980. Kristalkaya 1980. Kristal Applications of pull\u2013push chromophores include non-linear optics are important in many areas of materials chemistry, especially organic electronics and optoelectronics. Applic\u00adations of pull\u2013push mol\u00adecules can be related to their properties such as intra\u00admolecular charge transfer and specific mol\u00adecular arrangements in the solid state. Intra\u00admolecular charge transfer from donor to acceptor The mol\u00adecular structures of ID[1] and ID[2] are illustrated in Fig.\u00a01et al., 2019Both mol\u00adecules have acceptor\u2013\u03c0-bridge\u2013donor structures. It was found, as in our previous studies being inclined to the mean plane of the indene ring system (C1\u2013C9) by 3.19\u2005(4)\u00b0. In ID[2] the deviation from planarity is somewhat larger with the benzene ring (C10\u2013C15) being inclined to the mean plane of the indene ring system (C1\u2013C9) by 13.06\u2005(8)\u00b0; see further details in Table\u00a01Mol\u00adecules of ID[1] and ID[2] have significant dipole moments, which is very common for NLO chromophores. Because of this, mol\u00adecules have a trend to anti\u00adparallel packing, which is observed in the crystal structures of both ID[1] and ID[2].ca 84.47\u00b0 between them. The mol\u00adecules, which stack in an anti\u00adparallel or head-to-tail fashion, are linked by C\u2014H\u22efO hydrogen bonds \u2005\u00c5 separating the indene ring systems (C1\u2013C9), with an offset of 1.869\u2005\u00c5.In the crystal of ID[2], mol\u00adecules form stacks with parallel mol\u00adecular positions, and shifted positions of stacks extended along the on Fig.\u00a03. Here, ts Table\u00a03, forminget al., 2019Absorbance spectra were obtained for both ID[1] and ID[2] in chloro\u00adform and aceto\u00adnitrile. For donor\u2013acceptor polyenes, the dominating feature of the absorbance spectrum is the \u03c0\u2013\u03c0* transition that results from charge transfer from donor to acceptor. According to recent studies (Bogdanov a), however, when swept to \u22121.9\u2005V the reduction is only partly reversible . This represents the ability of the compound to \u2018easily\u2019 transfer electrons through the chain from donor towards acceptor.Donor\u2013acceptor polyenes can be characterized by electrochemical measurements to show their ability to transfer electrons. The voltammagrams Fig.\u00a05 demonstrle Fig.\u00a05b. This 2+/0 in di\u00adchloro\u00admethane with 0.1 nM Bu4NPF6). Measurements were recorded at 50\u2005mV\u2005s\u22121 using a BAS Potentiostat using a glassy carbon working electrode, Pt wire auxilliary electrode and a Ag/AgCl reference electrode.Note: cyclic voltammagrams of ID[1] were made against FeCpet al., 2016p-di\u00adethyl\u00adamino\u00adbenzyl\u00adidene)-1,3-indandione -dione -di\u00adone , 1.66\u2005(6)/1.49\u2005(9) and 5.71\u2005(9)\u00b0, for TELWEN, QENYEQ/QENYEQ01 and BIQYUY, respectively, compared to 3.19\u2005(4)\u00b0 in ID[1].A search of the Cambridge Structural Database , while in the \u03b3 polymorph it is acentric (space group Pna21). The crystal structure of ID[1] we obtained corresponds to the \u03b2 polymorph, i.e. the centrosymmetric modification MBYINO02. The dihedral angles between the benzene and indene rings for two independent mol\u00adecules in MBYINO are ca 4.35 and 7.79\u00b0, compared to ca 7.36\u00b0 in MBYINO01, and 3.54\u00b0 in MBYINO02 (cf. 3.19\u2005(4)\u00b0 in the present crystal structure analysis of ID[1]).Also, out of all 27 hits there are three hits, -dione \u00b0 in ID[2]. In ZIGPIR this dihedral angle is smaller at 8.6\u2005(3)\u00b0.A search of the CSD for the substructure of ID[2] yielded nine hits. Only two structures are similar to that of ID[2]. The first, 2-{3-[4-(di\u00admethyl\u00adamino)\u00adphen\u00adyl]prop-2-yn-1-yl\u00adidene}-1et al., 2019For the synthesis of the title compounds, two aldehydes were used: 4-(di\u00admethyl\u00adamino)\u00adbenzaldehyde and 4-(di\u00admethyl\u00adamino)\u00adcinnamaldehyde (A2), which was synthesized as described previously \u00adbenzyl\u00adidene]indane-1,3-dione (ID[1]): Aldehyde A1 and 1,3-indanedione were suspended in 100\u2005ml of absolute ethanol. The mixture was gently heated until the solids had dissolved. After about 10\u2005min of stirring the dissolution was complete, and a red crystalline precipitate began forming on the walls of the flask. The reaction mixture was stirred vigorously overnight, and the resulting product was collected by filtration then washed with cold ethanol and hexa\u00adnes to give shiny dark-red crystals . ID[1] can be purified by recrystallization using numerous solvent systems , many of which afforded single crystals. 1H NMR \u03b4 8.52 , 7.91\u20139.73 , 7.71 , 6.77 , 3.14 ppm. 13C NMR \u03b4 191.6, 190.1, 154.5, 147.3, 142.7, 140.3, 138.2, 134.8, 134.5, 123.3, 122.7, 122.6, 122.2, 111.7, 40.3 ppm.Synthesis of (E)-2-{3-[4-(di\u00admethyl\u00adamino)\u00adphen\u00adyl]allyl\u00adidene}indane-1,3-dione (ID[2]): Aldehyde A2 , 1,3-indanedione and piperidine in ethanol (50\u2005ml) were mixed and treated as for the synthesis of ID[1]. The crude product obtained was collected by filtration and washed with cold ethanol before being recrystallized from ethanol to give incredibly shiny and thin purple actinic crystals . They were washed with hexane and dried under vacuum. 1H NMR \u03b4 8.25 , 7.90\u20137.86 , 7.76\u20137.73 , 7.60 , 7.60 , 7.34 , 6.72 , 3.08 ppm.Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Uiso(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S205698901901329X/su5515sup1.cifCrystal structure: contains datablock(s) ID1, ID2, Gobal. DOI: 10.1107/S205698901901329X/su5515ID1sup2.hklStructure factors: contains datablock(s) ID1. DOI: 10.1107/S205698901901329X/su5515ID2sup3.hklStructure factors: contains datablock(s) ID2. DOI: Click here for additional data file.10.1107/S205698901901329X/su5515ID1sup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901901329X/su5515ID2sup5.cmlSupporting information file. DOI: 1956419, 1956418, 1956418, 1956419CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N\u2010Methylation of lysyl residues is widely observed on histone proteins. Using isolated enzymes, we report mechanistic and structural studies on histone lysine demethylase (KDM)\u2010catalysed demethylation of N\u03b5\u2010methylated lysine 26 on histone 1 isotype 4 (H1.4). The results reveal that methylated H1.4K26 is a substrate for all members of the KDM4 subfamily and that KDM4A\u2010catalysed demethylation of H1.4K26me3 peptide is similarly efficient to that of H3K9me3. Crystallographic studies of an H1.4K26me3:KDM4A complex reveal a conserved binding geometry to that of H3K9me3. In the light of the high activity of the KDM4s on this mark, our results suggest JmjC KDM\u2010catalysed demethylation of H1.4K26 may be as prevalent as demethylation on the H3 tail and warrants further investigation in cells. Our findings reveal that demethylation of methylated lysyl residues in H1.4K26 peptide fragments is catalysed by all members of the human KDM4 subfamily of JmjC KDMs (KDM4A\u2010E), supporting previously reported cell\u2010based studies (predominantly with mouse enzymes) Escherichia coli . Sequences are given in Table\u00a0Peptides were produced as C\u2010terminal amides using a Liberty Blue automated microwave peptide synthesiser , using standard fluorenylmethyloxycarbonyl\u2010mediated solid\u2010phase chemistry and NovaPEG rink amide resin . Following synthesis, peptides were cleaved from the resin by incubation with trifluoroacetic acid/water/triisopropylsilane/dimethoxybenzene (92.5 : 2.5\u00a0:\u00a02.5\u00a0: 2.5) for 3\u00a0h followed by precipitation with ice\u2010cold diethyl ether. For kinetic experiments, lyophilised peptides were purified by reverse\u2010phase high\u2010performance liquid chromatography using a Vydac C18 column (Solvent A: 0.1% trifluoroacetic acid in Hm) were incubated with peptides ammonium sulfate (10\u00a0\u03bcm), sodium ascorbate (100\u00a0\u03bcm) and 2\u2010oxoglutaric acid (200\u00a0\u03bcm). Reactions with KDM3A and KDM7A also contained 1\u00a0mm tris(2\u2010carboxyethyl)phosphine hydrochloride. No enzyme reactions were included as negative controls. Reactions were incubated for 1\u00a0h at 37\u00a0\u00b0C before being quenched with 1\u00a0:\u00a01 (v/v) methanol. Analysis was carried out by MALDI\u2010TOF MS , with demethylation observed as mass shifts of 14\u00a0Da.Recombinant proteins (1\u00a0\u03bc1H observation and were carried out as previously described m), 2OG (400\u00a0\u03bcm), H1.4(18\u201332)K26me3 peptide (400\u00a0\u03bcm), Fe(II) ammonium sulfate (100\u00a0\u03bcm) and ascorbate (1\u00a0mm) in 50\u00a0mm ammonium formate, pH 7.5, 500\u00a0mm NaCl in 10% v/v D2O) were prepared in a microcentrifuge tube , before being transferred to an NMR tube. The samples were then analysed over time using automated routines; each time\u2010point corresponded to one experiment accumulating 16 transients . The solvent resonance was depleted by excitation sculpting using a 180\u00b0 sinc pulse (duration\u00a0=\u00a02\u00a0ms) The NMR analyses employed a Bruker Avance III 700\u00a0MHz spectrometer with an inverse TCI cryoprobe optimised for +\u2010coupled) assay for quantification of the formaldehyde reaction byproduct, as previously described m HEPES, pH 7.5, 0.01% Tween\u201020, with addition of Fe(II) ammonium sulfate (10\u00a0\u03bcm), sodium ascorbate (100\u00a0\u03bcm), 2\u2010oxoglutarate (200\u00a0\u03bcm), NAD+ (500\u00a0\u03bcm), KDM4 enzymes and FDH enzyme in a 30\u2010\u03bcL volume in black 384\u2010well plates. Specific activities were measured using a 100\u2010\u03bcm peptide. For Michaelis\u2013Menten experiments, peptide concentrations were varied. Reactions were monitored using a PHERAstar FS plate reader with 355\u00a0nm excitation and 460\u00a0nm emission. Kinetic parameters were calculated from the reaction rate during the initial linear phase of formaldehyde production, which were used to calculate specific activities or fitted to Michaelis\u2013Menten equations using graphpad prism .Kinetic parameters for KDM4 enzymes were determined by use of an FDH /NAD1\u2013359.Ni(II).H1.4(18\u201332)K26me3.NOG were obtained in sitting drops grown at 4\u00a0\u00b0C with a ratio of 1\u00a0:\u00a02 sample to well solution [0.1\u00a0m MIB buffer ] \u22121 KDM4A, 5\u00a0mm H1.4(18\u201332)K26me3, 5\u00a0mm NOG and 4\u00a0mm NiCl2. Crystals were cryoprotected with 25% glycerol then flash\u2010frozen in liquid N2. Data were collected using a single crystal at 100\u00a0K at the Diamond I04\u20101 MX beam line and were processed with HKL2000 http://www.rcsb.org/pdb/search/structidSearch.do?structureId=2OX0) and was refined by alternative cycles of CNS http://www.rcsb.org/pdb/search/structidSearch.do?structureId=6H8P.Co\u2010crystals of KDM4A).H1.418\u2013K26me3.NO2N\u2010TPVKKKARKSAGAAK\u2010CONH2), but containing either a N\u03b5\u2010tri\u2010, di\u2010 or monomethyllysine at K26 were synthesised and fragment peptides Table\u00a0 37. Thresed Fig.\u00a0. Activitsed Fig.\u00a0. After iM4D Fig.\u00a0 15, 43. M4D Fig.\u00a0, low levM4D Fig.\u00a0.Given the activity observed with KDM4E and KDM7A, we decided to investigate the intra\u2010subfamily conservation of this demethylation activity. The demethylation activity of the other subfamily members (KDM4A\u2010D and KDM7B) was therefore screened. For all KDM4 subfamily members, we observed H1.4K26 demethylation activity similar to that already observed with KDM4E Table\u00a0; howeverN\u03b5\u2010Trimethylated H3K4 binds to the plant homeobox domain of KDM7B, targeting the N\u03b5\u2010dimethyllysine at H3K9 to the catalytic domain, and thus promoting demethylation di\u2010N\u03b5\u2010methyllysine at K26 and tri\u2010N\u03b5\u2010methyllysine at K21 K21me3K26me2) and the H1.4 peptides methylated only at K26 .K26 Fig.\u00a0, but no 1H NMR time course analyses with KDM4A and the trimethylated H1.4K26me3 peptide confirmed time\u2010dependent demethylation of the Kme3 residue were determined for comparison \u2010coupled demethylation assay, which monitors the formation of NADH during FDH\u2010catalysed oxidation of formaldehyde son Figs\u00a0D, S14. T+ and FDH were analysed over time and kinetic parameters were determined by recording the initial reaction rates of NADH production at varying peptide concentration. Similar experiments were carried out with H3K9me3 and H3K36me3 substrate peptides and the obtained values compared (Table\u00a0KM (32.3\u00a0\u00b1\u00a06.4\u00a0\u03bcm and 25.5\u00a0\u00b1\u00a04.9\u00a0\u03bcm respectively) and kcat (0.32\u00a0\u00b1\u00a00.049 s\u22121 and 0.31\u00a0\u00b1\u00a00.045 s\u22121 respectively) values. Demethylation\u00a0of the H3K36me3 peptide was the least efficient, with\u00a0higher KM (66.8\u00a0\u00b1\u00a05.4\u00a0\u03bcm) and lower kcat (0.12\u00a0\u00b1 0.004 s\u22121) values than with the other two peptides.Michaelis\u2013Menten kinetics were then conducted with KDM4A. Samples containing KDM4A, 2OG, ascorbate, H1.4K26me3 peptide, NADN\u2010oxalylglycine:H1.4K26me3 peptide complex was solved to a resolution of 1.98 \u00c5 (Note: Ni(II) was used for crystallisation in place of the catalytically relevant Fe(II) due to its oxidative stability; N\u2010oxalylglycine (NOG) is a 2OG mimetic inhibitor) Crystallographic analyses were undertaken to investigate the binding mode of trimethylated H1.4K26 in the KDM4A active site. An X\u2010ray crystal structure of a KDM4A:Ni(II):http://www.rcsb.org/pdb/search/structidSearch.do?structureId=2OQ6http://www.rcsb.org/pdb/search/structidSearch.do?structureId=2P5B\u2010 to C\u2010 directionality through the active site. The two residues N\u2010terminal to the trimethyllysine residue adopt a similar conformation/orientation in all three structures, with backbone hydrogen bonds between KDM4A E169 and the peptide residues at the \u22121 and \u22122 positions in its sequence. Thus, in the K36 peptide, the backbone carbonyl of the H3K36 to K37 amide bond is \u2018flipped\u2019 ~180\u00b0 compared to the analogous amide in the other two substrates :NOG:H3K9me3 peptide complex than either H3K9me3 or H1.4K26me3, binds differently at the KDM4A active site N\u2010methylated substrate lysine for H3K9/H1.4K26 versus H3K36 K26me3 crystal structure.Fig.\u00a0S2. KDM4E catalyses lysine demethylation at H1.4K26.Fig.\u00a0S3. KDM2A does not catalyse lysine demethylation at H1.4K26 under the tested conditions.Fig.\u00a0S4. KDM3A does not catalyse lysine demethylation at H1.4K26 under the tested conditions.Fig.\u00a0S5. KDM5C does not catalyse lysine demethylation at H1.4K26 under the tested conditions.Fig.\u00a0S6. KDM6B does not catalyse lysine demethylation at H1.4K26 under the tested conditions.Fig.\u00a0S7. KDM7A catalyses lysine demethylation at H1.4K26.Fig.\u00a0S8. KDM4A catalyses lysine demethylation at H1.4K26.Fig.\u00a0S9. KDM4B catalyses lysine demethylation at H1.4K26.Fig.\u00a0S10. KDM4C catalyses lysine demethylation at H1.4K26.Fig.\u00a0S11. KDM4D catalyses lysine demethylation at H1.4K26.Fig.\u00a0S12. PHF8/KDM7B only catalyses lysine demethylation at H1.4K26 at high concentration.Fig.\u00a0S13. Analysis of KDM4A demethylation by 1H NMR.Fig.\u00a0S14. Specific activity determination for KDM4 enzymes.Table\u00a0S1. Peptide sequences used in this study.Table\u00a0S2. Crystallographic data processing and refinement statistics.Click here for additional data file."} +{"text": "Brmind\u22efODio, C\u2014HBrmind\u22efOEthy, C\u2014HChlethy\u22efODio and C\u2014HChlethy\u22efOChlethy hydrogen bonds link the mol\u00adecules into a three-dimensional structure, enclosing The title compound consists of the 5-bromo\u00adindoline-2,3-dione unit linked by a 1-{2-[2-(2 chloro\u00adeth\u00adoxy)eth\u00adoxy]eth\u00adyl} moiety. In the crystal, inter\u00admolecular C\u2014H 14H15BrClNO4, consists of a 5-bromo\u00adindoline-2,3-dione unit linked to a 1-{2-[2-(2-chloro\u00adeth\u00adoxy)eth\u00adoxy]eth\u00adyl} moiety. In the crystal, a series of C\u2014H\u22efO hydrogen bonds link the molecules to form a supramolecular three-dimensional structure, enclosing R22(8), R22(12), R22(18) and R22(22) ring motifs. \u03c0\u2013\u03c0 contacts between the five-membered dione rings may further stabilize the structure, with a centroid\u2013centroid distance of 3.899\u2005(2)\u2005\u00c5. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (28.1%), H\u22efO/O\u22efH (23.5%), H\u22efBr/Br\u22efH (13.8%), H\u22efCl/Cl\u22efH (13.0%) and H\u22efC/C\u22efH (10.2%) inter\u00adactions. Hydrogen bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Density functional theory (DFT) optimized structures at the B3LYP/6-311G level are compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2014LUMO behaviour was elucidated to determine the energy gap. The chloro\u00adeth\u00adoxy\u00adethoxyethyl side chain atoms are disordered over two sets of sites with an occupancy ratio of 0.665\u2005(8):0.335\u2005(6).The title compound, C They are very inter\u00adesting chemical compounds because of their potential applications in different fields. The most common heterocycles contain nitro\u00adgen and oxygen and B (N1/C1/C6\u2013C8), are oriented at a dihedral angle of A/B = 2.78\u2005(6)\u00b0. Atoms Br1, O1 and C9 are at distances of 0.0415\u2005(4), 0.0464\u2005(8) and \u22120.0244\u2005(7)\u2005\u00c5, respectively, from the best plane of the bromo\u00adindoline unit. The 1-{2-[2-(2-chloro\u00adeth\u00adoxy)eth\u00adoxy]eth\u00adyl} moiety is oriented with respect to the bromo\u00adindoline unit by 77.7\u2005(2)\u00b0, as defined by the C10\u2014C9\u2014N1\u2014C1 torsion angle.The title compound, (I)Brmind\u22efODio, C\u2014HBrmind\u22efOEthy, C\u2014HChlethy\u22efODio and C\u2014HChlethy\u22efOChlethy hydrogen bonds (Table\u00a01Cg1\u2014Cg1i , may further stabilize the structure, with a centroid\u2013centroid distance of 3.899\u2005(2)\u2005\u00c5. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (28.1%), H\u22efO/O\u22efH (23.5%), H\u22efBr/Br\u22efH (13.8%), H\u22efCl/Cl\u22efH (13.0%) and H\u22efC/C\u22efH (10.2%) inter\u00adactions. Hydrogen bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing.In the crystal, inter\u00admolecular C\u2014Hs Table\u00a01 link thefs Fig.\u00a02. \u03c0\u2013\u03c0 conCrystalExplorer17.5 and those delineated into H\u22efH, H\u22efO/O\u22efH, H\u22efBr/Br\u22efH, H\u22efCl/Cl\u22efH, H\u22efC/C\u22efH, O\u22efC/C\u22efO, C\u22efC and O\u22efCl/Cl\u22efO contacts \u2013(i), respectively, together with their relative contributions to the Hirshfeld surface. The most important inter\u00adaction is H\u22efH, contributing 28.1% to the overall crystal packing, which is reflected in Fig.\u00a06b) as widely scattered points of high density due to the large hydrogen content of the mol\u00adecule with the tip at de = di \u223c1.08\u2005\u00c5, due to the short inter\u00adatomic H\u22efH contacts (Table\u00a02c), with a 23.5% contribution to the HS, arises from the H\u22efO/O\u22efH contacts and H\u22efCl/Cl\u22efH contacts, with 13.8 and 13.0% contributions to the HS, have nearly symmetrical distributions of points with the edges at de + di = 2.92 (for thin edge) and 3.20\u2005\u00c5 (for thick edge) and de + di = 2.78\u2005\u00c5, respectively, arising from the H\u22efBr/Br\u22efH and H\u22efCl/Cl\u22efH contacts (Table\u00a02f), a 10.2% contribution to the HS, arises from the H\u22efC/C\u22efH contacts , with a 4.0% contribution to the HS, arises from the O\u22efC/C\u22efO contacts , with a 2.6% contribution to the HS, have a nearly arrow-shaped distribution of points arising from the C\u22efC contacts contacts, with a 1.1% contribution to the HS, have nearly symmetrical distributions of points with the edge at de + di = 3.50\u2005\u00c5.In order to visualize the inter\u00admolecular inter\u00adactions in the crystal of the title compound, a Hirshfeld surface (HS) analysis \u2013(e), respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing using standard B3LYP functional and 6-311G basis-set calculations of the mol\u00adecule was about 6.5402\u2005eV, and the frontier mol\u00adecular orbital energies, i.e.EHOMO and ELUMO, were \u22127.4517 and \u22120.9115\u2005eV, respectively.The optimized structure of the title compound, (I)et al., 2010et al., 2010H-indole-2,3-dione and di\u00admethyl\u00adformamide in potassium carbonate and tetra-n-butyl\u00adammonium bromide . The mixture was stirred at 353\u2005K for 48\u2005h. The reaction was controlled by CCM. The solution was filtered and the DMF was removed under vacuum. The product obtained was separated by chromatography on a column of silica gel with hexa\u00adne\u2013ethyl acetate (4:1 v/v) as eluent. The isolated solid was recrystallized from ethanol to afford red crystals .1,2-Bis(2-chloro\u00adeth\u00adoxy)ethane was added dropwise to a solution of 5-bromo-1Uiso(H) = 1.2Ueq(C). During the refinement process, the disordered chloro\u00adeth\u00adoxy\u00adethoxyethyl side-chain atoms were refined with a major\u2013minor occupancy ratio of 0.665\u2005(8):0.335\u2005(6).The experimental details, including the crystal data, data collection and refinement, are summarized in Table\u00a0310.1107/S2056989019011617/lh5913sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019011617/lh5913Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019011617/lh5913Isup3.cdxSupporting information file. DOI: 1948316CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structures of two first-row transition metal (Fe and Co) pyridine\u2013sulfate complexes are presented. The compounds demonstrate infinite chains of metal pyridine units connected by bridging sulfate anions. catena-poly[[tetra\u00adkis\u00ad(pyridine-\u03baN)iron(II)]-\u03bc-sulfato-\u03ba2O:O\u2032], [Fe(SO4)(C5H5N)4]n, (1), and catena-poly[[tetra\u00adkis\u00ad(pyridine-\u03baN)cobalt(II)]-\u03bc-sulfato-\u03ba2O:O\u2032-[tetra\u00adkis\u00ad(pyridine-\u03baN)cobalt(II)]-\u03bc-sulfato-\u03ba3O,O\u2032:O\u2032\u2032-[tris\u00ad(pyridine-\u03baN)cobalt(II)]-\u03bc-sulfato-\u03ba2O:O\u2032], [Co3(SO4)3(C5H5N)11]n, (2), are reported. The iron compound (1) displays a polymeric structure, with infinite chains of FeII atoms adopting octa\u00adhedral N4O2 coordination environments that involve four pyridine ligands and two bridging sulfate ligands. The cobalt compound (2) displays a polymeric structure, with infinite chains of CoII atoms. Two of the three Co centers have an octa\u00adhedral N4O2 coordination environment that involves four pyridine ligands and two bridging sulfate ligands. The third Co center has an octa\u00adhedral N3O3 coordination environment that involves three pyridine ligands, and two bridging sulfate ligands with one sulfate chelating the cobalt atom.The solid-state structures of two metal\u2013pyridine\u2013sulfate compounds, namely The first reports of a pyridine\u2013sulfato\u2013metal complex were in the late 19th century and the cobalt\u2013pyridine\u2013sulfate (2) complexes.Against this backdrop, our lab has recently begun to study the solid-state structures of transition-metal pyridine complexes. We have recently reported the structures of nickel, copper and zinc pyridine sulfates, which showed varying coordination geometries consistent with those predicted by crystal field theory (Roy 1), the asymmetric unit consists of two pyridine mol\u00adecules and one half of a sulfate anion coordinated to an iron atom sitting on an inversion center . When grown out, the iron displays an octa\u00adhedral coordination environment . There is a square-planar tetra\u00adpyridine iron unit, with FeN4 planarity enforced by the inversion. The octa\u00adhedral coordination is completed by two sulfate ions that bind trans to each other. The cis N\u2014Fe\u2014N angles have values of 86.44\u2005(4) and 93.56\u2005(4)\u00b0 and the cis O\u2014Fe\u2014N angles have values ranging from 88.12\u2005(4) to 91.88\u2005(4)\u00b0. The pyridine rings are rotated from the FeN4 plane by dihedral angles of 44.03\u2005(1) and 78.20\u2005(1)\u00b0. The 78.20\u2005(1)\u00b0 angle is constrained by two C\u2014H\u22efO inter\u00adactions with the trans sulfates , the asymmetric unit consists of three cobalt atoms, eleven coordinated pyridine mol\u00adecules, and three sulfate anions . There are three crystallographically independent cobalt atoms, with Co1 and Co2 displaying octa\u00adhedral N4O2 coordination environments, and Co3 showing an octa\u00adhedral N3O3 coordination environment .In the pink crystals of to 93.21\u2005(9)\u00b0, and the O\u2014Co\u2014O angle is 174.62\u2005(9)\u00b0. The four pyridine rings are rotated from the CoN4 plane by dihedral angles of 37.51\u2005(1), 45.21\u2005(1), 56.40\u2005(1) and 56.92\u2005(1)\u00b0. Two of the rings form one C\u2014H\u22efO inter\u00adaction each with the sulfate oxygen atoms to 93.19\u2005(9)\u00b0, and the O\u2014Co\u2014O angle is 175.16\u2005(9)\u00b0. The four pyridine rings are rotated from the CoN4 plane by dihedral angles of 55.37\u2005(1), 65.88\u2005(1), 67.08\u2005(1) and 68.07\u2005(1)\u00b0. Two of the rings are involved in two C\u2014H\u22efO inter\u00adactions each with the sulfate oxygen atoms that form a CoO3 plane with a maximum deviation from planarity of 0.029\u2005\u00c5. The meridional CoN3 and CoO3 planes are rotated relative to one another by an angle of 88.93\u2005(1)\u00b0. The cis N\u2014Co\u2014N angles have values of 86.76\u2005(10) and 87.52\u2005(9)\u00b0. The chelating sulfate exhibits an O\u2014Co\u2014O bite angle of 65.36\u2005(7)\u00b0 and another cis O\u2014Co\u2014O angle of 88.63\u2005(8)\u00b0. The three pyridine rings are rotated from the CoN3 plane by dihedral angles of 31.855\u2005(2), 44.111\u2005(3) and 82.863\u2005(4)\u00b0. The 82.863\u2005(4)\u00b0 angle is constrained by two C\u2014H\u22efO inter\u00adactions with sulfate oxygen atoms , the FeII atoms are linked together into infinite chains along the [100] direction through the sulfate ligands via O\u2014S\u2014O bridges . Between each successive tetra\u00adpyridine iron unit are found parallel slipped \u03c0\u2013\u03c0 inter\u00adactions .In compound , the CoII atoms linked together into infinite chains along the [001] direction through the sulfate ligands . No \u03c0\u2013\u03c0 inter\u00adactions are observed in this crystal. There are two C\u2014H\u22efO inter\u00adactions between chains that connect the chains in three dimensions and steric effects could play significant roles in determining the observed coordination environments.In a report earlier this year, we presented the structures of the metal\u2013pyridine\u2013sulfates of nickel, copper and zinc. It was of note that these three structures exhibited different coordination geometries, consistent with the crystal field stabilization energies (CFSE) associated with their Approximately 25\u2005mg of each metal sulfate were dissolved in pyridine in a 20\u2005mL vial under an atmosphere of di\u00adnitro\u00adgen. In the cobalt case, 0.1\u2005mL of water was also added. The vials were heated to 353\u2005K for 24\u201348\u2005h, after which single crystals suitable for X-ray diffraction studies were isolated.SHELXL) by full-matrix least squares on F2. Hydrogen atoms were placed in calculated positions and then refined with a riding model with C\u2014H bond lengths of 0.95\u2005\u00c5 and with isotropic displacement parameters set to 1.20 Ueq of the parent C atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018007557/sj5556sup1.cifCrystal structure: contains datablock(s) 1, 2. DOI: 10.1107/S2056989018007557/sj55561sup4.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989018007557/sj55562sup5.hklStructure factors: contains datablock(s) 2. DOI: 1844143, 1844142CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title structure has a square-planar coordination sphere around the copper(II) ion. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 hydrogen bonds and very weak \u03c0-stacking inter\u00adactions, forming a three-dimensional supra\u00admolecular architecture. 18H12F6N2O4)]\u00b70.5C6H6O2, the CuII ion has a square-planar coordination geometry, being ligated by two N and two O atoms of the tetra\u00addentate open-chain Schiff base ligand 6,6\u2032-{-bis\u00ad(methanylyl\u00adidene)}bis\u00ad[2-(tri\u00adfluoro\u00admeth\u00adoxy)phenol]. The crystal packing is stabilized by intra\u00admolecular O\u2014H\u22efO and inter\u00admolecular C\u2014H\u22efF, C\u2014H\u22efO and C\u2014H\u22ef\u03c0 hydrogen bonds. In addition, weak \u03c0\u2013\u03c0 inter\u00adactions form a three-dimensional structure. Hirshfeld surface analysis and two-dimensional fingerprint plots were performed and created to analyze the inter\u00admolecular inter\u00adactions present in the crystal, indicating that the most important contributions for the crystal packing are from F\u22efH/H\u22efF (25.7%), H\u22efH (23.5%) and C\u22efH/H\u22efC (12.6%) inter\u00adactions.In the title com\u00adplex, [Cu(C In this study, we describe the crystal structure and Hirshfeld surface analysis of the title com\u00adpound, as determined by X-ray crystallographic analysis.In this study, a salen-type Schiff base has been synthesized from 2-hy\u00addroxy-5-(tri\u00adfluoro\u00admeth\u00adoxy)benzaldehyde with ethyl\u00adenedi\u00adamine by a condensation reaction. The synthesized Schiff base was used as an II ion chelated by a doubly deprotonated tetra\u00addentate Schiff base ligand and a hydrogen-bonded mol\u00adecule of hydro\u00adquinone. The Cu1 ion is coordinated by two imine N atoms (N6 and N7) and two phenoxo O atoms (O2 and O3) of the tetra\u00addentate Schiff base ligand 6,6\u2032-{-bis\u00ad(methanylyl\u00adidene)}bis\u00ad[2-(tri\u00adfluoro\u00admeth\u00adoxy)phenol] (L1). The hydro\u00adquinone mol\u00adecule is located on an inversion centre and is linked to neighbouring com\u00adplex cations via O\u2014H\u22efO hydrogen bonds. The bond lengths Cu1\u2014O2 and Cu1\u2014O3 and Cu1\u2014N1 and Cu1\u2014N2 are close to the values observed for related copper(II) com\u00adplexes reported in the literature hydrogen bonds (Table\u00a02Cg2\u22efCg3 distance is 3.507\u2005(2)\u2005\u00c5, where Cg2 and Cg3 are the centroids of the Cu1/O2/C5/C6/C8/N2 and Cu1/O3/C17/C12/C11/N1 rings, respectively.The crystal packing of the title com\u00adpound is stabilized by inter\u00admolecular C\u2014H\u22efO, C\u2014H\u22efF and C\u2014H\u22efs Table\u00a02. In addis Table\u00a02. The Cg2CrystalExplorer17 . The blue, white and red colour conventions used for the dnorm-mapped Hirshfeld surfaces recognize the inter\u00adatomic contacts as longer, at van der Waals separations and short inter\u00adatomic contacts, respectively.The Hirshfeld surface analysis pair with the full fingerprint plot outlined in gray. Fig.\u00a05a) shows the two-dimensional fingerprint plot of the sum of the contacts contributing to the Hirshfeld surface represented in normal mode. The most significant contribution to the Hirshfeld surface is from F\u22efH/H\u22efF contacts (25.7%) . Here, H\u22efH interactions are only the second most significant contribution to the total Hirshfeld surface (23.5%). In addition, C\u22efH/H\u22efC and O\u22efH/H\u22efO contacts contribute 12.6 and 11.2% to the Hirshfeld surface, respectively.A fingerprint plot delineated into specific inter\u00adatomic contacts contains information related to specific inter\u00admolecular inter\u00adactions. The blue colour refers to the frequency of occurrence of the (%) Fig.\u00a05b. Here,et al., 20163 and 1.929\u2005(5)\u2005\u00c5] fall within these limits. While the Cu1\u2014O3 and C1\u2014O2 bond length are within and close to these limits, respectively, the Cu1\u2014O2 bond length is outside these limits, with a shorter value of 1.883\u2005(4)\u2005\u00c5.A search of the Cambridge Structural Database -bis\u00ad(methan\u00adylyl\u00adidene)}bis\u00ad[4-(tri\u00adfluoro\u00admeth\u00adoxy)phenol] (H2L1) was syn\u00adthesized by condensation of 2-hy\u00addroxy-5-(tri\u00ad\u00adfluoro\u00admeth\u00adoxy)\u00adbenzaldehyde (0.0095\u2005mmol) and 1,2-ethane\u00addiamine (0.0095\u2005mmol) in ethanol under reflux for about 18\u2005h. The yellow product was washed with ether and dried at room temperature. 0.0080\u2005mmol\u2005H2L1 was dissolved in 20\u2005ml ethanol and 0.0080\u2005mmol Cu(CH3COO)2\u00b7H2O was dissolved in 20\u2005ml ethanol. The metal solution was added dropwise to the Schiff base solution and the resulting solution refluxed for about 6\u2005h. The product (CuL1) was washed with toluene and crystallized from ethanol at room temperature. 2,2\u2032-{-bis\u00ad(methanylyl\u00adidene)}bis\u00ad[4-(tri\u00adfluoro\u00admeth\u00adoxy)phenol]copper(II) hydro\u00adquinone hemisolvate was obtained even after 0.0040\u2005mmol hydro\u00adquinone was added to 0.0040\u2005mmol CuL1 in 20\u2005ml ethanol and refluxed for about 6\u2005h. A purple crystal suitable for X-ray diffraction analysis was obtained from the reaction Fig.\u00a06.Uiso(H)\u00a0= 1.2Ueq(C) for methyl\u00adene, C\u2014H = 0.93\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for aromatic, C\u2014H = 0.93\u2005\u00c5 and Uiso(H)\u00a0= 1.2Ueq(C) for methine, and O\u2014H = 0.82\u2005\u00c5 and Uiso(H) = 1.5Ueq(O) for hy\u00addroxy H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019014294/lh5932sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019014294/lh5932Isup2.hklStructure factors: contains datablock(s) I. DOI: 1900670, 1900670CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title Schiff base compound was obtained from a condensation reaction of 4-chloro-3-hy\u00addroxy\u00adbenzaldehyde and 2,4-di\u00adnitro\u00adphenyl\u00adhydrazine. The mol\u00adecule is almost planar with the dihedral angle between the benzene rings being 3.70\u2005(17)\u00b0. 13H9ClN4O5\u00b70.5CH3CN, crystallizes as an aceto\u00adnitrile hemisolvate; the solvent mol\u00adecule being located on a twofold rotation axis. The mol\u00adecule is nearly planar, with a dihedral angle between the two benzene rings of 3.7\u2005(2)\u00b0. The configuration about the C=N bond is E, and there is an intra\u00admolecular N\u2014H\u22efOnitro hydrogen bond present forming an S(6) ring motif. In the crystal, mol\u00adecules are linked by O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds, forming layers lying parallel to (10The title Schiff base compound, C These compounds show biological activities including anti\u00adbacterial, anti\u00adfungal, anti\u00adcancer and herbicidal activities ring motif \u2005\u00c5, where Cg1 and Cg2 are the centroids of rings C1\u2013C6 and C8\u2013C13, respectively, \u03b1 = 3.70\u2005(17)\u00b0, \u03b2 = 27.9\u00b0, \u03b3 = 24.5\u00b0, inter\u00adplanar distances are 3.489\u2005(2) and 3.388\u2005(2)\u2005\u00c5, offset = 1.791\u2005\u00c5; symmetry code: (i) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01. There are no other significant inter\u00admolecular contacts present in the crystal.In the crystal, mol\u00adecules are linked by O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds Table\u00a01, formingGAUSSIAN09 level . 18 of these structures involve a halide substituent and 23 involve a hydroxyl substituent. Only one compound involves both a halide and an hydroxyl substituent and closely resembles the title compound, viz. 4-chloro-2-{[hydra\u00adzono]meth\u00adyl}phenol ring motif. In fact, in all 71 structures (see supporting information) there is an intra\u00admolecular N\u2014H\u22efOnitro hydrogen bond present forming an S(6) ring motif, and in the majority of the compounds the two benzene rings are almost coplanar with the dihedral angle varying between ca 0 to 8\u00b0, with a few exceptions.A search of the Cambridge Structural Database in ethanol (15\u2005ml) and 2,4-di\u00adnitro\u00adphenyl\u00adhydrazine in ethanol (15\u2005ml). The reaction mixture was stirred for 5\u2005h under reflux. Orange plate-like crystals of the title compound were obtained by slow evaporation of a solution in ethanol .Uiso(H) = 1.5Ueq and 1.2Ueq for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901900642X/su5496sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S205698901900642X/su5496Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S205698901900642X/su5496sup3.pdfCSD search S1. DOI: Click here for additional data file.10.1107/S205698901900642X/su5496Isup4.cmlSupporting information file. DOI: 1912273CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title chalcone derivative is almost planar, with a dihedral angle of 7.0\u2005(2)\u00b0 between the 3,5-di\u00adchloro-2-hy\u00addroxy\u00adphenyl and 5-methyl\u00adfuran rings. 14H10Cl2O3, is almost planar, with a dihedral angle of 7.0\u2005(2) \u00b0 between the 3,5-di\u00adchloro-2-hy\u00addroxy\u00adphenyl and 5-methyl\u00adfuran rings. There is an intra\u00admolecular O\u2014H\u22efO hydrogen bond present forming an S(6) ring motif. In the crystal, mol\u00adecules are linked by bifurcated C\u2014H/H\u22efO hydrogen bonds, enclosing an R12(6) ring motif, forming a 21 helix propagating along the b-axis direction. The inter\u00admolecular inter\u00adactions were qu\u00adanti\u00adfied using Hirshfeld surface analysis.The title chalcone derivative, C They belong to the flavonoid family, which are basically found in fruits and vegetables -1--3-(5-methyl\u00adfuran-2-yl)prop-2-en-1-one.Chalcone derivatives are an important class of organic compounds comprising two aromatic rings connected via an unsaturated \u03b1,\u03b2 carbonyl system as shown in Fig.\u00a01S(6) ring motif (Table\u00a01anti-periplanar conformation described by the torsion angles C11\u2014C12\u2014C13\u2014Cl19 = \u2212179.1\u2005(3)\u00b0 and C13\u2014C14\u2014C15\u2014Cl18 = \u2212178.6\u2005(4)\u00b0, while methyl group at C2 of the furan ring is in a +anti-periplanar conformation [C5\u2014O1\u2014C2\u2014C6 = 178.3\u2005(4)\u00b0]. The bond lengths and angles in the title compound are similar to those observed for 3-(furan-2-yl)-1-(2-hy\u00addroxy\u00adphen\u00adyl)prop-2-en-1-one \u2005\u00c5, propagating along the b-axis direction < 1.8\u2005\u00c5 (di is the distance of a point on the Hirshfeld surface to the nearest nucleus inside the surface while de is the distance of the nearest nucleus outside the surface). The H\u22efH contacts, with a contribution of 25.7%, are shown as blue dots spread in the middle region 1.18\u2005\u00c5 < (de + di) < 1.62\u2005\u00c5. The two sharp spikes observed at 1.04\u2005\u00c5 < (de + di) < 1.39\u2005\u00c5 are due to the presence of a pair of O\u22efH contacts making a 15.2% contribution. A pair of C\u22efH contacts are observed as characteristic wings in the region of 1.18\u2005\u00c5 < (de + di) < 1.6\u2005\u00c5 (13.0% contribution). C\u22efC, C\u22efCl and O\u22efC contacts make contributions of 7.9%, 5.2% and 3.8%, respectively.The two-dimensional fingerprint Fig.\u00a06 plots weet al., 2016et al., 2015et al., 2014et al., 2014S(6) ring motifs. The mol\u00adecules are all relatively planar with the dihedral angle between the furan and 2-hy\u00addroxy\u00adphenyl rings varying from ca 8.35\u00b0 in BOGVID, 0.20\u00b0 in KUDMON, and 10.90 and 2.56\u00b0 in the two independent mol\u00adecules of POJBAT. The only exception is POHZUJ, which possesses twofold rotation symmetry and has two [3-(2-fur\u00adyl)prop-2-en-1-one] units meta to each other; here the dihedral angle is ca 19.87\u00b0.A search of the Cambridge Structural Database -2-hy\u00addroxy\u00adethanone (5\u2005mmol) was dissolved in methanol (15\u2005ml) and was stirred with 5\u2005ml of sodium hydroxide solution for 30\u2005min at room temperature. To this mixture, 5-methyl\u00adfuran-2-carbaldehyde (5\u2005mmol) was added over 30\u2005min with stirring. Stirring at room temperature was then continued for 32\u2005h. On completion of the reaction, monitored by TLC, the mixture was quenched in ice\u2013water and acidified with dilute hydro\u00adchloric acid. The separated precipitate of the title compound was filtered off and recrystallized from methanol solution giving colourless block-like crystals.Uiso(H) = 1.2Ueq(C) for aromatic H atoms and C\u2014H = 0.96\u2005\u00c5 with Uiso(H) = 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018012173/qm2127sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018012173/qm2127Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018012173/qm2127Isup3.cmlSupporting information file. DOI: 1852049CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "WC)], reverse Watson-Crick [A\u00b7T(wrWC)], Hoogsteen [A\u00b7T(wH)] and reverse Hoogsteen [A\u00b7T(wrH)] \u2013 act as intermediates of the intrapair mutagenic tautomerization of the T nucleobase owing to the novel tautomerisation pathways: A\u00b7T(wWC)\u2194A\u00b7T*(w\u22a5WC); A\u00b7T(wrWC)\u2194A\u00b7\u22a5rWC); A\u00b7T(wH)\u2194A\u00b7T*(w\u22a5H); A\u00b7T(wrH)\u2194A\u00b7\u22a5rH). All of them occur via the transition states as tight ion pairs \u00b7 with quasi-orthogonal geometry, which are stabilized by the participation of the strong (A)N6+H\u00b7\u00b7\u00b7O4\u2212/O2\u2212(T) and (A)N6+H\u00b7\u00b7\u00b7N3\u2212(T) H-bonds. Established tautomerizations proceed through a two-step mechanism of the protons moving in the opposite directions along the intermolecular H-bonds. Initially, proton moves from the N3H imino group of T to the N6H2 amino group of A and then subsequently from the protonated N6+H3 amino group of A to the O4/O2 oxygen atom of T, leading to the products \u2013 A\u00b7T*(w\u22a5WC), A\u00b7\u22a5rWC), A\u00b7T*(w\u22a5H), and A\u00b7\u22a5rH), which are substantially non-planar, conformationally-labile complexes. These mispairs are stabilized by the participation of the (A)N6H/N6H'\u00b7\u00b7\u00b7N3(T) and (T)O2H/O4H\u00b7\u00b7\u00b7N6(A) H-bonds, for which the pyramidalized amino group of A is their donor and acceptor. The Gibbs free energy of activation of these mutagenic tautomerizations lies in the range of 27.8\u201329.8 kcal\u00b7mol\u22121 at T = 298.15 K in the continuum with \u03b5 = 1.In this paper using quantum-mechanical (QM) calculations in combination with Bader's quantum theory of \u201cAtoms in Molecules\u201d (QTAIM) in the continuum with \u03b5 = 1, we have theoretically demonstrated for the first time that revealed recently highly-energetic conformers of the classical A\u00b7T DNA base pairs \u2013 Watson-Crick [A\u00b7T(w Protonated amino group N6+H3 of A for these TSs acts simultaneously as donor and acceptor of the H-bonding and has such spatial orientation, that its N6+H/N6+H\u2032 bond, which is not involved in the H-bonding with T, lies in the plane of the purine ring R, L (10.44), A\u00b7T*(w\u22a5H)R, L (14.69), A\u00b7\u22a5rWC)R, L (9.20) and A\u00b7\u22a5rH)R, L (13.75) kcal\u00b7mol\u22121 complexes (C1 symmetry), which are the products of these mutagenic tautomerizations, are stabilized by the two anti-parallel (T)O4H/O2H\u00b7\u00b7\u00b7\u00a0N6(A) (~5.5) and (A)N6H/N6H\u2032\u00b7\u00b7\u00b7\u00a0N3(T) H-bonds R, L 0.44, A\u00b7T\u22a5WC)R, L 0.44, A\u00b7Tcys/trans mutual orientation of the N1H and N9H glycosydic bonds of the bases. At the mutagenic tautomeric transformations of the DNA bases some R/L structures transfer into the other L/R structures and vice versa and \u22121) with low values of the imaginary frequencies provided in the brackets. At this, one-single intermolecular (T)O4H/O2H\u00b7\u00b7\u00b7\u00a0N6(A) H-bond between the O4H/O2H hydroxyl groups of T*/1s. In the case of TS2s, when T hangs over A, the (T)O4H/O2H\u00b7\u00b7\u00b7\u00a0N6(A) H-bond coexists together with attractive van der Waals contacts with significantly increased ellipticity \u2013 N3\u00b7\u00b7\u00b7\u00a0C6 and O2\u00b7\u00b7\u00b7\u00a0C4 in the case of 1s are the most energetically favorable (1.86 and 1.92) in comparison with the TS2s (Table Two of these tautomerization reactions are controlled by the TSs \u2013 vice versa. In these cases TSs \u2013 \u22121) and \u22121) are characterized by the considerably higher values of imaginary frequencies and stabilized by two antiparallel (T)O4/O2H\u00b7\u00b7\u00b7\u00a0N6(A) and (A)N6H/N6H\u2032\u00b7\u00b7\u00b7\u00a0N3(T) H-bonds, the first of which is significantly stronger, then the other one contribute only a part of the electron energy of the monomer interactions R,L\u2194 A\u00b7T*(w\u22a5H)R,L and A\u00b7T*O2(*O2(w\u22a5rH)R,L, tautomerizations indicating the close structural relationship between tautomerization the classical A\u00b7T(WC) and A\u00b7T(H) DNA base pairs, on the one hand, and A\u00b7T(rWC) and A\u00b7T(rH), on the other hand ], reverse Watson-Crick [A\u00b7T(wrWC)], Hoogsteen [A\u00b7T(wH)] and reverse Hoogsteen [A\u00b7T(wrH)], which have been analyzed in details in our previous paper \u2194A\u00b7T*(w\u22a5WC), A\u00b7T(wrWC)\u2194A\u00b7\u22a5rWC), A\u00b7T(wH)\u2194A\u00b7T*(w\u22a5H), A\u00b7T(wrH)\u2194A\u00b7\u22a5rH) \u2013 proceed through the stepwise proton transfer via the TSs as tight A+\u00b7T\u2212 ion pairs, which Gibbs free energy of activation lies in the range of 27.79\u201329.83 kcal\u00b7mol\u22121 at T = 298.15 K, thus creating the substantially non-planar, conformationally-labile complexes \u2013 A\u00b7T*(w\u22a5WC), A\u00b7\u22a5rWC), A\u00b7T*(w\u22a5H) and A\u00b7\u22a5rH). Furthermore, formed complexes involving mutagenic T*/*(w\u22a5WC)\u2194A\u00b7T*(w\u22a5H) and A\u00b7\u22a5rWC)\u2194A\u00b7\u22a5rH).Here we have shed light on the revealed for the first time physico-chemical mechanism of the intrapair mutagenic tautomerization of the T DNA base within the novel highly-energetic conformers of the classical A\u00b7T DNA base pairs \u2013 Watson-Crick [A\u00b7T(wOB, study conception and design, acquisition of data, drafting of manuscript analysis and interpretation of data, performance of calculations, discussion of the obtained data, preparation of the numerical data for Tables, graphical materials for Figures and text of the manuscript. KT, preparation of the numerical data for Tables and graphical materials for Figures, preparation of the text of the manuscript. AD, analysis and preparation of the current literature survey, discussion of the strategy of the current investigation, analysis of the obtained numerical data, discussion of the obtained data, preparation of the numerical data for Tables, graphical materials for Figures and text of the manuscript. DH, study conception, critical revision of manuscript, proposition of the task of the investigation, discussion of the obtained data, preparation of the text of the manuscript. All authors were involved in the proofreading of the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The proposed approach permits the selection of hydrolases belonging to different families and active towards different substrates. Moreover, the ester group of the substrate used for the selection, at least partly, determined the specificity of the selected enzymes.A high\u2010throughput method (\u2265\u00a010 Enzyme\u2010based organic synthesis constantly needs innovative, diverse and robust biocatalysts construction of metagenomic libraries, and subsequent functional screening or by using tributyrin\u2010supplemented agar plates \u2010 and (S)\u2010enantiomers has been used as a substrate in the bond\u2010breaking reaction that generates a growth\u2010promoting energy source and a growth\u2010inhibiting compound and were omitted from the further analysis. The plasmid DNA from the remaining clones was isolated, and the fragments obtained after restriction digestion were analysed by sequencing to exclude the redundancy. Hence, 30 clones were chosen for further analysis.The metagenomic DNA isolated from different soil samples was partially digested with several restriction endonucleases, and the fragmented DNA was used for the construction of metagenomics libraries. In total, 19 libraries were tesBioinformatics analysis showed that the selected clones exhibiting esterase activity contained ORFs with medium 31%) to high (100%) sequence identity to proteins found in the databases (Table\u00a0% to highN\u2010acyltransferase superfamily (SSF55729) (1 hit) and one DUF998 family protein revealed that the enzyme 1315H may be a transmembrane protein without any predictable function.The phylogenetic analysis of the selected hydrolases showed that the enzymes represent very diverse groups of proteins Fig.\u00a0. Most ofein Fig.\u00a0. All ideein Fig.\u00a0 and wereein Fig.\u00a0. Two estThus, a functional selection of metagenomic libraries revealed enzymes from very diverse protein families, including several hits that, based on bioinformatics, could not be annotated as hydrolases active towards esters.E.\u00a0coli strain BL21 (DE3) was transformed with the recombinant plasmids and used for the expression of the recombinant proteins. In total, 27 recombinant proteins were purified by Ni\u2010NTA chromatography esters: acetate, butyrate, valerate, decanoate, palmitate and stearate. All hydrolases were active towards the short\u2010chain esters pNP\u2010acetate and pNP\u2010butyrate, most of them used pNP\u2010valerate, but only a few of them hydrolyzed pNP\u2010decanoate exhibited a very weak activity against this ester. Six enzymes MO101T, SVG3, GRU1, PLA1, 3T and 33T3 exhibited a strong preference for pNP\u2010acetate as the substrate forming the halos indicative of the hydrolysis were screened. Tb7_1T (MH423281) encoded the group \u03b2\u2010lactamases protein, and Tb10_7T (MH423280) was most similar to ABhydrolases. Neither of the two clones was capable of growing on compound 1 or 2 as a uridine source. Tb7\u20101T was not further analysed because of its insolubility. The purified protein Tb10_7 was not only active towards pNP esters and two esterases preferred S\u20101\u2010phenylethyl hexanoate and S\u20101\u2010phenylethyl benzoate respectively. No esterase was enantioselective towards R\u20101\u2010phenylethyl hexanoate or benzoate. Furthermore, to analyse the promiscuity of the selected esterases, several amides were tested as substrates. Consequently, three esterases were found to hydrolyze acetanilide, and 18 were active towards nitrocefin and the uridine auxotroph mutant \u0394pyrFEC of E.\u00a0coli allows for a functional selection of novel ester hydrolases from metagenomic libraries. Evidently, the selection method presented here is highly complementary to traditional approaches and is applicable for allowing the discovery of novel esterases with different structural and catalytic characteristics. Compared with the known methods, the proposed selection system has many advantages: (i) it is a HTS method that allows for a rapid (1\u20134\u00a0days) processing of large (meta)genome libraries with low number of false positives, ii) depending on the selection media used, it permits the functional selection of hydrolases belonging to different protein families, (iii) it is flexible since, by using different substrates, for example, esters of the fatty acids of different lengths or bulky carboxylic acids, it allows the identification of enzymes with desired properties, (iv) it would be suitable for the selection of regioselective esterases if the appropriate substrate was used, (v) it allows the identification of novel proteins that previously have not been known as the ester\u2010hydrolysing enzymes or cannot be screened using, for example, a tributyrin method, (vi) the appropriate uridine auxotrophs are available for a wide spectra of microbial hosts, including extremophiles and yeasts or can be easily constructed, for example, using 5\u2010fluororotic acid selection Assay Reagent and HisPur\u2122 Ni\u2010NTA spin column were purchased from Thermo Fisher Scientific, Rockford, IL. pET21a Vector was purchased from Novagen . Nutrient medium were purchased Roth, Germany. pNP\u2010acyl esters, tributyrin, \u03b2\u2010d\u2010glucose pentaacetate, \u03b2\u2010d\u2010galactose pentaacetate, and uridine were purchased from Sigma\u2010Aldrich. \u2018ZR Soil Microbe DNA MidiPrep\u2122\u2019 was purchased from Zymo Research, Freiburg, Germany. Nitrocefin was purchased from Oxoid, UK. 3\u2032\u2010O\u2010benzoyl\u20102\u2032\u2010deoxyuridine were purchased from Carbosynth, UK. 3\u2032\u2010O\u2010acetyl\u20102\u2032\u2010deoxyuridine, 3\u2032\u2010O\u2010acetyl\u2010N4\u2010benzoyl\u20102\u2032\u2010deoxycytidine, 3\u2032\u2010O\u2010levulinyl\u2010N4\u2010benzoyl\u20102\u2032\u2010deocytidine, and 5\u2032\u2010O\u2010levulinyl\u2010N4\u2010benzoyl\u20102\u2032\u2010deocytidine were purchased from Jena Bioscience, Jena, Germany.Restriction endonucleases, HindIII, BamHI, PstI, Phusion DNA polymerase, aLICator\u2122 LIC Cloning and Expression System Kit 3, PageRulerO\u2010acetyluridine and 2\u2032,3\u2032,5\u2032\u2010hexanoyluridine were prepared by adapting and modifying methods reported in the literature and evaporated under reduced pressure. The residue was purified by column chromatography to afford 2\u2032,3\u2032,5\u2032\u2010O\u2010triacetyluridine (1.45\u00a0g (95%) as a white solid. UV (CH3OH) \u03bbmax 257\u00a0nm. MS (ESI+): m/z 371.10 [M+H]+, 369.10 [M\u2212H]\u2212. 1H\u2010NMR (DMSO\u2010d6): \u03b4\u00a0=\u00a02.11 , 2.13 , 2.15 , 4.36 , 5.35 , 5.81 , 6.05 , 7.41 , 9.69 . 13C\u2010NMR (DMSO\u2010d6): \u03b4\u00a0=\u00a020.41, 20.51, 20.77, 63.18, 70.20, 72.72, 79.92, 87.46, 103.44, 139.36, 150.35, 163.00, 168.75, 169.67, 170.18. The NMR spectra were consistent with reported previously and sodium acetate was heated for 20\u00a0min at 90\u00b0C. Then uridine was added to the hot solution and stirred for 1\u00a0h. After the reaction was completed (TLC), the mixture was quenched with sodium bicarbonate and extracted with chloroform. The organic phase was dried and N,N\u2010dimethylpyridin\u20104\u2010amine were added. The reaction mixture was stirred for 3\u00a0h at room temperature. After the reaction was completed (TLC), the formed precipitate of N,N\u2032\u2010dicyclohexylurea was filtered, and the filtrate was concentrated under reduced pressure. The residue was treated with acetonitrile and the mixture of mono\u2010, di\u2010 and tri\u2010O\u2010hexanoyluridines precipitated as a white solid. The precipitate was filtered, dried to afford 0.5\u00a0g of hexanoyluridines. HPLC\u2010MS analysis indicated that the mixture consisted of 7% of mono\u2010O\u2010hexanoyluridine, 21% of di\u2010O\u2010hexanoyluridine and 72% of tri\u2010O\u2010hexanoyluridine. UV (CH3OH) \u03bbmax 256\u00a0nm. MS (ESI+): m/z 341.00 [M\u2212H]\u2212, 439.05 [M\u2212H]\u2212, 537.10 [M\u2212H].To a stirred mixture of uridine and triethylamine in 1,4\u2010dioxane (10\u00a0ml) hexanoic acid , R)\u20101\u2010phenylethanol or (S)\u20101\u2010phenylethanol and DMAP were dissolved in acetic anhydride and heated for 30\u00a0min at 55\u00b0C. The reaction mixture was monitored by thin\u2010layer chromatography . After the reaction was completed, the formed acetic acid was evaporated under reduced pressure. The residue was purified by column chromatography . After removal of solvents in vacuum, the desired products were obtained as colourless oils.Enantiomerically pure (Rf\u00a0=\u00a00.68 (CHCl3).1H NMR (CDCl3): \u03b4\u00a0=\u00a01.57 , 2.10 ; 5.92 , 7.27\u20137.44 . 13C NMR (CDCl3): \u03b4\u00a0=\u00a021.37, 22.22, 72.32, 126.11, 127.88, 128.51, 141.69, 170.34.Yield 261\u00a0mg (97%), colourless oil, Rf\u00a0=\u00a00.69 (CHCl3).1H NMR (CDCl3): \u03b4\u00a0=\u00a01.57 , 2.10 ; 5.91 , 7.27\u20137.44 . 13C NMR (CDCl3): \u03b4\u00a0 = \u00a021.37, 22.22, 72.32, 126.10, 127.88, 128.51, 141.69, 170.35.Yield 219\u00a0mg (81%), colourless oil, (R)\u20101\u2010phenylethyl acetate reported in literature \u20101\u2010phenylethanol or (S)\u20101\u2010phenylethanol and triethylamine were dissolved in dichloromethane (2\u00a0ml). After the solution was cooled to an ice bath, benzoyl chloride was added, and the reaction mixture was stirred for 1\u00a0h at ambient temperature. After the reaction was completed (TLC), the solvent was evaporated under reduced pressure. The residue was purified by column chromatography . The solvents were removed in vacuum, and the desired products were obtained as colourless oils.Enantiomerically pure (Rf\u00a0=\u00a00.75 (CHCl3).1H NMR (CDCl3): \u03b4\u00a0=\u00a01.71 , 6.17 , 7.31\u20137.52 , 8.10\u20138.16 . 13C NMR (CDCl3): \u03b4\u00a0=\u00a022.45, 72.94, 126.07, 127.92, 128.36, 128.58, 129.67, 130.54, 132.95, 141.81,165.84.Yield 184\u00a0mg (50%), colourless oil, Rf\u00a0=\u00a00.76 (CHCl3).1H NMR (CDCl3): \u03b4\u00a0=\u00a01.71 , 6.17 , 7.31\u20137.53 , 8.09\u20138.15 . 13C NMR (CDCl3): \u03b4\u00a0=\u00a022.44, 72.93, 126.07, 127.92, 128.36, 128.57, 129.67, 130.55, 132.95, 141.81,165.84.Yield 360\u00a0mg (97%), colourless oil, 1H NMR spectrum was consistent with spectrum of 1\u2010phenylethyl benzoate racemic ester reported previously \u20101\u2010phenylethanol or (S)\u20101\u2010phenylethanol , hexanoic acid , DCC and DMAP were dissolved in 10\u00a0ml of dichloromethane and stirred for 3\u00a0h at ambient temperature. After the reaction was completed (TLC), the formed precipitate was filtered, and the solvent was removed under reduced pressure. The residue was purified by column chromatography . After removal of solvents in vacuum, the desired products were obtained as colourless oils.Enantiomerically pure (Rf\u00a0=\u00a00.82 (CHCl3).1H NMR (CDCl3): \u03b4\u00a0=\u00a00.91 , 1.25\u20131.42 , 1.56 , 1.61\u20131.70 , 2.33\u20132.38 , 5.93 , 7.28\u20137.41 . 13C\u2010NMR (CDCl3): \u03b4\u00a0=\u00a013.91, 22.28, 22.32, 24.67, 31.27, 34.60, 72.01, 126.07, 127.79, 128.47, 141.88, 173.11.Yield 288\u00a0mg (80%), colourless oil, Rf\u00a0=\u00a00.82 (CHCl3).1H NMR (CDCl3): \u03b4\u00a0=\u00a00.91 , 1.27\u20131.40 , 1.56 , 1.61\u20131.70 , 2.32\u20132.38 , 5.93 , 7.28\u20137.42 . 13C\u2010NMR (CDCl3): \u03b4\u00a0 = \u00a013.91, 22.28, 22.32, 24.67, 31.27, 34.60, 72.01, 126.07, 127.79, 128.47, 141.88, 173.11.Yield 272\u00a0mg (75%), colourless oil, et\u00a0al., NMR spectra were consistent with spectra of 1\u2010phenylethyl hexanoate racemic ester reported in literature supplemented with the appropriate antibiotics , 1\u00a0mM IPTG and 1\u00a0mM X\u2010gal. Ten to twenty of individual white colonies\u2010forming clones were randomly chosen for plasmid DNA isolation by Thermo Scientific\u2122 GeneJET Plasmid Miniprep Kit (Thermo Fisher Scientific) and analysis of the length of the insert. The remaining undiluted mixture of the cells was spread on LB agar ) supplemented with 40\u00a0\u03bcg\u00a0ml\u22121 kanamycin and 100\u00a0\u03bcg\u00a0ml\u22121 ampicillin. After cultivation for 14\u201316\u00a0h, a biomass was scraped from agar, and total plasmid DNA was isolated by Thermo Scientific\u2122 GeneJET Plasmid Midiprep Kit (Thermo Fisher Scientific). The obtained DNA mixture (a metagenomics library) was stored at \u201320\u00b0C and used for a further transformation of the E.\u00a0coli DH10B \u0394pyrFEC::Km cells.Metagenomic libraries were constructed from soil and sediment samples using a pUC19 vector . For DNA\u22121 Na2HPO4 15\u00a0g\u00a0l\u22121 KH2PO4, 5\u00a0g\u00a0l\u22121 NH4Cl, 2.5\u00a0g\u00a0l\u22121 NaCl, 2% (w/v) glucose, 0.2% Casamino acid, 1\u00a0mM IPTG) containing 100\u00a0\u03bcg\u00a0ml\u22121 ampicillin and 40\u00a0\u03bcg\u00a0ml\u22121 kanamycin, 0.02\u00a0mg\u00a0ml\u22121 2\u2032,3\u2032,5\u2032\u2010O\u2010acetyluridine (1) or 2\u2032,3\u2032,5\u2032\u2010O\u2010hexanoyluridine (2) as the sole source of uridine, allowing only the growth of recombinants that can complement the uridine auxotrophy of the E.\u00a0coli DH10B \u0394pyrFEC::Km strain by hydrolysing the uridine esters.The clones exhibiting acetylesterase activity were identified on a MD medium . For further analysis the ORFs encoding putative hydrolases were chosen. When homology search did not predict a hydrolase, the deletion analysis to obtain the truncated variants of the plasmid and the functional reselection on the appropriate substrate was carried out. To confirm that the hits encoded the enzymes with an esterolytic activity, the selected genes were PCR\u2010amplified, and the resulting fragments were ligated into pET21a or pLATE31 expression vectors. Phylogenetic analysis was conducted using the Maximum Likelihood Tree routine of mega 7 software. and using the following sequencing primers: M13F\u2010pUC (5\u2032\u2010GTTTTCCCAGTCACGAC\u20103\u2032), M13R\u2010pUC (5\u2032\u2010CAGGAAACAGCTATGAC\u20103\u2032), T7 Promoter (5\u2032\u2010 TAATACGACTCACTATAGGG\u20103\u2032), T7 terminator (5\u2032\u2010TAATACGACTCACTATAGGG\u20103\u2032) or LIC Reverse Sequencing primer, 24\u2010mer (5\u2032\u2010GAGCGGATAACAATTTCACACAGG\u20103\u2032). Some individual clones contained more than one ORF in each DNA fragment. ORFs were analysed by using the Unipro UGENE program, and homology search was conducted using the Blast server (E.\u00a0coli cells transformed with recombinant plasmids were cultivated on LB agar supplemented with 100\u00a0\u03bcg\u00a0ml ampicillin.The genes of the selected enzymes were amplified with Phusion DNA polymerase using primers . MetagenE.\u00a0coli strain BL21 (DE3). E.\u00a0coli cells were grown in 20\u00a0ml BHI (Brain\u2010Heart\u2010Infusion Broth) medium containing ampicillin (100\u00a0mg\u00a0ml) at 37\u00b0C with aeration. Protein expression was induced by adding 0.5\u00a0\u03bcM IPTG at 0.6\u20131 OD600, and cells were grown for a further 4\u201318\u00a0h at 30\u00b0C. Cells expressing 12T and 3T were grown at 23\u00b0C after induction to increase protein solubility. Wet cell biomass from 20\u00a0ml culture broth were suspended in 5\u201310\u00a0ml of buffer A and disrupted by sonication for 2.5\u00a0min. A lysate was cleared by centrifugation at 15\u00a0000\u00a0\u00d7\u00a0g for 4\u00a0min. Cleared lysate was applied to 0.2\u00a0ml HisPur\u2122 Ni\u2010NTA spin column (equilibrated with the buffer A). The column was washed with the buffer A, and the proteins were eluted with buffer A containing 300\u00a0mM imidazole. The active fractions were combined and dialyzed against the buffer B , at 25\u00b0C. All the purification procedures were performed at room temperature.The recombinant proteins were overexpressed in The concentration of protein was determined using Pierce\u2122 Coomassie Plus (Bradford) Assay Reagent by Standard Microplate Protocol. Proteins were analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis according to Laemmli. Gels were developed in Coomassie Brilliant Blue G\u2010250 dye, scanned in 16 bit format and quantified by GelAnalyser program. Each sample contained 2\u00a0\u03bcg of total protein. Quantities of impurities and target proteins were estimated using calibration curve generated from known amounts of BSA: 0.125, 0.25 and 0.5\u00a0\u03bcg per band. The purity of analysed protein was calculated as the ratio between quantity of target protein and quantity of all proteins.et\u00a0al., et\u00a0al., \u03b5\u00a0=\u00a012.3\u00a0M\u22121\u00a0cm\u22121) per minute. Enzyme activity was tested against different pNP\u2010acyl esters .The activity of esterase were assayed by incubating the enzyme with 1\u00a0mM pNP\u2010substrate (from 10\u00a0mM stock in DMSO) in 50\u00a0mM potassium phosphate, pH 7.5, buffer at a 37\u00b0C for 10\u00a0min in 100\u00a0\u03bcl reaction volume. 5\u2013300\u00a0ng of protein, depending on the enzyme specificity for the substrate, were present into the reaction mixture. The absorbtion of the reaction mixture at 405\u00a0nm was measured against enzyme\u2010free blank to compensate for the substrate auto\u2010hydrolysis , pentaacetylglucose or pentaacetylgalactose (from 100\u00a0mM stock in acetone), 3\u2032\u2010O\u2010acetyl\u20102\u2032\u2010deoxyuridine, 3\u2032\u2010O\u2010acetyl\u2010N4\u2010benzoyl\u20102\u2032\u2010deoxycytidine, 3\u2032\u2010O\u2010levulinyl\u2010N4\u2010benzoyl\u20102\u2032\u2010deocytidine, and 5\u2032\u2010O\u2010levulinyl\u2010N4\u2010benzoyl\u20102\u2032\u2010deocytidine (100\u00a0mM stock in DMSO). The total reaction mix volume was 100\u00a0\u03bcl. Reaction mixture was incubated at room temperature up to 5\u00a0h. A change of colour from red to yellow indicated the hydrolysis of esters , 1\u00a0\u03bcl enzyme (0.1\u20134.6\u00a0\u03bcg/reaction) and 10\u00a0mM substrate: d\u2010glucose pentaacetate, \u03b2\u2010d\u2010 galactose pentaacetate (from 100\u00a0mM stock in acetone), 3\u2032\u2010O\u2010benzoyl\u20102\u2032deoxyuridine (10\u00a0mg\u00a0ml\u22121 stock in DMF), 3\u2032\u2010O\u2010levulinyl\u2010N4\u2010benzoyl\u20102\u2032deocytidine and 5\u2032\u2010O\u2010levulinyl\u2010N4\u2010benzoyl\u20102\u2032\u2010deocytidine (from 100\u00a0mM stock in DMSO). The total reaction mix volume was 20\u00a0\u03bcl. Reaction mixture was incubated at 30\u00b0C temperature up to 3\u00a0h. Thin\u2010layer chromatography (TLC) was conducted on the Merck silica gel 60F254 plates, using the dichloromethane and methanol (9:1) mixture of solvents. \u03b2\u2010d\u2010glucose pentaacetate and \u03b2\u2010d\u2010 galactose pentaacetate were visualized by anisaldehide stain . The plate was developed by heating on a hot plate. Synthetic nucleosides were exposing to UV light.A hydrolytic activity was assayed in reaction mixture containing 45\u00a0mM potassium phosphate buffer, pH 7.5, 1\u00a0\u03bcl enzyme (0.1\u20134.6\u00a0\u03bcg/reaction) and 10\u00a0mM substrate: \u03b2\u2010There are no conflicts to declare.Table\u00a0S1. Metagenomic libraries used in this work.Table\u00a0S2. Primers used for amplification of genes of selected hydrolases.Table\u00a0S3. Predicted signal peptide sequences identified in the selected esterases by bioinformatics.Fig.\u00a0S1. Selected clones on MD medium, MD+ compound 1, MD+compound 2 and MD+uridine after 2\u00a0days of incubation at 37\u00b0C.Fig.\u00a0S2. The alignment was performed by ClustalW software.Fig.\u00a0S3. Analysis of the purified esterases by SDS\u2010PAGE.Fig.\u00a0S4. LB medium with tributyrin plates after 2\u00a0days of incubation at 37\u00b0C.Scheme\u00a0S1. Synthesis of optically active esters (3\u20138).Click here for additional data file."} +{"text": "I ion is coordinated by two heterocyclic N atoms from two ligands in a linear configuration, forming a discrete coordination complex. There is an O\u2014H\u22efO hydrogen bond between 2-tza\u2212 and 2tzaH of adjacent complexes. The hydrogen atom is shared between the two oxygen atoms.The Ag 4H2NO2S)(C4H3NO2S)] or [Ag(2-Htza)(2-tza)] is reported . The AgI ion is coordinated by two heterocyclic N atoms from two ligands in a linear configuration, forming a discrete coordination complex. There is an O\u2014H\u22efO hydrogen bond between 2-tza\u2212 and 2tzaH of adjacent complexes. The hydrogen atom is shared between the two oxygen atoms. This inter\u00adaction produces a hydrogen-bonded tape parallel to the [110] direction, which is augmented through inter\u00admolecular C\u2014H\u22efO hydrogen-bonding inter\u00adactions between the bound thia\u00adzole groups. There is a further rather long Ag\u22efO inter\u00adaction that assembles these tapes into columns, between which there are C\u2014H\u22ef\u03c0 inter\u00adactions, leading to the formation of a three-dimensional supra\u00admolecular architecture.The linear two-coordinate silver (I) complex [Ag(C The whole mol\u00adecular structure can be generated by an inversion centre; the AgI atom is located at the 2a Wyckoff position (I centre shows a linear coordin\u00adation with two 2-tza ligands coordinating through the heterocyclic N atoms with an Ag\u2014N bond length of 2.1463\u2005(14)\u2005\u00c5. Statistically one of these ligands has an appended carb\u00adoxy\u00adlic acid and the other a carboxyl\u00adate. A rather long Ag\u22efO2 inter\u00adaction is also observed with the distance of 2.8401\u2005(13)\u2005\u00c5. This is significantly larger than the mean value [2.54\u2005(11)\u2005\u00c5] of the Ag\u22efO=C distances in the Cambridge Database (2-tza\u2212). The carboxyl\u00adate is located close to a second symmetry-equivalent carboxyl\u00adate generated by the symmetry operation 1\u00a0\u2212\u00a0x, \u2212y, 1\u00a0\u2212\u00a0z. Statistically, one of these two groups is protonated. The close approach facilitates the formation of a linear hydrogen bond between them of an adjacent discrete mol\u00adecule (Table\u00a01a). This tape is inclined at an angle of 55.278\u2005(19)\u00b0 with the (001) plane. Each tape is connected through an Ag\u22efO1 inter\u00adaction [d(Ag\u22efO1) = 2.9606\u2005(14)\u2005\u00c5], generating columns of complex mol\u00adecules along the [100] direction . These columns are further linked via C\u2014H\u22ef\u03c0 inter\u00adactions between the thia\u00adzole rings and 2-Htza were dissolved in 5.0\u2005mL of deionized water in a small vial (ca 16\u2005mm in diameter). The vial was left undisturbed at ambient temperature for three days during which colourless block-shaped crystals of the title compound crystallized and were isolated for X-ray data collection.AgNOUiso(H) = 1.5Ueq(O). The C-bound H atoms were refined isotropically, with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019000124/nk2248sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019000124/nk2248Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019000124/nk2248Isup3.molSupporting information file. DOI: 1888609CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Polynitrile anions are known for their ability to combine with transition metals and co-ligands to form ternary systems. Here we report on the crystal structure of tris\u00ad(2\u20132\u2032-bi\u00adpyridine)\u00adcobalt(II) bis\u00ad. 10H8N2)3](C9H5N4O)2, the tris\u00ad\u00adcobalt(II) dication lies across a twofold rotation axes in the space group C2/c. The N atoms of the three bi\u00adpyridine ligands form a distorted octa\u00adhedron around the cobalt ion. All the N atoms of the polynitrile 1,1,3,3-tetra\u00adcyano-2-eth\u00adoxy\u00adpropenide anions participate in C\u2014H\u22efN hydrogen bonds ensuring crystal cohesion and forming a three-dimensional structure. The structure is further stabilized by C\u2014H\u22ef\u03c0(cation) and anion\u22ef\u03c0(cation) inter\u00adactions.In the title compound, [Co(C The cobalt ion is ligated by the N atoms of the 2,2\u2032-bi\u00adpyridine ligands forming a slightly distorted octa\u00adhedral coordination sphere; the Co1\u2014N bond lengths vary from 2.122\u2005(3) to 2.148\u2005(3)\u2005\u00c5. In the bpy unit, the pyridine rings are inclined to each other by 10.40\u2005(16)\u00b0, while in the other bpy unit (involving atom N3) bis\u00adected by a twofold rotation axis the pyridine rings are coplanar. The observed distortion of the CoIIcoordination sphere is probably the consequence of the hydrogen bonding between the [Co(C10H8N2)3]2+ cation and the flexible tcnoet\u2212 anion (see Supra\u00admolecular features).The asymmetric unit of the title compound (I)\u2212 anion, the six central C\u2014C distances within the anion range from 1.382\u2005(4) to 1.429\u2005(5)\u2005\u00c5 while the C\u2261N distances vary from 1.138\u2005(4) to 1.151\u2005(4)\u2005\u00c5 \u00b0.In the tcnoet\u00c5 Table\u00a01. As obseThe crystal packing of (I)\u2212 anions linked by eight C\u2014H\u22efN hydrogen bonds as shown in Fig.\u00a03\u2212 anions are doubly connected to the cationic units via C8\u2014H8\u22efN7iii; C11i\u2014H11i\u22efN6iii and their symmetric C8i\u2014H8i\u22efN7iv; C11\u2014H11\u22efN6iv. Four tcnoet\u2212 anions are linked to atoms N4 and N5 via C7\u2014H7\u22efN4iv, C2\u2014H2\u22efN5v, C7i\u2014H7i\u22efN4ii and C2i\u2014H2i\u22efN5vii. One of the anions plays the role of a donor in the structural linkage. Indeed, one tcnoet\u2212 anion is linked by an N\u22efH\u2014C inter\u00adaction to the same [Co(C10H8N2)3]2+ unit (via N6\u22efH11iii\u2014C11iii and N7\u22efC8ii\u2014H8ii) and to two other cationic units by N4\u22efH7i\u2014C7i and N5\u22efH2iv\u2014C2iv inter\u00adactions. This environment where the negative charge is delocalized over the central propenide unit as well as into the cyano groups is illustrated in Fig.\u00a04The cations are surrounded by six tcnoetB\u22efCg1vi; see Table\u00a02Cg2; see Table\u00a02The crystal structure of (I)et al., 2016\u2212 anion associated with an organic cation to form a salt-like compound the water mol\u00adecule links the tcnoet\u2212 anion and the iron aggregate via O\u2014H\u22efN hydrogen bonds, forming chains, whereas in (I)ca 28.1\u00b0 compared to 31.7\u2005(3)\u00b0 in (I)A search in the Cambridge Structural Database , 2,2-bi\u00adpyridine and K(tcnoet) in water\u2013ethanol . This mixture was sealed in a Teflon-lined autoclave and held at 423\u2005K for three days, and then cooled to ambient temperature at a rate of 10\u2005K\u2005h\u22121 (yield: 54%). Colourless plate-like crystals of the title compound were selected directly from the synthesized product.The title compound was synthesized solvothermally under autogenous pressure from a mixture of CoNOUiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018018261/su5465sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989018018261/su5465Isup2.hklStructure factors: contains datablock(s) I. DOI: 1887084CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "GBA) is a lysosomal \u03b2\u2010glucosidase\u2010degrading glucosylceramide. Its deficiency causes Gaucher disease (GD), a common lysosomal storage disorder. Carrying a genetic abnormality in GBA constitutes at present the largest genetic risk factor for Parkinson's disease (PD). Conduritol B epoxide (CBE), a mechanism\u2010based irreversible inhibitor of GBA, is used to generate cell and animal models for investigations on GD and PD. However, CBE may have additional glycosidase targets besides GBA. Here, we present the first in\u00a0vivo target engagement study for CBE, employing a suite of activity\u2010based probes to visualize catalytic pocket occupancy of candidate off\u2010target glycosidases. Only at significantly higher CBE concentrations, nonlysosomal glucosylceramidase (GBA2) and lysosomal \u03b1\u2010glucosidase were identified as major off\u2010targets in cells and zebrafish larvae. A tight, but acceptable window for selective inhibition of GBA in the brain of mice was observed. On the other hand, cyclophellitol, a closer glucose mimic, was found to inactivate with equal affinity GBA and GBA2 and therefore is not suitable to generate genuine GD\u2010like models.Glucocerebrosidase (http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/2/1/45.html), nonlysosomal \u03b2\u2010glucocerebrosidase (http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/2/1/45.html); cytosolic \u03b2\u2010glucosidase (http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/2/1/21.html); \u03b1\u2010glucosidases (http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/2/1/20.html); \u03b2\u2010glucuronidase (http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/2/1/31.html).Glucocerebrosidase ( In\u00a0vivo target engagement of mechanism\u2010based glucocerebrosidase (GBA) inhibitors\u2014conduritol B epoxide (CBE) and cyclophellitol (CP)\u2014were examined in cultured cells, zebrafish larvae and mice by competitive activity\u2010based protein profiling (ABPP). This method utilizes suicide fluorescent enzyme reporter molecules to assess active site occupancy of target glycosidases by inhibitors. The in\u00a0vivo targets of CBE and CP and their selectivity towards GBA were revealed. ABPactivity\u2010based probeABPPactivity\u2010based protein profilingCBEconduritol B epoxideCPcyclophellitoldpfdays postfertilizationGBAglucocerebrosidaseGDGaucher diseaseGlcSphglucosylsphingosinePDParkinson's diseasehttp://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/2/1/45.html) is a retaining \u03b2\u2010glucosidase that degrades the glycosphingolipid, glucosylceramide. Inherited deficiencyof GBA is the cause of autosomal recessive Gaucher disease (GD) The lysosomal enzyme glucocerebrosidase Fig.\u00a0A 7, 8, 9m to 100\u00a0mm CBE for 2\u00a0h up to 60\u00a0days \u22121 body weight during 2\u00a0h up to 36\u00a0days in\u00a0vitro inhibition of retaining \u03b1\u2010glucosidases (http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/2/1/20.html) in\u00a0vitroin\u00a0situhttp://www.chem.qmul.ac.uk/iubmb/enzyme/EC3/2/1/45.html), and inhibition of the lysosomal \u03b2\u2010glucuronidase in mice in\u00a0vivo towards GBA2 and other glycosidases is unknown.A major advantage of CBE's pharmacological use in cultured cells and mice is its tunability: the extent of GBA inactivation can be adjusted by variation in the inhibitor concentration and/or exposure time in\u00a0vivo selectivity of CBE and CP in cells and animal models. We envisioned that besides the traditional enzymatic assays employing fluorogenic substrates, activity\u2010based probes (ABPs) could be superior tools for this study. Unlike enzymatic substrate assays, which do not easily distinguish similar enzymatic activities such as GBA vs GBA2, ABPs would allow direct and unambiguous visualization of respective target glycosidases that are not occupied/inactivated by CBE or CP at the active site pocket. Cravatt and coworkers and van der Stelt and colleagues earlier used ABPs directed towards proteases and lipases in a competitive activity\u2010based protein profiling (ABPP) approach to identify in\u00a0vivo target engagement of small compounds 3\u20136). Here, we report a detailed in\u00a0vivo target engagement study for CBE and CP. Through parallel application of both the competitive ABPP method and enzymatic assay in lysates of cultured cells, zebrafish (Danio rerio) larvae, and brains of mice treated in\u00a0vivo with a relevant range of concentrations of CBE or CP, we have systematically assessed their in\u00a0vivo off\u2010targets and selectivity windows for GBA.Our aim was to systematically study the 3 and 4) 1 and CP 2 with a competitive ABPP method by which the irreversible occupancy of the catalytic nucleophile of GBA by the inhibitors during preincubation is assessed. As a validation, competition of ABP labelling by CBE and CP was compared to the loss of GBA activity measured using 3.75\u00a0mm 4\u2010methylumbelliferyl\u2010\u03b2\u2010glucoside as substrate 4c was quantified by SDS/PAGE and fluorescence scanning. IC50 values (concentrations of inhibitor yielding a 50% reduction of ABP 4c labelling) were determined and found to be 26.6\u00a0\u03bcm at 30\u00a0min CBE preincubation, and 2.30\u00a0\u03bcm at 180\u00a0min preincubation and GBA2 (110\u00a0kDa) following SDS/PAGE analysis. First, we studied HEK293T cells expressing GBA2. Cells were exposed for 24\u00a0h to different concentrations of CBE (0.1\u00a0\u03bcm\u201310\u00a0mm) after which the residual amount of GBA, GBA2, GAA, GANAB and GUSB in cell lysates that can still be labelled with the appropriate ABPs , GBA2 (IC50\u00a0=\u00a0315\u00a0\u03bcm), GAA (IC50\u00a0=\u00a0249\u00a0\u03bcm), GANAB (IC50\u00a0=\u00a02900\u00a0\u03bcm) and GUSB (IC50\u00a0= 857\u00a0\u03bcm). , GBA2 (IC50 =272\u00a0\u03bcm), GAA (IC50 = 309\u00a0\u03bcm), GANAB (IC50 = 1580\u00a0\u03bcm) and GUSB (IC50 = 607\u00a0\u03bcm) for their possible inactivation by CBE. At the highest concentration of CBE (10\u00a0mm) tested, no significant loss of activity was observed for any of these additional lysosomal enzymes and lysosomal \u03b2\u2010glucuronidase GUSB. For each of these enzymes, ABPs have been designed, and enzymatic activity assays with fluorogenic substrates established and GBA2 10\u00a0kDa fotive GBA 8\u201366\u00a0kDa m\u2013100\u00a0mm) in the egg water for 5\u00a0days. The larvae were collected, lysed and analysed by the competitive ABPP method. Exposure to 100\u00a0mm CBE was found to reduce ABP labelling of all five glycosidases , GBA2 (IC50 = 890\u00a0\u03bcm), GAA (IC50 = 9550\u00a0\u03bcm), GANAB (IC50 = 4700\u00a0\u03bcm) and GUSB (IC50 = 6470\u00a0\u03bcm) except for \u03b2\u2010galactosidase (IC50 = 11.2\u00a0mm) and \u03b1\u2010galactosidase (IC50 = 22.5\u00a0mm) to CBE Table\u00a0. Thus, im) Table\u00a0, Fig.\u00a07.in\u00a0vivo targets of CBE in brain of mice treated daily from day 8 to either day 25 with 37.5\u00a0mg CBE\u00b7kg\u22121 body weight or to day 14 with 100\u00a0mg CBE\u00b7kg\u22121 body weight \u22121 body weight for 24\u00a0h. IC50 values of CP for blocking ABP labelling were 0.063\u00a0\u03bcm for GBA and 0.154\u00a0\u03bcm for GBA2 Fig.\u00a0A. As detm) also comparably competed GBA and GBA2 labelling, but not that of GUSB, GAA and GANAB 4m ABP 4c. We therefore generated HEK293T cells overexpressing GBA3 to study the inhibitor sensitivity of the enzyme. In these cells we could measure the in\u00a0vivo interaction of CBE and CP with GBA3 by ABP detection (ABP 4c) by acid ceramidase\u2010mediated conversion of accumulating GlcCer in lysosomes m CBE; sixfold increase at 10\u00a0\u03bcm CP) and in zebrafish larvae . Thus, a selective window for in\u00a0vivo GBA inactivation by CBE exists Fig.\u00a0. ImportaOur investigation also revealed that CP is a more potent GBA inactivator. However, CP inhibits GBA2 on a par with GBA in both cultured cells and zebrafish larvae Table\u00a0, and the2 (fibroblasts) or 7% CO2 (HEK293T). Zebrafish (Danio rerio) were handled and maintained according to standard protocols (zfin.org). Adult zebrafish were housed at a density of 40 per tank, with a cycle of 14\u00a0h of light and 10\u00a0h of darkness. Adults, embryos and larvae were kept at a constant temperature of 28.5\u00a0\u00b0C. Embryos and larvae were raised in egg water . Synchronized wild\u2010type ABTL zebrafish embryos were acquired after mating of single male and female couples (both > 3\u00a0months old). Frozen brain samples from wild\u2010type and CBE\u2010treated mice were obtained from a previous study Cyclophellitol (CP) and the ABPs were synthesized as described earlier CO2 HEK23T. Zebram citric acid\u2014Na2HPO4, 0.1% (w/v) bovine serum albumin) for a period of 30\u00a0min to 2\u00a0h. After stopping the substrate reaction with 200\u00a0\u03bcL 1M NaOH\u2010glycine (pH 10.3), 4MU\u2010emitted fluorescence was measured with a fluorimeter LS55 using \u03bbEX 366\u00a0nm and \u03bbEM 445\u00a0nm prism 7.0 by the [inhibitor] vs response\u2014various slope (four parameters) method to obtain IC50 values. The substrate mixtures used for each enzyme are listed as follows: GBA, 3.75\u00a0mm 4MU\u2010\u03b2\u2010D\u2010glucopyranoside at pH 5.2, supplemented with 0.2% (w/v) sodium taurocholate and 0.1% (v/v) Triton X\u2010100, and 25\u00a0nm N\u2010(5\u2010adamantane\u20101\u2010yl\u2010methoxy\u2010pentyl)\u2010deoxynojirimycin (AMP\u2010DNM), a GBA2\u2010specific inhibitor m 4MU\u2010\u03b2\u2010D\u2010glucopyranoside at pH 5.8, with pre\u2010incubation with 1\u00a0\u03bcm ABP 3a for 30\u00a0min to specifically inhibit GBA activity; \u03b1\u2010glucosidases, 3\u00a0mm 4MU\u2010\u03b1\u2010D\u2010glucopyranoside at pH 4.0 (GAA) or at 7.0 (GANAB), GUSB, 2\u00a0mm 4MU\u2010\u03b2\u2010D\u2010glucuronide at pH 5.0; \u03b1\u2010galactosidases, 2\u00a0mm 4MU\u2010\u03b1\u2010D\u2010galactopyranoside at pH 4.6; \u03b2\u2010galactosidases, 1\u00a0mm 4MU\u2010\u03b2\u2010D\u2010galactopyranoside at pH 4.3 with 0.2\u00a0M NaCl; \u03b1\u2010mannosidases, 10\u00a0mm 4MU\u2010\u03b1\u2010D\u2010mannopyranoside at pH 4.0; \u03b2\u2010mannosidases, 2\u00a0mm 4MU\u2010\u03b2\u2010D\u2010mannopyranoside (Glycosynth) at pH 4.2; \u03b2\u2010hexosaminidase HexA, 5\u00a0mm 4MU\u2010\u03b2\u2010D\u20106\u2010sulpho\u20102\u2010acetamido\u20102\u2010deoxy\u2010glucopyranoside at pH 4.4; \u03b2\u2010hexosaminidases HexA/B, 5\u00a0mm 4MU\u2010\u03b2\u2010N\u2010acetyl\u2010glucosaminide at pH 4.5; \u03b1\u2010N\u2010acetyl\u2010galactosaminidase, 1\u00a0mm 4MU\u2010\u03b1\u2010N\u2010acetyl\u2010galactosaminide at pH 4.5; \u03b1\u2010L\u2010fucosidase, 1\u00a0mm 4MU\u2010\u03b1\u2010L\u2010fucopyranoside at pH 5.0, \u03b1\u2010L\u2010iduronidase, 2\u00a0mm 4MU\u2010\u03b1\u2010L\u2010iduronide (Glycosynth) at pH 4.0; GBA3, 3.75\u00a0mm 4MU\u2010\u03b2\u2010D\u2010glucopyranoside at pH 6.0.All assays were performed in 96\u2010well plates at 37\u00a0\u00b0C for human, zebrafish and mice material. Samples were diluted with McIlvaine buffer to a final volume of 25\u00a0\u03bcL, at pH appropriate for each enzyme. Assays were performed by incubating the samples with 100\u00a0\u03bcL 4MU\u2010 (4\u2010methylumbelliferyl\u2010) substrates diluted in McIlvaine buffer Triton\u2010100, 0.2% (w/v) sodium taurocholate), or labelled together with GBA2 using 200\u00a0nm \u03b2\u2010aziridine ABP 4c at pH 5.5. GBA2 was labelled with 200\u00a0nm \u03b2\u2010aziridine ABP 4a, 4b or 4c. The \u03b1\u2010glucosidases GAA and GANAB were first preincubated with 200\u00a0nm ABP 4a for 30\u00a0min (pH 4.0 for GAA and pH 7.0 for GANAB), followed by labelling with 500\u00a0nm ABP 6a or 6c at pH 4.0 or 7.0. The \u03b2\u2010glucuronidase GUSB was preincubated with 200\u00a0nm ABP 4a for 30\u00a0min, followed by labelling with 200\u00a0nm \u03b2\u2010aziridine ABP 5c. After ABP incubation, proteins were denatured by boiling the samples with 5\u00d7 Laemmli buffer (50% (v/v) 1\u00a0M Tris\u2010HCl, pH 6.8, 50% (v/v) 100% glycerol, 10% (w/v) DTT, 10% (w/v) SDS, 0.01% (w/v) bromophenol blue) for 5\u00a0min at 98\u00a0\u00b0C, and separated by electrophoresis on 7.5% or 10% (w/v) SDS/PAGE gels running continuously at 90\u00a0V and curve\u2010fitted using prism 7.0 (GraphPad Software). After fluorescence scanning, SDS/PAGE gels were stained for total protein with Coomassie G250 and scanned on a ChemiDoc MP imager .Residual active, not irreversibly inhibited glycosidases were labelled with excess fluorescent ABPs in the optimum McIlvaine buffer, if not otherwise stated (see above). ABP labelling was performed at 37\u00a0\u00b0C for 30\u00a0min for all materials, in\u00a0a total sample volume of 20\u201340\u00a0\u03bcL and 0.5\u20131% DMSO concentration. GBA was labelled with 200\u00a0ny at 90\u00a0V, 37, 38.50 measurements using enzymatic assay, 3.16\u00a0ng (53 fmole) of rGBA was prepared in 12.5\u00a0\u03bcL McIlvaine buffer supplemented with 0.1% (v/v) Triton X\u2010100, and 0.2% (w/v) sodium taurocholate, and incubated with 12.5\u00a0\u03bcL of inhibitors (CBE or CP) diluted in McIlvaine buffer at 37\u00a0\u00b0C for various time periods. Residual activity of rGBA was measured as described in previous section. For assessing the occupancy of active site pocket by the inhibitors using ABP labelling, the same amount of rGBA was prepared in 10\u00a0\u03bcL of the same McIlvaine buffer, incubated firstly with 2.5\u00a0\u03bcL inhibitor dilutions prepared in McIlvaine buffer at 37\u00a0\u00b0C for various time periods, then with 2.5\u00a0\u03bcL ABP dilutions prepared in McIlvaine buffer; detection of ABP\u2010labelled rGBA follows the procedures described in the previous section.For ICm) or CP (0.001\u201310\u00a0\u03bcm) for 24\u00a0h at 37\u00a0\u00b0C. In addition, confluent human fibroblasts were similarly treated in 15\u2010cm dishes for 2, 24 and 72\u00a0h. For lysis, cells were washed three times with PBS, subsequently lysed by scraping in potassium phosphate buffer , aliquoted and stored at \u221280\u00a0\u00b0C. After determination of the protein concentration, lysates containing equal protein amount were adjusted to 12\u00a0\u03bcL with potassium phosphate buffer and subjected to residual activity measurements using enzymatic assay or ABP detection . For in\u00a0vivo ABP labelling, HEK293T cells expressing GBA2 were incubated with culture medium containing CBE (0.015\u201315\u00a0000\u00a0\u03bcm) or CP (0.001\u201310\u00a0\u03bcm) for 24\u00a0h. Medium was removed and cells were incubated for 4\u00a0h with culture medium containing a mixture of 200\u00a0nm ABP 4b, 1\u00a0\u03bcm ABP 5c and 500\u00a0nm ABP 6c, or with DMSO only (negative control). Lysates were prepared and measured for protein concentration as described above, and samples containing 60\u00a0\u03bcg total protein were subjected to ABP detection .Confluent HEK293T stably expressing human GBA2 were cultured in 12\u2010well plates in triplicates with(out) CBE , or CP (0.001\u2013100\u00a0\u03bcm) for 120\u00a0h at 28.5\u00a0\u00b0C. Per condition, n = 48 embryos were used. At 120\u00a0h (5 dpf), larvae were collected, rinsed three times with egg water, fully aspirated, snap\u2010frozen in liquid nitrogen and stored at \u221280\u00a0\u00b0C until homogenization in 200\u00a0\u03bcL 25\u00a0mm potassium phosphate buffer per 48 individual. Lysis was conducted by sonication with a Polytron PT 1300D sonicator on ice at 20% power for 3\u00a0s, and repeated three times. Samples containing 20\u201345\u00a0\u03bcg total protein were subjected to ABP detection or enzymatic assay.Adult zebrafish were not sacrificed for this study; all experiments were performed on embryos/larvae before the free\u2010feeding stage and did not fall under animal experimentation law according to the EU Animal Protection Directive 2010/63/EU. For \u22121 body weight, or PBS, from day 8 until day of sacrifice as previously described m potassium phosphate buffer (4\u00d7 volume/wet tissue weight) with 1.0\u00a0mm glass beads using a Fastprep\u201024 instrument set at 6\u00a0m\u00b7s\u22121 for 20\u00a0s, repeated three times, while chilling samples on ice for 2\u00a0min between separate runs. Crude lysates were isolated from the glass beads by pipetting into sterile Safe\u2010Lock Eppendorf tubes. Homogenates were measured for protein concentration, aliquoted and snap\u2010frozen in liquid nitrogen. Samples containing 50\u00a0\u03bcg total protein were subjected to ABP detection or enzymatic assay. Two\u2010tailed unpaired t\u2010test was performed in prism 7.0 software (GraphPad Software) to derive statistical significance, where P\u00a0<\u00a00.05 was considered significant.Brain hemispheres were obtained from mice injected daily with CBE at either 37.5\u00a0mg or 100\u00a0mg\u00b7kgm ABP 4c at pH 6.0. For enzymatic assay, lysates were separated from GBA2 and GBA by centrifugation (16\u00a0000\u00a0g for 10\u00a0min at 4\u00a0\u00b0C) and incubation of the resulting supernatant with 20\u00a0\u03bcL concanavalin A sepharose beads (Sigma\u2010Aldrich) in 200\u00a0\u03bcL of binding buffer for 1\u00a0h at 4\u00a0\u00b0C on a rotor. Next,\u00a0beads were removed from the supernatant by centrifugation. Five microlitres of the supernatant was added with 20\u00a0\u03bcL McIlvaine buffer (pH 6.0), and subjected to enzymatic assays for GBA3 activity. Measured activity was\u00a0normalized with the corresponding protein concentration of each sample, and data were processed as earlier described.HEK293T cells expressing human GBA3 were cultured, treated (triplicate sets) and lysed in an identical setup as described with GBA2\u2010expressing HEK293T cells. Lysates (12\u00a0\u03bcg protein) were subjected to ABP detection using 200\u00a0nm) or CP (0.01\u201310\u00a0\u03bcm)) for 112\u00a0h at 28\u00a0\u00b0C. Thereafter, zebrafish larvae were washed three times with egg water, and collected in clean screw\u2010cap Eppendorf tubes (three tubes of three larvae per inhibitor concentration). Lipids were extracted and measured according to methods described previously 13C\u2010GlcSph \u22121 in methanol (MeOH), 480\u00a0\u03bcL MeOH and 250\u00a0\u03bcL CHCl3 were added to the sample, stirred, incubated for 30\u00a0min at RT, sonicated (5\u00a0\u00d7\u00a01\u00a0min in sonication water bath) and centrifuged for 10\u00a0min at 15\u00a0700\u00a0g. Supernatant was collected in a clean tube, 250\u00a0\u03bcL CHCl3 and 450\u00a0\u03bcL 100\u00a0mm formate buffer (pH 3.2) were added. The sample was stirred and centrifuged, the upper phase was transferred to a clean tube. The lower phase was extracted with 500\u00a0\u03bcL MeOH and 500\u00a0\u03bcL formate buffer. The upper phases were pooled and taken to dryness in a vacuum concentrator at 45\u00a0\u00b0C. The residue was extracted with 700\u00a0\u03bcL butanol and 700\u00a0\u03bcL water, stirred and centrifuged. The upper phase (butanol phase) was dried and the residue was dissolved in 100\u00a0\u03bcL MeOH. Ten microlitres of this sample was injected to the LC\u2010MS for lipid measurement. Two\u2010tailed unpaired t\u2010test was performed in Prism 7.0 software (GraphPad Software) to derive statistical significance, where P\u00a0<\u00a00.05 was considered significant.Zebrafish embryos at 8\u00a0h postfertilization were seeded in 12\u2010well plates and treated with CBE (10\u20131000\u00a0\u03bcThere are no conflicts of interest declared.CLK, WWK and IZ performed the experiments; LTL, AV, AHF, AHM and HPS provided essential materials. CLK, WWK, and MM analysed the data. WWK and MM helped to write the manuscript. CLK, JMFGA and MA wrote the manuscript. AHF, HSO and MA revised the manuscript. HSO and JMFGA conceived and designed the study."} +{"text": "III ion is nine-coordinated in a distorted tricapped trigonal\u2013prismatic geometry by three oxygen atoms and three nitro\u00adgen atoms from three benzhydrazide (bzz) ligands, one oxygen atom from the isophthalate (itp2\u2212) ligand, and two oxygen atoms from coordinated water mol\u00adecules. The crystal structure features extensive hydrogen bonding as well as C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions.The first benzohydrazide complex of a lanthanide is reported. The Sm 8H4O4)(C7H8N2O)3(H2O)2]NO3, systematic name di\u00adaqua\u00adtris\u00adsamarium(III) nitrate, the SmIII ion is nine-coordinated in a distorted tricapped trigonal\u2013prismatic geometry by three oxygen atoms and three nitro\u00adgen atoms from three benzhydrazide (bzz) ligands, one oxygen atom from the isophthalate (itp2\u2212) ligand, and two oxygen atoms from coordinated water mol\u00adecules. The nitrate group is disordered over two sets of sites with occupancy factors of 0.310\u2005(17) and 0.690\u2005(17). In the crystal, adjacent mol\u00adecules are linked into chains via pairs of O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds between the carboxyl\u00adate acceptor and the coordinated water and amine NH2 donors. Mol\u00adecules are further stacked by \u03c0\u2013\u03c0 inter\u00adactions involving the benzene ring of the itp2\u2212 ligands, forming double chains that extend in the b-axis direction. These double chains are further linked into a three-dimensional supra\u00admolecular network via hydrogen bonds between the complex mol\u00adecule and the nitrate groups along with C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions involving the benzene rings of the bzz and itp2\u2212 ligands.The first benzohydrazide complex of a lanthanide is reported. In the title compound, [Sm(C Distinct from transition metal centers, lanthanide ions often demonstrate high and variable coordination numbers as well as diverse coordination geometries, which can lead to versatile structures and topologies ligands, one completely deprotonated isophthalate (itp2\u2212) ligand, two coordinated water mol\u00adecules, and one disordered NO3\u2212 ion. The hydrazide group of the bzz ligand adopts a bidentate \u03bc2-\u03b71:\u03b71 chelating coordination mode, whereas the carboxyl\u00adate groups of the fully deprotonated itp2\u2212 ligand display a \u03bc1-\u03b71:\u03b70 monodentate coordination fashion. The SmIII ion is nine-coordinated by three oxygen atoms and three nitro\u00adgen atoms of three different bzz ligands, one oxygen atom (O4) from the completely deprotonated itp2\u2212 ligand, and other two oxygen atoms from the coordinated water mol\u00adecules. The central metal SmIII atom can be described as having a distorted tricapped trigonal\u2013prismatic geometry, Fig.\u00a02III complexes \u2005\u00c5 and a dihedral angle = 0.0\u2005(2)\u00b0 , and the bzz and itp2\u2212 ligands , are also observed, which help further to stabilize the crystal structure.As can be seen in Fig.\u00a03s Table\u00a01 involvinet al., 2016et al., 1971et al., 2003et al., 2011et al., 2009A search of the Cambridge Structural Database 3\u00b76H2O , bzz , H2itp , and H2O (4\u2005ml) was sealed in a 15\u2005ml Teflon-lined steel autoclave and heated at 373\u2005K for 24\u2005h. The mixture was cooled to room temperature and light-yellow block-shaped crystals of the title compound were obtained in 79% yield . Analysis calculated (%) for C29H32N7O12Sm (1376.80): C 42.43; H 3.93; N 11.94%. Found: C 42.46; H 3.96; N 11.90%.A mixture of Sm(NOUiso(H) = 1.2Ueq(C). The H atoms bonded to O and N atoms were located in a difference-Fourier map, but were refined with distance restraints of O\u2014H = 0.84 \u00b1 0.01\u2005\u00c5 and N\u2014H = 0.88 \u00b1 0.01\u2005\u00c5, and with Uiso(H) = 1.5Ueq(O) and 1.2Ueq(N). The nitrate group is disordered over two sets of sites, with occupancy factors of 0.310\u2005(17) and 0.690\u2005(17).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018015360/pj2059sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018015360/pj2059Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018015360/pj2059Isup3.cdxSupporting information file. DOI: 1876239CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S,3S)-2-amino-3-methyl\u00adpenta\u00adnoic acid, l-isoleucine C6H13NO2, crystallizes in the monoclinic space group P21 with four independent mol\u00adecules in the asymmetric unit. In the crystal, N\u2013H\u22efO hydrogen bonds link two pairs of independent mol\u00adecules and their symmetry-related counterparts to form two types of layers stacked in an anti-parallel manner parallel to (001). The hydro\u00adphobic aliphatic isopropyl groups protrude from these layers.A new polymorph of -2-amino-3-methyl\u00adpenta\u00adnoic acid, l-isoleucine C6H13NO2, crystallizes in the monoclinic space group P21 with four independent mol\u00adecules in the asymmetric unit. The mol\u00adecules are zwitterions. In the crystal, N\u2014H\u22efO hydrogen bonds link two pairs of independent mol\u00adecules and their symmetry-related counterparts to form two types of layers stacked in an anti-parallel manner parallel to (001). The hydro\u00adphobic aliphatic isopropyl groups protrude from these layers.A new polymorph of (2 S,3S)-2-Amino-3-methyl\u00adpenta\u00adnoic acid, known as l-isoleucine (l-Ile), is one of the 20 amino acids common in animal proteins and required for normal functioning in humans. l-Ile is classified as a hydro\u00adphobic amino acid and is one of the two common amino acids that has a chiral side chain. l-Ile is essential for human muscle tissue recovery after exercise, along with Valine and Leucine.\u00b0, N1\u2014C2\u2014C3\u2014C4 = \u2212155.4\u2005(3)\u00b0 and N3\u2014C14\u2014C15\u2014C18 = 78.1\u2005(4)\u00b0, N3\u2014C14\u2014C15\u2014C16 = \u2212155.8\u2005(3)\u00b0. The other two mol\u00adecules are of conformer type II with the torsion angles N2\u2014C8\u2014C9\u2014C12 = 178.6\u2005(4)\u00b0, N2\u2014C8\u2014C9\u2014C10 = \u221256.9\u2005(5)\u00b0 and N4\u2014C20\u2014C21\u2014C24 = 179.1\u2005(4)\u00b0, N4\u2014C20\u2014C21\u2014C22 = \u221256.8\u2005(5)\u00b0. Furthermore, there is a minor conformational variance between all the four independent mol\u00adecules, as illus\u00adtrated by the torsion angles of the iso-propyl side chains: C6\u2014C3\u2014C4\u2014C5 = \u221256.6\u2005(5)\u00b0, C12\u2014C9\u2014C10\u2014C11 = \u221251.6\u2005(6)\u00b0, C18\u2014C15\u2014C16\u2014C17 = \u221258.9\u2005(5)\u00b0 and C24\u2014C21\u2014C22\u2014C23 = \u221253.2\u2005(6)\u00b0.The existence of another chiral center in the side chain allows for conformational differences. Each l-Ile is embedded inside the layers while the side chains are oriented away, creating a hydro\u00adphobic surface. However, this hydrogen-bonding network has directionality along the polar b axis and specifically parallel to (001) was dissolved in water by heating to 353\u2005K, with constant stirring until total dissolution. The hot solution was then filtered through cotton wool into glass crystallization dishes, which were covered with filter paper in order to allow slow evaporation, placed in a heating bath. Colorless crystal chunks, suitable for X-ray crystallographic analysis were obtained. The absolute configuration of the title compound is already known.Single crystals of Uiso(H) = 1.2Ueq(C) or 1.5Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018006126/lh5872sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018006126/lh5872Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018006126/lh5872Isup3.cmlSupporting information file. DOI: 1838774CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II di\u00adthio\u00adcarbazate complex features a square-planar trans-N2S2 donor set for the metal atom 2], features a trans-N2S2 donor set as a result of the CuII atom being located on a crystallographic centre of inversion and being coordinated by thiol\u00adate-S and imine-N atoms derived from two di\u00adthio\u00adcarbazate anions. The resulting geometry is distorted square-planar. In the crystal, \u03c0(chelate ring)\u2013\u03c0(fur\u00adyl) [inter-centroid separation = 3.6950\u2005(14)\u2005\u00c5 and angle of inclination = 5.33\u2005(13)\u00b0] and phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adactions sustain supra\u00admolecular layers lying parallel to (Cg(fur\u00adyl) contacts (Cu\u22efCg = 3.74\u2005\u00c5) were also found. Inter\u00adaction energy calculations suggest the contacts between mol\u00adecules are largely dispersive in nature.The title Cu The conformation about the endocylic imine bond is Z, as a result of chelation, whereas the exocyclic imine bond has an E conformation.The mol\u00adecular structure of (I)s Table\u00a01. The res2CS chelate rings. While the r.m.s. deviation for the five atoms is relatively small at 0.0453\u2005\u00c5, suggesting a near planar ring, a better description for the conformation is that of an envelope with the copper atom being the flap atom. In this description, the r.m.s. deviation of the S1, N1, N2 and N3 atoms of the ring is 0.0002\u2005\u00c5, with the Cu atom lying 0.199\u2005(3)\u2005\u00c5 out of the plane. The dihedral angle between the best plane through the chelate ring and the 2-furyl ring is 5.33\u2005(18)\u00b0 indicating an essentially co-planar relationship. By contrast, the dihedral between the chelate and phenyl rings is 86.75\u2005(7)\u00b0, indicative of an orthogonal relationship. Finally, the dihedral angle between the peripheral organic rings is 81.42\u2005(9)\u00b0.The bidentate mode of the coordination of the di\u00adthio\u00adcarbazate ligand leads to the formation of five-membered CuNet al., 2008II in (I)ca 10\u00b0 in (I)ca 5\u00b0, changes consistent with the reorganization of \u03c0-electron density from the C1\u2014S1 to C1\u2014N1 bonds in (I)The structure of the acid form of the anion in (I)a). The association between mol\u00adecules is of the type \u03c0(chelate ring)\u2013\u03c0(fur\u00adyl) whereby the inter-centroid Cg\u2013Cgi separation is 3.6950\u2005(14)\u2005\u00c5 with angle of inclination = 5.33\u2005(13)\u00b0; symmetry operation (i) x, \u22121\u00a0+\u00a0y, z. Such \u03c0\u2013\u03c0 inter\u00adactions between chelate rings and aromatic rings are well documented in the literature, especially for sterically unencumbered square-planar complexes and can impart significant energies of stabilization to the mol\u00adecular packing , Table\u00a02b). Details of the weak inter\u00admolecular contacts connecting layers are given in the analysis of the calculated Hirshfeld surfaces below.The most prominent feature of the mol\u00adecular packing is the formation of supra\u00admolecular layers lying parallel to a), and those delineated into H\u22efH, C\u22efH/H\u22efC, S\u22efH/H\u22efS and C\u22efC contacts are illustrated in Fig.\u00a06b)\u2013(e), respectively; the percentage contribution from all the identified inter\u00adatomic contacts to the Hirshfeld surface are summarized qu\u00adanti\u00adtatively in Table\u00a04The overall two-dimensional fingerprint plot, Fig.\u00a06de + di \u223c2.1\u2005\u00c5 in the fingerprint plot delineated into H\u22efH contacts in Fig.\u00a06b), represents the short inter-layer H\u22efH contact involving phenyl-H8 atoms, Table\u00a03de + di \u223c2.7\u2005\u00c5 in the respective delineated fingerprint plot of Fig.\u00a06c) and Table\u00a03d), the short inter\u00adatomic contact involving the S-benzyl atoms, Table\u00a03de + di < 3.0\u2005\u00c5, i.e. at the sum of van der Waals radii. The distribution of points in the fingerprint plot delineated into C\u22efC contacts, Fig.\u00a06e), forming triangular tip at de + di \u223c3.3\u2005\u00c5 is due to the presence of such short inter\u00adatomic contacts summarized in Table\u00a03The conical tip appearing at Crystal Explorer , polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep). The energies were obtained using the wave function calculated at the HF/STO-3G level theory. The strength and nature of the inter\u00admolecular inter\u00adactions are summarized qu\u00adanti\u00adtatively in Table\u00a05x, \u22121\u00a0+\u00a0y, z in Table\u00a05Cg(fur\u00adyl) = 3.74\u2005\u00c5], \u03c0(chelate)\u2013\u03c0(fur\u00adyl), C\u22efC and S\u22efH/H\u22efS inter\u00adactions. Among these inter\u00adactions, the short inter\u00adatomic S\u22efH/H\u22efS contact contributes to the electrostatic component while the others to the dispersion component of the energies. Even though the inter-centroid distance between symmetry-related phenyl (C3\u2013C8) rings are greater than 4.0\u2005\u00c5 and the inter\u00adatomic S\u22efH distance is greater than sum of their van der Waal radii , they possess greater inter\u00adaction energies compared to inter\u00admolecular phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl) and short inter\u00adatomic H\u22efH contacts, as summarized in Table\u00a05b-axis direction in Fig.\u00a07Eele, Edisp and Etot, respectively; the radius of the cylinder is proportional to the magnitude of inter\u00adaction energy. It is clearly evident from the energy frameworks shown in Fig.\u00a07Utilizing et al., 2016e.g. substituents carrying pyridyl or phenoxide, neutral mol\u00adecules only and non-solvated structures yielded 24 analogues to (I)trans-N2S2 square-planar geom\u00adetry as in (I)2)5Me]=NN=CC6H4OMe-4}2]2 arising from inter\u00admolecular Cu\u22efS inter\u00adactions between centrosymmetrically related trans-N2S2 square-planar geometries was synthesized following a procedure adapted from a previous report : 3089 (w) \u03bd(N\u2014H), 1609 (m) \u03bd(C=N), 1016 (s) \u03bd(N\u2014N), 763 (s), \u03bd(C=S).26H22CuN4O2S4: C, 50.84; H, 3.61; N, 9.12; Cu, 10.34. Found; C, 50.49; H, 3.45; N, 8.77; Cu, 10.81. FTIR : 1593 (m), \u03bd(C=N), 964 (s), \u03bd(N\u2014N), 760 (s), \u03bd(C\u2014S).Synthesis of (I)Uiso(H) set to 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989019006145/hb7822sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019006145/hb7822Isup2.hklStructure factors: contains datablock(s) I. DOI: 1913482CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "We recently reported homologous [Ln(Cpttt)2][B(C6F5)4] (1\u2010Ln) for all the heavier Ln from Gd\u2013Lu; herein, we extend this motif to the early Ln. We find, for the largest LnIII cations, that contact ion pairs [Ln(Cpttt)2{(C6F5\u2010\u03ba1\u2010F)B(C6F5)3}] are isolated from reactions of parent [Ln(Cpttt)2(Cl)] (2\u2010Ln) with [H(SiEt3)2][B(C6F5)4], where the anion binds weakly to the equatorial sites of [Ln(Cpttt)2]+ through a single fluorine atom in the solid state. For smaller SmIII, [Sm(Cpttt)2][B(C6F5)4] (1\u2010Sm) is isolated, which like heavier 1\u2010Ln does not exhibit equatorial anion interactions, but the EuIII analogue 1\u2010Eu could not be synthesised due to the facile reduction of EuIII precursors to EuII products. Thus with the exception of Eu and radioactive Pm this work constitutes a structurally similar family of Ln metallocenium complexes, over 50 years after the [M(Cp)2]+ series was isolated for the 3d metals.As the dysprosocenium complex [Dy(Cp Herein, we extend these studies to the lighter, larger Ln to define the characteristic features of analogous [Ln(Cpttt)2]+ cations across the Ln series. We find that for the largest members of the 1\u2010Ln series one meta\u2010F of the [B(C6F5)4]\u2212 anion coordinates to the exposed equatorial site in the solid state, giving [Ln(Cpttt)2{(C6F5\u2010\u03ba1\u2010F)B(C6F5)3}] . However, smaller SmIII yields an isolated cation, [Sm(Cpttt)2][B(C6F5)4] (1\u2010Sm), which is structurally analogous to the heavier members of the series. We were unable to complete the series with the Eu analogue 1\u2010Eu as the stability associated with the +2 oxidation statettt)2(X)] precursors.We previously reported that the bulky ion [BC6F4]\u2212 provi1\u2010Ln ,ttt)2{(C6F5\u2010\u03ba1\u2010F)B(C6F5)3}] and [Sm(Cpttt)2][B(C6F5)4] (1\u2010Sm) were all synthesised by the reactions of [Ln(Cpttt)2(Cl)] with [H(SiEt3)2][B(C6F5)4]3)2][B(C6F5)4] was selected as a reagent for Ln metallocenium cation formation as: (i)\u2005it is soluble in benzene, hence strongly coordinating O\u2010donor solvents can be avoided; (ii)\u2005there is a significant enthalpic effect due to formation of a strong Si\u2212Cl bond, and (iii)\u2005there is an additional entropic effect arising from two reactant species giving three product species. Complexes 1\u2010Ln were crystallised from dichloromethane (DCM) layered with hexane and became more temperature\u2010sensitive as soon as DCM was added; thus, these complexes were stored at \u221225\u2009\u00b0C and measurements were performed below \u221225\u2009\u00b0C. Despite repeated efforts we were unable to recrystallise 1\u2010Ln from DCM/hexane; we later found that these complexes dissolved in hot toluene and could be recrystallised and stored at room temperature.According to previously published synthetic methods for the synthesis of 2\u2010Ln were synthesized from LnCl3 and two equivalents of KCpttt by modification of the synthesis of 2\u2010Pr and 2\u2010Nd from the parent LnCl3 and NaCpttt (Scheme\u20053 with NaCpttt gives [Eu(Cpttt)2], with elimination of half an equivalent of (Cpttt)2 via reductive coupling.ttt)2(F)] by the oxidation of [Eu(Cpttt)2]2][PF6]5 and [Fe(Cp)2]. However, this did not occur and instead we obtained [{Eu(Cpttt)(\u03bc\u2010PF6\u2010\u03ba4\u2010F)(THF)}2] (3) in 60\u2009% yield, along with several crystals of (Cpttt)2 (4), presumably due to the contamination of the EuII starting material with THF solvates, such as [Eu(Cpttt)2(THF)] or [Eu(Cpttt)(I)(THF)x]n, and subsequent ligand scrambling. Therefore, Eu is the only Ln apart from radioactive Pm that we were unable to access via these anion abstraction procedures. Crystalline yields for 2\u2010Ln are similar (46\u201352\u2009%) except for 2\u2010La, which was isolated in 31\u2009% yield. For 1\u2010Ln, the crystalline yields were dependent upon the solvent system; yields were significantly higher when crystallised from toluene (67\u2009% for 1\u2010La and 69\u2009% for 1\u2010Sm) compared to those crystallised from DCM layered with hexane (21\u201359\u2009%).The precursors t Scheme\u2005.19 This 1\u2010Ln and 2\u2010Ln were generally in excellent agreement with expected values, although low carbon values, likely due to carbide formation from incomplete combustion, were consistently obtained in some cases. In spite of this, the bulk purity of 1\u2010Ln was indicated by the consistency of other analytical data with their formulations (see below). Complex 1\u2010Sm contains the largest LnIII ion in a Ln metallocenium cation with no equatorial interactions to be isolated to date. The most appropriate explanation for the structural change for the largest LnIII cations is a simple argument based on ionic radii, with the two Cpttt rings offering a sufficient amount of steric protection around the smaller SmIII ion III ions are large enough to accommodate a further interaction .R ligands and the weakly coordinating anion.Elemental analysis results obtained for 1H\u2005NMR spectra were recorded from \u2212350 to +350\u2005ppm for 1\u2010Ln and 2\u2010Ln 4]\u2212 anions in 1\u2010Ln were observed by 11B{1H} and 19F{1H}\u2005NMR spectroscopy (see Supporting Information). In the case of 1\u2010La, we could also employ 13C{19F}\u2005NMR spectroscopy to characterise the [B(C6F5)4]\u2212 anion: the expected four aromatic resonances were located at 124.26 (ipso\u2010), 136.91 (m\u2010), 138.79 (p\u2010) and 148.71\u2005ppm (o\u2010); the signal for the ipso\u2010carbon is a quartet from coupling with 11B (1JBC=51.8\u2005Hz). Signals in the 11B{1H} NMR spectra of 1\u2010Ln were observed between \u221220 and \u221216\u2005ppm, and three peaks were observed in the 19F{1H}\u2005NMR spectra, corresponding to meta\u2010, para\u2010 and ortho\u2010F. In the 19F{1H}\u2005NMR spectrum of 1\u2010Sm, the para\u2010F signal is a triplet from coupling to two adjacent meta\u2010F atoms (JFF=20.4\u2005Hz), but this is a singlet for 1\u2010La, presumably due to broadening by quadrupolar 139La (I=7/2). The 11B and 19F\u2005NMR spectra of [Sc(Cp*)2{(C6F5\u2010\u03ba2\u2010F)B(C6F5)3}]2{(\u03bc\u2010C6F5\u2010\u03ba1\u2010F)2B(C6F5)2}]2 1\u2010Ln (Ln=Gd\u2013Lu).6F5)4]\u2212 anions in light 1\u2010Ln are only weakly bound in solution, and dissociate to give isolated [Ln(Cpttt)2]+ cations, as evidenced by the symmetry of 19F\u2005NMR spectra.19F\u2005NMR studies of 1\u2010La in C7D8, but the spectra were unchanged down to 218\u2005K. Finally, the paramagnetism of 3 precluded assignment of its 1H, 13C, 19F and 31P\u2005NMR spectra.1\u2010Ln, 2\u2010Ln, 3 and 4 were determined by single crystal XRD 2]+ cation of 1\u2010Sm exhibits identical structural features with 1\u2010Dy,meta\u2010fluorine of one of the C6F5 rings of the [B(C6F5)4]\u2212 anion. In all cases, the [Ln(Cpttt)2]+ fragments exhibit bent geometries and the Cpcentroid\u22c5\u22c5\u22c5Ln\u22c5\u22c5\u22c5Cpcentroid angles for 1\u2010Ln are closer to linearity than the corresponding precursors 2\u2010Ln. In the cases of 1\u2010Sm and the late Ln metallocenium cations in 1\u2010Ln ,centroid\u22c5\u22c5\u22c5Ln\u22c5\u22c5\u22c5Cpcentroid angles are significantly more linear than the [Ln(Cpttt)2]+ fragments in early 1\u2010Ln which have Ln\u22c5\u22c5\u22c5F interactions . Two nearly eclipsed \u03b75\u2010Cpttt rings coordinate to the LnIII centres in all 1\u2010Ln.centroid distances decrease regularly across the Ln series .1\u2010Sm, two equatorial electrostatic Ln\u22c5\u22c5\u22c5C interactions with carbon atoms from a tBu group were observed , with a C\u22c5\u22c5\u22c5Ln\u22c5\u22c5\u22c5C angle of 147.56(11)\u00b0; this feature has been noted for the heavier Lns previously.1\u2010Ln , Ln\u22c5\u22c5\u22c5F distances were observed between 2.679(9)\u2005\u00c5 (1\u2010La) to 2.632(4)\u2005\u00c5 (1\u2010Nd); similar contacts between LnCpR2 fragments and [B(C6F5)4]\u2212 anions have previously been seen for [Sc(Cp*)2{(C6F5\u2010\u03ba2\u2010F)B(C6F5)3}]2{(\u03bc\u2010C6F5\u2010\u03ba1\u2010F)2B(C6F5)2}]2 .2]+ fragments are bridged by two [B(C6F5)4]\u2212 anions to give a total of four meta\u2010F\u22c5\u22c5\u22c5Ln interactions, but in the Sc example a single C6F5 ring binds to one ScIII centre via both a meta\u2010 and a para\u2010F atom. The shortest Ln\u22c5\u22c5\u22c5F distance in 1\u2010Sm is 5.693(2)\u2005\u00c5, which indicates that Ln\u22c5\u22c5\u22c5F interactions are only observed for the 1\u2010Ln series when the LnIII ionic radius is larger than a critical value, when the bis\u2010Cpttt framework no longer provides sufficient coverage to prevent equatorial interactions with [B(C6F5)4]\u2212 anions.The solid\u2010state structures of to 149.48\u00b0; late L1\u2010Ln and 2\u2010Ln were obtained as microcrystalline powders , owing to the 4I9/2\u21924G5/2 transitions;1\u2010Sm shows absorptions at \u03bd\u02dcmax=11\u2009400\u2005cm\u22121 (\u03f5=20\u2005mol\u22121 dm3\u2005cm\u22121) and 8400\u2005cm\u22121 (\u03f5=50\u2005mol\u22121\u2009dm\u22123\u2009cm\u22121), which likely arise from 6H5/2\u21926F11/2, 6F7/2 transitions, respectively.2\u2010Nd and 2\u2010Sm.The electronic spectra of 1\u2010Ln and 2\u2010Ln using MOLCAS 8.0Complete active space self\u2010consistent field spin\u2010orbit (CASSCF\u2010SO) calculations were performed on unoptimized XRD structures of Jm projections of the total angular momentum J, but can also be defined as an effective S=1/2 state. With this definition, the S=1/2 states have an effective g\u2010matrix describing the magnetic anisotropy owing to the varying Jm composition of each doublet. The 3\u00d73 g\u2010matrix can always be diagonalised to yield three principal g\u2010values and their corresponding orientations in the molecular frame; here we define the principal g\u2010values such that g11/2), g1=g2=0 and g3=2JmJg where Jg is the Land\u00e9 g\u2010factor of the J multiplet of the LnIII ion. In the general case of mixed Jm states it is useful to define , the expectation value of the Jz operator which measures the projection of J on the quantisation axis, to understand the magnetic nature of the states; in the limiting case of a pure Jm state, =Jm.In the case of Kramers ions with an odd number of unpaired electrons, there will always be two\u2010fold electronic degeneracy in zero magnetic field, irrespective of a low\u2010symmetry CF. Each of these Kramers doublets is a linear combination of 1\u2010Ce , and is thus well\u2010described as Jm=\u00b15/2 when the CF is quantised along the g3 direction 4]\u2212 anion (Table\u2005S18), showing only a marginal increase in axiality. For 2\u2010Ce however, while the ground doublet is also Jm=\u00b15/2 with the CF defined along the Cpttt\u2013Cpttt direction , the first excited state is almost isotropic and the most excited state also resembles the Jm=\u00b15/2 state when the CF is quantised along the \u201cpseudo\u2010C3\u201d direction perpendicular to both the Cpttt\u2013Cpttt direction and the Ce\u2013Cl vector ; this demonstrates that the Cl\u2212 anion is competing with the Cpttt ligands and has a significant influence on the CF.For Ce Table\u2005 we obser1\u2010Sm , this makes 1\u2010Sm the antithesis of an SMM with the Jm=\u00b11/2, \u00b13/2 and \u00b15/2 doublets in ascending energy , yet when the CF is quantised along this direction there is strong mixing between Jm=\u00b11/2 and \u00b13/2 owing to the competition between the Cpttt and Cl\u2212 ligands (Table\u2005S29). For both 1\u2010Sm and 2\u2010Sm the g\u2010values are much reduced from those expected for an isolated 6H5/2 multiplet; this is due to considerable J\u2010mixing with the low\u2010lying 6H7/2 and 6H9/2 multiplets, rendering a CF model in the basis of a pure J=5/2 state an inaccurate representation of the wavefunction.The electronic structure of Sm Table\u2005, like 1\u2010y Figure\u2005 and S28.III the situation is less clear than for CeIII and SmIII because the magnitude of the quadrupolar terms in the expansion of the Jm electron densities are smaller than for the other LnIII ions,Jm states by the low symmetry CF is more severe. For 1\u2010Nd the two lowest lying doublets are 86\u2009% Jm=\u00b19/2 and 86\u2009% Jm=\u00b17/2, respectively, when the CF is quantised along the Cpttt\u2010Cpttt direction (Table\u2005S25), however the subsequent excited states are quite mixed and the highest\u2010energy doublet is 86\u2009% Jm=\u00b19/2 when the CF is quantised along the \u201cpseudo\u2010C3\u201d direction . Removal of the counterion reduces the axiality of the most excited state (Table\u2005S26), however this does not change the overall picture of the electronic structure. This suggests that it is the bent arrangement of the Cpttt rings, and not the influence of the [B(C6F5)4]\u2212 counterion, that forces the magnetic anisotropy of the most excited state to be perpendicular to that of the ground state, further highlighting the minimal electronic influence of the bulky anion. The electronic structure of 2\u2010Nd is generally similar to that of 1\u2010Nd, however the mixing is more severe (Table\u2005S27) and is not discussed further.For Ndpseudo\u2010doublets that are well described as pure Jm functions and a singlet state corresponding to Jm=0; the departure from axiality determines the degree to which the pseudo\u2010doublets are split into singlets. For pseudo\u2010doublets only a single principal g\u2010value can be defined and the other two are zero (note that this does not necessarily indicate a pure Jm state); thus, is a useful indicator for non\u2010Kramers ions.In the case of non\u2010Kramers ions, the action of a low\u2010symmetry CF can completely remove the degeneracy of the electronic states, although it does not have to do so (e.g. in high symmetry environments). In the case of a near\u2010axial CF, there will be a number of 1\u2010Pr . Removal of the counterion in the CASSCF\u2010SO calculations for 1\u2010Pr results in larger energy gaps between the CF states and reduces the splitting within the pseudo\u2010doublets at higher energies, but generally does not have a large effect on the electronic structure (Table\u2005S22). For 2\u2010Pr, with an extra competitive element in the CF (the Cl\u2212 ion), the resulting electronic structure is nearly all singlets (Table\u2005S23), and application of a small magnetic field is inconsequential (Table\u2005S24).The situation for Pr Table\u2005 is similn Tables\u2005 and S20,III, NdIII and SmIII) of 1\u2010Ln and 2\u2010Ln. We note generally that the intensities of the signals are very weak, however that there is excellent agreement between experiment and theory , however the latter is apparently too weak or is broadened beyond detection.To experimentally probe the nature of the ground states, we have performed cryogenic electron paramagnetic resonance (EPR) spectroscopy on the Kramers ion analogues , however the data for 2\u2010Pr decrease rapidly at lower temperatures, and those for 1\u2010Pr decrease slowly below 20\u2005K. For 2\u2010Pr this is due to a singlet ground state, which is confirmed by magnetisation measurements and CASSCF\u2010SO calculations (Table\u2005S23), but for 1\u2010Pr the experimental decrease is much more substantial than predicted by CASSCF\u2010SO, suggesting that the calculations have underestimated the splitting within the ground pseudo\u2010doublet on the order of a few cm\u22121 (Table\u2005S20). Generally, these magnetic data are in good agreement with CASSCF\u2010SO\u2010calculated electronic structures, however we note that due to the small magnetic moments of the light LnIII ions (where the SO coupling gives J=|L\u2212S| ground multiplets) the experimental data are sensitive to small errors in sample masses and diamagnetic corrections. The data for 1\u2010Sm and 2\u2010Sm are the most susceptible to this, as they feature not only the lowest magnetic moments, but also because they are highly sensitive to J mixing (see above) such that the calculations only reveal an approximate electronic structure.The \u03c7ttt)2]+ seriesIII analogues. Therefore, we investigated the Kramers ions with AC susceptibility studies to examine their magnetisation dynamics , giving an effective barrier of Ueff=51.2\u2005cm\u22121 with \u03c40=9.64\u00d710\u22128\u2005s; CASSCF\u2010SO predicts a first excited state at around 72\u2005cm\u22121, so this is not unreasonable. In all other cases a Raman mechanism is the best model for the data, and indeed is also a possible explanation for 2\u2010Nd. For both pairs of 1\u2010Ce vs. 2\u2010Ce and 1\u2010Nd vs. 2\u2010Nd, the Raman exponent is smaller for 1\u2010Ln than for 2\u2010Ln where the Cl\u2212 is bound, similar to the trend we have observed previously for the heavy Ln metallocenium cations (Table\u2005ttt)2]+ is responsible for lowered Raman exponents. However, the exponents for 1\u2010Ce and 1\u2010Nd are considerably larger than for 1\u2010Tb, 1\u2010Dy and 1\u2010Ho, which suggests that there could be an influence of the proximate [B(C6F5)4]\u2212 anion on the relaxation dynamics of these species.We were curious if the anomalous Raman exponents we observed previously for the heavy [Ln(Cpttt)2]+, with the exception of Eu and Pm. In the case of the early Ln (Ln=La\u2010Nd) the [B(C6F5)4]\u2212 counter\u2010ions weakly coordinate to the metal centre through a meta\u2010fluorine atom in the solid state, as the metals are large enough to incorporate this additional interaction; these equatorial interactions are absent in the solid state for the later Ln analogues which contain smaller Ln centres. However, in the solution phase these interactions could not be detected by VT 19F\u2005NMR spectroscopy, indicating that isolated Ln metallocenium cations are present in fluid solution.ttt)2]+ for the early Ln suggests that the weak equatorial meta\u2010fluorine interaction has little effect on the axiality of these systems. Measurement of the relaxation dynamics shows a consistent picture across the [Ln(Cpttt)2]+ series, where the absence of monodentate ligands leads to lower Raman exponents than when they are present, indicating that this effect is a hallmark of all Ln metallocenium cations. We cannot rule out the influence of [B(C6F5)4]\u2212 on the relaxation dynamics of 1\u2010Ce and 1\u2010Nd, and suggest that the effect of weak equatorial donors on magnetic relaxation mechanisms may be explored in the future by using different CpR ligands and a range of weakly coordinating anions.We have completed a series of analogous Ln metallocenium cations benzene and [D8]toluene were dried by refluxing over K, and [D2]DCM was dried by refluxing over CaH2. NMR solvents were degassed by three freeze\u2010pump\u2010thaw cycles, and vacuum\u2010transferred before use. Anhydrous LnCl3 were purchased from Alfa Aesar and were used as received. KCpttt,3)2][B(C6F5)4]ttt)2]2\u2010Ln were made by a modification of published procedures.1H (400 and 500\u2005MHz), 13C{1H} (100 and 125\u2005MHz), 13C{19F} (125\u2005MHz),11B{1H} (128 and 160\u2005MHz), and 19F{1H} (376\u2005MHz) NMR spectra were obtained on Avance III 400 or 500\u2005MHz spectrometers at 298\u2005K. UV/Vis/NIR spectroscopy was performed on samples in Youngs tap\u2010appended 10\u2005mm path length quartz cuvettes on an Agilent Technologies Cary Series UV/Vis/NIR spectrophotometer at 175\u20133300\u2005nm. ATR\u2010Fourier transform infrared (ATR\u2010FTIR) spectra were recorded as microcrystalline powders using a Bruker Tensor 27 spectrometer. Elemental analyses were performed by Mrs Anne Davies and Mr Martin Jennings at The University of Manchester School of Chemistry Microanalysis Service, Manchester, UK. General synthetic procedures for 1\u2010Ln and 2\u2010Ln are given below; full details are in the Supporting Information.ttt)2{(C6F5\u2010\u03ba1\u2010F)B(C6F5)3}] (La=La\u2010Nd), [Sm(Cpttt)2] [B(C6F5)4] (1\u2010Ln)[Ln(Cp: Benzene (15\u2005mL) was added to a mixture of [H(SiEt3)2][B(C6F5)4] (0.5\u20130.8\u2005mmol) and 2\u2010Ln (0.5\u20130.8\u2005mmol) at room temperature. The mixture was stirred for 16\u2005hours and a precipitate formed. The volatiles were removed in vacuo to give a powder, which was washed with hexane (10\u201315\u2005mL) and benzene (10\u201315\u2005mL). In some cases, the crude material was dissolved in DCM at \u221278\u2009\u00b0C, and layered with 1\u20131.5\u2005equiv of hexane. Storage at \u221225\u2009\u00b0C overnight gave crystals of 1\u2010Ln . In other cases, the crude material was dissolved in hot toluene (5\u2005mL). Storage at room temperature gave 1\u2010Ln .1\u2010La: Colourless crystals . Anal. Calcd (%) for C58H58BF20La: C, 54.22; H, 4.55; Found: C, 52.22; H, 4.15. 1H\u2005NMR : \u03b4=1.38 3), 1.46 3), 6.26\u2005ppm . 11B{1H}\u2005NMR : \u03b4=\u221216.63\u2005ppm (s). 13C{1H}\u2005NMR : \u03b4=29.86 and 30.00 (C(CH3)3), 30.53 (C(CH3)3), 31.03 and 31.18 (C(CH3)3), 32.82 (C(CH3)3), 135.96 (Cp\u2010CH), 147.78 (Cp\u2010C), 149.70\u2005ppm (Cp\u2010C). 13C{19F}\u2005NMR : \u03b4=124.26 , 136.91 , 138.79 , 148.71\u2005ppm . 19F{1H}\u2005NMR : \u03b4=\u2212166.98 , \u2212162.89 , \u2212132.67\u2005ppm . The low solubility of 1\u2010La in [D6]benzene precluded assignment of 1H and 13C{1H}\u2005NMR spectra in this solvent. 11B{1H} NMR : \u03b4=\u221216.00\u2005ppm (s). 19F{1H}\u2005NMR : \u03b4=\u2212165.32 , \u2212160.90 , \u2212131.66\u2005ppm . FTIR : \u03bd\u02dc=2967 , 2874 (w), 2819 (w), 1643 (m), 1513 (s), 1459 (s), 1365 (m), 1273 (m), 1240 (m), 1087 (s), 977 (s), 924 (w), 828 (s), 773 (s), 756 (s), 683 (s), 660 (s), 609 (m), 574 (m), 470 (w), 440\u2005cm\u22121 (w).1\u2010Ce: Yellow crystals . Anal. Calcd (%) for C58H58BF20Ce\u22c51.5CH2Cl2: C, 50.56; H, 4.35; Found: C, 50.90; H, 4.18. \u03bceff : 2.12\u2005\u03bcB. 1H\u2005NMR : \u03b4=\u221213.26 3), \u22127.93\u2005ppm 3); no other signals observed. 11B{1H}\u2005NMR : \u03b4=\u221218.02\u2005ppm (s). The paramagnetism of 1\u2010Ce precluded assignment of its 13C{1H}\u2005NMR spectrum. 19F{1H}\u2005NMR : \u03b4=\u2212170.26 , \u2212164.33 , \u2212134.61\u2005ppm . FTIR : \u03bd\u02dc=2963 , 2871 (w), 2821 (w), 1643 (m), 1512 (s), 1459 (s), 1365 (m), 1273 (w), 1241 (w), 1087 (s), 976 (s), 924 (w), 867 (w), 830 (m), 773 (s), 756 (s), 683 (s), 660 (s), 609 (m), 574 (m), 441\u2005cm\u22121 (w).1\u2010Pr: Yellow crystals . Anal. Calcd (%) for C58H58BF20Pr\u22c52\u2009CH2Cl2: C, 49.47; H, 4.29; Found: C, 47.68; H, 4.27. \u03bceff : 2.70\u2005\u03bcB. The paramagnetism of 1\u2010Pr precluded assignment of its 1H and 13C{1H}\u2005NMR spectra. 11B{1H}\u2005NMR : \u03b4=\u221219.42\u2005ppm (s). 19F{1H}\u2005NMR : \u03b4=\u2212172.86 , \u2212166.16 , \u2212136.51\u2005ppm . FTIR : \u03bd\u02dc=2963 (m), 2909 (w), 2871 (w), 1643 (m), 1460 (s), 1365 (m), 1261 (s), 1242 (w), 1086 (s), 1021 (m), 977 (s), 798 (s), 774 (m), 755 (m), 683 (s), 660 (s), 609 (w), 573 (m), 473 (w), 441\u2005cm\u22121 (w).1\u2010Nd: Green crystals . Anal. Calcd (%) for C58H58BF20Nd\u22c52\u2009CH2Cl2: C, 49.36; H, 4.28; Found: C, 49.51; H, 4.17. \u03bceff : 3.42\u2005\u03bcB. 1H\u2005NMR : \u03b4=\u221217.88 3), \u221211.94\u2005ppm 3); no other signals observed. 11B{1H} NMR : \u03b4=\u221218.55\u2005ppm (s). The paramagnetism of 1\u2010Nd precluded assignment of its 13C{1H}\u2005NMR spectrum. 19F{1H}\u2005NMR : \u03b4=\u2212170.31 , \u2212164.92 , \u2212135.17\u2005ppm . FTIR : \u03bd\u02dc=2967 , 2875 (w), 2820 (w), 1643 (m), 1512 (s), 1460 (s), 1394 (w), 1365 (m), 1254 (w), 1241 (w), 1087 (s), 977 (s), 955 (w), 924 (w), 829 (m), 773 (s), 756 (s), 683 (s), 660 (s), 609 (w), 574 (m), 477 (w), 440\u2005cm\u22121 (w).1\u2010Sm: Red crystals . Anal. Calcd (%) for C58H58BF20Sm: C, 53.74; H, 4.51; Found: C, 53.88; H, 4.49. \u03bceff : 1.90\u2005\u03bcB. The paramagnetism of 1\u2010Sm precluded assignment of its 13C{1H} NMR spectrum. 1H\u2005NMR : \u03b4=\u22121.37 3), 19.69\u2005ppm ; no other signals observed. 11B{1H}\u2005NMR : \u03b4=\u221216.76\u2005ppm (s). 19F{1H}\u2005NMR : \u03b4=\u2212167.89 , \u2212163.76 , \u2212133.13\u2005ppm . 1H\u2005NMR : \u03b4=\u22122.58 3), \u22120.90 3), 18.80\u2005ppm . 11B{1H}\u2005NMR : \u03b4=\u221216.32\u2005ppm (s). 19F{1H}\u2005NMR : \u03b4=\u2212168.96 , \u2212162.02 , \u2212131.93\u2005ppm . FTIR : \u03bd\u02dc=2964 , 2872 (w), 2796 (w), 1642 (m), 1512 (s), 1459 (s), 1367 (m), 1276 (m), 1239 (m), 1083 (s), 977 (s), 843 (w), 806 (w), 774 (m), 756 (m), 735 (m), 683 (m), 661 (s), 611 (w), 574 (w), 466 (w), 449\u2005cm\u22121 (w).ttt)2(Cl)] (2\u2010Ln)[Ln(Cp: THF (30\u2005mL) was added to a Teflon tap\u2010appended ampoule containing a pre\u2010cooled (\u221278\u2009\u00b0C) mixture of LnCl3 (2\u2005mmol) and KCpttt (4\u2005mmol). The reaction mixture was allowed to warm to room temperature and heated in an oil bath at 80\u2009\u00b0C for 16\u2005hours. The solvent was removed in vacuo and toluene (30\u2005mL) was added. The reaction mixture was heated in an oil bath at 120\u2009\u00b0C for 16\u2005hours. The resultant suspension was allowed to settle for 3\u2005hours and filtered. The solution was concentrated to 2\u2005mL and stored at 8\u2009\u00b0C to afford crystals of 2\u2010Ln.2\u2010La: Colourless crystals . Anal. Calcd (%) for C34H58LaCl: C, 63.69; H, 9.12; Found: C, 63.57; H, 9.30. 1H\u2005NMR : \u03b4=1.25 3), 1.52 3), 6.51\u2005ppm . 13C{1H}\u2005NMR : \u03b4=31.19 (C(CH3)3), 32.86 (C(CH3)3), 34.50 (C(CH3)3), 34.68 (C(CH3)3), 114.78 (Cp\u2010CH), 138.17 (Cp\u2010C), 139.58\u2005ppm (Cp\u2010C). FTIR : \u03bd\u02dc=2956 (s), 2904 (w), 2869 (w), 1461 (m), 1389 (m), 1361 (s), 1260 (s), 1241 (m), 1091 , 1016 (s), 866 (w), 797 (s), 678 (s), 590 (w), 566 (w), 551 (w), 436\u2005cm\u22121 (w).2\u2010Ce: Orange crystals . Anal. Calcd (%) for C34H58CeCl: C, 63.57; H, 9.10. Found: C, 63.60; H, 9.22. \u03bceff : 2.34\u2005\u03bcB. The paramagnetism of 2\u2010Ce precluded assignment of its 13C{1H} NMR spectrum. 1H\u2005NMR : \u03b4=\u221213.06 3), \u22122.53\u2005ppm 3); no other signals observed. FTIR : \u03bd\u02dc=2954 (s), 2904 (m), 2868 (w), 1460 (s), 1389 (s), 1358 (s), 1241 (s), 1165 (m), 1001 (s), 958 (m), 816 (s), 774 (s), 678 (s), 566 (w), 436\u2005cm\u22121 (s).2\u2010Pr: Pale green crystals . Anal. Calcd (%) for C34H58PrCl: C, 63.49; H, 9.09; Found: C, 63.37; H, 9.22. \u03bceff : 3.35\u2005\u03bcB. The paramagnetism of 2\u2010Pr precluded assignment of its 13C{1H}\u2005NMR spectrum. 1H\u2005NMR : \u03b4=\u221236.08 3), \u22127.74\u2005ppm 3); no other signals observed. FTIR : \u03bd\u02dc=2955 (s), 2905 (m), 2869 (w), 1460 (s), 1389 (s), 1359 (s), 1241 (s), 1166 (m), 1001 (s), 959 (m), 832 (m), 818 (s), 775 (s), 679 (s), 567 (m), 437\u2005cm\u22121 (s).2\u2010Nd: Blue crystals . Anal. Calcd (%) for C34H58NdCl: C, 63.16; H, 9.04; Found: C, 61.22; H, 9.02. The paramagnetism of 2\u2010Nd precluded assignment of its 13C{1H}\u2005NMR spectrum. \u03bceff : 3.55\u2005\u03bcB. 1H\u2005NMR : \u03b4=\u221218.95 3), \u22125.58\u2005ppm 3); no other signals observed FTIR : \u03bd\u02dc=2955 (s), 2907 (m), 2870 (w), 1460 (s), 1389 (s), 1358 (s), 1241 (s), 1166 (m), 1001 (s), 833 (s), 820 (s), 775 (s), 679 (s), 439\u2005cm\u22121 (s).2\u2010Sm: Yellow crystals . Anal. Calcd (%) for C34H58SmCl: C, 62.57; H, 8.96; Found: C, 60.93; H, 9.19. \u03bceff : 1.69\u2005\u03bcB. The paramagnetism of 2\u2010Sm precluded assignment of its 13C{1H}\u2005NMR spectrum. 1H\u2005NMR : \u03b4=\u22126.01 3), 0.55 3), 19.80\u2005ppm . FTIR : \u03bd\u02dc=2956 (s), 2905 (m), 2870 (w), 1460 (s), 1389 (s), 1356 (s), 1241 (s), 1221 (w), 1166 (s), 1000 (s), 959 (m), 836 (w), 824 (s), 776 (s), 679 (s), 591 (w), 438\u2005cm\u22121 (s).The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "Scientific Reports 10.1038/s41598-017-01160-1, published online 21 April 2017Correction to: This Article contains errors in the first column of Table\u00a0In addition, Table \u201coral 200\u2009mg preoperative\u201dshould read:\u201coral 200\u2009\u03bcg preoperative\u201d"} +{"text": "In salt(II), the anion consists of a 4-meth\u00adoxy\u00adbenzene\u00adsulfonate ion, and it crystallizes as a monohydrate.The title mol\u00adecular salts are stilbazole, or 4-styryl\u00adpyridine, derivatives in which the cation has a methyl group attached to the pyridine ring N atom and a diethyl amine group attached to the benzene ring. In salt (I), the cadmium atom of the [CdI 18H23N2)2[CdI4], (I), and C18H23N2+\u00b7C7H7O4S\u2212\u00b7H2O, (II), are stilbazole, or 4-styryl\u00adpyridine, derivatives. The cation, (E)-4-[4-(di\u00adethyl\u00adamino)\u00adstyr\u00adyl]-1-methyl\u00adpyridin-1-ium, has a methyl group attached to pyridine ring and a diethyl amine group attached to the benzene ring. The asymmetric unit of salt (I), comprises one cationic mol\u00adecule and half a CdI4 dianion. The Cd atom is situated on a twofold rotation axis and has a slightly distorted tetra\u00adhedral coordination sphere. In (II), the anion consists of a 4-meth\u00adoxy\u00adbenzene\u00adsulfonate and it crystallizes as a monohydrate. In both salts, the cations adopt an E configuration with respect to the C=C bond and the pyridine and benzene rings are inclined to each other by 10.7\u2005(4)\u00b0 in (I) and 4.6\u2005(2)\u00b0 in (II). In the crystals of both salts, the packing is consolidated by offset \u03c0\u2013\u03c0 stacking inter\u00adactions involving the pyridinium and benzene rings, with centroid\u2013centroid distances of 3.627\u2005(4)\u2005\u00c5 in (I) and 3.614\u2005(3)\u2005\u00c5 in (II). In the crystal of (II), a pair of 4-meth\u00adoxy\u00adbenzene\u00adsulfonate anions are bridged by Owater\u2014H\u22efOsulfonate hydrogen bonds, forming loops with an R24(8) motif. These four-membered units are then linked to the cations by a number of C\u2014H\u22efO hydrogen bonds, forming slabs lying parallel to the ab plane.The title mol\u00adecular salts, (C This shows that both compounds exhibit very good anti\u00adcancer activity, which implies that they may be suitable for biomedical applications.Stilbene-based compounds have been reported to possess a wide range of biological applications including anti\u00adbacterial (Chanawanno E)-4-[4-(di\u00adethyl\u00adamino)\u00adstyr\u00adyl]-1-methyl\u00adpyridin-1-ium. Their mol\u00adecular structures are illustrated in Fig.\u00a014]2\u2212 anion in the asymmetric unit, the cadmium atom being located on a twofold rotation axis. The cadmium atom is surrounded by four iodine atoms with a slightly distorted tetra\u00adhedral coordination sphere. In salt (II)E, with the C4\u2014C7=C8\u2014C9 torsion angle being 179.6\u2005(6) \u00b0 in (I)The title mol\u00adecular salts consist of the same cationic stilbazole derivative, and benzene (C9\u2013C14) rings are 10.7\u2005(4) and 4.6\u2005(2)\u00b0 in (I)Cg2) and pyridine rings .In the crystal of (I)water\u2014H\u22efOsulfonate hydrogen bonds, forming loops with an ab plane (Table\u00a01Cg2\u22efCg1ii = 3.614\u2005(3)\u2005\u00c5, \u03b1 = 4.6\u2005(2)\u00b0, \u03b2 = 15.5\u00b0, inter\u00adplanar distances are 3.425\u2005(2) and 3.484\u2005(2)\u2005\u00c5, offset = 0.963\u2005\u00c5, symmetry code: (ii) x\u00a0\u2212\u00a01, y, z].In the crystal of (II)f Table\u00a01. These fe Table\u00a01. Within et al., 2016et al., 2004ca 4.08\u00b0, while in the tetra\u00adphenyl\u00adborate salt A search of the Cambridge Structural Database \u00b0 in salt (II)p-toluene\u00adsulfonate anions via O\u2014H\u22efO hydrogen bonds, forming an There is only one salt reported with the title cation and a sulfonate anion, namely the Compound (I)E)-4-[4-(di\u00adethyl\u00adamino)\u00adstyr\u00adyl]-1-methyl-pyridinium-iodide and cadmium iodide were dissolved in a composite solvent, 2:1 ratio of aceto\u00adnitrile and double-distilled water. The mixture was stirred well at 343\u2005K and then allowed to cool naturally to room temperature. The solution was filtered and the filtrate left for the solvent to slowly evaporate at room temperature. After 3\u20134 weeks, dark-brown block-like crystals of compound (I)(Compound (II)E)-4-[4-(di\u00adethyl\u00adamino)\u00adstyr\u00adyl]-1-methyl\u00adpyridinium iodide was mixed with sodium 4-meth\u00adoxy\u00adbenzene\u00adsulfonate in distilled water and heated at 373\u2005K for 30\u2005min. The mixture immediately yielded a grey precipitate of sodium iodide. After stirring the mixture for 30\u2005min, the sodium iodide precipitate was removed. The filtrate was left to slowly evaporate and gave a deep-red solid. Red block-like crystals of compound (II)(Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq for other H atoms. The rotation angles for the methyl groups were optimized by least-squares. In compound (II)d(O\u2014H) = 0.85\u2005\u00c5 and Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details for salts (I)10.1107/S2056989018016808/su5462sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989018016808/su5462Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989018016808/su5462IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989018016808/su5462IIsup4.cmlSupporting information file. DOI: 1589674, 1589675CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-(pyridin-2-ylmeth\u00adyl)benzamide derivative, crystallizes with two independent mol\u00adecules (A and B) in the asymmetric unit, which differ essentially in the orientation of the pyridine ring with respect to the benzene ring, with the two rings being inclined to each other by 53.3\u2005(2) and 72.9\u2005(2)\u00b0 in mol\u00adecules A and B, respectively.The title 18H18F2N2O3, crystallizes with two independent mol\u00adecules (A and B) in the asymmetric unit. They differ essentially in the orientation of the pyridine ring with respect to the benzene ring; these two rings are inclined to each other by 53.3\u2005(2)\u00b0 in mol\u00adecule A and by 72.9\u2005(2)\u00b0 in mol\u00adecule B. The 3-(cyclo\u00adpropyl\u00admeth\u00adoxy) side chain has an extended conformation in both mol\u00adecules. The two mol\u00adecules are linked by a pair of C\u2014H\u22efO hydrogen bonds and two C\u2014H\u22ef\u03c0 inter\u00adactions, forming an A\u2013B unit. In the crystal, this unit is linked by N\u2014H\u22efO hydrogen bonds, forming a zigzag \u2013A\u2013B\u2013A\u2013B\u2013 chain along [001]. The chains are linked by C\u2014H\u22efN and C\u2014H\u22efF hydrogen bonds to form layers parallel to the ac plane. Finally, the layers are linked by a third C\u2014H\u22ef\u03c0 inter\u00adaction, forming a three-dimensional structure. The major contributions to the Hirshfeld surface are those due to H\u22efH contacts (39.7%), followed by F\u22efH/H\u22efF contacts (19.2%).The title compound, C The dihedral angle between the benzamide ring and the pyridine ring is 53.3\u2005(2)\u00b0 in mol\u00adecule A and 72.9\u2005(2)\u00b0 in mol\u00adecule B. The cyclo\u00adpropane ring makes a dihedral angle of 57.7\u2005(5)\u00b0 with the benzene ring in mol\u00adecule A and 58.7\u2005(4)\u00b0 in mol\u00adecule B. The sum of the bond angles around atom N1 (359.9\u00b0) is in accordance with sp2 hybridization in both mol\u00adecules. The bond lengths and bond angles in both mol\u00adecules are comparable with those reported for a similar compound, 3-(cyclo\u00adpropyl\u00admeth\u00adoxy)-N--4-(di\u00adfluoro\u00admeth\u00adoxy) benzamide -4-(di\u00adfluoro\u00admeth\u00adoxy) benzamide are very similar. Considering the low-temperature phase PEDWOM only, in each mol\u00adecule the benzene and pyridine rings are positioned almost perpendicular to each other, with dihedral angles of 88.38\u2005(14), 89.34\u2005(14) and 84.72\u2005(14)\u00b0, compared to 53.3\u2005(2) and 72.9\u2005(2)\u00b0 for mol\u00adecules A and B, respectively, in the title compound. In PEDWOM the cyclo\u00adpropane ring makes dihedral angles of 55.43\u2005(3), 49.6\u2005(3) and 50.9\u2005(3)\u00b0 with the corresponding benzene ring. These dihedral angles are very similar to those observed in the title structure [57.7\u2005(5)\u00b0 in mol\u00adecule A and 58.7\u2005(4)\u00b0 in mol\u00adecule B]. In the second compound, methyl 3-(cyclo\u00adpropyl\u00admeth\u00adoxy)-4-hy\u00addroxy\u00adbenzoate , followed by F\u22efH/ H\u22efF at 19.2% , C\u22efH/H\u22efC at 16.6% , O\u22efH/ H\u22efO at 14.0% , N\u22efH/H\u22efN at 6.8% . Hence, the H\u22efH and F\u22efH/H\u22efF inter\u00admolecular contacts are the most abundant in the crystal packing, and make the most significant contributions to the total Hirshfeld surfaces.The Hirshfeld surface analysis in methanol were heated first to 393\u2005K for 2\u2005h in the presence of the inexpensive ionic liquid tetra\u00adbutyl\u00adammonium bromide (TBAB). The reaction was monitored by TLC, and on completion the reaction mixture was allowed to cool to room temperature, then filtered to remove the insoluble solids. The filtered solid was then washed with di\u00adchloro\u00admethane. Excess solvents were removed under reduced pressure and the obtained crude product was purified by crystallization using 1:1 ratio of chloro\u00adform and methanol. Colourless block-like crystals were obtained after two days.A mixture of 4-(di\u00adfluoro\u00admeth\u00adyl)-3-hy\u00addroxy\u00adbenzoic acid (2\u2005mmol), (chloro\u00admeth\u00adyl)cyclo\u00adpropane (2\u2005mmol) and 2-picolyl\u00adamine (3\u2005mmol) with PPhUiso(H) =1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. The absolute structure of the mol\u00adecules in the crystal are unknown; the Flack parameter refined to 0.6\u2005(3).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019012866/su5514sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019012866/su5514Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019012866/su5514Isup3.cmlSupporting information file. DOI: 1954107CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Though such interactions are commonly attributed to \u201csigma-hole\u201d-type electrostatic effects, we show that they exhibit profound similarities and analogies to the resonance-type 3-center, 4-electron (3c/4e) donor-acceptor interactions of hydrogen bonding, where classical-type \u201celectrostatics\u201d are known to play only a secondary modulating role. The general 3c/4e resonance perspective corresponds to a continuous range of interatomic A\u00b7\u00b7\u00b7B bond orders (bAB), spanning both the stronger \u201ccovalent\u201d interactions of the molecular domain and the weaker interactions that underlie supramolecular complexation phenomena. We show how a unified NBO/NRT-based description of hydrogen, halogen, pnicogen, and related bonding yields an improved predictive utility and intuitive understanding of empirical trends in binding energies, structural geometry, and other measurable properties that are expected to be manifested in all such supramolecular interaction phenomena.We employ a variety of natural bond orbital (NBO) and natural resonance theory (NRT) tools to comprehensively investigate the nature of halogen and pnicogen bonding interactions in RPH Recent computational ,2,3,4 annB\u2192\u03c3*AH donor\u2013acceptor interaction between the lone pair (nB) of the Lewis base B and the valence antibond (\u03c3*AH) of the Lewis acid A\u2014H. In natural resonance theory (NRT) [nB\u2192\u03c3*AH interaction represents resonance mixing between the parent natural Lewis structure (NLS) I and the secondary \u201ccharge\u2013transfer\u201d resonance structure II (with respective NRT weightings wI and wII):bAH, bBH bond orders are necessarily related by the characteristic \u201cbond conservation\u201d relationship [wI + wII = 1) for the composite resonance hybrid. Even in the absence of NRT bond order descriptors, the importance of NBO nB\u2192\u03c3*AH delocalization can be assessed by 2nd-order perturbative energy (\u0394E(2)n\u03c3*), intermolecular charge-transfer (Qn\u03c3*), or the deletion of this NBO interaction (\u0394E$DELn\u03c3*), as well as the variational recalculation of structure, energetics, and vibrational frequencies [In the case of hydrogen bonding, the primacy of short-range covalency forces is now widely recognized ,36,37,38ry (NRT) ,41,42,43rchanged . As recorchanged , the resrchanged ,47. In ttionship ,bAH + bquencies . In contquencies that mos2\u00b7\u00b7\u00b7IF/FI that exhibit a wide range of binding strengths. A smaller subset of such species was previously examined [In the present work, we take up the analogous covalency versus electrostatics questions for title complexes RPHexamined with an examined .n-\u03c3* interaction) is strictly absent in uncorrelated Hartree\u2013Fock theory. Though an empirical dispersion correction [3PH2\u00b7\u00b7\u00b7IF (see Ebind). Such \u201ccorrections\u201d are not considered further in the present work. Note that we ignore the weaker effects of the London dispersion interaction , a pure rrection is oftenE(2)n\u03c3*, charge\u2013transfer Qn\u03c3*, and $DEL-deletion \u0394E$DELn\u03c3* measures of donor\u2013acceptor attraction [E(pw)n\u03c3) of filled orbitals [In the following, we first describe the species, computational levels, and NBO/NRT methods to be employed. The latter include NBO \u0394traction , the cororbitals that comorbitals . We then2\u00b7\u00b7\u00b7XY that included all possible dihalogen species of the first four periodic rows. However, from a qualitative conceptual perspective, it soon became apparent that the properties of such complexes vary in a smooth and chemically reasonable fashion with dihalogen polarity, as maximized in the iodine\u2013fluorine (IF) species. Accordingly, we focus here on the \u201cpolar extremes\u201d of RPH2\u00b7\u00b7\u00b7IF versus RPH2\u00b7\u00b7\u00b7FI complexation, corresponding to opposed signs of any envisioned \u201cdipole\u2013dipole\u201d contributions to intermolecular binding. A direct comparison of these two extremes thereby allows one to recognize the important modulating influence of classical dipole\u2013dipole forces , while also verifying that resonance-type covalency forces yield robust supramolecular bonding even when the presumed classical electrostatic prerequisites for such complexation are profoundly violated. While the computational results presented here focus only on the extremal RPH2\u00b7\u00b7\u00b7IF versus RPH2\u00b7\u00b7\u00b7FI limits of this broader picture, the Our studies began with a larger data set of dihalogen complexes RPH2 monomers and LANL2DZ for dihalogen monomers of each complex. We employ simple B3LYP density functional methodology throughout this work, but alternative MP2 evaluations at different basis levels were also performed for comparison (see SI). All structures were optimized, and frequencies were computed to confirm equilibrium geometries corresponding to minimum energy points. The binding energy, \u0394Ebind = E(AB) \u2212 E(A) \u2212 E(B), was calculated as the direct energy difference between optimized dimer and relaxed monomers. All electronic structure calculations were performed with the Gaussian 16 program package [NBO 7.0 software [NBOPro@Jmol program [Though the conceptual picture we describe is insensitive to many details of the chosen computational method and basis set , the inc package . NBO/NRTsoftware , and bot program .2\u00b7\u00b7\u00b7IF and RPH2\u00b7\u00b7\u00b7FI species as obtained at the adopted \u201cB3LYP/mixed-PP\u201d level, showing the wide range of tuning by various R-substituents. Despite the fact that all complexes are appreciably bound , interesting structural contrasts are immediately evident between these two classes of species. Focusing first on the orientation angles (\u0398) in 2\u00b7\u00b7\u00b7IF species all maintain nearly linear P\u00b7\u00b7\u00b7IF alignment (\u0398PIF \u2248 180\u00b0) and a circa 120\u00b0 bend angle between RP and IF axes (\u0398RPI \u2248 120\u00b0). In contrast, the RPH2\u00b7\u00b7\u00b7FI species are noticeably tilted away from P\u00b7\u00b7\u00b7FI linearity (\u0398PFI \u2248 160\u00b0), instead adopting increasingly near-linear alignment with respect to RP and FI axes (\u0398RPF \u2248 170\u00b0). Such structural tendencies suggest that the principal underlying \u201clinearizing\u201d (H-bond-like) interaction differs in the two cases and that other RPH2\u00b7\u00b7\u00b7XY dihalogen species of intermediate polarity will likewise exhibit intermediate orientational preferences between these two competing tendencies. Indeed, this is the broader picture obtained from the full set of optimized dihalogen structures, with the homopolar RPH2\u00b7\u00b7\u00b7X2 species forming the approximate \u201cdividing line\u201d between RPH2\u00b7\u00b7\u00b7IF-like versus RPH2\u00b7\u00b7\u00b7FI-like structural propensity. These structural features of RPH2\u00b7\u00b7\u00b7IF-like (\u201cP\u00b7\u00b7\u00b7IF\u201d) versus RPH2\u00b7\u00b7\u00b7FI-like (\u201cRP\u00b7\u00b7\u00b7F\u201d) complexes are also consistently exhibited at other theory levels, as shown in SI 2\u00b7\u00b7\u00b7IF versus RPH2\u00b7\u00b7\u00b7FI species for describing the full range of RPH2\u00b7\u00b7\u00b7XY complexation , including general consistency with previous MP2/aug-cc-pVTZ-level calculations.RP\u00b7\u00b7\u00b7F or RP\u00b7\u00b7\u00b7I with increasing binding energy \u0394Ebind is also discernable in the details of the figure panels. From both Ebind with various R-substituents for the strongly bound complexes of the RPH2\u00b7\u00b7\u00b7IF type. The graph includes comparison values for the alternative theory levels given in IF with respect to the R-substituent . As shown in the figure, \u0394\u03bdIF is strongly red-shifted for the most strongly bound complexes, similar to the well-known spectroscopic signature of H-bonding. IF between these experimentally observable quantities that confirms the similarity to H-bonding.ding. IF versus \u0394F to achieve the net binding energies displayed in Ebind binding affinity and RP\u00b7\u00b7\u00b7I separation in the deep sub-vdW region.Returning to the structural data of 2\u00b7\u00b7\u00b7IF and weak RPH2\u00b7\u00b7\u00b7FI complexes, the structural features hint at striking resemblances to familiar H-bonding phenomena, including deep sub-vdW penetration and pronounced \u201clinearization\u201d tendency . We take up NBO/NRT investigation of these apparent similarities and bonding relationships in the following sections.The computed structural and binding characteristics noted above identify the propensities and patterns to be explained and provide helpful clues to the nature of the underlying attractive forces. Even the more weakly bound complexes of the P\u00b7\u00b7\u00b7FI type exhibit optimized P\u00b7\u00b7\u00b7F distances (2.1\u20132.6\u00c5) that lie significantly (>0.5\u00c5) inside vdW contact donor\u2013acceptor interactions of the present species.As suggested in above, an NBO/NRT analysis of B:\u00b7\u00b7\u00b7HA hydrogen-bonding phenomena reveals a host of characteristic correlations ,23,60 thanalysis ,62,63. Tanalysis ,66,67,682\u00b7\u00b7\u00b7IF complexes, the key feature of envisioned P\u00b7\u00b7\u00b7I\u2014F \u201chalogen bonding\u201d is the donor\u2013acceptor delocalization of electronic charge from an occupied donor NBO of P (lone pair nP) to the favorably polarized acceptor NBO (antibond \u03c3*IF) of the P\u00b7\u00b7\u00b7I\u2014F triad. For RPH2\u00b7\u00b7\u00b7FI complexes, where the nP-\u03c3*IF overlap is unfavorable, the analogous feature of envisioned F\u00b7\u00b7\u00b7P\u2014R \u201cpnicogen bonding\u201d is the donation from a fluorine lone pair (nF) to the proximal antibond (\u03c3*PR) of the phosphine monomer. Each such n\u2192\u03c3* delocalization can be alternatively quantified in terms of energetic (perturbative \u0394E(2)n\u03c3* or variational deletion \u0394E($DEL)n\u03c3*), charge transfer (Qn\u03c3*), or fractional NRT bond order (bP\u00b7\u00b7\u00b7I or bP\u00b7\u00b7\u00b7F) descriptors.The analogies of the preceding section suggest appropriate NBO/NRT descriptors for the corresponding \u00b7\u00b7\u00b7X\u2014 (\u201cX-ogen\u201d) species that can be similarly visualized in simple orbital overlap terms. For RPHEn\u03c3* stabilization energy associated with each type of delocalization in the strongly bound RPH2\u00b7\u00b7\u00b7IF species. The first two columns display 2nd-order perturbative \u0394E(2)n\u03c3* estimates for nP\u2192\u03c3*IF and nI\u2192\u03c3*PR (\u201cpnicogen bond\u201d) contributions. The final two columns of E($DEL)n\u03c3* variational deletion estimates, obtained as the variational energy raising when the specific n\u2192\u03c3* NBO interaction is deleted from the total energy evaluation. The two estimates are seen to be in reasonable qualitative agreement for each interaction type. The \u0394E(2)n(P)\u2192\u03c3*(IF) values range from 19 to 37 kcal/mol (in the same order as \u0394Ebind), whereas \u0394E(2)n(I)\u2192\u03c3*(FI) is relatively negligible in each case, and a similar pattern is seen in the \u0394E($DEL)n\u03c3* values. We therefore expect that the stabilization energy of the nP\u2192\u03c3*IF interaction is the dominant attractive contribution to the structure and binding of RPH2\u00b7\u00b7\u00b7IF complexes, similar to the dominance of the nB\u2192\u03c3*AH interaction in H-bonding.2\u00b7\u00b7\u00b7FI species, the situation is more complex. Due to the unfavorable polarization of \u03c3*FI away from the nP lone pair, the single nP\u2192\u03c3*IF interaction that dominates bonding in RPH2\u00b7\u00b7\u00b7IF species becomes only one of several such competing contributions to net binding. Specifically, for the opposed F\u00b7\u00b7\u00b7P\u2014R (pnicogen bonding) motif, three possible nF lone pairs (one of sigma type and two of pi type) and three \u03c3*PY acceptors (one \u03c3*RP and two \u03c3*PH) are within proximal interaction range as potential contributors to a complex resonance mixture. A particular interaction such as nF(\u03c3)\u2192\u03c3*RP therefore represents only one of the nine related n\u2192\u03c3* interactions that may exert leverage on angular structure and binding energy. In such a case, only a more nuanced resonance-type description (see the discussion of NRT bond orders below) can describe the complete bonding picture that provides useful correlations with experimentally measurable properties. We therefore defer further discussion of RPH2\u00b7\u00b7\u00b7FI complexes to a later subsection.For the weakly bound RPHn\u2192\u03c3* interactions of halogen or pnicogen (nI(\u03c0)\u2192\u03c3*RP, nI(\u03c3)\u2192\u03c3*RP; final two columns) types, with corresponding numerical \u0394E(2)n\u03c3* values is much stronger than the \u201cedge-on\u201d (pi-type) overlap of \u03c3*RP with the in-plane iodine nI(\u03c0) (middle panel) or nI(\u03c3) (right panel) lone pairs, in accordance with the \u0394E(2)n\u03c3* values shown in each panel. Even a casual glance at the overall patterns of the orbital overlap in nP\u2192\u03c3*IF) rather than pnicogen-type (nI(\u03c0)\u2192\u03c3*RP, nI(\u03c3)\u2192\u03c3*RP) bonding throughout the RPH2\u00b7\u00b7\u00b7IF series, as numerically displayed in For example, in the three panels for CHEn\u03c3* stabilization energies of Ebind binding energies of En\u03c3 donor\u2013donor interaction. E(pw)n-\u03c3 for repulsive \u201csteric exchange energy\u201d [2\u00b7\u00b7\u00b7IF series. As shown in the panels, the two leading intermolecular repulsions refer to the nP-nI steric clash between lone pairs of proximal P and I atoms (left) and nP-\u03c3IF clash between a lone pair and bond of the halogen bond P\u00b7\u00b7\u00b7I\u2014F triad. However, weaker nI-\u03c3PR, nI-\u03c3PH clashes of the alternative pnicogen bond triads also contribute to sub-vdW steric opposition, thereby further offsetting the apparently \u201cexcessive\u201d NBO estimate of donor\u2013acceptor attraction needed to achieve the final equilibrium geometry and net binding energy.Though the donor\u2013acceptor \u0394 energy\u201d between nP, nI, and \u03c3IF NBOs (not shown) similarly serve to rationalize the trends in repulsive donor\u2013donor interactions n(P)\u2192\u03c3*(IF) evaluations of n\u2192\u03c3* donor\u2013acceptor strength in the equilibrium species (E($DEL)n(P)\u2192\u03c3*(IF) potential energy surface in which n\u2192\u03c3* delocalization is absent, as though nature (in accordance with common textbook presentations) failed to include such interactions.Still another widget from the NBO toolbox can be used to exhibit the profound effects of species , one can2\u00b7\u00b7\u00b7IF dissociation, with the nP\u2192\u03c3*IF interaction deleted. Starting from the equilibrium values of the actual RPH2\u00b7\u00b7\u00b7IF species, the distance variables RP\u00b7\u00b7\u00b7I, RIF were optimized to find the new binding energy (\u0394Ebind) and shifts that describe the hypothetical \u201cresonance-free\u201d RPH2\u00b7\u00b7\u00b7IF that lacks nP\u2192\u03c3*IF donor\u2013acceptor attraction. From the resulting values, one sees that characteristic structural and energetic signatures of RPH2\u00b7\u00b7\u00b7IF bonding are lost when the nP\u2192\u03c3*IF interaction is \u201cturned off;\u201d the RPH2 and IF monomers retreat (by circa 1.3\u00c5) to beyond-vdW separation, the characteristic RIF elongation from isolated monomer geometry essentially disappears, and the net binding energy drops precipitously (by >80%) to a remnant value that might plausibly be associated with residual \u201cdipole\u2013dipole\u201d forces. Note that the monomer dipole moments or other aspects of electron density distribution are scarcely altered in the $DEL-reoptimized geometry, and the \u201clost\u201d energy of nP\u2192\u03c3*IF stabilization is negligibly small at the reoptimized separation distance. From the results of such $DEL-deletion calculations, one recognizes nP\u2192\u03c3*IF stabilization to be the unique \u201csmoking gun\u201d that is both necessary and sufficient to bring the supramolecular complex to the actual short-range geometry and other signatures of halogen bonding, which is in direct correspondence to the analogous nB\u2192\u03c3*HA role in H-bonding.n\u2192\u03c3* interactions lead to net binding against the essentially repulsive potential curve for hypothetical monomers that interact without benefit of resonance-type stabilization. 3PH2\u00b7\u00b7\u00b7IF dissociation, showing the potential curve for the fully interacting monomers . The \u201cmissing\u201d resonance-type (purely attractive) interactions are shown as the difference curve . The blue curve exhibits a feeble attractive well beyond vdW contact at RP\u00b7\u00b7\u00b7I \u2248 4.0\u00c5 (presumably due to \u201cdipole\u2013dipole\u201d attraction) but rises steeply as a repulsive steric wall in the sub-vdW region. Already in the region of the initial sub-vdW penetration, the red curve (n\u2192\u03c3* resonance attraction) has achieved sufficient stabilization to oppose thermal fluctuations and yield significant sub-vdW binding in the black curve down to RP\u00b7\u00b7\u00b7I \u2248 2.9 equilibrium separation. Inside this equilibrium distance, the intermolecular n-\u03c3 steric repulsion overcomes n-\u03c3* attraction to give the steeply repulsive inner wall of the full potential, resonance-shifted inside circa 2.5\u00c5. Where such nP-\u03c3IF versus nP-\u03c3*IF \u201ccross-over\u201d occurs will evidently depend on the polarization of the dihalogen bond and is therefore expected to shift to a progressively larger RP\u00b7\u00b7\u00b7I separation (and weakened \u0394Ebind) for other members of the dihalogen series, thereby appearing as a secondary \u201celectrostatic\u201d modulating effect on the overall halogen bonding phenomenon.Still another use of $DEL-deletion techniques is to prepare relaxed-scan potential curves that illustrate how intermolecular 2\u00b7\u00b7\u00b7IF species tends to increase steadily with strength of RPH2\u00b7\u00b7\u00b7IF binding or net QCT. This trend reflects the expected dominance of the nP\u2192\u03c3*IF interaction, which is evidently enhanced . The combined effect of repolarization (iIF) and net charge transfer (QCT) is to give a somewhat irregular pattern to individual atomic charges qI, qF, but all such IF charge descriptors are seen to properly \u201cadd up,\u201d as they must according to the strict logic of a natural population analysis.city iIF ), compar2 monomer, the phosphorus charge qP reflects the still more complex effects of sigma-type induction versus intra- and intermolecular lone pair delocalizations. One can see in the table the evident effect of bonding the phosphorus to more electronegative oxygen (qP \u2248 0.73) or nitrogen (qP \u2248 0.45) rather than carbon (qP \u2248 0.3\u20130.4). However, also important in the present context is the intra- versus intermolecular competition for charge donation from the \u201cbusy\u201d phosphorus lone pair nP to various pi-type acceptor orbitals of the R-substituent versus the intermolecular nP\u2192\u03c3*IF interaction of principal interest.In the RPHbAB}, which balance the nuances of multiple resonance structure contributions to give a single composite measure of A\u00b7\u00b7\u00b7B \u201cconnectivity.\u201d As described elsewhere [NBO 7.0 program [Perhaps the most useful and general descriptors of supramolecular bonding are NRT bond orders {lsewhere ,41,42,43lsewhere versus the \u201cnoncovalent interaction\u201d (dots). Most surprising in these species are the non-vanishing bond orders for the \u201clong-bond\u201d [2\u00b7\u00b7\u00b7IF species, certain P^I long bonds achieve bond orders that rival or exceed those of familiar bP\u00b7\u00b7\u00b7F or bIF linkages .The first three columns of ng-bond\u201d ,77,78 inThe final two columns of E(2)n\u03c3*, \u0394E($DEL)n\u03c3*, Qn\u03c3*,...) intrinsically focus on orbitals of a specific n\u2192\u03c3* \u201cdelocalization\u201d from a specific Lewis structural bonding pattern. Such orbital-specific descriptors are valuable when a single \u201cparent\u201d Lewis structure and \u201cchild\u201d n\u2192\u03c3* delocalization clearly dominate quantum mechanical descriptions of the structural, energetic, and spectroscopic properties of the chosen system. However, for many chemical systems of interest (including the RPH2\u00b7\u00b7\u00b7FI species considered here), multiple n\u2192\u03c3* interactions come into play, corresponding to contributions of alternative resonance\u2013structural bonding patterns that may no longer be \u201cchild-like\u201d compared to a reference parent pattern. Such cases demand the more nuanced NRT descriptors such as interatomic bond orders {bAB} that balance the many possible orbital interactions contributing to the overall resonance hybrid.The principal NBO descriptors of donor\u2013acceptor interactions makes them better adapted to describe empirical correlations with measurable properties. Well-known examples include bond order\u2013bond length (bAB-RAB) [n\u2192\u03c3* limit as well as the more general correlations of NRT bond orders with experimentally measurable properties, spanning the full set of RPH2\u00b7\u00b7\u00b7IF/FI species considered in this work.As could be anticipated from the empirical origins of bond order descriptors in the pre-quantum mechanical conceptions of \u201cmesomerism\u201d theory , the \u201cavbAB-RAB) , bond orbAB-RAB) , and bonbAB-RAB) relation2\u00b7\u00b7\u00b7IF series, in which the nP\u2192\u03c3*IF interaction plays a clearly dominant role, we first consider the mutual correlations of \u0394E(2)n\u03c3*, \u0394E($DEL)n\u03c3*, Qn\u03c3*, bP\u00b7\u00b7\u00b7I descriptors for this interaction. Taking perturbative \u0394E(2)n\u03c3* as the base descriptor for the group, we display (a) \u0394E(2)n\u03c3*-\u0394E($DEL)n\u03c3*, (b) \u0394E(2)n\u03c3*-Qn\u03c3*, and (c) \u0394E(2)n\u03c3*-bP\u00b7\u00b7\u00b7I correlation diagrams in successive panels (a)\u2013(c) of 2 correlation coefficients demonstrate the high quality of these mutual correlations: In the range 0.94\u20130.99 for NBO-specific descriptors, but slightly lower (0.94) for correlation with the NRT bond order bP\u00b7\u00b7\u00b7I (presumably due to the proper inclusion of secondary donor\u2013acceptor interactions with R-substituents that are present only in the latter). The results show that descriptors of this group could be chosen rather interchangeably for correlations with measurable experimental properties when a single donor\u2013acceptor interaction such as nP\u2192\u03c3*IF is clearly dominant.Starting with the RPHRP\u00b7\u00b7\u00b7I or RP\u00b7\u00b7\u00b7F, (b) intramolecular RIF, (c) binding energy \u0394Ebind, or (d) infrared stretching frequency \u03bdIF, we employed the NRT bond order bAB as base descriptor in the correlation diagrams of 2\u00b7\u00b7\u00b7IF and RPH2\u00b7\u00b7\u00b7FI (red: Multi-resonance) series. As examples of bond order\u00adbond length correlations, bP\u00b7\u00b7\u00b7X-RP\u00b7\u00b7\u00b7X correlations for each series, with reasonably high correlations (|\u03c7|2 = 0.92) in each case. bIF-RIF correlation for the dihalogen monomer, with still higher correlation coefficients . As examples of bond order\u2013bond energy correlation, bP\u00b7\u00b7\u00b7X-\u0394Ebind correlations , with similar correlation coefficients (|\u03c7|2 = 0.94) in each series. Finally, as bond order\u2013bond frequency examples, bIF-\u0394\u03bdIF correlations, again with high correlation coefficients . All these correlations suggest the high predictive utility of NRT bond orders for both the tuning effects of R-substituents and the polarity variations of the various dihalogen monomers that govern the broad range of binding energies in these supramolecular species.For more general correlations with measurable experimental properties such as (a) intermolecular 2\u00b7\u00b7\u00b7IF/FI, R = CH3, OH, CF3, CN, NO2), focusing on the electronic origin of what might be identified as a \u201chalogen bond,\u201d \u201cpnicogen bond,\u201d or some combination of both. For this purpose, we employed a broad variety of natural bond orbital (NBO) and natural resonance theory (NRT) descriptors, searching for relationships to hydrogen bonding or other known forms of intermolecular attraction .We have computationally investigated the nature of supramolecular bonding in a series of R-substituted phosphine\u00b7\u00b7\u00b7dihalogen complexes is the dominant feature of halogen-type or pnicogen-type bonding motifs, just as for the dominant nB\u2192\u03c3*HA interaction of B:\u00b7\u00b7\u00b7H\u2014A hydrogen bonding. We obtained evidence both for resonance-type \u201cbond conservation\u201d rules and the complete set of mutually consistent correlations of NBO/NRT descriptors with experimentally measurable properties of RPH2\u00b7\u00b7\u00b7IF/FI complexes that closely parallel those previously demonstrated for H-bonded complexes. We also demonstrate that the removal of n\u2192\u03c3* interactions obliterates the signature features of halogen/pnicogen bonding, whereas such features persist even if envisioned \u201cdipole\u2013dipole\u201d contributions are reversed . Thus, our results establish that resonance-type n\u2192\u03c3* stabilization is both necessary and sufficient for the characteristic structural, energetic, and spectroscopic features of halogen or pnicogen bonding, as was previously demonstrated for H-bonding [Our results show that resonance-type \u201c-bonding ,23. The -bonding ,83. Conn-bonding .n\u2192\u03c3* \u201cfractional bonding\u201d will be found for chalcogen bonds, tetrel bonds, and all other such supramolecular bonding phenomena. Our conclusions thereby extend \u201cresonance covalency\u201d concepts to the entire supramolecular domain of sub-integer bond orders, challenging the common \u201celectrostatic\u201d assumptions that underlie current empirical force field modeling and textbook expositions of supramolecular chemistry. Based on these results, we anticipate that similar unique associations with resonance-type"} +{"text": "The title compound crystallizes with two independent mol\u00adecules in the asymmetric unit. They differ essentially in the orientation of the 4-meth\u00adoxy\u00adphenyl ring with respect to the pyridine ring of the quinoline moiety. 34H28ClN3O3S, contains two independent mol\u00adecules (A and B). They differ essentially in the orientation of the 4-meth\u00adoxy\u00adphenyl ring with respect to the pyridine ring of the quinoline moiety; this dihedral angle is 37.01\u2005(18)\u00b0 in mol\u00adecule A but only 7.06\u2005(17)\u00b0 in mol\u00adecule B. In both mol\u00adecules, the cyclo\u00adhexa\u00adnone ring of the iso\u00adquinoline unit has a half-chair conformation. In the pyrrolo\u00adthia\u00adzole ring system, the pyrrolo ring in mol\u00adecule A has a twisted conformation on the N\u2014C fused bond and an envelope conformation in mol\u00adecule B with the N atom as the flap. The thia\u00adzole rings of both mol\u00adecules have twisted conformations on the N\u2014C fused bond. In the crystal, the A mol\u00adecules are linked by pairs of N\u2014H\u22efO hydrogen bonds, forming inversion dimers with an R22(8) ring motif. These dimers are linked to the B mol\u00adecules by an N\u2014H\u22efN hydrogen bond and a series of C\u2014H\u22efO hydrogen bonds, forming layers lying parallel to the (101) plane. The layers are linked by C\u2014H\u22ef\u03c0 inter\u00adactions and offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.427\u2005(1)\u2005\u00c5], forming a supra\u00admolecular framework. The contribution to the scattering from a region of highly disordered solvent mol\u00adecules was removed with the SQUEEZE routine in PLATON [Spek (2015The asymmetric unit of the title compound, CSpek 2015. Acta Cr In mol\u00adecule A, the dihedral angle between the two rings is 37.01\u2005(18)\u00b0 compared to 7.06\u2005(17)\u00b0 in B. There is also a slight difference in the orientation of the 4-chloro\u00adphenyl ring with respect to the mean plane of the pyrrolo ring, viz. in mol\u00adecule A benzene ring C11A\u2013C16A is inclined to the mean plane of the pyrrol ring (N1A/C1A\u2013C4A) by 86.12\u2005(17)\u00b0, while in mol\u00adecule B the corresponding dihedral angle is 76.92\u2005(17)\u00b0.The mol\u00adecular structure of the two independent mol\u00adecules, A/C21A/C22A/C26A\u2013C28A) in molecule A and (C2B/C21B/C22B/C26B\u2013C28B) in B] of the iso\u00adquinoline unit adopts a half-chair conformation in mol\u00adecule A and a distorted half-chair conformation in mol\u00adecule B , the benzene and pyrrolidine rings make dihedral angles of 3.65\u2005(3) and 3.67\u2005(3)\u00b0 in mol\u00adecules A and B, respectively, while the keto atoms, O2A in mol\u00adecule A and O2B in mol\u00adecule B, deviate from the attached pyrrolidine rings by 0.1116\u2005(1) and 0.0176\u2005(1)\u2005\u00c5, respectively.In the indolin-2-one ring systems -1,2\u2032\u2032(1\u2032\u2032H)-dione methanol solvate -dione chloro\u00adform solvate -6-(4-chloro\u00adbenzyl\u00adidene)-2-(4-meth\u00adoxy\u00adphen\u00adyl)-7,8-di\u00adhydro\u00adquino\u00adlin-5(6H)-one (1\u2005mmol) was added and the reaction mixture was allowed to reflux for a further 14\u2005h. After completion of the reaction, as evident from TLC, the solvent was removed under reduced pressure and the residue washed with ice-cold water (50\u2005ml). The crude product was purified by column chromatography using 90:10 (v/v) petroleum ether\u2013ethyl acetate mixtures to obtain the pure product. The product was dissolved in ethyl acetate and poured into a beaker, covered with perforated film and kept undisturbed. The solvent was allowed to evaporate slowly, yielding colourless block-like crystals after a period of seven days .A mixture of isatin (1.1\u2005mmol) and thia\u00adzolidine-4-carb\u00adoxy\u00adlic acid (1.1\u2005mmol) was taken in 10\u2005ml of aceto\u00adnitrile in a 50\u2005ml round bottom flask and heated to reflux for 2\u2005h. Then (Uiso = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. The residual electron density was difficult to model and therefore, the SQUEEZE routine in PLATON contains approximately 100 electrons global, I. DOI: 10.1107/S2056989019000112/su5469Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019000112/su5469sup3.pdfCSD search. DOI: 1888600CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-chromen-4-yl)\u00adoxy]acetamide derivatives are described and the inter\u00admolecular contacts in the crystals analysed using Hirshfeld surface analysis and two-dimensional fingerprint plots.The crystal structures of two 2-[(2-oxo-2 H-chromen-4-yl)\u00adoxy]acet\u00adyl}piperazin-1-yl)acetamide, C17H19N3O5, (I), and N--2-[(2-oxo-2H-chromen-4-yl)\u00adoxy]acetamide, C20H19NO6, (II), are new coumarin derivatives. In compound (I), the six-membered piperazine adopts a chair conformation. The dihedral angles between the mean planes of the chromene ring and amide plane is 82.65\u2005(7)\u00b0 in (I) and 26.2\u2005(4)\u00b0 in (II). The dihedral angles between the mean planes of the chromene ring and the four planar C atoms of the piperazine ring in (I) and the benzene ring in (II) are 87.66\u2005(6) and 65.0\u2005(4)\u00b0, respectively. There are short intra\u00admolecular contacts in both mol\u00adecules forming S(5) ring motifs, viz. N\u2014H\u22efN and C\u2014H\u22efO in (I), and N\u2014H\u22efO and C\u2014H\u22efN in (II). In the crystals of both compounds, mol\u00adecules are linked by N\u2014H\u22efO hydrogen bonds, forming chains along present within the layers. In the crystal of (II), there are only weak offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.981\u2005(6)\u2005\u00c5] present within the layers. The inter\u00admolecular contacts in the crystals of both compounds have been analysed using Hirshfeld surface analysis and two-dimensional fingerprint plots.The title compounds, 2-(4-{2-[(2-oxo-2 The synthesis, and pharmacological and other properties of coumarin derivatives have been studied and reviewed (Kumar Q = 0.561\u2005(2)\u2005\u00c5, \u03b8 = 0.67\u2005(2)\u00b0 and \u03c6 = 149\u2005(2)\u00b0], and is positioned anti with respect to the C\u2014N rotamer of the amide. Nevertheless, because the asymmetry of the chromene residue, the anti conformation can assume a cis or trans geometry with respect to the relative position of the carbonyl O atom of the carboxamide and the C10\u2014C11 and C16\u2014C17 bonds. Both compounds exhibit a cis relation between these bonds, as can be seen in Figs. 1S(5) ring motifs. In addition, the carbonyl oxygen atom O4 acts as the acceptor for a weak inter\u00adaction with a hydrogen bond of the exocyclic piperazine ring, forming a second S(5) ring motif in (I)S(5) ring motif in (II)The mol\u00adecular structures of compounds (I)The values of the dihedral angles between the mean planes of the planar chromene ring system and the amide plane (C10/C11/O4/N1) are 82.65\u2005(7) and 26.2\u2005(4)\u00b0 in compounds (I)N-(substituted phen\u00adyl)-4H-chromene-2-carboxamides and C(7) chains formed via the C10\u2014H10A\u22efO5iii and N3\u2014H3A\u22efO4i hydrogen bonds, respectively. A C\u2014H\u22ef\u03c0 inter\u00adaction is also present within the layer (Table\u00a01Cg2\u22efCg2iv = 3.691\u2005(1)\u2005\u00c5, inter\u00adplanar distance = 3.490\u2005(1)\u2005\u00c5, offset = 1.20\u2005\u00c5; Cg2 is the centroid of the O1/C1\u2013C9 ring; symmetry code: (iv) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01].In the crystal of (I)ne Fig.\u00a03. The C14r Table\u00a01. An offsi hydrogen bonds, forming chains along the [010] direction, see Fig.\u00a04ii, C5\u2014H5\u22efO4i and C15\u2014H15\u22efO4iii hydrogen bonds, forming layers parallel to the ab plane \u2005\u00c5, inter\u00adplanar distances = 3.638\u2005(4) and 3.508\u2005(4)\u2005\u00c5, offset 0 1.88\u2005\u00c5; Cg2 and Cg3 are the centroids of rings C1\u2013C6 and C13\u2013C18, respectively; symmetry code: (iv) \u2212x\u00a0+\u00a01, y\u00a0+\u00a0z\u00a0+\u00a01].In the crystal of (II)ne Fig.\u00a04. Within et al., 2019et al., 2007CrystalExplorer17 through white to blue . The dnorm surface was mapped over a fixed colour scale of \u22120.544 (red) to 1.418 (blue) for compound (I)The Hirshfeld surfaces of compounds (I)b) and O\u22efH/H\u22efO at 35.3% , followed by the C\u22efH/H\u22efC contacts at 11.8% . For compound (II)b) and O\u22efH/H\u22efO at 32.4% , followed by the C\u22efH/H\u22efC contacts at 16.7% . In both compounds, the H\u22efH inter\u00admolecular contacts predominate, followed by the O\u22efH/H\u22efO contacts. However the C\u22efH/H\u22efC contacts are significantly different 11.8% cf. 16.7% for (I)The fingerprint plots are given in Figs. 83% Fig.\u00a08c, follo8% Fig.\u00a08d. For c4% Fig.\u00a09c, follo7% Fig.\u00a09d. In boet al., 2016H-chromen-4-yl)\u00adoxy]acetamide derivatives gave two hits. They include 2-[(2-oxo-2H-chromen-4-yl)\u00adoxy]-N-(1-phenyl\u00adeth\u00adyl)acetamide -2-[(2-oxo-2H-chromen-4-yl)\u00adoxy]prop\u00adan\u00ada\u00admide structures gave 35 hits. They include four reports, CSD refcodes AMYROL [Kato, 1970et al., 1982et al., 1985et al., 2011et al., 2014et al., 2017meta-substituted coumarin esters cyclo\u00adpropyl sulfamate (Morin Compound (I) To a solution of 1 equiv. of 4-(2-(piperazine-1-yl)eth\u00adoxy)-2H-chromen-2-one (1.0\u2005g) in di\u00adchloro\u00admethane (10\u2005ml) at 273\u2013278\u2005K were added tri\u00adethyl\u00adamine followed by iodo\u00adacetamide , and the reaction mixture was stirred at the same temperature for 1\u2005h. On completion of the reaction (monitored by TLC), the reaction mixture was diluted with di\u00adchloro\u00admethane and water (10\u2005ml). The organic layer was separated and washed with brine solution. It was then dried over anhydrous sodium sulfate, filtered and then evaporated under reduced pressure giving compound (I)Compound (II)N,N-Diiso\u00adpropyl\u00adethyl\u00adamine was added to a mixture of 2-(2-oxo-2H-chromen-4-yl\u00adoxy)acetic acid , 1-ethyl-3-(3-di\u00admethyl\u00adamino\u00adprop\u00adyl)carbodi\u00adimide , 1-hy\u00addroxy\u00adbenzotriazole hydrate , 2,4-di\u00admeth\u00adoxy\u00adbenzyl\u00adamine in N,N-di\u00admethyl\u00adformamide (5\u2005ml) at 273\u2013278\u2005K. The temperature of the mixture was raised to ambient temperature and stirred for 8\u2005h. Progress of the reaction was monitored by TLC (mobile phase: ethyl acetate/hexa\u00adne). After completion of the reaction, the mixture was poured into ice\u2013water and compound (II)Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019003736/su5482sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989019003736/su5482Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019003736/su5482IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989019003736/su5482Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019003736/su5482IIsup5.cmlSupporting information file. DOI: 1891497, 1891496CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structures of three salicyaldoxime compounds are discussed together with Hirshfeld surface and fingerprint analyses. 8H9NO2, 1, 2,4-di\u00adhydroxy\u00adbenzaldehyde oxime, C7H7NO3, 2, and 2-hy\u00addroxy-4-meth\u00adoxy\u00adbenzaldehyde oxime, C8H9NO3, 3, are discussed. In each compound, the hydroxyl groups are essentially coplanar with their attached phenyl group. The inter\u00adplanar angles between the C=N\u2014O moieties of the oxime unit and their attached phenyl rings are 0.08\u2005(9), 1.08\u2005(15) and 6.65\u2005(15)\u00b0 in 1, 2 and 3, respectively. In all three mol\u00adecules, the 2-hy\u00addroxy group forms an intra\u00admolecular O\u2014H\u22efN(oxime) hydrogen bond. In compound (1), inter\u00admolecular O\u2014H(oxime)\u22efO(hydrox\u00adyl) hydrogen bonds generate R22(14) dimers, related by inversion centres. In compound 2, inter\u00admolecular O\u2014H(oxime)\u22efO(4-hy\u00addroxy) hydrogen bonds generate C9 chains along the b-axis direction, while O\u2014H(4-hydrox\u00adyl)\u22efO(2-hydrox\u00adyl) inter\u00adactions form zigzag C6 spiral chains along the c-axis direction, generated by a screw axis at 1, y, 1/4: the combination of the two chains provides a bimolecular sheet running parallel to the b axis, which lies between 0\u20131/2 c and 1/2\u20131 c. In compound 3, similar C9 chains, along the b-axis direction are generated by O\u2014H(oxime)\u22efO(4-meth\u00adoxy) hydrogen bonds. Further weaker, C\u2014H\u22ef\u03c0 (in 1), \u03c0\u2013\u03c0 (in 2) and both C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions (in 3) further cement the three-dimensional structures. Hirshfeld surface and fingerprint analyses are discussed.The crystal structures of three salicyaldoxime compounds, namely 2-hy\u00addroxy-4-methyl\u00adbenzaldehyde oxime, C RCH=NOH, are found in many biologically active compounds with different substituents in the 4-position, namely a methyl group, a hy\u00addroxy group and a meth\u00adoxy group, respectively, in compounds, 1, 2 and 3. A frequent finding for salicylaldoxime derivatives is the formation of inversion-related et al., 2016The compounds described herein are all salicylaldoxime derivatives geometry. Bond angles and bond lengths in the phenyl and oxime fragments are all in the expected ranges.There are no unusual features in the mol\u00adecular structures. Compound 1 Fig.\u00a01 crystallt Figs. 2 and 3 \u25b8,1, the hydroxyl group is essentially coplanar with its attached phenyl group [displaced by 0.020\u2005(1)\u2005\u00c5], while the inter\u00adplanar angle between the C=NO moiety of the oxime unit and the attached phenyl rings is 0.08\u2005(9)\u00b0. In compound 2, the hydroxyl groups lie essentially within the phenyl ring plane [O atoms deviate by \u22120.003\u2005(1) and 0.006\u2005(1)\u2005\u00c5], while the inter\u00adplanar angle between the C=NO moiety of the oxime unit and the attached phenyl rings is 1.08\u2005(15)\u00b0. In compound 3, the inter\u00adplanar angle between the C=NO moiety of the oxime unit and the attached phenyl rings is 6.65\u2005(15)\u00b0.In compound In all three mol\u00adecules, an intra\u00admolecular O2\u2014H2\u22efN12 hydrogen bond Tables 1\u20133 \u25b8 \u25b8 f1, mol\u00adecules are linked by O13\u2014H13 \u22efO2 hydrogen bonds into inversion-related via the intra\u00admolecular hydrogen bond) dimers are linked into two-mol\u00adecule-wide chains, propagating in the a-axis direction by pairs of O13\u2014H13\u22efO13 hydrogen bonds, thereby creating ii hydrogen bond are rather long [2.611\u2005(16)\u2005\u00c5] with a small angle of 100.8\u2005(12)\u00b0. However, such data fits well with published findings for H2O2 rings: a recent CSD \u22efO4(4-hy\u00addroxy)ii hydrogen bonds, propagating in the direction of the b axis, see Fig.\u00a05C6 spiral chain formed from O4\u2014H4\u22efO2i hydrogen bonds, see Fig.\u00a06b axis which lies between 0\u2013\u00bd c and \u00bd\u20131 c. These sheets are further linked by moderately strong \u03c0\u2013\u03c0 stacking inter\u00adactions, involving all the phenyl rings in the sheet: the Cg\u22efCg separation is 3.7242\u2005(13)\u2005\u00c5 with a phenyl ring slippage of 1.586\u2005\u00c5. The lack of an 2 is apparent and results from the preferential inter\u00adaction of the oxime group with the 4-hydroxyl group rather than with the 2-hy\u00addroxy group.Compound 3, C9 chains are generated from O13\u2014H13\u22efO41(meth\u00adoxy)i hydrogen bonds, which propagate in the direction of the b axis, see Fig.\u00a072, but involving the meth\u00adoxy oxygen atom O41 involved instead of the hy\u00addroxy oxygen O4. Inter\u00adestingly, the parameters of the two hydrogen bonds in the chains of compound 2 and 3 are very similar. The chains in compound 3 are linked into a two-dimensional array by C11\u2014H11\u22efCg . The former hydrogen bond forms C9 chains, forming a corrugated ribbon which runs parallel to the a axis.In compound g Table\u00a03 and \u03c0\u2013\u03c0 CrystalExplorer3.1 of the formation of intra\u00admolecular O\u2014H\u22efNO(oxime) hydrogen bonds involv\u00ading the ortho hydroxyl group. In addition, this hydroxyl group is also most frequently involved in inter\u00admolecular inter\u00adactions producing inversion-related et al., 2003et al., 2006bR = 2-HO-5-MeOC6H3, producing a C5 chain from O\u2014H(oxime)\u22efO(2-hydrox\u00adyl) hydrogen bonds; Pfluger et al., 1978R = 2-HO-5-tBu-C6H3, producing a C5 chain from O\u2014H(oxime)\u22efO(2-hydrox\u00adyl) hydrogen bonds; White et al., 2015aR = 2-HO-3-Me-5-(piperin-1-yl-CH2)-C6H2, producing a C9 chain from O\u2014H(oxime)\u22efN(piperin\u00adyl) hydrogen bonds; White et al., 2015bR = 2-HO-3-(piperin-1-ylmeth\u00adyl)-5-tBu-C6H2, producing a C9 chain from O\u2014H(oxime)\u22efN(piperin\u00adyl) hydrogen bonds; Forgan et al., 2010A survey of the Cambridge Structural Database and 2-HO-5-MeOC6H3CH=N\u2014OH = 1.2\u20131.5Uiso(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989018013361/qm2128sup1.cifCrystal structure: contains datablock(s) 1, 2, 3, global. DOI: 10.1107/S2056989018013361/qm21281sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989018013361/qm21282sup3.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S2056989018013361/qm21283sup4.hklStructure factors: contains datablock(s) 3. DOI: Click here for additional data file.10.1107/S2056989018013361/qm21281sup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018013361/qm21282sup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018013361/qm21283sup7.cmlSupporting information file. DOI: 1868656, 1868655, 1868654CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The correct citation is: M\u00e6kel\u00e6 MJ, Pfuhl G (2019) Deliberate reasoning is not affected by language. PLoS ONE 14(1): e0211428."} +{"text": "ER) stress shifts the unfolded protein response signaling from cell survival to cell death, although the switching mechanism remains unclear. Here, we report that mitochondrial ubiquitin ligase (MITOL/MARCH5) inhibits ER stress\u2010induced apoptosis through ubiquitylation of IRE1\u03b1 at the mitochondria\u2010associated ER membrane (MAM). MITOL promotes K63\u2010linked chain ubiquitination of IRE1\u03b1 at lysine 481 (K481), thereby preventing hyper\u2010oligomerization of IRE1\u03b1 and regulated IRE1\u03b1\u2010dependent decay (RIDD). Therefore, under ER stress, MITOL depletion or the IRE1\u03b1 mutant (K481R) allows for IRE1\u03b1 hyper\u2010oligomerization and enhances RIDD activity, resulting in apoptosis. Similarly, in the spinal cord of MITOL\u2010deficient mice, ER stress enhances RIDD activity and subsequent apoptosis. Notably, unresolved ER stress attenuates IRE1\u03b1 ubiquitylation, suggesting that this directs the apoptotic switch of IRE1\u03b1 signaling. Our findings suggest that mitochondria regulate cell fate under ER stress through IRE1\u03b1 ubiquitylation by MITOL at the MAM.Unresolved endoplasmic reticulum ( Various physiological and pathological changes require enhanced ER functions, such as protein folding, removal of unfolded proteins, and reduction of oxidative stress. An imbalance in ER homeostasis causes ER stress and triggers the unfolded protein response (UPR), which is initiated by three ER proteins, PERK, ATF6, and IRE1\u03b1. The UPR is involved in the recovery of ER homeostasis through ERAD and ER chaperones and plays a central role in the lipid metabolism and Caet\u00a0al, et\u00a0al, et\u00a0al, et\u00a0al, We have previously identified mitochondrial ubiquitin ligase (MITOL/MARCH5) and demonstrated that MITOL ubiquitylates mitofusin 2 (Mfn2) and enhances its GTPase activity, resulting in the tethering between the ER and mitochondria rather than mitochondrial fusion 481 of IRE1\u03b1 by adding a K63\u2010linked polyubiquitin chain, which in turn suppresses the activity and oligomerization of IRE1\u03b1. Here, we provide evidence that MITOL can directly regulate IRE1\u03b1 through the MAM and thus play an important role in determining cell fate under ER stress.mitol/march5 gene \u2010inducible MITOL\u2010knockout MEFs by flanking two loxP sequences with exon 2 of the ells Fig\u00a0E\u2013G, sugg2+ in the ER was increased in MITOL\u2010KO MEFs showed abnormal morphology of the ER but not of the mitochondria Fig\u00a0H and I. MEFs Fig\u00a0J. We firMEFs Fig\u00a0A. MITOL\u2010MEFs Fig\u00a0B. To furMEFs Fig\u00a0C. In conMEFs Fig\u00a0C. These et\u00a0al, ER stress has been shown to trigger apoptosis via mitochondrial permeability transition pore was used to select the genomic sequence target in human MITOL (5\u02b9\u2010CCAAGCCCTACAGCAGATGC\u20103\u02b9). Oligo pairs encoding 20\u2010nt guide sequences were annealed and ligated into the plasmid pX330 (purchased from Addgene). HEK293 cells were transfected and MITOL\u2010KO clones selected by serial dilution.MEFs, HEK293, and COS\u20107 cells and HeLa cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin. Cells were transfected with either Lipofectamine 3000 (Invitrogen) or RNAiMax (Invitrogen) according to the manufacturer's protocol. The following siRNAs were used: siIRE1\u03b1 #1: sense, 5\u02b9\u2010CGGGCUCCAUCAAGUGGACUUUAAATT\u20103\u02b9, antisense, 5\u02b9\u2010UUUAAAGUCCACUUGAUGGAGCCCGTT\u20103\u02b9; siIRE1\u03b1 #2: sense, 5\u02b9\u2010AAGAUGGACUGGCGGGAGATT\u20103\u02b9, antisense, 5\u02b9\u2010UCUCCCGCCAGUCCAUCUUTT\u20103\u02b9; siJNK1 #1: sense, 5\u02b9\u2010AAAGAAUGUCCUACCUUCUTT\u20103\u02b9, antisense, 5\u02b9\u2010AGAAGGUAGGACAUUCUUUTT\u20103\u02b9; and siJNK1 #2: sense, 5\u02b9\u2010GCAGAAGCAAACGUGACAACATT\u20103\u02b9, antisense, 5\u02b9\u2010UGUUGUCACGUUUGCUUCUGCTT\u20103\u02b9. siPACS2 and siMfn2 were described previously was subjected to further separation by 30% Percoll gradient centrifugation at 95,000\u00a0\u00d7\u00a0g for 30\u00a0min at 4\u00b0C. A low\u2010density band (denoted as the MAM fraction) was purified. The pure mitochondrial fraction in the high\u2010density band was resuspended in HB and pelleted by centrifugation . Microsomes were pelleted from the mitochondrial supernatant by centrifugation at 100,000\u00a0\u00d7\u00a0g for 1\u00a0h. Cytosol was recovered as the supernatant from this centrifugation.Isolation of crude mitochondria or pure MAM was performed as described previously and subjected to reverse transcription to cDNA using ReverTra Ace qPCR RT Kit (TOYOBO), following the manufacturer's protocol. PCR was performed using a THUNDERBIRD SYBR qPCR Mix (TOYOBO). The PCR conditions were as follows: 95\u00b0C for 1\u00a0min followed by 40 cycles at 95\u00b0C for 15\u00a0s, 60\u00b0C for 30\u00a0s, and 72\u00b0C for 60\u00a0s. RT\u2013PCR was performed using miScript SYBR Green PCR Kit and miScript Primer Assays (Qiagen).atf4: forward, 5\u02b9\u2010GGA CAGATTGGATGTTGGAGAAAATG\u20103\u02b9, reverse, 5\u02b9\u2010GGAGATGGCCAATTGGGTTCAC\u20103\u02b9; caspase\u20102: forward, 5\u02b9\u2010 CCACAGATGCTACGGAACA\u20103\u02b9, reverse, 5\u02b9\u2010 GCTGGTAGTGTGCCTGGTAA\u20103\u02b9; chop: forward, 5\u02b9\u2010CATACACCACCACACCTGAAAG\u20103\u02b9, reverse, 5\u02b9\u2010CCGTTTCCTAGTTCTTCCTTGC\u20103\u02b9; dr5: forward, 5\u02b9\u2010CTGTGCTACAGGCTGTCTTTG\u20103\u02b9, reverse, 5\u02b9\u2010GTACTGGCCTGCTAGACAG\u20103\u02b9; xbp1s: forward, 5\u02b9\u2010GTGTCAGAGTCCATGGGA\u20103\u02b9, reverse, 5\u02b9\u2010GAGTCCGCAGCAGGTG\u20103\u02b9; edem: forward, 5\u02b9\u2010AAGCCCTCTGGAACTTGCG\u20103\u02b9, reverse, 5\u02b9\u2010AACCCAATGGCCTGTCTGG\u20103\u02b9; Sec61: forward, 5\u02b9\u2010CTATTTCCAGGGCTTCCGAGT\u20103\u02b9, reverse, 5\u02b9\u2010AGGTGTTGTACTGGCCTCGGT\u20103\u02b9; herp: forward, 5\u02b9\u2010CATGTACCTGCACCACGTCG\u20103\u02b9, reverse, 5\u02b9\u2010GAGGACCACCATCATCCGG\u20103\u02b9; col6a1: forward, 5\u02b9\u2010TGCTCAACATGAAGCAGACC\u20103\u02b9, reverse, 5\u02b9\u2010TTGAGGGAGAAAGCTCTGGA\u20103\u02b9; blos1: forward, 5\u02b9\u2010CAAGGAGCTGCAGGAGAAGA\u20103\u02b9, reverse, 5\u02b9\u2010GCCTGGTTGAAGTTCTCCAC\u20103\u02b9; hgsnat: forward, 5\u02b9\u2010TCTCCGCTTTCTCCATTTTG\u20103\u02b9, reverse, 5\u02b9\u2010CGCATACACGTGGAAAGTCA\u20103\u02b9; pdgfrp: forward, 5\u02b9\u2010AACCCCCTTACAGCTGTCCT\u20103\u02b9, reverse, 5\u02b9\u2010TAATCCCGTCAGCATCTTCC\u20103\u02b9; txnip: forward, 5\u02b9\u2010TCAAGGGCCCCTGGGAACATC\u20103\u02b9, reverse, 5\u02b9\u2010GACACTGGTGCCATTAAGTCAG\u20103\u02b9; and gapdh: forward, 5\u02b9\u2010AACTTTGGCATTGTGGAAGG\u20103\u02b9, reverse, 5\u02b9\u2010GGATGCAGGGATGATGTTCT\u20103\u02b9. Primers for xbp1s were designed to amplify the region containing sequences removed by IRE1\u03b1.The following primers were used: g, the precipitate and supernatant were analyzed by immunoblotting.To extract cholesterol\u2010enriched lipid raft from MAM, 100\u00a0\u03bcg of crude mitochondria was incubated for 10\u00a0min with indicated final concentrations of digitonin on ice. After centrifugation at 8,000\u00a0\u00d7\u00a0g for 20\u00a0min at 4\u00b0C. The supernatant was subjected to immunoprecipitation using indicated antibodies. To evaluate the ubiquitylation level of proteins, cells were solubilized with RIPA lysis buffer and centrifuged at 20,000\u00a0g for 20\u00a0min at 4\u00b0C. The supernatant was sonicated 10\u00a0s and subjected to immunoprecipitation using indicated antibodies.To examine the protein interaction, cells were solubilized with NP\u201040 lysis buffer and centrifuged at 20,000\u00a02 following the manufacturer's protocol.Whole lysates were separated by SDS\u2013PAGE and transferred to the PVDF membranes (Millipore). The blots were probed with indicated antibodies, and protein bands on the blot were visualized by the enhanced chemiluminescence reagent (Millipore). Phos\u2010tag\u2010PAGE was performed using 7% SDS\u2013PAGE minigels containing 10\u00a0\u03bcM Phos\u2010tag (Wako) in the presence of 100\u00a0\u03bcM MnCl2, 4\u00a0mM ATP, 200\u00a0ng of E1 , 600\u00a0ng of UbcH5b (Biomol), and 5\u00a0\u03bcg of His\u2010Ub (Biomol) for 2\u00a0h at 30\u00b0C and then terminated with 3\u00a0\u00d7\u00a0SDS sample buffer.IRE1\u03b1\u2010FLAG and MITOL\u2010HA were purified from HEK293 cells by immunoprecipitation. The purified proteins were incubated with reaction buffer containing 50\u00a0mM Tris\u2013HCl, pH 7.4, 2\u00a0mM MgClet\u00a0al, To determine the specific domains required for the interaction between MITOL and IRE1\u03b1, GST\u2010fusion deletion mutants were prepared using glutathione high\u2010capacity magnetic agarose beads (Sigma) as described previously , and analyzed using an Olympus IX81 confocal fluorescence microscope. For classification of the morphology of mitochondrial network, four stacks of 0.2\u00a0\u03bcm were acquired for cells stained with anti\u2010Tom20 antibody using 60\u00d7 objective. Images were compiled and classified in a blinded manner as normal in which the majority of mitochondria were connected in the cell or abnormal in which many mitochondria exhibited the spherical structure or showed the disconnected network overall. For quantification of the morphology of the ER network, four stacks of 0.2\u00a0\u03bcm were acquired for cells transfected with mC\u2010Sec61\u03b2 using 60\u00d7 objective and classified in a blinded manner as normal morphology with reticular\u2010like ER network or abnormal morphology either with a more sheet\u2010like ER network overall or with at least three aggregated parts in the ER network. Percentages of cells with abnormal ER network in each condition were calculated from 50 cells in each independent experiment.In Situ Cell Death Detection Kit (Roche). For Nissl, staining was performed using 0.5% cresyl violet. The sections were observed using an All\u2010in\u2010One Fluorescence Microscope BZ\u20109000.Spinal cord was fixed with 10% formalin neutral buffer solution (Wako), and 5\u2010\u03bcm\u2010thick paraffin sections were prepared. TUNEL staining was performed using 2+ imaging was performed as described previously and re\u2010fed with HBSS. G\u2010CEPIA1er signals of transfected cells were monitored per 500\u2010ms interval during 60\u00a0s. At 20\u00a0s after Ca2+ imaging started, these cells were co\u2010incubated with 5\u00a0\u03bcM ionomycin. The level of Ca2+ in the ER was obtained as \u0394F values, calculated from F0 minus Fmax. F0 values were obtained by the average intensity of G\u2010CEPIA1er in each cell before ionomycin treatment. Fmax was calculated from the average intensity of G\u2010CEPIA1er in each cell treated with ionomycin. The intensities of G\u2010CEPIA1er were measured using ImageJ.ER Caet\u00a0al, g for 20\u00a0min at 4\u00b0C. The supernatant was centrifuged at 250,000\u00a0g for 12\u00a0h at 4\u00b0C through 20\u201340% sucrose gradient , prepared freshly by progressively layering higher to lower density sucrose fractions in 5% increments. Each 4\u00a0ml gradient was divided evenly into 16 fractions (250\u00a0\u03bcl each), and then, aliquots of fractions 2\u201313 were analyzed by immunoblotting.Sucrose gradient assay was performed as described previously in saline.MITOLAnnexin V\u2010FITC staining was performed with Annexin V\u2010FITC apoptosis detection kit (BioVision) according to the manufacturer's protocol. For the detection of mitochondrial depolarization, cells were incubated with 10\u00a0nM tetramethylrhodamine methyl ester (TMRM) for 30\u00a0min at 37\u00b0C and then washed with PBS. The fluorescence of TMRM was analyzed by flow cytometer (Becton Dickinson).Luciferase assay was performed with dual\u2010luciferase reporter assay system (Promega) according to the manufacturer's protocol.Cell viability and cell toxicity were monitored using the Cell Counting Kit\u20108 (Dojindo) and Cytotoxicity LDH Assay Kit\u2010WST (Dojindo), respectively, according to the manufacturer's protocol.t\u2010test. The number of independent experiments is shown as n.All results are expressed as mean\u00a0\u00b1\u00a0SD. Obtained data were compared between independent experiments using either two\u2010tailed Student's Flox/Flox mice and MITOL\u2010related MEFs. TI and TU generated IRE1\u03b1\u2010KO MEFs. TT, TF, and NM contributed to the understanding of MITOLnestin mice phenotype. KT and SY wrote the manuscript with contribution from RI.KT and SY designed and carried out the experiments. SN analyzed the data. AU carried out qRT\u2013PCR and cultured cell analysis with contribution from IS and NI. SI generated MITOLThe authors declare that they have no conflict of interest.Expanded View Figures PDFClick here for additional data file.Review Process FileClick here for additional data file."} +{"text": "The conformation of the title compound is discussed and compared to those of related structures. In the crystal, mol\u00adecules of the title compound are assembled into layers parallel to the 28H22N4O9, exhibits crystallographically imposed twofold rotational symmetry, with a dihedral angle of 66.0\u2005(2)\u00b0 between the planes of the two central benzene rings bounded to the central oxygen atom. The dihedral angle between the planes of the central benzene ring and the terminal phenol ring is 4.9\u2005(2)\u00b0. Each half of the mol\u00adecule exhibits an imine E configuration. An intra\u00admolecular O\u2014H\u22efN hydrogen bond is present. In the crystal, the mol\u00adecules are linked into layers parallel to the ab plane via C\u2014H\u22efO hydrogen bonds. The crystal studied was refined as a two-component pseudomerohedral twin.The mol\u00adecule of the title compound, C The dihedral angle between the benzene and 4-meth\u00adoxy-2-nitro\u00adphenol rings in the same half of the mol\u00adecules is 4.9\u2005(2)\u00b0, indicating an almost coplanar arrangement of the benzene and phenol rings. The sp2-hybridized character of atoms N1 and C7 is confirmed by the N1\u2014C7 [1.287\u2005(6)\u2005\u00c5] bond length and C7\u2014N1\u2014C8 [121.9\u2005(4)\u00b0] and N1\u2014C7\u2014C6 [121.7\u2005(4)\u00b0] bond angles \u00b0. In the mol\u00adecule, atom N1 of the imine moiety acts as a hydrogen-bond acceptor for the adjacent phenol group, forming an intra\u00admolecular O\u2014H\u22efN hydrogen bond with an S(6) ring motif -N,N\u2032-[oxybis]bis\u00ad(1-phenyl\u00admethanimine) moiety with different substituents were found. The reference moiety is illustrated in Fig.\u00a031R) together with the dihedral and torsion angles for oxybisbenzenyl moiety in these structures are tabulated in Table\u00a02viz. non-coplanar [dihedral 2 = 18.0\u201373.5\u00b0] and nearly coplanar [dihedral 2 = 4.8\u20139.9\u00b0]. In all of these structures, the imine C=N double bond adopts an E configuration with torsion angles corresponding to C6\u2014C7\u2014N1\u2014C8 in the range 172.9\u2013180.0\u00b0.In a search of the Cambridge Structure Database C, 60.16; H, 3.93; N, 10; found: C, 59.04; H, 3.85; N, 9.90%. 1H NMR : \u03b4 10.23 , \u03b4 9.12 , \u03b4 7.69\u20137.21 , \u03b4 3.83 . 13C NMR : \u03b4 161.69 (C=N), \u03b4 156.21\u2013114.96 (C-aromatic), \u03b4 56.25 (OCH3). IR (KBr pellets \u03c5max/cm\u22121): 3441 \u03c5(OH), 3109 \u03c5, 2956 \u03c5(CH3), 1598 \u03c5(C=N), 1529 \u03c5, 1497 \u03c5, 1326 \u03c5, 1257 \u03c5, 1194 \u03c5, 1056 \u03c5(C\u2014N), 979 \u03c5.To a sample of 2-hy\u00addroxy-5-meth\u00adoxy-3-nitro\u00adbenzaldehyde dissolved in 25.0\u2005mL of methanol, 0.20\u2005mL of glacial acetic acid were added, and the mixture was refluxed for 30\u2005min. A solution of 4,4\u2032-oxydianiline in 20.0\u2005mL of methanol was added dropwise under stirring to the aldehyde solution. The resulting deep-red solution was refluxed for 4\u2005h with stirring. The reaction scheme is shown in Fig.\u00a04Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C) for methyl H atoms. A rotating model was used for the methyl group. The crystal investigated was refined as a two-component pseudomerohedral twin resulting from a 180\u00b0 rotation about the [001] reciprocal lattice direction, with a twin ratio of 0.977\u2005(3):0.023\u2005(3).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019016852/rz5267sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019016852/rz5267Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019016852/rz5267Isup3.cmlSupporting information file. DOI: 1445336CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure and supra\u00admolecular inter\u00adactions between a hy\u00addroxy-functionalized pillarene and 1-octa\u00adnamine guest mol\u00adecule, which forms an inter\u00adesting host\u2013guest system, are reported. 44H48O10\u00b7C8H19N\u00b7H2O. The guest compound is stabilized inside the cavity by hydrogen-bonding and C\u2014H\u22ef\u03c0 inter\u00adactions. The water mol\u00adecule in the asymmetric unit mediates the formation of a supra\u00admolecular dimer by hydrogen-bonding inter\u00adactions. These functionalized-pillararene hosts expand the possibility of exploring more supra\u00admolecular inter\u00adactions with various guest species.Co-crystallization of a monofunctionalized hy\u00addroxy pillar[5]arene with 1-octa\u00adnamine resulted in the formation of an inclusion complex where the alkyl chain is threaded in the macrocycle cavity, namely 1,2,3,4--5-(1-hy\u00addroxy-4-meth\u00adoxy)-pillar[5]arene\u20131-octa\u00adnamine\u2013water (1/1/1), C Unlike cone-shaped calixarene or resorcinarene-type structures, the pillararenes have a tabular cavity, which makes them inter\u00adesting mol\u00adecular hosts. It is well known that pillar[5]arenes exhibit an outstanding ability to selectively bind different kinds of guest mol\u00adecules and thus are valuable chemical entities in the areas of host\u2013guest systems and mol\u00adecular recognition . Furthermore, the OH\u2013 group in pillararenes could involve hydrogen bonding with guest mol\u00adecules and/or with neighboring pillararenes, which makes them valuable compounds in mol\u00adecular recognition and supra\u00admolecular chemistry. We have recently reported details of the host\u2013guest complexation between mono-hy\u00addroxy-pillar[5]arenes with long-chain alkyl alcohol guests . It was observed that the encapsulation characteristics of the pillar[5]arene was affected by the presence of the hy\u00addroxy group, resulting in the formation of a 1:2 complex with long-chain alkyl alcohols.The pillar[5]rene system having one hy\u00addroxy group is inter\u00adesting because this OH\u2013 function is susceptible for further chemical transformation (Al-Azemi et Pil-OH) and 1-octa\u00adnamine (2OctNH). The structural features and supra\u00admolecular host\u2013guest inter\u00adactions of this co-crystalline system (2Pil-OH\u00b7OctNH) has been addressed and discussed.In this work we report the crystal structure of the inclusion complex consisting of 1-(1-hy\u00addroxy-4-meth\u00adoxy)-2,3,4,5--pillar[5]arene has a rigid three-dimensional macrocyclic architecture with a wide cavity having a penta\u00adgonal shape. The 1-octa\u00adnamine mol\u00adecule is threaded inside the pillararene cavity and one water is included in asymmetric unit, displaying strong hydrogen-bonding inter\u00adactions with the amino group of the guest mol\u00adecule inside the cavity and the hy\u00addroxy group on the pillararene system via O11\u2014H11A\u22efN1 and O11\u2014H11B\u22efO1 bonds respectively \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01] and O11\u2014H11\u22efO1 hydrogen-bonding inter\u00adactions x\u00a0\u2212\u00a01, y, z]. A weak C\u2014H\u22efO type pillararene\u2013pillararene inter\u00adaction is also observed .In the title inclusion complex, the water mol\u00adecule mediates the formation of supra\u00admolecular dimers through O1s Table\u00a01, as illuet al., 2018The synthesis of 1-(1-hy\u00addroxy-4-meth\u00adoxy)-2,3,4,5-pillar[5]arene has been reported previously = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018010034/dx2005sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018010034/dx2005Isup4.hklStructure factors: contains datablock(s) I. DOI: 1855261CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound possesses crystallographically imposed twofold symmetry with the two C atoms of the central benzene ring and the C atom of its methyl substituent lying on the twofold rotation axis. The two dansyl groups are twisted away from the plane of methyl\u00adphenyl bridging unit in opposite directions. The crystal packing features weak C\u2014H\u22efO hydrogen bonds. 31H30N2S2O6, possesses crystallographically imposed twofold symmetry with the two C atoms of the central benzene ring and the C atom of its methyl substituent lying on the twofold rotation axis. The two dansyl groups are twisted away from the plane of methyl\u00adphenyl bridging unit in opposite directions. The three-dimensional arrangement in the crystal is mainly stabilized by weak hydrogen bonds between the sulfonyl oxygen atoms and the hydrogen atoms from the N-methyl groups. Stacking of the dansyl group is not observed. From the DFT calculations, the HOMO\u2013LUMO energy gap was found to be 2.99\u2005eV and indicates n\u2192\u03c0* and \u03c0\u2192\u03c0* transitions within the mol\u00adecule.The title compound, C Such modified oligonucleotides can be used to prepare and detect the sequence of fluoro\u00adgenic probes in DNA rings, Fig.\u00a01O-dansyl groups on either side of a bridging methyl\u00adphenyl ring that is essentially planar. The S1\u2014O1\u2014C4\u2014C3 torsion angle is 72.98\u2005(16)\u00b0 with the methyl\u00adphenyl ring plane. The S1 sulfur atoms have distorted tetra\u00adhedral geometries, with an O2\u2014S1\u2014C6 bond angle of 109.18\u2005(8)\u00b0. The two naphthalene units in each dansyl group are inclined to one another at an angle of 52.29\u2005(6)\u00b0; however, no stacking of the naphthalene units is observed.The title compound crystallizes in the space group le Fig.\u00a01. The hydc-axis direction, Fig.\u00a02B\u2014O3 and C16\u2014H16C\u2014O2 contacts further link the mol\u00adecules into sheets in the ab plane, Fig.\u00a03a-axis direction, Fig.\u00a04In the crystal structure, the supra\u00admolecular packing is dominated by weak C\u2014H\u22efO hydrogen bonds, Table\u00a01GAUSSIAN09 program package calculations were performed at the CAM-B3LYP/6-311G level as implemented in the et al., 2016et al., 2015N-cyclo\u00addodecyl-5-(di\u00admethyl\u00adamino)\u00adnaphthalene-1-sulfonamide -5-(di\u00admethyl\u00adamino)\u00adnaphthalene-1-sulfonamide di \u00adnaphthalene-1-sulfonamido]\u00adphenyl 5-(di\u00admethyl\u00adamino)\u00adnaphthalene-1-sulfonate arene complexes [5-(di\u00admethyl\u00adamino)\u00adnaphthalene-1-sulfonamido]\u00adbis\u00ad(tri\u00adphenyl\u00adphosphine)digold (UZEJAL) and [5-(di\u00admethyl\u00adamino)\u00adnaphthalene-1-sulfonamido]\u00adtris\u00ad(tri\u00adphenyl\u00adphosphine)trigold perchlorate (UZEJEP) (Cho 2 atmosphere for 24\u2005h. The solvent was removed with a rotary evaporator. The residue was added to water (15\u2005ml) and extracted with di\u00adchloro\u00admethane (3 \u00d7 25ml). The organic layer was dried with anhydrous Na2SO4 and the product was purified by column chromatography using CH2Cl2 as the eluent. The di\u00adchloro\u00admethane was slowly evaporated to afford a green solid in 65% yield. Light-green block-like crystals were grown in chloro\u00adform at room temperature.The title compound was synthesized by mixing 3,5-di\u00adhydroxy\u00adtoluene and dansyl chloride using potassium carbonate as a base in aceto\u00adnitrile solvent (40\u2005ml). The reaction mixture was heated at 363\u2005K and stirred under an Nd(C\u2014H) = 0.95\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for aromatic and d(C\u2014H) = 0.98\u2005\u00c5, Uiso(H) = 1.5Ueq(C) for methyl H atoms. As atom Cl lies on a twofold rotation axis, the H atoms of the Cl methyl group are disordered with occupancies fixed at 0.5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019009058/sj5573sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019009058/sj5573Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019009058/sj5573Isup3.cmlSupporting information file. DOI: 1535824CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions lead to the formation of a two-dimensional network structure parallel to (110). Hirshfeld surface analysis confirmed the inter\u00admolecular inter\u00adactions.The title compound was synthesized from naphthalene di\u00adimide and methyl\u00adthio\u00adpropi\u00adamine. The asymmetric unit consists of half of the total mol\u00adecule as the mol\u00adecule lies on an inversion center. Intra\u00admolecular C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds cause the mol\u00adecule to have an 22H22N2O4S2, was synthesized by the reaction of 1,4,5,8-naphthalene\u00adtetra\u00adcarb\u00adoxy\u00adlic dianhydride with 3-(methyl\u00adsulfan\u00adyl)propyl\u00adamine. The whole mol\u00adecule is generated by an inversion operation of the asymmetric unit. This mol\u00adecule has an anti form with the terminal methyl\u00adthio\u00adpropyl groups above and below the aromatic di\u00adimide plane, where four intra\u00admolecular C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds are present and the O\u22efH\u22efS angle is 100.8\u00b0. DFT calculations revealed slight differences between the solid state and gas phase structures. In the crystal, C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds link the mol\u00adecules into chains along the [2ABAB sequence along the c-axis direction. Hirshfeld surface analysis shows that van der Waals inter\u00adactions make important contributions to the inter\u00admolecular contacts. The most important contacts found in the Hirshfeld surface analysis are H\u22efH (44.2%), H\u22efO/O\u22efH (18.2%), H\u22efC/C\u22efH (14.4%), and H\u22efS/S\u22efH (10.2%).The title compound, C The mols Table\u00a01. The terGAUSSIAN09 software package \u2005\u00c5; Cg1 and Cg2 are the centroids of the C6/C7/C7iii/C8iii/C10/C11iii and C6iv/C7iv/C7v/C8v/C10iv/C11v rings, respectively; symmetry codes: (iii) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z; (iv) \u2212x\u00a0+\u00a02, \u2212y, \u2212z; (v) x, y\u00a0\u2212\u00a01, z]. These \u03c0\u2013\u03c0 inter\u00adactions lead to a two-dimensional network structure parallel to the (001) plane indicate the Hirshfeld surface analysis was performed using a\u2013e), which show the H\u22efH, H\u22efC/C\u22efH, H\u22efO/ O\u22efH, H\u22efN/N\u22efH, and H\u22efS/S\u22efH contacts. The relative contributions of the atomic contacts to the Hirshfeld surface are summarized in Table\u00a03The C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds and \u03c0\u2013\u03c0 stacking inter\u00adactions are identified in the two-dimensional fingerprint plots Fig.\u00a07a\u2013e, whiA mixture of 1,4,5,8-naphthalene\u00adtetra\u00adcarb\u00adoxy\u00adlic dianhydride and 3-(methyl\u00adsulfan\u00adyl)propyl\u00adamine in toluene (5\u2005mL) and quinoline (15\u2005mL) was heated at 453\u2005K with stirring for 1h. Upon cooling to room temperature, a golden yellow crude solid was filtered off and washed with diethyl ether. A golden yellow powder was obtained. Crystals suitable for X-ray diffraction analysis were obtained by slow evaporation of a di\u00adchloro\u00admethane solution of the title compound.1H NMR : \u03b4 8.77 , 4.33 , 2.64 , 2.14 , 2.07 . 13C NMR : \u03b4 162.81, 130.99, 126.69, 126.57, 40.01, 31.61, 27.16 and 15.31. IR : 3344 (m); 3071 (m); 2916 (s); 2848 (s); 1999 (s); 1693 (s).et al., 2016et al., 2013n-alkyl groups are known with 2,7-di\u00adbutyl\u00adbenzo[lmn]phenanthroline-1,3,6,8-tetra\u00adone naphthalene-4,5,8,9-tetra\u00adcarb\u00adoxy\u00adlic acid di\u00adimide = 0.95\u2005\u00c5, Uiso = 1.2Ueq(C) for aromatic, d(C\u2014H) = 0.99\u2005\u00c5, Uiso = 1.2Ueq(C) for methyl\u00adene, and d(C\u2014H) = 0.98\u2005\u00c5, Uiso = 1.5Ueq(C) for the methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989019007771/sj5571sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989019007771/sj5571Isup2.hklStructure factors: contains datablock(s) I. DOI: 1919395CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III compounds is the octa\u00adhedral coordination of the IrIII atom by a PCP pincer complex, a C atom of a (eth\u00adoxy\u00adoxoethanyl\u00adidene)methane group and two variable ligands X and Y .The common structural feature of the four title Ir 4P,C,C\u2032,P\u2032)chlorido\u00adhydridoiridium(III) chloride methyl\u00adene chloride 2.75-solvate (4), (bis\u00ad{[(di\u00adphenyl\u00adphosphan\u00adyl)meth\u00adyl]di\u00adphenyl\u00adphosphanyl\u00adidene}(eth\u00adoxy\u00adoxoethanyl\u00adidene)methane-\u03ba4P,C,C\u2032,P\u2032)chlorido\u00ad(eth\u00adoxy\u00adoxoethanido)iridium(III) chloride\u2013methanol\u2013water (1/1/0.5) (5), (bis\u00ad{[(di\u00adphenyl\u00adphosphan\u00adyl)meth\u00adyl]di\u00adphenyl\u00adphosphanyl\u00adidene}(eth\u00adoxy\u00adoxoethanyl\u00adidene)methane-\u03ba4P,C,C\u2032,P\u2032)di\u00adchlorido\u00adiridium(III) chloride\u2013methanol\u2013water (1/1/2) (6) and (bis\u00ad{[(di\u00adphenyl\u00adphosphan\u00adyl)meth\u00adyl]di\u00adphenyl\u00adphosphanyl\u00adidene}(eth\u00adoxy\u00adoxoethanyl\u00adidene)methane-\u03ba4P,C,C\u2032,P\u2032)carbon\u00adyl(eth\u00adoxy\u00adoxoethanide)iridium(III) dichloride\u2013meth\u00adyl\u00adene chloride\u2013water (1/2/1.5) (7) or in terms of their formulae [Ir(C55H50O2P4)ClH]Cl\u00b72.75CH2Cl2 (4), [Ir(C4H7O2)(C55H50O2P4)Cl]Cl\u00b7CH3OH\u00b70.5H2O (5), [Ir(C55H50O2P4)Cl2]Cl\u00b7CH3OH\u00b72H2O (6) and [Ir(C4H7O2)(C55H50O2P4)(CO)]Cl2\u00b72CH2Cl2\u00b71.5H2O (7) is a central IrIII atom coordin\u00adated in a distorted octa\u00adhedral fashion by a PCCP ligand system and two additional residues, such as chlorides, a hydride, a carbonyl or an alkyl unit. Thereby, the PCP pincer ligand system and the residue trans to the carbodi\u00adphospho\u00adrane (CDP) C atom surround the iridium(III) transition metal in the equatorial plane under the formation of two five-membered dissimilar chelate rings . A cyclo\u00adpropane-like heterocycle is positioned approximately orthogonal (84.21\u201388.85\u00b0) to the equatorial plane, including an alkyl\u00adidene bridge connecting the IrIII atom and the coordinating CDP atom of the PCP subunit. In general, the neutral PCCP ligand system coordinates the metal in a tetra\u00addentate way via three Lewis acid/base bonds and by an alkyl\u00adidene unit presenting strengthened inter\u00adactions. In all the crystal structures, (disordered) solvent mol\u00adecules are present in the voids of the packed mol\u00adecules that inter\u00adact with the positively charged complex and its chloride counter-ion(s) through weak hydrogen bonding.The common feature of the four iridium(III) salt complexes, (bis\u00ad{[(di\u00adphenyl\u00adphosphan\u00adyl)meth\u00adyl]di\u00adphenyl\u00adphosphanyl\u00adidene}(eth\u00adoxy\u00adoxoethanyl\u00adidene)methane-\u03ba Nomura et al., 2011et al., 1998et al., 1991et al., 2005Carbodi\u00adphospho\u00adranes (CDP) in combination with transition metals initialize a huge variety of functionalities. As a result of the presence of two 2]Cl atom, followed by deprotonation of the carbodi\u00adphospho\u00adrane carbon atom, the generation of a hydrido ligand caused by an oxidation of the iridium(I) atom and the formation of the Cl complex 1 an Cl2 precursor system (2) is generated in high yields (86%). Moreover, the preparation of complex 2 in a less polar solvent environment like chloro\u00adform/aceto\u00adnitrile or in a solvent mixture of methyl\u00adene chloride/aceto\u00adnitrile (v/v 5:1) is not possible and qu\u00adanti\u00adtatively results in the substitution of one phosphine moiety of the carbodi\u00adphospho\u00adrane functionality against the carbene CHCO2Et subunit. An Cl complex 3 is generated, offering a phospho\u00adrus ylide carbon backbone to 333\u2005K for 2\u2005h benefits the ring-opening reaction of the PCCP pincer ligand system. Therefore, a reorganization of the ligand system is supported, resulting in the qu\u00adanti\u00adtative formation of complex 3. Furthermore, evaporation of the reaction mixture of complex 2 causes an exchange of the aceto\u00adnitrile solvent ligand with a chloride counter-ion and the creation of the desired Cl complex 4. If the starting materials Cl alkyl derivative 5. This reaction procedure is well known, and the mechanism of the inter\u00admolecular insertion reaction has been clarified via an inter\u00admediate carbene complex (dppm)2-\u03ba4P,C,C\u2032,P\u2032}Cl2]Cl complex 6. Besides, a replacement of the chlorido ligand of compound 5 by a carbonyl group is possible and results in the Cl2 complex 7.The stucture of this irid\u00adium(III) PCCP complex was completely determined by NMR spectroscopy and X-ray crystallography, but up to now crystallization attempts of the inter\u00admediates, 4\u20137.Here we report details of the syntheses and crystal structures of complexes 4, Cl, comprises of one formula unit of 4 and additionally of 2.75 mol\u00adecules of methyl\u00adene chloride solvent mol\u00adecules. The central iridium(III) transition metal is surrounded in a distorted octa\u00adhedral fashion by a PCCP pincer-like ligand system, and anionic chlorido and hydrido ligands (dppm)2-\u03ba4P,C,C\u2032,P\u2032] ligand coordinates the IrIII metal in a tetra\u00addentate fashion via two P and two C atoms under formation of two five-membered, dissimilar chelate rings and one three membered heterocycle. The PCP ligand exhibits a meridional arrangement with the hydrido ligand completing the equatorial plane trans to the C1 carbodi\u00adphospho\u00adrane atom. A cyclo\u00adpropane-like chelate ring is positioned nearly normal (84.21\u00b0) to the equatorial plane, and a chlorido ligand is positioned trans to the alkyl\u00adidene carbon atom C4. The Ir\u2014C1 [2.273\u2005(4)\u2005\u00c5] and Ir\u2014C4 [2.072\u2005(5)\u2005\u00c5] distances differ significantly and consequently these values substanti\u00adate a strengthened inter\u00adaction between the iridium(III) metal and the alkyl\u00adidene carbon atom. The C1\u2014C4 separation [1.515\u2005(6)\u2005\u00c5] is slightly shorter in comparison to a typical C\u2014C single bond but, in general, very close to that of cyclo\u00adpropanes. However, in comparison with a cyclo\u00adpropane mol\u00adecule the C4\u2014Ir1\u2014C1 [40.5\u2005(2)\u00b0], C4\u2014C1\u2014Ir1 [62.6\u2005(2)\u00b0] and C1\u2014C4\u2014Ir1 [76.9\u2005(3)\u00b0] angles emphasise a significant distortion of the synthesized three-membered heterocycle. All mentioned geometric features of this strained Ir\u2014C1\u2014C4 metallacycle can be associated with the structural results of the Ru\u2014C\u2014C triangle reported by Zhang et al. (dppm)2-\u03ba4P,C,C\u2032,P\u2032}(CH2CO2Et)Cl]Cl, is defined by one complex 5, one half-occupied water mol\u00adecule and one disordered methanol solvent mol\u00adecule. In comparison with the structural features discussed in detail for compound 4, significant differences pertain only to the equatorial position trans to C1. Here the hydrido ligand in 4 is exchanged by an ethyl acetate unit (dppm)2-\u03ba4P,C,C\u2032,P\u2032}Cl2]Cl. In its crystalline form, besides one formula unit of 6, one solvent mol\u00adecule of MeOH and two water mol\u00adecules in total are present in the asymmetric unit. Overall, this PCCP derivative shows very similar structural characteristics (dppm)2-\u03ba4P,C,C\u2032,P\u2032}(CH2CO2Et)(CO)]Cl2 = 2.83\u2005\u00c5] exhibiting distances shorter than the sum of the van der Waals radii ], 2.91\u2005\u00c5 (H3A\u22efCl2) and 2.40\u2005\u00c5 (H3B\u22efO1) Fig.\u00a07.6, the methyl\u00adene groups of the PCP unit and the chloride counter-ion and the solvent mol\u00adecules form C\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions ], 2.66\u2005\u00c5 (H3B\u22efCl3) and 2.40\u2005\u00c5 [H6A\u22efO3 ] ] Fig.\u00a08.7, the chloride counter-ions inter\u00adact with both the PCP pincer ligand system and the solvent mol\u00adecules. The solvent mol\u00adecules also show inter\u00adactions with the iridium complex III{C(CHCO2Et)(dppm)2-Synthesis of Cl\u00b72.75CH2Cl2 (4)\u03ba: A mixture of [CH(dppm)2]Cl and [IrCl(cod)]2 was solved in 0.1\u2005ml of MeCN. After a reaction time of one minute, a solution of ethyl diazo\u00adacetate in MeOH (0.5\u2005ml) was added. 10\u2005min later, a deep yellow liquid was obtained. The volatiles were removed and the remaining solid was dissolved in methyl\u00adene chloride (0.6\u2005ml), leading to complex 4 in high yield . Single crystals of complex 4 were grown from a solvent mixture of n-hexane (1.2\u2005ml) and CH2Cl2 (0.2\u2005ml). 31P {1H} NMR (CHCl3): \u03b4 = 18.8 , 38.1 , 34.7 , 10.7 ppm; 1H NMR : \u03b4 = \u221215.2 ppm; 13C {1H} NMR (CDCl3): \u03b4 = 3.6 ppm.III{C(CHCO2Et)(dppm)2-Synthesis of Cl\u00b7CH3OH\u00b70.5 H2O (5):} was added to a solution of complex 4 in CH2Cl2 (0.6\u2005ml), and the reaction mixture was stirred for 30\u2005min. Complex 5 was formed qu\u00adanti\u00adtatively. Single crystals were obtained via slow evaporation of a 1:1methyl\u00adene chloride/methanol mixture. 31P {1H} NMR (CHCl3): \u03b4 = 0.3 , 40.6 , 36.4 , \u22124.4 ppm; 13C {1H} NMR (CDCl3): \u03b4 = 3.1 ppm.III{C(CHCO2Et)(dppm)2-Synthesis of Cl\u00b7CH3OH\u00b72H2O (6):}Cl A solution of complex 4 in CH2Cl2 (0.6\u2005ml) was treated with hydro\u00adchloric acid and stirred vigorously for approximately 10\u2005min. The organic phase was separated and washed with water (0.5\u2005ml) three times in total. Complex 6 was formed almost qu\u00adanti\u00adtatively. Yellow single crystals were generated by slow evaporation of a 1:1 solvent mixture of MeCN and MeOH. 31P {1H} NMR (CHCl3): \u03b4 = \u22126.1 , 46.9 , 45.8 , \u221210.3 ppm; 13C {1H} NMR (CDCl3): \u03b4 = 3.8 ppm.III{C(CHCO2Et)(dppm)2-Synthesis of Cl2\u00b72CH2Cl2\u00b71.5H2O (7):} in CH2Cl2 was placed under an atmosphere of CO. After a reaction time of 1\u2005h, complex 7 had formed qu\u00adanti\u00adtatively . Single crystals were grown from a solution of methyl\u00adene chloride, covered with a small amount of ethyl acetate. 31P {1H} NMR (CH2Cl2): \u03b4 = \u22126.5 , 41.2 , 39.7 , \u221216.4 ppm; 13C {1H} NMR (CD2Cl2): \u03b4 = 16.1 , 172.8 ppm.Crystal data, data collection and structure refinement details are summarized in Table\u00a06Uiso(H) = 1.2Ueq(C) for phenyl and methyl\u00adene H atoms and 1.5Ueq(C) for methyl H atoms.Unless noted otherwise, H atoms in the four structures were placed geometrically and refined in the riding-model approximation with 4, the two hydrogen atoms bound to the central Ir1 atom and the C4 atom of the eth\u00adoxy\u00adoxoethanyl\u00adidene moiety were discernible from a difference-Fourier map. They were refined with bond-length restraints of 0.96\u2005\u00c5 (C4) and 1.60\u2005\u00c5 (Ir1) and with individual Uiso values. Three of the four methyl\u00adene chloride solvent mol\u00adecules are disordered. One solvent mol\u00adecule shows half-occupation, one is disordered around an inversion centre (occupancy 0.25) and for one the Cl atoms show a positional disorder over two sites (ratio 0.7:0.3). All H atoms of the solvent mol\u00adecules were omitted from the final model.For compound 5 was poor. Hence, it was possible to collect reflections only up to 45\u00b0/2\u03b8. The H atom attached to the C4 position was treated as described above. The methanol and water (O7) solvent mol\u00adecules are disordered around an inversion centre and were refined with half-occupation. H atoms of the disordered solvent mol\u00adecules were omitted from the model. Furthermore, one phenyl group shows a 1:1 positional disorder and was refined over two sets of sites . All atoms of the disordered phenyl ring were refined isotropically.The scattering power of the crystal of compound 6 and 7, the H atom attached to the C4 position was treated as described above. For 6, localization of the H atoms of the methanol and water solvent mol\u00adecules was not possible and hence they were omitted from the model. For 7, H atoms of water mol\u00adecule O6 were located from a difference-Fourier map and refined with bond-length restraints of 0.84\u2005\u00c5. The O7 atom of the other water mol\u00adecule was treated as being half-occupied, and its H atoms were omitted from the model. One methyl\u00adene chloride solvent mol\u00adecule was refined over two sets of sites (ratio 0.65:0.35).In compounds 10.1107/S2056989018017024/wm5471sup1.cifCrystal structure: contains datablock(s) global, 4, 5, 6, 7. DOI: 10.1107/S2056989018017024/wm54714sup6.hklStructure factors: contains datablock(s) 4. DOI: 10.1107/S2056989018017024/wm54715sup7.hklStructure factors: contains datablock(s) 5. DOI: 10.1107/S2056989018017024/wm54716sup8.hklStructure factors: contains datablock(s) 6. DOI: 10.1107/S2056989018017024/wm54717sup9.hklStructure factors: contains datablock(s) 7. DOI: 1873392, 1873391, 1873390, 1873389CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "This is comparable to very weak CH hydrogen bonding interactions and is in line with the weak interaction energies calculated of typical charge neutral adducts such as [Me3N-CH3\u00b7\u00b7\u00b7OH2] (2a). The interaction energy is enhanced to \u2264\u20135 kcal\u00b7mol\u22121 when X is more electron withdrawing such as in [O2N-CH3\u00b7\u00b7O=Cdme] (20b) and to \u226418 kcal\u00b7mol\u22121 in cationic species like [Me3O+-CH3\u00b7\u00b7\u00b7OH2]+ (8a).A systematic evaluation of the CSD and the PDB in conjunction with DFT calculations reveal that non-covalent Carbon-bonding interactions with X\u2013CH The manner in which molecules interact with one another is largely determined by non-covalent interactions.c So-calle3-hybridized C-atoms. Such interactions have been studied since about 2013 [3\u00b7\u00b7\u00b7OH2] and [HO-3\u00b7\u00b7\u00b7OH2] of \u22120.7 6\u201311a were computed as well and the most stable of these involved the Me3C+ carbocation in 6 (adducts with pentamethylated Carbon are unstable). The bonding interaction in 6b is largely covalent, as evidenced by the interaction energy of \u221282.4 kcal\u00b7mol\u22121, the large orbital contribution (55%), a dense bond critical point (18.4 \u00b7 102 a.u.) and a clear pyramidalization of the central C-atom (see 6a (Me3C+\u00b7\u00b7\u00b7O interaction) and 9 (with the least electronegative P) converged into an X\u2013CH3\u00b7\u00b7\u00b7O Carbon bonding geometry. This is illustrated for 8a and 11a in methylC and Owater with a bond density of 1.15 \u00b7 102 and 0.98 \u00b7 102 a.u. for 8a and 11a respectively. The bonding energies in 7\u201311a are mainly electrostatic in origin (~70%) and about \u22128 kcal\u00b7mol\u22121 for water and \u221215 kcal\u00b7mol\u22121 for dma. The most stable adducts in both series involved the most electronegative O (3.44) in Me3O+ (8). Two alternative configurations with N-methylpyridinium were also computed (12 and 13). In 12, the O points in between two CH hydrogens as is illustrated for 12a in 13 the O atom is located directly above the cationic N+. Both 12 and 13 are more stable than the Carbon bonding geometry found in 11, suggesting that hydrogen bonding interactions are most preferred. The interaction energies of Carbon bonding interaction with cationic species is similar to previous data of the adducts: [H3N+-CH3\u00b7\u00b7\u00b7OCH2] ; [3N+-CH3\u00b7\u00b7\u00b7OC(H)NH2] ; [2S+-CH3\u00b7\u00b7\u00b7OH2/NH3/OCH2] ; [2S+-CH3\u00b7\u00b7\u00b7various lone-pairs] ; [3\u00b7\u00b7\u00b7C2H2] ; [3\u00b7\u00b7\u00b7C2H4/NH3/PH3] ; [3\u00b7\u00b7\u00b7C2H4/dma] ; [2N-CH3\u00b7\u00b7\u00b7dma] [As the calculations with cationic species imply that electron withdrawing substituents amplify the Carbon bonding interaction, it was decided to compute adducts with small molecules that have an electron withdrawing group: Iodomethane (\u00b7mol\u22121); ,51,52,53\u00b7mol\u22121); [Hlg-CH3\u00b7mol\u22121); [NC-CH3\u00b7\u00b7mol\u22121); ,56 and [2-O in dma (adducts \u2018b\u2019) is consistently about twice as strong as the interaction with sp3-O in water (adducts \u2018a\u2019). This is in line with the larger amount of van der Waals overlap observed in the N(d\u2019) plots , to moderately weak to fairly strong in the cationic adducts . \u0394E becomes smaller (more stable) in the order 2 < 14 < 16 < 18 < 20 < 7 < 8. Within this series, the orbital contribution remains constant at about 15\u201320%, while the electrostatic component increases from 30\u201335% in 2 to about 65% in 8. This implies that stronger Carbon bonding interactions are mainly driven by electrostatic interactions and that weaker such adducts are driven by dispersion. These computational results are consistent with recent literature reports [3 is strongly polarized within a hemisphere of 5 \u00c5 basal radius (centered on C). It was found that X\u2013CH3\u00b7\u00b7\u00b7ElR interactions can be as directional as very weak hydrogen bonding interaction involving C\u2013H (Pmax \u2264 1.50) but not directional at all when X = C. Grouping of data with significant amounts of van der Waals overlap (up to ~30%) was observed in various sub-datasets in the region where the X\u2013CH3\u00b7\u00b7\u00b7ElR angle \u03b1 is 160\u00b0\u2013180\u00b0. These distributions were significantly shifted to shorter distances in the case of cationic R3N+\u2013CH3\u00b7\u00b7\u00b7Owater/amide compared to charge-neutral R2N\u2013CH3\u00b7\u00b7\u00b7Owater/amide interactions.The CSD and the PDB were systematically evaluated for potential directional behavior of intermolecular non-covalent Carbon bonding interactions involving X\u2013CH3\u00b7\u00b7\u00b7O adducts with water and dimethylacetamide are very weak and are often not the energy minima of the adducts . The interaction energies can be increased by deploying a more electron withdrawing X . Rendering X cationic leads to even more stable adducts in 7, 8, 10 and 11). Carbon-bonding adducts with dimethylacetamide are consistently twice as stable as those with water. Energy decomposition analyses showed that increased stability is driven by electrostatics and atom-in-molecule analyses regularly gave a clear bond critical point involving the methyl C-atom.Model DFT calculations revealed that charge neutral X\u2013CH3 is electrostatically driven and can be significant. The interaction can even by mildly directional in the solid state (comparable to weak CH hydrogen bonding interactions), provided X is sufficiently electron withdrawing.It is thus concluded that this combined database / DFT study reaffirms that intermolecular non-covalent Carbon interactions with X\u2013CH"} +{"text": "I) and 1,2\u2010disila[9]crown\u20103 (II) with MgI2 yielded exclusively [MgI2] (1). The larger Ca2+ ion was then employed for cross\u2010coupling of I and II and yielded the complex [CaI2] (2). Cross\u2010coupling of I and 1,2,4,5\u2010tetrasila[9]crown\u20103 (III) with SrI2 enables the synthesis of the silicon\u2010dominant 1,2,4,5,10,11\u2010hexasila[15]crown\u20105 ether complex of SrI2 (3). Further, the compounds [SrI2] (4), [SrI2] (5), and [SrI2] (6) were obtained by coupling I, 1,2\u2010disila[12]crown\u20104 (IV) or 1,2\u2010disila\u2010benzo[12]crown\u20104 (V), respectively. Using various anions, the (cross\u2010)coupled ligands were also observed in an X\u2010ray structure within the mentioned complexes. These template\u2010assisted (cross\u2010)couplings of various ligands are the first of their kind and a novel method to obtain macrocycles and/or their metal complexes to be established. Further, the Si\u2212O bond activations presented herein might be of importance for silane or even organic functionalization.Alkaline earth metal iodides were used as templates for the synthesis of novel silicon\u2010based ligands. Siloxane moieties were (cross\u2010)coupled and ion\u2010specific, silicon\u2010rich crown ether analogues were obtained. The reaction of 1,2,7,8\u2010tetrasila[12]crown\u20104 coupled and ion specific, silicon\u2010rich crown ether analogues were obtained. The coupling of various ligands gives an outlook on Group\u20052 ion\u2010catalyzed macrocycle and/or silane syntheses. It is well known for crown ethers to form coordination compounds with metal centers across a wide range of the periodic table. These complexes are generally very stable and these polyethers as well as related systems such as cryptands, or in general, multidentate ligands, gained many fields of applications since their discovery in the mid\u2010sixties. The synthesis of such polyethers, however, is often not trivial because the molecules of the starting materials need to be brought into a suitable conformation for the formation of a specific (most often cyclic) product. The synthesis of these cyclic ethers is therefore mostly metal\u2010template assisted.Templates are known to provide a suitable coordination sphere for the starting materials, which favors the linkage to ring closure and allows obtaining cyclic ligand systems in much higher yields.\u03b4+ and metaln+ disturbs silyl ether bonding.n type .+ and Cs+ so far relucted silyl ether coordination.Most recently, both concepts\u2014covalency and ionicity\u2014were considered as harmonious models to understand the basicity of siloxanes. Regarding a covalent model, negative hyperconjugation interactions are described in the case p(O)\u2192\u03c3*(Si\u2212C).Apart from siloxane coordination chemistry, the metal\u22c5\u22c5\u22c5O\u2212Si interaction should also be put in context to Si\u2212O bond activation. From the perspective of Si\u2212O bond activation, weakening of Si\u2212O bonds is of considerable significance regarding the conversion of silicates to silanes. Especially dicatecholato\u2010silicate(IV) complexes are frequently used as precursors for either silane synthesis or functionalization of various organic molecules.2+ ion, we aimed at synthesizing a [15]crown\u20105 ether by cross\u2010coupling the ligands 1,2,7,8\u2010tetrasila[12]crown\u20104 (I) and 1,2\u2010disila[9]crown\u20103 (II). However, the reaction in \u03b1,\u03b1,\u03b1\u2010trifluorotoluene heated at reflux failed (Scheme\u20052] (1) was obtained and, as can be seen from the molecular structure determined by single\u2010crystal X\u2010ray diffraction analysis (SC\u2010XRD), the moiety of crown ether I is large enough to bind Mg2+ I2] and the octahedral coordination towards Mg2+ is preferred.I with Mg2+ is present because the 29Si{1H}\u2005NMR chemical shift is observed at 22.8\u2005ppm and thus, a strong lowfield shift in comparison with I (\u03b4=10.8\u2005ppm)Starting with the Mgsizing a 5crown\u20105 2+ ion was described earlier and, as can be seen from various examples, seems to match perfectly.I and II with CaI2. After workup procedures two new signals in the 29Si{1H}\u2005NMR are observed at 19.6 and 19.9\u2005ppm. Crystallization of the reaction product 2 however, was unsuccessful from different solvents and temperatures also due to poor solubility. We therefore expanded the anion I\u2212 to I3\u2212 upon iodine addition. The compound [Ca(I3)2] (2\u2009a) then was crystallized and the molecular structure was determined by means of SC\u2010XRD and 245.2(9)\u2005pm and compare well to the complex [CaOTf2] (OTf=CF3SO3\u2212).The fivefold, coplanar coordination by sila\u2010crown ligands towards the larger Ca2] (3) is another example of a cross\u2010coupling reaction in which an alkaline earth metal cation is used. 3 represents the first hybrid disila\u2010crown ether bearing more disilane than ethylene units. The even larger Sr2+ cation can be used as a convex template to cross\u2010couple I and 1,2,4,5\u2010tetrasila[9]crown\u20103 (III), which is accessible applying reaction conditions reported before.I and one equivalent of III. Due to three disilane units within the ligand framework, the Sr2+ cation fits well in the cavity of the obtained ligand. With 19.44, 19.27, and 13.32\u2005ppm, three resonances are observed in the 29Si{1H}\u2005NMR spectrum, all of which compare well to those of OSi\u22c5\u22c5\u22c5Sr2+ coordination compounds characterized before.3 could not be obtained, even after iodine addition. Hence, the Lewis acidic salt GaI3 was added as an acceptor for the iodide anion. It is notable, that also in 3\u2009a, the SiSi\u2212O\u2212SiSi fragment of the siloxane framework provides a small Si\u2010O\u2010Si angle, which measures 121.1(6)\u00b0. The OSi\u2212O\u2212Si\u22c5\u22c5\u22c5Sr2+ distance might be the longest O\u22c5\u22c5\u22c5Sr2+ distance observed in 3\u2009a, but the high basicity of siloxanes towards Group\u20052 ions is clearly emphasized.The synthesis of [SrI4)2] (3\u2009a) were obtained, which were analyzed through SC\u2010XRD compare well to various complexes of Sr2+ and sila ligands, which were reported before.2+ cation as a template, we were able to observe reactions of other disila\u2010crown ethers as well. The small cavity of II does not allow for 1:1 complexation of Sr2+ and thus, a total of two equivalents of II react to form the 1,2,10,11\u2010tetrasila[18]crown\u20106 ether. [SrI2] (4) is obtained as a colorless powder and in solution, a resonance at 17.5\u2005ppm in the 29Si{1H}\u2005NMR spectrum was observed. Given that we were also experiencing problems with the crystallization of this compound, the X\u2010Ray structure could only be determined upon iodine addition. According to a reduced niggli formula, compound 1\u221e+ fragments to build infinite chains along [001] which is most likely the driving force for crystallization. The OSi\u22c5\u22c5\u22c5Sr2+ distances in 4\u2009a compare well to those in compound 3\u2009a and [SrI2].After several months, a few single crystals of [Sr(GaI4)2] \u2009a were o4 as an example. As drawn here, the sila\u2010ligand is polarized by the Lewis acid first, which makes it electrophilic and thus accessible for nucleophilic attack of I\u2212. Afterwards, the Si\u2212O bond is cleaved. Subsequently, the intermediate then has to cleave the second sila\u2010crown. At this point of the reaction, the alkaline earth metal ion acts as a convex template, in which the open\u2010chained ligand species is brought into a suitable conformation for ring closure, which is the final step of the presented reactions. We want to emphasize, that such a mechanism was also postulated by Harder in a past work.At this point, it is clear that (cross\u2010)coupling reactions are possible with different alkaline earth metal iodides. We tried to understand how the reaction works and the mechanism behind the formation of these unprecedented macrocycles, but unfortunately, we were unable to characterize any intermediate products by means of NMR spectroscopy or SC\u2010XRD. We can assure that heating at reflux is needed to cleave the smaller rings. Further, we can assure that the reaction does only occur if an iodide salt is employed. Other alkaline earth metal halides do not form these ligands. This lets us conclude that one key step in the formation of the (cross\u2010)coupling product has to be a nucleophilic substitution reaction. Furthermore, as evident from different works, we could convincingly show that the Si\u2212O bond is significantly weakened due to coordinating an alkaline earth metal ion. Thus, the Si\u2212O bond instead of the C\u2212O bond is cleaved, which is also represented by the herein obtained macrocycles. A proposed mechanism for the formation of the cross coupling products is depicted in Scheme\u20052 with the ligands 1,2\u2010disila[12]crown\u20104 (IV) and 1,2\u2010disila\u2010benzo[12]crown\u20104 (V) were performed. Both ligands are too small for Sr2+ and thus, template\u2010assisted ring opening yielded novel species by intermolecular coupling of the respective crown ether. The reaction of IV with SrI2 results in the formation of the first disilanyl\u2010bearing [24]crown\u20108 ether. [SrI2] (5) was obtained by coupling two equivalents of IV I2].29Si{1H}\u2005NMR resonance signal for 5 is observed at 15.2\u2005ppm. A split of this resonance at low temperature of 190\u2005K was not observed.Further reactions of SrIwn\u20108)I2] was obta5 decomposes readily with only traces of moisture forming [SrI]I (EO7=heptaethylene glycol).Thus, the exchange between the coordinating disilane units is too fast on the NMR time scale and results in the described equivalency. At this point, it should be noted that the spectroscopic investigation of the compound is challenging. Compound 6) is more stable and was characterized by state\u2010of\u2010the\u2010art methods including SC\u2010XRD I]I is more 6 was reacted with [222]cryptand and indeed we were able to obtain free ligand 1,2,13,14\u2010tetrasila\u2010dibenzo[24]crown\u20108 (7) after workup. The metal\u2010free ligand species was characterized by means of HR\u2010ESI MS as well as NMR spectroscopy. In HR\u2010ESI+ MS, m/z relations of 625.2502 [7+H]+ (100) as well as 647.2320 [7+Na]+ (100) were found. In addition, 29Si{1H}\u2005NMR spectroscopy revealed highfield chemical shift and a single resonance at 11.5\u2005ppm is found. This value compares well to the free ligand V and also to related metal\u2010free disila\u2010ligand systems.One of the remaining questions is of course whether it is possible to remove the metal center for possible use of such new ligand in coordination chemistry. We are currently investigating this subject, but are still at an early stage. To give an outlook, we performed exchange reactions. As an example, an excess of ted with 2cryptanda\u2010dibenzocrown\u20108 \u2010coupling reactions with BaI2+ ion is too small to cross\u2010couple small silicon\u2010based ligands, a cross\u2010coupling reaction was established with the larger Ca2+ and Sr2+ ions. Furthermore, the Sr2+ ion was used for coupling reactions of three different disila\u2010crown ethers. Barium as a template for coupling reaction, however, turned out to be unsuccessful. Although the organic [12]crown\u20104 is too small for the Mg2+ ion, the cavity of the disilane\u2010bearing analogue 1,2,7,8\u2010tetrasila[12]crown\u20104 (I) ether matches fine with the Mg2+ ion and the complex [MgI2] (1) was obtained. [CaI2] (2) and [SrI2] (3) were synthesized by cross\u2010coupling reaction of the small rings I and II for 2, and I and III for 3. The ligand in 3 represents the first crown ether with more disilane than ethylene units between the donor atoms. Finally, [SrI2] (4), [SrI2] (5) and [SrI]I (6) were characterized and obtained by template\u2010driven dimerization of II, IV, or V with SrI2. The ligand moieties observed in 2\u20136 cannot yet be synthesized by conventional silane chemistry and for this reason, the template\u2010assisted (cross\u2010)coupling of various ligands is an elegant way to obtain novel macrocycles and/or their metal complexes. Overall, this work gives an outlook on Group\u20052 ion\u2010catalyzed silane syntheses, especially due to the fact that first attempts showed that the metal ion can, in principle, be removed from a novel sila\u2010ether. We also hope that there will be a broader application of the presented Si\u2212O bond activations as a mean of molecule functionalization.In this study alkaline earth metal iodides were successfully used for the coupling reaction of different silicon\u2010based crown ether analogues to form novel ligand environments. The work presents Si\u2212O bond\u2010cleavage reactions driven by a macrocyclic effect due to metal\u2010template reaction of the respective silicon\u2010based ligands. Given that the Mg2 , CaI2 and SrI2 , BaI2 and GaI3 were finely ground and stored in a Mbraun glovebox under Ar. NMR spectra were recorded on a Bruker AV III HD 300\u2005MHz or AV III 500\u2005MHz spectrometer, respectively. Infrared (IR) spectra of the respective samples were measured using attenuated total reflectance (ATR) mode on the Bruker\u2010type spectrometer Alpha FT\u2010IR. ESI mass spectra were acquired with a LTQ\u2010FT Ultra mass spectrometer (Thermo Fischer Scientific) and LIFDI mass spectra were acquired on a JEOL AccuTOF\u2010GCv device. The resolution was set to 100.000. Elemental analysis was carried out on a Vario MicroCube. In case of complex\u20052 and 3, we were not able to obtain an accurate elemental analysis. This is probably due to the wax\u2010like, greasy characteristics of the compounds and/or formation of SiC during the measurement. The crown\u2010ethers I, II, IV, and V were synthesized according to literature\u2010known procedures.7 can only be obtained with major impurities of [222]crypt.All manipulations were carried out with rigorous exclusion of oxygen and moisture using basic Schlenk techniques establishing an inert\u2010gas atmosphere with a vacuum line. All solvents were dried and freshly distilled before use. The alkaline earth metal salts MgI3 were dissolved in THF (50\u2005mL). Subsequently, O(Si2Me4Cl)2n\u2010pentane (50\u2005mL) followed by filtration. Removing the solvent under reduced pressure yielded the crown ether as a colorless oil .Ethylene glycol and NEt1H\u2005NMR : \u03b4=0.18 2), 0.20 2), 3.69\u2005ppm ; 13C{1H}\u2005NMR : \u03b4=0.1 2), 2.8 2), 66.0\u2005ppm ; 29Si{1H}\u2005NMR : \u03b4=1.5 2), 11.4\u2005ppm 2). HR\u2010MS: ESI(+) m/z (%): 309.1196 [M+H]+ (100), 639.2143 [2\u2009m+Na]+ (100); IR: 2951 (m), 2896 (w), 2867 (w), 1457 (vw), 1388 (vw), 1247 (s), 1140 (m), 1089 (s), 1027 (s), 927 (m), 854 (m), 795 (s), 761 (vs.), 721 (m), 681 (m), 660 (m), 633 (m), 553 (m), 506 (vw), 487\u2005cm\u22121 (m).I was dissolved in \u03b1,\u03b1,\u03b1\u2010trifluorotoluene (10\u2005mL). Subsequently, MgI2 was added. Stirring the mixture at 60\u2009\u00b0C for 90\u2005min gave a white suspension. Removing the solvent under reduced pressure yielded a white precipitate which was extracted with DCM (10\u2005mL) followed by filtration. After washing with n\u2010pentane (4\u2005mL), and drying in vacuo, 1 was obtained as a colorless powder . For single\u2010crystal growth, the powder was dissolved in DCM (4\u2005mL) and layered with n\u2010pentane (20\u2005mL). Single crystals were obtained overnight as colorless needles.Compound 1H\u2005NMR : \u03b4=0.59 2), 4.23\u2005ppm ; 13C{1H}\u2005NMR : \u03b4=\u22120.46 2), 62.8\u2005ppm ; 29Si{1H}\u2005NMR : \u03b4=22.8\u2005ppm 2). MS: ESI(+) m/z (%): 503.0270 [M\u2212I]+ (15); IR: 2949 (vw), 2888 (vw), 1450 (w), 1398 (vw), 1369 (vw), 1247 (m), 1103 (m), 1056 (s), 1022 (s), 934 (vs.), 915 (s), 868 (m), 840 (m), 818 (s), 802 (s), 780 (vs.), 715 (s), 645 (m), 464\u2005cm\u22121 (m). CHN calcd for C12H32I2MgO4Si4\u22c5\u2009CH2Cl2 C, 21.81; H, 4.79. Found C, 21.66; H, 4.90.I together with II were dissolved in \u03b1,\u03b1,\u03b1\u2010trifluorotoluene (10\u2005mL). Subsequently, CaI2 was added. Heating the mixture at reflux for 90\u2005min resulted in a white suspension. Removing the solvent under reduced pressure yielded a white precipitate which was extracted with of DCM (10\u2005mL) followed by filtration. After washing with two portions of n\u2010pentane (4\u2005mL each), and drying in vacuo, 1 was obtained as a colorless powder . For single\u2010crystal growth, 2 and I2 were dissolved in DCM (4\u2005mL) and filtered. The filtrate was then layered with n\u2010pentane (20\u2005mL). A few brown platelets of 2\u2009a were obtained after more than two weeks at ambient temperature.Compound 1H\u2005NMR : \u03b4=0.47 , 0.48 , 3.95\u20134.03\u2005ppm ; 13C{1H}\u2005NMR : \u03b4=\u22120.7 , \u22120.7 , 63.2 , 64.8 , 71.2\u2005ppm ; 29Si{1H}\u2005NMR : \u03b4=19.6 , 19.9\u2005ppm . MS: ESI(+) m/z (%): 419.1521 [M\u2212CaI2+Na]+ (100), 397.1705 [M\u2212CaI2+H]+ (5). IR: 2947 (m), 2881 (w), 1455 (vw), 1400 (vw), 1248 (s), 1057 (s), 1029 (s), 943 (s), 795 (s), 768 (vs.), 723 (s), 635 (m), 505\u2005cm\u22121 (vw).I and III were dissolved in \u03b1,\u03b1,\u03b1\u2010trifluorotoluene (10\u2005mL). Subsequently, SrI2 was added. Heating the mixture at reflux for 90\u2005min resulted in a white suspension. Removing the solvent under reduced pressure yielded a white precipitate which was extracted with DCM (10\u2005mL), followed by filtration. After washing with two portions of n\u2010pentane (4\u2005mL each) and drying in vacuo, 3 was obtained as a colorless, greasy solid . For single\u2010crystal growth, product 3 and GaI3 were dissolved in \u03b1,\u03b1,\u03b1\u2010trifluorotoluene (5\u2005mL). The suspension was stirred 30\u2005min and gently warmed to 60\u2009\u00b0C for 5\u2005min. The mixture was then freed of the solvent, extracted with DCM (3\u2005mL) and filtered. The filtrate was then concentrated until the saturation point was reached and stored at \u221232\u2009\u00b0C. A few colorless platelets of 3\u2009a were obtained after more than four weeks.Compounds 1H\u2005NMR : \u03b4=0.44 2), 0.47 2), 0.56 2), 3.96\u2005ppm ; 13C{1H}\u2005NMR \u03b4=\u22120.5 2), \u22120.5 2), 4.1 2), 65.3 , 65.4\u2005ppm ; 29Si{1H}\u2005NMR : \u03b4=13.3 2), 19.3 2), 19.4\u2005ppm 2). MS: ESI(+) m/z (%): 698.9902 [M\u2212I]+ (100), 507.1697 [M\u2212SrI2+Na]+ (30). IR: 2960 (w), 2879 (vw), 1456 (vw), 1400 (vw), 1369 (vw), 1258 (s), 1074 (s), 1024 (s), 951 (s), 894 (m), 851 (m), 794 (vs.), 769 (vs.), 712 (s), 634 (m), 557 (m), 497 (w), 448\u2005cm\u22121 (w).II was dissolved in \u03b1,\u03b1,\u03b1\u2010trifluorotoluene (10\u2005mL). Subsequently SrI2 was added. Heating the mixture at reflux for 60\u2005min. resulted in a white suspension. Removing the solvent under reduced pressure gave a white precipitate which was extracted with DCM (20\u2005mL) followed by filtration. Upon washing with n\u2010pentane (5\u2005mL) and removal of the solvent, 4 was obtained as a pale white powder . For single crystal growth, product 4 (30\u2005mg 0.04\u2005mmol) and I2 were dissolved in DCM (4\u2005mL). Layering the solution with n\u2010pentane (20\u2005mL) yielded single crystals of 4\u2009a as brown blocks after three days.Compound 1H\u2005NMR : \u03b4=0.45 , 3.88 , 4.08\u2005ppm ; 13C{1H}\u2005NMR : \u03b4=0.1 , 63.0 , 72.8\u2005ppm ; 29Si{1H}\u2005NMR : \u03b4=17.5\u2005ppm . MS: ESI(+) m/z (%): 264.0475 [M\u22122I]2+ (93), 447.2058 [M\u2212SrI2+Li]+ (3), 463.1797 [M\u2212SrI2+Na]+ (55), 479.1537 [M\u2212SrI2+K]+ (15), 655.0003 [M\u2212I]+ (16). IR: 2943 (w), 2880 (w), 1459 (w), 1398 (vw), 1353 (w), 1247 (m), 1069 (s), 1039 (s), 944 (s), 927 (s), 861 (s), 840 (s), 819 (s), 795 (s), 768 (vs.), 718 (s), 629 (m), 546 (w), 523\u2005cm\u22121 (w). CHN calcd for C16H40I2O6Si4Sr: C, 24.57; H, 5.15. Found: C, 25.42; H, 5.35.IV was dissolved in \u03b1,\u03b1,\u03b1\u2010trifluorotoluene (15\u2005mL). Subsequently SrI2 was added. Heating the mixture at reflux for three hours resulted in a white suspension. Removing the solvent under reduced pressure yielded a white precipitate which was extracted with DCM (20\u2005mL), followed by filtration. The product 5 was then obtained as a colorless powder after removal of the solvent . For single\u2010crystal growth, the powder was dissolved in DCM (4\u2005mL) and layered with n\u2010pentane (20\u2005mL). Single crystals were obtained overnight as colorless needles.Compound 1H\u2005NMR : \u03b4=0.37 2), 3.94 , 3.95\u2005ppm ; 13C{1H}\u2005NMR : \u03b4=\u22120.09 , 62.8 , 70.5 , 73.5\u2005ppm ; 29Si{1H}\u2005NMR : \u03b4=15.2\u2005ppm . MS: ESI(+) m/z (%): 551.2311 [M\u2212SrI2+Na]+ (82); IR: 2961 (m), 2943 (m), 2882 (w), 1454 (w), 1396 (w), 1366 (w), 1349 (w), 1302 (w), 1259 (s), 1091 (s), 1073 (s), 1039 (s), 1017 (s), 951 (s), 931 (s), 861 (m), 840 (m), 791 (vs.), 764 (s), 732 (s), 661 (s), 636 (m), 563 (w), 492 (w), 424\u2005cm\u22121 (w). CHN calcd for C20H48I2O8Si4Sr: C, 27.86; H, 5.67. Found: C, 27.60; H, 5.67.V was dissolved in \u03b1,\u03b1,\u03b1\u2010trifluorotoluene (15\u2005mL). Subsequently, SrI2 was added. Heating the mixture at reflux for three hours resulted in a white suspension. Removing the solvent under reduced pressure yielded a white precipitate which was extracted with DCM (20\u2005mL) followed by filtration. The product 6 was then obtained as a colorless powder after removal of the solvent . For single\u2010crystal growth, the powder was dissolved in DCM (3\u2005mL) and layered with n\u2010pentane (20\u2005mL). Single crystals were obtained overnight as colorless platelets.Compound 1H\u2005NMR : \u03b4=0.45 , 4.15 , 4.42 , 7.02\u20137.16\u2005ppm ; 13C{1H}\u2005NMR : \u03b4=0.0 , 62.4 , 72.6 , 116.1 , 124.9 , 147.4\u2005ppm ; 29Si{1H}\u2005NMR : \u03b4=19.8\u2005ppm . MS: ESI(+) m/z (%): 647.2311 [M\u2212SrI2+Na]+ (78), 839.0525 [M\u2212I]+ (35); IR: 2946 (w), 2885 (w), 1595 (w), 1500 (s), 1456 (m), 1400 (w), 1366 (w), 1250 (vs.), 1191 (m), 1161 (m), 1115 (m), 1063 (s), 934 (vs.), 916 (vs.), 862 (m), 837 (m), 817 (s), 774 (s), 750 (vs.), 734 (s), 711 (s), 630 (w), 605 (w), 533 (w), 516 (w), 484 (w), 456 (w), 422\u2005cm\u22121 (w). CHN calcd for C28H48I2O8Si4Sr\u22c50.5\u2009CH2Cl2: C, 33.93; H, 4.90. Found: C, 33.92; H, 4.68.6 was dissolved in DCM (10\u2005mL) and [222]cryptand was added. The resulting suspension was stirred overnight to give a clear solution. The solvent was removed under reduced pressure to give an oily, greasy residue. Extracting with n\u2010pentane (10\u2005mL) and subsequent filtering of the suspension yielded a clear solution. Removing the solvent under reduced pressure gave a greasy crown ether\u2010[222]cryptand mixture (0.13\u2005g). [222]crypt contamination was more than 30\u2009%.Compound 1H\u2005NMR : \u03b4= 0.24 , 3.96\u20134.01 , 4.02\u20134.08 , 6.91\u2005ppm ; 13C{1H}\u2005NMR : \u03b4= 0.8 , 63.5 , 72.5 , 116.7 , 122.5 , 150.5\u2005ppm ; 29Si{1H}\u2005NMR : \u03b4= 11.5\u2005ppm . MS: HR\u2010ESI(+) m/z (%): 625.2502 [M+H]+ (100), 647.2320 [M+Na]+ (100). IR: 2945 (w), 2869 (w), 1593 (w), 1501 (s), 1452 (m), 1396 (w), 1370 (w), 1328 (w), 1247 (s), 1221 (m), 1124 (s), 1098 (s), 1053 (s), 948 (m), 923 (m), 823 (m), 790 (s), 763 (vs.), 740 (s), 631 (m), 450\u2005cm\u22121 (w).3\u2009a, 5), STOE IPDS2 , STOE IPDS2T (2\u2009a) or a STOE STADIVARI (4\u2009a) diffractometer, respectively. Measurements were performed at 100\u2005K with MoK\u03b1 (\u03bb=0.71073\u2005\u00c5) or CuK\u03b1 (\u03bb=1.54184\u2005\u00c5) radiation, graphite monochromatization or respective X\u2010ray optics. The structures were solved by direct methods and refinement with full\u2010matrix\u2010least\u2010squares against F2 using SHELXT and SHELXL on OLEX2 platform.3\u2009a and 6, we applied the SQUEEZE operation. A selection of respective crystal data is given below.Single\u2010crystal X\u2010ray diffraction experiments were carried out on a Bruker D8 Quest \u2005K, a=8.173(3), b=12.204(4), c=14.204(8)\u2005\u00c5; \u03b2=93.02(4)\u00b0, V=1414.8(11)\u2005\u00c53, \u03c1=1.680\u2005g\u2009cm\u22123, numerical absorption correction using STOE X\u2010AREA and X\u2010RED32.\u03bc=2.618\u2005mm\u22121, Tmin, Tmax=0.5303, 0.8053, 2\u0398 range 4.402\u201358.996\u00b0, reflections measured 21\u2009645, independent reflections 7891 0.0244, wR2 0.0590, GoF 1.047, \u0394\u03c1max, \u0394\u03c1min 1.27/\u22120.52\u2005e\u2009\u00c53.Crystal data of 2\u2009a: C14H36CaI6O5Si4, monoclinic, C2/c, Z=4, 100(2)\u2005K, a=11.408(3), b=12.970(2), c=25.570\u2005\u00c5; \u03b2=92.38(2)\u00b0, V=3484.7(16)\u2005\u00c53, \u03c1=2.284\u2005g\u2009cm\u22123, spherical absorption correction using STOE X\u2010AREA and LANA.\u03bc=5.654\u2005mm\u22121, Tmin, Tmax=0.1989, 0.4381, 2\u0398 range 3.458\u201350.994\u00b0, reflections measured 13\u2009999, independent reflections 3250 [R(int)=0.0924], 154 parameters, R\u2010index [I\u22652\u03c3(I)] 0.0548, wR2 0.1400, GoF 1.072, \u0394\u03c1max, \u0394\u03c1min 1.56/\u22120.91\u2005e\u2009\u00c53.Crystal data of 3\u2009a: C16H44Ga2I8O5Si6Sr, monoclinic, P21/c, Z=4, 100(2)\u2005K, a=23.3580(16), b=11.3585(7), c=20.6920(14)\u2005\u00c5; \u03b2=111.998(4)\u00b0, V=5090.2(6)\u2005\u00c53, \u03c1=2.254\u2005g\u2009cm\u22123, spherical absorption correction using STOE X\u2010AREA and LANA,\u03bc=7.110\u2005mm\u22121, Tmin, Tmax=0.4793, 0.7452, 2\u0398 range 4.232\u201350.738\u00b0, reflections measured 171\u2009072, independent reflections 9287 [R(int)=0.1514], 355 parameters, R\u2010index [I\u22652\u03c3(I)] 0.0675, wR2 0.1863, GoF 1.076, \u0394\u03c1max, \u0394\u03c1min 4.17/\u22121.57\u2005e\u2009\u00c53.Crystal data of 4\u2009a: C16H40I4O6Si4Sr, monoclinic, P21/c, Z=4, 100(2)\u2005K, a=12.0422(5), b=22.2125(6), c=12.8413(4)\u2005\u00c5; \u03b2=95.734(3)\u00b0, V=3417.7(2)\u2005\u00c53, \u03c1=2.014\u2005g\u2009cm\u22123, spherical absorption correction using STOE X\u2010AREA and LANA,\u03bc=32.123\u2005mm\u22121, Tmin, Tmax=0.212, 0.360, 2\u0398 range 7.378\u201350.994\u00b0, reflections measured 26\u2009969, independent reflections 6337 [R(int)=0.0854], 288 parameters, R\u2010index [I\u22652\u03c3(I)] 0.0391, wR2 0.0894, GoF 0.804, \u0394\u03c1max, \u0394\u03c1min 1.23/\u22121.51\u2005e\u2009\u00c53.Crystal data of 5: C20H48I2O8Si4Sr, triclinic, P\u20101, Z=2, 110(2)\u2005K, a=8.4870(4), b=14.9819(7), c=15.4929(7)\u2005\u00c5, \u03b1=105.457(2)\u00b0, \u03b2=102.187(2)\u00b0, \u03b3=99.376(2)\u00b0, V=1804.98(15)\u2005\u00c53, \u03c1=1.601\u2005g\u2009cm\u22123, multiscan absorption correction using SADABS2016[50],\u03bc=3.371\u2005mm\u22121, Tmin, Tmax=0.5094, 0.8894, 2\u0398 range 4.642\u201357.786\u00b0, reflections measured 54\u2009208, independent reflections 9456 [R(int)=0.0442], 365 parameters, R\u2010index [I\u22652\u03c3(I)] 0.0323, wR2 0.0540, GoF 1.034, \u0394\u03c1max, \u0394\u03c1min 1.21/\u22120.80\u2005e\u2009\u00c53.Crystal data of 6\u22c51.5DCM: C29.5H51Cl3I2O8Si4Sr, triclinic, P\u20101, Z=4, 100(2)\u2005K, a=11.9953(8)\u2005\u00c5, b=16.0514(12)\u2005\u00c5, c=24.936(2)\u2005\u00c5, \u03b1=76.567(6)\u00b0, \u03b2=85.494(6)\u00b0, \u03b3=86.477(6)\u00b0, V=4650.7(6)\u2005\u00c53, \u03c1=1.562\u2005g\u2009cm\u22123, numerical absorption correction using STOE X\u2010AREA and X\u2010RED32,\u03bc=2.801\u2005mm\u22121, Tmin, Tmax=0.5094, 0.8894, 2\u0398 range 4.642\u201357.786\u00b0, reflections measured 52\u2009818, refined as a two component twin, 901 parameters, R\u2010index [I\u22652\u03c3(I)] 0.0780, wR2 0.2325, GoF 0.871, \u0394\u03c1max, \u0394\u03c1min 2.32/\u22121.74\u2005e\u2009\u00c53.The authors declare no conflict of interest."} +{"text": "The mol\u00adecule has an E configuration about the C=C bond and the carbonyl group is syn with respect to the C=C bond. In the crystal, the mol\u00adecules are linked along the a-axis direction through van der Waals forces and by face-to-face \u03c0-stacking between the thio\u00adphene rings and between the benzene rings of neighbouring mol\u00adecules along the b axis into zigzag sheets lying parallel to the bc plane.The mol\u00adecular structure of the title compound consists of a 2,5-di\u00adchloro\u00adthio\u00adphene ring and a 2-chloro\u00adphenyl ring linked 13H7Cl3OS, consists of a 2,5- di\u00adchloro\u00adthio\u00adphene ring and a 2-chloro\u00adphenyl ring linked via a prop-2-en-1-one spacer. The dihedral angle between the 2,5-di\u00adchloro\u00adthio\u00adphene and 2-chloro\u00adphenyl rings is 9.69\u2005(12)\u00b0. The mol\u00adecule has an E configuration about the C=C bond and the carbonyl group is syn with respect to the C=C bond. The mol\u00adecular conformation is stabilized by two intra\u00admolecular C\u2014H\u22efCl contacts and one intra\u00admolecular C\u2014H\u22efO contact, forming S(5)S(5)S(6) ring motifs. In the crystal, the mol\u00adecules are linked along the a-axis direction through van der Waals forces and along the b axis by face-to-face \u03c0-stacking between the thio\u00adphene rings and between the benzene rings of neighbouring mol\u00adecules, forming corrugated sheets lying parallel to the bc plane. The inter\u00admolecular inter\u00adactions in the crystal packing were further analysed using Hirshfield surface analysis, which indicates that the most significant contacts are Cl\u22efH/ H\u22efCl (28.6%), followed by C\u22efH/H\u22efC (11.9%), C\u22efC (11.1%), H\u22efH (11.0%), Cl\u22efCl (8.1%), O\u22efH/H\u22efO (8.0%) and S\u22efH/H\u22efS (6.6%).The mol\u00adecular structure of the title compound, C The mol\u00adecules are packed into corrugated sheets lying parallel to (011) Figs. 2 and 3 \u25b8.CrystalExplorer ] with de + di \u2243 2.85\u2005\u00c5 . The C\u22efH/H\u22efC inter\u00adactions are shown in Fig.\u00a04c). The scattered points show the van der Waals contacts and \u03c0\u2013\u03c0 stacking inter\u00adactions. The inter\u00adatomic C\u22efC contacts appear as an arrow-shaped distribution of points in Fig.\u00a04d), with the vertex at de = di = 1.75\u2005\u00c5. The C\u22efC contacts reflect \u03c0\u2013\u03c0 inter\u00adactions between the aromatic rings. The H\u22efH inter\u00adactions are reflected in Fig.\u00a04e) as widely scattered points of high density due to the large hydrogen content of the mol\u00adecule. The split spike with the tip at de = di \u2243 1.3\u2005\u00c5 is due to the short inter\u00adatomic H\u22efH contacts. Cl\u22efCl contacts [Fig.\u00a04f)] are disfavoured when the number of H atoms on the mol\u00adecular surface is large because of competition with the more attractive H\u22efCl contacts. Cl\u22efCl contacts from a parallel alignment of C\u2014Cl bonds may be indicated. They are known in the literature as type-I halogen\u2013halogen inter\u00adactions ] also have a symmetrical distribution of points, with two pairs of thin and thick edges at de + di \u2243 2.75\u2005\u00c5. The S\u22efH contacts shown in Fig.\u00a04h) are contracted to a much lesser degree.The C\u2014H\u22efCl inter\u00adactions appear as two distinct spikes in the fingerprint plot -3-(thio\u00adphen-2-yl)prop-2-en-1- one [(X); Yesilyurt et al., 2018E)-1-(2-amino\u00adphen\u00adyl)-3- (thio\u00adphen-2-yl)prop-2-en-1-one [(XI); Chantrapromma et al., 2013et al., 2018P1, while (III) and (IV) are isostructural in space group P21/c. There are no hydrogen bonds of any kind in the structures of compounds (I)C(7) chains by means of C\u2014H\u22efO hydrogen bonds. In (V), there are again no hydrogen bonds nor any \u03c0\u2013\u03c0 stacking inter\u00adactions but in (VI), the mol\u00adecules are linked into C(5) chains by C\u2014H\u22efO hydrogen bonds. In each of compounds (I)\u2013(VI), the mol\u00adecular skeletons are close to planarity, and there are short halogen\u2013halogen contacts in the structures of compounds (II) and (V) and a short Br\u22efO contact in the structure of compound (VI). In (VII), the mol\u00adecule is non-planar, with a dihedral angle of 22.6\u2005(2)\u00b0 between the aromatic rings. The mol\u00adecules are linked by pairs of C\u2014H\u22ef\u03c0 inter\u00adactions, forming inversion dimers. There are no other significant inter\u00admolecular inter\u00adactions present. In (VIII), the mol\u00adecule is nearly planar, the dihedral angle between the thio\u00adphene and phenyl rings being 9.07\u2005(8)\u00b0. The mol\u00adecules are linked via weak C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds, forming chains propagating along the c-axis direction. In (IX), the thienyl ring is not coplanar with the benzene ring, their planes forming a dihedral angle of 13.2\u2005(4)\u00b0. In the crystal, mol\u00adecules stack along the a-axis direction, with the inter\u00adplanar separation between the thienyl rings and between the benzene rings being 3.925\u2005(6)\u2005\u00c5. In (X), the thio\u00adphene ring forms a dihedral angle of 26.04\u2005(9)\u00b0 with the benzene ring. The mol\u00adecular conformation is stabilized by an O\u2014H\u22efN hydrogen bond. The mol\u00adecules are connected through C\u2014H\u22efO hydrogen bonds, forming wave-like layers parallel to the ab plane, which are further linked into a three-dimensional network by C\u2014H\u22ef\u03c0 inter\u00adactions. In (XI), the mol\u00adecule is almost planar with a dihedral angle of 3.73\u2005(8)\u00b0 between the phenyl and thio\u00adphene rings. An intra\u00admolecular N\u2014H\u22efO hydrogen bond generates an S(6) ring motif. Adjacent mol\u00adecules are linked into dimers in an anti-parallel face-to-face manner by pairs of C\u2014H\u22efO inter\u00adactions. Neighbouring dimers are further linked into chains along the c-axis direction by N\u2014H\u22efN hydrogen bonds. In (XII), the dihedral angle between the thio\u00adphene and benzene rings increases to12.24\u2005(15)\u00b0. The mol\u00adecular conformation is stabilized by intra\u00admolecular C\u2014H\u22efCl contacts, forming S(6) and S(5) ring motifs. In the crystal, the mol\u00adecules are linked through face-to-face \u03c0-stacking between the thio\u00adphene rings and the benzene rings of the mol\u00adecules into zigzag sheets lying parallel to the bc plane.The closest related compounds with the same skeleton and containing a similar bis-chalcone moiety to the title compound but with different substituents on the aromatic rings are: = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989018018066/qm2131sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018018066/qm2131Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018018066/qm2131Isup3.cmlSupporting information file. DOI: 1036796CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the adduct contains a neodymium(III) cation, three coordinated nitrate anions, four coordinated water mol\u00adecules and three uncoordinated neutraltriazine mol\u00adecules. The crystal structure consists of a three-dimensional supra\u00admolecular framework held together by a network of O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds between the coordinated water mol\u00adecules, nitrate ions and triazine mol\u00adecules. The uncoordinated neutral triazine moieties form N\u2014H\u22efN hydrogen bonds. Hirshfeld surface and fingerprint plots identify the major contributors to the inter\u00admolecular inter\u00adactions. 3)3(H2O)4]\u00b73C3H4N4, neodymium is ten-coordinate with a distorted bicapped square-anti\u00adprismatic geometry formed from six O atoms from three nitrate ions and four O atoms from four coordinated water mol\u00adecules. The structure also contains neutral 3-amino-1,2,4-triazine mol\u00adecules which are not coordinated to the central metal atom. The coordinated water mol\u00adecules and nitrate ions of adjacent complexes are linked by O\u2014H\u22efO hydrogen bonds to form cyclic R22(8) ring motifs, which in turn are further connected via hydrogen bonds to generate a sheet-like structure. The triazine mol\u00adecules are involved in a number of hydrogen-bonding inter\u00adactions: N\u2014H\u22efN and O\u2014H\u22efN inter\u00adactions to form R33(9) motifs and N\u2014H\u22efN inter\u00adactions to link the organic mol\u00adecules into chains. Weak C\u2014H\u22efO hydrogen bonds also occur between triazine mol\u00adecules and coordinated nitrate atoms. All these inter\u00admolecular contacts contribute to the stabilization of the three-dimensional supra\u00admolecular framework. Hirshfeld surface analysis shows that N\u22efH/H\u22efN and H\u22efH inter\u00adactions account for 42.9 and 20.6% of the surface, respectively.In the title compound, [Nd(NO All these inter\u00admolecular inter\u00adactions appear to play a significant role in stabilizing the crystal structure and result in the formation of a three-dimensional supra\u00admolecular framework is based on the distances from the nearest atom inside (di) and outside (de) the surface. The three-dimensional dnorm surface of the title compound is shown in Fig.\u00a05dnorm values on the surface correspond to the N\u2014H\u22efN, N\u2014H\u22efO and O\u2014H\u22efO inter\u00adactions. Analysis of the two-dimensional fingerprint plots reveal that the H\u22efH (20.6%) and N\u22efH/H\u22efN (42.9%) inter\u00adactions are the highest contributors to the Hirshfeld surface. Smaller contributions come from O\u22efH/H\u22efO (13.3%) C\u22efH/H\u22efC (6.3%), N\u22efN (6.2%), C\u22efN/N\u22efC (4.6%), N\u22efO/O\u22efN (2.8%) and C\u22efO/O\u22efC (1.8%) inter\u00adactions -biscopper(II)] and KUCNEC [with bis\u00adhexa\u00adkis\u00adtricopper(II)] \u00adbis\u00ad\u00adcobalt]; Palion-Gazda et al., 2015catena-[bis\u00ad(\u03bc2-dicyanamido)\u00adbis\u00ad\u00adcobalt]; \u015ewitlicka-Olszewska et al., 2016A search of the Cambridge Structural Database 3\u00b76H2O (0.219g) (Alfa Aesar). Di\u00adchloro\u00admethane (5\u2005ml) was then added and the mixture refluxed for 7\u2005h at 353\u2005K. The resulting solution was then allowed to cool slowly to room temperature. After two weeks, brown-coloured crystals were obtained, m.p. = 378\u2005K.The title compound was prepared by adding a hot methano\u00adlic solution (20\u2005ml) of 3-amino-1,2,4-triazine (0.043g) (Aldrich) to a hot methano\u00adlic solution (20\u2005ml) of Nd(NOUiso(H) set to 1.2\u20131.5Ueq(C). The water and N-bound H atoms were located in difference-Fourier maps and refined with Uiso(H) = 1.2Ueq(O) or 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018011714/cq2026sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018011714/cq2026Isup2.hklStructure factors: contains datablock(s) I. DOI: 1583097CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The conformations of the title compounds, (I) and (II), are very similar. The pyran rings adopt envelope conformations, the piperidine rings have chair conformations and the pyrrolidine rings adopt twist conformations. Intra- and inter\u00admolecular C\u2014H\u22efO hydrogen bonds occur. Compound (II) crystallizes with two independent mol\u00adecules in the asymmetric unit which are linked by C\u2014H\u22efO hydrogen bonds. 32H25ClN2O4 (I) and C33H28N2O5 (II), the six-membered pyran and piperidine rings adopt envelope and chair conformations, respectively. The five-membered pyrrolidine rings adopt twist conformations. Compound (II) crystallizes with two independent mol\u00adecules (A and B) in the asymmetric unit. In all three mol\u00adecules there is a C\u2014H\u22efO intra\u00admolecular hydrogen bond present enclosing an S(7) ring motif. In (I), both oxygen atoms of the nitro group are disordered, while in (II) the meth\u00adoxy\u00adbenzene group is disordered in mol\u00adecule B. The geometries were regularized by soft restraints. In the crystal of (I), mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, forming chains along [010]. The chains are linked by C\u2014H\u22efCl hydrogen bonds, forming layers parallel to (10A and B mol\u00adecules are linked via C\u2014H\u22efO hydrogen bonds, forming a square four-membered A\u2013B\u2013A\u2013B unit. These units are linked by a number of C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional supra\u00admolecular structure.In the title compounds, C The structural overlap of compound (I)B of compound (II)B of compound II (major component) on mol\u00adecule A is shown in Fig.\u00a05A and IIB) the pyran rings have envelope conformations with the methyl\u00adene C atom C21 as the flap. The piperidine rings adopt chair conformations, while the pyrrolidine rings adopt twist conformations on the N1\u2014C12 bond (N1A\u2014C12A in IIA and N1B\u2014C12B in IIB).The mol\u00adecular structure of compound (I)A and B of compound (II)A and 66.7\u2005(2) and 59.3\u2005(5)\u00b0 for mol\u00adecule B. The benzene rings (C27\u2013C32 and C14\u2014C19) are inclined to each other by 50.0\u2005(1)\u00b0 in (I)A and 71.6\u2005(2)\u00b0 in mol\u00adecule B of (II)A and 36.1\u2005(2)\u00b0 for mol\u00adecule B in (II)A and 13.1\u2005(2)\u00b0 in mol\u00adecule B of compound (II)A and 36.2\u2005(2)\u00b0 in mol\u00adecule B of compound (II)supporting information. The keto atom O1 deviates from the mean plane of the plane of the ace\u00adnaphthyl\u00adene ring system (C1\u2014C12) by 0.070\u2005(2)\u2005\u00c5 in (I)et al., 2013abThe mean plane of the five-membered pyrrolidine ring (N1/C12/C13/C21/C22) is inclined to the mean plane of the cyclo\u00adpen3\u2013en-1-one ring (C1/C2/C10\u2013C12) by 85.7\u2005(1)\u00b0 in compound (I)b-axis direction. The chains are linked by C\u2014H\u22efCl hydrogen bonds, forming layers parallel to the s Table\u00a02. These as Table\u00a02.et al., 2016H,6\u2032H,6b\u2032H-spiro\u00adindolizin]-2-one skeleton yielded two hits: namely 6-(4-meth\u00adoxy\u00adphen\u00adyl)-6a-nitro-6,6a,6b,7,8,9,10,12a-octa\u00adhydro\u00adspiro\u00adindolizine-12,3-indolin]-2-one for (I)H-chromene (1\u2005mmol) for (II)To a solution of ace\u00adnaphtho\u00adquinone (1.0\u2005mmol) and piperidine-2-carb\u00adoxy\u00adlic acid (1.5\u2005mmol) in dry toluene, was added 2-(4-chloro\u00adphen\u00adyl)-3-nitro-2Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. In compound (I)B is disordered, as detectable from the large displacement parameters for the C and O atoms and short C\u2014C and C\u2014O bond lengths. This disorder over two positions was modelled and the site occupancies refined to 0.739\u2005(5) and 0.261\u2005(5). The geometry was regularized by soft restraints.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019000422/su5472sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: Click here for additional data file.10.1107/S2056989019000422/su5472Isup2.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019000422/su5472IIsup3.cmlSupporting information file. DOI: 10.1107/S2056989019000422/su5472sup4.pdfPuckering and asymmety parameters. DOI: 1024915, 1024235CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Correction to: Infect Agents Cancer (2019) 14:46https://doi.org/10.1186/s13027-019-0262-5The original publication of this article containeErrors in original publication:\u201cFish notch1b/Human NOTCH2\u201d should be \u201cFish reck/Human RECK\u201cFish notch1b/Human NOTCH3\u201d should be \u201cFish srd5a1/Human SRD5A1\u201cFish notch1b/Human NOTCH4\u201d should be \u201cFish wnt7bb/ HumanWNT7B\u201d\u201cFish notch1b/Human NOTCH5\u201d should be \u201cFish pparg/Human PPARG\u201d"} +{"text": "Nature Communications 10.1038/s41467-019-12167-9, published online 19 September 2019.Correction to: S\u03bb(P)\u2009=\u20091.45, WP\u2009=\u20091.08 and W\u03c9\u2009=\u200932.3\u2019. The correct version reads \u2018S\u03bb(P)\u2009=\u20090.90, WP\u2009=\u20090.71 and W\u03c9\u2009=\u200925.3\u2019. Similarly in Fig.\u00a01b, the label at the top of the left panel incorrectly read \u2018S\u03bb(P)\u2009=\u20090.90, WP\u2009=\u20090.71 and W\u03c9\u2009=\u200925.3\u2019. The correct version reads \u2018S\u03bb(P)\u2009=\u20091.45, WP\u2009=\u20091.08 and W\u03c9\u2009=\u200932.3\u2019. This has been corrected in both the PDF and HTML versions of the Article.The original version of this Article contained errors in Fig.\u00a01. In Fig.\u00a01a, the label at the top of the left panel incorrectly read \u2018"} +{"text": "Correction to: Microbiomehttps://doi.org/10.1186/s40168-019-0643-4Following publication of the original article , the autPage 3, right column: Additional\u00a0file\u00a01: Table S1\u2009\u2192\u2009Table\u00a01.Page 4, right column: detail in Additional\u00a0file\u00a01: Table S1\u2009\u2192\u2009detail in Additional\u00a0file\u00a02: Supplementary results.Page 5, right column: Table\u00a02 \u2009\u2192\u2009Table\u00a02 .Page 12, left column: Additional\u00a0file\u00a01: Table S10\u2009\u2192\u2009Additional\u00a0file\u00a01: Table S11.Page 12, left column: Additional\u00a0file\u00a01: Table S12\u2009\u2192\u2009Additional\u00a0file\u00a05: Table S12.Description of additional files on Page 12 and 13:Table S10 and Table\u2009S12\u2192\u2009Table S12 and Table S10. Respectively.Additional file 5 Table S5\u2009\u2192\u2009Additional file 5: Table S12.Kim et al. [19]\u2009\u2192\u2009Kim et al. [21]"} +{"text": "In the crystal, the mol\u00adecules form zigzag stacks along the 15H12ClNO3, consists of a 1,2-di\u00adhydro\u00adquinoline-4-carb\u00adoxyl\u00adate unit with 2-chloro\u00adethyl and propynyl substituents, where the quinoline moiety is almost planar and the propynyl substituent is nearly perpendicular to its mean plane. In the crystal, the mol\u00adecules form zigzag stacks along the a-axis direction through slightly offset \u03c0-stacking inter\u00adactions between inversion-related quinoline moieties which are tied together by inter\u00admolecular C\u2014HPrpn\u00adyl\u22efOCarbx and C\u2014HChlethy\u22efOCarbx hydrogen bonds. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (29.9%), H\u22efO/O\u22efH (21.4%), H\u22efC/C\u22ef H (19.4%), H\u22efCl/Cl\u22efH (16.3%) and C\u22efC (8.6%) inter\u00adactions. Hydrogen bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Computational chemistry indicates that in the crystal, the C\u2014HPrpn\u00adyl\u22efOCarbx and C\u2014HChlethy\u22efOCarbx hydrogen bond energies are 67.1 and 61.7\u2005kJ\u2005mol\u22121, respectively. Density functional theory (DFT) optimized structures at the B3LYP/ 6\u2013311\u2005G level are compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.The title compound, C We report herein on the synthesis and the mol\u00adecular and crystal structures of the title compound along with the Hirshfeld surface analysis and the inter\u00admolecular inter\u00adaction energies and the density functional theory (DFT) computational calculation carried out at the B3LYP/6\u2013311\u2005G level.The quinoline ring system is an important structural unit in naturally occurring quinoline alkaloids, therapeutics and synthetic analogues with inter\u00adesting biological activities. Quinolone derivatives possess a variety of pharmacological properties such as anti-bacterial (Hu A (C1\u2013C6) and B (N1/C1/C6\u2013C9), of the di\u00adhydro\u00adquinoline unit are oriented at a dihedral angle of 2.69\u2005(17)\u00b0. The mean plane through the di\u00adhydro\u00adquinoline unit is almost planar with a maximum deviation of 0.040\u2005(3)\u2005\u00c5 for atom N1, and the propynyl substituent is nearly perpendicular to that plane, the C6\u2014N1\u2014C10\u2014C11 torsion angle being \u221279.6\u2005(4)\u00b0. The carboxyl group is twisted out of coplanarity with the di\u00adhydro\u00adquinoline unit by a dihedral angle of 47.13\u2005(23)\u00b0; this is also indicated by the C1\u2014C9\u2014C13\u2014O2 torsion angle of \u221244.2\u2005(6)\u00b0.The title mol\u00adecule consists of a 1,2-di\u00adhydro\u00adquinoline-4-carboxyl\u00adate unit with 2-chloro\u00adethyl and propynyl substituents Fig.\u00a01. The cona-axis direction through slightly offset \u03c0-stacking inter\u00adactions between inversion-related quinoline moieties hydrogen bonds, enclosing A (C1\u2013C6) and B (N1/C1/C6\u2013C9), of the di\u00adhydro\u00adquinoline unit, Cg2\u22efCg1i, Cg2\u22efCg1ii and Cg1\u22efCg1i , may further stabilize the structure.In the crystal, the mol\u00adecules form zigzag stacks along the es Fig.\u00a02. The stas Table\u00a01. The \u03c0\u2013\u03c0CrystalExplorer17.5 analysis are viewed as a pair of spikes with the tips at de + di = 2.28\u2005\u00c5. In the absence of C\u2014H\u22ef\u03c0 inter\u00adactions, the pairs of characteristic wings in Fig.\u00a07d arise from H\u22efC/C\u22efH contacts (19.4%) and are viewed as pairs of spikes with the tips at de + di = 2.65\u2005\u00c5 and 2.70\u2005\u00c5 for the thin and thick spikes, respectively. The scattered points in the pair of wings in the fingerprint plot delineated into H\u22efCl/Cl\u22efH have a symmetrical distribution with the edges at de + di = 2.60\u2005\u00c5. The C\u22efC contacts, Fig.\u00a07f, have an arrow-shaped distribution of points with the tip at de = di = 1.72\u2005\u00c5. Finally, the characteristic tip and wings in the fingerprint plots delineated into C\u22efN/N\u22efC and O\u22efCl/Cl\u22efO contacts have the tips at de = di = 1.73 and 3.70\u2005\u00c5, respectively.The overall two-dimensional fingerprint plot, Fig.\u00a07H Table\u00a02, contribdnorm plotted onto the surface are shown for the H\u22efH, H\u22efO/O\u22efH, H\u22efC/C\u22efH and H \u22ef Cl/Cl\u22efH inter\u00adactions in Fig.\u00a08a\u2013d, respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efO/O\u22efH, H \u22ef C/C\u22efH and H\u22efCl/Cl\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing is the sum of electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies were calculated to be \u221225.2 (Eele), \u22122.1 (Epol), \u221285.4 (Edis), 57.5 (Erep) and \u221267.1 (Etot) for the C\u2014HPrpn\u00adyl\u22efOCarbx hydrogen bond and \u221226.5 (Eele), \u22124.7 (Epol), \u221273.2 (Edis), 54.3 (Erep) and \u221261.7 (Etot) for the C\u2014HChlethy\u22efOCarbx hydrogen bond.The inter\u00admolecular inter\u00adaction energies were calculated using the CE\u2013B3LYP/6\u201331G energy model available in via density functional theory (DFT) using the standard B3LYP functional and 6\u2013311\u2005G basis-set calculations . The significance of \u03b7 and \u03c3 is to evaluate both the reactivity and stability. The electron transition from the HOMO to the LUMO energy level is shown in Fig.\u00a09E = ELUMO\u00a0\u2212\u00a0EHOMO] of the mol\u00adecule is 3.6984\u2005eV, and the frontier mol\u00adecular orbital energies, EHOMO and ELUMO are \u22126.3024 and \u22122.6040\u2005eV, respectively.The optimized structure of the title compound in the gas phase was generated theoretically et al., 2016et al., 2018abet al., 2019A non-alkyl\u00adated analogue, namely quinoline and its derivatives, has been reported and TBAB . The reaction mixture was stirred at room temperature for 6\u2005h. After removal of the salts by filtration, the solvent was evaporated under reduced pressure and the resulting residue was dissolved in di\u00adchloro\u00admethane. The organic phase was dried with Na2SO4, and then concentrated under reduced pressure. The pure compound was obtained by column chromatography using hexa\u00adne/ethyl acetate (3/1) as eluent. The isolated solid was recrystallized from hexa\u00adne/ethyl acetate (3:1) to afford colourless crystals .To a solution of 2-chloro\u00adethyl 2-oxo-1,2-di\u00adhydro\u00adquinoline-4-carboxyl\u00adate in DMF (10.00\u2005ml) were added propargyl bromide , K2 H atoms, respectively) and constrained to ride on their parent atoms, with Uiso(H) = 1.2Ueq(C). The largest peak and hole in the final difference map are +0.73\u2005e\u2005\u00c5\u22123 (1.00\u2005\u00c5 away from Cl1) and \u22120.35\u2005e\u2005\u00c5\u22123 (0.64\u2005\u00c5 away from C14), and are associated with the 2-chloro\u00adethyl\u00adcarb\u00adoxy group and may indicate a slight degree of disorder here but it was not considered serious enough to model.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989019012283/lh5918sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019012283/lh5918Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019012283/lh5918Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019012283/lh5918Isup4.cmlSupporting information file. DOI: 1951439CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Quantum chemical calculations revealed details on the strength and geometrical requirements of these N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds, and a subsequent survey of the Protein Data Bank (PDB) based on these criteria suggested that numerous protein-ligand complexes contain such N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds. An ensuing experimental investigation into the G9a-like protein (GLP)-inhibitor complex demonstrated that N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds affect the activity of the inhibitors against the target enzyme. These results should provide the basis for the use of N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds in drug discovery.In the context of drug design, C-H\u00b7\u00b7\u00b7O hydrogen bonds have received little attention so far, mostly because they are considered weak relative to other noncovalent interactions such as O-H\u00b7\u00b7\u00b7O hydrogen bonds, \u03c0/\u03c0 interactions, and van der Waals interactions. Herein, we demonstrate the significance of hydrogen bonds between C-H groups adjacent to an ammonium cation and an oxygen atom (N Such interactions manifest between proteins and their ligands in many protein complexes registered on the Protein Data Bank (PDB). Accordingly, these interactions should always be considered when designing ligands for target proteins7.Noncovalent interactions, such as heteroatom-hydrogen bonds X-H\u00b7\u00b7\u00b7Y and \u03c0/\u03c0 interactions play a critical role in the formation of protein-ligand complexes11. However, when the C-H group is activated by electron-withdrawing groups, e.g. C-H groups that are covalently bound to a cationic nitrogen atom (N+-C-H), C-H\u00b7\u00b7\u00b7O hydrogen bonds may become as strong as heteroatom-hydrogen bonds, which could be important for molecular recognition19. For example, N+-C-H\u00b7\u00b7\u00b7Y hydrogen bonds are likely to be involved in the substrate recognition of tetraalkylammonium-based catalysts21, and the ligand/substrate recognition in receptors/enzymes may be controlled by N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds , the latter are almost exhaustively protonated under physiological conditions (pH\u2009=\u20097\u20138). Accordingly, ligands bearing tetraalkylammonium or aliphatic amino groups could be recognized by the oxygen atoms of the proteins via N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds theoretical research and criteria setting for N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds by quantum chemical calculations , propanoate (2), ethanol (3), and phenol (4) as hydrogen acceptor models for the protein peptide bond/Asn/Gln, Asp/Glu, Ser/Thr, and Tyr, respectively, and monomethylammonium (5), trimethylammonium (6) or N-methylpiperidium (7) as hydrogen donor models in protonated aliphatic amine-containing ligands. Geometry optimizations and single-point calculations were carried out at the M06-2X/6-311++G** level of theory26. The results of this computational study allowed us to establish selection criteria for N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds analysis28 of 5 indicated that the positive charge is distributed over the H atoms , including those of the CH3 group, rather than being localized on the N atom (\u22120.688) is more positive than that of monomethylamine or ethane . These results suggest high potential for the N+-C-H group to act as a hydrogen donor in hydrogen bonds.Initially, we analyzed the electrical charges on the hydrogen donors of the NA1, B1 and C1, i.e., complexes of 1, 2, and 3 with 6, as well as that of complex D1, which consists of 4 and 7 in the gas phase , which suggests the formation of intermolecular hydrogen bonds between these atoms or 2,2-dimethylpropane (11), and 7 in D1 is replaced with N-methylpiperidine (12) or methyl cyclohexane (13) , the C-H\u00b7\u00b7\u00b7O angle (\u03c8), the H\u00b7\u00b7\u00b7O=C/H\u00b7\u00b7\u00b7O-C angle (\u03be), and the H-elevation angle (\u03b8)30 were calculated 30 Figs\u00a0 and 5A. ant Fig.\u00a0. For H\u00b7\u00b7ted Fig.\u00a0, which ited Fig.\u00a0. The est7\u2009\u00c5 Fig.\u00a0. The dis\u2009\u00c5) Figs\u00a0 and 5B. A4 on the C-H\u00b7\u00b7\u00b7O angle (\u03c8). For 90\u00b0\u2009<\u2009\u03c8\u2009<\u2009180\u00b0, the interaction energy is relatively low Figs\u00a0 and 5C. ses Fig.\u00a0, a simil80\u00b0 Figs\u00a0 and 5C. \u03be). Calculating the interaction energies, we discovered that these energies depend on the type of hydrogen acceptors. In the case of amide acceptors, i.e., for 105\u00b0\u2009<\u2009\u03be\u2009<\u2009180\u00b0, the interaction energies are low and the preferred angles are \u03be\u2009>\u2009135\u00b0 was permissible, and B4 exhibited a preference for \u03be\u2009>\u2009240\u00b0 Figs\u00a0. For car\u03b8). For \u03b8\u2009<\u200970\u00b0 (A4), \u03b8\u2009<\u200960\u00b0 (C4), and \u03b8\u2009<\u200975\u00b0 (D4), low interaction energies were calculated and propanoate (2) are widely distributed and phenol (4) is centered on the area where the plane bisecting the C-O-H angle intersects with the C-O-H plane and H\u00b7\u00b7\u00b7O (dHO) bond distances in the \u03be dataset, we observed 13 interactions with 105\u00b0\u2009<\u2009\u03be\u2009<\u2009135\u00b0. According to our calculations and C-elevation (\u03d5) angles are subject to a certain degree of correlation , whereas contacts of carboxylate acceptors exhibited a wide distribution of these angles -inhibitor complex 14a . Relative to 14a, 14b significantly reduced the GLP-inhibitory activity (IC50\u2009=\u20090.664\u2009\u03bcM) in the 0.1\u20131\u2009\u03bcM concentration range. In the same concentration range, 14c showed a decreased GLP-inhibitory activity (IC50\u2009=\u20091.45\u2009\u03bcM) compared to 14a and 14b, while 14d did not show any GLP inhibition up to 3\u2009\u03bcM. The latter result should most likely be rationalized in terms of a lack in both electrostatic interactions and C-H\u00b7\u00b7\u00b7O hydrogen bonds between the ligand and GLP. Moreover, we determined the dissociation constant and thermodynamic parameters of 14a\u2013c by means of isothermal titration calorimetry (ITC). As shown in Supplementary Fig.\u00a014a\u2013c was distinctly dependent on the number of methyl group , which is consistent with the IC50 values of 14a\u2013c. The thermodynamic parameters were also dependent on the number of methyl group: the \u0394H values for dimethyl 14a, monomethyl 14b, and non-methyl 14c were \u221210.7\u2009kcal/mol, \u22128.58\u2009kcal/mol, and \u22127.04\u2009kcal/mol, respectively; the \u2212T\u0394S values for 14a, 14b, and 14c were 1.42\u2009kcal/mol, 0.10\u2009kcal/mol, and \u22121.19\u2009kcal/mol, respectively . It is well known that enthalpic forces such as hydrogen bond formation can decrease entropic forces by restricting the degrees of freedom of water molecules and protein conformation33. Based on this, there is a possibility that the methyl group of 14a and 14b reduced the \u0394H value by the conformational fixation of GLP through the formation of N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds, which resulted in reduction of the T\u0394S value by decreasing the degrees of freedom of water molecules and protein conformation, and the effect of enthalpy was larger than that of entropy. Taken together, the observed GLP-inhibitory activity, disassociation constant and thermodynamic parameters for 14a\u2013d strongly suggests that the N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds in the GLP-ligand complex are responsible for their GLP-inhibitory activity, although the effect might not be attributed exclusively to the N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds.To experimentally validate the importance of the N14a Fig.\u00a0, one of GLP Fig.\u00a0. In ordetom Fig.\u00a0. Compounely Fig.\u00a0. If the ity Fig.\u00a0. The par+-CH3 groups do not engage in N+-C-H\u00b7\u00b7\u00b7O hydrogen bonds with any protein-based oxygen atom. The X-ray crystal structure of tyrosine kinase with Ig and EGF homology domains-2 (tie-2) complexed to its inhibitor 15a indicates that the methylene group that is in conjugation with the aliphatic amino group forms an N+-C-H\u00b7\u00b7\u00b7O hydrogen bond, but the methyl groups do not 36, which is often used for long distance interactions, with the 6-311++G** basis set in the gas phase or in the water phase37. Structures were considered as minima in case of all harmonic frequencies being positive. All interaction energies were corrected for basis set superposition errors (BSSE) by the counterpoise procedure38. To gauge the accuracy of the M06-2X/6-311++G** level of theory, the thus obtained binding energies were compared to those obtained from MP2/aug-cc-pVTZ C\u00b7\u00b7\u00b7O distance (dCO): <3.7\u2009\u00c527; (ii) H\u00b7\u00b7\u00b7O distance (dHO): <2.7 \u00c5; (iii) C-H\u00b7\u00b7\u00b7O angle (\u03c8): >90\u00b0 for the peptide bond/Asn/Gln and Asp/Glu acceptors, or >105\u00b0 for the Ser/Thr/Tyr acceptors; (iv) H\u00b7\u00b7\u00b7O=C angle (\u03be): >105\u00b0 for the peptide bond/Asn/Gln acceptors, >90\u00b0 for the Asp/Glu acceptors, or 105\u00b0\u2013150\u00b0 for the Ser/Thr/Tyr acceptors; (v) H\u00b7\u00b7\u00b7elevation angle (\u03b8): <70\u00b0 for the peptide bond/Asn/Gln acceptors, <80\u00b0 for the Asp/Glu acceptors; (vi) N+-C\u00b7\u00b7\u00b7O angle (\u03b1): 60\u00b0\u2013160\u00b0; (vii) C\u00b7\u00b7\u00b7O-C angle (\u03b2): >95\u00b0 for the peptide bond/Asn/Gln, >90\u00b0 for the Asp/Glu acceptors, or 95\u00b0\u2013160\u00b0 for the Ser/Thr/Tyr acceptors; (viii) C-elevation (\u03d5): <80\u00b0 for the peptide bond/Asn/Gln and <90\u00b0 for Asp/Glu. The H-elevation angle (\u03b8) was calculated according to sin\u03b8\u2009=\u2009sin\u03be\u2009\u00b7\u2009sin\u03c7, whereby \u03c7 refers to the average of the two corresponding dihedral angles H\u00b7\u00b7\u00b7O=C-O(N) and H\u00b7\u00b7\u00b7O=C-C. Similarly, the C-elevation angle (\u03d5) was calculated by sin\u03d5\u2009=\u2009sin\u03b2\u2009\u00b7\u2009sin\u03b4, whereby \u03b4 refers to the average of the two corresponding dihedral angles C\u00b7\u00b7\u00b7O=C-O(N) and C\u00b7\u00b7\u00b7O=C-C.All X-ray structures of proteins bound to amine-containing ligands were downloaded from the Protein Data Bank (PDB). To gain a better understanding of NThe GLP activity assay was carried out using a GLP Chemiluminescent Assay Kit . Microwells were rehydrated by adding 200\u2009\u03bcL of tris buffered saline with Tween 20 to every well, followed by incubation at room temperature for 45\u2009minutes. After removing TBS-T, the inhibitors were incubated in the presence of 2\u2009\u03bcM SAM and 40\u2009ng of GLP in the supplied buffer on the microwells of . After the enzymatic reaction, every well was washed three times with TBS-T (100\u2009\u03bcL) and blocked for 10\u2009min with blocking buffer. Then, 100\u2009\u03bcL of primary anti-body solution (1:400 dilution) were added to the microwells, followed by incubation (2\u2009h). The wells were probed with the primary antibody, washed three times with TBS-T (100\u2009\u03bcL), incubated with sheep secondary anti-body (1:1000 diluted), and again washed three times with TBS-T (100\u2009\u03bcL). The chemiluminescence of the wells, to which detection reagents were added, was measured on a chemiluminescence reader (ARVO X3 Multilabel Plate Reader), and the values of % inhibition were calculated from the chemiluminescence readings of inhibited wells relative to those of control wells.\u22121. The heat of dilution of the ligand into the buffer measured independently was essentially similar to the heat signal obtained at the end of the titration; so the signal of the last injection was used as the background heat signal. The experimental data were analyzed using the MicroCal PEAQ-ITC Analysis software , and fitted using the one-site binding model. Values of \u0394G, KD, and \u2212T\u0394S were calculated using the thermodynamic relationships, \u0394G\u2009=\u2009\u2212RT lnKa, KD\u2009=\u20091/Ka, and \u0394G\u2009=\u2009\u0394H\u2009\u2212\u2009T\u0394S.A GLP recombinant protein solution was concentrated by ultrafiltration and the buffer was replaced with test buffer . ITC measurements of GLP inhibitors binding to the GLP were recorded at 25\u2009\u00b0C using a MicroCal PEAQ-ITC microcalorimeter in the test buffer. The titrations were performed using 10\u2009\u03bcM GLP solution in the sample cell and 100\u2009\u03bcM inhibitor solution in the syringe. A typical experiment consisted of 19 injections under the following parameters: one injection of 0.4\u2009\u03bcL followed by 18 injections of 2.0\u2009\u03bcL; 150\u2009s spacing between injections; 750\u2009rpm stirring speed; and reference power set to 5\u2009\u03bccal s2SO4 (1N). The absorbance at 450\u2009nm of the wells, to which detection reagents were added, was measured in a chemiluminescence reader (ARVO X3 Multilabel Plate Reader), and the values of % inhibition were calculated from the absorbance readings of inhibited wells relative to those of control wells.The tie-2 activity assay was carried out using human recombinant active enzyme and Universal Tyrosine Kinase Assay Kit . The inhibitors were incubated in the presence of 40\u2009mM ATP and 20\u2009ng of tie-2 in the supplied buffer including 5\u2009mM DTT on the microwells of the supplier . After the enzymatic reaction, the mixtures were removed from the wells and all wells were washed four times with phosphate buffered saline with Tween 20 . Then, the wells were blocked (30\u2009min) with blocking buffer, before 50\u2009\u03bcL of anti-phosphated Tyrosine-POD were added to the wells, and incubation was continued . The wells were probed, washed four times with PBS-T (100\u2009\u03bcL), and subsequently incubated with HRP substrate solution . Finally, the reaction was stopped by addition of aqueous HSupplementary Information"} +{"text": "Correction to: Biol Direct (2019) 14:14https://doi.org/10.1186/s13062-019-0247-8After publication of this article , the aut0 \u2013 f \u03c4R0 - f\u03b4\u03c42 ] + f \u03c4 [S(t) + \u03c4\u03b4] (t < 80 years), rather than C = exp (\u03c4 \u2212t/ [C0 \u2013 f \u03c4R0 -f\u03b4\u03c42] + f \u03c4 [S(t) + \u03c4\u03b4] (t < 80 years) in the previous publication.The first error is in the equation of the footnote 1. The correct equation should be: C = exp(-t/ \u03c4) [CThe second error is in the beginning of the third line from the bottom of the footnote currently reads dC/dt = S(t) - C/ \u03c4 whereas it should read dC/dt = f S(t) - C/ \u03c4.The last error is the year in the last sentence of the response to\u00a0Reviewer\u2019s report 2:\u00a0it should be 2018 rather than 1918."} +{"text": "NO2\u22ef\u03c0(N)NO2 inter\u00adaction.The title tetra\u00adnuclear cluster contains a tetra\u00adhedral arrangement of copper(II) ions bonded to a central oxygen atom. The extended structure shows short O\u22efN inter\u00adactions between the nitro groups of adjacent clusters, which are oriented perpendicular to each other in a manner that has previously been described as an O 4Cl6O(C6H9N3O3)4] or [Cu4Cl6O\u00ad(MET)4] [MET is 1-(2-hy\u00addroxy\u00adeth\u00adyl)-2-methyl-5-nitro-1H-imidazole or metronidazole], contains a tetra\u00adhedral arrangement of copper(II) ions. Each copper atom is also linked to the other three copper atoms in the tetra\u00adhedron via bridging chloride ions. A fifth coordination position on each metal atom is occupied by a nitro\u00adgen atom of the monodentate MET ligand. The result is a distorted CuCl3NO trigonal\u2013bipyramidal coordination polyhedron with the axial positions occupied by oxygen and nitro\u00adgen atoms. The extended structure displays O\u2014H\u22efO hydrogen bonding, as well as unusual short O\u22efN inter\u00adactions [2.775\u2005(4)\u2005\u00c5] between the nitro groups of adjacent clusters that are oriented perpendicular to each other. The scattering contribution of disordered water and methanol solvent mol\u00adecules was removed using the SQUEEZE procedure in PLATON .The title tetra\u00adnuclear copper complex, [CuSpek 2015. Acta CrSpek 2009. Acta Cr In the title complex, \u03b1 = 110.80\u2005(9)\u00b0 and \u03b2 = 109.55\u2005(9)\u00b0, such that \u03c44 is 0.990, which indicates negligible deviation from a tetra\u00adhedral geometry for oxygen to 2.4435\u2005(9)\u2005\u00c5 . Each copper atom is also bound to a nitro\u00adgen atom of a MET ligand. The Cu\u2014N lengths range from 1.949\u2005(2) to 1.972\u2005(3)\u2005\u00c5 . Thus, each copper atom sits within a trigonal\u2013bipyramidal arrangement, with the oxygen and nitro\u00adgen atoms forming the axial coordination points, and the bridging chloride ligands occupying the equatorial plane. The trigonal\u2013bipyramidal structure is somewhat distorted, as indicated by the fact that the O\u2014Cu\u2014N angles are less than 180\u00b0, ranging from 173.12\u2005(10) to 176.91\u2005(10)\u00b0 , and the Cl\u2014Cu\u2014Cl angles differ significantly from 120\u00b0, ranging from 109.97\u2005(3) to 134.02\u2005(3)\u00b0 . Furthermore, the O\u2014Cu\u2014Cl angles are all less than 90\u00b0, ranging from 83.33\u2005(6) to 86.13\u2005(6)\u00b0 , indicating that the equatorial chloride ligands are displaced slightly more towards the axial oxygen atom in the center of the mol\u00adecule, than towards the nitro\u00adgen-containing ligand in the opposite axial position.Each of the four copper atoms is linked to the other three copper atoms 5 geometry index is a general descriptor of five-coordinate mol\u00adecules and provides a way to determine the extent of distortion of a mol\u00adecule from trigonal bipyramidal to square pyramidal /60, where \u03b2\u00a0\u2212\u00a0\u03b1 is the difference between the two largest angles , 0.74 (Cu2), 0.84 (Cu3) and 0.73 (Cu4) for the five-coordinate copper centers, giving an average \u03c45 value of 0.74. The \u03c45 values obtained indicate that the copper-centered structures are closer to an idealized trigonal\u2013bipyramidal (1.00) than a square-pyramidal geometry (0.00).The \u03c4A and O21\u2014H21A probably form links to the disordered solvent mol\u00adecules removed with SQUEEZE . The most inter\u00adesting observation is the existence of short O\u22efN inter\u00adactions between the N13/O12/O13 and N33/O32/O33 nitro groups of adjacent clusters that are oriented perpendicular to each other, as illustrated in Fig.\u00a03NO2\u22ef\u03c0(N)NO2 inter\u00adaction 4] contains Cu\u2014X distances that are similar to those in [Cu4Cl6O(imidazole)4] 4] are 1.8960\u2005(18)\u20131.913\u2005(2)\u2005\u00c5, compared to 1.903\u2005(4)\u20131.924\u2005(4)\u2005\u00c5 for [Cu4Cl6O(imidazole)4]. Likewise, the Cu\u2014Cl distances in [Cu4Cl6O(MET)4] are 2.3579\u2005(10)\u20132.4435\u2005(9)\u2005\u00c5, compared to 2.374\u2005(2)\u20132.564\u2005(2)\u2005\u00c5 for [Cu4Cl6O(imidazole)4]. Moreover, the Cu\u2014N distances in [Cu4Cl6O(MET)4] are 1.949\u2005(2)\u20131.972\u2005(3)\u2005\u00c5, compared to 1.934\u2005(6)\u20131.961\u2005(6)\u2005\u00c5.The title compound [CuAnhydrous copper(I) chloride was mixed with MET in methanol (2\u2005ml) in a glass vial, forming a dark olive-colored solution. After allowing the solution to evaporate for eight days, gold-colored plates, suitable for X-ray diffraction, were obtained.Uiso(H) = 1.2Ueq(Csp2) or 1.5Ueq(Csp3). Atoms C34, C35 and O31 and their attached H atoms were modeled as disordered over two sets of sites in a 0.515\u2005(19):0.485\u2005(19) ratio. The structure contains two methanol mol\u00adecules and one water mol\u00adecule, but they are disordered and were removed by the SQUEEZE procedure in PLATON only refer to the main mol\u00adecule.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019008570/hb7801sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019008570/hb7801Isup2.hklStructure factors: contains datablock(s) I. DOI: 1923275CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, 1,3-dioxolane and 1,3-dioxole rings adopt envelope forms, while amide moiety and benzene ring are essentially planar. An intra\u00admolecular O\u2014H\u22efO hydrogen bond supports the mol\u00adecular conformation, and inter\u00admolecular weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions connect the mol\u00adecules into sheet structure. 20H27NO6, the amide moiety is essentially planar, with a maximum deviation of 0.073\u2005(3)\u2005\u00c5, and one of the N-methyl groups shows rotational disorder. The five-membered 1,3-dioxolane ring adopts an envelope form, with the C atom bonded to the olefin side chain as the flap, which deviates from the mean plane through the other four atoms by 0.564\u2005(7)\u2005\u00c5. The 1,3-dioxole ring fused to the benzene ring adopts a flattened envelope form, with the C atom between the two O atoms as the flap, which deviates from the mean plane through the other four atoms by 0.215\u2005(7)\u2005\u00c5. The C\u2014C=C\u2014C olefin moiety is essentially planar and makes a dihedral angle of 87.1\u2005(3)\u00b0 with the benzene ring. An intra\u00admolecular O\u2014H\u22efO hydrogen bond supports the mol\u00adecular conformation, enclosing an S(11) graph-set motif. In the crystal, inter\u00admolecular C\u2014H\u22efO hydrogen bonding links the mol\u00adecules into a tape running along the b axis. Furthermore, other weak C\u2014H\u22efO hydrogen bonds and a C\u2014H\u22ef\u03c0 inter\u00adaction connect the tapes into a sheet structure parallel to (100).In the title compound, C These conversions are often carried out with protection of the contiguous diol shows rotational disorder over two orientations, with refined occupancies of 0.54\u2005(8) and 0.46\u2005(8). The 1,3-dioxolane ring (C10/O11/C12/O13/C14) adopts an envelope form, with puckering parameters of Q(2) = 0.362\u2005(5)\u2005\u00c5 and \u03c6(2) = 40.6\u2005(7)\u00b0. The flap atom C10 deviates from the mean plane through the other four atoms by 0.564\u2005(7)\u2005\u00c5. The 1,3-dioxole ring (C23/C24/O25/C26/O27) in benzodioxole adopts a flattened envelope form, with puckering parameter of Q(2) = 0.135\u2005(5)\u2005\u00c5 and \u03c6(2) = 326\u2005(2)\u00b0. The flap atom C26 deviates from the mean plane through the other four atoms by 0.215\u2005(7)\u2005\u00c5. The olefin moiety (C7\u2014C8=C9\u2014C10) is essentially planar and makes a dihedral angle of 87.1\u2005(3)\u00b0 with the benzene ring (C19\u2013C24). An intra\u00admolecular O\u2014H\u22efO hydrogen bond graph-set motif.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01i; symmetry code as in Table\u00a01b axis, with a C(3) graph-set motif. Furthermore, other weak C\u2014H\u22efO hydrogen bonds and a C\u2014H\u22ef\u03c0 inter\u00adaction , similar to the title compound , gives two entries with refcodes YERGUX , has yet been reported.On the other hand, searching the CSD for a structure with a combination of benzodioxole and oxymethyl\u00addioxolane components, . HRMS (ESI) m/z calculated for C20H27NO6Na+ [M + Na]+: 400.1736; found: 400.1731.The title compound was synthesized in two steps from 3,4-d-arabinose. The H atoms on one of the N-methyl groups (C4) are disordered; they were split into two sets of positions H4A\u2013C and H4D\u2013F, the refined occupancies being 0.54\u2005(8) and 0.46\u2005(8), respectively. C-bound H atoms were positioned geometrically, with C\u2014H = 0.95\u20131.00\u2005\u00c5, and constrained to ride on their parent atoms, with Uiso(H) = 1.5Ueq(C) for methyl groups or 1.2Ueq(C) otherwise. The hy\u00addroxy H atom was placed in a difference map and treated as riding, with O\u2014H = 0.84\u2005\u00c5 and Uiso(H) = 1.5Ueq(O). One problematic reflection global, I. DOI: 10.1107/S2056989018007132/is5496Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018007132/is5496Isup3.cmlSupporting information file. DOI: 1842600CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "These are packed via C\u2014H \u22efN and C\u2014H\u22ef\u03c0 inter\u00adactions, resulting in a three-dimensional architecture with a tilted herringbone packing mode.The two independent mol\u00adecules in the asymmetric unit of the title compound are connected 14H12N4S, was synthesized by the condensation reaction of hydralazine and 2-acetyl\u00adthio\u00adphene and during the reaction, a proton transfer from the imino nitro\u00adgen atom to one of the endocylic nitro\u00adgen atoms occurred. The compound crystallizes in the monoclinic crystal system with two independent mol\u00adecules (mol\u00adecules 1 and 2) in the asymmetric unit. In each mol\u00adecule, there is a slight difference in the orientation of the thio\u00adphene ring with respect to phthalazine ring system, mol\u00adecule 1 showing a dihedral angle of 42.51\u2005(1)\u00b0 compared to 8.48\u2005(1)\u00b0 in mol\u00adecule 2. This implies an r.m.s deviation of 0.428\u2005(1)\u2005\u00c5 between the two mol\u00adecules for the 19 non-H atoms. The two independent mol\u00adecules are connected via two N\u2014H\u22efN hydrogen bonds, forming dimers which inter\u00adact by two bifurcated \u03c0\u2013\u03c0 stacking inter\u00adactions to build tetra\u00admeric motifs. The latter are packed in the ac plane via weak C\u2014H\u22ef\u03c0 inter\u00adactions and along the b axis via C\u2014H \u22efN and C\u2014H\u22ef\u03c0 inter\u00adactions. This results a three-dimensional architecture with a tilted herringbone packing mode.The title compound, C The lengths of the N4\u2014C9 and N4B\u2014C9B bonds of 1.294\u2005(2) and 1.296\u2005(2)\u2005\u00c5, respectively, are in agreement with that of an N=Csp2 bond (1.282 \u00b10.060)\u2005\u00c5 found in the CSD \u2005\u00c5] and N3B\u2014C8B [1.309\u2005(2)\u2005\u00c5] and the single-bond character of N3\u2014N4 [1.398\u2005(2)\u2005\u00c5] and N3B\u2014N4B [1.400\u2005(2)\u2005\u00c5]. Indeed, these values are in agreement with the bond lengths for C=N and N-N bonds in the C=N\u2014N fragment with a cyclic carbon atom and acyclic nitro\u00adgen atoms for organic compounds in the CSD. Such a proton transfer has been reported in other hydrazinophthal\u00adazine derivatives are in a planar conformation with a maximum deviation of 0.006\u2005(1)\u2005\u00c5 for S1 in mol\u00adecule 1 and of 0.003\u2005(1)\u2005\u00c5 for S1via two N\u2014H\u22efN hydrogen bonds .Mol\u00adecules 1 and 2 are linked on Fig.\u00a04 and in tns Fig.\u00a05. The reset al., 2016et al., 2007et al., 2014aA search of the Cambridge Structural Database and hydralazine hydro\u00adchloride in 20\u2005ml of methanol and 10\u2005ml of aqueous solution of sodium acetate as buffering agent. The mixture was refluxed at 338\u2005K under stirring for four\u2005h. The product was left overnight to cool. The yellow precipitate was filtered off and washed several times with water and methanol, and finally crystallized from a mixture of DMF/methanol (2:1) as yellow crystals (in a yield of around 80%) suitable for single-crystal X-ray diffraction studies.Uiso(H)= 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019000732/lh5889sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019000732/lh5889Isup2.cmlSupporting information file. DOI: 1891271CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "R2\u2217 values for 1.5\u2009T and 3\u2009T are reversed. The corrected equation is as follows:In the article titled \u201cHepatic Iron Quantification on 3 Tesla (3\u2009T) Magnetic Resonance (MR): Technical Challenges and Solutions\u201d , there w"} +{"text": "II complex with a Schiff base ligand formed in situ from cyste\u00adamine (2-amino\u00adethane\u00adthiol) and o-vanillin is reported as well results of its anti\u00adbacterial activity screening.The crystal structure of a trinuclear Ni 3(C10H11NO2S)3]\u00b7C3H7NO, with a Schiff base ligand formed in situ from 2-amino\u00adethane\u00adthiol and o-vanillin crystallizes in the ortho\u00adrhom\u00adbic space group Pbca. Its asymmetric unit consists of one neutral Ni3L3 mol\u00adecule and one DMF solvent mol\u00adecule. The solid-state organization of the complex can be described as an insertion of the solvent mol\u00adecules within the crystallographically independent trinuclear NiII species. Several C\u2014H\u22ef\u03c0 edge-to-face inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions link the components in the crystal. A first example of a short inter\u00admolecular C\u2014H\u22efNi contact is found. Anti\u00adbacterial in vitro screening revealed that the title compound has anti\u00adbacterial activity, the best effect being against Acinetobacter baumannii.The title trinuclear nickel(II) complex, [Ni The deviation of the NiII atom from the NOS2 mean plane is 0.0927\u2005(14)\u2005\u00c5. Thus, the mol\u00adecule has a \u2018crown\u2019 or bowl shape with the Ni3S3 unit as its base in a distorted chair conformation. The torsion angles [between 78.49\u2005(5) and 84.79\u2005(5)\u00b0] deviate significantly from the ideal chair conformation for c-hexane which has torsion angles of 60\u00b0. For the Ni2 atom in this core, one additional short contact should be noted, C27\u2014H27C\u22efNi with an H\u22efNi distance of 2.58\u2005\u00c5. Thus, with this additional contact, the coordination geometry of the Ni2 atom is square pyramidal with heteroatom\u2014Ni2\u2014H27C bond angles in the range 78.0-95.1\u00b0.The coordination geometry around each Niet al., 2001et al., 1988Unlike in closely related compounds, the solvent mol\u00adecule is not encapsulated. The distances observed between the Ni atoms are within Ni1\u22efNi2 3.5706\u2005(4) and 3.6656\u2005(5)\u2005\u00c5. The intra\u00admolecular Ni\u2014S distances with the dianionic Schiff base ligand are in the range 2.1888\u2005(12)\u20132.2036\u2005(13)\u2005\u00c5. They are slightly shorter than analogous ones with the bridging sulfur atom of the neighboring ligand [2.2171\u2005(12)\u20132.2262\u2005(13)\u2005\u00c5]. These data are comparable with those previously reported for related structures of 3.722\u2005(6)\u2005\u00c5 for connect the neighboring units . Several C\u2014H\u22efO and C\u2014H\u22ef\u03c0 edge-to-face inter\u00adactions imino\u00admeth\u00adyl]phenolato}trinickel and tris\u00ad(\u03bc2-2-(2-naph\u00adth\u00adyl\u00admeth\u00adoxy)-6-[{(2-sulfido\u00adeth\u00adyl)imino]\u00admeth\u00adyl}phenolato)trinickel(II) di\u00adchloro\u00admethane solvate, tris\u00ad(\u03bc2-2-(benz\u00adyloxy)-6-{[(2-sulfido\u00adeth\u00adyl)imino]\u00admeth\u00adyl}phenolato)tri\u00adnickel(II) di\u00adchloro\u00admethane solvate, tris\u00ad(\u03bc2-2-eth\u00adoxy-6-[{(2-sulfido\u00adeth\u00adyl)imino]\u00admeth\u00adyl}phenolato)trinickel(II) C60-fullerene dichloro\u00admethane solvate, tris\u00ad(\u03bc2-2-eth\u00adoxy-6-{[(2-sulfido\u00adeth\u00adyl)imino]\u00admeth\u00adyl}phenolato)trinickel(II) di\u00adchloro\u00admethane solv\u00adate in ethanol (5\u2005ml) was added to the filtrate and stirred on air magnetically for 2\u2005h. Nickel acetate tetra\u00adhydrate in ethanol (6\u2005ml) was added to the yellowish solution of the Schiff base formed in situ, and the resulting deep-brown solution was stirred magnetically and heated at 340\u2013347\u2005K for 1.5\u2005h resulting in a dark-colored precipitate. The product was isolated by filtration, washed with dry ii=PrOH and finally dried in vacuo. Crystals suitable for crystallographic study were grown from a saturated solution in DMF (deep-brown solution). The crystals were filtered off, washed with dry i-PrOH and finally dried at room temperature (yield: 47%).A solution of KOH in a minimum amount of methanol (2\u20133\u2005ml) was added to a solution of 2-amino\u00adethane\u00adthiol hydro\u00adchloride in methanol (5\u2005ml) and stirred in an ice bath for 10\u2005min. The white precipitate of solid KCl was removed by filtration and \u22121 indicating the formation of a Schiff base (\u2013H\u2014C=N\u2013) (as well in CH3 groups of the solvent mol\u00adecule) and aromatic \u2013C=C\u2013 stretching vibrations. Other strong bands at 1228 and 1244\u2005cm\u22121 are due to the phenolic CO stretching proton signal. The O\u2014CH3 protons peaks only appear at 3.92 ppm. The multiplets of the aromatic protons appear in the range 6.39\u20136.79 ppm with different multiplicity and coupling constants. The strong singlet at 3.39 ppm could be assigned to the aliphatic \u2013CH2\u2013CH2\u2013 protons according to its integral intensity. Signals from the DMF methyl protons appear at 2.94 and 2.78 ppm. Analysis calculated for for C33H40N4Ni3O7S3 (877.00): C, 45.20; H, 4.60; N, 6.39; found: C, 45.5; H, 4.77; N, 6.25.The IR spectrum of the title compound (as KBr pellets) is consistent with the above structural data. It displays the characteristic peak at 1610\u2005cmUiso(H) = 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a02in vitro screening of all test compounds was carried out against reference strains of bacteria and clinical strains . The broth microdilution method was used according to the European Committee on Anti\u00admicrobial Susceptibility Testing (EUCAST). The results obtained indicate that the synthesized compound possesses a broad spectrum of activity against the tested microorganisms and shows relatively better activity against Gram-negative than Gram-positive bacteria. The title complex showed activity with lowest minimum inhibitory concentrations (MIC) values 312.5\u2005\u00b5g ml\u22121 against Gram-negative bacteria E. coli, K. pneumoniae and P. aeruginosa. The highest activity was against clinical strain A. baumannii - MIC = 156.2\u2005\u00b5g ml\u22121. The poorest activity of the complex was against clinical strain Staphylococcus aureus (MRSA). It is well known that A. baumannii is one of the most important nosocomial pathogens because of its longevity in the hospital environment and ability to resist various anti\u00admicrobial agents, such as resistance to broad-spectrum \u03b2-lactam anti\u00adbiotics by \u03b2-lactamases production I. DOI: 10.1107/S2056989019004730/ex2019Isup2.hklStructure factors: contains datablock(s) I. DOI: 1865532CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title di\u00adepoxy\u00adphenalene derivative, two di\u00adhydro\u00adfuran and two tetra\u00adhydro\u00adfuran rings, as well as one cyclo\u00adhexane ring, are fused together with two methyl carboxyl\u00adate groups in positions 2- and 3-. In the crystal, two pairs of C\u2014H\u22efO hydrogen bonds link the mol\u00adecules to form inversion dimers, enclosing two 17H18O6, comprises a fused cyclic system containing four five-membered rings (two di\u00adhydro\u00adfuran and two tetra\u00adhydro\u00adfuran) and one six-membered ring (cyclo\u00adhexa\u00adne). The five-membered di\u00adhydro\u00adfuran and tetra\u00adhydro\u00adfuran rings adopt envelope conformations, and the six-membered cyclo\u00adhexane ring adopts a distorted chair conformation. Two methyl carboxyl\u00adate groups occupy adjacent positions (2- and 3-) on a tetra\u00adhydro\u00adfuran ring. In the crystal, two pairs of C\u2014H\u22efO hydrogen bonds link the mol\u00adecules to form inversion dimers, enclosing two R22(6) ring motifs, that stack along the a-axis direction and are arranged in layers parallel to the bc plane.The title di\u00adepoxy\u00adphenalene derivative, C Reactions totally depending on thermodynamic and kinetic control are infrequently found in the field of organic synthesis, at the same time such transformations are very perspective and attractive from a practical point of view since they allow the direction of the reaction to be changed radically by varying only one of the reaction parameters .et al., 2010et al., 2000et al., 1998et al., 1996et al., 2014et al., 1996et al., 2010et al., 2010et al., 1978The first example of kinetic/thermodynamic control in the course of the Diels\u2013Alder reaction was reported in 1948 into the \u2018domino-adduct\u2019 (2) cyclo\u00adaddition in an inter\u00admediate, leading to the formation of the thermodynamically controlled \u2019domino-adduct\u2019 (2) in an almost qu\u00adanti\u00adtative yield.The present paper describes the uncommon thermal rearrangement of the \u2018 al. 2018 and Bori al. 2018; for ref al. 2018, Lautens al. 2018 and Domi al. 2000]. The tr2) is illustrated in Fig.\u00a01Q(2) = 0.5230\u2005(18)\u2005\u00c5 and \u03c6(2) = 178.1\u2005(2)\u00b0 for ring A, Q(2) = 0.5492\u2005(17)\u2005\u00c5 and \u03c6(2) = 182.3\u2005(2)\u00b0 for B, Q(2) = 0.5230\u2005(18)\u2005\u00c5 and \u03c6(2) = 1.0\u2005(2)\u00b0 for C, and Q(2) = 0.5303\u2005(17)\u2005\u00c5 and \u03c6(2) = 358.9\u2005(2)\u00b0 for D. The puckering parameters of the six-membered cyclo\u00adhexane ring (C1/C2/C10\u2013C13) are QT = 0.518\u2005(2)\u2005\u00c5, \u03b8 = 6.9\u2005(2)\u00b0 and \u03c6 = 178.2\u2005(18)\u00b0. In positions 2- and 3-, i.e. on atoms C8 and C9 . The contribution from the O\u22efH/H\u22efO contacts, corresponding to C\u2014H\u22efO inter\u00adactions, is represented by a pair of sharp spikes characteristic of a strong hydrogen-bonding inter\u00adaction and O\u22efO . The large number of H\u22efH and O\u22efH/H\u22efO inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing and (2), viz. 2-benzyl-6a,9b-bis\u00ad(tri\u00adfluoro\u00admeth\u00adyl)-2,3,6a,9b-tetra\u00adhydro-1H,6H,7H-3a,6:7,9a-di\u00adepoxy\u00adbenzo[de]iso\u00adquinoline -2--2,3-di\u00adhydro-1H,6H,7H-3a,6:7,9a-di\u00adepoxy\u00adbenzo[de]iso\u00adquinoline-3a1,6a-di\u00adcarboxyl\u00adate . C\u2014H\u22ef\u03c0 inter\u00adactions are also observed, together with intra\u00admolecular F\u22efF contacts. The asymmetric unit of HENLEU contains two independent mol\u00adecules. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds, forming columns along [010]. Likewise, C\u2014H\u22ef\u03c0 inter\u00adactions and F\u22efF intra\u00admolecular contacts are also present. In the crystal structure of LIRKAB, inter\u00admolecular C\u2014H\u22efO inter\u00adactions involving the O atoms of the carbonyl groups, the oxygen bridgehead atoms and the meth\u00adoxy O atoms, as well as C\u2014H\u22efF hydrogen bonds, define the crystal packing. These packing features lead to the formation of a supra\u00admolecular three-dimensional structure. C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions are not observed, but H\u22efH inter\u00adactions dominate in the packing. This situation is similar to that in the crystal of the title compound.2) is illustrated in the Scheme. Compound (1) was dissolved in dry o-Me2C6H4 (15\u2005ml) and then heated under reflux for 4\u2005h at \u223c413\u2005K (thin-layer chromatography monitoring). The reaction mixture was cooled and the solvent removed under reduced pressure. The residue was purified by recrystallization from an EtOAc/hexane mixture (1:1 v/v) to give compound (2) as large colourless prismatic crystals . 1H NMR : \u03b4 6.43 , 6.27 , 5.09 , 4.88 , 3.78 , 3.73 , 2.23\u20132.17 , 2.00\u20131.88 1.71\u20131.68 . 13C NMR : \u03b4 164.7 (CO2Me), 162.6 (CO2Me), 150.6 (C-3), 143.8 (C-2), 140.8 (C-7), 138.5 (C-8), 89.3 (C-3a), 85.8 (C-6a), 81.3 (C-1), 80.5 (C-9), 52.2 (C-9a), 52.0 (2 \u00d7 CO2Me), 49.8 (C-9b), 26.7 (C-9), 25.0 (C-6), 17.2 (C-5). IR \u03bdmax/cm\u22121 (KBr): 1709, 1628, 1284, 1261. HRMS (ESI\u2013TOF): calculated for C17H18O6 [M + H]+ 318.1103; found 318.1125.The synthesis of the title compound (Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989019003499/rz5253sup1.cifCrystal structure: contains datablock(s) 2, Global. DOI: 1902671CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, one mol\u00adecule inter\u00adacts with six neighbouring mol\u00adecules 17H14O, which is used as a sensitiser for thermal paper, has a twisted conformation with a dihedral angle of 48.71\u2005(12)\u00b0 between the phenyl ring and the naphthyl ring system. In the crystal, one mol\u00adecule inter\u00adacts with six neighbouring mol\u00adecules via inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions to form a herringbone mol\u00adecular arrangement.The title compound [systematic name: 2-(benz\u00adyloxy)naphthalene], C Herein, we report the crystal structure of 1 as fundamental data for the investigation of its influence on the solid-state physicochemical properties of the thermosensitive layer of the thermal paper.Thermal printing is a rapid and inexpensive printing technology widely used in commercial applications such as receipts, faxes and tickets and the phenyl ring (C12\u2013C17) is 48.71\u2005(12)\u00b0. The related torsion angles for this dihedral angle are \u221244.9\u2005(3)\u00b0 (O1\u2014C11\u2014C12\u2014C17), 178.7\u2005(2)\u00b0 (C1\u2014O1\u2014C11\u2014C12) and \u22125.6\u2005(3)\u00b0 (C6\u2014C1\u2014O1\u2014C11).The title compound Fig.\u00a01 is a simvia inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions = 1.2Ueq(C).The crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019000690/is5508sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019000690/is5508Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019000690/is5508Isup3.cmlSupporting information file. DOI: 1890872CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The two benzene rings are twisted by angles of 79.14\u2005(7) and 19.02\u2005(14)\u00b0 in the two independent mol\u00adecules. In the crystal, amide\u2013amide inter\u00adactions link the mol\u00adecules into chains running along 17H16N4O6, contains two independent mol\u00adecules (A and B). The two benzene rings are twisted by an angle of 79.14\u2005(7)\u00b0 in mol\u00adecule A, whereas, in mol\u00adecule B, they are inclined by 19.02\u2005(14)\u00b0. The conformations of the mol\u00adecules are stabilized by intra\u00admolecular N\u2014H\u22efO hydrogen bonds between the amide nitro\u00adgen atom and the O atom of the ortho-nitro substituent on the phenyl ring, enclosing an S(6) ring motif. In the amide and aliphatic segments, all the N\u2014H, C=O and C\u2014H bonds are anti to each other. In the crystal, the A and B mol\u00adecules are linked by inter\u00admolecular amide-to-amide N\u2014H\u22efO hydrogen bonds, resulting in chains running along the b-axis direction. The inter\u00admolecular inter\u00adactions were analysed using Hirshfeld surface analysis. The two-dimensional fingerprint plots of the inter\u00admolecular contacts indicate that the major contributions are from H\u22efH and O\u22efH inter\u00adactions.The asymmetric unit of the title compound, C There has been a study on the influence of the length of the connecting chain on the anti\u00admalarial activity of bis\u00adquinolines and \u2212161.9\u2005(3)\u00b0 for C2\u2014C1\u2014N1\u2014C7 and C19\u2014C18\u2014N5\u2014C24, respectively. In the other half, they are syn to the ortho-substituent as shown by the torsion angles of 48.6\u2005(4) and \u221250.6\u2005(4)\u00b0 for C13\u2014C12\u2014N2\u2014C11 and C30\u2014C29\u2014N6\u2014C28, respectively. The O1\u2014C7, O2\u2014C11, O7\u2014C24 and O8\u2014C28 bond lengths are 1.213\u2005(3), 1.224\u2005(3), 1.218\u2005(3) and 1.218\u2005(3)\u2005\u00c5, respectively, which indicate that the mol\u00adecules exist in their keto forms in the solid state. In mol\u00adecule A, the bis-amide group forms dihedral angles of 24.79\u2005(12) and 55.04\u2005(7)\u00b0 with the phenyl rings C1\u2013C6 and C12\u2013C17, respectively. In mol\u00adecule B, the plane of the amide group forms dihedral angles of 34.24\u2005(13) and 24.27\u2005(12)\u00b0 with the C18\u2013C23 and C29\u2013C34 phenyl rings, respectively, while the two benzene rings form a dihedral angle of 79.14\u2005(7) and 19.02\u2005(14)\u00b0 in mol\u00adecules A and B, respectively. The planes of mol\u00adecules A and B are almost coplanar with each other, as is evident from the dihedral angle of only 3.15\u2005(17)\u00b0 between phenyl rings C1\u2013C6 and C18\u2013C23.The asymmetric unit of the title compound (I)ortho-substituted nitro groups attached to the C1/C6 and C18/C23 phenyl rings form short intra\u00admolecular contacts, each of 2.01\u2005(3)\u2005\u00c5, with the nearest amide N atom, forming an N\u2014H\u22efO contact resulting in an S(6) hydrogen bonding motif.The O atoms of the b-axis direction. The oxygen atom of the amide C=O group in mol\u00adecule B forms a bifurcated hydrogen bond with the N\u2014H group of the amide unit and the C\u2014H group of the aliphatic chain of an adjacent mol\u00adecule. The C3\u2014H3 unit of the C1\u2013C6 ring of mol\u00adecule A forms a short inter\u00admolecular contact with the oxygen atom O5 belonging to the nitro group of the C12\u2013C17 phenyl ring of another A mol\u00adecule at position \u2212x, 1\u00a0\u2212\u00a0y, \u2212z. C\u2014H groups of the C12\u2013C17 and C29\u2013C34 phenyl rings form hydrogen bonds with the O atoms of the nitro groups of the C12/C17 and C29/C34 phenyl rings at \u2212x, \u2212y\u00a0+\u00a02, \u2212z\u00a0+\u00a01 and \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01, respectively. A packing diagram of the title compound is shown in Fig.\u00a03In the crystal, the mol\u00adecules are linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds Table\u00a01. An inteCrystalExplorer , with de + di \u223c 2.4\u2005\u00c5 . H\u22efH contacts represent the next largest contribution to the Hirshfeld surfaces (29.2%) and show a distinct pattern with a minimum value of de = di \u223c 1.2\u2005\u00c5 . O\u22efC/C\u22efO and N\u22efH/H\u22efN inter\u00adactions cover only 5.4% and 3.4% of the surface, respectively. Two triangles featuring the C\u22efC contacts contribute 3.2% to the Hirshfeld surfaces, with a minimum (de + di) distance of 3.5\u2005\u00c5 .In the two-dimensional fingerprint plot Fig.\u00a06, di is \u2005\u00c5 Fig.\u00a06b. H\u22efH c\u2005\u00c5 Fig.\u00a06c. O\u22efC/C4% Fig.\u00a06d and 3.4% Fig.\u00a06e of the\u2005\u00c5 Fig.\u00a06f.et al., 2010bcN,N\u2032-bis\u00ad(phen\u00adyl)suberamide . The purity of the compound was checked by TLC and it was characterized by IR spectroscopy. The characteristic absorptions were observed at 3334.9, 1693.5 and 1330.9\u2005cm\u22121 for N\u2014H, C=O and C\u2014N, respectively. 1H NMR : 1.93 to 2.00 , 2.48 , 7.95 , 7.28\u20137.33 , 7.63\u20137.70 , 7.82 , 10.24 . 13C NMR : 20.40, 35.19, 124.38, 124.65, 124.68, 131.71, 133.75, 141.36 and 170.82. Rod-shaped yellow single crystals of the title compound were obtained by slow evaporation of a DMF solution at room temperature.A mixture of glutaric acid (0.2\u2005mol) and thionyl chloride (1.0\u2005mol) was heated for half an hour at 363\u2005K. Then 2-nitro\u00adaniline (0.4\u2005mol) was added dropwise under stirring. The resultant mixture was stirred for 3\u2005h and left standing for 12\u2005h for the completion of the reaction. The product was added to crushed ice. The white precipitate obtained was washed thoroughly with water and then with saturated sodium bicarbonate solution and again with water. It was washed first with 2 Uiso(H) = 1.2Ueq(C). The H atoms of the NH groups were located in a difference map and later restrained to a distance of N\u2014H = 0.86\u2005(2)\u2005\u00c5. They were refined with Uiso(H) = 1.2 Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018013075/zl2738sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018013075/zl2738Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018013075/zl2738Isup3.cmlSupporting information file. DOI: 1578746CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the title 1:2 co-crystal has a half mol\u00adecule of twofold symmetric di\u00adthiodi\u00adbenzoic acid and a full mol\u00adecule of benzoic acid. These are connected into three-mol\u00adecule aggregates via hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) hydrogen bonds. 14H10O4S2\u00b72C7H6O2, comprises half a mol\u00adecule of di\u00adthiodi\u00adbenzoic acid [systematic name: 2-[(2-carb\u00adoxy\u00adphen\u00adyl)disulfan\u00adyl]benzoic acid, DTBA], as the mol\u00adecule is located about a twofold axis of symmetry, and a mol\u00adecule of benzoic acid (BA). The DTBA mol\u00adecule is twisted about the di\u00adsulfide bond [the C\u2014S\u2014S\u2014C torsion angle is \u221283.19\u2005(8)\u00b0] resulting in a near perpendicular relationship between the benzene rings [dihedral angle = 71.19\u2005(4)\u00b0]. The carb\u00adoxy\u00adlic acid group is almost co-planar with the benzene ring to which it is bonded [dihedral angle = 4.82\u2005(12)\u00b0]. A similar near co-planar relationship pertains for the BA mol\u00adecule [dihedral angle = 3.65\u2005(15)\u00b0]. Three-mol\u00adecule aggregates are formed in the crystal whereby two BA mol\u00adecules are connected to a DTBA mol\u00adecule via hy\u00addroxy-O\u2014H\u22efO(hydroxy) hydrogen bonds and eight-membered {\u22efHOC=O}2 synthons. These are connected into a supra\u00admolecular layer in the ab plane through C\u2014H\u22efO inter\u00adactions. The inter\u00adactions between layers to consolidate the three-dimensional architecture are \u03c0\u2013\u03c0 stacking inter\u00adactions between DTBA and BA rings [inter-centroid separation = 3.8093\u2005(10)\u2005\u00c5] and parallel DTBA-hy\u00addroxy-O\u22ef\u03c0(BA) contacts [O\u22efring centroid separation = 3.9049\u2005(14)\u2005\u00c5]. The importance of the specified inter\u00adactions as well as other weaker contacts, e.g. \u03c0\u2013\u03c0 and C\u2014H\u22efS, are indicated in the analysis of the calculated Hirshfeld surface and inter\u00adaction energies.The asymmetric unit of the title 1:2 co-crystal, C Such oxidation of the original 2-thio\u00adbenzoic acid to DTBA is well known in co-crystallization studies i\u2014C3i torsion angle being \u221283.19\u2005(8)\u00b0; symmetry operation (i): 1\u00a0\u2212\u00a0x, y, z. This almost orthogonal disposition is also seen in the dihedral angle between the benzene rings of 71.19\u2005(4)\u00b0. The presence of a carb\u00adoxy\u00adlic acid group is readily confirmed by the disparity in the C1\u2014O1, O2 bond lengths, i.e. 1.317\u2005(2) and 1.229\u2005(2)\u2005\u00c5, respectively. This group is practically co-planar with the benzene ring to which it is bonded, as seen in the dihedral angle of 4.82\u2005(12)\u00b0. This co-planar arrangement allows for a significant intra\u00admolecular S\u2190O inter\u00adaction, i.e. S1\u22efO2 = 2.6712\u2005(12)\u2005\u00c5, as the carbonyl-O2 atom is orientated towards a di\u00adsulfide-S1 atom and 1.233\u2005(2)\u2005\u00c5, respectively. As for the DTBA mol\u00adecule, the carb\u00adoxy\u00adlic acid group is close to co-planar with the benzene ring to which it is bound, forming a dihedral angle of 3.65\u2005(15)\u00b0.PLATON . The resultant three-mol\u00adecule aggregates are connected through DTBA-C\u2014H\u22efO3(hydroxyl-BA) and BA-C\u2014H\u22efO1(hydroxyl-DTBA) inter\u00adactions, to form non-symmetric, ten-membered {O\u22efHCCC}2 homo-synthons leading to supra\u00admolecular layers in the ab plane, Fig.\u00a02b). Owing to the nearly right-angle relationship between the rings in the DTBA mol\u00adecule, and the co-planarity between the carb\u00adoxy\u00adlic acid groups and the respective rings they are connected to, the layers also have a similar topology. Adjacent layers inter-digitate with other layers, on both sides, i.e. approximately orthogonally, as highlighted in Fig.\u00a02c). As illustrated in Fig.\u00a02d), the connections between layers are of two types and include \u03c0\u2013\u03c0 stacking inter\u00adactions between DTBA and BA rings with the inter-centroid (C2\u2013C7)\u22ef(C9\u2013C14)iv separation being 3.8093\u2005(10)\u2005\u00c5, an angle of inclination of 8.36\u2005(8)\u00b0 and an off-set of 1.40\u2005\u00c5 for symmetry operation (iv): 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z. The second inter\u00adaction is a weak, parallel DTBA-hy\u00addroxy-O1\u22ef\u03c0(C9\u2013C14)ii contact with a O1\u22efring centroid(C9\u2013C14)v separation of 3.9049\u2005(14)\u2005\u00c5 and angle at O1 = 60.96\u2005(9)\u00b0 for symmetry operation (v): x, y, 1\u00a0\u2212\u00a0z.The geometric parameters characterizing the inter\u00adatomic contacts, as identified in dnorm) as well as calculation of the inter\u00adaction energies using CrystalExplorer and external (de) distances to the nearest nucleus dnorm mapping of the three-mol\u00adecule aggregate is shown in Fig.\u00a03\u22121, Table\u00a02\u22121. The short \u03c0\u2013\u03c0 inter\u00adaction involving the DTBA and BA benzene rings, mentioned in Supra\u00admolecular features, has an inter\u00adaction energy of \u221221.7\u2005kJ\u2005mol\u22121, i.e. more stable than the C\u2014H\u22efO inter\u00adactions. The energy calculation reveals that such an inter\u00adaction is mainly dispersive in nature, cf Table\u00a02a) and (b), and the mol\u00adecular dimer sustained by \u03c0\u2013\u03c0 contacts in Fig.\u00a04c).The v separation of 4.4323\u2005(10)\u2005\u00c5, an angle of inclination of 8.36\u2005(8)\u00b0 and an off-set of 2.74\u2005\u00c5 for symmetry operation (v): x, y, 1\u00a0\u2212\u00a0z. In addition, a BA-benzene-C6\u2014H\u22efS contact (2.94\u2005\u00c5) is noted, Table\u00a02Other important but less significant contacts are noted through the Hirshfeld surface analysis such as a longer \u03c0\u2013\u03c0 inter\u00adaction between DTBA and BA rings with an inter-centroid (C2\u2013C7)\u22ef(C9\u2013C14)di and de contact distances at the inter\u00adval of 0.01\u2005\u00c5 , O\u22efH/H\u22efO , S\u22efH/H\u22efS and other contacts (8.0%). Among these contacts, only the O\u22efH/ H\u22efO, C\u22efC and S\u22efH/ H\u22efS contacts are shorter than the respective sums of the van der Waals radii to result in meaningful inter\u00adactions in the crystal, i.e. O\u22efH, C\u22efC and S\u22efH = \u223c1.72, \u223c3.4 and \u223c3.0\u2005\u00c5, respectively.A qu\u00adanti\u00adtative analysis of the Hirshfeld surfaces was performed through the generation of two-dimensional fingerprint plots by combining the di + de distances except for the O\u22efH/H\u22efO inter\u00adactions. Thus, overall (I)i.e. 12.8 versus 12.3%. The apparent disparity arises as a result of the larger surface area to volume ratio for the DTBA mol\u00adecule as compared to the DTBA+BA aggregate when it is considered as a single entity, hence leading to greater exposure of the O\u22efH/H\u22efO contacts in DTBA within its surrounding inter\u00adacting environment. A smaller disparity is evident for the -S\u22efH-/-H\u22efS- contacts, in that the former constitutes about 6.3% in (I)A close inspection on the corresponding decomposed fingerprint plots of the individual DTBA and BA mol\u00adecules reveals similar compositions as well as a)\u2013(c), the crystal is significantly governed by electrostatic force owing to the strong O\u2014H\u22efO inter\u00adactions that result in an alternate V-shape energy topology across the b-axis direction. A relatively less significant, but essential dispersion contribution is also observed and arises from the \u03c0\u2013\u03c0 inter\u00adactions spanning all benzene rings. Overall, it can be concluded that these inter\u00adacting forces directed the assembly of the mol\u00adecules in (I)In order to study the overall topology of the energy distributions in the crystal of (I)Structural commentary, the DTBA mol\u00adecule is twisted about the central di\u00adsulfide bond, having a C\u2014S\u2014S\u2014C torsion angle of \u221283.19\u2005(8)\u00b0. A survey of the literature indicates that this is a common feature of such mol\u00adecules. A search of the Cambridge Structural Database ethene solution of the ground mixture. M.p. 384.2\u2013385.6\u2005K. IR (cm\u22121): 3070 \u03bd(C\u2014H), 1677 \u03bd(C=O), 1584 \u03bd(C=C), 1415 \u03b4(C\u2014H), 706 \u03b4(C=C), 684 \u03b4(OCO).All chemicals were of reagent grade and used as received without purification. 2-Thio\u00adbenzoic acid was mixed with benzoic acid and ground for 15 minutes in the presence of a few drops of methanol. The procedure was repeated three times. Colourless blocks were obtained by carefully layering toluene (1\u2005ml) on an Uiso(H) set to 1.2Ueq(C). The oxygen-bound H atoms were located from difference-Fourier maps and refined without constraint.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989018017097/hb7790sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018017097/hb7790Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018017097/hb7790Isup3.cmlSupporting information file. DOI: 1882556CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Z\u2032 = 1 is reported and compared with first polymorph with Z\u2032 = 2.A second polymorph of 3-meth\u00adoxy\u00adbenzoic acid with 8H8O3, is described in the centrosymmetric monoclinic space group P21/c with Z\u2032 = 1 as compared to the first polymorph, which crystallizes with two conformers (Z\u2032 = 2) in the asymmetric unit in the same space group. In the crystal of the second polymorph, inversion dimers linked by O\u2014H\u22efO hydrogen bonds occur and these are linked into zigzag chains, propagating along the b-axis direction by C\u2014H\u22efO links. The crystal structure also features a weak \u03c0\u2013\u03c0 inter\u00adaction, with a centroid-to-centroid distance of 3.8018\u2005(6)\u2005\u00c5. The second polymorph of the title compound is less stable than the reported first polymorph, as indicated by its smaller calculated lattice energy.A new polymorphic form of the title compound, C Polymorph I\u03b1 crystallizes in the monoclinic space group P21/n with a = 13.8034\u2005(17)\u2005\u00c5, b = 5.0275\u2005(5)\u2005\u00c5, c = 21.446\u2005(3)\u2005\u00c5 and \u03b2 = 99.320\u2005(13)\u00b0 , which are connected into a homodimer through strong O\u2014H\u22efO hydrogen bonds. As described below, these two conformers (A and B) differ in the orientation of the meth\u00adoxy group and its relative position from the \u2014OH group. DFT calculations suggest that the A conformer of I\u03b1 is more energetically stable than the B conformer . The mol\u00adecule is almost planar with a maximum deviation of 0.107\u2005(1)\u2005\u00c5 at atom O1. The mol\u00adecules of I\u03b2 adopt a similar conformation (overlay r.m.s.d. = 0.052\u2005\u00c5) as compared to the conformer A of I\u03b1 of I\u03b2 is close to coplanar with the attached phenyl ring (C1\u2013C6) as indicated by the dihedral angle of 5.6\u2005(7)\u00b0. The C8\u2014O3\u2014C3\u2014C2 torsion angle of I\u03b2 is \u2212176.63\u2005(7)\u00b0 as compared to \u2212176.75\u2005(11) and \u22121.4\u2005(2)\u00b0 for conformers A and B, respectively, of I\u03b1.The asymmetric unit of I\u03b2 Fig.\u00a01 consistsb-axis direction. The [010] chains are stacked along the a axis into corrugated sheets parallel to the ab plane via weak \u03c0\u2013\u03c0 inter\u00adactions with a centroid-to-centroid distance of 3.8018\u2005(6)\u2005\u00c5 and slippage of 1.676\u2005\u00c5.In the crystal of I\u03b2, two inversion-related mol\u00adecules are joined into a homodimer with an ds Fig.\u00a02. The homdnorm and the two-dimensional fingerprint plots for I\u03b2 were generated using CrystalExplorer17.5 . The H\u22efH contact is the most populated contact and contributes 42.3% of the total inter\u00admolecular contacts, followed by H\u22efO/O\u22efH (32.9%), H\u22efC/C\u22efH (11.4%) and C\u22efC (8.1%) contacts .The Hirshfeld surfaces mapped with normalized contact distance rm Fig.\u00a03 correspots Fig.\u00a04. The tipPIXEL software is larger than that of I\u03b2 (98.5\u2005kJ\u2005mol\u22121) and this comparison is in agreement with the report of Pereira Silva et al. , see: Parvez 1987 and Ette al. 1988. For the al. 1988, Colapie al. 1988, Fausto al. 1997 and Hath al. 2011. For the al. 2014 and Pere al. 2015.Single crystals of I\u03b2 were obtained from an unsuccessful attempt of co-crystallization between 3-meth\u00adoxy\u00adbenzoic acid and hexa\u00admethyl\u00adene\u00adtetra\u00admine. Colourless plate-like crystals were obtained from slow evaporation of a methano\u00adlic mixture of 3-meth\u00adoxy\u00adbenzoic acid and hexa\u00admethyl\u00adene\u00adtetra\u00admine in equimolar ratio at room temperature.O2 = 1.01\u2005(2)\u2005\u00c5]. The remaining H atoms were positioned geometrically [C\u2014H = 0.95 and 0.98\u2005\u00c5] and refined using a riding model with Uiso(H) = 1.2 or 1.5Ueq(C). A rotating group model (AFIX 137) was applied to the methyl group.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018016900/hb7789sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018016900/hb7789Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018016900/hb7789sup3.docxHirshfeld shape-index and curvedness Figures. DOI: Click here for additional data file.10.1107/S2056989018016900/hb7789Isup4.cmlSupporting information file. DOI: 1448794CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "C5H7N2+\u00b7C6H7O7\u2212 (I) and 3C5H7N2+\u00b7C6H5O73\u2212 (II). Salt I is formed by the protonation of the pyridine N atom and deprotonation of the central carb\u00adoxy\u00adlic group of the acid, while in II all three carb\u00adoxy\u00adlic groups of the acid are deprotonated and the charges are compensated for by three 2-amino\u00adpyridinium cations.2-Amino\u00adpyridine and citric acid mixed in 1:1 and 3:1 ratios in ethanol yielded crystals of two 2-amino\u00adpyridine citric acid salts, viz. C5H7N2+\u00b7C6H7O7\u2212 (I) (systematic name: 2-amino\u00adpyridin-1-ium 3-carb\u00adoxy-2-carb\u00adoxy\u00admethyl-2-hy\u00addroxy\u00adpropano\u00adate), and 3C5H7N2+\u00b7C6H5O73\u2212 (II) . The supra\u00admolecular synthons present are analysed and their effect upon the crystal packing is presented in the context of crystal engineering. Salt I is formed by the protonation of the pyridine N atom and deprotonation of the central carb\u00adoxy\u00adlic group of citric acid, while in II all three carb\u00adoxy\u00adlic groups of the acid are deprotonated and the charges are compensated for by three 2-amino\u00adpyridinium cations. In both structures, a complex supra\u00admolecular three-dimensional architecture is formed. In I, the supra\u00admolecular aggregation results from Namino\u2014H\u22efOacid, Oacid\u22efH\u2014Oacid, Oalcohol\u2014H\u22efOacid, Namino\u2014H\u22efOalcohol, Npy\u2014H\u22efOalcohol and Car\u2014H\u22efOacid inter\u00adactions. The mol\u00adecular conformation of the citrate ion (CA3\u2212) in II is stabilized by an intra\u00admolecular Oalcohol\u2014H\u22efOacid hydrogen bond that encloses an S(6) ring motif. The complex three-dimensional structure of II features Namino\u2014H\u22efOacid, Npy\u2014H\u22efOacid and several Car\u2014H\u22efOacid hydrogen bonds. In the crystal of I, the common charge-assisted 2-amino\u00adpyridinium\u2013carboxyl\u00adate heterosynthon exhibited in many 2-amino\u00adpyridinium carboxyl\u00adates is not observed, instead chains of N\u2014H\u22efO hydrogen bonds and hetero O\u2014H\u22efO dimers are formed. In the crystal of II, the 2-amino\u00adpyridinium\u2013carboxyl\u00adate heterosynthon is sustained, while hetero O\u2014H\u22efO dimers are not observed. The crystal structures of both salts display a variety of hydrogen bonds as almost all of the hydrogen-bond donors and acceptors present are involved in hydrogen bonding.2-Amino\u00adpyridine and citric acid mixed in 1:1 and 3:1 ratios in ethanol yielded crystals of two 2-amino\u00adpyridinium citrate salts, Organic crystals, especially salts, are now considered as potential materials for optical applications because of their flexibility in mol\u00adecular design , whose structure is illustrated in Fig.\u00a01I have different conformations. In one of them (C5/O4/O5) the O\u2014H and C=O bonds are in a syn conformation while in the other (C3/O2/O3), they have an anti conformation hydrogen bonds [acid(t1) = C3/O2/O3], viz. N2\u2014H2D\u22efO2 , 4.762 and 6.396 . Thus, an equimolar mixing of citric acid and 2-amino\u00adpyridine resulted in the formation of salt on Fig.\u00a01. In the 2 Table\u00a01.II, illustrated in Fig.\u00a023\u2212 [(C5H5O7)3\u2212], and three 2-AMP+ cations , wherein the pyridine N atom of each 2-AMP unit is protonated and all three carb\u00adoxy\u00adlic groups of the acid are deprotonated. This is supported by the observation that the C\u2014O bonds of all the three carb\u00adoxy\u00adlic groups have similar bond lengths, in the range 1.231\u2005(2)\u20131.266\u2005(2)\u2005\u00c5, which is an indication of the partial double-bond character of all of the C\u2014O bonds resulting from deprotonation. The mol\u00adecular conformation of the CA3\u2212 anion is stabilized by an intra\u00admolecular Oalcohol\u2014H\u22efOacid(t1) hydrogen bond, namely O1\u2014H1O\u22efO3, that closes an S(6) ring motif hydrogen bond, namely N1\u2014H1B\u22efO6. The second cation, 2-AMP2, inter\u00adacts with the CA3\u2212 anion via a charge-assisted 2-amino\u00adpyridinium-carboxyl\u00adate amino\u2014H\u22efOacid(t1) (N3\u2014H3A\u22efO3) and Npy\u2014H\u22efOacid(t1) (N4\u2014H4\u22efO2) hydrogen bonds. The third cation, 2-AMP3, inter\u00adacts with the anion via a discrete Namino\u2014H\u22efOacid(c) hydrogen bond, namely N6\u2014H6B\u22efO7.In the asymmetric unit of salt I are given in Table\u00a01I, the cations and anions of adjacent units are inter\u00adconnected by a Car\u2014H\u22efOacid(t1) inter\u00adactions, viz. C9\u2014H9\u22efO3, while adjacent anions related by b-glide symmetry form chains running along the b-axis direction, consisting of an acid(c)\u22efH\u2014Oacid(t1) and Oalcohol\u2014H\u22efOacid(c) hydrogen bonds, namely O3\u2014H3\u22efO7i and O1\u2014H1\u22efO6i; see Fig.\u00a03+ and CA\u2212 ions further aggregate to form sheets parallel to the ac plane \u2014H\u22efOacid(c) hydrogen bonds, namely O4\u2014H4\u22efO6iii, running along the a-axis direction and linking the twofold-symmetry-related CA\u2212 anions (Table\u00a01+ ions via Namino\u2014H\u22efOacid(t1)=C hydrogen bonds, namely N2\u2014H2D\u22efO2, and an amino\u2014H\u22efOalcohol and Npy\u2014H\u22efOalcohol hydrogen bonds, N2\u2014H2C\u22efO1ii and N1\u2014H1A\u22efO1ii, respectively, is formed hydrogen bond. The CA3\u2212 anion and the first 2-AMP+ cation (2-AMP1) form sheets lying parallel to the (101) plane . The sheet consists of alternating CA3\u2212 and 2-AMP+ ions, forming chains via C11\u2014H11\u22efO2iii inter\u00adactions, with adjacent anti-parallel chains linked by C10\u2014H10\u22efO2ii, N1\u2014H1A\u22efO7i, N1\u2014H1B\u22efO6, N2\u2014H2\u22efO7i and N2\u2014H2\u22efO1i hydrogen bonds (Table\u00a02+ ions (2-AMP2) propagate alternately along the a-axis direction to form ribbons consisting of alternating A\u22efO3 and N4\u2014H4\u22efO2 hydrogen bonds (Table\u00a02B\u22efO4 and C13\u2014H13\u22efO6 hydrogen bonds (Table\u00a02+ ions (2-AMP3) are inter\u00adlinked to the adjacent citrate ions, forming ribbons of alternating i and N6\u2014H6A\u22efO5i hydrogen bonds . Adjacent ribbons are further inter\u00adconnected by N6\u2014H6B\u22efO7 hydrogen bonds to form corrugated sheets parallel to the ab plane (Table\u00a02b). Overall a complex supra\u00admolecular three-dimensional structure is formed.In the crystal of s Table\u00a02. Howeverne Fig.\u00a05a and 5bs Table\u00a02. On the ns Fig.\u00a06a consiss Table\u00a02 and (11)s Table\u00a02. Finallys Table\u00a02, and . The most common hydrogen bonds observed in these hydrated salts are Namine\u2014H\u22efOcitric, Namine\u2014H\u22efOwater and Owater\u2014H\u22efOcitric, forming different supra\u00admolecular architectures. In the absence of a water mol\u00adecule, the most common hydrogen bonds are Namine\u2014H\u22efOcitric and Ocitric\u2014H\u22efOcitric. However, the nature of these supra\u00admolecular synthons varies from one structure to another, depending on the nature of the organic cations.A survey of the Cambridge Structural Database . Single crystals of salt II were obtained from a similar procedure; an ethano\u00adlic solution (15\u2005ml) of citric acid was mixed with an ethano\u00adlic solution (15\u2005ml) of 2-amino\u00adpyridine .A solution of citric acid in ethanol (15\u2005ml) was added to an ethano\u00adlic solution (15\u2005ml) of 2-amino\u00adpyridine . The resulting solution was heated and the hot solution was filtered. Slow evaporation of the solution resulted in the formation of colourless prismatic crystals of salt I, the OH H atom (H1) was positioned geometrically and refined as riding: O\u2014H = 0.82\u2005\u00c5 with Uiso(H) = 1.5Ueq(O). In salt II, the OH H atom (H1O) was located in a difference-Fourier map and freely refined. In both salts, the other H atoms were positioned geometrically and refined as riding: N\u2014H = 0.86\u2005\u00c5, C\u2014H = 0.93\u20130.97\u2005\u00c5 with Uiso(H) = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018009787/su5449sup1.cifCrystal structure: contains datablock(s) I, II, Global. DOI: 10.1107/S2056989018009787/su5449Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989018009787/su5449IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989018009787/su5449Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018009787/su5449IIsup5.cmlSupporting information file. DOI: 1854628, 1854627CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Deletional hereditary persistence of fetal hemoglobin (HPFH)/\u03b4\u03b2\u2010thalassemia and \u03b4\u2010thalassemia are rare inherited disorders which may complicate the diagnosis of \u03b2\u2010thalassemia. The aim of this study was to reveal the frequency of these two disorders in Southwestern China.2 levels were confirmed by \u03b4\u2010globin gene sequencing. Furthermore, the pathogenicity and construction of a selected \u03b4\u2010globin mutation were analyzed.A total of 33,596 subjects were enrolled for deletional HPFH/\u03b4\u03b2\u2010thalassemia, and positive individuals with high fetal hemoglobin (Hb F) level were diagnosed by multiplex ligation\u2010dependent probe amplification (MLPA). A total of 17,834 subjects were analyzed for mutations in the \u03b4\u2010globin gene. Positive samples with low Hb A2 \u22642.0%) were characterized by molecular analysis. \u03b4\u2010Globin gene mutation was found at a frequency of 0.49% in Yunnan. The pathogenicity and construction for a selected \u03b4\u2010globin mutation was predicted.A total of 92 suspected cases with Hb F \u22655.0% were further characterized by MLPA. Eight different deletional HPFH/\u03b4\u03b2\u2010thalassemia were observed at a frequency of 0.024%. In addition, 195 cases suspected to have a \u03b4\u2010globin gene mutation (Hb AScreening of these two disorders was analyzed in Southwestern China, which could define the molecular basis of these conditions in this population. Fetal hemoglobin (Hb F) is a minor hemoglobin that is composed of two \u03b1\u2010 and two \u03b3\u2010globin chains (\u03b12\u03b32). Hereditary persistence of fetal hemoglobin is caused by mutations in the promoter of the \u03b3\u2010globin gene . As summarized in Globin Gene Server home page (http://globin.cse.psu.edu/hbvar/menu.html), more than 50 deletional HPFH/\u03b4\u03b2\u2010thalassemias and 130 mutations in the \u03b4\u2010globin gene have been reported to date. However, few reports on deletional HPFH/\u03b4\u03b2\u2010thalassemia and mutations in the \u03b4\u2010globin gene in Chinese population have been investigated at the molecular level. The aim of this study was to determine the frequency of deletional HPFH/\u03b4\u03b2\u2010thalassemia mutations and \u03b4\u2010globin gene mutations in Yunnan population. Furthermore, we characterized a rare \u03b4\u2010globin gene mutation using a comprehensive clinical and structure\u2010function analysis. These findings are important for accurate thalassemia prenatal diagnosis, as well as for providing molecular insights into new mutations in \u03b4\u2010globin gene.Both deletional HPFH/\u03b4\u03b2\u2010thalassemia and \u03b4\u2010thalassemia mutations are related to different ethnic backgrounds and (b) low levels of Hb A2 associated with a visible second Hb A2 fraction . Internal quality control of the hemoglobin analysis was performed using the control materials provided by the manufacturer. A total of 33,596 subjects were screened for deletional HPFH/\u03b4\u03b2\u2010thalassemia using capillary electrophoresis from July 2014 to September 2016. Individuals showing Hb F \u22655.0% were considered to be carriers of deletional HPFH/\u03b4\u03b2\u2010thalassemia 0 thalassemia and Southeast Asia HPFH (SEA\u2010HPFH) deletion were identified by Gap\u2010PCR was performed using multiplex ligation\u2010dependent probe amplification (MLPA) following the manufacturer's instructions . Two commonest deletional HPFH, Chinese 2.43.7 (NC_000016.9:g.223300_227103del), \u2010\u03b14.2 (NC_000016.9:g.219817_(223755_224074)del), \u2010\u2010SEA (NC_000016.9:g.215400_234700del), \u03b1CS\u03b1 , \u03b1WS\u03b1 , and \u03b1QS\u03b1 .Two fragments of the \u03b4\u2010globin gene (NG_000007.3) were amplified using the following primers: \u03b41\u2010F 5\u2032CTGAGTCAAGACACACATGACAG3\u2032, \u03b41\u2010R 5\u2032 TGGTATGCATAATTTGAGTTGTTG3\u2032; \u03b42\u2010F 5\u2032 AATATCCTGTCTTTCTCTCCCAAC3\u2032, \u03b42\u2010R 5\u2032 TAATTTCTGCTCTTTGGAGGTAG3\u2032 and SIFT (http://sift.jcvi.org) were determined for the normal and mutant \u03b4\u2010globin variant (monomer) using the Isoelectric Point Calculator (IPC) /\u03b4\u03b2\u2010thalassemia, 165 positive samples were selected (Hb F \u22655.0%). Eight cases of deletional HPFH/\u03b4\u03b2\u2010thalassemia were found by MLPA Figure , with a 0 deletion). Two unrelated individuals (case 3 and case 4) showed a similar (\u03b4\u03b2)0\u2010deletion. A G\u03b3(A\u03b3\u03b4\u03b2)0\u2010deletion was found in case 5. Two unrelated individuals (cases 6 and case 7) had a G\u03b3(A\u03b3\u03b4\u03b2)0 deletion, and this was confirmed as Chinese G\u03b3(A\u03b3\u03b4\u03b2)0 thalassemia by Gap\u2010PCR. Furthermore, case 7 was a compound heterozygous for Chinese G\u03b3(A\u03b3\u03b4\u03b2)0 and IVS\u2010I\u20101 (G\u00a0>\u00a0T) (HBB:c.92\u00a0+\u00a01G>T). Case 8 showed a \u201cdiscontinuous\u201d (\u03b5\u03b3\u03b4\u03b2)0 deletion, where the size of the deletion ranged from OR51M1\u20101 probe to hemoglobin subunit beta (HBB)\u2010up probe; however, among this deletion region, the hemoglobin subunit gamma (HBG)2\u20133 and HBG1\u2010up probes detected a fragment. Therefore, this deletion was considered to be a \u201cdiscontinuous\u201d (\u03b5\u03b3\u03b4\u03b2)0 deletion, and its sequence was further confirmed by targeted next\u2010generation sequencing or a second Hb A2 fraction, and were selected for further molecular diagnosis. Seven types of \u03b4\u2010globin mutations were found in 87 \u03b4\u2010thalassemia patients mutation had Hb A2 values below 1.6% (Hb A2\u00a0\u2264\u00a01.6%). One sample was a compound heterozygote for \u221277 (T\u00a0>\u00a0C) and \u2010\u03b13.7 with Hb A2 value of 1.4% .The number of subjects for each Hb A3.312/L, Hb 13.8g/dl, MCV 91.0fl, MCH 30.3pg, RDW\u2010CV 13.1%, and MCHC 333g/L. The predicted pIs of the wild type and mutant type were 7.42 and 6.87 respectively mutation was selected for further bioinformatics analysis. The proband was a 31\u2010year\u2010old Chinese woman. Her hematological characteristics were as follows: RBC 4.5\u00d710y Figure a. The Lyy Figure b. Polyphy Figure c. Howevey Figure d.2O atoms and B cell CLL/lymphoma 11A (BCL11A) can modulate fetal\u2010to\u2010adult globin switching. Mutations in KLF1 can result in significantly boosted Hb F levels in normal individuals 0\u2010deletion, one case of G\u03b3(A\u03b3\u03b4\u03b2)0\u2010deletion, two cases of Chinese G\u03b3(A\u03b3\u03b4\u03b2)0 deletion, and one case of (\u03b5\u03b3\u03b4\u03b2)0\u2010deletion were identified. All eight samples had typical \u03b2\u2010thalassemia phenotypes with hypochromic and microcytic erythrocytes due to deletions in the \u03b2\u2010globin gene or promoter. Case 7 was a compound heterozygote for the Chinese G\u03b3(A\u03b3\u03b4\u03b2)0 deletion and IVS\u2010I\u20101 (G\u00a0>\u00a0T). As a result, this patient had typical \u03b2\u2010thalassemia intermedia symptoms with increased Hb F levels should be regarded as the frequency of deletional HPFH/\u03b4\u03b2\u2010thalassemia but not \u03b2\u2010globin cluster deletions. Some \u03b2\u2010globin cluster deletions, such as the 118\u00a0kb Filipino deletion with a normal Hb F 1.7% cannot be screened by capillary electrophoresis . The individuals who have \u03b4\u2010globin gene mutations do not usually have clinical phenotypes because of the physiologically lower expression levels of the \u03b4\u2010globin gene Lys\u00a0>\u00a0Asn, HBB:c.198G\u00a0>\u00a0T]. A previous study also had shown that those heterozygous for Hb J Sicilia had no clinically significant problems (Ricco et al., 2 with a slight subunit modification (Sen et al., 2 structure obviously and p.K65N mutation in \u03b4\u2010globin gene should be benign.The 3D models evaluated by SWISS\u2010MODEL and PyMol showed that the K65 was located in a helix near the periphery of the protein. Based on the results from SWISS\u2010MODEL and PyMol, we observed that K65 was not positioned in a complex or important interaction network. Therefore, the substitution of the Lys residue should not significantly disrupt the structure and function of Hb AIn conclusion, this is the first report on the frequency and spectrum of deletional HPFH/\u03b4\u03b2\u2010thalassemia and mutations in the \u03b4\u2010globin gene in a Southwestern Chinese population. Bioinformatic analysis of a rare mutation characterized the potential changes at the protein\u2010level. Our study will provide a guideline for genetic counseling and prenatal diagnosis.The author reports no conflict of interest in this work.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file."} +{"text": "The di\u00adhydro\u00adbenzo\u00addiazole moiety is not quite planar while the whole mol\u00adecule adopts a U-shaped conformation in which there is a close approach of the two benzyl groups. Chains of alternating mol\u00adecules and lattice water extending along the normal to (301) are formed by O\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds. 24H21N5O\u00b7H2O, the di\u00adhydro\u00adbenzo\u00addiazole moiety is not quite planar, while the whole mol\u00adecule adopts a U-shaped conformation in which there is a close approach of the two benzyl groups. In the crystal, chains of alternating mol\u00adecules and lattice water extending along [201] are formed by O\u2014HUncoordW\u22efODhyr and O\u2014HUncoordW\u22efNTrz hydrogen bonds. The chains are connected into layers parallel to (010) by C\u2014HTrz\u22efOUncoordW hydrogen bonds with the di\u00adhydro\u00adbenzo\u00addiazole units in adjacent layers inter\u00adcalating to form head-to-tail \u03c0-stacking [centroid-to-centroid distance = 3.5694\u2005(11)\u2005\u00c5] inter\u00adactions between them, which generates the overall three-dimensional structure. Hirshfeld surface analysis indicates that the most important contributions for the crystal packing are from H\u22efH (52.1%), H\u22efC/C\u22efH (23.8%) and O\u22efH/H\u22efO (11.2%) inter\u00adactions. Hydrogen-bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Density functional theory (DFT) optimized structures at the B3LYP/ 6\u2013311\u2005G level are compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.In the title mol\u00adecule, C The benzene ring D (C12\u2013C17) is inclined to the triazole ring C by 78.91\u2005(11)\u00b0 while the latter ring is inclined to the B ring by 64.70\u2005(11)\u00b0. The dihedral angle between the mean planes of the B and E (C19\u2013C24) rings is 87.67\u2005(8)\u00b0.The title mol\u00adecule, (I) C Fig.\u00a01. The di\u00adUncoordW\u22efODhyr and O\u2014HUncoordW\u22efNTrz hydrogen bonds (Table\u00a01Trz\u22efOUncoordW hydrogen bonds (Table\u00a01Cg2\u22efCg1i = 3.5694\u2005(11)\u2005\u00c5 where Cg1 and Cg2 are the centroids of the A and B rings, respectively; symmetry code: (i) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a02; dihedral angle = 2.50\u2005(10)\u00b0] leads to the final three-dimensional structure analysis , Fig.\u00a06c,, Fig.\u00a06d, have a symmetrical distribution with the edges at de + di = 1.85\u2005\u00c5. The H\u22efN/N\u22efN contacts, contributing 7.4% to the overall crystal packing, are shown in Fig.\u00a06e as widely scattered points with the tips at de + di = 2.56\u2005\u00c5. The C\u22efC contacts, Fig.\u00a06f, have an arrow-shaped distribution of points with the tip at de = di = 1.77\u2005\u00c5. Finally, the C\u22efN/N\u22efC inter\u00adactions (2.2%) are reflected in Fig.\u00a06g as tiny characteristic wings with the tips at de + di = 3.44\u2005\u00c5.The overall two-dimensional fingerprint plot, Fig.\u00a06n Table\u00a02 is H\u22efH, c,Table\u00a02 with tridnorm plotted onto the surface are shown for the H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH and H\u22efN/N\u22efH inter\u00adactions in Fig.\u00a07a\u2013d, respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing using standard B3LYP functional and 6\u2013311\u2005G basis-set calculations , hardness (\u03b7), potential (\u03bc), electrophilicity (\u03c9) and softness (\u03c3) are recorded in Table\u00a04H-1,2,3-triazol-4-yl)meth\u00adyl]-2,3-di\u00adhydro-1H-1,3-benzo\u00addiazol-2-one hydrate ring. The energy band gap [\u0394E = ELUMO - EHOMO] of the mol\u00adecule is 5.3468\u2005eV, and the frontier mol\u00adecular orbital energies, EHOMO and ELUMO are \u22126.1633 and \u22120.8166\u2005eV, respectively.The optimized structure of the title compound, (I)To a mixture of 3-methyl-1-(prop-2-yn\u00adyl)-3,4-di\u00adhydro\u00adquinoxalin-2(1H)-one (0.65\u2005mmol) in ethanol (20\u2005ml) was added 1-(azido\u00admeth\u00adyl)benzene (1.04\u2005mmol). The mixture was stirred under reflux for 24\u2005h. After completion of the reaction (monitored by TLC), the solution was concentrated and the residue obtained was purified by column chromatography on silica gel by using as eluent a mixture (hexa\u00adne/ethyl acetate: 9/1). The isolated solid product was recrystallized from ethanol to afford yellow crystals (yield: in 19%).Uiso(H) = 1.2Ueq(C).The experimental details including the crystal data, data collection and refinement are summarized in Table\u00a0510.1107/S2056989019016876/lh5940sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019016876/lh5940Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019016876/lh5940Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019016876/lh5940Isup4.cmlSupporting information file. DOI: 1972575CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III complex compound with pentetic acid is reported. The complex mol\u00adecule is a zwitterion and the GaIII centre is bound in a slightly distorted octa\u00adhedral coordination sphere by two amine N atoms, three carboxyl\u00adate O atoms and one water O atom.The structure of a Ga III complex compound with pentetic acid, [Ga(C14H20N3O10)(H2O)]\u00b73H2O, the GaIII centre is bound in a slightly distorted octa\u00adhedral coordination sphere by two amine N atoms, three carboxyl\u00adate O atoms and one water O atom. The complex mol\u00adecule exists as a zwitterion. In the crystal, the complexes are linked to each other via O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming layers parallel to (001). Three uncoordinating water mol\u00adecules link the complex layers via O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming a three-dimensional network.In the title Ga A complex is easily formed between gallium and DTPA and it has a stability constant of 1023.32, which makes the complex stable against exchange with transferrin \u2005\u00c5]. The C\u2014O bond lengths coordinating to the GaIII atom vary little, with the shortest and longest bonds differing by only 0.019\u2005\u00c5 . The three trans angles, N1\u2014Ga1\u2014O1W, O1\u2014Ga1\u2014O5 and O3\u2014Ga1\u2014N2, are 174.57\u2005(16), 174.05\u2005(12) and 164.97\u2005(13)\u00b0, respectively. The O\u2014Ga\u2014O, O\u2014Ga\u2014N and N\u2014Ga\u2014N bite angles in the chelate rings deviate somewhat from 90\u00b0, ranging from 81.75\u2005(12) to 95.91\u2005(12)\u00b0.The complex mol\u00adecule (abbreviated as Ga-DTPA) is a zwitterion and has a slightly distorted octa\u00adhedral coordination geometry with one water and one amine in the axial positions, and three carboxyl\u00adate groups and one amine in the equatorial positions. The complex consists of three five-membered Ga/N/C/C/O chelate rings and one five-membered Ga/N/C/C/N chelate ring. The Ga\u2014N bonds [Ga1\u2014N1 = 2.081\u2005(4)\u2005\u00c5 and Ga1\u2014N2 = 2.156\u2005(3)\u2005\u00c5] are significantly longer than the Ga\u2014O bonds [Ga1\u2014O1 = 1.933\u2005(3)\u2005\u00c5, Ga1\u2014O3 = 1.925\u2005(3)\u2005\u00c5, Ga1\u2014O5 = 1.964\u2005(3)\u2005\u00c5 and Ga1\u2014O1a and b axes provided in Figs. 2via O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming a three-dimensional network.Packing depictions viewed along the et al., 2016In our survey of the Cambridge Structural Database in acetate buffer (2\u2005mL) adjusted to pH = 4.2 was heated with stirring for dissolution. Gallium nitrate (39.9\u2005mg) was then added to the DTPA solution and the mixture was stirred for at least 10\u2005min at 353\u2005K. The solution was concentrated under ambient pressure at room temperature. When almost all of the solvent had evaporated, methanol was added dropwise to precipitate Ga-DTPA. The precipitate was collected on a 0.22\u2005\u00b5m polyamide filter and dried at room temperature. The obtained Ga-DTPA (1.30\u2005mg) was re-dissolved in ultra-pure water (1\u2005mL) and single crystals suitable for X-ray diffraction were obtained after four weeks by slow diffusion of tetra\u00adhydro\u00adfuran into the aqueous solution, as illustrated in Fig.\u00a04Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018009428/is5497sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018009428/is5497Isup2.hklStructure factors: contains datablock(s) I. DOI: 1852608CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "T was isolated from laboratory tap water during a water quality assessment in University of Malaya, Malaysia. The strain was a Gram\u2010negative, rod\u2010shaped, nonmotile, and aerobic bacterium. Complete genome of P08T comprised of a 2,820,660\u00a0bp chromosome with a G\u00a0+\u00a0C content of 36.43%. Both 16S rRNA phylogeny and phylogenetic tree inferred from the core gene matrix demonstrated that P08T formed a hitherto unknown subline within the family Neisseriaceae. Ortho average nucleotide identity (OrthoANI) values and the percentage of conserved proteins (POCP) calculated from complete genome sequence indicated low relatedness between P08T and its phylogenetic neighbors. Respiratory quinone analysis revealed Q\u20108 as the only detectable quinone. The predominant cellular fatty acids were identified as C14:0, iso\u2010C15:0, and summed feature 3 (C16:1\u03c97c/C16:1\u03c96c). The polar lipids consisted of uncharacterized aminolipid, phosphatidylglycerol, and phosphatidylethanolamine. All aspects of phenotypic and phylogenetic data suggested that strain P08T represents a novel genus within family Neisseriaceae, for which the name Aquella gen. nov. is proposed. The type species of the genus is Aquella oligotrophica sp. nov., and the type strain is P08T (=LMG 29629T =DSM 100970T).A bacterial strain designated as P08 After 3\u00a0days of incubation on R2A agar (BD Difco) at 37\u00b0C, bacteria isolation and purification were performed. The purified strain was routinely cultivated in R2A liquid medium, unless specified. Cells were preserved in 20% (v/v) glycerol at \u221280\u00b0C. The strain P08T has been deposited in the German Collection of Microorganisms and Cell Cultures and the Belgium Coordinated Collections of Microorganisms .Strain P082.2T was extracted using the MasterPureTM DNA purification kit following the manufacturer's protocol. Extracted genomic DNA was sheared and constructed into a template library according to the \u201cGuidelines for Preparing 20\u00a0kb SMRTbell\u2122 Templates.\u201d Genome sequencing was performed in 1 SMRT cell using the PacBio RS II single\u2010molecule real\u2010time (SMRT) sequencing technology . The reads were de novo assembled using the hierarchical genome assembly process (HGAP) algorithm version 2 version 2.10 analysis, the homologous clusters were determined using panX pan\u2010genome pipeline , Luria\u2010Bertani agar (Merck), Pseudomonas agar (BD Difco), MacConkey agar (Merck), R2A agar (BD Difco), and trypticase soy agar (Merck), and the ability to grow in this media was recorded after incubation of 3\u00a0days at 37\u2009\u00b0C. Oxidase and catalase activities were examined with solutions of oxidase reagent (bioM\u00e9rieux) and 3% (v/v) hydrogen peroxide, respectively. The morphology of bacterial cells was observed using tabletop scanning electron microscopy . Cellular motility was tested by the hanging drop method , 0.1\u00a0M NaHCO3/0.1\u00a0M Na2CO3 (pH\u20099.0\u201310.0), and 0.05\u00a0M Na2HPO4/0.1\u00a0M NaOH. Growth on medium added with sodium chloride (NaCl) was determined in R2A medium supplemented with 0, 0.5, 1%\u20136% (w/v) NaCl after 14\u00a0days of incubation at 37\u2009\u00b0C. Growth under anaerobic condition was determined by incubating strain P08T on R2A agar in the Oxoid AnaeroGen system.Growth of strain P08T was tested on R2A agar supplemented with casein , starch , cellulose , urease, Tweens 20, 40, 60, and 80 for hydrolysis activities, respectively on R2A agar (BD Difco) plates. The disks (Oxoid) contained the following antibiotics: ampicillin (10\u00a0\u03bcg), ampicillin/sulbactam (20\u00a0\u03bcg), chloramphenicol (30\u00a0\u03bcg), gentamicin (10\u00a0\u03bcg), kanamycin (30\u00a0\u03bcg), nalidixic acid (30\u00a0\u03bcg), rifampicin (5\u00a0\u03bcg), penicillin G (10\u00a0\u03bcg), streptomycin (10\u00a0\u03bcg), sulfamethoxazole (23.75\u00a0\u03bcg) plus trimethoprim (1.25\u00a0\u03bcg), and tetracycline (30\u00a0\u03bcg). The effect of antibiotics on cell growth was assessed after incubation for 2\u00a0days at 37\u00b0C. The diameter of the antibiotic disks was 6\u00a0mm. The strain was considered susceptible when the diameter of the inhibition zone was>13\u00a0mm, intermediate at 10\u201312\u00a0mm, and resistant at <10\u00a0mm as described by Nokhal and Schlegel , and lesser to all validly described type strains, for instance Neisseria iguana NVSL 85737T (89.4%), Neisseria flavescens ATCC 13120T (89.3%), Paludibacterium paludis KBP\u201021T (89.3%), Uruburuella testudines 07_OD624T (89.3%), and Morococcus cerebrosus CIP 81.93T (89.3%) based on EzBioCloud similarity\u2010based search phylogenetic tree based on 16S rRNA gene sequences. Strain P08T formed a branch clearly separated from the remaining genera from Neisseriaceae. As indicated in Figure T appeared to be more closely related to Neisseriaceae, compared to the type species of genus Prolinoborus, which have a questionable taxonomic status in Neisseriaceae and JGI portal . Out of a total of 2,625 genes in the genome, 2,564 protein\u2010coding gene, 12 rRNAs, and 46 tRNAs were predicted from the chromosome by PGAP analysis designation algorithm version 2 . Intriguingly, strain P08T has the lowest G\u00a0+\u00a0C content among these genomes. The range of G\u00a0+\u00a0C contents ranged widely from the lowest 36.43% in strain P08T to the highest 68.30% in genome of Crenobacter luteus CN10 , iso\u2010C15:0 (36.2%), C16:0 (9.9%), summed feature 3 (C16:1\u03c97c/C16:1\u03c96c) (16.4%), and summed feature 8 (C18:1\u03c97c) (6.8%). Strain P08T exhibited a polar lipid profile consisting of uncharacterized aminolipid, phosphatidylglycerol, and phosphatidylethanolamine value suggested by Ludwig et al. .14:0, iso\u2010C15:0, and summed feature 3 (C16:1\u03c97c/C16:1\u03c96c). The main polar lipids consist of uncharacterized aminolipid, phosphatidylglycerol, and phosphatidylethanolamine. The G\u00a0+\u00a0C content of the DNA of the type strain of the type species is 36.43\u00a0mol%. Based on 16S rRNA sequence analyses, P08T belongs to the Betaproteobacteria. The type species is Aquella oligotrophica P08T.Cells are Gram\u2010negative, aerobic, nonmotile, rod\u2010shaped. Cells are oxidase and catalase negative. Q\u20108 is the only quinone type. Major fatty acids (>10%) are C4.2Aquella oligotrophica .D\u2010galactopyranoside, 2\u2010nitrophenyl\u2010\u03b2\u2010D\u2010galactopyranoside, and sodium pyruvate. Negative reactions toward potassium nitrate, L\u2010tryptophane, D\u2010glucose, L\u2010arginine, urea, esculin, ferric citrate, gelatin (bovine origin), D\u2010glucose, L\u2010arabinose, D\u2010mannose, D\u2010mannitol, N\u2010acetyl\u2010glucosamine, D\u2010maltose, potassium gluconate, capric acid, malic acid, trisodium citrate, phenylacetic acid, L\u2010lysine, L\u2010ornithine, sodium thiosulfate, inositol, D\u2010sorbitol, L\u2010rhamnose, D\u2010sucrose, D\u2010melibiose, and amygdalin. Based on testing with API ZYM kit, C4 esterase, C8 esterase lipase, leucine arylamidase, acid phosphatase, naphthol\u2010AS\u2010BI\u2010phosphohydrolase, \u03b2\u2010galactosidase, and N\u2010acetyl\u2010\u03b2\u2010glucosaminidase activities are detected but negative for activities of C14 lipase, valine arylamidase, trypsin, \u03b1\u2010chymotrypsin, \u03b2\u2010glucuronidase, \u03b1\u2010glucosidase, \u03b2\u2010glucosidase, \u03b1\u2010mannosidase, and \u03b1\u2010fucosidase. Weak enzymatic activities of alkaline phosphatase, cystine arylamidase, and \u03b1\u2010galactosidase are detected. The major fatty acids (> 5%) are C14:0, iso\u2010C15:0, C16:0, summed feature 3 (C16:1\u03c97c/C16:1\u03c96c), and summed feature 8 (C18:1\u03c97c). The following compounds are utilized as sole carbon sources in the GEN III microplate: D\u2010galactose, D\u2010glucuronic acid, D\u2010mannose, glucuronamide, glycyl\u2010L\u2010proline, inosine, L\u2010alanine, L\u2010arginine, L\u2010aspartic acid, L\u2010glutamic acid, L\u2010serine, methyl pyruvate, N\u2010acetyl\u2010D\u2010glucosamine, \u03b1\u2010D\u2010glucose, and \u03b2\u2010hydroxy\u2010D,L\u2010butyric acid. All other substrates in the GEN III microplate are not utilized. Sensitive to ampicillin/sulbactam (20\u00a0\u03bcg), chloramphenicol (30\u00a0\u03bcg), nalidixic acid (30\u00a0\u03bcg), rifampicin (5\u00a0\u03bcg), sulfamethoxazole (23.75\u00a0\u03bcg) plus trimethoprim (1.25\u00a0\u03bcg), and tetracycline (30\u00a0\u03bcg).Exhibits the following properties in addition to those given in the genus description. Cell sizes range from 0.7\u20131.4\u00a0\u03bcm in length and 0.3\u20130.5\u00a0\u03bcm in width was isolated from laboratory tap water collected at University of Malaya, Malaysia.The type strain The authors declare no conflict of interest.KGC supervised the project. LSL carried out the experiments. WSST and LSL wrote the manuscript with support from KGC, KOC, AP, KMG, KWH, and WFY. WSST and LSL analyzed the data.None required.\u00a0Click here for additional data file."} +{"text": "III complex with a Schiff base ligand formed in situ from cyste\u00adamine (2-amino\u00adethane\u00adthiol) and 5-bromo\u00adsalicyl\u00adaldehyde is reported.The crystal structure of novel binuclear Co III complex, [Co2(C9H8BrNOS)2(C18H16Br2N2O2S2)]\u00b7C3H7NO, with a Schiff base ligand formed in situ from cyste\u00adamine (2-amino\u00adethane\u00adthiol) and 5-bromo\u00adsalicyl\u00adaldehyde crystallizes in the space group P21. It was found that during the synthesis the ligand undergoes spontaneous oxidation, forming the new ligand H2L\u2032 having an S\u2014S bond. Thus, the asymmetric unit consists of one Co2(L)2(L\u2032) mol\u00adecule and one DMF solvent mol\u00adecule. Each CoIII ion has a slightly distorted octa\u00adhedral S2N2O2 coordination geometry. In the crystal, the components are linked into a three-dimensional network by several S\u22ef Br, C\u22ef Br, C\u2014H\u22efBr, short S\u22efC contacts as well by weak C\u2014H\u22efO hydrogen bonds. The crystal studied was refined as an inversion twin.The title binuclear Co The synthesis, crystal structure and spectroscopic characterization are described herein.Schiff bases represent one of the most widely used organic compounds. The ability to construct novel ligand systems by means of condensation of a variety of readily available aldehydes and amine makes them and their metal complexes ideal candidates for the construction of novel polynuclear compounds as well for investigation of a large range of properties \u00b0 between the mean planes of atoms O3/N3/C19/C24/C25 and O4/N4/C28/C33/C34 around Co1, and 64.78\u2005(2)\u00b0 between the mean planes of atoms O2/N2/C15/C10/C16 and O1/N1/C1/C6/C7 around Co2. During the synthesis, the ligand is partially oxidized with the formation of a \u2013(CH2)2\u2013S\u2013S\u2013(CH2)2\u2013 bridge. Thus, in contrast to a closely related complex \u2005\u00c5. The Co\u2014S distances in the title complex are in the range 2.207\u2005(3)\u20132.289\u2005(3)\u2005\u00c5, which is generally comparable to the range 2.23\u20132.26\u2005\u00c5 observed for other thio\u00adether\u2013CoIII complexes published earlier [3.596\u2005(2)\u2005\u00c5] and S\u22ef Br [3.364\u2005(2)\u2005\u00c5] contacts, which connect neighboring structural units into chains along [001] are essentially shorter than the sum of the van der Waals radii for the atoms involved [S4\u22efC16 = 3.198\u2005(8)\u2005\u00c5] \u00adethane\u00adthiol\u00adato]-N-(3-thia\u00adpent-5-en\u00adyl)sali\u00adcylaldiminato-N,O)dicobalt(III) aceto\u00adnitrile solvate and [1,8-bis\u00ad-3,6-di\u00adthia\u00adocta\u00adne)cobalt(III) perchlor\u00adate with a di\u00adsulfide moiety . Analysis calculated for C39H39Br4Co2N5O5S4 (M = 1223.49): C,38.28; N, 5.72; H, 3.21%. Found: C, 38.31; N, 5.79; H, 3.28%. The compound is sparingly soluble in CH3CN and good in DMSO, DMF.A solution of KOH in a minimum amount of methanol was added to a solution of 2-amino\u00adethanthiol hydro\u00adchloride in methanol (5\u2005ml) and stirred in an ice bath for 10\u2005min. The white precipitate of solid KCl was removed by filtration and 5-bromo\u00adsalicyl\u00adaldehyde in di\u00admethyl\u00adformamide (10\u2005ml) were added to the filtrate and stirred on air magnetically for 40\u2005min. Cobalt acetate was added to the yellowish solution of the Schiff base formed \u22121 range shows the characteristic azomethine group (\u2013H\u2014C=N) peak at 1616\u2005cm\u22121, indicating the formation of the Schiff base. There are no bands assignable to \u03c5(O\u2014H), indicating the loss of the phenolic hydrogen of the free ligand. In addition, all the characteristic functional group peaks are present in the spectrum. Thus, signals in the 3000\u20133100\u2005cm\u22121 and 1600\u20131400\u2005cm\u22121 regions were assigned to the aromatic C\u2014H and C\u2014C stretches, and weak bands at 544\u2005cm\u22121 and 684\u2005cm\u22121 to the S\u2014S and C\u2014S stretches, respectively. The very strong bands at 1454\u2005cm\u22121 can be attributed to overlapped C\u2014H bending (scissoring) (in the CH3 groups of the solvent mol\u00adecule) and aromatic \u2013C=C stretching vibrations. Another strong band at 1310\u2005cm\u22121 can be assigned to C\u2014O vibrations.The IR spectrum of the title complex in the 4000\u2013400\u2005cm1H NMR spectra, obtained in DMSO-d6 at room temperature using TMS as the inter\u00adnal standard. It revealed an azomethine proton singlet at 8.099 ppm as well the increase in spectroscopic complexity in both the aromatic and aliphatic regions. 1H NMR, DMSO-d6, \u03b4 in ppm: \u2013CH=N, 8.099 (s); aromatic protons (C6H3): 7.94\u20136.52; aliphatic protons (\u2013SCH2CH2N=): 4.44 (m); solvent CH3: 2.96 (s), 2.8 (s). Unfortunately, it could not provide any indication of the dinuclear binding mode, which was revealed only by the X-ray structure determination.The structural assignment of the title compound was supplemented by its Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atom. The crystal studied was refined as an inversion twin with the ratio of the twin components refining to 0.436\u2005(12):0.564\u2005(12).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019007217/lh5903sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019007217/lh5903Isup2.hklStructure factors: contains datablock(s) I. DOI: 1916953CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound is built up from the benzodiazepine ring system linked to the pyridyl and pendant di\u00adhydro\u00adpyran rings. In the crystal, N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds link the mol\u00adecules into a three-dimensional network. A weak C\u2014H \u22ef \u03c0 inter\u00adaction is also observed. 17H18N2O3, is constructed from a benzodiazepine ring system linked to a pendant di\u00adhydro\u00adpyran ring, where the benzene and pendant di\u00adhydro\u00adpyran rings are oriented at a dihedral angle of 15.14\u2005(4)\u00b0. Intra\u00admolecular N\u2014HDiazp\u22efODhydp and C\u2014HDiazp\u22efODhydp (Diazp = diazepine and Dhydp = di\u00adhydro\u00adpyran) hydrogen bonds link the seven-membered diazepine ring to the pendant di\u00adhydro\u00adpyran ring, enclosing S(6) ring motifs. In the crystal, N\u2014HDiazp\u22efODhydp hydrogen bonds link the mol\u00adecules into infinite chains along [10via C\u2014HBnz\u22efODhydp, C\u2014HDhydp\u22efODhydp and C\u2014HMth\u22efODhydp (Bnz = benzene and Mth = meth\u00adyl) hydrogen bonds, forming a three-dimensional network. The observed weak C\u2014HDiazp \u22ef \u03c0 inter\u00adaction may further stabilize the structure. Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (51.1%), H\u22efC/C\u22efH (25.3%) and H\u22efO/O\u22efH (20.3%) inter\u00adactions. Hydrogen bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing.The title compound, C Many members of this family are widely used as anti\u00adconvulsant, anti-anxiety, anti-seizure, analgesic, sedative, anti\u00addepressive and hypnotic or anti-inflammatory agents \u2005\u00c5 (for atom C10), and is oriented at a dihedral angle of 15.14\u2005(4)\u00b0 with respect to the benzene (B: C4\u2013C9) ring. A puckering analysis of the seven-membered diazepine ring (A: N1/N2/C1\u2013C4/C9) gave the parameters QT = 0.6874\u2005(14), q2 = 0.5903\u2005(14), q3 = 0.3523\u2005(14)\u2005\u00c5, \u03c62 = 352.88\u2005(14), \u03c63 = 245.8\u2005(2)\u00b0. In ring A, the N1\u2014C1\u2014C2 [117.10\u2005(2)\u00b0], C1\u2014C2\u2014C3 [114.17\u2005(11)\u00b0], C3\u2014N2\u2014C4 [128.91\u2005(11)\u00b0], N2\u2014C4\u2014C9 [127.38\u2005(12)\u00b0], C4\u2014C9\u2014N1 [126.21\u2005(12)\u00b0] and C9\u2014N1\u2014C1 [130.71\u2005(12)\u00b0] bond angles are enlarged, while the C2\u2014C3\u2014N2 [108.97\u2005(11)\u00b0] bond angle is narrowed, when compared with the corresponding values in the seven-membered diazepine ring in the closely related compound, 3,4-di\u00adhydro-2--4-(4-pyridin-4-yl)-1,5-benzo- diazepine, (II), where the pendant di\u00adhydro\u00adpyran ring is not planar The title compound, (I)4) Fig.\u00a01. Ring C Diazp\u22efODhydp and C\u2014HDiazp\u22efODhydp (Diazp = diazepine and Dhydp = di\u00adhydro\u00adpyran) hydrogen bonds ring motifs s Table\u00a01 link thefs Fig.\u00a01.Diazp\u22efODhydp hydrogen bonds hydrogen bonds analysis and H\u22efN/N\u22efH contacts in the structure contribute only 1.6 and 1.1%, respectively, to the HS. The Hirshfeld surface representations with the function dnorm plotted onto the surface are shown for the H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions in Fig.\u00a07a\u2013c, respectively.The overall two-dimensional fingerprint plot, Fig.\u00a06s Table\u00a02. In the s Table\u00a02. There is Table\u00a01 as well s Table\u00a01 and is s\u22efO Fig.\u00a06e and H\u22ef\u22efH Fig.\u00a06f contacet al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing in ethanol (40\u2005ml) was refluxed for 1\u2005h. After cooling to room temperature, the colourless inter\u00admediate solid compound, a mono-Schiff base, was obtained in 70% yield. The inter\u00admediate was refluxed in acetone (10\u2005ml) for 1h. After cooling, the crystals formed were filtered and dried (yield: 65%).A solution of de\u00adhydro\u00adacetic acid and Uiso(H) = kUeq, where k = 1.5 for methyl H atoms and 1.2 for the other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019000689/lh5890sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019000689/lh5890Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019000689/lh5890Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019000689/lh5890Isup4.cmlSupporting information file. DOI: 1890950CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structures of two new forms of cobalt\u2013pyridine\u2013sulfate complexes are presented. The feature infinite chains of metal\u2013pyridine units connected by bridging sulfate anions, which are distinct from the only previously reported structure of a cobalt\u2013pyridine\u2013sulfate compound. catena-poly[[tetra\u00adkis\u00ad(pyridine-\u03baN)cobalt(II)]-\u03bc-sulfato-\u03ba2O:O\u2032], [Co(SO4)(C5H5N)4]n, (1), and catena-poly[[tetra\u00adkis\u00ad(pyridine-\u03baN)cobalt(II)]-\u03bc-sulfato-\u03ba3O:O\u2032,O\u2032\u2032-[bis\u00ad(pyridine-\u03baN)cobalt(II)]-\u03bc-sulfato-\u03ba3O,O\u2032:O\u2032\u2032]n, [Co2(SO4)2(C5H5N)6]n, (2), are reported. Compound (1) displays a polymeric structure, with infinite chains of CoII cations adopting octa\u00adhedral N4O2 coordination environments that involve four pyridine ligands and two bridging sulfate ions. Compound (2) is also polymeric with infinite chains of CoII cations. The first Co center has an octa\u00adhedral N4O2 coordination environment that involves four pyridine ligands and two bridging sulfate ligands. The second Co center has an octa\u00adhedral N2O4 coordination environment that involves two pyridine ligands and two bridging sulfate ions that chelate the Co atom. The structure of (2) was refined as a two-component inversion twin.The solid-state structures of two cobalt\u2013pyridine\u2013sulfate compounds, namely When grown out, the cobalt ion shows an octa\u00adhedral coordination environment . The equatorial positions of the octa\u00adhedron are occupied by four pyridine ligands in a square-planar arrangement. The CoN4 unit exhibits planarity enforced by symmetry, with cis N\u2014Co\u2014N angles of 86.45\u2005(6) and 93.55\u2005(6)\u00b0. To complete the octa\u00adhedron, the axial positions are occupied by two sulfate ions, with an inversion enforced O\u2014Co\u2014O angle of 180\u00b0 and cis O\u2014Co\u2014N angles of 88.87\u2005(6) and 91.67\u2005(6)\u00b0. The pyridine rings are rotated from the CoN4 plane by dihedral angles of 47.30\u2005(10) and 78.33\u2005(9)\u00b0. The 78.33\u2005(9)\u00b0 angles are constrained by two C\u2014H\u22efO inter\u00adactions between the ortho hydrogen atoms and the two trans sulfates consists of two cobalt atoms, six coordinated pyridines and two sulfate anions . There are two crystallographically unique cobalt atoms, with Co1 displaying an octa\u00adhedral N4O2 coordination environment and Co2 exhibiting an octa\u00adhedral N2O4 coordination geometry.The asymmetric unit of the purple crystals of to 93.18\u2005(12)\u00b0, and the trans O\u2014Co\u2014O angle is 173.43\u2005(12)\u00b0. The planes of the four pyridine rings are rotated from the equatorial CoN4 plane by dihedral angles of 58.6\u2005(2), 64.6\u2005(2), 65.6\u2005(2), and 73.1\u2005(2)\u00b0. Two of the rings show one C\u2014H\u22efO inter\u00adaction with an ortho hydrogen atom, one ring shows two C\u2014H\u22efO inter\u00adactions with two ortho hydrogen atoms, and the fourth ring shows no C\u2014H\u22efO inter\u00adactions \u00b0. The two sulfate ligands exhibit O\u2014Co\u2014O bite angles of 65.90\u2005(10) and 66.37\u2005(10)\u00b0. The other cis O\u2014Co\u2014O angles are 86.87\u2005(11), 98.98\u2005(11), and 102.84\u2005(11)\u00b0, and the six cis N\u2014Co\u2014O angles range from 92.49\u2005(12) to 98.33\u2005(13)\u00b0. Each pyridine ring is involved in ortho C\u2014H\u22efO inter\u00adactions are linked together into infinite chains along the [001] direction through sulfate anions with O\u2014S\u2014O bridges . Between each successive tetra\u00adpyridine cobalt unit, there are parallel slipped \u03c0\u2013\u03c0 inter\u00adactions .The Cos Figs.\u00a03a, 4 \u25b8aII atoms in compound (2) are linked together into infinite chains along the [111] direction through the sulfate anions . The chain alternates between tetra\u00adpyridine cobalt units and di\u00adpyridine cobalt units. No \u03c0\u2013\u03c0 inter\u00adactions are observed in the crystal.The Cos Figs.\u00a03b, 4 \u25b8b3(SO4)3(C5H5N)11)]n, which was grown at a lower concentration of cobalt. This structure shows two successive octa\u00adhedral cobalt atoms with N4O2 coordination, where each atom is coordinated to four pyridines and two bridging sulfates. The third cobalt atom in the chain shows N3O3 coordination where three pyridines are bound and there are two sulfates bound, one of which is chelating to the cobalt and (2). In compound (1), every cobalt atom possesses an octa\u00adhedral N4O2 coord\u00adin\u00adation. This complex is isostructural with the structure observed for the iron and nickel pyridine\u2013sulfate complexes , the cobalt atoms alternate between N4O2 coordination and N2O4 coordination. This tetra\u00adpyridine/bi\u00adpyridine alternation is similar to what is observed in the zinc\u2013pyridine\u2013sulfate structure, which alternates between octa\u00adhedral and tetra\u00adhedral zinc centers. In the case of cobalt, the bis\u00ad(pyridine) cobalt center is still octa\u00adhedral because the two coordinated sulfates both chelate to the cobalt. The end result is an infinite chain of octa\u00adhedral cobalt atoms, which is true in compound (1) and the previously reported cobalt\u2013pyridine\u2013sulfate complex. The methane\u00adsulfato complexes of cobalt (II) have also been reported as octa\u00adhedral tetra\u00adkis\u00ad(pyridine), [Co(SO3CH3)2(py)4], and octa\u00adhedral bis\u00ad(pyridine), [Co(SO3CH3)2(py)2], compounds, consistent with the two independent cobalt centers observed in (2) , 40\u2005mg of cobalt sulfate hepta\u00adhydrate (J. T. Baker) was dissolved in pyridine and distilled water (100\u2005\u00b5L) in a 20\u2005mL vial. The vial was heated to 338\u2005K for 48\u2005h, after which single crystals suitable for X-ray diffraction studies were isolated from the reaction mixture.For compound (2), 48\u2005mg of cobalt sulfate hepta\u00adhydrate (J. T. Baker) was dissolved in pyridine and distilled water (30\u2005\u00b5L) in a 20\u2005mL vial. The vial was heated to 358\u2005K for 48\u2005h, after which single crystals suitable for X-ray diffraction studies were isolated from the reaction mixture.For compound (SHELXL) by full-matrix least squares on F2. Hydrogen atoms were placed in calculated positions and then refined with a riding model with C\u2014H bond lengths of 0.95\u2005\u00c5 and with isotropic displacement parameters set to 1.20 Ueq of the parent C atom. The structre of (2) was refined as a two-component inversion twin, BASF = 0.165\u2005(13).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901901538X/sj5586sup1.cifCrystal structure: contains datablock(s) 1, 2, I. DOI: 10.1107/S205698901901538X/sj55861sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S205698901901538X/sj55862sup3.hklStructure factors: contains datablock(s) 2. DOI: 1965662, 1965663, 1965662, 1965663CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The structures of the two isomeric hydrogen-bonded 1:1 co-crystals of 3-chloro-2-nitro\u00adbenzoic acid with 5-nitro\u00adquinoline and 6-nitro\u00adquinoline, and the 1:1 salt of 3-chloro-2-nitro\u00adbenzoic acid with 8-hy\u00addroxy\u00adqunoline have been determined at 190\u2005K. In each crystal, the acid and base mol\u00adecules are linked by a short O\u2014H\u22efN or N\u2014H\u22efO hydrogen bond. 7H4ClNO4\u00b7C9H6N2O2, the acid and base mol\u00adecules are held together by O\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds. In compound (III), C9H8NO+\u00b7C7H3ClNO4\u2212, an acid\u2013base inter\u00adaction involving H-atom transfer occurs and the H atom is located at the N site of the base mol\u00adecule. In the crystal of (I), the hydrogen-bonded acid\u2013base units are linked by C\u2014H\u22efO hydrogen bonds, forming a tape structure along the b-axis direction. Adjacent tapes, which are related by a twofold rotation axis, are linked by a third C\u2014H\u22efO hydrogen bond, forming wide ribbons parallel to the (via \u03c0\u2013\u03c0 inter\u00adactions between the quinoline ring systems [centroid\u2013centroid distances = 3.4935\u2005(5)\u20133.7721\u2005(6)\u2005\u00c5], forming layers parallel to the ab plane. In the crystal of (II), the hydrogen-bonded acid\u2013base units are also linked into a tape structure along the b-axis direction via C\u2014H\u22efO hydrogen bonds. Inversion-related tapes are linked by further C\u2014H\u22efO hydrogen bonds to form wide ribbons parallel to the \u20133.9247\u2005(9)\u2005\u00c5], and the mol\u00adecular chains are linked into layers parallel to the ab plane through these inter\u00adactions.The structures of three compounds of 3-chloro-2-nitro\u00adbenzoic acid with 5-nitro\u00adquinoline, (I), 6-nitro\u00adquinoline, (II), and 8-hy\u00addroxy\u00adquinoline, (III), have been determined at 190\u2005K. In each of the two isomeric compounds, (I) and (II), C The \u0394pThe mol\u00adecular structure of (I)The mol\u00adecular structure of (II)+\u2014H\u22efO\u2212 hydrogen bond (Table\u00a03The mol\u00adecular structure of (III)d Table\u00a03. In the i and C14\u2014H14\u22efO5i; symmetry codes as in Table\u00a01b-axis direction. Adjacent tapes, which are related by a twofold rotation axis, are linked by a third C\u2014H\u22efO hydrogen bond (C13\u2014H13\u22efO6ii), forming wide ribbons parallel to the \u2212x\u00a0+\u00a01, y, \u2212z\u00a0+\u00a02].In the crystal of (I)ne Fig.\u00a04. These rne Fig.\u00a05. The cenb-axis direction via C\u2014H\u22efO hydrogen bonds via weak \u03c0\u2013\u03c0 inter\u00adactions \u2212x, \u2212y\u00a0+\u00a02, \u2212z; (v) x\u00a0\u2212\u00a01, y\u00a0+\u00a01, z; (vi) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01]. A pair of short O\u22efN contacts between the nitro groups of the base mol\u00adecule are alsso observed.In the crystal of (II)ne Fig.\u00a06. The acins Fig.\u00a07, so formvia N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds \u2005\u00c5]; Cg1 and Cg3 are, respectively, the centroids of the C1\u2013C6 ring of the anion and the C11\u2013C16 ring of the cation. In addition to the \u03c0\u2013\u03c0 inter\u00adaction (Cg1\u22efCg3i), other \u03c0\u2013\u03c0 inter\u00adactions are observed; the centroid\u2013centroid distances are 3.5469\u2005(6), 3.8550\u2005(6) and 3.5133\u2005(6)\u2005\u00c5, respectively, for Cg1\u22efCg2iii, Cg1\u22efCg3iii and Cg1\u22efCg4iii, where Cg2 and Cg4 are the centroids of the N2/C8\u2013C11/C16 and N2/C8\u2013C16 rings of the cation, respectively is observed between the layers.In the crystal of compound (III)on Fig.\u00a08. In the ns Fig.\u00a09, and theet al., 2016et al., 2002et al., 2018et al. , 2.561\u2005(1), 2.540\u2005(2)\u20132.571\u2005(2), 2.573\u2005(1) and 2.613\u2005(3)\u2005\u00c5, respectively. Furthermore, in the short hydrogen bonds of AJIWOG, CALJUW and NOVLAN, the H atom is disordered over two positions. On the other hand, the compounds (I)Ka values of 0.98, 1.42 and 3.02, respectively, show longer O\u22efN distances of 2.673\u2005(1), 2.631\u2005(1) and 2.636\u2005(1)\u2005\u00c5, which suggests that the \u0394pKa value is not an effective measure of hydrogen-bond strength in the 3-chloro-2-nitro\u00adbenzoic acid\u2013organic base system.A search of the Cambridge Structural Database , in which the O\u22efN distance of the N\u2014H\u22efO hydrogen bond is 2.644\u2005(3)\u2005\u00c5.A search for organic co-crystals/salts of 5-nitro\u00adquinoline showed six structures. Limiting the search to benzoic acid derivatives gave two hits, namely, 3-amino\u00adbenzoic acid\u20135-nitro\u00adquinoline (1/1) \u2013(III), were obtained by slow evaporation from aceto\u00adnitrile solutions of 3-chloro-2-nitro\u00adbenzoic acid with quinoline derivatives in a 1:1 molar ratio at room temperature [100\u2005ml aceto\u00adnitrile solution of 3-chloro-2-nitro\u00adbenzoic acid (0.39\u2005g) and 5-nitro\u00adquinoline (0.34\u2005g) for (I)O = 0.872\u2005(19)\u2005\u00c5 in (III)Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989019012799/lh5922sup1.cifCrystal structure: contains datablock(s) global, I, II, III. DOI: 10.1107/S2056989019012799/lh5922Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019012799/lh5922IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989019012799/lh5922IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 1953605, 1953604, 1953603CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dimeric mol\u00adecules of the complexes crystallize together with water and methanol mol\u00adecules, with which they form a variety of weak and medium-strength hydrogen bonds.Oxalate-bridged, centrosymmetric binuclear complexes of gadolinium(III) and dysprosium(III) with hexa\u00addentate bbpen Ln(bbpen)Cl] and potassium oxalate monohydrate in water/methanol produced the solvated centrosymmetric isostructural binuclear bis\u00ad{dilanthanide(III)}\u2013methanol\u2013water (1/4/4) complexes, [Ln2(C28H28N4O2)2(C2O4)]\u00b74CH3OH\u00b74H2O, with lanthanide(III) = gadolinium(III) (Ln = Gd) and dysprosium(III) (Ln = Dy), in high yields (ca 70%) directly from the reaction mixtures. In both complexes, the lanthanide ion is eight-coordinate and adopts a distorted square-anti\u00adprismatic coordination environment. The triclinic (PPLATON . These two new compounds are of inter\u00adest with respect to magnetic properties.The reaction between mononuclear [Spek 2015. Acta Cr The lanthanide cations assume square-anti\u00adprismatic coordination environments while the d-block metal is octa\u00adhedrally coordinated 2]+ complex had previously been employed to produce binuclear [Dy2(\u03bc-ox)(HBpz3)4]\u00b72CH3CN\u00b7CH2Cl2, this time with oxalate (ox2\u2013) as the bridging ligand. Direct current (DC) magnetic susceptibility measurements performed with this dimeric compound revealed the presence of an intra\u00admolecular ferromagnetic inter\u00adaction between the DyIII cations 3{Ln(bbpen)}3] -N,N\u2032-bis\u00ad(pyridin-2-ylmeth\u00adyl)ethyl\u00adenedi\u00adamine) via modular synthesis employing [Ln(bbpen)Cl] (LnIII = Gd or Dy) and K3[M(ox)3] (MIII = Cr or Co) as building blocks in a 3:1 proportion. The syntheses with gadolinium(III) and chromium(III) produced colourless crystals of the binuclear complex [{Gd(bbpen)}2(\u03bc-ox)]\u00b74CH3OH\u00b74H2O, as revealed by single crystal X-ray diffraction analysis. The formation of this dimer is explained by dissociation of [Cr(ox)3]3\u2013 into {Cr(ox)2(OH2)2}\u2212 and ox2\u2013 in aqueous solution +. Structural elucidation of this otherwise unexpected product prompted us to try and perform its targeted preparation with both gadolinium(III) and dysprosium(III) in good yields.In our research group, we first attempted to prepare heterometallic complexes of general formula [Ln(bbpen)}2(\u03bc-ox)] products [Ln = Gd (1) or Dy (2)], prepared from the direct reaction between [Ln(bbpen)Cl] and K2C2O4\u00b7H2O in water/methanol media.In this paper we report the rational synthesis and the crystal and mol\u00adecular structures of the two binuclear and solvated [{1 and 2 are isostructural and crystallize in the PLn2(\u03bc-ox)(bbpen)2] mol\u00adecules or dysprosium(III) (2) are eight-coordinate; the [Ln(bbpen)]+ units are connected to one another by oxalate bridging in the usual bis\u00ad(bidentate) coordination mode. The ox2\u2013 ligand lies about an inversion centre. The coordination sphere of the lanthanide(III) ion is formed by an N4O2 donor set from the bbpen2\u2013 ligand and two oxygen atoms from the bridging oxalate. In 1 and 2 each metal cation has a distorted square-anti\u00adprismatic coordination environment and 2.24\u2005\u00c5 (2). The non-bonding Dy\u22efDy distance in 2, 6.1488\u2005(17)\u2005\u00c5, is close to the analogous distance of 6.14\u2005\u00c5 in [Dy2(\u03bc-ox)(HBpz3)4]\u00b72CH3CN\u00b7CH2Cl2 \u2212x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z] are also similar to those reported for the dysprosium(III)\u2013hydro\u00adtris(pirazolylborate) dimer mentioned above. The slightly decreased crystal volume of the Dy compound [1626.3\u2005(7)\u2005\u00c53] compared with that of the Gd compound [1633.7\u2005(3)\u2005\u00c53] is a perfect match with the smaller effective ionic radius of eight-coordinate DyIIIversus GdIII product 2 gives rise to very similar results.Compounds es Fig.\u00a01 in whichnt Fig.\u00a02, as indiIn both structures, the hydrogen atoms from the crystallizing solvents (water and methanol) participate in an extensive three-dimensional hydrogen-bonding network that may be described as medium-strength inter\u00admolecular inter\u00adactions Tables 1 and 2 \u25b8.1 and 2. As seen in Fig.\u00a03W\u2014H\u22efO2iii \u2018bridge\u2019, as well as a symmetry-related chain on both sides of the plane formed by the metal and oxalate ions. The water mol\u00adecules in these chains also connect one dimer to another through weak C1\u2014H1B\u22efO1Wii inter\u00adactions mol\u00adecules, half of which refine well and are depicted in Fig.\u00a03ns Fig.\u00a04 and 2 \u25b8.PLATON for O\u22efO30. On the other hand, any possible inter\u00adaction involving the phenolate oxygen atoms would be very weak, with the shortest O\u22efO contact with the disordered solvents being longer than 4.0\u2005\u00c5.The other half of the solvent mol\u00adecules in the unit cell, the electron densities of which have been removed with the SQUEEZE routine in 2\u2013 and related ligands appear in the literature complexes with bbpen al. 2005, and by al. 2000.LnCl3\u00b76H2O (LnIII = Gd or Dy) and K2C2O4\u00b7H2O were purchased from Aldrich and used without purification. N,N\u2032-Bis(2-hy\u00addroxy\u00adbenz\u00adyl)-N,N\u2032-bis\u00ad(pyridin-2-ylmeth\u00adyl)ethyl\u00adene\u00addi\u00adamine (H2bbpen) Cl] precursors, with Ln = Gd or Dy ]\u00b74CH3OHSynthesis of [{Gd(bbpen)}\u00b72O (compound 1)4H2C2O4\u00b7H2O in 1.0\u2005ml of water was slowly added to a methanol solution of 61.1\u2005mg (0.0947\u2005mmol) of [Gd(bbpen)Cl]. The colourless reaction mixture was stirred at room temperature for ca 5\u2005min, and was then cooled down to 277\u2005K to give block-shaped colourless crystals after four days. These were isolated by filtration, washed with diethyl ether and dried. Total yield: 49.0\u2005mg (68.6%) based on the [{Gd(bbpen)}2(\u03bc\u2013ox)]\u00b74CH3OH\u00b74H2O formulation, compound 1. FTIR : 3362, 3198 ; 1655 ; 1590, 1568 , 1290 , 762 and 768 . Product 1 is soluble in aceto\u00adnitrile, 1,2-di\u00admeth\u00adoxy\u00adethane (dme), di\u00adchloro\u00admethane and tetra\u00adhydro\u00adfuran. Elemental analysis: calculated for 1 (C62H80Gd2N8O16) C 49.39, H 5.35, N 7.43%. Found: C 48.56, H 5.49, N 7.45%.A solution of 8.11\u2005mg (0.0440\u2005mmol) of K2(\u03bc\u2013ox)]\u00b74CH3OHSynthesis of \u00b74CH3OH\u00b74H2O formulation, compound 2. FTIR : 3363, 3198 , 1590 ; 1570 (m), 1481 (s), 1459 ; 1290 , 762 and 768 . The product solubility is similar to that described for 1. Elemental analysis: calculated for 2 (C62H80Dy2N8O16) C 49.04, H 5.31, N 7.38%. Found: C 49.02, H 5.71, N 7.56%.A mixture of 61.0\u2005mg (0.0938\u2005mmol) of [Dy(bbpen)Cl] in 9.0\u2005ml of methanol and 8.90\u2005mg (0.0483\u2005mmol) of K1 and 2 showed high susceptibility to the loss of the crystallization solvent mol\u00adecules once removed from the mother liquor. Hydrogen atoms in 1 and 2 were included in idealized positions with methyl, methyl\u00adene and aromatic C\u2014H distances set at 0.98, 0.99 and 0.95\u2005\u00c5, respectively, and O\u2014H at 0.84\u2005\u00c5 and refined as riding with Uiso(H) = 1.2\u20131.5Ueq. Hydrogen atoms on the water mol\u00adecules were located in difference-Fourier maps and were refined with distance restraints .Crystal data, data collection and structure refinement details for the two structures are summarized in Table\u00a03PLATON 1, 2. DOI: 10.1107/S2056989019002998/wm54891sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989019002998/wm54892sup3.hklStructure factors: contains datablock(s) 2. DOI: 1899963, 1899964CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N-oxide and solvent mol\u00adecules are reported. The pyridine N--oxide, 2-methyl\u00adpyridine N-oxide, 3-methyl\u00adpyridine N-oxide, and 4-methyl\u00adpyridine N-oxide complexes all form similar structures with slight differences owing to the substituent group effects.The synthesis and crystal structures of four dimeric complexes composed of manganese(II) dibromide, a pyridine N-oxides, viz. pyridine N-oxide (PNO), 2-methyl\u00adpyridine N-oxide (2MePNO), 3-methyl\u00adpyridine N-oxide (3MePNO), and 4-methyl\u00adpyridine N-oxide (4MePNO). The compounds are bis\u00ad(\u03bc-pyridine N-oxide)bis\u00ad[aqua\u00addibromido\u00ad(pyridine N-oxide)manganese(II)], [Mn2Br4(C5H5NO)4(H2O)2] (I), bis\u00ad(\u03bc-2-methyl\u00adpyridine N-oxide)bis\u00ad[di\u00adaqua\u00addibromido\u00admanganese(II)]\u20132-methyl\u00adpyridine N-oxide (1/2), [Mn2Br4(C6H7NO)2(H2O)4]\u00b72C6H7NO (II), bis\u00ad(\u03bc-3-methyl\u00adpyridine N-oxide)bis\u00ad[aqua\u00addibromido\u00ad(3-methyl\u00adpyridine N-oxide)manganese(II)], [Mn2Br4(C6H7NO)4(H2O)2] (III), and bis\u00ad(\u03bc-4-methyl\u00adpyridine N-oxide)bis\u00ad[di\u00adbromido\u00admethanol(4-methyl\u00adpyridine N-oxide)manganese(II)], [Mn2Br4(C6H7NO)4(CH3OH)2] (IV). All the compounds have one unique MnII atom and form a dimeric complex that contains two MnII atoms related by a crystallographic inversion center. Pseudo-octa\u00adhedral six-coordinate manganese(II) centers are found in all four compounds. All four compounds form dimers of Mn atoms bridged by the oxygen atom of the PNO ligand. Compounds I, II and III exhibit a bound water of solvation, whereas compound IV contains a bound methanol mol\u00adecule of solvation. Compounds I, III and IV exhibit the same arrangement of mol\u00adecules around each manganese atom, ligated by two bromide ions, oxygen atoms of two PNO ligands and one solvent mol\u00adecule, whereas in compound II each manganese atom is ligated by two bromide ions, one O atom of a PNO ligand and two water mol\u00adecules with a second PNO mol\u00adecule inter\u00adacting with the complex via hydrogen bonding through the bound water mol\u00adecules. All of the compounds form extended hydrogen-bonding networks, and compounds I, II, and IV exhibit offset \u03c0-stacking between PNO ligands of neighboring dimers.Four manganese(II) bromide coordination complexes have been prepared with four pyridine N-oxides have inter\u00adesting binding modes that facilitate the growth of unique coordination structures. Their utility to facilitate organic oxotransfer reactions has been well documented over the years and manganese(II) metal ions with a single bifunctional ligand containing an acetate and N-oxide moiety dimeric complexes, using pyridine N-oxide (PNO) and its mono-methyl-substituted forms, 2-methyl\u00adpyridine N-oxide (2MePNO), 3-methyl\u00adpyridine N-oxide (3MePNO), and 4-methyl\u00adpyridine N-oxide (4MePNO). This was done to study the impact of substitution of the pyridine on the two- and three-dimensional solid-state structures, and to compare them to previous structures in which the bromide ions are replaced with chloride ions.Herein, we report the synthesis and solid-state structures of four pyridine General structural detailsN-oxide complexes form dimers consisting of two MnII atoms related by an inversion center; the dimer contains a six-coordinate metal center at each MnII ion with four donor oxygen atoms and two bromides. The Mn1\u22efMn1\u2032 dimer is bound trans by two \u03bc2-1,1-PNO ligands, and the octa\u00adhedral environment is completed by a water mol\u00adecule of hydration or a solvent mol\u00adecule, non-bridging PNO ligands, and bromide ions. The dimer is constructed from symmetry-related atoms and mol\u00adecules using a crystallographic inversion center of the space group (PP21/n). The mol\u00adecular structures of compounds I, II, III and IV are given in Figs. 1The pyridine Specific structural detailsI and 2.235\u2005(2)\u2005\u00c5 for Mn1\u2014O1 and Mn1\u2014O1i, respectively, which is unremarkable for compounds of MnII and pyridine N-oxide \u2005\u00c5. The octa\u00adhedral geometry around the Mn atoms is significantly distorted with the O1\u2014Mn1\u2014O1i bond angle measuring 69.66\u2005(9)\u00b0; the other bond angles are within ca 9\u00b0 of 90\u00b0. These bond angles and bond lengths are similar to those for other MnII halide PNO structures \u2005\u00c5 .Compound I Fig.\u00a01 crystallII and 2.321\u2005(2)\u2005\u00c5 for Mn1\u2014O1 and Mn1\u2014O1i, respectively. The two bound water mol\u00adecules have Mn\u2014O bond lengths of 2.237\u2005(3) and 2.157\u2005(3)\u2005\u00c5 for Mn1\u2014O3 and Mn1\u2014O4, respectively, and are similar to those reported previously \u2005\u00c5. Once again the octa\u00adhedral geometry around the Mn atoms is significantly distorted with the O1\u2014Mn1\u2014O1i bond angle measuring 74.40\u2005(9)\u00b0. The other bond angles are within ca 11\u00b0 of 90\u00b0. The dimer forms an intra\u00admolecular hydrogen bond between O3 and Br1i with a hydrogen-bond distance of 2.44\u2005(2)\u2005\u00c5 . In the asymmetric unit there is a second PNO mol\u00adecule inter\u00adacting with the complex via hydrogen bonding through the bound water mol\u00adecules and 2.219\u2005(3)\u2005\u00c5 for Mn1\u2014O2 and Mn1\u2014O2i, respectively. The non-bridging Mn1\u2014O1 bond is 2.129\u2005(3)\u2005\u00c5, and the bound water Mn1\u2014O3 bond distance is 2.245\u2005(3)\u2005\u00c5. The bound bromide ions have bond distances of Mn1\u2014Br1 = 2.7237\u2005(7)\u2005\u00c5 and Mn1\u2014Br2 = 2.5687\u2005(7)\u2005\u00c5; again the difference in Mn\u2014Br bond distances can be attributed to the hydrogen-bonding inter\u00adactions that exist with Br1 but not with Br2 (Table\u00a03i distance is 3.6497\u2005(13)\u2005\u00c5. The octa\u00adhedral geometry around the Mn atoms is significantly distorted with the O2\u2014Mn1\u2014O2i bond angle measuring 69.05\u2005(11)\u00b0 the other bond angles are within ca 11\u00b0 of 90\u00b0. The dimer forms an intra\u00admolecular hydrogen bond between O3 and Br1ii with a hydrogen-bond distance of 2.55\u2005(2)\u2005\u00c5 .Compound II Fig.\u00a03 crystall2 Table\u00a03. The Mn1IV and 2.230\u2005(3)\u2005\u00c5 for Mn1\u2014O2 and Mn1\u2014O2i, respectively. The non-bridging Mn1\u2014O1 bond is 2.116\u2005(3)\u2005\u00c5, and the bound methanol Mn1\u2014O3 bond distance is 2.225\u2005(3)\u2005\u00c5. The bound bromide ions have bond distances of Mn1\u2014Br1 = 2.7181\u2005(7)\u2005\u00c5 and Mn1\u2014Br2 2.5806\u2005(7)\u2005\u00c5, again the difference in Mn\u2014Br bond distance can be attributed to the hydrogen-bonding inter\u00adactions (Table\u00a04i distance is 3.61254\u2005(12)\u2005\u00c5. The octa\u00adhedral geometry around the Mn atoms is significantly distorted with the O2\u2014Mn1\u2014O2i bond angle measuring 70.77\u2005(11)\u00b0 the other bond angles are within 13\u00b0 of 90\u00b0. The dimer forms an intra\u00admolecular hydrogen bond between O3 and Br1i with a hydrogen-bond distance of 2.41\u2005(2)\u2005\u00c5 .Compound IV Fig.\u00a04 crystalls Table\u00a04. The Mn1I, the dimers are linked by Owater\u2014H\u22efBr hydrogen bonds, forming chains parallel to the [100] direction; see Table\u00a01ac plane \u2005\u00c5 and O4\u22efO2ii = 2.721\u2005(4)\u2005\u00c5 (Table\u00a02B\u22efBr1i with a distance of 2.44\u2005(2)\u2005\u00c5 , with an inter-centroid distance of the stacked aromatic rings of 3.516\u2005(4)\u2005\u00c5, so forming layers parallel to the ac plane \u2005\u00c5, with a significant centroid shift of 3.221\u2005(9)\u2005\u00c5 preventing \u03c0-stacking. Neighboring dimers are linked by O\u2014H\u22efBr hydrogen-bonds forming chains parallel to the a axis. There are two observed inter\u00adactions, O3\u2014H3A\u22efBr1i with a distance of 2.60\u2005(2)\u2005\u00c5 and O3\u2014H3B\u22efBr1ii with a distance of 2.55\u2005(2)\u2005\u00c5 . Five of these structures contain derivatives of pyridine N-oxides and one of them is a 4,4\u2032-dipyridal N,N\u2032-dioxide (H2O)]n, [MnCl2(2MPNO)(H2O)]n, and [MnCl2(3MPNO)(H2O)2]2 2(H2O)6I2]I2 bromide tetra\u00adhydrate was dissolved in a minimal amount (20\u2005ml) of methanol. Two molar equivalents of pyridine N-oxide were also dissolved in methanol. The solutions were mixed and stirred for 10\u2005min and the solvent was allowed to evaporate to produce X-ray quality crystals . Selected IR bands 3470 , 1471 (s), 1216 (s), 833 (s) 773 (m), 669 (m), 558 (m). Analysis calculated for C20H24N4Mn2Br4O6: C, 28.40; H, 2.86; N, 6.62%. Found: C, 28.13; H, 2.86; N, 6.50%.Compound II: Manganese(II) bromide tetra\u00adhydrate was dissolved in a minimal amount (20\u2005ml) of methanol. Two molar equivalents of 2-methyl\u00adpyridine N-oxide were also dissolved in methanol. The solutions were mixed and stirred for 10\u2005min and the solvent was allowed to evaporate to produce X-ray quality crystals . Selected IR bands 3349 , 1600 (m), 1461 (s), 1195 (s) 842 (m), 772 (s), 557 (m). Analysis calculated for C24H36N4Mn2Br4O8: C, 30.73; H, 3.87; N, 5.97%. Found: C, 30.30; H, 3.62; N, 6.17%.Compound III: Manganese(II) bromide tetra\u00adhydrate was dissolved in a minimal amount (20\u2005ml) of methanol. Two molar equivalents of 3-methyl\u00adpyridine N-oxide were also dissolved in methanol. The solutions were mixed and stirred for 10\u2005min and the solvent was allowed to evaporate to produce a powder . X-ray quality crystals were grown by recrystallizing a second time by slow evaporation from methanol. Selected IR bands 3373 , 1631 (s), 1492 (m), 1260 (m), 1163(s), 943 (m), 802 (m).Compound IV: Manganese(II) bromide tetra\u00adhydrate was dissolved in a minimal amount (20\u2005ml) of methanol. Two molar equivalents of 4-methyl\u00adpyridine N-oxide were also dissolved in methanol. The solutions were mixed and stirred for 10\u2005min and the solvent was allowed to evaporate to produce a powder . X-ray quality crystals were grown by recrystallizing a second time from methanol with a slower evaporation rate. Selected IR bands 3227 , 3004 (m), 1670 (m), 1494(s), 1213 (s), 852(s), 763(s).I and II have been reported analytically pure, whereas III and IV were not isolated analytically pure. The FT\u2013IR spectra of the four N-oxide complexes all exhibit broad absorbances in the 3500\u20133100\u2005cm\u22121 region characteristic of the \u03bd(O\u2014H) of the coordinated water or methanol mol\u00adecules. In addition, the \u03bd(N\u2014O) stretching frequency that is due to the N-oxide pyridyl moiety is observed in the region between 1260 and 1195\u2005cm\u22121, as noted previously \u2005\u00c5 and refined with Uiso(H) = 1.5Ueq(O). In compound IV, the hydroxyl H atom was located in a difference-Fourier map and refined with O\u2014H distance restrained to 0.85\u2005(1)\u2005\u00c5 and with Uiso(H) = 1.5Ueq(O). All carbon-bound H atoms were positioned geometrically and refined as riding: C\u2014H = 0.95\u20130.98\u2005\u00c5 with Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989019010557/su5505sup1.cifCrystal structure: contains datablock(s) Global, II, III, IV, I. DOI: 10.1107/S2056989019010557/su5505Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019010557/su5505IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989019010557/su5505IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 10.1107/S2056989019010557/su5505IVsup5.hklStructure factors: contains datablock(s) IV. DOI: 1942967, 1942966, 1942965, 1942964CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title heterocyclic 1,3,4-oxa\u00addiazole derivatives differ from each other in the groups attached to the carbon atoms: a meth\u00adoxy\u00adphenyl ring and a benzo\u00adnitrile group in (I) and a chloro\u00adphenyl ring and an acetamide group in (II). 18H15N3O4 and C17H14ClN3O3, are heterocyclic 1,3,4-oxa\u00addiazole derivatives which differ from each other in the groups attached to the carbon atoms: a meth\u00adoxy\u00adphenyl ring and a benzo\u00adnitrile group in (I) and a chloro\u00adphenyl ring and an acetamide group in (II). Short intra\u00admolecular C\u2014H\u22efO hydrogen bonds occur in both mol\u00adecules. The crystal structure of (I) features C\u2014H\u22efN hydrogen bonds, while in the crystal structure of (II), N\u2014H\u22efN, C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds are observed.The title compounds, C Compounds containing a heterocyclic ring system are of great importance both medicinally and industrially S(5) ring motifs and (II)s Figs. 1 and 2 \u25b8.B\u22efO3iii, Table\u00a01A\u22efN1i, Table\u00a01C(5) chains propagating along [010] chains propagating in an anti-parallel manner along [110]. These C\u2014H\u22efN hydrogen bonds along with the C\u2014H\u22efO dimers form a closed cavity shape arrangement consisting of 26 atoms in the unit cell if Fig.\u00a04. Further0] Fig.\u00a05. There ill Fig.\u00a06. In addiC(10) chains, C\u2014H\u22efN hydrogen bonds forming C(8) chains and C\u2014H\u22efO inter\u00adactions forming C(15) chains ns Fig.\u00a07. All thed Table\u00a02. No \u03c0\u2013\u03c0 2CO3 (3\u2005mmol) in DMF (4\u2005mL), 2-(chloro\u00admeth\u00adyl)-5-(4-meth\u00adoxy\u00adphen\u00adyl)-1,3,4-oxa\u00addiazole and KI (0.5\u2005mmol). The reaction mixture was stirred at room temperature for about 2\u2005h until the starting material had been consumed (TLC monitoring), and then washed with cold water. The solid product was collected by filtration and dried under vacuum. The pure compound was further recrystallized from ethyl acetate/petroleum ether solution (v:v = 1:1).Compound (I)N-(4-hy\u00addroxy\u00adphen\u00adyl)acetamide (1mmol), K2CO3 (3\u2005mmol) in ACN (5mL), 2-(chloro\u00admeth\u00adyl)-5-(4-chloro\u00adphen\u00adyl)-1,3,4-oxa\u00addiazole and KI (0.5\u2005mmol). The reaction mixture was stirred under reflux condition for about 16\u2005h, until completion of the reaction (TLC monitoring), then it was diluted with ethyl acetate (30\u2005mL) and washed with saturated NaHCO3 and cold water. The organic layer was separated, dried over anhydrous Na2SO4 and concentrated under vacuum. The pure compound was further recrystallized from an ethyl acetate/petroleum ether solution (v:v = 1:1), giving colourless block-like crystals suitable for X-ray diffraction analysis.Compound (II)Uiso(H) = 1.5Ueq(C-methyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018016754/zq2243sup1.cifCrystal structure: contains datablock(s) I, II, global. DOI: 10.1107/S2056989018016754/zq2243Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989018016754/zq2243IIsup4.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989018016754/zq2243Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018016754/zq2243IIsup5.cmlSupporting information file. DOI: 1881075, 1881074CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "K\u03b1 and Mo K\u03b1 radiation as part of a continuous crystallization study. In the crystal, chains along the a axis are formed via N\u2014H\u22efO hydrogen bonds between acetamide groups, as well as C\u2014H\u22efO inter\u00adactions. These chains arrange themselves into parallel running stacks which display weak C\u2014Cl\u22efO=Chalogen bonding as well as weak C\u2014H\u22ef\u03c0 inter\u00adactions.Two independent samples of the title compound were studied using Cu N-(4-methylphenyl)prop\u00adan\u00adamide, C10H12ClNO, 1, were studied using Cu K\u03b1, 1a, and Mo K\u03b1, 1b, radiation as part of a continuous crystallization study. The mol\u00adecule crystallizes with disorder in the Cl/terminal methyl positions [occupancies for the major disorder component of 0.783\u2005(2) in 1a and and 0.768\u2005(2) in 1b] and exhibits N\u2014C bond lengths of 1.3448\u2005(19), 1.344\u2005(2)\u2005\u00c5, C=O bond lengths of 1.2233\u2005(18) and 1.2245\u2005(19)\u2005\u00c5 and an acetamide moiety C\u2014N\u2014C\u2014C torsion angle of 179.00\u2005(13), 178.97\u2005(14) \u00b0 for 1a and 1b, respectively. In the crystal, chains along the a axis are formed via N\u2014H\u22efO hydrogen bonds between acetamide groups, as well as C\u2014H\u22efO inter\u00adactions. These chains arrange themselves into parallel running stacks which display weak C\u2014Cl\u22efO=C halogen bonding as well as weak C\u2014H\u22ef\u03c0 inter\u00adactions.Two independent samples of the title compound, alternatively 2-chloro- The disorder occupancy is different in 1a, 1b and in IQOHOL, but to no great extent with the occupancy of the major component being 0.783\u2005(2) for 1a, 0.768\u2005(2) for 1b and for 0.899 IQOHOL.An overlay of the mol\u00adecular structures of i, see Tables 2et al., 20011.In the extended structure there is, as expected, a strong amide hydrogen bond, between the N\u2014H group and the ketone oxygen and 2.8632\u2005(6)\u2005\u00c5 respectively)] The head-to-tail packing and the chelate hydrogen bonding allows an approximately linear arrangement of 1, forming ribbons propagating along the [100] direction, see Fig.\u00a031a and 1.71940\u2005(13)\u00b0 in 1b).There is also a weaker inter\u00adaction between the methine group and the ketone and 1b, 3.1734\u2005(18)\u2005\u00c5 . A very weak example of a C\u2014H\u22ef\u03c0iii inter\u00adaction is also present in 1, with the methyl group C12 directed towards the centroid of ring C1\u2013C6 yielded several similar substituted phenyl\u00adacetamides: CLACTN in toluene (50\u2005mL) and 40\u2005mL of aqueous NaOH at 273\u2005K. After the addition was complete, the biphasic suspension was warmed to room temperature and stirred vigorously for 1\u2005h. The organic phase was separated, and the aqueous layer extracted with ethyl acetate (3 \u00d7 15\u2005mL). The organic layers were then combined, dried with Na2SO4, filtered and the solvent removed under vacuum. The resulting off-white solid was collected and washed with thoroughly with cold cyclo\u00adhexane . Single crystals for X-ray analysis were grown by slow evaporation of a toluene solution at room temperature. Spectroscopic data for the obtained product matched that reported in the literature in toluene (30\u2005mL) was added dropwise (with extreme caution) to a vigorously stirred bi-phasic suspension of 1H NMR : \u03b4 8.21 , 7.42 , 7.15 , 4.54 2.13 , 1.83 . 13C NMR : \u03b4 166.9 134.4, 134.0, 129.1, 119.7, 55.9, 22.4, 20.5. MS (EI) m/z 197 [M]+, [12C10H1235Cl14N16O 197]. HRMS (EI) m/z Found: [M]+ 197.0604, [C10H12ClNO]+ requires 197.0607.1a and 1b, Cl1/Cl1a and C12/C12a were modelled as disordered over two positions using restraints and constraints . The occupancy was allowed to refine with a population parameter of 1a = 0.783\u2005(2), and 1b = 0.768\u2005(2). The amide N\u2014H H atom was located in a difference-Fourier map and freely refined. H atoms bonded to carbon were placed with idealized geometry and refined using a riding model with C\u2014H = 0.95\u2005\u00c5 aromatic, C\u2014H = 0.90\u2005\u00c5 methine, with Uiso(H) = 1.2Ueq(C) and C\u2014H = 0.98\u2005\u00c5 methyl with Uiso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989018013889/ds2252sup1.cifCrystal structure: contains datablock(s) . DOI: 10.1107/S2056989018013889/ds22521asup2.hklStructure factors: contains datablock(s) 1a. DOI: 10.1107/S2056989018013889/ds22521bsup3.hklStructure factors: contains datablock(s) 1b. DOI: Click here for additional data file.10.1107/S2056989018013889/ds22521asup4.cmlSupporting information file. DOI: 1870782, 1870781CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure of the title sulfonamide, inter\u00admolecular N\u2014H\u22efO hydrogen bonds are present between sulfonamide groups, as well as offset \u03c0\u2013\u03c0 inter\u00adactions. 10H13NO2S, was synthesized by a nucleophilic substitution reaction between allyl amine and p-toluene\u00adsulfonyl chloride. The sulfonate S\u2014O bond lengths are 1.4282\u2005(17) and 1.4353\u2005(17)\u2005\u00c5, and the C\u2014N\u2014S\u2014C torsion angle involving the sulfonamide moiety is \u221261.0\u2005(2)\u00b0. In the crystal, centrosymmetric dimers of the title compound are present via inter\u00admolecular N\u2014H\u22efO hydrogen bonds between sulfonamide groups. These dimers are linked into ribbons along the c-axis direction through offset \u03c0\u2013\u03c0 inter\u00adactions.The title compound, C Centrosymmetric dimers of compound (I)i distance of 2.900\u2005(3)\u2005\u00c5 suggests inter\u00adactions of medium strength with a nearly linear N\u2014H\u22efO hydrogen bond of 174\u2005(3)\u00b0 \u2005\u00c5, with a slippage of 1.320\u2005\u00c5 and a plane-to-plane distance between phenyl rings of 3.600\u2005\u00c5 .Mol\u00adecules of the title compound are linked to one another \u00b0 Table\u00a01. These ds Figs. 3, 4 \u25b8. Thet al., 2016p-tolyl\u00adsulfonamides where there is a \u2013CH2\u2014C=C group bonded to the sulfonamide-N atom. The alkene group in these structures is a part of, for example, furan rings . The mixture was stirred at room temperature for 24\u2005h. Reaction completion was verified by using TLC analysis. The mixture was acidified to pH 2\u20133 using concentrated HCl. After dilution with 20\u2005ml of CH2Cl2, the organic phase was washed with H2O (3 \u00d7 20\u2005ml) and the aqueous layer was back-extracted with CH2Cl2 (20\u2005ml). The combined organic extracts were dried over anhydrous Na2SO4. After solvent evaporation, the residue was obtained as a yellow solid which was recrystallized in cold ethanol to afford pale-yellow crystals .Allyl\u00adamine was added in 20\u2005ml of degassed di\u00adchloro\u00admethane. This was followed by the addition of pyridine . The resulting solution was stirred under an atmosphere of Nsp3\u2014H = 0.95\u20131.00\u2005\u00c5 with Uiso(H) = 1.2Ueq(C) for methine and methyl\u00adene groups, and Uiso(H) = 1.5Ueq(C) for methyl groups. The hydrogen atom bonded to the nitro\u00adgen atom (H1) was located using electron-density difference maps, and the N\u2014H bond length was restrained to 0.84\u00b10.01\u2005\u00c5 using the DFIX command as executed in SHELXL I. DOI: 10.1107/S2056989018010290/wm5455Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018010290/wm5455Isup3.cmlSupporting information file. DOI: 1856234CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Z\u2032 = 1 is reported.A new monoclinic polymorph of 2,2\u2032-methyl\u00adenebis with C2/c) of 2,2\u2032-methyl\u00adenebis, C17H10N2O4, is reported and compared to the previously reported triclinic polymorph (space group PZ\u2032 = 1). The mol\u00adecular conformations of the two polymorphs are very similar, as shown by the r.m.s. deviation of 0.368\u2005\u00c5 . The inter\u00admolecular inter\u00adactions of both polymorphs are described along with the Hirshfeld surface analysis, and the lattice energies are calculated.In this study, a new monoclinic polymorph (space group N-heterocycles have been proven to exhibit significant biological and pharmaceutical activities, and have also been used as dyes and heat-resistant polymers in industry \u2005\u00c5, b = 9.5810\u2005(8)\u2005\u00c5, c = 10.2780\u2005(6)\u2005\u00c5, \u03b1 = 104.325\u2005(3)\u00b0, \u03b2 = 99.768\u2005(4)\u00b0, \u03b3 = 96.030\u2005(3)\u00b0, Z = 2, Z\u2032 = 1 and V = 712.23\u2005(11)\u2005\u00c53; Cambridge Structural Database of 2,2\u2032-methyl\u00adenebis with Z\u2032 = 1 and compare its properties with those of 1\u03b1. According to the Online Dictionary of Crystallography, polymorphism is the phenomenon in which the same chemical compound exhibits different crystal structures derivatives with five-membered \u03b1 and 1\u03b2 and 0.064\u2005(3)\u2005\u00c5 for 1\u03b2. There are two degrees of freedom to characterize the mol\u00adecular conformations of 1\u03b1 and 1\u03b2: these are the torsion angles C1\u2014N1\u2014C9\u2014N2 and N1\u2014C9\u2014N2\u2014C10 . Generally, the mol\u00adecule of 1\u03b2 deviates only slightly from that of 1\u03b1, as indicated by a r.m.s. deviation of 0.368\u2005\u00c5 \u00b0, which is smaller than that of 88.96\u2005(4)\u00b0 observed in polymorph 1\u03b1. The calculated density and Kitaigorodskii packing index are slightly higher than those observed for 1\u03b1 (1.428 Mg\u2005m\u22123 and 69.0%).The asymmetric units of polymorphs 11\u03b2 Fig.\u00a01 each cons) Fig.\u00a02. The mea\u03b1 features weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 inter\u00adactions between neighboring phthalimide units. In the crystal structure of 1\u03b2 and between N1/C1/C2/C7/C8 and C11\u2013C16 rings and 1\u03b2 ], respectively. The percentages of contribution of H\u22efC/C\u22efH, O\u22efC/C\u22efO and C\u22efC contacts to the Hirshfeld surface are 20.6, 3.3 and 8.9%, respectively, for 1\u03b1, and 20.8, 7.9 and 6.7%, respectively, for 1\u03b2 ]. The C\u22efC contacts appear as a unique \u2018triangle\u2019 focused at de \u2243 di \u2243 1.75\u2005\u00c5 [Fig.\u00a04f)]. The inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions are illustrated as unique patterns of red and blue \u2018triangles\u2019 on the shape-index surface is slightly larger than for 1\u03b2 (128.5\u2005kJ\u2005mol\u22121), indicating that 1\u03b1 is slightly more stable than 1\u03b2 under ambient conditions.The C\u2014H bond lengths in 1\u03b2 were obtained from an unsuccessful synthesis of 2-{[(3-iodo\u00adpyridin-4-yl)amino]\u00admeth\u00adyl}isoindoline-1,3-dione by reacting N-(bromo\u00admeth\u00adyl)phthalimide (1\u2005mmol) and 4-amino-3-iodo\u00adpyridine (1\u2005mmol) in N,N-di\u00admethyl\u00adformamide (8\u2005ml) with the presence of a catalytic amount of anhydrous potassium carbonate. The reaction solution was stirred for about 2\u2005h at room temperature. Once the reaction was complete, the resultant mixture was poured into a beaker of ice-cooled water to obtain a precipitate = 1.2Ueq(C).Crystal data, data collection and structure refinement details of 110.1107/S2056989018017425/jj2205sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018017425/jj2205Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018017425/jj2205Isup3.cmlSupporting information file. DOI: 1884044CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound is a salt formed by a hydrazinium (1+) cation and a hexa\u00adfluorido\u00adsilicate anion inter\u00adconnected by N\u2014H\u22efN and N\u2014H\u22efF hydrogen bonds. 2H5)2SiF6, the silicon atom at the centre of the slightly distorted SiF6 octa\u00adhedron [range of Si\u2014F distances = 1.6777\u2005(4)\u20131.7101\u2005(4)\u2005\u00c5] lies on a crystallographic inversion centre. In the crystal, the ions are connected by N\u2014H\u22efN and N\u2014H\u22efF hydrogen bonds; the former link the cations into [010] chains and the latter (some of which are bifurcated or trifurcated) link the ions into a three-dimensional network. The two-dimensional fingerprint plots show that F\u22efH/H\u22efF inter\u00adactions dominate the Hirshfeld surface (75.5%) followed by H\u22efH (13.6%) and N\u22efH/H\u22efN (8.4%) whereas F\u22efF (1.9%) and F\u22efN/N\u22efF (0.6%) have negligible percentages. The title compound is isostructural with its germanium-containing analogue.In the title inorganic mol\u00adecular salt, (N However, this compound was characterized by chemical analysis, vibrational spectroscopy and X-ray powder photography by Gantar & Rahten to 1.7101\u2005(4)\u2005\u00c5. The minimum and maximum cis F\u2014S\u2014F angles are 89.26\u2005(2) and 90.74\u2005(2)\u00b0, respectively. The N\u2014N separation in the cation is 1.4416\u2005(8)\u2005\u00c5.Compound (I)6]2\u2212 anion inter\u00adacts with the (N2H5)+ cations through electrostatic attraction and accepts no fewer than ten simple, bifurcated or trifurcated N\u2014H\u22efF hydrogen bonds ds Fig.\u00a03. This reon Fig.\u00a04.et al., 2004CrystalExplorer package dnorm . In order to provide qu\u00adanti\u00adtative information on the contribution of the inter\u00admolecular inter\u00adactions to the crystal packing, the three-dimensional dnorm surface is resolved into two-dimensional fingerprint plots, generated based on de and di distance scales and illustrated in Fig.\u00a05a)\u2013(f) The F\u22efH/H\u22efF inter\u00adactions appear as distinct spikes in the fingerprint plot, and occupy the majority of the total Hirshfeld surface (75.5%) as illustrated in Fig.\u00a05a); the characteristic \u2018wingtip\u2019 features indicate the N\u2014H\u22efF hydrogen bonds. The H\u22efF inter\u00adaction are represented by a spike at the bottom left (donor), whereas the F\u22efH inter\u00adactions are represented by a spike at the bottom right (acceptor) of the fingerprint plot. The H\u22efH contacts appear in the middle of the scattered points; these contacts comprise 13.6% of the total Hirshfeld surface [Fig.\u00a05c)]. The N\u22efH contacts cover 8.4% of the total surface, as the third important contributor in the crystal packing, Fig.\u00a05d) while the F\u22efF and F\u22efN/N\u22efF contacts make negligible contributions of 1.9% [Fig.\u00a05e)] and 0.6% [Fig.\u00a05f)], respectively.The acceptor atoms in the inter\u00adactions are shown with negative electrostatic potentials (red regions), and donor atoms are shown with positive electrostatic potentials (blue regions). The N\u2014H\u22efF inter\u00adactions in the structure are apparent from the relatively bright red-spots on the Hirshfeld surface of (I)2H6SiF6, at room temperature, crystallizes in a pseudo-tetra\u00adgonal ortho\u00adrhom\u00adbic space group , with a = 7.605\u2005(1)\u2005\u00c5, b = 7.586\u2005(2)\u2005\u00c5 and c = 8.543\u2005(1)\u2005\u00c5 plane.Hydrazinium (2+) hexa\u00adfluorido\u00adsilicate, Net al., 19852H5)2GeF6 crystallizes in the monoclinic system, space group P21/n (Z = 2), with cell parameters a = 6.015\u2005(2)\u2005\u00c5, b = 5.249\u2005(1)\u2005\u00c5, c = 11.181\u2005(2)\u00c5 and \u03b2 = 100.15\u2005(2)\u00b0 and is clearly isostructural with (I)Hydrazinium (1+) hexa\u00adhalogenometallates were studied by Gantar and co-workers (Gantar 2H5)2TiF6 2TiF6 exhibit racemic twinning but are not isostructural with (I)2H5)2TiF6 consists of N2H5+ cations and two types of slightly distorted octa\u00adhedral (TiF6)2\u2212 anions. The N2H5+ cations and (TiF6)2\u2212 anions are linked via N\u2014H\u22efF and N\u2014H\u22efN hydrogen bonds, building a three-dimensional network. Two other isostructural hydrazinium (l+) hexa\u00adfluorido complexes, (N2H5)2ZrF6 and (N2H5)2HfF6, were prepared and characterized by chemical analysis, vibrational spectroscopy and X-ray powder diffraction .Fluoride complexes of titanium (IV) with ammonium cation derivatives include two hydrazinium hexa\u00adfluorido\u00adtitanates (IV), (N2H5)2SiF6 crystals in the form of colourless blocks were obtained by slow evaporation, at room temperature, of an aqueous solution containing stoichiometric amounts of hydrazine NH2NH2 and H2SiF6. The infrared spectrum was recorded in the range 450\u20134000\u2005cm\u22121 with a Vertex 70 FTIR spectrometer.Hydrazinium (1+) hexa\u00adfluorido\u00adsilicate (NUiso(H) = 1.2Ueq(N). The highest peak and the deepest hole in the final Fourier map are at 0.67\u2005\u00c5 from F3 and 0.0\u2005\u00c5 from Si1.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019012672/hb7850sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019012672/hb7850Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019012672/hb7850sup3.pdfSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019012672/hb7850sup3.rtfSupporting information file. DOI: 1953037CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The syntheses and crystal structures of three cyclo\u00adtriphosphazenes, all with fluorinated ar\u00adyloxy side groups that generate different steric characteristics are reported. viz. hexa\u00adkis\u00ad(penta\u00adfluoro\u00adphen\u00adoxy)cyclo\u00adtriphosphazene, N3P3(OC6F5)6, 1, hexa\u00adkis\u00ad[4-(tri\u00adfluoro\u00admethyl)\u00adphen\u00adoxy]cyclo\u00adtriphosphazene, N3P3[OC6H4(CF3)]6, 2 and hexa\u00adkis\u00adcyclo\u00adtriphosphazene, N3P3[OC6H3(CF3)2]63, are reported. Specifically, each phospho\u00adrus atom bears either two penta\u00adfluoro\u00adphen\u00adoxy, 4-tri\u00adfluoro\u00admethyl\u00adphen\u00adoxy, or 3,5-tri\u00adfluoro\u00admethyl\u00adphen\u00adoxy groups. The central six-membered phosphazene rings display envelope pucker conformations in each case, albeit to varying degrees. The maximum displacement of the \u2018flap atom\u2019 from the plane through the other ring atoms [0.308\u2005(5)\u2005\u00c5] is seen in 1, in a mol\u00adecule that is devoid of hydrogen atoms and which exhibits a \u2018wind-swept\u2019 look with all the aromatic rings displaced in the same direction. In 3 an intra\u00admolecular C\u2014H(aromatic)\u22efF inter\u00adaction is observed. All the \u2013CF3 groups in 2 and 3 exhibit positional disorder over two rotated orientations in close to statistical ratios. The extended structures of 2 and 3 are consolidated by C\u2014H\u22efF inter\u00adactions of two kinds: (a) linear chains, and (b) cyclic between mol\u00adecules related by inversion centers. In both 1 and 3, one of the six substituted phenyl rings has a parallel-displaced aromatic \u03c0\u2013\u03c0 stacking inter\u00adaction with its respective symmetry mate with slippage values of 2.2\u2005\u00c5 in 1 and 1.0\u2005\u00c5 in 3. None of the structures reported here have solvent voids that could lead to clathrate formation.The syntheses and crystal structures of three cyclo\u00adtriphosphazenes, all with fluorinated ar\u00adyloxy side groups that generate different steric characteristics, Cyclic organophosphazenes have a long history as representatives of inorganic heterocyclic rings, and are also the focus of arguments about reactivity and pseudoaromaticity in inorganic systems 6 (1), hexa\u00adkis\u00ad[4-(tri\u00adfluoro\u00admethyl)\u00adphenoxy]cyclo\u00adtriphosphazene N3P3[OC6H4(CF3)]6 (2) and hexa\u00adkis\u00ad6 (3) were synthesized by the reactions of hexa\u00adchloro\u00adcyclo\u00adtriphosphazene with the appropriate sodium fluoro-aryl\u00adoxides in THF or dioxane solvent. Forcing reaction conditions (boiling dioxane) were required for complete chlorine replacement in the case of the penta\u00adfluoro\u00adphen\u00adoxy derivative (1) presumably due to steric hindrance. The three compounds were characterized by NMR spectroscopy in addition to x-ray crystallography.As part of our ongoing work in this area, the three cyclo\u00adtriphosphazenes hexa\u00adkis\u00ad(penta\u00adfluoro\u00adphen\u00adoxy)cyclo\u00adtri\u00adphos\u00adphazene Npara-fluoro\u00adphen\u00adoxy groups, was reported by other investigators is 0.308\u2005(5)\u2005\u00c5. A similar calculation in 2 shows atom P2 displaced by 0.232\u2005(4)\u2005\u00c5, and in 3 atom N3 is displaced the least, only by 0.205\u2005(4)\u2005\u00c5. In earlier structure reports: .The mol\u00adecular structure of ce Fig.\u00a04 with all1, hydrogen bonding is not feasible in that structure \u22efF type hydrogen bonds \u22efF hydrogen bonds \u2005\u00c5, slippage 1.013\u2005\u00c5].The packing diagram of s Table\u00a02, forminget al. 3, CSD refcode KAGKUY: Bullen, 19712)3, VARYES02: Singh et al., 20002)3\u00b7THF, GUHPII: Dietrich et al., 2000Earlier, we reported the crystal structures of a number of cyclo\u00adtriphosphazenes with spiro\u00adcyclic ar\u00adyloxy side groups cyclo\u00adtriphos\u00adpha\u00adzene (1):4, and concentrated to \u223c10\u2005ml by rotary evaporation. A small amount of hexa\u00adnes was added to the concentrated solution and it was chilled to 273\u2005K via an ice bath to yield colorless blocks of 1, which were filtered and rinsed with cold hexa\u00adnes then dried under vacuum.Sodium penta\u00adfluoro\u00adphenoxide was prepared by the treatment of penta\u00adfluoro\u00adphenol with a suspension of NaH 60% dispersion in mineral oil in 50\u2005ml of dioxane. The penta\u00adfluoro\u00adphenoxide was added to a stirred solution of hexa\u00adchloro\u00adcyclo\u00adtriphosphazene and the mixture was heated at reflux for 3\u2005d. Dioxane was removed from the mixture by rotary evaporation and the residue was dissolved in 100\u2005ml di\u00adchloro\u00admethane. The solution was extracted with 3 \u00d7 100\u2005ml of deionized water, dried over MgSOSynthesis of hexa\u00adkis\u00ad(4-tri\u00adfluoro\u00admethyl\u00adphen\u00adoxy)cyclo\u00adtri\u00adphos\u00adphazene (2):1 to yield colorless cubes of 2.The aryl\u00adoxide was prepared by treatment of 4-tri\u00adfluoro\u00admethyl\u00adphenol with a suspension of NaH [60% dispersion in mineral oil ] in 50\u2005ml of THF. To the stirred solution of 4-tri\u00adfluoro\u00admethyl\u00adphenoxide was added a solution of hexa\u00adchloro\u00adcyclo\u00adtriphosphazene in 15\u2005ml of THF and the mixture was stirred at room temperature overnight. The purification steps of this compound were identical to those of compound Synthesis of hexa\u00adkis\u00adcyclo\u00adtriphosphazene (3):4 and the di\u00adchloro\u00admethane was removed by rotary evaporation to yield a colorless oil, which crystallized as colorless needles of 3 after standing for several hours. The crystals were rinsed with cold methanol and then dried under vacuum.A stirred suspension of NaH [60% dispersion in mineral oil ] in 25\u2005ml of THF was treated with liquid 3,5-bis-tri\u00adfluoro\u00admethyl\u00adphenol by dropwise addition. The resulting aryl\u00adoxide solution was then added to a stirred solution of hexa\u00adchloro\u00adcyclo\u00adtriphosphazene in 25\u2005ml of THF and the reaction mixture was stirred at room temperature overnight. The mixture was concentrated by rotary evaporation and the residue was dissolved in 40\u2005ml di\u00adchloro\u00admethane. The di\u00adchloro\u00admethane solution was washed with 40\u2005ml of deionized water, followed by 20\u2005ml of 5% HCl, and finally rinsed with 40\u2005ml of deionized water. The organic layer was dried over MgSO1\u20133 are as follows: 1: 31P 10.6 1 ppm (in chloro\u00adform-d); 19F \u2212153.52 , \u2212157.53 , \u2212161.79 ; 2: 31P 8.64 ppm (in chloro\u00adform-d); 1H 747, 706 (d 2H); 19F \u221262.79 (s); 3: 31P 7.71 ppm (in chloro\u00adform-d); 1H 7.74 (s 1H), 7.52 ; 19F \u221263.85 (s).The NMR data for 2 and 3 were placed geometrically (C\u2014H = 0.93\u2005\u00c5) and refined as riding on their parent atoms with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details for all three structures are summarized in Table\u00a0310.1107/S2056989019012933/hb7853sup1.cifCrystal structure: contains datablock(s) 1, 2, 3. DOI: 10.1107/S2056989019012933/hb78531sup2.hklStructure factors: contains datablock(s) 1. DOI: Click here for additional data file.10.1107/S2056989019012933/hb78531sup5.molSupporting information file. DOI: 10.1107/S2056989019012933/hb78532sup4.hklStructure factors: contains datablock(s) 2. DOI: Click here for additional data file.10.1107/S2056989019012933/hb78532sup6.molSupporting information file. DOI: 10.1107/S2056989019012933/hb78533sup3.hklStructure factors: contains datablock(s) 3. DOI: Click here for additional data file.10.1107/S2056989019012933/hb78533sup7.molSupporting information file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Enterobacteriaceae family were particularly most sensitive to new pipemidic acid derivatives. Synthesized compounds exhibited very strong activity towards Proteus mirabilis ATCC 12453, Salmonella typhimurium ATCC 14028 and Escherichia coli ATCC 25922. The minimum inhibitory concentrations of new pipemidic acid derivatives which inhibited the growth of these bacteria were 0.98\u20137.81\u00a0\u00b5g/ml, 0.98\u20137.81\u00a0\u00b5g/ml and 0.98\u20133.91\u00a0\u00b5g/ml, respectively. The antibacterial activity of newly synthesized pipemidic acid derivatives in many cases was far better than the activity of substances used as positive controls .This article describes the synthesis and antimicrobial activity evaluation of new pipemidic acid derivatives. New compounds were obtained on the basis of Mannich reaction of 4,5-disubstituted 1,2,4-triazole-3-thiones with pipemidic acid. Antimicrobial tests revealed high antibacterial activity of obtained derivatives. Gram-negative rods belonging to The online version of this article (10.1007/s12272-018-1025-3) contains supplementary material, which is available to authorized users. Microorganisms play a key role in the functioning of the environment but they are also a real threat to human life and health. The widespread diseases and infections caused by different bacteria and fungi have encourage the need for scientific research and advances in medicinal chemistry . The DMSO-d6 was used as solvent and TMS as the internal standard. Chemical shifts in this article are reported in ppm (\u03b4). The coupling constants (J) are presented in Hertz. The purity of obtained compounds and the progress of the reaction were determined by thin-layer chromatography (TLC) with the use of pre-coated aluminum sheet 60 F254 plates (Merck Co. USA), and CHCl3/C2H5OH solvent system. The spots were identified by the exposure to the UV light at 254\u00a0nm. The elemental analysis of obtained compounds was carried out with the use of AMZ 851 CHX analyser . The results of elemental analysis were within\u2009\u00b1\u20090.4% of the calculated values.The reagents and solvents used in this research were obtained from Merck Co. and Sigma-Aldrich . Melting points were determined with the use of Fisher-Johns blocks melting point apparatus and presented without correction. The 1 or 4-tert-butylbenzhydrazide\u20142, respectively) was dissolved in 10\u00a0ml of ethanol (96%). Then 0.0022\u00a0mol of appropriate isothiocyanate was added and heated under reflux for 3\u00a0h. Subsequently obtained solution was put to the refrigerator for 24\u00a0h. After that formed precipitate was filtered off and re-crystallized from ethanol. The procedure of this synthesis was based on our previous article is presented in Supplementary Materials.Detailed physicochemical data of thiosemicarbazide derivatives (3\u201318) was dissolved in 5\u00a0ml of 2% sodium hydroxide solution and heated under reflux for 2\u00a0h. Subsequently, obtained solution was neutralized with diluted hydrochloric acid. Formed precipitate was filtered off and re-crystallized from ethanol.4,5-Disubstituted 1,2,4-triazole-3-thiones were synthesized with the use of the procedure from our earlier research is presented in Supplementary Materials.Detailed physicochemical data of 4,5-disubstituted 1,2,4-triazole-3-thione derivatives (19\u201334) was added to the conical flask and dissolved with stirring in 5\u00a0ml of ethanol (96%). After that 200\u00a0\u00b5l of formaldehyde and 0.0033\u00a0mol of pipemidic acid was added to the flask. The content of the flask was stirred by magnetic stirrer for 1\u00a0h. Subsequently, 15\u00a0ml of distilled water was added to the flask. The precipitate which formed was filtered off and re-crystallized from methanol.In order to obtained new pipemidic acid derivatives we applied the Mannich reaction and the procedure reported by our group earlier is presented in Supplementary Materials.Detailed physicochemical data of new pipemidic acid derivatives (EUCAST discussion document E. Dis 5.1 The examined compounds 35\u201350) were obtained with the use of three step reaction scheme (Scheme\u00a03\u201318) were synthesized on the basis of condensation reaction of appropriate carboxylic acid hydrazides with various isothiocyanates. Subsequently, thiosemicarbazide derivatives (3\u201318) underwent cyclization reaction with the use of 2% sodium hydroxide solution, which afforded the 4,5-disubstituted 1,2,4-triazole-3-thione derivatives (19\u201334). Finally, 1,2,4-triazole-3-thione derivatives (19\u201334) were subjected to Mannich reaction with pipemidic acid to obtain new pipemidic acid derivatives (35\u201350). Chemical structure of all obtained compounds (3\u201350) was confirmed on the basis of spectral identification and elemental analysis.New pipemidic acid derivatives (e Scheme\u00a0. Firstly3\u201318), 1,2,4-triazoles-3-thiones (19\u201334) and pipemidic acid derivatives (35\u201350), were in vitro screened against references strains of bacteria and fungi. The results of antimicrobial assays are presented in the Tables\u00a0All synthesized compounds, including thiosemicarbazides 3\u20138, 1,2,4-3\u201318) defined as MIC values against Gram-positive bacterial strains ranged from 3.91\u00a0\u00b5g/ml to 1000\u00a0\u00b5g/ml , against Gram-negative bacterial strains ranged from 500\u00a0\u00b5g/ml to 1000\u00a0\u00b5g/ml (MBC\u2009=\u2009>\u20091000\u00a0\u00b5g/ml). The MIC values against reference fungi were MIC\u2009=\u2009250\u20131000\u00a0\u00b5g/ml were within the range of 500\u20131000\u00a0\u00b5g/ml against Gram-positive bacterial strains (MBC\u2009=\u20091000 to >\u20091000\u00a0\u00b5g/ml), 1000\u00a0\u00b5g/ml against Gram-negative bacterial strains (MBC\u2009=\u2009>\u20091000\u00a0\u00b5g/ml) and 250\u20131000\u00a0\u00b5g/ml for fungal strains (MBC\u2009=\u20091000 to >\u20091000\u00a0\u00b5g/ml) ranged from 3.91 to 1000\u00a0\u00b5g/ml against Gram-positive bacteria (MBC\u2009=\u20093.91 to >\u20091000\u00a0\u00b5g/ml), 0.98\u2013125\u00a0\u00b5g/ml against Gram-negative bacteria (MBC\u2009=\u20090.98\u2013250\u00a0\u00b5g/ml) and 1000\u00a0\u00b5g/ml towards fungi belonging to Candida spp. (MBC\u2009=\u2009>\u20091000\u00a0\u00b5g/ml) , 4,5-disubstituted 1,2,4-triazoles-3-thiones (19\u201334) and pipemidic acid derivatives (35\u201350) in this research was confirmed on the basis of spectral (1H NMR and 13C NMR) and elemental analysis.The pathway for the synthesis of new pipemidic acid derivatives Scheme\u00a0 was desi3\u201318) showed three typical singlet signals at \u03b4 7.99\u201310.52\u00a0ppm, on the 1H NMR spectra, which correspond to three NH groups. Whereas on the 13C NMR spectra of this group of compounds (3\u201318), signals for carbonyl group (C=O) and thiocarbonyl group (C=S) were found at \u03b4 155.1\u2013159.5\u00a0ppm and around \u03b4 166.0\u00a0ppm, respectively.Thiosemicarbazide derivatives (19\u201334), on the 1H NMR spectra, singlet signal for NH group was noticed in the range of \u03b4 13.75\u201314.15\u00a0ppm. On the 13C NMR spectra the signal for thiocarbonyl group (C=S) of 1,2,4-triazole-3-thiones (19\u201334) was found at \u03b4 166.5\u2013169.3\u00a0ppm.In the case of 4,5-disubstituted 1,2,4-triazole-3-thione derivatives (1H NMR spectra of new pipemidic acid derivatives (35\u201350) showed characteristic singlet signal for CH2 group in the range of \u03b4 5.18\u20135.36\u00a0ppm, what confirmed common aminomethylation reaction of 1,2,4-triazole-3-thiones. Signals for other aliphatic and aromatic fragments of synthesized compounds on 1H NMR and 13C NMR were shown at expected shift range.The 35\u201350) and six compounds among 4,5-disubstituted 1,2,4-triazole derivatives (19\u201334) are new in the literature and their synthesis, physicochemical data and biological activity have not been reported so far.It is worth to mention that all sixteen synthesized pipemidic acid derivatives (3\u201318), especially compounds: 4\u201310, 12, 13, 15, 18, had no activity against all reference microorganisms values, was 32 times better against M. luteus ATCC 10240 (MIC\u2009=\u20093.91\u00a0\u00b5g/ml) and two times better against B. cereus ATCC 10876 (MIC\u2009=\u20097.81\u00a0\u00b5g/ml) in comparison with the activity of pipemidic acid , used as positive control and moderate or mild effect against other reference staphylococci and some Gram-negative rods (MIC\u2009=\u2009250\u20131000\u00a0\u00b5g/ml and MBC\u2009>\u20091000\u00a0\u00b5g/ml). Moreover, the compounds 3, 11, 14 and 16 exhibited mild activity (MIC\u2009=\u20091000\u00a0\u00b5g/ml and MBC\u2009>\u20091000\u00a0\u00b5g/ml) towards some Gram-positive bacteria. The substances 3 and 11 showed additional effect against Bordetella bronchiseptica ATCC 4617 (MIC\u2009=\u2009500\u20131000\u00a0\u00b5g/ml and MBC\u2009>\u20091000\u00a0\u00b5g/ml) , only a few, namely 19, 20, 21 and 34 exhibited moderate or mild antibacterial activity against some of reference microorganisms . The remaining substances 22\u201333 were inactive towards bacteria and fungi from ATCC were highly active against all reference bacteria (Table\u00a0Enterobacteriaceae family were particularly most sensitive to these compounds. All substances (35\u201350) showed bactericidal effect against them. These compounds exhibited very strong activity towards Proteus mirabilis ATCC 12453, Salmonella typhimurium ATCC 14028 and Escherichia coli ATCC 25922. The minimum concentrations of 35\u201350 compounds, which inhibited the growth of these bacteria were 0.98\u20137.81\u00a0\u00b5g/ml, 0.98\u20137.81\u00a0\u00b5g/ml and 0.98\u20133.91\u00a0\u00b5g/ml, respectively. Klebsiella pneumoniae ATCC 13883 was slightly less susceptible to these substances. The compounds 36, 37, 38, 44, 46 and 50 indicated very strong activity (MIC\u2009=\u2009MBC\u2009=\u20093.91\u20137.81\u00a0\u00b5g/ml), compounds 35, 39 and 45\u2014strong , while remaining compounds 40, 41, 42, 43, 47, 48, 49\u2014good activity (MIC\u2009=\u2009MBC\u2009=\u200931.25\u201362.5\u00a0\u00b5g/ml) towards reference K. pneumoniae ATCC 13883. In addition, all pipemidic acid derivatives (35\u201350) exhibited a similar good effect against Bordetella bronchiseptica ATCC 4617 and Pseudomonas aeruginosa ATCC 9027 towards B. bronchiseptica ATCC 4617 were two times better in case of compounds 36, 37, 45, 46 and 50 (MBC\u2009=\u200931.25\u00a0\u00b5g/ml). Against K. pneumoniae ATCC 13883, the compound 36 showed four times better activity (MIC\u2009=\u20093.91\u00a0\u00b5g/ml), compounds 37, 38, 44, 46, and 50 showed two times better activity (MIC\u2009=\u20097.81\u00a0\u00b5g/ml) than pipemidic acid (MIC\u2009=\u200915.62\u00a0\u00b5g/ml). The compounds 35 and 45 showed two times lower MBC values (MBC\u2009=\u200915.62\u00a0\u00b5g/ml) than pipemidic acid (MBC\u2009=\u200931.25\u00a0\u00b5g/ml) against this bacterium. In the case of the activity against P. mirabilis ATCC 12453 the MIC values for the compound 36 were two times lower (MIC\u2009=\u20090.98\u00a0\u00b5g/ml) and the MBC values for the compounds 35, 38, 46 were two times lower (MBC\u2009=\u20091.95\u00a0\u00b5g/ml) than such values for pipemidic acid . The activity of synthesized derivatives was also better than pipemidic acid towards Salmonella typhimurium ATCC 14028. The compounds 37 and 38 showed two times better activity on the basis of MIC values (MIC\u2009=\u20090.98\u00a0\u00b5g/ml) than pipemidic acid (MIC\u2009=\u20091.95\u00a0\u00b5g/ml) against this bacterium. The MBC values for compounds 35, 36, 45 and 46 (MBC\u2009=\u20091.95\u00a0\u00b5g/ml) were two times lower than for pipemidic acid used as positive control (MBC\u2009=\u20093.91\u00a0\u00b5g/ml). In addition to this, the MBC values for compounds 36, 37, 38, and 45 against E. coli ATCC 25922 (MBC\u2009=\u20090.98\u00a0\u00b5g/ml) were two times lower than for pipemidic acid (MBC\u2009=\u20091.95\u00a0\u00b5g/ml) values of pipemidic acid derivatives 5\u201350 towa35-50 indicated also high activity towards Gram-positive bacteria but slightly weaker compared to Gram-negative microorganisms. Most of the substances showed very strong bactericidal or bacteriostatic effect against Bacillus subtilis ATCC 6633 with MIC\u2009=\u20093.91\u20137.81\u00a0\u00b5g/ml, MBC\u2009=\u20093.91\u2013250\u00a0\u00b5g/ml and MBC/MIC\u2009=\u20091\u201332. The other compounds 43 and 49 indicated good (MIC\u2009=\u2009MBC\u2009=\u200931.25\u00a0\u00b5g/ml) or strong (MIC\u2009=\u2009MBC\u2009=\u200915.62\u00a0\u00b5g/ml) bactericidal activity towards this bacterium. Bacillus cereus ATCC 10876 was slightly less sensitive to these substances. Among them the compound 37 showed very strong activity against this bacterium. In turn, substances 35, 36, 38, 39, 41, 45, 46 and 50 had a strong activity , while compounds 40, 42, 43, 44, 47, 48 and 49\u2014good activity (Table\u00a0The compounds 35\u201350) showed also good activity with bactericidal or bacteriostatic effect against microorganisms belonging to reference staphylococci: S. aureus ATCC 25923, S. aureus ATCC 6538 and S. epidermidis ATCC 12228 . Among them, compounds 36 and 50 showed strong bactericidal activity towards S. aureus ATCC 6538 , while compound 50\u2014very strong and compounds 36 and 46\u2014strong activity towards S. epidermidis ATCC 12228 (Table\u00a0Pipemidic acid derivatives 5\u201350 show35\u201350) showed only moderate activity towards Micrococcus luteus ATCC 10240 (MIC\u2009=\u2009250\u2013500\u00a0\u00b5g/ml and MBC\u2009>\u20091000\u00a0\u00b5g/ml) and most of them except 39 and 47 against Staphylococcus aureus ATCC 43300 (MIC\u2009=\u2009250\u2013500\u00a0\u00b5g/ml and MBC\u2009=\u2009500 to >\u20091000\u00a0\u00b5g/ml). The compounds 39 and 47 indicated good (MIC\u2009=\u2009125\u00a0\u00b5g/ml and MBC\u2009>\u20091000\u00a0\u00b5g/ml) or mild (MIC\u2009=\u20091000\u00a0\u00b5g/ml and MBC\u2009>\u20091000\u00a0\u00b5g/ml) activity, respectively against S. aureus ATCC 43300 was four times better and for the compounds 35, 37, 38, 44, 45, and 46 (MIC\u2009=\u200931.25\u00a0\u00b5g/ml) was two times better against S. aureus ATCC 6538 on the basis of MIC values. The compound 50 showed two times lower MIC values (MIC\u2009=\u20097.81\u00a0\u00b5g/ml) with bactericidal effect and the compounds 36 and 46 showed two times lower MBC values (MBC\u2009=\u200931.25\u00a0\u00b5g/ml) with bactericidal effect against S. epidermidis ATCC 12228 than pipemidic acid . The MBC values for the derivative 36 towards B. subtilis ATCC 6633 (MBC\u2009=\u20093.91\u00a0\u00b5g/ml) were two times lower than for pipemidic acid (MBC\u2009=\u20097.81\u00a0\u00b5g/ml). In the case of the activity against B. cereus ATCC 10876 the MIC values for the substance 37 were two times lower (MIC\u2009=\u20097.81\u00a0\u00b5g/ml) than for pipemidic acid (MIC\u2009=\u200915.62\u00a0\u00b5g/ml) (Table\u00a035\u201350) against B. cereus ATCC 10876 is especially important due to the fact that this bacterium is responsible for an increasing number of foodborne diseases in industrial countries as well as postoperative and posttraumatic wound infections , the activity of the compounds 35\u201350) indicated also moderate or mild activity towards reference fungi belonging to yeasts. Among them the compound 42 inhibited the growth of all Candida spp. (MIC\u2009=\u2009500\u20131000\u00a0\u00b5g/ml and MFC\u2009>\u20091000\u00a0\u00b5g/ml). In turn, substances 38, 41, 43 and 49 showed mild activity towards some of them (MIC\u2009=\u20091000\u00a0\u00b5g/ml and MFC\u2009>\u20091000\u00a0\u00b5g/ml). The remaining compounds were inactive against reference Candida spp. Below is the link to the electronic supplementary material."} +{"text": "C2v symmetry and crystallizes in the chiral space group P21.The planar achiral title compound has 10H10O4, was synthesized from tetra\u00admethyl-1,4-benzo\u00adquinone. In the crystal, the almost planar mol\u00adecule (r.m.s. deviation = 0.024\u2005\u00c5) forms intra\u00admolecular hydrogen bonds between the aldehyde and hy\u00addroxy groups and exhibits C2v symmetry. This achiral mol\u00adecule crystallizes in the chiral space group P21 with inter\u00admolecular O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonding and C\u2014H\u22ef\u03c0 and C=O\u22ef\u03c0 inter\u00adactions stabilizing the crystal packing.The title compound, C Furthermore, inhibition of nucleic acid biosynthesis and of the activities of coenzyme Q mediated enzyme systems are also known for related compounds composed of 3,6-dihy\u00addroxy-4,5-di\u00admethyl\u00adbenzene-1,2-dicarbaldehyde . The C\u2014C bond lengths of the methyl substituents are 1.511\u2005(2) and 1.508\u2005(2)\u2005\u00c5, the C\u2014O bond lengths of the hy\u00addroxy substituents are 1.354\u2005(2) and 1.350\u2005(2)\u2005\u00c5, and the C\u2014C bond lengths of the aldehyde substituents are 1.464\u2005(2) and 1.462\u2005(2)\u2005\u00c5. Two intra\u00admolecular O\u2014H\u22efH hydrogen bonds between the hy\u00addroxy and aldehyde functions are observed \u2005\u00c5 (Cg1 is the centroid of ring C1\u2013C6).The C8=O2 carbonyl group is stacked on top of the aromatic ring, with the O2\u22efB\u22efCg1 (3.131\u2005\u00c5) is also found and 50% acetic acid (35\u2005ml) was added dropwise at 353\u2005K. After 10\u2005min, the reaction mixture was poured onto crushed ice (100\u2005g). The solution was filtered by vacuum filtration and a crude compound was obtained. The crude compound was dissolved in toluene and purified by silica column chromatography to afford 0.567\u2005g (yield 23.5%) of the title compound as a yellow solid . IR : 1633 (s), 3436 (m).A mixture of tetra\u00admethyl-1,4-benzo\u00adquinone and concentrated piperidine was stirred at room temperature for 35\u2005h. The mixture was evaporated and a white inter\u00admediate product was obtained. To a solution of the obtained inter\u00admediate product dissolved in acetic acid (18\u2005ml), a mixture of CrOUiso(H) = 1.5Ueq(C) for methyl H atoms, and C\u2014H = 0.95\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for the aldehyde H atoms]. O-bound H atoms were constrained using a riding model [O\u2014H = 0.84\u2005\u00c5 and Uiso(H)\u00a0= 1.5Ueq(O) for hy\u00addroxy H atoms].Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018012495/vm2211sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018012495/vm2211Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018012495/vm2211Isup3.cmlSupporting information file. DOI: 1865811CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The first of these has an unusually short N\u22efN separation of 2.616\u2005(2)\u2005\u00c5: refinement of different models against the present data set could not distinguish between a symmetrical hydrogen bond (H atom lying on the twofold axis and equidistant from the N atoms) or static or dynamic disorder models (i.e. N\u2014H\u22efN + N\u22efH\u2014N).In the crystal, two AcAP mol\u00adecules related by a crystallographic twofold axis link to H R*, 8S)-2-acetamidoocta\u00adhydro\u00adpyrrol\u00adizin-4-ium chloride\u2013N-[-hexa\u00adhydro-1H-pyrrolizin-2-yl)acetamide (1/1)], 2(C9H16N2O)\u00b7HCl or C9H17N2O+\u00b7Cl\u2212\u00b7C9H16N2O, arose as an unexpected product when 1-exo-acetamido\u00adpyrrolizidine was dissolved in CHCl3. Within the AcAP pyrrolizidine group, the unsubstituted five-membered ring is disordered over two orientations in a 0.897\u2005(5):0.103\u2005(5) ratio. Two AcAP mol\u00adecules related by a crystallographic twofold axis link to H+ and Cl\u2212 ions lying on the rotation axis, thereby forming N\u2014H\u22efN and N\u2014H\u22efCl\u22efH\u2014N hydrogen bonds. The first of these has an unusually short N\u22efN separation of 2.616\u2005(2)\u2005\u00c5: refinement of different models against the present data set could not distinguish between a symmetrical hydrogen bond (H atom lying on the twofold axis and equidistant from the N atoms) or static or dynamic disorder models (i.e. N\u2014H\u22efN + N\u22efH\u2014N). Computational studies suggest that the disorder model is slightly more stable, but the energy difference is very small.The title compound [systematic name: (1 The \u2018flap\u2019 atoms, C2 and C6, for the substituted (C1\u2013C3/N4/C8) and unsubstituted (C5\u2013C8/N4) rings, respectively, are displaced from the mean plane of the four remaining atoms (C1/C3/N4/C8 and C7/C5/N4/C8) by 0.622(1) and 0.633(2)\u2005\u00c5, respectively. The dihedral angle between these mean planes is 56.07\u2005(9)\u00b0. The acetamide group is characteristically almost planar (r.m.s. deviation = 0.0181\u2005\u00c5), but twisted from the mean plane of C1/C8/N4/C3 by a dihedral angle of 70.56\u2005(7)\u00b0. Aside from an unusually short N\u2014H\u22efN hydrogen bond, which will be discussed in subsequent sections, all geometrical parameters are within their expected ranges \u2212x\u00a0+\u00a01, y, \u2212z\u00a0+\u00a0dN\u22efN = 2.616\u2005(2)\u2005\u00c5 . Similarly, since the refined H+ position lies on the twofold axis and is equidistant from N4 and N4i, the refined N\u22efH inter\u00adatomic distance appears unusually long at 1.3080\u2005(12)\u2005\u00c5. The whole hydrogen-bonded ensemble makes an N\u22efN4i hydrogen bond, a difference-Fourier map , statically disordered or dynamically disordered . Alternative strategies for H4N inclusion in the model refined equally well , so we settled on the simplest approach, following the recommendations of F\u00e1bry . In addition to the strong N\u2014H\u22efN hydrogen bond, weaker N2\u2014H2\u2014Cl1\u2014H2i\u2014N2i inter\u00adactions link the twofold-related acetamide groups to the Cl\u2212 anion [N\u22efCl = 3.2263\u2005(11)\u2005\u00c5]. The twist of the twofold-related hydrogen-bonded pyrrolizidine moieties relative to each other, as defined by the torsion angle C8\u2014N4\u22efN4i\u2014C8i is \u221269.82\u2005(16)\u00b0. The almost planar acetamide group forms a dihedral angle with its twofold-related counterpart of 20.18\u2005(3)\u00b0. The only other intra\u00admolecular inter\u00adactions are van der Waals contacts. Estimates of the relative fractions of inter\u00admolecular contacts between individual atom types derived from a Hirshfeld-surface analysis using CrystalExplorer . The Cl\u2212 anion and the acetamide O atom each reside in pockets surrounded by hydrogen atoms, giving H\u22efCl/Cl\u22efH (16.2%) and H\u22efO/O\u22efH (12.4%), with the remainder being N\u22efH/H\u22efN and C\u22efH/H\u22efC contacts.The primary structural motif within crystals of 2AcAP\u00b7HCl consists of a pair of homochiral AcAP mol\u00adecules hydrogen bonded to H\u00c5 Table\u00a01, it is eap Fig.\u00a04 clearly \u00e1bry 2018. Nonetheet al., 2016et al., 2017A search of the Cambridge Structure Database -H-N(X)-C\u2032, where \u2018X\u2032 denotes \u2018any group\u2019, gave 45 hits. Rejection of cases where apparent close N\u22efN distances were due to disorder (four entries), and those in which the N-bound H atom was missing from the model (two entries), left 39 structures, of which three were duplicates. In the remaining 36 structures the N\u2014H\u22efN hydrogen bonds are intramol\u00adecular in 22 and intermol\u00adecular in 14. The closest N\u22efN separations occur in the intramol\u00adecular N\u2014H\u22efN hydrogen-bonded structures, the shortest being 2.419\u2005\u00c5 in EBOKOV \u2005\u00c5, although the difference is not significant, and well within the quoted precision estimate of BECHOG and the accuracy limits imposed by the spherical-atom scattering-factor approximation , and one unconstrained (B). The volume difference between these theoretical models was negligible (Avol = 1918.85\u2005\u00c53versus Bvol = 1919.63\u2005\u00c53). In the symmetric model, the N\u2014H\u22efN hydrogen atom is equidistant between the two nitro\u00adgen atoms (N\u2014H = 1.290\u2005\u00c5), whereas in model B the N\u2014H distances differ (N\u2014H = 1.194 and 1.406\u2005\u00c5). This is in agreement with the computed charge-density line profile of N\u2014H\u22efN in structure B, as shown in Fig.\u00a05B is calculated to be slightly more stable, but the energy difference (4.7\u2005meV per unit cell) is vanishingly small , both with and without symmetry constraints on charge density. In the case where symmetry constraints were absent, a small displacement (0.06\u2005\u00c5) was applied to the hydrogen atom in the N\u2014H\u22efN hydrogen bond to break symmetry in the initial geometry. The relaxations led to two structures, one with constrained twofold symmetry with Perdew\u2013Burke\u2013Ernzerhof (PBE) exchange-correlation functional calculations were carried out using the Vienna et al., 20143 in a 10\u2005ml round-bottom flask and allowing the solution to stand in a refrigerator for about a month.AcAP was synthesized and purified according to the published procedure (Pan RCH3), 0.99\u2005\u00c5 (R2CH2) and 1.00\u2005\u00c5 (R3CH). Following the advice of F\u00e1bry (2018N) was placed into difference-Fourier electron density and refined, albeit constrained to the twofold axis. An alternative model in which this H atom was allowed to ride at 50% occupancy on both N4 and N4i [symmetry code: (i) \u2212x\u00a0+\u00a01, y, \u2212z\u00a0+\u00a0N) was refined freely. Uiso(H) parameters for nitro\u00adgen-bound hydrogen atoms were refined, while for carbon-bound H atoms, Uiso(H) were set to values of either 1.2Ueq or 1.5Ueq (RCH3) of the attached atom.Crystal data, data collection, and structure refinement details are given in Table\u00a02\u00e1bry 2018, the hyd\u2212 anion were close (0.37 and 0.47\u2005\u00c5 respectively) to Cl1. Refinement of the anion as mixed Cl and Br gave an occupancy ratio of 0.934\u2005(2):0.066\u2005(2), a lower R-value (3.02%), and a flatter difference map (\u0394\u03c1 = 0.29/-0.19\u2005e\u2005\u00c5\u22123). However, the reaction included no known source of Br\u2212, so the mixed anion model was not retained.The refined displacement parameters for the ClSHELXL commands EXYZ and EADP) were used to fix overlapping fragments. Restraints were used to ensure the integrity of ill-defined or disordered groups . An alternative model using space group Cc (50:50 inversion twinned) was considered but rejected as it required hefty restraints and did not resolve the H4N atom ambiguity.To ensure satisfactory refinement for disordered groups in the structure, a combination of constraints and restraints were employed. The constraints I. DOI: 10.1107/S2056989019016517/hb7870Isup2.hklStructure factors: contains datablock(s) I. DOI: 1970639CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(6) graph-set motifs, while C\u2014H\u22efO and C\u2014H\u22efN inter\u00adactions form S(5) graph-set motifs.The title compound is a novel halogen-substituted hydrazine derivative. Intra\u00admolecular N\u2014H\u22efO inter\u00adactions form 16H14BrN3O5, is a novel halogen (Br) substituted hydrazine derivative. The hydrazine derivatives were the group of compounds with the general structure, R1R2C=NNH2 results in an extended mol\u00adecular conformation. The dihedral angle between the 5-bromo-2-meth\u00adoxy\u00adphenyl ring and the nitrophenyl ring is 4.4\u2005(3)\u00b0. Intra\u00admolecular N\u2014H\u22efO inter\u00adactions form S(6) graph-set motifs, while C\u2014H\u22efO and C\u2014H\u22efN inter\u00adactions form S(5) graph-set motifs. Symmetry-related mol\u00adecules are linked by C\u2014H\u22efO inter\u00admolecular inter\u00adactions forming an R21(10) graph-set motif. There are nearly face-to-face directional specific \u03c0\u2013\u03c0 stacking inter\u00adactions between the centroids of the nitrophenyl ring and the benzene ring of the 5-bromo-2-meth\u00adoxy group [centroid\u2013centroid distance = 3.6121\u2005(5)\u2005\u00c5 and slippage = 1.115\u2005\u00c5], which also contributes to the mol\u00adecular packing. The Hirshfeld surface analysis was performed in order to visualize, explore and qu\u00adantify the inter\u00admolecular inter\u00adactions in the crystal lattice of the title compound.The title compound, C The central bridging moiety R2C=NNHR1 adopts an all-trans conformation about the C10\u2014C9, C9\u2014N3, N3\u2014N2 and N2\u2014C5 bonds, with torsion angles of 176.0\u2005(6), \u2212178.1\u2005(5), \u2212177.0\u2005(6) and 173.6\u2005(6)\u00b0, leading to an extended mol\u00adecular conformation, thereby causing the terminal bromo\u00admeth\u00adoxy\u00adphenyl ring and nitro\u00adphenyl\u00adring to occupy almost the same plane; the dihedral angle between the rings is 4.4\u2005(3)\u00b0.Fig.\u00a01x, y, z) and the benzene ring of the 5-bromo-2-meth\u00adoxy group [centroid\u2013centroid distance = 3.6121\u2005(5)\u2005\u00c5 and slippage = 1.115\u2005\u00c5], which also contributes to the mol\u00adecular packing. The Br atom does not take part in any inter\u00adactions. The nearest Br\u22efC7, which are summarized in Table\u00a02et al., 2013et al., 2011Hirshfeld surface analysis serves as a powerful tool for gaining additional insight into inter\u00admolecular inter\u00adactions of mol\u00adecular crystals. The Hirshfeld surfaces are mapped with 2D fingerprint plots presented using et al., 2016et al., 2011et al. 2010et al. 2010et al., 2015et al., 1985et al., 2003A and B), 9.30\u2005(6)\u00b0, and 13.01\u2005(10) and 14.05\u2005(10)\u00b0 (in mol\u00adecules A and B), respectively, compared to 4.4\u2005(3)\u00b0 in the title compound. The crystal packing of the two compounds is significantly different. In AYSOD, N\u2014H groups do not form hydrogen bonds, in DUSBID, the mol\u00adecules are linked by N\u2014H\u22ef\u03c0 inter\u00adactions, and in DUSNUB, both mol\u00adecules form inversion dimers linked by pairs of N\u2014H\u22efO hydrogen bonds, thereby generating et al., 1995et al., 2013While searching for 2-phenyl\u00adhydrazine in the Cambridge Structural Database with a slight excess of 5-bromo-2-meth\u00adoxy\u00adbenzaldehyde (0.215\u2005mg) in an acetic acid solution (10\u2005ml). The reaction mixture was refluxed for 8\u2005h. The solid product formed during reflux was filtered off, washed and dried over anhydrous calcium chloride in a vacuum desiccator (yield 75%). The final product was soluble in acetone, dimethyl sulfoxide (DMSO), di\u00admethyl\u00adformamide (DMF), methanol, ethanol and ethyl acetate, Uiso(H) = xUeq(C), where x = 1.5 for methyl and x = 1.2 for all other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018011325/dx2007sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018011325/dx2007Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018011325/dx2007Isup3.cmlSupporting information file. DOI: 1860856CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S),18(R)\u2010EpETE and 17(R),18(S)\u2010EpETE. In this study, we demonstrated stereoselective differences of 17(S),18(R)\u2010EpETE and 17(R),18(S)\u2010EpETE in amelioration of skin contact hypersensitivity and found that anti\u2010inflammatory activity was detected in 17(S),18(R)\u2010EpETE, but not in 17(R),18(S)\u2010EpETE. In addition, we found that cytochrome P450 BM\u20103 derived from Bacillus megaterium stereoselectively converts EPA into 17(S),18(R)\u2010EpETE, which effectively inhibited the development of skin contact hypersensitivity by inhibiting neutrophil migration in a G protein\u2010coupled receptor 40\u2010dependent manner. These results suggest the new availability of a bacterial enzyme to produce a beneficial lipid mediator, 17(S),18(R)\u2010EpETE, in a stereoselective manner. Our findings highlight that bacterial enzymatic conversion of fatty acid is a promising strategy for mass production of bioactive lipid metabolites.Dietary intake of \u03c93 polyunsaturated fatty acids such as eicosapentaenoic acid and docosahexaenoic acid is beneficial for health control. We recently identified 17,18\u2010epoxyeicosatetraenoic acid as a lipid metabolite endogenously generated from eicosapentaenoic acid that exhibits potent anti\u2010allergic and anti\u2010inflammatory properties. However, chemically synthesized 17,18\u2010EpETE is enantiomeric due to its epoxy group\u201417( One enantiomer may be responsible for the therapeutic effects, whereas the other enantiomer can be inactive or exert undesired effects. For example, 2(S)\u2010hydroxy oleic acid induces greater reduction in the tumor volume of lung cancer than does 2(R)\u2010hydroxy oleic acid.R),18(S)\u2010EpETE\u2014but not 17(S),18(R)\u2010EpETE\u2014has vasodilatory effects on arteries, again suggesting that chirality is important for determination of biological activity.The stereochemistry of oxidized fatty acids is a critical determinant of their biological activity. Indeed, 17,18\u2010EpETE is enantiomeric due to its epoxy groups\u201417(R)\u2010HEPE which plays important roles as a SPM and a precursor of resolvin E1 (RvE1) was shown to be generated by hydroxylation of EPA using Bacillus megaterium homogenates.B.\u00a0megaterium, catalyzed the epoxidation of unsaturated fatty acids and converted EPA into 17(S),18(R)\u2010EpETE in a stereoselective manner,Although 17,18\u2010EpETE is endogenously generated from EPA by cytochrome P450 (CYP), microorganisms also harbor various types of CYPs which catalyze a wide range of reactions including epoxidation and hydroxylation.Contact hypersensitivity is a commonly used mouse model of human allergic contact dermatitis, which includes sensitization and elicitation phases. In the sensitization phase, dendritic cells (DCs) migrate to the draining lymph nodes and activate T cells for the induction of memory\u2010type T cells. In the elicitation phase, on exposure to the same contact allergen as experienced during the sensitization phase, neutrophils and memory\u2010type T cells infiltrate the inflamed skin, where they produce pro\u2010inflammatory cytokines that lead to the development of skin swelling, rashes, and edema.S),18(R)\u2010EpETE produced by BM\u20103 (BM\u20103 17(S),18(R)\u2010EpETE) ameliorated CHS by inhibiting neutrophil migration into inflamed skin in a GPR40\u2010dependent manner. These findings reveal the activity of the 17(S),18(R)\u2010EpETE enantiomer and the availability of a bacterial enzyme that produces this bioactive lipid metabolite in a stereoselective manner.In this study, we found that 17(22.1Wild\u2010type (WT) C57BL/6J female mice were purchased from SLC and kept in a specific pathogen\u2010free animal facility at the National Institutes of Biomedical Innovation, Health and Nutrition (NIBIOHN) for at least 1\u00a0week before use in experiments. GPR40\u2010deficient mice have been described previously2.2S),18(R)\u2010EpETE and 17(R),18(S)\u2010EpETE ((\u00b1)17,18\u2010EpETE), a commercially available Cayman (\u00b1)17,18\u2010EpETE . In some experiments, mice were treated with stereoselective 17(S),18(R)\u2010EpETE (>99% enantiomeric excess) or 17(R),18(S)\u2010EpETE (>99% enantiomeric excess), which were purified from synthesized (\u00b1)17,18\u2010EpETE, or BM\u20103 17(S),18(R)\u2010EpETE. These lipids were injected intraperitoneally into mice by 100\u00a0ng/animal at 30\u00a0minutes before DNFB treatment. In some experiments, BM\u20103 17(S),18(R)\u2010EpETE were injected intraperitoneally into mice by 1\u00a0\u00b5g, 100\u00a0ng, or 10\u00a0ng/animal in order to evaluate dose response. We used 0.5% (vol/vol) ethanol dissolved in PBS as a vehicle control.Contact hypersensitivity was induced as described previously.2.3The (\u00b1)17,18\u2010EpETE was prepared from EPA as described previously with some modifications.For purification of stereoselective 17,18\u2010EpETE, HPLC was performed using a CHIRALCEL OJ\u2010RH packed column . The isocratic mobile phase was a mixture of methanol (Wako) and water containing 0.1% (vol/vol) formic acid , which was pumped at a flow rate of 1.2\u00a0mL/min. The column was maintained at 30\u00b0C, and the eluent was monitored at a wavelength of 205\u00a0mm. As a control, commercially available Cayman (\u00b1)17,18\u2010EpETE was used for HPLC analysis.2.4E.\u00a0coli DH5\u03b1 (Nippon Gene), and transformed by heat shock at 42\u00b0C for 30\u00a0seconds. After the transformation, E. coli DH5\u03b1 was inoculated to modified LB agar medium containing 25\u00a0\u03bcg/mL kanamycin sulfate (Wako) and cultured at 37\u00b0C for 18\u00a0hours. Modified LB agar medium was comprised of 1% (wt/vol) Difco select soytone , 0.5% (wt/vol) Bacto yeast extract , 1% (wt/vol) sodium chloride (Wako) and 1.5% (wt/vol) agar powder (Wako). The resultant colonies were picked and inoculated to modified LB liquid culture medium containing 25\u00a0\u03bcg/mL kanamycin sulfate and cultured at 37\u00b0C for 18\u00a0hours. Modified LB liquid culture medium was comprised of 1% (wt/vol) Difco select soytone, 0.5% (wt/vol) Bacto yeast extract and 1% (wt/vol) sodium chloride. The plasmid DNA was extracted using QIAprep Spin Miniprep Kit (Qiagen) according to the manufacturer's instructions. In this way, pFusionF87V\u2010Km plasmid was constructed.BM3\u2010encoding pFusionF87V plasmid was ligated into pET\u201024d (+) (Merck) at KNC Laboratories .E.\u00a0coli DH5\u03b1 and transformed by heat shock at 42\u00b0C for 30\u00a0seconds. The recombinant E.\u00a0coli was inoculated to modified LB agar medium containing 25\u00a0\u03bcg/mL kanamycin sulfate\u00a0and cultured at 37\u00b0C for 18\u00a0hours. The resultant colonies were inoculated to modified LB liquid culture medium containing 25\u00a0\u00b5g/mL kanamycin sulfate and cultured at 37\u00b0C for 18\u00a0hours. The plasmid DNA was extracted using QIAprep Spin Miniprep Kit. In this way, pFusionBM3\u2010WT plasmid which has no mutation was constructed.We next used pFusionF87V\u2010Km plasmid as a template for construction of pFusionBM3\u2010WT plasmid to give rise to large amount of 17,18\u2010EpETE. The primers for inverse PCR were as follows: primer 1, 5'\u2010TTTACAAGCTGGACGCATGA\u20103' and primer 2, 5'\u2010TAACCCGTCTCCTGCAAAATCAC\u20103'. The fragments were self\u2010ligated using Ligation\u2010Convenience kit after phosphorylation of 5' ends by T4 polynucleotide kinase (Takara Bio). The resultant solution was suspended with ECOS Competent E.\u00a0coli BL21 (DE3) which is expression host strain, the pFusionBM3\u2010WT was suspended with ECOS Competent E.\u00a0coli BL21 (DE3) (Nippon Gene) and transformed by heat shock at 42\u00b0C for 30\u00a0seconds. After the transformation, E.\u00a0coli BL21 (DE3) was inoculated to modified LB agar medium containing 25\u00a0\u03bcg/mL kanamycin sulfate and cultured at 37\u00b0C for 18\u00a0hours. The resultant colonies were inoculated to modified LB liquid culture medium containing 25\u00a0\u00b5g/mL kanamycin sulfate and cultured at 28\u00b0C for 18\u00a0hours. The glycerol stock of pFusionBM3\u2010WT/BL21 (DE3) was made by mixing cultured solution and 50% (vol/vol) glycerol (Wako) in 2:1 (vol/vol) and stored at\u00a0\u221220\u00b0C.In order to carry pFusionBM3\u2010WT into 2.5Cultivation was performed at KNC Laboratories. For preculture, 100\u00a0\u00b5L of pFusionBM3\u2010WT/BL21 (DE3) glycerol stock was inoculated in 500\u00a0mL of modified LB liquid culture medium containing 25\u00a0\u00b5g/mL kanamycin sulfate and cultured at 25\u00b0C for 22\u00a0hours with shaking at 120\u00a0rpm.D\u2010thiogalactopyranoside and cultured at 20\u00b0C for 47\u00a0hours with ventilation rate of 75\u00a0L/min. 2\u00a0\u00d7\u00a0YT medium was comprised of 1.6% (wt/vol) Difco select soytone, 1% (wt/vol) Bacto yeast extract and 0.5% (wt/vol) sodium chloride. pH in culture was maintained at pH 7.0\u00a0\u00b1\u00a00.1 using 25% (vol/vol) ammonia solution (Wako) and 2\u00a0mol/L phosphoric acid (Wako), and dissolved oxygen was maintained at DO 1.5\u00a0\u00b1\u00a00.5\u00a0ppm by stirring.1 L of precultured liquid was added to 150 L of modified 2\u00a0\u00d7\u00a0YT medium containing 25\u00a0\u00b5g/mL kanamycin sulfate, 80\u00a0\u00b5g/mL 5\u2010aminolevulinic acid (Wako), 100\u00a0\u00b5M ammonium iron (II) sulfate hexahydrate (Wako), 250\u00a0\u00b5mol/L isopropyl \u03b2\u20102.6Bioconversion of EPA into 17,18\u2010EpETE was performed at KNC Laboratories. 1.5 L of 1\u00a0mol/L EPA was added to cultured medium and incubated at 20\u00b0C for 71.5\u00a0hours with ventilation rate of 20\u00a0L/min in order to convert EPA into 17,18\u2010EpETE. pH was maintained at pH 7.0\u00a0\u00b1\u00a00.1 using 25% (vol/vol) ammonia solution and 2\u00a0mol/L phosphoric acid, and dissolved oxygen was maintained at DO 1.5\u00a0\u00b1\u00a00.5\u00a0ppm by stirring.In order to stop reaction and kill bacteria, 35\u00a0L of ethanol (Wako) was added to reaction mixture and cultured at 20\u00b0C for 46\u00a0hours with ventilation rate of 20\u00a0L/min. pH was maintained at pH 7.0\u00a0\u00b1\u00a00.1 using 25% (vol/vol) ammonia solution and 2\u00a0mol/L phosphoric acid, and dissolved oxygen was maintained at DO 1.5\u00a0\u00b1\u00a00.5\u00a0ppm by stirring. In order to confirm whether the bacteria are dead, the reaction liquid was inoculated to modified LB agar medium and cultured at 37\u00b0C for 18\u00a0hours, then we confirmed that the colonies were not formed.2.7S),18(R)\u2010EpETE was performed at KNC Laboratories. Fifteen kilograms of Diaion HP20 were added to 150 L of EPA reaction solution (20% (vol/vol) ethanol aqueous solution) and stirred for 1\u00a0hour to adsorb 17,18\u2010EpETE, which was confirmed by the analysis of the supernatant with HPLC Prominence System (Shimadzu). The condition of HPLC analysis is as follows: flow rate; 1.0\u00a0mL/min, column temperature; 40\u00b0C, UV wavelength; 205\u00a0nm, injection volume; 10\u00a0\u00b5L, column; C18 column , mobile phase; (A) 0.05% (vol/vol) formic acid aqueous solution and (B) acetonitrile. The eluent gradients were 45%\u201055% (vol/vol) B for 0\u201012\u00a0minutes, 55%\u201075% (vol/vol) B for 12\u201019\u00a0minutes, 75%\u2010100% (vol/vol) B for 19\u201020\u00a0minutes, 100% (vol/vol) B for 20\u201028\u00a0minutes, 100%\u201045% (vol/vol) B for 28\u201029\u00a0minutes, and 45% (vol/vol) B for 29\u201040\u00a0minutes. After the absorption of 17,18\u2010EpETE, HP20 was removed from the EPA reaction solution, and washed four times with purified water. Then, HP20 was soaked in 20% (vol/vol) ethanol aqueous solution and stored at 4\u00b0C.Purification of BM\u20103 17 ethanol aqueous solution containing 0.05% (vol/vol) acetic acid). This solution was charged to Inertsil ODS\u20103 column that had been equilibrated with 20% (vol/vol) ethanol aqueous solution containing 0.05% (vol/vol) acetic acid at 50\u00a0mL/min flow rate. After charged, the column was washed with 20% (vol/vol) ethanol aqueous solution containing 0.05% (vol/vol) acetic acid, and eluted with 70% (vol/vol) ethanol aqueous solution containing 0.05% (vol/vol) acetic acid at 45\u00a0mL/min flow rate. The elution sample containing 17,18\u2010EpETE was stored at \u221280\u00b0C as a low purity sample .In order to obtain high purity sample, the low purity sample was charged to Inertsil ODS\u20103 column that had been equilibrated with 20% (vol/vol) ethanol aqueous solution containing 0.05% (vol/vol) acetic acid at 15\u00a0mL/min flow rate. After charged, the column was washed with 20% (vol/vol) ethanol aqueous solution containing 0.05% (vol/vol) acetic acid, and eluted with 55% (vol/vol) ethanol aqueous solution containing 0.05% (vol/vol) acetic acid at 10\u00a0mL/min flow rate. The elution sample containing 17,18\u2010EpETE was stored at \u221280\u00b0C as a high purity sample .S),18(R)\u2010EpETE for biological assay.In order to replace the solvent to ethanol, the elution sample containing 17,18\u2010EpETE was diluted with Milli\u2010Q water containing acetic acid ethanol aqueous solution containing 0.05% (vol/vol) acetic acid). The resultant solution was charged to Inertsil ODS\u20103 column that had been equilibrated with 20% (vol/vol) ethanol aqueous solution containing 0.05% (vol/vol) acetic acid at 15\u00a0mL/min flow rate. After charged, the column was washed with 20% (vol/vol) ethanol aqueous solution containing 0.05% (vol/vol) acetic acid, followed by washed with 20% (vol/vol) ethanol aqueous solution, and eluted with 100% (vol/vol) ethanol at 15\u00a0mL/min flow rate. The elution sample containing 17,18\u2010EpETE was stored at \u221280\u00b0C as a very high purity sample , and used as BM\u20103 17 staining, frozen tissue sections were stained in hematoxylin solution (Wako) for 10\u00a0minutes and washed with water for 30\u00a0minutes. Then, the sections were stained in 1% eosin Y solution (Wako) for 1\u00a0minute and dehydrated through increasing concentrations of ethanol . Finally, stained tissue sections were dehydrated in xylene for 3\u00a0minutes and mounted in Permount .For immunohistologic analysis, frozen tissue sections were washed with PBS for 10\u00a0minutes and then blocked in 2% (vol/vol) newborn calf serum (Equitech\u2010Bio) in PBS for 30\u00a0minutes at room temperature in an incubation chamber (Cosmo Bio). Tissue sections were then incubated in an incubation chamber overnight at 4\u00b0C with fluorescein isothiocyanate\u2013anti\u2010Ly6G monoclonal antibody in 2% (vol/vol) newborn calf serum in PBS. Then, samples were washed once for 5\u00a0minutes in 0.1% (vol/vol) Tween\u201020 in PBS and then in PBS only for 5\u00a0minutes. To visualize nuclei, tissue sections were stained with 4\u02b9,6\u2010diamidino\u20102\u2010phenylindole for 10\u00a0minutes at room temperature in the incubation chamber. Finally, tissue sections were washed twice with PBS for 5\u00a0minutes each, and mounted in Fluoromount (Diagnostic BioSystems) and examined under a fluorescence microscope .2.9The isolation of cells from ear tissue and their flow cytometric analysis were performed as described previously.Cell suspensions were filtered using a cell strainer and cells counted. Cells were stained using an anti\u2010CD16/32 monoclonal antibody to avoid nonspecific staining, and dead cells were stained with 7\u2010aminoactinomycin D . The cells were further stained with the following antibodies: fluorescein isothiocyanate\u2013anti\u2010Ly6G , allophycocyanin\u2013Cy7\u2013anti\u2010CD11b , and BV421\u2013anti\u2010CD45 . Samples were analyzed using MACSQuant . Data were analyzed using FlowJo 9.9 software (TreeStar).2.105 cells) were suspended in HBSS containing 0.2% bovine serum albumin (Sigma Aldrich) and allowed to adhere to fibronectin\u2010coated coverslips (Neuvitro) for 15\u00a0minutes at 37\u00b0C in a 5% CO2 incubator. Neutrophils were treated with either 1000, 100, 10, or 1\u00a0nmol/L of commercially available Cayman (\u00b1)17,18\u2010EpETE, BM\u20103 17(S),18(R)\u2010EpETE, 18\u2010HEPE , RvE1 , or 0.03% (vol/vol) ethanol (vehicle control) for 15\u00a0minutes and then stimulated with 1\u00a0\u03bcmol/L N\u2010formyl\u2010methionyl\u2010phenylalanine or 100\u00a0nmol/L leukotriene B4 for 2\u00a0minutes at 37\u00b0C in a 5% CO2 incubator. Neutrophils were fixed in 4% paraformaldehyde , permeabilized using 0.5% (vol/vol) Triton X\u2010100 in PBS, and stained with 100\u00a0nmol/L Acti\u2010stain 488\u2013phalloidin (Cytoskeleton) for 30\u00a0minutes at room temperature. Finally, cell nuclei were stained by incubating neutrophils with 4\u02b9,6\u2010diamidino\u20102\u2010phenylindole for 30\u00a0seconds at room temperature. Images were obtained with Leica TCS SP8 confocal microscopy (Leica Microsystems).Neutrophils were purified from bone marrow as described previously.2.11Actin\u03b2 sense, 5'\u2010aaggccaaccgtgaaaagat\u20103'; Actin\u03b2 antisense, 5'\u2010gtggtacgaccagaggcatac\u20103'; interferon\u2010\u03b3 (Ifn\u2010\u03b3) sense, 5'\u2010atctggaggaactggcaaaa\u20103'; Ifn\u2010\u03b3 antisense, 5'\u2010ttcaagacttcaaagagtctgaggta\u20103'; interleukin (Il)\u201017 sense, 5'\u2010cagggagagcttcatctgtgt\u20103'; Il\u201017 antisense, 5'\u2010gctgagctttgagggatgat\u20103'; Cxcl1 sense, 5'\u2010gactccagccacactccaac\u20103'; Cxcl1 antisense, 5'\u2010tgacagcgcagctcattg\u20103'; Cxcl2 sense, 5'\u2010aaaatcatccaaaagatactgaacaa\u20103'; Cxcl2 antisense, 5'\u2010ctttggttcttccgttgagg\u20103'; Cxcl9 sense, 5'\u2010cttttcctcttgggcatcat\u20103'; Cxcl9 antisense, 5'\u2010gcatcgtgcattccttatca\u20103'; Cxcl10 sense, 5'\u2010gctgccgtcattttctgc\u20103'; Cxcl10 antisense, 5'\u2010tctcactggcccgtcatc\u20103'.Reverse transcription and quantitative PCR analysis were performed as described previously.2.12P\u00a0<\u00a00.05 was considered significant.Statistical significance was evaluated through one\u2010way ANOVA using Prism 3.03 software (GraphPad Software). A 33.1S),18(R)\u2010EpETE from 17(R),18(S)\u2010EpETE. In this issue, we first chemically synthesized a large amount of (\u00b1)17,18\u2010EpETE from EPA and found that, like commercially available Cayman (\u00b1)17,18\u2010EpETE, synthesized (\u00b1)17,18\u2010EpETE showed 2 peaks using HPLC system with chiral column. The bacterial enzyme BM\u20103 has been reported to stereoselectively convert EPA into 17(S),18(R)\u2010EpETE.S),18(R)\u2010EpETE was identical with the second peak of (\u00b1)17,18\u2010EpETE ,18(S)\u2010EpETE.We previously reported that commercially available Cayman (\u00b1)17,18\u2010EpETE showed potent anti\u2010allergic and anti\u2010inflammatory properties.3.2S),18(R)\u2010EpETE showed anti\u2010inflammatory activity in inhibiting ear swelling but 17(R),18(S)\u2010EpETE had little effect 17,18\u2010EpETE, 17,18(R)\u2010EpETE exerted anti\u2010inflammatory activity, we next examined whether 17(S),18(R)\u2010EpETE stereoselectively produced by BM\u20103 suppresses CHS. We found that BM\u20103 17(S),18(R)\u2010EpETE suppressed ear swelling ,18(R)\u2010EpETE decreased inflammation in the skin ,18(R)\u2010EpETE decreased the number of neutrophils in inflamed ears ,18(R)\u2010EpETE ,18(R)\u2010EpETE in CHS by evaluating ear swelling and neutrophil numbers. Both ear swelling and neutrophil numbers were decreased at the dose of 1\u00a0\u00b5g and 100\u00a0ng/animal, but the anti\u2010inflammatory effects were hardly observed at the dose of 10\u00a0ng/animal ,18(R)\u2010EpETE was shown to be effective at the dose more than 100\u00a0ng/animal.Given that 17,18(R)\u2010EpETE on pseudopod formation in neutrophils isolated from bone barrow. We found that fMLP\u2010induced pseudopod formation was inhibited by treatment with BM\u20103 17(S),18(R)\u2010EpETE ,18(R)\u2010EpETE was absent when the neutrophils were prepared from GPR40\u2010deficient mice. We also found that BM\u20103 17(S),18(R)\u2010EpETE inhibited LTB4\u2010induced pseudopod formation in a GPR40\u2010dependent manner ,18(R)\u2010EpETE treatment ,18(R)\u2010EpETE did not inhibit the expression of Ifn\u2010\u03b3 ,18(R)\u2010EpETE acts on neutrophil selectively.We next evaluated the inhibitory effect of BM\u20103 17,18(R)\u2010EpETE with EPA\u2010derived fatty acid metabolites, RvE1 and 18\u2010HEPE, for inhibition of pseudopod formation ,18(R)\u2010EpETE showed inhibitory effects at the dose of 10\u00a0nmol/L, while RvE1 and 18\u2010HEPE lost their activities. BM\u20103 17(S),18(R)\u2010EpETE also lost its activity at the dose of 1\u00a0nmol/L.We compared the effectiveness of BM\u20103 17,18(R)\u2010EpETE at the dose of 10\u00a0nmol/L, while RvE1 and 18\u2010HEPE showed inhibitory effects. Both RvE1 and 18\u2010HEPE lost its activity at the dose of 1\u00a0nmol/L. These results indicated that EPA\u2010derived bioactive lipid mediators of BM\u20103 17(S),18(R)\u2010EpETE, RvE1, and 18\u2010HEPE all possess anti\u2010inflammatory activity by inhibiting neutrophil pseudopod formation.In the case of LTB44, which is secreted by neutrophils, promotes secondary neutrophil migration.S),18(R)\u2010EpETE inhibited both fMLP\u2010 and LTB4\u2010induced pseudopod formation, and RvE1 and 18\u2010HEPE also inhibited both fMLP\u2010 and LTB4\u2010induced pseudopod formation. Consistent with these findings, a previous report showed that RvE1 and 18\u2010HEPE reduced neutrophil transmigration.4 receptor, BLT1.4\u2010induced pseudopod formation through BLT1 antagonistic activity in DC.S),18(R)\u2010EpETE inhibited neutrophil pseudopod formation by GPR40\u2010mediated pathway, while 18\u2010HEPE and RvE1 used ChemR23 and/or BLT1 as functional receptors.Actin polymerization and neutrophil migration are induced by different kinds of chemoattractants. The fMLP promotes primary neutrophil migration in response to bacterial infection, while LTBS)\u2010 and (R)\u2010thalidomide was introduced as a sedative medicine in the late 1950s, but it was withdrawn due to teratogenicity of (S)\u2010thalidomide, indicating the importance of stereoselective production of candidate medicines.S)\u2010 and (R)\u2010enantiomers of salbutamol exert different effects: the (R)\u2010enantiomer of salbutamol binds the B2\u2010adrenergic receptor with greater affinity than the (S)\u2010enantiomer and is responsible for salbutamol's bronchodilation activity.The use of single\u2010enantiomer medicines can potentially lead to simpler and more selective pharmacologic profiles, because the enantiomers of a chiral compound may differ significantly in their bioavailability.S),18(R)\u2010EpETE stereoselectively showed anti\u2010inflammatory activity in CHS. Conversely, it is reported that 17(R),18(S)\u2010EpETE\u2014but not 17(S),18(R)\u2010EpETE\u2014is a potent vasodilator and stimulates calcium\u2010activated potassium channels, which lead to the relaxation of rat cerebral artery vascular smooth muscle cells.S),18(R)\u2010EpETE\u2013GPR40 axis suppresses CHS, whereas the 17(R),18(S)\u2010EpETE\u2013calcium\u2010activated potassium\u2010channel axis achieves arterial relaxation.S),18(R)\u2010EpETE as a single\u2010enantiomer therapy might decrease the risk of side effects due to vasodilatory activity, such as a rapid decrease in blood pressure and increased skin redness.In this study, we found that BM\u20103 17(R),18(S)\u2010EpETE, whereas Cyp4f18 in mice and CYP2D6 in humans\u2014like BM\u20103\u2014selectively generate 17(S),18(R)\u2010EpETE.R),18(S)\u2010EpETE and 17(S),18(R)\u2010EpETE.We previously found that dietary linseed oil, which contains large amounts of \u03b1\u2010linolenic acid, a precursor of EPA and DHA, increases the amount of \u03c93 PUFA\u2010derived metabolites in the body.S),18(R)\u2010EpETE might reflect differences in CYP activity or expression level. For example, the expression level of Cyp1a2 is upregulated by the ligand\u2010activated transcription factor aromatic hydrocarbon receptor (AhR).R),18(S)\u2010EpETE might be modulated through diet.S),18(R)\u2010EpETE and 17(R),18(S)\u2010EpETE.CYPs harbor gene polymorphisms, which cause different enzymatic activities among individuals.Bacillus bacteria, including B.\u00a0megaterium, are used for the production of fermented foods, such as the soybean products natto and miso. Therefore, the production level of lipid metabolites likely is affected not only by the enzymes in the body but also by enzymes derived from microorganisms in fermented food, suggesting that the production levels of 17(S),18(R)\u2010EpETE could be increased by eating fermented foods containing Bacillus bacteria.B.\u00a0megaterium homogenate reportedly yields 17,18\u2010EpETE and 18\u2010HEPE, whereas BM\u20103 primarily catalyzes epoxidation and produces 17,18\u2010EpETE from EPA, thus suggesting that not only BM\u20103, various types of CYPs in B.\u00a0megaterium contribute to the production of 17,18\u2010EpETE and 18\u2010HEPE.Because microorganisms metabolize fatty acids, microorganisms can affect the lipid profile. In particular, microorganisms are used for the production of fermented foods, which may often contain abundant amounts of lipid metabolites. For example, S),18(R)\u2010EpETE can reduce CHS. The 17(S),18(R)\u2010EpETE\u2013GPR40 axis played a key role in the amelioration of CHS by inhibiting neutrophil migration. These results suggest that bacterial fermentation with BM\u20103 activity is a promising tool for the stereoselective mass\u2010production of 17(S),18(R)\u2010EpETE.In conclusion, the present study showed enzymatically produced 17(There are no conflict of interest to declare. AS, TN, and JK designed the research and wrote the paper. AS, TN, SK, SP, NM, MS, SM, EN, and JO performed experiments, analyzed data, and discussed the results. TH, PT, SH, KH, and KK provided technical help and discussed the results.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file."} +{"text": "Recently oxidative stress induced maladies have amplified owing to sedentary lifestyle and monotonous diet. Introduction of plant based biomolecules may be a suitable strategy to cope with the lipid peroxidation. In this context, black tea polyphenols (theaflavin & thearubigins) are in fame among the scientific community as cost effective therapeutic agents owing to their safety, economics, structural diversity and ability to modulate various lipid peroxidation responses by halting the expression of different metabolic targets.0 (control diet without supplementation), T1 , T2 & T3 for the period of 56\u00a0days. Alongside, a control study was also carried out for comparison by involving normal rats fed on arginine free diet. The body weight, lipid profile, glycemic responses, Renal function test, liver function test, antioxidant indices and hematological parameters were estimated at the termination of study.The mandate of present investigation was to first time check the synergism among the isolated theaflavins & thearubigins against lipid peroxidative indicators both in vitro and in vivo. Purposely, theaflavins and thearubigins were isolated from black tea through solvent partition methods by using different solvents and time intervals and subjected to in vitro characterization through different antioxidant indices to access the in vitro lipid peroxidation shooting effect of these bioactive moieties. Moreover, individual theaflavins contents also estimate through HPLC. For evaluation of in vivo antioxidant effect, renal malfunction was induced through arginine and forty rats were divided in four groups and 04 types of diets were given i.e. TThe results indicated that theaflavins and thearubigins isolation was significantly affected by time of extraction and solvent. In this context, aqueous ethanol at 60\u00a0min extraction interval caused maximum extraction. Likewise, theaflavins isolate exhibited more antioxidant activity as compared to thearubigins. Moreover, the theaflavins and thearubigins based experimental diets imparted significant reduction in Lipid profile, glucose content, renal function tests and TBARS with enhancement in insulin, HDL and hematological parameters. In this context, theaflavin based diet caused maximum reduction in lipid profile and TBARS better as compared to thearubigins and theaflavins + thearubigins based. However, theaflavin+ thearubigins based diet caused highest glucose, urea & creatinine decline and maximum insulin increase & antioxidant indices as compared to other nutraceuticals.It was deduced that theaflavins & thearubigins have strong antioxidative potential both in in vitro as well as in vivo to tackle the menace associated with lipid peroxidation. Likewise, preventive role of theaflavin in lipid peroxidation is mainly attributed to its ability to cease the chain reaction. Apart from free radical scavenging and metal chelating abilities, theaflavin has potential to activate certain antioxidant enzymes like glutathione-S-transferase (GST), glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (CAT) thereby reduces lipid peroxidation. Likewise, thearubigins or polymeric black tea polyphenols (PBPs) are the oxidative products of phenolics and their production accelerated after 75% conversion of catechins into flavan-3-ol molecules. They have ability to activate phase II enzymes by inducing transcriptional upregulation in lung and liver [Globally, therapeutic worth of tea has been established against numerous maladies. The health escalating perspectives of tea has been attributed to his distinguished polyphenols. Accordingly, theaflavins and thearubigins are the promising polyphenols belong to subclass flavanolos. They produced when fresh tea leaves undergo enzymatic fermentation triggered by polyphenol oxidase and catalase resulted in structural variations in catechins thus formed theaflavin and thearubigins \u20133. Strucnd liver .Chronic renal failure (CRF) is an irreversible loss to functioning nephrons resulting numerous disorders of blood vessels, glomeruli, tubules and renal interstitium . VariousThe mandate of current investigation to isolate the theafavins and thearubigins from black tea Qi-men variety by using different solvents and time intervals, in vitro characterization and preparation of nutraceutical intervention against renal malfunctioning in arginine induced liver damage rats.Black tea variety (Qi-Men) was procured from the National Tea Research Institute (NTRI), Shinkiari, Mansehra. The reagents and standards were purchased from Merck and Sigma-Aldrich . For efficacy trial, Male Sprague Dawley rats were housed in the Animal Room of Physiology department of GCUF. For biological assay, diagnostic kits were purchased from Sigma-Aldrich, Bioassay and Cayman Chemicals .Theaflavin and thearubigins were extracted by using water, methanol and ethanol at 30, 60 and 90\u00a0min intervals and isolated by solvent partition method . Initial18 column , 10\u00a0\u03bcL sample through auto sampler (WISP Model 710) and column temperature 40\u00a0\u00b0C. The composition of mobile phase was acetonitrile, ethylacetate and 0.05% phosphoric acid in ratio of 21:3:76 with flow rate of 1\u00a0mL/min and using on UV/vis detector (model 481) and measurement wavelength was 278\u00a0nm.The isolated samples of theaflavins were characterized for their fractions through HPLC . The conditions for HPLC were CAntioxidant capacity of isolated theaflavin and thearubigins fractions were determined through antioxidant indices including total antioxidant activity, free radical scavenging activity and ferric reducing antioxidant power as following methods. For this purpose, the isolated fractions both from theaflavin and thearubigins were mixed in water, methanol and ethanol (1\u00a0mg/mL) to be further utilized in antioxidant indices estimation.V/vis Spectrophotometer (CECIL CE7200) against control and expressed results as mg gallic acid/100\u00a0g.Total phenolics of resultant isolates were estimated spectrophotometricaly using Folin-Ciocalteau method . Briefly\u03b2-carotene and linoleic acid assay was applied to measure the total antioxidant capacity . PurposeDPPH radical scavenging activity was measured according the procedure of . InitialThe FRAP test was performed according to the guidelines of . 0.5\u00a0mL Effect of isolated theaflavin and thearubigins fraction on glucose diffusion was assessed using glucose oxidase kit and 1.65\u00a0mM D-glucose solution .For biological assay, rats were divided in to four homogeneous groups fed on experimental diet. The common experimental diet was formulated using corn oil (10%), protein (10%), corn starch (64%), cellulose (10%), mineral (3%) vitamin mixture (1%) alongside arginine @ 2% for the induction of renal malfunctioning. For comparison a control study was also conducted by providing the normal diet (same composition except for arginine). However, common experimental diet from both studies further divided into four groups on the bases of addition of active ingredients, Diet 1 , Diet 2, (theaflavins supplementation @ 1\u00a0g), Diet 3, (Thearubigins supplementation @ 1\u00a0g) and Diet 4 . Afterwards All the ingredients were mixed then oven baked for 10\u00a0min. The dose was selected by carried out a 21\u00a0days safety trial (Data not included).n\u2009=\u20096 rats/dose). The treatments were given in both acute (single dose followed by a 48-h observation period) and sub-acute . The hematological analysis as well as general physical examination was carried out [In safety trial, rats were provided all the active ingredients orally @ of 250, 500, 1000, 1500, 2000 and 3000\u00a0mg/kg-bw and relative humidity (55\u2009\u00b1\u20095%) along with 12\u00a0h light-dark period. At the initiation of study, some rats were sacrificed to establish the baseline trend. For the induction of renal malfunctioning initially high arginine diet @ 2% was administrated for a period of 7 days. During that tenure urea and creatinine levels were observed to estimate the onset of renal malfunctioning. Afterwards when values of both test deviate 25% from normal then the original study was started. The study comprised of four groups of rats ten in each . Accordingly, four types of experimental diets were given i.e. T0 , T1, T2 and T3 by protocols of , high deGlucose concentration was estimated by GOD-PAP method by following the protocol of , howeverThe serum samples were also analyzed for urea by GLDH-method and creatinine by Jaffe-method using commercial kits to assesGlutathione contents were assessed by adapting the guidelines as mentioned by . The reaLiver function tests including aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were assessed. Levels of AST and ALT were measured by the dinitrophenylhydrazene (DNPH) method using Sigma Kits 59\u201350 and 58\u201350, respectively and ALP by Alkaline Phosphates\u2013DGKC method .Red blood cells indices including total red blood cells (TRBCs), hemoglobin (Hb), hematocrit (Hct) and mean corpuscular volume (MCV) were estimated. Likewise, white blood cell indices including monocytes, lymphocytes and neutrophils were measured by using Automatic Blood Analyzer .Statistical program SAS was utilized to analyze the data collected from this study. Two-way analysis of variance (ANOVA) was conducted to evaluate the effect of extraction time and solvent on polyphenol isolation. Moreover, One-way ANOVA were applied in efficacy trial and significance among the treatments were determined by applying LSD.p\u2009\u2265\u20090.001) and time (p\u2009\u2265\u20090.002), highest theaflavin (3.42\u2009\u00b1\u20090.27\u00a0g/100\u00a0g) were detected in ethanolic extract followed by methanol (2.77\u2009\u00b1\u20090.25\u00a0g/100\u00a0g) whilst water exhibited the lowest yield (2.32\u2009\u00b1\u20090.13\u00a0g/100\u00a0g). Similarly, solvent (p\u2009\u2265\u20090.001) & time (p\u2009\u2265\u20090.001) also imparted pronounced impact for thearubigins, highest in ethanol and minimum in water. Extraction efficiency was also influenced by time and maximum yield for theaflavins and thearubigins was obtained at 60\u00a0min 3.41\u2009\u00b1\u20090.17\u00a0g/100\u00a0g and 13.17\u2009\u00b1\u20090.59\u00a0g/100\u00a0g, respectively (Table\u00a0Means for extraction yield illuminated the significant effect of solvents p\u2009\u2265\u20090.00 and timep\u2009\u2265\u20090.001) and extraction time (p\u2009\u2265\u20090.0031) whilst their interactive effect showed non-significant trend (p\u2009\u2265\u20090.521). Means for TF1 indicated highest value (2.18\u2009\u00b1\u20090.06\u00a0mg/g) in ethanolic extract followed by methanol (1.96\u2009\u00b1\u20090.03\u00a0mg/g) and water (0.751\u2009\u00b1\u20090.001\u00a0mg/g). However, effect of time revealed maximum output (1.79\u2009\u00b1\u20090.006\u00a0mg/g) at 60\u00a0min and minimum (1.48\u2009\u00b1\u20090.001\u00a0mg/g) at 30\u00a0min. Likewise trend was observed for TF2A, TF2B & TF3, among the solvents highest ratio in ethanolic fractions and minimum in water and among time intervals maximum in 60\u00a0min and minimum in 30\u00a0min , \u00df-carotene (p\u2009\u2265\u20090.004) and FRAP (p\u2009\u2265\u20090.000) whilst non-significant differences (p\u2009\u2265\u20090.185) were observed for glucose diffusion. Means for DPPH in theaflavin and thearubigins indicated highest activity in ethanolic extract 83.27\u2009\u00b1\u20091.23 & 68.72\u2009\u00b1\u20093.61%, respectively followed by methanol and water. For the time factor, the highest DPPH activity of theaflavin was 82.69\u2009\u00b1\u20095.10% at 60\u00a0min whilst 78.03\u2009\u00b1\u20094.31 & 75.89\u2009\u00b1\u20093.92% at 90 and 30\u00a0min, respectively. Similarly, thearubigins also exhibited highest DPPH activity at 60\u00a0min followed by the methanolic (89.96\u2009\u00b1\u20092.31%) and water extract (87.02\u2009\u00b1\u20093.12%). Likewise, in thearubigins the values for this parameter in respective extracts were 87.71\u2009\u00b1\u20096.12, 84.92\u2009\u00b1\u20096.42 and 83.02\u2009\u00b1\u20097.12%. Considering time factor, highest glucose diffusion was reported at 60\u00a0min for the both parameters (Table\u00a0p\u2009\u2265\u20090.001) of experimental diets on weight in both control and renal malfunctional rats and recorded values for weight in renal malfunctional rats at the termination of study were 213.10\u2009\u00b1\u20096.11 & 213.73\u2009\u00b1\u20098.12, 217.74\u2009\u00b1\u20095.61 & 220.52\u2009\u00b1\u20099.26 and 214.17\u2009\u00b1\u20094.25 & 215.73\u2009\u00b1\u20096.23\u00a0g/rat for T1 T2 and T3, respectively effect on serum cholesterol level of rats, highest cholesterol level was observed in T0 (86.31\u2009\u00b1\u20094.63\u00a0mg/dL) that significantly reduced in T1 (81.14\u2009\u00b1\u20092.12\u00a0mg/dL) trailed by T3 (82.29\u2009\u00b1\u20094.25\u00a0mg/dL) and T2 (82.99\u2009\u00b1\u20095.63\u00a0mg/dL). In contrary, the tested diets imparted non-significant (p\u2009\u2265\u20090.091) enhancement in HDL levels of study animals. In this context, T0 drink consuming group exhibited least HDL value as 38.54\u2009\u00b1\u20092.36\u00a0mg/dL, respectively that elevated non-significantly to 39.70\u2009\u00b1\u20092.32, 38.92\u2009\u00b1\u20091.09 and 39.64\u2009\u00b1\u20091.56\u00a0mg/dL in T1, T2 and T3 groups, respectively. As for as LDL levels were concern, the tested diets caused momentous (p\u2009\u2265\u20090.000) decline on this trait. The maximum LDL was noticed in T0 that reduced substantially to in T1, T2 and T3, correspondingly. Triglycerides values for T0, T1, T2 and T3 differed non-momentously (p\u2009\u2265\u20090.128) i.e. 67.15\u2009\u00b1\u20094.20, 64.96\u2009\u00b1\u20095.15, 65.20\u2009\u00b1\u20094.12 and 65.13\u2009\u00b1\u20092.12\u00a0mg/dL (Table The prepared diets caused significant ( 86.31\u2009\u00b1\u2009.63\u00a0mg/dLp\u2009\u2265\u20090.000) impact of experimental diets on blood glucose levels of tested groups. In renal malfunctional rats, the highest glucose level 89.18\u2009\u00b1\u20096.14 in T0 group however, black tea polyphenols supplemented diets lowered the glucose value to 84.70\u2009\u00b1\u20096.13, 85.60\u2009\u00b1\u20093.20 and 83.98\u2009\u00b1\u20097.14\u00a0mg/dL in T1, T2 and T3 groups, correspondingly. Likewise, insulin also varied significantly (p\u2009\u2265\u20090.004) minimum insulin values 7.76\u2009\u00b1\u20090.22 \u03bcU/mL were observed in T0 whilst highest level 8.11\u2009\u00b1\u20090.12 \u03bcU/mL in T3 followed by 8.06\u2009\u00b1\u20090.41 \u03bcU/mL in T1 and 7.96\u2009\u00b1\u20090.52 \u03bcU/mL in T2 impact on these traits however, the diet supplemented with both theaflavin and thearubigins caused maximum effect in comparison with others. The highest urea level 37.83\u2009\u00b1\u20092.46 was recorded in T0 that suppressed to 33.44\u2009\u00b1\u20091.45, 34.09\u2009\u00b1\u20092.56 and 34.95\u2009\u00b1\u20092.91\u00a0mg/dL in T3, T1 and T2 groups. Likewise, highest creatinine level 1.15\u2009\u00b1\u20090.03\u00a0mg/dL was recorded in T0 group (control drink) that significantly suppressed to 1.04\u2009\u00b1\u20090.05\u00a0mg/dL in T3 (drink containing theaflavin+thearubigins), 1.08\u2009\u00b1\u20090.01\u00a0mg/dL in T1 (drink containing theaflavin) and 1.09\u2009\u00b1\u20090.02\u00a0mg/dL in T2 (drink containing thearubigins).The means in Table 0 group showed decreased glutathione content 43.12\u2009\u00b1\u20093.01\u00a0mg/L that momentously (p\u2009\u2265\u20090.001) enhanced to 46.19\u2009\u00b1\u20092.56, 44.91\u2009\u00b1\u20094.14 and 47.87\u2009\u00b1\u20094.12\u00a0mg/L in T1, T2 and T3 groups. Similar significant (p\u2009\u2265\u20090.000) effect of experimental diets was observed for MDA, the recorded values for this trait in T0 was 8.01\u2009\u00b1\u20090.21that differed significantly in T1, T2 and T3 by 6.78\u2009\u00b1\u20090.24, 7.55\u2009\u00b1\u20090.01 and 6.95\u2009\u00b1\u20090.4\u00a0mmol/L (Table T0 (49.06\u2009\u00b1\u20093.01\u00a0IU/L) was significantly ((p\u2009\u2265\u20090.001)) varied in T1 T2 and T3 groups with mean values 48.50\u2009\u00b1\u20092.42, 49.12\u2009\u00b1\u20092.13 and 46.38\u2009\u00b1\u20092.45\u00a0IU/L. Moreover, T0 group showed maximum AST level 105.27\u2009\u00b1\u20095.14\u00a0IU/Lthat reduced substantially ((p\u2009\u2265\u20090.002) in T1 (101.68\u2009\u00b1\u20099.12\u00a0IU/L), T2 (103.00\u2009\u00b1\u20099.02\u00a0IU/L) and T3 (109.01\u2009\u00b1\u20099.45\u00a0IU/L). Likewise, ALP level in T0 (196.89\u2009\u00b1\u200917.23\u00a0IU/L) was significantly ((p\u2009\u2265\u20090.001)) higher than that of T1 (177.89\u2009\u00b1\u200915.20\u00a0IU/L), T2 (180.90\u2009\u00b1\u200913.25\u00a0IU/L) and T3 (167.68\u2009\u00b1\u20099.62\u00a0IU/L).Mean values in Table 0, T1, T2 and T3, respectively. Mean Hb level in T0, T1, T2 and T3 were 10.88\u2009\u00b1\u20090.71, 11.15\u2009\u00b1\u20090.61, 10.98\u2009\u00b1\u20090.81 and 11.20\u2009\u00b1\u20090.71\u00a0g/L, respectively. Moreover, the recorded hematocrit value for T0 (36.79\u2009\u00b1\u20091.52%) was improved non-significantly in T1, T2 and T3 groups as 37.25\u2009\u00b1\u20092.71, 36.99\u2009\u00b1\u20092.12 and 37.03\u2009\u00b1\u20092.81%, respectively. Likewise, mean MCV values for T0, T1, T2 and T3 were 49.96\u2009\u00b1\u20092.48, 51.03\u2009\u00b1\u20093.82, 50.92\u2009\u00b1\u20093.97 and 51.65\u2009\u00b1\u20093.30\u00a0fl, respectively. Similarly, the mean WBCs in T0 were 17.29\u2009\u00b1\u20090.58 cells/nL that non-significantly decreased to 16.99\u2009\u00b1\u20090.49, 17.26\u2009\u00b1\u20090.65 and 16.85\u2009\u00b1\u20090.35 cells/nL, respectively in tested groups. Likewise, means for Neutrophils in T0, T1, T2, and T3 were 62.25\u2009\u00b1\u20091.28, 63.85\u2009\u00b1\u20092.82, 62.45\u2009\u00b1\u20093.25 and 64.63\u2009\u00b1\u20092.63%, respectively. Mean monocytes values for T0, T1, T2 and T3 groups were 5.29\u2009\u00b1\u20090.21, 5.35\u2009\u00b1\u20090.22, 5.41\u2009\u00b1\u20090.25 and 5.65\u2009\u00b1\u20090.63%, respectively. In study IV, values for Lymphocytes were 33.92\u2009\u00b1\u20091.21, 35.25\u2009\u00b1\u20091.82, 34.91\u2009\u00b1\u20091.25 and 35.29\u2009\u00b1\u20091.63% in T0, T1, T2 and T3 group, respectively determination to enhance the authenticity of the results. Likewise, human efficacy trial for insulin sensitivity/resistance by different test like HOMA-insulin resistance, QUIKI, and Matsuda should be conducted to unveil the mechanistic concerns."} +{"text": "In the crystal, the disaccharide and water mol\u00adecules form layers parallel to the 13H24O9\u00b7H2O, a structural model for part of bacterial O-anti\u00adgen polysaccharides from Shigella flexneri and Escherichia coli, crystallizes with four independent disaccharide mol\u00adecules and four water mol\u00adecules in the asymmetric unit. The conformation at the glycosidic linkage joining the two rhamnosyl residues is described by the torsion angles \u03c6H of 39, 30, 37 and 37\u00b0, and \u03c8H of \u221232, \u221235, \u221231 and \u221232\u00b0, which are the major conformation region known to be populated in an aqueous solution. The hexo\u00adpyran\u00adose rings have the 1C4 chair conformation. In the crystal, the disaccharide and water mol\u00adecules are associated through O\u2014H\u22efO hydrogen bonds, forming a layer parallel to the bc plane. The layers stack along the a axis via hydro\u00adphobic inter\u00adactions between the methyl groups.The title compound, C Shigella flexneri , which are defined, respectively, by H1A\u2014C1A\u2014O2B\u2014C2B, C1A\u2014O2B\u2014C2B\u2014H2B and H1B\u2014C1B\u2014O7B\u2014C7B. These torsion angles are (I)H =39\u00b0, \u03c8H = \u221232\u00b0 and \u03c6H(C7) = 49\u00b0, (II) \u03c6H = 30\u00b0, \u03c8H \u221235\u00b0 and \u03c6H(C7) = 52\u00b0, (III) \u03c6H = 36\u00b0, \u03c8H = \u221231\u00b0 and \u03c6H(C7) = 51\u00b0, and (IV) \u03c6H = 37\u00b0, \u03c8H = \u221232\u00b0 and \u03c6H(C7) = 51\u00b0, where (I)\u2013(IV) correspond to the four independent disaccharide mol\u00adecules 1\u20134, respectively, in Fig.\u00a02H, \u03c8H and \u03c6H(C7) angles are 35\u2005(4), \u221233\u2005(2) and 51\u2005(1)\u00b0, respectively. The \u03c6H torsion angle is governed by the exo-anomeric effect and should be approximately 40\u00b0 for an \u03b1-l-sugar, which is also the case in the title rhamnose-containing disaccharide mixture solution at ambient temperature.The title compound was synthesized according to the published procedures and treated as riding with Uiso(H) = 1.2\u20131.5Ueq. The O\u2014H bond and H\u22efH distances in the water mol\u00adecules were restrained to 0.85\u2005(1) and 1.34\u2005(1)\u2005\u00c5, respectively. The orientation of each water mol\u00adecule was adjusted and restrained with additional DFIX commands using parameters derived from a solid state DFT optimization of the crystal structure.Crystal data, data collection and structural refinement details are summarized in Table\u00a0210.1107/S2056989019006935/is5512sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019006935/is5512Isup2.hklStructure factors: contains datablock(s) I. DOI: 1915954CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dihedral angle between the indole ring system and the nitro-substituted benzene ring is 37.64\u2005(16)\u00b0. In the crystal, mol\u00adecules are linked by O\u2014-H\u22efO and N\u2014H\u22efO hydrogen bonds, forming chains along [010]. In addition, weak C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions further link the structure into a three-dimensional network.The title mol\u00adecule adopts an 17H12N2O3\u00b7H2O comprises two mol\u00adecules of (E)-3-(1H-indol-2-yl)-1-(4-nitro\u00adphen\u00adyl)prop-2-en-1-one and a water mol\u00adecule. The main mol\u00adecule adopts an s-cis configuration with respect to the C=O and C=C bonds. The dihedral angle between the indole ring system and the nitro-substituted benzene ring is 37.64\u2005(16)\u00b0. In the crystal, mol\u00adecules are linked by O\u2014-H\u22efO and N\u2014H\u22efO hydrogen bonds, forming chains along [010]. In addition, weak C\u2014H\u22efO, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions further link the structure into a three-dimensional network. The optimized structure was generated theoretically via a density functional theory (DFT) approach at the B3LYP/6\u2013311\u2005G++ basis level and the HOMO\u2013LUMO behaviour was elucidated to determine the energy gap. The obtained values of 2.70\u2005eV and 2.80\u2005eV (DFT) are desirable for optoelectronic applications. The inter\u00admolecular inter\u00adactions were qu\u00adanti\u00adfied and analysed using Hirshfeld surface analysis.The asymmetric unit of the title compound, 2C In addition, the chalcone bridge consists of two different double bonds, C=C and C=O, which contribute to the conjugation of charge transfer, leading to their excellent structural and spectroscopic properties group as an acceptor group because the decrease of the resonance effect leads to substantial changes in \u03c0-electron delocalization in the ring level, providing information about the geometry of the mol\u00adecule. The optimized structure is shown in Fig.\u00a01b. The geometrical parameters are mostly within normal ranges, the slight deviations from the experimental values are due to the fact that the optimization is performed in isolated conditions, whereas the crystal environment and hydrogen-bonding inter\u00adactions affect the results of the X-ray structure \u2005\u00c5] and C9=C10 [1.310\u2005(5)\u2005\u00c5] bonds. The compound is twisted about the C10\u2014C11 bond with C9\u2014C10\u2014C11\u2014O1 torsion angle of \u221221.9\u2005(6)\u00b0. The corres\u00adponding torsion angle obtained from the DFT study is 0.08\u00b0. In addition, the mol\u00adecule is twisted about the C11\u2014C12 bond with an O1\u2014C11\u2014C12\u2014C13 torsion angle of 167.7\u2005(4)\u00b0 . The differences between the experimental and calculated values show that the inter\u00admolecular hydrogen bond involving the water mol\u00adecule does not affect the planarity of the compound. A previous study adopts an c). The enone group (O1/C9\u2013C11) with maximum deviation of 0.082\u2005(3)\u2005\u00c5 at C11 forms dihedral angles of 21.5\u2005(2) and 16.3\u2005(2)\u00b0 with the indole ring system and the nitro-substituted benzene ring, respectively.The overall conformation of the mol\u00adecule can be described by the dihedral angle formed by the indole ring system (N1/C1\u2013C8) and the nitro-substituted benzene (C12\u2013C17) ring with a value of 37.64\u2005(16)\u00b0 Fig.\u00a01c. The evia O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds involving the solvent water mol\u00adecule. The water mol\u00adecule is connected to the carbonyl group and indole ring system by inter\u00admolecular O1W\u2014H1OW\u22efO1i and N1\u2014H1A\u22efO1W hydrogen bonds . Furthermore, C9\u2014H9A\u22efCg1 inter\u00adactions are observed along the a-axis direction, completing the three-dimensional structure. Two of the anti-parallel mol\u00adecules are linked by \u03c0\u2013\u03c0 stacking inter\u00adactions involving the centroids (Cg2 and Cg3) of the C1\u2013C6 and C12\u2013C17 rings with a centroid\u2013centroid distance Cg2\u22efCg3 of 3.534\u2005(3)\u2005\u00c5. These \u03c0\u2013\u03c0 inter\u00adactions further stabilize the crystal structure.In the crystal, four symmetry-related mol\u00adecules are connected to each other s Table\u00a01, formingon Fig.\u00a02. In addis Table\u00a01 link thene Fig.\u00a03a. Furthns Fig.\u00a03a involvCrystalExplorer3.1 pair and the full fingerprint is outlined in grey plots were generated with ds Fig.\u00a04. The finds Fig.\u00a04. The H\u22efHde in . With the shape-indexed mapping, the C\u2014H\u22ef\u03c0 inter\u00adactions can be observed as a bright-red spot identified with black arrows in Fig.\u00a06b. The blue spots near the ring represent the reciprocal C\u2014H\u22ef\u03c0 inter\u00adactions.The presence of the C\u2014H\u22ef\u03c0 inter\u00adactions can be seen in the pale-orange spot inside the circle of black arrows on the Hirshfeld surface mapped over in Fig.\u00a06a. With Eg) between the highest occupied mol\u00adecular orbital (HOMO) and lowest unoccupied mol\u00adecular orbital (LUMO) is a crucial factor in elucidating the mol\u00adecular electrical transport properties. In the present study, the HOMO and LUMO were computed at the DFT/B3LYP/6-311G++ theoretical level and the respective plots of the frontier mol\u00adecular orbital are illustrated in Fig.\u00a07et al., 2014et al., 2015et al., 2018Frontier mol\u00adecular orbital analysis is a vital tool in the development of mol\u00adecular electronic properties. The energy gap -3-(2-methyl\u00adphen\u00adyl)-1-(4-nitro\u00adphen\u00adyl)prop-2-en-1-one ethan\u00adone (0.5\u2005mmol) and indole-2-carboxaldehyde (0.5\u2005mmol) was dissolved in methanol (20\u2005mL). Sodium hydroxide (NaOH) solution was then added dropwise under vigorous stirring. The reaction mixture was stirred for 5\u20136\u2005h at room temperature. The final precipitate was filtered, washed with distilled water and recrystallized by slow evaporation from acetone solution to obtain orange plate-shaped crystals.The title compound was synthesized Uiso(H) = 1.2Ueq(C). The water O atom was refined with half-occupancy. The O- and N-bound H atoms were located from difference-Fourier maps and refined freely.Crystal data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018014329/lh5883sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018014329/lh5883Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018014329/lh5883Isup3.cmlSupporting information file. DOI: 1846181CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The mol\u00adecules are connected N,N-di\u00admethyl\u00adformamide (1/1/1), C14H10O4S2\u00b7C7H5ClO2\u00b7C3H7NO, contains a mol\u00adecule each of 2,2\u2032-di\u00adthiodi\u00adbenzoic acid (DTBA), 2-chloro\u00adbenzoic acid (2CBA) and di\u00admethyl\u00adformamide (DMF). The DTBA mol\u00adecule is twisted [the C\u2014S\u2014S\u2014C torsion angle is 88.37\u2005(17)\u00b0] and each carb\u00adoxy\u00adlic group is slightly twisted from the benzene ring to which it is connected [CO2/C6 dihedral angles = 7.6\u2005(3) and 12.5\u2005(3)\u00b0]. A small twist is evident in the mol\u00adecule of 2CBA [CO2/C6 dihedral angle = 4.4\u2005(4)\u00b0]. In the crystal, the three mol\u00adecules are connected by hydrogen bonds with the two carb\u00adoxy\u00adlic acid residues derived from DTBA and 2CBA forming a non-symmetric eight-membered {\u22efHOCO}2 synthon, and the second carb\u00adoxy\u00adlic acid of DTBA linked to the DMF mol\u00adecule via a seven-membered {\u22efHOCO\u22efHCO} heterosynthon. The three-mol\u00adecule aggregates are connected into a supra\u00admolecular chain along the a axis via DTBA-C\u2014H\u22efO(hydroxyl-2CBA), 2CBA-C\u2014H\u22efO(hydroxyl-DTBA) and DTBA-C\u2014H\u22efS(DTBA) inter\u00adactions. Supra\u00admolecular layers in the ab plane are formed as the chains are linked via DMF-C\u2014H\u22efS(DTBA) contacts, and these inter-digitate along the c-axis direction without specific points of contact between them. A Hirshfeld surface analysis points to additional but, weak contacts to stabilize the three-dimensional architecture: DTBA-C=O\u22efH(phenyl-DTBA), 2CBA-Cl\u22efH(phenyl-DTBA), as well as a \u03c0\u2013\u03c0 contact between the delocalized eight-membered {\u22efHOC=O}2 carb\u00adoxy\u00adlic dimer and the phenyl ring of 2CBA. The latter was confirmed by electrostatic potential (ESP) mapping.The asymmetric unit of the three-component title compound, 2,2\u2032-di\u00adthiodi\u00adbenzoic acid\u20132-chloro\u00adbenzoic acid\u2013 In this three-component crystal, one of the carb\u00adoxy\u00adlic acid groups of the DTBA mol\u00adecule forms hydrogen bonds to DMF rather than to 2CBA. Herein, the crystal and mol\u00adecular structures of the title co-crystal solvate are described along with an analysis of the calculated Hirshfeld surfaces and a computational chemistry study.Recent bibliographic reviews have highlighted the rich coord\u00adination chemistry based on ligands derived from 2-mercapto\u00adbenzoic acid (2-MBA) . The asymmetric unit comprises 2,2\u2032-di\u00adthiodi\u00adbenzoic acid (DTBA), 2-chloro\u00adbenzoic acid (2CBA) and a di\u00admethyl\u00adformamide (DMF) solvent mol\u00adecule in a stoichiometric 1:1:1 ratio, as illustrated in Fig.\u00a01The title compound, (I)et al., 2007As anti\u00adcipated, crystallography reveals that the original 2-mercapto\u00adbenzoic acid underwent oxidation to yield a mol\u00adecule of DTBA, with the benzoic acid moieties being bridged through a di\u00adsulfide bond [S1\u2014S2 = 2.053\u2005(1)\u2005\u00c5]. The presence of carb\u00adoxy\u00adlic acid groups is confirmed by the disparity in the bond lengths for C8\u2014O4, O3 [1.317\u2005(4) and 1.229\u2005(4)\u2005\u00c5] and C21\u2014O6, O5 [1.326\u2005(4) and 1.209\u2005(4)\u2005\u00c5]. Both carb\u00adoxy\u00adlic acid groups (O3\u2014C8\u2014O4 and O5\u2014C21\u2014O6) are slightly twisted from the benzene rings (C9/C14 and C15/C20) to which they are bonded with the corresponding dihedral angles being 7.6\u2005(3) and 12.5\u2005(3)\u00b0, respectively. The C14\u2014S1\u2014S2\u2014C15 torsion angle is 88.37\u2005(17)\u00b0, indicating an almost orthogonal disposition between the benzene rings. The carbonyl-O3 and O5 atoms are oriented towards the di\u00adsulfide-S1 and S2 atoms with S1\u22efO3 and S2\u22efO5 distances of 2.713\u2005(2) and 2.661\u2005(3)\u2005\u00c5, respectively, and are indicative of hypervalent S\u2190O inter\u00adactions and 1.320\u2005(4), respectively. The carb\u00adoxy\u00adlic acid group is almost co-planar with the phenyl ring (C2\u2013C7) as seen in the dihedral angle of 4.4\u2005(4)\u00b0 between their planes. Similarly, co-planarity is also noted between the chloride atom and benzene ring plane with the r.m.s deviation from the least-squares plane through the seven non-hydrogen atoms being 0.027\u2005\u00c5.PLATON .The geometric parameters characterizing the inter\u00adatomic contacts in the crystal of (I)2 homosynthon, as well as discrete DTBA-C11\u2014H11\u22efS1(DTBA) inter\u00adactions. These lead to a supra\u00admolecular chain along the crystallographic a direction, as indicated in Fig.\u00a02b). Inter\u00adactions between the chains leading to a layer in the ab plane occur through DMF-C24\u2014H24C\u22efS2(DTBA) contacts, Fig.\u00a02c). The layers inter-digitate along the c-axis direction with only weak contacts between them as detailed in the next section.The resultant three-mol\u00adecule aggregates are connected by DTBA-C10\u2014H10\u22efO2(hydroxyl-2CBA) and 2CBA-C3\u2014H3\u22efO4(hydroxyl-DTBA) inter\u00adactions to form a non-symmetric, ten-membered {\u22efOCCCH}Crystal Explorer 17 dnorm maps of the respective mol\u00adecules in the aggregates are shown in Fig.\u00a03dnorm map signifying close contacts which origin\u00adate from DTBA-O\u2014H\u22efO(carbonyl-2CBA), DTBA-O\u2014H\u22efO(carbonyl-DMF), DTBA-C=O\u22efH(hydroxyl-2CBA) and DTBA-C=O\u22efH(DMF). Other red spots are observed through the dnorm map, albeit with relatively weak intensity. The contacts are consistent with those identified above except for some additional inter\u00adactions such as DTBA-C=O\u22efH(phenyl-DTBA), 2CBA-Cl\u22efH(phenyl-DTBA) as well as a \u03c0\u2013\u03c0 contact between the delocalized eight-membered {\u22efHOC=O}2 carb\u00adoxy\u00adlic dimer and the phenyl ring of 2CBA, Fig.\u00a03b). To validate the non-conventional \u03c0\u2013\u03c0 contact, the inter\u00adacting mol\u00adecules were subjected to electrostatic potential (ESP) mapping using Spartan\u201916 level of theory. The ESP mapping shows that the dimeric ring ranges from electropositive to neutral within the centre of the ring while the phenyl ring of 2CBA is mainly neutral indicating that the inter\u00adaction is mainly diffusive in nature, Fig.\u00a03c) and (d). As for the 2CBA and DMF mol\u00adecules, the corresponding dnorm maps (not shown) are reflective of their inter\u00adactions with the DTBA mol\u00adecule.The di + de \u223c2.20\u2005\u00c5), O\u22efH/H\u22efO , C\u22efH/H\u22efC , S\u22efH/H\u22efS , Cl\u22efH/H\u22efCl and other contacts (14.0%). Almost all of these contacts are shorter than their corres\u00adponding sum van der Waals radii, with H\u22efH, O\u22efH, C\u22efH, S\u22efH and Cl\u22efH being \u223c2.4, \u223c2.72, \u223c2.9, \u223c3.0 and \u223c2.95\u2005\u00c5, respectively.The two-dimensional fingerprint plots were generated to qu\u00adantify the close contacts identified on the Hirshfeld surfaces. The overall fingerprint plot of (I)di + de contact distance of \u223c2.18 and \u223c2.24\u2005\u00c5 for DTBA and 2CBA, respectively. The O\u22efH/H\u22efO contacts are the second most dominant contact for the individual mol\u00adecules which lead to the distinctive spikes in the corresponding decomposed fingerprint plots with a contribution of 26.4% for DTBA and 22.2% for 2CBA. Further delin\u00adeation of the contact shows that DTBA possesses about 11.1% of -H\u22efO- and 15.3% -O\u22efH- compared to 2CBA with 10.9 and 11.2% of the equivalent contacts, both with approximately the same di + de contact distance of \u223c1.70\u2005\u00c5 for DTBA and \u223c1.62\u2005\u00c5 for 2CBA. Additional contacts for DTBA and 2CBA are respectively dominated by C\u22efH/H\u22efC , S\u22efH/H\u22efS and Cl\u22efH/H\u22efCl . As for the DMF solvent mol\u00adecule, this exhibits a relatively different claw-like profile with several disproportional spikes observed in the fingerprint plot mainly owing to the asymmetric inter\u00adaction environment for the O\u22efH/ H\u22efO contact, in which the contribution of -O\u22efH- contact to the Hirshfeld surfaces is about 14.6% (di + de \u223c1.60\u2005\u00c5), while the -H\u22efO- contact is about 11.2% (di + de \u223c2.22\u2005\u00c5) that can be summed up to yield an overall 25.8%. The contribution of other short contacts is noted in decreasing order: H\u22efH , C\u22efH/ H\u22efC and H\u22efS , respectively.The DTBA and 2CBA mol\u00adecules display similar fingerprint patterns having a claw-like profile in the respective full fingerprint plots, implying the existence of nearly identical inter\u00adactions between the mol\u00adecules which is expected considering the similarity of their mol\u00adecular structures. Detailed analysis of the decomposed fingerprint plots shows that H\u22efH is the most prevalent contact for the mol\u00adecules, with the percentage contribution to the overall contacts of 29.7 and 25.0% and minimum Crystal Explorer 17, Table\u00a02Eint) of \u221273.2\u2005kJ\u2005mol\u22121. This energy is about one and a half-fold greater than the second strongest inter\u00adaction that occurs between DTBA-DMF [DTBA-O\u2014H\u22efO(carbonyl-DMF)/ DTBA=O\u22efH\u2014C-(DMF)] with an Eint = \u221245.9\u2005kJ\u2005mol\u22121. The disparity in energy is likely due the replacement of one O\u2014H\u22efO hydrogen bond with a C\u2014H\u22efO inter\u00adaction in the latter inter\u00adaction.The energy calculations through 2 and the 2CBA-benzene ring gives an energy of \u221215.9\u2005kJ\u2005mol\u22121 which is considered weak in nature. This indicates the energy is mainly dominated by dispersive forces, Table\u00a02Eint for other inter\u00adactions present in the crystal were also calculated and the results are summarized as in Table\u00a02\u22121 which can be considered weak.On the other hand, the \u03c0\u2013\u03c0 inter\u00adaction between the hydrogen bond-mediated dimer of (DTBA)c-axis direction. A relatively weaker dispersion force co-exists along with the main energy framework due to \u03c0\u2013\u03c0 inter\u00adactions which help to sustain the overall mol\u00adecular packing of (I)The energy frameworks of (I)ba), in (II) the two DTBA mol\u00adecules (DTBA-IIa and DTBA-IIb) form hydrogen bonds with each other, to yield a non-symmetric homosynthon, and with the two remaining carb\u00adoxy\u00adlic acid groups being hydrogen bonded to two 3CBA mol\u00adecules to give rise to a four-mol\u00adecule aggregate.A structural analogue of (I)Mercury ca 12.4%) which is approximately 6% greater than 2CBA in (I)ca 6.3%).Both (I)et al., 2016et al., 20062 synthon in carb\u00adoxy\u00adlic acid structures, i.e. 33%, emphasizing that this particular synthon can be readily disrupted in the presence of competing synthons solution of the ground mixture. M.p. 437.3\u2013438.9\u2005K. IR (cm\u22121): 3076 \u03bd(C\u2014H), 1678 \u03bd(C=O), 1473 \u03bd(C=C), 1426 \u03b4(C\u2014H), 736 \u03b4(C\u2014Cl).All chemical precursors were of reagent grade and used as received without further purification. 2-Mercapto\u00adbenzoic acid was mixed with 2-chloro\u00adbenzoic acid and ground for 15 mins in the presence of a few drops of methanol. The procedure was repeated three times. Colourless blocks were obtained through the careful layering of toluene (1\u2005ml) on an Uiso(H) set to 1.2\u20131.5Ueq(C). The oxygen-bound H atoms were located from difference Fourier maps and refined without constraint. Owing to poor agreement, one reflection, i.e. (4 2 2), was omitted from the final cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901900375X/hb7808sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901900375X/hb7808Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901900375X/hb7808Isup3.cmlSupporting information file. DOI: 1903993CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the components are linked by Owater\u2014H\u22efN, N\u2014H\u22efOwater and N\u2014H\u22efN hydrogen bonds, forming chains along the [100] direction. The chains are linked by C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds, forming layers parallel to the ab plane. Finally, the layers are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional structure.The asymmetric unit of the title compound contains two independent organic mol\u00adecules which differ primarily in the dihedral angle between the aromatic rings, 20H20N4O\u00b70.5H2O, contains two independent organic mol\u00adecules (1 and 2) and a water mol\u00adecule of crystallization. The two mol\u00adecules differ primarily in the dihedral angles between the aromatic rings, which are 7.79\u2005(7) and 29.89\u2005(7)\u00b0 in mol\u00adecules 1 and 2, respectively. In each mol\u00adecule there is intra\u00admolecular C\u2014H\u22efO hydrogen bond forming an S(6) ring motif. In mol\u00adecule 1 there is an intra\u00admolecular N\u2014H\u22ef\u03c0(pyrazole) inter\u00adaction and an intra\u00admolecular C\u2014H\u22ef\u03c0(pyrazole) inter\u00adaction present. Mol\u00adecule 1 is linked to mol\u00adecule 2 by a C\u2014H\u22ef\u03c0(benzene ring) inter\u00adaction. An intra\u00admolecular N\u2014H\u22efN hydrogen bond and an intra\u00admolecular C\u2014H\u22efN hydrogen bond are also present in mol\u00adecule 2. In the crystal, the three components are linked by Owater\u2014H\u22efN, N\u2014H\u22efOwater and N\u2014H\u22efN hydrogen bonds, forming chains along the [100] direction. The chains are linked by C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds, forming layers parallel to the ab plane. Finally, the layers are linked by C\u2014H\u22ef\u03c0 inter\u00adactions, forming a three-dimensional structure.The asymmetric unit of the title compound, C We report herein on its crystal and mol\u00adecular structures along with the Hirshfeld surface analysis.Pyrazole derivatives are biologically active heterocyclic compounds ring motif and 1.3596\u2005(16)\u2005\u00c5, respectively.The pyrazole ring (N1/N2/C2\u2013C4) in mol\u00adecule 1 is inclined to the benzene rings (C7\u2013C12 and C14\u2013C19) by 70.83\u2005(8) and 76.79\u2005(8)\u00b0, respectively. The corresponding dihedral angles in mol\u00adecule 2, involving the N5/N6/C22\u2013C24 pyrazole ring and the C27\u2013C32 and C34\u2013C39 benzene rings, are 68.47\u2005(8) and 81.91\u2005(8)\u00b0, respectively. In both mol\u00adecules there is an intra\u00admolecular C\u2014H\u22efO hydrogen bond forming an if Fig.\u00a01. In the In mol\u00adecule 1, an intra\u00admolecular N\u2014H\u22ef\u03c0(pyrazole) inter\u00adaction and an intra\u00admolecular C\u2014H\u22ef\u03c0(pyrazole) inter\u00adaction are present Fig.\u00a01. Mol\u00adecuwater\u2014H\u22efN and N\u2014H\u22efOwater hydrogen bonds, and by N\u2014H\u22efN hydrogen bonds, forming chains propagating along the a-axis direction; see Fig.\u00a03ab plane \u00adphen\u00adyl]acetamides gave many hits. A search for the substructure [2-(benzyl\u00adidene\u00adamino)\u00adphen\u00adyl]acetamide gave 19 hits, some of which are metal complexes. The structures most similar to the title compound include: N-(2-{[(2-hy\u00addroxy\u00adphen\u00adyl)methyl\u00adidene]amino}\u00adphen\u00adyl)-2,2-di\u00admethyl\u00adpropanamide aniline -N-(o-nitro\u00adbenzil\u00adidene)aniline -N-(m-nitro\u00adbenzil\u00adidene)aniline \u00b0 for POSPET, 16.2\u2005(2)\u00b0 for RIHHOF, 41.81\u2005(14)\u00b0 for RIHHUL and 11.2\u2005(4)\u00b0 in RIHJAT. The dihedral angles between the aromatic rings in the title compound are 7.79\u2005(7) and 29.89\u2005(7)\u00b0 in mol\u00adecules 1 and 2, respectively.A search of the Cambridge Structural Database -1-(2-hy\u00addroxy\u00adphen\u00adyl)ethyl\u00adidene]amino}\u00adphen\u00adyl)-2-meth\u00adoxy\u00adacet\u00adamide amino]\u00adphen\u00adyl}acet\u00adamides gave an inter\u00adesting hit, namely that for CrystalExplorer17.5 to 1.583 (blue) \u00c5. Fig.\u00a06dnorm mapped on the Hirshfeld surface.The Hirshfeld surface analyse was carried out using a (H\u22efH) illustrates the two-dimensional fingerprint of the points associated with hydrogen atoms. It is characterized by an end point that points to the origin and corresponds to id = ed = 1.08\u2005\u00c5, which indicates the presence of the H\u22efH contacts in this study (54%). Fig.\u00a08b (C\u22efH/H\u22efC) shows the contacts between the carbon atoms inside the surface and the hydrogen atoms outside the surface of Hirshfeld and vice versa (24%). The O\u22efH/H\u22efO (11.5%) plot shows two symmetrical wings on the left and right sides . The N\u22efH/H\u22efN inter\u00adactions (6.5%) are visualized in Fig.\u00a08d.Fig.\u00a07es Fig.\u00a08c. The NN-2-amino\u00adphenyl-5-methyl\u00adpyrazol-3-ylacetamide with 4-methyl\u00adbenzaldehyde in acetone (50\u2005ml) for 3\u2005h. The solvent was evaporated under vacuum, and then water was added. The precipitate formed was filtered under vacuum and purified through silica gel column chromatography using hexa\u00adne/ethyl acetate , yielding colourless rod-like crystals of the title compound (yield 63%).The title compound was prepared by stirring Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018017747/xu5954sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018017747/xu5954Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018017747/xu5954Isup3.cmlSupporting information file. DOI: 1885214CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules form stacks along the [001] direction. The crystal packing is further stabilized by O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and C\u2014H\u22efCl and C\u2014H\u22ef\u03c0 contacts.The title compound, C 14H12ClNO3, is a Schiff base that exists in the keto\u2013enamine tautomeric form and adopts a Z configuration. In the crystal, the dihedral angle between the planes of the benzene rings is 5.34\u2005(15)\u00b0. The roughly planar geometry of the mol\u00adecule is stabilized by a strong intra\u00admolecular N\u2014H\u22efO hydrogen bond. In the crystal, pairs of centrosymmetrically related mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds, forming R22(10) rings. Besides this, the mol\u00adecules form stacks along the [001] direction with C\u2014H\u22ef\u03c0 and C\u2014H\u22efCl contacts between the stacks. The inter\u00admolecular inter\u00adactions in the crystal were analysed using Hirshfeld surfaces. The most significant contribution to the crystal packing is from H\u22efH contacts (30.8%).The title compound, C Such compounds can exist in two tautomeric forms, viz. keto\u2013enamine (N\u2014H\u22efO) and phenol\u2013imine (N\u22efH\u2014O) . The whole mol\u00adecule is almost planar, with a dihedral angle of 5.34\u2005(15)\u00b0 between the benzene ring planes. The meth\u00adoxy C14 atom deviates from the plane of the C1\u2013C6 benzene ring by 0.038\u2005(4) \u00c5. The torsion angles C1\u2014C6\u2014N1\u2014C7 and N1\u2014C7\u2014C8\u2014C9 are 5.8\u2005(5) and \u22120.6\u2005(5)\u00b0, respectively. The planar mol\u00adecular conformation is stabilized by the intra\u00admolecular N1\u2014H2\u22efO2 hydrogen bond -(phenyl\u00adimino)\u00admeth\u00adyl]-benzene-1,2-diol fragment revealed eight hits where this fragment adopts the keto\u2013enamine tautomeric form and 21 hits where it exists as the phenol\u2013imine tautomer. Distinctive bond lengths in the title structure are the same within standard uncertainties as the corresponding bond lengths in the structures of 2-hy\u00addroxy-6-[(2-meth\u00adoxy\u00adphen\u00adyl)amino\u00admethyl\u00adene]cyclo\u00adhexa-2,4-dienone imino\u00admeth\u00adyl]benzene-1,2-diol and 5-chloro-2-meth\u00adoxy\u00adaniline , both in 15\u2005mL of ethanol, with subsequent stirring for 5\u2005h under reflux. Single crystals were obtained by slow evaporation of an ethanol solution .Uiso(H) = 1.2Ueq(C) or Uiso(H) = 1.5Ueq(C) for methyl H atoms. The O- and N-bound H atoms were located in a difference map and freely refined.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019002123/yk2119sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019002123/yk2119Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019002123/yk2119Isup3.cmlSupporting information file. DOI: 1886956CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Dear Editor,+ ROR\u2010gamma\u2010T+ innate lymphoid cell (ILC3) development.\u2212/\u2212 mice,The BTB\u2010ZF proteins play essential roles in the development of the immune system.\u2212/\u2212 mouse. According to the surface maker expression by flow cytometry, D1 is a DN1\u2010like cell line, which is CD44+CD25\u2212IL\u20107R\u03b1+. We sorted double positive (DP), CD4 single positive (SP) and CD8SP thymocytes from wild\u2010type C57BL/6 and IL\u20107R\u03b1 mutant mice mentioned above \u2010dependent thymic cell line derived from a p53\u03b1+Figure\u00a0A. D1 cel+ cells to establish the stable cell line (Figure\u00a0hi thymocytes in ScanT mice was also higher than that of their wild\u2010type littermates, although the percentage of TCR\u03b2hi thymocytes in ScanT mice was severely reduced (Figure\u00a0Next we investigated the effect of Zbtb1 on IL\u20107R\u03b1 expression. We transduced the D1 cells with either retroviral vector Mir\u2010CTR or Zbtb1\u2010overexpressing retrovirus, and sorted the GFPe Figure\u00a0B. Consise Figure\u00a0C. Interee Figure\u00a0D. The ILd Figure\u00a0E.Altogether, our results suggest that Zbtb1 and IL\u20107R\u03b1 signalling can regulate each other during T\u2010cell development. IL\u20107R\u03b1 signalling negatively regulate Zbtb1, and vice versa. Detailed molecular mechanisms by which Zbtb1 regulate IL\u20107R\u03b1 expression in T cells are still under investigation.The authors confirm that there are no conflicts of interest.\u00a0Click here for additional data file."} +{"text": "Arising from M. A. Bakar et al. Nature Commun. 10.1038/s41467-017-00720-3 (2017)1 A number of X-ray studies have reported gold compounds with short AuI\u00b7\u00b7\u00b7H contacts, but solid spectroscopic evidence for AuI\u00b7\u00b7\u00b7H bonding has been missing.1 Notably, during the revision of this work, Bourissou et al.2 and Straka et al.3 have provided evidence of true intramolecular AuI\u00b7\u00b7\u00b7H hydrogen bonds in [Cl\u2013Au\u2013L]+ complexes, where L is a protonated N-heterocyclic carbene. The studied compounds feature intramolecular AuI\u00b7\u00b7\u00b7H+\u2013N bonds detected by means of NMR2 and infrared spectroscopies.3Hydrogen bonding to gold(I) and its effect on the structure and dynamics of molecules have been a matter of long debate.4 reported compound 1 NMR resonances at respective H(C) nuclei in 1 highly deshielded with respect to precursor 2 auride-like\u00b7\u00b7\u00b7hydrogen bonding interaction. In addition, computational analysis of NMR chemical shifts reveals that the deshielding effects at respective hydrogen nuclei are not directly related to Au\u00b7\u00b7\u00b7H\u2013C hydrogen bonding in 1. It is well known that interactions of hydrogen with transition metals compounds may influence the 1H NMR shifts in unexpected ways.5Here, we show that the Au1, which is one of the four Au\u00b7\u00b7\u00b7H\u2013C contacts in the molecule as proposed in ref. 4. The calculated minimum Au\u00b7\u00b7\u00b7H distances (2.61\u20132.62\u2009\u00c5) are in excellent agreement with the reported ones (2.61\u20132.65\u2009\u00c5). To afford computational analysis of NMR chemical shifts9 (\u03b4), the P(Ph)2 groups in 1 and 2 were replaced by P(CH3)2 groups in model systems 1\u2032 and 2\u2032 .8 Notably, the absence of the bulky P(Ph)2 ligands causes rotation of the central phenyl groups away from the Au6 cluster. The Au\u00b7\u00b7\u00b7H distances increase from 2.6\u2009\u00c5 to 2.77\u2009\u00c5 and the Au\u2013H2\u2013C2 angles bend from 167\u00b0 to 144\u00b0 2 ligands that potentially stabilize the whole cluster via dispersion interactions among themselves.10 To avoid these undesirable changes in calculations, we fixed the core of 1 and 2 in optimization of 1\u2032 and 2\u2032 and only methyl groups were optimized.In the following, we analyze Au2\u00b7\u00b7\u00b7H2\u2013C2 contact in ule Fig.\u00a0. Computa1\u2032 shows a low ED (0.016 e.bohr\u22123) with positive Laplacian (0.037 e.bohr\u22125) at the line critical point (LCP) of Au2\u00b7\u00b7\u00b7H2 interaction. These values are\u00a0less than a half of those for reported Au\u00b7\u00b7\u00b7H+\u2013N bonds.6 Small electron exchange between Au2 and H2 of 0.07 e (e\u2009=\u2009electron) is consistent with a dispersive interaction.12 The direction of the charge transfer in Au2\u00b7\u00b7\u00b7H2 interaction is from Au2 to H2 in 1\u2032 have negative charge, about \u22120.15\u2009e analysis of 13 analysis of 1\u2032 reveals a weak Au2\u00b7\u00b7\u00b7H2\u2013C2 interaction channel is found between H2\u2013C2 and Au3 6p orbitals . This further points to a minimal stabilization effect of Au\u00b7\u00b7\u00b7H\u2013C bonding in 1.Extended transition state-natural orbitals for chemical valence (ETS-NOCV)als Fig.\u00a0. Notablyals Fig.\u00a0 is also 1H chemical shifts between 1 and 2 (1\u2032 and 2\u2032) are in excellent agreement with the experimental ones, for H2 \u03941\u20132\u2009=\u20094.4 ppm, \u03941\u2032\u22122\u2032\u2009=\u20093.6 ppm, and \u0394exp1\u20132\u2009=\u20094.4 ppm arises from the diamagnetic part of the NMR chemical shift, \u0394\u03b4dia, which can be rationalized only by a depletion of electron density (ED) at the H2 nuclei. Molecular orbital (MO) analysis of \u0394\u03b4dia identifies that main part \u0394\u03b4dia (1.3 of 1.6 ppm) originates from four Au\u2013P \u03c0-back-bonding MOs at H2, H4, and H6 nuclei4 given in brackets in Fig.\u00a0The calculated differences in 1\u2032\u22122\u2032 deshielding difference at H2 nuclei (2 ppm) is dominated by local Ramsey-type paramagnetic couplings15 between H2\u2013C2 \u03c3-bond with C2 2py* in 1\u2032, which is not possible in 2\u2032, increases the MO\u2009\u2194\u2009MO* overlap in orbital magnetic couplings.16 This leads to the ~0.5 ppm larger paramagnetic deshielding at H2 in 1\u2032 in this particular coupling in 2\u2032 contributes only by 0.2 ppm. An analogous mechanism is likely to be responsible also for the deshielding resonance at C2. The paramagnetic part of the \u0394ing Fig.\u00a0. Overall1 are an example of a weak auride-like\u00b7\u00b7\u00b7hydrogen interaction, with small overall stabilizing effect on the cluster structure. Instead, the stabilizing effect can be attributed to the dispersion interactions among the P(Ph)2 groups, as documented previously.10 Distinct \u03b4(1H) NMR deshielding of C\u2013H groups in contact with Au6 cluster in 1 as compared with the precursor 2 is due to (a) the differential ED at the H2 atom in 1 as compared to precursor 2, and (b) side-on orbital interactions between nearby Au3 atom and H\u2013C MOs that increase the efficiency of the local Ramsey-type deshielding paramagnetic couplings in molecule 1 as compared with corresponding couplings in precursor 2.We conclude that the short Au\u00b7\u00b7\u00b7H contacts in Supplementary Information"} +{"text": "It crystallizes in the non-centrosymmetric space group Pna21, with one mol\u00adecule in the asymmetric unit, and is constructed from a pair of aromatic rings [2-(tri\u00adfluoro\u00admeth\u00adyl)phenyl and tetra\u00adzole], which are twisted by 76.8\u2005(1)\u00b0 relative to each other because of significant steric hindrance of the tri\u00adfluoro\u00admethyl group at the ortho position of the benzene ring. In the crystal, very weak C\u2014H\u22efN and C\u2014H\u22efF hydrogen bonds and aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions link the mol\u00adecules into a three-dimensional network. To further analyse the inter\u00admolecular inter\u00adactions, a Hirshfeld surface analysis, as well as inter\u00adaction energy calculations, were performed.The title compound, C Their biological properties, including anti\u00adviral, anti\u00adcancer, anti-tuberculosis, anti\u00adfungal and anti\u00adoxidant activities have been shown by numerous studies \u03c0,\u03c3-complexes possessing non-linear optical properties 2(CF3SO3)2] 2]BF4 and [Cu(C11H9F3N4S)(NH2SO3)(MeOH)] based on 5-[(prop-2-en-1-yl)sulfan\u00adyl]-1-[2-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl]-1H-tetra\u00adzole (I)11H9F3N4S) have been reported recently 2]BF4 [dihedral angle = 78.0\u2005(1)\u00b0] and [Cu(C11H9F3N4S)(NH2SO3)(MeOH)] [85.5\u2005(1)\u00b0] as well as calculation of the inter\u00adaction energies were performed using CrystalExplorer . Fingerprint plots were produced to show the inter\u00admolecular surface bond distances with the regions highlighted for C\u2014H\u22efF and C\u2014H\u22efN inter\u00adactions. The contribution to the surface area for H\u22efH contacts is 19.8%.To further analyse the inter\u00admolecular inter\u00adactions between the mol\u00adecules of (I)\u22efF Fig.\u00a05b and C\u2014\u22efN Fig.\u00a05c inter\u00adCrystalExplorer. The total inter\u00admolecular energy is the sum of energies of four main components, viz. electrostatic, polarization, dispersion and exchange-repulsion factors of 1.057, 0.740, 0.871 and 0.618, respectively covers C\u2014H\u22efN and C\u2014H\u22efF inter\u00adactions with the neighbouring mol\u00adecule generated by the symmetry code \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, z\u00a0\u2212\u00a0\u22121 and are mainly dispersive in nature.The inter\u00adaction energies in (I)et al., 2016H-tetra\u00adzoles are known only as ligands in the structures of copper(I) and silver(I) \u03c0-complexes. In the crystal structures of bis\u00ad[\u03bc2-\u03b72-5--1-phenyl-1H-tetra\u00adzole]di\u00adaqua\u00addisilver bis(tetra\u00adfluoro\u00adborate) sulfan\u00adyl]-1H-tetra\u00adzole}di\u00adaquadicopper bis\u00ad(tetra\u00adfluoro\u00adborate) -5-[(prop-2-en-1-yl)sulfan\u00adyl]-1H-tetra\u00adzole}di\u00adaqua\u00addicopper bis\u00ad(tetra\u00adfluoro\u00adborate) ethanol solvate sulfan\u00adyl]-1-[2-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl]-1H-tetra\u00adzole}bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfonato)\u00addicopper {\u03b72-1--5-[(prop-2-en-1-yl)sulfan\u00adyl]-1H-tetra\u00adzole}copper(I)} sulfanyl group is slightly elongated to 1.35\u20131.38\u2005\u00c5, in comparison with noncoordinated olefin bond value. The other S-substituted 1-phenyl-1H-tetra\u00adzole-5-thiol structures in the Cambridge Structural Database have different alkyl substit\u00aduents, such as 2-naphthyl benzene was distilled in vacuo.The title compound was synthesized from 2-(tri\u00adfluoro\u00admeth\u00adyl)aniline by a multi-step reaction. Commercially available 2-(tri\u00adfluoro\u00admeth\u00adyl)aniline was dissolved in the minimum amount of benzene and treated with carbon di\u00adsulfide and tri\u00adethyl\u00adamine . The solution was cooled to 273\u2005K and left for 5\u2005d. After complete precipitation of the tri\u00adethyl\u00adammonium di\u00adthio\u00adcarbamate salt, the solution was filtered. The solid was washed with anhydrous ether and air-dried for about 10\u2005min. The salt was then dissolved in about 7.5\u2005ml of chloro\u00adform, treated with 1.4\u2005ml of tri\u00adethyl\u00adamine and cooled to 273\u2005K. To this solution was added ethyl chloro\u00adformate dropwise over a 15\u2005min period under intensive stirring. The resulting solution was stirred at 273\u2005K for 10\u2005min and allowed to warm to room temperature over 1\u2005h. The chloro\u00adform solution was washed with 3 3 and refluxed under intensive stirring until the suspension disappeared. The solution was cooled to room temperature and washed with TBME. The water fraction was separated and acidified with 3 M HCl (Caution! During the acidification beware of toxic HN3 gas). The sediment of 1-[2-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl]-1H-tetra\u00adzole-5-thiol was separated by filtration and used for alkyl\u00adation without further purification.The obtained iso\u00adthio\u00adcyanate was mixed with water (10\u2005ml) and NaNH-tetra\u00adzole-5-thiol was dissolved in a solution of KOH in ethanol (10\u2005ml). To the solution allyl bromide was added and the mixture was heated at 323\u2005K for 1\u2005h. The solvent was removed in vacuo and to the residue was added water (5\u2005ml) and di\u00adchloro\u00admethane (10\u2005ml). The di\u00adchloro\u00admethane was separated and removed to give the title compound. Colourless blocks of (I)1-[2-(Tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl]-11H , \u03b4, p.p.m. 8.03 , 7.98\u20137.88 , 7.71 , 5.94 , 5.36 , 5.18 , 3.98 . Analysis calculated for C11H9F3N4S: C, 46.15; H, 3.17; N, 19.57; S, 11.20; found: C, 45.97; H, 3.04; N, 19.49; S, 11.27.NMR Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019011459/hb7842sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019011459/hb7842Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019011459/hb7842Isup3.molSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019011459/hb7842Isup4.cmlSupporting information file. DOI: 1947200CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-imidazol-3-ium chloride (IOH\u00b7Cl) is a new imidazolium salt with a hy\u00addroxy functionality.The title compound, 1,3-bis\u00ad(4-hy\u00addroxy\u00adphen\u00adyl)-1 H-imidazol-3-ium chloride (IOH\u00b7Cl), C15H13N2O2+\u00b7Cl\u2212, is a new imidazolium salt with a hy\u00addroxy functionality. The synthesis of IOH\u00b7Cl was achieved in high yield via a two-step procedure involving a di\u00adaza\u00adbutadiene precursor followed by ring closure using tri\u00admethylchloro\u00adsilane and paraformaldehyde. The structure of IOH\u00b7Cl consists of a central planar imidazolium ring (r.m.s. deviation = 0.0015\u2005\u00c5), with out-of-plane phenolic side arms. The dihedral angles between the 4-hy\u00addroxy\u00adphenyl substituents and the imidazole ring are 55.27\u2005(7) and 48.85\u2005(11)\u00b0. In the crystal, O\u2014H\u22efCl hydrogen bonds connect the distal hy\u00addroxy groups and Cl\u2212 anions in adjacent asymmetric units, one related by inversion and one by the n-glide and ] counterparts, with inter\u00adplanar distances of 3.560\u2005(3) and 3.778\u2005(3)\u2005\u00c5. The only other noteworthy inter\u00admolecular inter\u00adaction is an O\u22efO (not hydrogen bonded) close contact of 2.999\u2005(3)\u2005\u00c5 between crystallographically different hy\u00addroxy O atoms on translationally adjacent mol\u00adecules .Imidazolium salts are common building blocks for functional materials and in the synthesis of N-heterocyclic carbene (NHC) as \u03c3-donor ligands for stable metal complexes. The title salt, 1,3-bis\u00ad(4-hy\u00addroxy\u00adphen\u00adyl)-1 NaOtBu and NaH) represent a versatile class of ligand systems for metal-center activation or stabilization in modern organic synthesis (Arduengo para-phenol substituents (C4\u2013C9/O1 and C10\u2013C16/O2) bonded to the imidazolium N atoms [N1\u2014C4 = 1.442\u2005(3)\u2005\u00c5 and N2\u2014C10 = 1.441\u2005(3)\u2005\u00c5]. The phenol groups are out-of-plane, forming dihedral angles with the imidazolium ring of 55.27\u2005(7) and 48.85\u2005(11)\u00b0 for rings C4\u2013C9 and C10\u2013C15, respectively. The hy\u00addroxy H-atom coordinates were refined freely and are slightly out-of-plane of their respective phenolic groups; the torsion angles are 9.1\u2005(19)\u00b0 for C6\u2014C7\u2014O1\u2014H1O and 11\u2005(2)\u00b0 for C12\u2014C13\u2014O2\u2014H2O.In the structure of IOH\u00b7Cl Fig.\u00a02, there a\u2212 anion at to two different IOH\u00b7Cl mol\u00adecules, one related by inversion and the other by the n-glide. These hydrogen bonds, viz. O1i\u2014H1Oi\u22efCl1 and O2ii\u2014H2Oii\u2014Cl1 and 21-screw [C3\u2014H3\u22efClv; symmetry code: (v) \u2212x\u00a0\u2212\u00a0y\u00a0+\u00a0z\u00a0+\u00a0b and Table\u00a01a\u2013f) qu\u00adantify the majority of inter\u00admolecular contacts as H\u22efH and C\u22efH . In these diagrams, the O\u2014H\u22efCl hydrogen bonds are indicated by sharp diagonal jutting spikes , while C\u2014H\u22efO inter\u00adactions give less-pronounced spikes . C\u22efC contacts, which are all as a result of \u03c0\u2013\u03c0 stacking, account for 6.6% of the inter\u00admolecular contacts .The most prominent inter\u00admolecular inter\u00adactions in the crystals of IOH\u00b7Cl are O\u2014H\u22efCl hydrogen bonds. These link the Cls Table\u00a01. Short ces Fig.\u00a07d, whilees Fig.\u00a07e. C\u22efC cts Fig.\u00a07f.et al., 2016) on the three-ring fragment of the title compound yielded over 600 hits, ranging from similar simple salts to metal complexes containing analogous NHC frameworks. A search with H atoms bonded to the three carbons of the imidazole ring gave 180 hits. Of these, 28 had mesityl substituents, including IHOQUS , and the unsubstituted phenyl analog IPh\u00b7ClO4 -1,4-diazabutadiene (1)Synthesis of the precursor : to a round-bottomed flask charged with 15\u2005ml of methanol, 4-phenolaniline was added and stirred until fully dissolved. Glyoxal was added to the reaction solution with stirring. Upon addition of glyoxal solution, 40 wt.% in H2O, a brown precipitate formed and the solution turned orange. The reaction was further stirred at room temperature for 5\u2005h and the solid was vacuum filtered and washed with cold methanol . Step 2, Synthesis of IOH\u00b7Cl: ethyl acetate (10\u2005ml) was pre-heated to 343\u2005K. To the hot solution was added (1) and paraformaldehyde . The reaction mixture was stirred until all of the paraformaldehyde had dissolved. To this was added a solution of tri\u00admethyl\u00adchloro\u00adsilane (TMSCl) in ethyl acetate (0.15\u2005ml) dropwise over 5\u2005min while stirring. The solution was stirred for 2\u2005h and then placed in a refrigerator (275\u2005K) overnight. The precipitate was collected by vacuum filtration and washed with cold ethyl acetate and ether until the filtrate was colorless, yielding a dark-orange solid . Crystals were grown by slow evaporation of a concentrated solution in acetone.The overall reaction for the synthesis of the title compound is depicted in Fig.\u00a01Uiso(H) = 1.5Ueq(O). Carbon-bound H atoms were included in calculated positions and refined using a standard riding model, with C\u2014H = 0.95\u2005\u00c5 and Uiso(H) = 1.2Ueq(C). Refinement progress was checked using an R-tensor .Crystal data, data collection, and structure refinement details are given in Table\u00a0210.1107/S2056989019011058/su5504sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019011058/su5504Isup2.hklStructure factors: contains datablock(s) I. DOI: 1946122CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title compound, the heterocyclic portion of the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit adopts a flattened-boat conformation, while the oxazolidine ring adopts an envelope conformation. The 2-carbon link to the oxazole ring is perpendicular to the best plane through the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit. In the crystal, the mol\u00adecules form stacks extending along the normal to (104) through \u03c0-stacking inter\u00adactions between the two carbonyl groups and inversion-related oxazole rings. Aromatic rings from neighbouring stacks inter\u00adcalate to form an overall layer structure. 20H16Cl2N2O3S, is built up from a di\u00adhydro\u00adbenzo\u00adthia\u00adzine moiety linked by \u2013CH\u2013 and \u2013C2H4\u2013 units to 2,4-di\u00adchloro\u00adphenyl and 2-oxo-1,3-oxazolidine substituents, where the oxazole ring and the heterocyclic portion of the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit adopt envelope and flattened-boat conformations, respectively. The 2-carbon link to the oxazole ring is nearly perpendicular to the mean plane of the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit. In the crystal, the mol\u00adecules form stacks extending along the normal to (104) with the aromatic rings from neighbouring stacks inter\u00adcalating to form an overall layer structure. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (28.4%), H\u22efCl/Cl\u22efH (19.3%), H\u22efO/O\u22efH (17.0%), H\u22efC/C\u22efH (14.5%) and C\u22efC (8.2%) inter\u00adactions. Weak hydrogen-bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Density functional theory (DFT) optimized structures at the B3LYP/ 6\u2013311\u2005G level are compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2014LUMO behaviour was elucidated to determine the energy gap.The title compound, C A puckering analysis of the heterocyclic portion of the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit gave the parameters QT = 0.1206\u2005(14)\u2005\u00c5, q2 = 0.1190\u2005(14)\u2005\u00c5, q3 = \u22120.0174\u2005(16)\u2005\u00c5, \u03c6 = 178.2\u2005(8)\u00b0 and \u03b8 = 98.4\u2005(8)\u00b0, indicating a flattened-boat conformation. A similar analysis for the oxazolidine ring C (O2/N2/C11\u2013C13) yielded q2 = 0.1125\u2005(18)\u2005\u00c5 and \u03c62 = 45.7\u2005(9)\u00b0, indicating an envelope conformation with atom C12 at the flap position and at a distance of 0.175\u2005(2)\u2005\u00c5 from the best plane of the other four atoms. The C9/C10 chain C is essentially perpendicular to the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit, as indicated by the C6\u2014N1\u2014C9\u2014C10 torsion angle of 90.61\u2005(19)\u00b0. In the heterocyclic ring B, the C1\u2014S1\u2014C8 [104.29\u2005(8)\u00b0], S1\u2014C8\u2014C7 [121.39\u2005(12)\u00b0], C8\u2014C7\u2014N1 [120.77\u2005(14)\u00b0], C7\u2014N1\u2014C6 [126.86\u2005(14)\u00b0], C6\u2014C1\u2014S1 [123.97\u2005(13)\u00b0] and N1\u2014C6\u2014C1 [121.60\u2005(15)\u00b0] bond angles are enlarged compared with the corresponding values in the closely related compounds (2Z)-2-(4-chloro\u00adbenzyl\u00adidene)-4-[2-(2-oxooxazoliden-3-yl)eth\u00adyl]-3,4-di\u00adhydro-2H-1,4-benzo\u00adthia\u00adzin-3-one, (II), -2-[(4-fluoro\u00adbenzyl\u00adidene]-4-(prop-2-yn-1-yl)-3,4-di\u00adhydro-2H-1,4-benzo\u00adthia\u00adzin-3-one, (III), -4-[2-eth\u00adyl]-2(phenyl\u00admethyl\u00adidene)-3,4-di\u00adhydro-2H-1,4-benzo\u00adthia\u00adzin-3-one, (IV), ly Fig.\u00a01. The benC ring at \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01 and between C13=O3 and the C ring at \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z through \u03c0-stacking inter\u00adactions between C7=O1 and the ] Figs. 2 and 3 \u25b8.CrystalExplorer17.5 analysis have a nearly symmetrical distribution of points, Fig.\u00a08c, with thin edges at de + di = 2.88\u2005\u00c5. The fingerprint plot delineated into H\u22efO/O\u22efH contacts (17.0%), Fig.\u00a08d, has a pair of characteristic wings with a pair of spikes with the tips at de + di = 2.48\u2005\u00c5. In the absence of C\u2014H\u22ef\u03c0 inter\u00adactions, the pair of wings in the fingerprint plot delineated into H\u22efC/C\u22efH contacts (14.5%) have a nearly symmetrical distribution of points, Fig.\u00a08e, with thick edges at de + di \u223c2.66\u2005\u00c5. The C\u22efC contacts (8.2%), Fig.\u00a08f, have an arrow-shaped distribution of points with the tip at de = di \u223c1.68\u2005\u00c5. Finally, the H\u22efS/S\u22efH and C\u22efCl/Cl\u22efC contacts , and are seen as pairs of wide and thin spikes with the tips at de + di = 3.30 and 3.60\u2005\u00c5, respectively.The overall two-dimensional fingerprint plot, Fig.\u00a08H Table\u00a01 contribu\u22efH Fig.\u00a08g and C\u22ef\u22efC Fig.\u00a08h contacdnorm plotted onto the surface are shown for the H\u22efH, H\u22efCl/Cl\u22efH, H\u22efO/O\u22efH, H\u22efC/C\u22efH, C\u22efC and H\u22efS/S\u22efH inter\u00adactions in Fig.\u00a09a\u2013f, respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing using standard B3LYP functional and 6\u2013311\u2005G basis-set calculations -2-[methyl\u00adidene]-4-[2-eth\u00adyl]3,4-di\u00adhydro-2H-1,4- benzo\u00adthia\u00adzin-3-one ring. The energy band gap [\u0394E = ELUMO\u00a0\u2212\u00a0EHOMO] of the mol\u00adecule is about 3.42\u2005eV, and the frontier mol\u00adecular orbital energies, EHOMO and ELUMO are \u22125.44 and \u22122.02\u2005eV, respectively.The optimized structure of the title compound, (I)et al., 2016II -one , 2.20 equiv. of bis\u00ad(2-chloro\u00adeth\u00adyl)amine hydro\u00adchloride and 2.00 equiv. of potassium carbonate were added to a solution of (Z)-2--2H-1,4-benzo\u00adthia\u00adzin-3(4H)-one (1.5\u2005mmol) in DMF (25\u2005ml). The mixture was stirred at 353\u2005K for 6\u2005h. After removal of salts by filtration, the solution was evaporated under reduced pressure and the residue obtained was dissolved in di\u00adchloro\u00admethane. The remaining salts were extracted with distilled water. The residue obtained was chromatographed on a silica gel column (eluent: ethyl acetate/hexa\u00adne: 3/2). The isolated solid was recrystallized from ethanol solution to afford colourless crystals [light yellow in CIF?] (yield: 67%).Tetra-Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019004250/lh5895sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019004250/lh5895Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019004250/lh5895Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019004250/lh5895Isup4.cmlSupporting information file. DOI: 1906476CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure determination of Na 10[Pd2Cl6](C4H9SO3)8\u00b74H2O, were obtained from a water/2-propanol solution of sodium n-butane\u00adsulfonate and sodium tetra\u00adchlorido\u00adpalladate(II). In the crystal, sodium n-butane\u00adsulfonate anions and water mol\u00adecules are arranged in an amphiphilic inverse bilayered cationic array represented by the formula {[Na10(C4H9SO3)8(H2O)4]2+}n. Within this lamellar array: (i) a hydro\u00adphilic layer region parallel to the bc plane is established by the Na+ cations, the H2O mol\u00adecules and the O3S\u2013 groups of the sulfonate ions, and (ii) hydro\u00adphobic regions are present containing all the n-butyl groups in an almost parallel orientation, with the chain direction approximately perpendicular to the aforementioned hydro\u00adphilic layer. Unexpectedly, the flat centrosymmetric [Pd2Cl6]2\u2212 anion in the structure is placed between the butyl groups, within the hydro\u00adphobic regions, but due to its appropriate length primarily bonded to the hydro\u00adphilic \u2018inorganic\u2019 layer regions above and below the hydro\u00adphobic area via Pd\u2014Clt\u22efNa- and Pd\u2014Clt\u22efH\u2014O(H)\u2014Na-type inter\u00adactions. In addition to these hydrogen-bonding inter\u00adactions, both aqua ligands are engaged in charge-supported S\u2014O\u22efH\u2014O hydrogen bonds of a motif characterized by the D43(9) graph-set descriptor within the hydro\u00adphilic region. The crystal structure of the title compound is the first reported for a metal n-butane\u00adsulfonate.In the course of crystal-engineering experiments, crystals of the hydrated title salt, Na In contrast to methane\u00adsulfonates (n = 1) and ethane\u00adsulfonates (n = 2), there is only rare structure information for the next higher homologues chemistry.Sodium alkane\u00adsulfonates are artificial soaps (anionic tensides) with a widespread use having the typical brown colour of palladium complexes with a square-planar coordination environment. According to the results of elemental analysis and vibrational spectroscopic investigations, hydrated sodium cations, n-butane\u00adsulfonate and hexa\u00adchlorido\u00addipalladate(II) anions are present in the solid. The crystal structure determination of this compound is the first of a metal n-butane\u00adsulfonate and eventually confirmed the composition Na10(C4H9SO3)8[Pd2Cl6]\u00b74H2O and a lamellar amphiphilic structure.In the investigation described herein, the incorporation of hexa\u00adchlorido\u00addipalladate(II) anions into the sodium n-butane\u00adsulfonate anions and, close to a center of inversion, one half of a hexa\u00adchlorido\u00addipalladate anion. The five Na+ cations are in quite different coordination environments . Bond lengths and angles of the n-butanesulfonate anions are as expected (see supplementary Tables). All these anions are found with an entirely anti-periplanar conformation of the alkyl groups, without any disorder. Altogether, n-butane\u00adsulfonate anions, Na+ cations and water mol\u00adecules form a tenside-like inverse bilayered cationic array, which can be described by the formula {[Na10(H2O)4(C4H9SO3)8]2+}n. In this arrangement, the layer-like regions are oriented parallel to the bc plane of the unit cell. As visualized by the blue and the red sections of the transparent background of Fig.\u00a03+ cations, the H2O mol\u00adecules serving as aqua ligands in \u03bc bridging mode coordination, and the O3S\u2013 groups of the sulfonate ions. With all the C4-chains in an approximately parallel orientation, the butyl groups are arranged on both sides of the hydro\u00adphilic region to complete the amphiphilic double layer with an inverse bilayer thickness according to unit-cell parameter a. The centrosymmetric [Pd2Cl6]2\u2212 anions in the structure of 1 are placed between the n-butyl groups within the hydro\u00adphobic regions. In a first view, this position seems to be unexpected; however, the length of the dipalladate(II) anion is appropriate to allow for pronounced bonding to the hydro\u00adphilic \u2018inorganic\u2019 layered regions above and below the hydro\u00adphobic area and 2.3212\u2005(12)\u2005\u00c5]. These geometric parameters, as well as the Cl\u2014Pd\u2014Cl bond angles of 86.20\u2005(4) to 92.45\u2005(4)\u00b0 and the Pd\u2014\u03bc-Cl\u2014Pd angle of 93.80\u2005(4)\u00b0, are in good agreement with those found in Cs2[Pd2Cl6] : Na8(C4H9SO3)8\u00b7Na2Pd2Cl6\u00b74H2O. This choice takes into account that the Na\u2014Cl distance from the terminal chlorido ligand Cl2 of the hexa\u00adchlorido\u00addipalladate(II) anion to the sodium cation Na1 [2.8560\u2005(18)\u2005\u00c5] is close to the distances of 2.809\u2005(3) to 2.821\u2005(2)\u2005\u00c5 in Na2PdCl4 and 2.2800\u2005(10)\u2005\u00c5] are slightly shorter than the Pd\u2014+ cations , the two crystallographically independent water mol\u00adecules O1 and O2 in 1 are engaged in non-covalent bonding within the hydro\u00adphilic region (Table\u00a012Cl6]2\u2212 anion [D\u22efA distance = 3.127\u2005(3)\u2005\u00c5] and a charge-supported weak O\u2014H\u22efO type hydrogen bond to an O atom of a sulfonate anion containing S4 [D\u22efA = 2.879\u2005(4)\u2005\u00c5]. In contrast, the water mol\u00adecule containing O2 is engaged in two O\u2014H\u22efO type hydrogen bonds to sulfonate ions, one of moderate strength to an O atom of the sulfonate ion containing S4 [D\u22efA = 2.723\u2005(4)\u2005\u00c5] and a weak one to an O atom of the sulfonate ion containing S3 [D\u22efA = 2.884\u2005(4)\u2005\u00c5]. Pd\u2014Clterm\u22efH\u2014O(Na2)\u2014H\u22efO14(S4)\u22efH\u2014O(Na2)\u2014H\u22efO\u2014S is the entire path of hydrogen bonding described by the D34(9) graph-set descriptor with n = 1\u20134 gave three hits, viz. the structures of sodium methane\u00adsulfonate anion results in 46 entries. However, from a structural point of view, the role of the [Pd2Cl6]2\u2212 ion in 1 is completely different from the role of this species in all the other compounds. In addition to the reports on these compounds having organic components, there is one report on an inorganic ternary chloride containing the [Pd2Cl6]2\u2212 ion of sodium n-butane\u00adsulfonate and 1.177\u2005g (4\u2005mmol) of sodium tetra\u00adchlorido\u00adpalladate(II). The evaporation temperature of the solution was adjusted to 288\u2005K with a thermostat. After three days, crystals suitable for X-ray crystal structure determination could be harvested . A single crystal was selected directly from the mother liquor. Raman spectroscopy was done with a Bruker MultiRAM spectrometer, equipped with a Nd:YAG laser (1064\u2005nm) and an InGaAs detector (4000\u201370\u2005cm\u22121): \u03bd(C\u2014H): 2969 (m), 2920 (s), 2872 (m); \u03b4s(C\u2014H): 1445 (w), 1412 (w); \u03b4as(C\u2014H): 1306 (w); \u03bdas(S\u2014O): 1071 (s); \u03bds(C\u2014S): 800 (m); \u03b4(S\u2014O): 551 (m), 536 (m); \u03bd(Pd\u2014Clterm): 343 (m), \u03bd(Pd\u2014\u03bc-Cl): 305 (s); \u03bd(Pd\u2014\u03bc-Cl): 273 (m). Band assignments were made according to Fujimori and an universal ATR equipment: \u03bd(O\u2014H): 3503 (s), 3462 (sh), 3436 (s), 3367 (s); \u03bd(C\u2014H): 2967 (s), 2936 (s), 2872 (m); \u03b4(O\u2014H): 1662 (m), 1602 (m); \u03b4s(C\u2014H) 1465 (m), 1412 (w), 1378 (w), \u03b4as(C\u2014H): 1314 (w), 1286 (w); \u03bdas(C\u2014H): 1241 (w); \u03bds(S\u2014O): 1190 (s), 1166 (s); \u03bdas(S\u2014O): 1057 (s), 1044 (s); \u03bds(C\u2014S): 794 (m); \u03b4(S\u2014O): 555 (m), 534 (m); band assignment according to Fujimori (195932H80Cl6Na10O28Pd2S8 (1824.84\u2005g\u2005mol\u22121): C 21.06, H 4.42, S 14.06; found: C 20.78, H 4.49, S 12.98.Thin brown platelets of mori 1959 and Geri al. 1997. An IR smori 1959. A CHS a3 groups were allowed to rotate around the neighboring C\u2014C bonds. The Uiso(H) values were set to 1.5Ueq(Cmeth\u00adyl) and 1.2Ueq(Cmethyl\u00adene), respectively. H\u2014O distances of the water mol\u00adecules were restrained to 0.83\u2005(3)\u2005\u00c5.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019004201/wm5491sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019004201/wm5491Isup2.hklStructure factors: contains datablock(s) I. DOI: 1906335CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Furthermore, ADAM10 participates to spine shaping through the cleavage of adhesion molecules and its activity is under the control of synaptic plasticity events. In particular, long-term depression (LTD) promotes ADAM10 synaptic localization triggering its forward trafficking to the synapse, while long-term potentiation elicits ADAM10 internalization. Here, we show that a short-term in vitro exposure to A\u03b21\u201342 oligomers, at a concentration capable of inducing synaptic depression and spine loss, triggers an increase in ADAM10 synaptic localization in hippocampal neuronal cultures. However, the A\u03b21\u201342 oligomers-induced synaptic depression does not foster ADAM10 delivery to the synapse, as the physiological LTD, but impairs ADAM10 endocytosis. Moreover, A\u03b21\u201342 oligomers-induced inhibition of ADAM10 internalization requires neuronal activity and the activation of the NMDA receptors. These data suggest that, at the synaptic level, A\u03b21\u201342 oligomers trigger an aberrant plasticity mechanism according to which A\u03b21\u201342 oligomers can downregulate A\u03b2 generation through the modulation of ADAM10 synaptic availability. Moreover, the increased activity of ADAM10 towards its synaptic substrates could also affect the structural plasticity phenomena. Overall, these data shed new lights on the strict and complex relationship existing between synaptic activity and the primary mechanisms of AD pathogenesis.A disintegrin and metalloproteinase 10 (ADAM10) is a synaptic enzyme that has been previously shown to limit amyloid-\u03b2The online version of this article (10.1007/s12035-019-1583-5) contains supplementary material, which is available to authorized users. Then, to induce chemical LTD, we applied NMDA (20\u00a0\u03bcM) and glycine for 3\u00a0min at room temperature in MgQuantification of Western Blot analysis was performed by means of computer-assisted imaging . The levels of the proteins were expressed as relative optical density (OD) measurements and normalized on tubulin. Values are expressed as mean \u00b1 S.E.M. of at least three independent experiments.t test or, when appropriate, by using one-way ANOVA followed by Bonferroni\u2019s post hoc test or Kruskal-Wallis analysis of variance followed by Dunn\u2019s post hoc test.Co-localization analysis was performed using Zeiss AIM 4.2 software and spines analysis was performed with ImageJ software . For co-localization, and morphological analysis, cells were chosen randomly for quantification from 4 different coverslips (2\u20133 independent experiments), images were acquired using the same settings/exposure times, and at least 10 cells for each condition were analyzed. Statistical evaluations were performed by using 2-tailed Student\u2019s Suppl. Fig. 11\u201342 short exposure does not affect the expression and synaptic levels of GluN1, GluN2A, GluN2B and PSD-95. A) Quantitative analysis of Western Blot analysis of homogenate reported in Fig.42\u20131 70.59\u2009\u00b1\u200917.76%, oA\u03b21\u201342 109.3\u2009\u00b1\u200914.55%; GluN2A, CTRL 100\u2009\u00b1\u200924.25, A\u03b2 42\u20131 119.9\u2009\u00b1\u200934.25%, oA\u03b21\u201342 80.28\u2009\u00b1\u20098.62%; GluN2B, CTRL 100\u2009\u00b1\u200930.65%, A\u03b2 42\u20131 90.94\u2009\u00b1\u200916.17%, oA\u03b21\u201342 99.64\u2009\u00b1\u200945.05%; PSD-95, CTRL 100\u2009\u00b1\u200920.64%, A\u03b2 42\u20131 132.2\u2009\u00b1\u20096.41%, oA\u03b21\u201342 111\u2009\u00b1\u200928.41%; p\u2009>\u20090.05, one-way ANOVA, n\u2009=\u20095); B) Quantitative analysis of Western Blot analysis of TIF reported in Fig.42\u20131\u2009=\u200983.29\u2009\u00b1\u200932.03%, oA\u03b21\u201342 72.32\u2009\u00b1\u200913.9%; GluN2A CTRL 100\u2009\u00b1\u200916.60%, A\u03b242\u20131 106.9\u2009\u00b1\u200910.78%, oA\u03b21\u201342 104.9\u2009\u00b1\u200918.36%; GluN2B, CTRL 100\u2009\u00b1\u200925.43%, A\u03b242\u20131 117.3\u2009\u00b1\u200919.03, oA\u03b21\u201342 102.0\u2009\u00b1\u200914.97%; PSD-95, CTRL 100\u2009\u00b1\u20098.03, A\u03b242\u20131 106.2\u2009\u00b1\u20095.13, oA\u03b21\u201342 102\u2009\u00b1\u20095.84; p\u2009>\u20090.05, one-way ANOVA, n\u2009=\u20095). (PNG 43\u00a0kb)oA\u03b2High Resolution Image (TIF 130\u00a0kb)Suppl. Fig. 21\u201342 short exposure does not affect the synaptic localization of SAP97 and \u03b22-adaptin. A) Quantitative analysis of Western Blot analysis of SAP97 levels in TIF reported in Fig. 42\u20131 123.6\u2009\u00b1\u200929.07%, oA\u03b21\u201342 96.75\u2009\u00b1\u200921.64%; p\u2009>\u20090.05, one-way ANOVA, n\u2009=\u20095); B) Quantitative analysis of Western Blot analysis of \u03b22-adaptin in TIF reported in Fig.42\u20131\u2009=\u2009101.50\u2009\u00b1\u20099.20%, oA\u03b21\u201342 110.60\u2009\u00b1\u20098.35%; p\u2009>\u20090.05, one-way ANOVA, n\u2009=\u20095) (PNG 44\u00a0kb)oA\u03b2High Resolution Image (TIF 109\u00a0kb)Suppl. Fig. 31\u201342-induced increase in ADAM10 synaptic availability does not involve the activation of GluN2B-containing NMDA receptors. The presence of ifenprodil , an inhibitor of GluN2B-containing NMDA receptors, does not affect oA\u03b21\u201342-triggered augment in ADAM10 synaptic levels (PNG 66\u00a0kb)oA\u03b2High Resolution Image (TIF 134\u00a0kb)"} +{"text": "In comparison with previous determinations, the present redetermination results in improved precision of the structural parameters.Sr 2PdO3 was carried out using high-quality single-crystal X-ray data. The Sr2PdO3 structure has been described previously in at least three reports [Wasel-Nielen & Hoppe as reported previously. The structure consists of infinite chains of corner-sharing PdO4 plaquettes inter\u00adspersed by SrII atoms. A brief comparison of Sr2PdO3 with the related K2NiF4 structure type is given.The crystal structure redetermination of Sroppe 1970. Z. Anor Roy 1971. Adv. Ch al. 2002. J. Allo The PdII atom forms distorted PdO4 square planes, which are linked by sharing oxygen atoms in the trans-position to form infinite chains extending along the b-axis direction as shown in Fig.\u00a01j Wyckoff site having mm2 site symmetry. It is seven-coordinate in a monocapped trigonal\u2013prismatic fashion by oxygen with three different bond lengths \u00d7 3.7887\u2005(2) \u00d7 3.9822\u2005(3)\u2005\u00c53 \u2005\u00c5] and five SrII atoms with one short [2.474\u2005(2)\u2005\u00c5] and four long distances [2.6668\u2005(2)\u2005\u00c5] \u2005\u00c5] and two PdII atoms [1.9911\u2005(2)\u2005\u00c5] I. DOI: 10.1107/S2056989018017176/wm5474Isup2.hklStructure factors: contains datablock(s) I. DOI: 1882781CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Kip1 and p21Cip1, effectively canceled the accelerated activation of NF\u2010\u03baB, suggesting that oncogenic Ras\u2010induced cell cycle progression is essential for the hyperactivation of NF\u2010\u03baB. Furthermore, we found that Ras (G12V) augmented the transcriptional activation of NF\u2010\u03baB, and this activation required the p38 MAP kinase. We observed that a downstream kinase of p38 MAP kinase, MSK1, was activated by Ras (G12V) and catalyzed the phosphorylation of p65/RelA at Ser\u2010276, which is critical for its transcriptional activation. Significantly, phosphorylation of the p65/RelA subunit at Ser\u2010276 was elevated in patient samples of colorectal cancer harboring oncogenic mutations of the K\u2010Ras gene, and the expression levels of NF\u2010\u03baB target genes were drastically enhanced in several cancer tissues. These observations strongly suggest that oncogenic signal\u2010induced acceleration of NF\u2010\u03baB activation is caused by activation of the p38 MAP kinase\u2013MSK1 signaling axis and by cell cycle progression in cancer cells.It is well established that nuclear factor \u03baB (NF\u2010\u03baB) acts as one of the most important transcription factors for tumor initiation and progression, as it both protects cells from apoptotic/necrotic signals and accelerates angiogenesis and tumor metastasis, which is mediated via the expression of target genes. However, it has not yet been clarified how oncogenic signals accelerate the activation of NF\u2010\u03baB. In the current study, we utilized untransformed NIH\u20103T3 cells stably harboring a \u03baB\u2010driven luciferase gene to show that an oncogenic mutant of Ras GTPase augmented TNF\u03b1\u2010induced NF\u2010\u03baB activation. Notably, enforced expression of cyclin\u2010dependent kinase inhibitors, such as p27 CDKcyclin\u2010dependent kinaseCHXcycloheximideCOX\u20102cyclooxygenase 2CRCcolorectal cancerDMEMDulbecco's modified Eagle's mediumGAPGTPase\u2010activating proteinICAM\u20101intercellular adhesion molecule\u20101I\u03baB\u03b1inhibitor of \u03baB\u03b1MSK1/2mitogen and stress activated protein kinase 1/2NF\u2010\u03baBnuclear factor \u03baBNPMnucleophosminPI\u20103 kinasephosphatidylinositol\u20103 kinasePKAprotein kinase ARHDRel homology domainshRNAshort\u2010hairpin RNATNF\u03b1tumor necrosis factor \u03b1VEGFvascular endothelial growth factor1et al., Nuclear factor \u03baB (NF\u2010\u03baB) includes a family of transcription factors, which induce the expression of a number of target genes related to anti\u2010inflammation, development, and the apoptosis\u2010related responses supplemented with 10% FBS, 2\u00a0m2.2Kip1, and p21Cip1 were amplified by PCR using PrimeSTAR GXL DNA polymerase and inserted into EcoRI and XhoI sites of MSCV\u2010ires\u2010Puro retroviral vector.pBabePuro\u2010H\u2010Ras (G12V) was a kind gift from S. Lowe, Sloan Kettering Institute. pBabePuro\u2010B\u2010RAF (V600E) was a gift from W. Hahn (Addgene plasmid # 15269). cDNA of p16Ink4a, p272.3An anti\u2010FLAG antibody (M2) was purchased from Merck Millipore . Anti\u2010cyclin A (SC\u2010751), anti\u2010cyclin D1 (SC\u2010753), anti\u2010CDC2 (SC\u2010163), anti\u2010CDK4 (SC\u2010601), anti\u2010H\u2010Ras (SC\u2010520), and anti\u2010\u03b2\u2010actin (SC\u2010130301) antibodies were obtained from Santa Cruz Biotechnology . An anti\u2010phospho p65/RelA (S276) antibody was purchased from Rockland Immunochemicals . Anti\u2010MSK1 and anti\u2010MSK2 antibodies were purchased from Cell Signaling Technology and BD Biosciences , respectively. TNF\u03b1 and PDGF were purchased from PeproTech . LY294002, U0126, SP600125, and SB203580 were obtained from FUJIFILM Wako Pure Chemical . NOX inhibitor, VAS2870, was purchased from Merck Millipore .2.46 cells per 60\u2010mm\u2010diameter culture dish) with helpers such as pE\u2010Eco and pGP (TAKARA\u2010Bio). Twenty\u2010four hours later, culture medium was replaced with 1.5\u00a0mL fresh culture medium. Secreted retroviruses were harvested every 4\u20136\u00a0h during 24\u201360\u00a0h post\u2010transfection, pooled, and stored on ice. Exponentially growing cells (1\u00a0\u00d7\u00a0105 cells per 60\u2010mm\u2010diameter culture dish) were infected several times at 2\u2010h intervals with 2\u00a0mL of virus\u2010containing conditioned medium with 1.0\u00a0\u00b5g\u00b7mL\u22121 polybrene (Merck Millipore). Twenty\u2010four hours later, the infected cells were cultured in completed medium including suitable concentrations of puromycin for 3\u00a0days. Then, selected cells were utilized for the experiments.To obtain retroviruses, MSCV\u2010ires\u2010GFP or MSCV\u2010ires\u2010Puro encoding the cDNA of the indicated proteins was transfected into HEK293T cells against murine MSK1/2 and human K\u2010Ras gene products were inserted into pSUPER\u2010retro\u2010puro retroviral plasmid . The sequences of oligonucleotides used for constructing the shRNA retroviral vector against each gene product were described as below: For sh\u2010mMSK1#1; GATCCCCGTGATTTACCAGAGAGAAATTCAAGAGATTTCTCTCTGGTAAATCACTTTTTA and AGCTTAAAAAGTGATTTACCAGAGAGAAATCTCTTGAATTTCTCTCTGGTAAATCACGGG, For sh\u2010mMSK1#2; GATCCCCGCCAATACTCAGAAAGAAATTCAAGAGATTTCTTTCTGAGTATTGGCTTTTTA and AGCTTAAAAAGCCAATACTCAGAAAGAAATCTCTTGAATTTCTTTCTGAGTATTGGCGGG, For sh\u2010MSK2#1; GATCCCCGCGGAGAGCTATTGGAACATTCAAGAGATGTTCCAATAGCTCTCCGCTTTTTA and AGCTTAAAAAGCGGAGAGCTATTGGAACATCTCTTGAATGTTCCAATAGCTCTCCGCGGG, For sh\u2010MSK2#2; GATCCCCGGGCATGAGGAGAAGGTGATTCAAGAGATCACCTTCTCCTCATGCCCTTTTTA and AGCTTAAAAAGGGCATGAGGAGAAGGTGATCTCTTGAATCACCTTCTCCTCATGCCCGGG, For sh\u2010K\u2010Ras#1; GATCCCCCGAATATGATCCAACAATATTCAAGAGATATTGTTGGATCATATTCGTTTTTA and AGCTTAAAAACGAATATGATCCAACAATATCTCTTGAATATTGTTGGATCATATTCGGGG, For sh\u2010K\u2010Ras#2; GATCCCCGGACGAATATGATCCAACATTCAAGAGATGTTGGATCATATTCGTCCTTTTTA and AGCTTAAAAAGGACGAATATGATCCAACATCTCTTGAATGTTGGATCATATTCGTCCGGG.Infection with retroviruses, including these shRNA, into NIH\u20103T3 cells was performed at the same time with a retrovirus harboring oncogenic Ras.2.6\u22121 TNF\u03b1 for 16\u00a0h. Cells were lysed with passive lysis buffer , and the activity of expressed luciferase was measured by using a Luciferase Assay kit (Promega). The specific activity of luciferase was calculated by normalizing with protein concentration of each cell lysate, and data were shown in the graph as relative amount. To observe the effect of each kinase inhibitor on the NF\u2010\u03baB activation, cells were treated with the inhibitors 15\u00a0min prior to stimulation with TNF\u03b1. To analyze the transcriptional activation of NF\u2010\u03baB, parental NIH\u20103T3 cells were transfected with pFR\u2010luciferase containing the sequence of a GAL4\u2010responsive element with the indicated combination of plasmids. Cells were cultured in serum\u2010free DMEM for 24\u00a0h, and then, the cells were harvested, and luciferase activity was assayed as described above.KF\u20108 cells infected with indicated retroviruses were re\u2010plated on a 24\u2010well plate and then cultured in serum\u2010starved medium for 24\u00a0h. Then, the cells were stimulated with 10\u00a0ng\u00b7mL2.7et al., m HEPES\u2013KOH pH7.5 including 1\u00a0\u03bcg\u00b7\u03bcL\u22121 poly\u2010dIdC for 30\u00a0min at 25\u00a0\u00b0C. Then, the binding mixtures were pulled down using streptavidin\u2010conjugated agarose . The samples were analyzed by immunoblotting analysis as described below.Nuclear extracts were prepared as reported previously supplemented with protease inhibitors. FLAG\u2010MSK1 and FLAG\u2010PKAc were purified by immunoprecipitation using an M2 antibody attached to protein G\u2010sepharose , and they were utilized for in\u00a0vitro kinase assays. For these assays, purified kinases were incubated with 2\u00a0\u03bcg GST\u2010p65/RelA and 100\u00a0\u03bcm ATP for 30\u00a0min at 30\u00a0\u00b0C. The reaction was terminated by the addition of Laemmli sample buffer, and then, phosphorylated samples were resolved by SDS/PAGE and immunoblotting analysis with an anti\u2010phospho p65/RelA (S276) antibody .HEK293T cells were transfected with the indicated combinations of plasmids harboring cDNA of FLAG\u2010MSK1, FLAG\u2010PKAc, or H\u2010Ras (G12V). Twenty\u2010four hours later, the transfected cells were cultured in serum\u2010free DMEM for 24\u00a0h. Then, cells were harvested, and cell lysates were prepared with Nonidet P\u201040 (NP\u201040) lysis buffer (50\u00a0m2.9m sodium phosphate (pH 7.2), 150\u00a0mm NaCl, 3\u00a0mm MgCl2, 2\u00a0mm EDTA, 1% NP\u201040, 1% sodium deoxycholate, 0.2\u00a0U\u00b7mL\u22121 aprotinin, and phosphatase inhibitors] and briefly sonicated on ice. Then, debris were removed by sedimentation in a microcentrifuge at 16\u00a0400\u00a0g for 10\u00a0min, and cleared cell lysates were harvested and mixed with Laemmli sample buffer. Twenty\u2010five microgram of protein of whole cell lysates was loaded in each lane of an SDS\u2010polyacrylamide gel, and each protein was separated by the electrophoresis. Then, the separated proteins were transferred onto a polyvinylidene difluoride membrane (Merck Millipore). Proteins were visualized by immunoblotting analysis with the indicated antibodies and a chemical luminescence reagent, ECL . In some case such as the in\u00a0vitro kinase assay, cells were lysed with NP\u201040 lysis buffer as described above.For usual immunoblotting analysis, cells were lysed with RIPA buffer without SDS [10\u00a0m2.10m dNTPs, and 5\u00a0pmol oligo (dT)20 primer , and then, cDNA synthesis was performed for 60\u00a0min at 42\u00a0\u00b0C. The reaction was terminated by heating at 95\u00a0\u00b0C for 5\u00a0min and diluted with 80\u00a0\u03bcL TE buffer. One microlitre of synthesized cDNA was used for quantitative PCR in a 20\u00a0\u03bcL volume with the KAPA SYBR\u00ae FAST qPCR Kit , and the reaction was analyzed by a LightCycler 96 . The PCR primer sequences used are shown in Table Total RNA was extracted using TRIzol . To synthesize the single\u2010strand cDNA, 2\u00a0\u03bcg of total RNA was added into 20\u00a0\u03bcL reaction mixture including 100\u00a0units ReverTra Ace, 1\u00a0m2.114\u00a0cells per 35\u2010mm\u2010diameter dish, and grown for 2\u20133\u00a0weeks. The visible colonies showing a diameter of 1.0\u00a0mm or more were counted using nih imagej software, freely provided from Dr. Wayne Rasband in NIH (https://imagej.nih.gov/ij/), and results were shown in the graph.For the colony formation assay, infected NIH\u20103T3 cells were seeded onto soft agar at 1\u00a0\u00d7\u00a0102.12To detect K\u2010Ras mutations by quantitative PCR for the human colorectal tumor samples, and perform further experiments using them, we obtained informed consent forms from each patient. All experiments were undertaken with the understanding and written consent of each subject. The methodologies for experiments conformed to the standards set by the Declaration of Helsinki. The methodologies of current study were approved by the ethics committee in Jichi medical university.2.13et al., From tumor tissues of each patient, genomic DNA was extracted using a NucleoSpin Tissue XS Genomic DNA Purification kit in accordance with the manufacturer's instructions. The prepared genomic DNA was utilized for quantitative PCR analysis to detect K\u2010Ras mutations. Hydrolysis probes were designed in accordance with a previous report patients, and samples were fixed with formaldehyde. Among them, seven samples were selected for further analysis, because the tumor tissues in these samples harbored an oncogenic point mutation in exon 2 of the K\u2010Ras gene. The formalin\u2010fixed paraffin\u2010embedded sections were pretreated in a microwave oven for 15\u00a0min. First, H&E staining of each tumor tissue was performed to differentiate between tumor cell areas and normal tissues. Then, immunohistochemical staining was performed using anti\u2010p65/RelA or anti\u2010phospho p65/RelA (Ser\u2010276) as a primary antibody. As a secondary antibody, HRP\u2010conjugated anti\u2010rabbit IgG antibody (donkey) was used. Then, the sections were incubated with Envision and stained with DAB for the visualization of signals.2.15n\u00a0=\u00a03), and the results of calculations of independent t\u2010tests are shown.All experiments were performed individually three times, and representative data shown. In graphs, error bars indicate standard deviation drastically enhanced the expression of cyclin A and cyclin D1, while the expression of p27Kip1 was markedly diminished.Previously, we established KF\u20108 cells, a subline of NIH\u20103T3 cells stably harboring \u03baB\u2010responsive elements\u2010driven luciferase gene caused the hyperacceleration of NF\u2010\u03baB activation on TNF\u2010\u03b1\u2010induced signal transduction pathways for NF\u2010\u03baB activation. As shown in Fig. V on TNF\u20103.3Ink4a but not p27Kip1 suppressed TNF\u03b1\u2010induced NF\u2010\u03baB activation in HEK293 and HeLa cells and TNF\u03b1. Unexpectedly, we observed that the enforced expression of p16Ink4a could not inhibit the hyperactivation of NF\u2010\u03baB is a marker protein in the nucleolus, and some studies reported that NPM antagonizes p19ARF inhibitor p163.4m), U0126 (20\u00a0\u03bcm), and SP600125 (50\u00a0\u03bcm), slightly suppressed the hyperactivation of NF\u2010\u03baB, suggesting the partial involvement of the PI\u20103 kinase, ERK, and JNK pathways. Mitsushita et al. (m), an inhibitor for NOX1, exhibited slight inhibitory effect on NF\u2010\u03baB. Notably, treatment with 10\u00a0\u03bcm of SB203580, an inhibitor of p38 MAP kinase, exhibited drastic inhibitory effects on the oncogenic hyperactivation of NF\u2010\u03baB. These observations suggested the functional involvement of p38 MAP kinase in the H\u2010Ras (G12V)\u2010provoked oncogenic hyperactivation of NF\u2010\u03baB. As shown in Fig. m of SB203580 effectively suppressed the hyperactivation of NF\u2010\u03baB induced by co\u2010stimulation with B\u2010Raf (V600E) and TNF\u03b1 and MSK1, a downstream protein kinase of p38 MAP kinase, were reported to be protein kinases for the phosphorylation of p65/RelA at Ser\u2010276 , which catalyzes monomethylation of p65/RelA at Lys\u2010310 and suppresses the expression of NF\u2010\u03baB target genes. Additionally, several studies reported the importance of the phosphorylation of p65/RelA at Ser\u2010536 for the nuclear translocation or transcriptional activation of NF\u2010\u03baB KF\u20108 cells were infected with control retroviruses or retroviruses harboring K\u2010Ras (G12V) or c\u2010Myc. After puromycin selection, cells were stimulated with 10 \u03bcg/ml TNFa for indicated periods. Then, cells were lysed, and the luciferase assay was performed. (B) Using KF\u20108 cells infected with control retroviruses or retroviruses harboring K\u2010Ras mutants including G12V, K117N and A146T, the luciferase assay was performed as shown in (A). In the graph, error bars\u00a0=\u00a0SD .Fig. S2. (A) HEK293T cells were transfected with a luciferase vector including \u03baB\u2010responsive element with/without FLAG\u2010PKAc, a catalytic subunit of PKA. Then, using the transfected cells, luciferase assays were performed. In graph, error bars indicate standard deviation , and the results of calculations of independent t\u2010tests are shown (*P\u00a0<\u00a00.005). (B) The expression of FLAG\u2010MSK1 and H\u2010Ras (G12V) in cells analyzed for in vitro kinase assay was analyzed by immunoblotting analysis using antibodies against FLAG\u2010tag (M2) and H\u2010Ras. (C) KF\u20108 cells were infected with retroviruses harboring scrambled short\u2010hairpin RNA, sh\u2010MSK1 (1A or 1B) or sh\u2010MSK2 (2A or 2B). After puromycin selection, cells were lysed, and immunoblotting analysis was performed to evaluate the knockdown efficiency of MSK1 and MSK2. \u03b2\u2010actin was used as a loading control.Fig. S3. (A) NIH\u20103T3 cells were infected with indicated retroviruses as described in Figure 5B. After puromycin selection, cells (1 \u00d7 104 cells) were seeded on soft agar media in 35 mm dishes. After 3 weeks, the number of colonies were counted and shown in graph as shown in (B). In the graph, error bars\u00a0=\u00a0SD . In the photograph, 500 \u03bcm scale bar was shown.Fig. S4. SW620, a colorectal cancer\u2010derived cell line was transfected with siRNAs against both MSK1 and MSK2.Fig. S5. Genomic DNA was extracted from tumor tissues of 30 colorectal cancer patients (designated as T001 to T030).Fig. S6. Utilizing paired samples of tumor and normal tissues from K\u2010Ras (+) patients, immunohistochemical analyses were performed to detect total p65/RelA and phosphorylated p65/RelA (Ser276).Fig. S7. Total RNA was extracted from normal and tumor tissues from each patient harboring KRas mutations.Click here for additional data file.Table S1. Primers used in quantitative PCR for NF\u2010kB target genes.Click here for additional data file.Table S2. Primers used for genotyping of KRas gene.Click here for additional data file."} +{"text": "MecAAC (E=S (1); Se (2); Te (3); L=PhC(NtBu)2; MecAAC=C(CH2)(CMe2)2N\u20102,6\u2010iPr2C6H3)) were synthesized from the reactions of silylene\u2013phosphinidene LSi\u2212P\u2212MecAAC (A) with elemental chalcogens. All the compounds reported herein have been characterized by multinuclear NMR, elemental analyses, LIFDI\u2010MS, and single\u2010crystal X\u2010ray diffraction techniques. Furthermore, the regeneration of silylene\u2013phosphinidene (A) was achieved from the reactions of 2\u20133 with L\u2032Al (L\u2032=HC{(CMe)}2). Theoretical studies on chalcogen\u2010bonded silicon phosphinidenes indicate that the Si\u2212E bond can be best represented as charge\u2010separated electron\u2010sharing \u03c3\u2010bonding interaction between [LSi\u2212P\u2212MecAAC]+ and E\u2212. The partial double\u2010bond character of Si\u2212E is attributed to significant hyperconjugative donation from the lone pair on E\u2212 to the Si\u2212N and Si\u2212P \u03c3*\u2010molecular orbitals.Chalcogen\u2010bonded silicon phosphinidenes LSi(E)\u2212P\u2212 A, Scheme\u2005B, Scheme\u2005C, Scheme\u2005CycAAC)SiCl2\u2192P\u2212Tip .CycAAC)SiCl2\u2192P\u2212Tip Si(P\u2212Tip)]2 reacts smoothly with elemental chalcogens to form the chalcogen\u2010bonded silicon phosphinidenes LSi(E)\u2212P\u2212MecAAC (E=S (1); Se (2); Te (3)).Phosphinidenes (R\u2212P) are phosphorus analogues of carbenes and nitrenes.2, which is the only example of a compound with a silicon\u2013chalcogen double bond and a phosphine functionality.MecAAC (E=S (1); Se (2); Te (3)) with phosphinidene functionality. Compounds 1\u20133 were characterized by single\u2010crystal X\u2010ray structural investigation and multinuclear NMR spectroscopy. An equimolar reaction of compound A with elemental sulfur and selenium at room temperature in toluene afforded compounds 1 and 2 in 68\u2009% and 75\u2009% yield, respectively \u2212P\u2212MecAAC (3) in 79\u2009% yield }2) at room temperature as well as at elevated temperature, but no chalcogen transfer occurred. Nonetheless, 2 and 3 react with L\u2032Al at 60\u2009\u00b0C, resulting in the formation of parent silylene\u2013phosphinidene ; Te ) with ph1\u20133 are thermally stable with melting points over 200\u2009\u00b0C, but they are sensitive towards moisture. They are fully characterized by NMR spectroscopy, elemental analyses, LIFDI\u2010MS, and X\u2010ray single\u2010crystal structure analysis. The 31P{1H}\u2005NMR spectra of 1\u20133 exhibit a singlet at \u03b4=+2.33, +9.37, and +21.67\u2005ppm, respectively, which are highfield shifted compared with the starting material, silylene\u2013phosphinidene (A). In the 29Si{1H}\u2005NMR spectra, 1\u20133 display doublets at \u03b4=+20.48 (1JSi\u2212P=107\u2005Hz), +16.96 (1JSi\u2212P=117\u2005Hz), and \u221210.94\u2005ppm (1JSi\u2212P=127\u2005Hz), respectively, due to the coupling with the phosphorus atom. The difference of the chemical shift between 1\u20133 and A in the 29Si{1H}\u2005NMR spectra is due to the different silicon oxidation states (+4 and +2). The 77Se{1H}\u2005NMR spectrum of 2 shows a doublet at \u03b4=\u2212286.13\u2005ppm (2JSe\u2212P=18\u2005Hz) due to the coupling with the phosphorus atom. For the same reason the 125Te{1H}\u2005NMR spectrum of 3 exhibits a doublet at \u03b4=\u2212835.45\u2005ppm (2JTe\u2212P=38\u2005Hz). The LIFDI mass spectra of 1\u20133 in toluene exhibit molecular\u2010ion peaks at 607.4, 655.3, and 705.4, respectively.Compounds 1\u20133 suitable for X\u2010ray structural analysis were obtained from toluene solution either at 0\u2009\u00b0C or at room temperature (for details see the Supporting Information). Compounds 1\u20133 crystallize in the monoclinic space group P21/c. All three structures are isostructural, whereas 1 and 2 are even isomorphous. Compound 3 crystallizes as a pseudo\u2010merohedral twin with two molecules in the asymmetric unit. As a representative for all the molecular structure of 1 is depicted in Figure\u20051, 2, and 3 are about 3\u2005pm shorter than in A 2}Si(S)N(SiMe3)2] and [{PhC(NtBu)2}Si(Se)N(SiMe3)2] (1.987(8) and 2.136(9)\u2005\u00c5, respectively).3 is similar to the one observed in [{PhC(NtBu)2}Si(Te)N(SiMe3)2], (2.3720(15)\u2005\u00c5).1\u20133 are well within the range of previous reported Si\u2212E double bond lengths.Single crystals of A as well as chalcogen\u2010bonded silicon phosphinidenes 1, 2, and 3.A is given in Figure\u2005S2.3a (Supporting Information). The ESP value in the direction of the lone pair at P is slightly higher than that in the direction of the lone pair at Si . This indicates a slightly higher nucleophilicity of the first than the second (Table\u2005S2.1). The occupancy of the P (1.943\u2005e) and Si (1.925\u2005e) lone pair in A are well corroborated with ESP values (Table\u2005S2.2). This is in accordance with our previous theoretical study on silylene\u2013phosphinidenes.Quantum\u2010mechanical calculations were performed at the M06/def2\u2010TZVPP//BP86/def2\u2010SVP level of theory to understand the electronic structure and reactivity of silylene\u2013phosphinidene A, the reactions of silylene\u2013phosphinidene A with chalcogens result in silicon bonded chalcogens in the phosphinidenes 1, 2, and 3. To account for this observation, we have calculated the reaction energies for the formation of chalcogen\u2010bonded silicon phosphinidenes 1, 2, and 3 as well as hypothetical chalcogen\u2010bonded phosphorus phosphinidenes 1\u2032, 2\u2032, and 3\u2032 . All the reaction energies are exothermic, and the energies become more positive with the descent from sulfur to tellurium. The reaction energies for the formation of silicon\u2010bonded chalcogen phosphinidenes 1 (\u2212107.3), 2 (\u221288.2), and 3 are much higher than chalcogen\u2010bonded phosphorus phosphinidenes 1\u2032 (\u221277.0), 2\u2032 (\u221259.4), and 3\u2032 . Hence the formation of the P\u2212E bond is less favorable than the formation of a Si\u2212E bond. Note that our calculated reaction energies are comparable to the previously reported bond\u2010dissociation energies for Si=S (112.8), Si=Se (95.5), and Si=Te bonds in CH3Si(=E)OH compounds at the MP2/6\u2013311++G//MP2/6\u2013311+G level of theory.1\u20133, which is in accordance with the experimental observation that the regeneration of parent molecule A could not be achieved when 1 is treated with the L\u2032Al (L\u2032=HC{(CMe)}2) . The Si\u2212E bond lengths in 1 (2.019), 2 (2.160), and 3 (2.398\u2005\u00c5) in turn are close to those of previously reported Si=E double bond lengths.1 is comparable to those in the amidinate stabilized siladithiocarboxylate (2.030\u2005\u00c5),1\u20133 is in agreement with the Wiberg bond index of Si\u2212E bond , \u221236.1(2), and \u221232.0\u2005kcal\u2009mol\u22121 (3).The calculated geometrical parameters of 19), 2 2.0, and 3 MecAAC]+ and E\u2212. The second bonding interaction represents donor\u2013acceptor interaction between [LSi\u2212P\u2212MecAAC] and E. The best bonding representation is the one having the least value for orbital stabilization energy \u0394Eorb.Eorb for the charge separated electron sharing interaction in 1, 2, and 3 are \u2212215.8, \u2212189.1, and \u2212159.6\u2005kcal\u2009mol\u22121, respectively. The \u0394Eorb for the donor\u2013acceptor interaction in 1, 2, and 3 are \u2212265.2, \u2212214.7, and \u2212164.9\u2005kcal\u2009mol\u22121, respectively. Hence the best bonding description for the Si\u2212E bond in 1\u20133 is the charge\u2010separated electron\u2010sharing interaction between the fragments. However, the difference in the \u0394Eorb between the two bonding models in 3 is only 5.3\u2005kcal\u2009mol\u22121. Hence, the donor\u2013acceptor interaction also contributes towards the ground electronic structure of Si\u2212Te bond in 3.The EDA\u2010NOCV analysis was carried out to further shed light on the nature of the Si\u2212E bond. Two different bonding situations were analyzed, and the results are given in Table\u2005S2.5 (Supporting Information). The first bonding interaction represents charge separated electron\u2010sharing interaction between , wR2=0.0973 , res. density peaks: 0.324 to \u22120.251\u2005e\u2009\u00c5\u22123; Crystal data for 2: C35H54N3PSeSi, rM=654.83\u2005g\u2009mol\u22121, 0.351\u00d70.257\u00d70.145\u2005mm, monoclinic, P21/c, a=13.903(2), b=10.077(2), c=26.651(3)\u2005\u00c5, \u03b2=102.18(3)\u00b0, V=3649.8(11)\u2005\u00c53, Z=4, \u03bc (Mo K\u03b1)=1.131\u2005mm\u22121, \u03b8max=28.34\u00b0, 125\u2009128 reflections measured, 9105 independent (int=R0.0424), R1=0.0252 [I>2\u03c3(I)], wR2=0.0640 , res. density peaks: 0.406 to \u22120.182 e \u00c5\u22123; Crystal data for 3: C35H54N3PTeSi, rM=703.47\u2005g\u2009mol\u22121, 0.254\u00d70.119\u00d70.082\u2005mm, monoclinic, P21/c, a=35.937(3), b=9.237(2), c=23.498(3)\u2005\u00c5, \u03b2=109.09(2)\u00b0, V=7371(2)\u2005\u00c53, Z=8, \u03bc (Mo K\u03b1)=0.909\u2005mm\u22121, \u03b8max=25.32\u00b0, 120\u2009742 reflections measured, 13\u2009421 independent (int=R0.0522), R1=0.0259 [I>2\u03c3(I)], wR2=0.0500 , res. density peaks: 0.570 to \u22120.559\u2005e\u2009\u00c5\u22123.The datasets were collected on an Incoatec Mo Microsource1), 1891854 (2), and 1891855 (3)1891853 should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "II complex and a CuII open-cube complex were synthesized using the tridentate Schiff base ligand, {1-[1-(pyrid\u00adyl)(2-eth\u00adoxy\u00adethyl\u00adidene)]-2-[(pyridin-3-yl)carbon\u00adyl]}hydrazine and copper(II) acetate and their mol\u00adecular and crystal structures determined.A linear polymeric Cu II complexes [Cu(C14H13N4O2)Cl]n, I, and [Cu4(C8H10NO2)4Cl4]n, II, have been synthesized. In the structure of the mononuclear complex I, each ligand is coordinated to two metal centers. The basal plane around the CuII cation is formed by one chloride anion, one oxygen atom, one imino and one pyridine nitro\u00adgen atom. The apical position of the distorted square-pyramidal geometry is occupied by a pyridine nitro\u00adgen atom from a neighbouring unit, leading to infinite one-dimensional polymeric chains along the b-axis direction. Each chain is connected to adjacent chains by inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efCl inter\u00adactions, leading to a three-dimensional network structure. The tetra\u00adnuclear complex II lies about a crystallographic inversion centre and has one core in which two CuII metal centers are mutually inter\u00adconnected via two enolato oxygen atoms while the other two CuII cations are linked by a chloride anion and an enolato oxygen. An open-cube structure is generated in which the two open-cube units, with seven vertices each, share a side composed of two CuII ions bridged by two enolato oxygen atoms acting in a \u03bc3-mode. The CuII atoms in each of the two CuO3NCl units are connected by one \u03bc2-O and two \u03bc3-O atoms from deprotonated hydroxyl groups and one chloride anion to the three other CuII centres. Each of the penta\u00adcoordinated CuII cations has a distorted NO3Cl square-pyramidal environment. The CuII atoms in each of the two CuO2NCl2 units are connected by \u03bc2-O and \u03bc3-O atoms from deprotonated alcohol hy\u00addroxy groups and one chloride anion to two other CuII ions. Each of the penta\u00adcoordinated CuII cations has a distorted NO2Cl2 square-pyramidal environment. In the crystal, a series of intra\u00admolecular C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds are observed in each tetra\u00adnuclear monomeric unit, which is connected to four tetra\u00adnuclear monomeric units by inter\u00admolecular C\u2014H\u22efO hydrogen bonds, thus forming a planar two-dimensional structure in the (Two Cu It has been shown that the presence of metal ions promotes the hydrolysis of the ester function of the picolinic ester ions. These ligands then coordinate to the copper(II) cations to yield the two complexes that are reported here.Picolinic acid esters methanol] and a condensation product [({1-[1-eth\u00adoxy-1-(pyridin-2-yl)methyl\u00adene]}-2-(oxonicotin\u00adyl))hydrazine]. It has been shown (Papaefsta\u00adthiou 14H13N4O2)]n, I, the repeat unit of which is shown in \u2005\u00c5, an O16 atom with a Cu1\u2014O16 distance of 1.9808\u2005(15)\u2005\u00c5 and the N11 and N22 atoms from the same ligand with a Cu\u2014N distances of 1.9437\u2005(17) and 2.0444\u2005(17)\u2005\u00c5 \u201379.40\u2005(7)\u00b0, which are slightly smaller than those found in similar compounds \u00b0 and 146.17\u2005(6)\u00b0 methano\u00adlate ligand coordinating to each Cu atom through its imine nitro\u00adgen atom and its alcoholate oxygen atom, forming five-membered chelate rings . Intra\u00admolecular C\u2014H\u22efO contacts are also found \u2212x\u00a0+\u00a0y\u00a0\u2212\u00a0z\u00a0+\u00a0i, Cu2i atoms, \u03bc3-bridging atoms O26, O26i, a \u03bc2-bridging O15i atom and a \u03bc2-bridging Cl3 ion. The result is a is a distorted open-cube, defined as a distorted cube missing one corner. This can be seen by considering that the range of Cu\u2014O\u2014Cu angles is [99.76\u2005(6)\u2013102.98\u2005(6)\u00b0] and the Cu1\u2014Cl3\u2014Cu2i angle is 84.39\u2005(2)\u00b0. These differ extensively from the 90\u00b0 angles of an ideal cube. The two Cu3O3Cl open-cubes are joined by a perfectly rectangular side defined by the Cu1, O26, and Cui, O26i atoms. The values of the two different lengths of the edges of the rectangular sides are 2.4280\u2005(14) and 1.9684\u2005(13)\u2005\u00c5. The other faces of the two open-cubes are irregular with different distances i.e. Cu1\u2014O26i = 2.4280\u2005(14)\u2005\u00c5, Cu2\u2014O26 = 1.9707\u2005(14)\u2005\u00c5, Cu1\u2014Cl3 = 2.2181\u2005(6)\u2005\u00c5 and Cu2\u2014Cl3 = 2.8134\u2005(6)\u2005\u00c5. The Cu1 (Cu1i) atoms in each of the two CuO3NCl units are connected by one \u03bc2-O and two \u03bc3-O atoms from the deprotonated hydroxyl groups and one chloride ion to three other CuII cations. In the CuO2NCl2 units, the Cu2 (Cu2i) atoms are linked to one \u03bc2-O and one \u03bc3-O atoms from a deprotonated hydroxyl groups and one chloride ion to two other CuII cations with Cu1\u2014Cu2 and Cu1\u2014Cu2i distances of approximately 3.012 and 3.408\u2005\u00c5, respectively. These are in good agreement with literature values , 1.9684\u2005(13), 1.9707\u2005(14)\u2005\u00c5, respectively, while Cu1\u2014O15 and Cu2\u2014O15 are 1.9170\u2005(13) and 1.9324\u2005(13)\u2005\u00c5, respectively \u00b0], O26\u2014Cu1\u2014N10 [156.02\u2005(7)\u00b0], O26\u2014Cu2\u2014Cl4 [170.05\u2005(5)\u00b0] and O15\u2014Cu2\u2014N21 [157.61\u2005(7)\u00b0] \u00b0 are different from those of ideal cube. This bridging angle is also smaller than those reported for similar complexes 2Cl2](DMF) where qsalBr = 8-amino\u00adquinoline with 5-bromo-salicyl\u00adaldehyde . These contacts combine with the C\u2014H\u22efO hydrogen bonds to stack the mol\u00adecules in a three-dimensional network along the a-axis direction ]methyl\u00adenehydrazine yielded no hits, indicating that compound I is reasonably unique. However, a search for eth\u00adoxy(pyridin-2-yl)methano\u00adlate, the ligand found in II gave ten hits, although none of these was closely related to II. The matches included the CuII complexes HAXBEN meth\u00adyl]pyridin, has a central pyridine ring that is substituted by 1-eth\u00adoxy-1-hy\u00addroxy-1-(pyridin-2-yl)methyl fragments in the 2- and 6-positions.A search of the CSD database 2\u00b7H2O in 5\u2005ml of ethanol was added at room temperature. The initial yellow solution immediately turned deep blue and was stirred under reflux for 2\u2005h. The mixture was filtered and the solution evaporated to near dryness. The solid was isolated by filtration and recrystallized from a minimum of ethanol. On standing for five days, two types of crystals suitable for X-ray analysis were formed, light-yellow blocks of I and light-green plates of II.To a solution of 2-pyridine carbaldehyde in 30\u2005ml of ethanol was added a solution of nicotinic hydrazide in 10\u2005ml of ethanol. The mixture was stirred for 5\u2005min. A solution of Cu: 2982, 1628, 1583, 1423, 1343, 1245, 941, 816, 630. For II: analysis calculated: C16H20N2Cl2O4Cu2: C, 38.26; H, 4.01; N, 5.58; Cl; 14.12. Found: C, 38.23; H, 3.98; N, 5.55; Cl; 14.08. IR : 2982, 1585, 1423, 1243, 1145, 940, 812.For d(C\u2014H) = 0.93\u2005\u00c5 for aromatic, d(C\u2014H) = 0.97\u2005\u00c5 for methyl\u00adene and d(C\u2014H) = 0.98\u2005\u00c5 for methine H atoms with Uiso(H) = 1.2Ueq(C) and d(C\u2014H) = 0.96\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for methyl H atoms. One reflection with Fo <<< Fc that was likely to have been affected by the beamstop was omitted from the final refinement cycles.Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989019008922/sj5575sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989019008922/sj5575Isup4.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019008922/sj5575IIsup5.hklStructure factors: contains datablock(s) II. DOI: 1935842, 1935841CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Hirshfeld surface analysis of both com\u00adpounds have been carried out.The mol\u00adecular and crystal structures of \u03bc-aqua-\u03ba 2O:O-di-\u03bc-di\u00adphenyl\u00adacetato-\u03ba4O:O\u2032-bis\u00ad[(di\u00adphenyl\u00adacetato-\u03baO)bis\u00ad(pyridine-\u03baN)nickel(II)], [Ni2(C14H11O2)4(C5H5N)4(H2O)] (1) and \u03bc-aqua-\u03ba2O:O-di-\u03bc-di\u00adphenyl\u00adacetato-\u03ba4O:O\u2032-bis\u00ad[(di\u00adphenyl\u00adacetato-\u03baO)nickel(II)]\u2013aceto\u00adnitrile\u2013di\u00adphenyl\u00adacetic acid (1/2.5/1), [Ni2(C14H11O2)4(C10H8N2)2(H2O)]\u00b72.5CH3CN\u00b7C14H12O2 (2), the com\u00adplex units are stabilized by a variety of intra- and inter\u00admolecular hydrogen bonds, as well as C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 contacts between the aromatic systems of the pyridine, dipyridyl and di\u00adphenyl\u00adacetate ligands. Despite the fact that the di\u00adphenyl\u00adacetate ligand is sterically bulky, this does not inter\u00adfere with the formation of the described aqua-bridged dimeric core, even with a 2,2\u2032-bi\u00adpyridine ligand, which has a strong chelating effect.In the crystal structures of the title com\u00adpounds, namely \u03bc-aqua-\u03ba They belong to the type of aqua-bridged dinickel(II) carboxyl\u00adates with the general formula [MII2(\u03bc-H2O)(\u03bc-O2CR)2(O2CR)2nL] (n = 4 in the case of a monodentate ligand or 2 in the case of bidentate coordination), well known since the 1970s (for 1) or one 2,2\u2032-bi\u00adpyridine (Bipy) ligand (for 2) and one O atom from a bridging aqua ligand in an octa\u00adhedral geometry and 3.4826\u2005(5)\u2005\u00c5 (for 2). Each monodentate coordinated di\u00adphenyl\u00adacetate ligand is involved in the formation of an intra\u00admolecular hydrogen bond with a bridging water mol\u00adecule. Hydrogen-bond geometries are specified in Tables 1In the title binuclear com\u00adplexes, y Figs. 1 and 2 \u25b8.1, mol\u00adecules are combined into pairs connected by a centre of symmetry using offset face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions, in which coordinated pyridine ligands of each com\u00adplex are involved. These pairs, which result from the intermolecular hydrogen bonds between Py ligands and O atoms of carbonyl groups of di\u00adphenyl\u00adacetate ligands , form layered structures parallel to the (101) plane. The layers form a 3D supra\u00admolecular structure through inter\u00admolecular C\u2014H\u22ef\u03c0 contacts, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions between the phenyl substituents of di\u00adphenyl\u00adacetate ligands .In the crystal packing of com\u00adpound 2, each mol\u00adecule of the com\u00adplex is associated with two neighbouring mol\u00adecules via \u03c0\u2013\u03c0 stacking inter\u00adactions between Bipy ligands . As a result, polymer chains are formed along the , as well as N\u22efH inter\u00adactions involving aceto\u00adnitrile solvent mol\u00adecules .For com\u00adpound ds Fig.\u00a07. Other o1, close C\u22efC inter\u00adplanar contacts, responsible for \u03c0\u2013\u03c0 stacking inter\u00adactions between Bipy ligands, are displayed as patches of combined blue and red triangles on the surface over shape index fragment and no binuclear structures were found. A survey of the CSD reveals 46 related structures of NiII carboxyl\u00adates with a similar structure fragment.A search in the Cambridge Structural Database , in aceto\u00adnitrile (40\u2005ml) was added to a solution of di\u00adphenyl\u00adacetic acid in 10\u2005ml aceto\u00adnitrile. After full conversion of hellyerite, pyridine was added and the solution was refluxed for 15\u2005min. The resulting pale-green\u2013blue solution was cooled to room temperature and filtered. After a few days, blue crystals of 1 were collected by filtration (yield \u223c70%).A suspension of synthetic hellyerite, NiCO2 was carried out in a similar manner to the synthesis of com\u00adpound 1, but 2,2\u2032-bi\u00adpyridine was added instead of pyridine. The resulting blue solution was cooled to room temperature and filtered. After a few days, blue crystals of 2 were collected by filtration (yield \u223c60%).The synthesis of com\u00adpound Uiso(H) = 1.2Ueq(C) for the tertiary C atoms, C\u2014H = 0.93\u2005\u00c5 and Uiso(H) = 1.2Ueq(C) for aromatic C atoms, and C\u2014H = 0.96\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019014063/su5519sup1.cifCrystal structure: contains datablock(s) global, 1, 2. DOI: 10.1107/S2056989019014063/su55191sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989019014063/su55192sup3.hklStructure factors: contains datablock(s) 2. DOI: 1959435, 1959436, 1959435, 1959436CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 10.1038/s41598-018-30356-2, published online 10 August 2018Correction to: This Article contains errors in the Methods under subheading \u2018IBV protection experiment 2\u2019.7 EID50 of rNDV and rNDV/codon optimized-S, respectively, via oculanasal route. Five chickens of group three were inoculated with 10 recommended doses of a commercial live attenuated Mass-type IBV vaccine via oculanasal route and chickens of group four were inoculated with PBS.\u201d\u201cA total of twenty 4-week-old SPF chickens were divided into four groups of five each. Five chickens of groups one and two were inoculated with 10should read:7 EID50 of rNDV/codon optimized-S and 10 recommended doses of a commercial live attenuated Mass-type IBV vaccine via oculanasal route, respectively. Chickens of group three were inoculated with PBS. Samples collected from five non-vaccinated SPF chickens involved in another IBV protection study also were used as control.\u201d\u201cA total of fifteen 4-week-old SPF chickens were divided into three groups of five each. Five chickens of groups one and two were inoculated with 10Furthermore, within the Methods under subheading \u2018Quantitative reverse transcription-polymerase chain reaction (RT-qPCR)\u2019\u201cForty cycles of PCR at 95\u2009\u00b0C for 10\u2009s (denaturation), 58\u2009\u00b0C for 20\u2009s , and 72\u2009\u00b0C for 30\u2009s (elongation) followed by melting curve analysis that consisted of 95\u2009\u00b0C for 5\u2009s and 65\u2009\u00b0C for 60\u2009s.\u201dshould read:\u201cThe thermal conditions of 50\u2009\u00b0C for 10\u2009min and 95\u2009\u00b0C for 10\u2009min, then forty-five cycles of PCR at 95\u2009\u00b0C for 10\u2009s (denaturation), 58\u2009\u00b0C for 20\u2009s followed by 95\u2009\u00b0C for 5\u2009s and melting curve analysis that consisted of 65\u2009\u00b0C to 95\u2009\u00b0C for 5\u2009s was carried out to amplify the 150\u2009nt \u2013 N gene fragments.\u201d"} +{"text": "A new symmetrical thio\u00adcarbonohydrazone derivative with two similar benzoyl\u00adthio\u00adureido functional groups has been prepared and characterized. 18H18N4O2S2, which consists of two benzoyl\u00adthio\u00adureido moieties connected by an ethyl\u00adene chain. The asymmetric unit consists of one half of the mol\u00adecule, the complete mol\u00adecule being generated by crystallographic inversion symmetry. Both thio\u00adurea moieties are in a trans conformation. An intra\u00admolecular N\u2014H\u22efO hydrogen bond occurs. In the crystal, C\u2014H\u22efS and C\u2014H\u22efO hydrogen bonds link the molecules, forming layers parallel to the ac plane.The reaction of benzoyl chloride and ethyl\u00adendi\u00adamine in the presence of potassium thio\u00adcyanate yielded a white solid, C CuII, NiII and CoII) from acidic media. Thio\u00adurea derivatives have also been shown to possess anti\u00adbacterial, anti\u00adfungal, anti\u00adtubercular, anti\u00adthyroid and insecticidal properties \u2005\u00c5] and O1\u2014C3 [1.2209\u2005(16)\u2005\u00c5] distances indicate that these correspond to double bonds and are comparable to those observed for 1,2-bis\u00ad(N-benzoyl\u00adthio\u00adureido)benzene [1.6574\u2005(18)\u2005\u00c5 for S\u2014C and 1.222\u2005(2)\u2005\u00c5 for O7\u2014C16] rings plane. Mol\u00adecular layers running almost parallel to the ac plane are formed by inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efS inter\u00adactions \u2005\u00c5; Cg1 is the centroid of ring C4\u2013C9; symmetry code: (iv) \u2212x\u00a0+\u00a01, y\u00a0+\u00a0z\u00a0+\u00a0In the crystal, the mol\u00adecules, which feature an overall Z-form, have both halves roughly parallel to the s Table\u00a01. These let al., 2016et al., 2016trans with respect to the thiono S atom across the C\u2014N bond. The 1-benzoyl-3-ethyl\u00adthio\u00adurea fragment adopts a cis conformation with respect to the thiono S atom across the respective C\u2014N bond. Six structures in which the spacer is different from the spacer in the symmetrical bis\u00ad(thio\u00adureido) mol\u00adecule studied here appear in the literature. The angles between the phenyl rings are: 63.1\u00b0 for DAVHOZ was added and a white solid appeared after a few minutes. The compound was filtered off, washed with 3 \u00d7 50\u2005ml of water and dried under vacuum. The solid product was washed with water and purified by recrystallization from an ethanol/di\u00adchloro\u00admethane mixture (1:1 v/v). 12.3\u2005g of the title compound were obtained (yield 88.5%). A small qu\u00adantity of powder was recrystallized from 5\u2005ml of DMF. Colourless single crystals suitable to XRD grew within six days.All purchased chemicals and solvents were of reagent grade and were used without further purification. Melting points were determined with a B\u00fcchi 570 melting-point apparatus and were uncorrected. To a mixture of 7.02\u2005g (72\u2005mmol) of potassium thio\u00adcyanate and 100\u2005ml of acetone was added dropwise a solution of 10.116\u2005g (72\u2005mmol) of benzoyl chloride in 50\u2005ml of acetone. The resulting mixture was stirred under reflux for 1\u2005h and cooled to room temperature. A solution of 2.2\u2005g (36.6\u2005mmol) of 1,2-ethyl\u00adenedi\u00adamine in 20\u2005ml of acetone was added. The yellow solution obtained was stirred at room temperature during 2\u2005h. Hydro\u00adchloric acid (0.1\u2005arH) or 0.97\u2005\u00c5 (CH2). The NH H atoms were located in a difference Fourier map and freely refined.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901900495X/vm2216sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901900495X/vm2216Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901900495X/vm2216Isup3.cmlSupporting information file. DOI: 1909438CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Conventional hydrogen-bonding inter\u00adactions lead to supra\u00admolecular tapes in the crystal.The 4-pyridyl residues lie to either side of the central, planar C 14H14N4O2\u00b72C7H6O2, are described. The oxalamide mol\u00adecule has a (+)-anti\u00adperiplanar conformation with the 4-pyridyl residues lying to either side of the central, almost planar C2N2O2 chromophore (r.m.s. deviation = 0.0555\u2005\u00c5). The benzoic acid mol\u00adecules have equivalent, close to planar conformations [C6/CO2 dihedral angle = 6.33\u2005(14) and 3.43\u2005(10)\u00b0]. The formation of hy\u00addroxy-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonds between the benzoic acid mol\u00adecules and the pyridyl residues of the di\u00adamide leads to a three-mol\u00adecule aggregate. Centrosymmetrically related aggregates assemble into a six-mol\u00adecule aggregate via amide-N\u2014H\u22efO(amide) hydrogen bonds through a 10-membered {\u22efHNC2O}2 synthon. These are linked into a supra\u00admolecular tape via amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds and 22-membered {\u22efHOCO\u22efNC4NH}2 synthons. The contacts between tapes to consolidate the three-dimensional architecture are of the type methyl\u00adene-C\u2014H\u22efO(amide) and pyridyl-C\u2014H\u22efO(carbon\u00adyl). These inter\u00adactions are largely electrostatic in nature. Additional non-covalent contacts are identified from an analysis of the calculated Hirshfeld surfaces.The crystal and mol\u00adecular structures of the title 1:2 co-crystal, C Indeed, in early studies on crystal engineering, the combination of bifunctional nLH2 with di\u00adcarb\u00adoxy\u00adlic acids such as bis\u00ad(carb\u00adoxy\u00admeth\u00adyl)oxalamide -antiperiplanar conformation where the 4-pyridyl residues lie to either side of the central C2N2O2 chromophore. The six atoms comprising the central residue are close to co-planar with their r.m.s. deviation equal to 0.0555\u2005\u00c5, with the maximum deviations to either side of the plane being 0.0719\u2005(5) and 0.0642\u2005(5)\u2005\u00c5 for the N2 and O2 atoms, respectively; the C6 and C9 atoms lie 0.1908\u2005(14) and 0.0621\u2005(14)\u2005\u00c5 out of and to one side of the plane (towards the N2 atom), respectively. The N1- and N4-pyridyl rings form dihedral angles of 86.00\u2005(3) and 83.34\u2005(2)\u00b0, respectively, with the plane through the C2N2O2 atoms, so are close to perpendicular to the central plane. The dihedral angle between the pyridyl rings is 33.60\u2005(5)\u00b0, indicating a splayed disposition as each pyridyl ring is folded away from the rest of the mol\u00adecule. The carbonyl groups are anti and the mol\u00adecule features intra\u00admolecular amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds that complete S(5) loops, Table\u00a01The mol\u00adecular structures of the three constituents comprising the crystallographic asymmetric unit of (I)2 group being 6.33\u2005(14) and 3.43\u2005(10)\u00b0 for the O3- and O5-benzoic acid mol\u00adecules, respectively. As expected, the C15\u2014O3(carbon\u00adyl) bond length of 1.2162\u2005(13)\u2005\u00c5 is significantly shorter than the C15\u2014O4(hy\u00addroxy) bond of 1.3197\u2005(13)\u2005\u00c5; the bonds of the O5-benzoic acid follow the same trend with C22\u2014O5 of 1.2237\u2005(13)\u2005\u00c5 compared with C22\u2014O6 of 1.3084\u2005(13)\u2005\u00c5.There are two independent benzoic acid mol\u00adecules in (I)via amide-N\u2014H\u22efO(amide) hydrogen bonding, which leads to a centrosymmetric ten-membered {\u22efHNC2O}2 synthon. The second amide forms an amide-N\u2014H\u22efO(carbon\u00adyl) bond with the result of that adjacent six-mol\u00adecule aggregates are connected into a supra\u00admolecular tape via 22-membered {\u22efHOCO\u22efNC4NH}2 synthons, Fig.\u00a02a). The other notable contact within the tape is a pyridyl-C\u2014H\u22efO(carbon\u00adyl) inter\u00adaction, which cooperates with a hy\u00addroxy-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bond to form a seven-membered {\u22efOCOH\u22efNCH} pseudo-heterosynthon; no analogous inter\u00adaction is noted for the O5-benzoic acid. The supra\u00admolecular tapes are aligned along the c-axis direction and have a linear topology.As anti\u00adcipated from the chemical composition, significant conventional hydrogen bonding is noted in the crystal of (I)i.e. methyl\u00adene-C\u2014H\u22efO(amide) and pyridyl-C\u2014H\u22efO(carbon\u00adyl), the latter involving both pyridyl rings and each carbonyl-O atom, Table\u00a01b).The connections between chains leading to a three-dimensional architecture are of the type C\u2014H\u22efO, Crystal Explorer 17 hydrogen bonds was used as the input for calculations. A list of the short inter\u00adatomic contacts discussed below is given in Table\u00a02dnorm surfaces, Fig.\u00a034LH2 and BA mol\u00adecules, hereafter BA-I for the O3-containing mol\u00adecule and BA-II for the O5-mol\u00adecule, which indicate the presence of close contacts with distances shorter than the sum of the respective van der Waals radii , benzoic acid-O6\u2014H6O\u22efN4(pyrid\u00adyl), amide-N2\u2014H2N\u22efO2(amide) and amide-N3\u2014H3N\u22efO5(carbon\u00adyl) hydrogen-bonding inter\u00adactions exhibit the most intense red spots on the dnorm surfaces, suggestive of strong inter\u00adactions.The program a) and (b) [in the order of moderate intensity (m) to weak intensity (w)] were identified for C6\u2014H6B\u22efO1 (m), C12\u2014H12\u22efO3 (m), C2\u2014H2\u22efO5 (m) and C1\u2014H1\u22efO3 (w), Table\u00a01m), C28\u2014H28\u22efO1 (w), C9\u2014H9B\u22efC17 (w), C9\u2014H9B\u22efO3 (w), C13\u2014H13\u22efC1 (w), C12\u22efC21 (w) and C8\u22efC26 (w), Table\u00a02PLATON and (d).Other, relatively less intense red spots in Fig.\u00a034LH2, BA-I and BA-II mol\u00adecules through DFT-B3LYP/6-31G to further study the nature of the close contacts, Fig.\u00a04-\u2013H\u22efN and N\u2014H\u22efO hydrogen-bonding contacts that exhibited the most intense red spots on the dnorm map are highly electrostatic in nature, as evidenced from the intense electronegative (red) and electropositive (blue) regions on the Hirshfeld surfaces of the individual mol\u00adecules. Other regions are relatively pale, indicating the complementary role of the remaining contacts in sustaining the mol\u00adecular network in the crystal.The electrostatic potential mapping was performed on the individual i.e. the three-mol\u00adecule aggregate specified above, as well as its individual 4LH2, BA-I and BA-II components, Fig.\u00a054LH2 and benzoic acid in constructing the mol\u00adecular packing of the system, in contrast to the previously reported benzene monosolvate of 4LH2 i.e. the donor or acceptor atoms inter\u00adnal to the surface) and external contacts. In contrast, for H\u22efC/C\u22efH and H\u22efO/O\u22efH, the distributions are slightly inclined towards -C\u22efH- (15.8%) and -O\u22efH- (13.4%) as compared to the corresponding counterparts at 11.7 and 11.8%, respectively. A detailed analysis of the di + de distances shows that the closest H\u22efO/O\u22efH and H\u22efC/C\u22efH contacts of \u223c1.95\u2005\u00c5 and \u223c2.62\u2005\u00c5, respectively, occur at distances shorter than the sum of the respective van der Waals radii of 2.61 and 2.79\u2005\u00c5, while the H\u22efH (\u223c2.20\u2005\u00c5) and N\u22efH/ H\u22efN (2.80\u2005\u00c5) contacts are longer than the sum of van der Waals radii of 2.18 and 2.64\u2005\u00c5, respectively.The major surface contacts for (I)4LH2 mol\u00adecule also displays a shield-like profile with asymmetric spikes which upon further decomposition could be delineated into H\u22efH (38.0%), H\u22efO/O\u22efH (25.6%), H\u22efC/C\u22efH (21.4%) and H\u22efN/N\u22efH (9.9%) contacts. The H\u22efO/O\u22efH contact exhibits a forceps-like profile with the distribution inclined towards inter\u00adnal-H\u22efO-external (15.2%) as compared to inter\u00adnal-O\u22efH-external (10.4%), and both with tips at di + de \u223c1.94\u2005\u00c5 which is indicative of significant hydrogen bonding. Similarly, the asymmetric, needle-like profile for the H\u22efN/N\u22efH contact is inclined towards the inter\u00adnal-N\u22efH-external (9.0%) with the tip at di + de = \u223c1.6\u2005\u00c5, while the remaining 0.9% is attributed to the inter\u00adnal-H\u22efN-external contact with di + de of \u223c2.94\u2005\u00c5 . The H\u22efC/C\u22efH contacts are evenly distributed on both sides of the contacts with the di + de of \u223c2.64\u2005\u00c5 which is slightly shorter than the sum of van der Waals radii. On the other hand, the H\u22efH contacts have little direct influence in sustaining the mol\u00adecular packing as shown from the shortest di + de value of \u223c2.2\u2005\u00c5, which is longer than the sum of the van der Waals radii despite the prominent contributions these make to the overall surfaceThe i.e. 31.9% for BA-I cf. 38.7% for BA-II. The discrepancy in the distribution for BA-I is compensated by the increase in O\u22efC/C\u22efO and C\u22efC contacts with the distribution being 4.8 and 2.9%, respectively. The distribution for H\u22efC/C\u22efH (29.0 vs 29.1%), H\u22efO/O\u22efH (23.8 vs 24.2%) and H\u22efN (6.7 vs 5.7%) contacts is approximately the same in both BA-I and BA-II, except that the H\u22efC/C\u22efH distribution for BA-II is significantly more inclined towards inter\u00adnal-C\u22efH-external (20.5%) than the inter\u00adnal-H\u22efC-external (8.6%) in contrast to the relatively balanced distribution for BA-I 15.5% for inter\u00adnal-C\u22efH-external vs 13.5% for inter\u00adnal-H\u22efC-external. In BA-I, the di + de values for H\u22efO/O\u22efH, H\u22efC/C\u22efH and H\u22efN/N\u22efH at the tips are \u223c2.26\u20132.70, 2.62 and 1.64\u2005\u00c5, respectively, while the equivalent values for the analogous contacts for BA-II have tips at 2.02\u20132.56, 2.62\u20132.86 and 1.58\u2005\u00c5, respectively. Among these contact distances, the O\u22efH, H\u22efC/C\u22efH and H\u22efN for BA-I as well as H\u22efO/O\u22efH, H\u22efC and H\u22efN for BA-II are shorter than the sum of van der Waals radii. As expected, the minimum di + de value for the H\u22efH contacts is longer than the sum of van der Waals radii, even if it is the most dominant contact for each mol\u00adecule. The aforementioned data for BA-I and BA-II clearly distinguishes the independent mol\u00adecules.As for the pair of BA mol\u00adecules, both BA-I and BA-II possess similar, claw-like profiles which differ in the diffuse region, with the former being the characteristic of H\u22efH contacts while the latter is due to H\u22efC/C\u22efH inter\u00adactions. Qu\u00adanti\u00adtatively, differences mainly relate to the percentage contribution by H\u22efH contacts, CrystalExplorer17 hydrogen bonds complemented by a pair of pyridyl-C13\u2014H13\u22efC1(pyrid\u00adyl) inter\u00adactions between two oxamide mol\u00adecules led to the greatest inter\u00adaction energy (Etot) of \u221275.2\u2005kJ\u2005mol\u22121. This value is comparable to Etot of \u221271.7\u2005kJ\u2005mol\u22121 calculated for the classical eight-membered {\u22efHOCO}2 inter\u00adaction and pyridyl-C1\u2014H1\u22efO3(carbon\u00adyl) contacts, which combine to generate a seven-membered heterosynthon with Etot of \u221250.1\u2005kJ\u2005mol\u22121. A diminution in Etot is observed for the other pyridyl terminus, which only comprises a carb\u00adoxy\u00adlic-O6\u2014H6O\u22efN4(pyrid\u00adyl) hydrogen bond without a supporting pyridyl-C\u2014H\u22efO(carbon\u00adyl) inter\u00adaction, showing an energy of \u221243.9\u2005kJ\u2005mol\u22121 and ranked third strongest among all inter\u00adactions in (I)To assess the strength of the specified inter\u00adactions in the Hirshfeld surface analysis, the mol\u00adecules in (I)Etot of \u221236.3\u2005kJ\u2005mol\u22121 involves contributions from the amide-N3\u2014H3N\u22efO5(carb\u00adoxy\u00adlic acid) and phenyl-C28\u2014H28\u22efO1(amide) contacts. Other inter\u00adactions include the methyl\u00adene-C6\u2014H6B\u22efO1(amide) contacts (\u201325.1\u2005kJ\u2005mol\u22121), the combination of benzoic acid-C20\u2014H20\u22efC8(amide) and benzoic acid-C12\u22efC21(pyrid\u00adyl) (\u201321.0\u2005kJ\u2005mol\u22121), amide-C8\u22efC26(benzoic acid) (\u201319.4\u2005kJ\u2005mol\u22121), pyridyl-C2\u2014H2\u22efO5(carb\u00adoxy\u00adlic acid) (\u201315.6\u2005kJ\u2005mol\u22121), a combination of (pyridine meth\u00adyl)-C9\u2014H9B\u22efO3(carb\u00adoxy\u00adlic acid) and methyl\u00adene-C9\u2014H9B\u22efC17(benzoic acid) (\u221214.7\u2005kJ\u2005mol\u22121), as well as pyridyl-C12\u2014H12\u22efO3(carb\u00adoxy\u00adlic acid) (\u20138.3\u2005kJ\u2005mol\u22121). Some inconsistencies are observed between the calculated Etot and Hirshfeld surface analysis, particularly for C2\u2014H2\u22efO5 and C12\u2014H12\u22efO3. These inter\u00adactions can be considered weak even though they possess a relatively short contact distance compared to the sum of van der Waals radii, as indicated from the moderately intense red spots on the Hirshfeld surface. The contradiction could arise as a result of the relatively high repulsion terms, which weaken the inter\u00adaction energy.The next highest inter\u00adaction energy with 2O}2 synthon as well as the terminal inter\u00adactions between 4LN2 and BA mol\u00adecules, through hy\u00addroxy-O\u2014H4\u22efN(pyrid\u00adyl) hydrogen bonds, lead to a zigzag electrostatic energy framework, Fig.\u00a06a). The packing system is further stabilized by the dispersion forces contributed by the ten-membered {\u22efHNC2O}2 synthon complemented by other peripheral inter\u00adactions such the pairwise C20\u2014H20\u22efC8/C12\u22efC21, C8\u22efC26 and C6\u2014H6B\u22efO1 inter\u00adactions, which result in a dispersion energy framework resembling a spider web, Fig.\u00a06b). The combination of the electrostatic and dispersion forces leads to an overall energy framework that resembles a ladder, Fig.\u00a06c).Overall, the crystal of (I)Chemical context, 4LH2 mol\u00adecules have long been known to form co-crystals with carb\u00adoxy\u00adlic acids. A list of 4LH2/carb\u00adoxy\u00adlic acid co-crystals is given in Table\u00a044LH2, the length of the central C\u2014C bond, recognized as being long diazen\u00adyl]benzoic acid, carries a hydroxyl residue and this preferentially forms the hydrogen bonds to the pyridyl-N atoms. This observation is contrary to literature expectation where the carb\u00adoxy\u00adlic acid would be expected to form hydrogen bonds preferentially to pyridyl-N atom in instances where there is a competition with putative hydroxyl-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonds loop, in accord with Etter\u2019s rules, i.e. \u2018six-membered ring intra\u00admolecular hydrogen bonds form in preference to inter\u00admolecular hydrogen bonds\u2019 oxalamide (4LH2), was prepared according to the literature; M.p.: 486.3\u2013487.6\u2005K; lit. 486\u2013487\u2005K was mixed with benzoic acid and the physical mixture was then ground for 15\u2005min in the presence of a few drops of methanol. The procedures were repeated three times. Colourless blocks were obtained through careful layering of toluene (1\u2005ml) on an N,N-di\u00admethyl\u00adformamide (1\u2005ml) solution of the ground mixture. M.p.: 435.4\u2013436\u2005K. IR (cm\u22121): 3321 \u03bd(N\u2014H), 3070\u20132999 \u03bd(C\u2014H), 1702\u20131662 \u03bd(C=O), 1506 \u03bd(C=C), 1417 \u03bd(C\u2014N).The precursor, 4LH2:benzoic acid in molar ratios of 1:2, 1:3 and 1:4 were also attempted but only the 2:1 co-crystal (I)Similar experiments with Uiso(H) set to 1.2Ueq(C). The oxygen- and nitro\u00adgen-bound H atoms were located from a difference Fourier map and refined with O\u2014H = 0.84\u00b10.01\u2005\u00c5 and N\u2014H = 0.88\u00b10.01\u2005\u00c5, respectively, and with Uiso(H) set to 1.5Ueq(O) or 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989019016840/hb7875sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019016840/hb7875Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019016840/hb7875Isup3.cmlSupporting information file. DOI: 1972449CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-chromen-4-yl)\u00adoxy]acetic acid di\u00admethyl\u00adsulfoxide monosolvate is described and the inter\u00admolecular contacts in the crystal analysed using Hirshfeld surface analysis and two-dimensional fingerprint plots.The crystal structure of 2-[(2-oxo-2 11H8O5\u00b7(CH3)2SO, is a new coumarin derivative. The asymmetric unit contains two coumarin mol\u00adecules (A and B) and two di\u00admethyl\u00adsulfoxide solvent mol\u00adecules (A and B). The dihedral angle between the pyran and benzene rings in the chromene moiety is 3.56\u2005(2)\u00b0 for mol\u00adecule A and 1.83\u2005(2)\u00b0 for mol\u00adecule B. In mol\u00adecule A, the dimethyl sulfoxide sulfur atom is disordered over two positions with a refined occupancy ratio of 0.782\u2005(5):0.218\u2005(5). In the crystal, mol\u00adecules are linked by O\u2014H\u22efO hydrogen bonds, forming chains running along the c-axis direction. The chains are linked by C\u2014H\u22efO hydrogen bonds, forming layers parallel to the ac plane. In addition, there are also C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions present within the layers. The inter\u00admolecular contacts in the crystal have been analysed using Hirshfeld surface analysis and two-dimensional fingerprint plots, which indicate that the most important contributions to the packing are from H\u22efH (33.9%) and O\u22efH/H\u22efO (41.2%) contacts.The title compound, C The synthesis, and pharmacological and other properties of coumarin derivatives have been studied intensively and reviewed and atoms O2A and O2B deviate from this mean plane by 0.007\u2005(3) and 0.039\u2005(3)\u2005\u00c5, respectively. The dihedral angle between the pyran and benzene rings in the chromene moiety is 3.56\u2005(16)\u00b0 for mol\u00adecule A and 1.83\u2005(16)\u00b0 for mol\u00adecule B; this value is in agreement with those found in analogous coumarin derivatives and 0.218\u2005(5).The 1H-chromen-4-yl)\u00adoxy]acet\u00adyl}piperazin-1-yl)acetamide -2-[(2-oxo-2H-chromen-4-yl)\u00adoxy]acetamide , Cg1\u22efCg4iv = 3.509\u2005(2) and Cg2\u22efCg3iv 3.572\u2005(2)\u2005\u00c5 where Cg1, Cg2, Cg3 and Cg4 are the centroids of rings O1A/C1A/C6A\u2013C9A, C1A\u2013C6A, O1B/C1B/C6B\u2013C9B, and C1B\u2013C6B, respectively; symmetry code: (iv) x, 1\u00a0+\u00a0y, z].The crystal structure features O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds Table\u00a01. In the \u03c0 Table\u00a01 and \u03c0\u2013\u03c0 et al., 2019et al., 2007CrystalExplorer17 through white to blue . The dnorm surface was mapped over a fixed colour scale of 0.774 (red) to 1.381 (blue) for the title compound, where the red spots indicate the inter\u00admolecular contacts involved in the hydrogen bonding.The Hirshfeld surface of the title compound mapped over b) and O\u22efH/H\u22efO at 41.2% , followed by the C\u22efH/H\u22efC contacts at 9.6% , C\u22efC contacts at 6.3% and S\u22efH/H\u22efS contacts at 3.9% .The fingerprint plots are given in Fig.\u00a062% Fig.\u00a06c, follo6% Fig.\u00a06d, C\u22efC c3% Fig.\u00a06e and S\u22ef9% Fig.\u00a06f.et al., 2016et al., 2006H-furo-[1]-benzo\u00adpyran-7-one monohydrate -4,5-bis\u00ad(pyrrolidinylcarbon\u00adyl)-2,2-dimethyl-1,3-dioxolane cyclo\u00adpropyl sulfamate acetate in THF (10\u2005mL) at 273\u2005K and stirred at 273\u2005K for 1\u2005h. Completion of the reaction was confirmed by TLC (mobile phase ethyl acetate/hexa\u00adne) and THF was distilled off using a rotavapor. The obtained solution was washed with ethyl acetate (20\u2005mL). The aqueous layer was acidified with 2N HCl (pH 1.0\u20132.0) and the obtained solid was filtered, washed with hexane and dried under vacuum to give as white solid. The purified compound was recrystallized using dimethyl sulfoxide as solvent.A solution of lithium hydroxide in water (4\u2005mL) was added to ethyl 2-(2-oxo-2Uiso(H) = 1.5Ueq(C-meth\u00adyl) or 1.2Ueq(C) for other H atoms. In mol\u00adecule A, the sulfur atom of the sulfinyldi\u00admethane group is disordered over two positions with refined occupancies of 0.782\u2005(5) and 0.218\u2005(5). In the final cycles of refinement, five outliers were omitted.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019009447/vm2218sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019009447/vm2218Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019009447/vm2218Isup3.cmlSupporting information file. DOI: 1891495CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, N\u2014H\u22efCl, O\u2013H\u22efCl and N\u2014H\u22efO hydrogen bonds connect the anions, cations and water mol\u00adecules, forming a three-dimensional network.In this hydrated 2,6,13,17-tetra\u00adazoniatri\u00adcyclo\u00ad[16.4.0.0 20H44N44+\u00b74Cl\u2212\u00b74H2O, has been determined using synchrotron radiation at 220\u2005K. The structure determination reveals that protonation has occurred at all four amine N atoms. The asymmetric unit contains one half-cation , two chloride anions and two water mol\u00adecules. There are two mol\u00adecules in the unit cell. The Cl\u2212 anions and hydrate mol\u00adecules are involved in hydrogen bonding. The crystal structure is stabilized by inter\u00admolecular hydrogen bonds involving the macrocycle N\u2014H groups and water O\u2014H groups as donors and the O atoms of the water mol\u00adecules and the Cl\u2212 anions as acceptors, giving rise to a three-dimensional network.The crystal structure of the title salt, C These di- or tetra\u00adammonium cations may be suitable for the removal of toxic heavy metal ions from water. The macrocycle L contains a cyclam backbone with two cyclo\u00adhexa\u00adne subunits. Methyl groups are attached to the 3 and 14 carbon atoms of the propyl chains that bridge opposite pairs of N atoms in the structure. Previously, we have reported the crystal structures of [Cu(L)](NO3)2\u00b73H2O, [Cu(L)](NO3)2, [Cu(L)](ClO4)2 and [Cu(L)(H2O)2](BF4)2\u00b72H2O together with [Zn(L)(OCOCH3)2]. In these structures, the copper(II) or zinc(II) cations have tetra\u00adgonally distorted octa\u00adhedral environments with the four N atoms of the macrocyclic ligand in equatorial positions and O atoms of counter-anions, water mol\u00adecules or acetato ligands in axial positions \u00b72MeOH The macrocycle 3,14-dimethyl-2,6,13,17-tetra\u00adaza\u00adtri\u00adcyclo\u00ad(16.4.0.0\u2212 anions and four solvent water mol\u00adecules and was characterized during studies of the macrocyclic ligand and its copper(II) complexes. An ellipsoid plot of the mol\u00adecular components in (I)anti with respect to the macrocyclic plane as a result of the mol\u00adecular inversion symmetry. The six-membered cyclo\u00adhexane ring is in a stable chair conformation. Within the centrosymmetric tetra-protonated amine unit [C20H44N4]4+, the C\u2014C and N\u2014C bond lengths vary from 1.522\u2005(2) to 1.542\u2005(2)\u2005\u00c5 and from 1.506\u2005(2) to 1.522\u2005(2)\u2005\u00c5, respectively. The ranges of N\u2014C\u2014C and C\u2014N\u2014C angles are 106.85\u2005(10) to 114.32\u2005(11)\u00b0 and 116.70\u2005(10) to 118.89\u2005(10)\u00b0, respectively. The bond lengths and angles within the [C20H44N4]4+ tetra-cation are comparable to those found in the free ligand or the di-cation in C20H40N4\u00b72C11H10O \u00b72MeOH 3}(CN)3]2\u00b72H2O\u00b72MeOH . The crystal structures of C20H40N4\u00b72C11H10O \u00b72MeOH 3}(CN)3]2\u00b72H2O\u00b72MeOH were used as provided. All chemicals were reagent grade and used without further purification. As a starting material, the macrocycle 3,14-dimethyl-2,6,13,17-tetra\u00adaza\u00adtri\u00adcyclo\u00addocosane was prepared according to a published procedure in water (15\u2005mL). The solution was heated for 1\u2005h at 338\u2005K. After cooling to 298\u2005K, the pH was adjusted to 3.0 with 1.0 M HCl. The solution was filtered and left at room temperature. Colourless crystals suitable for X-ray analysis were obtained unexpectedly from the solution over a period of a few days.Commercially available Uiso(H) values of 1.2Ueq and 1.5Ueq(C-meth\u00adyl). O-bound H atoms of the water mol\u00adecules were located in a difference-Fourier map, and the O\u2014H distances and the H\u2014O\u2014H angles were restrained using DFIX and DANG constraints (0.84 and 1.36\u2005\u00c5).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018009337/sj5559sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018009337/sj5559Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018009337/sj5559Isup3.cmlSupporting information file. DOI: 1852146CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Cav1.2 is the pore\u2010forming subunit of L\u2010type voltage\u2010gated calcium channel (LTCC) that plays an important role in calcium overload and cell death in Alzheimer's disease. LTCC activity can be regulated by estrogen, a sex steroid hormone that is neuroprotective. Here, we investigated the potential mechanisms in estrogen\u2010mediated regulation of Cav1.2 protein. We found that in cultured primary neurons, 17\u03b2\u2010estradiol (E2) reduced Cav1.2 protein through estrogen receptor \u03b1 (ER\u03b1). This effect was offset by a proteasomal inhibitor MG132, indicating that ubiquitin\u2013proteasome system was involved. Consistently, the ubiquitin (UB) mutant at lysine 29 (K29R) or the K29\u2010deubiquitinating enzyme TRAF\u2010binding protein domain (TRABID) attenuated the effect of ER\u03b1 on Cav1.2. We further identified that the E3 ligase Mdm2 (double minute 2 protein) and the PEST sequence in Cav1.2 protein played a role, as Mdm2 overexpression and the membrane\u2010permeable PEST peptides prevented ER\u03b1\u2010mediated Cav1.2 reduction, and Mdm2 overexpression led to the reduced Cav1.2 protein and the increased colocalization of Cav1.2 with ubiquitin in cortical neurons in vivo. In ovariectomized (OVX) APP/PS1 mice, administration of ER\u03b1 agonist PPT reduced cerebral Cav1.2 protein, increased Cav1.2 ubiquitination, and improved cognitive performances. Taken together, ER\u03b1\u2010induced Cav1.2 degradation involved K29\u2010linked UB chains and the E3 ligase Mdm2, which might play a role in cognitive improvement in OVX APP/PS1 mice. This effect involved lysine 29\u2010linked ubiquitin chains and the E3 ligase Mdm2 (double minute 2 protein). In OVX APP/PS1 mice, systematic administration of E2 and ER\u03b1 agonist PPT (propylpyrazoletriol) led to the reduced Cav1.2 protein and the enhanced Cav1.2 ubiquitination in the brain, which were accompanied by the improved cognitive functions.22.1Ca2+density) were measured using whole\u2010cell patch recordings. As shown in Figure Ca2+density (pA/pF) was significantly decreased by E2 treatment (24\u00a0hr), whereas ICI attenuated this effect. These results indicated that E2 reduced Cav1.2 protein in cultured primary cortical neurons, which was mediated by ERs.As shown in Figure Ca2+ density might mediate E2 effect on Cav1.2, we tested the effect of the selective ER\u03b1 agonist or ER\u03b2 agonist on Cav1.2 incubated in cortical neurons for 24\u00a0hr. As shown in Figure y Figure . As expe2.2Cav1.2 protein can be regulated at multiple levels controls Cav1.2 degradation protein contains seven lysine (K) residues at positions 6, 11, 27, 29, 33, 48, and 63, respectively. In addition, poly\u2010UB chain assembly can occur at any of these lysine residues and CHIP are reportedly associated with ER\u03b1 signaling mice were significantly shorter than that in saline\u2010treated OVX APP/PS1 (AD) mice beginning on the third day Figure a\u2013c. In t3The efficacy of estrogen\u2010based hormone therapy (HT) in AD is not conclusive. Some studies reveal that HT may reduce the risk of AD in postmenopausal women mediates proteasomal degradation of Cav1.2 in neuronal cells were from Abcam and Alomone, respectively, which produced identical Western blots. Those against \u03b2\u2010actin (ab6276), MAP\u20102 (ab32454), ER\u03b1 (ab3573), ER\u03b2 (ab104804), calcineurin (ab3673), Mdm2 (ab16895), and ubiquitin (ab19247) were purchased from Abcam. pSer1928\u2010Cav1.2 (A010\u201070) was from Badrilla (United Kingdom). 17\u03b2\u2010estradiol (E2758), propylpyrazoletriol , diarylprepionitrile , E2\u2010BSA (E5630), ICI 182,780 (V900926), nifedipine (N7630), FK506 (F4679), MG132 (C2211), ammonium chloride (A0171), chloroquine (C6628), carfilzomib (791938), and other reagents were purchased from Sigma . Lipid solvents were made in stock solutions in DMSO .4.2pCMV6\u2010Myc\u2010Mdm2 and pCMV6\u2010Myc\u2010p14ARF were purchased from OriGene (USA). TRABID (DU# 49067) and TRABID mutant C443A (DU# 49089) were purchased from MRC PPU Reagents and Services (University of Dundee). pRK5\u2010HA\u2010ubiquitin was obtained from Addgene. All ubiquitin mutants were made from pRK5\u2010HA\u2010ubiquitin plasmid by mutagenesis. pRK5\u2010HA\u2010Ubiquitin\u2010K0 (no lysine), pRK5\u2010HA\u2010Ubiquitin\u2010K6 (keep lysine at position 6), pRK5\u2010HA\u2010Ubiquitin\u2010K11, pRK5\u2010HA\u2010Ubiquitin\u2010K27, pRK5\u2010HA\u2010Ubiquitin\u2010K29, pRK5\u2010HA\u2010Ubiquitin\u2010K33, pRK5\u2010HA\u2010Ubiquitin\u2010K48, pRK5\u2010HA\u2010Ubiquitin\u2010K63, pRK5\u2010HA\u2010Ubiquitin\u2010K6R , pRK5\u2010HA\u2010Ubiquitin\u2010K29R, pRK5\u2010HA\u2010Ubiquitin\u2010K63R, CHIP with myc tag, and Derlin\u20101 with HA tag were cloned to pcDNA3, respectively.4.36\u00a0cells/ml for biochemical experiments and 0.5\u00a0\u00d7\u00a0106\u00a0cells/ml for molecular and immunohistochemistry experiments. Cultures were maintained in serum\u2010free neurobasal (NB) medium at 37\u00b0C with 5% CO2. Fifty percent of the medium was exchanged with fresh medium every other day for 2\u00a0weeks. On the day before experiment, culture medium was completely replaced by fresh medium in the presence of accurate dilution of chemicals.Cortical neurons were extracted from prenatal pups on embryonic day 18. Cells were placed on poly\u2010L\u2010lysine (0.1\u00a0mg/ml)\u2010coated 6\u2010well plates, at a density of 2\u00a0\u00d7\u00a0104.42. At\u00a0~\u00a090% confluence, cells were transiently transfected with various plasmids or Mdm2 siRNA using the Lipofectamine 2000 reagent (Invitrogen) according to the manufacturer's instructions. A total of 2.5\u00a0\u03bcg of DNA in 100\u00a0\u03bcl Opti\u2010MEM (Invitrogen) were used in each of six wells. The following human Mdm2 siRNA sequences were used for transfection in SH\u2010SY5Y cells: sense CGUACGCGGAAUACUUCGATT, antisense AAUCGAAGUAUUCCGCGUACG; NC: sense CGUACGCGGAAUACUUCGATT, antisense AAUCGAAGUAUUCCGCGUACG (100\u00a0pM per six wells for 72\u00a0hr). The effectiveness of Mdm2 knockdown was verified by Western blotting analysis.HT22 and SH\u2010SY5Y (human neuroblastoma) cell lines were obtained from the Type Culture Collection of the Chinese Academy of Sciences . Cells were incubated in Dulbecco's modified Eagle's medium containing 4.5\u00a0g glucose/L, supplemented with penicillin/streptomycin (50\u00a0units/mL), glutamate (2\u00a0mM), and 10% fetal bovine serum , and were maintained at 37\u00b0C with 10% CO4.5Western blotting of Cav1.2 was performed as previously described assays, the cell or tissue extracts (0.5\u00a0mg) were pre\u2010incubated with protein A agarose beads for 3\u00a0hr to reduce nonspecific reaction. The supernatants were then mixed with nonspecific IgG and appropriate antibodies for 1\u00a0hr. The mixtures were then incubated with protein A agarose beads overnight at 4\u00b0C. Samples were washed two times with lysis buffer and two times with sterile PBS and denatured with SDS sampling buffer.4.6Brain sections were washed with ice\u2010cold PBS, then fixed in ice\u2010cold 4% paraformaldehyde for 10\u00a0min, permeabilized in 0.1% Triton X\u2010100 for 10\u00a0min, and blocked with 10% normal goat serum for 1\u00a0hr at 37\u00b0C with extensive PBS washings between each step. Specimens were incubated with appropriate primary antibodies (1:50 in PBS) at 4\u00b0C overnight. The sections were then washed 6\u00a0\u00d7\u00a02\u00a0min with PBS, followed by incubation with the secondary antibody Alexa 549 or Fluro 488 (1:100 in PBS) at 37\u00b0C for 1\u00a0hr. After sealing with a mixture of glycerol and PBS at a ratio of 1:1, the immunofluorescent\u2010labeled sections were examined using a laser scanning confocal microscope with an Olympus IX 70 inverted microscope equipped with a Fluoview FVX confocal scan head.4.72+) were recorded from cultured cortical neurons from 7 to 11\u00a0days in vitro (DIV), at room temperature using a MultiClamp 700B amplifier (Axon Instruments). The recording chamber was perfused with Tyrode's solution containing (in mM): NaCl 100, tetraethylammonium\u2010Cl 20, 4\u2010AP 5, KCl 4, MgCl2 1, BaCl210, glycine 0.01, HEPES 25, glucose 30, and TTX 0.001. The glass micropipettes had a resistance of 3\u20135\u00a0M\u03a9 when filled with internal solution containing (in mM): CsCl 135, MgCl2 4, HEPES 10, EGTA 10, MgATP 4, and Na3GTP 0.3. A prepulse protocol consists of 300\u2013700\u00a0ms to \u221230\u00a0mV followed by 50\u00a0ms to \u221250\u00a0mV, before each test pulse was used to inactivate T\u2010type Ca2+ channels and Na+ channels. ICa2+ was elicited by 300\u00a0ms test pulses of variable amplitude (\u221240 to +60\u00a0mV at a step of 10\u00a0mV) from a holding potential of \u221260\u00a0mV. The interval between test pulses was 10\u00a0s.Voltage\u2010dependent calcium\u2010mediated currents . The primer sequences for Cav1.2 were as follows: 5\u2032\u2010TACCGTCAGTTCCACACAGC\u20103\u2032 and 5\u2032\u2010CTTCAGAGTCAGGCAGAGCA\u20103\u2032. The threshold cycle (Ct) value of each sample was calculated, and the relative mRNA level was normalized to the GAPDH mRNA value.4.9ER\u03b1/\u03b2 shRNAs was produced by Shanghai GeneChem Co., Ltd. . At 10 DIV, ER\u2010related lentivirus was added to the culture for 72\u00a0hr according to the manufacturers\u2019 instruction , while transfection of Mdm2 was performed in HT22 cells.The target shRNAs against rat ER\u03b1/\u03b2 gene (NM_012689/NM_012754) were designed as follows: ER\u03b1\u20101: GATAAGAACCGGAGGAAGA, ER\u03b1\u20102: CCAGAATGGCCGAGAGAGA, and ER\u03b1\u20103: GTAAATGTGTAGAAGGCAT; ER\u03b2\u20101: TGAGCAAAGCCAAGAGAAA, ER\u03b2\u20102: TCACTAAGCTGGCGGACAA, and ER\u03b2\u20103: CCAAATGTGCTATGGCCAA. Negative control was the scrambled sequence with no homology to rat genes. The recombinant lentiviral vector bearing rat 4.10in utero electroporation was performed on pregnant C57/B6L mice on embryonic day 14.5 (E14.5) as previously described and PEST3 (S840/861) were synthesized and purified by Pepmic Co. Ltd . The sequences were as follows: YGRKKRRQRRR\u2010DIDPENEDEGMDED\u2010GFP for TAT\u2010PEST1, YGRKKRRQRRR\u2010DIDKENEDEGKDED\u2010GFP (\u0394PEST1) for scrambled, YGRKKRRQRRR\u2010HSNPNTAGEEDEEEPEMPVGPR\u2010GFP for TAT\u2010PEST3, and YGRKKRRQRRR\u2010HSNPNPAGEEDKEEPAMPVGPR\u2010GFP (\u0394PEST3) for scrabbled control were obtained from Nanjing University (China). Eight\u2010month\u2010old female APP/PS1 mice were bilaterally ovariectomized (OVX) for 14\u00a0days. Animals were subcutaneously injected daily with vehicle, 17\u03b2\u2010estradiol , PPT (1\u00a0mg/kg), and DPN (1\u00a0mg/kg) for two weeks. The dosage of E2 is thought to be within physiological range; and those of PPT and DPN are functional were recorded by a video and analyzed by image analyzing software . For contextual and cued fear conditioning, mice were tested in a 3\u2010day paradigm. Behavior was recorded by video camera, and freezing data were measured using FreezeScan software.4.14SEM from at least three independent experiments. The statistical comparisons between two groups were tested using Student's t test. The comparisons among groups were tested using one\u2010way or two\u2010way ANOVA, and a post hoc pairwise comparison was used where it applied.Data were presented as means\u00a0\u00b1\u00a0None declared.G\u2010J Chen and Z Yan designed the research; Y\u2010J Lai performed the research and analyzed the data; B\u2010L Zhu, F Sun, D Luo, Y\u2010L Ma, B\u2010Luo, J Tang, M\u2010J Xiong, L Liu, X\u2010T Hu, L He, X\u2010J Deng, J\u2010H Zhang, and J Yang provided assistance with the research; G\u2010J Chen and Y\u2010J Lai wrote the paper. All authors read and approved the final manuscript.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file."} +{"text": "R)\u2010[11C]PK11195 was investigated. The synthesis of the reference compound was accomplished in six steps and 9\u2009% overall yield, and the precursor was prepared in eight steps and 8\u2009% overall yield. The chiral separation of the reference and precursor compounds was performed using supercritical fluid chromatography with >95\u2009%\u2005ee. The absolute configuration was determined by circular dichroism. Optimisation of reaction conditions for manual radiolabelling revealed acetonitrile as a preferred solvent at 100\u2009\u00b0C. Automation of this radiolabelling method provided R and S enantiomers in respective 21.3\u00b116.7 and 25.6\u00b17.1\u2009% decay\u2010corrected yields and molar activities of 55.8\u00b135.6 and 63.5\u00b139.5\u2005GBq\u2009\u03bcmol\u22121 (n=3). Injection of the racemic analogue into a healthy rat confirmed passage through the blood\u2013brain barrier.Translocator protein (TSPO) is a biomarker of neuroinflammation, which is a hallmark of many neurodegenerative diseases and has been exploited as a positron emission tomography (PET) target. Carbon\u201011\u2010labelled PK11195 remains the most applied agent for imaging TSPO, despite its short\u2010lived isotope and low brain permeability. Second\u2010generation radiotracers show variance in affinity amongst subjects caused by the genetic polymorphism (rs6971) of the TSPO gene. To overcome these limitations, a new structural scaffold was explored based on the TSPO pharmacophore, and the analogue with a low\u2010affinity binder/high\u2010affinity binder (LAB/HAB) ratio similar (1.2 vs. 1.3) to that of ( R\u20101\u2010(2\u2010chlorophenyl)\u2010N\u2010methyl\u2010N\u2010(1\u2010methylpropyl)\u20103\u2010isoquinoline carboxamide ((R)\u2010[11C]PK11195, [11C]1, Figure\u2005m).R)\u2010[11C]1 has been used in PET imaging to show neuroinflammation in\u2005vivo in cases of stroke, neurodegeneration, traumatic brain injury, and neoplasia.11C]1 in research and clinical settings, such as low brain permeability,t1/2=20\u2005min), which restricts its use to PET centres that have a cyclotron on site.Neuroinflammation, an immune response to neuronal insult, is a core component of many disorders such as stroke, multiple sclerosis (MS), neurodegenerative disorders, and brain tumours.11C]1, multiple new TSPO radiotracers have been developed and studied; these have improved properties for application, including several that are labelled with the longer\u2010lived radioisotope fluorine\u201018 (t1/2=109.9\u2005min).N\u2010(2\u2010(methoxy\u201011C)benzyl)\u2010N\u2010(4\u2010phenoxypyridin\u20103\u2010yl)acetamide , and good clearance from striatum (region of low TSPO expression) in rats.In 2004, a study by Okubo et\u2005al. described a tetracyclic indole\u2010based pharmacophore .However, while 6 as well as the initial in\u2005vivo preclinical PET.Toward this objective informed by structure\u2013activity design features, a novel series of compounds based on the chemical scaffold18F]6, we proposed an SN2 nucleophilic substitution of a good leaving group with [18F]fluoride, for which reason tosylate 25 and mesylate 26 were envisioned as potential precursors. Reference compound 6 to give product 23. A second hydrogenation on crude 23 in the absence of triethylamine was performed to remove the benzyl protecting group to give crude 24 in quantitative yield. Using crude alcohol 24, we prepared both crude tosylate 25 and crude mesylate precursor 26 . However, the mesyl precursor 26 required purification by column chromatography, yielding a viscous oil (37\u2009%), thus making tosyl precursor 25 practically easier to purify and handle than the mesyl precursor 26.Analogously, 2\u2010chloro\u20105\u2010methoxyaniline hydrochloride 18F]6 by establishing the chiral separation of both reference compound 6 as well as its radiolabelling precursor (25). A clear choice for chiral separation was found in the preparation of diastereomeric mixture by application of a chiral derivatisation group. We identified an acid intermediate 21 for further functionalisation with several potential enantiomerically pure alcohols. The alcohols selected were l\u2010menthol, (R)\u2010(\u2212)\u20102\u2010butanol, and (R)\u20101\u2010phenyl\u20102\u2010propyn\u20101\u2010ol. The ester 28 with (R)\u2010(\u2212)\u20102\u2010butanol was formed in 53\u2009% yield using DBU as a base and a mesylate analogue 27 of the alcohol as an intermediate. However, a higher\u2010yielding and a more direct method of EDCI/DMAP coupling was used to form the esters with l\u2010menthol and (R)\u20101\u2010phenyl\u20102\u2010propyn\u20101\u2010ol, which afforded 29 and 30 in 54 and 31\u2009% yields, respectively \u20101\u2010phenyl\u20102\u2010propynyl analogues. Flash chromatography using diethyl ether and cyclohexane yielded two diasteriomerically enriched samples with both esters 29 (in 78 and 40\u2009% diastereomeric excess) and 30 (in 82 and 35\u2009% diastereomeric excess) \u201029, \u201029 and \u201030, \u201030). To demonstrate the presence of a single enantiomer in a mixture, the chiral l\u2010menthol functionality was first removed under a variety of basic conditions (Table\u20052O in EtOH (entry\u20051), which were the hydrolytic conditions used in the synthesis of the reference compound 6 and precursors 25 and 26. Reaction with LiOH in a mixture of THF/H2O (entry\u20052) gave a minor byproduct.While NMR analysis demonstrated the presence of two diastereomers in all three product mixtures, TLC analysis of the ester mixtures using a variety of solvent systems showed the two diastereomers could be separated readily by flash column chromatography in two cases, that of 2CO3 in MeOH (entry\u20053) gave an equal amount of the desired product and an unidentified byproduct,l\u2010menthol using EDCI/DMAP. To our disappointment, 1H\u2005NMR analysis indicated complete racemisation from all investigated hydrolytic reaction conditions. This was further confirmed by chiral HPLC analysis of the acid samples obtained from hydrolysis of diastereomerically enriched ester samples \u201021, Scheme\u200521 in a solution presumably due to the presence of a highly acidic proton is a likely explanation for such rapid racemisation of the stereocenter. In the case of the ester analogue 30, while we were able to obtain diastereomerically enriched samples through multiple rounds of flash column chromatography in diastereomeric excess \u201030, \u201030), a previously described CuCl hydrolysis protocol failed to give the enantiomerically enriched product 21 (Scheme\u20056 and tosylate 25 (see the Supporting Information for details).Using K6 was next determined by electronic circular dichroism (CD) spectroscopy whereby CD spectra of 6 were compared with that of 5, for which stereochemistry had already been established \u20105 was composed of a maximum positive Cotton effect at 200\u2005nm and a corresponding maximum negative Cotton effect at 226\u2005nm. The enantiomer of 6, which similarly exhibited a maximum positive Cotton effect at 204\u2005nm and a corresponding maximum negative Cotton effect at 228\u2005nm, was in analogy assigned as the S\u2005isomer. The other isomer showed the mirror image bisignate band in the CD spectrum and was therefore identified as R\u2005enantiomer.The absolute stereochemistry of enantiomers of d Figure\u2005. The speS\u2005enantiomer (Supporting Information).To correlate enantiomers obtained via SFC separation to CD data, the chiral HPLC separation was performed on a Whelk\u2010O1 column to unambiguously identify first eluting peak as the 5 was shown to bind to wild\u2010type TSPO with a higher affinity than compound 1, but lost affinity at A147T TSPO (Table\u2005S)\u20106 bound to wild\u2010type TSPO with a similar affinity to compound 1, and both compounds 1 and (S)\u20106 only showed minor loss in affinity at A147T TSPO \u20106: 36.3\u2005nm, while 1.04\u2005nm for rat TSPO.Using an assay based on human embryonic kidney cell lines stably overexpressing human TSPO wild\u2010type and TSPO A147TPO Table\u2005. Compoun18F]5, manual radiochemical reactions were performed first. Both precursors 25 and 26 were assessed and the effect of temperature, solvent, and time was investigated (Table\u200525 (entries\u20051\u20136), a single radiolabelled product formed which was identified as [18F]6 via co\u2010injection of reference compound 6. However, when using 26 (entries\u20057\u201312), a second unidentified radiolabelled product was formed during the reaction. The amount of this radiolabelled side product which formed immediately, did not change during the course of the reaction.To establish suitable reaction conditions for radiolabelling of [ed Table\u2005. For all18F]fluoride to [18F]6 was observed: the highest being 18\u2009% when MeCN was used as a solvent and mesylate 25 as a precursor. When the temperature was increased to 100\u2009\u00b0C or 120\u2009\u00b0C reactions performed in DMF similarly showed poor conversion (entries\u20056 and 12) after 20\u2005min. In DMSO, [18F]6 formed with 3\u2009% conversion after 20\u2005min (entry\u200510) when 26 was used as precursor, whereas tosylate 25 afforded 47\u2009% conversion into [18F]6 under analogous conditions (entry\u20054). The highest conversion in the case of both precursors 25 and 26 was observed after 20\u2005min in MeCN: 81 and 39\u2009%, respectively (entries\u20052 and 8).At 80\u2009\u00b0C, independent of solvent and precursor, low conversion of <20\u2009% of [25 was used for automated radiolabelling on a GE Healthcare FXFN TRACERlab instrument. [18F]Fluorination was achieved with cyclotron\u2010produced [18F]fluoride in MeCN at 100\u2009\u00b0C for 20\u2005min. Product [18F]6 was purified by semipreparative reversed\u2010phase HPLC.With this result in hand , next tosylate 18F]6 was 60\u2005min from end of bombardment (EOB). Radiochemical purity was over 99\u2009%, and radiolabelling yield was 20.8\u00b14.5\u2009% decay\u2010corrected to EOB or 13.6\u00b12.8\u2009% non\u2010decay\u2010corrected, and molar radioactivity 93.2\u00b150.6 GBq\u2009\u03bcmol\u22121 (n=9) at the end of synthesis (EOS). Furthermore, the same automated protocol was used for the radiosyntheses of (R)\u2010[18F]6 and (S)\u2010[18F]6 as follows: 21.3\u00b116.7 and 25.6\u00b17.1\u2009% decay\u2010corrected yields, radiochemical purity >98\u2009% and molar radioactivities 55.8\u00b135.6 and 63.5\u00b139.5\u2005GBq\u2009\u03bcmol\u22121 (n=3) at the EOS, respectively.Total radiosynthesis time for the preparation for [18F]6 enters the brain was distilled over P2O5 and stored over KOH. Reported density values are for ambient temperature. Purity of compounds was \u226595\u2009% as determined by analytical HPLC on a Thermo Dionex 3000 HPLC system. Preparative chromatographic separations were performed on Material Harvest silica gel 60 (35\u201375\u2005\u03bcm) and reactions followed by TLC analysis using Sigma\u2013Aldrich silica gel 60 plates (2\u201325\u2005\u03bcm) with fluorescent indicator (254\u2005nm) and visualised with UV or potassium permanganate.All reactions requiring anhydrous conditions were conducted in oven\u2010dried glass apparatus under an atmosphere of inert gas. Reagents were purchased from Sigma\u2013Aldrich or Alfa Aesar. Reagents were used without further purification unless otherwise noted. Triethylamine =7.26\u2005ppm, \u03b4C=77.2\u2005ppm. Multiplicities in the 1H\u2005NMR spectra are described as: s=singlet, d=doublet, t=triplet, q=quartet, quint.=quintet, s=sextet, h=heptet, m=multiplet, b=broad; coupling constants are reported in Hz.18 1.7\u2005\u03bcm, 2.1\u00d750\u2005mm; flow rate: 0.6\u2005mL\u2009min\u22121, 4\u2005min method, 5\u201395\u2009% MeCN/H2O with 0.1\u2009% formic acid. HRMS data were obtained from the EPSRC Mass Spectrometry Service at the University of Swansea. Ion mass/charge (m/z) ratios are reported as values in atomic mass units.LC\u2013MS data were obtained using a Waters Acquity\u2010H/Xevo TQD LC\u2013MS instrument. Column: Acquity UPLC\u00ae BEH CFN TRACERlab. Semipreparative purification of radiolabelled material was performed on a Merck\u2010Hitachi L6200A system equipped with Knauer variable wavelength detector and an Eberline radiation detector using a reversed\u2010phase column and eluting with 48\u2009% aq. MeCN at a flow rate of 5\u2005mL\u2009min\u22121. Analytical HPLC samples were analysed by an Agilent HPLC 1100 system equipped with UV multi\u2010wavelength detector and Raytest Gabi star radiation detector using reversed\u2010phase column (Phenomenex Luna 5\u2005\u03bcm C18 (2) 100\u2005\u00c5, 250\u00d74.6\u2005mm, 675295\u201011) and eluting with 0\u20133\u2005min isocratic 10\u2009% aq. MeCN, 3\u20135\u2005min gradient 10\u201380\u2009% aq. MeCN, 6\u201312\u2005min isocratic 80\u2009% aq. MeCN at a flow rate of 1.5\u2005mL\u2009min\u22121.The automated radiosynthesis was performed on a GE Healthcare FX2\u2010Fluoroethyl 4\u2010methylbenzenesulfonate (8). Commercially available 2\u2010fluoroethanol 7 was dissolved in pyridine (2.5\u2005mL) under nitrogen. The solution was stirred at 0\u2009\u00b0C and tosyl chloride added portion\u2010wise to the solution over a period of 30\u2005min, keeping the temperature below 5\u2009\u00b0C. The reaction was stirred at RT for 18\u2005h. The reaction was then quenched by careful addition of ice followed by water (5\u2005mL). The reaction mixture was extracted into EtOAc (10\u2005mL) and washed with water (3\u00d710\u2005mL), 1\u2009m HCl solution (10\u2005mL), 1\u2009m aqueous sodium carbonate (10\u2005mL), and copper sulfate (2\u00d710\u2005mL). The organic layer was washed with brine (10\u2005mL), dried (MgSO4) and concentrated in\u2005vacuo to give 8 as an oil . LC\u2013MS: Rf 1.86 (\u2212ESI) m/z 311.5 ([M+94]+); 1H\u2005NMR : \u03b4H=7.81 , 7.36 , 4.57 , 4.26 , and 2.45 . Characterization data for this compound are in complete agreement with previously published data.N\u2010(2\u2010fluoroethyl)\u20105\u2010methoxyaniline (10)2\u2010Chloro\u2010. Commercially available 2\u2010chloro\u20105\u2010methoxyaniline hydrochloride 9 was dissolved in DMF (60\u2005mL) and sodium hydride was added. The reaction was stirred for 30\u2005min at RT under nitrogen. The tosylate 8 in DMF was then added dropwise and the reaction was stirred at RT for 2\u2005h. The reaction was then heated at 100\u2009\u00b0C for 18\u2005h. The reaction was allowed to cool and the solvent was removed under reduced pressure. The residue was dissolved in EtOAc (100\u2005mL) and washed with water (5\u00d7100\u2005mL). The organic layers were combined, dried (MgSO4) and concentrated in\u2005vacuo to give a brown oil. The crude mixture was then purified by silica flash chromatography eluting with a gradient of 5\u201330\u2009% EtOAc/petroleum ether (40\u201360\u2009\u00b0C) to yield 10 as a yellow oil . LC\u2013MS: Rf 2.01 (+ESI) m/z 204.3 ([M+H]+); 1H\u2005NMR : \u03b4H=7.16 CHCH), 6.25\u20136.22 CHCH), 4.64 , 4.65\u20134.55 , 3.77 , and 3.48 . Characterization data for this compound are in complete agreement with an internal GE publication.Ethyl 3\u2010bromo\u20102\u2010hydroxycyclohex\u20101\u2010ene\u20101\u2010carboxylate (11). Ethyl 2\u2010oxocyclohexanecarboxylate S1 was dissolved in diethyl ether and cooled to 0\u2009\u00b0C under nitrogen. Bromine was added dropwise over 15\u2005min and the reaction mixture was allowed to warm to RT over 90\u2005min. The mixture was slowly poured into ice\u2010cold saturated aqueous potassium carbonate and extracted with ethyl acetate. The combined organic layers were dried over magnesium sulfate, filtered, concentrated in\u2005vacuo and dried on the vacuum line for 18\u2005h to afford 11 as a yellow oil. LC\u2013MS: Rf 2.17; 1H\u2005NMR : \u03b4H=11.92 , 4.93 ), 4.20 OCH2CH3), 2.53\u20132.46 , 2.12\u20132.03 , 1.79\u20131.67 , 1.24 OCH2CH3). Characterization data for this compound are in complete agreement with an internal GE publication.Ethyl\u20103\u2010((2\u2010chloro\u20105\u2010methoxyphenyl)(2\u2010fluoroethyl)amino)\u20102\u2010hydroxycyclohex\u20101\u2010ene\u20101\u2010carboxylate (12). A solution of the aniline 10 in THF (60\u2005mL) was cooled to \u221240\u2009\u00b0C. Potassium bis(trimethylsilyl)amide was added dropwise and the reaction stirred for 30\u2005min at \u221240\u2009\u00b0C. The carboxylate 11 in THF (10\u2005mL) was added dropwise at \u221240\u2009\u00b0C. The cooling bath was removed and the reaction was stirred at RT for 4\u2005h. The reaction was quenched with brine (100\u2005mL) and extracted into EtOAc (2\u00d7200\u2005mL), dried (MgSO4) and concentrated in\u2005vacuo to give 12 as a brown oil which was used crude in the next step. LC\u2013MS: Rf 2.29 (+ESI) m/z 372.4 ([M+H]+). Characterization data for this compound are in complete agreement with an internal GE publication.H\u2010carbazole\u20104\u2010carboxylate (13)Ethyl 8\u2010chloro\u20109\u2010(2\u2010fluoroethyl)\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101. The intermediate 12 was dissolved in Et2O (100\u2005mL) and zinc chloride was added. The reaction was heated at reflux for 16\u2005h. EtOAc (300\u2005mL) was added to dissolve everything and was washed with 2\u2009m HCl (200\u2005mL), water (200\u2005mL), 10\u2009% aqueous potassium carbonate (200\u2005mL), dried (MgSO4) and concentrated in\u2005vacuo. The crude material was purified by silica gel chromatography eluting with a gradient of 5\u201320\u2009% of EtOAc/petroleum ether (40\u201360\u2009\u00b0C) to afford 13 as a yellow oil . Material could not be obtained in a pure form, but regardless, the reaction was progressed to the next step. LC\u2013MS: Rf 2.38 (+ESI) m/z 354.4 ([M+H]+); 1H\u2005NMR : \u03b4H=6.95 , 6.35 , 4.90\u20134.40 , 4.20\u20134.10 , 2.80\u20132.65 , 3.79 , 2.10\u20131.80 , and 1.30\u20131.20 . Characterization data for this compound are in complete agreement with an internal GE publication.H\u2010carbazole\u20104\u2010carboxylate (14)Ethyl 9\u2010(2\u2010fluoroethyl)\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101. The chloro intermediate 13 was dissolved in methanol (50\u2005mL) and triethylamine and 10\u2009% Pd/C (0.414\u2005g) were added. The mixture was stirred for 18\u2005h after purging twice (over 2\u2005h) with hydrogen gas under atmospheric pressure. The reaction was filtered through a pad of Celite under nitrogen atmosphere, washed with methanol (50\u2005mL), and the solvent was removed in\u2005vacuo. The residue was dissolved in EtOAc (100\u2005mL) and washed with 10\u2009% aqueous potassium carbonate (100\u2005mL), dried (MgSO4) and concentrated in\u2005vacuo to give 14 as an oil . LC\u2013MS: Rf 2.18 (+ESI) m/z 320.5 , Rf 2.20 (\u2212ESI) m/z 318.4 ([M\u2212H]\u2212); 1H\u2005NMR : \u03b4H=7.04 , 6.84 , 6.46 , 4.64 , 4.40\u20134.00 , 3.82 , 2.90\u20132.60 , 2.20\u20131.80 , and 1.30\u20131.20 . Characterization data for this compound are in complete agreement with an internal GE publication.boxylate . The chlH\u2010carbazole\u20104\u2010carboxylic acid (15)9\u2010(2\u2010Fluoroethyl)\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101. The ester 14 was dissolved in ethanol (10\u2005mL). A solution of sodium hydroxide dissolved in 10\u2005mL of water, was added. The reaction mixture was heated at reflux for 18\u2005h. The solvent was removed in\u2005vacuo and the crude mixture diluted with water (30\u2005mL), acidified with 2\u2009m HCl dropwise until acidic, and washed with CH2Cl2 (50\u2005mL). The organic layers were combined and dried (MgSO4) and concentrated in\u2005vacuo to give 15 , which was used crude into the next step. LC\u2013MS: Rf 1.81 (+ESI) m/z 292.4 ([M+H]+).H\u2010carbazole\u20104\u2010carbonyl chloride (16)9\u2010(2\u2010Fluoroethyl)\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101. A solution of 15 in anhydrous CH2Cl2 (5\u2005mL) was stirred under nitrogen. Oxalyl chloride was added followed by a drop of DMF. The reaction mixture was stirred at RT under nitrogen for 2\u2005h then evaporated in\u2005vacuo to give 16 as an oil, which was used into the next step without purification.NN\u2010ethyl\u20109\u2010(2\u2010fluoroethyl)\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101H\u2010carbazole\u20104\u2010carboxamide (GE387) (6)\u2010Benzyl\u2010. The acid chloride 16 was dissolved in anhydrous CH2Cl2 (8\u2005mL) and cooled to 0\u2009\u00b0C. N\u2010Ethylbenzylamine was then added and the reaction was stirred for 18\u2005h at RT. The reaction was quenched with 10\u2009% aqueous potassium carbonate (6\u2005mL). The CH2Cl2 layer was collected via a separatory funnel, dried (MgSO4), and then concentrated in\u2005vacuo. The crude material was purified by silica gel chromatography eluting with a gradient of 50\u2013100\u2009% EtOAc/petroleum ether (40\u201360\u2009\u00b0C) to afford the crude product as a yellow oil. The oil was then recrystallised from EtOAc to afford 6 as a pale\u2010yellow solid . LC\u2013MS: Rf 2.30 (+ESI) m/z 409.6 , Rf 2.35 (+ESI) m/z 409.6 ([M+H]+); 1H\u2005NMR : \u03b4H=7.42\u20137.21 , 7.02 , 6.84 , 6.43 , 4.80\u20134.40 , 4.31 , 3.62 , 3.83\u20133.41 2), 2.88\u20132.61 , 2.20\u20131.80 , and 1.32 2).N\u2010(2\u2010(Benzyloxy)ethyl)\u20102\u2010chloro\u20105\u2010methoxyaniline (18). Commercially available 9 was converted into the free base with 1\u2009m aqueous sodium carbonate (300\u2005mL). The mixture was extracted with CH2Cl2 (2\u00d7200\u2005mL), the organic layer dried (MgSO4) and evaporated to give an oil. Compound 9 was then dissolved in CH2Cl2 (50\u2005mL), in a dry flask under nitrogen. Benzyloxyacetaldehyde 17 and acetic acid were added. After 15\u2005min sodium triacetoxyborohydride was added. The mixture was stirred for 18\u2005h at RT and then poured into saturated aqueous ammonium chloride solution (100\u2005mL) and extracted with CH2Cl2 (2\u00d7100\u2005mL). The combined organic layers were dried (MgSO4) and evaporated to give an oil. The crude product was purified via silica flash chromatography eluting with CH2Cl2 to yield 18 as an oil . Material could not be obtained in a pure form, but regardless, the reaction was progressed to the next step. LC\u2013MS: Rf 2.42 (+ESI) m/z 292.3 ([M+H]+); 1H\u2005NMR : \u03b4H=7.37\u20137.30 , 7.14 CHCH), 6.23\u20136.18 CHCH), 4.71 , 3.74 , 3.73 , and 3.35 . Characterization data for this compound are in complete agreement with an internal GE publication.Ethyl 3\u2010((2\u2010(benzyloxy)ethyl)(2\u2010chloro\u20105\u2010methoxyphenyl)amino)\u20102\u2010hydroxycyclohex\u20101\u2010ene\u20101\u2010carboxylate (19). The aniline 18 was stirred in dry THF (150\u2005mL) at \u221240\u2009\u00b0C under nitrogen and potassium bis(trimethylsilyl) amide was added over 30\u2005min. The carboxylate 11 in dry THF (50\u2005mL) was then added and allowed to warm to RT over a period of 1.5\u2005h. Acetic acid was added (15\u2005mL) and concentrated in\u2005vacuo to remove the THF. EtOAc (200\u2005mL) and 10\u2009% aqueous potassium carbonate (200\u2005mL) was added and the mixture vigorously shaken. The EtOAc solution was separated, dried over (MgSO4) and concentrated in\u2005vacuo to afford 19 as an oil , which was used crude in the next step. LC\u2013MS: Rf 2.39 (\u2212ESI) m/z 458.5 ([M\u2212H]\u2212), Rf 2.53 (+ESI) m/z 460.5 ([M+H]+), Rf 2.39 (+ESI) m/z 460.5 ([M+H]+). Characterization data for this compound are in complete agreement with an internal GE publication.H\u2010carbazole\u20104\u2010carboxylate (20)Ethyl 9\u2010(2\u2010(benzyloxy)ethyl)\u20108\u2010chloro\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101. Zinc chloride was added to carboxylate 19 in dry Et2O (400\u2005mL) under nitrogen and heated at reflux for 5.5\u2005h. As the reaction was held at reflux, a thick brown dense oil formed in the reaction. The reaction was then cooled and the supernatant Et2O decanted off, EtOAc (300\u2005mL) was added, washed with 2\u2009m HCl (150\u2005mL) and with 10\u2009% aqueous potassium carbonate (150\u2005mL). The EtOAc layer was separated, dried (MgSO4) and concentrated in\u2005vacuo to afford an oil. The crude material was purified by silica gel chromatography eluting with a gradient of 10\u201340\u2009% EtOAc/petroleum ether (40\u201360\u2009\u00b0C) to afford 20 as an oil . The thick dense brown layer was treated with EtOAc (300\u2005mL) and 2\u2009m HCl (150\u2005mL). The EtOAc layer was separated, washed with 10\u2009% aqueous potassium carbonate (150\u2005mL), dried (MgSO4) and concentrated in\u2005vacuo to give an oil. Et2O (400\u2005mL) and anhydrous zinc chloride were added. The mixture was heated at reflux for a further 5\u2005days. The Et2O layer and the dark gum were both diluted with EtOAc (200\u2005mL) and then washed with 2\u2009m HCl (150\u2005mL), dried (MgSO4) and concentrated in\u2005vacuo to give a gum. This gum was purified by silica gel chromatography eluting with a gradient of 5\u201335\u2009% EtOAc/petroleum ether (40\u201360\u2009\u00b0C) to afford 20 as an oil . LC\u2013MS: Rf 2.66 (+ESI) m/z 442.5 ([M+H]+); 1H\u2005NMR : \u03b4H=7.31\u20137.16 , 6.93 , 6.34 , 4.58 , 4.40 , 4.20\u20134.09 , 3.86\u20133.80 , 3.79 , 2.82\u20132.63 , 2.10\u20131.80 , and 1.23 . Characterization data for this compound are in complete agreement with an internal GE publication.H\u2010carbazole\u20104\u2010carboxylic acid (21)9\u2010(2\u2010(Benzyloxy)ethyl)\u20108\u2010chloro\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101. To the ester 20 in ethanol (12\u2005mL) was added sodium hydroxide in water (1\u2005mL) and heated at 80\u2009\u00b0C for 18\u2005h. The ethanol was then removed by evaporation in\u2005vacuo and the residue partitioned between Et2O (30\u2005mL) and water (30\u2005mL). The Et2O layer was separated, dried (MgSO4) and concentrated in\u2005vacuo to give a gum. The aqueous layer was acidified to pH\u20051 with 2\u2009m HCl dropwise and extracted with CH2Cl2 (3\u00d720\u2005mL). The organic layers were dried (MgSO4) and concentrated in\u2005vacuo to afford 21 as a solid . LC\u2013MS: Rf 2.34 (+ESI) m/z 414.4 ([M+H]+); 1H\u2005NMR : \u03b4H=7.31\u20137.15 , 6.97 , 6.41 , 4.68\u20134.52 , 4.40 , 4.18 , 3.88 , 3.82 , 2.86\u20132.63 , and 2.27\u20131.86 . Characterization data for this compound are in complete agreement with an internal GE publication.NN\u2010ethyl\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101H\u2010carbazole\u20104\u2010carboxamide (22)\u2010Benzyl\u20109\u2010(2\u2010(benzyloxy)ethyl)\u20108\u2010chloro\u2010. The acid 21 was dissolved in THF (135\u2005mL) under N2 at RT. The solution was allowed to cool to 0\u2009\u00b0C and N\u2010ethylbenzylamine was added followed by 1\u2010hydroxybenzotriazole hydrate and N\u2010(3\u2010dimethylaminopropyl)\u2010N\u2032\u2010ethylcarbodiimide hydrochloride . Finally, triethylamine was added via syringe and the mixture stirred under N2 and warmed to RT over 48\u2005h. The reaction was then diluted with EtOAc (100\u2005mL) and filtered through Celite. The Celite was washed with more EtOAc (2\u00d7100\u2005mL). The combined filtrates were washed with 1\u2009m aqueous HCl (2\u00d7150\u2005mL), H2O (2\u00d7150\u2005mL), brine (150\u2005mL), dried (MgSO4) and concentrated in\u2005vacuo to give the product 22 as an oil , which was used crude in the next step. LC\u2013MS: Rf 2.74 (+ESI) m/z 531.6 ([M+H]+).NN\u2010ethyl\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101H\u2010carbazole\u20104\u2010carboxamide (23)\u2010Benzyl\u20109\u2010(2\u2010(benzyloxy)ethyl)\u2010. The carboxamide 22 in methanol (120\u2005mL) was shaken with 10\u2009% palladium on charcoal (1.5\u2005g), triethylamine and stirred for 18\u2005h at RT after purging twice (over 2\u2005h) with hydrogen gas under atmospheric pressure. The reaction was then filtered through a pad of Celite under nitrogen atmosphere and the filtrate concentrated in\u2005vacuo. The concentrate was then taken up in CH2Cl2 (200\u2005mL) and washed with 5\u2009% aqueous potassium carbonate solution (200\u2005mL). The CH2Cl2 solution was then separated, dried (MgSO4) and concentrated in\u2005vacuo to afford 23 as an oil , which was used crude in the next step. LC\u2013MS: Rf 2.62 (+ESI) m/z 497.5 ([M+H]+).NN\u2010ethyl\u20109\u2010(2\u2010hydroxyethyl)\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101H\u2010carbazole\u20104\u2010carboxamide (24)\u2010Benzyl\u2010. The benzyl\u2010protected intermediate 23 in methanol (50\u2005mL) was shaken with 10\u2009% palladium on charcoal (0.333\u2005g) and stirred for 18\u2005h at RT after purging twice (over 2\u2005h) with hydrogen gas under atmospheric pressure. The reaction was then filtered through a pad of Celite under nitrogen atmosphere and the filtrate concentrated in\u2005vacuo to give 24 as an oil , which was used crude in the next step. LC\u2013MS: Rf 2.08 (+ESI) m/z 407.5 ([M+H]+).H\u2010carbazol\u20109\u2010yl)ethyl4\u2010methylbenzenesulfonate (25)2\u2010(4\u2010(Benzyl(ethyl)carbamoyl)\u20105\u2010methoxy\u20101,2,3,4\u2010tetrahydro\u20109. The alcohol 24 was dissolved in anhydrous pyridine (5\u2005mL) under N2. The solution was cooled to 0\u2009\u00b0C and 4\u2010toluenesulfonyl chloride added potion\u2010wise to the solution over 30\u2005min. 4\u2010Dimethylaminopyridine was then added, and the solution was stirred and warmed to RT for 18\u2005h. The reaction was then quenched by careful addition of ice followed by water (5\u2005mL). The mixture was then extracted into EtOAc and washed with water (3\u00d710\u2005mL). Excess pyridine was removed by washing with 1\u2009m HCl (2\u00d710\u2005mL) and aqueous copper sulfate (2\u00d710\u2005mL). Excess 4\u2010toluenesulfonyl chloride was removed by washing with 1\u2009m aqueous sodium carbonate (2\u00d710\u2005mL). The organic layer was washed with brine (10\u2005mL), dried (MgSO4) and concentrated in\u2005vacuo. The crude product was recrystallised from EtOAc to afford 25 as an off\u2010white solid . LC\u2013MS: Rf 2.46 (+ESI) m/z 561.6 ([M+H]+); 1H\u2005NMR : \u03b4H=7.51 (CH2)CH3), 7.35\u20137.15 , 7.09 (CH2)CH3), 6.91 , 6.62 , 6.37 , 4.70\u20134.50 , 4.30\u20134.15 , 3.61 , 3.80\u20133.40 2), 2.80\u20132.50 , 2.33 , 2.1\u20131.60 , and 1.30 .H\u2010carbazol\u20109\u2010yl)ethyl methanesulfonate (26)2\u2010(4\u2010(Benzyl(ethyl)carbamoyl)\u20105\u2010methoxy\u20101,2,3,4\u2010tetrahydro\u20109. The phenol 24 in CH2Cl2 (20\u2005mL) was cooled to 0\u2009\u00b0C and methanesulfonyl chloride and triethylamine were added and allowed to warm to RT for 18\u2005h. The reaction was diluted with CH2Cl2 (20\u2005mL) washed with 5\u2009% aqueous potassium carbonate solution (40\u2005mL). The layers were separated. The combined organic layers were dried (MgSO4) and concentrated in\u2005vacuo to give a gum. The crude material was purified by silica gel flash chromatography eluting with a gradient of 50\u2013100\u2009% EtOAc/petroleum ether (40\u201360\u2009\u00b0C) to afford 26 as an oil . LC\u2013MS: Rf 2.12 (+ESI) m/z 485.5 ([M+H]+). 1H\u2005NMR : \u03b4H=7.40\u20137.10 , 7.03 , 6.86 , 6.44 , 4.65\u20134.50 , 4.50\u20134.45 , 3.64 , 3.90\u20133.20 2), 2.90\u20132.60 , 2.51 , 2.30\u20131.70 , and 1.31 .R,2S,5R)\u20102\u2010Isopropyl\u20105\u2010methylcyclohexyl 9\u2010(2\u2010(benzyloxy)ethyl)\u20108\u2010chloro\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101H\u2010carbazole\u20104\u2010carboxylate (29) and l\u2010menthol in CH2Cl2 (40\u2005mL) at 0\u2009\u00b0C was added N\u2010(3\u2010Dimethylaminopropyl)\u2010N\u2032\u2010ethylcarbodiimide hydrochloride and 4\u2010Dimethylaminopyridine . Stirring was continued at RT for 16\u2005h. After this period, H2O (100\u2005mL) and CH2Cl2 (100\u2005mL) were added. The organic layer was washed with 1\u2009m HCl, saturated NaHCO3 solution, and brine. The solution was dried (MgSO4), filtered, and concentrated. The crude racemic mixture 29 was separated into two diasteriomerically enriched samples via silica flash chromatography eluting with 30\u2009% Et2O/cyclohexane. LC\u2013MS: Rf 3.12 (+ESI) m/z 368.3 ; 1H\u2005NMR : \u03b4H=7.36\u20137.12 , 6.94 , 6.31 , 4.90 , 4.38 , 4.09 , 3.86\u20133.77 , 3.75 , 2.7\u20132.44 , 2.15\u20132.01 , 2.00\u20131.82 , 1.81\u20131.56 , 1.46\u20131.40 2, and 1.02\u20130.83 .R)\u20101\u2010Phenylprop\u20102\u2010yn\u20101\u2010yl 9\u2010(2\u2010(benzyloxy)ethyl)\u20108\u2010chloro\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101H\u2010carbazole\u20104\u2010carboxylate (30), N\u2010(3\u2010Dimethylaminopropyl)\u2010N\u2032\u2010ethylcarbodiimide hydrochloride and 4\u2010Dimethylaminopyridine in CH2Cl2 (5\u2005mL) was added (R)\u20101\u2010phenyl\u20102\u2010propyn\u20101\u2010ol at RT. The reaction was stirred for 4\u2005h at RT. The reaction mixture was diluted with CH2Cl2 (20\u2005mL) and washed with H2O (5\u2005mL). The organic layer was dried (MgSO4) and concentrated. The crude mixture was purified via silica flash column chromatography eluting with a gradient of 20\u201340\u2009% Et2O/petroleum ether (40\u201360\u2009\u00b0C) to yield 30 as a racemate . Separation of the two diastereomer was done via silica flash column chromatography eluting with a gradient of 10\u201320\u2009% Et2O/cyclohexane. LC\u2013MS: Rf 2.72 (+ESI) m/z 528.5 , 2.73 (+ESI) m/z 528.5 ; 1H\u2005NMR : \u03b4H=7.60\u20137.14 , 6.90 , 6.25 , 4.71\u20134.53 , 4.39 , 4.15 , 3.81 , 3.40 , 2.85\u20132.51 , 2.60 , and 2.20\u20131.79 .NN\u2010ethyl\u20109\u2010(2\u2010fluoroethyl)\u20105\u2010methoxy\u20102,3,4,9\u2010tetrahydro\u20101H\u2010carbazole\u20104\u2010carboxamide (6)\u2010Benzyl\u2010. 49.8\u2005mg of a racemic mixture of 6 was sent to Reach Separations to be separated into the two enantiomers. They provided the following methods for separation and purification. The racemate 6 was dissolved to 12\u2005mg\u2009mL\u22121 in MeOH/CH2Cl2 (1:1) and was then purified by SFC using a Lux C1 column at 40\u2009\u00b0C and eluting with isocratic 50:50 MeOH/CO2 with 0.2\u2009% v/v NH3 at a flow rate of 50\u2005mL\u2009min\u22121. The injection volume was 1500\u2005\u03bcL and peaks were detected at 223\u2005nm. Combined fractions of each of the two enantiomers were then evaporated to near dryness using a rotary evaporator, transferred into final vessels with CH2Cl2, which was removed under a stream of compressed air at 40\u2009\u00b0C before being stored in a vacuum oven at 40\u2009\u00b0C and 5\u2005mbar for 4\u2005h to afford the two enantiomers as yellow gums. Chiral purity of the enantiomers was analysed using a Lux C1 column at 40\u2009\u00b0C and eluting with isocratic 50:50 MeOH/CO2 with 0.2\u2009% v/v NH3 at a flow rate of 4\u2005mL\u2009min\u22121. The injection volume was 1.0\u2005\u03bcL and the peaks were detected at 210\u2013400\u2005nm. Chemical purity of the enantiomers was analysed using an Acquity BEH C18 column at 40\u2009\u00b0C and eluting with 0\u20134\u2005min gradient of 5\u201395\u2009% MeCN/H2O with 0.1\u2009% TFA, 4\u20104.02\u2005min gradient of 95\u2013100\u2009% MeCN/H2O with 0.1\u2009% TFA, 4.03\u2005min\u20134.5\u2005min isocratic 100\u2009% MeCN with 0.1\u2009% TFA, 4.5\u20134.52\u2005min gradient of 100\u20135\u2009% MeCN/H2O with 0.1\u2009% TFA, and 4.53\u20136\u2005min isocratic of 5\u2009% MeCN/H2O with 0.1\u2009% TFA at flow rate of 0.6\u2005mL\u2009min\u22121. The injection volume was 1.0\u2005\u03bcL and the peak was detected at 220\u2005nm. The final yield is 23.9\u2005mg of enantiomer (S)\u20106 with 98.02\u2009% chemical purity and in 100\u2009%\u2005ee, and 22.1\u2005mg of enantiomer (R)\u20106 with 100\u2009% chemical purity and in 98.8\u2009%\u2005ee.H\u2010carbazol\u20109\u2010yl)ethyl 4\u2010methylbenzenesulfonate (25)2\u2010(4\u2010(Benzyl(ethyl)carbamoyl)\u20105\u2010methoxy\u20101,2,3,4\u2010tetrahydro\u20109. 162.4\u2005mg of a racemic mixture of 25 was sent to Reach Separations to be separated into the two enantiomers. They provided the following methods for separation and purification. The racemate 25 was dissolved to 23\u2005mg\u2009mL\u22121 in MeOH/CH2Cl2 (1:1) and was then purified by SFC using a Lux C1 column at 40\u2009\u00b0C and eluting with isocratic 50:50 MeOH/CO2 with 0.2\u2009% v/v NH3 at a flow rate of 50\u2005mL\u2009min\u22121. The injection volume was 1000\u2005\u03bcL and peaks were detected at 223\u2005nm. Combined fractions of each of the two enantiomers were then evaporated to near dryness using a rotary evaporator, transferred into final vessels with CH2Cl2, which was removed under a stream of compressed air at 40\u2009\u00b0C before being stored in a vacuum oven at 40\u2009\u00b0C and 5\u2005mbar for 4\u2005h to afford the two enantiomers as white solids. Chiral purity of the enantiomers was analysed using a Lux C1 column at 40\u2009\u00b0C and eluting with isocratic 50:50 MeOH/CO2 with 0.2\u2009% v/v NH3 at a flow rate of 4\u2005mL\u2009min\u22121. The injection volume was 1.0\u2005\u03bcL and the peaks were detected at 210\u2013400\u2005nm. Chemical purity of the enantiomers was analysed using an Acquity BEH C18 column at 40\u2009\u00b0C and eluting with 0\u20134\u2005min gradient of 5\u201395\u2009% MeCN/H2O with 0.1\u2009% TFA, 4\u20104.02\u2005min gradient of 95\u2013100\u2009% MeCN/H2O with 0.1\u2009% TFA, 4.03\u2005min\u20134.5\u2005min isocratic 100\u2009% MeCN with 0.1\u2009% TFA, 4.5\u20134.52\u2005min gradient of 100\u20105\u2009% MeCN/H2O with 0.1\u2009% TFA, and 4.53\u20136\u2005min isocratic of 5\u2009% MeCN/H2O with 0.1\u2009% TFA at flow rate 0.6\u2005mL\u2009min\u22121. The injection volume was 1.0\u2005\u03bcL and the peak was detected at 220\u2005nm. The final yield is 82.7\u2005mg of enantiomer (S)\u201025 with 97.42\u2009% chemical purity and in 98.2\u2009%\u2005ee, and 77.1\u2005mg of enantiomer (R)\u201025 with 98.75\u2009% chemical purity and in 99.6\u2009%\u2005ee.m Tris\u22c5HCl, pH\u20057.4) were incubated with \u2248Kd concentration of [3H]1 and test compounds (0.3\u2005nm\u201310\u2005\u03bcm) at 4\u2009\u00b0C for 90\u2005min. Reactions were terminated by filtration through a 96\u2010well glass\u2010fibre filter plate (Millipore), and washed eight times with ice\u2010cold 50\u2005mm Tris\u22c5HCl. Microscint\u20050 was added to the dry filters, and radioactivity read in a Microbeta2 2450 Microplate Counter (PerkinElmer). Data were analysed using GraphPad Prism 6.0, and a four\u2010parameter nonlinear regression curve fit was used to calculate Ki values. Data are expressed as the mean\u00b1SEM from at least three independent experiments.Binding affinities were measured as per our published protocol.Circular dichroism spectroscopy. Solutions of (S)\u20106, (R)\u20106, and (S)\u20105 were prepared in concentrations of 0.1\u20130.2\u2005mg\u2009mL\u22121 in acetonitrile and CD spectra were collected on an Aviv 410 instrument.Radioisotope production. No\u2010carrier\u2010added aqueous [18F]fluoride ion was produced on a GE PETtrace cyclotron by irradiation of a 2.3\u2010mL silver\u2010bodied water target with a 25\u2010mA current and a 16.5\u2010MeV proton beam on 95\u2009% enriched 18O\u2010H2O via the nuclear 18O18F reaction.18F]KF\u2010Kryptofix\u2010222 complexPreparation of the [. [18F]Fluoride in [18O]H2O was transferred and immediately trapped on an anion\u2010exchange resin (Waters Sep\u2010Pak Accell Light QMA cartridge in the carbonate form) under vacuum. Trapped 18F\u2010fluoride was eluted from the Sep\u2010Pak cartridge and transferred to the reaction vessel with an eluent solution containing 0.25\u2009% wild\u2010type Kryptofix\u2010222\u00ae solution (1\u2005mL) in basic (0.05\u2009% wild\u2010type K2CO3) aq. MeCN (75\u2009% v/v). The solvents were evaporated in\u2005vacuo (130\u2005mbar) with a stream of N2 gas at 95\u2009\u00b0C over 5\u2005min. Anhydrous MeCN (3\u00d70.7\u2005mL) was then added and the mixture was azeotropically dried in\u2005vacuo (130\u2005mbar) with a stream of N2 at 95\u2009\u00b0C.18F]6Manual preparation of fluoride solution (125\u2005\u03bcL) of equivalent solvent. The vials were heated in a heating block to the desired temperature and a single vial was removed at each time point and quenched with 1:1 MeCN/H2O (300\u2005\u03bcL) before analyzing directly by HPLC using a gradient method: (pre\u2010injection) 1\u2005min equilibration at 10:90 MeCN/H2O. (post\u2010injection) 1\u2005min isocratic at 10:90 MeCN/H2O, 2\u2005min gradient to 70:30 MeCN/H2O, 3\u2005min isocratic at 70:30 MeCN/H2O. Flow rate: 1.5\u2005mL\u2009min\u22121. Run time: 6\u2005min (7\u2005min including equilibration time). Column: ACE\u00ae UltraCore, Super C18, 2.5\u2005\u03bcm, 50\u00d74.6\u2005mm; Serial No. A133239. A portion of the sample from 20\u2005min was taken and used for co\u2010injection with reference compound 6 to confirm synthesis of desired product.18F]6Automated preparation and formulation of KF\u2010Kryptofix\u2010222 complex and heated at 100\u2009\u00b0C for 20\u2005min. The reaction mixture was then cooled with compressed air and diluted with anhydrous MeCN (1\u2005mL). The crude mixture was injected into an ACE 160433 C18 5\u2005\u03bcm (100\u00d710\u2005mm) semipreparative reversed\u2010phase HPLC column. With a mobile phase of 48\u2009% aq. MeCN at a flow rate of 5\u2005mL\u2009min\u22121. [18F]6 was collected (tR: 19.6\u2005min) and immediately diluted with H2O (10\u2005mL). The aqueous solution was passed through a C18 cartridge . The cartridge was washed with H2O (15\u2005mL) and the radiolabelled product was eluted with EtOH (0.5\u2005mL). The radiolabelled product was then formulated with 0.9\u2009% w/v aq. NaCl in a vial to afford the radiolabelled title compound.18F]6Quality control for 6 at a tR of 9.8\u2005min. The area of the UV absorbance peak at 220\u2005nm corresponding to the carrier product was measured (integrated) on the HPLC chromatogram and compared with a standard curve relating mass to UV absorbance. The identity of the radiolabelled product was confirmed by HPLC co\u2010injection of the reference compound 6.n=2; weight: 230 and 302\u2005g) were obtained from Charles River Laboratories, UK. The rats were housed in Techniplast 2000P IVC cages on a layer of Aspen bedding in a room with constant temperature (21\u00b12\u2009\u00b0C) and fixed 12\u2005h light\u2013dark regime (lights on at 7:00\u2005am). Food and water were available ad libitum. After arrival, the rats were allowed to acclimatise for at least seven days. This research was regulated under the Animals (Scientific Procedures) Act 1986 Amendment Regulations 2012 following ethical review by the University of Cambridge Animal Welfare and Ethical Review Body (AWERB).Male Wistar rats .68Ge point source was made for attenuation and scatter correction of 511\u2005keV photons. The radiotracer, formulated in saline, was injected to the rats and the emission scan was started with tracer injection. A list\u2010mode protocol was used with an acquisition time of 60\u2005min.Prior to scanning, the animals were anaesthetised using isofluorane at a concentration of 5\u2009% in OImage reconstructions were performed using microPET Manager 2.4.1.1, ASIPro 6.7.1.2 (Siemens). Acquisition data were then Fourier re\u2010binned in 22 time frames and the data were reconstructed per timeframe employing an iterative reconstruction algorithm . The final datasets consisted of 95 slices with a slice thickness of 0.8\u2005mm, and an in\u2010plane image matrix of 128\u00d7128 pixels. Voxel size was 0.8\u00d70.8\u00d70.8\u2005mm. Data sets were corrected for decay, random coincidences, scatter and attenuation.\u22121 for brain tissue.Three\u2010dimensional regions of interest (ROIs) were drawn over the whole brain on an MRI template using PMOD software . PET images were then co\u2010registered with this MRI template and the regions of interest transferred from MRI to PET. Whole\u2010brain TACs were then obtained for each of the animals. The results were expressed as dimensionless standardised uptake values (SUV=[(tissue activity)\u00d7(body weight)]/injected dose). SUVs were calculated assuming a specific gravity of 1\u2005g\u2009mLThe authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re\u2010organized for online delivery, but are not copy\u2010edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file."} +{"text": "The propynyl substituent is nearly perpendicular to the plane formed by the rails of the boat. In the crystal, inversion dimers are formed by weak C\u2014H\u22efF hydrogen bonds with the dimers forming oblique stacks along the 18H12FNOS, is built up from a 4-fluoro\u00adbenzyl\u00adidene moiety and a di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit with a propynyl substituent, with the heterocyclic portion of the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit adopting a shallow boat conformation with the propynyl substituent nearly perpendicular to it. The two benzene rings are oriented at a dihedral angle of 43.02\u2005(6)\u00b0. In the crystal, C\u2014HFlurphen\u22efFFlurphen (Flurphen = fluoro\u00adphen\u00adyl) hydrogen bonds link the mol\u00adecules into inversion dimers, enclosing R22(8) ring motifs, with the dimers forming oblique stacks along the a-axis direction. Hirshfeld surface analysis of the crystal structure indicates that the most important contributions to the crystal packing are from H\u22efH (33.9%), H\u22efC/C\u22efH (26.7%), H\u22efF/F\u22efH (10.9%) and C\u22efC (10.6%) inter\u00adactions. Hydrogen bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Density functional theory (DFT) optimized structures at the B3LYP/6\u2013311\u2005G level are compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.The title compound, C The propynyl substituent is nearly perpendicular to the plane defined by C1, C6, C7 and C8, as shown by the C6\u2014N1\u2014C9\u2014C10 torsion angle of 81.3\u2005(2)\u00b0. A puckering analysis of the heterocyclic ring of the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit shows that it adopts a shallow boat conformation with puckering parameters QT = 0.3759\u2005(14)\u2005\u00c5, q2 = 0.3639\u2005(15)\u2005\u00c5, q3 = \u22120.0938\u2005(17)\u2005\u00c5, \u03c6 = 173.6\u2005(3)\u00b0 and \u03b8 = 104.5\u2005(3)\u00b0. In the heterocyclic ring B, the C1\u2014S1\u2014C8 [101.73\u2005(8)\u00b0], S1\u2014C8\u2014C7 [119.93\u2005(12)\u00b0], C8\u2014C7\u2014N1 [119.23\u2005(14)\u00b0], C7\u2014N1\u2014C6 [125.59\u2005(14)\u00b0] and C6\u2014C1\u2014S1 [122.07\u2005(13)\u00b0] bond angles are enlarged, while the N1\u2014C6\u2014C1 [120.91\u2005(15)\u00b0] bond angle is narrowed when compared with the corresponding values in the closely related compounds 4-methyl-3,4-di\u00adhydro-2H-1,4-benzo\u00adthia\u00adzin-3-one, (II) -one, (III) -2-(2-chloro\u00adbenzyl\u00adidene)-4-(prop-2-yn\u00adyl)-2H-1,4-benzo\u00adthia\u00adzin-3(4H)-one, (IV), Flurphen\u22efFFlurphen (Flurphen = fluoro\u00adphen\u00adyl) hydrogen bonds analysis have a nearly symmetrical distribution of points, Fig.\u00a07c, with the thick edges at de + di \u223c2.70\u2005\u00c5. The pair of characteristic wings in the fingerprint plot delineated into H\u22efF/F\u22efH contacts arises from the C\u2014H\u22efF hydrogen bonds have an arrow-shaped distribution of points with the tip at de = di \u223c1.68\u2005\u00c5. The pair of characteristic wings in the fingerprint plot delineated into H\u22efO/O\u22efH contacts have a pair of spikes with the tips at de + di = 2.54\u2005\u00c5. Finally, the H\u22efS/S\u22efH contacts are viewed as A pair of wide spikes with the tips at de + di = 3.02\u2005\u00c5. The Hirshfeld surface representations with the function dnorm plotted onto the surface are shown for the H\u22efH, H\u22efC/C\u22efH, H\u22efF/F\u22efH, C\u22efC, H\u22efO/O\u22efH and H\u22efS/S\u22efH inter\u00adactions in Fig.\u00a08a\u2013f, respectively.The overall two-dimensional fingerprint plot, Fig.\u00a07s Table\u00a01 as well s Table\u00a01 and is set al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing using standard B3LYP functional and 6\u2013311G basis-set calculations -2-(4-fluoro\u00adbenzyl\u00adidene)-4-(prop-2-yn\u00adyl)-2H-1,4-benzo\u00adthia\u00adzin-3(4H)-one ring. The energy band gap [\u0394E = ELUMO - EHOMO] of the mol\u00adecule was about 3.92\u2005eV, and the frontier mol\u00adecular orbital energies, EHOMO and ELUMO were \u22125.85 and \u22121.93\u2005eV, respectively.The optimized structure of the title compound, (I)II in the Cambridge Crystallographic Database to 36\u00b0 (IId). The other three have the benzo\u00adthia\u00adzine unit nearly planar with the corresponding dihedral angle of ca 3\u20134\u00b0. In the case of IIa, the displacement ellipsoid for the sulfur atom shows a considerable elongation perpendicular to the mean plane of the heterocyclic ring, suggesting disorder, and a greater degree of non-planarity but for the other two, there is no obvious source for the near planarity.Using the search fragment Z)-2-(4-fluoro\u00adbenzyl\u00adidene)-2H-1,4-benzo\u00adthia\u00adzin-3(4H)-one (1.6\u2005mmol), potassium carbonate (4\u2005mmol) and tetra-n-butyl ammonium bromide (0.15\u2005mmol) in DMF (20\u2005ml). Stirring was continued at room temperature for 24\u2005h. The salts were removed by filtration and the filtrate was concentrated under reduced pressure. The residue was separated by chromatography on a column of silica gel with ethyl acetate\u2013hexane (2/8) as eluent. The solid product obtained was recrystallized from ethanol to afford colourless crystals (yield: 89%).Propargyl bromide (4\u2005mmol) was added to a mixture of I, global. DOI: 10.1107/S2056989019002354/lh5893Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019002354/lh5893Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019002354/lh5893Isup6.cmlSupporting information file. DOI: 1897371CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title hydrated azo dye, the benzene and thia\u00adzole make a dihedral angle of 4.69\u2005(17)\u00b0. In the crystal, hydrogen bonds, C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions resulting in the formation of a three-dimensional framework. 10H10N4OS\u00b7H2O, the benzene and thia\u00adzole, are nearly coplanar, with a dihedral angle between their mean planes of 4.69\u2005(17)\u00b0. The aromatic rings on the \u2013N=N\u2013 moiety exhibit a trans configuration. The crystal structure features many types of inter\u00admolecular inter\u00adactions involving all the functional groups \u2013 strong hydrogen bonds (N\u22efH and O\u22efH), weak hydrogen bonds (C\u2014H\u22efO and C\u2014H\u22efN), C\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions \u2013 resulting in the formation of a three-dimensional framework.In the title hydrated azo dye, C Azo dyes have wide range applications in the cosmetic, food, textile industry, chemical sensing, and pharmaceutical . The meth\u00adoxy and amino groups on the benzene ring are co-planar with the ring with atoms O1 and N4 deviating by \u22120.010\u2005(2) and \u22120.019\u2005(4)\u2005\u00c5, respectively. The dihedral angle between the thia\u00adzole and benzene rings is 4.69\u2005(17)\u00b0, nearly coplanar.The mol\u00adecular structure of (I)a, Table\u00a012), azo (\u2013N=N\u2013) and thia\u00adzole groups and the water mol\u00adecule of crystallization are the primary inter\u00adactions responsible for the formation of the three dimensional structure. In addition, the crystal structure is supported by other inter\u00admolecular inter\u00adactions as a secondary weak inter\u00adactions, C\u2014H\u22efX (X = O and N), C\u2014H\u22ef\u03c0 and offset \u03c0\u2014\u03c0 inter\u00adactions. The weak hydrogen bonds are formed between the C\u2014H moieties in the benzene and thia\u00adzole rings with amine, azo, meth\u00adoxy groups of adjacent mol\u00adecules and water mol\u00adecules . On the shape index surface , convex blue regions represent hydrogen-donor groups and concave red regions represent hydrogen-acceptor groups. In addition, concave red regions represent C\u2014H\u22ef\u03c0 and offset \u03c0\u2013\u03c0 inter\u00adactions. The amino group behaves as both a donor and an acceptor. The methyl part of the meth\u00adoxy group acts as a donor while the oxygen atom is an acceptor.Hirshfeld surfaces and fingerprint plots were generated using ce Fig.\u00a03b, conveet al., 2007The two-dimensional fingerprint plots Fig.\u00a04 qu\u00adantifet al., 1992trans configuration and the phenolic oxygen atom is linked to an azo nitro\u00adgen atom by intra\u00admolecular hydrogen bonding. The crystal structure features only van der Waals inter\u00adactions. To form complexes with metal ions, both thia\u00adzole and naphthol rings are rotated by 180\u00b0 to coordinate to the metal through the phenolic oxygen atom, the azo nitro\u00adgen atom adjacent to the naphthol ring and the thia\u00adzole nitro\u00adgen atom, resulting the formation of five-membered chelate rings. Complexes of TAR and TAC are formed in a similar way due to the presence of a hydroxyl group in the structure M HCl (16\u2005ml), and 8.236\u2005mmol of sodium nitrate solution was added slowly under stirring at low temperature 268\u2013273\u2005K until the diazo\u00adnium salt was obtained. m-Anisidine (1.12\u2005ml in 40\u2005ml of 4\u2005M HCl) was slowly dropped into the mixture and stirred at a temperature between 268 and 273\u2005K for 1\u2005h. After the reaction was complete, conc. NH3 was dropped into the mixture (pH 6) until the red\u2013orange crude produce appeared. The products were filtered, washed with cold water, purified by column chromatography and recrystallized from an aceto\u00adnitrile\u2013water (1:1) mixture by vapour diffusion.2-Amino\u00adthia\u00adzole (9.986\u2005mmol) was dissolved in 6\u20051H NMR : \u03b4 3.806 , 6.364 , 6.374 , 7.546 ,7.629 , 7.697 , 7.883 . Mass spectroscopy: m/z 235.0654 [C10H11N4OS+], 205.0548 [C9H9N4S.+], 150.0662 [C7H8N3O.+], 122.0601 [C7H8NO.+]. IR (KBr cm\u22121): 3,413\u2005cm\u22121 ; 821\u2005cm\u22121 ; 1,617 ; 1,222\u2005cm\u22121 ; 1,103\u2005cm\u22121 ; 1,152\u2005cm\u22121 ; 1,541cm\u22121 ; 1,021\u2005cm\u22121 . Elemental analysis calculated for C10H10N4OS\u00b7H2O: C, 51.27; H, 4.30; N, 23.92. Found: C, 51.34; H, 4.20; N, 23.98.Uiso = 1.2 Ueq(C-aromatic) and 1.5Ueq (C-meth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901900207X/dx2014sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S205698901900207X/dx2014Isup2.hklStructure factors: contains datablock(s) I. DOI: 1895710CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The structure has triclinic (P2(O2CCPh3-\u03bc-\u03baO:\u03baO\u2032)]2 is located about an inversion centre. The tri\u00adphenyl\u00adacetate ligand displays a \u03bc-\u03baO:\u03baO\u2032-bridging coordination mode, leading to the formation of an octa\u00adgonal Al2O4C2 core. The complex displays HPh\u22efCPh inter\u00admolecular inter\u00adactions.Single crystals of the title compound, [Al(iBu) The tri\u00adphenyl\u00adacetate ligand exhibits a \u03bc2-\u03baO:\u03baO\u2032-bridging coordination mode. The CPh\u2014CPh [1.3788\u2005(17)\u2005\u00c5 for C5\u2014C6 to 1.4016\u2005(14)\u2005\u00c5 for C15\u2014C16], CiBu\u2014CiBu [1.523\u2005(2)\u2005\u00c5 for C26\u2014C28 to 1.5381\u2005(16)\u2005\u00c5 for C21\u2014C22], C1\u2014C2 [1.5473\u2005(13)\u2005\u00c5] and C1\u2014Cipso [1.5425\u2005(14)\u2005\u00c5 for C2\u2014C9 to 1.5455\u2005(14)\u2005\u00c5 for C2\u2014C3] bond lengths inside the ligands are within the expected ranges. The complex has a nearly flat eight-membered Al2O4C2 core, with the greatest deviations from the plane being 0.0548\u2005(6)\u2005\u00c5 for the O2 and O2i atoms. The bond angles inside the core are 106.84\u2005(4) (O1\u2014Al\u2014O2i), 151.00\u2005(7) (Al\u2014O1\u2014C1), 123.64\u2005(9) (O1\u2014C1\u2014O2) and 156.79\u2005(8)\u00b0 (C1\u2014O2\u2014Ali), summing to a value of 1076.54\u00b0 for the entire core, which deviates from a flat octa\u00adgon by 3.46\u00b0. The Al\u2014X bond lengths are given in Table\u00a01R2(\u03bc-O2CR\u2032)]2 compounds (see \u00a74 below) having the Al2O4C2 core (23 independent core fragments) have Al\u2014X bond lengths varying from ca 1.77 to 1.86\u2005\u00c5 (average 1.82\u2005\u00c5) for Al\u2014O, 1.92\u20132.00\u2005\u00c5 (average 1.96\u2005\u00c5) for Al\u2014C and 1.23\u20131.29\u2005\u00c5 (average 1.26\u2005\u00c5) for C\u2014O bonds. The bond lengths in the title complex (Table\u00a012-bridging RO\u2212 ligands), but shorter than the Al\u2014O distances in complexes with either Al\u2013O=CR2 or Al\u2014Oether fragments due to different types of Al\u2014O inter\u00adactions, changing from the ion\u2013ion type in the case of Al\u2014Oalk\u00adyl/ar\u00adyl bonds to the ion\u2013dipole one in the case of Al\u2014O=CR2 or Al\u2014Oether fragments.The title compound crystallizes in the triclinic space group ]2 Fig.\u00a02 located x Table\u00a01 are closA\u22efH20 and 2.30\u2005\u00c5 for H25A\u22efH12. Two inter\u00admolecular inter\u00adactions involving aromatic H atoms with the \u03c0-system of a arene group have been found, i.e. 2.89\u2005\u00c5 for H6\u22efC12 and 2.98\u2005\u00c5 for H7\u22efC11 (see Table S1 for details). Inter\u00adacting arene rings are located nearly perpendicular to one another, with the corresponding angle between the C3\u2013C8 and C9\u2013C14 planes being 82.83\u2005(3)\u00b0 .The crystal lattice exhibits weak inter\u00admolecular van der Waals contacts between methyl or methyl\u00adene and aromatic H atoms, with the distances being 2.49\u2005\u00c5 for H23)\u00b0 Fig.\u00a03. The laset al., 20162O4C2 core and having the [AlR2(\u03bc-O2CR\u2032)]2 motif, where R is alkyl or C6F5. Records for crystal structures with other R groups connected to Al via the C atom have not been found in the CSD. 14 complexes have bridging carboxyl\u00adate ligands, the others have a heteroatom in the \u03b1-position .According to the Cambridge Structural Database ]2 type are represented by structures with R = R\u2032 = Me and aryl = Ph 3 and aryl = 4-MeC6H4 and R = isobutyl (iBu) ]2 complexes with a substituted acetate anion possess R = Et and CX3 = CPh3 ]2, with X = \u2013C4H(CH3)2Zr(\u03b75-C5Me5)2 2(\u03bc-O2CC6F5)]2 ]2 ]2, with R = iBu depends greatly on various inter\u00adactions within the complex, including nonvalence ones. See also related ab initio calculations in the literature 3(THF)3] was prepared according to a previously published method was added dropwise to a suspension of [Nd(Ph3CCOO)3(THF)3] in 15\u2005ml of hexane at room temperature. The suspension dissolved within a few minutes upon addition. The resulting solution was stirred overnight at room temperature. Crystals of [Al(iBu)2(Ph3CCOO)]2 were isolated from the reaction mixture by crystallization at 243\u2005K for 2\u2005d. The mother liquor was deca\u00adnted and crystals were dried under dynamic vacuum. The yield was 56\u2005mg . Calculated for C56H66Al2O4 (%): C 78.48, H 7.76; found: C 78.17, H 8.01.A solution of Al(iBu)Uiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) otherwise. A rotating group model was applied for methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019003396/tx2010sup1.cifCrystal structure: contains datablock(s) global. DOI: 10.1107/S2056989019003396/tx2010sup4.pdfC...H interactions and packing plots. DOI: 1902149CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal. hydroxyl-O\u2014H\u22efO(hydrox\u00adyl) and hydroxyl-O\u2014H\u22efS(thione) hydrogen bonds give rise to a supra\u00admolecular layer in the ab plane.The title tri-substituted thio\u00adurea derivative is twisted with a dihedral angle of 72.12\u2005(9)\u00b0 between the planes through the CN 13H18N2O3S, the thione-S and carbonyl-O atoms lie, to a first approximation, to the same side of the mol\u00adecule [the S\u2014C\u2014N\u2014C torsion angle is \u221249.3\u2005(2)\u00b0]. The CN2S plane is almost planar (r.m.s. deviation = 0.018\u2005\u00c5) with the hy\u00addroxy\u00adethyl groups lying to either side of this plane. One hy\u00addroxy\u00adethyl group is orientated towards the thio\u00adamide functionality enabling the formation of an intra\u00admolecular N\u2014H\u22efO hydrogen bond leading to an S(7) loop. The dihedral angle [72.12\u2005(9)\u00b0] between the planes through the CN2S atoms and the 4-tolyl ring indicates the mol\u00adecule is twisted. The experimental mol\u00adecular structure is close to the gas-phase, geometry-optimized structure calculated by DFT methods. In the mol\u00adecular packing, hydroxyl-O\u2014H\u22efO(hydrox\u00adyl) and hydroxyl-O\u2014H\u22efS(thione) hydrogen bonds lead to the formation of a supra\u00admolecular layer in the ab plane; no directional inter\u00adactions are found between layers. The influence of the specified supra\u00admolecular inter\u00adactions is apparent in the calculated Hirshfeld surfaces and these are shown to be attractive in non-covalent inter\u00adaction plots; the inter\u00adaction energies point to the important stabilization provided by directional O\u2014H\u22efO hydrogen bonds.In the title tri-substituted thio\u00adurea derivative, C R1(R2)NC(=S)N(R3)R4 for R1\u20134 = alk\u00adyl/aryl. The present study concerns a tri-substituted example, i.e. an N,N\u2032-di-N\u2032-benzoyl\u00adthio\u00adurea derivative, notable for having a carbonyl group connected to the thio\u00adurea framework. Thio\u00adurea mol\u00adecules are of inter\u00adest in themselves and as ligands for metal ions 2, (I)The amine-H atoms in thio\u00adurea, Hsyn as the S1\u2014C1\u2014N2\u2014C6 torsion angle is \u221249.3\u2005(2)\u00b0; the O3\u2014C6\u2014N2\u2014C1 torsion angle is \u22126.8\u2005(3)\u00b0. The hy\u00addroxy\u00adethyl groups lie to either side of the CN2S plane (r.m.s. deviation = 0.018\u2005\u00c5). The O1-hy\u00addroxy\u00adethyl group is folded toward the thio\u00adamide part of the mol\u00adecule, an orientation that allows for the formation of an intra\u00admolecular N2\u2014H\u22efO1 hydrogen bond that closes an S(7) loop, Table\u00a022S atoms and the terminal aryl ring. The C1\u2014N1 bond length is considerably shorter than the C1\u2014N2 bond, which suggests some delocalization of \u03c0-electron density over the S1\u2014C1\u2014N1 atoms that does not extend over the C1\u2014N1\u2014C6 atoms, consistent with the large twist about the C1\u2014N2 bond (see above). The bond angles subtended at the C1 and C6 atoms follow the expected trends in that those involving the formally doubly bonded atoms are wider, by approximately 10\u00b0, compared with the other angles, Table\u00a01The title compound, (I)d,p) basis set N1\u2014C2\u2014C3\u2014O1 and N1\u2014C4\u2014C5\u2014O2 torsion angles, which are disparate, by about 12\u00b0, in the experimental structure but are symmetric, i.e. \u00b169\u00b0, in the optimized structure.Compound (I)1 symmetry to generate helical chains along the b-axis direction. The O\u2014H\u22efS hydrogen bonding serves to connect translationally related chains along the a-axis direction and these contacts are reinforced by phenyl-C\u2014H\u22efO(carbon\u00adyl) inter\u00adactions. In this way, a supra\u00admolecular layer in the ab plane is formed, Fig.\u00a03a). Layers stack along the c-axis direction without directional inter\u00adactions between them, Fig.\u00a03b).In the crystal of (I)Crystal Explorer 17 for (I)dnorm in Fig.\u00a05O atoms, Table\u00a03The Hirshfeld surface mapped over electrostatic potential in Fig.\u00a04ER) descriptor, which is derived from the analysis of the Hirshfeld surface , for a pair of elements is defined as the ratio between proportion of actual contacts in the crystal to the theoretical proportion of random contacts. This ratio is greater than unity for a pair of elements having a high likelihood to form contacts in a crystal, while it is less than one for a pair which tends to avoid contacts with each other. A listing of ER values for (I)The inter\u00admolecular contacts in the crystal of (I)a)\u2013(e), respectively. A summary of the percentage contributions from the various contacts in the crystal are given in Table\u00a05b) and has an ER value of 0.92, i.e. close to unity. The contribution from O\u22efH/H\u22efO contacts is viewed as long spikes at de + di \u223c1.8\u2005\u00c5, with points scattered around different regions in the delineated fingerprint plot, Fig.\u00a06c). In the fingerprint delineated into C\u22efH/H\u22efC contacts in Fig.\u00a06d), a pair of small tips at de + di < 2.8\u2005\u00c5 is the result of short inter\u00adatomic contacts, Table\u00a03A\u22efO1). The percentage contribution from S\u22efH/H\u22efS contacts (13.1%) reflect the presence of O\u2014H\u22efS hydrogen bonds and are apparent through the appearance of asymmetric spikes at de + di \u223c2.1\u2005\u00c5 in Fig.\u00a06e).The overall fingerprint plots for (I)EBSSEint) listed in Table\u00a06\u22121 more stable than the O\u2014H\u22efS inter\u00adaction despite phenyl-C\u2014H being a weak hydrogen-bond donor and thione-S a weak acceptor, and that such inter\u00adactions are known to be dispersive in nature of all pairwise inter\u00adactions sums to \u221236.11 kcal mol\u22121, while the total dispersion energy term (Edispersion) computes to \u221243.83 kcal mol\u22121.As the mol\u00adecular packing is governed directionally by hydrogen bonding between mol\u00adecules, the energy frameworks were simulated (Turner 2CH2OH)2, (II), has been reported twice i.e. of the type phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl), to sustain a three-dimensional architecture. The other closely related structure is that of 4-MePhC(=O)N(H)C(=S)N(Me)CH2CH2OH) N(H)C(=S)N. The optical absorption spectra were measured on 10 and 100\u2005\u00b5M ethanol:aceto\u00adnitrile (1:1) solutions in the range 190\u20131100\u2005nm on a double-beam Shimadzu UV 3600 Plus UV\u2013vis spectrophotometer. The thermogravimetric analysis (TGA) was performed on a Perkin Elmer STA 6000 Simultaneous Thermogravimetric Analyzer in the range of 35\u2013900\u00b0C under a nitro\u00adgen atmosphere at a flow rate of 10\u00b0C min\u22121. The experimental powder X-ray diffraction pattern was measured on a Rigaku MiniFlex diffractometer with Cu K\u03b11 radiation (\u03bb = 1.54056\u2005\u00c5) in the 2\u03b8 range of 5\u201370\u00b0 and a step size of 0.02\u00b0. The experimental PXRD patterns were compared to the simulated PXRD patterns calculated from the CIF using the Rigaku PDXL structure analysis software package. The patterns matched indicating that the reported crystal structure is representative of the bulk material.All chemicals and solvents were used as purchased without purification. The reactions were carried out under ambient conditions. The melting point was measured using a Hanon MP-450 melting point apparatus. The CHN elemental analysis was performed on a LECO TruSpec Micro analyser under helium atmosphere with glycine being used as the standard. The IR spectrum was measured on a Bruker Vertex 70v FT\u2013IR spectrophotometer from 4000 to 400\u2005cm13H18N2O3S, found : C 55.59 (55.30), H 6.57 (6.43), N 9.79 (9.92). IR : 3312 , 3158 , 3061 , 2955\u20132881 , 1686 , 1539 , 1250 , 1054 , 747 . UV : \u03bbmax nm 354.4 , 294.0 , 246.4 , 202.6 . 1H NMR , 7.76 , 7.31 , 5.66 , 4.87 , 3.98 , 3.76 , 3.70 , 2.37 . 13C{1H} NMR : \u03b4 180.63 (C1), 163.78 (C6), 141.88 (C7), 130.13 (C10), 128.47 , 127.28 , 58.58 (C5), 56.95 (C3), 54.42 (C4), 54.29 (C2), 20.42 (C13).Synthesis of (I)2O between 135 and 165\u00b0C, which corresponds to approximate 6% of the weight for (I)The pyrolytic processes for (I)Uiso(H) set to 1.2\u20131.5Ueq(C). The O- and N-bound H atoms were located from a difference map and refined with O\u2014H and N\u2014H = 0.84\u00b10.01 and 0.88\u00b10.01\u2005\u00c5, respectively, and with Uiso(H) = 1.5Ueq(O) and 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S2056989019012581/hb7854sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019012581/hb7854Isup2.hklStructure factors: contains datablock(s) I. DOI: 1919878CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The second, C8H11N9O2S, contains two azide groups with an average N\u03b1\u2014N\u03b2 distance of 1.249\u2005(2)\u2005\u00c5 and an average N\u03b2\u2014N\u03b3 distance of 1.132\u2005(2)\u2005\u00c5. Each compound contains a bulky protecting group.The structures of two azide containing imidazole derivatives are reported. The first, C 8H12N6O2S, contains one azide group with an N\u03b1\u2014N\u03b2 distance of 1.229\u2005(2)\u2005\u00c5 and an N\u03b2\u2014N\u03b3 distance of 1.128\u2005(2)\u2005\u00c5. The second, C8H11N9O2S, contains two azide groups with an average N\u03b1\u2014N\u03b2 distance of 1.249\u2005(2)\u2005\u00c5 and an average N\u03b2\u2014N\u03b3 distance of 1.132\u2005(2)\u2005\u00c5. Each compound contains a bulky protecting group (di\u00admethyl\u00adamino\u00adsulfon\u00adyl) which can be easily removed under mildly acidic conditions.The structures of two azide containing imidazole derivatives are reported. Allylic azides are fairly reactive making them attractive starting compounds to convert into amides. The first, C These compounds were synthesized in the previous study but the structures were not reported. Figs. 11 and 2, respectively.The efficient synthesis of nagelamide alkaloids has garnered inter\u00adest \u2005\u00c5 and an N4\u2014N5 distance of 1.128\u2005(2)\u2005\u00c5. The N3\u2014N4\u2014N5 angle is 172.32\u2005(13)\u00b0. The azide groups in 2 show an N3\u2014N4 distance of 1.253\u2005(2)\u2005\u00c5, N4\u2014N5 distance of 1.129\u2005(2)\u2005\u00c5, N6\u2014N7 distance of 1.239\u2005(2)\u2005\u00c5, and N7\u2014N8 distance of 1.134\u2005(2)\u2005\u00c5. The N3\u2014N4\u2014N5 angle is 171.58\u2005(15)\u00b0 and the N6\u2014N7\u2014N8 angle is 173.95\u2005(15)\u00b0. All three azide moieties in both compounds show the same general trend of a longer N\u03b1\u2014N\u03b2 distance and shorter N\u03b2\u2014N\u03b3 distance with a quasilinear geometry. This is typical for covalent azides with terminal N\u03b2\u2014N\u03b3 demonstrating more triple-bond character. A previously reported covalent azide occurring in the compound ethyl-2-[(azido\u00adcarbon\u00adyl)amino]\u00adbenzoate demonstrated bond lengths N\u03b1\u2014N\u03b2 of 1.264\u2005(2)\u2005\u00c5 and N\u03b2\u2014N\u03b3 of 1.131\u2005(2)\u2005\u00c5 and an N\u03b1\u2014N\u03b2\u2014N\u03b1 angle of 174.7\u2005(2)\u00b0 in 1 was \u2212115.21\u2005(13)\u00b0 while the related torsion angle (C5\u2014C6\u2014N6\u2014N7) in 2 was 50.25\u2005(18)\u00b0. 2 contains one azide group bound to the imidazole at C2 and shows a torsion angle N1\u2014C2\u2014N3\u2014N4 of \u2212174.82\u2005(11)\u00b0. The allylic azides in both compounds exhibit a similar dihedral angle between the azide and the imidazole ring, 70.3\u2005(11)\u00b0 for 1 and 77.3\u2005(17)\u00b0 for 2. While the imidazole-bound azide in 2 shows a dihedral angle of 5.0\u2005(10)\u00b0. Indeed, the torsion angle and dihedral angle for this particular azide demonstrate the near planarity between the imidazole and its covalently bound azide. Figs. 31 and 2, respectively.The torsion angles for the azides and dihedral angles between the azides and imidazole rings for both compounds have been measured. The allylic azide torsion angles between sp3-hybrid\u00adized, as validated by the C\u2014N\u2014C bond angles C6\u2014N6\u2014C8 = 113.86\u2005(10)\u00b0 for 1 and C7\u2014N9\u2014C8 = 113.93\u2005(12)\u00b0 for 2. Both compounds also contain a double bond between C4 and C5. The measured bond distance is 1.333\u2005(2)\u2005\u00c5 for 1 and 1.340\u2005(2)\u2005\u00c5 for 2.Both title compounds contain a DMAS protecting group. The amine component of this protecting group is 1 is substituted at the N1 and C3 position with no substitution at C2. The N1\u2014C2 distance is 1.378\u2005(2)\u2005\u00c5 while the N2\u2014C2 distance is 1.301\u2005(2)\u2005\u00c5. However, in 2, the imidazole ring is substituted with an azide group at C2 but this seemingly has no effect on the ring bond distances. The measured bond distances for N1\u2014C2 and N2\u2014C2 in 2 are 1.385\u2005(2) and 1.310\u2005(2)\u2005\u00c5, respectively.The imidazole ring in 1 is 1.686\u2005(1)\u2005\u00c5 and 1.718\u2005(1)\u2005\u00c5 for 2. The disparity may be attributed to the presence of azide, which is substituted at the C2 position for 2.There is, however, a significant difference in the measured N1\u2014S1 distance for the two compounds. The imidazole ring is substituted at the N1 position for both compounds with DMAS. The N1\u2014S1 distance for 1 shows C1\u2014H1\u22efO1i and C2\u2014H2\u22efO2ii inter\u00adactions of 2.53 and 2.39\u2005\u00c5, respectively. There is also a C4\u2014H4\u22efN5iii inter\u00adaction of 2.70\u2005\u00c5 acrylate, was reported \u2005\u00c5 which is similar to the bond distances in 1 and 2 [1.333\u2005(2) and 1.340\u2005(2)\u2005\u00c5 respectively].A search of related compounds was conducted in the Cambridge Structural Database = 1.5Ueq(C) for methyl H and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details for 10.1107/S205698901900519X/zl2753sup1.cifCrystal structure: contains datablock(s) compound_1, compound_2. DOI: 10.1107/S205698901900519X/zl2753compound_1sup2.hklStructure factors: contains datablock(s) compound_1. DOI: 10.1107/S205698901900519X/zl2753compound_2sup3.hklStructure factors: contains datablock(s) compound_2. DOI: Click here for additional data file.10.1107/S205698901900519X/zl2753compound_1sup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901900519X/zl2753compound_2sup5.cmlSupporting information file. DOI: 1910313, 1910312CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "CavCOOH-in, was synthesized and crystallized with acetic acid to evaluate its mol\u00adecular recognition properties towards this analyte in the solid state.Acetic acid is a by-product of peracetic acid with acute irritant properties when present in air. The cavitand 5,11,17,23-tetra\u00admethyl-4,24:6,10:12,16:18,22-tetra\u00adkis\u00ad(methyl\u00adenedi\u00adoxy)resorcin[4]arene functionalized at the upper rim with a carb\u00adoxy\u00adlic acid group, CavCOOH-in, of chemical formula C37H32O10, was synthesized in order to study its supra\u00admolecular inter\u00adactions with acetic acid in the solid state. Crystals suitable for X-ray diffraction analysis were obtained by slow evaporation of a di\u00adchloro\u00admethane\u2013acetone solution of CavCOOH-in, to which glacial acetic acid had been added. The resulting compound, C37H32O10\u00b72C2H4O2 (1) crystallizes in the space group P1 is dominated by O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonding due to the presence of acetic acid and of the carb\u00adoxy\u00adlic group functionalizing the upper rim. Further stabilization is provided by offset \u03c0\u2013\u03c0 stacking inter\u00adactions between the aromatic walls of adjacent cavitands [inter\u00adcentroid distance = 3.573\u2005(1)\u2005\u00c5].The cavitand 5,11,17,23-tetra\u00admethyl-4,24:6,10:12,16:18,22-tetra\u00adkis\u00ad(methyl\u00adenedi\u00adoxy)resorcin[4]arene functionalized at the upper rim with a carb\u00adoxy\u00adlic acid group, It is therefore important to find an accurate method to measure acetic acid vapour in order to assess the environmental air quality. In the literature, only one example of the environmental monitoring of gaseous acetic acid has been reported is also shown in Fig.\u00a011 crystallizes in the space group PC, while the methyl group of the acetic acid held inside the cavity forms C\u2014H\u22ef\u03c0 inter\u00adactions with the aromatic rings of the walls , as can be seen from the C3\u2014O1\u2014C9\u2014O2 torsion angles . This is probably due to the hydrogen bonding in which the carb\u00adoxy\u00adlic acid C9D/C10D/O3D/O4D is involved with adjacent cavitands, as will be described in Section 3.1 is dominated by hydrogen bonding. A chain which propagates along the c-axis direction is formed by strong O\u2014H\u22efO inter\u00adactions involving the hydroxyl group O3D\u2014H3D from the carb\u00adoxy\u00adlic acid at the methyl\u00adene bridge and the bridging resorcinol oxygen atom O2Bi of an adjacent cavitand \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01; see Fig.\u00a04D-\u2013H7D1\u22efO1ii] and an intra\u00admolecular one with a methyl\u00adene bridge [C9A\u2014H9A1\u22efO1]. These sets of inter\u00adactions are completed by another inter\u00admolecular C\u2014H\u22efO hydrogen bond between methyl group C7C\u2014H7C2 and the carboxyl oxygen atom O4Dii. Finally, the ribbons C1A\u2013C6A aromatic rings [Fig.\u00a05While the main supra\u00admolecular contacts at play for the encapsulation of acetic acid inside the cavitand are C\u2014H\u22ef\u03c0 inter\u00adactions Table\u00a01, the crynd Fig.\u00a03. Pairs o5H11 alkyl chains at the lower rim (XIDLEC) have been used to form supra\u00admolecular complexes with di\u00admethyl\u00admethyl\u00adphospho\u00adnate, DMMP, a nerve-gas simulant bearing a P=O group has been used to form a heterodimeric capsule in a rim-to-rim fashion through the formation of four hydrogen bonds with a tetra\u00ad(3-pyrid\u00adyl)-cavitand. The previously cited cavitand A self-assembles into a one-dimensional chain (LOPKEG) or into dimeric capsules (KAHMOV) via hydrogen bonding with four 2-amino\u00adpyrimidine mol\u00adecules. Similarly, OSIYIA and OSIYOG consist of supra\u00admolecular self-assembled polymers or capsules between tetra\u00adcarb\u00adoxy\u00adlic acid functionalized cavitands and suitable N-heterocyclic linkers such as 4,4-bi\u00adpyridine and 2-amino-5-bromo-4-chloro-6-methyl\u00adpyrimidine.A resorcinarene-based cavitand in which one of the four methyl\u00adenic bridges is functionalized with a carb\u00adoxy\u00adlic acid is unique to the present day. An isomer of the title compound (XIDLIG) and its analogue with four \u2013CCavCOOH-in was carried out according to the procedure employed for the CavCOOH-out isomer spectrometer. All chemical shifts (\u03b4) are reported in p.p.m. relative to the proton resonances resulting from incomplete deuteration of the NMR solvents. 1H NMR d = 1.91 , 2.01 , 3.23 , 4.31 , 4.51 , 5.85 , 6.73 , 6.94 .The synthesis of cavitand 1 were obtained by slow evaporation of a solution prepared by dissolving 0.005\u2005mmol of the cavitand CavCOOH-in in 5\u2005ml of a 1:1 di\u00adchloro\u00admethane and acetone solution, to which 1.1\u2005\u00b5L (0.02\u2005mmol) of glacial acetic acid were added.Colourless crystals of the inclusion complex Uiso(H) set to 1.2\u20131.5Ueq(C/O), the only exception being atom H9D, which was located in a difference-Fourier map and refined freely. A DIFX instruction was employed to avoid a short H\u22efH contact between atoms H9D and H8D1. Atoms O1 and O2 were refined using the EADP command. The acetic acid guest is disordered over two positions with a refined occupancy ratio of 0.344\u2005(4):0.656\u2005(4).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019002512/su5481sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989019002512/su5481Isup2.hklStructure factors: contains datablock(s) I. DOI: 1897735CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Cervical sagittal parameters were closely related with clinical outcomes after multi-level ACDF. Our purpose was to evaluate the clinical outcomes and cervical sagittal parameters in patients with MCSM after ACDF and to identify the risk factors of poor clinical outcomes.ACDF was performed in 89 patients with MCSM. Based on average JOA recovery rate, patients were divided good-outcome group (group GO) and poor-outcome group (group PO). The cervical sagittal parameters including Cobb angle, SVA, T1S, cranial tilt and cervical tilt were measured. Multivariate logistic regression was used to identify risk factors.p\u2009<\u20090.001). \u25b3Cobb angle (p\u2009=\u20090.008) and \u25b3SVA (p\u2009=\u20090.009) showed significantly statistical differences between two groups. Longer symptom duration, lower preoperative JOA score, smaller \u25b3Cobb angle and larger \u25b3SVA were identified as risk factors of poor clinical outcomes.Fifty-four patients (60.67%) were divided into group GO, while 35 patients (39.33%) were divided into group PO. Cobb angle, SVA and T1S was corrected from preoperative average 11.80\u00b0\u2009\u00b1\u20099.63\u00b0, 23.69\u2009mm\u2009\u00b1\u200911.69\u2009mm and 24.43\u00b0\u2009\u00b1\u200911.78\u00b0 to postoperative average 15.08\u00b0\u2009\u00b1\u20099.05\u00b0, 18.79\u2009mm\u2009\u00b1\u200910.78\u2009mm and 26.92\u00b0\u2009\u00b1\u200911.94\u00b0 respectively (Multi-level ACDF is an effective surgical method to treat patients with MCSM. However, long duration of preoperative symptoms, lower preoperative JOA score, smaller \u25b3Cobb angle and larger \u25b3SVA are risk factors for poor outcomes in patients with MCSM after ACDF. Sagittal parameters should be paid attention to in surgery. Cervical spondylotic myelopathy (CSM) is caused by spinal cord compression as a result of multiple pathological changes such as disc herniation, degeneration and/or osteophyte formation at the posterior margin of the vertebral body . Multi-lAs an anterior procedure surgery, multi-level ACDF was widely used in treating MCSM . AlthougAll protocols of the study were approved by the Ethics Committee of the Third Hospital of Hebei Medical University and informed consent was obtained from all individual participants for using their imaging data and questionnaire scores.From January 2010 to December 2015, 89 patients, including 40 men and 49 women, diagnosed as MCSM according to clinical manifestations and imaging scans underwent multi-level ACDF at the Department of Spinal Surgery, the Third Hospital of Hebei Medical University, were enrolled in this retrospective study. The inclusion criteria were the following: (1) MCSM required surgical treatment for equal to or more than three levels; (2) ineffective conservative treatment for more than 3\u00a0months or acute aggravated neurological deficit; (3) complete imaging and clinical date; (4) follow-up for at least 2\u2009years. Exclusion criteria were the following: (1) history of operation involving with cervical spine; (2) combined with trauma, spinal tumours, spinal tuberculosis or infections; (3) ossification of posterior longitudinal ligament; (4) combined with severe osteoporosis; (5) combined with neurological diseases, such as vitamin B deficiency or motor neuron diseases. The average age of all patients at operation was 58.97\u2009\u00b1\u20095.79\u2009years, range from 37 to 78\u2009years. Three-level ACDF (C3-C6 in 34 cases and C4-C7 in 41 cases) was performed in 75 patients and four-level ACDF (C3-C7) in 14 patients. The patients were followed up for an average of 2.57\u2009\u00b1\u20090.78\u2009years.Japanese Orthopaedic Association (JOA) scoring system (0\u201317 scores) was used for neurological function assessment before surgery and at last follow-up visit. Neurological function recovery rate was calculated on the basis of JOA scoring system: (postoperative score-preoperative score)/(17-preoperative score)\u2009\u00d7\u2009100%. According to average JOA scores, patients were divided into two groups: good-outcome group and poor-outcome group .Cervical magnetic resonance imaging (MRI), CT (Computed Tomography) and posterior-anterior and lateral X-rays were taken preoperatively to diagnose and evaluate the disease and make surgical planning. Cervical posterior-anterior and lateral X-rays were also taken postoperatively and at each follow-up visit. Cervical sagittal parameters were measured on lateral radiographs on synapse system with patients in a neutral position. The measurement methods of cervical sagittal parameters were as follows , duration of symptoms, follow-up period, preoperative JOA scores, number of operative segments, \u25b3Cobb angle, \u25b3SVA, \u25b3T1S, \u25b3cranial tilt and \u25b3cervical tilt were collected for potential risk factors of poor clinical outcomes in patients with MCSM after ACDF. Duration of symptoms was defined as the period from appearance of primary neurological clinical symptoms to surgery.p value <\u20090.05 was considered statistically significant. Quantitative dates were first tested its normality and homogeneity of variance and according to different situations, they were tested by Student\u2019s t test or Mann\u2013Whitney U test. Qualitative date was tested by chi-square test. The potential risk factors were tested by univariate analysis and if p\u2009<\u20090.05, the factor was selected into multivariate logistic model. Then, multivariate logistic regression analysis was used to identify the risk factors of poor clinical outcomes in patients with MCSM after ACDF with adjusted odds ratios (ORs) and 95% confidence intervals (CIs).Clinical and imaging date was evaluated by SPSS program . p\u2009<\u20090.001). The average recovery rate was 61.13%\u2009\u00b1\u200921.48%. Fifty-four patients (60.67%) whose recovery rate higher than the average were divided into group GO and their average recovery rate was 75.00%\u2009\u00b1\u20099.52%, while 35 patients (39.33%) whose recovery rate lower than the average were divided into group PO and their average recovery rate was 39.73%\u2009\u00b1\u200916.62%. There was significantly statistical difference in recovery rate between two groups (p\u2009<\u20090.001).For all patients, the mean duration of symptoms was 14.64\u2009\u00b1\u20098.02\u2009months. All the operations were completed successfully Fig.\u00a0. JOA scop\u2009=\u20090.114), sex (p\u2009=\u20090.450), BMI (p\u2009=\u20090.582), follow-up time (p\u2009=\u20090.159) and operative segment (p\u2009=\u20090.768) between two groups. The JOA scores were significantly improved at last follow-up visit comparing with the preoperative scores in both groups (p\u2009<\u20090.001). No matter preoperative JOA scores (p\u2009=\u20090.014) or last follow-up JOA scores (p\u2009<\u20090.001), group GO got significantly higher scores than group PO. In addition, patients in group PO complained of syndromes for a significantly longer period before surgery than patients in group GO (p\u2009<\u20090.001).The clinical dates between group GO and group PO were showed in Table\u00a0p\u00a0<\u20090. 001) and the average \u25b3Cobb angle was 3.28\u00b0\u2009\u00b1\u20093.88\u00b0. The average last follow-up SVA (18.79\u2009mm\u2009\u00b1\u200910.78\u2009mm) was significantly lower than the preoperative (23.69\u2009mm\u2009\u00b1\u200911.69\u2009mm) (p\u00a0<\u20090.001) and the average \u25b3SVA was \u2212\u20094.90\u2009mm\u2009\u00b1\u20096.30\u2009mm. Meanwhile, significantly significant difference was also found between preoperative T1S (24.43\u00b0\u2009\u00b1\u200911.78\u00b0) and last follow-up T1S (26.92\u00b0\u2009\u00b1\u200911.94\u00b0) (p\u00a0<\u20090.001) and the average \u25b3T1S was 2.49\u00b0\u2009\u00b1\u20094.19\u00b0. Significantly statistical difference (p\u00a0<\u20090.001) was found between preoperative (5.16\u00b0\u2009\u00b1\u20096.47\u00b0) and last follow-up cranial tilt (7.52\u00b0\u2009\u00b1\u20096.27\u00b0) and the average \u25b3cranial tilt was 2.36\u00b0\u2009\u00b1\u20092.25\u00b0.There was no significantly statistical difference (p\u00a0=\u20090.132) between preoperative (17.71\u00b0\u2009\u00b1\u20096.28\u00b0) and last follow-up cervical tilt (17.46\u00b0\u2009\u00b1\u20096.54\u00b0) and the average \u25b3cervical tilt was \u2212\u20090.25\u00b0\u2009\u00b1\u20091.53\u00b0.For all patients, Cobb angle was corrected from preoperative average 11.80\u00b0\u2009\u00b1\u20099.63\u00b0 to postoperative average 15.08\u2009\u00b1\u20099.05\u00b0 (p\u00a0=\u20090.467), preoperative SVA (p\u00a0=\u20090.868) and preoperative T1S (p\u00a0=\u20090.740) between two groups. However, last follow-up Cobb angle in group GO was greater than that in group PO (p\u00a0=\u20090.025) and significantly statistical difference was also found when comparing \u25b3Cobb angle (p\u00a0=\u20090.008). Last follow-up SVA showed the opposite result than the value in group PO was greater than that in group GO (p\u00a0=\u20090.030). \u25b3SVA in group PO was also greater than that in group GO (p\u00a0=\u20090.009). There was no significantly statistical difference in both last follow-up T1S (p\u00a0=\u20090.814) and \u25b3T1S (p\u00a0=\u20090. 826) between two groups. No significantly statistical difference was found in preoperative cranial tilt (p\u00a0=\u20090.740), last follow-up cranial tilt (p\u00a0=\u20090.653), \u25b3cranial tilt (p\u00a0=\u20090.952), preoperative cervical tilt (p\u00a0=\u20090.590), last follow-up cervical tilt (p\u00a0=\u20090.585) and \u25b3cervical tilt (p\u00a0=\u20090.946) between two groups.The comparison of cervical sagittal parameters between group GO and group PO is shown in Table\u00a0p\u2009<\u20090.001), preoperative JOA score (p\u2009=\u20090.009), \u25b3Cobb angle (p\u2009=\u20090.013) and \u25b3SVA (p\u2009=\u20090.001) showed significantly statistical difference in univariate analysis and the four factors were selected into multivariate logistic model (Table\u00a0p\u2009<\u20090.001), lower preoperative JOA score , smaller \u25b3Cobb angle and larger \u25b3SVA were identified as four risk factors of poor clinical outcomes in patients with MCSM after ACDF is one of the most common diseases in orthopaedics and is one of the most harmful diseases, mostly in the elderly . It has The clinical outcomes of ACDF were limited by a variety of factors involving with preoperative condition and postoperative complications. Pumberger et al. found thCervical sagittal parameters had been proved to be important in clinical recovery of patients with CMS after cervical surgery and preoperative cervical sagittal parameters had been proved to be predictors for clinical outcomes \u201324. CervThe similar result showed in the study of Basques et al. that ACDMulti-level ACDF is an effective surgical method to treat patients with MCSM. Cervical sagittal parameters were changed after multi-level ACDF with larger Cobb angle, smaller SVA and greater T1S. However, long duration of preoperative symptoms, lower preoperative JOA score, smaller \u25b3Cobb angle and larger \u25b3SVA is risk factors for poor outcomes in patients with MCSM after ACDF. Sagittal parameters should be paid attention to in design of surgical plan for better clinical outcomes."} +{"text": "In the crystal, [010] polymeric chains are crosslinked by N\u2014H\u22efO hydrogen bonds to form sheets lying parallel to the (001) plane.The title com\u00adpound is one-dimensional coordination polymer built up of tetra\u00adgonally distorted CuN catena-poly[[[(perchlorato-\u03baO)copper(II)]-\u03bc-3-(3-carb\u00adoxy\u00adprop\u00adyl)-1,5,8,12-tetra\u00adaza-3-azonia\u00adcyclo\u00adtetra\u00addecane-\u03ba4N1,N5,N8,N12] bis\u00ad(per\u00adchlorate)], {[Cu(C13H30N5O2)(ClO4)](ClO4)2}n, (I), consists of a macrocyclic cation, one coordinated per\u00adchlorate anion and two per\u00adchlorate ions as counter-anions. The metal ion is coordinated in a tetra\u00adgonally distorted octa\u00adhedral geometry by the four secondary N atoms of the macrocyclic ligand, the mutually trans O atoms of the per\u00adchlorate anion and the carbonyl O atom of the protonated carb\u00adoxy\u00adlic acid group of a neighbouring cation. The average equatorial Cu\u2014N bond lengths [2.01\u2005(6)\u2005\u00c5] are significantly shorter than the axial Cu\u2014O bond lengths [2.379\u2005(8)\u2005\u00c5 for carboxyl\u00adate and average 2.62\u2005(7)\u2005\u00c5 for disordered per\u00adchlorate]. The coordinated macrocyclic ligand in (I) adopts the most energetically favourable trans-III conformation with an equatorial orientation of the substituent at the protonated distal 3-position N atom in a six-membered chelate ring. The coordination of the carb\u00adoxy\u00adlic acid group of the cation to a neighbouring com\u00adplex unit results in the formation of infinite chains running along the b-axis direction, which are cross\u00adlinked by N\u2014H\u22efO hydrogen bonds between the secondary amine groups of the macrocycle and O atoms of the per\u00adchlorate counter-anions to form sheets lying parallel to the (001) plane. Additionally, the extended structure of (I) is consolidated by numerous intra- and interchain C\u2014H\u22efO contacts.The asymmetric unit of the title com\u00adpound, N3,N10-disubstituted 1,3,5,8,10,12-hexa\u00adaza\u00adcyclo\u00adtetra\u00addecane (di\u00adaza\u00adcyclam) and, to a lesser extent, N3-substituted 1,3,5,8,12-penta\u00adaza\u00adcyclo\u00adtetra\u00addecane (aza\u00adcyclam) are popular building units for the assembly of metal\u2013organic frameworks (MOFs), demonstrating many promising applications or 3,7-di\u00adaza\u00adnonane-1,9-di\u00adamine com\u00adplexes, respectively, with formaldehyde and primary amines catena-poly[[[(perchlorato-\u03baO)copper(II)]-\u03bc-3-(3-carb\u00adoxy\u00adprop\u00adyl)-1,5,8,12-tetra\u00adaza-3-azonia\u00adcyclo\u00adtetra\u00addecane-\u03ba4N1,N5,N8,N12] bis\u00ad(per\u00adchlorate)], {[Cu(H2L)(ClO4)](ClO4)2}n, which is the first example of aza\u00adcyclam ligand with a carb\u00adoxy\u00adlic acid group.Herein, we describe the synthesis and the crystal structure of the title CuII ion in the com\u00adplex cation in (I)II ion is displaced by 0.075\u2005\u00c5 from the mean plane of the N4 donor atoms (r.m.s. deviation = 0.005\u2005\u00c5) towards the O2 atom of the carboxyl\u00adate group. The equatorial Cu\u2014N bond lengths are significantly shorter than the axial Cu\u2014O bond lengths conformation \u00b0] and the six-membered chelate rings in chair [average bite angle = 93.6\u2005(2)\u00b0] conformations. The methyl\u00adene group of the substituent at the noncoordinated N3 atom in the six-membered chelate ring is oriented equatorially. Such an arrangement of the substituent, in contrast to an axial orientation, is relatively uncommon and only a few examples of such CuII com\u00adplexes with aza- and di\u00adaza\u00adcyclam ligands have been described so far + group in (I)sp3-hybridized N atom (ca 327\u00b0), thus indicating their partial sp2 character -substituted di\u00adaza\u00adcyclam polymeric com\u00adplex and 1.198\u2005(13)\u2005\u00c5 for C13\u2014O1 and C13\u2014O2, respectively], thus confirming its protonated form and the lack of delocalization. Inter\u00adestingly, it is coordinated to the Cu4 anion is com\u00adpletely disordered over two positions with site occupancies of 50% and is weakly coordinated to the metal ion (Table\u00a014) and 78% (Cl3O4). Because of the low partial population, the minor com\u00adponents of these per\u00adchlorate anions were not considered further in the analysis of the hydrogen-bonding network.Three disordered per\u00adchlorate anions in the title com\u00adpound counterbalance the charge of the com\u00adplex cations. The Cl1On Table\u00a01. Two remb-axis direction ,O12(Cl3)], so that each per\u00adchlorate anion is fixed in a chain in a ditopic manner ], as well as between secondary amine groups of the macrocycle and an O atom of the carb\u00adoxy\u00adlic acid group as the proton acceptor [N2\u2014H2(N4\u2014H4)\u22efO1]. Hydrogen bonding of the secondary amine groups of the macrocycle and the O atoms of per\u00adchlorate anions not involved in above-mentioned intra\u00adchain inter\u00adactions [N1\u2014H1\u22efO7(Cl2)(x\u00a0\u2212\u00a0y\u00a0+\u00a0z) and N5\u2014H5\u22efO14(Cl3)(x\u00a0\u2212\u00a0y\u00a0+\u00a0z)] results in the formation of sheets lying parallel to the (001) plane and many of them were investigated as building blocks for the construction of MOFs by using additional carboxyl\u00adate or metalocyanide linkers. At the same time, there are only two examples demonstrating self-polymerization , namely, those with di\u00adaza\u00adcyclam ligands containing 2-propio\u00adnitrile ](ClO4)2 , was prepared according to a published method ](ClO4)2 , 4-amino\u00adbutanoic acid and 30% aqueous formaldehyde in methanol (40\u2005ml) was refluxed for 24\u2005h. After cooling and filtration, the solution was kept in a refrigerator overnight. The violet crystalline precipitate was filtered off, washed with methanol (5\u2005ml) and recrystallized from a 1:1 (v/v) water\u2013ethanol solvent mixture (10\u2005ml) containing 0.5\u2005M perchloric acid . Analysis calculated (%) for C13H30Cl3CuN5O14: C 24.01, H 4.65, N 10.76; found: C 24.17, H 4.51, N 10.92. Violet blocks of (I)All chemicals and solvents used in this work were purchased from Sigma\u2013Aldrich and were used without further purification. The starting CuSafety note: per\u00adchlorate salts of metal com\u00adplexes are potentially explosive and should be handled with care.Uiso(H) values of 1.2 or 1.5Ueq of the parent atoms. The crystal of (I)Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901901377X/hb7857sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901901377X/hb7857Isup2.hklStructure factors: contains datablock(s) I. DOI: 1958285, 1958285CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "H-imidazol-3-ium) hepta\u00admolybdate 2-methyl-1H-imidazole disolvate dihydrate,(C4H7N2)6[Mo7O24]\u00b72C4H6N2\u00b72H2O, [Mo7O24]6\u2212 hepta\u00admolybdate anions, 2-methyl\u00adimidazolium cations, neutral 2-methyl\u00adimidazole mol\u00adecules and water mol\u00adecules are linked through hydrogen bonds into a three-dimensional network.In the crystal structure of hexa\u00adkis\u00ad(2-methyl-1 H-imidazol-3-ium) hepta\u00admolybdate 2-methyl-1H-imidazole disolvate dihydrate, (C4H7N2)6[Mo7O24]\u00b72C4H6N2\u00b72H2O, was prepared from 2-methyl\u00adimidazole and ammonium hepta\u00admolybdate tetra\u00adhydrate in acid solution. The [Mo7O24]6\u2212 hepta\u00admolybdate cluster anion is accompanied by six protonated (C4H7N2)+ 2-methyl\u00adimidazolium cations, two neutral C4H6N2 2-methyl\u00adimidazole mol\u00adecules and two water mol\u00adecules of crystallization. The cluster consists of seven distorted MoO6 octa\u00adhedra sharing edges or vertices. In the crystal, the components are linked by N\u2014H\u22efN, N\u2014H\u22efO, O\u2014H\u22efO, N\u2014H\u22ef and O\u2014H\u22ef hydrogen bonds, generating a three-dimensional network. Weak C\u2014H\u22efO inter\u00adactions consolidate the packing.The title compound, hexa\u00adkis\u00ad(2-methyl-1 M = V, Nb, Ta, Mo, W, \u22ef) and oxygen atoms with a structural and compositional diversity that lead to numerous applications because of their electrochemical, optical, catalytic and photochromic properties as well as their anti\u00adviral and anti\u00adtumor activities are clusters of transition metals , \u03bc2-O (oxygen atoms bridging two molybdenum atoms), \u03bc3-O (oxygen atoms bridging three molybdenum atoms) and \u03bc4-O (oxygen atoms bridging four molybdenum atoms). All of the Mo atoms are bound to two terminal oxygen atoms except for Mo7, which is located in the \u2018core\u2019 of the cluster. The geometrical data for the cluster in (I)et al., 1992et al., 2008t, 1.754\u2005(2)\u20142.453\u2005(2)\u2005\u00c5 for \u03bc2-O, 1.8945\u2005(19)\u20132.3057\u2005(19)\u2005\u00c5 for \u03bc3-O and 2.1329\u2005(19)\u20132.3011\u2005(18)\u2005\u00c5 for \u03bc4-O. The variations of Mo\u2014O bond lengths and O\u2014Mo\u2014O angles indicate that all seven octa\u00adhedra (MoO6) within the cluster are highly distorted. As in the compound (H3dien)2[Mo7O24]\u00b74H2O . As well as the [Mo7O24]6\u2212 anion, six (C4H7N2)+ cations, two neutral C4H6N2 mol\u00adecules and two water mol\u00adecules of crystallization are present in the asymmetric unit + cations and/or neutral 2-methyl\u00adimidazole mol\u00adecules + cations to neutral mol\u00adecules. The packing is consolidated by weak C\u2014H\u22efO links , 2-methyl\u00adimidazole and ammonium hepta\u00admolybdate tetra\u00adhydrate in a ratio of 1:2:1/12 were dissolved in water (60\u2005ml). The solution was stirred for one\u2005h and evaporated in the oven at 333\u2005K to yield a whitish precipitate. The precipitate was recrystallized from methanol solution: after two weeks at room temperature, colourless prisms of (I)supporting information. The absorption bands at 3400 and 3395\u2005cm\u22121 corres\u00adpond to \u03bd(O\u2014H) stretches and indicate the presence of water mol\u00adecules and those at 1621 and 1564\u2005cm\u22121 to the deformation vibrations \u03b4(O\u2014H). The bands centered at 3132 and 1431\u2005cm\u22121 with shoulders are respectively attributed to the stretching and deformation vibrations of the N\u2014H bonds of the protonated and/or non-protonated entities of 2-methyl\u00adimidazole deformation vibration stretching vibrations while the bands between 838 and 650\u2005cm\u22121 are typical for the vibrations of \u03bd(Mo\u2014O\u2014Mo) and \u03bd[Mo\u2014(\u03bc-O)] groupings Uiso (H) = 1.2Ueq or 1.5Ueq(C-meth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019008454/hb7831sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019008454/hb7831Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019008454/hb7831sup3.tifThe IR spectrum of compound (I). DOI: 1922927CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The structure of the pseudopolymorph of 3-(tri\u00adphenyl\u00adphospho\u00adranyl\u00adidene)-2,5-di\u00adhydro\u00adfuran-2,5-dione with a THF solvent mol\u00adecule is described. The compound has a hydrogen-bonded layer structure, and displays C\u2014H\u22efO hydrogen bonds connecting mol\u00adecules of the di\u00adhydro\u00adfuran-2,5-dione derivative into chains. viz. C22H17O3P\u00b7C4H8O. The succinic anhydride ring is approximately planar (r.m.s. deviation = 0.032\u2005\u00c5). The tetra\u00adhydro\u00adfuran mol\u00adecule is disordered over two orientations about a pseudo-twofold axis with refined occupancy ratio 0.718\u2005(4):0.282\u2005(4). In the crystal, C\u2014H\u22efO hydrogen bonds link mol\u00adecules of the di\u00adhydro\u00adfuran-2,5-dione derivative into chains parallel to the b axis and arranged into layers stacked along [100] alternating with hydrogen-bonded tetra\u00adhydro\u00adfuran layers.The title pseudo-polymorph of 3-(tri\u00adphenyl\u00adphospho\u00adranyl\u00adidene)-2,5-di\u00adhydro\u00adfuran-2,5-dione crystallizes with a tetra\u00adhydro\u00adfuran solvent mol\u00adecule, The distribution histogram of the P=C distance [with a mean value of 1.729\u2005\u00c5 and a standard deviation of 0.030\u2005\u00c5] is shown in Fig.\u00a03et al., 19932Cl2 solvate : 1.78 , 3.14 , 3.67 , 7.44\u20137.72 . 31P{1H} NMR : +13.6 (s).To a stirred solution maleic anhydride in tetra\u00adhydro\u00adfuran THF (5\u2005mL) was added tri\u00adphenyl\u00adphosphine at room temperature. The reaction mixture was stirred at room temperature for 24\u2005h, then the solution was filtered and concentrated under reduced pressure. The reaction mixture was allowed to cool in the freezer and yellowish crystals precipitated. The crystals were separated from solvent and dried to give 0.56\u2005g (90%) of the title compound. Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018011775/rz5234sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018011775/rz5234Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018011775/rz5234Isup3.cdxSupporting information file. DOI: 1862873CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Both mol\u00adecules feature an intra\u00admolecular N\u2014H\u22efN hydrogen bond. In the crystal, weak aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid\u2013centroid separation = 3.830\u2005(2)\u2005\u00c5] link the mol\u00adecules into dimers.In each of the two independent mol\u00adecules in the asymmetric unit of the title compound, [CdI For A, the Cd1\u2014I1 and Cd1\u2014I2 bond lengths are 2.7509\u2005(4) and 2.6917\u2005(4)\u2005\u00c5, respectively, and the Cd1\u2014N2 bond length is 2.342\u2005(3)\u2005\u00c5. For B, the Cd2\u2014I3 and Cd2\u2014I4 bond lengths are 2.7530\u2005(4) and 2.6732\u2005(4)\u2005\u00c5, respectively, and the Cd2\u2014N6 bond length is 2.344\u2005(3)\u2005\u00c5. Both mol\u00adecules feature an intra\u00admolecular N\u2014H\u22efN hydrogen bond with the pyridine-ring N atom as the acceptor \u00b0 for r Table\u00a02.A + B associations by aromatic \u03c0\u2013\u03c0 stacking inter\u00adactions y] Fig.\u00a03.CrystalExplorer3.1 , H\u22efH (29.5%), C\u22efH / H\u22efC (13.3%), H\u22efO / O\u22efH (5.6%) and C\u22efI / I\u22efC (4.9%) are shown in Figs.\u00a04b\u2013f, respectively. The full list of percentage surface contributions in given in Table\u00a03The Hirshfeld surface analysis -4-hy\u00addroxy\u00adbenzyl\u00adidene]-pyridine-4-carbohydrazide-\u03baN1}di\u00adiodido\u00adcadmium methanol disolvate ethyl\u00adidene]picolinohydrazide-\u03ba2N\u2032,O}cadmium methyl\u00adidene]pyridine-2-carbohydrazide-\u03ba2N\u2032,O}cadmium) cadmium -4-(2-hy\u00addroxy\u00adbenzyl\u00adidene\u00adamino)\u00adbenzoato-\u03ba2O,O\u2032]cadmium methyl\u00adidene]picolinohydrazide-\u03ba2N\u2032,O}cadmium N\u2032-[(E)-(pyridin-2-yl)methyl\u00adidene]pyridine-2-carbohydrazide ligand was synthesized according to the literature method = 1.2Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989019008831/hb7823sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019008831/hb7823Isup2.hklStructure factors: contains datablock(s) I. DOI: 1935658CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title Schiff base compound, consisting of a cyclo\u00adhexane and a 2-hy\u00addroxy-3-methyl\u00adbenzyl\u00adidene ring bridged by a hydrazinecarbo\u00adthio\u00adamine moiety, crystallizes with two independent mol\u00adecules in the asymmetric unit. In the crystal, the mol\u00adecules are linked by N\u2014H\u22efS hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, forming ribbons along the [010] direction. 15H21N3OS, comprises of two crystallographically independent mol\u00adecules (A and B). Each mol\u00adecule consists of a cyclo\u00adhexane ring and a 2-hy\u00addroxy-3-methyl\u00adbenzyl\u00adidene ring bridged by a hydrazinecarbo\u00adthio\u00adamine unit. Both mol\u00adecules exhibit an E configuration with respect to the azomethine C=N bond. There is an intra\u00admolecular O\u2014H\u22efN hydrogen bond in each mol\u00adecule forming an S(6) ring motif. The cyclo\u00adhexane ring in each mol\u00adecule has a chair conformation. The benzene ring is inclined to the mean plane of the cyclo\u00adhexane ring by 47.75\u2005(9)\u00b0 in mol\u00adecule A and 66.99\u2005(9)\u00b0 in mol\u00adecule B. The mean plane of the cyclo\u00adhexane ring is inclined to the mean plane of the thio\u00adurea moiety [N\u2014C(=S)\u2014N] by 55.69\u2005(9) and 58.50\u2005(8)\u00b0 in mol\u00adecules A and B, respectively. In the crystal, the A and B mol\u00adecules are linked by N\u2014H\u22efS hydrogen bonds, forming \u2018dimers\u2019. The A mol\u00adecules are further linked by a C\u2014H\u22ef\u03c0 inter\u00adaction, hence linking the A\u2013B units to form ribbons propagating along the b-axis direction. The conformation of a number of related cyclo\u00adhexa\u00adnehydrazinecarbo\u00adthio\u00adamides are compared to that of the title compound.The asymmetric unit of the title compound, C The benzene ring is inclined to the mean plane of the thio\u00adurea moiety by 10.95\u2005(8)\u00b0 in mol\u00adecule A and 9.80\u2005(8)\u00b0 in mol\u00adecule B.The cyclo\u00adhexane ring (C9\u2013C14) in each mol\u00adecule has a chair conformation. The mean plane of the four central C atoms (C10/C11/C13/C14) is inclined to the mean plane of the thio\u00adurea moiety [N2\u2014C8(=S1)\u2014N3] by 54.83\u2005(11) and 55.64\u2005(10)\u00b0 in mol\u00adecules i.e. \u03c41 (C1\u2014C6\u2014C7\u2014N1), \u03c42 (C7\u2014N1\u2014N2\u2014C8), \u03c43 (N1\u2014N2\u2014C8\u2014N3), \u03c44 (N2\u2014C8\u2014N3\u2014C9) and \u03c45 (C8\u2014N3\u2014C9\u2014C10), as illustrated in Fig.\u00a03\u03c41 between the benzyl\u00adidine ring and the azomethine double bond for both mol\u00adecules are approximately 0\u00b0 [3.0\u2005(2)\u00b0 in mol\u00adecule A and 1.9\u2005(2)\u00b0 in mol\u00adecule B], signifying the coplanarity between benzyl\u00adidine ring and the azomethine double bond (C7=N1). In mol\u00adecule B, the azomethine double bond is close to planar with the hydrazine moiety [\u03c42 = 177.23\u2005(14)\u00b0], whereas \u03c42 in mol\u00adecule A is slightly twisted [\u03c42 = 171.68\u2005(14)\u00b0]. In both mol\u00adecules, the torsion angle between the hydrazine moiety and the carbo\u00adthio group are also slight twisted with \u03c43 values in mol\u00adecules A and B of 7.4\u2005(2) and \u221210.2\u2005(2)\u00b0, respectively. Similarly to \u03c41, the carbo\u00adthio group is almost coplanar with the thio\u00adamide group for both mol\u00adecules, as implied by torsion angle \u03c44 [178.07\u2005(14)\u00b0 in mol\u00adecule A and 175.59\u2005(14)\u00b0 in mol\u00adecule B], which are approximately 180\u00b0. The thio\u00adamide group and the cyclo\u00adhexane ring are almost perpendicular to each other with \u03c45 torsion angles of 85.3\u2005(2) and \u221281.6\u2005(2)\u00b0 in mol\u00adecules A and B, respectively. This may arise from the steric repulsion between the cyclo\u00adhexane ring and adjacent sulfur atom.The unique mol\u00adecular conformations of the two mol\u00adecules can be characterized by five torsion angles, A and B mol\u00adecules are connected into \u2018dimers\u2019 with an via N2A\u2014H1N2\u22efS1Bi and N2B\u2014H2N2\u22efS1Ai hydrogen bonds -2-benzyl\u00adidene-N-cyclo\u00adhexyl\u00adhydrazine-1-carbo\u00adthio\u00adamide as the reference moiety resulted in nine structures containing a cyclo\u00adhexyl\u00adhydrazinecarbo\u00adthio\u00adamide moiety with different substituents (R). The different substituents (R) together with the torsion angles of the hydrazinecarbo\u00adthio\u00adamide connecting bridge are compiled in Table\u00a02cf. Fig.\u00a03\u03c42, \u03c43 and \u03c44 are in, respectively, anti-periplanar (153.5 to 179.3\u00b0), syn-periplanar (0.8 to 14.7\u00b0) and anti-periplanar (from 171.8 to 180.0\u00b0) conformations. The attached cyclo\u00adhexane ring is always close to perpendicular to the thio\u00adamide group and with a syn/anti-clinal (\u03c45 = 78.3 to 94.5\u00b0) conformation. Furthermore, torsion angle \u03c41 for most of these structures exists in a syn-periplanar conformation, ranging from 0 to 25.8\u00b0, but there is one outlier conformation. The cyclo\u00adhexyl\u00adhydrazinecarbo\u00adthio\u00adamide moiety of this structure is substituted with an anthracen-9-yl\u00admethyl\u00adene ring system.A search of the Cambridge Structural Database in 20\u2005ml methanol was added dropwise with stirring to the aldehyde solution. The resulting colourless solution was refluxed for 4\u2005h with stirring. A colourless precipitate was obtained on evaporation of the solvent. The crude product was washed with n-hexane (5\u2005ml). The recovered product was dissolved in aceto\u00adnitrile and purified by recrystallization. Colourless block-like crystals suitable for X-ray diffraction analysis were obtained on slow evaporation of the aceto\u00adnitrile solvent .2-Hy\u00addroxy-3-methyl\u00adbenzaldehyde was dissolved in 20\u2005ml of methanol. Glacial acetic acid (0.20\u2005ml) was added and the mixture was refluxed for 30\u2005min. A solution of Spectroscopic and analytical data: 1H NMR : \u03b4 11.27 , \u03b4 9.51 , \u03b4 8.34 , \u03b4 8.05 , \u03b4 7.39\u20136.81 , \u03b4 2.20 , \u03b4 1.87\u20131.14 ppm. 13C NMR : \u03b4 175.79 (C=S), \u03b4 154.29 (C=N), \u03b4 143.76-119.17 (C-aromatic), \u03b4 15.93 (CH3), \u03b4 52.87\u201324.90 (C-cyclo\u00adhex\u00adyl) ppm. IR : 3364 (NH), 3148 (OH), 2989(CH3), 2931 and 2854 , 1620 (C=N), 1540 , 1268 (C=S), 1218 , 1122 (C\u2014O). 1075 (C\u2014N). Elemental analysis calculated for C15H21N3OS (Mr = 291.41\u2005g\u2005mol\u22121); C, 61.77; H, 7.21; N, 14.42%; found: C, 61.81; H, 7.19; N, 14.42%.Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019008946/su5501sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989019008946/su5501Isup2.hklStructure factors: contains datablock(s) I. DOI: 1480651CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the mol\u00adecular packing, methyl\u00adene-C\u2014H\u22efO(ring carbon\u00adyl) and N(pyridazin\u00adyl) inter\u00adactions result in the formation of a supra\u00admolecular tape along the 16H18N2O3, is constructed about a central oxopyridazinyl ring (r.m.s. deviation = 0.0047\u2005\u00c5), which is connected to an ethyl\u00adacetate group at the N atom closest to the carbonyl group, and benzyl and methyl groups second furthest and furthest from the carbonyl group, respectively. An approximately orthogonal relationship exists between the oxopyridazinyl ring and the best plane through the ethyl\u00adacetate group [dihedral angle = 77.48\u2005(3)\u00b0]; the latter lies to one side of the central plane [the Nr\u2014Nr\u2014Cm\u2014Cc torsion angle being 104.34\u2005(9)\u00b0]. In the crystal, both H atoms of the N-bound methyl\u00adene group form methyl\u00adene-C\u2014H\u22efO(ring carbon\u00adyl) or N(pyridazin\u00adyl) inter\u00adactions, resulting in the formation of a supra\u00admolecular tape along the a-axis direction. The tapes are assembled into a three-dimensional architecture by methyl- and phenyl-C\u2014H\u22efO(ring carbon\u00adyl) and phenyl-C\u2014H\u22efO(ester carbon\u00adyl) inter\u00adactions. The analysis of the calculated Hirshfeld surface indicates the dominance of H\u22efH contacts to the overall surface (i.e. 52.2%). Reflecting other identified points of contact between mol\u00adecules noted above, O\u22efH/H\u22efO (23.3%), C\u22efH/H\u22efC (14.7%) and N\u22efH/H\u22efN (6.6%) contacts also make significant contributions to the surface.The title compound, C H)-ones are pyridazine derivatives, being constructed about a six-membered ring which contains two adjacent nitro\u00adgen atoms, at positions one and two, and with a carbonyl group at position three. The inter\u00adest in these nitro\u00adgen-rich heterocyclic derivatives arises from the fact that they exhibit a number of promising pharmacological and biological activities. These include anti-oxidant , the crystal and mol\u00adecular structures of the the title pyridazin-3(2H)-one derivative, (I)Pyridazin-3(2The mol\u00adecular structure of (I)2 and eight-membered {\u22efNNCH}2 synthons, respectively. The result is the formation of a supra\u00admolecular tape orientated along the a-axis direction, Fig.\u00a02a). Globally, the tapes assemble into layers in the ab plane and these stack along the c-axis direction as shown in Fig.\u00a02b). Weak inter\u00adactions contributing to the formation of the layers include methyl-C16\u2014H\u22efO1(ring carbon\u00adyl) contacts s Table\u00a02. Betweenet al., 2019dnorm in Fig.\u00a03a), the C\u2014H\u22efN contact involving the methyl\u00adene-H13A and pyridazinyl-N2 atoms are represented as bright-red spots on the surface. The diminutive red spots appearing near the methyl\u00adene-H13B and carbonyl-O1 atoms indicate the weak C\u2014H\u22efO contact, Fig.\u00a03a) and (b). The intense blue and red regions corresponding to positive and negative electrostatic potentials on the Hirshfeld surfaces mapped over electrostatic potential in Fig.\u00a04dnorm-mapped Hirshfeld surfaces in Fig.\u00a03dnorm mapped Hirshfeld surface illustrating weak inter\u00admolecular inter\u00adactions are shown in the views of Fig.\u00a05The Hirshfeld surfaces calculated for (I)a), and those delineated into H\u22efH, O\u22efH/H\u22efO, N\u22efH/H\u22efN and C\u22efH/H\u22efC and C\u22efC contacts \u2013(f); the percentage contribution from different inter\u00adatomic contacts to the Hirshfeld surfaces of (I)b), having the greatest contribution, i.e. 52.2%, to the Hirshfeld surface, a pair of beak-shaped tips at de + di \u223c2.3\u2005\u00c5 reflect the short inter\u00adatomic contact between the methyl-H5C and H16C atoms, Table\u00a02c) demonstrates two pairs of adjoining short tips at de + di \u223c2.5 and 2.6\u2005\u00c5, together with the green aligned points in the central region, which are indicative of weak C\u2014H\u22efO contacts present in the crystal. The pair of long spikes at de + di \u223c2.5\u2005\u00c5 in the fingerprint plot delineated into N\u22efH/H\u22efN contacts of Fig.\u00a06d), are the result of a potential C\u2014H\u22efN inter\u00adaction involving the methyl\u00adene-C13\u2014H13A and pyridazinyl-N2 atoms. The short inter\u00adatomic C\u22efH/H\u22efC contacts as summarized in Table\u00a02de + di \u223c2.7 and 2.8\u2005\u00c5, respectively in Fig.\u00a06e). The presence of a weak \u03c0\u2013\u03c0 contact between the oxopyridazinyl and phenyl rings is reflected in the thick arrow-like tip at de + di \u223c3.4\u2005\u00c5 in the fingerprint plot delineated into C\u22efC contacts of Fig.\u00a06f), specifically the short inter\u00adatomic C2\u22efC9 contact, Table\u00a02i.e. 2.3%, contribution from C\u22efN/N\u22efC contacts to the Hirshfeld surface.The overall two-dimensional fingerprint plot, Fig.\u00a06et al., 2007The most closely related structure to (I)H)-one precursor. To this pyridazine (0.05\u2005mol) was added potassium carbonate (0.1\u2005mmol), tetra\u00adbutyl\u00adammonium bromide (0.01\u2005mmol) and 2-ethyl bromo\u00adacetate (0.1\u2005mol) in di\u00admethyl\u00adformamide (20\u2005ml). The mixture was stirred for 24\u2005h at room temperature. At the end of the reaction, the solution was filtered and the solvent evaporated under reduced pressure. The residue was washed with water and methyl\u00adenechloride. The solvent was removed and colourless blocks of (I)A mixture of 3-benzyl\u00adidene-4-oxo\u00adpenta\u00adnoic acid (0.05\u2005mol) and hydrazine hydrate (0.1\u2005mol) in ethanol (100\u2005ml) was refluxed for 2\u2005h. The precipitate formed was filtered off and recrystallized from acetone to obtain the 5-benzyl-6-methyl\u00adpyridazin-3(2Uiso(H) set to 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S205698901900241X/hb7802sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S205698901900241X/hb7802Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901900241X/hb7802Isup3.cmlSupporting information file. DOI: 1897511CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Bnz\u22efNPrpnit hydrogen bonds, are linked into stepped ribbons extending parallel to [110] by C\u2014HPrpnit\u22efOThz (Thz = thia\u00adzine) hydrogen bonds. The ribbons are joined into pairs by inversion-related C=O\u22efCl inter\u00adactions.In the title compound, the di\u00adhydro\u00adbenzo\u00adthia\u00adzine moiety is folded about the S1\u22efN1 axis. In the crystal, inversion dimers, generated by C\u2014H 18H12Cl2N2OS, consists of a di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit linked by a \u2013CH group to a 2,4-di\u00adchloro\u00adphenyl substituent, and to a propane\u00adnitrile unit is folded along the S\u22efN axis and adopts a flattened-boat conformation. The propane\u00adnitrile moiety is nearly perpendicular to the mean plane of the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit. In the crystal, C\u2014HBnz\u22efNPrpnit and C\u2014HPrpnit\u22efOThz hydrogen bonds link the mol\u00adecules into inversion dimers, enclosing R22(16) and R22(12) ring motifs, which are linked into stepped ribbons extending along [110]. The ribbons are linked in pairs by complementary C=O\u22efCl inter\u00adactions. \u03c0\u2013\u03c0 contacts between the benzene and phenyl rings, [centroid\u2013centroid distance = 3.974\u2005(1)\u2005\u00c5] may further stabilize the structure. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (23.4%), H\u22efCl/Cl\u22efH (19.5%), H\u22efC/C\u22efH (13.5%), H\u22efN/N\u22efH (13.3%), C\u22efC (10.4%) and H\u22efO/O\u22efH (5.1%) inter\u00adactions. Hydrogen bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Computational chemistry calculations indicate that the two independent C\u2014HBnz\u22efNPrpnit and C\u2014HPrpnit\u22efOThz hydrogen bonds in the crystal impart about the same energy (ca 43\u2005kJ\u2005mol\u22121). Density functional theory (DFT) optimized structures at the B3LYP/6\u2013311\u2005G level are compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.The title compound, C These mol\u00adecules exhibit a wide range of biological applications indicating that the 1,4-benzo\u00adthia\u00adzine moiety is a potentially useful template in medicinal chemistry research and has therapeutic applications as anti-inflammatory (Trapani A (C1\u2013C6), is oriented at a dihedral angle of 1.89\u2005(6)\u00b0 with respect to the phenyl ring, C (C10\u2013C15). A puckering analysis of the heterocyclic ring B (S1/N1/C1/C6\u2013C8) of the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit gave the parameters QT = 0.1983\u2005(15)\u2005\u00c5, q2 = 0.1957\u2005(17)\u2005\u00c5, q3 = 0.0323\u2005(19)\u2005\u00c5, \u03c6 = 354.6\u2005(6)\u00b0 and \u03b8 = 80.8\u2005(5)\u00b0, indicating it adopts a flattened-boat conformation. The propane\u00adnitrile moiety is essentially perpendicular to the di\u00adhydro\u00adbenzo\u00adthia\u00adzine unit, as indicated by the C7\u2014N1\u2014C16\u2014C17 torsion angle of 88.6\u2005(2)\u00b0. In heterocyclic ring B, the C1\u2014S1\u2014C8 [103.69\u2005(9)\u00b0], S1\u2014C8\u2014C7 [121.12\u2005(14)\u00b0], C8\u2014C7\u2014N1 [120.59\u2005(17)\u00b0], C7\u2014N1\u2014C6 [126.27\u2005(16)\u00b0], C6\u2014C1\u2014S1 [123.84\u2005(15)\u00b0] and N1\u2014C6\u2014C1 [121.46\u2005(17)\u00b0] bond angles are enlarged when compared with the corresponding values in the closely related compounds, (2Z)-2-(4-chloro\u00adbenzyl\u00adidene)-4-[2-(2-oxooxazoliden-3-yl) eth\u00adyl]-3,4-di\u00adhydro-2H-1,4-benzo\u00adthia\u00adzin-3-one, (II), -2-[(4-fluoro\u00adbenzyl\u00adidene]-4-(prop-2-yn-1-yl)-3,4 -di\u00adhydro-2H-1,4-benzo\u00adthia\u00adzin-3-one, (III), -4-[2-eth\u00adyl]-2(phenyl\u00admethyl\u00adidene)-3,4-di\u00adhydro-2H-1,4-benzo\u00adthia\u00adzin-3-one, (IV), -2-[methyl\u00adidene]-4-[2-eth\u00adyl]3,4-di\u00adhydro-2H-1,4-benzo\u00adthia\u00adzin-3-one, (V), Bnz\u22efNPrpnit (Bnz = benzene and Prpnit = propane\u00adnitrile) hydrogen bonds (Table\u00a01Prpnit\u22efOThz (Thz = thia\u00adzine) hydrogen bonds Cg1) and 2,4-dichlorophenyl rings [Cg1\u22efCg3 = 3.974\u2005(1)\u2005\u00c5] may further stabilize the structure.In the crystal, inversion dimers are formed by C\u2014Hs Table\u00a01, enclosis Table\u00a01, enclosi)\u00b0 Fig.\u00a03. The conCrystalExplorer17.5 analysis have a nearly symmetrical distribution of points, Fig.\u00a07c, with the thin edges at de + di = 2.82\u2005\u00c5. In the absence of C\u2014H\u22ef\u03c0 inter\u00adactions, the wings in the fingerprint plot delineated into H\u22efC/C\u22efH contacts (13.5%) also have a nearly symmetrical distribution of points, Fig.\u00a07d, with the thick edges at de + di \u223c2.90\u2005\u00c5. The wings in the fingerprint plot delineated into H\u22efN/N\u22efH contacts have as pair of spikes with the tips at de + di = 2.30\u2005\u00c5. The C\u22efC contacts have an arrow-shaped distribution of points with the tip at de = di \u223c1.78\u2005\u00c5. The H\u22efO/O\u22efH and C\u22efCl/Cl\u22efC contacts and C\u22efS/S\u22efC contacts are seen as pairs of wide spikes with the tips at de + di \u223c3.30 and 3.48\u2005\u00c5, respectively.The overall two-dimensional fingerprint plot, Fig.\u00a07H Table\u00a02, contribs Table\u00a02 are viewdnorm plotted onto the surface are shown for the H\u22efH, H\u22efCl/Cl\u22efH, H\u22efC/C\u22efH, H \u22ef N/N\u22efH, C\u22efC and H\u22efO/O\u22efH inter\u00adactions in Fig.\u00a08a\u2013f, respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efCl/Cl\u22efH, H \u22ef C/C\u22efH and H\u22efN/N\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing is the sum of electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies were calculated to be \u221213.0 (Eele), \u22121.8 (Epol), \u221268.0 (Edis), 48.3 (Erep) and \u221244.4 (Etot) for the C\u2014HBnz\u22efNPrpnit hydrogen-bonding inter\u00adaction and \u221237.3 (Eele), \u22129.3 (Epol), \u221219.0 (Edis), 33.7 (Erep) and \u221242.0 (Etot) for C\u2014HPrpnit\u22efOThz.The inter\u00admolecular inter\u00adaction energies were calculated using the CE\u2013B3LYP/6\u201331G energy model available in via density functional theory (DFT) using standard B3LYP functional and 6\u2013311\u2005G basis-set calculations -2-[methyl\u00adidene]-3-oxo-3,4-di\u00adhydro-2H-1,4-benzo\u00adthia\u00adzin-4-yl]propane\u00adnitrile ring. The energy band gap [\u0394E = ELUMO\u00a0\u2212\u00a0EHOMO] of the mol\u00adecule is about 6.1979\u2005eV, and the frontier mol\u00adecular orbital energies, EHOMO and ELUMO are \u22127.1543 and \u22120.9564\u2005eV, respectively.The optimized structure of the title compound in the gas phase was generated theoretically et al., 2016R1 = Ph, R2 = C), gave 14 hits. With R1 = Ph and R2 = CH2C\u2261CH IIa to 36\u00b0 (IIf). The other three have the benzo\u00adthia\u00adzine unit nearly planar with a corresponding dihedral angle of ca 3\u20134\u00b0.A search in the Cambridge Structural Database (Groom Z)-2--2H-1,4-benzo\u00adthia\u00adzin-3(4H)-one (1.8\u2005mmol), potassium carbonate (2.0\u2005mmol) and tetra n-butyl ammonium bromide (0.15\u2005mmol) in DMF (20\u2005ml). Stirring was continued at room temperature for 12 h. The salts were removed by filtration and the filtrate was concentrated under reduced pressure. The residue was separated by chromatography on a column of silica gel with ethyl acetate\u2013hexane (1/9) as eluent. The solid product obtained was recrystallized from ethanol to afford colourless crystals (yield: 82%).3-Bromo\u00adpropane\u00adnitrile (2.0\u2005mmol) was added to a mixture of (Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019005966/lh5901sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019005966/lh5901Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019005966/lh5901Isup3.cdxSupporting information file. DOI: 1913051CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E,1\u2032E)-bis\u00ad(methanylyl\u00adidene)}bis\u00ad[2-(tri\u00adfluoro\u00admeth\u00adoxy)phenol].In the title complex, the nickel(II) ion has a square-planar coordination sphere, being ligated by two N and two O atoms of the tetra\u00addentate Schiff base ligand 6,6\u2032-{-bis(methanylyl\u00adidene)}bis\u00ad[2-(tri\u00adfluoro\u00admeth\u00adoxy)phenol]-\u03ba4O,N,N\u2032,O\u2032)nickel(II), [Ni(C18H12F6N2O4)], the nickel(II) ion has a square-planar coordination sphere, being ligated by two N and two O atoms of the Schiff base ligand 6,6\u2032-{-bis\u00ad(methanylyl\u00adidene)}bis\u00ad[2-(tri\u00adfluoro\u00admeth\u00adoxy)phenol] (L). Inversion-related mol\u00adecules are linked by a short Ni\u22efNi inter\u00adaction of 3.2945\u2005(6)\u2005\u00c5 forming a dimer. In the crystal, the dimers stack up the a axis, with a closest Ni\u22efNi separation of ca 3.791\u2005\u00c5. There are no other significant inter\u00admolecular inter\u00adactions present. However, the Hirshfeld surface analysis and the two-dimensional fingerprint plots indicate that the packing is dominated by H\u22efF/F\u22efH, H\u22efH, O\u22efH/H\u22efO and C\u22efH/H\u22efC contacts.In the title complex, (6,6\u2032-{(1 Their \u2014OH and C=N groups are involved in the formation of covalent bonding with the metal atom; besides, these mol\u00adecules are known to be easy to synthesize giving a high yield under mild conditions by solvent or solvent-free methods \u2005\u00c5 . The nickel ion Ni1 is coordinated by two imine N atoms, N6 and N7, and by two phenoxo O atoms, O2 and O3, of the tetra\u00addentate Schiff base ligand L. The bond lengths, Ni\u2014O2 and Ni\u2014O3 , and Ni\u2014N6 and Ni\u2014N7 are close to the values observed for nickel complexes of similar ligands (see section Database survey). The coordinating atoms, N6, N7, O2, O3, are essentially planar with no atom deviating from its mean plane by more than 0.0325\u2005\u00c5. The \u03c44 factor for four-coordinated metal atoms is = 0.04, indicating an almost perfect square-planar coordination sphere for atom Ni1 a-axis direction with a Ni1i\u22efNi1ii separation of ca. 3.791\u2005\u00c5 . There are no other significant inter\u00admolecular inter\u00adactions present; both C\u2014H\u22efF and C\u2014H\u22efO inter\u00adactions exceed the sum of their van der Walls radii.In the crystal, the dimers stack up the et al., 2016et al. (1970supporting information files S1(H), S2(OMe) and S3(OEt)]. A common feature of these complexes is the dimer formation with an Ni\u22efNi separation of between ca 3.2 to 3.9\u2005\u00c5. The same dimeric arrangement is found in the title complex, where this separation is 3.2945\u2005(6)\u2005\u00c5. In the majority of these complexes, the Ni\u2014Nimine bond lengths vary from ca 1.837 to 1.956\u2005\u00c5 while the Ni\u2014Ophenoxo bond lengths vary from ca 1.834 to 1.936\u2005\u00c5. In the title complex, the Ni\u2014Nimine [1.839\u2005(3) and 1.843\u2005(3)\u2005\u00c5] and Ni\u2014Ophenoxo [1.840\u2005(2) and 1.845\u2005(2)\u2005\u00c5] bond lengths fall within these limits.A search of the Cambridge Structural Database , O\u22efH (12.4%) and C\u22efH (11.3%) contacts. The electrostatic potential energy in the range \u22120.031 to 0.256 a.u., obtained using the STO-3G basis set at the Hartree\u2013Fock level of theory, is illustrated in Fig.\u00a06The Hirshfeld surface analysis , was synthesized by condensation of 2-hy\u00addroxy-3-tri\u00adfluoro\u00admeth\u00adoxy\u00adbenzaldehyde (0.0095\u2005mmol) and 1,2-ethanedi\u00adamine (0.0095\u2005mmol) in ethanol under reflux for ca 18\u2005h. The yellow product obtained was washed with ether and dried at room temperature. Ni(CH3COO)2\u00b74H2O (0.0080\u2005mmol) dissolved in 20\u2005ml of ethanol was added slowly to an ethanol (20\u2005ml) solution of L (0.0080\u2005mmol) and the mixture was refluxed for ca 6\u2005h. The orange product obtained was filtered off and washed with toluene. Red rod-like crystals of the title complex were obtained by slow evaporation of a solution in ethanol at room temperature .The title Schiff base ligand (Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2056989019001919/su5478sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989019001919/su5478Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019001919/su5478sup3.pdfCSD search S1. DOI: 10.1107/S2056989019001919/su5478sup4.pdfCSD search S2. DOI: 10.1107/S2056989019001919/su5478sup5.pdfCSD search S3. DOI: 1890705CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A new quinoxalin-derived ene-di\u00adthio\u00adcarbonate was synthesized and structurally analysed. The synthetic procedure and the compound\u2019s spectroscopic and analytical characterization are reported. I, C13H10N2O2S2, crystallizes in the monoclinic space group C2/c with eight mol\u00adecules in the unit cell. Excluding for the ethyl substituent, the mol\u00adecule of I adopts a nearly coplanar conformation (r.m.s. deviations is 0.058\u2005\u00c5), which is supported by the intra\u00admolecular C\u2014H\u22efO hydrogen-bonding inter\u00adaction between the two ring systems [C\u22efO = 2.859\u2005(3)\u2005\u00c5]. In the crystal, the mol\u00adecules form dimeric associates via two bifurcated C\u2014H\u22efO hydrogen-bonding inter\u00adactions between an ene hydrogen atom and a carbonyl functional group of an adjacent mol\u00adecule [C\u22efO = 3.133\u2005(3)\u2005\u00c5] and vice versa. The crystal structure is further stabilized by a three-dimensional network of weak hydrogen bonds between one mol\u00adecule and six adjacent mol\u00adecules as well as offset \u03c0\u2013\u03c0 stacking. The combination of the quinoxaline 2(1H)-one moiety with the di\u00adthio\u00adcarbonate moiety extends the aromaticity of the quinoxaline scaffold towards the substituent as well as influencing the \u03c0-system of the quinoxaline. The title compound is the direct precursor for a di\u00adthiol\u00adene ligand mimicking the natural cofactor ligand molybdopterin.The title compound As a result of their unusual redox and structural characteristics and those of their metal complexes, they immediately attracted considerable scientific inter\u00adest \u2005\u00c5 for C5] and the di\u00adthiol\u00adene ring , which are connected by the C3\u2014C4 bond [length = 1.465\u2005(3)\u2005\u00c5], are essentially coplanar, with an angle of only 4.89\u2005(12)\u00b0 between the two planes \u2005\u00c5; Table\u00a01et al., 19952 bonds and O2\u22efH2\u2014C2; D\u22efA = 3.133\u2005(3)\u2005\u00c5]. Here, the exact same atoms are involved as in the intra\u00admolecular hydrogen bond mentioned above. The respective hydrogen atom H2 is therefore bound to the ene carbon atom C2 and hydrogen bonded to the carbonyl oxygen (O2) of the quinoxaline moiety of the same mol\u00adecule as well as that of the adjacent mol\u00adecule , the quinoxalin substituent of the present mol\u00adecule is replaced by a coumarine , the ene-di\u00adthio\u00adcarbonate is replaced by an amino\u00adthia\u00adzole an overall weaker resonance in A, in which the benzene ring C\u2014C distances are all very similar (i.e. strongly resonant) whereas all other distances are of more pronounced single- and double-bond character and of less aromatic character.By bidirectional inter\u00admolecular hydrogen bonding, the title compound crystallizes as dimeric associate with the same donor and acceptor roles for both monomers , C7 [C7\u2014H7\u22efO1\u2005\u00c5], C9 , C12 and acceptor inter\u00adactions involving S2 [S2\u22efH9\u2014C9(\u2212x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a01)], O2 and O1 [O1\u22efH7\u2014C7 C\u2014H\u22efO and C\u2014H\u22efS hydrogen-bonding inter\u00adactions, forming a three-dimensional network Fig.\u00a02. In the H)-one was synthesized based on a reported literature procedure quinoxalin-2quinoxalin-2(1H)-one: To a solution of S-2--2-oxo-ethyl o-isopropyl carbonodi\u00adthio\u00adate in 250\u2005ml DCM/Et2O 1:1 at ambient temperature, H2SO4 (25.50\u2005ml) was added. The reaction mixture was stirred at room temperature for 2h. After that, the reaction was quenched by addition of 250\u2005ml of ice and the mixture was stirred for 30\u2005min. The organic phase was washed with brine and water 3 \u00d7 250\u2005ml. The solvent was reduced to 10\u2005ml in vacuo and the greenish precipitate was filtered off and washed on the filter with cold acetone 3 \u00d7 50\u2005ml. The title compound was obtained as a greenish-white powder. Single crystals suitable for X-ray analysis were obtained by slow diffusion of solvents with chloro\u00adform and Et2O Yield: 1.85g (20%).1H NMR \u03b4 8.79 ppm , 7.87 ppm , 7.61 ppm , 7.4 ppm , 4.39 ppm , 1.42 ppm . 13C NMR \u03b4 152.54 ppm, 144.75 ppm, 133.25 ppm, 132.58 ppm, 131.14 ppm, 130.32 ppm, 126.98 ppm, 124.00 ppm, 113.47 ppm, 37.47 ppm, 12.20 ppm. IR (KBr pellet): (\u03bd cm\u22121) = 3495 (br), 1734 (w), 1646 (sst), 1601 (st), 1579 (st), 1535 (st), 1463 (st), 1383 (w), 1280 (st), 1248 (w), 1216 (w), 1173 (st), 1128 (w), 1087 (w), 1045 (w), 950 (w), 892 (st), 868 (w), 825 (st), 785 (w), 758 (st), 631 (w), 554 (w), 529 (w), 467 (w), 432 (w). APCI\u2013MS (m/s) = 291 (M+ + H+). Analysis calculated for C13H10N2O2S2: C, 53.78; H 3.47; N 9.65; S 22.09. Found: C, 53.41; H 3.25; N 9.86; S 22.32.Uiso(H) = 1.5Ueq(C) for the methyl group, C\u2014H = 0.99\u2005\u00c5 with Uiso(H) = 1.2Ueq(C) for the methyl\u00adene group and C\u2014H = 0.95\u2005\u00c5 with Uiso(H) = 1.2Ueq(C) for the aromatic atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018007892/kq2022sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018007892/kq2022Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018007892/kq2022Isup3.cmlSupporting information file. DOI: 1845671CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structure of the title mol\u00adecular solvate features O\u2014H\u22efO hydrogen bonds, Br\u22efO and \u03c0\u2013\u03c0 inter\u00adactions. Hirshfeld surface analysis and fingerprint plots helped to identify the major contributors to the inter\u00admolecular inter\u00adactions. 8H4Br2O4\u00b7C2H6O2, crystallizes with one-half of a 2,5-di\u00adbromo\u00adterephthalic acid (H2Br2tp) mol\u00adecule and one-half of an ethyl\u00adene glycol (EG) mol\u00adecule in the the asymmetric unit. The whole mol\u00adecules are generated by application of inversion symmetry. The H2Br2tp mol\u00adecule is not planar, with the di\u00adbromo\u00adbenzene ring system inclined by a dihedral angle of 18.62\u2005(3)\u00b0 to the carb\u00adoxy\u00adlic group. In the crystal, the H2Br2tp and EG mol\u00adecules are linked into sheets propagating parallel to (R44 (12) and R44 (28) graph-set motifs. Br\u22efO and weak \u03c0\u2013\u03c0 stacking inter\u00adactions are also observed. Hirshfeld surface analysis was used to confirm the existence of these inter\u00adactions.The title compound, C As a result of symmetry restrictions, the EG mol\u00adecule adopts an anti-conformation with an O3\u2014C5\u2014C5i\u2014O3i torsion angle of 180\u00b0 .The structures of the mol\u00adecular components in the title compound are shown in Fig.\u00a012Br2tp and EG mol\u00adecules are linked by strong-to-medium O\u2014H\u22efO hydrogen bonds between carb\u00adoxy\u00adlic acid and alcohol OH functions to 0.9385 a.u. (blue) and the two-dimensional fingerprint plots are illustrated in Fig.\u00a04dnorm surface can be assigned to Br\u22efO contacts. Analysis of the two-dimensional fingerprint plots reveals that the H\u22efO/O\u22efH (28.8%) contacts are the dominant contributors to the Hirshfeld surface. The contribution of the Br\u22efH/H\u22efBr contacts is 22.1%, whereas Br\u22efBr contacts are negligible (0.9%). Other contacts viz. H\u22efH (17.7%), H\u22efC/C\u22efH (7.7%), Br\u22efC/C\u22efBr (7.2%), Br\u22efO/O\u22efBr (5.8%), C\u22efO/O\u22efC (4.5%), C\u22efC (3.3%) and O\u22efO (2.2%) also make significant contributions to the Hirshfeld surface.Hirshfeld surfaces ethyl\u00adidene]pyridine-4-carbohydrazonato-\u03ba2N\u2032,O}nickel(II)\u20132,5-di\u00adbromo\u00adterephthalic acid in 5\u2005ml of EG was heated (333\u2005K) to reflux for 15\u2005min. The reaction solution was held for 2\u20133\u2005h and colourless block-shaped crystals suitable for single-crystal X-ray diffraction analysis were obtained.HUiso(H) = 1.2Ueq(C). The H atoms bound to O atoms were located from difference-Fourier maps but were refined with distance restraints of O\u2014H = 0.82 \u00b1 0.02\u2005\u00c5 and Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019010260/wm5512sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019010260/wm5512Isup3.cdxSupporting information file. DOI: 10.1107/S2056989019010260/wm5512Isup4.hklStructure factors: contains datablock(s) I. DOI: 1941436CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "ANCA)\u2010associated vasculitis (AAV), S1P participated in MPO\u2010ANCA\u2010positive IgG\u2010induced glomerular endothelial cell (GEnC) activation via a S1P receptor (S1PR)\u2010dependent way. However, the downstream signalling pathways are not fully clear yet. In this study, we demonstrated that Rho guanosine triphosphatases (GTPases) signalling pathways, RhoA and Rac1 in particular, were implicated in MPO\u2010ANCA\u2010positive IgG\u2010mediated GEnCs activation enhanced by pathophysiological concentration of S1P in AAV. These results provide mechanistic insights into vascular barrier dysfunction in AAV, which may facilitate the development of effective therapies.Sphingosine\u20101\u2010phosphate (S1P) is a crucial regulator in vascular inflammation. Our recent study found that under pathophysiological concentration in active anti\u2010neutrophil cytoplasmic antibody ( S1P and its five G\u2010protein\u2010coupled receptors (GPCRs), S1PR1\u20105, are implicated in diverse vascular inflammatory conditions.Rho guanosine triphosphatases (GTPases) mediate diverse biological responses including morphogenesis, chemotaxis and cell cycle progression.22.1See the \u201cSupporting Information Data 2.2Primary human glomerular endothelial cells were cultured according to the manufacturer's instructions.2.3MPO\u2010ANCA\u2010positive IgGs and normal IgGs were prepared as previously described2.4Rac1 and RhoA activation assays were performed following the manufacturer's instructions .2.5As biomarkers of endothelial cell activation, levels of soluble vascular cell adhesion molecule\u20101(sVCAM\u20101) and intercellular adhesion molecule\u20101 (sICAM\u20101) in the GEnC supernatants were tested with commercially available ELISA kits .2.6P\u00a0<\u00a0.05 . Differences were considered statistically significant if 33.1P\u00a0<\u00a0.001 by ANOVA; 140.1\u00a0\u00b1\u00a08.9% vs 100%, P\u00a0<\u00a0.001 by ANOVA) Figure\u00a0,S3.3.2P\u00a0<\u00a0.001 by ANOVA; 113.9\u00a0\u00b1 9.4% vs 147.9\u00a0\u00b1\u00a05.2%, P\u00a0<\u00a0.001 by ANOVA; 124.7\u00a0\u00b1\u00a04.4% vs 147.9\u00a0\u00b1\u00a05.2%, P\u00a0<\u00a0.001 by ANOVA, respectively), while the S1PR5 agonist A97 up\u2010regulated the RhoA activity . By contrast, the ICAM\u20101 and VCAM\u20101 levels in the supernatants of GEnCs stimulated by S1P combined with MPO\u2010ANCA\u2010positive IgG increased significantly upon pre\u2010incubation with Rac1 antagonist NSC (Figure\u00a0Pre\u2010incubation of GEnCs with the RhoA antagonist CCG significantly decreased ICAM\u20101 and VCAM\u20101 levels in the supernatants of GEnCs stimulated by S1P plus MPO\u2010ANCA\u2010positive IgG (1352.33\u00a0\u00b1 \u00a0122.73\u00a0pg/mL vs 812.91\u00a0\u00b1\u00a025.12\u00a0pg/mL, 4In our present study, we demonstrated that under pathophysiological concentration in active AAV patients, S1P could activate both RhoA and Rac1 signalling pathways in MPO\u2010ANCA\u2010positive IgG\u2010treated GEnCs. According to Singleton et\u00a0al, RhoA and Rac1 play opposing roles in regulating endothelial barrier function in response to differential activation of S1PRs.In conclusion, our current study demonstrated that Rho GTPases signalling pathways were involved in S1P\u2010enhanced GEnCs activation with MPO\u2010ANCA\u2010positive IgG. RhoA activated by S1PR2\u20105 dominated the S1P\u2010induced GEnC activation in the presence of MPO\u2010ANCA\u2010positive IgG, while Rac1 activated by S1PR1 exerted opposite effect during this process. These findings provide us with more clues to determine the role of and Rho GTPases and S1P in the development of AAV.No conflict of interest to declare.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file.\u00a0Click here for additional data file."} +{"text": "The dihedral angles between the pyridine ring and its attached phenyl groups are 42.24\u2005(8) and 6.37\u2005(14)\u00b0. In the crystal, a system of classical O\u2014H\u22efO and O\u2014H\u22ef hydrogen bonds links the mol\u00adecules to form tube-like assemblies propagating parallel to the 23H20N2O4S, the sulfur atom is attached equatorially to the sugar ring with unequal S\u2014C bonds, viz.: S\u2014Cs = 1.808\u2005(2) and S\u2014Cp = 1.770\u2005(2)\u2005\u00c5 . The dihedral angles between the pyridine ring and its attached phenyl groups are 42.24\u2005(8) and 6.37\u2005(14)\u00b0. In the crystal, a system of classical O\u2014H\u22efO and O\u2014H\u22ef hydrogen bonds links the mol\u00adecules to form tube-like assemblies propagating parallel to the c-axis direction. Weak C\u2014H\u22efN inter\u00adactions are also observed.In the racemic title compound, C Thus, (1) reacted with (2) in KOH in acetone to give a product for which two isomeric N- or S-arabinoside structures were conceivable, corresponding to two possible modes of glycosyl\u00adation. The final deprotected product (see Scheme4) or its regioisomer pyridine-2-thione-N-arab\u00adinoside (5). Spectroscopic data cannot differentiate between these two structures.Here we report a one-step synthesis of the pyridine-2-thio\u00adarabinoside (4) as the only product in the solid state. We suggest that the 2,3,4-tri-O-acetyl-\u03b1-d-arabinopranosyl bromide (2) inter\u00adacts via a simple SN2 reaction to give the \u03b2-glycoside product (3), which after deprotection leads to the free 2-(\u03b2-d/l-arabino\u00adpyran\u00adosyl\u00adthio)-pyridine-3-carbo\u00adnitrile (4). This separates as a racemic mixture, presumably because of thermodynamic racemization during synthesis or crystallization is shown in Fig.\u00a01et al., 2017s 1.808\u2005(2) and S\u2014Cp 1.770\u2005(2)\u2005\u00c5 . The phenyl ring at C31 is approximately coplanar with the pyridyl ring, but the ring at C21 is significantly rotated (inter\u00adplanar angles = 6.4\u2005(2) and 42.24\u2005(8)\u00b0, respectively). The relative orientation of the pyridyl ring and the sugar moiety is defined by the torsion angles N1\u2014C2\u2014S1\u2014C11 9.7\u2005(2) and C2\u2014S1\u2014C11\u2014C12 162.73\u2005(12)\u00b0. The intra\u00admolecular contact O1\u2014H01\u22efS1, with H\u22efS 2.79\u2005(4)\u2005\u00c5 and an angle of 109\u2005(3)\u00b0, is probably too long and has too narrow an angle to be considered a hydrogen bond.The mol\u00adecular structure of contact of 3.2374\u2005(16)\u2005\u00c5 may play a supporting role, but is not shown explicitly.In the crystal, the mol\u00adecules are connected by two-centre O2\u2014H02\u22efO3d Table\u00a01, via thes Figs. 2 and 3 \u25b8.et al., 1993There is one other structure involving arabinose with a sulfur substituent at the C2 position; the arabinose is tri\u00adacetyl\u00adated and the sulfur atom, which is axially bonded to the sugar ring, acts as a bridge to a pyran\u00adopyrimidine ring system (Tomas H)-thione (1) in aqueous potassium hydroxide was added a solution of 2,3,4-tri-O-acetyl-\u03b1-d-arabino\u00adpyranosyl bromide (2) in acetone (30\u2005ml). The reaction mixture was stirred at room temperature until the reaction was judged complete by TLC (30\u2005min to 2\u2005h). The mixture was evaporated under reduced pressure at 313\u2005K and the residue was washed with distilled water to remove the potassium bromide. The solid was collected by filtration and crystallized from ethanol to give compound (3) in 70% yield (m. p. 440\u2013442\u2005K). Dry gaseous ammonia was then passed through a solution of the protected thio\u00adglycoside (3) (0.5\u2005g) in dry methanol (20\u2005ml) at 273\u2005K for 15\u2005min, and the mixture was stirred at 273\u2005K until the reaction was complete . The mixture was evaporated at 313\u2005K to give a solid residue, which was recrystallized from methanol solution to give compound (4) in 60% yield (m.p. 479\u2013480\u2005K), IR (KBr): 3370\u20133480 (OH); 2222 (CN) cm\u22121. 1H NMR : \u03b4 3.10\u20133.70 ; 4.81\u20135.20 ; 5.52 , 7.05\u20137.78 , 7.99 . Analysis calculated for C23H20N2O4S (420.47): C, 65.60%; H, 4.76%; N, 6.66%. Found: C, 65.48%; H, 4.84%; N, 6.41%.To a solution of the pyridine-2- with Uiso(H) = 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018007284/hb7743sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018007284/hb7743Isup2.hklStructure factors: contains datablock(s) I. DOI: 1843269CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Protein\u2010polymer conjugates show improved pharmacokinetics but reduced bioactivity and tumor penetration as compared to native proteins, resulting in limited antitumor efficacy. To address this dilemma, genetic engineering of a body temperature\u2010responsive and matrix metalloproteinase (MMP)\u2010cleavable conjugate of interferon alpha (IFN\u03b1) and elastin\u2010like polypeptide (ELP) is reported with spatiotemporally programmed two\u2010step release kinetics for tumor therapy. Notably, the conjugate could phase separate to form a depot postsubcutaneous injection, leading to 1\u2010month zero\u2010order release kinetics. Furthermore, it could selectively be cleaved by MMPs that are overexpressed in tumors to release IFN\u03b1 from ELP and thus to recover the bioactivity of IFN\u03b1. Consequently, it exhibits dramatically enhanced tumor accumulation, tumor penetration, and antitumor efficacy as compared to free IFN\u03b1 in two mouse models of melanoma and ovarian tumor. These findings may provide an intelligent technology of thermoresponsive and protease\u2010cleavable protein\u2010polymer conjugates with spatiotemporally programmed two\u2010step release kinetics for tumor treatment. In this proof\u2010of\u2010concept study, a MMP substrate (MMPS) as a linker was genetically fused to the C\u2010terminal of IFN\u03b1 and the N\u2010terminal of a body temperature\u2010responsive elastin\u2010like polypeptide (ELP(V)) to form a body temperature\u2010responsive and MMP\u2010cleavable IFN\u03b1\u2010MMPS\u2010ELP(V) conjugate Schemea. IFN\u03b1 is Scheme b. Furthes Scheme c. Indeed22.1Tt values were genetically engineered (see Supporting Information). Simultaneously, two MMP\u2010noncleavable IFN\u03b1\u2010ELP conjugates of IFN\u03b1\u2010ELP(V) and IFN\u03b1\u2010ELP(A) with distinct Tt values were prepared as controls. Specifically, the sequence of MMPS was Pro\u2010Leu\u2010Gly\u2010Leu\u2010Ala\u2010Gly (PLGLAG). The A and V in ELP(A) and ELP(V) were the X in VPGXG repeat unit. The repeat unit number per ELP chain was 90. The conjugates were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS\u2010PAGE) and further confirmed by matrix\u2010assisted laser desorption/ionization time\u2010of\u2010flight mass spectrometry (MALDI\u2010TOF\u2010MS) to have the expected molecular weights of the conjugates were determined by dynamic light scattering (DLS) to be around 11 nm, which were 3.8\u2010fold larger than that of IFN\u03b1 (2.9 nm) and IFN\u03b1\u2010ELP(V) were below the body temperature of 37 \u00b0C when the concentrations were above 1 \u00d7 10\u22126m. These data suggested that IFN\u03b1\u2010MMPS\u2010ELP(V) and IFN\u03b1\u2010ELP(V) would in situ form depots upon subcutaneous injections at high concentrations and then would slowly liberate from the depots into circulatory system at low concentrations. By contrast, the Tt values of IFN\u03b1\u2010MMPS\u2010ELP(A) and IFN\u03b1\u2010ELP(A) were well above the body temperature even if the concentrations were as high as 1000 \u00d7 10\u22126m, suggesting that they could not in situ form depots upon subcutaneous injections. The antiproliferative activities of the conjugates , 53.8 pg mL\u22121 for IFN\u03b1\u2010MMPS\u2010ELP(A), 54.6 pg mL\u22121 for IFN\u03b1\u2010ELP(V), 56.9 pg mL\u22121 for IFN\u03b1\u2010ELP(A)) were nearly the same, which were around 37% of that of IFN\u03b1 (IC50 = 20.2 pg mL\u22121) and IFN\u03b1\u2010MMPS\u2010ELP(A) with distinct s Figurea,b. The 2.2\u22121, IC50) and IFN\u03b1\u2010MMPS\u2010ELP(A) were increased from 37% to about 91% of that of IFN\u03b1 and IFN\u03b1\u2010MMPS\u2010ELP(A), the conjugates were treated with MMP\u20102, followed by SDS\u2010PAGE analysis Figure g. As exp2.3Figuret1/2) of IFN\u03b1\u2010MMPS\u2010ELP(A) (8.9 \u00b1 1.0 h) and IFN\u03b1\u2010ELP(A) (9.6 \u00b1 2.7 h) were 6.4\u2010 and 6.9\u2010fold longer than that of free IFN\u03b1 (1.4 \u00b1 0.21 h), respectively. The area under the curves (AUCs) of IFN\u03b1\u2010MMPS\u2010ELP(A) (684.1 \u00b1 12.1 \u00b5g L\u22121\u00b7h) and IFN\u03b1\u2010ELP(A) (707.3 \u00b1 37.1 \u00b5g L\u22121 h) were 11.1\u2010 and 11.4\u2010fold larger than that of free IFN\u03b1 (61.8 \u00b1 1.7 \u00b5g L\u22121\u00b7h), respectively. Similarly, the biodistribution of IFN\u03b1\u2010MMPS\u2010ELP(A) was almost the same as that of IFN\u03b1\u2010ELP(A) but much superior to that of free IFN\u03b1 , respectively. These data indicated that the introduction of the MMPS linker into IFN\u03b1\u2010ELP(A) did not change the pharmacokinetics and biodistribution significantly. However, IFN\u03b1\u2010MMPS\u2010ELP(A) showed significantly enhanced tumor penetration as compared to both IFN\u03b1\u2010ELP(A) and free IFN\u03b1, as indicated by the more intense fluorescence (yellow) observed in the area that was distant from the vessels (red) for IFN\u03b1\u2010MMPS\u2010ELP(A) than for IFN\u03b1\u2010ELP(A) and free IFN\u03b1 on its in vivo properties in a mouse model of C8161 melanoma Figure. The pha2.4\u22121 body weight) for IFN\u03b1\u2010MMPS\u2010ELP(V) was the same as that for IFN\u03b1\u2010ELP(V), which was 5.0\u2010 and 6.7\u2010fold higher than those for IFN\u03b1\u2010MMPS\u2010ELP(A) (20 mg kg\u22121 body weight) and free IFN\u03b1 (15 mg kg\u22121 body weight), respectively . Interestingly, a bump in situ formed and gradually diminished over 40 d after subcutaneous injection of IFN\u03b1\u2010MMPS\u2010ELP(V) or IFN\u03b1\u2010ELP(V) at its MTD . This phenomenon suggested that both IFN\u03b1\u2010MMPS\u2010ELP(V) and IFN\u03b1\u2010ELP(V) could in situ form depots after subcutaneous injections at their MTDs because the Tt values of the two conjugates were far below the in vivo temperature and IFN\u03b1 rapidly increased to 5878.7 \u00b1 187.1 \u00b5g L\u22121 at 4.0 h and 5456.9 \u00b1 758.5 \u00b5g L\u22121 at 2.8 h, respectively, and then decreased quickly. Representative pharmacokinetic parameters were generated by fitting the data with a one\u2010compartment model . Notably, The t1/2 of IFN\u03b1\u2010MMPS\u2010ELP(V) (422.2 \u00b1 13.7 h) was close to that of IFN\u03b1\u2010ELP(V) (491.8 \u00b1 38.1 h), but was 46.9\u2010 and 222.2\u2010fold longer than those for IFN\u03b1\u2010MMPS\u2010ELP(A) (9.0 \u00b1 0.87 h) and free IFN\u03b1 (1.9 \u00b1 0.08 h), respectively. The AUC of IFN\u03b1\u2010MMPS\u2010ELP(V) (2755.9 \u00b1 16.8 mg L\u22121\u00b7h) was close to that of IFN\u03b1\u2010ELP(V) (3102.0 \u00b1 269.8 mg L\u22121\u00b7h), but was 23.3\u2010 and 58.8\u2010fold larger than those for IFN\u03b1\u2010MMPS\u2010ELP(A) (118.3 \u00b1 15.1 mg L\u22121 h) and free IFN\u03b1 (46.9 \u00b1 7.8 mg L\u22121\u00b7h), respectively. Furthermore, the AUCs of IFN\u03b1\u2010MMPS\u2010ELP(V) and IFN\u03b1\u2010ELP(V) were correlated to time linearly , we further studied the in vivo properties of IFN\u03b1\u2010MMPS\u2010ELP(V) postsubcutaneous administration at its maximum tolerated dose (MTD). The MTD (100 mg kg\u22121 tissue) was nearly equal to that of IFN\u03b1\u2010ELP(V) (555.7 \u00b1 164.7 ng g\u22121 tissue), but was 18.0\u2010 and 317.2\u2010fold higher than those for IFN\u03b1\u2010MMPS\u2010ELP(A) (29.9 \u00b1 1.8 ng g\u22121 tissue) and free IFN\u03b1 (1.7 \u00b1 0.05 ng g\u22121 tissue) at 3 d postinjection, respectively was studied after subcutaneous administration at the MTD. The biodistribution of IFN\u03b1\u2010MMPS\u2010ELP(V) was similar to that of IFN\u03b1\u2010ELP(V), but was much better than those of IFN\u03b1\u2010MMPS\u2010ELP(A) and free IFN\u03b1 . Notably, the tumor concentration of IFN\u03b1\u2010MMPS\u2010ELP(V) was 3.5\u2010, 9.1\u2010, and 19.5\u2010fold smaller than those for IFN\u03b1\u2010ELP(V) (145.3 \u00b1 49.1 mm3), IFN\u03b1\u2010MMPS\u2010ELP(A) (376.8 \u00b1 121.7 mm3), and free IFN\u03b1 (808.2 \u00b1 226.1 mm3) at 30 d after the subcutaneous injections, respectively. Notably, 60% of the mice in the IFN\u03b1\u2010MMPS\u2010ELP(V) treatment group were tumor free, which was much higher than those for the treatments with IFN\u03b1\u2010ELP(V) (30%), IFN\u03b1\u2010MMPS\u2010ELP(A) (0%), and free IFN\u03b1 (0%) was 3.4\u2010, 6.0\u2010, and 11.6\u2010fold smaller than those for IFN\u03b1\u2010ELP(V) (250.4 \u00b1 24.9 mm3), IFN\u03b1\u2010MMPS\u2010ELP(A) (434.9 \u00b1 119.9 mm3), and free IFN\u03b1 (847.9 \u00b1 248.2 mm3) at 33 d after the subcutaneous injections, respectively. Notably, 37.5% of the mice in the IFN\u03b1\u2010MMPS\u2010ELP(V) treatment group were tumor free, whereas the tumor cure percentages for the treatments with IFN\u03b1\u2010ELP(V), IFN\u03b1\u2010MMPS\u2010ELP(A), and free IFN\u03b1 were 0% in a mouse model of melanoma. After subcutaneous injection at its MTD, IFN\u03b1\u2010MMPS\u2010ELP(V) suppressed melanoma growth more efficiently than IFN\u03b1\u2010ELP(V), IFN\u03b1\u2010MMPS\u2010ELP(A), and free IFN\u03b1 Figurea. The av2.6FigureAll the treatments did not cause obvious loss of body weight . As indicated by hematoxylin and eosin (H&E) staining, a more extensive degree of kidney damage was observed in the IFN\u03b1 treatment than in the other treatments, but no histological change in other major organs was observed in all the treatments Figure. This re3thuman = tmouse (Whuman/Wmouse)0.13 in which thuman and tmouse are the half\u2010lives of the conjugate in human and mouse, respectively, and Whuman and Whuman are the body weights of human and mouse, respectively.In conclusion, we have reported genetic engineering of a body\u2010temperature\u2010responsive and MMP\u2010cleavable protein\u2010polymer conjugate of IFN\u03b1\u2010MMPS\u2010ELP(V) with not only dramatically improved pharmacokinetics but also remarkably enhanced tumor accumulation and penetration, and antitumor efficacy. IFN\u03b1\u2010MMPS\u2010ELP(V) can be genetically engineered to own dual functions of body temperature responsiveness and MMP cleavability that enable spatiotemporally programmed two\u2010step release kinetics. The body temperature responsiveness can remarkably enhance the tolerability, pharmacokinetics, and tumor accumulation of the conjugate through the mechanism of body temperature\u2010responsive controlled release due to the concentration dependence of the phase transition temperature. Notably, in a mouse model, subcutaneous injection of the conjugate at its MTD offers an extremely prolonged circulation half\u2010life of 422.2 h. The half\u2010life of the conjugate in human is expected to be as long as 1195.5 h according to the empirical formulus of The authors declare no conflict of interest.SupplementaryClick here for additional data file."} +{"text": "The structure of a deca\u00adahydro\u00adacridine derivative with phenyl substituents at the 3- and 5-positions of the cyclo\u00adhexenone rings is reported. An extensive range of O\u2014H\u22efO, C\u2014H\u22efO hydrogen bonds augmented by C\u2014H\u22ef\u03c0(ring) hydrogen bonds an O\u22efBr halogen bond and an unusual Br\u22ef\u03c0(ring) contact stabilizes the crystal packing. 33H29BrClNO4, (I), the hexa\u00adhydro-2H-acridine ring system has a hy\u00addroxy\u00adethyl substituent on the N atom and a 3-bromo-6-chloro-2-hy\u00addroxy\u00adphenyl substituent on the central C atom at the 9-position. An unusual feature of the mol\u00adecule is that the substituents at the 3- and 5-positions of the outer cyclo\u00adhexenone rings are phenyl rings rather than the more common dimethyl substituents. C atoms on both of the cyclo\u00adhexenone rings are disordered over two sites. In the crystal structure, O\u2014H\u22efO, C\u2014H\u22efO and C\u2014H\u22ef\u03c0(ring) hydrogen bonds combine with an Br\u2014O and unusual C\u2014Br\u22ef\u03c0(ring) halogen bonds to generate a three dimensional network with mol\u00adecules stacked along the a-axis direction.In the structure of the title compound C These include compounds that are used as anti-infammatory ring. The C2 and C3 atoms of one cyclo\u00adhexenone are disordered over two sites as is the C6 atom of the corresponding cyclo\u00adhexenone. Their occupancy ratios refine to 0.521\u2005(10):0.479\u2005(10) for C2,C3 and 0.746\u2005(9):0.254\u2005(9) for C6. Only details of the major disorder components will be considered here. The central C9,N10,C11\u2013C14 ring adopts a half-chair conformation and is inclined to the adjacent C1\u2013C4,C11,C12 and C5\u2013C8,C13,C14 rings at angles of 7.11\u2005(18) and 21.64\u2005(10)\u00b0, respectively, so the hexa\u00adhydro-2H-acridine unit is far from planar. The 3-bromo-6-chloro-2-hy\u00addroxy\u00adphenyl ring subtends an angle of 84.39\u2005(6)\u00b0 to this central ring. The C1\u2013C4,C11,C12 ring is best described as a severely flattened boat while the C5\u2013C8,C13,C14 system is in a distorted half-chair conformation. The phenyl substituents on these outer cyclo\u00adhexenone rings are inclined to their parent rings at angles of 76.87\u2005(12)\u00b0 for C31\u2013C36 and 86.27\u2005(8)\u00b0 for C61\u2013C66. The N-bound 2-hy\u00addroxy\u00adethyl substituent points away from the convex face of the hexa\u00adhydro-2H-acridine system as does the 3-bromo-6-chloro-2-hy\u00addroxy\u00adphenyl substituent.The title compound (I)ly Fig.\u00a01. The cenO\u22efO8 hydrogen bonds, Table\u00a01C(9) chains along the b-axis direction, linking the mol\u00adecules in a head-to-tail fashion, Fig.\u00a02a-axis direction through C65\u2014H65\u22efCg7 contacts, Fig.\u00a03A\u22efO16 hydrogen bonds form inversion dimers that enclose Cg8 inter\u00adactions. Adjacent dimers are linked by C34\u2014H34\u22efCl5\u2032 hydrogen bonds, forming double chains of mol\u00adecules along the ab diagonal, Fig.\u00a04v halogen bonds \u2005\u00c5, C3\u2032\u2014Br3\u2032\u22efCg4 = 83.89\u2005(7)\u00b0; Cg4 is the centroid of the C1\u2032\u2013C6\u2032 benzene ring] et al. 2016H-acridine ring system, emphasizing the uniqueness of the structure reported here. Refining the search to structures with CH2CH substitution on the acridine N atom reduced the hits to seven, four of which have hy\u00addroxy\u00adethyl substituents on N10 = 0.95\u2005\u00c5 for aromatic, 0.99\u2005\u00c5 for methyl\u00adene and 1.00\u2005\u00c5 for methine H atoms, all with Uiso = 1.2Ueq(C). The C2 and C3 atoms in the C1\u2013C4,C11,C12 cyclo\u00adhexenone ring and atom, C6, in the corresponding C5\u2013C8,C13,C14 ring are disordered over two positions. Their occupancies were refined to sum to unity with the disordered atoms of the different rings allowed to refine separately. The occupancies converged to ratios of 0.521\u2005(10): 0.479\u2005(10) for C2 and C3 and 0.746\u2005(9): 0.254\u2005(9) for C6. Positions of the hydrogen atoms on adjacent methyl\u00adene groups and phenyl rings were assigned taking this disorder into account but a somewhat close H15A\u22efH5C contact was still observed. One reflection with Fo >>> Fc, was omitted from the final refinement cycles.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018010873/ff2154sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018010873/ff2154Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018010873/ff2154Isup3.cmlSupporting information file. DOI: 1859007CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "C\u2014H\u22efO, C\u2014F\u22ef\u03c0 and F\u22efF inter\u00adactions are also observedThe title compound crystallizes with four mol\u00adecules in the unit cell (Z = 4) and one formula unit in the asymmetric unit. In the crystal, mol\u00adecules are linked in a head-to-tail fashion into dimers along the b-axis direction through weak C\u2014H\u22efBr and C\u2014O\u22efC 10H5BrF6O, synthesized via continuous stirring of 3,5-bis\u00ad(tri\u00adfluoro\u00admeth\u00adyl) aceto\u00adphenone with bromine in an acidic medium and concentrated under reduced pressure, crystallizes with four mol\u00adecules in the unit cell (Z = 4) and one formula unit in the asymmetric unit. In the crystal, mol\u00adecules are linked in a head-to-tail fashion into dimers along the b-axis direction through weak C\u2014H\u22efBr and C\u2014O\u22efCsp2 inter\u00adactions. C\u2014H\u22efO, C\u2014F\u22ef\u03c0 and F\u22efF inter\u00adactions are also observed.The title compound, C N-bromo\u00adsuccinimide . The torsion angle between the alkyl bromide unit and the phenyl ring (C3\u2014C2\u2014C1\u2014Br1) is \u2212179.6\u2005(3)\u00b0 whereas that between the alkyl bromide and carbonyl parts (O1\u2014C2\u2014C1\u2014Br1) is 0.3\u2005(5)\u00b0, which shows a preference for a syn orientation of the alkyl bromide unit with respect to the carbonyl group.A displacement ellipsoid plot of the title compound with the atom labelling is shown in Fig.\u00a01c-axis direction by C\u2014H\u22efO and C\u2014F\u22ef\u03c0 inter\u00adactions [2.868\u2005(4)\u2005\u00c5] inter\u00adactions, resulting in a bilayer which further packs in parallel fashion along the a-axis direction (O\u22ef\u03c0 = 3.252\u2005\u00c5) inter\u00adactions. The dimers are linked along the s Table\u00a01. The asson Fig.\u00a03.et al., 2016et al., 2011et al., 1991Z = 4, features two prominent hydrogen bonds involving the oxygen atom while in the second, also Z = 4, the oxygen atom forms a complex with anti\u00admony penta\u00adchloride.There are more than 1000 crystal structure of phenyl ethanone derivatives in the Cambridge Structural Database (CSD) : 8.44 , 8.13 , 4.48 ; 13C NMR: : 188.81, 135.31, 133.06, 132.83, 132.60, 128.99, 127.08, 127.06, 125.42, 123.61, 121.80, 120.00, 29.46.A stirred solution of 3,5-bis\u00ad(tri\u00adfluoro\u00admeth\u00adyl) aceto\u00adphenone in acetic acid (5\u2005mL) was added dropwise to bromine in acetic acid. The reaction medium was stirred at room temperature for 5\u2005h. To the resulting mixture, water (5\u2005mL) was added and the mixture was concentrated under reduced pressure. The residue obtained was diluted with ethyl\u00adacetate (10\u2005mL), the organic layer washed with water (10\u2005mL) and a sodium bicarbonate solution (5\u2005mL), and filtered through dried sodium sulfate and evaporated to obtain 1-phen\u00adyl)-2-bromo\u00adethanone as a light-yellow solid in 62% yield. m.p: 317\u2013318\u2005K. Uiso(H) = 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018007478/ds2250sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018007478/ds2250Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018007478/ds2250Isup3.cmlSupporting information file. DOI: 1843826CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "One chain type, an unanchored NEDD8 trimer, specifically bound to the second zinc finger domain of PARP\u20101 and attenuated its activation. In cells in which Nedp1 is deleted, large amounts of tri\u2010NEDD8 constitutively form, resulting in inhibition of PARP\u20101 and protection from PARP\u20101\u2010dependent cell death. Surprisingly, these NEDD8 trimers are additionally acetylated, as shown by mass spectrometry analysis, and their binding to PARP\u20101 is reduced by the overexpression of histone de\u2010acetylases, which rescues PARP\u20101 activation. Our data suggest that trimeric, acetylated NEDD8 attenuates PARP\u20101 activation after oxidative stress, likely to delay the initiation of PARP\u20101\u2010dependent cell death.NEDD8 is a ubiquitin\u2010like protein that activates cullin\u2010RING E3 ubiquitin ligases (CRLs). Here, we identify a novel role for NEDD8 in regulating the activity of poly(ADP\u2010ribose) polymerase 1 (PARP\u20101) in response to oxidative stress. We show that treatment of cells with H UBLs are small proteins of approximately 8\u00a0kDa molecular mass that become covalently linked to other proteins via an isopeptide bond, usually on lysine residues polymerase 1 (PARP\u20101) and attenuates its activation. PARP\u20101 overactivation mediates cell death after oxidative stress, and our data suggest that tri\u2010NEDD8 prevents PARP\u20101 hyperactivation to delay premature commitment to cell death.et\u00a0al, et\u00a0al, Nedp1 led to the accumulation of neddylated species that do not migrate at the ~\u00a0100\u00a0kDa size of neddylated cullins in both cell lines . When exposed to cell extract, this resin efficiently enriched neddylated proteins from both NEDP1 KO cells and from WT cells, but it did not enrich for ubiquitylated proteins and the NEDD8 E2 enzyme, UBE2M, but in the absence of any NEDD8 E3. The smallest protein in these in\u00a0vitro reactions, aside from NEDD8 (8.5\u00a0kDa), was UBE2M, with a molecular mass of ~\u00a022\u00a0kDa. The smallest possible, non\u2010NEDD8\u2010NEDD8 conjugate in these reactions, a UBE2M\u2010NEDD8 linkage, would thus have a molecular mass of ~\u00a031\u00a0kDa. However, while this product was present in the reaction, conjugates were also formed at lower molecular masses that could only correspond to unanchored NEDD8 dimers, trimers and tetramers inhibitor (MLN4924) alone, and not following treatment with the ubiquitin E1 enzyme (UBE) inhibitor (MLN7243) ation (PAR), including the poly(ADP\u2010ribose) polymerase PARP\u20101 Fig\u00a0. In factet\u00a0al, et\u00a0al, +) to covalently attach ADP\u2010ribose to various amino acids, including aspartic acid, glutamic acid, lysine, arginine and serine, that are present on itself and on its substrate proteins ation are known regulators of DNA repair and of the oxidative stress response. PARP\u20101 senses DNA damage caused by oxygen radicals and aids in the early steps of DNA repair , together with its substrate receptor Kelch\u2010like ECH\u2010associated protein 1 (Keap1), is involved in the cellular response to oxidative stress. Oxidative stress inhibits the Cul3/Keap1\u2010dependent degradation of the transcription factor nuclear factor erythroid 2\u2010related factor 2 (Nrf2), which drives gene expression that protects against cell death , but not to its automodification and catalytic domain Fig\u00a0G. The DNet\u00a0al, In addition to its roles in DNA repair, PARP\u20101 also inhibits the transcription of some NF\u2010\u03baB\u2010responsive genes, including CXCL10, independently of its catalytic activity, but via its ability to directly bind to DNA Fig\u00a0C to puriFrom these results, we hypothesized that the acetylation of NEDD8 may serve to mask the positively charged lysine residues of NEDD8 to facilitate its binding to the PARP\u20101 Zn2 domain. To confirm whether changes in acetylation influence the interaction of NEDD8 with PARP\u20101, we overexpressed the de\u2010acetylases HDAC1 or HDAC2 in NEDP1 KO cells, which indeed led to a decrease in the amount of NEDD8 trimer bound to exogenously expressed and precipitated GFP\u2010PARP\u20101 Zn1\u00a0+\u00a02 Fig\u00a0C. Likewiin\u00a0vitro NEDD8 chains by NEDP1 was strongly inhibited by the presence of even low amounts of H2O2 that occur following oxidation of DNA. Zn2 binds to DNA with the highest affinity , Sh \u03b1\u2010NEDP1 1:1,000 (DSTT S378D against AA1\u2013212), Ms \u03b1\u2010DCUN1D1 1:1,000 (Sigma Clone 3D7), Sh \u03b1\u2010DCUN1D2 1:1,000 (DSTT S995C against AA1\u201360) incubated with recombinant DCNL1 (AA 1\u201345) peptide to block cross reaction, Sh \u03b1\u2010DCUN1D3 1:2,000 (DSTT S996C against AA1\u201360), Sh \u03b1\u2010DCUN1D4 1:2,000 (DSTT S997C against AA1\u2013119), Sh \u03b1\u2010DCUN1D5 1:2,000 (DSTT S998C against AA1\u2013124), Rb \u03b1\u2010NEDD8 1:1,000 (Abcam ab81264), Sh \u03b1\u2010UBE2M 1:1,000 [DSTT S432D against full length (mouse)], Sh \u03b1\u2010UBE2F 1:1,000 (DSTT S438D against AA1\u201370), Ms \u03b1\u2010Actin 1:1,000 , Rb \u03b1\u2010RBX1 1:200 (Thermo PA5\u201016282), Rb \u03b1\u2010CAND1 1:1,000 (CST 7433S), Rb \u03b1\u2010CSN5 1:1,000 (Abcam ab12323), Rb \u03b1\u2010CSN8 1:1,000 (Abcam EPR5139), Rb \u03b1\u2010UBA3 1:1,000 (Epitomics 5157\u20101), Ms \u03b1\u2010AppBp1/ULA1 1:1,000 (BD 611865), Rb \u03b1\u2010Cul1 1:1,000 (Life Tech 718700), Rb \u03b1\u2010Cul2 1:5,000 (Life Tech 700179), Sh \u03b1\u2010Cul3 1:1,000 , Sh \u03b1\u2010Cul3 1:1,000 (DSTT S464D AA544\u2013768), Sh \u03b1\u2010Cul4A 1:3,500 (DSTT S087D AA1\u2013124), Rb \u03b1\u2010Cul4A 1:1,000 (CST 2699S), Sh \u03b1\u2010Cul4B 1:1,000 (DSTT S070D AA1\u2013162), Sh \u03b1\u2010Cul5 1:1,000 (DSTT S073D AA577\u2013689), Ms \u03b1\u2010I\u03baB\u03b1 1:1,000 (CST 4814), Ms \u03b1\u2010pI\u03baB\u03b1 1:1,000 (CST 9246 clone 5A5), Ms \u03b1\u2010PAR Polymer 1:1,000 (Enzo Lifescience ALX\u2010804\u2010220\u2010R100 clone 10H), Rb \u03b1\u2010PARP\u20101 1:1,000 (CST 9532), Rb \u03b1\u2010AIF 1:1,000 (Abcam ab 32516), Rb \u03b1\u2010NRF2 1:1,000 (Abcam ab62352), Rb \u03b1\u2010HIF\u20101\u03b1 1:1,000 , Ms \u03b1\u2010FLAG M2 1:2,000 (Sigma F3165), Ms \u03b1\u2010GFP 1:5,000 (Abcam ab184519); Rb \u03b1\u2010PARG 1:1,000 (CST 66564S); Rb \u03b1\u2010Keap1 1:1,000 (CST 8047).2O2, Sigma (216763); Camptothecin, Sigma (C9911); Cycloheximide, Sigma (C1988); DPQ, Calbiochem (300270); CellTiter\u2010Glo, Promega (G7570); Olaparib, Cambridge Biosciences (CAY10621); PARG inhibitor PDD 17273, Tocris (5952); Sodium Butyrate, Sigma (303410); GFP\u2010Trap_A, Chromotek (gta), PARP\u20101\u2010Trap_A, Chromotek (xta); phosSTOP, Sigma (4906845001); cOmplete EDTA\u2010free protease inhibitor cocktail (Roche\u201011836170001); Pepstatin A, Sigma (P5318), Bestatin hydrochloride, Sigma (B8385); Deoxyribonuclease I, Sigma (D5025); LDS Sample Buffer, Life Technologies (NP0007); Colloidal Coomassie, Expedeon (ISB1L); IPTG, Formedium (IPTG025); Micrococcal Nuclease, NEB (M0247S).TNF\u2010\u03b1, PeproTech (300\u201001A); HALO\u2010link resin, Promega (G1913); MLN7243, Active Biochemicals (A\u20101384); MLN4924, Active Biochemicals (A\u20101139); Hhis6\u2010HALO NEDP1(C163A) DSTT DU28042his6\u2010HALO NEDP1(C163A D29W A98K G99K) DSTT DU28008pCMV5\u2010NEDP1 DSTT DU28199GST\u2010NEDP1 DSTT DU23262Frt/TO N\u2010FLAG empty DSTT DU13236Frt/TO N\u2010GFP empty DSTT DU45825Frt/TO C\u2010GFP empty DSTT DU41574Frt/TO C\u2010FLAG empty DSTT DU43547GST empty DSTT DU49206et\u00a0al, gRNAs for NEDP1 and CAND1 were generated by mutagenesis PCR of pEsgRNA (Munoz NEDP1 gRNA F 5\u2032\u2010GGAAAGGACGAAACACCGCCCCGTAGTCTTGAGTTACAGTTTTAGAGCTAGAAAT\u20103\u2032NEDP1 gRNA R5\u2032\u2010ATTTCTAGCTCTAAAACTGTAACTCAAGACTACGGGGCGGTGTTTCGTCCTTTCC\u20103\u2032CAND1 gRNA F 5\u2032\u2010GGAAAGGACGAAACACCTCACCTAAAGTCCTTGTCGCGTTTTAGAGCTAGAAAT\u20103\u2032CAND1 gRNA R 5\u2032\u2010ATTTCTAGCTCTAAAACGCGACAAGGACTTTAGGTGAGGTGTTTCGTCCTTTCC\u20103\u2032Full\u2010length PARP\u20101 (NM_001618.3) was amplified from HUVEC cDNA and cloned into the N\u2010terminal GFP\u2010tagged vector. For domain mapping, individual zinc finger domains were cloned into the C\u2010terminal GFP vector and the catalytic domain was cloned into the N\u2010terminal GFP vector. GST\u2010PARP\u20101 ZN1\u00a0+\u00a02 was cloned in pGEX\u20106P\u20102 (GE Lifesciences). NSUN2 (NM_017755.5) was amplified from HUVEC cDNA and cloned into the N\u2010terminal FLAG empty vector. HDAC1 (NM_004964.2) and HDAC2 (NM_001527.3) were both amplified from HUVEC cDNA and cloned into the C\u2010terminal FLAG empty vector.PARP\u20101 Full(NM_001618.3) F 5\u2032\u2010ATGGCGGAGTCTTCGGATAAGCTCT\u20103\u2032PARP\u20101 Full(NM_001618.3) R 5\u2032\u2010TAGTACGCGGCCGCTTACCACAGGGAGGTCTTAAAATTGAATTTCAGTTTCAGC\u20103\u2032PARP\u20101 Zn1 (AA1\u201096) F 5\u2032\u2010ATGGCGGAGTCTTCGGATAAGCTCT\u20103\u2032PARP\u20101 Zn1 (AA1\u201096) R 5\u2032\u2010GTACTAGCGGCCGCGCCTGTCACTCCTCCAGCTTCCG\u20103\u2032PARP\u20101 Zn2 (AA97\u2010215) F 5\u2032\u2010TAGTACGGATCCAAAGGCCAGGATGGAATTGGTAGC\u20103\u2032PARP\u20101 Zn2 (AA97\u2010215) R stop 5\u2032\u2010GTACTAGCGGCCGCCTATCCATCCACCTCATCGCCTTTTCT\u20103\u2032PARP\u20101 Zn2 (AA97\u2010215) R no stop 5\u2032\u2010GTACTAGCGGCCGCTCCATCCACCTCATCGCCTTTTCT\u20103\u2032PARP\u20101 Zn3 (AA216\u2010336) F 5\u2032\u2010TAGTACGGATCCGTGGATGAAGTGGCGAAGAAGAAATCTA\u20103\u2032PARP\u20101 Zn3 (AA216\u2010336) R 5\u2032\u2010GTACTAGCGGCCGCTGGGGTTACCCACTCCTTCC\u20103\u2032PARP\u20101 C\u2010terminal (AA336\u20101014) F 5\u2032\u2010TAGTACCCCGGGAAGGAATTCCGAGAAATCTCTTACCTCAA\u20103\u2032PARP\u20101 C\u2010terminal (AA336\u20101014) R5\u2032\u2010TAGTACGCGGCCGCTTACCACAGGGAGGTCTTAAAATTGAATTTCAGTTTCAGC\u20103\u2032NSUN2(NM_017755.5) F 5\u2032\u2010AATAATAGAATTCATGGGGCGGCGGTCGCGGG\u20103\u2032NSUN2(NM_017755.5) R 5\u2032\u2010TATTATTGCGGCCGCTCACCGGGGTGGATGGACCCCC\u20103\u2032HDAC1(NM_004964.2) F 5\u2032\u2010AAAGGATCCATGGCGCAGACGCAGGGCACCCGGAGGAAAGT\u20103\u2032HDAC1(NM_004964.2) R 5\u2032\u2010TACGCGGCCGCGGCCAACTTGACCTCCTCCT\u20103\u2032HDAC2(NM_001527.3) F 5\u2032\u2010TACGGATCCATGGCGTACAGTCAAGGAGGCGGCAAAAAA\u20103\u2032HDAC2(NM_001527.3) R 5\u2032\u2010TACGCGGCCGCGGGGTTGCTGAGCTGTTCTGATTTGGTTC\u20103\u2032Ct\u2212\u2206\u2206 method for 4\u00a0h at 37\u00b0C and harvested with RNeasy following the manufacturer's protocol. Following RNA purification, RNA concentration was measured with a NanoDrop (Thermo Fisher Scientific) and 1\u00a0\u03bcg of RNA was used to generate cDNA using the High\u2010Capacity cDNA Reverse Transcription Kit (Invitrogen #4368814). qPCRs were performed with Brilliant III Ultra\u2010Fast SYBR Green qPCR master mix (Agilent 600882) on a Bio\u2010Rad CFX96. The 218S F 5\u2032\u2010GTAACCCGTTGAACCCCATT\u20103\u203218S R 5\u2032\u2010CCATCCAATCGGTAGTAGCG\u20103\u2032CXCL10 F 5\u2032\u2010GCTGATGCAGGTACAGCGT\u20103\u2032CXCL10 R 5\u2032\u2010CACCATGAATCAAACTGCGA\u20103\u2032I\u03baB\u03b1 F 5\u2032\u2010GATCCGCCAGGTGAAGGG\u20103\u2032I\u03baB\u03b1 R 5\u2032\u2010GCAATTTCTGGCTGGTTGG\u20103\u2032Nrf2 F 5\u2032\u2010AGACGGTATGCAACAGGACA\u20103\u2032Nrf2 R 5\u2032\u2010AGTTTGGCTTCTGGACTTGGA\u20103\u2032NQO1 F 5\u2032\u2010TCACCGAGAGCCTAGTTCCG\u20103\u2032NQO1 R 5\u2032\u2010TGGCATAGTTGAAGGACGTCC\u20103\u2032GAPDH F 5\u2032\u2010GAAATCCCATCACCATCTTCCAGG\u20103\u2032GAPDH R 5\u2032\u2010GTACCTCTTCCGACCCCGAG\u20103\u2032et\u00a0al . GST\u2010NEDP1 and GST vectors were transformed into BL21 cells and purified with Glutathione Sepharose 4B . His6\u2010USP1/UAF1 was obtained from the Division of Signal Transduction Therapy (DSTT) product, DU23056. The plasmids encoding GST\u2010PARP\u20101 Zn1\u00a0+\u00a02, GST\u2010PARP\u20101 Zn1\u2010GFP and GST\u2010PARP\u20101 Zn2\u2010GFP were transformed into BL21 Rosetta 2 (DE3)\u2010competent cells (Novagen 71400). Cells were grown in 2YT media supplemented with 100\u00a0\u03bcM ZnSO4, and protein expression was induced with 0.2\u00a0mM IPTG at 16\u00b0C for 20\u00a0h. Cells were pelleted by centrifugation, and pellets were resuspended in lysis buffer . Pellets were sonicated; then, 5\u00a0mM MgCl2 and DNase I were added to 200\u00a0Kunitz/ml; and the cell suspension was incubated with rotation at 4\u00b0C for 1\u00a0h. Cell lysate was spun at 20,000\u00a0\u00d7\u00a0g for 30\u00a0min, and supernatant was collected. GST fusion proteins were then purified with Glutathione Sepharose 4B . The GST tag was cleaved from GST\u2010PARP\u20101 Zn1\u2010GFP and GST\u2010PARP\u20101 Zn2\u2010GFP with PreScission Protease . Bound DNA was removed from Zn1\u2010GFP and Zn2\u2010GFP via a Heparin column as previously described in a 37\u00b0C incubator with 5% CO2. Cell lines were routinely checked for\u00a0mycoplasma contamination. siRNA\u2010mediated knockdown of DCNL1\u20105 was carried out as previously described supplemented with 10% FBS and 3\u00a0mM l, et\u00a0al and Munol . To immunoprecipitate the FLAG\u2010tagged proteins, cells were harvested as above in buffer A. Supernatants were incubated with anti\u2010FLAG\u2010M2 magnetic beads (Sigma) for 3\u00a0h at 4\u00b0C. Beads were then washed three times with 0.5\u00a0ml of buffer A without inhibitors and then resuspended in reaction buffer . Beads were split evenly between three tubes, and either GST\u2010NEDP1, his6\u2010USP1/UAF1 or no protein was added to each tube. Reactions were then incubated at 30\u00b0C for 30\u00a0min before being stopped with LDS sample loading buffer. To immunoprecipitate endogenous PARP\u20101 and GFP fusion proteins, cells were harvested with buffer B: 50\u00a0mM Tris\u2013HCl pH 7.5, 150\u00a0mM NaCl, 5% glycerol, 0.5% NP\u201040, 5\u00a0mM MgCl2, 0.2\u00a0mM CaCl2, 1\u00a0\u03bcM pepstatin A, 1\u00a0\u03bcM bestatin supplemented with phosStop and cOmplete mini protease inhibitor. Lysates were sonicated for six pulses of 0.5\u00a0s. Subsequently, DNase I (200\u00a0Kunitz/ml) was added to lysates and incubated for 1\u00a0h at 4\u00b0C with rotation. Lysates were centrifuged at 4\u00b0C for 20\u00a0min at 17,000\u00a0\u00d7\u00a0g, and supernatants were collected. GFP\u2010Trap beads or PARP\u20101\u2010Trap beads were incubated with lysates for 3\u00a0h at 4\u00b0C with rotation. Next, agarose beads were washed three times with 0.5\u00a0ml of wash buffer . Bound proteins were eluted with LDS sample loading buffer and heated at 95\u00b0C for 5\u00a0min with 10\u00a0mM DTT. For GFP pulldown with HDAC overexpression, cells were lysed with buffer B with 2.7\u00a0mM KCL. For GFP pulldown after MNase treatment, cells were lysed with buffer B with 75\u00a0mM NaCl, 5\u00a0mM CaCl2, 15\u00a0mM IAA, 3\u00a0mM OPT and 4,000 units of MNase (NEB) per ml of lysate and incubated at 25\u00b0C for 30\u00a0min.Whole\u2010cell lysates were prepared with buffer A: 50\u00a0mM Tris\u2013HCl pH 7.5, 150\u00a0mM NaCl, 5% glycerol, 0.5% NP\u201040, 0.5% sodium deoxycholate, 0.1% SDS, 1\u00a0mM EDTA, 3\u00a0mM 1,10\u2010phenanthroline, and 15\u00a0mM iodoacetamide (IAA), supplemented with phosStop (Roche) and cOmplete mini protease inhibitor (Sigma). Lysates were sonicated and centrifuged at 4\u00b0C for 20\u00a0min at 17,000\u00a0\u00d7\u00a02O2, cells were lysed directly in sample loading buffer and DNA was fragmented by passage through a homogenizer column (Omega Bio\u2010tek). Proteins were subsequently reduced with 10\u00a0mM DTT at 95\u00b0C for 5\u00a0min before loading onto SDS\u2013PAGE gels.For the analysis of Cullin neddylation, cells were lysed directly in urea loading buffer and DNA was fragmented by passage through a homogenizer column (Omega Bio\u2010tek). Proteins were subsequently reduced with 10\u00a0mM DTT at 37\u00b0C for 10\u00a0min before loading onto SDS\u2013PAGE gels. For the analysis of PAR polymer formation after H2O2 continuously for 24\u00a0h. Where indicated, cells were treated with DMSO, olaparib, or DPQ for 1\u00a0h before treatment with H2O2, and continuously thereafter. After 24\u00a0h, cell viability was determined using the CellTiter\u2010Glo assay (Promega). To assess the viability of U2OS cell lines after treatment with camptothecin, cells were plated at 5,000 cells per well of a 96\u2010well plate and 24\u00a0h later were treated with the indicated concentration of camptothecin at 37\u00b0C. After 48\u00a0h, cell viability was measured using the CellTiter\u2010Glo Assay. To analyse cell death after TNF\u03b1\u00a0+\u00a0cycloheximide treatment, U2OS cells were plated at 10,000 cells per well of a 96\u2010well plate. Twenty\u2010four hours after plating, cells were treated with cycloheximide (10\u00a0\u03bcg/ml) for 1\u00a0h and then treated with the indicated amount of TNF\u2010\u03b1. Twenty\u2010four hours after treatment, cell viability was measured using the CellTiter\u2010Glo Assay (Promega).For cell viability analysis of U2OS cell lines, cells were plated at 15,000 cells per well of a 96\u2010well plate and 24\u00a0h later were treated with the indicated concentration of HCell lysates were resolved on Bis\u2010Tris gels (homemade or NuPAGE 4\u201312%) with MOPS buffer (Formedium MOPS\u2010SDS5000) and transferred to nitrocellulose or 0.2\u2010\u03bcm PVDF membranes. Membranes were blocked for 1\u00a0h at RT with 5% milk, or with 5% BSA in TBST, and then incubated with the indicated antibodies overnight at 4\u00b0C. Membranes were then washed 3\u00a0\u00d7\u00a010\u00a0min with TBST and incubated with HRP\u2010conjugated secondary antibodies for 1\u00a0h at RT and then washed 3\u00a0\u00d7\u00a010\u00a0min with TBST. Membranes were incubated with chemiluminescent substrate (EMD Millipore WBKLS0500) and exposed to Konica Blue X\u2010ray film or, for quantification, were imaged using a ChemiDoc Imaging system (Bio\u2010Rad). Images were quantified with ImageJ (NIH).All statistical analysis was performed with GraphPad Prism using the statistical tests indicated in the figure legends.d\u2010lysine (P6407\u20105\u00a0mg Sigma) and 24\u00a0h later were left untreated or treated with 600\u00a0\u03bcM H2O2 for 9\u00a0h at 37\u00b0C. Cells were then washed once with ice\u2010cold phosphate\u2010buffered saline (PBS) and then fixed with ice\u2010cold methanol for 1\u00a0min. Coverslips were washed three times with PBS and then blocked with 3% BSA in PBS for 1\u00a0h at room temperature. Next, coverslips were incubated with primary antibodies diluted in PBS with 3% BSA for 1\u00a0h at room temperature. Coverslips were washed three times with PBS and then incubated with Alexa 488 conjugate anti\u2010rabbit secondary antibody for 1\u00a0h at room temperature. DAPI staining was performed for 5\u00a0min, followed by one wash with PBS. Coverslips were mounted onto slides with ProLong Gold Antifade reagent (Life Technologies) and imaged on a Zeiss LSM 710 confocal microscope. Quantification of nuclear AIF intensity was performed using ImageJ v1.50g .U2OS cells were plated on coverslips coated with 10\u00a0\u03bcg/ml Poly\u2010in\u00a0vitro by incubating NAE (0.15\u00a0\u03bcM), UBE2M/2F (10\u00a0\u03bcM) and NEDD8 (20\u00a0\u03bcM) in reaction buffer for 3\u00a0h at 30\u00b0C. Reactions were stopped by the addition of LDS sample loading buffer.NEDD8 chains were made 2 plates, scraped in ice\u2010cold PBS, collected and centrifuged at 500\u00a0\u00d7\u00a0g for 5\u00a0min. Cell pellets were resuspended in buffer A without SDS, and lysates were spun at 17,000\u00a0\u00d7\u00a0g for 20\u00a0min. The supernatant was collected and incubated with HALO\u2010Link resin (Promega) conjugated to HALO\u2010NEDP1 C163A or DAGC mutant for 3\u00a0h at 4\u00b0C. HALO resin was washed three times with buffer A and three times with 50\u00a0mM Tris\u2013HCl without salt or detergent. Bound proteins were eluted with 100\u00a0\u03bcl of a 50:50 mixture of acetonitrile and 0.1% formic acid for a total of five elutions. Eluate was collected and dried in a speed vac (Thermo Scientific). Protein pellets were resuspended in LDS sample loading buffer and resolved by SDS\u2013PAGE on a 4\u201312% Bis\u2010Tris gel . Gels were stained with InstantBlue colloidal Coomassie (Expedeon). Bands were then excised, washed with water, dehydrated in acetonitrile and rehydrated in 50\u00a0mM Tris\u2013HCl pH 8.0. Gel slices were alkylated with 20\u00a0mM chloroacetamide, dehydrated in acetonitrile and then transferred to 50\u00a0mM triethylammonium bicarbonate. Gel bands were incubated with trypsin (5\u00a0\u03bcg/ml) overnight at 30\u00b0C. Peptides were extracted with acetonitrile, gel bands were incubated in 0.1% trifluoroacetic acid (TFA), and then, peptides were extracted two more times with acetonitrile. Extracted peptides were dried in a speed vac and then resuspended in 0.1% TFA/water. Samples were analysed on a LTQ Orbitrap Velos Pro mass spectrometer (Thermo Fisher Scientific) coupled to an Ultimate 3000 UHPLC system with 15\u2010cm Acclaim PepMap 100 analytical column with an addition Pepmap trapping column . Acquisition settings were lockmass of 445.120024, MS1 with 60,000 resolution, top 20 CID MS/MS using rapid scan, monoisotopic precursor selection, unassigned charge states and z\u00a0=\u00a01 rejected, dynamic exclusion of 60s with repeat count of 1. One\u2010hour linear gradients were performed from 5% solvent B to 35% solvent B . Raw files were processed in Proteome Discoverer 2.0 (Thermo Scientific), with Mascot 2.4.1 (Matrix Science),and subsequently processed in Scaffold 4.4.6 (Proteome Software) Searches were performed with a peptide tolerance of 10\u00a0ppm (monoisotopic) and a fragment tolerance of 0.60\u00a0Da (monoisotopic) or with MaxQuant v1.5.7.4 for label\u2010free quantitative and iBAQ analysis. Settings were fixed modifications of carbamidomethyl (C), variable modifications of oxidation (M), dioxidation (M), LRGG (K) and GlyGly (K). Protein identifications were filtered with a 1% FDR. Raw files were re\u2010searched separately with variable modification for acetylated peptides acetyl (K) or phosphorylated peptides phospho (STY); both searches included the fixed modifications of carbamidomethyl (C), variable modifications of oxidation (M) and dioxidation (M).For HALO pulldown for mass spectrometry analysis, NEDP1 KO HEK 293 cells were grown on 150\u2010cmin\u00a0vitro neddylation reactions, the reactions were resolved on a 4\u201312% Bis\u2010Tris gel , stained with InstantBlue colloidal Coomassie (Expedeon), and processed for in\u2010gel trypsin digest, as described above.For analysis of g for 5\u00a0min. Cell pellets were washed two times with 10\u00a0ml of PBS and then lysed with 8\u00a0M urea and 0.5% CHAPS. Protein concentration was determined by BCA assay using the manufacturer's protocol . To remove salt and impurities, protein was precipitated with acetone. Four times the sample volume of \u221220\u00b0C acetone was added and the solution was incubated at \u221220 for 2\u00a0h. The sample was then centrifuged at 4\u00b0C for 20\u00a0min at 17,000\u00a0\u00d7\u00a0g. The supernatant was decanted, and the pellet was washed with four volumes of 80% acetone, followed by centrifugation at 4\u00b0C for 5\u00a0min at 17,000\u00a0\u00d7\u00a0g. The supernatant was decanted, the pellet was briefly centrifuged again, and residual acetone was removed. The pellet was dried at room temperature for 5\u00a0min. Pellets were resuspended with rehydration buffer . 65\u00a0mM DTT and trace bromophenol blue) to 30\u00a0\u03bcg/ml. Subsequently, 90\u00a0\u03bcg of protein was added to a 7\u00a0cm, pH 5\u20138 IPG strip (Bio\u2010Rad) and allowed to rehydrate on the strip for 14\u00a0h, followed by focusing for 12\u00a0h for a total of 22\u00a0kVhrs. For two\u2010dimensional electrophoresis, separation strips were equilibrated with equilibration buffer (Bio\u2010Rad) with 50\u00a0mM DTT for 15\u00a0min. The buffer was replaced and the strips equilibrated again for 15 additional minutes. IPG strips were loaded onto precast 4\u201020% TGX gels (Bio\u2010Rad) and processed for Western blot analysis.To prepare lysate from wild\u2010type U2OS cells, one T\u2010150 flask was harvested with 5\u00a0ml trypsin\u2010EDTA (Life Technologies). Detached cells were collected with fresh media and centrifuged at 500\u00a0\u00d7\u00a0For 2D gel electrophoresis analysis of GST pulldowns, USOS NEDP1 KO cells were harvested with lysis buffer B, as described above for the endogenous PARP\u20101 immunoprecipitation. Next, 20\u00a0mg of lysate was used for each pulldown with 100\u00a0\u03bcg GST or 100\u00a0\u03bcg GST\u2010PARP\u20101 Zn1\u00a0+\u00a02 preconjugated to Sepharose 4B beads. The beads and lysates were incubated at 4\u00b0C for 2\u00a0h with rotation. The beads were then washed four times with wash buffer , followed by three washes with salt\u2010free buffer (50\u00a0mM Tris\u2013HCl pH 7.5). Bound proteins were eluted with 8\u00a0M urea and 0.5% CHAPS and were processed for isoelectric focusing, as described above.MJK performed most experiments. RH designed NEDP1 constructs, performed HALO\u2010NEDP1 pulldowns and sample preparation for the HEK 293 mass spectrometry experiment and performed some cell viability experiments. OH performed microscopy of AIF translocation. RG processed mass spectrometry samples and performed database searches. RB performed 2D gel electrophoresis. MJK, RG, RH, MT and TK analysed mass spectrometry results. MJK, RH and TK designed experiments. MJK and TK wrote the manuscript with input from all authors. RH and TK conceived the project, and TK supervised the work.The authors declare that they have no conflict of interest.Expanded View Figures PDFClick here for additional data file.Table\u00a0EV1Click here for additional data file.Review Process FileClick here for additional data file."} +{"text": "L\u2032)FeCp*(CO)][PF6] [L\u2032 = S\u2014C(Me)=C(Me)\u2014S\u2014(CH2)3\u2014S\u2014C(Me)=C(Me)\u2014S] is reported.The new structural NiFe hydrogenase model PF6 or [Ni(L\u2032)FeCp*(CO)]PF6, is composed of the nickel complex fragment [Ni(L\u2032)] coordinated as a metalloligand (using S1 and S4) to the [FeCp*(CO)]+ fragment, where (L\u2032)2\u2212 is [S\u2014C(Me)=C(Me)\u2014S\u2014(CH2)3\u2014S\u2014C(Me)=C(Me)\u2014S]2\u2212 and where Cp*\u2212 is cyclo-C5(Me)5\u2212 (penta\u00admethyl\u00adcyclo\u00adpenta\u00addien\u00adyl). The ratio of hexa\u00adfluoro\u00adphosphate anion per complex cation is 1:1. The structure at 150\u2005K has ortho\u00adrhom\u00adbic (Pbcn) symmetry. The atoms of the complex cation are located on general positions (multiplicity = 8), whereas there are two independent hexa\u00adfluoro\u00adphosphate anions, each located on a twofold axis . The structure of the new dimetallic cation [Ni(L\u2032)FeCp*(CO)]+ can be described as containing a three-legged piano-stool environment for iron [Cp*Fe(CO)\u2018S2\u2019] and an approximately square-planar \u2018S4\u2019 environment for Ni. The NiS2Fe diamond-shaped substructure is notably folded at the S\u2014S hinge: the angle between the NiS2 plane and the FeS2 plane normals is 64.85\u2005(6)\u00b0. Largely because of this fold, the nickel\u2013iron distance is relatively short, at 2.9195\u2005(8)\u2005\u00c5. The structural data for the complex cation, which contains a new unsaturated \u2018S4\u2019 ligand (two C=C double bonds), provide an inter\u00adesting comparison with the known NiFe hydrogenase models containing a saturated \u2018S4\u2019-ligand analogue having the same number of carbon atoms in the ligand backbone, namely with the structures of [Ni(L)FeCp(CO)]+ and [Ni(L)FeCp*(CO)]+ , where (L)2\u2212 is [S\u2014CH2\u2014CH2\u2014S\u2014(CH2)3\u2014S\u2014CH2\u2014CH2\u2014S]2\u2212 and Cp\u2212 is cyclo\u00adpenta\u00addienyl. The saturated analogues [Ni(L)FeCp(CO)]+ and [Ni(L)FeCp*(CO)]+ have similar Ni\u2014Fe distances: 3.1727\u2005(6), 3.1529\u2005(7)\u2005\u00c5 (two independent mol\u00adecules in the unit cell) and 3.111\u2005(5)\u2005\u00c5, respectively, for the two complexes, whereas [Ni(L\u2032)FeCp*(CO)]+ described here stands out with a much shorter Ni\u2014Fe distance [2.9196\u2005(8)\u2005\u00c5]. Also, [Ni(L)FeCp(CO)]+ and [Ni(L)FeCp*(CO)]+ show inter\u00adplanar fold angles that are similar between the two: 39.56\u2005(5), 41.99\u2005(5) (independent mol\u00adecules in the unit cell) and 47.22\u2005(9) \u00b0, respectively, whereas [Ni(L\u2032)FeCp*(CO)]+ possesses a much more pronounced fold [64.85\u2005(6)\u00b0]. Given that larger fold angles and shorter Ni\u2014Fe distances are considered to be structurally closer to the enzyme, unsaturation in an \u2018S4\u2019-ligand of the type (S\u2014C2\u2014S\u2014C3\u2014S\u2014C2\u2014S)2\u2212 seems to increase structural resemblance to the enzyme for structural models of the type [Ni(\u2018S4\u2019)FeCpR(CO)]+ (CpR = Cp or Cp*).The complex cation in the title compound, (carbonyl-1\u03ba The first \u2018S4\u2019 ligand used in this capacity featured a saturated two\u2013three\u2013two carbon linker, in L2\u2212 = [S\u2014CH2\u2014CH2\u2014S\u2014(CH2)3\u2014S\u2014CH2\u2014CH2\u2014S]2\u2212 FeCpR(CO)]+ model based on an analogous but unsaturated \u2018S4\u2019 ligand, namely L\u20322\u2212 = [S\u2014C(Me)=C(Me)\u2014S\u2014(CH2)3\u2014S\u2014C(Me)=C(Me)\u2014S]2\u2212 Fe(CpR)(CO)]+ complex in which the \u2018S4\u2019 ligand has a four-carbon linker in the remote portion of the backbone FeCp*(CO)]+ was obtained as solvent-free crystals containing the PF6\u2212 counter-ion. A drawing showing both cation and anion in this salt is shown below (see Supramol\u00adecular features), and the intra\u00admolecular structural features of the cation are discussed first. The structure of [Ni(L\u2032)FeCp*(CO)]+ is shown in Fig.\u00a034\u2019 environment for Ni (sum of bond angles around Ni1 = 359.83\u00b0). Selected metal\u2013ligand distances are Ni1\u2014S1 = 2.1616\u2005(11), Ni1\u2014S2 = 2.1530\u2005(12), Ni1\u2014S3 = 2.1507\u2005(11), Ni1\u2014S4 = 2.1563\u2005(12)\u2005\u00c5, and Fe1\u2014S1 = 2.3309\u2005(12), Fe1\u2014S4 = 2.3602\u2005(12), Fe1\u2014C11 = 1.768\u2005(5), Fe1\u2014C1 = 2.080\u2005(4), Fe1\u2014C2 = 2.107\u2005(4), Fe1\u2014C3 = 2.126\u2005(4), Fe1\u2014C4 = 2.138\u2005(4), Fe1\u2014C5 = 2.098\u2005(4)\u2005\u00c5. The inter\u00admetallic (Ni1\u2014Fe1) distance is relatively short, i.e. 2.9195\u2005(8)\u2005\u00c5. The NiS2Fe diamond is markedly folded at the S\u2014S hinge: the angle between the NiS2 plane and the FeS2 plane normals is 64.85\u2005(6)\u00b0, and this fold largely accounts for the short nickel\u2013iron distance.[Ni(L\u20322\u2212 with those of literature complexes using the saturated ligand L2\u2212. The structures of [Ni(L)FeCp(CO)]+, as the PF6\u2212 salt/ CH2Cl2 solvate FeCp*(CO)]+, as the PF6\u2212 salt FeCp(CO)]+ and [Ni(L)FeCp*(CO)]+ show Ni\u2014Fe distances that are similar for the two, 3.1727\u2005(6)/3.1529\u2005(7)\u2005\u00c5 (two independent mol\u00adecules in the unit cell) and 3.111\u2005(5)\u2005\u00c5, respectively, for the two complexes. The [Ni(L\u2032)FeCp*(CO)]+ complex, on the other hand, has a much shorter Ni\u2014Fe distance . Also, [Ni(L)FeCp(CO)]+ and [Ni(L)FeCp*(CO)]+ show inter\u00adplanar fold angles that are similar for the two, 39.56\u2005(5)/41.99\u2005(5)\u00b0 (two independent mol\u00adecules in the unit cell) and 47.22\u2005(9)\u00b0, respectively, while [Ni(L\u2032)FeCp*(CO)]+ has a much larger fold angle of 64.85\u2005(6)\u00b0 (see above). The large fold angle and short Ni\u2014Fe distance observed in the complex with the unsaturated ligand L\u2032 match the structure of the enzymatic active site more closely than the angles/distances of the complexes containing the saturated ligand L. For eight structurally characterized enzymes, the dihedral angles range from 59 to 99\u00b0 and the Ni\u2014Fe distances range from 2.53 to 2.97\u2005\u00c5 2\u2212 can increase structural resemblance to the enzyme in models of the type [Ni(\u2018S4\u2019)FeCpR(CO)]+. Structural similarity to the enzyme in models was, in alternative approaches, also favoured when additional donor atoms were incorporated into the ligand chain (such as \u2018S3N2\u2019) or where two bidentate chelate ligands were used instead of one large \u2018S4\u2019 ligand. FeCp*(CO)]+ model with four carbon atoms, instead of three, in the remote portion of the backbone \u2005\u00c5 and 62.48\u2005(4)\u00b0, respectively FeCp*(CO)]+ was shown to be active as a hydrogen-production catalyst Cp*(CO)]+ complex, with the unsat\u00adurated \u2018S4\u2019 ligand L\u2032, might warrant deeper investigation. We conclude that the introduction of unsaturation in the \u2018S4\u2019 ligand led to a better structural model relative to the unsaturated ligand, highlighting a new variant of the classic [Ni(\u2019S4\u2019)FeCpR(CO)]+-type hydrogenase model.In the following discussion, we compare the structural features obtained with the unsaturated ligand L\u2032)FeCp*(CO)]+ with hexa\u00adfluoro\u00adphosphate anions, without solvent mol\u00adecules and without any solvent-accessible void. The ratio of hexa\u00adfluoro\u00adphosphate anion per complex cation is 1:1. The atoms of the complex cation are situated on general positions (multiplicity = 8), whereas there are two independent hexa\u00adfluoro\u00adphosphate anions, each situated on a twofold axis . A picture of the packing is shown in Fig.\u00a04B\u22efF4 = 2.55\u2005\u00c5; C15\u2014H15B\u22efF3i = 2.55\u2005\u00c5; C21\u2014H21C\u22efF4ii = 2.48\u2005\u00c5; C22\u2014H22C\u22efF1iii = 2.52\u2005\u00c5) and a C\u2014H\u22efO short contact (C14\u2014H14A\u22efO1 = 2.41\u2005\u00c5) .The structure results from packing of discrete cations +, they are not discussed further. LAZVUE FeCp(CO)]+ , MUDXOA FeCp*(CO)]+ , and SUWWAJ FeCp*(CO)]+ . These three complex cations are discussed in detail above.The Cambridge Crystallographic Database .The syntheses were performed in dried solvents under an inert atmosphere using standard glove-box (MBraun) and Schlenk techniques. Deuterated NMR solvents were from Cambridge Isotopes. using 1,3-di\u00adbromo\u00adpropane. Na2[Ni(S2C2Me2)] was prepared from Ni(S2C2Me2)2 by reduction with excess sodium in THF , until the colour had changed from deep purple to brown\u2013yellow. The subsequent alkyl\u00adation of [Ni(S2C2Me2)]2\u2212 using 1,3-di\u00adbromo\u00adpropane was performed analogously to the procedure described by Schrauzer and co-workers for the closely related Ni(S\u2014C(Ph)=C(Ph)\u2014S\u2014(CH2)3\u2014S\u2014C(Ph)=C(Ph)\u2014S). ][PF6]: This precursor for the iron part of the complex was prepared according to the general procedure for [Cp*Fe(CO)2(solvent)]+ given by Catheline & Astruc (1984[PF6]: Crude [Cp*Fe(CO)2(NCMe)][PF6] was combined with 6\u2005ml of aceto\u00adnitrile and filtered through a glass filter frit. While purging with argon, the reaction was irradiated with UV\u2013visible light for 16\u2005h. Under an inert atmosphere, a solution of 155\u2005mg (0.46\u2005mmol) of Ni(L\u2032) in ca 7\u2005ml of di\u00adchloro\u00admethane was added. The reaction mixture was heated under active argon flow to 325\u2005K for 2\u2005h. After cooling to room temperature, the volatiles were slowly removed under vacuum. The solid was dried under vacuum and stored in the glove-box. Yield of crude product: 253\u2005mg (75%). 1H NMR \u03b4 1.60 ; \u03b4 1.91 ; \u03b4 1.96 ; \u03b4 2.31 ; \u03b4 2.0\u20133.7 . Note that the sample thus prepared showed a 1H NMR signal for metal-coordinated aceto\u00adnitrile. The purpose of the prolonged photolysis was to remove all CO from iron, in order to selectively prepare [Ni(L\u2032)FeCp*(NCMe)][PF6]. However, the sample obtained appeared to be a mixture of [Ni(L\u2032)FeCp*(CO)][PF6] and [Ni(L\u2032)FeCp*(NCMe)][PF6] and is thus referred to as [Ni(L\u2032)FeCp*(CO/NCMe)][PF6]. Yet, crystallization from acetone yielded exclusively [Ni(L\u2032)FeCp*(CO)][PF6], in crystalline form.[Ni(L\u2032)FeCp*(CO)][PF6]: 11\u2005mg of [Ni(L\u2032)FeCp*(CO/NCMe)][PF6] were dissolved in 1.5\u2005ml of acetone and filtered through 1\u2005cm of Celite. Through solvent vapor diffusion, by placing the loosely capped vial into a larger vessel containing diethyl ether vapour (and some liquid), crystals of [Ni(L\u2032)FeCp*(CO)][PF6] were grown within two days at 308\u2005K.Crystallization of [Ni(Uiso(H) = 1.2Ueq(C) or 1.5Ueq(Cmeth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2056989018010939/zl2735sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018010939/zl2735Isup2.hklStructure factors: contains datablock(s) I. DOI: 1859284CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the mol\u00adecules are linked via O\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds, forming chains along [101].The title mol\u00adecule is comprised of two non-coplanar benzene rings connected by an imino group in a 16H17NO4, the dihedral angle between benzene rings is 72.7\u2005(2)\u00b0. The meth\u00adoxy groups are rotated by 2.4\u2005(2) and \u22124.9\u2005(2) (benzil\u00adidene moiety) and by 5.6\u2005(3)\u00b0 (aniline moiety) relative to the adjacent benzene ring. In the crystal, the mol\u00adecules are linked into chains along [101] through C\u2014H\u22efO and O\u2014H\u22efN hydrogen bonds.In the title compound, C The meth\u00adoxy groups are almost co-planar with the planes of the adjacent aromatic rings .The mol\u00adecular structure of the title molecule is shown on Fig.\u00a01via C7\u2014H7\u22efO2ii and O2\u2014H2\u22efN1i hydrogen bonding \u00admeth\u00adyl]phenol, (I)et al., 1992E)-5-meth\u00adoxy-2-[(4-meth\u00adoxy\u00adphenyl\u00adimino)\u00admeth\u00adyl]phenol, (II) \u00admeth\u00adyl]phenol, (IV) and 50\u2005ml of distilled water. The reaction mixture was taken in a clean 250\u2005ml round-bottom flask and stirred well with a magnetic stirrer. It was then refluxed for 7\u2005h. The dark-yellow product that formed was separated by filtration, dried under vacuum and recrystallized from methanol solution upon slow evaporation for two days .4-Hy\u00addroxy-3,5-di\u00admeth\u00adoxy\u00adbenzaldehyde (0.05\u2005mol) was added to a mixture of 50\u2005ml of methanol and Uiso(H) = 1.2Ueq(C) or 1.5Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018013713/ld2146sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018013713/ld2146Isup3.cmlSupporting information file. DOI: 10.1107/S2056989018013713/ld2146Isup3.hklStructure factors: contains datablock(s) I. DOI: 1843910CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the original publication, values of the doses of insulin glargine, the most commonly used basal insulin analogue under the \u2018Discussion\u2019 section was incorrectly published.The sentence \u201c\u2026was similar (3100.314 \u00b1 110.1298 U/kg body weight/day at baseline) to that in the Swedish study (0.33 \u00b1 0.16 U/kg body weight/day)\u201d.Should actually say \u201c\u2026was similar (0.31 \u00b1 0.12 U/kg body weight/day at baseline) to that in the Swedish study (0.33 \u00b1 0.16 U/kg body weight/day)\u201d."} +{"text": "The asymmetric unit of the title compound consists of two independent mol\u00adecules differing slightly in the conformations of the seven-membered rings and the butyl substituents. 16H20N2O2, consists of two independent mol\u00adecules differing slightly in the conformations of the seven-membered rings and the butyl substituents, where the benzene rings are oriented at a dihedral angle of 34.56\u2005(3)\u00b0. In the crystal, pairwise inter\u00admolecular C\u2014H\u22efO and complementary intra\u00admolecular C\u2014H\u22efO hydrogen bonds form twisted strips extending parallel to (012). These strips are connected into layers parallel to (111) by additional inter\u00admolecular C\u2014H\u22efO hydrogen bonds. The layers are further joined by C\u2014H\u22ef\u03c0 inter\u00adactions. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (65.5%), H\u22efC/C\u22efH (16.0%) and H\u22efO/O\u22efH (15.8%) inter\u00adactions.The asymmetric unit of the title compound, C Rings B and D have boat conformations with slightly different Cremer\u2013Pople puckering parameters . In the n-butyl substituents, the C13\u2014C14\u2014C15\u2014C16 [177.96\u2005(13)\u00b0] and C29\u2014C30\u2014C31\u2014C32 [174.97\u2005(12)\u00b0] chains also have slightly different torsion angles. The conformation of the 2-oxo\u00adpropyl\u00adidene units are partially determined by the intra\u00admolecular N\u2014H\u22efO hydrogen bonds (Table\u00a01The asymmetric unit of the title compound consists of two independent mol\u00adecules differing modestly in the conformations of the seven-membered s Table\u00a01 The r.m.Bnz\u22efOOxoprp (Bnz = Benzene and Oxoprp = 2-oxo\u00adpropyl\u00adidene) and complementary intra\u00admolecular C\u2014HBnz\u22efOBnzdzp hydrogen bonds (Table\u00a01Bnz\u22efOOxoprp and C\u2014HBnzdzp\u22efOBnzdzp hydrogen bonds (Table\u00a01Bnzdzp\u22ef\u03c0 and C\u2014HBty\u22ef\u03c0 (Bty = n-but\u00adyl) inter\u00adactions analysis contacts -4-(2-oxo\u00adpropyl\u00adidene)-4,5-di\u00adhydro-1H-benzo[b]diazepin-2(3H)-one (2.38\u2005mmol) in 15\u2005ml of di\u00adchloro\u00admethane were added 1.5 eq of 1-bromo\u00adbutane, (3.57\u2005mmol) of potassium hydroxide dissolved in water and 0.23\u2005mmol of tetra-n-butyl ammonium bromide (BTBA). The mixture was kept under magnetic stirring at room temperature for 48\u2005h. A little water was added and then the organic phase was extracted. The mixture obtained was chromatographed on a column of silica gel (eluent hexa\u00adne/ethyl acetate 8/2) to give three products. The title compound was isolated as the major product in a yield of 77%.To a solution of (Uiso(H) = 1.5Ueq(C). The remaining H atoms were located in a difference-Fourier map and were freely refined. The crystal studied was twinned.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018014779/xu5946sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018014779/xu5946Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018014779/xu5946Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018014779/xu5946Isup4.cmlSupporting information file. DOI: 1874203CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II and AuI cations to form a one-dimensional polymeric compound with free di\u00adcyano\u00adaurate anions.Cyanide anions bridge Fe 2(C10H8N2)2(H2O)2][Au(CN)2]}n, the FeII ion, which is located on a twofold rotation axis, has a slightly distorted FeN4O2 octa\u00adhedral geometry. It is coordinated by two phenyl\u00adpyrazine mol\u00adecules, two water mol\u00adecules and two di\u00adcyano\u00adaurate anions, the Au atom also being located on a second twofold rotation axis. In the crystal, the coordinated di\u00adcyano\u00adaurate anions bridge the FeII ions to form polymeric chains propagating along the b-axis direction. In the crystal, the chains are linked by Owater\u2014H\u22efNdi\u00adcyano\u00adaurate anions hydrogen bonds and aurophillic inter\u00adactions [Au\u22efAu = 3.5661\u2005(3)\u2005\u00c5], forming layers parallel to the bc plane. The layers are linked by offset \u03c0\u2013\u03c0 stacking inter\u00adactions [inter\u00adcentroid distance = 3.643\u2005(3)\u2005\u00c5], forming a supra\u00admolecular metal\u2013organic framework.In the title polymeric complex, {[Fe(CN) Applying external stimuli, such as temperature, pressure, magnetic field, light irradiation or adding a guest can affect this kind of the compound and change their properties significantly , which consist of metal ions and organic ligand linkers, are studied intensively. Fe-based coordination polymers with N-donor bridging ligands are well known as compounds with switchable spin states \u2005\u00c5] in axial positions, and two N atoms of cyano bridges and two water O atoms of water mol\u00adecules [Fe1\u2014O1 = 2.122\u2005(4)\u2005\u00c5] in equatorial positions. The two CN\u2212 anions bridge the FeII and AuI cations [Fe1\u22efAu1 = 5.244\u2005(3)\u2005\u00c5] to form a one-dimensional polymeric structure with bond lengths Fe1\u2014N1 = 2.107\u2005(5)\u2005\u00c5 and Fe1\u2014\u2013N2 = 2.117\u2005(6)\u2005\u00c5 is \u03a3|90 - \u0398| = 8.53\u00b0, where \u0398 are the cis-N\u2014Fe\u2014O and cis-N\u2014Fe\u2014N angles in the coordination environment of the FeII atom.The structure of the title compound features a one-dimensional chain motif that runs parallel to the crystallographic s Figs. 1 and 2 \u25b8.water\u2014H\u22efN hydrogen bonds .The crystal packing features different types of weak inter\u00adactions in water (2.5\u2005ml), the second was a mixture of water/aceto\u00adnitrile and the third layer was a solution of 2-phenyl\u00adpyrazine and [Fe(OTs)2]\u00b76H2O (OTs = p-toluene\u00adsulfonate) in aceto\u00adnitrile (2.5\u2005ml) with 0.3\u2005ml of water. After two weeks, yellow crystals grew in the second layer; these were collected and maintained under the mother solution until measured.Crystals of the title compound were prepared by the slow diffusion method between three layers in a 10\u2005ml tube. The first layer was a solution of K[Au(CN)Uiso(H) = 1.2Uiso(C). The idealized OH2 group was fixed using an AFIX 7 command that allowed the H atoms to ride on the O atom and rotate around the bond.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019009678/su5503sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989019009678/su5503Isup2.hklStructure factors: contains datablock(s) I. DOI: 1938914CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "C\u2014H\u22efO hydrogen bonds, weak C\u2014H\u22ef\u03c0 and weak offset \u03c0\u2013\u03c0 stacking inter\u00adactions stabilize the packing.In the title pyridazinone derivative, the unsubstituted phenyl ring and the pyridazine ring are inclined to each other, making a dihedral angle of 17.41\u2005(13)\u00b0, whereas the Cl-substituted phenyl ring is nearly orthogonal to the pyridazine ring [88.19\u2005(13)\u00b0], C 21H19ClN2O3, is not planar. The unsubstituted phenyl ring and the pyridazine ring are inclined to each other, making a dihedral angle of 17.41\u2005(13)\u00b0 whereas the Cl-substituted phenyl ring is nearly orthogonal to the pyridazine ring [88.19\u2005(13)\u00b0]. In the crystal, C\u2014H\u22efO hydrogen bonds generate dimers with R22(10) and R22(24) ring motifs which are linked by C\u2014H\u22efO inter\u00adactions, forming chains extending parallel to the c-axis direction. The inter\u00admolecular inter\u00adactions were investigated using Hirshfeld surface analysis and two-dimensional fingerprint plots, revealing that the most significant contributions to the crystal packing are from H\u22efH (44.5%), C\u22efH/H\u22efC (18.5%), H\u22efO/H\u22efO (15.6%), Cl\u22efH/H\u22efCl (10.6%) and C\u22efC (2.8%) contacts.The title pyridazinone derivative, C A weak n Table\u00a01. Weak aret al., 2016viz. 4-benzyl-6-p-tolyl\u00adpyridazin-3(2H)-one to 1.1730 (blue) a.u. The three-dimensional dnorm surface of the title mol\u00adecule is illustrated in Fig.\u00a03a. The pale-red spots symbolize short contacts and negative dnorm values on the surface and correspond to the C\u2014H\u22efO inter\u00adactions . The convex blue regions symbolize hydrogen-donor groups and the concave red regions hydrogen-acceptor groups. \u03c0\u2013\u03c0 inter\u00adactions are generally indicated by adjacent red and blue triangles in the shape-index map, as is the case for the title mol\u00adecule.The shape-index map of the title mol\u00adecule was generated in the range \u22121 to 1\u2005\u00c5 Fig.\u00a03b. The cc). The curvedness plot of the title complex shows large regions of green with a relatively flat (i.e. planar) surface area, indicating the presence of \u03c0\u2013\u03c0 stacking inter\u00adactions, while the blue regions demonstrate areas of curvature.The curvedness map of the title complex was generated in the range \u22124.0 to 0.4\u2005\u00c5 Fig.\u00a03c. The ca, delineated into H\u22efH, H\u22efC/ C\u22efH, H\u22efO/O\u22efH, H\u22efCl/Cl\u22efH, C\u22efC contacts associated with their relative contributions to the Hirshfeld surface in Fig.\u00a04b\u2013f, respectively. The most important inter\u00admolecular inter\u00adaction is H\u22efH, contributing 44.5% to the overall crystal packing, with the centre of the peak de = di = 1.18\u2005\u00c5 . H\u22efC/ C\u22efH contacts, with a 18.5% contribution to the Hirshfeld surface, indicate the presence of the weak C\u2014H\u22ef\u03c0 inter\u00adaction . H\u22efO/O\u22efH contacts arising from inter\u00admolecular C\u2014H\u22efO hydrogen bonding make a 15.6% contribution to the Hirshfeld surface and are represented by a pair of sharp spikes in the region de + di \u223c2.35\u2005\u00c5 The C\u22efC contacts are a measure of \u03c0\u2013\\p stacking inter\u00adactions and contribute 2.8% of the Hirshfeld surface. They appear as an arrow-shaped distribution at de + di \u223c3.3\u2005\u00c5. Another contact to the Hirshfeld surface is from H\u22efCl/Cl\u22efH inter\u00adactions (10.6%).The overall two-dimensional fingerprint plot is illustrated in Fig.\u00a04\u2005\u00c5 Fig.\u00a04b. H\u22efC/ n Table\u00a01. Two paint Fig.\u00a04c. H\u22efO/OH)-one in 30\u2005ml of tetra\u00adhydro\u00adfuran (THF), potassium carbonate was added. The mixture was refluxed for 1\u2005h. After cooling, ethyl bromo\u00adacetate was added and the mixture was refluxed for 8\u2005h. The precipitated material was removed by filtration and the solvent evaporated under vacuum. The residue was purified through silica gel column chromatography using hexa\u00adne/ethyl acetate (4:6 v/v). Slow evaporation at room temperature led to formation of single crystals with a yield of 70%.To a solution of 4-(3-di\u00adchloro\u00adbenz\u00adyl)-6-phenyl\u00adpyridazin-3(2Uiso(H) = 1.2Ueq(C)], C\u2014H = 0.96\u2005\u00c5 for methyl\u00adene [Uiso(H) = 1.5Ueq(C)], C\u2014H = 0.93\u2005\u00c5 for aromatic [Uiso(H) = 1.2Ueq(C)] and C\u2014H = 0.98\u2005\u00c5 for methine [Uiso(H) = 1.2Ueq(C)] H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019007424/wm5505sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019007424/wm5505Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019007424/wm5505Isup3.cmlSupporting information file. DOI: 1917654CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure of the title compound, the mol\u00adecules are linked by pairs of O\u2014H\u22efO hydrogen bonds, forming inversion dimers with an 15H15NO2, is a Schiff base that exists in the keto\u2013enamine tautomeric form and adopts a Z configuration. The mol\u00adecule is almost planar, with the two phenyl rings twisted relative to each other by 9.60\u2005(18)\u00b0. There is an intra\u00admolecular N\u2014H\u22efO hydrogen bond present forming an S(6) ring motif. In the crystal, pairs of O\u2014H\u22efO hydrogen bonds link adjacent mol\u00adecules into inversion dimers with an R22(18) ring motif. The dimers are linked by very weak \u03c0\u2013\u03c0 inter\u00adactions, forming layers parallel to (The title compound, C RCH=N\u2013R\u2032) and are prepared by condensation reactions between amines and active carbonyl compounds -6-{[(2-hy\u00addroxy-4-methyl\u00adphen\u00adyl)amino]\u00admethyl\u00adidene}-4-methyl\u00adcyclo\u00adhexa-2,4-dien-1-one, was obtained in crystalline form from the reaction of 2-amino-5-methyl\u00adphenol with 2-hy\u00addroxy-5-methyl\u00adbenz\u00adaldehyde. We report here its synthesis conditions and the mol\u00adecular and crystal structures, supplemented by Hirshfeld surface analysis.In the current study, a new Schiff base, . The mol\u00adecule is almost planar, with an r.m.s. deviation of 0.1061\u2005\u00c5 for the complete mol\u00adecule except the H atoms [largest deviation 0.176\u2005(3)\u2005\u00c5 for C8]. The two phenyl rings (C1\u2013C6 and C9\u2013C14) are inclined by 9.60\u2005(18)\u00b0. The C1\u2014O1 bond length [1.356\u2005(3)\u2005\u00c5] to the hy\u00addroxy group is in the normal range, while the C14=O2 bond length is comparatively elongated [1.302\u2005(4)\u2005\u00c5] due to the involvement of the carbonyl O atom in an intra\u00admolecular N\u2014H\u22efO hydrogen bond, forming an S(6) ring motif. The C6\u2014N1 and C8=N1 bond lengths are 1.404\u2005(4) and 1.310\u2005(4)\u2005\u00c5, respectively. Overall, the bond lengths in the title structure compare well with those of other keto\u2013enamine tautomers known from the literature (see: Database Survey).The mol\u00adecular structure of the title compound is illustrated in Fig.\u00a01Cg1\u22efCg2 = 4.721\u2005(2)\u2005\u00c5; Cg1 and Cg2 are the centroids of the C1\u2013C6 and C9\u2013C14 rings, respectively], forming layers parallel to -2-[(2-hy\u00addroxy\u00adphenyl\u00adiminio)meth\u00adyl]phenolate fragment revealed 25 hits where this fragment adopts the keto\u2013enamine tautomeric form. Nearly all bond lengths in the title structure are the same within standard uncertainties as the corresponding bond lengths in the structures of 2,4-di\u00adchloro-6-{[(2-meth\u00adoxy\u00adphen\u00adyl)iminio]meth\u00adyl}phenolate hydrate -[(2-hy\u00addroxy\u00adphen\u00adyl)iminio]meth\u00adyl}-4-methyl\u00adphenolate -4-hy\u00addroxy-2-[(2-hy\u00addroxy\u00adphen\u00adyl)iminiometh\u00adyl]phenolate -[(2-hy\u00addroxy-5-methyl\u00adphen\u00adyl)iminio]meth\u00adyl}-4-(tri\u00adfluoro\u00admeth\u00adoxy)phenolate surface resolution. Fig.\u00a04b shows the mol\u00adecular electrostatic potential plotted over the three-dimensional Hirshfeld surface using the STO-3G basis set in the range \u22120.0975 to 0.2197 a.u. within the Hartree\u2013Fock level of theory. The O\u2014H\u22efO hydrogen-bond donors and acceptors are shown as blue and red areas around the atoms related with positive (hydrogen-bond donors) and negative (hydrogen-bond acceptors) electrostatic potentials, respectively.A Hirshfeld surface analysis points related to H\u22efH contacts that represent a 55.2% contribution in the title structure. In Fig.\u00a05c, two symmetrical wings on the left and right sides indicate C\u22efH/H\u22efC inter\u00adactions with a contribution of 22.3%. Furthermore, there are O\u22efH/H\u22efO , C\u22efC (4.9%) and C\u22efN/N\u22efC (2.6%) contacts.Fig.\u00a05Fig.\u00a06The title compound was prepared by refluxing a mixture of 2-hy\u00addroxy-5-methyl\u00adbenzaldehyde in ethanol (15\u2005ml) and 2-amino-5-methyl\u00adphenol in ethanol (15\u2005ml) for 5\u2005h. Single crystals of the title compound for X-ray analysis were obtained by slow evaporation of an ethanol solution .Uiso(H) = 1.5Ueq. The C-bound H atoms were positioned geometrically and refined using a riding model with C\u2014H = 0.93 and Uiso(H) = 1.2Ueq(C) for aromatic H atoms, and with C\u2014H = 0.96\u2005\u00c5 and Uiso(H) = 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019006583/wm5504sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019006583/wm5504Isup2.hklStructure factors: contains datablock(s) I. DOI: 1902148CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "BM) grafts in F1 hybrid recipients is due to natural killer (NK) cell\u2013mediated rejection triggered through \u201cmissing self\u201d recognition. \u201cHybrid resistance\u201d has usually been investigated in lethally irradiated F1 recipients in conjunction with pharmacological activation of NK cells. Here, we investigated BM\u2010directed NK\u2010cell alloreactivity in settings of reduced conditioning. Nonlethally irradiated (1\u20103\u00a0Gy) or nonirradiated F1 (C57BL6\u00a0\u00d7\u00a0BALB/c) recipient mice received titrated doses (5\u201020 x 106) of unseparated parental BALB/c BM without pharmacological NK cell activation. BM successfully engrafted in all mice and multilineage donor chimerism persisted long\u2010term (24\u00a0weeks), even in the absence of irradiation. Chimerism was associated with the rearrangement of the NK\u2010cell receptor repertoire suggestive of reduced reactivity to BALB/c. Chimerism levels were lower after transplantation with parental BALB/c than with syngeneic F1 BM, indicating partial NK\u2010mediated rejection of parental BM. Activation of NK cells with polyinosinic\u2013polycytidylic acid sodium salt poly(I:C), reduced parental chimerism in nonirradiated BM recipients but did not prevent hematopoietic stem cell engraftment. In contrast, equal numbers of parental lymph node cells were completely rejected. Hence, hybrid resistance leads to incomplete rejection of parental BM under reduced conditioning settings.Resistance to parental bone marrow ( Modifying conditioning in the hybrid resistance model reveals that lasting chimerism ensues after nonmyeloablative irradiation, even when no recipient irradiation is given, indicating that natural killer cell\u2013mediated bone marrow rejection is incomplete in reduced conditioning settings. BMbone marrowGvHDgraft versus host diseaseLNlymph nodeNKnatural killerpoly(I:C)polyinosinic\u2013polycytidylic acid sodium saltTBItotal body irradiationTLRtoll\u2010like receptor1Natural killer (NK) cells are large granular lymphocytes that serve as first\u2010line defense against pathogens and neoplastic cells.22.1CB6F1 and BALB/c (CD45.2) mice were purchased from Charles River and congenic B6.SJL\u2010Ptprca Pepcb/BoyJ (CD45.1) mice from Jackson Laboratory. F1 (CD45.1/CD45.2) mice were obtained by crossing male CD45.1 on C57BL/6 background with female CD45.2 BALB/c mice. All mice were housed under specific pathogen\u2010free conditions and female mice were used between 8 and 12\u00a0weeks of age. All animal experiments were approved by the internal review board of the Medical University of Vienna and by the Austrian Ministry of Science and Research (permission number GZ: BMWFW\u201066.009/0028\u2010WF/V/3b/2015).2.26) of unseparated BALB/c (CD45.2) or CB6F1 (CD45.2) BM cells (d0). Bones were flushed with a syringe and BM cells were collected in M199 medium (Sigma Aldrich) supplemented with 10\u00a0mM Hepes Buffer and 50\u00a0\u03bcg/ml gentamycin .F1 (CD45.1/CD45.2) recipient mice received titrated doses (5\u201020\u00a0\u00d7\u00a0102.36 BALB/c BM (d0) and poly(I:C) . Sixteen\u00a0weeks after transplantation, BM cells were recovered from primary recipients and transplanted into secondary F1 mice conditioned with 11\u00a0Gy TBI (2\u00a0\u00d7\u00a05.5\u00a0Gy). On the day of reconstitution, each secondary recipient was transplanted with 20\u00a0\u00d7\u00a0106 BM cells recovered from one chimera (i.v.).Primary recipients received 10\u00a0\u00d7\u00a0102.4Full\u2010thickness tail skin was grafted 4\u20106\u00a0weeks after BM transplantation and visually inspected thereafter at short intervals. Grafts were considered to be rejected when less than 10% remained viable, as described earlier.2.5\u2212 CD45.2+ cells among CD45.1+ CD45.2+ plus CD45.1\u2212 CD45.2+ leukocytes (CD45.1\u2212 CD45.2+/(CD45.1\u2212 CD45.2+ + CD45.1+ CD45.2+) \u00d7 100). APC anti\u2010mouse CD45.1 (A20), PE anti\u2010mouse CD45.2 (104), FITC anti\u2010mouse Mac\u20101 (M1/70), PE\u2010Cy7 anti\u2010mouse CD8 (53\u20106.7), APC\u2010Cy7 anti\u2010mouse CD4 (RM4\u20105), Pacific Blue anti\u2010mouse CD3 (17A2), FITC anti\u2010mouse CD49b (DX5), FITC anti\u2010mouse NK1.1 (PK136), PE anti\u2010mouse Ly49D (4E5), biotin anti\u2010mouse Ly49A (YE1/48.10.6) were purchased from Bio Legend. PE\u2010Cy7 anti\u2010mouse Ly49G2 (4D11) was purchased from eBioscience.The presence of donor cells was assessed at regular intervals by staining CD45.1 and CD45.2 on blood leukocytes. Donor chimerism was assessed as percentage of CD45.12.6t test with equal variances was used to compare chimerism levels. Total chimerism levels were compared between groups by using analysis of variance (ANOVA). The correlation between BM dose and chimerism level was assessed by a linear regression model. A P\u2010value below .05 was considered to denote statistical significance .Data were statistically analyzed with GraphPad Prism 5.0 . A 2\u2010sided Student's 36 BALB/c BM (d0). All recipients developed high levels of persistent multi\u2010lineage mixed chimerism that bind the very same MHC molecule.+ Ly49A/G2\u2212 NK cells male mice with BALB/c (CD45.2) female mice so that the resulting F1 generation coexpressed both CD45.1 and CD45.2, whereas donor BALB/c cells solely expressed CD45.2 7\u00a0days postinfusion by F1 recipient is given.Irradiation promotes engraftment by creating space in the BM niche,So far the proliferation of recipient splenocytes shortly after BM transplantation served as surrogate marker for BM engraftment in hybrid resistance models using lethally irradiated mice.+ NK cells would obtain the expression of the inhibitory receptors Ly49A and/or Ly49G2. This adaptation extended over a period of 4\u00a0weeks, which approximately corresponds to the time of NK cell maturation in the BM.Even if allogeneic stem cells have sufficient space to engraft, one would expect NK cells to resist their engraftment unless very large BM doses are infused.Our data from the murine hybrid resistance setting suggest that NK\u2010mediated BM rejection is less potent in reduced conditioning settings than in lethal irradiation regimens, allowing stem cell engraftment with moderate BM doses even in nonirradiated recipients.American Journal of Transplantation.The authors of this manuscript have no conflicts of interest to disclose as described by the"} +{"text": "A qu\u00adanti\u00adfication of the inter\u00admolecular contacts in the crystal were estimated using Hirshfeld surface analysis and two-dimensional fingerprint plots.2-(4-Nitro\u00adphen\u00adyl)-2-oxoethyl picolinate was synthesized under mild conditions. The chemical and mol\u00adecular structure was confirmed by single-crystal X-ray diffraction studies. The mol\u00adecules are related by inversion into centrosymmetric dimers 14H10N2O5, was synthesized under mild conditions. The chemical and mol\u00adecular structures were confirmed by single-crystal X-ray diffraction analysis. The mol\u00adecules are linked by inversion into centrosymmetric dimers via weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions, forming R22(10) ring motifs, and further strengthened by weak \u03c0\u2013\u03c0 inter\u00adactions. Hirshfeld surface analyses, the dnorm surfaces, electrostatic potential and two-dimensional fingerprint (FP) plots were used to verify the contributions of the different inter\u00admolecular inter\u00adactions within the supra\u00admolecular structure. The shape-index surface shows that two sides of the mol\u00adecules are involved with the same contacts in neighbouring mol\u00adecules and curvedness plots show flat surface patches that are characteristic of planar stacking.2-(4-Nitro\u00adphen\u00adyl)-2-oxoethyl picolinate, C The torsion angle value of \u03c42 = \u2212147.02\u2005(11)\u00b0 between the two carbonyl groups indicates a \u2013anti\u00adclinal conformation. Likewise, owing to a substitution on the functional group, the title com\u00adpound experiences steric repulsion between the substituent and adjacent carbonyl groups, which can influence the torsion angle [\u03c43 = 2.4\u2005(2)%] and resulting in a +synclinal conformation. The bond lengths and angles are normal and the mol\u00adecular conformation is characterized by a dihedral angle of 31.58\u2005(8)\u00b0 between the mean planes of the two aromatic rings. The nitro group lies nearly in the plane of the phenyl ring, as indicated by the torsion angle values of \u22124.7\u2005(2) and \u22125.1\u2005(2)\u00b0 for C4\u2014C3\u2014N1\u2014O4 and C2\u2014C3\u2014N1\u2014O5, respectively.The mol\u00adecular structure of the title com\u00adpound is shown in Fig.\u00a01B\u22efO3 inter\u00adactions stabilize the supra\u00admolecular architecture by forming Cg, N\u2014O\u22efCg and Cg\u22efCg inter\u00adactions. The hydrogen-bond geometry and lone pair-\u03c0 inter\u00adactions are listed in Table\u00a02Cg\u22efCg inter\u00adactions, i.e.Cg1\u22efCg1 and Cg2\u22efCg2 . These weak inter\u00admolecular inter\u00adactions link the mol\u00adecules to form a one-dimensional chain along the c axis and the mol\u00adecules exhibit layered stacking and shape-index (\u22121.0 to 1.0 a.u.), respectively. The calculated volume inside the Hirshfeld surface is 311.97\u2005\u00c53 in the area of 305.78\u2005\u00c53.Hirshfeld surfaces and fingerprint plots = 1.2 or 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019014105/jj2216sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019014105/jj2216Isup2.hklStructure factors: contains datablock(s) I. DOI: 1449658, 1449658CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Quantum-mechanical (QM) calculations were performed at the MP2/aug-cc-pVDZ//B3LYP/6-311++G level in vacuum phase, along with Bader\u2019s quantum theory of Atoms in Molecules (QTAIM). These processes involve transition states (TSs) with quasi-orthogonal structures (symmetry C1), which are highly polar, tight ion pairs \u2219. Gibbs free energies of activation for the A\u2219T(WC) / A\u2219T(rWC) \u2194 A*\u2219\u0422(rwWC) / A*\u2219\u0422(wWC) tautomeric transitions are lower than for the A\u2219T(H) / A\u2219T(rH) \u2194 A*N7\u2219\u0422(rwH) / A*N7\u2219\u0422(wH) tautomerisations . The (T)N3+H\u22efN1-(A), (T)O4+H\u22efN1-(A) / (T)N3+H\u22efN1-(A) and (T)O2+H\u22efN1-(A) H-bonds are found in the transition states TSA-\u00b7T+A\u00b7T(WC)\u2194A*\u00b7T(rwWC) / TSA-\u00b7T+A\u00b7T(rWC)\u2194A*\u00b7T(wWC). However, in the transition state TSA-\u00b7T+A\u00b7\u0422(H)\u2194A*N7\u00b7T(rwH) / TSA-\u00b7T+A\u00b7\u0422(rH)\u2194A*N7\u00b7T(wH), the (T)N3+H\u22efN7-(A), (T)O4+H\u22efN7-(A) / (T)N3+H\u22efN7-(A) and (T)O2+H\u22efN7-(A) H-bonds are supplemented by the attractive (T)O4+/O2+\u22efN6-(A) van der Waals contacts. It was demonstrated that the products of the tautomerization of the classical A\u2219T DNA base pairs\u2014A*\u2219\u0422(rwWC), A*N7\u2219\u0422(rwH) and A*N7\u2219\u0422(wH) (symmetry Cs)\u2013further transform via double proton transfer into the energetically favorable wobble A\u2219T*(rwWC), A\u2219T*(rwH) and A\u2219T*O2(wH) base mispairs (symmetry Cs).In this paper we have theoretically predicted a novel pathway for the mutagenic tautomerization of the classical A\u2219T DNA base pairs in the free state, the Watson-Crick A\u00b7\u0422(WC), reverse Watson-Crick A\u00b7\u0422(rWC), Hoogsteen A\u00b7\u0422(H) and reverse Hoogsteen A\u00b7\u0422(rH) pairs, Investigation of microstructural mechanisms for mutagenic tautomerization of the Watson-Crick DNA base pairs occupies an important place in molecular biophysics and molecular biology, enabling an understanding of the nature of genome instability \u20135. This N7 \u2013to N7 nitrogen atom; T*\u2013to O4 oxygen atom and T*O2 \u2013to O2 oxygen atom. L\u00f6wdin proposed that the A\u2219T(WC)\u2194A*\u2219T*(L) and G\u2219C(WC)\u2194G*\u2219C*(L) transitions occur by double proton transfer (DPT) along neighboring intermolecular hydrogen (H) bonds via proton tunneling. These ideas have been prominent in the field of quantum biology and attracted much theoretical study of the mechanisms of spontaneous transitions and transversions arising during DNA replication [L\u00f6wdin , 4 firstlication .kT for the G\u2219C(WC) DNA base pair [Recently, it has become clear that L\u00f6wdin\u2019s mechanism does not provide the generation of sufficiently long-lived mutagenic tautomers of the DNA bases, which escape from the replicative DNA-polymerase transforming into their canonical tautomeric forms. The root cause of this observation is the absence of the reverse barrier of tautomerization \u0394\u0394G in the A\u2219T(WC) DNA base pair and its small value in comparison with ase pair , 15\u201318.vice versa, which mechanism obviates the above difficulties. The chief difference of our mechanism from the L\u00f6wdin mechanism is that, in the process of mutagenic tautomerization through sequential proton transfer, the DNA bases shift laterally relative each other into the DNA minor or major grooves, leading to the wobble configuration which contains the mutagenic tautomers [In previous papers \u201327 we prautomers . Moreoveautomers , pyrimidautomers , 23 and automers \u201327 DNA bvice versa, which is the key to understanding the microstructural mechanisms for spontaneous transitions and transversions during DNA biosynthesis [This allows us to assume that it is the intrapair tautomeric transition of the wobble pairs from the main tautomeric form into the rare one with a WC configuration or close to it, and ynthesis \u201327. Theoynthesis \u201331.1) which are highly polar tight ion pairs \u2219.This paper uses QM/QTAIM methods to explore new pathways for mutagenic tautomerization of the classical Watson-Crick A\u00b7\u0422(WC), reverse Watson-Crick A\u00b7\u0422(rWC), Hoogsteen A\u00b7\u0422(H) and reverse Hoogsteen A\u00b7\u0422(rH) base pairs with a remarkable biological meaning . TheThe geometries of all the investigated DNA base pairs and transition states (TSs) were optimized using the Gaussian\u201909 package . The B3LWe have performed investigations for the isolated H-bonded pairs of nucleotide bases, that adequately reflects the processes occurring in real duplex environment , 30, 31.el = electronic energy, while Ecorr = thermal correction to Gibbs free energy.The Gibbs free energy G for all structures was obtained in the following way:int were calculated at the MP2/6-311++G level of theory as the difference between the total energy of the base pair and energies of the monomers and corrected for the basis set superposition error (BSSE) [Electronic interaction energies \u0394Er (BSSE) ,87 throur (BSSE) ,89 withor (BSSE) .Bader\u2019s quantum theory of Atoms in Molecules (QTAIM) \u201396 was aThe energies of the attractive van der Waals contacts , 102 in Energies of conventional AH\u00b7\u00b7\u00b7B H-bonds were evaluated by the empirical Iogansen formula :EAH\u00b7\u00b7\u00b7Bsonances \u2013114.The atom numbering scheme for the DNA bases is as per convention .These novel pathways for the mutagenic tautomerization of four biologically important A\u2219T DNA base pairs\u2014Watson-Crick A\u00b7\u0422(WC), reverse Watson-Crick A\u00b7\u0422(rWC), Hoogsteen A\u00b7\u0422(H) and reverse Hoogsteen A\u00b7\u0422(rH) \u201349\u2013are pvia proton transfer along intermolecular H-bonds as per currently known mechanisms for mutagenic tautomerization of WC pairs [Conformers of the A\u2219T base pairs remain plane symmetric structures along the entire IRC of tautomerization. This also holds for base pairs tautomerising WC pairs , 19, 49.WC) / A*\u2219\u0422(wWC) / A*N7\u2219\u0422(rwH) / A*N7\u2219\u0422(wH) tautomerisation reactions occur via the initial migration of proton localized at the N6 atom of the N6H2 amino group, leading to the formation of the A+\u2219\u0422- ion pair and significant change of the mutual orientation of the bases within the pair, i.e. mutual transformation of the cys / trans\u2194trans / cys-orientation of the N1H and N9H bonds relative to each other (1). Further proton transfers to the N1/N7 nitrogen atom causing the rotation of the base and formation of the terminal wobble base mispair. Each of these tautomeric conversions is followed by the asynchronous DPT along the intermolecular H-bonds in the wobble base mispairs (The A\u2219T(WC) / A\u2219T(rWC) / A\u2219T(H) / A\u2219T(rH) \u2194 A*\u2219\u0422(rwch other . Our newmispairs .WC) / A*\u2219\u0422(wWC) / A*N7\u2219\u0422(rwH) / A*N7\u2219\u0422(wH) tautomerisation reactions, the TSs are highly polar (~ 6.8\u201312.7 D) tight ion pairs \u2219 ion pairs. In the TSA-\u00b7T+A\u00b7T(WC)\u2194A*\u00b7T(rwWC) / TSA-\u00b7T+A\u00b7T(rWC)\u2194A*\u00b7T(wWC) transition states of tautomerisation the (T)N3+H\u22efN1-(A) (13.06 / 13.24) and (T)O4+ / O2+H\u22efN1-(A) are observed, while for the TSA-\u00b7T+A\u00b7\u0422(H)\u2194A*N7\u00b7T(rwH) / TSA-\u00b7T+A\u00b7\u0422(rH)\u2194A*N7\u00b7T(wH) transition states, the (T)N3+H\u22efN7-(A) (8.98 / 8.46) and (T)O4+ / O2+H\u22efN7-(A) (5.18 / 4.38) H-bonds are supplemented by attractive (T)O4+/O2+\u22efN6-(A) van der Waals contacts (+H\u22efN1-/N7-(A) H-bonds are significantly stronger than other specific contacts with increased ellipticity. The weakest among them are the attractive (T)O4+/O2+\u22efN6-(A) van der Waals contacts / A\u2219T(rWC) / A\u2219T(H) / A\u2219T(rH) \u2194 A*\u2219\u0422(rwl\u2219mol-1) . These Tcontacts , Table 2A*\u00b7T(rwWC)\u2194A\u00b7T*(rwWC), TSA*\u00b7T(wWC)\u2194A\u00b7T*O2(wWC), TSA*N7\u00b7\u0422(rwH)\u2194A\u00b7\u0422*(rwH) and TSA*N7\u00b7\u0422(wH)\u2194A\u00b7\u0422*O2(wH) of the DPT reactions are stabilized by the N6-H-N3 covalent bridge and one-single intermolecular H-bond\u2014N1H\u22efO4 (11.61), N1H\u22efO2 (10.94), N7H\u22efO4 (13.76) and N7H\u22efO4 , accordingly , which are characterized by low energies ECH\u00b7\u00b7\u00b7O, estimated by the Espinose-Molins-Lecomte formula [C\u22efO and dH\u22efO) in comparison with the canonical H-bonds BCPs of the intermolecular H-bonds range from 0.013 a.u. up to the 0.107 a.u.; the values of the Laplacian of the electron density \u0394\u03c1 at the BCPs are positive for all intrapair H-bonds and lie within a wide range from 0.005 a.u. up to the 0.152 a.u., demonstrating that H-bonds are attractive closed-shell ineractions; the value of the ellipticity \u03b5 varies in the range 0.79\u20138.6\u00b710\u22123 .A\u00b7\u00b7\u00b7B (2.574\u20133.103 \u00c5), dH\u00b7\u00b7\u00b7B (1.442\u20132.293 \u00c5) and \u2220AH\u22efB (139.2\u2013179.3\u00b0) .Interestingly, the energy of the intermolecular specific contacts constitute only a minor part of the electronic energy of monomeric interactions for all these H-bonded structures ~14\u20130.87%) /A*\u2219\u0422(wWC) tautomerisations is noticeably lower than for the A\u2219T(H)/A\u2219T(rH)\u2194A\u2219T*(rwH)/A\u2219T*O2(wH) tautomerisations , achieving maximum values for each tautomeric transition at its TS . The Gib2.65 D, aWC) reaction, occurs by participation of the dynamically unstable intermediate A*\u2219\u0422*(L) \u2194A*\u2219\u0422. TheWC), A*N7\u2219\u0422(rwH) and A*N7\u2219\u0422(wH) mispairs (s) tautomerise further via the DPT mechanism along neighboring intermolecular H-bonds into the energetically-favorable plane-symmetric A\u2219T*(rwWC), A\u2219T*(rwH) and A\u2219T*O2(wH) DNA base mispairs, respectively N3H\u22efN6(A) H-bonds, which, in fact, is a rate-limiting stage. It is noteworthy that the A*N7\u00b7\u0422(rwH)\u2192A\u00b7\u0422*(rwH) and A*N7\u00b7\u0422(wH)\u2192A\u00b7\u0422*O2(wH) tautomerisations are barrier-less (WC)\u2194A\u2219T*(rwWC) (1.39) and A*\u00b7T(wWC)\u2194A\u00b7T*O2(wWC) (1.77) are significantly lower than for the novel tautomerisation reactions , but are comparable with the values for the other DPT reactions [syn\u2194A\u2219G*syn [-1 for A\u2219T\u2194A*\u2219T* [It should be noted that three out of four tautomerization processes of the A\u2219T base pairs do not complete with formation of the A*\u2219\u0422(rwmispairs . These pectively and 2. Tl\u2219mol-1) , while teactions : from 2.\u2194A\u2219G*syn to 10.29\u2219T\u2194A*\u2219T* DPT taut\u041e2 [N7\u00b7\u0422(rwH)\u2194A\u00b7\u0422*(rwH) and A*N7\u00b7\u0422(wH)\u2194A\u00b7\u0422*O2(wH) tautomeric equilibria are completely shifted to the right. For the two other cases, the following proportions are observed: A*\u00b7\u0422(rwWC) (6.9%) \u2194 A\u00b7\u0422*(rwWC) (93.1%) and A*\u00b7\u0422(wWC) (83.6%) \u2194 A\u00b7\u0422*O2(wWC) (16.4%).It is thus possible to say that the tautomerization processes described here terminate with the mutagenic tautomerization of both T and A DNA bases with further formation of the classical mutagenic tautomers \u0422*, \u0422*\u041e2 , 106 and\u041e2 , 114, re-1) and populations of the investigated base mispairs yield the order: A\u00b7\u0422(H) (0.00) < A\u00b7\u0422(rH) (0.22/0.63) < A\u00b7\u0422(WC) (1.05/0.21) < A\u00b7\u0422(rWC) (1.31/0.16) < A\u2219T*(rwWC) (8.83/3.77\u221910\u22127) < A\u00b7\u0422*(rwH) (8.96/1.92\u221910\u22127) < A*\u2219T(rwWC) (10.07/2.47\u221910\u22128) < A*\u00b7T(wWC) (10.65/1.13\u221910\u22128) < A\u00b7T*O2(wWC) (12.31/2.07\u221910\u22129) < A\u00b7\u0422*O2(wH) (12.74/6.13\u221910\u221210) < A*\u2219T*(L) (13.51/1.95\u221910\u221210) < A*N7\u00b7\u0422(rwH) (24.69/1.08\u221910\u221218) < A*N7\u00b7\u0422(wH) (25.70/2.24\u221910\u221219). Notably, populations of the wobble A\u2219T*(rwWC), A\u00b7\u0422*(rwH), A*\u2219T(rwWC), A*\u00b7T(wWC), A\u00b7T*O2(wWC), A\u00b7\u0422*O2(wH) (12.74/6.13\u221910\u221210) and A*\u2219T*(L) tautomerised states, fitting into the range of the frequencies of the spontaneous point mutations observed experimentally (10\u221211\u201310\u22129) [Gibbs free energies \u2013117, poiNotably, the methyl group of the T DNA base does not change its orientation during all these tautomerisation processes without exception. Moreover, the heterocycles of the DNA bases remain planar, despite their ability for out-of-plane bending \u2013120.+ base occurs only in the TSA-\u00b7T+A\u00b7T(WC)\u2194A*\u00b7T(rwWC), TSA-\u00b7T+A\u00b7T(rWC)\u2194A*\u00b7T(wWC), TSA-\u00b7T+A\u00b7\u0422(H)\u2194A*N7\u00b7T(rwH) and TSA-\u00b7T+A\u00b7\u0422(rH)\u2194A*N7\u00b7T(wH) transition states. The maximum value of the non-planar dihedral angle reaches 2.5\u00b0 (C2-N3-C4-C5), 3.1\u00b0 (N1-C2-N3-C4), 3.7\u00b0 (C2-N3-C4-C5) and 7.8\u00b0 (N1-C2-N3-C4), respectively. Another structural feature of the protonated T+ base in these TSs is the deviation of the \u041e4+H / \u041e2+H hydroxyl group from the plane of the pyrimidine ring .1) are highly polar tight ion pairs \u2219. The tautomerization products\u2014the A*\u2219\u0422(rwWC), A*N7\u2219\u0422(rwH) and A*N7\u2219\u0422(wH) pairs\u2014further transform via concerted asynchronous double proton transfer into the energetically favorable wobble A\u2219T*(rwWC), A\u2219T*(rwH) and A\u2219T*O2(wH) mispairs (symmetry Cs), respectively. Moreover, it was established in our recent papers, that wobble A*\u2219T(rwWC) base mispair can also be formed from the reverse A\u2219T(rWC) base pair [N7\u00b7\u0422(wH) base mispair\u2014from the Hoogsteen A\u2219T(H) base pair [Novel pathways for mutagenic tautomerization of four classical A\u2219T DNA base pairs, followed by the significant changes of base orientation within the pair, have been predicted by these QM results. The transition states with quasi-orthogonal structure Click here for additional data file."} +{"text": "CSNK2A1 inhibited TGF\u03b2\u2010induced EMT. TGF\u03b2 signaling decreased CK2\u03b2 but did not affect CK2\u03b1 protein levels, resulting in a quantitative imbalance between the catalytic \u03b1 and regulatory \u03b2 subunits, thereby increasing CK2 activity. The decrease in CK2\u03b2 expression was dependent on TGFBRI kinase activity and the ubiquitin\u2013proteasome pathway. The E3 ubiquitin ligases responsible for TGF\u03b2\u2010induced CK2\u03b2 degradation were found to be CHIP and WWP1. Okadaic acid (OA) pretreatment protected CK2\u03b2 from TGF\u03b2\u2010induced degradation, suggesting that dephosphorylation of CK2\u03b2 by an OA\u2010sensitive phosphatase might be required for CK2 activation in TGF\u03b2\u2010induced EMT. Collectively, our results suggest CK2 as a therapeutic target for the prevention of EMT and metastasis of cancers.Transforming growth factor \u03b2 (TGF\u03b2) is overexpressed in advanced cancers and promotes tumorigenesis by inducing epithelial\u2013mesenchymal transition (EMT), which enhances invasiveness and metastasis. Although we previously reported that EMT could be induced by increasing CK2 activity alone, it is not known whether CK2 also plays an essential role in TGF\u03b2\u2010induced EMT. Therefore, in the present study, we investigated whether TGF\u03b2 signaling could activate CK2 and, if so, whether such activation is required for TGF\u03b2\u2010induced EMT. We found that CK2 is activated by TGF\u03b2 treatment, and that activity peaks at 48\u00a0h after treatment. CK2 activation is dependent on TGF\u03b2 receptor (TGFBR) I kinase activity, but independent of SMAD4. Inhibition of CK2 activation through the use of either a CK2 inhibitor or shRNA against CHIPcarboxyl terminus of Hsc70\u2010interacting proteinCK2protein kinase CK2CKDCSNK2A1 knockdownEMTepithelial\u2013mesenchymal transitionHEKhuman embryonic kidneyNSCLCnon\u2010small cell lung cancerOAokadaic acidSKDSMAD4\u2010knockdownTGFBRIITGF\u03b2 type II receptorTGFBRITGF\u03b2 type I receptorTGF\u03b2transforming growth factor \u03b2WWP1WW domain containing E3 ubiquitin protein ligase 11et\u00a0al., et\u00a0al., et\u00a0al., et\u00a0al., et\u00a0al., et\u00a0al., et\u00a0al., et\u00a0al., et\u00a0al., et\u00a0al., et\u00a0al., et\u00a0al., Transforming growth factor \u03b2 (TGF\u03b2) is a potent pleiotropic cytokine that regulates cell growth/differentiation, cell motility, extracellular matrix production, angiogenesis and cellular immune responses , and 100\u00a0U\u00b7mL\u22121 penicillin (Gibco). Human embryonic kidney (HEK) 293 cells were cultured in Dulbecco's modified Eagle's medium (Gibco). All cells were cultured at 37\u00a0\u00b0C in 5% CO2. The CK2 inhibitor, emodin , and the reversible cell\u2010permeable proteasome inhibitor, MG132 (Sigma\u2010Aldrich), were prepared in 20\u00a0mm stock with dimethylsulfoxide (Sigma\u2010Aldrich). The TGFBRI\u2010specific inhibitor, SB431542 (Sigma\u2010Aldrich), was prepared in 10\u00a0mm stock with dimethylsulfoxide. The protein phosphatase inhibitor, okadaic acid , was prepared in 10\u00a0\u03bcm stock with dimethylsulfoxide. TGF\u03b2 was prepared in 10\u00a0\u03bcg\u00b7mL\u22121 stock. pCMV5 TBRI\u2010His was a gift from Joan Massague (Addgene plasmid # 19161). pCMV5B\u2010TGF\u03b2 receptor I K232R was a gift from Jeff Wrana (Addgene plasmid # 11763). p3TP\u2010Lux gene TGF\u03b2 response element and three collagenase I AP\u20101 repeats in front of luciferase, was a gift from Joan Massague & Jeff Wrana (Addgene plasmid # 11767). pCMV5B\u2010Flag\u2010Smurf1 wt was a gift from Jeff Wrana (Addgene plasmid # 11752), pCMV5B\u2010Flag\u2010Smurf2 wt was a gift from Jeff Wrana (Addgene plasmid # 11746), p4489 Flag\u2010betaTrCP was a gift from Peter Howley (Addgene plasmid # 10865), pcDNA3\u2010HA2\u2010ROC1 was a gift from Yue Xiong (Addgene plasmid # 19897) and pCI HA NEDD4 was a gift from Joan Massague (Addgene plasmid # 27002). pCMV\u2010Tag3B\u2010WWP1\u2010myc was kindly provided by Dr Ceshi Chen (Kunming Institute of Zoology). Flag\u2010tagged pCMV\u2010Tag2C\u2010WWP1 was kindly provided by Dr Hyeon Soo Kim cell line, A549 was cultured in Roswell Park Memorial Institute 1640 medium , supplemented with 10% fetal bovine serum (Gibco), 100\u00a0\u03bcg\u00b7mL2.2et\u00a0al., Western blot analysis was performed as described previously . The beads were incubated with 100\u00a0\u03bcg of cell lysates in a final volume of 50\u00a0\u03bcL of kinase reaction buffer of [\u03b3\u201032P]\u2010ATP]) for 20\u00a0min at 30\u00a0\u00b0C. The reactions were stopped by washing twice with 1\u00a0\u00d7\u00a0kinase buffer. The samples were resuspended with 30\u00a0\u03bcL of 2\u00a0\u00d7\u00a0SDS/PAGE sample\u2010loading buffer, subjected to 12% SDS/PAGE, stained with Coomassie Brilliant Blue, and dried on Whatman paper . 32P incorporation was detected by autoradiography.To evaluate intracellular CK2 activity, an 2.4CSNK2A1 . The shRNA sequences tested for SMAD4 knockdown were: sequence #1: TTCAGGTGGCTGGTCGGAAAGGATTTCCT; sequence #2: GCAGCCATAGTGAA GGACTGTTGCAGATA; sequence #3: CCAACATTCCTGTGGCTTCCACAAGTC AG; and sequence #4: GTCAGGTGCCTTAGTGACCACGCGGTCTT. We validated all constructs individually and found that constructs #1 and #2 were effective for SMAD4 knockdown. Subsequently, we used construct #1 for SMAD4 knockdown. Mission\u00ae esiRNA human STUB1 was purchased from Sigma\u2010Aldrich. Silencer\u00ae select pre\u2010designed siRNA targeting human WWP1 was purchased from Ambion (Thermo Fisher Scientific). The cells were transfected with siRNA using Lipofectamine\u00ae RNAiMAX in accordance with the manufacturer's instructions.shRNA\u2010mediated knockdown of 2.5The cells were seeded in six\u2010well plates and cotransfected with p3TP\u2010Lux and pRL\u2010TK using ViaFect\u2122 . Twenty\u2010four hours after transfection, the cells were treated with TGF\u03b2 for 24\u00a0h, washed with PBS and harvested. Cell lysates were prepared with 200\u00a0\u03bcL of Passive Lysis buffer (Promega). Aliquots (20\u00a0\u03bcL) of cleared lysate were analyzed for luciferase activity using a Dual\u2010luciferase\u00ae reporter assay system (Promega). The luciferase activity of p3TP\u2010Lux was normalized to that of pRL\u2010TK.2.6m Hepes (pH 7.9), 10\u00a0mm KCl, 0.1\u00a0mm EDTA, 1\u00a0mm dithiothreiltol, 1\u00a0mm phenylmethanesulfonyl fluoride, 1\u00a0\u00d7\u00a0protease inhibitor cocktail and 1\u00a0mm sodium orthovanadate. The samples were adjusted to 0.6% Nonidet P\u201040 (NP\u201040), and vortexed vigorously for 10\u00a0s. Nuclei were pelleted by centrifugation at 10\u00a0000\u00a0\u00d7\u00a0g for 30\u00a0s at 4\u00a0\u00b0C. The supernatants were collected and used as the cytoplasmic fraction. After washing the pellets with PBS, they were lysed in buffer C comprising 20\u00a0mm Hepes, pH 7.9, 0.4\u00a0m NaCl, 0.1\u00a0mm EDTA, 1\u00a0mm dithiothreitol, 1\u00a0mm phenylmethanesulfonyl fluoride, 1\u00a0\u00d7\u00a0protease inhibitor cocktail and 1\u00a0mm sodium orthovanadate by sonication. The lysates were cleared by centrifugation at 10\u00a0000\u00a0\u00d7\u00a0g for 20\u00a0min at 4\u00a0\u00b0C. The supernatants were collected and used as the nuclear fraction.The cells were allowed to swell in buffer A comprising 10\u00a0m2.7m Tris\u2010HCl, pH 7.4, 150\u00a0mm NaCl, 0.5% NP\u201040) with cOmplete\u2122 protease inhibitor cocktail (Roche Diagnostics). The cell lysates were pre\u2010cleared and then incubated with the appropriate antibodies for 1\u00a0h at 4\u00a0\u00b0C. The antibody\u2013protein complexes were precipitated with Protein A/G\u2010Sepharose beads (Santa Cruz Biotechnology Inc.), washed, and resuspended in 40\u00a0\u03bcL of SDS/PAGE loading buffer.The cells were collected and lysed with 1\u00a0mL of immunoprecipitation lysis buffer mutated to non\u2010phosphorylatable alanine residues or to phosphomimetic glutamic acids or to generate TGFBRI constitutively active (CA) mutant (threonine 204 is replaced with aspartic acid), mutagenesis was performed using a QuikChange site\u2010directed mutagenesis kit . All mutant constructs were confirmed by DNA sequencing. The mutagenic primer pairs used to generate mutants were: CK2\u03b2 3E (S2ES3ES4E): forward, 5\u2032\u2010GACGTGAAGATGGAAGAAGAAGAGGAGGTGTCC\u20103\u2032; reverse, 5\u2032\u2010GGACACCTCCTCTTCTTCTTCGATCTTCACGTC\u20103\u2032. CK2\u03b2 3A (S2A S3AS4A): forward, 5\u2032\u2010GACGTGAAGATGGCAGCAGCAGAGGAGGTGTCC\u20103\u2032; reverse, 5\u2032\u2010GGACACCTCCTCTGCTGCTGCCATCTTCACGTC\u20103\u2032. TGFBRI CA: forward, 5\u2032\u2010GAACAATTGCGAGAGATATTGTGTTACAAG\u20103\u2032; reverse, 5\u2032\u2010TCCGTA ACACAATATCTCTCGCAATTGTTC\u20103\u2032.2.9A cell migration assay was conducted using specific wound\u2010assay chambers purchased from ibidi GmbH . All experiments were performed in accordance with the manufacturer's instructions.2.10t\u2010test. P\u00a0<\u00a00.05 was considered statistically significant.Statistical comparisons of groups were performed using Student's 33.1et\u00a0al., et\u00a0al., in\u00a0vitro kinase assay and western blot analysis were performed. CK2 activity peaked at 48\u00a0h after TGF\u03b2 treatment and the E\u2010to N\u2010cadherin switch was observed 24\u00a0h after TGF\u03b2 treatment A549 cells. Previously, we reported that CKD could decrease cellular CK2 activity A549 cells using shRNA. When SKD cells were treated with TGF\u03b2, EMT was induced and CK2 was activated A549 cells were generated using the CRISPR/Cas9 gene knockout system. Western blot analysis and in\u00a0vitro kinase assay showed that with \u03b2KO, the E\u2010 to N\u2010cadherin switch was induced . In the present study, we treated cells with 2\u00a0nm OA and thus PP2A could be completely inhibited; however, this might not be the case for PP1. OA treatment protected CK2\u03b2 from TGF\u03b2\u2010induced degradation . Further experiments, including the generation of PPP2CA and PPP2CB double knockout A549 cells, are required to identify the phosphatase involved in TGF\u03b2\u2010induced CK2\u03b2 dephosphorylation.In non\u2010canonical TGF\u03b2 signaling, TGFBRI kinase\u2010dependent activation and interaction of phosphatase 2A with p70\u2010S6 kinase could result in the dephosphorylation and inactivation of the kinase, thereby inducing G1 arrest on motility.Click here for additional data file.Fig.\u00a0S2. Polyubiquitination of CK2\u03b2 by TGF\u03b2 signaling.Click here for additional data file.Fig.\u00a0S3. Screening of E3 ubiquitin ligases for CK2\u03b2 degradation.Click here for additional data file."} +{"text": "A, B, C and D). The conformations of the mol\u00adecules differ, as seen from the dihedral angles between the two pyridine rings in each mol\u00adecule. They vary from 5.51\u2005(9)\u00b0 for mol\u00adecule B to 25.25\u2005(8)\u00b0 for mol\u00adecule A.The asymmetric unit of the title disubstituted 2,3\u2032-bi\u00adpyridine, contains four independent mol\u00adecules in the asymmetric unit. The dihedral angles between the two pyridine rings in each mol\u00adecule are 25.25\u2005(8)\u00b0 in A, 5.51\u2005(9)\u00b0 in B, 11.11\u2005(9)\u00b0 in C and 16.24\u2005(8)\u00b0 in D. In the crystal, mol\u00adecules A and B are linked by C\u2014H\u22efN hydrogen bonds to form layers extending parallel to the ab plane, while mol\u00adecules C and D are linked by C\u2014H\u22efN hydrogen bonds forming \u2013C\u2013D\u2013C\u2013D\u2013 chains propagating along the b-axis direction. The layers and the chains are stacked alternately along the c axis through offset \u03c0\u2013\u03c0 and C\u2261N\u22ef\u03c0 [N-to-pyridine-centroid distance = 3.882\u2005(2)\u2005\u00c5] inter\u00adactions, resulting in the formation of a supra\u00admolecular framework.The title compound, C T1) compared with phenyl\u00adpyridine-based CT1: 2.82\u2005eV) is larger than that of alk\u00adoxy-functionalized analogue, 2\u2032,6\u2032-dimeth\u00adoxy-2,3\u2032-bi\u00adpyridine (T1: 2.70 eV) research area because developing blue phospho\u00adrescent materials remains a problem that has not been solved so far. Although there are a number of advantages in 2,3\u2032-bi\u00adpyridine ligands, incorporating the substituents into the ligand framework is difficult owing to the low selectivity and reactivity of the pyridine ring . The dihedral angles between the two pyridine rings in each mol\u00adecule are 25.25\u2005(8)\u00b0 in A, 5.51\u2005(9)\u00b0 in B, 11.11\u2005(9)\u00b0 in C and 16.24\u2005(8)\u00b0 in D. In order to investigate the conformational similarity between the four mol\u00adecules, the r.m.s. overlay fits of the 16 non-H atoms of each mol\u00adecule were calculated using the AutoMolFit routine in PLATON , forming layers extending parallel to the ab plane, while the C and D mol\u00adecules are connected through C\u2014H\u22efN hydrogen bonds to from \u2013C\u2013D\u2013C\u2013D\u2013 chains propagating along the b-axis direction. The layers and chains stack alternately along the c axis, linked by inter\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions, resulting in the formation of a supra\u00admolecular framework, as shown in Fig.\u00a04Cg1\u22efCg2Di = 3.6741\u2005(9)\u2005\u00c5; Cg1\u22efCg2Div = 3.6546\u2005(9)\u2005\u00c5; Cg2\u22efCg1Div = 3.5888\u2005(9)\u2005\u00c5; Cg2B\u22efCg1Civ = 3.8196\u2005(10)\u2005\u00c5; Cg1 and Cg2 are the centroids of the N1/C1\u2013C5 and N2/C6-C10 rings. Atoms and centroids labelled with suffixes B, C and D represent those of the mol\u00adecules B, C and D, respectively]. In addition, inter\u00admolecular C\u2261N\u22ef\u03c0 inter\u00adactions between the cyano N atom of the D mol\u00adecule and the N1B-containing pyridine ring of mol\u00adecule B + .All experiments were performed under a dry NUiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018011532/xu5937sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989018011532/xu5937Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018011532/xu5937Isup3.cmlSupporting information file. DOI: 1862117CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A and B) in the asymmetric unit. They differ essentially in the conformation of the pyrrolidine and cyclo\u00adpentene rings; respectively, twisted and flat in mol\u00adecule A, but envelope and twisted in mol\u00adecule B.The title compound crystallized with two independent mol\u00adecules in the asymmetric unit. In both mol\u00adecules, the pyran and pyridine rings adopt envelope and chair conformations, respectively. The conformation of the pyrrolidine and cyclo\u00adpentene rings differ in the two mol\u00adecules; twisted and flat, respectively, in mol\u00adecule A, but envelope and twisted, respectively, in mol\u00adecule B. In both mol\u00adecules, there is a C\u2014H\u22efN intra\u00admolecular hydrogen bond present. In both mol\u00adecules, the oxygen atoms of the nitro groups are disordered as is the chlorine atom in mol\u00adecule B. In the crystal, the B mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, forming \u2013B\u2013B\u2013B\u2013 chains along [010], and by C\u2014H\u22ef\u03c0 inter\u00adactions. The A and B mol\u00adecules are also linked by a number of C\u2014H\u22ef\u03c0 inter\u00adactions, resulting in the formation a supra\u00admolecular three-dimensional structure. In mol\u00adecule A, the nitro group oxygen atoms are disordered over two positions with refined occupancy ratios of the nitro group oxygen atoms O3A and O4A in 0.59\u2005(2):0.41\u2005(2) while in molecule B one of the nitro O atoms is disordered over two positions with a refined occupancy ratio of 0.686\u2005(13):0.314\u2005(13) and the chlorine atoms is disordered over two positions with a refined occupancy ratio of 0.72\u2005(3):0.28\u2005(3).The title compound, C The mean plane of the five-membered pyrrolidine ring (N1/C12/C13/C21/C22) is inclined to the mean plane of the cyclo\u00adpentene ring (C1/C2/C10\u2013C12) by 87.30\u2005(13) and 88.41\u2005(11)\u00b0 in mol\u00adecules A and B, respectively. The benzene rings C27\u2013C32 and C14\u2013C19 are inclined to each other by 58.13\u2005(13)\u00b0 in mol\u00adecule A and 57.13\u2005(11)\u00b0 in mol\u00adecule B, while benzene rings C6\u2013C11 and C3\u2013C5/C33\u2013C35 are inclined to each other by 10.20\u2005(13)\u00b0 in mol\u00adecule A and 4.08\u2005(13)\u00b0 in mol\u00adecule B. The mean plane of the pyrrolidine ring (N1/C12/C13/C21/C22) makes a dihedral angle with the mean plane of the pyran ring (O2/C13/C14/C19\u2013C21) of 34.6\u2005(2)\u00b0 in mol\u00adecule A and 29.65\u2005(10)\u00b0 in mol\u00adecule B, and is inclined to the piperidine ring mean plane (N1/C22\u2013C26) by 15.69\u2005(12)\u00b0 in mol\u00adecule A and 12.36\u2005(11)\u00b0 in mol\u00adecule B. The mean planes of the pyran and piperidine rings are inclined to each other by 37.06\u2005(11) and 29.49\u2005(10)\u00b0 in mol\u00adecules A and B, respectively. The mean plane of the pyrazine ring (N3/N4/C1/C2/C3/C4) makes a dihedral angle with the mean plane of the pyran ring (O2/C13/C14/C19\u2013C21) of 63.42\u2005(19)\u00b0 in mol\u00adecule A and 72.64\u2005(10)\u00b0 in mol\u00adecule B. It is inclined to the pyrrolidine ring mean plane (N1/C12/C13/C21/C22) by 88.11\u2005(1)\u00b0 in mol\u00adecule A and 86.69\u2005(11)\u00b0 in mol\u00adecule B and is inclined to the piperidine ring mean plane (N1/C22\u2013C26) by 77.24\u2005(11)\u00b0 in mol\u00adecule A and 82.97\u2005(11)\u00b0 in mol\u00adecule B.Chlorine atoms Cl1B mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds, forming chains propagating along the b-axis direction -6a-nitro-6,6a,6\u2005b,7,8,9,10,12a-octa\u00adhydro\u00adspiro\u00adindolizine-12,3-indolin]-2-one under a nitro\u00adgen atmosphere. The solution was refluxed for 20\u2005h in a Dean\u2013Stark apparatus to give the corresponding cyclo\u00adadduct. After completion of the reaction, as indicated by TLC, the solvent was evaporated under reduced pressure. The crude product obtained was purified by column chromatography using hexa\u00adne/EtOAc (6:4) as eluent (yield 86%). Colourless block-like crystals of the title compound were obtained by slow evaporation of a solution in ethanol.To a solution of indeno\u00adquinoxalinone (1.0\u2005mmol) and pipacolinic acid (1.5\u2005mmol) in dry toluene, was added 2-(4-chloro\u00adphen\u00adyl)-3-nitro-2Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a02A and O4A in A and O3B and O4B in B are disordered over two positions with refined occupancy ratios of O3A/O4A:O3A\u2032/O4A\u2032 = 0.59\u2005(2):0.41\u2005(2), and O3B/O4B:O3B\u2032/O4B\u2032 = 0.686\u2005(13):0.314\u2005(13). In mol\u00adecule B, the chlorine atom Cl2 is disordered over two positions with a refined occupancy ratio of Cl2:Cl2\u2032 = 0.72\u2005(3):0.28\u2005(3).In both mol\u00adecules, the nitro group oxygen atoms O310.1107/S2056989019000975/su5475sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019000975/su5475Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019000975/su5475Isup3.cmlSupporting information file. DOI: 1024832CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Out-of-hospital electrocardiogram (ECG) shows a\u00a0sinus rhythm of 85\u00a0beats per minute (bpm), narrow QRS complex, and ST-segment elevations in leads V1\u2013V4 (coved morphology in V1\u2013V2), with a\u00a0terminal negative T\u00a0wave in V1\u2013V2 (Fig.\u00a0The patient met criteria for exertional heat stroke , and wasDiffuse ST-T deviations have been described in patients with heat stroke , but rigType\u00a01 Brugada-like ECG pattern induced by exertional heat stroke."} +{"text": "The benzo\u00adthia\u00adzine skeleton is not planar, with a maximum deviation of 0.3154\u2005(11)\u2005\u00c5 from the least-squares plane. The mol\u00adecule was expected to adopt mirror symmetry but slightly different conformational characteristics of the condensed benzo\u00adthia\u00adzine ring lead to point group symmetry 1. 19H14N2O6S2\u00b7C3H7NO. The mol\u00adecule was expected to adopt mirror symmetry but slightly different conformational characteristics of the condensed benzo\u00adthia\u00adzine ring lead to point group symmetry 1. In the crystal, mol\u00adecules form two types of stacking dimers with distances of 3.464\u2005(2)\u2005\u00c5 and 3.528\u2005(2)\u2005\u00c5 between \u03c0-systems. As a result, columns extending parallel to [100] are formed, which are connected to inter\u00admediate di\u00admethyl\u00adformamide solvent mol\u00adecules by C\u2014H\u22efO inter\u00adactions.The title mol\u00adecule crystallizes as a di\u00admethyl\u00adformamide monosolvate, C R-4-hy\u00addroxy-2-oxo-1,2-di\u00adhydro\u00adquinoline-3-carboxyl\u00adates are highly reactive compounds -one 6,6,8,8-tetroxide (I)H-2\u03bb6,1-benzo\u00adthia\u00adzine-3-carb\u00adoxyl\u00adate and S = 0.48, \u03b8 = 50.0\u00b0, \u03a8 = 22.8\u00b0 for the C5\u2013C6\u2013C7\u2013C8\u2013N2\u2013S2 ring (2). The S1 and C1 atoms deviate by 0.669\u2005(2) and 0.207\u2005(2)\u2005\u00c5, respectively, from the mean-square plane of the remaining atoms in ring (1). The corresponding deviations in ring (2) are 0.668\u2005(2) and 0.270\u2005(2)\u2005\u00c5, respectively.Both thia\u00adzine rings adopt a twist-boat conformation Fig.\u00a01 with sliH-pyran-4-one ring (3) adopts a sofa conformation with puckering parameters S = 0.14, \u03b8 = 24.7\u00b0, \u03a8 = 22.6\u00b0. The deviation of C19 from the plane of the remaining atoms of (3) is 0.087\u2005(2)\u2005\u00c5. The C1=C2 and C5=C6 bonds [1.3571\u2005(17)\u2005\u00c5 and 1.3529\u2005(17)\u2005\u00c5] are slightly elongated as compared to the mean value of 1.329\u2005\u00c5 for a Csp2=Csp2 bond , H13\u22efH18B = 2.28\u2005\u00c5 (expected 2.34\u2005\u00c5), H13\u22efH18C = 2.31\u2005\u00c5 (expected 2.34\u2005\u00c5). These shortened contacts affect the very small pyramidalization of the nitro\u00adgen atoms; the sums of the bond angles centered at the N1 and N2 atoms are 354 and 356\u00b0, respectively.A further analysis of the mol\u00adecular structure revealed the presence of other shortened intra\u00admolecular contacts: H9\u22efH17In the crystal, mol\u00adecules of (I)Ai and C18\u2014H18B\u22efO1Aii and diphenyl oxide (10\u2005ml) was maintained on a metal bath at 493\u2005K for 3\u2005h, then cooled and diluted with ethanol = 1.5Ueq(C) for methyl groups and with C\u2014H = 0.93\u2005\u00c5, Uiso(H) = 1.2Ueq(C) for all other hydrogen atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019008788/wm5508sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019008788/wm5508Isup2.hklStructure factors: contains datablock(s) I. DOI: 1935426CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Conventional O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonding leads to a supra\u00admolecular layer in the crystal.A coordination geometry inter\u00admediate between square-planar and tetra\u00adhedral, defined by an N II complex, isolated as a dihydrate, [Cu(C21H14N3O3)2]\u00b72H2O, reveals a highly distorted coordination geometry inter\u00admediate between square-planar and tetra\u00adhedral defined by an N2O2 donor set derived from two mono-anionic bidentate ligands. Furthermore, each six-membered chelate ring adopts an envelope conformation with the Cu atom being the flap. In the crystal, imidazolyl-amine-N\u2014H\u22efO(water), water-O\u2014H\u22efO, phenyl-C\u2014H\u22efO(nitro) and \u03c0(imidazol\u00adyl)\u2013\u03c0(nitro\u00adbenzene) [inter-centroid distances = 3.7452\u2005(14) and 3.6647\u2005(13)\u2005\u00c5] contacts link the components into a supra\u00admolecular layer lying parallel to (101). The connections between layers forming a three-dimensional architecture are of the types nitro\u00adbenzene-C\u2014H\u22efO(nitro) and phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl). The distorted coordination geometry for the CuII atom is highlighted in an analysis of the Hirshfeld surface calculated for the metal centre alone. The significance of the inter\u00admolecular contacts is also revealed in a study of the calculated Hirshfeld surfaces; the dominant contacts in the crystal are H\u22efH (41.0%), O\u22efH/H\u22efO (27.1%) and C\u22efH/H\u22efC (19.6%).The crystal and mol\u00adecular structures of the title Cu Complex (I)via an analysis of the calculated Hirshfeld surfaces.The title copper(II) complex, (I)N,O-chelated by two 2--4-nitrophenolate mono-anions. The resulting N2O2 donor set defines a highly distorted coordination geometry, as seen in the angles included in Table\u00a014 is a geometric measure of the distortion of a four-coordinate geometry \u2005\u00c5 out of the plane. The comparable parameters for the O2-chelate ring are 0.033 and 0.354\u2005(3)\u2005\u00c5, respectively. The dihedral angle formed between the two planar regions of the chelate rings is 49.38\u2005(8)\u00b0. The dihedral angles between the best plane through the O1-chelate ring and each of the fused six- and five-membered rings are 9.18\u2005(12) and 5.54\u2005(14)\u00b0, respectively; the equivalent angles for the O2-chelate rings are 8.44\u2005(8) and 2.71\u2005(9)\u00b0, respectively. The N1-imidazol-2-yl ring forms dihedral angles of 41.20\u2005(11) and 37.46\u2005(10)\u00b0 with the C10- and C16-phenyl substituents, respectively, and the dihedral angle between the phenyl rings is 59.92\u2005(8)\u00b0, i.e. all indicating splayed relationships. A similar situation pertains to the N2-imidazol-2-yl ring, where the comparable dihedral angles formed with the C31- and C37-phenyl rings are 38.29\u2005(10), 48.5\u2005(9) and 50.84\u2005(7)\u00b0, respectively. Finally, the nitro groups are not strictly coplanar with the benzene rings to which they are connected, as seen in the dihedral angles of 14.2\u2005(4)\u00b0 for C1\u2013C6/N4/O3/O4 and 5.9\u2005(3)\u00b0 for C22\u2013C27/N6/O5/O6.The crystallographic asymmetric unit of (I)W water mol\u00adecule forms donor inter\u00adactions to the coordinated O2 atom and to a symmetry-related O2W water mol\u00adecule. The O2W water mol\u00adecule connects to the coordinated O1 atom as well as to a nitro-O3 atom. Hence, the O2W water mol\u00adecule is involved in four hydrogen-bonding inter\u00adactions. The fourth contact involving the O1W water mol\u00adecule, a C\u2014H\u22efO acceptor contact, is provided by the nitro\u00adbenzene ring. There is also a phenyl-C\u2014H\u22efO(nitro) contact of note, Table\u00a02a). There are also \u03c0\u2013\u03c0 stacking and C\u2014H\u22efO inter\u00adactions in the crystal, Fig.\u00a03b). Within layers, there are \u03c0\u2013\u03c0 inter\u00adactions occurring between the imidazolyl and nitro\u00adbenzene rings . The connections between layers along [010] are of the type nitro\u00adbenzene-C\u2014H\u22efO(nitro) and phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl), as detailed in Table\u00a02As each component of the asymmetric unit has hydrogen-bonding functionality, conventional hydrogen bonds are found in the crystal of (I)CrystalExplorer17 , and O2W-water, Fig.\u00a04b), mol\u00adecules. In addition, the presence of faint-red spots near the O1W, O2W and H1W atoms in Figs. 4a) and 4(b) are indicative of the other contacts of these atoms with those of the CuII complex mol\u00adecule (Table\u00a02c)\u2013(e).The Hirshfeld surface calculations for (I)e Table\u00a02. The dondnorm-mapped Hirshfeld surface in Fig.\u00a04c). The pair of faint-red spots appearing near the phenyl-C36 and H36 atoms, and also near the nitro-O5 atom on the surface indicating short inter\u00adatomic contacts that characterize the weak C\u2014H\u22efO inter\u00adaction, Table\u00a03e). The donors and acceptors of this inter\u00adaction are also evident as the blue bump and a bright-orange spot enclosed within the black circle on the Hirshfeld surface mapped with the shape-index property in Fig.\u00a05a). The bright-orange region enclosed within a black circle in Fig.\u00a05b) is also an indication of the O2W\u2014H4W\u22efCg(C16\u2013C21) contact. The Hirshfeld surfaces mapped over the calculated electrostatic potential for the water and complex mol\u00adecules in Fig.\u00a06dnorm illustrated in Figs.\u00a04c)\u2013(e), faint-red spots also appear near other atoms indicating their involvement in other short inter\u00adatomic contacts, as summarized in Table\u00a03The presence of a short inter\u00adatomic C\u22efC contact between atoms C22 and C28 Table\u00a03 arises f2O4 donor set about the copper(II) centre in the complex mol\u00adecule. This is performed by considering the Hirshfeld surface about the metal centre alone and the small orange regions on the surface relatively far from the Cu\u2014N bonds in Fig.\u00a08b). The different curvature of the Hirshfeld surfaces coordinated by the N2O4 donor set in Figs.\u00a08c) and 8(d) also support this observation. The Cu\u2014O and Cu\u2014N bonds are rationalized in the two-dimensional fingerprint plot taking into account only the Hirshfeld surface for the copper atom shown in Fig.\u00a09de + di \u223c 2.0\u2005\u00c5 for the Cu\u2014N bonds (upper region) and the Cu\u2014O bonds (lower region) are indicative of the distorted geometry , and those delineated into H\u22efH, O\u22efH/H\u22efO, C\u22efH/H\u22efC, C\u22efC and C\u22efO/O\u22efC contacts are illustrated in Figs.\u00a010b)\u2013(f), respectively. The percentage contribution from different inter\u00adatomic contacts to the Hirshfeld surfaces of the complex mol\u00adecule and for overall (I)de + di \u223c 1.9\u2005\u00c5 in the fingerprint plot delineated into H\u22efH contacts shown in Fig.\u00a010b) is the result of the involvement of the H12 atom in a short inter\u00adatomic H\u22efH contact, Table\u00a03W and H2W, Table\u00a03de\u00a0+ di \u223c 1.8\u2005\u00c5 in the plot delineated into O\u22efH/H\u22efO contacts, Fig.\u00a010c), arise from the N\u2014H\u22efO hydrogen bond, while the merged points correspond to other inter\u00adactions at greater inter\u00adatomic distances. The significant contribution from inter\u00adatomic C\u22efH/H\u22efC contacts (Table\u00a04d). The presence of short inter\u00adatomic C\u22efC contacts are evident as the points near a rocket shape tip at de + di \u223c 3.2\u2005\u00c5 in the respective delineated fingerprint plot, Fig.\u00a010e), while the points corresponding \u03c0\u2013\u03c0 stacking between the imidazole and nitro\u00adbenzene rings are distributed about de = di\u00a0= 1.7\u2005\u00c5 in the plot. The small, i.e. 2.7%, contribution from C\u22efN/N\u22efC contacts to the surface is also due to these \u03c0\u2013\u03c0 stacking inter\u00adactions (delineated plot not shown). The contribution of 3.2% from C\u22efO/O\u22efC contacts is due to the presence of short inter\u00adatomic contacts involving nitro-O atoms, Table\u00a02de + di \u223c 3.2\u2005\u00c5 in the delineated plot of Fig.\u00a010f). The contribution from other inter\u00adatomic contacts to the surface summarized in Table\u00a04The overall two-dimensional fingerprint plot for (I)s Table\u00a04 to the Hs Table\u00a04 and the H-imidazol-2-yl)phenolate ligands in the literature ; complex (IV) has crystallographic twofold symmetry. The final structure, a copper(II) complex \u2013(VI) of 0.48, 0.53, 0.44, 0.37, 0.47 and 0.35, respectively.There are five crystal structures of copper complexes with related 2-In a typical procedure, benzil , ammonium acetate , 2-hy\u00addroxy-5-nitro\u00adbenzalaldehyde and copper(II) borate were ground in an agate mortar with a pestle. To this mixture, about 1.5\u2005g of dried silica gel was added and the reaction mixture was ground again for 30\u2005min. The whole reaction mixture was then transferred to a 100\u2005ml round-bottomed flask and heated at 130 \u00b0C with constant stirring for 4\u2005h. The reaction mixture was then extracted with dry acetone and dried over MgSOUiso(H) values set at 1.2Ueq(C). The O- and N-bound H atoms were located in a difference Fourier map but were refined with distance restraints of O\u2014H = 0.84\u2005\u00b1\u20050.01\u2005\u00c5 and N\u2014H = 0.88\u2005\u00b1\u20050.01\u2005\u00c5, respectively, and with Uiso(H) set at 1.5Ueq(O) or 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989019013720/hb7859sup1.cifCrystal structure: contains datablock(s) . DOI: 10.1107/S2056989019013720/hb7859Isup2.hklStructure factors: contains datablock(s) I. DOI: 1958158, 1958158CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the cation, the geometry around the Se atom is T-shaped, resulting from the coordination of Se by the C atom of the central aromatic ring and the N atoms of the benzimidazolyl moieties.In selenium} tetra\u00adkis\u00ad(seleno\u00adcyanato)\u00admercury, (C34H41N4Se)2[Hg(SeCN)4], the aryl\u00adselenenium cations, [C34H41N4Se]+, are linked through [Hg(SeCN)4]2\u2212 anions by C\u2014H\u22efN hydrogen bonds. In the cation, the geometry around the Se atom in the 5-tert-butyl-1,3-bis\u00ad(1-pentyl-1H-benzimidazol-2-yl)benzene scaffold is T-shaped, resulting from the coordination of Se by the C atom of the central aromatic ring and the N atoms of both of the benzimidazole moieties. The trans Se\u2014N bond lengths are almost equal [2.087\u2005(3) and 2.099\u2005(3)\u2005\u00c5] and the Se\u2014C bond length is 1.886\u2005(3)\u2005\u00c5. The N\u2014Se\u2014N angle is 159.29\u2005(11)\u00b0. The geometry around the HgII atom in the [Hg(SeCN)4]2\u2212 anion is distorted tetra\u00adhedral, with Se\u2014Hg\u2014Se angles ranging from 88.78\u2005(3) to 126.64\u2005(2)\u00b0. In [Hg(SeCN)4]2\u2212, the Hg\u2014Se bonds are unsymmetrical [2.5972\u2005(4) and 2.7242\u2005(5)\u2005\u00c5]. One of the pentyl substituents is disordered over two equivalent conformations, with occupancies of 0.852\u2005(8) and 0.148\u2005(8).In the title salt, bis\u00ad{[5- Some of these were investigated for their photoluminescence properties (Wang tert-but\u00adyl)-2,6-bis\u00ad(1-pentyl-1H-benzo[d]imidazol-2-yl)phen\u00adyl}(seleno\u00adcyanato)\u00admercury (3), mercury(II) chloride (1) was reacted with potassium seleno\u00adcyanate in 1,4-dioxane under reflux conditions. It was observed that, instead of the formation of the desired compound, the reaction leads to the isolation of an aryl\u00adselenenium(II) cation via auto-ionization 2 as by-products.In an attempt to synthesize {4-(2 reacts with an excess of KSeCN to form K2[Hg(SeCN)4] .HgCl2, crystallizes in the monoclinic space group C2/c. The asymmetric unit contains a selenenium cation along with half of a [Hg(SeCN)4]2\u2212 anion with the Hg atom located on a crystallographic twofold axis \u2005\u00c5, which is comparable with that found for a NCN pincer-based selenenium cation +[PF6]\u2212 C6H3Se]+[Br3]\u2212 (1.84\u2005\u00c5). The Se3\u2014N1 and Se3\u2014N2 bond lengths are almost equal [2.087\u2005(3) and 2.099\u2005(3)\u2005\u00c5]. The Se\u2014N distances are shorter than the sum of the van der Waals radii for Se and N [\u03a3rvdw 3.45\u2005\u00c5] and longer than the covalent radii [\u03a3rcov 1.91\u2005\u00c5] form a plane (r.m.s deviation for fitted atoms of 0.0182\u2005\u00c5) with the Se in this plane [deviation from the plane of 0.011\u2005(2)\u2005\u00c5].The title compound, is Fig.\u00a01. In the 4]2\u2212 acts as a bridging moiety between two selenenium cationic units. The Se3\u22efSe2 distance is 4.189\u2005(2)\u2005\u00c5 and the C1\u2014Se3\u22efSe2 angle is 163.40\u2005(9)\u00b0, which indicates that there is a weak secondary inter\u00adaction between the two different kinds of Se atoms in the cation and anion (Se3 and Se2). In the [Hg(SeCN)4]2\u2212 anion, two sets of Hg\u2014Se bonds exist. One set is shorter [2.5972\u2005(4)\u2005\u00c5] and the other set is longer [2.7242\u2005(5)\u2005\u00c5]. The Hg\u2013SeCN moieties are not linear, with Hg\u2014Se\u2014C angles of 101.31\u2005(14) and 101.43\u2005(11)\u00b0.In the anion, the mercury atom is coordinated by four seleno\u00adcyanate anions and the geometry around the mercury atom is distorted tetra\u00adhedral with Se\u2014Hg\u2014Se angles ranging from 88.78\u2005(3) to 126.64\u2005(2)\u00b0. The tetra\u00adcyano\u00adseleno\u00admercurate anion [Hg(SeCN)b-axis direction as shown in Fig.\u00a02A\u22efN1S and C18A\u2014H18C\u22efN2S inter\u00adactions ]selenenium cation. However, there have been several reports of structures containing [Hg(SeCN)1 in 1,4-dioxane (30\u2005ml) was added potassium seleno\u00adcyanate dissolved in MeOH. The reaction mixture was stirred for 6\u2005h under a nitro\u00adgen atmosphere and refluxed. The reaction mixture was filtered and the precipitate was washed with dioxane. Colourless prism-shaped crystals of 2 were obtained by layering a MeOH solution with diethyl ether at room temperature.To a solution of \u22121): 3059 (w), 2957 (s), 2931 (s), 2869 (s), 2124 , 1614 (m), 1464 (s), 1458 (s), 1440 (s), 1330 (w), 1288 (w), 1273 (w), 1154 (w), 1137 (w), 1011 (w), 892 (w), 746 (s). ESI\u2013MS: m/z calculated for C34H41N4Se: 585.2496. Found: 585.2552.Yield 11% ; m. p. turned blackish after 423\u2005K was reached. FT\u2013IR (KBr) (cmUiso(H) = xUeq(C), where x = 1.5 for methyl H atoms and 1.2 for all other C-bound H atoms. One of the pentyl substituents is disordered with an occupancy ratio of 0.852\u2005(8):0.148\u2005(8). It was refined as two equivalent conformations using SAME and SIMU instructions (SAME 0.01 and SIMU 0.01).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018006394/zl2726sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018006394/zl2726Isup2.hklStructure factors: contains datablock(s) I. DOI: 1839609CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The copper(II) atom shows a typical Jahn\u2013Teller distorted [4\u00a0+\u00a02] octa\u00adhedral coordination sphere. H-benzotriazol-1-yl)acetic acid and mono\u00adethano\u00adlamine with CuCl2\u00b72H2O resulted in the formation of the title complex, [Cu(C8H6N3O2)2(C2H7NO)2] or [Cu(BTA)2(MEA)2]. Its asymmetric unit comprises one BTA anion coordin\u00adating to the Cu2+ cation (site symmetry R22(8) graph-set motif. The mol\u00adecules are further linked by C\u2014H\u22ef\u03c0 inter\u00adactions involving the triazole rings and methyl\u00adene groups of MEA, thus generating an overall three-dimensional supra\u00admolecular framework.The reaction of 2-(1 The Cu2+ cation is located on a center of inversion. Its coordination polyhedron is a distorted N2O4 octa\u00adhedron formed by two oxygen atoms (O2) of the carb\u00adoxy groups of symmetry-related BTA anions, by two nitro\u00adgen atoms (N4) of two symmetry-related MEA ligands in the equatorial plane and by two O atoms (O3) of the same set of MEA ligands in the axial positions. The Cu\u2014O2 and Cu\u2014N4 bond lengths are 2.029\u2005(1) and 1.980\u2005(2)\u2005\u00c5, respectively, whereas the length of the axial Cu\u2014O3 bond is 2.492\u2005(2)\u2005\u00c5, typical for Jahn\u2013Teller distortions. The MEA ligand is neutral and acts as a bidentate N- and O-donor ligand and forms CuNC2O five-membered chelate rings which have a twist conformation; the O3\u2014C10\u2014C9\u2014N4 torsion angle is \u221260.3\u2005(3)\u00b0. The planar benzotriazole ring system (N1\u2013N3/C1\u2013C6: r.m.s. deviation = 0.0064\u2005\u00c5) is co-planar with the methyl carbon atom C7 [deviation from the plane of 0.158\u2005(2)\u2005\u00c5], whereas the carboxyl\u00adate group is nearly normal to this plane [88.0\u2005(2)\u00b0]. The difference of the C8\u2014O distances of the carboxyl\u00adate group (\u0394 = 0.036\u2005\u00c5) is due to the monodentate coordination, with the longer C\u2014O distance involving the coordinating O2 atom.The mol\u00adecular structure of The mol\u00adecular structure is stabilized by an intra\u00admolecular O3\u2014H3\u22efO1 hydrogen bond between the OH group of the MEA ligand and the non-coordinating carboxyl\u00adate O atom Fig.\u00a01.A\u22efO1iii, N4\u2014H4A\u22efO2ii and N4\u2014H4B\u22efO3ii hydrogen bonds between the amino function and carboxyl\u00adate/hy\u00addroxy O-atom acceptors generate a three-dimensional supra\u00admolecular framework.In the crystal structure of (I)s Table\u00a01, forming0) Fig.\u00a03. AdditioH-benzotriazol-1-yl)acetic acid and different metal cations in the CSD was slowly added an ethanol solution (5\u2005ml) containing MEA and HBTA under constant stirring. Blue crystals of the product were obtained by solvent evaporation at room temperature after one week. Yield: 70%. Elemental analysis: Calc. for C20H26CuN8O6 (538.04): C, 44.65; H, 4.87\u2005N, 20.83%. Found: C, 44.73; H, 4.93; N, 20.88%.To an aqueous solution (2.5\u2005ml) of CuClUiso(H) = 1.2Ueq(C). The positions of the O- and N bound H atoms were located from a difference-Fourier map and were refined with soft distance restraints, 0.82\u2005\u00c5 for the hydroxyl group and 0.95\u2005\u00c5 for the primary amine group.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019000744/wm5481sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019000744/wm5481Isup2.hklStructure factors: contains datablock(s) I. DOI: 1891272CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II ion lies on a crystallographic twofold axis and has distorted tetra\u00adhedral coordination geometry. Two weak C\u2014H\u22efS intra\u00admolecular hydrogen bonds exist between the bipyridyl and thiol groups. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds, forming a three-dimensional supra\u00admolecular architecture.The Zn 2N,N\u2032)bis\u00ad(2-meth\u00adoxy\u00adethyl xanthato-\u03baS)zinc(II), [Zn(C4H7O2S2)2(C10H8N2)], the ZnII ion is coordinated to two N atoms of the 2,2\u2032-bi\u00adpyridine ligand and two S atoms from two 2-meth\u00adoxy\u00adethyl xanthate ligands. The ZnII ion lies on a crystallographic twofold rotation axis and has distorted tetra\u00adhedral coordination geometry. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO hydrogen bonds, forming supramolecular chains propagating along the a-axis direction. Weak intra\u00admolecular C\u2014H\u22efS hydrogen bonds are also observed. The inter\u00admolecular contacts in the crystal were further analysed using Hirshfield surface analysis, which indicates that the most significant contacts are H\u22efH (36.3%), followed by S\u22efH/H\u22efS (24.7%), C\u22efH/H\u22efC (15.1%), O\u22efH/H\u22efO (14.4%), N\u22efH/H\u22efN (4.1%) and C\u22efC (2.9%).In the title compound, (2,2\u2032-bi\u00adpyridine-\u03ba ROCS2\u2212) have attracted the attention of scientific groups of researchers due to their diverse applications. Metal xanthates have been used as single-source precursors to metal sulfide materials and 2.295\u2005(2)\u2005\u00c5, respectively, whereas the bond angles around the central ZnII ion are in the range 78.7\u2005(3)\u2013126.64\u2005(10)\u00b0 . The C\u2014O bond lengths range from 1.346\u2005(8) to 1.453\u2005(8)\u2005\u00c5 although all of the C\u2014O bonds show single-bond character. In the {S2C} part of the xanthate ligands, the C1\u2014S1 distance is 1.727\u2005(7)\u2005\u00c5, which is typical of a single bond whereas the C1\u2014S2 distance of 1.652\u2005(7)\u2005\u00c5 is typical of a carbon-to-sulfur double bond. The C\u2014N and C\u2014C bond lengths in 2,2\u2032-bi\u00adpyridine are normal for 2-substituted pyridine derivatives (O-n-propyl\u00addithio\u00adcarbonato-S)zinc(II) zinc(II) and \u00adbis\u00ad(O-iso\u00adbutyl\u00adxan\u00adth\u00adato)zinc(II) . 2H2O in 2-meth\u00adoxy\u00adethanol, was added a hot solution of 2,2\u2032-bipy in 2-meth\u00adoxy\u00adethanol. A hot solution of potassium 2-meth\u00adoxy\u00adethylxanthate in 2-meth\u00adoxy\u00adethanol was added under stirring. Colourless crystals were formed after 30 minutes. The crystals were washed with small amounts of 2-meth\u00adoxy\u00adethanol and water and air-dried.To a hot solution of Zn(CHUiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) otherwise. The crystal was a weak diffractor (I/\u03c3 at 0.81 resolution was 5.1) and refinedas a two-component twin with HKLF 4 data (twin law \u22121 0 0 0\u00a0\u2212\u00a01 0 0 0\u00a0\u2212\u00a01) but this had little effect. The anisotropy of N1 was restrained with ISOR 0.01 0.02 in SHELXL I. DOI: 10.1107/S2056989019014968/lh5934Isup2.hklStructure factors: contains datablock(s) I. DOI: 1424075, 1424075CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit contains two independent mol\u00adecules and two water mol\u00adecules. The central parts of both the mol\u00adecules are twisted as both mol\u00adecules are bent at both the S and N atoms. The crystal structure features N\u2014H\u22efN, N\u2014H\u22efO, C\u2014H\u22efO and O\u2014H\u22efO inter\u00admolecular inter\u00adactions. Two-dimensional fingerprint plots show that the largest contributions to the crystal stability come from O\u22efH/H\u22efO and H\u22efH inter\u00adactions. 9H8ClN3O3S2\u00b7H2O, consists of two independent mol\u00adecules and two water mol\u00adecules. The central parts of the mol\u00adecules are twisted as both the mol\u00adecules are bent at both the S and N atoms. In the crystal, N\u2014H\u22efN, N\u2014H\u22efO, C\u2014H\u22efO and O\u2014H\u22efO hydrogen-bonding inter\u00adactions connect the mol\u00adecules, forming layers parallel to the ab plane. Two-dimensional fingerprint plots associated with the Hirshfeld surface show that the largest contributions to the crystal packing come from O\u22efH/H\u22efO (32.9%) and H\u22efH (22.6%) inter\u00adactions.The asymmetric unit of the title thia\u00adzole derivative containing a sulfonyl\u00adhydrazinic moiety, C The sulfonyl\u00adhydrazide bond exists in the synclinal conformation preferred by aromatic sulfonamides benz\u00adene\u00adsulfono\u00adhydrazide -methyl 2-[(Z)-4-oxo-2-(2-tosyl\u00adhydrazono)thia\u00adzolidin-5-yl\u00adidene]acetate and (Z)-methyl-2-[(Z)-2-(ethyl\u00adimino)-4-oxo-3-(phenyl\u00adamino)\u00adthia\u00adzolidin-5-yl\u00adidene]acetate phen\u00adyl]-1,3-thia\u00adzol-2(3H)-yl\u00adidene}benzene\u00adsulfono\u00adhydrazide and related fingerprint plots were generated using CrystalExplorer17.5 /rivdw\u00a0+\u00a0(de\u00a0\u2212\u00a0revdw)/revdw with di + de \u223c 1.9\u2005\u00c5. The presence of water mol\u00adecules in the unit cell provides the largest contribution to the stability of the crystal packing. The next largest contrib\u00adutor is from H\u22efH inter\u00adactions, which contribute 22.6%. A single sharp spike can be seen in the middle region of the plot, at di = de = 0.9\u2005\u00c5 . The N\u22efH contacts, which refer to N\u2014H\u22efN inter\u00adactions, contribute 5.3% to the surface. Two sharp spikes having di + de = 1.8\u2005\u00c5 are observed. The C\u22efH contacts contribute 5.9% to the Hirshfeld surface, featuring a wide region with di + de = 3.1\u2005\u00c5 . The different inter\u00adatomic contacts and percentage contributions to the Hirshfeld surface are Cl\u22efH/H\u22efCl (8.3%), S\u22efH/H\u22efS (6.1%), Cl\u22efO/O\u22efCl (3.0%), Cl\u22efC/C\u22efCl (2.4%), S\u22efO/O\u22efS (1.7%), and C\u22efO/O\u22efC (1.6%) as depicted in the fingerprint plots .In order to explore the role of weak inter\u00admolecular inter\u00adactions in the crystal packing, Hirshfeld surfaces benzene-1-sulfono\u00adhydrazide was prepared by adding 4-chloro benzene\u00adsulfonyl chloride (0.02\u2005mol) under stirring to a solution of thio\u00adsemicarbazide (0.02\u2005mol) in 5% aqueous NaOH solution (20\u2005ml). The reaction mixture was stirred at room temperature for 1\u2005h, then diluted twofold with water and neutralized with glacial acetic acid. The solid 2-(4-chloro\u00adbenzene-1-sulfon\u00adyl)hydrazine-1-carbo\u00adthio\u00adamide (A) obtained was crystallized from acetic acid. Mono\u00adchloro\u00adacetic acid (0.01\u2005mol) and anhydrous sodium acetate (0.04\u2005mol) were added to A (0.01\u2005mol) in glacial acetic acid. The reaction mixture was refluxed for 8\u201310\u2005h and the completion of the reaction was checked by TLC. The reaction mixture was then poured into cold water. The resulted precipitate of the title compound was separated by vacuum filtration. Prismatic colourless single crystals of the title compound were grown from a mixture of aceto\u00adnitrile-DMF (5:1 v/v) by slow evaporation of the solvent. The purity of the compound was checked by TLC and characterized by IR spectroscopy. The characteristic IR absorptions observed at 3095.9, 1639.5, 1458.7, 1343.2, 1139.4, and 1215.7\u2005cm\u22121 correspond to N\u2014H, C=O, C=N, S=O asymmetric and symmetric, and C\u2014S absorptions, respectively. The 1H and 13C spectra of the title compound are as follows: 1H ; \u03b4 3.45 , 7.68\u20137.86 , 10.01 , 11.96 . 13C NMR ; \u03b4 36.8, 128.4, 129.1, 131.1,132.5, 133.9, 137.2, 165.4, 185.5.4-Chloro-Ueq of the parent atom.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018013658/rz5243sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018013658/rz5243Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018013658/rz5243Isup3.cmlSupporting information file. DOI: 1869597CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N(4)-morpholino\u00adthio\u00adsemicarbazone forms sheets parallel to (002) and consisting of two parallel chains running in the a-axis direction and formed by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds.In the crystalline state, salicyl\u00adaldehyde N-{[(2-hy\u00addroxy\u00adphen\u00adyl)methyl\u00adidene]amino}\u00admorpholine-4-carbo\u00adthio\u00adamide), C12H15N3O2S, was prev\u00adiously determined \u00b0. In the crystal, the mol\u00adecules are connected into chains by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, which combine to generate sheets lying parallel to (002). The most prominent contribution to the surface contacts are H\u22efH contacts (51.6%), as concluded from a Hirshfeld surface analysis.The structure of the title compound (systematic name: Cisplatin is well known as an effective therapy to prohibit the proliferation of tumor cells -substituted thio\u00adsemicarbazide. Many reports have demonstrated that N(4)-aromatic or heterocyclic substituted thio\u00adsemicarbazides are biologically more active than thio\u00adsemicarbazones without substituted groups -morpholino\u00adthio\u00adsemicarbazone was published previously -morpholino\u00adthio\u00adsemicarbazone (3) together with its structural characteristics and crystal structure redetermination using present-day technology.The crystal and mol\u00adecular structure of salicyl\u00adaldehyde Pna21 with one mol\u00adecule in the asymmetric unit \u00b0], which gives rise to an intra\u00admolecular O18\u2014H18\u22efN10 hydrogen bond with an ap conformation].The title compound crystallizes in the ortho\u00adrhom\u00adbic space group it Fig.\u00a01. The N9\u2014f Table\u00a01. The plaQ = 0.554\u2005(3)\u2005\u00c5, \u03b8 = 173.2\u2005(3)\u00b0 and \u03c6 = 214\u2005(3)\u00b0] with the thio\u00adsemicarbazone function in an equatorial position. The plane of the phenyl ring forms a dihedral angle of 43.44\u2005(17)\u00b0 with the best plane through the morpholino ring. A second intra\u00admolecular C6\u2014H6A\u22efS8 inter\u00adaction is observed (Table\u00a01The morpholino ring adopts a chair conformation .A search of the Cambridge Structural Database and \u2212175.97\u2005(15)\u00b0] and \u03c42 [166.51\u2005(16) and \u2212174.99\u2005(16)\u00b0] are similar to those observed for the title compound. An intra\u00admolecular hydrogen bond similar to O18\u2014H18\u22efN10 is also observed.The most similar compound present in the CSD is the 2-hy\u00addroxy\u00adnaphthaldehyde-based thio\u00adsemicarbazone is given in Fig.\u00a07The reaction scheme for the synthesis of (Synthesis of 2-((morpholine-4-carbono\u00adthio\u00adyl)thio)\u00adacetic acid (1):A mixture consisting of carbon di\u00adsulfide (0.2\u2005mol) and concentrated ammonia (25\u2005mL) was stirred to form a homogeneous solution at 278\u2005K. Then, morpholine (0.2\u2005mol) was added dropwise to this solution. The yellow solid that separated from the solution was filtered off and immediately dissolved in deionized water (300\u2005mL) at room temperature to generate a yellow solution. Sodium chloro\u00adacetate (0.2\u2005mol) was added to this solution and the reaction mixture maintained for 6\u2005h at room temperature. The yellowish solution was acidified with concentrated hydro\u00adchloric acid and the resulting white precipitate was filtered off and recrystallized from ethanol.Synthesis of N(4)-morpholino\u00adthio\u00adsemicarbazide (2):1) (50\u2005mmol), deionized water (10\u2005mL) and hydrazine hydrate (25\u2005mL) was refluxed for 30 minutes at 353\u2005K. The white solid which precipitated from the transparent solution was filtered off and recrystallized from ethanol to give (2).A mixture composed of -morpholino\u00adthio\u00adsemicarbazone (3):2) in hot ethanol, the solution was added to an equivalent amount of salicyl\u00adaldehyde. The final solution was refluxed at 353\u2005K for 2\u2005h in the presence of acetic acid as a catalyst. The resulting solution was gradually reduced in volume at room temperature overnight. The needle-shaped crystals that formed were filtered off and recrystallized from ethanol to give (3) in the form of transparent crystals (yield 60%), m.p. 461\u2013463\u2005K. FT\u2013IR (cm\u22121): 3436 (O\u2014H), 3279 (N\u2014H), 1617 (CAr\u2014H), 1540 (C=N), 1061 (N\u2014N), 1348 and 959 (C=S). 1H NMR : 3.67 ; 3.92 ; 6.90 ; 7.28 ; 7.41 ; 8.47 ; 11.49 ; 11.55 . 13C NMR : 49.4 (C2 and C6), 66.2 (C3 and C5), 117.0 (C14), 118.9 (C12), 119.5 (C16), 130.4 (C17), 131.3 (C15), 146.9 (C13), 157.6 (C11), 180.1 (C7). UV\u2013Vis : 200 (\u03c0\u2192\u03c0*); 300 and 350 (n\u2192\u03c0*).After dissolving (Uiso(H) values 1.2Ueq of the parent atoms, with C\u2014H distances of 0.93 and 0.97\u2005\u00c5 (CH2). In the final cycles of refinement, 4 outliers were omitted.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019011812/mw2147sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019011812/mw2147Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019011812/mw2147Isup3.cmlSupporting information file. DOI: 1949697CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "EVs). Three mouse melanoma B16 variants with distinct metastatic potentials show similar gene expression levels and enzymatic activities of glycosyltransferases involved in N\u2010glycosylation. All melanoma variants and EVs have nearly identical profiles of de\u2010sialylated N\u2010glycans. The major de\u2010sialylated N\u2010glycan structures of cells and EVs are core\u2010fucosylated, tetra\u2010antennary N\u2010glycans with \u03b21,6\u2010N\u2010acetylglucosamine branches. A few N\u2010glycans are extended by N\u2010acetyllactosamine repeats. Sialylation of these N\u2010glycans may generate cell\u2010type\u2010specific N\u2010glycomes on EVs. Taken together, melanoma\u2010derived EVs show high expression of tumor\u2010associated N\u2010glycans, and the core structure profile is inherited during multiple selection cycles of B16 melanomas and from tumor cells to EVs.We investigated the correlation between metastatic behaviors of tumor cells and asparagine\u2010linked glycosylation (N\u2010glycosylation) of tumor\u2010derived extracellular vesicles ( EVs, extracellular vesiclesPA, 2\u2010aminopyridineN\u2010acetylglucosamine (GlcNAc) may be fucosylated (core fucosylation) Most cell surface and secretory proteins are modified by asparagine\u2010linked glycans (N\u2010glycans) in\u00a0vivo and in\u00a0vitro selections have been performed to obtain mouse B16 malignant melanoma variants with distinct metastatic potentials. The B16\u2010F1 (poorly lung\u2010colonizing) and B16\u2010F10 (highly lung\u2010colonizing) variants are selected for their lung colonization abilities following intravenous injection of the parent B16 cells in\u00a0vitro, resulting in a highly spontaneous metastatic variant Blood\u2010borne tumor metastasis is a complex process involving tumor cell invasion into normal tissues, intravasation to the circulation, extravasation, and colonization at distant organs Tumor cells secrete small vesicles, termed extracellular vesicles (EVs), that contain various cargo molecules including nucleic acids, soluble proteins, and membrane proteins in\u00a0vivo and in\u00a0vitro selection cycles of B16 variants and is copied from tumor cells to their EVs. It was suggested that sialylation of these N\u2010glycans probably generates cell\u2010type\u2010specific N\u2010glycome on EVs. This study establishes the N\u2010glycosylation landscapes of B16 variants and their EVs, and indicates that the bulk N\u2010glycosylation of melanoma EVs does not reflect the metastatic potentials of their secreting tumor cells in the B16 model.In the present study, we investigated the correlation between metastatic potentials of three B16 variants and N\u2010glycosylation of their EVs by characterizing N\u2010glycan structures and gene expression, as well as enzymatic activity, of glycosyltransferases involved in N\u2010glycosylation. The results demonstrate that the core structure profile of N\u2010glycans is inherited at the genetic level during multiple \u22121 penicillin, and 100\u00a0\u03bcg\u00b7mL\u22121 streptomycin at 37\u00a0\u00b0C in a 5% CO2 atmosphere.Mouse B16\u2010F1 and B16\u2010F10 melanoma cells were purchased from the American Type Culture Collection . B16\u2010BL6 cells were purchased from RIKEN Bioresource Center. All cell lines were maintained in complete Dulbecco's modified Eagle's medium containing 10% fetal bovine serum, 100\u00a0U\u00b7mL6 cells/10\u2010cm dish) were cultured for 24\u00a0h, washed twice with PBS, and incubated for 48\u00a0h in EV\u2010depleted medium. The conditioned medium was sequentially centrifuged at 130\u00a0g for 5\u00a0min, 20\u00a0000\u00a0g for 20\u00a0min, and 100\u00a0000\u00a0g for 70\u00a0min. The final pellet containing EVs was washed once with PBS and measured for its protein concentration with a BCA protein assay kit , using bovine serum albumin as an external standard.Extracellular vesicles were prepared as described m Tris\u2010HCl (pH 7.4), 150\u00a0mm NaCl, 1% Triton X\u2010100, 1\u00a0mm EDTA, and protease inhibitor cocktail . The homogenate was centrifuged at 20\u00a0000\u00a0g for 10\u00a0min and the supernatant was recovered as the whole cell lysate.Cells were lysed for 15\u00a0min at 0\u00a0\u00b0C in lysis buffer comprising 10\u00a0mN4\u2010(N\u2010acetyl\u2010\u03b2\u2010glucosaminyl)asparagine amidase F (PNGase F) according to the manufacturer's instructions. The released N\u2010glycans were purified and fluorescently labeled with 2\u2010aminopyridine (PA) using a BlotGlyco Kit according to the manufacturer's instructions.N\u2010glycans were released from EVs and whole cell lysates by treatment with peptide\u2010Arthrobacter ureafaciens in 10\u00a0mm sodium acetate buffer (pH 5.5) for 16\u00a0h at 37\u00a0\u00b0C. The sialidase\u2010digested samples and the neutral fraction were desalted on PD\u201010 desalting columns and subjected to reversed\u2010phase HPLC as described \u22121; Takara), as described PA\u2010labeled N\u2010glycans were initially separated into one neutral fraction and six sialidase\u2010sensitive fractions by anion\u2010exchange HPLC, as described PA\u2010labeled N\u2010glycans fractionated by reversed\u2010phase HPLC were subjected to matrix\u2010assisted laser desorption/ionization time\u2010of\u2010flight mass spectrometric analysis. PA\u2010glycans (1\u00a0\u03bcL) were mixed with 1\u00a0\u03bcL of 2,5\u2010dihydroxybenzoic acid (10\u00a0mg/mL in 50% acetonitrile/0.1% trifluoroacetic acid) on a target plate. After evaporation to dryness, spectra were obtained in the positive mode using an Autoflex mass spectrometer operated in the reflector mode.http://www.glycoanalysis.info/galaxy2/ENG/index.jsp), using the masses and glucose units of individual glycans, the latter of which were calculated by the elution positions of PA\u2010glucose oligomers (degree of polymerization\u00a0=\u00a03\u201322) in the reversed\u2010phase and size\u2010fractionation chromatography The structures of PA\u2010labeled N\u2010glycans from melanoma EVs were determined by reference to a database, glycoanalysis by the three axes of MS and chromatography (GALAXY) Samples (10\u00a0\u03bcg protein) were denatured for 3\u00a0min at 100\u00a0\u00b0C, separated by SDS/PAGE, and analyzed by western blotting using an anti\u2010CD81 antibody .6 cells/10\u2010cm dish) that had been cultivated for 48\u00a0h using TRI Reagent in accordance with the manufacturer's protocol, followed by treatment with DNase (Qiagen) and purification using RNeasy Mini kit . 1\u00a0\u03bcg total RNA was reverse\u2010transcribed using an R2 First Strand Kit (Qiagen) in a 40\u2010\u03bcL reaction mixture and then diluted with 182\u00a0\u03bcL RNase\u2010free water. The cDNA thus obtained was mixed with 2.7\u00a0mL RT2 SYBR Green qPCR Mastermix (Qiagen) and 2.496\u00a0mL water, and 25\u00a0\u03bcL of the mixture was applied to each well of a 96\u2010well plate that contained specific primers for glycotransferase mRNAs (Qiagen). Two types of 96\u2010well plates were used for quantification of 144 glycosyltransferase genes; a commercially available plate for Mouse Glycosylation (Cat. No. 330231 PAMM\u2010046ZA) covering 84 glyco\u2010related genes, and a 96\u2010Well Custom PCR Array in which we manually selected 86 glycosyltransferase genes, seven house keeping genes and three primers for monitoring genomic contamination, cDNA synthesis and PCR reaction. cDNAs were amplified and analyzed using an ABI PRISM 7900HT thermocycler according to the RT2 Profiler PCR Array Handbook (Qiagen). The abundance of glycotransferase mRNAs relative to that of housekeeping genes was calculated using the \u0394Ct method. The values in Tables\u00a0Real\u2010time PCR for glycosyltransferases was performed as described previously m MES pH 6.2, 10\u00a0mm MnCl2, 10\u00a0mm UDP\u2010Gal at 37\u00a0\u00b0C overnight. The enzymes were mixed with various concentrations of a fluorescence\u2010labeled acceptor substrate equipped with an ODS column Enzymatic activity of mannosyl \u2010glycoprotein \u03b21,4\u2010GlcNAc transferase (Mgat3), \u03b1\u20101,3\u2010mannosyl\u2010glycoprotein 4\u2010\u03b2\u2010GlcNAc transferase (Mgat4), \u03b1\u20101,6\u2010mannosylglycoprotein 6\u2010\u03b2\u2010GlcNAc transferase (Mgat5), fucosyltransferase 8 (Fut8), \u03b12,3\u2010sialyltransferase and \u03b12,6\u2010sialyltransferase were measured as described previously cDNAs encoding the catalytic regions of human MGAT3, mouse Magt4a, human MGAT5, human FUT8, human ST3GAL4 and human ST6GAL1 were amplified by PCR with primers: human MGAT3, 5\u2032CGGAATTCGAGCCAGGAGGCCCTGACCT3\u2032 and 5\u2032CTAGACTTCCGCCTCGTCCA3\u2032; mouse Mgat4a, 5\u2032CGGAGCTAAACACCATTGTC3\u2032 and 5\u2032GTGCTCGAGTCTAAGCAGATCAACTGGTG3\u2032; human MGAT5, 5\u2032CTACAGCTGTTCCCAGCTTG3\u2032 and 5\u2032AGGCTCGAGCTATAGGCAGTCTTTGCAGA3\u2032; human FUT8, 5\u2032CGGAATTCCGGATACCAGAAGGCCCTAT3\u2032 and 5\u2032TTATTTCTCAGCCTCAGGAT3\u2032; human ST3GAL4, 5\u2032GAGGAATTCGAGCCGTGCCTCCAGGGTGA3\u2032 and 5\u2032TTGCTCGAGTCAGAAGGACGTGAGGTTCTTG3\u2032; and human ST6GAL1, 5\u2032GTGGAATTCAAGGACAGCTCTTCCAAAAA3\u2032 and 5\u2032AAACTCGAGTTAGCAGTGAATGGTCCGGAA3\u2032.2+\u2010column and desalted with NAP\u20105 gel filtration column .The PCR products were inserted into pcDNA\u2010IH as described previously Stt3a and Stt3b), protein quality control in the ER (Uggt1 and Uggt2), GlcNAc branch formation , lysosome targeting (Gnptab and Gnptg), galactosyltransferases , polylactosamine formation (B3gnt2 and B3gnt4), core fucosylation (Fut8), \u03b12,3\u2010sialyltransferases and \u03b12,6\u2010sialyltransferase . The gene expression pattern of B16\u2010F1 and B16\u2010BL6 cells was similar to that of B16\u2010F10 cells, except that B4galt2 was undetectable in B16\u2010BL6 cells. We hardly detected expression of Mgat3 (bisecting GlcNAc formation), as well as Fut1\u20107 and Fut9 , in all three variants. In comparison of cells with different lung colonization ability (B16\u2010F1 and B16\u2010F10), B16\u2010F1 cells showed higher expression of Mgat5 (1.5\u2010fold), B4galt2 (3\u2010fold), B3gnt2 (1.5\u2010fold), St3gal4 (2.0\u2010fold) and St6gal1 (1.4\u2010fold) than B16\u2010F10 cells. In contrast, B16\u2010F1 cells expressed lower levels of Stt3a (0.8\u2010fold), B3galt2 (0.7\u2010fold) and Fut8 (0.8\u2010fold) than B16\u2010F10 cells. In comparison of cells with different invasiveness (B16\u2010F10 and B16\u2010BL6), B16\u2010BL6 cells showed higher levels of Uggt2 (1.8\u2010fold), Mgat4b (1.4\u2010fold), B4galt1 (1.2\u2010fold), B3galt2 (1.2\u2010fold), St3gal4 (1.3\u2010fold) and St6gal1 (1.9\u2010fold) than B16\u2010F10 cells. In contrast, B16\u2010BL6 cells showed lower expression of Uggt1 (0.7\u2010fold), Mgat4a (0.7\u2010fold) and Mgat5 (0.8\u2010fold) than B16\u2010F10 cells, suggesting that low expression of Uggt1 and Mgat4a is compensated by high expression of Uggt2 and Mgat4b in B16\u2010BL6 cells. Together, these results indicate that B16 variants with distinct metastatic potential have unique glycosyltransferase gene expression patterns. However, overall N\u2010glycosylation\u2010related gene profiles are inherited during multiple selection cycles of B16 variants.The cellular N\u2010glycome is regulated by multiple factors including the expression profiles of glycosyltransferase genes Mgat3 gene was rarely expressed in all three B16 variants Among the three B16 variants, the B16\u2010F10 variant is positioned in the middle of the selection cycles N\u2010acetyllactosamine repeats (polylactosamine), which can serve as ligands for galactose\u2010binding lectins As the sialylation of N\u2010glycans generated considerable structural heterogeneity, de\u2010sialylated N\u2010glycans were subjected to further structural analyses. Neutral and de\u2010sialylated Sia 1\u20106 fractions from anion\u2010exchange HPLC were further fractionated by reversed\u2010phase HPLC Fig.\u00a0C and subTo investigate whether characteristic core N\u2010glycan structures were enriched in EVs, we compared reversed\u2010phase HPLC profiles of de\u2010sialylated complex\u2010type glycans between F10\u2010EVs and B16\u2010F10 cells. For the comparison, we chose eight major peaks that contained nine N\u2010glycans Fig.\u00a0A and fouNext, we investigated whether the N\u2010glycosylation profiles of EVs differed among the B16 variants. Anion\u2010exchange HPLC analysis of sialylated glycans obtained from F1\u2010EVs, F10\u2010EVs, and BL6\u2010EVs revealed that although the number of sialic acids present on N\u2010glycans was similar among the three variants, the Sia 3 fraction showed distinct elution patterns , the Kodama Memorial Fund for Medical Research (YH), and MEXT/JSPS Grants\u2010in\u2010Aid for Scientific Research (JP17H06414 to HY and 17K07356 to YK).Fig.\u00a0S1. Relative gene expression levels of 144 glycosyltransferases in B16 variants.Click here for additional data file.Fig.\u00a0S2. Comparative analysis of sialylated N\u2010glycans from B16\u2010F10 cells and F10\u2010EVs.Click here for additional data file.Fig.\u00a0S3. Relative amounts of sialylated N\u2010glycans in the Sia 1\u20106 fractions of B16\u2010F10 cells and F10\u2010EVs.Click here for additional data file.Table\u00a0S1. mRNA abundances of glycosyltransferases relative to the mean abundance of four housekeeping genes in B16 variants.Table\u00a0S2. Structural analysis of N\u2010glycans expressed on EVs from B16\u2010F10 cells.Click here for additional data file."} +{"text": "In the crystal structure, inter\u00admolecular N\u2014H\u22efN hydrogen bonds link the mol\u00adecules into infinite ribbons extending along the [100] direction. 12H10N6, at 100\u2005K has monoclinic (P21/n) symmetry. Crystals were obtained as a yellow solid by reduction of 3,6-bis\u00ad(pyridin-2-yl)-1,2,4,5-tetra\u00adzine. The structure displays inter\u00admolecular hydrogen bonding of the N\u2014H\u22efN type, ordering mol\u00adecules into infinite ribbons extending along the [100] direction.The structure of the title compound, C The substitution of four nitro\u00adgen atoms in a six-membered benzene-like ring results in strong \u03c0-electron deficiency and concentration of negative charge on the heteroatoms. As a result of these properties, s-tetra\u00adzines are used in organic synthesis in ethanol solution -1,4-di\u00adhydro-1,2,4,5-tetra\u00adzine (I)on Fig.\u00a01.P21/n. The atomic labelling scheme is shown in Fig.\u00a02Compound (I)et al., 1987A) ring are of almost equal length, being 1.4285\u2005(15) and 1.4306\u2005(16)\u2005\u00c5. The C6\u2014N1 and C3\u2014N4 [1.3953\u2005(17) and 1.4051\u2005(17)\u2005\u00c5] bond lengths are longer than those for C6\u2014N5 and C3\u2014N2 , respectively. This is the result of the protonation of the N1 and N4 atoms. The C\u2014N bond lengths in the B and C rings are comparable within 3\u03c3, varying from 1.3384\u2005(18)\u2005\u00c5 to 1.3416\u2005(17)\u2005\u00c5.The C\u2014C bond lengths are within the expected values known for aromatic systems (Allen A) shows a boat conformation with pseudo-symmetry mirror planes passing through bonds N2\u2014C3 and N5\u2014C6 [\u0394Cs = 1.30\u2005(16)\u00b0] and atoms N1, N4 [\u0394Cs = 2.00\u2005(14)\u00b0]. In this conformation, hydrogen atoms are located in the equatorial positions of the ring and the N\u2014H bonds are directed to the bottom of the boat (compare torsion angles in Table\u00a01B and C) are not to parallel to each other. The dihedral angles between these rings and central tetra\u00adzine ring are 22.43\u2005(7)\u00b0 (A and B) and 25.71\u2005(6)\u00b0 (A and C). The dihedral angle between rings B and C is 27.13\u2005(7)\u00b0. The overall mol\u00adecular structure could be recognized as a butterfly-like conformation as shown in Fig.\u00a03The central tetra\u00adzine ring (R22(6) ring motif = 3.2418\u2005(18)\u2005\u00c5 and C34\u22efC61 = 3.3334\u2005(19)\u2005\u00c5], as shown in Fig.\u00a05The crystal packing of (I)et al., 2016et al., 1998A search of the Cambridge Structure Database were mixed in ethanol (4\u2005ml). The resulting solution was warmed to 343\u2005K and then kept at room temperature. Within two weeks, after slow evaporation of the solvent, two kinds of crystal were obtained in a crystallizer. X-ray studies confirmed that the pink crystals were of the known structure (II), while the yellow crystals were identified as being of a previously unreported structure, i.e. (I)Crystals suitable for X-ray measurements were obtained from a commercially available reagent and used without further purification. 0.5\u2005mmol of 3,6-bis\u00ad(pyridin-2-yl)-1,2,4,5-tetra\u00adzine and 0.5\u2005mmol of 2-mercapto\u00adpyridine Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901801753X/ff2157sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901801753X/ff2157Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901801753X/ff2157Isup3.cmlSupporting information file. DOI: 1884403CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "One of these molecules exhibits excellent gelation behaviors in water, and the resultant hydrogels are found to display self-healing properties. Interestingly, the elasticity and strength of the resulting gel can be tuned by the addition of different kinds of Hofmeister salts. The gel formation mechanism was proposed based on the analysis of FT-IR,1HNMR, and XRD, indicating that the main driving force for the self-assembly was the \u03c0-\u03c0 stacking of the benzene rings in the aqueous solution system. Overall, our research provides an efficient approach for facilely tuning the properties of the d-gluconic acetal-based hydrogel.Physical gelation behaviors of a series of The online version of this article (10.1186/s11671-018-2823-8) contains supplementary material, which is available to authorized users. Gels composed of low molecular mass gelators (LMMGs) can be obtained from the self-assembly of LMMGs via supramolecular interactions such as hydrogen bonding, van der Waals interactions, \u03c0-\u03c0 stacking, and so on \u201313. Over4)2SO4 solutions at appropriate concentrations. However, it has only recently been appreciated in supramolecular chemistry and, more specifically, LWMGs-based hydrogels. For example, Mocerino et al. D2O, the signals of C\u2013H protons on the benzene ring of the G1 mixed with kosmotropes anions moved upfield, which indicated that the \u03c0-\u03c0 stacking effects between benzene rings were enhanced. Correspondingly after the addition of I\u2212, ClO42\u2212, and SCN\u2212, the peaks of C\u2013H protons shifted to 7.993, 8.007, and 8.130\u00a0ppm, respectively. Compared with G1 in pure [d6]D2O, the signals of C\u2013H protons on the benzene ring of the G1 mixed with chaotropes anions moved downfield, which suggested that the \u03c0-\u03c0 stacking effects between benzene rings were weakened. In Fig.\u00a0\u2212, Br\u2212, and NO3\u2212), the signals of C\u2013H protons on the benzene ring shifted slightly, indicating that the \u03c0-\u03c0 stacking effects between benzene rings were not influenced obviously.1H NMR spectra of the gelator G1 (2.5% . The result was consistent with the macro performance of the gelators (the changing trends of CGC and Tgel). It also reveals that the main driving force for the self-assembly is the \u03c0-\u03c0 stacking force of the benzene rings in the aqueous solution system.From kosmotropes to chaotropes, H-shift on the benzene ring gradually moved toward the low field and the \u03c0-\u03c0 stacking effect between benzene rings was gradually weakened42\u2212 , 2\u03b8\u2009=\u200928.262 (d\u2009=\u20090.31551\u00a0nm), 2\u03b8\u2009=\u200934.071 (d\u2009=\u20090.26292\u00a0nm), and 2\u03b8\u2009=\u200938.843 (d\u2009=\u20090.23165), and the ratio of d-spacing values is about 1:1/\u221a2:1:\u221a3 indicating that the self-assembly of the G1 xerogel from water is composed of hexagonal closs packing possibly [d\u2009=\u20090.38002\u00a0nm), and d\u2009=\u20090.38\u00a0nm is the characteristic of \u03c0-\u03c0 stacking force of the benzene rings. It reveals that the main driving force for the self-assembly is the \u03c0-\u03c0 stacking force of the benzene rings in the aqueous solution system [2SO4 aqueous solution (concentration is 0.5\u00a0M) exhibited four diffraction peaks centered at 2\u03b8\u2009=\u200919.084 (d\u2009=\u20090.46468\u00a0nm), 2\u03b8\u2009=\u200928.075 (d\u2009=\u20090.31757\u00a0nm), 2\u03b8\u2009=\u200933.901 (d\u2009=\u20090.26421\u00a0nm), and 2\u03b8\u2009=\u200938.683 (d\u2009=\u20090.23257\u00a0nm), and it proved that the addition of SO42\u2212 did not affect packing modes of the gelator molecules.While patterns of the G1 xerogel from NaSCN aqueous solution (concentration is 0.5\u00a0M) exhibited diffraction peaks centered at 2\u03b8\u2009=\u200926.184 (d\u2009=\u20090.34006\u00a0nm), 2\u03b8\u2009=\u200930.263 (d\u2009=\u20090.29508\u00a0nm), 2\u03b8\u2009=\u200938.904 (d\u2009=\u20090.23130\u00a0nm), and 2\u03b8\u2009=\u200938.683 (d\u2009=\u20090.23257\u00a0nm), and patterns of the G1 xerogel from aqueous solution exhibited four diffraction peaks centered 2\u03b8\u2009=\u200923.076 (d\u2009=\u20090.38511\u00a0nm), 2\u03b8\u2009=\u200929.563 (d\u2009=\u20090.30191\u00a0nm), 2\u03b8\u2009=\u200932.101 (d\u2009=\u20090.27860\u00a0nm), and 2\u03b8\u2009=\u200939.165 (d\u2009=\u20090.22982\u00a0nm). The ratio of d-spacing values were all 1:1/\u221a2:1:\u221a3 indicating that the self-assembly of the G1 xerogel from water is mainly composed of hexagonal closs packing [To explore the possible packing modes of the gelator molecules in water with the addition of Hofmeister salts in particular, wide-angle XRD (WXRD) of the G1 xerogels were examined. As shown in Fig.\u00a0possibly . In addin system , 78. In Furthermore, the energy-minimized structure of G1 in in present of Hofmeister anions (concentration is 0.5\u00a0M): (a) Na2SO4, (b) Na2S2O3, (c) Na2HPO4, (d) NaF, (e) H2O, (f) NaCl, (g) NaBr, (h) NaNO3, (i) NaI, (j) NaClO4, (k) NaSCN at 25\u2009\u00b0C, demonstrating high G\u2019values , flowing point . The step-strain measurement shows the recovery ratios of G\u2019 after the first cycle . Figure S2. (a) Frequency sweep of hydrogels from G1 with Hofmeister salts (concentration is 0.5\u2009M) with a fixed strain (0.1%) at 25\u00a0\u00b0C; (b) Rheological data under oscillatory stress experiment on hydrogels from G1 with Hofmeister salts (concentration is 0.5\u2009M) with a fixed frequency (1\u2009Hz) at 25\u00a0\u00b0C; (c) Time scan tests under alternating strain of 0.1% and 100% of G1 with Hofmeister salts (concentration is 0.5\u00a0M) with a fixed frequency at 1\u2009Hz at 25\u00a0\u00b0C. Figure S3. SEM images of G1 xerogel obtained from hydrogel (2.5% w/v) in present of Na2SO4 aqueous solution(concentration is 0.5\u00a0M); (b) SEM images of G1 xerogel obtained from hydrogel (2.5% w/v); (c) SEM images of G1 xerogel obtained from hydrogel (2.5% w/v) in present of NaCl aqueous solution(concentration is 0.5\u00a0M); (d) SEM images of G1 xerogel obtained from hydrogel (2.5% w/v) in presence of NaSCN aqueous solution (concentration is 0.5\u2009M). Figure S4. The energy-minimized mode of G1. The length of molecular PG16 is 14.6\u2009\u00c5. (DOCX 3013 kb)"} +{"text": "The title compound is built up from a planar quinoxalinone ring system linked through a methyl\u00adene bridge to a 1,2,3-triazole ring, which is inclined by 67.09\u2005(4)\u00b0 to the quinoxalinone ring plane. 16H19N5O, is built up from a planar quinoxalinone ring system linked through a methyl\u00adene bridge to a 1,2,3-triazole ring, which in turn carries an n-butyl substituent. The triazole ring is inclined by 67.09\u2005(4)\u00b0 to the quinoxalinone ring plane. In the crystal, the mol\u00adecules form oblique stacks along the a-axis direction through inter\u00admolecular C\u2014HTrz\u22efNTrz (Trz = triazole) hydrogen bonds, and offset \u03c0-stacking inter\u00adactions between quinoxalinone rings [centroid\u2013centroid distance = 3.9107\u2005(9)\u2005\u00c5] and \u03c0\u2013\u03c0 inter\u00adactions, which are associated pairwise by inversion-related C\u2014HDhydqn\u22ef\u03c0(ring) inter\u00adactions. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (52.7%), H\u22efN/N\u22efH (18.9%) and H\u22efC/C\u22efH (17.0%) inter\u00adactions.The title compound, C Diversely substituted quinoxalines and their derivatives embedded with variety of functional groups are important biological agents and a significant amount of research activity has been directed towards this class of compounds. These mol\u00adecules exhibit a wide range of biological applications and are potentially useful in medicinal chemistry research and have therapeutic applications such as anti\u00admicrobial methyl substituent projecting well out of the mean plane of the di\u00adhydro\u00adquinxalone unit, as indicated by the C1\u2014N2\u2014C10\u2014C11 torsion angle of 90.85\u2005(16)\u00b0. The n-butyl group is oriented in the opposite direction as seen from the N4\u2014N3\u2014C13\u2014C14 torsion angle of \u221295.26\u2005(16)\u00b0 N3 Fig.\u00a01. The di\u00ad)\u00b0 Fig.\u00a02.a-axis direction through inter\u00admolecular C\u2014HTrz\u22efNTrz (Trz = triazole) hydrogen bonds (Table\u00a01A (C1\u2013C6) and B (N1/N2/C1/C6\u2013C8) rings and \u03c0-inter\u00adactions between the C8=O1 carbonyl group and the B rings . Pairs of stacks are associated through C\u2014HDhydqn\u22ef\u03c0 inter\u00adactions, generating small, diamond-shaped channels along the a-axis direction generated 37 hits. Of these, the ones most comparable to the title mol\u00adecule have R1 = CH3 and R = CH2C\u2261CH methyl methyl isomer shows that the latter has a U shape with the R group extending back over the bicyclic unit as the result of an intra\u00admolecular C\u2014H\u22efO hydrogen bond from the \u03b1 hydrogen of the butyl group while in the former, the more remote position of the butyl group on the triazole ring disfavours such an inter\u00adaction and the mol\u00adecule adopts a Z shape. This conformation is favoured by the opportunity for \u03c0-stacking and C\u2014H\u22ef\u03c0(ring) inter\u00adactions in the crystal.A search of the CSD , and those delineated into H\u22efH, H\u22efN/N\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH, C\u22efC, O\u22efC/C\u22efO, N\u22efC/C\u22efN and N\u22efN contacts \u2013(i), respectively, together with their relative contributions to the Hirshfeld surface. The most important inter\u00adaction is H\u22efH contributing 52.7% to the overall crystal packing, which is reflected in Fig.\u00a06b) as widely scattered points of high density due to the large hydrogen content of the mol\u00adecule. The split spike with the tip at de = di = 1.13\u2005\u00c5 in Fig.\u00a06b) is due to the short inter\u00adatomic H\u22efH contacts (Table\u00a02c), contribute 18.9% to the HS (Table\u00a02de + di = 2.23\u2005\u00c5. In the presence of weak C\u2014H\u22ef\u03c0 inter\u00adactions (Table\u00a01d), with the tips at de + di = 2.65\u2005\u00c5 (Table\u00a02e)] contacts analysis \u2013(d), respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efN/N\u22efH, H\u22efC/C\u22efH and H\u22efO/O\u22efH inter\u00adactions suggests that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing (Hathwar H)-one (0.68\u2005mmol) in ethanol (15\u2005mL) was added 1-azido\u00adbutane (1.03\u2005mmol). The reaction mixture was stirred under reflux for 72\u2005h. After completion of the reaction (monitored by TLC), the solution was concentrated and the residue was purified by column chromatography on silica gel by using as eluent the mixture (hexa\u00adne/ethyl acetate 8:2). The solid product obtained was crystallized from ethanol to afford colourless crystals in 78% yield.To a solution of 3-methyl-1-(prop-2-yn\u00adyl)-3,4-di\u00adhydro\u00adquinoxalin-2 I, global. DOI: 10.1107/S205698901801589X/xu5949Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901801589X/xu5949Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S205698901801589X/xu5949Isup4.cmlSupporting information file. DOI: 1878133CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Parallel chains inter\u00adact through N\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking of the tris\u00adubstituted phenyl rings.In the title pyrazoline derivative, the pyrazoline ring makes angles of 86.73\u2005(12) and 13.44\u2005(12)\u00b0 with the tris\u00adubstituted and disubstituted benzene rings, respectively. In the crystal structure, the mol\u00adecules are connected into chains running in the 16H16N2O3\u00b7H2O, the pyrazoline ring has an envelope conformation with the substituted sp2 C atom on the flap. The pyrazoline ring makes angles of 86.73\u2005(12) and 13.44\u2005(12)\u00b0 with the tris\u00adubstituted and disubstituted benzene rings, respectively. In the crystal structure, the mol\u00adecules are connected into chains running in the b-axis direction by O\u2014H\u22efN hydrogen bonding. Parallel chains inter\u00adact through N\u2014H\u22efO hydrogen bonds and \u03c0\u2013\u03c0 stacking of the tris\u00adubstituted phenyl rings. The major contribution to the surface contacts are H\u22efH contacts (44.3%) as concluded from a Hirshfeld surface analysis.In the title pyrazoline derivative, C There is a clear difference in both C\u2014N bond distances in the pyrazoline ring: N1=C5 shows double-bond character [1.287\u2005(3)\u2005\u00c5] while N2\u2014C3 [1.496\u2005(3)\u2005\u00c5] is a single bond. The dihedral angle between the two benzene rings is 80.66\u2005(11)\u00b0. The planes of the C6\u2013C11 benzene ring (r.m.s. deviation = 0.004\u2005\u00c5) and the pyrazoline ring make an angle of 86.73\u2005(12)\u00b0. For the C15\u2013C20 benzene ring (r.m.s. deviation = 0.006\u2005\u00c5), the dihedral angle with the pyrazoline ring is only 13.44\u2005(12)\u00b0. Both the hy\u00addroxy and meth\u00adoxy substituents of the C6\u2013C11 phenyl group are within the phenyl plane with deviations of 0.011\u2005(1) (O12), 0.166\u2005(2) (C13) and \u22120.057\u2005(2)\u2005\u00c5 (O14).The title compound crystallizes in the ortho\u00adrhom\u00adbic space group it Fig.\u00a01. The pyr2, the O22 water mol\u00adecule bridges three mol\u00adecules by O\u2014H\u22efN and O\u2014H\u22efO hydrogen-bonding inter\u00adactions with the N1 atom and the O21-hy\u00addroxy group = 3.6627\u2005(11)\u2005\u00c5; slippage 1.442\u2005\u00c5; Cg1 is the centroid of the C6\u2013C11 ring]. In addition, a C\u2014H\u22efO inter\u00adaction is observed in the crystal packing . The associated two-dimensional fingerprint plots followed by recip\u00adrocal C\u22efH/H\u22efC contacts . Significant contributions come from reciprocal O\u22efH/H\u22efO (20.7%) and N\u22efH/H\u22efN (7.0%) contacts, which appear as two symmetrical spikes at de + di = 1.65 and 1.80\u2005\u00c5, respectively . A further small contribution is from C\u22efC contacts .The Hirshfeld surface -4,5-di\u00adhydro-1H-pyrazol-5-yl]phenol \u00b0, that between the pyrazoline ring and the nitro\u00adphenyl ring is 9.7\u2005(1)\u00b0 and that between the pyrazoline ring and the meth\u00adoxy\u00adphenol ring is 56. 78\u2005(9)\u00b0. The second structure, 3-(2\u2032-hy\u00addroxy-5\u2032-meth\u00adoxy-phen\u00adyl)-5-(3-meth\u00adoxy-4-hy\u00addroxy\u00adphen\u00adyl)-4,5-di\u00adhydro-1H-pyrazole \u00b0, while the dihedral angles between the pyrazoline ring and the phenyl rings at atom C3 and C5 are 12.1\u2005(1) and 68.2\u2005(1)\u00b0, respectively.A search of the Cambridge Structural Database -3-(4-Hy\u00addroxy-3-meth\u00adoxy\u00adphen\u00adyl)-1-(4-hy\u00addroxy\u00adphen\u00adyl)prop-2-en-1-one, 1, was synthesized as described in a previous study -4,5-di\u00adhydro-1H-pyrazol-5-yl)-2-meth\u00adoxy\u00adphenol (2):1 (0.01\u2005mol), 2.5\u2005mL of hydrazine hydrate and 25\u2005mL of ethanol was refluxed at 353\u2005K for 2\u2005h. After pouring the reaction mixture into 200\u2005mL of ice\u2013water, the crude solid product was isolated by vacuum filtration, washed several times with cold water and recrystallized from ethanol:water (1:2) to give yellow crystals , m.p. 465\u2005K. 1H NMR [Bruker XL-500, 500\u2005MHz, d6-DMSO, \u03b4 (ppm), J (Hz), see Fig.\u00a06d, 1H, J = 8.0, H2); 3.75 ; 6.74 ; 6.95 , 4.67 ; 3.30 ; 2.76 , 7.45 ; 6.76 .A mixture of chalcone A and H22B were found in difference electron density maps and refined freely. The other H atoms were placed in idealized positions and included as riding contributions with Uiso(H) values of 1.2Ueq or 1.5Ueq of the parent atoms, with C\u2014H distances of 0.93 (aromatic), 0.98 (CH), 0.97 (CH2) and 0.96\u2005\u00c5 (CH3). In the final cycles of refinement, eight outliers were omitted.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019013379/sj5579sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019013379/sj5579Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019013379/sj5579Isup3.cmlSupporting information file. DOI: 1956698, 1956698CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "S(6) ring motif. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds, forming layers parallel to (011). \u03c0\u2013\u03c0 stacking inter\u00adactions complete the three-dimensional network.The asymmetric unit contains two crystallographically independent mol\u00adecules in which the dihedral angles between the benzene rings are 13.26\u2005(5) and 7.87\u2005(5)\u00b0. An intra\u00admolecular O\u2014H\u22efN hydrogen bonds results in the formation of an 15H12N2O2, contains two crystallographically independent mol\u00adecules in which the dihedral angles between the benzene rings in each are 13.26\u2005(5) and 7.87\u2005(5)\u00b0. An intra\u00admolecular O\u2014H\u22efN hydrogen bonds results in the formation of an S(6) ring motif. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds, forming layers parallel to (011). In addition, \u03c0\u2013\u03c0 stacking inter\u00adactions with centroid\u2013centroid distances in the range 3.693\u2005(2)\u20133.931\u2005(2)\u2005\u00c5 complete the three-dimensional network.The asymmetric unit of the title compound, C The title compound displays a trans configuration with respect to the C8=N1 and C23=N3 double bonds. In each independent mol\u00adecule, an intra\u00admolecular O\u2014H\u22efN hydrogen bond (Table\u00a01G (N1/H2/O2/C2/C7/C8) and H (O4/H4/N3/C23/C22/C17)]; these are oriented at dihedral angles of A/G = 1.31\u2005(5) and C/H = 0.42\u2005(5)\u00b0 with respect to the adjacent benzene rings.The asymmetric unit of the title compound contains two crystallographically independent mol\u00adecules = 3.860\u2005(2)\u2005\u00c5, Cg2\u22efCg2 = 3.693\u2005(2)\u2005\u00c5 and Cg2\u22efCg4 = 3.931\u2005(2)\u2005\u00c5; where Cg1, Cg2, Cg3 and Cg4 are the centroids of the C2\u2013C7, C9\u2013C14, C17\u2013C22 and C24\u2013C29 rings, respectively Fig.\u00a02. In addily Fig.\u00a03.\u03c0\u2013\u03c0* type transition. The frontier orbital gap helps to characterize the chemical reactivity and the kinetic stability of the mol\u00adecule. A mol\u00adecule with a small frontier orbital gap is generally associated with a high chemical reactivity, low kinetic stability and is also termed a soft mol\u00adecule. DFT quantum-chemical calculations for the title compound were performed at the B3LYP/6\u2013311G level and the lowest lying unoccupied mol\u00adecular orbitals (LUMOs) are termed frontier mol\u00adecular orbitals (FMOs), which play an important role in the optical and electric properties of compounds, as well as in their quantum chemistry and UV\u2013vis spectra. According to mol\u00adecular orbital theory, an inter\u00adaction between HOMO and LUMO orbitals of a structure gives rise to a et al., 2002Mol\u00adecular electrostatic potential (MEP) surface analysis is a technique of mapping electrostatic potential onto the iso-electron density surface, providing information about the reactive sites. The surface simultaneously displays mol\u00adecular size and shape and the electrostatic potential value. In the colour scheme adopted, red indicates an electron-rich region with a partially negative charge and blue an electron-deficient region with partially positive charge, light blue indicates a slightly electron-deficient region, yellow a slightly electron-rich region and green a neutral region -2-[(2-hy\u00addroxy-5-meth\u00adoxy\u00adbenzyl\u00adidene)amino]\u00adbenzo\u00adnitrile moiety: (Z)-2-[(2-hy\u00addroxy-1-naphth\u00adyl)methyl\u00adene\u00adamino]\u00adbenzo\u00adnitrile -2-[(5-bromo-2-hy\u00addroxy\u00adbenzyl\u00adidene)amino]\u00adbenzo\u00adnitrile \u00adbenzo\u00adnitrile\u00ad-2-\u00adbenzo\u00adnitrile -2-(4-di\u00adethyl\u00adamino-2-hy\u00addroxy\u00adbenzyl\u00adidene\u00adamino)\u00adbenzo\u00adnitrile -2-[amino]\u00adbenzo\u00adnitrile ring motif, similar to title compound.A search of the Cambridge Structural Database in ethanol (15\u2005ml) and 2-amino\u00adbenzo\u00adnitrile in ethanol (15\u2005ml). The reaction mixture was stirred for 5\u2005h under reflux. Single crystals of the title compound suitable for X-ray analysis were obtained by slow evaporation of an ethanol solution .Uiso(H) = 1.2Ueq(C) or 1.5Ueq.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019008077/lh5907sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019008077/lh5907Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019008077/lh5907Isup3.cmlSupporting information file. DOI: 1912294CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The ribbons are connected into layers parallel to the bc plane by inversion-related C\u2014HBnz\u22ef\u03c0(ring) inter\u00adactions.The pyridazine ring deviates slightly from planarity. In the crystal, ribbons consisting of inversion-related chains of mol\u00adecules extending along the 22H16N4O2, contains two pyridine rings and one meth\u00adoxy\u00adcarbonyl\u00adphenyl group attached to a pyridazine ring which deviates very slightly from planarity. In the crystal, ribbons consisting of inversion-related chains of mol\u00adecules extending along the a-axis direction are formed by C\u2014HMthy\u22efOCarbx (Mthy = methyl and Carbx = carboxyl\u00adate) hydrogen bonds. The ribbons are connected into layers parallel to the bc plane by C\u2014HBnz\u22ef\u03c0(ring) (Bnz = benzene) inter\u00adactions. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (39.7%), H\u22efC/C\u22efH (27.5%), H\u22efN/N\u22efH (15.5%) and O\u22efH/H\u22efO (11.1%) inter\u00adactions. Hydrogen-bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Computational chemistry indicates that in the crystal, C\u2014HMthy\u22efOCarbx hydrogen-bond energies are 62.0 and 34.3\u2005kJ\u2005mol\u22121, respectively. Density functional theory (DFT) optimized structures at the B3LYP/6-311G level are com\u00adpared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.The title com\u00adpound, C O-triphosphate can be used as a potential substrate for fluorescence detection and imaging of DNA pyridazine derivatives are a versatile class of nitro\u00adgen-containing heterocyclic com\u00adpounds and they constitute useful inter\u00admediates in organic syntheses. Also, this nucleus is one of the important ligands in the field of coordination chemistry research. 5--2\u2032-de\u00adoxy\u00aduridine-5\u2032-B (atoms N2/N3/C6\u2013C9), deviates slightly from planarity by \u00b10.021\u2005(1)\u2005\u00c5 (r.m.s. deviation = 0.0134\u2005\u00c5) and C (N4/C10\u2013C14), are inclined to the mean plane of the pyridazine ring, B, by 18.68\u2005(6) and 38.40\u2005(6)\u00b0, respectively, while the benzene ring, D (C15\u2013C20), is inclined to it by 62.59\u2005(5)\u00b0. The pyridine and benzene rings are oriented at dihedral angles of A/C = 25.16\u2005(4)\u00b0, A/D = 48.94\u2005(4)\u00b0 and C/D = 59.13\u2005(4)\u00b0. The plane of the carboxyl group (defined by atoms C18/C21/O1/O2) is twisted out of the plane of the benzene ring, D, by 22.88\u2005(5)\u00b0.The title com\u00adpund contains two pyridine rings and one meth\u00adoxy\u00adcarbonyl\u00adphenyl group attached to a pyridazine ring, where the central pyridazine ring, \u00c5) Fig.\u00a01. The plaa-axis direction are formed by C22\u2014H22C\u22efO1v hydrogen bonds and those delineated into H\u22efH, H\u22efC/C\u22efH, H\u22efN/N\u22efH, H\u22efO/O\u22efH, C\u22efC and C\u22efN/N\u22efC contacts \u2013(g), respectively, together with their relative contributions to the Hirshfeld surface. The most important inter\u00adaction is H\u22efH contributing 39.7% to the overall crystal packing, which is reflected in Fig.\u00a06b) as widely scattered points of high density due to the large hydrogen content of the mol\u00adecule with the tip at de = di = 1.10\u2005\u00c5, due to the short inter\u00adatomic H\u22efH contacts (Table\u00a02c) with the tips at de + di = 2.75\u2005\u00c5. The pair of scattered points of wings resulting in the fingerprint plots delineated into H\u22efN/N\u22efH contacts, with a 15.5% contribution to the HS, has a symmetrical distribution of points with the edges at de + di = 2.58\u2005\u00c5 (Table\u00a02e), with an 11.1% contribution to the HS, arises from the O\u22efH/H\u22efO contacts have an arrow-shaped distribution of points with the tip at de = di = 13.50\u2005\u00c5. Finally, the tiny characteristic wings resulting in the fingerprint plots shown in Fig.\u00a06g, a 2.4% contribution to the HS, arises from the C\u22efN/N\u22efC contacts analysis \u2013(d), respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH, H\u22efN/N\u22efH and H\u22efO/O\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing is the sum of the electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange\u2013repulsion (Erep) energies were calculated as \u221223.9 (Eele), \u22124.3 (Epol), \u221276.2 (Edis), 53.0 (Erep) and \u221262.0 (Etot) for the C22\u2014H22C\u22efO1 hydrogen-bonding inter\u00adaction, and \u221222.0 (Eele), \u22128.5 (Epol), \u221228.5 (Edis), 32.3 (Erep) and \u221234.3 (Etot) for the C22\u2014H22B\u22efO1 hydrogen-bonding inter\u00adaction.The inter\u00admolecular inter\u00adaction energies were calculated using the CE\u2013B3LYP/6-31G energy model available in via density functional theory (DFT) using the standard B3LYP functional and 6-311G basis-set calculations , hardness (\u03b7), potential (\u03bc), electrophilicity (\u03c9) and softness (\u03c3) are all recorded in Table\u00a04E = ELUMO \u2212 EHOMO] of the mol\u00adecule is about 1.8908\u2005eV, and the frontier mol\u00adecular orbital (FMO) energies, i.e. EHOMO and ELUMO, are \u22124.3680 and \u22122.4772\u2005eV, respectively.The optimized structure of the title com\u00adpound, (I)et al., 2019a2N1,N6]copper(II) bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfonate) pyridazine]bis\u00ad(\u03bc-2-azido)\u00addizaidodicopper monohydrate] meth\u00adyl]phenyl analogue has been reported to afford colourless crystals .3,6-Bis(pyridin-2-yl)-1,2,4,5-tetra\u00adzine (4\u2005mmol) was dissolved in toluene (20\u2005ml), and then 1 equiv. of methyl 4-ethynylbenzoate was added and the reaction mixture was stirred and refluxed at temperatures between 413 and 453\u2005K. The solvent was then evaporated. The product obtained was separated by chromatography on a column of silica gel. The isolated solid was recrystallized from hexa\u00adne\u2013di\u00adchloro\u00admethane I, global. DOI: 10.1107/S2056989019013732/lh5927Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019013732/lh5927Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019013732/lh5927Isup4.cmlSupporting information file. DOI: 1958277, 1958277CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The pyrrolidine ring makes a dihedral angle of 14.83\u2005(12)\u00b0 with the 3,4-di\u00admeth\u00adoxy\u00adphenyl ring, which are attached to each other by an extended N\u2014CH2\u2014CH2\u2014Car bridge. In the crystal, the structure features C\u2014H\u22efO inter\u00admolecular hydrogen bonds, an offset \u03c0\u2013\u03c0 inter\u00adaction [inter\u00adcentroid distance = 3.564\u2005(1)\u2005\u00c5] and a C\u2014Cl\u22ef\u03c0 inter\u00adaction. The contribution of some disordered solvent to the scattering was removed using the SQUEEZE routine [Spek (2015PLATON. The solvent contribution was not included in the reported mol\u00adecular weight and density.In the title compound, CSpek 2015. Acta Cr The two five-membered rings, A (C2/C3/C5\u2013C7) and B (C5\u2013C9), have envelope conformations with atom C6 as the flap: puckering parameters and the smallest displacement asymmetric parameters are Q2 = 0.619\u2005(2)\u2005\u00c5, \u03c62 = 108.6\u2005(2)\u00b0 and \u0394s = 1.09\u00b0 for ring A, and Q2 = 0.582\u2005(2)\u2005\u00c5, \u03c62 = 215.5\u2005(2)\u00b0 and \u0394s = 0.74\u00b0 for ring B. Atom C6 is displaced from the mean plane through the other four atoms by 0.908\u2005(2)\u2005\u00c5 in ring A and 0.875\u2005(2)\u2005\u00c5 in ring B. The dihedral angle between the pyrrolidine ring (N1/C1\u2013C4) and the benzene ring (C12\u2013C17) is 14.83\u2005(12)\u00b0, with the torsion angle N1\u2014C10\u2014C11\u2014C12 being 175.8\u2005(3)\u00b0. The lengths of the C\u2014Cl bonds involving the chlorine atoms attached to the C8=C9 double bond are 1.692\u2005(2)\u2005\u00c5 for C8\u2014Cl2 and 1.692\u2005(2)\u2005\u00c5 for C9\u2014Cl3. The lengths of the bonds to chlorine atoms attached to the single C\u2014C bonds vary from 1.744\u2005(2) to 1.768\u2005(2)\u2005\u00c5. These value are close to those found in similar compounds; see \u00a74 Database survey.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01A\u22efO2i hydrogen bonds link the molecules to form a cyclic Cg1\u22efCg5iii of 3.564\u2005(1)\u2005\u00c5 dec-8-ene-3,5-dione \u00b0 compared to 175.8\u2005(3)\u00b0 in the title compound.A search of the Cambridge Structural Database \u20131.692\u2005(2)\u2005\u00c5 and 1.744\u2005(2)\u20131.768\u2005(2)\u2005\u00c5, respectively.In all 17 structures, the five-membered ring has envelope conformations and the six-membered ring a boat conformation. The bond lengths and bond angles are very similar to those reported here for the title compound. For example, the C2CO3 and dried over anhydrous Na2SO4. It was then filtered and the filtrate was concentrated under reduced pressure followed by silica gel column purification to afford the title compound in 82% yield. Colourless block-shaped crystals were obtained by slow evaporation of a solution in ethanol.2- ethanamine (1 equiv.) and 1,4,5,6,7,7-hexa\u00adchloro-5- norbornene \u22122,3-di\u00adcarb\u00adoxy\u00adlic anhydride (1 equiv.) were stirred at room temperature in dry ethyl acetate for 30\u2005min. The ethyl acetate was removed under reduced pressure and the resulting residue was dissolved in toluene. To this reaction mixture was added acetyl chloride (5 equiv.) and refluxed for 1\u2005h. The reaction mixture was brought to room temperature and washed with aqueous NaUiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. The contribution of the disordered solvent to the scattering was removed using the SQUEEZE routine of PLATON Global, I. DOI: 10.1107/S2056989019004109/su5485Isup3.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019004109/su5485sup4.pdfCSD search S1. DOI: Click here for additional data file.10.1107/S2056989019004109/su5485Isup4.cmlSupporting information file. DOI: 1905872CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, C\u2014HOxqn\u22efOEthx and C\u2014HPh\u00adyl\u22efOCarbx weak hydrogen bonds link the mol\u00adecules into a three-dimensional network sturucture. A \u03c0\u2013\u03c0 inter\u00adaction between the constituent rings of the oxo\u00adquinoline unit, with a centroid\u2013centroid distance of 3.675\u2005(1)\u2005\u00c5 may further stabilize the structure. Both terminal ethyl groups are disordered over two sets of sites. The ratios of the refined occupanies are 0.821\u2005(8):0.179\u2005(8) and 0.651\u2005(18):0.349\u2005(18). The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (53.9%), H\u22efO/O\u22efH (28.5%) and H\u22efC/C\u22efH (11.8%) inter\u00adactions. Weak inter\u00admolecular hydrogen-bond inter\u00adactions and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Density functional theory (DFT) geometric optimized structures at the B3LYP/6-311G level are com\u00adpared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO mol\u00adecular orbital behaviour was elucidated to determine the energy gap.The title com\u00adpound, C Quinolone derivatives have constituted an important class of heterocyclic com\u00adpounds which, even when part of a com\u00adplex mol\u00adecule, possesses a wide spectrum of biological activities, such as anti\u00adcancer and B (C4\u2013C9), of the oxo\u00adquinoline unit are oriented at a dihedral angle of 1.04\u2005(6)\u00b0. Thus, they are almost coplanar, with a maximum deviation of 0.017\u2005(3)\u2005\u00c5 for atom C7. Atoms O1 and C10 deviate only by 0.007\u2005(2) and 0.022\u2005(2)\u2005\u00c5 from that plane and so are essential coplanar. The acetate substituent is nearly perpendicular to that plane, with a torsion angle of C1\u2014N1\u2014C10\u2014C11 = \u2212104.8\u2005(2)\u00b0. The mean plane of the phenyl ring, C (C19\u2013C24), is oriented with respect to the oxo\u00adquinoline unit at a dihedral angle of 68.17\u2005(6)\u00b0. The carboxyl groups, O5/O6/C11 and O3/O4/C16, are twisted out of coplanarity with the best least-squares plane of the oxo\u00adquinoline unit and phenyl ring C by dihedral angles of 79.7\u2005(2) and 62.9\u2005(2)\u00b0, respectively.The title mol\u00adecule is com\u00adposed of ethyl 2-acetate and 4-[(2-eth\u00adoxy-2-oxoeth\u00adyl)(phen\u00adyl)carbomo\u00adyl] units Fig.\u00a01. The meaOxqn\u22efOEthx and C\u2014HPh\u00adyl\u22efOCarbx hydrogen bonds (Table\u00a01i.e.A (N1/C1\u2013C4/C9) and B (C4\u2013C9), of the oxo\u00adquinoline unit, with Cg1\u22efCg2i = 3.675\u2005(1)\u2005\u00c5 , may further stabilize the structure. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for crystal packing are from H\u22efH (53.9%), H\u22efO/O\u22efH (28.5%) and H\u22efC/C\u22efH (11.8%) inter\u00adactions. Weak inter\u00admolecular hydrogen-bond inter\u00adactions and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing.In the crystal, weak C\u2014Hs Table\u00a01 link thes Table\u00a01. A \u03c0\u2013\u03c0 cCrystalExplorer17.5 and those delineated into H\u22efH, H\u22efO/O\u22efH, H\u22efC/C\u22efH, C\u22efC and O\u22efC/C\u22efO contacts \u2013(f), respectively, together with their relative contributions to the Hirshfeld surface. The most important inter\u00adaction is H\u22efH, contributing 53.9% to the overall crystal packing, which is reflected in Fig.\u00a06b) as widely scattered points of high density due to the large hydrogen content of the mol\u00adecule, with the tip at de = di = 1.05\u2005\u00c5, due to the short inter\u00adatomic H\u22efH contacts. The pair of characteristic wings resulting in the fingerprint plot delineated into H\u22efO/O\u22efH contacts has a 28.5% contribution to the HS and is viewed as a pair of spikes with the tips at de + di = 2.30\u2005\u00c5. In the absence of weak C\u2014H\u22ef\u03c0 inter\u00adactions, the pair of characteristic wings resulting in the fingerprint plot delineated into H\u22efC/C\u22efH contacts , with a 11.8% contribution to the HS and are viewed as a pair of spikes with the tip at de + di = 2.83\u2005\u00c5. The C\u22efC contacts have an arrow-shaped distribution of points with the tip at de = di = 1.81\u2005\u00c5. Finally, the pair of the scattered points of wings from the fingerprint plot are delineated into O\u22efC/C\u22efO contacts, with a 1.1% contribution to the HS, and has a nearly symmetrical distribution of points with the edges at de + di = 3.15\u2005\u00c5.In order to visualize the inter\u00admolecular inter\u00adactions in the title com\u00adpound, a Hirshfeld surface (HS) analysis \u2013(d), respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of weak H-atom contacts in establishing the packing structure. The large number of H\u22efH, H\u22efO/O\u22efH and H\u22efC/C\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and weak hydrogen-bond inter\u00admolecular inter\u00adactions play major roles in the crystal packing com\u00adputational calculations using a standard B3LYP functional and a 6-311G basis set , hardness (\u03b7), potential (\u03bc), electrophilicity (\u03c9) and softness (\u03c3), which are listed in Table\u00a03E = ELUMO \u2212 EHOMO] of the mol\u00adecule was about 4.2091\u2005eV, and the frontier mol\u00adecular orbital (FMO) energies, i.e. EHOMO and ELUMO, were \u22126.1141 and \u22121.9050\u2005eV, respectively.The geometry optimized structure of the title com\u00adpound in the gas phase was generated theoretically et al., 2016bet al., 2019aet al., 2014et al., 2012et al., 2018aet al., 2015A non-a\u00adlkylated analogue, namely quinoline and its derivatives, has been reported in di\u00admethyl\u00adformamide were added ethyl bromo\u00adacetate (4.16\u2005mmol), K2CO3 (5.67\u2005mmol) and tetra\u00adbutyl\u00adammonium bromide . The reaction mixture was stirred at room temperature for 6\u2005h. After removal of the salts by filtration, the DMF was evaporated under reduced pressure and the resulting residue was dissolved in di\u00adchloro\u00admethane. The organic phase was dried with Na2SO4 and then concentrated under reduced pressure. The pure com\u00adpound was obtained by column chromatography using as eluate hexa\u00adne/ethyl acetate (3:1 v/v). The isolated solid was recrystallized from hexa\u00adne\u2013diethyl acetate (1:1 v/v) to afford colourless crystals .To a solution of 2-oxo-2 and CH3 H atoms, respectively, and constrained to ride on their parent atoms, with Uiso(H) = kUeq(C), where k = 1.5 for CH3 H atoms and k = 1.2 for other H atoms. The terminal ethyl groups are disordered with an occupancy ratio of 0.821\u2005(8):0.179\u2005(8) for C12 and C13, and 0.651\u2005(18):0.349\u2005(18) for C17 and C18.The experimental details, including the crystal data, data collection and refinement, are summarized in Table\u00a0410.1107/S2056989019014154/lh5924sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019014154/lh5924Isup2.hklStructure factors: contains datablock(s) I. DOI: 1959642, 1959642CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Glycyrrhiza on the Diuretic Function of Euphorbia kansui: An Ascites Mouse Model\u201d [Y1 = 6.331\u2217X12\u2217X2 \u2212 4.16\u2217X12 \u2212 0.1637/X22 + 10.94 , displayed in Figure 3(a). Ascites volume/body weight of mice was defined as a dependent variable Y2 using the multivariate stepwise regression equation Y2 = 0.06416\u2217X1\u2217X22 \u2212 0.006046/X22 + 0.3062 as shown in Figure 4(a).\u201d should be corrected toIn the article titled \u201cEffect ofe Model\u201d , there wY1 = 6.331\u2217X12\u2217X2 \u2212 4.16\u2217X12 \u2212 0.1637/X22 + 11.62 , displayed in Figure 3(a). Ascites volume/body weight of mice was defined as a dependent variable Y2 using the multivariate stepwise regression equation Y2 = 0.06416\u2217X1\u2217X22 \u2212 0.006046/X22 + 0.4288 as shown in Figure 4(a).\u201d\u201cAscites volume was defined by the stepwise equation, In addition, the photograph of the renal pathological changes in the Furosemide group in Figure 7(b)-Furo was misplaced. The corrected version of the figure with its description is shown below:"} +{"text": "II atom bridging two six-coordinate ZnII atoms in which the two terminal ZnII cations adopt distorted octa\u00adhedral geometries and the central ZnII cation adopts a distorted tetra\u00adhedral geometry.The structure is reported of a complex containing a trinuclear Zn cation lying on a crystallographic twofold axis. It consists of a tetra\u00adhedral Zn 3(C14H11Br2N2O)4](ClO4)2\u00b72CH3CN, crystallizes as a symmetrical trinuclear cation with all three metal atoms being located on a twofold rotation axis. It contains a tetra\u00adhedral ZnII atom that bridges two six-coordinate ZnII atoms. The complex contains N- and O-donor atoms of four tridentate 2,4-di\u00adbromo-6-{[2-(pyridin-2-yl)eth\u00adyl]imino\u00admeth\u00adyl}phenolate ligands. The ratio of ZnII atoms to ligands is 3:4. The two terminal ZnII cations adopt distorted octa\u00adhedral geometries and the central ZnII cation adopts a distorted tetra\u00adhedral geometry. In the cation there are \u03c0\u2013\u03c0 inter\u00adactions between the di\u00adbromo\u00adphenyl rings, as well as halogen-bonding inter\u00adactions between the di\u00adbromo\u00adphenyl rings in the cation, which stabilize its conformation. In addition, there are C\u2014H\u22efO inter\u00adactions between the anions and both the cations and solvent mol\u00adecules as well as C\u2014H\u22efN inter\u00adactions between the cation and solvent mol\u00adecules. These inter\u00adspecies inter\u00adactions link the cations, anions and solvent mol\u00adecules into a complex three-dimensional arrayThe title compound, [Zn Some enzymes containing zinc(II) include carbonic anhydrase, carb\u00adoxy\u00adpeptidase, and phosphatase 4](ClO4)2\u00b72CH3CN, 1, contains a complex cation as well as perchlorate anions and aceto\u00adnitrile solvent mol\u00adecules and thus has an overall stoichiometry of [Zn3(L)4](ClO4)2.2CH3CN where L is 2,4-di\u00adbromo-6-{[(2-(pyridin-2-yl)eth\u00adyl]imino\u00admeth\u00adyl}phenolate. The compound crystallizes in the monoclinic space group C2/c and the cation consists of the four equivalent L ligands, uniformly coordin\u00adated to three ZnII cations.The crystal structure of the title compound, [Zn3(L)4]2+, lies on a crystallographic twofold axis to 120.11\u2005(8)\u00b0 and Zn\u2014O bond lengths of 1.9512\u2005(19) and 1.9602\u2005(19)\u2005\u00c5. For the six-coordinate terminal zinc atoms, as is usual for complexes containing both Schiff base imine and pyridine N donors, the former form shorter bonds [Zn1\u2014N1 = 2.122\u2005(2)\u2005\u00c5 and Zn3\u2014-N3 = 2.067\u2005(2)\u2005\u00c5] while the latter form longer bonds [Zn1\u2014N2 = 2.148\u2005(2)\u2005\u00c5 and Zn3\u2014N4 = 2.177\u2005(2)\u2005\u00c5] to zinc. The metrical parameters involving the bridging phenolate O donors are significantly different. The bonds to the central Zn2 are considerably shorter than those to the terminal Zn1 and Zn3 and the bridging angles are Zn1\u2014O1\u2014Zn2 = 96.78\u2005(8)\u00b0 and Zn2\u2014O2\u2014Zn3 = 93.73\u2005(8)\u00b0. The distortion from an octa\u00adhedral geometry can be seen from the cis and trans angles which range from 77.49\u2005(10) to 98.19\u2005(9)\u00b0 and 160.47\u2005(13) to 173.41\u2005(12)\u00b0, respectively. Since all three Zn atoms lie on the twofold axis, the Zn1\u2014Zn2\u2014Zn3 bond angle is exactly 180\u00b0. These metrical parameters are similar to those found in the most closely similar complex to 2.117\u2005(4)\u2005\u00c5 and 2.140\u2005(4) to 2.176\u2005(4)\u2005\u00c5, respectively. In this complex there is no crystallographically imposed symmetry; however, the Zn\u2014Zn\u2014Zn bond angle is still close to 180 at 172.51\u2005(3)\u00b0.The trinuclear complex cation, as well as halogen-bonding inter\u00adactions between the di\u00adbromo\u00adphenyl rings in the cation, which stabilize its conformation. In addition there C\u2013H\u22efO inter\u00adactions between the anions and both the cations and solvent mol\u00adecules as well as C\u2014H\u22efN inter\u00adactions between the cation and solvent mol\u00adecules (Table\u00a01In the cation there are \u03c0\u2013\u03c0 inter\u00adactions between the di\u00adbromo\u00adphenyl rings imino}\u00admeth\u00adyl)phenolato type ligands gave 26 hits of which only one was similar to the title compound in that it contained a trinuclear Zn complex where this ligand was acting as a bridging group to the central Zn atom 2\u00b76H2O with no added base. The mixture was stirred at room temperature overnight. The methanol was removed by rotary evaporation. The product was crystallized by slow evaporation of a solution in acetonitrile giving pale-yellow to colorless crystals.2-(2-Pyrid\u00adyl)ethyl\u00adamine was dissolved in 50\u2005mL of methanol. 3,5-Di\u00adbromo\u00adsalicyl\u00adaldehyde was added to the solution and the mixture was refluxed for 5\u2005h. The zinc(II) complex was prepared by reacting the ligand in 50\u2005ml of methanol with Zn(ClOUiso(H) = 1.2Ueq(C) or 1.5Ueq(CH3).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018012100/jj2202sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018012100/jj2202Isup2.hklStructure factors: contains datablock(s) I. DOI: 1863971CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Reaction of this complex with piperidine leads to partial decoordination of the 2-pyridyl\u00adphosphine in the product, [P-{(C6H5)2(C5H5N)P}(C5H11N)Re(CO)3Br], which displays an intra\u00admolecular hydrogen bond between the piperidine aminic hydrogen atom and the uncoordinated pyridyl group, with D\u22efA = 2.992\u2005(9)\u2005\u00c5.The reaction of the ligand di\u00adpyridyl\u00adphosphine with (Re(CO) 2N,P]rhenium(I) chloro\u00adform disolvate, [ReBr(C17H14NP)(CO)3]\u00b72CHCl3 or \u00b72CHCl3, (I\u00b72CHCl3), is best described as a distorted octa\u00adhedron with three carbonyls in a facial conformation, a bromide atom, and a biting P,N-di\u00adphenyl\u00adpyridyl\u00adphosphine ligand. Hirshfeld surface analysis shows that C\u2014Cl\u22efH inter\u00adactions contribute 26%, the distance of these inter\u00adactions are between 2.895 and 3.213\u2005\u00c5. The reaction between I and piperidine (C5H11N) at 313\u2005K in di\u00adchloro\u00admethane leads to the partial decoord\u00adination of the pyridyl\u00adphosphine ligand, whose pyridyl group is replaced by a piperidine mol\u00adecule, and the complex bromido\u00adtricarbon\u00adyl[diphen\u00adyl(pyridin-2-yl)phosphane-\u03baP](piperidine-\u03baN)rhenium(I), [ReBr(C5H11N)(C17H14NP)(CO)3] or [P-{(C6H5)2(C5H5N)P}(C5H11N)Re(CO)3Br] (II). The mol\u00adecule has an intra\u00admolecular N\u2014H\u22efN hydrogen bond between the non-coordinated pyridyl nitro\u00adgen atom and the amine hydrogen atom from piperidine with D\u22efA = 2.992\u2005(9)\u2005\u00c5. Thermogravimetry shows that I\u00b72CHCl3 losses 28% of its mass in a narrow range between 318 and 333\u2005K, which is completely consistent with two solvating chloro\u00adform mol\u00adecules very weakly bonded to I. The remaining I is stable at least to 573\u2005K. In contrast, II seems to lose solvent and piperidine (12% of mass) between 427 and 463\u2005K, while the additional 33% loss from this last temperature to 573\u2005K corresponds to the release of 2-pyridyl\u00adphosphine. The contribution to the scattering from highly disordered solvent mol\u00adecules in II was removed with the SQUEEZE routine [Spek phosphane-\u03baSpek 2015. Acta Cr The mol\u00adecule is a rigid bidentate ligand Br)2 in chloro\u00adform as solvent leads to the complex P,N-{(C6H5)2(C5H5N)P}Re(CO)3Br]\u00b72CHCl3 (I\u00b72CHCl3). It presents a similar structure to the widely studied [Re(CO)3(L)] complexes, which have inter\u00adesting photophysical and photochemical properties , leading to the complex [P-{(C6H5)2(C5H5N)P}(C5H11N)Re(CO)3Br] (II).The reaction of the di\u00adphenyl\u00adpyridyl\u00adphosphine ligand with the rhenium dimer (Re(CO)I complex I with a bidentate P,N (chelating) ligand crystallized from a chloro\u00adform solution in the monoclinic space P21/c. Selected geometrical data are summarized in Table\u00a01I\u00b72CHCl3 is given in Fig.\u00a012Py, a bromide atom in an apical position and three carbonyl carbon atoms in a fac correlation, generating a distorted octa\u00adhedral environment. Additionally, two chloro\u00adform mol\u00adecules crystallize together with the complex mol\u00adecule.The mononuclear ReI complex II, crystallized from a CH2Cl2/CH3CN (2:1) solution in the triclinic space group P2Py ligand, a piperidine C5H11N mol\u00adecule and a bromide anion. The piperidine ring displays a chair-like conformation. An intra\u00admolecular hydrogen bond is defined between the non-coordinated pyridyl nitro\u00adgen atom and the amine hydrogen atom from piperidine, N2\u2014H2N\u22efN1, with D\u22efA = 2.992\u2005(9)\u2005\u00c5 (Table\u00a04B) of the pypridine ring vs 2119.2\u2005(3)\u2005\u00c53 . In this dnorm view , blue represents the longest distances while the shortest distances are depicted as red spots .Compound II Br)2 and (C6H5)2(C5H5N)P were used as provided from supplier (Aldrich), with no purification before use. Seccosolv\u2122 solvents were used without any further purification. Standard Schlenck techniques under argon atmosphere were used for all manipulations.The reagents, (Re(CO)Synthesis of I. 500\u2005mg of (Re(CO)3(OC4H8)Br)2 (0.590\u2005mmol) were dissolved in 5\u2005ml of chloro\u00adform. 312\u2005mg of diphenyl-2-pyridyl\u00adphosphine (1.18\u2005mmol) was dissolved in 10\u2005ml of chloro\u00adform. The two solutions were mixed, changing from colourless to a translucent yellow after 10 minutes of reaction. The reaction was left to continue for a further 2\u2005h. Addition of 2\u2005ml of pentane to the mixture and standing by one day lead to yellow diffraction-quality crystals of I\u00b72CHCl3 .Synthesis of II. The compound was prepared by direct reaction between I and an excess of piperidine (C5H11N) at 313\u2005K in CH2Cl2. 50.0\u2005mg of (0.082\u2005mmol) were dissolved in 10\u2005ml of CH2Cl2 giving rise to a yellow solution. Then, 40\u2005\u00b5L of piperidine (0.51\u2005mmol) was slowly added. The reaction was allowed to continue for six days with constant agitation at 313\u2005K. After cooling, the reaction mixture was layered with aceto\u00adnitrile. Small orange\u2013yellow diffraction-quality crystals were obtained after one week.Uiso(H) = 1.2Ueq(C). For II, the amine hydrogen atom of the piperidine ring was located in a Fourier-difference map and then subsequently refined with a distant constraint of 0.82\u2005\u00c5. During the last stages of the refinement of II, a region of highly disordered electron density was detected within the crystal structure. As no meaningful model could be achieved, SQUEEZE I-2CHCl3, II, Global. DOI: 10.1107/S2056989019008089/nk2249I-2CHCl3sup2.hklStructure factors: contains datablock(s) I-2CHCl3. DOI: 10.1107/S2056989019008089/nk2249IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1921165, 1921164CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Each Cu2(CN)2 unit is connected to six FeII centres via two bridging 2-ethyl\u00adpyrazine mol\u00adecules and four cyanido groups, resulting in the formation of a polymeric three-dimensional bimetallic metal\u2013organic framework, additionally stabilized by Cu\u22efCu contacts.In the title compound, {Fe(Etpz) 6H8N2)2{Cu(CN)2}2]n, the low-spin FeII ion lies at an inversion centre and displays an elongated octa\u00adhedral [FeN6] coordination environment. The axial positions are occupied by two symmetry-related bridging 2-ethyl\u00adpyrazine ligands, while the equatorial positions are occupied by four N atoms of two pairs of symmetry-related cyanide groups. The CuI centre is coordinated by three cyanide carbon atoms and one N atom of a bridging 2-ethyl\u00adpyrazine mol\u00adecule, which form a tetra\u00adhedral coordination environment. Two neighbouring Cu atoms have a short Cu\u22efCu contact [2.4662\u2005(7)\u2005\u00c5] and their coordination tetra\u00adhedra are connected through a common edge between two C atoms of cyanide groups. Each Cu2(CN)2 unit, formed by two neighbouring Cu atoms bridged by two carbons from a pair of \u03bc-CN groups, is connected to six FeII centres via two bridging 2-ethyl\u00adpyrazine mol\u00adecules and four cyanide groups, resulting in the formation of a polymeric three-dimensional metal\u2013organic coordination framework.In the title metal\u2013organic framework, [Fe(C The axial positions are occupied by the N atoms of two symmetry-related 2-ethyl\u00adpyrazine mol\u00adecules [Fe1\u2014N3 = 1.981\u2005(2)\u2005\u00c5]. The low-spin state of the FeII centre at the temperature of experiment (T = 173\u2005K) is confirmed by the Fe\u2014N bond lengths (i.e. < 2.0\u2005\u00c5). Each CuI ion (Cu1ii and Cu1iv) is coord\u00adinated by one bridging 2-ethyl\u00adpyrazine mol\u00adecule via the N atom and by the C atoms of three cyanido groups . The separation between two neighboring Cu atoms is 2.4662\u2005(7)\u2005\u00c5, which is significantly shorter than the sum of the corresponding van der Waals radii 2 pairs, in which two Cu atoms, centred about a twofold rotation axis, are inter\u00adconnected by two \u03bc-CN groups through C atoms. The resulting polymeric three-dimensional metal\u2013organic coordination framework is additionally stabilized by supra\u00admolecular Cu\u22efCu contacts in each Cu2(CN)2 unit.Fig.\u00a03et al., 20162(CN)2 unit, the majority of which are copper monometallic metal\u2013organic frameworks (MOFs). Several bimetallic MOFs are slightly similar to the title compound, namely catena-[bis\u00ad(\u03bc3-chloro)\u00adbis\u00ad(\u03bc3-cyano)\u00adtetra\u00adkis\u00ad(\u03bc2-cy\u00adano)bis\u00ad\u00addicadmium(II)dicopper(I)copper(II)] \u00adtetra\u00adkis\u00ad(\u03bc2-cyano)\u00adtetra\u00adkis\u00ad(di\u00admethyl\u00adformamide\u00adtetra\u00adcopper(I)zinc(II)] \u00adtris\u00ad(\u03bc2-cyano)\u00adbis(\u03bc2-2-ethyl\u00adpyrazine)\u00adtetra\u00adcopper(I)] bridging 2-ethyl\u00adpyrazine mol\u00adecules and (ii) bridging cyano groups, thus forming one-dimensional {Cu(CN)}n chains and double-stranded {Cu(CN)}n ribbons, linked into a network by bridging ethyl\u00adpyrazine ligands.A search through the CSD for 2-ethyl\u00adpyrazine gave 20 hits, in most of which 2-ethyl\u00adpyrazine mol\u00adecule binds to Cu, Ag, Mn or Rh ions. In the majority of compounds containing copper, the 2-ethyl\u00adpyrazine serves as a bridging ligand between two Cu atoms in MOFs. An example closely related to the title structure is 2] in 1\u2005ml of H2O; the second layer was an H2O/EtOH mixture ; the third layer was a solution of Fe(ClO4)2\u00b76H2O and 2-ethyl\u00adpyrazine in 0.5\u2005ml of EtOH. After two weeks, brown crystals were formed in the middle layer. The crystals were kept under the mother solution prior to measurement.Crystals of the title compound were obtained by a slow diffusion within three layers in a 3\u2005ml glass tube. The first layer was a solution of K[Cu(CN)Uiso(H) = 1.2Ueq(C) for aromatic hydrogens, C\u2014H = 0.99\u2005\u00c5 with Uiso(H) = 1.2Ueq(C) for CH2 groups and C\u2014H = 0.98\u2005\u00c5 with Uiso(H) = 1.5Ueq(C) for CH3 groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2056989019009496/rz5262sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019009496/rz5262Isup2.hklStructure factors: contains datablock(s) I. DOI: 1937912CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound is constructed by a 4-hy\u00addroxy\u00adbenzyl ring, a 3,4-di\u00adhydroxy\u00adbenzyl\u00adidenyl ring and a hydrazide-connecting bridge. The overall conformation of the title compound are discussed and compared to the related structures. In the crystal, mol\u00adecules are connected by N\u2014H\u22efO, O\u2014H\u22efO and \u03c0\u2013\u03c0 inter\u00adactions. 14H12N2O4, the azomethine C=N double bond has an E configuration. The hydrazide connecting bridge, (C=O)\u2014(NH)\u2014N=(CH), is nearly planar with C\u2014C\u2014N\u2014N and C\u2014N\u2014N=C torsion angles of \u2212177.33\u2005(10) and \u2212174.98\u2005(12)\u00b0, respectively. The 4-hy\u00addroxy\u00adphenyl and 3,4-di\u00adhydroxy\u00adphenyl rings are slightly twisted, making a dihedral angle of 9.18\u2005(6)\u00b0. In the crystal, mol\u00adecules are connected by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds into a three-dimensional network, while further consolidated via \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.6480\u2005(8) and 3.7607\u2005(8)\u2005\u00c5]. The conformation is compared to those of related benzyl\u00adidene-4-hy\u00addroxy\u00adbenzohydrazide derivatives.In the title benzohydrazide derivative, C The bond lengths and angles of the central hydrazide connecting bridge are consistent with those in related structures \u00b0] and C7\u2014N1\u2014N2\u2014C8 [\u2212174.98\u2005(12)\u00b0] are both in an anti-periplanar conformation, the overall conformation for the hydrazide connecting bridge is almost planar. Furthermore, the 4-hy\u00addroxy\u00adphenyl and 3,4-di\u00adhydroxy\u00adphenyl rings are also coplanar to the corresponding azomethine and carbonyl double bonds, with torsion angles N2\u2014C8\u2014C9\u2014C10 [\u22120.76\u2005(19)\u00b0] and C5\u2014C6\u2014C7\u2014O2 [\u22121.18\u2005(19)\u00b0] both in a syn-periplanar conformation. Those torsion angles result in an overall flat shape of the title compound with the dihedral angle between the terminal benzene rings being 9.18\u2005(6)\u00b0.The title hydrazide derivative, (I), consists of a 4-hy\u00addroxy\u00adphenyl ring, a 3,4-di\u00adhydroxyphenyl ring and a hydrazide (C=O)\u2014(NH)\u2014N=(CH) connecting bridge Fig.\u00a01. The C6\u2014O4\u22efO1iii hydrogen bond and those chains are further connected into two-dimensional plates parallel to the ac plane via N1\u2014H1N1\u22efO3i and O1\u2014H1O1\u22efO2i hydrogen bonds with an aavia an O3\u2014H1O3\u22efO2ii hydrogen bond with an bbCg1\u22efCg2iv = 3.6480\u2005(8)\u2005\u00c5 and Cg1\u22efCg2v = 3.7607\u2005(8)\u2005\u00c5 In the crystal, mol\u00adecules are linked by N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds Table\u00a01 into a tet al., 2016E)-N\u2032-benzenyl\u00adidene-4-hy\u00addroxy\u00adbenzohydrazide as the reference moiety resulted in 31 structures with different substituents at the benzyl\u00adidenyl ring. The different substituent (R) together with selected torsion angles, \u03c41 (C5\u2014C6\u2014C7\u2014O2), \u03c42 (C6\u2014C7\u2014N1\u2014N2), \u03c43 (C7\u2014N1\u2014N2\u2014C8) and \u03c44 (N2\u2014C8\u2014C9\u2014C10) as shown in Fig.\u00a04\u03c42 and \u03c43 are anti-periplanar (151.7\u2013179.8\u00b0), showing that the hydrazide connecting bridges are nearly planar. As for the torsion angle \u03c44, all structures adopt a syn-periplanar conformation (0.6\u201319.6\u00b0). Similar to the title compound, the \u03c41 torsion angles for most of the structures are syn-periplanar (2.0\u201329.1\u00b0). However, there are three outliers whose \u03c41 torsion angles are syn-clinal (34.9\u201350.9\u00b0). By comparing the dihedral angles, the structures can be divided into planar compounds and non-planar compounds . In general, as the hydrazide-connecting bridges are nearly planar, relatively flat \u03c41 and \u03c44 torsion angles are observed in the former compounds, while relatively twisted \u03c41 and \u03c44 torsion angles are observed in the latter.A search of the Cambridge Structural Database max 213, 327\u2005nm; FT\u2013IR (KBr) \u03bd (cm\u22121): 3121 (O\u2014H stretching), 2800 (C\u2014H aromatic stretching), 1615 (amide C=O stretching), 1570 (C=N stretching), 1506 (C=C stretching of aromatic compound) cm\u22121; 1H NMR \u03b4 11.39 , 10.10 , 8.23 , 7.77 , 6.84 , 9.33 , 7.22 , 6.90 , 6.77 .M.p. 572\u2013573\u2005K. UV\u2013Vis (MeOH) \u03bb. C-bound H atoms were positioned geometrically (C\u2014H = 0.93\u2005\u00c5) and refined using a riding model with Uiso(H) = 1.2Ueq(C). All O- and N-bound H atoms were located in a difference-Fourier map and refined freely [O\u2014H = 0.80\u2005(2)\u20130.88\u2005(2)\u2005\u00c5 and N\u2014H = 0.87\u2005(2)\u2005\u00c5].Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019010442/is5518sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019010442/is5518Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019010442/is5518Isup3.cmlSupporting information file. DOI: 1942396CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The best estimator of Eint is the kinetic energy density (Gb), which reasonably approximates the whole set of the structures as \u2212Eint = 0.128Gb2 \u2212 0.82Gb + 1.66 and demonstrates low dispersion. The potential and kinetic energy densities, electron density, and the d(X\u00b7\u00b7\u00b7X) distance behave similarly as estimators of Eint for the individual series Cl\u00b7\u00b7\u00b7Cl, Br\u00b7\u00b7\u00b7Br, and I\u00b7\u00b7\u00b7I. A number of the Eint(property) correlations are recommended for the practical application in the express estimates of the strength of the homo-halogen bonds.Relationships between interaction energy (E Halogen bonds play a very important role in molecular recognition and crystal engineering, synthesis of functional solid materials \u2212 type formed upon interaction of the neutral fragment (A)nZ\u2013Y and the halide anion X\u2212 were considered. It was shown that the Eint(property) correlations are different for each particular type of the Y\u00b7\u00b7\u00b7X\u2212 interaction and they are also different from Equations (1)\u2013(4). Several Eint(property) relationships practically important for express estimates of the interaction energy were recommended for each series of these structures.Recently, the author started the project aimed to establish the Ech cycle , the halint(property) relationships are analyzed for the homo-halogen bonds of the [(A)nY\u2013X\u00b7\u00b7\u00b7X\u2013Z(B)m] type formed upon the interaction of two neutral fragments (A)nY\u2013X and X\u2013Z(B)m . For the first time, the Eint(property) relationships were deduced for the Cl\u00b7\u00b7\u00b7Cl, Br\u00b7\u00b7\u00b7Br, and I\u00b7\u00b7\u00b7I homo-XBs based on a large, statistically significant set of structures .In this work, the Eint(Vb) correlation for structures of the [(A)nZ\u2013Y\u00b7\u00b7\u00b7F]\u2212 type at the M06-2X level of theory has similar parameters as that for the MP4, CCSD, and CCSD(T) methods .int and the electron density based properties of the [(A)nZ\u2013Y\u00b7\u00b7\u00b7F]\u2212 structures. Coefficients of the Eint(Vb) correlations obtained for these two basis sets are also similar . MeanwhThe Hessian matrix was calculated for all optimized structures to prove the location of correct minima. No symmetry operations were applied during the calculations. The stability test was performed and the stable solutions were achieved for all structures using the keyword STABLE(OPT). BSSE was corrected using the counterpoise (CP) method ,100. A dnY\u2013X\u00b7\u00b7\u00b7X\u2013Z(B)m] bearing the homo-halogen bonds Cl\u00b7\u00b7\u00b7Cl, Br\u00b7\u00b7\u00b7Br, and I\u00b7\u00b7\u00b7I and formed upon interaction of two neutral fragments (A)nY\u2013X and X\u2013Z(B)m were selected as computational models . In this work, the fragment (A)nY\u2013X (at the left side of the complex formula) corresponds to the XB donor and the fragment X\u2013Z(B)m (at the right side of the complex formula) corresponds to the XB acceptor.For this study, structures [(A)nY\u2013X\u00b7\u00b7\u00b7X\u2013F], [(A)nY\u2013X\u00b7\u00b7\u00b7X\u2013H], and [F\u2013X\u00b7\u00b7\u00b7X\u2013Z(B)m]. In the first two series, the XB donor part is variable while the XB acceptor part is fixed with the Z(B)m group being either a strong electron acceptor (Z(B)m = F) or an electron donor (Z(B)m = H). Series of the structures with even stronger electron donor groups Z(B)m = CMe3, CHMe2, or CH3 were also calculated. However, in most of cases, secondary X\u00b7\u00b7\u00b7H\u2013C interactions were found and, therefore, the individual X\u00b7\u00b7\u00b7X halogen bonds cannot be isolated for the analysis.For each type of halogen bonds, three series were considered, i.e., [(A)m], the XB acceptor part is variable whereas the XB donor part is fixed ((A)nY = F). Most of the attempts to calculate similar series with an electron donor group (A)nY = H failed because the \u03c3-hole at the terminal X atom of the XB donor is not sufficiently pronounced in this case, and other interactions prevail in the resulting structures.In the third series, type with the X\u00b7\u00b7\u00b7Z contact shorter the sum of van der Waals radii but with \u03b82 \u2265 70\u00b0 and with no other weak interactions were included in the analysis since no BCPs for the X\u00b7\u00b7\u00b7Z contacts were found). Effect of the angle \u03b82 on the correlations under study is discussed in 2\u201d.Only structures which have no other contacts shorter than the sum of van der Waals radii between the (A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m], [(A)nY\u2013Br\u00b7\u00b7\u00b7Br\u2013Z(B)m], and [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013Z(B)m] are called \u201clarge\u201d, while the series [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013H], [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013F], [F\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m], [(A)nY\u2013Br\u00b7\u00b7\u00b7Br\u2013H], [(A)nY\u2013Br\u00b7\u00b7\u00b7Br\u2013F], [F\u2013Br\u00b7\u00b7\u00b7Br\u2013Z(B)m], [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013H], [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013F], and [F\u2013I\u00b7\u00b7\u00b7I\u2013Z(B)m] are called \u201csmall\u201d series.Further in this work, the series [(A)int increases along the row X = Cl < Br < I . The Cl\u00b7\u00b7\u00b7Cl interaction is slightly weaker than the corresponding Br\u00b7\u00b7\u00b7Br and I\u00b7\u00b7\u00b7I bonds for the [(A)nY\u2013X\u00b7\u00b7\u00b7X\u2013H] and [F\u2013X\u00b7\u00b7\u00b7X\u2013Z(B)m] series (m] are the strongest ones while the XBs in the series [(A)nY\u2013X\u00b7\u00b7\u00b7X\u2013F] are typically the weakest bonds.The calculated X\u00b7\u00b7\u00b7X interaction energy with the BSSE correction in the whole set of the structures varies from 0.34 to \u22129.24 kcal/mol. All three types of halogen bonds, Cl\u00b7\u00b7\u00b7Cl, Br\u00b7\u00b7\u00b7Br, and I\u00b7\u00b7\u00b7I, have comparable strengths. The dispersion of E] series . The halint(property) relationships are considered for each estimator within the whole set and various series of the structures.In the next sections, the Eb).Potential energy density at BCP correlations reported in literature obey a linear law. The R2 values (0.80) indicate that the correlations are of rather poor quality. Meanwhile, the mean absolute deviation (MAD) for these fittings is ~0.60 kcal/mol. This value is well within the typical accuracy of the DFT methods and this is almost three times less than the average interaction energy for this structural set .ructures A. Very cnY\u2013X\u00b7\u00b7\u00b7X\u2013Z(B)m] with the homo-halogen bonds formed upon interaction of the neutral fragment, Eint may be roughly estimated from Vb using a single formula for all X = Cl, Br, and I. This situation is very different from that found recently for the set of anionic structures [(A)nZ\u2013Y\u00b7\u00b7\u00b7X]\u2212 which could describe the whole set of structures. At least two various dependencies are clearly visible on each Eint(property) plot (ty) plot D\u2013G.Potential energy density at BCP. The Eint(Vb) correlations for each specific type of the halogen bond are of significantly better quality than those for the whole set in terms of both R2 (0.92\u20130.96) and MAD (nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m] and [(A)nY\u2013Br\u00b7\u00b7\u00b7Br\u2013Z(B)m] may be well approximated by a single quadratic or exponential function, the former performing slightly better with R2 = 0.92 and MAD = 0.32 kcal/mol. The [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013Z(B)m] series lies below the series with the Cl\u00b7\u00b7\u00b7Cl and Br\u00b7\u00b7\u00b7Br bonds on the \u2212Eint(\u2013Vb) plot.cal/mol) A. The ClKinetic energy density at BCP. The kinetic energy density behaves similarly to the potential energy density for the estimate of Eint within these series with the MAD values being only slightly worse relationships for all series [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m], [(A)nY\u2013Br\u00b7\u00b7\u00b7Br\u2013Z(B)m] and [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013Z(B)m] have similar parameters despite the application of different basis sets . Meanwhile, the Eint(Vb) relationship for the iodine halogen bonds is quite different from those for the Cl\u00b7\u00b7\u00b7Cl and Br\u00b7\u00b7\u00b7Br structures. Thus, the application of Gb as an estimator of Eint is preferable over Vb also from this point of view.cal/mol) B. HoweveElectron density at BCP. The Eint(\u03c1b) relationships for these series are similar to Eint(Vb) and Eint(Gb) nY\u2013Br\u00b7\u00b7\u00b7Br\u2013Z(B)m] and [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013Z(B)m] may be approximated by a single function , while the fitting for the [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m] series has quite different parameters.Eint(Gb) C. In conLaplacian and curvature of electron density distribution at BCP. Both Eint(\u22072\u03c1b) and Eint relationships for these series have similar features and are described by either quadratic or exponential function relationships demonstrate different behavior for the Cl\u00b7\u00b7\u00b7Cl and Br\u00b7\u00b7\u00b7Br series, on one side, and for the I\u00b7\u00b7\u00b7I series, on the other side . For the Br\u00b7\u00b7\u00b7Br series, the Eint(Hb) function is not well-defined. Finally, for the I\u00b7\u00b7\u00b7I structures, \u2212Eint increases with the decrease of Hb. The \u2212Eint(Hb) relationship is nearly linear with R2 = 0.93 and MAD = 0.42 kcal/mol and may be used for the estimate of Eint for this type of halogen bonds.her side F. For thInternuclear distance d(X\u00b7\u00b7\u00b7X). The \u2212Eint(d(X\u00b7\u00b7\u00b7X)) dependencies are exponential with rather different parameters for the Cl\u00b7\u00b7\u00b7Cl, Br\u00b7\u00b7\u00b7Br, and, in particular, I\u00b7\u00b7\u00b7I structures plots. The best Eint(Gb) approximation was found for the [F\u2013X\u00b7\u00b7\u00b7X\u2013Z(B)m] series with R2 = 0.86 and MAD = 0.44 kcal/mol (int(\u03c1b) function also reasonably describes each of the pairs \u201c[(A)nY\u2013Br\u00b7\u00b7\u00b7Br\u2013H] + [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013H] and \u201c[(A)nY\u2013Br\u00b7\u00b7\u00b7Br\u2013F] + [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013F] (int(property) relationships significantly depend on the nature of the interacting atoms X.Typically, there are no statistically meaningful trends describing these series . Severalkcal/mol A. The EiI\u00b7\u00b7\u00b7I\u2013F] B but thePotential energy density at BCP. The Eint(Vb) relationship for each of nine \u201csmall\u201d series may be reasonably approximated by a linear function. The quadratic fitting is noticeably better than the linear one only for the [F\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m], [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013H], and [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013F] series (] series A\u2013C. The int(\u2212Vb) relationships for the [F\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m], [F\u2013Br\u00b7\u00b7\u00b7Br\u2013Z(B)m], and [F\u2013I\u00b7\u00b7\u00b7I\u2013Z(B)m] series are significantly higher than for the other series, whereas those for the [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013F], [(A)nY\u2013Br\u00b7\u00b7\u00b7Br\u2013F], and [(A)nY\u2013I\u00b7\u00b7\u00b7I\u2013F] structures are the lowest.First, the slope and negative interception of the \u2212E2. However, the MAD values for the \u201csmall\u201d series are significantly lower than for the corresponding \u201clarge\u201d series . The estimate of Eint for the structures [F\u2013Br\u00b7\u00b7\u00b7Br\u2013Z(B)m] and [F\u2013I\u00b7\u00b7\u00b7I\u2013Z(B)m] is preferable using equations for the \u201clarge\u201d series because both MAD and R2 parameters are worse for these \u201csmall\u201d series than for the corresponding \u201clarge\u201d ones.Second, quality of the dependencies for the \u201csmall\u201d series is usually worse than for the \u201clarge\u201d ones in terms of RnY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013H], is described by an equation similar to the EML formula (Eint = 0.47Vb + 0.27). Equations (3)\u2013(5) obtained for the halogen bonds of the X\u00b7\u00b7\u00b7D type [nY\u2013X\u00b7\u00b7\u00b7X\u2013Z(B)m] type discussed in this work (except the same [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013H] series).Third, among all \u201csmall\u201d series, only one, , [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013F] and [F\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m] relationship is of the same type for [(A)nY\u2013Br\u00b7\u00b7\u00b7Br\u2013H] and [(A)nY\u2013Br\u00b7\u00b7\u00b7Br\u2013F] m] and as a linear function for the I...I structures (int(Hb) dependencies for the \u201csmall\u201d series is worse than for the other estimators.l\u2013Z(B)m] D althoug\u00b7\u00b7\u00b7Br\u2013F] E. Howeveructures F. The quOther estimators. The main features of the Eint(Gb), Eint(\u03c1b), Eint(\u22072\u03c1b), Eint, and Eint(d(X\u00b7\u00b7\u00b7X)) relationships for the \u201csmall\u201d series are similar to the corresponding Eint(Vb) dependencies (m] particularly poor.ndencies . Laplacir) over IAS (r \u2208 IAS) may be a better estimator of Eint than the potential energy density at BCP (Vb). This result was obtained for a set of 50 structures with very different types of non-covalent interactions. Here, the quality of this estimator was verified for structures of the [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m] type. The calculations indicated that the IAS integral of electronic virial field behaves similarly as Vb for the [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013H] and [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013F] series and relatively high MAD . Correspondingly, the Eint(nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m] is also significantly worse than the Eint(Vb) relationship with R2 = 0.89 vs. 0.96 and MAD = 0.28 vs. 0.18 kcal/mol.Recently, Romanova, Lyssenko, and Ananyev reported compare A. Howeveb against nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013H] (R2 = 0.99), a quite good relationship for [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013F] (R2 = 0.96) but a poor dependence for [F\u2013Cl\u00b7\u00b7\u00b7Cl\u2013Z(B)m] (R2 = 0.75) B. Thus, 1\u00b7\u00b7\u00b7Z contact in structures [(A)nY\u2013X1\u00b7\u00b7\u00b7X2\u2013Z(B)m] included in the analysis, some of them have the angleX1X2Z (\u03b82) lower than 90\u00b0. This may point out some interaction between the atoms X1 and Z. To verify if this possible interaction affects the Eint(property) dependencies, all structures were divided into three groups, i.e., those with \u03b82 \u2265 90\u00b0, 80\u00b0 \u2264 \u03b82 < 90\u00b0, and 70\u00b0 \u2264 \u03b82 < 80\u00b0. In the second and third groups, structures with the X1\u00b7\u00b7\u00b7Z distance shorter than the sum of van der Walls radii were also separated from those with the X1\u00b7\u00b7\u00b7Z distance longer than this sum.Although no BCP was found for the Xint(Vb) dependencies for all these groups of structures are shown in 2 and the X1\u00b7\u00b7\u00b7Z distance. If considering only structures with \u03b82 \u2265 90\u00b0, R2 and MAD values are similar to those for the complete structural series except the Br\u00b7\u00b7\u00b7Br structures on the Eint(Vb) dependencies. Similar results were obtained for all other estimators.The E compare D. In theint(property) correlations were established and analyzed for the first time for the large statistically significant sets of the homo-halogen bonds Cl\u00b7\u00b7\u00b7Cl, Br\u00b7\u00b7\u00b7Br, and I\u00b7\u00b7\u00b7I formed upon interaction of two neutral fragments at the M06-2X/6-31+G* (the Cl\u00b7\u00b7\u00b7Cl and Br\u00b7\u00b7\u00b7Br bonds) and M06-2X/DZP (the I\u00b7\u00b7\u00b7I bond) levels of theory. Electron density, its Laplacian, curvature of the electron density distribution, potential, kinetic, and total energy densities at BCP, integral of electronic virial over IAS and the d(X\u00b7\u00b7\u00b7X) internuclear distance were examined as estimators of the interaction energy. The correlations obtained in this work have a significant practical potential since they lead to interaction energies of the entire classes of the halogen bonds with the electron density distribution or even only the d(X\u00b7\u00b7\u00b7X) internuclear distance being in the hands. This is particularly important for the systems with multiple intermolecular non-covalent interactions or those bearing intramolecular XBs because a direct determination or calculation of Eint for such systems is an extremely difficult or often even impossible task. The following conclusions can be made.In this work, the Eint = 0.128Gb2 \u2212 0.82Gb + 1.66 with R2 = 0.91 and MAD = 0.39 kcal/mol. For other estimators, no reasonable single correlation was found for the whole structural set.First, the whole set of structures can be reasonably approximated by a single quadratic function \u2212Eint(Vb), Eint(Gb), Eint(\u03c1b), Eint, and Eint(d(X\u00b7\u00b7\u00b7X)) relationships, all of them being nonlinear . Quality of these relationships is better for the Cl\u00b7\u00b7\u00b7Cl structures compared to the Br\u00b7\u00b7\u00b7Br and I\u00b7\u00b7\u00b7I ones. Total energy density behaves well only for the I\u00b7\u00b7\u00b7I structures while the \u2212Eint(Hb) function is not well-defined for the Br\u00b7\u00b7\u00b7Br structures. Laplacian \u22072\u03c1b works well for the Cl\u00b7\u00b7\u00b7Cl and Br\u00b7\u00b7\u00b7Br series but not for the I\u00b7\u00b7\u00b7I one.Second, each of the series Cl\u00b7\u00b7\u00b7Cl, Br\u00b7\u00b7\u00b7Br, and I\u00b7\u00b7\u00b7I can be well described individually by the Eint due to lower dispersion of the Eint(Gb) functions.Third, the kinetic energy density at BCP can be recommended as the best estimator of EnY\u2013X\u00b7\u00b7\u00b7X\u2013H], [(A)nY\u2013X\u00b7\u00b7\u00b7X\u2013F], and [F\u2013X\u00b7\u00b7\u00b7X\u2013Z(B)m].Fourth, typically, there are no statistically meaningful trends describing the series [(A)2 but it is better in terms of MAD (with exception of the [F\u2013Br\u00b7\u00b7\u00b7Br\u2013Z(B)m] and [F\u2013I\u00b7\u00b7\u00b7I\u2013Z(B)m] structures).Fifth, quality of correlations for the \u201csmall\u201d series is usually worse than that for the \u201clarge\u201d series Cl\u00b7\u00b7\u00b7Cl, Br\u00b7\u00b7\u00b7Br, and I\u00b7\u00b7\u00b7I in terms of Rint(Vb) and Eint(Gb) relationships are close to Equations (1)\u2013(5) only for the [(A)nY\u2013Cl\u00b7\u00b7\u00b7Cl\u2013H] structures. Additionally, the Eint(property) correlations obtained here for the Cl\u00b7\u00b7\u00b7Cl and Br\u00b7\u00b7\u00b7Br bonds are significantly different from those found previously for the Cl\u00b7\u00b7\u00b7Cl\u2212 and Br\u00b7\u00b7\u00b7Br\u2212 interactions between the neutral fragments (A)nY\u2013Hal and the halide anion Hal\u2212 [Sixth, the obtained here Eion Hal\u2212 . This onint estimator at least for the Cl\u00b7\u00b7\u00b7Cl bond.Seventh, the IAS integral of electronic virial Eighth, the BSSE effect is insignificant for the Cl\u00b7\u00b7\u00b7Cl structures but becomes important for the I\u00b7\u00b7\u00b7I and, in particular, Br\u00b7\u00b7\u00b7Br structures.int(property) relationships recommended for the Cl\u00b7\u00b7\u00b7Cl, Br\u00b7\u00b7\u00b7Br, and I\u00b7\u00b7\u00b7I interactions are given in i) establishment of the Eint(property) relationships for the hetero-halogen bonds Hal1\u00b7\u00b7\u00b7Hal2 and other types of the non-covalent interactions and (ii) analysis of the effect of computational method, basis set and effective core pseudopotentials on these correlations. The corresponding studies are currently underway.The E"} +{"text": "The title compound, the potassium salt of a benzo\u00adthia\u00adzol(methyl\u00adsulfan\u00adyl)pyriminidine, was obtained in a reaction designed to deliver a neutral 2-pyrimidylbenzo\u00adthia\u00adzole. It crystallized with two independent mol\u00adecular units in the asymmetric unit. +\u00b7C18H14N5O2S3\u2212\u00b7C3H7NO\u00b70.5H2O, was obtained in a reaction designed to deliver a neutral 2-pyrimidylbenzo\u00adthia\u00adzole. The anion is deprotonated at the sulfonamide nitro\u00adgen. The asymmetric unit of the title compound contains two potassium cations, two anions, two mol\u00adecules of DMF and one of water. The anions display some conformational differences but each contains an intra\u00admolecular N\u2014H\u22efNbenzo\u00adthia\u00adzole hydrogen bond. The potassium ions both display a highly irregular six-coordination, different for each potassium ion. The anions, together with the DMF and water mol\u00adecules, are linked by four classical hydrogen bonds to form chains parallel to the b-axis direction.The title compound, K The atom K2 is coordinated by N2\u2032, N5\u2032 and both DMF oxygen atoms within the asymmetric unit, plus O2 at and O1 at . The angles subtended by the chelating anions via N2/N5 are particularly narrow. The bridging nature of O92 is shown in Fig.\u00a03The potassium ions both display a highly irregular six-coordination; all K\u2014N and K\u2014O contacts Table\u00a01 are < 2.S(6) ring motif. The anions display some differences in conformation; the angle between the benzo\u00adthia\u00adzole ring (seven atoms) and the pyrimidine ring plus immediate substituents (ten atoms) is 20.56\u2005(5)\u00b0 for anion 1 (unprimed atoms) but 42.20\u2005(2)\u00b0 for anion 2 (primed atoms). Comparing the torsion angles in Table\u00a01cf. torsion angle C10\u2014N5\u2014S3\u2014C13 is 62.49\u2005(11)\u00b0, and 65.28\u2005(11)\u00b0 in the non-inverted system.Each anion displays an intra\u00admolecular hydrogen bond -6-(methyl\u00adsulfan\u00adyl)pyrimidin-2-yl](phenyl\u00adsulfon\u00adyl)aza\u00adnide di\u00admeth\u00adyl\u00adformamide monosolvate hemihydrate (5):d]thia\u00adzole-2-yl)-3,3-bis\u00ad(methyl\u00adthio)\u00adacrylo\u00adnitrile (2) (0.01\u2005mol) was added to a stirred solution of the N-(di\u00adamino\u00admethyl\u00adene)benzene\u00adsulfonamide (3) (0.01\u2005mol) in dry dioxane (20\u2005ml) containing potassium hydroxide (0.01\u2005mol); the reaction mixture was refluxed for 2\u2005h. After completion of the reaction (TLC), the solid precipitate was filtered off, and then recrystallized from DMF/H2O to give colourless block-like crystals of compound 5, the potassium salt of compound 4, in 75% yield (m.p. = 517\u2005K). IR : \u03bd 3431 and 3874 . 1H NMR : \u03b4 2.19 , 7.32\u20137.39 , 7.46 , 7.83\u20137.85 , 7.92 , 8.01 , 8.49 , 11.50 .The reaction pathway is illustrated in Fig.\u00a01Uiso(H) = 1.5Ueq(C-meth\u00adyl)], and allowed to rotate but not to tip (AFIX 137). Other hydrogen atoms were included using a riding model starting from calculated positions: C\u2014Haromatic = 0.95\u2005\u00c5 with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019002275/su5477sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019002275/su5477Isup2.hklStructure factors: contains datablock(s) I. DOI: 1896740CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dihedral angles between the aromatic rings are 31.99\u2005(3) and 9.17\u2005(5)\u00b0 for 1 and 2, respectively. Compound 1 features an intra\u00admolecular bifurcated N\u2014H\u22ef link due to the presence of the ortho-Cl atom on the benzene ring, whereas 2 features an intra\u00admolecular N\u2014H\u22efO hydrogen bond. In the crystal of 1, inversion dimers linked by pairs of N\u2014H\u22efS hydrogen bonds generate R22(8) loops. The extended structure of 2 features the same motif but an additional weak C\u2014H\u22efS inter\u00adaction links the inversion dimers into [100] double columns. Hirshfeld surface analyses indicate that the most important contributors towards the crystal packing are H\u22efH (26.6%), S\u22efH/H.\u00b7S (13.8%) and Cl\u22efH/H\u22efCl (9.5%) contacts for 1 and H\u22efH (19.7%), C\u22efH/H\u22efC (14.8%) and Br\u22efH/H\u22efBr (12.4%) contacts for 2.The title compounds, C The biological activities of these compounds were previously reported by Khan et al. \u2005\u00c5] benzoyl ring linked to a ortho-chloro-substituted phenyl ring [C\u2014Cl = 1.7387\u2005(14)\u2005\u00c5] in while in 2 \u2005\u00c5] benzoyl ring is linked to a para-bromo-substituted phenyl ring [C\u2014Br = 1.8991\u2005(17)\u2005\u00c5] via a thio\u00adurea (S1/N1/N2/C8) linkage. The benzoyl (O1/C1\u2013C7) and phenyl rings (C9\u2013C14) are arranged about the thio\u00adurea moiety in an anti fashion having torsion angles C8\u2014N1\u2014C7\u2014C6 = \u2212170.22\u2005(13) and C9\u2014N2\u2014C8\u2014S1 = 4.5\u2005(2)\u00b0 in compound 1, with corresponding values of \u2212176.01\u2005(16) and 3.8\u2005(3)\u00b0, respectively, in compound 2. The dihedral angles between the phenyl rings are 31.99\u2005(3) and 9.17\u2005(5)\u00b0 in 1 and 2, respectively. Compound 1 features an intra\u00admoleclar bifurcated N\u2014H\u22ef hydrogen bond (Table\u00a01ortho-Cl atom whereas 2 has an intra\u00admolecular N\u2014H\u22efO link (Table\u00a02S(6) ring. These intra\u00admolecular hydrogen bonds may be responsible for the anti arrangement of the aromatic rings about the thio\u00adurea linker.Compound 1 Fig.\u00a01 is compo 2 Fig.\u00a02, a para-d Table\u00a01 due to tk Table\u00a02. Both st1, inversion dimers linked by pairwise N1\u2014H1A\u22efS1 hydrogen bonds phen\u00adyl]carbamo\u00adthio\u00adyl}-3-methyl\u00adbenzamide (CCDC deposition No. 1840069) and 4-chloro-N-{[4-chloro-3-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl]carbamo\u00adthio\u00adyl}benzamide (CCDC 1587395) for compounds related to 1 and 2, Hirshfeld surfaces (26.6%) contacts, followed by S\u22efH/H\u22efS (13.8%), Cl\u22efH/H\u22efCl (9.5%) O\u22efH/H\u22efO (6.7%), F\u22efH/H\u22efF (6.6%), Cl\u22efF/F\u22efCl (3.7%) and F\u22efC/C\u22efF (3.1%) inter\u00adactions. In compound 2, H\u22efH (19.7%) are the most significant, followed by C\u22efH/H\u22efC (14.8%), S\u22efH/H\u22efS (12.6%), Br\u22efH/H\u22efBr (12.4%), C\u22efC (9.9%) and O\u22efN/N\u22efO (7.9%) inter\u00adactions.In order to further analyse the close contacts and inter\u00admolecular inter\u00adactions in the crystals of x) Fig.\u00a05 and two-x) Fig.\u00a05 and 7 \u25b8 1 and 2 were synthesized by adopting a literature procedure or 4-bromo\u00adphenyl aniline (for 2) were added to the mixture and refluxed at 343\u2005K. Hydro\u00adchloric acid was added and the solution was filtered to obtain the desired products: 1 in 69% yield and 2 in 80% yield. For recrystallization, compound 1 was dissolved in a mixture of di\u00adchloro\u00admethane and methanol (1:1) while compound 2 was dissolved in di\u00adchloro\u00admethane and left for slow evaporation at room temperature to obtain colourless prisms of 1 and colourless plates of 2Compounds 1, the N-bound H atoms were located in difference-Fourier maps and their positions were freely refined; in 2, the N-bound H atoms were located in difference-Fourier maps and refined as riding atoms in their as-found relative positions. The constraint Uiso(H) = 1.2Ueq(carrier) was applied in all cases.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019008569/hb7804sup1.cifCrystal structure: contains datablock(s) global, 1, 2. DOI: Click here for additional data file.10.1107/S2056989019008569/hb78041sup4.cmlSupporting information file. DOI: 10.1107/S2056989019008569/hb78041sup4.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989019008569/hb78042sup3.hklStructure factors: contains datablock(s) 2. DOI: Click here for additional data file.10.1107/S2056989019008569/hb78042sup5.cmlSupporting information file. DOI: 1923234, 1923233CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, extensive hydrogen bonding, with the di\u00admethyl\u00adammonium cation and the water mol\u00adecule linking the complex anions, results in the formation of a supra\u00admolecular framework.The title complex was synthesized sonochemically. The ligand was formed 3)2NH2][Cr(C6H10NO4)2Cl]\u00b7H2O, was synthesized sonochemically. The complex anion consists of a chromium(II) ion ligated by two 3-carb\u00adoxy-2-(di\u00admethyl\u00adamino)\u00adpropano\u00adate anions. They coordinate in a bidentate manner, with a carboxyl\u00adate oxygen atom and the nitro\u00adgen atom cis to each other in the equatorial plane, while the apical position is occupied by a Cl\u2212 ion. Hence, the chromium(II) ion is five-coordinate with a quasi-ideal square-pyramidal geometry; \u03c45 parameter = 0.01. The complex crystallizes as a monohydrate and in the crystal, the water mol\u00adecule and the di\u00admethyl\u00adammonium counter-ion link the complex cations via N\u2014H\u22efO, N\u2014H\u22efCl, Owater\u2014H\u22efO, O\u2014H\u22efOwater and O\u2014H\u22efO hydrogen bonds, forming a supra\u00admolecular framework. There are also a number of C\u2014H\u22efO hydrogen bonds present that reinforce the framework structure. The crystal studied was refined as a racemic twin.The title complex, [(CH The chloride anion, Cl1, occupies the apical position. The five-coordinate chromium ion is displaced by 0.3469\u2005(7)\u2005\u00c5 from the mean plane through atoms O1, N1, O5 and N2. The equatorial Cu\u2014O bond lengths are Cr1\u2014O1 = 1.960\u2005(5)\u2005\u00c5 and Cr1\u2014O5 = 1.954\u2005(5)\u2005\u00c5, while the equatorial Cu\u2014N bond lengths are slightly longer viz. Cr1\u2014N1 = 2.025\u2005(5)\u2005\u00c5 and Cr1\u2014N2 = 2.030\u2005(5)\u2005\u00c5. The axial Cr1\u2014Cl1 bond length is 2.5301\u2005(16)\u2005\u00c5. The C\u2014C, C\u2014O, and C\u2014N bond lengths of the ligands are close to those reported for similar compounds to 100.88\u2005(16)\u00b0 and from 159.6\u2005(2) to 160.3\u2005(2)\u00b0, respectively. This leads to a quasi-ideal square-pyramidal geometry for atom Cr1 with a \u03c45 parameter of 0.01 -dimethyl 3-(di\u00adphenyl\u00adphosphino)-N,N-di\u00admethyl\u00adaspartate]di\u00adchloro\u00adpalla\u00addium(II) in 10\u2005ml of water was added with magnetic stirring for a further 30\u2005min. The mixture was then put in an ultrasonic bath for 2h. The solution was then left to evaporate slowly and blue prismatic crystals were collected after two months.A mixture of fumaric acid and di\u00admethyl\u00adamine hydro\u00adchloride (0.09\u2005ml) dissolved in 20\u2005ml methanol was stirred for 1\u2005h. Chromium(II) acetate dihydrate [CrUiso(H) = 1.5Ueq(O). All other H atoms were placed in geometrically idealized positions and constrained to ride on their parent atoms: O\u2014H = 0.82\u2005\u00c5, N\u2014H = 0.89\u2005\u00c5, C\u2014H = 0.96\u20130.99\u2005\u00c5 with Uiso(H) = 1.5Ueq and 1.2Ueq for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019004717/su5493sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989019004717/su5493Isup2.hklStructure factors: contains datablock(s) I. DOI: 1892216CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound is a host\u2013guest complex of the tris-urea receptor, 3-(4-nitro\u00adphen\u00adyl)-1,1-bis\u00ad{2-[3-(4-nitro\u00adphen\u00adyl)ureido]eth\u00adyl}urea, encapsulating a hydrogen-bonded chain of di\u00adhydrogen phosphate anions. 25H25N9O9\u00b7C16H36N+\u00b7H2PO4\u2212 (I) or (C25H25N9O9)\u00b7(n-Bu4N+)\u00b7(H2PO4\u2212) (systematic name: 3-(4-nitro\u00adphen\u00adyl)-1,1-bis\u00ad{2-[3-(4-nitro\u00adphen\u00adyl)ureido]eth\u00adyl}urea tetra\u00adbutyl\u00adammonium di\u00adhydrogen phosphate), comprises a tris-urea receptor (R), a di\u00adhydrogen phosphate anion and a tetra-n-butyl\u00adammonium cation. It crystallizes with two independent formula units in the asymmetric unit. The conformations of the two tris-urea receptors are stabilized by N\u2014H\u22efO and C\u2014H\u22efO intra\u00admolecular hydrogen bonds. Each di\u00adhydrogen phosphate anion has two O\u2014H\u22efO inter\u00admolecular hydrogen-bonding inter\u00adactions with the other di\u00adhydrogen phosphate anion. Inversion-related di-anion units are linked by further O\u2014H\u22efO hydrogen bonds, forming a chain propagating along the a-axis direction. Each di\u00adhydrogen phosphate anion makes a total of four N\u2014H\u22efO(H2PO4\u2212) hydrogen bonds with two ureido subunits from two different tris-urea receptors, hence each tris-urea receptor provides the two ureido subunits for the encapsulation of the H2PO4\u2212 hydrogen-bonded chain. There are numerous inter\u00admolecular C\u2014H\u22efO hydrogen bonds present involving both receptor mol\u00adecules and the tetra-n-butyl\u00adammonium cations, so forming a supra\u00admolecular three-dimensional structure. One of the butyl groups and one of the nitro groups are disordered over two positions of equal occupancy.The title compound, C Inter\u00adestingly, a one-dimensional hydrogen-bonded polymeric structure is formed via hydrogen bonds between di\u00adhydrogen phosphate anions, and this anionic polymer is surrounded by and linked to the tris-urea receptors through ureido N\u2014H\u22efO hydrogen bonds.Anions play an important role in many chemical, catalysis, environmental and biological systems (Sessler R1 and R2) and the di\u00adhydrogen phosphate anions (P1 and P2) are illustrated in Fig.\u00a01N\u22efO7 in R1 and N17\u2014H17N\u22efO16 in R2), each forming an S(9) ring motif,. Four intra\u00admolecular C\u2014H\u22efO hydrogen bonds are also present in receptor R1 and three in R2 . The urea subunits N5/N6 in R1 and N11/N12 in R2 are orientated towards the di\u00adhydrogen phosphate ions (P1 and P2) forming a 2:2 adduct via N\u2014H\u22efO hydrogen bonds with anion P1 and enclosing trans to the C=O group across the respective C\u2014N bond. Anions P1 and P2 inter\u00adact with each other via two O\u2014H\u22efO hydrogen bonds (O20\u2014H20O\u22efO24 and O23\u2014H23O\u22efO22), enclosing an R1 and from 6.87\u2005(14) to 13.82\u2005(14)\u00b0 in R2. The dihedral angle between the nitro group and the benzene ring to which it is attached also vary, from 7.1\u2005(3) to 13.4\u2005(4)\u00b0 in R1 and from 8.3\u2005(4) to 16.7\u2005(7)\u00b0 in R2.The title compound crystallizes with two independent formula units in the asymmetric unit. The mol\u00adecular structure of the two tris-urea receptors , that are linked by O\u2014H\u22efO hydrogen bonds O20\u2014H20O\u22efO24 and O23\u2014H23O\u22efO22, are further linked to inversion-related anions via hydrogen bonds O21\u2014H21O\u22efO19i and O25\u2014H25O\u22efO26ii amine (tren). One of these compounds, tetra-n-butyl\u00adammonium tris\u00ad(2-(N-perfluoro\u00adphenyl\u00adureaylato)eth\u00adyl)amine di\u00adhydrogen phosphate di\u00admethyl\u00adformamide monosolvate, encapsulates a dimer of H2PO4\u2212 anions forming a pseudo-dimeric cage via sixteen hydrogen bonds and two weak anion\u22ef\u03c0 inter\u00adactions ]tris\u00ad(N\u2032-phenyl\u00adurea) tetra-n-butyl\u00adammonium di\u00adhydrogen phosphate bis\u00ad(di\u00adhydrogen phosphate) -1-(4-nitro\u00adphen\u00adyl)-3-{2-[3-(4-nitro\u00adphen\u00adyl)ureido]cyclo\u00adhex\u00adyl}urea aceto\u00adnitrile monosolvate \u20132.21\u2005(2)\u2005\u00c5 in the title compound. The O\u2014H\u22efO hydrogen bonds involving the H2PO4\u2212 anions are also very similar: 1.62\u2005(3) and 1.63\u2005(4)\u2005\u00c5 in DASNUH, while they vary from 1.59\u2005(2) to 1.66\u2005(2)\u2005\u00c5 in the title complex.Chiral anion receptors with two enanti\u00adomeric forms Synthesis of 3-(4-nitro\u00adphen\u00adyl)-1,1-bis\u00ad{2-[3-(4-nitro\u00adphen\u00adyl)ureido]eth\u00adyl}urea (R): In a 250\u2005ml round-bottom flask, di\u00adethyl\u00adenetri\u00adamine dissolved in 100\u2005ml of dry CH2Cl2 was added dropwise under vigorous stirring to a solution of 20\u2005ml of dry CH2Cl2 containing p-nitro\u00adbenzene iso\u00adcyanate . Subsequently, the reaction mixture was allowed to reflux for 24\u2005h. A yellowish solid was collected by filtration and washed using sequentially CH2Cl2 (3 \u00d7 70\u2005ml), a solvent mixture and diethyl ether (3 \u00d7 70\u2005ml). The solid was then dried in vacuo overnight to afford the receptor R as a light-brown powder . FT\u2013IR : 3339, 1679, 1606, 1559, 1501, 1330. 1H NMR in ppm: \u03b4 = 9.42 , 9.23 , 8.10 , 7.76 , 7.59 , 6.60 , 3.51 . 13C NMR in ppm: \u03b4 = 154.99, 154.67, 147.34, 146.94, 140.72, 140.48, 125.09, 124.70, 118.13, 116.95, 46.63, 40.15\u201338.89, 38.19. HRMS (ESI+): calculated for C25H25N9O9Na [M + Na]+ 618.1673 found 618.1678.Synthesis of the title complex (I) Tetra-n-butyl\u00adammonium di\u00adhydrogen phosphate (1.68\u2005mmol) was added to 5\u2005ml of a DMF solution of R (0.168\u2005mmol) and the mixture was stirred for 2\u2005h. After filtration the solution was left to evaporate slowly and yielded colourless prismatic crystals of the title complex within three weeks.Uiso(H) = 1.5Ueq(O) and 1.2Ueq(N). The C-bound H atoms were positioned geometrically and refined using a riding model: C\u2014H = 0.95\u20130.99\u2005\u00c5, with Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a02A/C79B, C80A/C80B, C81A/C81B, and C82A/C82B atoms of the disordered fragment were restrained to be similar is disordered over two positions with equal occupancies. The C\u2014C distances were refined with the restraint of 1.515\u2005(4)\u2005\u00c5. The displacement parameters of the C7910.1107/S2056989019001336/kq2020sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989019001336/kq2020Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019001336/kq2020Isup3.cmlSupporting information file. DOI: 1893140CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked 15H12F3NO3, crystallizes with one mol\u00adecule in the asymmetric unit. The mean planes of the two phenyl rings of the Schiff base moiety, bearing the OH groups and the imine group, respectively, are inclined to each other by 4.91\u2005(1)\u00b0. In the crystal, mol\u00adecules are linked via pairs of bifurcated O\u2014H\u22efO hydrogen bonds between the phenol OH groups, forming inversion dimers with an R12(5) ring motif. The structure exhibits also intra\u00admolecular O\u2014H\u22efN and C\u2014H\u22efF hydrogen-bonding inter\u00adactions. Hirshfeld surfaces analysis and two-dimensional fingerprint plots were applied to qu\u00adantify the inter\u00admolecular inter\u00adactions. The three F atoms of the tri\u00adfluoro\u00admethyl group are disordered over two sets of sites, with occupancy factors of 0.578\u2005(8) and 0.422\u2005(8). The crystal studied was refined as an inversion twinThe title compound, C The synthesis involves an aromatic amine and an aldehyde -3-({[3-meth\u00adoxy-5-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl]imino}\u00admeth\u00adyl)benzene-1,2-diol, was obtained in crystalline form from the reaction of 2,3-di\u00adhydroxy\u00adbenzaldehyde with 3-meth\u00adoxy-5-(tri\u00adfluoro\u00admeth\u00adyl)aniline.In general, Schiff bases display two possible tautomeric forms, T1(C1\u2014C6\u2014C7\u2014N1) and T2(C7\u2014N1\u2014C8\u2014C9). The respective values of = 2.0\u2005(10) and \u22125.5\u2005(11)\u00b0 indicate that the mol\u00adecule is not planar ring motif, characteristic of o-hy\u00addroxy\u00adsalicyl\u00adidene systems. Other intra\u00admolecular hydrogen bonding inter\u00adactions involve the disordered \u2013CF3 group and adjacent aromatic H atoms bonded to C9 and C11 \u2005\u00c5] and the single bond character of O1\u2014C2 [1.368\u2005(6)\u2005\u00c5] in the phenol\u2013imine tautomer. These values and other bond lengths and angles Table\u00a01 are in g1 Table\u00a02. As a reBetween adjacent mol\u00adecules there are bifurcated inter\u00admolecular O1\u2014H\u22ef hydrogen bonds with an if Fig.\u00a02, leadinget al., 2016Z)-1-phenyl-N-[3-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl]methanimine skeleton yielded eight matches. Distinctive bond lengths in the Schiff base structure are the same within standard uncertainties as those of the corresponding bond lengths in the structures of 4N--3-meth\u00adoxy\u00adsalicylaldimine -3-{[3-(tri\u00adfluoro\u00admeth\u00adyl)phenyl\u00adimino]\u00admeth\u00adyl}benzene-1,2-diol -5-(tri\u00adfluoro\u00admeth\u00adyl)aniline -2-methyl-6-[3-(tri\u00adfluoro\u00admeth\u00adyl)-phen\u00adyl\u00adimino\u00admeth\u00adyl]phenol -2-[(4-chloro\u00adphen\u00adyl)imino\u00admeth\u00adyl]-4-(tri\u00adfluoro\u00admeth\u00adoxy)phenol -4-methyl-2-[3-(tri\u00adfluoro\u00admeth\u00adyl)phen\u00adyl\u00adimino\u00admeth\u00adyl]phenol plot with de and di for the title compound is shown in Fig.\u00a03de = di = 1.5\u2005\u00c5; Fig.\u00a04Hirshfeld surface analysis of the title compound was performed utilizing the The title compound was prepared by refluxing mixed solutions of 2,3-di\u00adhydroxy\u00adbenzaldehyde in ethanol (15\u2005ml) and 3-meth\u00adoxy-5-(tri\u00adfluoro\u00admeth\u00adyl)aniline in ethanol (15\u2005ml). The reaction mixture was stirred for 5\u2005h under reflux. Single crystals of the title compound for X-ray analysis were obtained by slow evaporation of an ethanol solution .Uiso(H) set to 1.2\u20131.5Ueq(C), and with O\u2014H = 0.82\u2005\u00c5 and Uiso(H) = 1.5Ueq(O). The three F atoms of the tri\u00adfluoro\u00admethyl group are disordered over two sets of sites, with occupancy factors of 0.578\u2005(8) for F atoms with suffix A and 0.422\u2005(8) for those with suffix B I, global. DOI: 10.1107/S2056989019003220/wm5488Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019003220/wm5488Isup3.cmlSupporting information file. DOI: 1892713CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "L as the dihedral angle between the pyridyl rings is 78.37\u2005(5)\u00b0. Linear supra\u00admolecular chains are found in the crystal mediated by weak carbonyl-C=O\u22ef\u03c0(triazol\u00adyl) inter\u00adactions.The title mol\u00adecule adopts the shape of the letter 16H15N5O2, adopts the shape of the letter L with the dihedral angle between the outer pyridyl rings being 78.37\u2005(5)\u00b0; the dihedral angles between the central pyrazolyl ring (r.m.s. deviation = 0.0023\u2005\u00c5) and the methyl\u00adene-bound pyridyl and methyoxypyridyl rings are 77.68\u2005(5) and 7.84\u2005(10)\u00b0, respectively. Intra\u00admolecular amide-N\u2014H\u22efN(pyrazol\u00adyl) and pyridyl-C\u2014H\u22efO(amide) inter\u00adactions are evident and these preclude the participation of the amide-N\u2014H and O atoms in inter\u00admolecular inter\u00adactions. The most notable feature of the mol\u00adecular packing is the formation of linear supra\u00admolecular chains aligned along the b-axis direction mediated by weak carbonyl-C=O\u22ef\u03c0(triazol\u00adyl) inter\u00adactions. An analysis of the calculated Hirshfeld surfaces point to the importance of H\u22efH (46.4%), C\u22efH (22.4%), O\u22efH (11.9%) and N\u22efH (11.1%) contacts in the crystal.The title compound, C The synthesis of amide bonds utilizing T3P offers high yields, low epimerization and avoids the use of haza\u00adrdous additives such as explosive hy\u00addroxy\u00adbenzotriazole (HOBt). Further, reactions occur with high yields and lead to the easy removal of the by-products with a simple work-up, overall resulting in the formation of high-quality product. In addition, it is noted that the T3P reagent is non-toxic and non-allergenic a, these lead to linear supra\u00admolecular chains aligned along the b-axis direction. The supra\u00admolecular chains pack without specific inter\u00adactions between them, Fig.\u00a02b.The mol\u00adecular packing of (I)et al., 2016dnorm in Fig.\u00a03B, pyridyl-N3 and pyridyl-H5 atoms are indicative of short inter\u00adatomic N\u22efH/H\u22efN contacts s Table\u00a02. In addis Table\u00a02 are highs Table\u00a03 between a, and those delineated into H\u22efH, N\u22efH/H\u22efN, O\u22efH/H\u22efO, C\u22efH/H\u22efC and C\u22efC contacts de + di \u223c 2.5\u2005\u00c5 and 2.7\u2005\u00c5, respectively, in the corresponding delineated fingerprint plots Fig.\u00a05c and e. The pair of spikes with the tips at de + di \u223c 2.6\u2005\u00c5 and the regions of green points aligned in the fingerprint plot delineated into N\u22efH/H\u22efN contacts, Fig.\u00a05d, are indicative of short N\u22efH inter\u00adatomic contacts although the contribution from these contacts is relatively small. The notable percentage contributions from O\u22efN/N\u22efO and C\u22efO/O\u22efC contacts to the Hirshfeld surfaces s Table\u00a02. In the ms Fig.\u00a03b althous Table\u00a02 in the cet al., 2016The 1,3 N\u2014C and C\u2014C(=O)N(H)\u2014C substitution pattern observed in (I)H-Pyrazole-4-carb\u00adoxy\u00adlic acid (0.0446\u2005mol) was treated with diiso\u00adpropyl\u00adethyl amine (0.0669\u2005mol) and 1-propane phospho\u00adnic acid (T3P) (0.0669\u2005mol) in dimethyl formamide (10\u2005ml) at 273\u2005K for 15\u2005min. Then, 6-meth\u00adoxy\u00adpyridin-2-amine (0.0490\u2005mol) was added at 273\u2005K. The reaction mixture was heated at 353\u2005K for 3\u2005h. After completion of the reaction, the product was extracted with ethyl acetate and the excess solvent was removed under vacuum. The product was recrystallized using methanol as solvent to yield 1-(6-meth\u00adoxy\u00adpyridin-2-ylmeth\u00adyl)-1H-pyrazole-4-carb\u00adoxy\u00adlic acid. This product (0.0246\u2005mol) and 2-(chloro\u00admeth\u00adyl)pyridine (0.0295\u2005mol) were dissolved in acetone (10\u2005ml), potassium carbonate (0.0369) was added and the reaction mixture was heated at 329\u2005K for 5\u2005h. After completion of the reaction, the product was extracted with ethyl acetate twice (5\u2005ml) and the extract was concentrated under vacuum. The product was washed with diethyl ether (3\u2005ml) and recrystallized from methanol solution to obtain the title compound, (I)1Uiso(H) set to 1.2\u20131.5Ueq(C). The N-bound H atoms was refined with a distance restraint of 0.86\u00b10.01\u2005\u00c5, and with Uiso(H) = 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989018011477/hb7767sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018011477/hb7767Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018011477/hb7767Isup3.cmlSupporting information file. DOI: 1861657CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Supra\u00admolecular chains arise in the crystal as a result of O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonding.The 17H17F6N2O+\u00b7C2ClF2O3\u2212 quinolin-4-yl](hy\u00addroxy)meth\u00adyl}piperidin-1-ium chloro\u00addifluoro\u00adacetate), the cation, which is protonated at the piperidine N atom, has the shape of the letter, L, with the piperidin-1-ium group being approximately orthogonal to the quinolinyl residue [the Cq\u2014Cm\u2014Cm\u2013Na torsion angle is 177.79\u2005(18)\u00b0]. An intra\u00admolecular, charge-assisted ammonium-N\u2014H\u22efO(hydrox\u00adyl) hydrogen bond ensures the hy\u00addroxy-O and ammonium-N atoms lie to the same side of the mol\u00adecule [Oh\u2014Cm\u2014Cm\u2014Na (h = hydrox\u00adyl) = \u221259.7\u2005(2)\u00b0]. In the crystal, charge-assisted hydroxyl-O\u2014H\u22efO\u2212(carboxyl\u00adate) and ammonium-N+\u2014H\u22efO\u2212(carboxyl\u00adate) hydrogen bonds generate a supra\u00admolecular chain along [010]; the chain is consolidated by C\u2014H\u22efO inter\u00adactions. Links between chains to form supra\u00admolecular layers are of the type C\u2014Cl\u22ef\u03c0(quinolinyl-C6) and the layers thus formed stack along the a-axis direction without directional inter\u00adactions between them. The analysis of the calculated Hirshfeld surface points to the dominance of F\u22efH contacts to the surface (40.8%) with significant contributions from F\u22efF (10.5%) and C\u22efF (7.0%) contacts.In the racemic title mol\u00adecular salt, C R*,S*)-quinolin-4-yl](hy\u00addroxy\u00admeth\u00adyl)piperidin-1-ium chloride is an anti-malarial drug, being effective against the causative agent, Plasmodium falciparum . In a continuation of structural studies of Mefloquine derivatives quinolin-4-yl]-piperidin-2-yl\u00admethanol). This arises when the racemic compound is reacted with HCl: the resulting salt, [-erythro-mefloquinium] isomer. However, it should be noted that the centrosymmetric unit cell has equal numbers of the other S-,R- enanti\u00adomer, indicating that no resolution occurred during the crystallization experiment as has been observed in some of the earlier studies . The pattern of hydrogen-bonding inter\u00adactions involving the ammonium-N\u2014H H atoms (see Supra\u00admolecular features) provides confirmation of protonation at the N2 atom during crystallization and, therefore, the formation of a piperidin-1-ium cation. At the same time, delocalization of the \u03c0-electron density over the carboxyl\u00adate residue is confirmed by the equivalence of the C18\u2014O2, O3 bond lengths, i.e. 2 \u00d7 1.238\u2005(3)\u2005\u00c5.The ions comprising the asymmetric unit of (I)cf. with that of 177.79\u2005(18)\u00b0 for C3\u2014C12\u2014C13\u2014N2. The latter angle indicates the piperidin-1-ium residue is almost perpendicular to the quinolinyl residue with the methyl\u00adene-C17 group orientated towards the fused-ring system as seen in the gauche C3\u2014C12\u2014C13\u2014C17 torsion angle of \u221260.7\u2005(3)\u00b0. The observed conformation, whereby the hy\u00addroxy-O and ammonium-N atoms lie to the same side of the mol\u00adecule [the O1\u2014C12\u2014C13\u2014N2 torsion angle is \u221259.7\u2005(2)\u00b0], is stabilized by an intra\u00admolecular, charge-assisted ammonium-N2+\u2014H\u22efO1(hydrox\u00adyl) hydrogen bond, Table\u00a01L.The quinolinyl residue is not strictly planar with the r.m.s. deviation for the ten fitted non-H atoms being 0.0399\u2005\u00c5. This is also reflected in the dihedral angle formed between the and (C4\u2013C9) rings of 3.95\u2005(15)\u2005\u00c5. This aspect of the structure notwithstanding, the hydroxyl-O and ammonium-N atoms lie to opposite sides of the plane through the quinolinyl residue. This is seen in the value of the C2\u2014C3\u2014C12\u2014O1 torsion angle of \u221220.3\u2005(3)\u00b0 2C2 plane [r.m.s. deviation = 0.0089\u2005\u00c5], with the O2\u2014C18\u2014C19\u2014Cl1 torsion angle being \u221293.3\u2005(2)\u00b0, and the F7 and F8 atoms lying to the other side, the O2\u2014C18\u2014C19\u2014F7, F8 torsion angles = 28.8\u2005(3) and 146.3\u2005(2)\u00b0, respectively. The conformation of the CClF2 residue in (I)et al., 1996e.g. with mono-protonated 1,4-di\u00adaza\u00adbicyclo\u00ad[2.2.2]octane (dabco), i.e. 4-aza-1-azoniabi\u00adcyclo\u00ad[2.2.2]octane, for which three independent ion pairs comprise the asymmetric unit \u2212(carb\u00adoxyl\u00adate) and ammonium-N+\u2014H\u22efO\u2212(carboxyl\u00adate) hydrogen bonding features prominently in the mol\u00adecular packing of (I)b-axis direction, Fig.\u00a01a and Table\u00a01+\u2014H\u22efO\u2212(carboxyl\u00adate) hydrogen bonds link two cations and two anions about a centre of inversion to form eight-membered {\u22efHNH\u22efO}2 synthons, Fig.\u00a02b. These are linked into a supra\u00admolecular chain via hydroxyl-O\u2014H\u22efO\u2212(carboxyl\u00adate) hydrogen bonding, which leads to 18-membered {\u22efOCO\u22efHNC2OH}2 synthons, Fig.\u00a02b. In this scheme, the carboxyl\u00adate-O2 atom forms two hydrogen bonds. Additional stability to the supra\u00admolecular chain is afforded by quinolinyl-C\u2014H\u22efO(carboxyl\u00adate) and methyl\u00adene-C\u2014H\u22efO(hydrox\u00adyl) inter\u00adactions, Table\u00a01via C\u2014Cl\u22ef\u03c0(C4\u2013C9) inter\u00adactions, Table\u00a01a-axis direction without directional inter\u00adactions between them, Fig.\u00a02c.The presence of charge-assisted hydroxyl-O\u2014H\u22efOet al., 2016dnorm in Fig.\u00a03a and b (labelled 1\u20133), corresponding to inter\u00admolecular O\u2014H\u22efO, N\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions, Table\u00a01c and d, indicate the influence of short inter\u00adatomic H\u22efH, F\u22efH/H\u22efF and F\u22efF contacts [Table\u00a02CrystalExplorer3.1 et al., 2007i.e. 11.9%, contribution from H\u22efH contacts to the Hirshfeld surface. Conversely, the relative high number of fluorine atoms lying on the surfaces of both the cation and anion, largely participating in F\u22efH contacts, gives rise to their providing the greatest contribution, i.e. 40.8%, to the surface.The overall two-dimensional fingerprint plot and those delineated and 3.0\u2005\u00c5 (Cl\u22efH), and an arrow-shaped tip at de + di \u223c2.8\u2005\u00c5 in the fingerprint plots delineated into F\u22efH/H\u22efF, Cl\u22efH/H\u22efCl and F\u22efF contacts, respectively. The involvement of chloride and fluoride atoms in C-halogen\u22ef\u03c0 contacts, Table\u00a01In the fingerprint plot delineated into H\u22efH contacts in Fig.\u00a06et al., 2016et al., 2016et al., 2016et al., 2016i.e. (+)-PhC(CF3)(OMe)CO2\u2212 \u03b4: 1.20\u20131.35 , 1.55\u20131.75 , 3.04 , 3.53 , 5.90 , 6.94 , 8.01 , 8.13 , 8.42 , 8.72 , 9.48 ; N\u2014H H not observed. 13C NMR (DMSO-d6) \u03b4: 21.43 (2\u00d7), 21.59, 44.51, 58.90, 67.85, 1135.50. 121.17 , 121.21 , 123.64 , 126.37, 127.93 , 128.32, 128.68. 129.9 , 142.78, 146.73 , 150.97, 159.82 . 19F NMR (DMSO-d6) \u03b4: \u221258.65, \u221258.84, \u221266.68. IR (cm\u22121) 3300\u20132400 , 1662 (s).A solution of mefloquinium chloride (1\u2005mmol) and sodium di\u00adfluoro\u00adchoro\u00adacetate (1\u2005mmol) in EtOH (10ml) was refluxed for 20 mins. The reaction mixture was left at room temperature and after two days, colourless crystals of the title salt, (I)Uiso(H) set to 1.2Ueq(C). The O- and N-bound H atoms were refined with the distance restraints O\u2014H = 0.84\u00b10.01 and 0.88\u00b10.01\u2005\u00c5, respectively, and with Uiso(H) = 1.5Ueq(O) and 1.2Ueq(N), respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989018007703/hb7752sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018007703/hb7752Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018007703/hb7752Isup3.cmlSupporting information file. DOI: 1844854CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In this compound, C\u2014H\u22efO and C\u2014H\u22efI hydrogen-bonding inter\u00adactions, \u03c0\u2013\u03c0 inter\u00adactions, other short contacts and Pb octa\u00adhedral chains are present, extending the crystal structure into a three-dimensional supra\u00admolecular network.The compound tris\u00ad bis\u00ad(dimethyl sulfoxide)di-\u03bc 3-iodido-tetra-\u03bc2-iodido-octa\u00adiodido\u00adtetra\u00adlead(II) dimethyl sulfoxide di\u00adsolvate, (C18H26N2)3[Pb4I14(C2H6OS)2]\u00b72C2H6OS, belongs to a class of organic\u2013inorganic hybrid materials with novel functionalities. In this compound, C\u2014H\u22efO and C\u2014H\u22efI hydrogen-bonding inter\u00adactions, \u03c0\u2013\u03c0 inter\u00adactions, other short contacts and Pb octa\u00adhedral chains are present, extending the crystal structure into a three-dimensional supra\u00admolecular network.The title compound, tris\u00ad bis\u00ad(dimethyl sulfoxide)di-\u03bc Viologens show excellent redox and chemical stability. In addition, they can act as effective templates for the construction of various organic\u2013inorganic hybrids, charge-transfer complexes and supra\u00admolecular systems halide-based organic\u2013inorganic hybrids possess a large radius, a flexible coordination environment, and variable stereochemical activities of the lead center ]6\u2212 trianion, one and a half BV2+ dications and two DMSO mol\u00adecules, as shown in Fig.\u00a012+ cation is located on a general position and adopts a non-planar structure, with a dihedral angle of 27.5\u2005(3)\u00b0 between the planes of the pyridinium rings. In the bipyridinium rings, C\u2014N bond lengths vary from 1.335\u2005(9) to 1.499\u2005(10)\u2005\u00c5 and C\u2014C bond lengths from 1.336\u2005(17) to 1.636\u2005(17)\u2005\u00c5. C\u2014N\u2014C bond angles are in the range 118.6\u2005(6)\u2013121.1\u2005(7)\u00b0 and C\u2014C\u2014C bond angles in the range 107.9\u2005(9)\u2013122.1\u2005(6)\u00b0. The inorganic anion can be considered as a set of mixed face-shared/edge-shared octa\u00adhedra lone pairs ]6\u2212 and DMSO via C\u2014H\u22efI and C\u2014H\u22efO hydrogen bonds (Table\u00a014I14)]6\u2212 and organic species play an important role in stabilizing the crystal structure to 4.796\u2005(4)\u2005\u00c5 (DMF)2(DPB)5] [Pb3I9] [Pb2I6] iodide complexes have been reported whose structures include chains of face-sharing ideal PbI2 and 10\u2005ml of methanol were stirred under an argon atmosphere until dissolved. 1,1\u2032-Dibutyl-4,4\u2032-bipyridyl cation salt dissolved in methanol (5\u2005ml) was added to the reaction mixture at room temperature. The resulting precipitate was dissolved in DMSO (3\u2005ml) and placed in a sealed jar of anhydrous ether. Red crystals were produced two weeks later under an argon-protected atmosphere. After filtering and drying under vacuum, red needle-shaped crystals of 0.73\u2005g (72.3%) with high quality were obtained. Analysis calculated for C62H102I14N6O4Pb4S4: C 19.97, H 2.70, N 2.25%. Found: C 19.80, H 2.82, N 2.25%. IR (cm\u22121): 3291 (w), 3108 (m), 3035 (s), 2931 (w), 2958 (w), 2857 (w), 944 (w), 1636 (m), 1634 (s), 1441 (m), 1060 (s), 833 (s).NaI , PbIUiso(H)= 1.2-1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018016584/ex2016sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018016584/ex2016Isup2.hklStructure factors: contains datablock(s) I. DOI: 1880239CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Bnz\u22efOThz (Bnz = benzene and Thz = thia\u00adzine) hydrogen bonds form corrugated chains extending along the b-axis direction which are tied into layers parallel to the bc plane by inter\u00admolecular C\u2014HMethy\u22efSThz (Methy = methyl\u00adene) hydrogen bonds, enclosing The title compound contains 1,4-benzo\u00adthia\u00adzine and 2,4-di\u00adchloro\u00adbenzyl\u00adidene units, where the di\u00adhydro\u00adthia\u00adzine ring adopts a screw-boat conformation. In the crystal, inter\u00admolecular C\u2014H 22H15Cl2NOS, contains 1,4-benzo\u00adthia\u00adzine and 2,4-di\u00ad\u00adchloro\u00adbenzyl\u00adidene units, where the di\u00adhydro\u00adthia\u00adzine ring adopts a screw-boat conformation. In the crystal, inter\u00admolecular C\u2014HBnz\u22efOThz (Bnz = benzene and Thz = thia\u00adzine) hydrogen bonds form corrugated chains extending along the b-axis direction which are connected into layers parallel to the bc plane by inter\u00admolecular C\u2014HMethy\u22efSThz (Methy = methyl\u00adene) hydrogen bonds, en\u00adclosing R44(22) ring motifs. Offset \u03c0-stacking inter\u00adactions between 2,4-di\u00ad\u00adchloro\u00adphenyl rings [centroid\u2013centroid = 3.7701\u2005(8)\u2005\u00c5] and \u03c0-inter\u00adactions which are associated by C\u2014HBnz\u22ef\u03c0(ring) and C\u2014HDchlphy\u22ef\u03c0(ring) inter\u00adactions may be effective in the stabilization of the crystal structure. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (29.1%), H\u22efC/C\u22efH (27.5%), H\u22efCl/Cl\u22efH (20.6%) and O\u22efH/H\u22efO (7.0%) inter\u00adactions. Hydrogen-bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Computational chemistry indicates that in the crystal, the C\u2014HBnz\u22efOThz and C\u2014HMethy\u22efSThz hydrogen-bond energies are 55.0 and 27.1\u2005kJ\u2005mol\u22121, respectively. Density functional theory (DFT) optimized structures at the B3LYP/6-311G level are compared with the experimentally determined mol\u00adecular structure in the solid state. The HOMO\u2013LUMO behaviour was elucidated to determine the energy gap.The title compound, C The mol\u00adecular and crystal structures, together with the Hirshfeld surface analysis, the inter\u00admolecular inter\u00adaction energies and density functional theory (DFT) computational calculations were carried out at the B3LYP/6-311G and B3LYP/6-311G levels, respectively, for (I)1,4-Benzo\u00adthia\u00adzine derivatives constitute an important class of heterocyclic systems. These mol\u00adecules exhibit a wide range of biological applications, indicating the fact that the 1,4-benzo\u00adthia\u00adzine moiety is a template potentially useful in medicinal chemistry research and therapeutic applications, such as the anti-inflammatory (Trapani B (atoms S1/N1/C1/C6\u2013C8), adopts a screw-boat conformation with puckering parameters \u2005\u00c5, \u03b8 = 68.34\u2005(16)\u00b0 and \u03c6 = 333.95\u2005(17)\u00b0. The planar rings A (C1\u2013C6), C (C10\u2013C15) and D (C17\u2013C22) are oriented at dihedral angles of A/C = 60.49\u2005(4)\u00b0, A/D = 79.69\u2005(4)\u00b0 and C/D = 41.29\u2005(4)\u00b0. Atoms Cl1 and Cl2 are \u22120.0156\u2005(3) and 0.0499\u2005(4)\u2005\u00c5 from ring C and so are almost coplanar.The title compound, (I), contains 1,4-benzo\u00adthia\u00adzine and 2,4-di\u00adchloro\u00adbenzyl\u00adidene units Fig.\u00a01, where tBnz\u22efOThz (Bnz = benzene and Thz = thia\u00adzine) hydrogen bonds form corrugated chains extending along the b-axis direction which are connected into layers parallel to the bc plane by inter\u00admolecular C\u2014HMethy\u22efSThz (Methy = methyl\u00adene) hydrogen bonds, enclosing et al., 1995C , may further stabilize the structure, with a centroid\u2013centroid distance of 3.7701\u2005(8)\u2005\u00c5, together with \u03c0-inter\u00adactions, i.e. C\u2014HBnz\u22ef\u03c0(ring) and C\u2014HDchlphy\u22ef\u03c0(ring) . The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (29.1%), H\u22efC/C\u22efH (27.5%), H\u22efCl/Cl\u22efH (20.6%) and O\u22efH/H\u22efO (7.0%) inter\u00adactions. Hydrogen-bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing.In the crystal, inter\u00admolecular C\u2014HCrystalExplorer and those delineated into H\u22efH, H\u22efC/C\u22efH, H\u22efCl/Cl\u22efH, O\u22efH/H\u22efO, C\u22efC, S\u22efH/H\u22efS and Cl\u22efC/C\u22efCl contacts \u2013(h), respectively, together with their relative contributions to the Hirshfeld surface. The most important inter\u00adaction is H\u22efH, contributing 29.1% to the overall crystal packing, which is reflected in Fig.\u00a06b) as widely scattered points of high density due to the large hydrogen content of the mol\u00adecule with the tip at de = di = 1.17\u2005\u00c5, due to the short inter\u00adatomic H\u22efH contacts (Table\u00a02c), with a 27.5% contribution to the HS, arises from the H\u22efC/C\u22efH contacts , with a 20.6% contribution to the HS, has a symmetrical distribution of points with the edges at de + di = 2.78\u2005\u00c5 arising from the H\u22efCl/Cl\u22efH contacts (Table\u00a02e), with a 7.0% contribution to the HS, arises from the O\u22efH/H\u22efO contacts have an arrow-shaped distribution of points with the tip at de = di = 1.7\u2005\u00c5. Finally, the characteristic wings resulting in the fingerprint plots delineated into S\u22efH/H\u22efS and Cl\u22efC/C\u22efCl contacts , with 4.0 and 2.2% contributions to the HS, arise from the S\u22efH/H\u22efS and Cl\u22efC/C\u22efCl contacts rm Fig.\u00a03, the whis Table\u00a02. In the s Table\u00a02c, with s Table\u00a02 and are \u22efH Fig.\u00a06d, with s Table\u00a02. The pais Table\u00a02e, with s Table\u00a02 and is vts Fig.\u00a06f have as Figs.\u00a06g and 6hs Table\u00a02 and are dnorm plotted onto the surface are shown for the H\u22efH, H\u22efC/C\u22efH, H\u22efCl/Cl\u22efH, O\u22efH/H\u22efO, C\u22efC and S\u22efH/H\u22efS inter\u00adactions in Figs.\u00a07a)\u2013(f), respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH, H\u22efCl/Cl\u22efH and O\u22efH/H\u22efO inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the biggest roles in the crystal packing is the sum of the electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) energies were calculated as \u221220.3 (Eele), \u22122.6 (Epol), \u221279.4 (Edis), 60.7 (Erep) and \u221255.0 (Etot) for C\u2014HBnz\u22efOThz hydrogen-bonding inter\u00adactions, and \u22125.8 (Eele), \u22121.0 (Epol), \u221251.0 (Edis), 39.3 (Erep) and \u221227.1 (Etot) for C\u2014HMethy\u22efSThz hydrogen-bonding inter\u00adactions.The inter\u00admolecular inter\u00adaction energies are calculated using CE\u2013B3LYP/6-31G energy model available in via density functional theory (DFT) using standard B3LYP functional and 6-311G basis-set calculations , hardness (\u03b7), potential (\u03bc), electrophilicity (\u03c9) and softness (\u03c3) are recorded in Table\u00a04E = ELUMO \u2013 EHOMO) of the mol\u00adecule was about 5.3364\u2005eV, and the frontier mol\u00adecular orbital (FMO) energies, EHOMO and ELUMO, were \u22128.2479 and \u22122.9115\u2005eV, respectively.The optimized structure of (I)et al., 2016R1 = Ph and R2 = C; see Scheme\u00a02R1 = Ph and R2 = CH2C\u2261CH (IIa) to 36\u00b0 (for IIf). The other two (IIa and IIc) have the benzo\u00adthia\u00adzine unit nearly planar, with corresponding dihedral angles of ca 3\u20134\u00b0.A search in the Cambridge Structural Database (Groom Z)-2--2H-1,4-benzo\u00adthia\u00adzin-3(4H)-one (3.21\u2005mmol), benzyl chloride (6.52\u2005mmol) and potassium carbonate (6.51\u2005mmol) in di\u00admethyl\u00adformamide was added a catalytic amount of tetra-n-butyl\u00adammonium bromide (0.33\u2005mmol). The mixture was stirred for 24\u2005h. The solid material was removed by filtration and the solvent evaporated under vacuum. The solid product was purified by recrystallization from ethanol to afford colourless crystals in 82% yield.To a solution of I, global. DOI: 10.1107/S2056989019013586/lh5925Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019013586/lh5925Isup3.cdxSupporting information file. DOI: 1957875, 1957875CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, offset \u03c0\u2013\u03c0 inter\u00adactions involving inversion-related pyridine rings [centroid-to-centroid distance = 3.4731\u2005(14)\u2005\u00c5] link the mol\u00adecules into columns along the 14H14ClNO3, there is an intra\u00admolecular C\u2014H\u22efO hydrogen bond forming an S(6) graph-set motif. The mol\u00adecule is essentially planar with the mean plane of the ethyl acetate group making a dihedral angle of 5.02\u2005(3)\u00b0 with the ethyl 6-chloro-2-eth\u00adoxy\u00adquinoline mean plane. In the crystal, offset \u03c0\u2013\u03c0 inter\u00adactions with a centroid-to-centroid distance of 3.4731\u2005(14)\u2005\u00c5 link inversion-related mol\u00adecules into columns along the c-axis direction. Hirshfeld surface analysis indicates that H\u22efH contacts make the largest contribution (50.8%) to the Hirshfeld surface.In the title quinoline derivative, C We report herein on its crystal and mol\u00adecular structures along with the Hirshfeld surface analysis.Quinoline derivatives represent an important class of bioactive heterocyclic compounds in the field of pharmaceuticals (Chu a. The mol\u00adecule consists of a quinoline fused-ring system (N1/C1\u2013C9) with meth\u00adoxy\u00adethane (O2/C10/C11), ethyl acetate (O3/O4/C13/C14) and a chlorine atom (Cl1) substituents. The intra\u00admolecular C5\u2014H5A\u22efO3 hydrogen bond (Table\u00a01S(6) graph-set motif, stabilizing the mol\u00adecular structure and preventing free rotation between the 6-chloro\u00adquinoline ring (Cl1/N1/C1\u2013C9) and the ethyl acetate (O3/O4/C12\u2013C14) moiety. Additionally, the presence of this intra\u00admolecular C\u2014H\u22efO inter\u00adaction leads to an essentially planar mol\u00adecular structure , where the ethyl acetate (O3/O4/C12\u2013C14) mean plane is twisted slightly at a dihedral angle of 5.02\u2005(3)\u00b0 with respect to the mean plane of the ethyl 6-chloro-2-eth\u00adoxy\u00adquinoline (Cl1/O2/C1\u2013C11) moiety. This essentially planar mol\u00adecular structure may be considered an important binding mode that can enhance biological activity . They are linked by offset \u03c0\u2013\u03c0 inter\u00adactions involving inversion-related pyridine rings. These inter\u00adactions link the mol\u00adecules into columns up the c-axis direction with a centroid-to-centroid (Cg\u22efCgi) distance of 3.4731\u2005(14)\u2005\u00c5 .In the crystal, mol\u00adecules lie in a plane parallel to the contact distances from the Hirshfeld surface to the nearest atom inside and outside enables the analysis of the inter\u00admolecular inter\u00adactions through the mapping of dnorm. The Hirshfeld surfaces (HS) mapped over the electrostatic potential (\u22120.0534 to 0.0319 atomic units) and dnorm (\u22120.0210 to 1.4779 arbitrary units) are shown in Fig.\u00a03a and 3b. The red spots on the Hirshfeld surface indicate inter\u00adactions involved in H\u22efO contacts. The \u03c0\u2013\u03c0 stacking is confirmed by the small blue regions surrounding bright red spots in the aromatic ring in Fig.\u00a03c, the Hirshfeld surface mapped over the shape-index, and by the flat regions around the aromatic regions in Fig.\u00a03d, the Hirshfeld surface mapped over the curvedness.The Hirshfeld surface analysis points associated with the hydrogen atoms is shown in Fig.\u00a05b. It is characterized by an end point that points to the origin, indicating the presence of the H\u22efH contacts that contribution 50.8%. The Cl\u22efH/H\u22efCl contacts between the chlorine atoms inside the Hirshfeld surface and the hydrogen atoms outside the surface and vice versa contribute 16.0% . The O\u22efH/H\u22efO (10.3%) plot shows two symmetrical wings on the left and right sides . The C\u22efC contacts contribute 7.9% , the C\u22efH/H\u22efC contacts contribute 5.3% , followed by the C\u22efO contacts at 3.7% and the C\u22efN contacts at 3.3% .There are no significant classical inter\u00admolecular contacts present in the crystal according to the analysis of the crystal structure using 0% Fig.\u00a05c. The Oes Fig.\u00a05d. The C9% Fig.\u00a05e, the C3% Fig.\u00a05e, follo7% Fig.\u00a05g and th3% Fig.\u00a03h.Gaussian 09 package was also calculated using DFT methods at the B3LYP/6-311+G level of theory using the et al., 2016et al., 2010aet al., 2010bet al., 2010cet al., 2010det al., 2009et al., 2009A search of the Cambridge Structural Database of tetra-n-butyl\u00adammonium bromide (TBAB). The reaction mixture was stirred at room temperature in DMF for 24\u2005h. After removal of salts by filtration, the DMF was evaporated under reduced pressure and the residue obtained was dissolved in di\u00adchloro\u00admethane\u00b7The organic phase was dried over Na2SO4 then concentrated in vacuo. The resulting mixture was chromatographed on a silica gel column [eluent: ethyl acetate/hexane (1:9 v/v)]. Colourless crystals were obtained when the solvent was allowed to evaporate (yield: 32%).A solution of 0.5\u2005g (1.99\u2005mmol) of ethyl 6-chloro-2-oxo-1,2-di\u00adhydro\u00adquinoline-4-carboxyl\u00adate in 25\u2005ml of DMF was mixed with 0.3\u2005ml (3.98\u2005mmol) of bromo\u00adethane, 0.55\u2005g (3.98\u2005mmol) of KUiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms. A rotating group model was applied to the methyl groups.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019007473/mw2143sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019007473/mw2143Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019007473/mw2143Isup3.cmlSupporting information file. DOI: 1890687CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E) -meth\u00adoxy\u00adbenzaldehyde oxime derivatives, namely \u22efO(hy\u00addroxy) hydrogen bonds generate C(3) chains in 1 and 2. In contrast, in compound 3, the O\u2014H(oxime)\u22efO(hy\u00addroxy) hydrogen bonds generate symmetric 4 from the O\u2014H(oxime)\u22efO(hy\u00addroxy) and C\u2014H(2-meth\u00adoxy)\u22efO(hy\u00addroxy) hydrogen bonds.The crystal structures of four ( E)-meth\u00adoxy\u00adbenzaldehyde oxime derivatives, namely \u22efO(hy\u00addroxy) hydrogen bonds generate C(3) chains in 1 and 2. In contrast, in compound 3, the O\u2014H(oxime)\u22efO(hy\u00addroxy) hydrogen bonds generate symmetric R22(6) dimers. A more complex dimer is generated in 4 from the O\u2014H(oxime)\u22efO(hy\u00addroxy) and C\u2014H(2-meth\u00adoxy)\u22efO(hy\u00addroxy) hydrogen bonds. In all cases, further inter\u00adactions, C\u2014H\u22efO and C\u2014H\u22ef\u03c0 or \u03c0\u2013\u03c0, generate three-dimensional arrays. Hirshfeld surface and fingerprint analyses are discussed.The crystal structures of four ( The last survey of the classical hydrogen-bonding patterns in benzaldehyde oximes reported in 2010 . In compounds 1\u20133, the 2-meth\u00adoxy group and the hydrogen of the oxime moiety have an s-cis arrangement. In contrast, in both mol\u00adecules of compound 4, the 2-meth\u00adoxy group and the hydrogen atom of the oxime moiety have an s-trans arrangement. The s-trans arrangement of the 2-alk\u00adoxy group and hydrogen atom of the oxime units in compound 4 is very much rarer than the s-cis arrangement found in compounds 1\u20133 and other non-salicylaldoximes. A search of the Cambridge Structural Database -2-({2-[(E)-(hy\u00addroxy\u00adimino)\u00admeth\u00adyl]phen\u00adoxy}meth\u00adyl)-3-p-tolyl\u00adacryl\u00ado\u00adnitrile meth\u00adyl]phen\u00adoxy}meth\u00adyl)-3-(2-methyl\u00adphen\u00adyl)acrylo\u00adnitrile -2-({2-[(E)-(hy\u00addroxy\u00adimino)\u00admeth\u00adyl]phen\u00adoxy}meth\u00adyl)-3-phenyl\u00adacrylo\u00adnitrile chains, illustrated in Fig.\u00a061 are two weaker hydrogen bonds, namely, C3\u2014H3\u22efO13ii and C21\u2014H21C\u22efO13iii, as well as a weak \u03c0\u2013\u03c0 stacking inter\u00adaction . These three inter\u00adactions generate the mol\u00adecular arrangement shown in Fig.\u00a07ii hydrogen bonds generate C7 chains in the c-axis direction, while the C21\u2014H21C\u22efO13iii hydrogen bonds form C(8) spiral chains along the a-axis direction: together these hydrogen bonds form c-axis direction. The involvement of the weaker C3\u2014H3\u22efO13ii, C21\u2014H21C\u22efO13iii and \u03c0\u2013\u03c0 inter\u00adactions, along with the stronger O13\u2014H13 \u22efN12i hydrogen bonds, creates the three-dimensional structure for 1.In the crystal of s Table\u00a02, forming1, mol\u00adecules of 2 are primarily linked by strong O13\u2014H13 \u22efN12i hydrogen bonds (Table\u00a03C(3) chains: as such chains are very similar to those in compound 1, see Fig.\u00a06C(3) chain in compound 2. Other inter\u00admolecular inter\u00adactions in 2 are the weaker C21\u2014H21B\u22efO31iii and C31\u2014H31B\u22efO13iv hydrogen bonds and a C31\u2014H31C\u22efCg1v inter\u00adaction involving the C1\u2013C6 ring. These three inter\u00adactions combine to form the arrangement illustrated in Fig.\u00a08B\u22efO31iii hydrogen bonds on their own generate C(6) chains, which propagate in the a-axis direction while the C31\u2014H31B\u22efO13iv hydrogen bonds generate spiral C(9) chains in the b-axis direction. Together these hydrogen bonds generate a network of C\u22efCg1v inter\u00adactions lead to chains along the a-axis direction. The involvement of the weaker C21\u2014H21B\u22efO31iii, C31\u2014H31B\u22efO13iv C and C\u2014H\u22ef\u03c0 inter\u00adactions, along with the stronger O13\u2014H13 \u22efN12i hydrogen bonds, creates a three-dimensional structure for 2. C4\u2014H4\u22efO12ii hydrogen bonds also occur.As in s Table\u00a03, forming3, i hydrogen bonds (Table\u00a04A(meth\u00adoxy)\u22efO13ii hydrogen bonds provide a two-mol\u00adecule-wide ribbon. Within the ribbons are 3 is the C41\u2014H41C\u22efCg1iii inter\u00adaction, which generates a tilted ladder assembly, propagating in the a-axis direction, with the C\u22efCg1iii inter\u00adactions as the supports.In compound s Table\u00a04, as illu4, each of the two independent mol\u00adecules forms symmetric dimers, see Fig.\u00a010i and O113\u2014H113\u22efO121i hydrogen bonds chains, while in compounds 3 and 4, they are responsible for the creation of the dimers. In compound 3, the fins ending at de, di = 1.9,1.1\u2005\u00c5 are due to C(\u03c0)\u22efH/C(\u03c0)\u22efH contacts. The FP plots for Mol A and Mol B of compound 4 are asymmetric because of the different inter\u00adactions of each mol\u00adecule. The double wings in the FP plot for Mol A in the second quadrant are complementary to those displayed in the fourth quadrant by MolB and relate to C\u22efH close contacts connecting the two mol\u00adecules. The spike ending at di, de = 1.1\u2005\u00c5 in Mol A is due to H\u22efH contacts.Hirshfeld surfaces -chain-forming compounds 1 and 2 show higher percentages of H\u22efH and C\u22efC contacts, but a lower percentage of H\u22efC/C\u22efH contacts, than the dimer-forming compounds 3 and 4.The percentages of the various atom\u2013atom contacts, derived from the fingerprint plots, for the four compounds are shown in Table\u00a06et al., 20162 and 3. The classical hydrogen bonds in 3,5-di\u00admeth\u00adoxy\u00adbenzene oxime generate C(3) chains in the CSD database with oxime 1, 371\u2013373\u2005K for 2, 378\u2013380\u2005K for 3 and 370\u2013371\u2005K for 4.The title compounds were prepared from hy\u00addroxy\u00adamine and the corresponding benzaldehyde in methanol in the presence of potassium carbonate and were recrystallized from methanol solutions, m.p. = 364\u2013365\u2005K for compound Uiso(H) = 1.2\u20131.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0710.1107/S2056989018014020/qm2129sup1.cifCrystal structure: contains datablock(s) 1, 2, 3, 4, global. DOI: 10.1107/S2056989018014020/qm21291sup2.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989018014020/qm21292sup3.hklStructure factors: contains datablock(s) 2. DOI: 10.1107/S2056989018014020/qm21293sup4.hklStructure factors: contains datablock(s) 3. DOI: 10.1107/S2056989018014020/qm21294sup5.hklStructure factors: contains datablock(s) 4. DOI: Click here for additional data file.10.1107/S2056989018014020/qm21291sup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018014020/qm21292sup7.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018014020/qm21293sup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018014020/qm21294sup9.cmlSupporting information file. DOI: 1871165, 1871164, 1871163, 1871162CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The supra\u00admolecular structure is mainly governed by C\u2014H\u22efN hydrogen-bonded centrosymmetric dimers, C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds and S\u22ef\u03c0 stacking inter\u00adactions, which together lead to the formation of a layered crystal packing.The mol\u00adecular conformation is stabilized 15H13N3O2S, crystallizes in the monoclinic space group P21/n and its mol\u00adecular conformation is stabilized via intra\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN contacts. The supra\u00admolecular structure is mainly governed by C\u2014H\u22efN hydrogen-bonded centrosymmetric dimers, C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds and S\u22ef\u03c0 and \u03c0\u2013\u03c0 stacking inter\u00adactions which, together, lead to the formation of a layered crystal packing. The inter\u00admolecular inter\u00adactions were further evaluated through the mol\u00adecular electrostatic potential map and Hirshfeld fingerprint analysis.The title compound, C N-heterocycles (having two carbon and three nitro\u00adgen atoms) because of their unique structural and chemical properties methanone (1).In general, the 1,2,3-triazole nucleus is the most fundamental heterocyclic component found in various pharmacologically active agents in the asymmetric unit \u2005\u00c5] and C11\u2014H11\u22efN1 [C11\u22efN1 = 2.950\u2005(2)\u2005\u00c5] contacts of the meth\u00adoxy group with the triazol nitro\u00adgen N3 [C15\u22efN3 = 3.490\u2005(3)\u2005\u00c5] and the thio\u00adphene hydrogen H12 (sp2) with the triazol nitro\u00adgen N1 [C12\u22efN1 = 3.768\u2005(2)\u2005\u00c5]. These are extended in an alternate fashion, forming ribbons along the [101] direction [3.492\u2005(2)\u2005\u00c5] inter\u00adactions along the [010] direction, forming a corrugated sheet perpendicular to the (101) plane , and two-dimensional fingerprint plots (thio\u00adphen-2-yl)methanone subunit resulted in one hit =1.5Ueq(C) for the methyl group and C\u2014H = 0.95\u00c5 with Uiso(H) = 1.2Ueq(C) for the aromatic C atoms.Crystal data, data collection and structure refinement details are given in Table\u00a0210.1107/S2056989018010654/xi2009sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018010654/xi2009Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018010654/xi2009Isup3.cmlSupporting information file. DOI: 1850683CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "While there are two C\u2014H\u22efO weak inter\u00admolecular inter\u00adactions present in both (I) and (II), the change of substituent from chlorine to methyl has given rise to an additional weak C\u2014H\u22efO inter\u00admolecular inter\u00adaction that is relatively stronger than the other two. However, the presence of the stronger C\u2014H\u22efO inter\u00adaction in (II) has not disrupted the validity of the chloro-methyl exchange rule. Details of the crystal structures and Hirshfeld analyses of the two compounds are presented.Instances of crystal structures that remain isomorphous in spite of some minor changes in their respective mol\u00adecules, such as change in a substituent atom/group, can provide insights into the factors that govern crystal packing. In this context, an accurate description of the crystal structures of an isomorphous pair that differ from each other only by a chlorine\u2013methyl substituent, The prediction of crystal structures has emerged as an exciting field involving researchers from diverse fields primarily because of its challenging complexity, which is considered analogous to that of the protein-folding problem. Attempts made in the field of crystal-structure prediction, its present status and the challenges ahead were discussed in detail in a recent article et al., 2011et al., 2018et al., 2019et al., 2007et al., 2008From a pharmacological view point, the title compounds (I)The mol\u00adecular structures of (I)Q(2) = 0.4011\u2005(2)\u2005\u00c5 and \u03c6 = 180.3733\u2005(3)\u00b0 for (I)Q(2) = 0.4047\u2005(2)\u2005\u00c5 and \u03c6 = 180.3444\u2005(3)\u00b0 for (II)Q = 0.5572\u2005(16)\u2005\u00c5, \u03b8 = 138.9\u2005(2)\u00b0, \u03c6 = 219.8\u2005(3)\u00b0 in (I)Q = 0.5603\u2005(17)\u2005\u00c5, \u03b8 = 137.7\u2005(2)\u00b0, \u03c6 = 219.6\u2005(3)\u00b0 in (II)As expected, the conformational features of both compounds are nearly identical, as shown in a overlay diagram Fig.\u00a03. The fivThe dihedral angle between the mean planes of the two chloro\u00adphenyl groups in (I)viz. C10\u2014H10\u22efO2 and C16\u2014H16\u22efO2, which are identical in nature and characteristic of similar fundamental mol\u00adecular inter\u00adaction patterns are present (Tables 3b-axis direction in both (I)b-axis direction Tables 3 and 4 \u25b8.viz. C36\u2014H36A\u22efO1, that is stronger than the two characteristic weak inter\u00admolecular inter\u00adactions and involves the replaced substituent methyl group (C36\u2014H36A) as a donor and the piperidinone O1 atom as an acceptor (see Table\u00a04In (II)B\u22ef\u03c0 inter\u00adaction observed in (I)Cg3\u22efCg3 distances of 3.7459\u2005(2)\u2005\u00c5 in (I)Cg3 is the centroid of the C14\u2013C19 ring. The shortest Cl\u22efCl distance observed [Cl1 \u22efCl1] is 4.088\u2005(1)\u2005\u00c5 and bears no structural significance.In addition, a weak C\u2014H\u22ef\u03c0 inter\u00adaction involving different donor groups and acceptor \u03c0-ring systems is present in both (I)et al., 2016R factors less than 0.05) gave only three hits: 5\u2032\u2032-(4-chloro\u00adbenzyl\u00adidene)-4\u2032-(4-chloro\u00adphen\u00adyl)-1\u2032,1\u2032\u2032-dimethyl-2H,4\u2032\u2032H-di\u00adspiro\u00ad-2,4\u2032\u2032-dione -pyridinone analysis was used to investigate and visualize the weak inter\u00admolecular inter\u00adactions influential in the packing of the mol\u00adecules in the crystal. The visual representation of mol\u00adecular inter\u00adactions on this isosurface is determined using two parameters, CrystalExplorer3.0 E)-2-chloro\u00adphenyl\u00admethyl\u00adidene] tetra\u00adhydro-4(1H)- pyridinone (1\u2005mmol), acenaphthene\u00adquinone (1\u2005mmol) and sarcosine (1\u2005mmol) was dissolved in methanol (15\u2005mL) and refluxed for 30\u2005min. After completion of the reaction, as evident from TLC, the mixture was poured into water (50\u2005mL) and the precipitated solid was filtered and washed with water (100\u2005mL) to obtain pure (I)fR 0.40. Suitable crystals for single-crystal X-ray studies were obtained by recrystallization of the product from ethanol.For (I)E)-2-methyl\u00adphenyl\u00admethyl\u00adidene] tetra\u00adhydro-4(1H)-pyridinone (1\u2005mmol), acenaphthene\u00adquinone (1\u2005mmol) and sarcosine (1\u2005mmol) in methanol (15\u2005mL) to yield yellow crystals.A similar procedure for (II)Uiso(H) = 1.5Ueq(C) for methyl H atoms or 1.2Ueq(C) otherwise. The H atoms of the methyl atoms C35 and C36 in (II)Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989019012428/jj2215sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989019012428/jj2215Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019012428/jj2215IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1951894, 1569029CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II ion is coordinated by two N atoms of the N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adethylenedi\u00adamine ligand and two S atoms from two 2-meth\u00adoxy\u00adethyl xanthate ligands. Two C\u2014H\u22efO and two C\u2014H\u22efS intra\u00admolecular inter\u00adactions occur. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds, forming a three-dimensional supra\u00admolecular architecture.In the title compound, the Zn S)zinc(II) acetone hemisolvate, [Zn(C4H7O2S2)2(C6H16N2)]\u00b70.5C3H6O, the ZnII ion is coordinated by two N atoms of the N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adethylenedi\u00adamine ligand and two S atoms from two 2-meth\u00adoxy\u00adethyl xanthate ligands. The amine ligand is disordered over two orientations and was modelled with refined occupancies of 0.538\u2005(6) and 0.462\u2005(6). The mol\u00adecular structure features two C\u2014H\u22efO and two C\u2014H\u22efS intra\u00admolecular inter\u00adactions. In the crystal, mol\u00adecules are linked by weak C\u2014H\u22efO and C\u2014H\u22efS hydrogen bonds, forming a three-dimensional supra\u00admolecular architecture. The mol\u00adecular structure was optimized using density functional theory (DFT) at the B3LYP/6\u2013311\u2005G level. The smallest HOMO\u2013LUMO energy gap (3.19\u2005eV) indicates the suitability of this crystal for optoelectronic applications. The mol\u00adecular electrostatic potential (MEP) further identifies the positive, negative and neutral electrostatic potential regions of the mol\u00adecules. Half a mol\u00adecule of disordered acetone was removed with the solvent-mask procedure in OLEX2 and this contribition is included in the formula.In the title compound, bis\u00ad(2-meth\u00adoxy\u00adethyl xanthato-\u03ba al. 2009. J. Appl Xanthate is a bidentate monoanionic sulfur\u2013sulfur donor ligand. It stabilizes complexes of most of the transition elements and can bind metal centers in monodentate, isobidenate, anisobidenate or ionic modes. Xanthates have the ability to inhibit the replication of both RNA and DNA viruses II ion is coordinated by two N atoms of the N,N,N\u2032,N\u2032-tetra\u00admethyl\u00adethylenedi\u00adamine mol\u00adecule and two S atoms from two 2-meth\u00adoxy\u00adethylxanthate mol\u00adecules. The Zn1\u2014N1, Zn1\u2014N2, Zn1\u2014S1 and Zn1\u2014S3 bond lengths are 2.141\u2005(5), 2.123\u2005(5), 2.3107\u2005(9) and 2.3050\u2005(9)\u2005\u00c5, respectively \u2005\u00c5, which is typical of a single bond whereas the carbon-to-sulfur S2 distance of 1.647\u2005(3)\u2005\u00c5 is typical of a carbon-to-sulfur double bond. In the mol\u00adecule, weak C1\u2014H1C\u22efO8, C2A\u2014H2AB\u22efO11, C5A\u2014H5AA\u22efS1 and C6\u2014H6C\u22efS4 intra\u00admolecular inter\u00adactions are observed 2(bipy)] [CSD 2(C6H16N2)] and [Ni(moexa)2phen] (benzene solvate), moexa = O-methoxy\u00adethyl\u00adxan\u00adthato-S,S\u2032 2] and the lowest unoccupied mol\u00adecular orbitals (LUMOs) are named as frontier mol\u00adecular orbitals (FMOs). The FMOs play an important role in the optical and electric properties. The frontier orbital gap characterizes the chemical reactivity and the kinetic stability of the mol\u00adecule. A mol\u00adecule with a small frontier orbital gap is generally associated with a high chemical reactivity, low kinetic stability and is also termed a soft mol\u00adecule. The density functional theory (DFT) quantum-chemical calculations for the title compound were performed at the B3LYP/6\u2013311\u2005G level level of theory and is illustrated in Fig.\u00a043CO2)\u00b72H2O in 2-meth\u00adoxy\u00adethanol. A hot solution of potassium 2-meth\u00adoxy\u00adethylxanthate in 2-meth\u00adoxy\u00adethanol was added and the mixture was stirred for 30\u2005min. Water was added to the mixture and a white precipitate was formed. The product was recrystallized from acetone.Tetra\u00admethyl\u00adethylenedi\u00adamine was added to a hot solution of Zn(CHUiso(H) = 1.5Ueq(C) for methyl H atoms and 1.2Ueq(C) otherwise. All atoms of the amine ligand are disordered and were modelled as two orientations with relative occupancies of 0.538\u2005(6) and 0.462\u2005(6). The diffuse electron density of half an acetone solvent mol\u00adecule was removed with the solvent-mask procedure implemented in OLEX2 I. DOI: 10.1107/S2056989019013148/lh5921Isup2.hklStructure factors: contains datablock(s) I. DOI: 1420207CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The difference was ascribed to the HOMO of the amide group, which expands in the vertical direction (\u03c0O) rather than in the plane (nO). S\u00b7\u00b7\u00b7X interactions in four model proteins, phospholipase A2 (PLA2), ribonuclease A (RNase A), insulin, and lysozyme, have also been analyzed. The results suggested that S\u00b7\u00b7\u00b7X interactions would be important factors that control not only the three-dimensional structure of proteins but also their functions to some extent. Thus, S\u00b7\u00b7\u00b7X interactions will be useful tools for protein engineering and the ligand design.In organic molecules a divalent sulfur atom sometimes adopts weak coordination to a proximate heteroatom (X). Such hypervalent nonbonded S\u00b7\u00b7\u00b7X interactions can control the molecular structure and chemical reactivity of organic molecules, as well as their assembly and packing in the solid state. In the last decade, similar hypervalent interactions have been demonstrated by statistical database analysis to be present in protein structures. In this review, weak interactions between a divalent sulfur atom and an oxygen or nitrogen atom in proteins are highlighted with several examples. S\u00b7\u00b7\u00b7O interactions in proteins showed obviously different structural features from those in organic molecules ( Nagao ,33 reporives (3) , bis[2-(fide (4) , \u03b2-hydro (4) [5) , and thi [5) [6) .Weak nonbonded interactions are important physicochemical forces that control the structure of proteins . Ionic iet al. [The SSC and CSC groups involved in cystine and methionine residues, however, were usually considered just as hydrophobic moieties in folded protein structures until recently, except for S\u00b7\u00b7\u00b7C(\u03c0) interactions and weaket al. suggesteet al. ,28,51,52et al. ,47. Meanet al. , but the et al. [ etc.) atomic contacts involved in 604 high-resolution (\u22642.0 \u00c5) heterogeneous X-ray structures selected from the Protein Data Bank [d = rS\u00b7\u00b7\u00b7X \u2212 vdwS \u2212 vdwX), the directionality around the S and X atoms, and the location along the amino acid sequence revealed distinct structural features of the S\u00b7\u00b7\u00b7X interactions. In case of the most frequent S\u00b7\u00b7\u00b7O interactions, both SSC and CSC S atoms tend to approach a main-chain O atom perpendicularly to the amide plane (the \u03c0O direction), and the O atom tends to approach the S atom from the backside of the S\u2013S or S\u2013C covalent bonds (the \u03c3S* direction). Similar directionalities of the S(CSC)\u00b7\u00b7\u00b7O interactions were also reported by Pal and Chakrabarti [O orbital, not \u03c0O, is usually used to form the interaction.Nonbonded S\u00b7\u00b7\u00b7X interactions in proteins have recently been pursued by several research groups ,23,18,21 et al. thoroughata Bank . Statistkrabarti . The strd \u2264 0.0 \u01fa), a probability of S\u00b7\u00b7\u00b7O contacts increases significantly, suggesting the presence of specific S\u00b7\u00b7\u00b7O interactions in proteins. According to the statistical analysis for the obtained data, four types of nonbonded S\u00b7\u00b7\u00b7X interactions have been clearly characterized, i.e., S\u2013S\u00b7\u00b7\u00b7O=C, C\u2013S\u00b7\u00b7\u00b7O=C, C\u2013S\u00b7\u00b7\u00b7N, and S\u2013S\u00b7\u00b7\u00b7S\u2013S interactions [ractions .O \u2192 \u03c3S* orbital interaction is obvious. The S\u2013S\u00b7\u00b7\u00b7O=C interactions are most frequently observed in helices, suggesting that the S\u00b7\u00b7\u00b7O interactions would support the stability. The C\u2013S\u00b7\u00b7\u00b7O=C interaction formed between a methionine side-chain and a main-chain peptide group has a similar character to the S\u2013S\u00b7\u00b7\u00b7O=C interaction, but the strength of the interaction is weaker with attenuated directionality. Similarly, the C\u2013S\u00b7\u00b7\u00b7N interaction between a methionine side-chain and a main-chain peptide group can also be characterized by the linear C\u2013S\u00b7\u00b7\u00b7N atomic alignment and the vertical access of the S atom to the amide plane, suggesting a contribution from the \u03c0N \u2192 \u03c3S* orbital interaction. On the other hand, most of close S\u00b7\u00b7\u00b7S contacts in proteins can be assigned to S\u2013S\u00b7\u00b7\u00b7S\u2013S interaction, which would be stabilized by the nS \u2192 \u03c3S* orbital interaction in a similar manner to that observed in organic crystals [A majority of close S\u00b7\u00b7\u00b7O contacts for an SSC group in proteins is assigned to S\u2013S\u00b7\u00b7\u00b7O=C interactions, which can be characterized by the linear S\u2013S\u00b7\u00b7\u00b7O atomic alignment and the vertical access of the S atom to the carbonyl plane as shown in crystals .Ab initio calculation was carried out for the model complexes (CH3SSCH3 + CH3CONHCH3 and CH3SCH3 + CH3CONHCH3) to investigate the nature of the S\u00b7\u00b7\u00b7X interactions observed in proteins [O \u2192 \u03c3S* orbital interaction lying in the carbonyl plane rather than the \u03c0 orbital expanding perpendicular to the plane. The reason for the observed discrepancy in the directionality between the S\u00b7\u00b7\u00b7O interactions in proteins and organic molecules can be explained on the basis of the HOMO levels of various carbonyl compounds . As grapO level of amide CH3CONHCH3 is remarkably raised compared with other carbonyl compounds, while the nO level remains almost unchanged. The elevation of the \u03c0O orbital would be due to the conjugation between the N lone pair and the carbonyl group. Thus, inversion of the energy levels of nO and \u03c0O would be responsible for the observed directional preferences of the S\u00b7\u00b7\u00b7O interactions in proteins.However, the \u03c0 et al. [N-acetylglucosamine-thiazoline and \u03b2-hexosaminidase and between benzophenone and porcine odorant-binding protein [\u2212 ligand and the imidazole ring of His109 in lactoperoxidase [The importance of S\u00b7\u00b7\u00b7O interactions in the enzymatic function of proteins has been pointed out for some particular cases. Taylor and Markham suggeste et al. reported protein . Importa protein . More reroxidase and betwroxidase were sugAccording to the comparison between the results from database analyses and ab initio calculation, it was clear that the directional preferences of the S\u00b7\u00b7\u00b7O interactions observed in protein structures are in accord with the profiles of the potential surfaces calculated for the isolated model complexes . An examProtein structures are generally considered to be flexible because they are governed only by weak nonbonded interactions. Therefore, coincidence of the statistical conformational preference for the interaction of proteins with the potential surface calculated for the isolated model is to be noticed and would have important implications in protein architecture. The detailed analysis revealed the following features . The linThe directionality, however, is easily affected by crystal packing force for organic molecules. Thus, the order of the factors that control molecular structure of organic molecules and proteins in the solid state can be summarized as shown in S* direction), irrespective of the types of carbonyl groups. On the other hand, the S atom tends to approach the O atom either within the carbonyl plane (in the nO direction) or from the vertical direction (in the \u03c0O direction). In the case of S\u00b7\u00b7\u00b7O(amide) interactions, the vertical direction is significantly preferred, due probably to elevation of the \u03c0O orbital. The linearity of the S\u00b7\u00b7\u00b7O interactions in organic molecules would overcome the crystal packing force, whereas the vertical nature of the S\u00b7\u00b7\u00b7O(amide) interactions may be affected by the packing force. The verticality, however, would survive in protein structures. These structural features will be informative for protein engineering and molecular design of functional organic sulfur compounds.The O atom has strong tendency to approach the S atom from the backside of the S\u2013C or S\u2013S bond (in the \u03c32 (PLA2) was the first example of such proteins [2 family. The results are summarized in this section.With a success in characterization of S\u00b7\u00b7\u00b7X interactions in proteins, we subsequently sought out particular protein families or domains, for which specific S\u00b7\u00b7\u00b7X interactions are commonly present in a wide range of the structures registered in the Protein Data Bank . Phosphoproteins . We have2 [2 family; PLA2 and snake PLA2 (sPLA2). Comprehensive search for close S\u00b7\u00b7\u00b7X contacts in the structures of the PLA2 domain, which were retrieved from protein data bank, revealed the presence of four common S\u00b7\u00b7\u00b7O interactions, i.e., S(C44)\u00b7\u00b7\u00b7O(D40), S(C61)\u00b7\u00b7\u00b7O(A55), S(C84)\u00b7\u00b7\u00b7O(C96), and S(C98)\u00b7\u00b7\u00b7O(F94), and one common S\u00b7\u00b7\u00b7N interaction, i.e., S(M8)\u00b7\u00b7\u00b7N(R100), as shown in 2 is decreased in the M8,20L mutant [PLA2 ,63, a smL mutant . This wo2 domain. For this domain group, the phylogenetic dendrogram was already analyzed by Ohno et al. [2 involved in venom of snakes inhabiting the southern islands of Japan. Mapping the common S\u00b7\u00b7\u00b7O and S\u00b7\u00b7\u00b7N interactions observed by the database analysis, we found that most of the S\u00b7\u00b7\u00b7X interactions make clusters on the dendrogram. The results suggested a possible role of S\u00b7\u00b7\u00b7X interactions in molecular evolution of proteins.On the other hand, an evolutional aspect of the S\u00b7\u00b7\u00b7X interactions was analyzed for the sPLAo et al. using th i.e., S(C26)\u00b7\u00b7\u00b7O\u03b3(T99) and S(C65)\u00b7\u00b7\u00b7O(Q69), and one common S\u00b7\u00b7\u00b7N interaction, i.e., S(C58)\u00b7\u00b7\u00b7N(P117), were characterized. The locations along the amino acid sequence are shown in RNase A is a typC65)\u00b7\u00b7\u00b7OQ, and oneIn some RNase A structures complexed with a substrate, close S\u00b7\u00b7\u00b7X contacts between the S atom of the C65 residue and the substrate were found. Examples are S(C65)\u00b7\u00b7\u00b7O(ADT) in 8RSA , S(C65)\u00b7i.e., S(C20)\u00b7\u00b7\u00b7O(E17) and S(C19)\u00b7\u00b7\u00b7O(L15), were found within A and B chains, respectively, among the 23 high-resolution (\u22642.0 \u00c5) structures. Insulin is a pepi.e., S(C127)\u00b7\u00b7\u00b7O(I124), and three common S\u00b7\u00b7\u00b7N interactions, i.e., S(C30)\u00b7\u00b7\u00b7N\u03b5(W123), S(C80)\u00b7\u00b7\u00b7N(N65), and S(C127)\u00b7\u00b7\u00b7N\u03b7(R5), were characterized as shown in Lysozyme consists2 and sPLA2 were described in reference [Details of the database analysis of the S\u00b7\u00b7\u00b7X interactions in PLAeference . SimilarThe three-dimensional structure, hence the function, of a protein is controlled by the interplay of a number of weak nonbonded interactions, such as hydrogen bond, van der Waals forces, and hydrophobic interaction. According to the results from the database analyses and theoretical calculation summarized in this review, it would be concluded that hypervalent S\u00b7\u00b7\u00b7X interactions are also a member of such weak interactions. ab initio calculation. For four particular proteins, i.e., PLA2, RNase A, insulin, and lysozyme, unique S\u00b7\u00b7\u00b7X interactions have been characterized, and some were suggested to play roles in the stability of the native structures and the functions to some extent.Sulfur-containing functional groups of cystine (an SSC group) and methionine (a CSC group) were previously considered to be just hydrophobic moieties in protein structures, but they are indeed able to form specific nonbonded interactions with nearby polar non-hydrogen atoms in folded proteins. A unique directionality of the S\u00b7\u00b7\u00b7X interactions see would bei.e., integration of the database analyses and theoretical calculation, will be useful for characterization of other weak nonbonded interactions hidden in molecules as well as protein structures. Finally, the statistical analyses using the both protein and organic molecule structure databases demonstrated that the order of the strength of the factors that control molecular structures in the solid state can be expressed as shown in"} +{"text": "The resulting compounds, bis\u00ad di\u00adsulfide, C8H6O2S4S2, and bis\u00ad diselenide, C8H6O2Se6, are isotypic.By 8H6O2S6 and C8H6O2S4Se2, are isotypic with very similar cell parameters. The complete mol\u00adecules constitute the asymmetric units, despite being chemically perfectly symmetric. The most prominant differences in the metrical parameters arise from the distinct sizes of sulfur and selenium in the dichalcogenide bridges, with C\u2014S\u2014S\u2014C and C\u2014Se\u2014Se\u2014C torsion angles of 70.70\u2005(5) and 68.88\u2005(3)\u00b0, respectively. The crystal packing is determined by weak non-classical hydrogen-bonding inter\u00adactions. One carbonyl oxygen but not the other participates in C\u2014H\u22efO inter\u00adactions zigzagging along the b axis, forming infinite chains. This is complemented by an intra\u00admolecular C\u2014H\u22efS inter\u00adaction and further inter\u00admolecular C\u2014H\u22efS (C\u2014H\u22efSe) inter\u00adactions, resulting in a three-dimensional network. The inter\u00adactions involving the bridging chalcogenides form chains protruding along the c axis.The two title compounds, C In related di\u00adsulfides they range from 52.08 to 109.82\u00b0 are essentially planar, with maximum deviations from the least-squares plane of 0.028 and 0.022\u2005\u00c5 for the di\u00adsulfide and for the diselenide, respectively, corresponding to the distances from atom S1 to the O1\u2014S1\u2014S2\u2014C1\u2014C2\u2014C3 plane in both cases. The dihedral angles between the O1\u2014S1\u2014S2\u2014C1\u2014C2\u2014C3 and the O2\u2014S5\u2014S6\u2014C6\u2014C7\u2014C8 planes are 33.8\u2005(2)\u00b0 for the di\u00adsulfide and 28.89\u2005(11)\u00b0 for the diselenide. Here, a smaller torsion angle around the dichalcogenide bridge is accompanied by a smaller angle between the two planes of the 1,3-ene-di\u00adthiol-2-one moieties.B\u22efO1i inter\u00adactions zigzagging along the b axis, forming infinite chains \u2005\u00c5 for the disufide and 3.369\u2005(5)\u2005\u00c5 for the diselenide. This is complemented by two intra\u00admolecular inter\u00adactions between the two chalcogens of the dichalcogenide bridges and the adjacent methyl substituents (C4\u2014H4A\u22efS3/Se1 and C5\u2014H5A\u22efS4/Se2) with D\u22efA distances of 3.244\u2005(2) for S3, of 3.234\u2005(2) for S4, of 3.354\u2005(4) for Se1, and of 3.341\u2005(4) for Se2. Further inter\u00admolecular C\u2014H\u22efS and C\u2014H\u22efSe inter\u00adactions contribute to the formation of a three-dimensional network. The inter\u00adactions involving the bridging chalcogenides form chains protruding along the c axis . The first such thione crystal structure was reported in 1999 by Cerrada et al., which comprises an S\u2014S-linked [C3S5\u2014C3S5]2\u2212 dianion 2Cp complexes by coordination of thiol\u00adate sulfur to iron by Kumar et al. selenolothio\u00adphen-6-yl}diselenide was formed unexpectedly by the reaction of 2-[(tri\u00adphenyl\u00adphospho\u00adnio)meth\u00adyl] thio\u00adphene chloride with sodium hydrogen selenite , and LDA was added dropwise over 5\u2005min. The mixture was allowed to stand for 35\u2005min, warmed to ice-bath temperature and after a further 10 minutes quenched with a saturated aqueous solution of NH4Cl (around 20\u2005ml). The organic phase was diluted with EtOAc, separated and the aqueous phase re-extracted with Et2O (2 \u00d7 15\u2005ml). The combined organic phases were washed with brine, dried and the solvent evaporated in vacuo to give a yellowish oil as crude product. This was purified by chromatography (silica gel), eluting with EtOAc/petroleum ether (40/60) 3:97 v/v to give 4-methyl-5-tri-n-butyl\u00adstannyl-1,3-di\u00adthiol-2-one as the major product. During purification, a yellowish oily fraction was isolated and subsequently stored at 253\u2005K, forming large yellow crystals. Crystallographic evaluation of these crystals reveals the formation of the side product bis\u00ad di\u00adsulfide.Preparation of bis\u00ad diselenide: The synthesis was carried out under an inert gas atmosphere of nitro\u00adgen, whereas the purification steps were carried out in air. To a solution of 4-methyl-5-tri-n-butyl\u00adstannyl-1,3-di\u00adthiol-2-one in freshly distilled dioxane (5\u2005ml) was added freshly sublimed selenium dioxide . The reaction mixture was heated at reflux temperature for 6\u2005h. After cooling, the solution was filtered through celite. Solvent removal gave an orange solid . Yellow crystals suitable for crystallographic analysis were obtained by recrystallization from acetone.iUso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018007454/wm5446sup1.cifCrystal structure: contains datablock(s) CSV72a12, it14ii. DOI: 10.1107/S2056989018007454/wm5446CSV72a12sup2.hklStructure factors: contains datablock(s) CSV72a12. DOI: 10.1107/S2056989018007454/wm5446it14iisup3.hklStructure factors: contains datablock(s) it14ii. DOI: 1843766, 1843765CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the 1:1 cocrystal of nicotinamide and 2-chloro-5-nitro\u00adbenzoic acid, the mol\u00adecules form hydrogen bonds through O\u2014H\u22efN, N\u2014H\u22efO, and C\u2014H\u22efO inter\u00adactions along with N\u2014H\u22efO dimer hydrogen bonds of nicotinamide. Further additional weak \u03c0\u2013\u03c0 inter\u00adactions stabilize the mol\u00adecular assembly of this cocrystal. 7H4ClNO4\u00b7C6H6N2O, nicotinamide (NIC) and 2-chloro-5-nitro\u00adbenzoic acid (CNBA) cocrystallize with one mol\u00adecule each of NIC and CNBA in the asymmetric unit. In this structure, CNBA and NIC form hydrogen bonds through O\u2014H\u22efN, N\u2014H\u22efO and C\u2014H\u22efO inter\u00adactions along with N\u2014H\u22efO dimer hydrogen bonds of NIC. Further additional weak \u03c0\u2013\u03c0 inter\u00adactions stabilize the mol\u00adecular assembly of this cocrystal.In the title 1:1 cocrystal, C In the asymmetric unit, an (CNBA)O\u2013H\u22efN inter\u00adaction plays a prime role in the mol\u00adecular recognition of this cocrystal crystallizes in the monoclinic space group al Fig.\u00a01.et al., 1990via C\u2014H\u22efO hydrogen bonding and form a tetra\u00admeric ring with two mol\u00adecules each of NIC and CNBA with et al., 1990bc plane O\u2014H\u22efN(NIC) hydrogen bond and additional (NIC)N\u2014H\u22efO(CNBA) and (NIC)C\u2014H\u22efO(CNBA) hydrogen bonds are observed Fig.\u00a02. In thisne Fig.\u00a03.CrystalExplorer and nicotinamide in a minimum amount of ethanol and made up to a volume of 10\u2005ml. Ten different combinations of the mixture were prepared using ethanol\u2013hexane as the solvent mixture over the ratio range 1:1 to 1:10. The mixture was kept in a 5\u2005ml beaker and covered with parafilm, with four to five small holes in it. These solutions were allowed to evaporate slowly at room temperature (27 \u00b0C) over several days to obtain single crystals. After a few days, colourless crystals were obtained from ethanol\u2013hexane solutions with concentration ratios of 1:10, 1:2 and 1:4. The melting point of the obtained crystal was 159.7 \u00b0C.Crystal data, data collection, and structure refinement details are summarized in Table\u00a0210.1107/S2056989019013859/eb2025sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019013859/eb2025Isup2.hklStructure factors: contains datablock(s) I. DOI: 1958621, 1958621CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Four 1,2,4-triazole groups of the ligand link two AgI atoms, as well as AgI and VV centres, forming the heterobimetallic coordination cluster {AgI2(VVO2F2)2(tr)4}. VV exists as a vanadium oxofluoride anion and possesses a distorted trigonal\u2013bipyramidal coordination environment [VO2F2N]. A carb\u00adoxy\u00adlic acid functional group of the ligand stays in a neutral form and is involved in hydrogen bonding with solvent water mol\u00adecules and VO2F2\u2212 ions of adjacent mol\u00adecules. The extended hydrogen-bonding network is responsible for the crystal packing in the structure.The crystal structure of the title mol\u00adecular complex, [Ag It has recently been shown 2(tr)2}2+ binuclear fragment, two N atoms remain uncoordinated and have potential for further inter\u00adactions. In aqueous reaction media, vanadium oxofluorides exist in anionic forms with weakly coordinated water mol\u00adecules that are very labile toward N-donor ligand substitution. Thus, a combination of an AgI\u2013triazole cation and VOF anions lead to the neutral tetra\u00adnuclear {AgI2(VVO2F2)2(tr)4} unit, which was found in the structure of the title [Ag2(VO2F2)2(tr-ad-COOH)4]\u00b74H2O complex I bond lengths , while the other two 1,2,4-triazole groups combine the Ag and V centres [Ag\u22efV= 3.5376\u2005(6)\u2005\u00c5]. The VV atom possesses a distorted trigonal\u2013bipyramidal coordination environment [VO2F2N] with short V\u2014O bonds , V\u2014F bonds and an elongated V\u2014N bond [2.152\u2005(3)\u2005\u00c5]. The polyhedra can be more precisely described by the Reedijk\u2019s factor \u03c4 2(tr)4} is formed. A search in the Cambridge Structural Database 2[Mo4O13]}\u00b72H2O \u2005\u00c5 , while another COOH group, as a hydrogen-bond acceptor, is directed toward the F atom of a {VO2F2} anion . Two water mol\u00adecules are inter\u00adbonded [O2W\u2014H3W\u22efO1W = 2.753\u2005(4)\u2005\u00c5] and additionally act as hydrogen-bond donors with O and F atoms from the neighboring {VO2F2} anions and as hydrogen-bond acceptor (in the case of O2W ) with the O3 atom from an adjacent carb\u00adoxy\u00adlic group. Some weak contacts between the triazole C\u2014H groups and F atoms of the VOF anions are also observed.The structure of k Figs. 2 and 3 \u25b8.tr-ad-COOH) was synthesized in 63% yield by refluxing 3-amino-adamantane-1-carb\u00adoxy\u00adlic acid , tr-ad-COOH , V2O5 and 5\u2005mL of water with aqueous HF was added into a Teflon vessel. Then the components were heated at 423\u2005K for 24\u2005h and slowly cooled to room temperature over 50\u2005h, yielding light-yellow prisms of I .1--3-carb\u00adoxy\u00adadamantane (2), C\u2014H = 0.98\u2005\u00c5 (adamantane CH) and with Uiso(H) = 1.2Ueq(C). O-bound hydrogen atoms were located in a difference-Fourier map and then refined with O\u2014H = 0.82\u2005\u00c5 (carb\u00adoxy\u00adlic) or 0.85\u2005\u00c5 (H2O) with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019006844/zq2246sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019006844/zq2246Isup2.hklStructure factors: contains datablock(s) I. DOI: 1915603CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structures of the title spiro derivatives are described and the analysis of the inter\u00admolecular contacts in the crystals using Hirshfeld surface analysis and two-dimensional fingerprint plots is reported. a-hexa\u00adhydro\u00adspiro\u00adpyrrolo\u00adthia\u00adzole-11,11\u2032-indeno\u00adquinoxaline], C37H26N4O3S, (I), and 6\u2032--6a\u2032-nitro-6\u2032,6a\u2032,6b\u2032,7\u2032,8\u2032,9\u2032,10\u2032,12a\u2032-octa\u00adhydro-2H-spiro\u00adindolizin]-2-one, C36H28N2O4, (II), are new spiro derivatives, in which both the pyrrolidine rings adopt twisted conformations. In (I), the five-membered thia\u00adzole ring adopts an envelope conformation, while the eight-membered pyrrolidine-thia\u00adzole ring adopts a boat conformation. An intra\u00admolecular C\u2014H\u22efN hydrogen bond occurs, involving a C atom of the pyran ring and an N atom of the pyrazine ring. In (II), the six-membered piperidine ring adopts a chair conformation. An intra\u00admolecular C\u2014H\u22efO hydrogen bond occurs, involving a C atom of the pyrrolidine ring and the keto O atom. For both compounds, the crystal structure is stabilized by inter\u00admolecular C\u2014H\u22efO hydrogen bonds. In (I), the C\u2014H\u22efO hydrogen bonds link adjacent mol\u00adecules, forming R22(16) loops propagating along the b-axis direction, while in (II) they form zigzag chains along the b-axis direction. In both compounds, C\u2014H\u22ef\u03c0 inter\u00adactions help to consolidate the structure, but no significant \u03c0\u2013\u03c0 inter\u00adactions with centroid\u2013centroid distances of less than 4\u2005\u00c5 are observed.The title compounds, 6--6a-nitro-6,6a,6\u2005b,7,9,11 The eight-membered pyrrolidine-thia\u00adzole ring (S1/C24\u2013C27/C1/C16/N3) adopts a boat conformation with a total puckering amplitude Q = 1.351\u2005(2)\u2005\u00c5 and \u03c6 = 321.43\u2005(8)\u00b0. The mean planes of the pyran and thia\u00adzole rings are inclined to each other by 77.5\u2005(2)\u00b0. The mean plane of the pyrazine ring (N1/N2/C8/C9/C14/C15) forms a dihedral angle of 57.1\u2005(2)\u00b0 with the mean plane of the pyran ring, while it is almost perpendicular with respect to the mean plane of the pyrrolidine ring, forming an angle of 89.8\u2005(2)\u00b0. The pyrazine ring is inclined by 51.9\u2005(2), 1.9\u2005(2) and 69.5\u2005(2)\u00b0 with respect to the mean planes of the thia\u00adzole and cyclo\u00adpentene ring and the naphthalene ring system, respectively. An intra\u00admolecular C23\u2013H23\u22efN1 hydrogen bond is formed ed Fig.\u00a01.q2 = 0.045\u2005(2)\u2005\u00c5, \u03b8 = 175.7\u2005(2)\u00b0 and \u03c6 = 22\u2005(3)\u00b0. The dihedral angle between the ace\u00adnaphthyl\u00adene (C1\u2013C12) and naphthalene (C27\u2013C36) ring systems is 63.8\u2005(6)\u00b0. Moreover, this moiety is inclined of 85.3\u2005(1), 36.1\u2005(1) and 89.4\u2005(2) \u00b0 with respect to the mean planes of the pyrrolidine (N1/C12/C17\u2013C19), pyran (O4/C18\u2013C20/C25/C26) and piperidine (N1/C13\u2013C17) rings, respectively. The keto atom O1 deviates from the mean plane of the ace\u00adnaphthyl\u00adene unit by 0.148\u2005(1)\u2005\u00c5. An intra\u00admolecular C17\u2014H17\u22efO1 hydrogen bond is present nt Fig.\u00a02.b-axis direction. The loops are linked by C\u2014H\u22efS hydrogen bonds, forming layers parallel to the (101) plane; C\u2014H\u22ef\u03c0 inter\u00adactions are present within the layers on Fig.\u00a04. A C\u2014H\u22ef\u03c0et al., 2007CrystalExplorer17 through white to blue . The red spots on the surface indicate the inter\u00admolecular contacts involved in hydrogen bonding.The Hirshfeld surfaces of (I)b), C\u22efH/H\u22efC , O\u22efH/H\u22efO , S\u22efH/H\u22efS and N\u22efH/H\u22efN , followed by the C\u22efC contacts . For (II)b), C\u22efH/H\u22efC , O\u22efH/H\u22efO , followed by the C\u22efC contacts . In both compounds the H\u22efH inter\u00admolecular contacts predominate.The fingerprint plots for the two compounds are given in Figs. 9et al., 2016H,6\u2032H,6b\u2032H-spiro\u00adindolizin]-2-one skeleton yielded five hits: namely 6-(4- meth\u00adoxy\u00adphen\u00adyl)-6a-nitro-6,6a,6b,7,8,9,10,12a-octa\u00adhydro\u00adspiro-indolizine-12,3-indolin]-2-one to a solution of indeno\u00adquinoxalinone and thia\u00adzolidine-4-carb\u00adoxy\u00adlic acid in dry toluene, 0.302\u2005g (1.0\u2005mmol) of 2--3-nitro-2H-chromene were added under a nitro\u00adgen atmosphere.Compound (II) to a solution of ace\u00adnaphtho\u00adquinone and pipacolinic acid in dry toluene, of 2--3-nitro-2H-chromene were added under a nitro\u00adgen atmosphere.The solutions were refluxed for 18\u2005h in a Dean\u2013Stark apparatus to give the cyclo\u00adadducts. After completion of the reactions as indicated by TLC, the solvent was evaporated under reduced pressure. The crude products obtained were purified by column chromatography using hexa\u00adne/EtOAc (7:3) as eluent (yield 84%). Colourless block-like crystals of the title compounds, suitable for X-ray diffraction analysis, were obtained by slow evaporation of solutions in ethanol.Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq for all other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S205698901901291X/xi2018sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S205698901901291X/xi2018Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S205698901901291X/xi2018IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S205698901901291X/xi2018Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901901291X/xi2018IIsup5.cmlSupporting information file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The dihedral angle between the 4-bromo\u00adphenyl rings is 51.56\u2005(2)\u00b0. In the crystal, mol\u00adecules are linked into a zigzag chain propagating along [001] by weak C\u2014H\u22ef\u03c0 inter\u00adactions. The conformations of related bis\u00adchalcones are surveyed and a Hirshfeld surface analysis is used to investigate and qu\u00adantify the inter\u00admolecular contacts.In the title bis\u00adchalcone, C E,4E)-1,5-di\u00adphenyl\u00adpenta-1,4-dien-3-one] was first prepared by the base-catalyzed Aldol condensation of benzaldehyde and acetone \u2014C=C\u2014) moiety. Bischalcones have a number of uses including anti-inflammatory ; \u03c44 (C13\u2014C12\u2014C11\u2014C10)] and the carbonyl double bond \u00b0; \u03c44 = \u22126.2\u2005(4)\u00b0] but the conformations of the olefinic double bonds are very different: one is in s-trans [\u03c42 = 160.7\u2005(3)\u00b0] conformation and in s-cis [\u03c43 = \u221215.2\u2005(4)\u00b0] conformation with the central C=O double bond. These torsions result in an overall twisted shape for (I)The asymmetric unit of (I))] Fig.\u00a02. The 4-bvia C1\u2014H1A\u22efCg1i . The C14\u2014H14A\u22efCg2ii inter\u00adactions lead to a zigzag chain along the c-axis direction.No classical hydrogen bonding is possible in (I)1i Fig.\u00a03a. The Cii Fig.\u00a03b inter\u00adet al., 2016E,4E)-1,5-di\u00adphenyl\u00adpenta-1,4-dien-3-one as the main skeleton revealed the presence of 27 structures containing a similar bis\u00adchalcone moiety to the title compound but with different substituents on the terminal phenyl rings. The different substituents (1R and 2R) together with the torsion angles of the penta-4,4-dien-3-one connecting bridge are compiled in Table\u00a02s-cis or s-trans), the olefinic double bonds are close to coplanar with their attached phenyl rings as indicated by their \u03c41 and \u03c44 torsion angles, which fall in the range of 0.0\u201317.8\u00b0, except for the compounds AMEXUN and HUDLEY, which have somewhat larger \u03c41 and \u03c44 values of 22.5\u201327.4\u00b0. The olefinic double bonds for the symmetrical compounds are mostly in s-cis conformations with the carbonyl double bond (\u03c42/\u03c43 torsion angles of 0.1\u201321.9\u00b0). However, both the olefinic double bonds of compounds GOLGOD and GOLGOD02 are in s-trans conformations with the carbonyl double bond (\u03c42/\u03c43 = 152.2\u2013153.4\u00b0). Furthermore, it may be noted that the symmetrical conformation at the penta-4,4-dien-3-one connection bridge is not affected by the different substituents at the 1R and 2R positions in EDUSEE, SAFZOO and XOHVUN. Most of the unsymmetrical compounds (one C=C\u2014C=O bond s-cis and one s-trans) have \u03c41 and \u03c44 values of 0.5\u201317.2\u00b0, which indicates that the olefinic double bonds are close to coplanar to their attached phenyl ring. The outliers are MESXEQ and WIHBUL, which have \u03c41 and \u03c44 values of 18.2\u201351.8\u00b0 and 21.4\u201351.8\u00b0, respectively. The torsion angles \u03c42 and \u03c43 for the unsymmetrical compounds, including (I)s-trans and s-cis conformations between the olefinic double bonds and the carbonyl double bond.A survey of the Cambridge Structural Database CrystalExplorer17.5 correspond to the C14\u2014H14A\u22efCg2ii inter\u00adaction. Even through the C1\u2014H1A\u22efCg1i inter\u00adaction is not visible in the dnorm surface mapping, this inter\u00adaction can be seen as a unique pattern of a red \u2018circle\u2019 on the shape-index surface mapping . Besides the C\u2014H\u22ef\u03c0 inter\u00adactions, the dnorm surface mapping indicated a short contact between atom O1 and C5 with a distance of 0.06\u2005\u00c5 shorter than the sum of the van der Waals radii of O and C atoms . Together with this short contact, another weak C7\u2014H7A\u22efO1 inter\u00adaction was also revealed as light spots on the dnorm surface .The Hirshfeld surfaces mapped with normalized contact distance rm Fig.\u00a04a corresng Fig.\u00a04b. Besidce Fig.\u00a05b.de and di diagonal axes. The H\u22efC/C\u22efH contacts are the most populated contacts and contribute 34.1% to the total inter\u00admolecular contacts, followed by H\u22efH (22.1%), H\u22efBr/Br\u22efH (20.4%) and H\u22efO/O\u22efH (9.2%) contacts . The H\u22efH contacts appear in the central region of the fingerprint plots with de = di = 2.4\u2005\u00c5 . With the presence of relatively larger bromine atoms in the structure, the H\u22efBr/Br\u22efH contacts appear as symmetrical broad wing at diagonal axes of de + di \u2243 3.0\u2005\u00c5 . Two symmetric spikes in the fingerprint plots with a short spike at de + di \u2243 2.7\u2005\u00c5 represent the H\u22efO/O\u22efH contacts , indicating the presence of the weak C7\u2014H7A\u22efO1 inter\u00adaction. The percentage contributions for other contacts are less than 15% in the Hirshfeld surface mapping.As illustrated in Fig.\u00a06ts Fig.\u00a06. As the \u2005\u00c5 Fig.\u00a06b. The H\u2005\u00c5 Fig.\u00a06c. With \u2005\u00c5 Fig.\u00a06d. Two sts Fig.\u00a06e, indicA mixture of 4-bromo\u00adbenzaldehyde and acetone dissolved in absolute ethanol (30\u2005ml) was slowly added to an aqueous solution of potassium hydroxide (4.0\u2005g in 20\u2005ml water). The mixture was vigorously stirred at room temperature for two\u2005h and then 20\u2005ml chilled water was added. The resulting yellow precipitate was recovered by vacuum filtration and washed with cold water (100\u2005ml). The crude product was recrystallized from absolute ethanol solution as yellow blocks.-1,5-Bis\u00ad(4-bromo\u00adphen\u00adyl)penta-1,4-dien-3-one; pure yellow solid , m.p. 484\u2005K; IR \u03bdmax 594, 687, 813, 979, 1066, 1181, 1320, 1398, 1480, 1581, 1643\u2005cm\u22121, UV\u2013Vis \u03bbmax. 227 and 317\u2005nm, 1H NMR:\u03b4H 7.02 , 7.45 , 7.54 , 7.67 ; 13C NMR:\u03b4C 124.62, 125.69, 129.25, 131.74, 133.29, 141.83, 188.22; HRMS (ES): MH+, found: 392 C17H12Br2O+ requires: 391.92.Uiso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019006480/hb7821sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019006480/hb7821Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019006480/hb7821Isup3.cmlSupporting information file. DOI: 1914420CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the compound, the acid and base mol\u00adecules are linked by a short hydrogen bond [O\u22efO = 2.4393\u2005(15)\u2005\u00c5], in which the H atom is disordered over two positions with equal occupancies.The title compound was analysed as a disordered structure over two states, 9H7.5NO\u00b7C7H3.5ClNO4, was analysed as a disordered structure over two states, viz. co-crystal and salt, accompanied by a keto\u2013enol tautomerization in the base mol\u00adecule. The co-crystal is 4-chloro-2-nitro\u00adbenzoic acid\u2013quinolin-4(1H)-one (1/1), C7H4ClNO4\u00b7C9H7NO, and the salt is 4-hy\u00addroxy\u00adquinolinium 4-chloro-2-nitro\u00adbenzoate, C9H8NO+\u00b7C7H3ClNO4\u2212. In the compound, the acid and base mol\u00adecules are held together by a short hydrogen bond [O\u22efO = 2.4393\u2005(15)\u2005\u00c5], in which the H atom is disordered over two positions with equal occupancies. In the crystal, the hydrogen-bonded acid\u2013base units are linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, forming a tape structure along the a-axis direction. The tapes are stacked into a layer parallel to the ab plane via \u03c0\u2013\u03c0 inter\u00adactions [centroid\u2013centroid distances = 3.5504\u2005(8)\u20133.9010\u2005(11)\u2005\u00c5]. The layers are further linked by another C\u2014H\u22efO hydrogen bond, forming a three-dimensional network. Hirshfeld surfaces for the title compound mapped over shape-index and dnorm were generated to visualize the inter\u00admolecular inter\u00adactions.The title compound, C D\u2014H\u22efA hydrogen bonding in chloro- and nitro-substituted benzoic acid\u2013pyridine derivative systems, we have shown that several compounds, namely, three compounds of quinoline with 3-chloro-2-nitro\u00adbenzoic acid, 4-chloro-2-nitro\u00adbenzoic acid and 5-chloro-2-nitro\u00adbenzoic acid -quinolinone (1/1), and the salt, 4-hy\u00addroxy\u00adquinolinium 4-chloro-2-nitro\u00adbenzoate, accompanied by a keto\u2013enol tautomerization in the base mol\u00adecule. The C10\u2014O5 bond length [1.2956\u2005(18)\u2005\u00c5] is inter\u00admediate between a C\u2014O single bond [1.36\u2005\u00c5 in phenol] and a C=O double bond [1.23\u2005\u00c5 in ketones of the (Car)2\u2014C=O type] of the acid mol\u00adecule and the quinoline ring system (N2/C8\u2013C16) of the base are slightly inclined to each other by a dihedral angle of 10.27\u2005(6)\u00b0, while the carb\u00adoxy group (O1/C7/O2) is twisted by 38.66\u2005(18) and 45.93\u2005(18)\u00b0, respectively, with respect to the C1\u2013C6 ring and the N2/C8\u2013C16 ring system. The dihedral angle between the C1\u2013C6 ring and the nitro group (O3/N1/O4) is 50.33\u2005(19)\u00b0.i, C8\u2014H8\u22efO5i and C9\u2014H9\u22efO1i; symmetry code as in Table\u00a01a axis \u2212x\u00a0+\u00a0y\u00a0\u2212\u00a0z]. The layers are further linked by another C\u2014H\u22efO hydrogen bond \u2013(3)]. The \u03c0\u2013\u03c0 inter\u00adactions between the benzene rings of the acid mol\u00adecules [Cg1\u22efCg1iv] and between the quinoline ring systems of the base mol\u00adecules are indicated by blue and red triangles on the shape-index surfaces [arrows (4) and (5)].In order to visualize the inter\u00admolecular inter\u00adactions, Hirshfeld surfaces for the acid and base mol\u00adecules of the title compound, mapped over shape-index and i Table\u00a01 are repret al., 2016H)-quinolinone (keto tautomer) showed one structure, namely, 4-amino-1-(2-(hy\u00addroxy\u00admeth\u00adyl)-1,3-oxa\u00adthio\u00adlan-5-yl)-2(1H)-pyrimidinone 4(1H)-quinolinone -quinolinone itself was reported by Nasiri et al. of 4-hy\u00addroxy\u00adquinoline (0.075\u2005g) with 4-chloro-2-nitro\u00adbenzoic acid (0.106\u2005g) in a 1:1 molar ratio at room temperature.A (O1 site) and H1B (O5 site), respectively, with bond restraints of O\u2014H = 0.84\u2005(1)\u2005\u00c5 and with Uiso(H) = 1.5Ueq(O). In the final refinement, the occupancies were fixed at 0.5, and one outlier was omitted. The N-bound H atom was refined freely [refined distance: N2\u2014H2 = 0.89\u2005(2)\u2005\u00c5]. Other H atoms were positioned geometrically (C\u2014H = 0.95\u2005\u00c5) and treated as riding, with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901901497X/lh5935sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S205698901901497X/lh5935Isup2.hklStructure factors: contains datablock(s) I. DOI: 1963942, 1963942CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The chains are linked into layers parallel to the bc plane by sets of four C\u2014H\u22efO hydrogen bonds and the layers are tied together by complementary \u03c0-stacking inter\u00adactions.The di\u00adhydro\u00adqinoxalinone portion of the mol\u00adecule is planar to within 0.0512\u2005(12)\u2005\u00c5. In the crystal, a combination of C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds together with slipped \u03c0-stacking and C\u2014H\u22ef\u03c0(ring) inter\u00adactions lead to the formation of chains extending along the 16H17N5O3, is build up from two fused six-membered rings linked to a 1,2,3-triazole ring, which is attached to an ethyl azido-acetate group. The di\u00adhydro\u00adqinoxalinone portion is planar to within 0.0512\u2005(12)\u2005\u00c5 and is oriented at a dihedral angle of 87.83\u2005(5)\u00b0 with respect to the pendant triazole ring. In the crystal, a combination of inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efN hydrogen bonds together with slipped \u03c0-stacking [centroid\u2013centroid distance = 3.7772\u2005(12)\u2005\u00c5] and C\u2014H\u22ef\u03c0 (ring) inter\u00adactions lead to the formation of chains extending along the c-axis direction. Additional C\u2014H\u22efO hydrogen bonds link these chains into layers parallel to the bc plane and the layers are tied together by complementary \u03c0-stacking [centroid\u2013centroid distance = 3.5444\u2005(12)\u2005\u00c5] inter\u00adactions. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (44.5%), H\u22efO/O\u22efH (18.8%), H\u22efN/N\u22efH (17.0%) and H\u22efC/C\u22efH (10.4%) inter\u00adactions.The mol\u00adecule of the title compound, C The pendant triazole ring is inclined to this plane by 87.83\u2005(5)\u00b0.Dhyqnx\u22efOEthazac, C\u2014HEthazac\u22efODhyqnx, C5\u2014HDhyqnx\u22efNEthazac and C\u2014HTrz\u22efNDhyqnx hydrogen bonds rings [centroid\u2013centroid distance = 3.7772\u2005(12)\u2005\u00c5] and by complementary C\u2014HDhyqnx\u22efCg3 inter\u00adactions [Cg3 is the centroid of the benzene ring B (C1\u2013C6)] (Table\u00a01bc plane by sets of four C\u2014HDhyqnx\u22efOEthazac hydrogen bonds (Table\u00a01a-axis direction by inversion-related slipped \u03c0-stacking inter\u00adactions between the A and B rings [centroid\u2013centroid distance = 3.5444\u2005(12)\u2005\u00c5] analysis contacts -one (0.65\u2005mmol) in ethanol (20\u2005mL) was added ethyl azido\u00adacetate (1.04\u2005mmol). The mixture was stirred under reflux for 24\u2005h. After completion of the reaction (monitored by TLC), the solution was concentrated and the residue was purified by column chromatography on silica gel by using as eluent a hexa\u00adne/ethyl acetate (9/1) mixture. Crystals were obtained when the solvent was allowed to evaporate. The solid product isolated was recrystallized from ethanol to afford yellow crystals in 75% yield.To a solution of 3-methyl-1-(prop-2-yn\u00adyl)-3,4-di\u00adhydro\u00adquinox\u00adalin-2 global, I. DOI: 10.1107/S2056989018014561/xu5945Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018014561/xu5945Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018014561/xu5945Isup4.cmlSupporting information file. DOI: 1873385CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In each of the title compounds, the packing is driven by C\u2014H\u22efF inter\u00adtactions, along with a variety of C\u2014H\u22efO, C\u2014O\u22ef\u03c0, and C\u2014F\u22ef\u03c0 contacts. Hirshfeld surface analyses were conducted to aid in the visualization of these various influences on the packing. 14H8F4N2O2 and C14H8F4N2O3, are reported. In each crystal, the packing is driven by C\u2014H\u22efF inter\u00adtactions, along with a variety of C\u2014H\u22efO, C\u2014O\u22ef\u03c0, and C\u2014F\u22ef\u03c0 contacts. Hirshfeld surface analysis was conducted to aid in the visualization of these various influences on the packing: they showed that the largest contributions to the surface contacts arise from H\u22efF/F\u22efH inter\u00adactions, followed by H\u22efH and O\u22efH/H\u22efO.The syntheses and crystal structures of the title compounds, C In mol\u00adecule A, the N1/C1\u2013C5 plane is rotated by 58.05\u2005(5)\u00b0 relative to the N2/C6/C7/C12/C13 plane and the corres\u00adponding dihedral angle for mol\u00adecule B is 61.65\u2005(7)\u00b0. The addition of an oxygen atom between N2 and C3 in II alleviates this steric restriction and only one 19F NMR peak in solution is observed for the ortho-F atoms; even so, the dihedral angle between the N1/C1\u2013C5 and N2/C6/C7/C12/C13 planes in the crystal of II of 84.01\u2005(5)\u00b0 is larger than those found in I.Compound it Fig.\u00a01. The synI and II are of the type C\u2014H\u22efO, C\u2014H\u22efF, C\u2014O\u22ef\u03c0, and C\u2014F\u22ef\u03c0 and the olefinic hydrogen atoms . A weak inter\u00adaction is also observed for a bridge hydrogen atom in II, C14\u2014H14B\u22efF4iii. The packing is further aided by \u03c0-inter\u00adactions with the pyridine ring .The main directional inter\u00adactions in the crystal structures of Tables 1 and 2 \u25b8.CrystalExplorer17.5 to 1.3800 (blue) a.u. The pale-red spots symbolize short contacts and negative dnorm values on the corresponding surface plots shown in Fig.\u00a02Hirshfeld surface analysis < 1.25\u2005\u00c5 in I and 1.10\u2005\u00c5 < (di + de) < 1.35\u2005\u00c5 in II. H\u22efH contacts make the second largest contribution (20.2% in I and 14.1% in II), shown in the middle region 1.10\u2005\u00c5 < (di + de) < 1.18\u2005\u00c5 in I and II. The third largest contribution is from O\u22efH/H\u22efO contacts. In I, the corresponding spike is partially overlapped with the spike representing F\u22efH/H\u22efF contacts, appearing at 1.05\u2005\u00c5 < (di + de) < 1.40\u2005\u00c5. The O\u22efH/H\u22efO spike is clearly visible in the fingerprint plot of II, shown in the region of 1.10\u2005\u00c5 < (di + de) < 1.40\u2005\u00c5.The largest contribution to the overall crystal packing in both compounds is from F\u22efH/H\u22efF inter\u00adactions -dione-based compounds with an aromatic substituent on the nitro\u00adgen atom yielded 58 results. The dihedral angle between the aromatic ring plane and the succinimide plane is bimodally distributed between 43 and 90\u00b0, with peaks near 60 and 75\u00b0.A search of the November 2018 release of the Cambridge Structure Database (Groom Synthesis of (I) penta\u00adfluoro\u00adpyridine , -3a,4,7,7a-tetra\u00adhydro-1H-4,7-methano\u00adiso\u00adindole-1,3(2H)-dione , and tri\u00adethyl\u00adamine were combined in DMF (150\u2005ml). The resulting solution was stirred at room temperature for 24\u2005h. Diethyl ether (150\u2005ml) and saturated aqueous ammonium chloride (100\u2005ml) were added and the biphasic solution stirred vigorously for 2\u2005h. The organic layer was separated and the remaining aqueous portion extracted with diethyl ether (2 \u00d7 150\u2005ml). The combined organic fractions were washed with water (2 \u00d7 1\u2005l) and brine (2 \u00d7 300\u2005ml), dried over MgSO4, and the solvent removed via rotary evaporation. The resulting off-white solid was dissolved in refluxing EtOH (20\u2005ml) and cooled to 278\u2005K for 12\u2005h. Vacuum filtration, washing with cold EtOH (20\u2005ml), and vacuum drying afforded the target compound as a white, crystalline solid . 1H NMR : 6.28 , 3.58 , 3.54 , 1.74 .19F NMR : \u221290.5 (2F), \u2212141.7 (1F), \u2212143.1 (1F).Synthesis of (II) to a stirred solution of potassium carbonate , -2-hy\u00addroxy-3a,4,7,7a-tetra\u00adhydro-1H-4,7-methano\u00adiso\u00adindole-1,3(2H)-dione , penta\u00adfluoro\u00adpyridine , and 140\u2005ml of DMF were added. The resulting solution was stirred at room temperature for 24\u2005h. Diethyl ether (150\u2005ml) and saturated aqueous ammonium chloride (100\u2005ml) were added and the biphasic solution stirred vigorously for 2\u2005h. The organic layer was separated and the remaining aqueous portion extracted with diethyl ether (2 \u00d7 150\u2005ml). The combined organic fractions were washed with water (2 \u00d7 1\u2005l) and brine (2 \u00d7 300\u2005ml), dried over MgSO4, and the solvent removed via rotary evaporation. The resulting off-white solid was dissolved in refluxing EtOH (50\u2005ml) and cooled to 278\u2005K for 12\u2005h. Vacuum filtration, washing with cold EtOH (20\u2005ml) and vacuum drying afforded the target compound as a white, crystalline solid . 1H NMR : 6.23 , 3.50 , 3.34 , 1.68 .19F NMR : \u221287.4 (2F), \u2212156.3 (2F).Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989019009769/hb7832sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989019009769/hb7832Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019009769/hb7832IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989019009769/hb7832Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019009769/hb7832IIsup5.cmlSupporting information file. DOI: 1879244, 1885019CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound crystallizes with two independent mol\u00adecules in the asymmetric unit. In the crystal, mol\u00adecules are linked by N\u2014H\u22efO hydrogen bonds, C\u2014H\u22ef\u03c0, I\u22efS and I\u22efI inter\u00adactions into a three-dimensional network. A and B) are present in the asymmetric unit of the title compound, C11H9IN2OS, which differ mainly in the dihedral angle between the phenyl and thia\u00adzole rings . In the crystal, the mol\u00adecules form \u22efA\u22efB\u22efA\u22efB\u22ef chains along the [001] and [010] directions through moderate N\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adactions, respectively. The overall three-dimensional network is formed by I\u22efI and I\u22efS inter\u00adactions. Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from H\u22efC/C\u22efH (26.2%), H\u22efH (20.9%), H\u22efI/I\u22efH (19.4%) and H\u22efO/O\u22efH (6.8%) inter\u00adactions.Two crystallographically independent mol\u00adecules ( Unlike the related compound 2-acetamido-4-p-tolyl-1,3-thia\u00adzole .The title 2-aceto\u00adamido\u00adthia\u00adzole derivative crystallizes in the monoclinic space group it Fig.\u00a01. The privia a C(4) synthon contacts [3.7758\u2005(9)\u2005\u00c5] contacts of type I [\u03b81 = \u03b82 = 146.91\u2005(8)\u00b0] with a length of 3.8547\u2005(5)\u2005\u00c5.In the crystal, mol\u00adecules are linked by N1\u2014H1\u22efO2 and N3\u2014H3\u22efO1 moderate hydrogen bonds n Table\u00a01, forming\u00c5] Fig.\u00a03. AdditioCrystal Explorer17.5 to 1.3937 (blue) a.u. . The pair of characteristic wings in this fingerprint plot corresponds to the C\u2014H\u22ef\u03c0 inter\u00adactions between the phenyl groups . The H\u22efH and H\u22efI/I\u22efH contacts make similar contributions to the total Hirshfeld surface of 20.9 and 19.4%, respectively. The reciprocal H\u22efO/O\u22efH inter\u00adactions (6.8%) are seen as sharp symmetrical spikes with tips at ed + id \u223c1.9\u2005\u00c5 and arising from the N\u2014H\u22efO hydrogen bond . Inter\u00admolecular I\u22efS/S\u22efI and I\u22efI inter\u00adactions make smaller contributions to the Hirshfeld surface .A Hirshfeld surface analysis was carried out using u. Fig.\u00a04. The intps Fig.\u00a05b. The Hts Fig.\u00a05c and 5dnd Fig.\u00a05e. Inter\u22efI Fig.\u00a05f and I\u22efet al., 2016N--4-chloro\u00adbutanamide -1,3-thia\u00adzol-2-yl)-4-chloro\u00adbutanamide. In this compound the dihedral angle is smaller [8.8\u2005(3)\u00b0] as a result of an intra\u00admolecular C\u2014H\u22efBr hydrogen bond. In the crystals of these compounds, only 5,5\u2032-di\u00adbromo-4,4\u2032-bis(penta\u00adfluoro\u00adphen\u00adyl)-2,2\u2032-bi-1,3-thia\u00adzole exhibits a type II halogen\u2013halogen inter\u00adaction with a Br\u22efBr distance of 3.6777\u2005(3)\u2005\u00c5 and angles of 68.88\u2005(5) and 174.77\u2005(5)\u00b0.A search of the Cambridge Structural Database acetamide and iodine was placed in an open vessel containing a Teflon-coated stir bar. The mixture was dissolved in 3\u2005mL of ethanol and the vessel was placed in the microwave cavity and subjected to MW irradiation (150\u2005W) for 60\u2005min, at 363\u2005K and a pressure of 2\u2005psi. The reaction mixture was then cooled at room temperature and 5\u2005mL of NH4OH were added. The obtained mixture was dissolved in ethyl acetate (50\u2005mL) and washed with brine (3\u00d7). The organic layer was separated, dehydrated with Na2SO4, and evaporated in vacuo until dryness. The product was purified by flash column chromatography with a mixture of petrol\u2013di\u00adchloro\u00admethane\u2013acetone (5:3:2). The title compound was obtained as pale-yellow needles in 30% yield . A diluted solution of the compound was prepared in hexane and kept on a dry and dark place at room temperature. Crystals were obtained after one week of slow evaporation. Spectroscopic data: 1H NMR : 11.37 , 7.80 , 7.43 , 1.62 . 13C NMR : 168.8 (s), 163.6 (s), 151.4 (s), 134.5 (s), 129.0 (d), 128.9 (d), 128.7 (d), 62.4 (s) 21.9 (c).A mixture of Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019004791/lh5897sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019004791/lh5897Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019004791/lh5897Isup3.cmlSupporting information file. DOI: 1908908CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I (HypSiBr), and II (TipSiBr), crystallized in the cubic space group PaPThe title compounds, 9H27BrSi4 and C27H63BrSi4 , are described. Compound I was prepared in 85% yield by free-radical bromination of 1,1,1,3,3,3-hexa\u00admethyl-2-(tri\u00admethyl\u00adsil\u00adyl)tris\u00adilane using bromo\u00adbutane and 2,2\u2032-azobis(2-methyl\u00adpropio\u00adnitrile) as a radical initiator at 333\u2005K. The mol\u00adecule possesses threefold rotational symmetry, with the central Si atom and the Br atom being located on the threefold rotation axis. The Si\u2014Br bond distance is 2.2990\u2005(12)\u2005\u00c5 and the Si\u2014Si bond lengths are 2.3477\u2005(8)\u2005\u00c5. The Br\u2014Si\u2014Si bond angles are 104.83\u2005(3)\u00b0 and the Si\u2014Si\u2014Si bond angles are 113.69\u2005(2)\u00b0, reflecting the steric hindrance inherent in the three tri\u00admethyl\u00adsilyl groups attached to the central Si atom. Compound II was prepared in 55% yield by free-radical bromination of 1,1,1,3,3,3-hexa\u00adisopropyl-2-(triiso\u00adpropyl\u00adsil\u00adyl)tris\u00adilane using N-bromo\u00adsuccinimide and 2,2\u2032-azobis(2-methyl\u00adpropio\u00adnitrile) as a radical initiator at 353\u2005K. Here the Si\u2014Br bond length is 2.3185\u2005(7)\u2005\u00c5 and the Si\u2014Si bond lengths range from 2.443\u2005(1) to 2.4628\u2005(9)\u2005\u00c5. The Br\u2014Si\u2014Si bond angles range from 98.44\u2005(3) to 103.77\u2005(3)\u00b0, indicating steric hindrance between the three triiso\u00adpropyl\u00adsilyl groups.The synthesis and crystal structures of two tris\u00adsilyl bromide compounds, C The Si1\u2014Br1 bond length is 2.2990\u2005(12)\u2005\u00c5. As for Si2, the C\u2014Si2\u2014C bond angles range from 107.1\u2005(2) to 110.55\u2005(17)\u00b0, while the C\u2014Si2\u2014Si1 bond angles range from 108.61\u2005(10) to 110.16\u2005(11)\u00b0.The mol\u00adecular structure of compound II (TipSiBr), is shown in Fig.\u00a02P4 descriptor for fourfold coordination of 0.90. The Br1\u2014Si1\u2014Si2/Si3/Si4 bond angles range from 98.44\u2005(3) to 103.77\u2005(3)\u00b0, and the Si1\u2014Br1 bond distance is 2.3185\u2005(7)\u2005\u00c5, which is longer than that of compound I [2.2990\u2005(12)\u2005\u00c5]. The \u03c44 descriptor values for atoms Si2, Si3 and Si4 (the silicon atoms of the triiso\u00adpropyl\u00adsilyl groups) are 0.96, 0.97 and 0.95, respectively, indicating that their coordination geometry is closest to an ideal tetra\u00adhedron.The asymmetric unit of compound I or II. Compound II, however, contains four intra\u00admolecular C\u2014H\u22efBr hydrogen bonds (Table\u00a02D\u22efA distances that range from from 3.584\u2005(3) to 3.726\u2005(3)\u2005\u00c5, and D\u2014H\u22efA bond angles that range from 131 to 155\u00b0.There are no significant inter\u00admolecular contacts, other than weak van der Waals inter\u00adactions, present in the crystals of compounds s Table\u00a02. These het al., 20163Si group. Of these, there are only 42 structures where the central silicon atom is bonded directly to a halogen.The Cambridge Structural Database . The protio derivative (HypSiH) is a liquid at room temperature, and the structure of the iodo derivative (HypSiI) has not been deposited in the CSD.For compounds IV (TipSiH) was not found in the CSD, but the journal article 3SiBr, viz. compound II (TipSiBr). Like compounds I and III, compound IV crystallizes in the cubic space group PaiPr3)Si\u2013 groups to push further away from one another, resulting in Si2\u2014Si1\u2014Si2i,ii bond angles of 117.9\u2005(1)\u00b0 and Si2i,ii\u2014Si1\u2014H bond angles of 98.3\u2005(1)\u00b0 . In II, the corresponding Si\u2014Si\u2014Si bond angles range from 115.02\u2005(4) to 116.59\u2005(4)\u00b0 and the Si\u2014Si\u2014Br bond angles vary from 98.44\u2005(3) to 103.77\u2005(3)\u00b0.The X-ray data for compound Compound I: Tris(tri\u00admethyl\u00adsil\u00adyl)silane was added to an oven-dried nitro\u00adgen-flushed 250\u2005ml Schlenk flask with a stir-bar. Bromo\u00adbutane was filtered through a plug of silica gel in a Pasteur pipette and was transferred into the Schlenk flask. AIBN was then added to the flask, and the reaction was heated to 333\u2005K using an oil bath and then heating was stopped. After stirring the reaction overnight at room temperature, GC\u2013MS analysis of a sample indicated incomplete reaction and more AIBN (11\u2005mg) was added to the flask. The reaction was heated once more to 333\u2005K for 1\u2005h. Analysis by GC\u2013MS now indicated that the reaction was complete. The flask was placed in a freezer at 243\u2005K and colourless block-like crystals of I formed overnight. Removal of the solvent in vacuo yielded 2.2\u2005g (85%). 1H NMR \u03b4 0.24 ; 13C NMR \u03b4 \u22120.51 ppm; GC\u2013MS: 11.24\u2005min, m/z = 328, base peak: 73.Compound II: Tris(triiso\u00adpropyl\u00adsil\u00adyl)silane was dissolved in freshly distilled benzene (10\u2005ml) along with NBS (45\u2005mg) and AIBN . The mixture was heated using an oil bath at 333\u2005K for 30\u2005min, when GC\u2013MS analysis indicated that no reaction had occurred. At this point the solution was heated with a heat gun until the reaction mixture turned slightly yellow. The yellow colour dissipated in less than 1\u2005min. Analysis of the reaction mixture by 1H NMR indicated that only 60% of the starting material had been consumed. An additional amount of NBS was added to the reaction flask, and the solution was again heated with a heat gun. The product was isolated by removing the solvent in vacuo and extracting the product from the crude reaction mixture with pentane. The pentane solution was filtered through glass wool, concentrated and weighed (135\u2005mg). Analysis of the product with 1H NMR indicated this was 90% pure. The product was further purified by dissolving this solid in 1\u2005ml pentane, cooling to 195\u2005K and isolating the colourless needle-like crystals of II by removing the solvent with a syringe, washing with pentane and drying in vacuo . 1H NMR \u03b4 1.34 , 1.66 ; 13C NMR \u03b4 16.4, 21.6; HRMS for C17H63BrSi4 calculated 535.2642 (M\u00a0\u2212\u00a0C3H7), found 535.2641.Uiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018009696/su5451sup1.cifCrystal structure: contains datablock(s) I, II, Global. DOI: 10.1107/S2056989018009696/su5451Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989018009696/su5451IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989018009696/su5451Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018009696/su5451IIsup5.cmlSupporting information file. DOI: 1854536, 1854535CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound comprises four crystallographically different mol\u00adecules that are composed of a 1,5-bis\u00ad(4-meth\u00adoxy\u00adphen\u00adyl)penta-1,4-dien-3-ylidenyl group and a benzyl ring connected by a hydrazine-1-carbodi\u00adthio\u00adate bridge. In the crystal, mol\u00adecules are connected into a three-dimensional network through C\u2014H\u22efO, N\u2014H\u22efS and C\u2014H\u22ef\u03c0 inter\u00adactions. 27H26N2O2S2, the asymmetric unit is comprised of four mol\u00adecules (Z = 8 and Z\u2032 = 4). The 4-meth\u00adoxy\u00adphenyl rings are slightly twisted away from their attached olefinic double bonds [torsion angles = 5.9\u2005(4)\u201319.6\u2005(4)\u00b0]. The azomethine double bond has an s-trans configuration relative to one of the C=C bonds and an s-cis configuration relative to the other . The torsion angles between the azomethine C=N double bond and hydrazine-1-carbodi\u00adthio\u00adate moiety indicate only small deviations from planarity, with torsion angles ranging from 0.9\u2005(3) to 6.9\u2005(3)\u00b0 and from 174.9\u2005(3) to 179.7\u2005(2)\u00b0, respectively. The benzyl ring and the methyl\u00adenesulfanyl moiety are almost perpendicular to each other, as indicated by their torsion angles [range 93.7\u2005(3)\u2013114.6\u2005(2)\u00b0]. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO, N\u2014H\u22efS and C\u2014H\u22ef\u03c0(ring) hydrogen-bonding inter\u00adactions into a three-dimensional network. Structural details of related benzyl hydrazine-1-carbodi\u00adthio\u00adate are surveyed and compared with those of the title compound.In the title hydrazinecarbodi\u00adthio\u00adate derivative, C S-benzyl and S-alkyl di\u00adthio\u00adcarbaza\u00adtes are inter\u00adesting ligands in coordination chemistry because they can act as N,S-chelating agents because of the presence of soft sulfur and hard nitro\u00adgen donor atoms than their transition-metal complexes . Structural details of (I) are compared with other hydrazinecarbodi\u00adthio\u00adates.Encouraged by previous findings on various properties of related Schiff base derivatives, we report herein the synthesis and structure determination of the title compound (I) consists of four mol\u00adecules, denoted as A, B, C and D. Orientational disorder of the 1,5-bis(phen\u00adyl)penta-1,4-dien-3-ylidenyl amine moiety in mol\u00adecule B and of the phenyl\u00admethyl moiety in mol\u00adecule D were observed. The mol\u00adecules are composed of a 1,5-bis\u00ad(4-meth\u00adoxy\u00adphen\u00adyl)penta-1,4-dien-3-ylidenyl moiety, connected to a benzyl ring by a hydrazine-1-carbodi\u00adthio\u00adate (\u2013C=N\u2014(NH)\u2014(C=S)\u2014S\u2014C\u2013) bridge ; \u03c46 (C16\u2014C15\u2014O2\u2014C27)], between the 4-meth\u00adoxy\u00adphenyl ring and the olefinic double bond , and between the olefinic double bond and the azomethine double bond . Torsion angles \u03c41 and \u03c46 are approximately 0\u00b0 or \u00b1180\u00b0 in the majority of the four mol\u00adecules, except mol\u00adecule C which has a \u03c41 of 20.9\u2005(3)\u00b0 (Table\u00a01\u03c42 = 5.9\u2005(4)\u201316.4\u2005(13)\u00b0; \u03c45 = 7.5\u2005(3)\u201319.6\u2005(4)\u00b0]. The orientations of the the azomethine double bond with its neighbouring olefinic double bonds relative to the inter\u00admediate C\u2014C bond are different: one is in s-trans [\u03c43 = 147.4\u2005(6)\u2013175.7\u2005(2)\u00b0] conformation and the other in s-cis [\u03c44 = 15.3\u2005(3)\u201337.4\u2005(7)\u00b0] conformation. The dihedral angles between two 4-meth\u00adoxy\u00adphenyl rings in an individual mol\u00adecule are in the range 23.59\u2005(12)\u201389.6\u2005(5)\u00b0 (Table\u00a02\u03c47 (C9\u2014N1\u2014N2\u2014C18), \u03c48 (N1\u2014N2\u2014C18\u2014S1), \u03c49 (N2\u2014C18\u2014S1\u2014C19), \u03c410 (C18\u2014S1\u2014C19\u2014C20) and \u03c411 (S1\u2014C19\u2014C20\u2014C21). In all mol\u00adecules of (I), the hydrazine-1-carbo\u00adthio\u00adate bridges are more or less planar \u20136.9\u2005(3)\u00b0 and 174.9\u2005(3)\u2013179.7\u2005(2)\u00b0, respectively). The torsion angles between the sulfane moiety and the methyl\u00adene moiety indicate a slight twist [\u03c410 = 160.53\u2005(17)\u2013163.36\u2005(16)\u00b0]. These contortions are more severe between the benzyl ring and the methyl\u00adene sulfane moiety where \u03c411 is considerably smaller [\u03c411 = 93.7\u2005(3)\u2013114.6\u2005(2)\u00b0]. The dihedral angles between the benzyl ring and the two 4-meth\u00adoxy\u00adphenyl rings are in the range 31.6\u2005(5)\u201389.9\u2005(8)\u00b0 . In addition, mol\u00adecule C and mol\u00adecule D are connected through C17C\u2014H17C\u22efO1D hydrogen bonds . The four mol\u00adecules are linked into an endless chain parallel to [021] through the combination of these hydrogen bonds hydrazine-1-carbodi\u00adthio\u00adate as reference moiety resulted in 45 structures with different substituents. The reference moiety and relevant torsion angles are illus\u00adtrated in Fig.\u00a061R) together with the torsion angles for the benzyl hydrazine-1-carbo\u00adthio\u00adate moiety in these structures are collated in Table\u00a04\u03c47, \u03c48 and \u03c49; torsion angles \u03c47 and \u03c49 range from 165.1 to 180.0\u00b0 and indicate an anti-periplanar conformation whereas torsion angle \u03c48 is indicative of a syn-periplanar conformation (0.0\u2013 9.1\u00b0). With respect to torsion angle \u03c410, most of the structures adopt an anti-periplanar conformation ranging from 159.5 to 180.0\u00b0, but there are nine structures that adopt either a syn-clinal or an anti-clinal conformation (77.6\u2013110.4\u00b0). In most of the structures, the benzyl ring and the methyl\u00adene sulfane moiety are orientated almost perpendicular to each other, as indicated by torsion angle \u03c411. Here, either a syn-clinal (68.1\u201388.4\u00b0) or an anti-clinal (92.2\u2013141.1\u00b0) conformation is adopted. However, there is one outlier was synthesized following a well-described literature protocol was dissolved in absolute ethanol (50\u2005ml) under heating and stirring. The resulting solution was slowly added to a hot solution of di-p-meth\u00adoxy\u00adbenzalacetone dissolved in absolute ethanol (50\u2005ml). 3-5 drops of concentrated hydro\u00adchloric acid were added to the mixture, which was subsequently heated and stirred for 5\u2005h : 3145, \u03bd(N\u2014H); 1630, \u03bd(C=N); 1244, \u03bd(N\u2014N); 1059, \u03bd(C=S); 3145. 1H NMR (CDCl3) \u03b4 (p.p.m.)= 6.55\u20137.68 (aromatic H); 3.85 (OCH3); 4.55 (\u2013CH2 benz\u00adyl); 10.20 (N\u2014H). 13C NMR (CDCl3) \u03b4(p.p.m.)= 114.35\u2013142.88 (aromatic C); 55.55 (OCH3); 39.34 (\u2013CH2 benz\u00adyl); 161.22 (C=N); 206.91 (C=S). m/z calculated for C27H26N2O2S2: 474, found 474.Yield: 57.3%. m.p.: 375-376\u2005K. Analysis calculated for CA, B, C or D. The N-bound H atoms were located in difference-Fourier maps and were refined freely [N\u2014H = 0.81\u2005(3)\u20130.91\u2005(4)\u2005\u00c5]. The C-bound H atoms were positioned geometrically (C\u2014H = 0.93\u20130.97\u2005\u00c5) and refined using a riding model, with Uiso(H) = 1.2 or 1.5Ueq(C). A rotating-group model was applied to the methyl groups. The 1,5-bis\u00ad(phen\u00adyl)penta-1,4-dien-3-ylidenyl amine moiety in mol\u00adecule B and the phenyl\u00admethyl moiety in mol\u00adecule D display positional disorder, with refined site occupancy ratios of 0.667\u2005(7):0.333\u2005(7) and 0.653\u2005(15):0.347\u2005(15), respectively .Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989019013458/wm5520sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019013458/wm5520Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019013458/wm5520Isup3.cmlSupporting information file. DOI: 1902915, 1902915CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A third monoclinic polymorph of the title compound is described. In the crystal, it exhibits C\u2014H\u22efCl hydrogen bonds and face-on Cl\u22ef\u03c0 inter\u00adactions involving the chloro\u00adform disolvate mol\u00adecules. Inter\u00admolecular weak offset \u03c0\u2013\u03c0 inter\u00adactions are also present between the aromatic rings of the ligands. 2(C26H22P2)]\u00b72CHCl3 (I), is the third monoclinic polymorph of this platinum(II) complex involving the bidentate ligand cis-1,2-bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)ethyl\u00adene (cis-dppe) , linking the mol\u00adecules to form supra\u00admolecular sheets that lie in the bc plane.The title compound, [PtCll. 1998a. Inorg. al. 1995. Inorg. A selection of recent examples include complexes involving iron(II) (Song cis-dppe ligand and the title compound (I)II metal center with bidentate coordination by the phospho\u00adrus atoms of the cis-dppe ligand. The metal coordination sphere is completed by two chloride anions.The mol\u00adecular structures of the et al., 1998aP21/n without solvent in the unit cell, while structure ZOLYII As for the previously reported polymorphs of compound (I)II center of all three structures are, unsurprisingly, quite similar. The Pt\u2014P bond lengths range from 2.210\u2005(2) to 2.219\u2005(2)\u2005\u00c5, while the Pt\u2014Cl bond lengths range from 2.358\u2005(2) to 2.366\u2005(3)\u2005\u00c5. The P\u2014Pt\u2014P bond angles range from 86.66\u2005(11) to 87.08\u2005(5)\u00b0, while the Cl\u2014Pt\u2014Cl bond angles range from 90.33\u2005(7) to 91.03\u2005(5)\u00b0. The \u03c44 descriptor for fourfold coordination to 3.789\u2005(10)\u2005\u00c5, while the D\u2014H\u22efA bond angles range from 132 to 158\u00b0. Three face-on Cl\u22ef\u03c0 inter\u00adactions nd Fig.\u00a02. The Cl\u22efbc plane, as shown in Fig.\u00a03Cg2\u22efCg2ii = 4.096\u2005(6)\u2005\u00c5 , and Cg3\u22efCg4iii = 3.770\u2005(6)\u2005\u00c5 [Cg3 and Cg4 are the centroids of rings C15\u2013C20 and C21\u2013C26, respectively, \u03b1 = 5.3\u2005(5)\u00b0, inter\u00adplanar distances are 3.326\u2005(4) and 3.439\u2005(4)\u2005\u00c5, slippage = 1.544\u2005\u00c5, symmetry code (iii) \u2212x\u00a0+\u00a01, y\u00a0\u2212\u00a0z\u00a0+\u00a0The complex mol\u00adecules are also linked by weak offset \u03c0\u2013\u03c0 inter\u00adactions, forming sheets that lie in the 2Cl2 solvent mol\u00adecule and one CHCl3 solvent mol\u00adecule in the unit cell, also shows Cl\u22ef\u03c0 inter\u00adactions. However, the methyl\u00adene chloride solvent mol\u00adecule is not engaged in a hydrogen bond with a chlorine atom of the PtII complex, and is disordered in the crystal lattice.The closely related polymorph ZOLYII, which contains one CHet al., 2016cis-dppe ligand is coordinated to a PtII center. In addition to the two polymorphs described above, the most similar cis-dppe\u2013PtII coordination complexes include AFEXEO . Another structure related to the title compound is KADQEL in a NMR tube. This solution was left to stand at room temperature, and colorless needle-like crystals of compound (I)The title compound was prepared serendipitously by mixing 20.5\u2005mg of Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989018008836/su5444sup1.cifCrystal structure: contains datablock(s) Global, I. DOI: 10.1107/S2056989018008836/su5444Isup2.hklStructure factors: contains datablock(s) I. DOI: 1849747CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The synthesis of a hybrid mol\u00adecule is reported. The crystal structure of the monohydrate was investigated using Hirshfeld surface analysis and enrichment contact ratios. Hydrogen bonds induced by guest water mol\u00adecules are the main driving force in crystal packing formation. 14H14N2OS2\u00b7H2O, the planar imidazopyridine ring system is linked to the 1,3-di\u00adthiol\u00adane moiety by an enone bridge. The atoms of the C\u2014C bond in the 1,3-di\u00adthiol\u00adane ring are disordered over two positions with occupancies of 0.579\u2005(14) and 0.421\u2005(14) and both disordered rings adopt a half-chair conformation. The oxygen atom of the enone bridge is involved in a weak intra\u00admolecular C\u2014H\u22efO hydrogen bond, which generates an S(6) graph-set motif. In the crystal, the hybrid mol\u00adecules are associated in R22(14) dimeric units by weak C\u2014H\u22efO inter\u00adactions. O\u2014H\u22efO hydrogen bonds link the water mol\u00adecules, forming infinite self-assembled chains along the b-axis direction to which the dimers are connected via O\u2014H\u22efN hydrogen bonding. Analysis of inter\u00admolecular contacts using Hirshfeld surface analysis and contact enrichment ratio descriptors indicate that hydrogen bonds induced by water mol\u00adecules are the main driving force in the crystal packing formation.In the title hydrated hybrid compound C Its geometrical parameters are similar to those found for 1--3,3-bis\u00ad(methyl\u00adsulfan\u00adyl)prop-2-enone = 0.419\u2005(7)/0.443\u2005(9)\u2005\u00c5, \u03c6(2) = 303.2\u2005(9)/128.9\u2005(11)\u00b0 for the major and minor components, respectively]. The oxygen atom of the linker moiety is involved in a weak intra\u00admolecular C6\u2014H6\u22efO1 hydrogen bond (Table\u00a01S(6) graph-set motif.Fig.\u00a01d Table\u00a01, which gvia pairwise weak C\u2014H\u22efO inter\u00adactions [H5\u22efO1i = 2.71\u2005\u00c5; symmetry code as in Table\u00a01W\u2014H1W\u22efO2Wii hydrogen bonds, generating an infinite self-assembled chain of water mol\u00adecules in a helical fashion along the b axis around which the host mol\u00adecules are linked via O2W\u2014H2W\u22efN1 hydrogen bonds and weak C12\u2014H12D\u22efO2Wii inter\u00adactions .In the crystal, the host mol\u00adecules form inversion dimers ns Fig.\u00a04. The hoson Fig.\u00a05 and eachet al., 2007CrystalExplorer are displayed in Figs. 6W\u2014H2W\u22efN1 hydrogen bond while the pale-red spot near H12B illustrates the weak C\u2014H\u22efO2W inter\u00adaction. The white spots represent H\u22efO, H\u22efS and H\u22efH contacts. On the shape-index surface, convex blue regions indicate hydrogen-donor groups, while concave red regions indicate hydrogen-acceptor groups and S\u22efN and S\u22efC contacts and O\u22efC inter\u00adactions. The fingerprint plots show the contribution of different types of inter\u00admolecular inter\u00adactions is defined as the ratio between the proportion of actual crystal contacts between the different chemical species and the theoretical proportion of random equiprobable contacts . The results obtained are summarized in Table\u00a02ECHc = 0.76 and correspond to weak C\u2014H\u22efC inter\u00adactions. These inter\u00adactions are under-represented because competition with the S\u22efHc, OW\u22efHc and weak O\u22efHc hydrogen bonds, the first two of which appear favoured with enrichment values of 1.35 and 1.14, respectively, and the last slightly under-represented with an enrichment ratio of 0.98. The C\u22efC contacts are privileged and display an enrichment value of 1.85, which highlight mol\u00adecules stacking one on top of the other as shown in Fig.\u00a05et al. -3,3-bis\u00ad(methyl\u00adsulfan\u00adyl)prop-2-\u220anone monohydrate ethanone (6.2\u2005mmol) was dissolved in distilled dimethyl sulfoxide (15\u2005ml), and the carbon di\u00adsulfide was added. After cooling the mixture to 273\u2005K, sodium hydride was added. After stirring for 30 min. at 273\u2005K, the mixture was stirred at ambient temperature for 4\u2005h. The solution was then cooled at 273\u2005K and 1,2-di\u00adchloro ethane was added dropwise. The resulting mixture was then stirred for 24\u2005h and then poured into 50\u2005ml of ice-cold water. The precipitate was filtered and recrystallized from a mixture of water\u2013dioxane (2:1) to obtain brown single crystals of the title compound suitable for X-ray diffraction analysis .1- and were refined using a riding model with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-meth\u00adyl). In the 1,3-di\u00adthiol\u00adane ring, the carbon atoms of the C\u2014C bond are disordered over two positions with refined occupancy factors of 0.579\u2005(14) and 0.421\u2005(14). C\u2014C bond lengths in both disordered components were restrained to the target value of 1.513\u2005\u00c5 I. DOI: Click here for additional data file.10.1107/S2056989019015755/vm2224Isup2.cmlSupporting information file. DOI: 1967239CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "PTPN2 deletion in T cells enhances cancer immunosurveillance and the efficacy of adoptively transferred tumour\u2010specific T cells. T\u2010cell\u2010specific PTPN2 deficiency prevented tumours forming in aged mice heterozygous for the tumour suppressor p53. Adoptive transfer of PTPN2\u2010deficient CD8+ T cells markedly repressed tumour formation in mice bearing mammary tumours. Moreover, PTPN2 deletion in T cells expressing a chimeric antigen receptor (CAR) specific for the oncoprotein HER\u20102 increased the activation of the Src family kinase LCK and cytokine\u2010induced STAT\u20105 signalling, thereby enhancing both CAR T\u2010cell activation and homing to CXCL9/10\u2010expressing tumours to eradicate HER\u20102+ mammary tumours in\u00a0vivo. Our findings define PTPN2 as a target for bolstering T\u2010cell\u2010mediated anti\u2010tumour immunity and CAR T\u2010cell therapy against solid tumours.Although adoptive T\u2010cell therapy has shown remarkable clinical efficacy in haematological malignancies, its success in combating solid tumours has been limited. Here, we report that Protein tyrosine phosphatase N2 inhibits tumour T\u2010cell infiltration and CAR T\u2010cell cytotoxicity and may thus be a therapeutic target to enhance solid\u2010tumour immunotherapy. Upon necropsy 15/28 (54%), Ptpn2fl/fl;p53+/\u2212 mice developed various tumours including thymomas, lymphomas, sarcomas, carcinomas and hepatomas were evident in the thymi or peripheral lymphoid organs of 5/28 mice developed any overt tumours, splenomegaly or abnormal lymphocytic populations as assessed by gross morphology or flow cytometry and lymphoid organ tissue architecture was normal \u2010expressing AT\u20103 (AT\u20103\u2010OVA) mammary carcinoma cells implanted into the inguinal mammary fat pads of Ptpn2fl/fl versus Lck\u2010Cre;Ptpn2fl/fl C57BL/6 mice T cells into tumours were increased rather than decreased in AT\u20103\u2010OVA tumours \u2010\u03b3 and tumour necrosis factor (TNF) peptide SIINFEKL. Naive OT\u20101 T cells can undergo clonal expansion and develop effector function when they engage OVA\u2010expressing tumours, but thereon leave the tumour microenvironment, become tolerised and fail to control tumour growth Ptpn2fl/fl OT\u20101 CD8+ T cells had no overt effect on the growth of AT\u20103\u2010OVA mammary tumours when compared to vehicle\u2010treated tumour\u2010bearing mice by 25\u00a0days. Tumour re\u2010emergence in this setting was accompanied by decreased OVA and MHC class I (H2\u2010k1) gene expression, consistent with decreased antigen presentation; tumour re\u2010emergence was also accompanied by decreased PD\u2010L1 (Cd274) gene expression cells that contribute to immunosurveillance and the suppression of solid tumour formation subset sarcoma cells, but importantly, not HER\u20102\u2010negative 24JK control cells and central memory (CD44hiCD62Lhi) PTPN2\u2010deficient CD8+ HER\u20102 CAR T cells were more effective at specifically killing 24JK\u2010HER\u20102 cells but not 24JK cells in\u00a0vitro of purified Ptpn2fl/fl versus Lck\u2010Cre;Ptpn2fl/fl central memory CD8+ HER\u20102 CAR T cells into sub\u2010lethally irradiated syngeneic recipients bearing established orthotopic tumours arising from the injection of HER\u20102\u2010expressing E0771 (HER\u20102\u2010E0771) mammary tumour cells and IL\u201010 (Il10) of the maximal ethically permissible mammary tumour burden prior to CAR T\u2010cell therapy rather than the mixed central and effector/memory (CD44hiCD62Llo) phenotypes otherwise present on day 10 post\u2010adoptive transfer . When CD8+ T cells are activated by a strong TCR stimulus and subsequently stimulated with IL\u20102, they undergo differentiation into effectors and acquire CTL activity characterised by IFN\u03b3 and granzyme B expression . Since IL\u20102\u2010induced STAT\u20105 signalling was only partially corrected in Lck\u2010Cre;Ptpn2fl/fl;Lck+/\u2212 CAR T cells being reduced to those in Ptpn2fl/fl control CAR T cells Fig\u00a0B. To asset\u00a0al, et\u00a0al, Ptpn2 in murine CAR T cells by RNA interference using nuclease resistant siRNA duplexes (siSTABLE\u2122) that efficiently knockdown genes for prolonged periods Fig\u00a0F. By conet\u00a0al, ex\u00a0vivo so that resultant CAR T cells do not overexpress Cas9 and do not elicit immunogenic responses post\u2010adoptive transfer. To this end, we transfected total CAR T cells with recombinant nuclear\u2010localised Cas9 pre\u2010complexed with short guide (sg) RNAs capable of directing Cas9 to the Ptpn2 locus and the human melanoma cell line MDA\u2010MB\u2010435 (ATCC\u00aeHTB\u2010219\u2122) were obtained from the ATCC, Manassas, Virginia, USA. HER\u20102\u2010E0771 cells were engineered to inducibly overexpress murine PTPN2 (HER\u20102\u2010E0771\u2010PTPN2hi) in response to doxycycline using the Tet\u2010On 3G Inducible Expression System according to the manufacturer's instructions (Clontech). Tumour cells were cultured in RPMI 1640 or high\u2010glucose DMEM (AT3\u2010OVA) supplemented with 10% FBS, l\u2010glutamine (2\u00a0mM), penicillin (100 units/ml)/streptomycin (100\u00a0\u03bcg/ml), MEM non\u2010essential amino acids (0.1\u00a0mM), sodium\u2010pyruvate (1\u00a0mM), HEPES (10\u00a0mM) and 2\u2010mercaptoethanol (50\u00a0\u03bcM).The C57BL/6 mouse mammary carcinoma cell line E0771 (Johnstone PtprcaPepcb/BoyJ) and human HER\u20102 transgenic (TG) recipient mice and 6\u2010 to 8\u2010week\u2010old female donor mice were used for adoptive transfers. For ex\u00a0vivo experiments, either male or female mice were used. Aged\u2010 and sex\u2010matched littermates were used in all experiments. Ptpn2fl/fl and Lck\u2010Cre;Ptpn2fl/fl mice and the corresponding OT\u20101 TCR transgenic mice were described previously mice have been described previously with Lck\u2010Cre;Ptpn2fl/fl mice. B6.129S2\u2010Lck/J mice were a gift from Dr Andre Veillette and were bred with Lck\u2010Cre;Ptpn2fl/fl mice to generate Lck\u2010Cre;Ptpn2fl/fl;Lck+/\u2212 mice. Ly5.1 and C57BL/6 mice were purchased from the WEHI Animal Facility , and human HER\u20102 (C57BL/6) transgenic (TG) mice were bred and maintained at the Peter MacCallum Cancer Centre.Mice were maintained on a 12\u2010h light\u2013dark cycle in a temperature\u2010controlled high barrier facility with free access to food and water. Six\u2010 to 10\u2010week\u2010old female Ly5.1 was purchased from Thermo Fisher Scientific. The mouse antibody against PTPN2 (clone 6F3) was provided by M. Tremblay (McGill University). For immunofluorescence staining and immunohistochemistry, rabbit anti\u2010CD3\u03b5 and mouse anti\u2010HER\u20102 from Abcam were used. Recombinant human IL\u20102, murine IL\u20107 and IL\u201015 used for T\u2010cell stimulation or IFN\u03b3 used for stimulating tumour cells were purchased from the NIH or PeproTech, respectively. RetroNectin was purchased from Takara, Dnase I and doxycycline from Sigma\u2010Aldrich. The mouse anti\u2010nuclear antibodies Ig's ELISA Kit (Alpha Diagnostic Int.) and the Transaminase II Kit were used according to the manufacturer's instructions. FBS was purchased from Thermo Scientific; Dulbecco\u2010Phosphate\u2010Buffered Saline (D\u2010PBS), RPMI 1640, DMEM, MEM non\u2010essential amino acids and sodium\u2010pyruvate were from Invitrogen, and collagenase type IV was purchased from Worthington Biochemical.For cell stimulation, anti\u2010CD3\u03b5 (clone 145\u20102C11) and anti\u2010CD28 (clone 37.51) antibodies were purchased from BD Biosciences. Antibodies against p\u2010(Y701) STAT1 (clone 58D6), p\u2010(Y694) STAT5 (D47E7) XPet\u00a0al, et\u00a0al, Single\u2010cell suspensions from spleen, lymph nodes and hepatic lymphocytes were obtained as previously described or CyAn\u2122 ADP (Beckman Coulter).For FACS sorting, cells were stained in 15\u2010ml Falcon tubes (BD Biosciences) for 30\u00a0min on ice and purified using either a BD Influx cell sorter, or the BD FACSAria II, BD FACSAria Fusion 3 or BD FACSAria Fusion 5 instruments.Data were analysed using FlowJo8.7 or FlowJo10 (Tree Star Inc.) software. For cell quantification, a known number of Calibrite\u2122 Beads (BD Biosciences) or Nile Red Beads (ProsiTech) or Flow\u2010Count Fluorospheres (Beckman Coulter) were added to samples before analysis.The following antibodies from BD Biosciences, BioLegend or eBioscience were used for flow cytometry: Phycoerythrin (PE) or peridinin\u2013chlorophyll cyanine 5.5 (PerCP\u2010Cy5.5)\u2010conjugated CD3 (145\u20102C11); PerCP\u2010Cy5.5 or phycoerythrin\u2013cyanine 7 (PE\u2010Cy7)\u2010conjugated CD4 (RM4\u20105); BV711, Pacific Blue\u2010conjugated (PB), allophycocyanin (APC)\u2010Cy7 or APC\u2010conjugated CD8 (53\u20106.7); PerCP\u2010Cy5.5, PE or APC\u2010Cy7\u2010conjugated CD25 (P61); Fluorescein isothiocyanate (FITC), V450, Alexa Fluor 700 or PE\u2010Cy7\u2010conjugated CD44 (IM7); APC\u2010conjugated CD45 (30\u2010F11); V450, APC, Alexa Fluor 488 or PE\u2010conjugated CD45.1 ; FITC, PB or PerCP\u2010Cy5.5\u2010conjugated CD45.2 ; APC\u2010conjugated CD45R ; PE\u2010Cy7, APC or PE\u2010conjugated CD62L (Mel\u201014); PE\u2010conjugated CD122 (TM\u2010\u03b21); PE\u2010conjugated CD132 (4G3); Alexa Fluor 647\u2010conjugated CD183 (CXCR3\u2010173); PerCP\u2010eFluo710\u2010conjugated CD185 ; PE\u2010Cy7 conjugated CD197 ; PE\u2010conjugated CD223 ; PE\u2010Cy7\u2010conjugated CD279 ; FITC\u2010conjugated CD11b (M1/70); APC\u2010Cy7\u2010conjugated Ly6C (AL\u201021); PE\u2010conjugated Ly\u20106G (clone 1A8); APC\u2010conjugated F4/80 (BM8); PE\u2010conjugated IL\u201010 (JES5\u201016E3), PE\u2010conjugated Nur77 (12.14); PE\u2010Cy7\u2010conjugated IFN\u03b3 (XMG1.2); FITC or APC\u2010conjugated TNF (MP6\u2010XT22); Alexa Fluor 647\u2010conjugated Granzyme B (GB11); PE\u2010Cy7\u2010conjugated TCR\u2010V\u03b12 (B20.1); eFluor 660\u2010conjugated p(Y418)\u2010Src (SC1T2M3); Alexa Fluor 647\u2010conjugated T\u2010bet (4B10); and V450\u2010conjugated FoxP3 (clone MF23).The following antibodies from Miltenyi Biotec were used for flow cytometry: PE\u2010conjugated CD197 ; VioBright FITC\u2010conjugated CD8 (BW135/80); and VioBlue\u2010conjugated CD45RA (REA1047).7/ml) were cultured overnight with anti\u2010CD3\u03b5 (0.5\u00a0\u03bcg/ml) and anti\u2010CD28 (0.5\u00a0\u03bcg/ml) in the presence of 100\u00a0IU/ml human IL\u20102 and 0.2\u00a0ng/ml murine IL\u20107 in complete T\u2010cell medium . Dead cells were removed by Ficoll centrifugation according to the manufacturer's instructions. Retrovirus encoding a second\u2010generation chimeric antibody receptor (CAR) consisting of an extracellular scFv\u2010anti\u2010human HER\u20102, a membrane proximal CD8 hinge region and the transmembrane and the cytoplasmic signalling domains of CD28 fused to the cytoplasmic region of CD3\u03b6 (scFv\u2010anti\u2010HER\u20102\u2010CD28\u2010\u03b6) was obtained from the supernatant of the GP+E86 packaging line as described previously at room temperature and incubated overnight before the second viral transduction. T cells were maintained in IL\u20102\u2010 and IL\u20107\u2010containing media and cells used at days 7\u20138 after transduction.Splenocytes were isolated from murine spleens by mechanical disruption, and contaminating red blood cells were removed by incubation with 1\u00a0ml of Blood Cell Lysing Buffer Hybri\u2010Max\u2122 (Sigma\u2010Aldrich) for 7\u00a0min at room temperature per spleen. Lymphocytes CAR T cells and expanded for up to 15\u00a0days in the presence of human IL\u20102. The expression of the chimeric receptor was determined by staining with Alexa Flour 647\u2010conjugated anti\u20103S193 idiotype [supplied by Ludwig Institute for Cancer Research (LICR)], and truncated CD34 was detected with PE\u2010conjugated anti\u2010human CD34 (BioLegend). Cell surface phenotyping of transduced cells was determined by staining with BV785\u2010conjugated anti\u2010human CD3 , BV605\u2010conjugated anti\u2010human CD4 and PE\u2010Cy7\u2010conjugated anti\u2010human CD8 .Human peripheral blood mononuclear cells (PBMCs) were isolated from normal donor buffy coats. PBMCs were stimulated with anti\u2010human CD3 and human IL\u20102 (600\u00a0IU/ml) in complete T\u2010cell medium. Retrovirus encoding a second\u2010generation chimeric antibody receptor (CAR) consisting of an extracellular scFv\u2010anti\u2010human LeY domain, a membrane proximal CD8 hinge region and the transmembrane and the cytoplasmic signalling domains of CD28 fused to the cytoplasmic region of CD3\u03b6 was obtained from the supernatant of the PG13 packaging cell line and has been described previously (Westwood 5) generated from human PBMC were incubated with Lewis Y\u2010expressing OVCAR\u20103 cells (1\u00a0\u00d7\u00a0105) or Lewis Y\u2010negative MDA\u2010MB\u2010435 cells (1\u00a0\u00d7\u00a0105) or plate\u2010bound anti\u2010LY for 6\u00a0h at 37\u00b0C in complete T\u2010cell medium. GolgiPlug\u2122 and GolgiStop\u2122 were added 3\u00a0h before cells were processed for surface and intracellular staining for CD4 , CD8 , IFN\u03b3 and TNF by flow cytometry.CAR T cells or IL\u201015 (5 or 10\u00a0ng/ml) in 100\u00a0\u03bcl RPMI/1 %FBS. Cells were processed for intracellular p(Y694)\u2010STAT\u20105 detection by flow cytometry as previously described dilution, purified CD8+CD44hiCD62Lhi CAR T cells were incubated with CTV in D\u2010PBS supplemented with 0.1% (v/v) BSA at a final concentration of 2\u00a0\u03bcM for 10\u00a0min at 37\u00b0C. Cells were then washed three times with D\u2010PBS supplemented with 10% (v/v) FBS. CAR T cells (2\u00a0\u00d7\u00a0105) were incubated with HER\u20102\u2010expressing or HER\u20102\u2010negative 24JK sarcoma cells (1\u00a0\u00d7\u00a0105) in complete T\u2010cell medium.CAR T cells generated from mouse splenocytes were stained with fluorochrome\u2010conjugated antibodies for CD8, CD62L and CD44 and sorted for CD8+CD44hiCD62Lhi CAR T cells were incubated with plate\u2010bound anti\u2010CD3\u03b5 for 24\u00a0h at 37\u00b0C in complete T\u2010cell medium. Pre\u2010activated CAR T Cells (2\u00a0\u00d7\u00a0105) were labelled with 5\u00a0\u03bcM CTV and incubated with HER\u20102\u2010expressing or HER\u20102\u2010negative 24JK sarcoma cells (1\u00a0\u00d7\u00a0105) in complete T\u2010cell medium. At various time points, proliferating cells were harvested and CTV dilution was monitored by flow cytometry. For cell quantification, Nile Red Beads (ProsiTech) or Flow\u2010Count Fluorospheres (Beckman Coulter) were added to samples before analysis.For the assessment of CAR T\u2010cell proliferation after anti\u2010CD3\u03b5 stimulation, purified CD8T cells were incubated rocking on an orbital shaker with PTPN2\u2010inhibitor compound 8 (100\u00a0nM) or vehicle (DMSO 1% v/v) in serum\u2010free RPMI 1640 for 1\u00a0h at 37\u00b0C. Cells were washed three times with complete T\u2010cell medium and processed for T\u2010cell activation studies.5 HER\u20102\u2010E0771 cells resuspended in 20\u00a0\u03bcl D\u2010PBS into the fourth mammary fat pad. At day 6 post\u2010tumour cell injection, human HER\u20102 TG mice were pre\u2010conditioned with total body irradiation (4\u00a0Gy) prior to the adoptive transfer of 6\u00a0\u00d7\u00a0106 FACS\u2010purified CD8+CD44hiCD62Lhi CAR T cells. Mice were treated with 50,000\u00a0IU IL\u20102 on days 0\u20134 after T\u2010cell transfer.Female human HER\u20102 TG mice were anaesthetised with Ketamine (100\u00a0mg/kg) and Xylazil (10\u00a0mg/kg) (Troy Laboratories) and injected orthotopically with 2\u00a0\u00d7\u00a0102) was abraded using a MultiPro Dremel with a grindstone attachment. B16.F10\u2010OVA\u2010expressing melanoma cells (1\u00a0\u00d7\u00a0105) were resuspended in 10\u00a0\u03bcl of Matrigel\u2122 (BD Biosciences) and orthotopically applied to the lesion. Set Matrigel was covered with a piece of Op\u2010site FlexigridTM (Smith and Nephew). The torsos of the mice were wrapped with a soft hypoallergenic MicroporeTM tape and then by a stronger porous polyethylene TransporeTM tape to protect the abraded site. Twenty\u2010four hours later, 2\u00a0\u00d7\u00a0105 FACS\u2010purified CD8+ naive CD44loCD62Lhi OT\u20101 T cells were adoptively transferred. Recipient mice were monitored daily until bandages were removed 6\u00a0days after tumour engraftment.Ly5.1 mice were anaesthetised with Ketamine (100\u00a0mg/kg) and Xylazil (10\u00a0mg/kg) (Troy Laboratories). The flanks of the mice were shaved using clippers and depilated with Veet (Reckitt Benckiser), and the skin and Xylazil (10\u00a0mg/kg) (Troy Laboratories) and were injected orthotopically with 1\u00a0\u00d7\u00a0106/200\u00a0\u03bcl) were stimulated with PMA and ionomycin in the presence of GolgiPlug\u2122 and GolgiStop\u2122 (BD Biosciences) for 4\u00a0h at 37\u00b0C in complete T\u2010cell medium.Tumour\u2010bearing mice were sacrificed, and tumours were excised and digested at 37\u00b0C for 30\u00a0min using a cocktail of 1\u00a0mg/ml collagenase type IV and 0.02\u00a0mg/ml DNase (Sigma\u2010Aldrich) in DMEM supplemented with 2% (v/v) FBS. Cells were passed through a 70\u2010\u03bcm cell strainer (BD Biosciences) twice and processed for flow cytometry. For the detection of intracellular cytokines in tumour\u2010infiltrating lymphocytes, cells and 10\u00a0mM HEPES for 1\u00a0h at 37\u00b0C. Cells were passed through a 70\u2010\u03bcm cell strainer twice and washed three times with 50\u00a0ml PBS. Cells were resuspended in DMEM supplemented with 10% FBS, 2\u00a0mM l\u2010glutamine, penicillin (100 units/ml), streptomycin (100\u00a0\u03bcg/ml) and seeded into 24\u2010well plates. Cells were incubated at 37\u00b0C until they reached 80% confluency. Tumour\u2010infiltrating T cells from AT\u20103\u2010OVA tumour\u2010bearing C57BL/6 mice Ptpn2fl/fl and Lck\u2010Cre;Ptpn2fl/fl mice were labelled with 2\u00a0\u03bcM CTV and incubated with adherent, 80% confluent AT\u20103\u2010OVA tumour cells overnight. GolgiPlug\u2122 and GolgiStop\u2122 were added 3\u00a0h before cells were processed for intracellular staining for IFN\u03b3 and TNF by flow cytometry.AT\u20103\u2010OVA tumour\u2010bearing C57BL/6 mice were sacrificed, and tumours were collected in ice\u2010cold D\u2010PBS and then digested with 50\u00a0\u03bcg/ml Liberase in Hank's Balanced Salt Solution or effector/memory (CD44hiCD62Llo) or total HER\u20102\u2010specific CAR T cells were added at different concentrations to the mix of HER\u20102\u2010expressing (5\u00a0\u00d7\u00a0104) and HER\u20102\u2010negative (5\u00a0\u00d7\u00a0104) 24JK sarcoma cells and incubated for 4\u00a0h at 37\u00b0C in complete T\u2010cell medium. Antigen\u2010specific target cell lysis (24JK\u2010HER\u20102 cell depletion) was monitored by flow cytometry.For the assessment of CAR T\u2010cell cytotoxicity, HER\u20102\u2010expressing 24JK sarcoma cells (5\u00a0\u03bcM CTV) and HER\u20102\u2010negative 24JK sarcoma cells (0.5\u00a0\u03bcM CTV) were labelled with CellTrace\u2122 Violet in D\u2010PBS supplemented with 0.1% (v/v) BSA for 15\u00a0min at 37\u00b0C. Tumour cells were then washed three times with D\u2010PBS supplemented with 10% (v/v) FBS and mixed at a 1:1 ratio. FACS\u2010purified CD8For brain immunohistochemistry, mice were perfused transcardially with heparinised saline followed by 4% (w/v) paraformaldehyde in phosphate buffer . Brains were post\u2010fixed overnight and then kept for 4\u00a0days in 30% (w/v) sucrose in 0.1\u00a0M phosphate buffer to cryoprotect the tissue, before freezing on dry ice. 30\u2010\u03bcm sections (120\u00a0\u03bcm apart) were cut in the coronal plane throughout the entire rostral\u2010caudal extent of the cerebellum. For detection of CD3\u03b5, sections were subjected to antigen retrieval in citrate acid buffer at 85\u00b0C for 20\u00a0min. Sections were incubated at room temperature for 2\u00a0h in blocking buffer [0.1M phosphate buffer, 0.2% (v/v) Triton X\u2010100, 10% (v/v) normal goat serum (Sigma\u2010Aldrich) and then overnight at 4\u00b0C in rabbit anti\u2010CD3 (Dako) in 1% (v/v) blocking buffer. After washing with PBS, sections were incubated with goat anti\u2010rabbit Alexa Fluor 488\u2010conjugated secondary antibody (Life Technologies) in blocking buffer for 2\u00a0h at room temperature. Sections were mounted with Mowiol 4\u201088 mounting media and visualised using an Olympus Provis AX70 microscope. Images were captured with an Olympus DP70 digital camera and processed using AnalySIS (Olympus) software. To assess gross tissue morphology, a subset of sections throughout the entire rostral\u2010caudal extent of the cerebellum were stained with haematoxylin and eosin.For detection of HER\u20102, 30\u2010\u03bcm sections throughout the entire rostral\u2010caudal extent of the cerebellum from C57BL/6 or HER\u20102 TG C57BL/6 mice were subjected to antigen retrieval and incubated in blocking buffer (as described above). Sections were then incubated overnight in (4\u00b0C) with anti\u2010c\u2010ErbB2/c\u2010Neu (Ab\u20103) mouse mAb (3B5) . After washing with PBS, HER\u20102\u2010positive cells were visualised using rabbit IgG VECTORSTAIN ABC Elite and DAB Peroxidase Substrate Kits and visualised using a bright field microscope.For mammary fat pad tumour immunohistochemistry, animals were culled and mammary fat pad immediately dissected and fixed in buffered formalin solution for 48\u00a0h. Tissues were embedded in paraffin and 4\u2010\u03bcm sections of the entire block prepared. Every tenth to fourteenth section of the tissue was used to detect HER\u20102 and CD3 by immunohistochemistry. After deparaffinisation and rehydration, sections were subjected to antigen retrieval in Tris/EDTA buffer (pH 8.0) at 120\u00b0C for 10\u00a0min. Sections were blocked with 5% (v/v) bovine serum albumin in 0.1\u00a0M phosphate buffer for 1\u00a0h at room temperature and incubated overnight (4\u00b0C) with anti\u2010c\u2010ErbB2/c\u2010Neu (Ab\u20103) mouse mAb (3B5) or anti\u2010CD3 . After washing with PBS, HER\u20102\u2010 and CD3\u2010positive cells were visualised using rabbit IgG VECTORSTAIN ABC Elite and DAB Peroxidase Substrate Kits and counterstained with haematoxylin. Sections were visualised on a Zeiss Axioskop 2 mot plus microscope .\u2122 SYBR\u00ae Green Assay were utilised to perform quantitative PCR detecting Cxcl9, Cxcl10, Tgfb1, Cd274, H2\u2010K1 and Rps18. Primer sets for HER\u20102, Nono and SerpinB14 were purchased from Sigma\u2010Aldrich. Relative gene expression (\u0394Ct) was determined by normalisation to the house\u2010keeping gene Nono throughout the study except for Fig\u00a0Rps18 was used and \u0394\u0394Ct analysis performed.RNA was extracted with TRIzol reagent and RNA quality and quantity determined using a NanoDrop 2000 (Thermo Fisher Scientific). mRNA was reverse transcribed using a High\u2010Capacity cDNA Reverse Transcription Kit and processed for quantitative real\u2010time PCR either using the Fast SYBR\u2122 Green Master Mix . Primer sets from PrimePCRPtpn2 was knocked down transiently in HER\u20102 CAR T cells using nuclease resistant Ptpn2 siRNA duplexes ; siRNA duplexes (siSTABLE\u2122) for enhanced green fluorescent protein were used as a control. CAR T cells were transfected with Ptpn2 siRNA (300\u00a0nM) conjugated to FITC or GFP siRNA (300\u00a0nM) conjugated to FITC 2\u00a0days prior to adoptive T\u2010cell therapy using the Mouse T\u2010cell Nucleofector\u2122 Kit (Lonza Bioscience) according to the manufacturer's instructions.Ptpn2 was deleted in HER\u20102 CAR T cells using Cas9 ribonucleoprotein (RNP)\u2010mediated gene editing. Briefly, total CAR T cells were transfected with recombinant Cas9 pre\u2010complexed with short guide (sg) RNAs targeting the Ptpn2 locus or non\u2010targeting sgRNAs (GCACUACCAG AGCUAA CUCA) as a control 2\u00a0days prior to adoptive T\u2010cell therapy using the P3 Primary Cell 4D\u2010Nucleofector X\u2122 Kit (Lonza Bioscience) according to the manufacturer's instructions.hi cell line. Briefly, murine Ptpn2 cDNA from HER\u20102\u2010E0771 cells was reverse transcribed and amplified by PCR and cloned into the SmaI and EcoR1 restriction sites of Tre\u20103G plasmid (Clontech Laboratories) to generate the Tre\u20103G\u2010Ptpn2 construct; the fidelity of the cloned cDNA was confirmed by dideoxy sequencing. HEK293T cells were transfected with either the Tre\u20103G\u2010Ptpn2 or Tet\u2010On 3G constructs using the Lenti\u2010X Packing system (Clontech Laboratories) according to the manufacturer's instructions. HER\u20102\u2010E0771 mammary tumour cells were transduced first with Tet\u2010On 3G lentivirus and selected with G418 (0.8\u00a0mg/ml) and subsequently transduced with Tre\u20103G\u2010Ptpn2 lentivirus and selected with puromycin (1.5\u00a0\u03bcg/ml). Where indicated, the resultant HER\u20102\u2010E0771\u2010PTPN2hi cells were incubated with 2\u00a0mg/ml doxycycline (DOX) to induce the expression of PTPN2. Cells were serum starved in DMEM medium without FBS for 12\u00a0h and stimulated with 2\u00a0ng/ml IFN\u03b3 for the indicated times and processed for immunoblotting or incubated with 1\u00a0ng/ml IFN\u03b3 for 24\u00a0h and processed for quantitative real\u2010time PCR.The Tet\u2010on 3G inducible expression system (Clontech Laboratories) was used to generate the HER\u20102\u2010E0771\u2010PTPN2in\u00a0vivo studies, 2\u00a0\u00d7\u00a0105 HER\u20102\u2010E0771\u2010PTPN2hi cells were resuspended in 20\u00a0\u03bcl D\u2010PBS and injected orthotopically into the fourth mammary fat pad of female human HER\u20102 TG mice. At day 5 post\u2010tumour cell injection, mice were administered DOX (2\u00a0mg/ml) in the drinking water for the entirety of the experiment. At day 6 post\u2010tumour cell injection, human HER\u20102 TG mice were pre\u2010conditioned with total body irradiation (4\u00a0Gy) prior to the adoptive transfer of 6\u00a0\u00d7\u00a0106 FACS\u2010purified CD8+CD44hiCD62Lhi CAR T cells. Mice were treated with 50,000\u00a0IU IL\u20102 on days 0\u20134 after T\u2010cell transfer.For U\u2010test, the parametric 2\u2010tailed Student's t\u2010test, the 1\u2010way or 2\u2010way ANOVA test using Turkey or Sidak post hoc comparison or the log\u2010rank (Mantel\u2013Cox test) where indicated. *P\u00a0<\u00a00.05, **P\u00a0<\u00a00.01, ***P\u00a0<\u00a00.001 and ****P\u00a0<\u00a00.0001 were considered as significant.Statistical analyses were performed with GraphPad Prism software 7.0b using the non\u2010parametric using 2\u2010tailed Mann\u2013Whitney All experiments were performed in accordance with the NHMRC Australian Code of Practice for the Care and Use of Animals. All protocols were approved by the Monash University School of Biomedical Sciences Animal Ethics Committee (Ethics number: MARP/2012/124) or the Peter MacCallum Animal Ethics and Experimentation Committee .Conceptualisation: TT; Methodology: FW, K\u2010HL, XD, SL, KH, GTD, PKG, CK, DM, PAB, MAH, SLP, JW, SZ, Z\u2010YZ, JO, TG, PKD and TT; Data acquisition: FW, K\u2010HL, XD, SL, KH, GTD, PKG, MAH and SLP; Data analysis: FW, K\u2010HL, XD, SL, KH, GTD, PKG; MAH; TG, PKD and TT; Data interpretation: FW, K\u2010HL, XD, SL, KH, GTD, PKG, CK, DM, PAB, MAH, SLP, JW, SZ, Z\u2010YZ, JO, TG, PKD and TT; Writing\u2010Original Draft: TT; Writing\u2010Review and Editing: FW, K\u2010HL, XD, SL, KH, GTD, PKG, CK, DM, PAB, MAH, SLP, JW, SZ, Z\u2010YZ, JO, TG, PKD and TT. Funding acquisition: FW, PKD, TG and TT; Final approval for publishing: FW, K\u2010HL, XD, SL, KH, GTD, PKG, CK, DM, PAB, MAH, SLP, JW, SZ, Z\u2010YZ, JO, TG, PKD and TT.The authors declare that they have no conflict of interest.AppendixClick here for additional data file.Expanded View Figures PDFClick here for additional data file.Source Data for AppendixClick here for additional data file.Review Process FileClick here for additional data file."} +{"text": "The title compound, 2-(4-nitro\u00adphen\u00adyl)-2-oxoethyl 2-chloro\u00adbenzoate, is relatively planar with the two aromatic rings being inclined to each other by 3.56\u2005(11)\u00b0. 15H10ClNO5, is relatively planar with the two aromatic rings being inclined to each other by 3.56\u2005(11)\u00b0. The central \u2014C(=O)\u2014C\u2013O\u2014C(=O)\u2014 bridge is slightly twisted, with a C\u2014C\u2014O\u2014C torsion angle of 164.95\u2005(16)\u00b0. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO and C\u2014H\u22efCl hydrogen bonds, forming layers parallel to the (101) plane. The layers are linked by a further C\u2014H\u22efO hydrogen bond, forming a three-dimensional supra\u00admolecular structure. There are a number of offset \u03c0\u2013\u03c0 inter\u00adactions present between the layers [inter\u00adcentroid distances vary from 3.8264\u2005(15) to 3.9775\u2005(14)\u2005\u00c5]. Hirshfeld surface analyses, the dnorm surfaces, electrostatic potential and two-dimensional fingerprint plots were examined to verify the contributions of the different inter\u00admolecular contacts within the supra\u00admolecular structure. The shape-index surface shows that two sides of the mol\u00adecule are involved in the same contacts with neighbouring mol\u00adecules, and the curvedness plot shows flat surface patches that are characteristic of planar stacking.The title compound, C Torsion angle \u03c41 at \u221212.5\u2005(3)\u00b0 signifies a certain noncoplanarity between the benzene ring (C10\u2013C15) and the adjacent carbonyl group (C9=O3) as a result of steric repulsion between the substituent Cl1 and the adjacent carbonyl group C9=O3. This is also reflected in the torsion angle \u03c42 of \u2212164.95\u2005(16)\u00b0, between the two carbonyl groups, C7=O2 and C9=O3, which have a \u2013anti\u00adperiplanar conformation. Torsion angle \u03c43, involving the benzene ring (C1\u2013C6) and the adjacent carbonyl group (C7=O2), is \u22123.6\u2005(3)\u00b0 and indicates a \u2013synperiplanar conformation. A comparison of the torsion angles in I and II, indicates that the insertion of the Cl atom in I has the most significant influence on torsion \u03c42, which is \u2212164.95\u2005(16)\u00b0 in I compared to 174.08\u2005(9)\u00b0 in II. Torsion angles \u03c41 of \u221212.5\u2005(3)\u00b0 and \u03c43 of \u22123.6\u2005(3)\u00b0 are slightly larger than the values observed in II, viz. 9.60\u2005(16) and 1.88\u2005(15)\u00b0, respectively. Hence, compound I has a less planar conformation than unsubstituted compound II.The overall mol\u00adecular conformation of I is characterized by three torsion angles, i, C14\u2014H14\u22efO4i and C13\u2014H13\u22efCl1iii hydrogen bonds to form layers lying parallel to the (101) plane; see Fig.\u00a02A\u22efO3ii hydrogen bonds and offset \u03c0\u2013\u03c0 inter\u00adactions shows the Hirshfeld surface mapped over dnorm (\u22120.154 to 1.305) and for Fig.\u00a04b) the electrostatic potential. The Hirshfeld surface illustrated in Fig.\u00a04a) reflects the involvement of different atoms with the inter\u00admolecular inter\u00adactions through the appearance of blue and red patches, which correspond to the regions of positive and negative electrostatic potential shown in Fig.\u00a04b). The shape-index surface clearly shows that the two sides of the mol\u00adecule are involved in contacts with neighbouring mol\u00adecules and the curvedness plot shows flat surface patches characteristic of planar stacking.The Hirshfeld surface analysis -2-oxoethyl 4-nitro\u00adbenzoate \u00b0 in the title compound. In the crystal structure of the recently published compound 2-(4-nitro\u00adphen\u00adyl)-2-oxoethyl benzoate (II) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019014336/su5521sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019014336/su5521Isup2.hklStructure factors: contains datablock(s) I. DOI: 1449647, 1449647CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, phenyl-C\u2014H\u22efO(carbon\u00adyl) and carbonyl-O\u22efO(carbon\u00adyl) inter\u00adactions general a [111] supra\u00admolecular chain.In Ru 3(C19H17PS)(CO)11], comprises a triangle of Ru0 atoms, two of which are bonded to four carbonyl ligands. The third metal atom is bound to three carbonyl ligands and the phosphane-P atom of a dissymmetric phosphane ligand, PPh2(C6H4SMe-4); no Ru\u22efS inter\u00adactions are observed. The phosphane occupies an equatorial position and its proximity to an Ru\u2014Ru edge results in the elongation of this bond with respect to the others [2.8933\u2005(2)\u2005\u00c5 cf. 2.8575\u2005(2) and 2.8594\u2005(3)\u2005\u00c5]. In the crystal, phenyl-C\u2014H\u22efO(carbon\u00adyl) and carbonyl-O\u22efO(carbon\u00adyl) [2.817\u2005(2)\u2005\u00c5] inter\u00adactions combine to form a supra\u00admolecular chain propagating along [111]; the chains pack without directional inter\u00adactions between them. The carbonyl-O\u22efO(carbon\u00adyl) and other weak contacts have an influence upon the Hirshfeld surfaces with O\u22efH contacts making the greatest contribution, i.e. 37.4% cf. 15.8% for O\u22efO and 15.6% for H\u22efH contacts.The title cluster compound, [Ru R3) have played a major role in the formation and subsequent chemistry of metal carbonyl clusters, often relating to the promising catalytic activity of the products 12 with PR3 leads to Ru3(CO)n12\u00a0\u2013\u00a0(PR3)n, n = 1\u20134, cluster compounds 12, thereby making the cluser more reactive 10[PPh2(C6H4SMe)]8 cluster, only the P atom of the PPh2C6H4SMe ligand is coordinated to the metal centre while the thio\u00admethyl group remains uncoordinated ligand are surprisingly few in number 11PPh2(C6H4SMe-4) (I)Tertiary phosphanes (P3(CO)11PPh2(C6H4SMe-4), (I)3 triangle with one Ru centre being bound, equatorially, by the phosphane ligand. The Ru\u2014Ru bond lengths in the Ru3 triangle are not equivalent with the Ru1\u2014Ru2 bond of 2.8933\u2005(2)\u2005\u00c5 being longer than the Ru1\u2014Ru3 and Ru2\u2014Ru3 bonds of 2.8575\u2005(2) and 2.8594\u2005(3)\u2005\u00c5, respectively. This disparity probably reflects the steric hindrance exerted by the phosphane ligand which occupies the region in the vicinity of the Ru1\u2014Ru2 bond. Some general trends in the geometric parameters involving the carbonyl ligands may be discerned, the relatively high errors in some of the parameters notwithstanding. Thus, the Ru\u2014C bond distances involving carbonyl groups lying in the plane of the Ru3 ring are generally shorter than those occupying positions perpendicular to the plane, with the respective ranges in Ru\u2014C bond lengths being 1.897\u2005(3)\u20131.930\u2005(3)\u2005\u00c5 and 1.937\u2005(2)\u20131.953\u2005(3)\u2005\u00c5. While the Ru\u2014C\u2261O angles are all close to linear, two distinctive ranges in angles are evident. The Ru\u2014C\u2261O angles involving carbonyl groups lying in the plane of the Ru3 ring lie in the range 177.3\u2005(2)\u2013178.7\u2005(2)\u00b0 while the range for the perpendicularly orientated carbonyl groups is 172.1\u2005(2)\u2013174.6\u2005(2)\u00b0. The trend for longer Ru\u2014C distances and greater deviations from linearity of the Ru\u2014C\u2261O angles for the axial carbonyl ligands, which occupy positions trans to other carbonyl ligands, is consistent with some semi-bridging character for these carbonyl ligands. Thus, the closest intra\u00admolecular Ru\u22efC(carbon\u00adyl) contact of 3.233\u2005(3)\u2005\u00c5 is formed by the C8-carbonyl ligand which exhibits the maximum deviation from linearity, i.e. 172.1\u2005(2)\u00b0.The mol\u00adecular structure of Rua. The O3\u22efO3i separation is 2.817\u2005(2)\u2005\u00c5, a distance less than the sum of the van der Waals radii of oxygen, i.e. 3.04\u2005\u00c5 : 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z. Such inter\u00admolecular O\u22efO inter\u00adactions are examples of homoatomic chalcogen bonding which are rarest for the smaller oxygen atoms et al., 2017dnorm are shown in Fig.\u00a03a, results from the C21\u2014H\u22efO8 inter\u00adaction contact is evident. Carbon\u00adyl\u22ef\u03c0(arene) inter\u00adactions are known to be important in the structural chemistry of metal carbonyls separation is 3.850\u2005(3)\u2005\u00c5 and the angle subtended at the O4 atom is 90.1\u2005(2)\u00b0, indicating a side-on approach between the residues. The environment about a reference mol\u00adecule, showing short inter\u00adatomic O\u22efO and C\u22efH/H\u22efC contacts significant in the mol\u00adecule packing of (I)The Hirshfeld surface calculations of (I)n Table\u00a01. The preet al., 2007i.e. 15.6%, contribution from these contacts to the Hirshfeld surfaces is due to the presence of the carbonyl groups on the Ru-cluster which leads to an increase in the contribution of O\u22efH/H\u22efO contacts to the Hirshfeld surface, i.e. 37.4%. The single tip at de + di \u223c2.4\u2005\u00c5 in the H\u22efH delineated fingerprint plot, which has a broad appearance, arises from a van der Waals contact between the methyl-H18B and phenyl-H20 atoms s Table\u00a02. The twoe Table\u00a01 and a she Table\u00a01, respecte Table\u00a01 and othee Table\u00a01 are views Table\u00a02 result iChemical context, there are two other Ru3 clusters in the literature having the same (4-methyl\u00adsulfanylphen\u00adyl)di\u00adphenyl\u00adphosphane ligand as in (I)3(CO)9PPh2(C6H4SMe-4)(Ph2PCH2PPh2) (II) 9PPh2(C6H4SMe-4)(Ph2AsCH2AsPh2) 11PRR\u2032R\u2032\u2032 and several examples where the phosphane ligand is bidentate bridging, i.e. Ru3(CO)11PR(R\u2032)\u2013R\u2032\u2032\u2013(R\u2032)RPRu3(CO)11 11PRR\u2032R\u2032\u2032.As mentioned in the 3(CO)12 was purchased from Aldrich and PPh2C6H4SMe was synthesized as reported previously 11P(C6H4SMe-4)Ph2 (I)3(CO)12 and PPh2(C6H4SMe) in tetra\u00adhydro\u00adfuran (25\u2005ml). The reaction mixture was treated dropwise with sodium di\u00adphenyl\u00adketyl solution until the colour of the mixture turned from orange to dark red and then stirred for 30\u2005min. The solvent was evaporated under vacuum and the residue was chromatographed by preparative TLC. Elution with 7:3 n-hexa\u00adne/di\u00adchloro\u00admethane mixture gave four bands and the major orange fraction was characterized as (I)30H17O11PRu3S: C, 39.18; H, 1.86%. Found: C, 39.60; H, 1.90%. IR (C6H12): \u03bd(CO) 2097(m), 2059(w), 2046(m), 2015(s), 1989(w) cm\u22121. 1H NMR (CDCl3): \u03b4 7.45\u20137.23 , 2.48 . 13C NMR (CDCl3): \u03b4 204.24 (Ru\u2014CO), 135.19\u2013125.37 (Ph), 14.79 (Me). 31P NMR (CDCl3): \u03b4 34.28 (s).All reactions were carried out under an inert atmosphere of oxygen-free nitro\u00adgen (OFN) using standard Schlenk techniques. RuUiso(H) set to 1.2\u20131.5Ueq(C). Owing to poor agreement, four reflections, i.e. (1 7 14), I, global. DOI: 10.1107/S2056989018006989/hb7749Isup2.hklStructure factors: contains datablock(s) I. DOI: 1842044CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title compound is constructed from two aromatic rings , which are linked by a C=C\u2014C(=O)\u2014C enone bridge and form a dihedral angle of 17.91\u2005(17)\u00b0. In the crystal, mol\u00adecules are linked by C\u2014H\u22efO hydrogen bonds enclosing rings of 17H14BrFO3, the aromatic rings are tilted with respect to the enone bridge by 13.63\u2005(14) and 4.27\u2005(15)\u00b0, and form a dihedral angle 17.91\u2005(17)\u00b0. In the crystal, centrosymmetrically related mol\u00adecules are linked by pairs of C\u2014H\u22efO hydrogen bonds into dimeric units, forming rings of R22(14) graph-set motif. The dimers are further connected by weak C\u2014H\u22efO hydrogen inter\u00adactions, forming layers parallel to (10In the mol\u00adecule of the title compound, C Chalcones, which are considered to be the precursors of flavonoids and isoflavonoids, are abundant in edible plants. They consist of open-chain flavonoids in which the two aromatic rings are joined by a three-carbon \u03b1,\u03b2-unsaturated carbonyl system. These are coloured compounds because of the presence of the \u2013CO\u2014CH=CH\u2013 chromophore, the colour depending on the presence of other auxochromes. Accumulating evidence has shown that chalcones and their derivatives could inhibit tumor initiation and progression. In view of the above and in a continuation of our previous work on 3,4-dimeth\u00adoxy chalcones \u2005\u00c5 for C16 and \u22120.124\u2005(4)\u2005\u00c5 for C17. The bond lengths and angles are comparable with those found in the related compounds (2E)-3-(3-chloro\u00adphen\u00adyl)-1--prop-2-en-1-one -3--1-prop-2-en-1-one -1-(3-bromo\u00adphen\u00adyl)-3-prop-2-en-1-one -3-(2-bromo\u00adphen\u00adyl)-1-prop-2-en-1-one \u2005\u00c5, C14\u22efCg1i = 4.010\u2005(4)\u2005\u00c5, C14\u2014Br1\u22efCg1i = 82.54\u2005(11)\u00b0; symmetry code: (i) \u22121\u00a0+\u00a0x, y, z; Cg1 is the centroid of the C1\u2013C6 ring] and C\u2014F\u22ef\u03c0 inter\u00adactions help to stabilize the crystal structure.In addition, weak C\u2014Br\u22ef\u03c0 [C14\u2014Br1 = 1.877\u2005(3)\u2005\u00c5, Br1\u22efCrystalExplorer17.5 . The O\u22efH/H\u22efO, Br\u22efC/C\u22efBr and F\u22efC/C\u22efF contacts in the structure with 17.9, 5.6 and 5.0% contributions, respectively, to the Hirshfeld surface have a symmetrical distribution of points . The other Br\u22efC / C\u22efBr, F\u22efC / C\u22efF, C\u22efC, F\u22efO / O\u22efF and C\u22efO / O\u22efC contacts, having only small contributions to the Hirshfeld surface, have negligible directional impact on the mol\u00adecular packing.Mol\u00adecular Hirshfeld surfaces = 1.2Ueq(C) or 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018009416/rz5240sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018009416/rz5240Isup2.hklStructure factors: contains datablock(s) I. DOI: 1852842CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, mol\u00adecules are linked via pairs of the weak inter\u00admolecular C\u2014H\u22efN hydrogen bonds, forming inversion dimers with b]pyridine ring systems.The imidazopyridine unit linked to phenyl and allyl substituents. The allyl substituent is rotated significantly out of the imidazopyridine plane, while the benzene ring is inclined by 3.84\u2005(6)\u00b0 to the ring system. In the crystal, mol\u00adecules are linked via a pair of weak inter\u00admolecular C\u2014H\u22efN hydrogen bonds, forming an inversion dimer with an R22(20) ring motif. The dimers are further connected by \u03c0\u2013\u03c0 stacking inter\u00adactions between the imidazopyridine ring systems [centroid\u2013centroid distances = 3.7161\u2005(13) and 3.8478\u2005(13)\u2005\u00c5]. The important contributions to the Hirshfeld surface are H\u22efH (35.9%), H\u22efCl/Cl\u22efH (15.0%), H\u22efC/C\u22efH (12.4%), H\u22efBr/Br\u22efH (10.8%), H\u22efN/N\u22efH (7.5%), C\u22efBr/Br\u22efC (5.9%), C\u22efC (5.5%) and C\u22efN/N\u22efC (4.0%) contacts.The title compound, C Their activities include anti\u00adcancer -4H-imidazopyridine in the presence of a catalytic qu\u00adantity of tetra-n-butyl\u00adammonium bromide under mild conditions.As a continuation of our research work devoted to the development of substituted imidazopyridine unit linked to phenyl and allyl substituents \u2005\u00c5 for atom C12. The ring system is inclined by 3.84\u2005(6)\u00b0 to the benzene C1\u2013C6 ring, with the N2\u2014C7\u2014C6\u2014C1 torsion angle being 3.3\u2005(3)\u00b0. The allyl substituent is nearly perpendicular to the imidazopyridine plane, as indicated by the C8\u2014N3\u2014C13\u2014C14 torsion angle of \u221297.3\u2005(2)\u00b0. Atoms C6 and C13 are 0.038\u2005(2) and 0.014\u2005(2)\u2005\u00c5, respectively, away from the imidazopyridine plane.The title compound is built up from an imidazopyridine ring systems. The centroid\u2013cen\u00adtroid distances, Cg1\u22efCg1ii and Cg1\u22efCg2i , are 3.7161\u2005(13) and 3.8478\u2005(13)\u2005\u00c5, respectively, where Cg1 and Cg2 are the centroids of the N1/N2/C7\u2013C9 and N3/C8\u2013C12 rings, respectively.In the crystal, mol\u00adecules are linked if Fig.\u00a02. The dimpara-substituated analogue, namely 4-allyl-6-bromo-2-phenyl-4H-imidazopyridine monohydrate, has been reported \u2013(l), together with their relative contributions to the Hirshfeld surface. The contributions are 35.9, 15.0, 12.4, 10.8, 7.5, 5.9, 5.5, 4.0, 1.5, 1.2 and 0.2%, respectively, for H\u22efH, H\u22efCl/Cl\u22efH, H\u22efC/C\u22efH, H\u22efBr/Br\u22efH, H\u22efN/N\u22efH, C\u22efBr/Br\u22efC, C\u22efC, C\u22efN/N\u22efC, C\u22efCl/Cl\u22efC, N\u22efBr/Br\u22efN and N\u22efN contacts. The most important inter\u00adaction is H\u22efH (35.9%), which is reflected as widely scattered points of high density due to the large hydrogen content of the mol\u00adecule [Fig.\u00a05b)]. The spike with the tip at de = di = 1.16\u2005\u00c5 is due to the short inter\u00adatomic H\u22efH contacts. The H\u22efCl/Cl\u22efH contacts (15.0%) have a nearly symmetrical distribution of points and a pair of spikes with tips at de + di = 2.67\u2005\u00c5 [Fig.\u00a05c)]. In the absence of C\u2014H \u22ef \u03c0 inter\u00adactions, the H\u22efC/C\u22efH contacts (12.4%) also have a nearly symmetrical distribution of points with tips at de + di = 2.79\u2005\u00c5 [Fig.\u00a05d)]. The H\u22efBr/Br\u22efH contacts (10.8%) have a symmetrical distribution of points and a pair of spikes with tips at de + di = 3.00\u2005\u00c5 [Fig.\u00a05e)]. A pair of spikes with tips at de + di\u00a0= 2.42\u2005\u00c5 in the H\u22efN/N\u22efH contacts (7.5%) arises from the C\u2014H\u22efN hydrogen bond ]. The C\u22efC contacts (5.5%) have an arrow-shaped distribution of points with the tip at de = di = 1.75\u2005\u00c5 [Fig.\u00a05h)]. The C\u22efN/N\u22efC contacts (4.0%) have wide spikes with tips at de + di = 3.44\u2005\u00c5 [Fig.\u00a05i)]. The HS representations with the function dnorm plotted onto the surface are shown for the H\u22efH, H\u22efCl/Cl\u22efH, H\u22efC/C\u22efH, H\u22efBr/Br\u22efH, H\u22efN/N\u22efH, C\u22efBr/Br\u22efC, C\u22efC and C\u22efN/N\u22efC contacts [Figs.\u00a06a)\u2013(h)].In order to visualize the inter\u00admolecular inter\u00adactions in the crystal of the title compound, a Hirshfeld surface (HS) analysis dissolved in 25\u2005ml of N,N-di\u00admethyl\u00adformamide (DMF) and potassium carbonate was stirred for 5\u2005min, and then to a mixture of tetra-n-butyl\u00adammonium bromide and allyl bromide was added. Stirring was continued for 6\u2005h at room temperature. After removing the salts by filtration, DMF was evaporated under reduced pressure, and the solid obtained was dissolved in di\u00adchloro\u00admethane. The residue was extracted with distilled water and the resulting mixture was chromatographed on a silica-gel column . Brown single crystals suitable for X-ray diffraction were obtained by evaporation of an ethyl acetate\u2013hexane (1:3 v/v) solution.A mixture of 6-bromo-2-(4-chloro\u00adphen\u00adyl)-4Uiso(H) = 1.2Ueq(C).Crystal data, data collection and refinement details are summarized in Table\u00a0210.1107/S2056989018017322/is5505sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018017322/is5505Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018017322/is5505Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018017322/is5505Isup4.cmlSupporting information file. DOI: 1883384CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Similarly, mol\u00adecule (2) presents the same bridging group, but it connects two nearly planar appendants, each comprising a phenyl ring bonded with a hydroxyl and two aldehyde groups. Compound (2) exhibits a strong visible luminescence when excited with ultraviolet radiation.The mol\u00adecule of , and 5,5\u2032-bis\u00ad, C19H16O6, (2), crystallize with one mol\u00adecule in the asymmetric unit. In mol\u00adecule (1), a >C(CH3)2 group bridges two nearly planar salicyl\u00adaldehyde groups [r.m.s deviations = 0.010\u2005(1) and 0.025\u2005(2)\u2005\u00c5], each comprising a planar phenyl ring bonded with a hydroxyl and an aldehyde group. Similarly, compound (2) has the same bridging group, but it connects two nearly planar appendants [r.m.s deviations = 0.034\u2005(1) and 0.035\u2005(1)\u2005\u00c5], each comprising a phenyl ring bonded with a hydroxyl and two aldehyde groups. Mol\u00adecule (1) exhibits a bridge angle of 109.5\u2005(2)\u00b0 with the salicyl\u00adaldehyde planes subtending a dihedral angle of 88.4\u2005(1)\u00b0. In contrast, mol\u00adecule (2) presents a bridge angle of 108.9\u2005(2)\u00b0 with its appendants subtending a dihedral angle of 79.6\u2005(3)\u00b0. Both mol\u00adecules exhibit two intra\u00admolecular O\u2014H\u22efO hydrogen bonds involving the phenolic H atoms and carboxyl O-atom acceptors. In the crystal of (2), O\u2014H\u22efO hydrogen bonds between one of the hydroxyl H atoms and a carboxyl O atom from a symmetry-related mol\u00adecule form a chain along [102) exhibits a strong visible luminescence when excited with ultraviolet radiation.The title compounds 5,5\u2032-bis\u00ad, C These precursor compounds present a >C(CH3)2 group that bridges two salicyl\u00adaldehyde moieties (1) or two phenyl groups with an hydroxyl and two aldehyde appendants (2). The various functional groups in these mol\u00adecules determine their chemical and physical properties, and the ability to modify them provides the title compounds with a wide versatility and the multifunctionality required for synthesizing safer and better performance materials for future civilian and military applications. For instance, the title compounds may be used for the non-toxic, iso\u00adcyanate-free synthesis of polyurethanes is a new, solid-state photoluminescence material that emits radiation in the spectroscopic range between 490 and 590\u2005nm upon ultraviolet light excitation, with potential use as an organic light emitting diode, laser frequency harmonic generator, or photoelectric converter.As polymers play an undeniable role in our everyday lives, extensive resources and safety evaluations are devoted toward the development and marketing of the most suitable and effective polymer species for a given application , the salicyl\u00adaldehyde fragment containing atom C4 (S1A) is near planar [r.m.s. deviation = 0.010\u2005(1)\u2005\u00c5], with a maximum out-of-plane deviation of 0.020\u2005(2)\u2005\u00c5 for the O1 atom. Similarly, its companion salicyl\u00adaldehyde fragment (S1B) is near planar [r.m.s. deviation = 0.025\u2005(2)], with a maximum out-of-plane deviation of 0.050\u2005(2)\u2005\u00c5 for the O3 atom. The bridge angle C4\u2014C1\u2014C11 measures 109.5\u2005(2)\u00b0 and the S1A and S1B planes subtend a dihedral angle of 88.4\u2005(1)\u00b0. Mol\u00adecule (1) exhibits two intra\u00admolecular hydrogen bonds between the phenolic hydrogen atoms and carboxyl O-atom acceptors presents two near planar appendants, denoted A1 and A2 for the appendants containing C4 and C11, respectively, . Each appendant comprises a hydroxyl and two aldehyde groups. Similar to (1), the salicyl\u00adaldehyde fragments with atoms C4\u2013C9/C11/O1/O2 (S2A) or C12\u2013C17/C18/O6/O5 (S2B) in (2) adopt a near planar geometry [r.m.s. deviation = 0.024\u2005(1)\u2005\u00c5 for S2A and 0.036\u2005(1)\u2005\u00c5 for S2B]. The additional carbonyl groups C10\u2014O3 and C18\u2014O4 on the phenyl rings are twisted slightly out of the S2A and S2B planes, respectively, as evidenced by their respective torsion angles C5\u2014C6\u2014C10\u2014O3 [\u22122.9\u2005(4)\u00b0] and C13\u2014C14\u2014C18\u2014O4 [\u2212179.1\u2005(3)\u00b0]. These additional groups increase the steric hindrance between the appendants and methyl bridge groups in (2), perhaps decreasing both the bridge angle C4\u2014C1\u2014C12 [108.9\u2005(2)\u00b0] and the dihedral angle between the A1 and A2 planes [88.4\u2005(1)\u00b0] relative to (1). Mol\u00adecule (2) presents two intra\u00admolecular hydrogen bonds involving the phenolic hydrogen atoms with the carboxyl O-atom acceptors (Table\u00a021).Mol\u00adecule and (2) exhibit several intra\u00admolecular H\u22efH contacts that are shorter than the sum of the H-atom van der Waals radii. These contacts occur between the methyl group H atoms and adjacent phenyl group H atoms . Superimposition of the atoms C1/C2/C3/C4/C11 of (1) with the corresponding atoms of (2) \u00b0 and the S1B and S2B planes subtending an angle of 35.1\u2005(1)\u00b0.Both (1) along the a axis. van der Waals contacts between the O atoms and H atoms of adjacent mol\u00adecules dominate the inter\u00admolecular inter\u00adactions. In addition, bifurcated contacts between atom C17 and atoms H3 and O3 of adjacent mol\u00adecules contribute to the crystal packing. As in mol\u00adecule (1), O\u22efH contacts play a key role in the inter\u00admolecular inter\u00adactions of (2). However, unlike (1), these inter\u00adactions result mostly from hydrogen bonding between the phenolic hydrogen atoms and the carboxyl oxygen atoms of adjacent mol\u00adecules Figs. 4 and 5 \u25b8.et al., 2016et al., 2009et al., 2012et al., 2005et al., 20173), a common chemical known also as bis\u00adphenol A, and 5-[(3-formyl-4-hy\u00addroxy\u00adphen\u00adyl)meth\u00adyl]-2-hy\u00addroxy\u00adbenzaldehyde (4) and (2) and further discussion. Mol\u00adecule (3) presents a submolecular structure of the title compounds, as it only lacks the aldehyde groups found in (1) or (2). In contrast, (4) exhibits a pair of salicyl\u00adaldehyde groups as (1) or (2), except that they are linked by a >CH2 bridge, instead of a >C(CH3)2 bridge.A search of the Cambridge Structural Database crystallizes with three independent mol\u00adecules in the asymmetric unit. Each mol\u00adecule presents a pair of planar phenol fragments subtending dihedral angles of 77.81\u2005(3), 86.15\u2005(4) and 84.34\u2005(4)\u00b0, respectively, and respective bridge angles of 109.2\u2005(1), 109.5\u2005(1), and 108.1\u2005(1)\u00b0. In general, both (1) and (2) have similar geometric parameters to (3), although their corresponding phenol groups are less planar than those of (3). This manifestation results most likely because the phenyl groups of the title compounds contain aldehyde groups in addition to the hydroxyl groups. The O atoms of these aldehyde groups participate in hydrogen bonding with the hydroxyl H atoms, thus partially displacing the hydroxyl O atoms away from the phenol planes. A superimposition of the atoms in (1) with the corresponding atoms of one of the three structures of (3) shows that the differences in the atom positions of the two structures are hardly discernible and its counterpart of (3)]. An overlay of structure (1) onto either structure two or three of (3) yields comparable results. A similar analysis of structures (2) and (3) yields a r.m.s. deviation of 1.14\u2005\u00c5 with maximum displacement of 0.605\u2005(2)\u2005\u00c5 for the C6 atom of (2) and its counterpart in (3). Again, we obtain comparable results overlaying either structure two or three of (3) onto (1).Compound exhibits a pair of near planar salicyl\u00adaldehyde fragments [r.m.s. deviation = 0.0153\u2005(2) and 0.0238\u2005(9)\u2005\u00c5] forming a dihedral angle of 85.96\u2005(4)\u00b0, similar to (1). Its bridge angle of 113.6\u2005(1)\u00b0 is much greater than that of (1) or (2), however. A superimposition of the salicyl\u00adaldehyde group atoms of (4) with corresponding atoms of (1) reveals nearly identical atomic positions of the two groups [r.m.s. deviation = 0.0160\u2005\u00c5], with the companion salicyl\u00adaldehyde group planes [centroid-to-centroid distance measuring = 4.68\u2005(2)\u2005\u00c5] subtending a dihedral angle of 6.81\u00b0. A similar analysis for structures (2) and (4) yields a r.m.s. deviation = 0.027\u2005\u00c5 with companion salicyl\u00adaldehyde groups planes [centroid-to-centroid distance measuring = 4.21\u2005(1)\u2005\u00c5] forming a dihedral angle of 7.4\u2005(1)\u00b0.Mol\u00adecule and (2), respectively).The title compounds were synthesized following modified literature procedures (\u00d6zdemir 1): A combination of compound (3) , paraformaldehyde , and magnesium(II) chloride were suspended in tetra\u00adhydro\u00adfuran , placed under a stream of N2, and stirred. Then, tri\u00adethyl\u00adamine was added dropwise to the reaction mixture at ambient temperature and stirred under reflux for 16\u2005h. At the conclusion of the reaction, the mixture was cooled to room temperature before the addition of diethyl ether (500\u2005mL). The organic solution was sequentially extracted with aqueous 1\u2005M HCl (3 \u00d7 500\u2005mL) and water (3 \u00d7 500\u2005mL), dried over Na2SO\u00ad4 or MgSO4, filtered, and the volatiles were removed under reduced pressure. The solid residue was purified with a series of hexane washes and then dried under vacuum to afford the desired product (1) as a white solid . Slow diffusion of hexa\u00adnes into a benzene solution saturated with (1) afforded single crystals of (1).Compound (2): A mixture of (3) and hexa\u00admethyl\u00adene\u00adtetra\u00admine was dissolved in tri\u00adfluoro\u00adacetic acid under ambient conditions. The reaction mixture was stirred at 403\u2005K for 2.5\u2005h and subsequently cooled to room temperature before aqueous HCl was added slowly. The reaction mixture was stirred at 383\u2005K for 16\u2005h, cooled to room temperature, and the resulting organic phase extracted with di\u00adchloro\u00admethane . Then, this organic phase was dried over MgSO4, filtered, and the volatiles were removed under reduced pressure. The resulting solid was purified with a series of hexa\u00adnes washes and dried under vacuum to afford the novel product (2) as a neon yellow solid . Slow evaporation of a DCM solution saturated with (2) afforded single crystals suitable for X-ray diffractometry.Compound (1) 1H NMR : \u03b4 1.70 , 6.92 , 7.35 , 7.43 , 9.86 , 10.93 ppm. 13C NMR : \u03b4 30.47, 41.86, 117.85, 120.15, 130.92, 136.20, 141.67, 160.10, 196.74 ppm. (2) 1H NMR : \u03b4 1.75 , 7.81 , 10.19 , 11.53 ppm. 13C NMR : \u03b4 30.31, 30.59, 42.08, 123.05, 135.53, 141.31, 162.20, 191.99 ppm; low-resolution mass spectrometry (atmospheric pressure ionization); Thermo Fisher Scientific (ISQ\u2013EC): m/z [M]+: calculated = 340.33; measured: 340; and luminescence spectrum (Horiba Jobin Yvon Fluoro\u00admax 3 Spectrofluorimeter): 10\u22125 \u2005M/aceto\u00adnitrile; \u03bbexc = 356\u2005nm; \u03bbem = 539\u2005nm .Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker 400\u2005MHz spectrometer. Chemical shifts (\u03b4) are given in ppm: (1) and most in (2) were refined in a riding-model approximation with C\u2014H = 0.93 or 0.96\u2005\u00c5, Uiso(H) = 1.2Ueq(C) or 1.5Ueq(Cmeth\u00adyl) and O\u2014H = 0.82\u2005\u00c5 and Uiso(H) = 1.5Ueq(O). In (2), atoms H10, H11, H18, and H5A were refined independently with isotropic displacement parameters.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018016316/lh5885sup1.cifCrystal structure: contains datablock(s) 1, 2. DOI: 10.1107/S2056989018016316/lh58851sup4.hklStructure factors: contains datablock(s) 1. DOI: 10.1107/S2056989018016316/lh58852sup5.hklStructure factors: contains datablock(s) 2. DOI: 1879532, 1879531CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, two mol\u00adecules are associated into an inversion dimer via a pair of C\u2014H\u22ef\u03c0 inter\u00adactions. The dimers are linked by another pair of C\u2014H\u22ef\u03c0 inter\u00adactions, forming a ribbon along the c-axis direction.In the title compound, C 27H29BrN2, the carbazole ring system is essentially planar, with an r.m.s. deviation of 0.0781\u2005(16)\u2005\u00c5. An intra\u00admolecular N\u2014H\u22efN hydrogen bond forms an S(6) ring motif. One of the tert-butyl substituents shows rotational disorder over two sites with occupancies of 0.592\u2005(3) and 0.408\u2005(3). In the crystal, two mol\u00adecules are associated into an inversion dimer through a pair of C\u2014H\u22ef\u03c0 inter\u00adactions. The dimers are further linked by another pair of C\u2014H\u22ef\u03c0 inter\u00adactions, forming a ribbon along the c-axis direction. A C\u2014H\u22ef\u03c0 inter\u00adaction involving the minor disordered component and the carbazole ring system links the ribbons, generating a network sheet parallel to (100).In the title compound, C Thus, the title compound has two tert-butyl groups on the carbazole moiety. The title compound is a suitable model to investigate an intra\u00admolecular hydrogen bond between the heteroaromatic N\u2014H and the N atom of the imino group. We report herein on its mol\u00adecular and crystal structures.Carbazole derivatives have been widely applied in various fields such as pharmaceuticals \u2005\u00c5 at atom C8. There is an intra\u00admolecular N\u2014H\u22efN hydrogen bond involving the amino group (N3\u2014H3) in the carbazole ring and an imine N atom (N2), generating an S(6) ring motif and 0.408\u2005(3).The mol\u00adecular structure of the title compound is shown in Fig.\u00a01f Table\u00a01. The dihA\u2014H22C\u22efCg1i in the major disorder component or C21B\u2014H21E\u22efCg1i in the minor disorder component; Cg1 is the centroid of the C25\u2013C30 ring; symmetry code as in Table\u00a01Cg2ii; Cg2 is the centroid of the C4\u2013C9 ring; symmetry code as in Table\u00a01c-axis direction bis\u00ad[N-methanimine] and 4-bromo\u00adaniline were treated in xylene (10\u2005ml) at 423\u2005K under inert gas overnight, followed by evaporation. The recrystallization of the residue from a solvent mixture of acetone and methanol afforded single crystals of the title compound suitable for X-ray structure analysis . 1H NMR \u03b4 = 1.47 , 1.49 , 7.22 , 7.47\u20137.58 , 7.67 , 8.13 , 8.26 , 8.72 , 10.55 . HR\u2013MS (m/z): calculated for [C27H30BrN2]+, m/z = 461.1587; found, 461.1627.3,6-Di-Uiso(H) = 1.2Ueq(C). Orientational disorder of the tert-butyl substituent (C20\u2013C23) around the C13\u2014C20 bond axis is observed and the occupancies refined to 0.592\u2005(3) and 0.408\u2005(3).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019012374/is5522sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019012374/is5522Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019012374/is5522Isup3.cmlSupporting information file. DOI: 1951647CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "R)- and (S)-camphor thio\u00adsemicarbazone, which crystallizes in the centrosymmetric space group C2/c, is reported.A racemic mixture of ( R)- and (S)-camphorquinone with thio\u00adsemicarbazide yielded the title compound, C11H17N3OS [common name: (R)- and (S)-camphor thio\u00adsemicarbazone], which maintains the chirality of the methyl\u00adated chiral carbon atoms and crystallizes in the centrosymmetric space group C2/c. There are two mol\u00adecules in general positions in the asymmetric unit, one of them being the (1R)-camphor thio\u00adsemicarbazone isomer and the second the (1S)- isomer. In the crystal, the mol\u00adecular units are linked by C\u2014H\u22efS, N\u2014H\u22efO and N\u2014H\u22efS inter\u00adactions, building a tape-like structure parallel to the (R21(7) and R22(8) graph-set motifs for the H\u22efS inter\u00adactions. The Hirshfeld surface analysis indicates that the major contributions for crystal cohesion are from H\u22efH (55.00%), H\u22efS (22.00%), H\u22efN (8.90%) and H\u22efO (8.40%) inter\u00adactions.The equimolar reaction between a racemic mixture of ( R1R2C=O group and it was pointed out that the R1R2C=N\u2014N(H)C(=S)NH2 compound was the main product of the condensation reaction and soft (S) donor atoms in chain chemistry can be traced back to the beginning of the 20th century, when thio\u00adsemicarbazide was used for the chemical characterization of the et al., 2009II, ZnII and FeII/III and reducing the bioavailability of these essential metals, which impacts the growth of tumor cells inhibition in vitro, a key biological target for cancer research - and the other the (1S)-isomer. For the first mol\u00adecule, the 1R and the 4S chiral centers are labelled C2 and C5, and the thio\u00adsemicarbazone unit is nearly planar with a N1\u2014N2\u2014C11\u2014N3 torsion angle of \u22124.7\u2005(2)\u00b0 \u00b0 \u00b0 Fig.\u00a01. In the )\u00b0 Fig.\u00a02. The twoi, N3\u2014H17\u22efO1ii, C5\u2014H5\u22efS2i and N5\u2014H32\u22efS1iii inter\u00adactions and di (x axis) values are the closest external and inter\u00adnal distances from given points on the Hirshfeld surface contacts -camphor 4-phenyl\u00adthio\u00adsemi\u00adcarbazone -camphor 4-phen\u00adyl\u00adthio\u00adsemicarbazone. For the literature structure, the decrease of the contributions from other possible inter\u00adactions is assumed to be due to the geometric impediment of the phenyl ring. The impact of steric effects on the inter\u00admolecular inter\u00adactions sites can be seen in the graphical representation of the Hirshfeld surface in Fig.\u00a09R)-camphor 4-phenyl-TSC mol\u00adecules crystallize as discrete units, being connect by very weak inter\u00adactions. The most frequent inter\u00admolecular inter\u00adactions for the crystal cohesion of the phenyl-TSC derivative are (in %) H\u22efH = 55.9, H\u22efC/C\u22efH = 16.8, H\u22efS/S\u22efH = 11.0, H\u22efO/O\u22efH = 7.8 and H\u22efN/N\u22efH = 7.0. The replace\u00adment of one H atom by the phenyl group in the terminal amine entity strongly impacts on, for example, the contribution of the inter\u00admolecular H\u22efS/S\u22efH inter\u00adactions, which changed from 22.00% to 11.00%. Finally and remarkably, in the comparison mol\u00adecule, inter\u00admolecular H\u22efC/C\u22efH inter\u00adactions make the next highest contibution to the Hirshfeld surface; this inter\u00adaction is comparatively less relevant for the title compound (4.5%).To the best of pur knowledge and from using database tools such as R- and S-camphor was oxidized with SeO2 to the respective 1,2-diketone = 1.2Ueq and C\u2014H bond distances of 0.98\u2005\u00c5 for tertiary carbon atoms and 0.97\u2005\u00c5 for secondary C atoms. The N\u2014H bond distances are 0.86\u2005\u00c5. Finally, Uiso(H) = 1.5Ueq(C) for the methyl groups, with C\u2014H bond distances of 0.96\u2005\u00c5. A rotating model was used for the latter H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019016980/rz5268sup1.cifCrystal structure: contains datablock(s) I, publication_text. DOI: 10.1107/S2056989019016980/rz5268Isup2.hklStructure factors: contains datablock(s) I. DOI: 1973095CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Several O\u2014H\u22efN and N\u2014H\u22efO hydrogen-bonding inter\u00adactions exist between the [Zn(H 2O)6](NO3)2\u00b72C6H5N5, crystallizes in the space group P3\u2212 anion and one half of a [Zn(H2O)6]2+ cation (2O)6]2+ cation and the N atoms of the tetra\u00adzolide ring, and between the nitrate anions and the N\u2014H groups of the pyridinium ring, respectively, giving rise to a three-dimensional network. The 5-(pyridinium-3-yl)tetra\u00adzol-1-ide mol\u00adecules show parallel-displaced \u03c0\u2013\u03c0 stacking inter\u00adactions; the centroid\u2013centroid distance between adjacent tetra\u00adzolide rings is 3.6298\u2005(6)\u2005\u00c5 and that between the pyridinium and tetra\u00adzolide rings is 3.6120\u2005(5)\u2005\u00c5.Hexa\u00adaqua\u00adzinc(II) dinitrate 5-(pyridinium-3-yl)tetra\u00adzol-1-ide, [Zn(H The hexa\u00adaqua\u00adzinc(II) complex exhibits regular octa\u00adhedral geometry 6]X2 cocrystallized with 5-(pyridinium-3-yl)tetra\u00adzol-1-ide tetra\u00adzol-1-ide zwitterion, one NOy Table\u00a01, and they Table\u00a01. The geo2O)6]2+, through O\u2014H\u22efN hydrogen bonds, exhibiting D\u22efA distances in the range 2.7446\u2005(17)\u20132.8589\u2005(17)\u2005\u00c5. Additionally, the pyridinium ring is involved in N\u2014H\u22efO hydrogen bonding to nitrate atom O4, with an N\u22efO distance of 2.7384\u2005(18)\u2005\u00c5. These inter\u00adactions are shown in the crystal packing diagram plane. These parallel-displaced \u03c0\u2013\u03c0 inter\u00adactions lead to inter\u00adplanar distances of 3.21\u2005(1) and 3.10\u2005(3)\u2005\u00c5, and two centroid\u2013centroid distances com\u00adplex cations and nitrate ions serves to hold the structure together Table\u00a02. The N aam Fig.\u00a03. The strs Table\u00a03. The cens Table\u00a03.baab2O)6]X2\u00b72C6H5N5 structures, as more hexa\u00adaqua\u00adzinc(II) complexes can inter\u00adact with the N atoms of the tetra\u00adzole units. Parallel-displaced \u03c0\u2013\u03c0 stacking inter\u00adactions occur in the title compound and in [Mg(H2O)6]X2\u00b72C6H5N5. In [Mg(H2O)6]Cl2\u00b72C6H5N5, the pyridinium\u2013tetra\u00adzolide zwitterions have alternating orientations in the supra\u00admolecular arrangement, whereas in the title compound, the zwitterions are oriented in the same direction, allowing a possible coupling transition between dipole moments similar to J-aggregates cation with a halide counter-ion [chloride and ZnCl2 (2\u2005mmol) were dissolved in 6\u2005ml of distilled water. This mixture was transferred to a glass bottle and then heated at 378\u2005K for 24\u2005h. The pH was adjusted using a HNO3 (66%) solution immediately after mixing the reactants, and was monitored with a pH meter (pH2700 Oakton) until reaching a pH of 2.0. The reaction mixture was then cooled to 318\u2005K and kept at this temperature for 16\u2005h. The colourless block-shaped crystals obtained were washed with ethanol to give 353\u2005mg (yield 30%) of the title compound.All the reactants and chemicals were purchased from Sigma Aldrich and utilized without further purification. A mixture of 3-cyano\u00adpyridine (4\u2005mmol), NaNUiso(H) = 1.2Ueq(C). Moreover, all H atoms in the hexa\u00adaqua\u00adzinc(II) complex were refined with a distance restraint of O\u2014H = 0.85\u2005\u00c5 and with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S205698901801112X/cq2025sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698901801112X/cq2025Isup3.hklStructure factors: contains datablock(s) I. DOI: 1860162CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The ergogenic effect of \u03b2A has also been demonstrated for 2000-m rowing performance prompting interest in whether \u03b2A may be beneficial for sustained aerobic exercise. This study therefore investigated the effect of two \u03b2A dosing strategies on 30-min rowing and subsequent sprint performance.\u2212\u20091, \u03b2A1); ii) matched total \u03b2A ; or iii) cornflour placebo . Participants completed a laboratory 30-min rowing time-trial, followed by 3x30-seconds (s) maximal sprint efforts at days 0, 14 and 28 (T1-T3). Total distance (m), average power (W), relative average power (W\u00b7kg\u2212\u20091), cardio-respiratory measures and perceived exertion were assessed for each 10-min split. Blood lactate ([La-]b mmol\u00b7L\u2212\u20091) was monitored pre-post time-trial and following maximal sprint efforts. A 3-way repeated measures ANOVA was employed for main analyses, with Bonferonni post-hoc assessment (P\u2009\u2264\u20090.05).Following University Ethics approval, twenty-seven healthy, male rowers were randomised in a double-blind manner to 4\u2009weeks of: i) \u03b2A , including absolute average power and relative average power output . These findings were potentially explained by within-group significance for the same variables for the first 10\u2009min split (P\u2009\u2264\u20090.01), and for distance covered (P\u2009=\u20090.01) in the second 10-min split. However, no condition x time interactions were observed. No significant effects were found for sprint variables (P\u2009>\u20090.05) with comparable values at T3 for mean distance , average power and lactate .Total 30-min time-trial distance significantly increased from T1-T3 within \u03b2A1 only may be warranted based on within-group observations.Whilst daily \u03b2A may confer individual benefits, these results demonstrate limited impact of \u03b2A (irrespective of dosing strategy) on 30-min rowing or subsequent sprint performance. Further investigation of \u03b2A dosage >\u20092.4\u2009g\u00b7d Following a 5-min seated period, 2-min baseline expired air samples were collected via the Douglas bag method and anal\u2212]b was assessed following the final collection period only.All time-trials were conducted on the same Concept 2 air braked rowing ergometer with resistance set at 5 for standardisation. Participants undertook a self-paced, continuous warm-up at 100\u2009W for 5-min, after which the unit display was modified to display time remaining to minimise extraneous influences on pacing strategy . Partici\u2212]b, RPE and performance data (distance rowed (m), average power (W)) were recorded on completion of each sprint. At the mid-point of each sprint, standardised verbal encouragement was given to all participants to promote maximal engagement.Following a standardised 5-min inactive rest period, participants completed three 30-s maximal sprint efforts with 60-s inactive recovery in between. b (mmol\u00b7L\u2212\u20091) , absolute VO2 (L\u00b7min\u2212\u20091) or relative VO2 (ml\u00b7kg\u2212\u20091\u00b7min\u2212\u20091) .Intervention groups were matched for age (yrs), rowing experience (yrs), body-height (m), body-mass (kg) and body-fat (%) at baseline Table . Non sigP\u2009=\u20090.22, \u03b7p2\u2009=\u20090.11). A significant effect was shown for time only , with \u03b2A1 distance increasing from 7397\u2009\u00b1\u2009195\u2009m at T1 to 7580\u2009\u00b1\u2009171\u2009m by T3 only .Data for distance rowed (m) during the 30-min time trial are shown as absolute a) and relative b) values in Fig. P\u00a0=\u20090.05, \u03b7p2\u2009=\u20090.12) with \u03b2A1 from 194.8\u00a0\u00b1\u00a018.3\u2009W (T1) to 204.2\u00a0\u00b1\u00a015.5 (T3) (P\u00a0=\u20090.04). No differences were reported within group for \u03b2A2 (200.3\u00a0\u00b1\u00a09.8\u2009W (T1) to 208.8\u00a0\u00b1\u00a08.0\u2009W (T3); P\u00a0>\u20090.05) or PL (173.0\u00a0\u00b1\u00a013.8\u2009W (T1) to 174.8\u00a0\u00b1\u00a013.7\u2009W (T3); P\u2009>\u20090.05). When adjusted for body-mass, average power output expressed as a) absolute and b) relative change is shown in Fig. \u2212\u20091) between T1 and T3 for \u03b2A1 only . A significant between group main effect was reported , with post-hoc analysis indicating an overall difference between \u03b2A2 and PL only (P\u2009=\u20090.04). However, no group x time interactions were shown for absolute changes in average power to weight ratio . When data was expressed as relative change in average power, no significant between group differences were observed ; despite small improvements of 0.13\u2009\u00b1\u20090.06\u2009W\u00b7kg\u2212\u20091 for \u03b2A1 and 0.11\u2009\u00b1\u20090.05\u2009W\u00b7kg\u2212\u20091 for \u03b2A2, in contrast to negligible changes of 0.01\u2009\u00b1\u20090.06\u2009W\u00b7kg\u2212\u20091 for PL.Similarly, mean power significantly increased for time only for \u03b2A1 only , however RPE was maintained throughout the intervention (average RPE: 7.7\u00a0\u00b1\u00a00.2 (T1), 7.8\u00a0\u00b1\u00a00.2 (T2), 7.8\u00a0\u00b1\u00a00.3 (T3) P\u2009>\u20090.05). In contrast, average RPE significantly increased by T3 for both \u03b2A2 (7.1\u00a0\u00b1\u00a00.5 (T1) to 7.7\u00a0\u00b1\u00a00.4 (T3); P\u2009=\u20090.04) and PL (6.5\u00a0\u00b1\u00a00.4 (T1) to 7.3\u00a0\u00b1\u00a00.03 (T3); P\u2009=\u20090.01). Average VO2 was maintained across all trials with no group x time interactions reported (\u03b2A1: 3.12\u00a0\u00b1\u00a00.12\u2009L\u00b7min\u2212\u20091 (T1) to 3.14\u00a0\u00b1\u00a00.10 (T3); \u03b2A2: 3.17\u00a0\u00b1\u00a00.10\u2009L\u00b7min\u2212\u20091 (T1) to 3.24\u00a0\u00b1\u00a00.14 (T3); PL: 2.93\u00a0\u00b1\u00a00.13\u2009L\u00b7min\u2212\u20091 (T1) to 2.86\u00a0\u00b1\u00a00.14 (T3); P\u2009>\u20090.05).Average HR significantly increased by T3 , with \u03b2A1 significantly increasing between T1-T2 (P\u2009=\u20090.03), T1-T3 (P\u2009=\u20090.004) representing a 3.65% (T1-T2) and 4.52% (T1-T3) distance increase, respectively. Likewise, \u03b2A2 also increased distance covered significantly by 2.55% from T2-T3 (P\u2009=\u20090.01). Accompanying these effects for distance, time effects for absolute power was observed in \u03b2A1 only between T1-T2 (P\u2009=\u20090.03) and T1-T3 (P\u2009=\u20090.01). These represented a 14.04 and 14.61% increase in watts, respectively. Time effects for power to weight was observed in \u03b2A1 only, between T1-T2 (P\u2009=\u20090.02) and T1-T3 (P\u2009=\u20090.01). These represented a 6.20 and 6.75% increase in W\u00b7kg\u2212\u20091, respectively.No group x time interactions were shown for overall 0-10\u2009min time trial performance between T1 and T3 for \u03b2A1 and \u03b2A2 , but not PL (P\u2009>\u20090.05). In contrast, average RPE , VE , VO2 , VCO2 and RER were maintained across all trials with no significant interactions reported.A significant effect was observed for changes in HR , with \u03b2A1 increasing distance rowed by 54\u2009\u00b1\u200914\u2009m between T1-T3 (P\u2009=\u20090.01). Other performance variables such as absolute power and power to weight failed to reach significance overall. Average HR , VE , VO2 , VCO2 , RPE , and RER were maintained across all trials with no significant interactions reported.No group x time interactions were shown during the second 10-min split in distance covered or other performance variables (P\u2009>\u20090.05). Average HR , VE , VO2 , VCO2 RPE and RER were maintained across trials with no significant group x time interactions reported.No group x time interactions were shown for overall 21\u201330\u2009min time trial distance covered , or time effects were observed at any time-point for distance covered. Likewise, power , power to weight , HR , RPE and [La-]b failed to reach significance.No significant between-group effects existed at T1 with all groups rowing 166.0\u2009\u00b1\u20092.5\u2009m , on 30-min rowing time trial and subsequent anaerobic sprint performance. A recent 2018 ISSN Position Stand stated tThe findings from the current study support previous research on 10-km running performance , wherebymilieu interieur\u2019 from homeostatic perturbations caused by supra-maximal levels of intracellular acidosis. However, whilst the current cohorts [La\u2212]b were clearly elevated following each sprint bout, the mean [La\u2212]b post time trial and during associated sprint efforts for \u03b2A1 were not significantly affected by \u03b2A consumption compared to PL, possibly suggesting an absence of meaningful carnosine facilitated buffering at the current dosage.Regarding the anaerobic sprint data, no significant interaction effects were observed for any variables. This was unexpected as previously Suzuki et al. noted a \u2212\u20091), it is feasible that one explanation for a lack of significant findings with \u03b2A2 may have been influenced as a result of variances in lean muscle mass.A novel aspect of this study was the inclusion of an alternate day dosing strategy . Previous research has suggested that the primary facilitator of muscle carnosine concentration is the total dose consumed, not factors such as baseline content or daily dose . The curOther reasons for variation caused by dosing may reside in the pharmacokinetics of \u03b2A, with previous research exhibiti\u2212]b, RPE and HR were recorded throughout the test, blood pH was not. Subsequently, it must be conceded that the capacity to assess or infer whether effects were associated with carnosine directed (pH) buffering are limited. Additionally, due to a commonly reported side effect of \u03b2A , particesthesia , 48.\u2212\u20091 for four-weeks is not sufficient to maximise muscle carnosine content [\u2212\u20091 of \u03b2A each day for 24\u2009weeks, observing gene expression, muscle carnosine content and cycling capacity (CCT110%) [\u2212\u20091 [Beyond the delivery method of \u03b2A, previous research demonstrated that at an average daily dose of 5.2\u2009g\u00b7day content . More reCCT110%) . Interes0%) [\u2212\u20091 ) or dura0%) [\u2212\u20091 may be w\u2212\u20091) for longer periods (>\u200928\u2009days) should also be considered.In conclusion, regardless of dose strategy, when compared to placebo, \u03b2A does not enhance sustained aerobic performance or subsequent high intensity efforts. However, the within-group finding that daily \u03b2A use increased 30-min rowing time trial performance warrants further investigation. The inclusion of higher dosing strategies (>\u20092.4\u2009g\u00b7day"} +{"text": "In the crystal, the mol\u00adecules are joined by C\u2014H\u22efO contacts into infinite chains along the 18H23FO5, was synthesized by reacting diethyl malonate with 1-(4-fluoro\u00adphen\u00adyl)-3-methyl\u00adbut-2-en-1-one. The mol\u00adecule adopts a loose conformation stabilized by weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions. In the crystal, the mol\u00adecules are joined by C\u2014H\u22efO contacts into infinite chains along the b-axis direction with a C(6) graph-set motif. Hirshfeld surface analysis and fingerprint plots demonstrate the predominance of H\u22efH, O\u22efH and F\u22efH inter\u00admolecular inter\u00adactions in the crystal structure.The title compound, C The mol\u00adecular conformation is stabilized by an intra\u00admolecular C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 inter\u00adaction \u00b0; the dihedral angles formed by aromatic ring with adjacent and opposite ester groups are 56.66\u2005(4) and 16.08\u2005(4)\u00b0, respectively. The dihedral angle between aromatic ring and ketone carbonyl unit is 14.04\u2005(5)\u00b0.The title compound crystallizes in the monoclinic crystal system in the space group on Fig.\u00a01 and shorB\u22efO2, which join the mol\u00adecules into infinite chains with graph-set motif C(6) \u2014CH2 and C6H4\u2014C(=O)\u2014CH2\u2014CH2\u2014CH(COO)2 gave 102 and 62 hits, respectively. Among them, two hits, (S)-ethyl-2-(4-t-butyl\u00adbenzyl\u00adsulfan\u00adyl)-4-(4-fluoro\u00adphen\u00adyl)-4-oxo\u00adbutano\u00adate -2-(1-(4-nitro\u00adphen\u00adyl)-1,4-dioxo\u00adpentan-3-yl) malonate . Single crystals of the title compound were obtained by slow evaporation from acetone solvent at room temperature.To a stirred solution of diethyl malonate in tetra\u00adhydro\u00adfuran (5\u2005ml), sodium hydride was added at 273\u2005K. The reaction mixture was allowed to stir for 15\u2005min. A solution of 1-(4-fluoro\u00adphen\u00adyl)-3-methyl\u00adbut-2-en-1-one in THF was added into the reaction mixture. The reaction mixture was then allowed to stir overnight at room temperature. The completion of the reaction was monitored by thin layer chromatography. The reaction mixture was quenched with saturated ammonium chloride and extracted with ethyl acetate (2 \u00d7 25\u2005ml). The combined organic layer was washed with water (2 \u00d7 25\u2005ml), brine (25\u2005ml), dried over sodium sulfate and evaporated under reduced pressure to obtain the crude product, which was purified by column chromatography using 60\u2013120 mesh silica gel with ethyl acetate and hexane eluent (Uiso = 1.2 or 1.5Ueq(C). The methyl groups were allowed to rotate.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018012094/yk2116sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018012094/yk2116Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018012094/yk2116Isup3.cmlSupporting information file. DOI: 1863914CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the title pyridiniminium halide salt comprise of one cation and one anion. In the crystal, mol\u00adecules are linked by N\u2014H\u22efBr and C\u2014H\u22efO hydrogen bonds, C\u2014H\u22ef\u03c0 inter\u00adactions, and \u03c0\u2013\u03c0 inter\u00adactions into layers. The inter\u00admolecular inter\u00adactions in the crystal structure are qu\u00adanti\u00adfied by Hirshfeld surface analysis. 17H23N2O+\u00b7Br\u2212, the adamantyl moiety and the pyridiniminium ring are inclined to the ketone bridge by torsion angles of \u221278.1\u2005(2) (C\u2014C\u2014C=O) and 58.3\u2005(2)\u00b0 (C\u2014C\u2014N\u2014C), respectively, and the ketone bridge has a C\u2014C\u2014C\u2014N torsion angle of 174.80\u2005(15)\u00b0. In the crystal, the cations are connected into chains parallel to the c axis by C\u2014H\u22efO hydrogen bonds. The chains are further linked into layers parallel to the bc plane by N\u2014H\u22efBr and C\u2014H\u22efBr hydrogen bonds, C\u2014H\u22ef\u03c0 inter\u00adactions and \u03c0\u2013\u03c0 stacking inter\u00adactions [centroid-to-centroid distance = 3.5657\u2005(11)\u2005\u00c5]. A Hirshfeld surface analysis, which comprises the dnorm surface, electrostatic potential map and two-dimensional fingerprint plots, was carried out to verify the contribution of the various inter\u00admolecular inter\u00adactions.In the cation of the title salt, C The ketone bridge is in an anti\u00adperiplanar conformation [C1\u2014C11\u2014C12\u2014N1 = 174.80\u2005(15)\u00b0]. The dihedral angle formed by the pyrimidinium ring with the ketone bridge is 59.77\u2005(14)\u00b0. Bond lengths and angles in the cation are within normal ranges \u2005\u00c5], indicating partial double-bond character. Similar bond lengths are found in related compounds with an N+=C double bond \u2005\u00c5].In the crystal, the cations are linked into chains along the s Table\u00a01. The chaCrystalExplorer3.1 in the electrostatic potential map indicates hydrogen-acceptor potential, whereas the hydrogen donors are represented by positive electrostatic potential (blue region) . This observation is further confirmed by the respective electrostatic potential maps where Br1 shows negative electrostatic potential as a hydrogen acceptor . Beside those two short inter\u00admolecular contacts, the C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions are shown as light-red spots on the dnorm surface . The reciprocal H\u22efBr/Br\u22efH and H\u22efO/O\u22efH inter\u00adactions with 15.9% and 7.6% contributions, respectively are present as sharp symmetrical spikes at de + di \u2243 2.4 and 2.5\u2005\u00c5, respectively . The reciprocal H\u22efC/C\u22efH inter\u00adactions appear as two symmetrical narrow wings at de + di \u2243 2.5\u2005\u00c5 and contribute 7.8% to the Hirshfeld surface . The reciprocal N\u22efH/H\u22efN interactions appear as a symmetrical V-shaped wing in the FP map with de + di \u2243 2.7\u2005\u00c5 and contribute 2.7% to the Hirshfeld surface . The percentage contributions for other inter\u00admolecular contacts are less than 2.6%.A qu\u00adanti\u00adtative analysis of the inter\u00admolecular inter\u00adactions can be made by studying the fingerprint plots (FP); characteristic pseudo-symmetry wings in the FP Fig.\u00a06. The mos\u2005\u00c5 Fig.\u00a06b. The rly Fig.\u00a06c and 6ece Fig.\u00a06d. The rce Fig.\u00a06f. The pet al., 2017et al., 2018A mixture of 1-adamantly bromo\u00admethyl ketone and 4-amino\u00adpyridine was dissolved in 10\u2005ml of toluene at room temperature, followed by stirring at 358\u2005K for 18\u2005h. The completion of the reaction was marked by the amount of the separated solid from the initially clear and homogeneous mixture of the starting materials. The solid was filtered and washed by ethyl acetate. The final pyridiniminium salt was obtained after the solid had been dried under reduced pressure to remove all volatile organic compounds (Said Uiso(H) = 1.2Ueq(C). The N-bound H atoms were located in a difference-Fourier map and freely refined. One outlier (100) was omitted in the last cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018009131/rz5239sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018009131/rz5239Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018009131/rz5239Isup3.cmlSupporting information file. DOI: 1851334CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "A weak C\u2014Hmeth\u00adoxy\u00admeth\u00adyl\u22ef\u03c0 inter\u00adaction is also observed.The title compound is built up by two dioxolo, two pyridine, one pyridazine and one pyran rings. The two dioxolo rings are in envelope conformations, while the pyran ring is in twisted-boat conformation. The pyradizine ring is oriented at dihedral angles of 9.23\u2005(6) and 12.98\u2005(9)\u00b0 with respect to the pyridine rings, while the dihedral angle between the two pyridine rings is 13.45\u2005(10)\u00b0. In the crystal, C\u2014H 27H30N4O6\u00b7H2O, the two dioxolo rings are in envelope conformations, while the pyran ring is in a twisted-boat conformation. The pyradizine ring is oriented at dihedral angles of 9.23\u2005(6) and 12.98\u2005(9)\u00b0 with respect to the pyridine rings, while the dihedral angle between the two pyridine rings is 13.45\u2005(10)\u00b0. In the crystal, O\u2014Hwater\u22efOpyran, O\u2014Hwater\u22efOmeth\u00adoxy\u00admeth\u00adyl and O\u2014Hwater\u22efNpyridazine hydrogen bonds link the mol\u00adecules into chains along [010]. In addition, weak C\u2014Hdioxolo\u22efOdioxolo hydrogen bonds and a weak C\u2014Hmeth\u00adoxy\u00admeth\u00adyl\u22ef\u03c0 inter\u00adaction complete the three-dimensional structure. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (55.7%), H\u22efC/C\u22efH (14.6%), H\u22efO/O\u22efH (14.5%) and H\u22efN/N\u22efH (9.6%) inter\u00adactions. Hydrogen-bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing. Electrochemical measurements are also reported.In the title compound, C O-triphosphate can be used as a potential substrate for fluorescence detection and imaging of DNA pyridazine and its derivatives has been a goal of chemists in recent years. 5--2\u2032-de\u00adoxy\u00aduridine-5\u2032-B (O2/O3/C2\u2013C4) and C (O4/O5/C5\u2013C7), are in envelope conformations. Atoms O3 and O4 are at the flap positions and are displaced by 0.442\u2005(2) and \u22120.397\u2005(2)\u2005\u00c5, respectively, from the least-squares planes of the four atoms. A puckering analysis of the pyran ring A (O1/C1/C2/C4\u2013C6), gave the parameters QT = 0.6508\u2005(25)\u2005\u00c5, q2 = 0.6451\u2005(25)\u2005\u00c5, q3 = \u22120.0865\u2005(26)\u2005\u00c5, \u03c6 = 214.6\u2005(2)\u00b0 and \u03b8 = 97.64\u2005(23)\u00b0, indicating a twisted-boat conformation. The pyradizine ring D (N1/N2/C14\u2013C17) is oriented at dihedral angles of 9.23\u2005(6) and 12.98\u2005(9)\u00b0, respectively, to the pyridine rings E (N3/C18\u2013C22) and F (N4/C23\u2013C27), while the dihedral angle between the two pyridine rings is 13.45\u2005(10)\u00b0. The meth\u00adoxy\u00admethyl moiety is nearly co-planar with the pyradizine ring, as indicated by the O6\u2014C13\u2014C14\u2014C15 torsion angle of \u2212172.8\u2005(2)\u00b0.The title mol\u00adecule contains two dioxolo, two pyridine, one pyridazine and one pyran rings Fig.\u00a01. The pyrwater\u22efOpyran, O\u2014Hwater\u22efOmeth\u00adoxy\u00admeth\u00adyl and O\u2014Hwater\u22efNpyridazine hydrogen bonds analysis have a symmetrical distribution of points, Fig.\u00a05c, with the thin and thick edges at de + di = 2.85 and 2.78\u2005\u00c5. The pair of characteristic wings in the fingerprint plot delineated into H\u22efO/O\u22efH contacts arises from the O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds arises from the O\u2014H\u22efN hydrogen bonds have a wide spike with the tip at de = di = 1.75\u2005\u00c5.The most important inter\u00adaction is H\u22efH, contributing 55.7% to the overall crystal packing, which is reflected in Fig.\u00a05s Table\u00a01 as well s Table\u00a01 and has ts Fig.\u00a05e, 9.6% s Table\u00a01 as well ts Fig.\u00a05g, 2.4% dnorm plotted onto the surface are shown for the H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH and H\u22efN/N\u22efH inter\u00adactions in Fig.\u00a06a\u2013d, respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH and H\u22efN/N\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing = [1 \u2212 Rt(HCl)/Rt(inh)] \u00d7 100, where Rt(inh) and Rt(HCl) are the charge-transfer resistances for MS immersed in HCl, with the title compound and without inhibitor. Nyquist representations of mild steel in 1 M HCl in the absence and presence of the inhibitor system are shown in Fig.\u00a07The effect of the title compound as an inhibitor of the corrosion of mild steel (MS) were studied using electrochemical impedance spectroscopy in the concentration range of 10M HCl without and with various concentrations of the inhibitor. The impedance diagrams obtained have an almost semicircular appearance. This indicates that the corrosion of mild steel in aqueous solution is mainly controlled by a charge-transfer process. The imp\u00adedance parameters are given in Fig.\u00a08Rct is increased to a maximum value of 185 \u03a9 cm2 for the inhibitor, showing a maximum inhibition efficiency of 91% at 10\u22123M. The decrease in Cdl from the HCl acid value of 200\u2005\u00b5F cm\u22122, may be due to the increase in the thickness of the electrical double layer or to a decrease in the local dielectric constant specimens in 1 et al., 2008viz. aqua\u00adbis\u00adcopper(II) bis\u00ad(tri\u00adfluoro\u00admeth\u00adane\u00adsulfonate) pyridazine]bis\u00ad(\u03bc2-azido)\u00addiaza\u00adidodicopper monohydrate] complexes coordinated by 3,6-di(pyridin-2-yl)pyridazine ligands have been reported was added to a solution of 3,6-bis\u00ad(2-pyrid\u00adyl)-1,2,4,5-tetra\u00adzine (4\u2005mmol) in toluene (20\u2005ml). Stirring was continued at room temperature for 4\u2005h. The solvent was removed under reduced pressure. The residue was separated by chromatography on a column of silica gel with ethyl acetate/hexane (1:2) as eluent. Colourless crystals were isolated on evaporation of the solvent (yield: 82%).6-Uiso(H) = 1.5Ueq(C-meth\u00adyl) or 1.2Ueq(C) for all other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989019009848/lh5910sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019009848/lh5910Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019009848/lh5910Isup3.cdxSupporting information file. DOI: 1939591CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II and CoII cations to form a polymeric complex chain propagating along the b-axis direction.The pyridine-2,6-di\u00adcarboxyl\u00adate anions bridge the Ca 7H3NO4)2(H2O)4]\u00b72H2O}n (1), the CoII ion is N,O,O\u2032-chelated by two pyridine-2,6-di\u00adcarboxyl\u00adate anions in a distorted N2O4 octa\u00adhedral geometry, and two carboxyl\u00adate O atoms of pyridine-2,6-di\u00adcarboxyl\u00adate anions bridge tetra\u00adaqua\u00adcalcium(II) units to form polymeric chains propagating along the b-axis direction. In the crystal, O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, and offset \u03c0\u2013\u03c0 stacking inter\u00adactions [inter\u00adcentroid distances = 3.551\u2005(1) and 3.746\u2005(1)\u2005\u00c5] involving inversion-related pyridine rings link the polymeric chains and lattice water mol\u00adecules to form a supra\u00admolecular three-dimensional framework.In the crystal of the title polymeric complex, {[CoCa(C Firstly, the O and N atoms in these ligands made them easy to chelate or bridge metal ions. Secondly, they can be completely or partially deprotonated to generate Hpdc\u2212 or pyc2\u2212, displaying a variety of coordination modes. As a part of our ongoing studies on heterometallic frameworks, we describe here the synthesis and crystal structure of the title complex,1The controllable synthesis of heterometallic polymers, with their fascinating structures and outstanding properties, is still a challenge in crystal engineering \u2005\u00c5, inter\u00adplanar distance = 3.309\u2005(1)\u2005\u00c5, slippage = 1.755\u2005\u00c5; Cg6\u22efCg6viii = 3.551\u2005(1)\u2005\u00c5, inter\u00adplanar distance = 3.279\u2005(1)\u2005\u00c5, slippage = 1.363\u2005\u00c5; Cg5 and Cg6 are the centroids of pyridine rings N1/C1\u2013C5 and N2/C8\u2013C12, respectively; symmetry codes: (vii) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z; (viii) \u2212x\u00a0+\u00a01, \u2212y, \u2212z\u00a0+\u00a01.In the crystal of k Table\u00a02. Within et al., 2016+ and seven Na+, but no alkali earth metal heterometallic coordination polymers. Hence, the title compound 1 is the first reported heterometallic coordination polymer involving the ligand pyridine-2,6-di\u00adcarb\u00adoxy\u00adlic acid, CoII and an alkali earth metal (CaII).A search of the Cambridge Structural Database , Co(CH3COO)2\u00b74H2O and CaCl2 in 15\u2005ml of distilled H2O was stirred for 10\u2005min in air. 0.5 M NaOH was added dropwise and the mixture was turned into a Parr Teflon-lined stainless steel vessel and heated at 423\u2005K for 3\u2005d. Blue [purple in CIF?] block-shaped crystals of 1 were obtained in a yield of 70% .A mixture of HUiso(H) = 1.5Ueq(O). C-bound H atoms atoms were included in calculated positions and refined as riding: C\u2014H = 0.93\u2005\u00c5 with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018007120/xu5921sup1.cifCrystal structure: contains datablock(s) 1, global. DOI: 1832782CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the mol\u00adecules are linked by N\u2014H\u22efO hydrogen bonds into supra\u00admolecular chains propagating along the [101] direction. 16H20N2O6S2, the mid-point of the C\u2014C bond of the central ethane moiety is located on a twofold rotation axis. In the crystal, mol\u00adecules are linked by N\u2014H\u22efO hydrogen bonds into supra\u00admolecular chains propagating along the [101] direction. Hirshfeld surface analysis and two-dimensional fingerprint plots indicate that the most important contributions to the crystal packing are from H\u22efH (43.1%), O\u22efH/H\u22efO (40.9%), C\u22efH/H\u22efC (8.8%) and C\u22efC (5.5%) inter\u00adactions.In the mol\u00adecule of the title compound, C These effective drug mol\u00adecules have an important role in the medical field, including as promising chemotherapeutic agents, and have been used in the treatment of many bacterial infections due to their physical, chemical and biological properties ]bis\u00ad(4-methyl\u00adbenzene\u00adsulf\u00adon\u00ada\u00admide).Sulfonamides are synthetic mol\u00adecules which include the SO2vC (mm2) (H atoms excluded), with the twofold rotation axis bisecting the central C1\u2014C1i bond. The mol\u00adecule is Z-shaped with the N1\u2014S1\u2014C2\u2014C3 torsion angle being \u221260.6\u2005(3)\u00b0. The C1\u2014O1 bond length of 1.429\u2005(3)\u2005\u00c5 and the O1\u2014N1 bond length of 1.426\u2005(2)\u2005\u00c5 are close to the values reported for similar compounds (see the Database survey). The S1\u2014O2 and S1\u2014O3 distances are 1.4376\u2005(17) and 1.4168\u2005(19)\u2005\u00c5, respectively while the S1\u2014N1 and S1\u2014C2 distances are 1.647\u2005(3)\u2005\u00c5 and 1.747\u2005(3)\u2005\u00c5, respectively.The mol\u00adecular structure of the title compound is illustrated in Fig.\u00a013, ca 5% of the unit-cell volume.The crystal packing of the title compound features inter\u00admolecular N\u2014H\u22efO hydrogen bonds Table\u00a01, which cet al., 2016S,2S,4S,5S)-2,5-bis\u00ad[(p-toluene\u00adsulfon\u00adyl)amino]\u00adbicyclo\u00ad(2.2.1)heptane amino]-1-thio\u00adhexitol -1-(toluene\u00adsulfonyl\u00adamido)-3-(toluene\u00adsulfonyl\u00adamido\u00admeth\u00adyl)-3,5,5-tri\u00admethyl\u00adcyclo\u00adhexane surface resolution with the three-dimensional dnorm surfaces mapped over a fixed colour scale of \u22120.464 (red) to 2.052 (blue) \u00c5 using the CrystalExplorer points associated with hydrogen atoms. It is characterized by an end point that points to the origin and corresponds to di = de = 1.08\u2005\u00c5, which indicates the presence of the H\u22efH contacts in this study (43.1%). The C\u22efH/H\u22efC plot in Fig.\u00a07vice versa. There are two symmetrical wings on the left and right sides (8.8%). Furthermore, there are C\u22efC (5.5%), N\u22efH/H\u22efN (1.4%), O\u22efC/C\u22efO (0.1%) and S\u22efH/H\u22efS (0.1%) contacts in the title structure.The H\u22efH plot shown in Fig.\u00a07A view of the three-dimensional Hirshfeld surface of the title compound plotted over electrostatic potential energy in the range \u22120.095 to 0.123 a.u. using the STO-3G basis set at the Hartree\u2013Fock level of theory is shown in Fig.\u00a08The title compound was synthesized according to the method of Bauer & Suresh 1963. Single Uiso(H) = 1.5Ueq(C-methyl) and 1.2Ueq(C) for other H atoms. The NH H atom was located in a difference-Fourier maps and freely refined.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018017437/xu5951sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018017437/xu5951Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018017437/xu5951Isup3.cmlSupporting information file. DOI: 1884045CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the mol\u00adecules are connected by weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22efF hydrogen bonds into sheets parallel to (104). Weak inter\u00admolecular \u03c0\u2013\u03c0 inter\u00adactions also occur.The mol\u00adecular structure of the title chalcone derivative is nearly planar and the mol\u00adecule adopts a 15H10FNO3, is nearly planar and the mol\u00adecule adopts a trans configuration with respect to the C=C double bond. The nitro group is nearly coplanar with the attached benzene ring, which is nearly parallel to the second benzene ring. In the crystal, mol\u00adecules are connected by pairs of weak inter\u00admolecular C\u2014H\u22efO hydrogen bonds into inversion dimers. The dimers are further linked by another C\u2014H\u22efO hydrogen bond and a C\u2014H\u22efF hydrogen bond into sheets parallel to (104). \u03c0\u2013\u03c0 inter\u00adactions occur between the sheets, with a centroid\u2013centroid distance of 3.8860\u2005(11)\u2005\u00c5. Hirshfeld surface analysis was used to investigate and qu\u00adantify the inter\u00admolecular inter\u00adactions.The mol\u00adecular structure of the title chalcone derivative, C This non-linearity leads to frequency-mixing processes , the optical Kerr effect para-substituted phenyl rings and an enone connecting bridge \u00b0. The C7=O3 carbonyl group adopts an s-cis configuration with respect to the C8=C9 double bond as indicated by O3\u2014C7\u2014C8\u2014C9 torsion angle of \u22120.8\u2005(3)\u00b0. The mol\u00adecule (excluding H atoms) is nearly planar with a maximum deviation of 0.103\u2005(2)\u2005\u00c5 at atom O1 of the terminal nitro group. The nitro group is nearly coplanar with the attached C1\u2013C6 benzene ring as indicated by the small dihedral angle of 7.9\u2005(2)\u00b0. The C1\u2013C6 and C10\u2013C15 benzene rings make a small dihedral angle of 4.27\u2005(8)\u00b0 with each other.The asymmetric unit of the title chalcone derivative consists of a unique mol\u00adecule, containing two ge Fig.\u00a01. The molA\u22efO3ii; symmetry code as in Table\u00a01A\u22efO1iii and C4\u2014H4A\u22efF1i; Table\u00a01Cg1\u22efCg1iv,v and Cg2\u22efCg2iv,v = 3.8860\u2005(11)\u2005\u00c5, where Cg1 and Cg2 are the centroids of C1\u2013C6 and C10\u2013C15 benzene rings, respectively; symmetry codes: (iv) x\u00a0\u2212\u00a01, y, z; (v) x\u00a0+\u00a01, y, z] ] correspond to the C11\u2014H11A\u22efO3 hydrogen bond. The C4\u2014H4A\u22efF1 and C15\u2014H15A\u22efO1 hydrogen bonds are indicated as two pairs of lighter red spots on the dnorm surface. The H12A\u22efF1 contact, with its H\u22efF distance shorter than the sum of van der Waals radii by 0.01\u2005\u00c5, appears as two tiny red spots on the dnorm surface. The donor and acceptor of a hydrogen bond with positive and negative electrostatic potentials, respectively, are represented as blue and red regions on the Hirshfeld surface mapped with electrostatic potential [Fig.\u00a04b)]. The electrostatic potential of the F atom is less negative as compared to the O atoms of nitro and carbonyl groups, as indicated by the lighter red region. The H\u22efO/O\u22efH contacts are the most populated contacts and contribute 30.2% of the total inter\u00admolecular contacts, followed by H\u22efH (20.6%), H\u22efC/C\u22efH (18.0%), H\u22efF/F\u22efH (13.1%) and C\u22efC (10.1%) contacts -1-(4-nitro\u00adphen\u00adyl)-3-phenyl\u00adprop-2-en-1-one -3-(4-fluoro\u00adphen\u00adyl)-1-phenyl\u00adprop-2-en-1-one and 4-fluoro\u00adbenzaldehyde were dissolved in methanol (20\u2005ml). A catalytic amount of NaOH was added to the solution dropwise with vigorous stirring. The reaction mixture was stirred for about 6\u2005h at room temperature. The progress of the reaction was monitored by TLC. The formed crude product was filtered, washed repeatedly with distilled water and recrystallized from ethanol to obtain the title chalcone derivative. Yellowish single-crystals suitable for X-ray diffraction were obtained from an acetone solution by slow evaporation at room temperature.Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018017450/is5506sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018017450/is5506Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018017450/is5506Isup3.cmlSupporting information file. DOI: 1036743CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, inversion-related mol\u00adecules form dimers through inter\u00admolecular N\u2014H\u22efO hydrogen bonds with the dimers associated along the 20H17N3O3 , is built up from a benzodiazepine ring system linked to pyridyl and pendant di\u00adhydro\u00adpyran rings, where the benzene and pyridyl rings are oriented at a dihedral angle of 43.36\u2005(6)\u00b0. The pendant di\u00adhydro\u00adpyran ring is rotationally disordered in a 90.899\u2005(3):0.101\u2005(3) ratio with the orientation of each component largely determined by intra\u00admolecular N\u2014HDiazp\u22efODhydp (Diazp = diazepine and Dhydp = di\u00adhydro\u00adpyran) hydrogen bonds. In the crystal, mol\u00adecules are linked via pairs of weak inter\u00admolecular N\u2014HDiazp\u22efODhydp hydrogen bonds, forming inversion-related dimers with R22(26) ring motifs. The dimers are further connected along the b-axis direction by \u03c0\u2013\u03c0 stacking inter\u00adactions between the pendant di\u00adhydro\u00adpyran and pyridyl rings with centroid\u2013centroid distances of 3.833\u2005(3)\u2005\u00c5 and a dihedral angle of 14.51\u2005(2)\u00b0. Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H\u22efH (50.1%), H\u22efC/C\u22efH (17.7%), H\u22efO/O\u22efH (16.8%), C\u22efC (7.7%) and H\u22efN/N\u22efH (5.3%) inter\u00adactions. Hydrogen-bonding and van der Waals inter\u00adactions are the dominant inter\u00adactions in the crystal packing.The title compound, C In fact, many 1,5-benzodiazepines are best known to possess biologically diverse activities such as anti-inflammatory, hypnotic, anti-HIV-1, anti\u00adconvulsant and anti\u00admicrobial (Roma A (C1\u2013C6) is oriented at a dihedral angle of 43.36\u2005(6)\u00b0 with respect to the pyridyl ring C (N3/C10\u2013C14). The pendant di\u00adhydro\u00adpyran ring D (O1/C15\u2013C19) shows a 90.899\u2005(3):0.101\u2005(3) disorder with the minor component rotated by 174.6\u2005(4)\u00b0 from the orientation of the major component. The orientation of both components is largely determined by intra\u00admolecular N2\u2014H2A\u22efO2 or N2\u2014H2A\u22efO3A hydrogen bonds (Table\u00a01D gave the parameters Q = 0.127\u2005(2)\u2005\u00c5, \u03b8 = 108.0\u2005(8)\u00b0 and \u03c6 = 79.6\u2005(8)\u00b0 while for the seven-membered diazepine ring B (N1/N2/C1/C6\u2013C9), the parameters are Q(2) = 0.8888\u2005(13)\u2005\u00c5, Q(3) = 0.2070\u2005(13)\u2005\u00c5, \u03c6(2) = 201.03\u2005(8)\u00b0 and \u03c6(3) = 293.9\u2005(4)\u00b0.The title compound, (I)s Table\u00a01. A puckeDiazp\u22efODhydp (Diazp = diazepine and Dhydp = di\u00adhydro\u00adpyran) hydrogen bonds = 3.833\u2005(3)\u2005\u00c5 with a dihedral angle of 14.51\u2005(2)\u00b0; Cg1 and Cg2 are the centroids of rings D (O1/C15\u2013C19) and C (N3/C10\u2013C14), respectively].In the crystal, the mol\u00adecules are linked via pairs of weak inter\u00admolecular N\u2014Hs Table\u00a01, formingon Fig.\u00a02 by \u03c0\u2013\u03c0-sCrystalExplorer17.5 , and those delineated into H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH, C\u22efC, H\u22efN/N\u22efH, N\u22efC/C\u22efN, O\u22efC/C\u22efO, N\u22efN, N\u22efO/O\u22efN and O\u22efO contacts \u2013(k), respectively, together with their relative contributions to the Hirshfeld surface. H\u22efH inter\u00adactions are the most important, contributing 50.1% to the overall crystal packing, and are shown in Fig.\u00a05b) as widely scattered points of high density because of the large hydrogen content of the mol\u00adecule. The two pairs of thin and thick spikes with the tips at de + di \u223c2.27 and 1.95\u2005\u00c5, respectively, in Fig.\u00a05b) are due to the short inter\u00adatomic H\u22efH contacts (Table\u00a02c), with the tips at de + di \u223c2.82\u2005\u00c5. The two pairs of thin and thick spikes with the tips at de + di = 2.67 and 2.40\u2005\u00c5, respectively, in Fig.\u00a05d) are due to the N\u2014H\u22efO hydrogen bonds (Table\u00a01e)] contacts contribute 7.0% to the HS and have symmetrical distribution of points, with the tips at de + di = 3.24\u2005\u00c5. The pair of characteristic wings in the fingerprint plot delineated into H\u22efN/N\u22efH contacts has a pair of spikes with the tips at de + di = 1.49\u2005\u00c5. Finally, the N\u22efC/C\u22efN contacts [Fig.\u00a05g)] contribute 1.5% to the HS and are viewed as a symmetrical distribution of points with pairs of thin edges at de + di = 3.36\u2005\u00c5.In order to visualize the inter\u00admolecular inter\u00adactions in the crystal of the title compound, a Hirshfeld surface (HS) analysis \u2013(e), respectively.The Hirshfeld surface representations with the function et al., 2015The Hirshfeld surface analysis confirms the importance of H-atom contacts in establishing the packing. The large number of H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH and H\u22efN/N\u22efH inter\u00adactions suggest that van der Waals inter\u00adactions and hydrogen bonding play the major roles in the crystal packing (Hathwar To a suspension of 3-[1-(2-amino\u00adphenyl\u00adimino)\u00adeth\u00adyl]-4-hy\u00addroxy-6-methyl\u00adpyran-2-one (4\u2005mmol) in ethanol (40\u2005ml) were added 1.5 equivalents of 2-pyridine\u00adcarboxaldehyde and three drops of tri\u00adfluoro\u00adacetic acid (TFA). The mixture was refluxed for 4\u2005h. Cooling to room temperature induced the precipitation of a yellow solid, which was filtered off and washed with 20\u2005ml of cold ethanol. Cooling to room temperature induced the precipitation of a yellow solid, which was filtered and washed with 20\u2005ml of cold ethanol. Crystals suitable for X-ray analysis were obrained by recrystallization of the product from ethanol solution.Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018017565/lh5888sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018017565/lh5888Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018017565/lh5888Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018017565/lh5888Isup4.cmlSupporting information file. DOI: 1884597CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "TS3, have been confirmed by resonant scattering. The roles of the water mol\u00adecules and the C\u2014H\u22ef\u03c0 inter\u00adactions in the crystal packing are highlighted.The absolute configurations of the 10 asymmetric carbons involved in the structure of the title limonoid, 26H28O5\u00b70.5H2O (TS3) oxirenocyclo\u00adpenta\u00adphenanthrofuran-3(3aH)-one hemihydrate], crystallizes with two independent mol\u00adecules (1 and 2) in the asymmetric unit and one water mol\u00adecule. TS3 is composed of three six-membered rings , three five-membered rings and two epoxide rings. A group of five fused rings (A\u2013E) is bonded to a furan ring (F) with a Csp3\u2014Csp2 bond [1.500\u2005(3)\u2005\u00c5 in mol\u00adecule 1 and 1.499\u2005(3)\u2005\u00c5 in mol\u00adecule 2]. The absolute structures of the mol\u00adecules in the crystal were determined by resonant scattering; Flack parameter = 0.05\u2005(5). In the crystal, the individual mol\u00adecules stack in columns along the b-axis direction. The water mol\u00adecule bridges mol\u00adecules 1 and 2 via Owater\u2014H\u22efO and C\u2014H\u22efOwater hydrogen bonds. Together with further C\u2014H\u22efO hydrogen bonds, linking mol\u00adecules 1 and 2, the columns are linked to form slabs parallel to the ab plane. Within each column, mol\u00adecules are also linked via C\u2014H\u22ef\u03c0 inter\u00adactions involving the five-membered furan (F) rings.The title limonoid compound, C Rings A to E are fused (first compartment), while ring F is bonded to this first moiety by a Csp3\u2014Csp2 bond, [C15\u2014C19 = 1.500\u2005(3)\u2005\u00c5 and C15B\u2014C19B = 1.499\u2005(3)\u2005\u00c5], as shown in Fig.\u00a01As previously reported, using one- and two-dimensional NMR techniques in combination with high-resolution mass spectroscopy studies /0.672\u2005(2)\u2005\u00c5 for atoms C11/C11B and by 0.654\u2005(2)/0.670\u2005(2)\u2005\u00c5 for atoms C16/C16B. The six-membered ring C has a half-chair conformation in both mol\u00adecules; the puckering parameters for mol\u00adecule 1 are amplitude Q = 0.474\u2005(2)\u2005\u00c5, \u03b8 = 131.7\u2005(2)\u00b0 and \u03c6 = 40.9\u2005(3)\u00b0, while for mol\u00adecule 2 Q = 0.479\u2005(2)\u2005\u00c5, \u03b8 = 127.5\u2005(2)\u00b0, \u03c6 = 42.9\u2005(3)\u00b0. The five-membered rings B and E have envelope conformations with atoms C4/C4B and C15/C15B, respectively, as the flaps, being displaced from the mean plane of the other four atoms by 0.689\u2005(2)/0.702\u2005(2)\u2005\u00c5 and 0.526\u2005(2)/0.454\u2005(2)\u2005\u00c5, respectively. The furan rings (F), are planar in both mol\u00adecules.The six-membered rings TS3 comes from ten asymmetric carbon atoms , refined using Cu K\u03b1 radiation.The chirality of TS3, and details are given in Table\u00a01via three weak hydrogen bonds present in the structure of ds Fig.\u00a03. The indt Table\u00a01, stabilit Table\u00a01 are linket al., 2016TS3 gave no hits. The moieties having the rings E and F have been seen in three cytotoxic limonoids, viz. aphanastatine, amoorastatine and hydroxyl-12-ammorastatine , but having different substituents, are known. Most of these compounds are reported as hemisynthesis products, while TS3 was obtained from a natural source.A search in the Cambridge Structural Database and needle-like crystals, suitable for single crystal X-ray diffraction analysis, were obtained by slow evaporation of the solvents at room temperature after three days.The title compound was isolated from the root bark of al. 2016. A smallUiso(H) = 1.5Ueq(C-meth\u00adyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018009775/su5448sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989018009775/su5448Isup2.hklStructure factors: contains datablock(s) I. DOI: 1854616CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the title mol\u00adecule, di-methyl\u00adhydroxy and 4-nitro\u00adbenzene groups cap a central di-substituted acetyl\u00adene residue. The extended structure features flattened, hexa\u00admeric clusters sustained by hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) hydrogen bonds. 11H11NO3, is capped at either end by di-methyl\u00adhydroxy and 4-nitro\u00adbenzene groups; the nitro substituent is close to co-planar with the ring to which it is attached [dihedral angle = 9.4\u2005(3)\u00b0]. The most prominent feature of the mol\u00adecular packing is the formation, via hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) hydrogen bonds, of hexa\u00admeric clusters about a site of symmetry 6 synthons and have the shape of a flattened chair. The clusters are connected into a three-dimensional architecture by benzene-C\u2014H\u22efO(nitro) inter\u00adactions, involving both nitro-O atoms. The aforementioned inter\u00adactions are readily identified in the calculated Hirshfeld surface. Computational chemistry indicates there is a significant energy, primarily electrostatic in nature, associated with the hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) hydrogen bonds. Dispersion forces are more important in the other identified but, weaker inter\u00admolecular contacts.The di-substituted acetyl\u00adene residue in the title compound, C The aggregates are disposed about a site of symmetry b). The crystal also features weak benzene-C\u2014H\u22efO(nitro) inter\u00adactions, involving both nitro-O atoms. In essence, one nitro group of one mol\u00adecule forms two such inter\u00adactions with two symmetry-related mol\u00adecules to form a supra\u00admolecular chain along the c-axis direction with helical symmetry (31 screw axis), Fig.\u00a03a). An end-on view of the chain is shown in Fig.\u00a03b). These weak benzene-C\u2014H\u22efO(nitro) inter\u00adactions serve to link the six-mol\u00adecule aggregates into a three-dimensional architecture, Fig.\u00a04The spectacular feature of the mol\u00adecular packing of (I)et al., 2019Crystal Explorer 17 , the donors and acceptors of O\u2014H\u22efO hydrogen bond involving the atoms of the hydroxyl group are characterized as bright-red spots. The faint-red spots near the phenyl-H10, H11 and nitro-O2, O3 atoms on the dnorm-mapped Hirshfeld surface in Fig.\u00a05b) represent the effect of weak C\u2014H\u22efO inter\u00adactions as listed in Table\u00a01a), the C\u2014H\u22ef\u03c0/\u03c0\u22efH\u2014C contacts listed in Table\u00a02b) is an indication of the \u03c0\u2013\u03c0 stacking inter\u00adaction between them .The Hirshfeld surface calculations for (I)a), and those delineated into H\u22efH, O\u22efH/H\u22efO, C\u22efH/H\u22efC and C\u22efC contacts \u2013(e), respectively, and provide more information on the influence of short inter\u00adatomic contacts upon the mol\u00adecular packing. The percentage contributions from the different inter\u00adatomic contacts to the Hirshfeld surface are summarized in Table\u00a03de + di \u223c1.8\u2005\u00c5 in the fingerprint plot delineated into O\u22efH/H\u22efO contacts, Fig.\u00a08c), are due to the presence of the O\u2014H\u22efO hydrogen bond, whereas the points corresponding to comparatively weak inter\u00admolecular C\u2014H\u22efO inter\u00adactions, Table\u00a01C atom and the benzene (C6\u2013C11) ring, results in short inter\u00adatomic C\u22efH/H\u22efC contacts, Table\u00a02a), and by the pair of forceps-like tips at de + di \u223c2.8\u2005\u00c5 in Fig.\u00a08d). The points corresponding to other such short inter\u00adatomic contacts involving the acetyl\u00adene-C5 and methyl-C3\u2014H3c atoms at longer separations are merged within the plot. The arrow-shaped distribution of points around de + di \u223c3.6\u2005\u00c5 in the fingerprint plot delineated into C\u22efC contacts, Fig.\u00a08e), indicate \u03c0\u2013\u03c0 overlap between symmetry-related benzene (C6\u2013C11) rings, as illustrated in Fig.\u00a07b). The small percentage contributions from the other inter\u00adatomic contacts listed in Table\u00a03The overall two-dimensional fingerprint plot for (I)Eele), polarization (Epol), dispersion (Edis) and exchange-repulsion (Erep) terms after applying relevant scale factors \u03c1(r), where \u03c1 is the electron density and \u03bb2 is the second eigenvalue of the Hessian matrix of \u03c1. Crucially, through a three-colour scheme, a specific inter\u00adaction can be identified as being attractive or otherwise. Thus, a green isosurface indicates a weakly attractive inter\u00adaction whereas a blue isosurface indicates an attractive inter\u00adaction; a repulsive inter\u00adaction appears red. The isosurfaces for three identified inter\u00admolecular inter\u00adactions are given in the upper view of Fig.\u00a010a), a green isosurface is apparent for the conventional hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) hydrogen bond. Similarly, green isosurfaces are seen between the inter\u00adacting atoms involved in the phenyl-C\u2014H\u22efO(nitro), Fig.\u00a010b), and the methyl-C\u2014H\u22ef\u03c0(C11\u2013C16), Fig.\u00a010c), inter\u00adactions.Non-covalent inter\u00adaction plot (NCIplot) analyses provide a visual representation of the nature of the contact between specified species in crystals (Johnson versus sign(\u03bb2)\u03c1(r). The non-covalent inter\u00adaction peaks appear at density values less than 0.0 atomic units, consistent with their being weakly attractive inter\u00adactions.The lower views of Fig.\u00a010et al., 2010et al., 2016P21/c to form a hexa\u00admeric clusters via hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) hydrogen bonds as seen in (I)a); intra\u00admolecular hy\u00addroxy-O\u2014H\u22efO(carbon\u00adyl) hydrogen bonds are also apparent. In (III), the two independent mol\u00adecules comprising the asymmetric unit associate about a centre of inversion in space group P21/n into a supra\u00admolecular dimer via pairs of hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) and hy\u00addroxy-O\u2014H\u22efN(cyano) hydrogen bonds as shown in Fig.\u00a011b). In this case, one independent hy\u00addroxy-oxygen atom and one cyano-nitro\u00adgen atom do not accept a hydrogen-bonding inter\u00adaction. Three crystallographically independent mol\u00adecules are also found in (II) (space group Pca21) and these self-associate to form a supra\u00admolecular chain via hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) hydrogen bonds with non-crystallographic threefold symmetry, Fig.\u00a011c). Finally, zigzag supra\u00admolecular chains sustained by hy\u00addroxy-O\u2014H\u22efO(hy\u00addroxy) hydrogen bonds are found in the crystal of (IV), Fig.\u00a011d) in space group Pbca.There are four literature precedents for (I)et al., 19981H NMR : \u03b4 = 8.16 , 7.54 , 2.24 and 1.63 ppm. 13C NMR : \u03b4 = 147.2, 132.5, 129.8, 123.6, 99.2, 80.5, 66.7 and 31.3 ppm. Irregular colourless crystals of (I)The title compound was prepared as per the literature procedure (Bleicher Uiso(H) set to 1.2\u20131.5Ueq(C). The O-bound H atom was refined with a distance restraint of 0.82\u00b10.01\u2005\u00c5, and with Uiso(H) = 1.5Ueq(O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989019010284/hb7841sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019010284/hb7841Isup2.hklStructure factors: contains datablock(s) I. DOI: 1941466CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The manganese compound crystallized in the monoclinic space group 1/nP2 and exists as a centrosymmetric dimer.The title compound was prepared by a template method starting from manganese(II) nitrate with a Schiff base ligand. The product of condensation was between methyl carbazate and glyoxylic acid, and formed 2(C4H5N2O4)2(NCS)2(H2O)4]\u00b74H2O (I), exists as a centrosymmetric dimer. Each dimeric unit consists of tridentate -chelating Schiff bases with symmetry-maintained \u03bc-O-bridged carboxyl\u00adate anions, terminally bound thio\u00adcyanate anions, and ligated and solvated water mol\u00adecules. The complex exhibits a distorted octa\u00adhedron geometry and the centrosymmetric \u03bc-O-bridged carboxyl\u00adate anions connect the two manganese atoms to form an M2O2 ring. In the crystal, the mol\u00adecules are inter\u00adlinked via strong N\u2014H\u22efO and O\u2014H\u22efO hydrogen-bonding contacts and weak O\u2014H\u22efS inter\u00admolecular inter\u00adactions, forming a three-dimensional mol\u00adecular network.The title compound, [Mn Among them, carbaza\u00adtes are inter\u00adesting as ligands in view of their variety of potential donor atoms such as oxygen and nitro\u00adgen. Inter\u00adestingly, these neutral mol\u00adecules can be expected to exhibit only one common coordination mode, i.e. N,O-chelating bidentate. This has been clearly observed in many metal complexes with a variety of anions such as formate benzaldehyde with ethyl carbazate and the N-bonded NCS anion, whereas the \u03bc-O-bridged carboxyl\u00adate anions, azomethine nitro\u00adgen atom and a coordinated water mol\u00adecule (O1W) occupy the equatorial positions. The two manganese atoms are connected via centrosymmetrically related \u03bc-O-bridged carboxyl\u00adate anions, forming a rhomboidal Mn2O2 unit about an inversion centre.The manganese title compound crystallizes in the monoclinic space group er Fig.\u00a01. The asyO-bridged carboxyl\u00adate anions are 2.1448\u2005(9), 2.1905\u2005(9) and 2.2606\u2005(8), 2.2985\u2005(8)\u2005\u00c5, respectively. The Mn\u2014N\u2014C\u2014S torsion angle in the NCS moiety is 103.5\u2005(4)\u00b0 and the bond angles for the coordinated atoms vary from 68.99\u2005(3)\u2013132.57\u2005(4)\u00b0, indicating a distorted geometry.The separation of the Mn atoms is 3.645\u2005(3) \u00c5. The Mn\u2014N(iso\u00adthio\u00adcyanato) and Mn\u2014N(azomethine) distances are 2.1289\u2005(11) and 2.3388\u2005(10)\u2005\u00c5 and the Mn\u2014O distances involving the coordinated water mol\u00adecules and \u03bc-N\u22efO2v [2.7971\u2005(13)\u2005\u00c5] hydrogen bond between adjacent dimers forms chains extending along the ac diagonal. The weak O4W\u2014H4W1\u22efS1iv inter\u00adaction [3.3159\u2005(12)\u2005\u00c5] and O2W\u2014H2W1\u22efO4W hydrogen bond [2.7322\u2005(14)\u2005\u00c5] link the dimers, generating a two-dimensional network as shown in Fig.\u00a02W, O3W and O4W are involved in O\u2014H\u22efO hydrogen-bonding inter\u00adactions , 3206 (b) [\u03bd(N\u2014H)], 2096 (s) [\u03bd(C\u2261N)], 1705 (s) [\u03bd [\u03bdasym (C=O)], 1555 (s) [\u03bd(C=N)], 1397 (s) [\u03bdsym(C=O)], 1067 (s) [\u03bd(N\u2014N)].Stoichiometric qu\u00adanti\u00adties of glyoxylic acid , ethyl\u00adcarbazate and ammonium thio\u00adcyanate were dissolved in 30\u2005mL of double-distilled water. To this homogeneous solution, Mn values were set to a multiple of Ueq(C) with 1.5 for CH3 and 1.2 for C\u2014H groups, respectively. Positions and Uiso values of water and amine H atoms were freely refined.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018014871/jj2203sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018014871/jj2203Isup2.hklStructure factors: contains datablock(s) I. DOI: 1870123CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The reaction of 2-methyl\u00adthio-5-amino-1,3,4-thia\u00addiazole with copper(II) acetate monohydrate resulted in the formation of the title binuclear compound 3H5N3S2) with copper(II) acetate monohydrate resulted in the formation of the title binuclear compound, [Cu2(C2H3O2)4(C3H5N3S2)2] or [Cu2(OAc)4(Me-SNTD)2]. The structure has triclinic (PR22(12) ring motif. The mol\u00adecules are further linked by C\u2014H\u22ef\u03c0 inter\u00adactions between the thia\u00addiazole rings and the methyl groups of the acetate units.The reaction of 2-methyl\u00adthio-5-amino-1,3,4-thia\u00addiazole (Me-SNTD; C The Cu\u2014O bond lengths range from 1.962\u2005(2) to 2.001\u2005(2)\u2005\u00c5 and the Cu\u2014N distance is 2.180\u2005(3)\u2005\u00c5. The Cu\u22efCu distance is 2.6727\u2005(6)\u2005\u00c5 and each metal atom exhibits a Jahn\u2013Teller-distorted octa\u00adhedral geometry. The observed Cu\u2014O2 bond length of 1.983\u2005(2)\u2005\u00c5 is longer than the Cu\u2014O1 distance of 1.962\u2005(2)\u2005\u00c5. The elongation of this Cu\u2014O distance may be due to the intra\u00admolecular N3\u2014H\u22efO2 hydrogen bond (Table\u00a01Each copper atom is displaced by 0.754\u2005(3)\u2005\u00c5 from the plane defined by basal-plane atoms O1, O2, O3 and O4 towards the nitro\u00adgen atom, N2. The Cu1d Table\u00a01. The coni hydrogen bond, forming a six-membered ring. The dimers are connected through an inter\u00admolecular N3\u2014H3\u22efO4ii hydrogen bond between the NH (Me-SNTD) and the carboxyl\u00adate groups, forming chains propagating parallel to [001]. The above-mentioned hydrogen bonds give rise to \u03c0 inter\u00adactions between the thia\u00addiazole rings and the acetate methyl groups generate a three-dimensional supra\u00admolecular framework s Table\u00a01. Additiork Fig.\u00a03.et al., 20162(OAc)4L2], where L is a ligand with an oxygen or nitro\u00adgen ligator atom, have been well explored. The structures of 2-methyl\u00adthio-5-amino-1,3,4-thia\u00addiazole and a complex of this mol\u00adecule with cadmium have been deposited in the CSD [XUVPEK and 2-methyl\u00adthio-5-amino-1,3,4-thia\u00addiazole were dissolved separately in a mixture of methanol-di\u00adchloro\u00admethane , mixed together and stirred for 1.5\u2005h. The green solid that precipitated was dissolved in methanol to form a green solution. Single crystals of the complex suitable for X-ray analysis were obtained by slow evaporation of the solution over a period of 10\u2005d.Cu(OAc)Uiso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019010272/cq2032sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019010272/cq2032Isup2.hklStructure factors: contains datablock(s) I. DOI: 1941461CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "This is correlated with the formation of carb\u00adoxy\u00adlic acid inversion dimers linked by pairwise O\u2014H\u22efO hydrogen bonds in the crystal of the title compound rather than an intra\u00admolecular O\u2014H\u22efO hydrogen bond.The \u2013COOH group of the title compound adopts a 11H8F3NO3, adopts a cis configuration across the \u2013C=C\u2013 double bond in the side chain and the dihedral angle between the phenyl ring and side chain is 47.35\u2005(1)\u00b0. The \u2013COOH group adopts a syn conformation (O=C\u2014O\u2014H = 0\u00b0), unlike the anti conformation observed in related maleamic acids. In the crystal, inversion dimers linked by pairs of O\u2014H\u22efO hydrogen bonds are connected via N\u2014H\u22efO hydrogen bonds and C\u2014H\u22efO inter\u00adactions into (100) sheets, which are cross-linked by another C\u2014H\u22efO inter\u00adaction to result in a three-dimensional network. The Hirshfeld surface fingerprint plots show that the highest contribution to surface contacts arises from O\u22efH/H\u22efO contacts (26.5%) followed by H\u22efF/F\u22efH (23.4%) and H\u22efH (17.3%).The title mol\u00adecule, C This disparity is a result of O\u2014Hc\u22efO=Ca intra\u00admolecular hydrogen bonds present in related structures and not observed in (I)The mol\u00adecule of (I)in Fig.\u00a01, similarvia pairwise O2\u2014H2O\u22efO3 hydrogen bonds chains chain in its own right chains runs down the b-axis direction, thereby cross-linking the sheets into a three-dimensional network.In the crystal of (I)ds Fig.\u00a02 forming ns Fig.\u00a02, resultins Fig.\u00a02. The N1\u2014ht Fig.\u00a02. In addiht Fig.\u00a02 forming dnorm surfaces and two-dimensional fingerprint plots (FP) were generated to further investigate the inter\u00admolecular inter\u00adactions in (I)et al., 2017N and H2O hydrogen atoms are due to their involvement as donors in stronger hydrogen bonds, while faint spots near H8 and H9 atoms are due to the weak C\u2014H\u22efO inter\u00adactions involving these atoms , F\u22efH/H\u22efF , H\u22efH , C\u22efH/H\u22efC , C\u22efF/F\u22efC and F\u22efF inter\u00adactions, with other contacts contributing the remaining 10.2%.In the Hirshfeld surface analysis, ms Fig.\u00a03. Analysims Fig.\u00a03 showed tN-(ar\u00adyl)-maleamic acids have been reported to date with varied substituents on the phenyl ring. Three of these, namely N-(phen\u00adyl)maleamic acid (CCDC refcode: LOSJUZ) maleamic acid (QUYJUQ) maleamic acid (PILVAI) anti O=C\u2014O\u2014H conform\u00adation and an intra\u00admolecular O\u2014H\u22efO hydrogen bond, as noted above. In LOSJUZ, adjacent mol\u00adecules are linked by N\u2014H\u22efO hydrogen bonds into a flat ribbon, while in QUYJUQ, N\u2014H\u22efO hydrogen bonds link the mol\u00adecules into zigzag chains propagating parallel to [001] and these chains are further linked into sheets by weak \u03c0\u2013\u03c0 inter\u00adactions. In the crystal structure of PILVAI, symmetry-related mol\u00adecules are linked by N\u2014H\u22efN hydrogen bonds, forming centrosymmetric amine\u2013amide dimers. The dimers are linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds and weak N\u2014H\u22ef\u03c0 and \u03c0\u2013\u03c0 inter\u00adactions into a three-dimensional network.Nineteen N-(2-methyl\u00adphen\u00adyl)maleamic acid set to 1.2Ueq(C). The oxygen- and nitro\u00adgen-bound H atoms were located from difference-Fourier maps and freely refined.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019006509/hb7816sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019006509/hb7816Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019006509/hb7816Isup3.cmlSupporting information file. DOI: 1914411CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "I centre within a S4 donor set. The three-dimensional mol\u00adecular packing is sustained by amine-N\u2014H\u22efS(thione), N\u2014H\u22efN(triazol\u00adyl) and N\u2014H\u22efN(thio\u00adcyanate) hydrogen bonds.The centrosymmetric binuclear complex features a tetra\u00adhedral Ag H-1,2,4-triazole-5(4H)-thione-\u03ba2S:S]bis\u00ad{(thio\u00adcyanato-\u03baS)silver(I)}, [Ag2(SCN)2(C2H3N3S)4], is generated by crystallographic inversion symmetry. The independent triazole-3-thione ligands employ the exocyclic-S atoms exclusively in coordination. One acts as a terminal S-ligand and the other in a bidentate (\u03bc2) bridging mode to provide a link between two AgI centres. Each AgI atom is also coordinated by a terminal S-bound thio\u00adcyanate ligand, resulting in a distorted AgS4 tetra\u00adhedral coordination geometry. An intra\u00admolecular N\u2014H\u22efS(thio\u00adcyanate) hydrogen bond is noted. In the crystal, amine-N\u2014H\u22efS(thione), N\u2014H\u22efN(triazol\u00adyl) and N\u2014H\u22efN(thio\u00adcyanate) hydrogen bonds give rise to a three-dimensional architecture. The packing is consolidated by triazolyl-C\u2014H\u22efS(thio\u00adcyanate), triazolyl-C\u2014H\u22efN(thiocyanate) and S\u22efS [3.2463\u2005(9)\u2005\u00c5] inter\u00adactions as well as face-to-face \u03c0\u2013\u03c0 stacking between the independent triazolyl rings [inter-centroid separation = 3.4444\u2005(15)\u2005\u00c5]. An analysis of the calculated Hirshfeld surfaces shows the three major contributors are due to N\u22efH/H\u22efN, S\u22efH/H\u22efS and C\u22efH/H\u22efC contacts, at 35.8, 19.4 and 12.7%, respectively; H\u22efH contacts contribute only 7.6% to the overall surface.The complete mol\u00adecule of the binuclear title complex, bis\u00ad[\u03bc-1 I complex, (I)H-1,2,4-triazole-5(4H-thione) and thio\u00adcyanate ligands has been synthesized and its crystal and mol\u00adecular structures determined as part of our on-going studies in this area , the heterocyclic ligand in (I)et al., 201812(SCN)]2 (I)P21/n and is disposed about a crystallographic centre of inversion. The HtrzSH mol\u00adecules only employ their exocyclic thione-sulfur atoms in coordination, there being no Ag\u22efN contacts of note. Each AgI atom is coordinated by a terminally bound HtrzSH mol\u00adecule and by two thione-sulfur atoms derived from two \u03bc2-bridging HtrzSH mol\u00adecules. The coordination of each AgI atom is completed by a terminal, S-bound thio\u00adcyanate anion. The geometry around the silver centre defined by the S4 donor set is distorted tetra\u00adhedral with the S\u2014Ag\u2014S bond angles spanning about 25\u00b0, i.e. from a narrow 91.60\u2005(2)\u00b0 for S1\u2014Ag\u2014S1i, being subtended by the bridging S1 atoms, to a wide 127.43\u2005(2)\u00b0 for S2\u2014Ag\u2014S3; symmetry operation (i): 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z. The Ag2S2 core has the shape of a distorted rhombus as the Ag\u2014S1 bond length of 2.5596\u2005(7)\u2005\u00c5 is significantly shorter than the Ag\u2014S1i bond of 2.8188\u2005(7)\u2005\u00c5. The Ag\u2014S bond lengths fall in two distinct classes, with the Ag\u2014S1b and Ag\u2014St bond lengths being similar and shorter than Ag\u2014S1ib b Table\u00a01. Despite2S2 core, with the dihedral angles between the core and the N1- and N4-rings being 88.99\u2005(11) and 85.16\u2005(11)\u00b0, respectively. The independent rings are close to being co-planar, exhibiting a dihedral angle of 8.38\u2005(16)\u00b0. Finally, the N1-amine is orientated to be in close proximity to the S3-thio\u00adcyanato atom, enabling the formation of an intra\u00admolecular amine-N\u2014H\u22efS(thio\u00adcyanato) hydrogen bond (Table\u00a02The five-membered rings lie prime to either side of the Agd Table\u00a02. While tii = 3.4444\u2005(15)\u2005\u00c5 and angle of inclination = 6.81\u2005(16)\u00b0 for (ii) x, y, z].The crystal of (I)et al., 2004et al., 2019Crystal Explorer 17 of the Hirshfeld surface plotted over dnorm for (I)i.e. S\u22efS contacts, are noted. The closest of these, i.e. S1\u22efS1iii = 3.2463\u2005(9)\u2005\u00c5 , link the binuclear mol\u00adecules into chains along the a-axis direction. On the Hirshfeld surface mapped over electrostatic potential (DFT 3-21G) shown in Fig.\u00a04b), the faint-red and light-blue regions correspond to negative and positive electrostatic potential, respectively,As seen in Fig.\u00a04a)\u2013(f), respectively. The N\u22efH/H\u22efN contacts, at 35.8%, are the major contributor to the Hirshfeld surface. The S\u22efH/H\u22efS contacts (19.4%) also make a significant contribution. Other significant contributions come from the C\u22efH/H\u22efC (12.7%) and S\u22efS (8.3%) contacts with H\u22efH contacts, occurring at distances beyond the sum of the van der Waals radii, contributing only 7.6%. The next most significant contribution is made by N\u22efC/C\u22efN contacts (6.7%) arising in the main from the \u03c0\u2013\u03c0 stacking inter\u00adactions between triazolyl rings.The full and delineated two-dimensional fingerprint plots are shown in Fig.\u00a05et al., 2017et al., 2017i.e. electrostatic (Eele), polarization (Epol), dispersion (Edis) and exchange\u2013repulsion (Erep); these were obtained using the wave function calculated at the HF/3-21G level of theory. The results are summarized in Table\u00a03\u22121, having a major electrostatic contribution (\u2212142.4\u2005kJ\u2005mol\u22121), and is associated with the following inter\u00adatomic contacts: C2\u2014H2\u22efN7, C4\u2014H4\u22efN7 and \u03c0\u2013\u03c0 stacking of between triazole rings. The next most significant contribution, with a total energy of \u2212125.0\u2005kJ\u2005mol\u22121, arises from conventional hydrogen bonds, i.e. N1\u2014H1N\u22efN5, N4\u2014H4N\u22efN2 and N6\u2014H6N\u22efN7 as well as C2\u2014H2\u22efS3 inter\u00adactions. The next attractive inter\u00adaction, with Etot = \u221248.9 and Edis = \u2212120.3\u2005kJ\u2005mol\u22121, respectively, reflects the N3\u2014H3N\u22efS2 hydrogen bonding and S1\u22efS1 secondary bonding contact.The energy frameworks were simulated \u2013(c), respectively, by energy framework diagrams whereby the cylinders join the centroids of mol\u00adecular pairs using a red, green and blue colour scheme; the radius of the cylinder is proportional to the magnitude of inter\u00adaction energy.The magnitudes of inter\u00admolecular energies, NWChem package supporting information and their energies were calculated to be 3.011 and 6.173\u2005eV, respectively. The HOMO is delocalized across the thio\u00adcyanato groups and the bridging region between the two dimers. The LUMO includes the delocalization around the triazole rings.The HOMO and LUMO energies for the atom positions in the crystal structure of (I)et al., 2016via the thione-S atom, as in (I)3P)2Cu(HtrzSH)Cl]\u00b7CH3CN, ]\u00b7CH3OH ]\u00b7H2O 2(HtrzS)(HtrzSH)\u00b70.5H2O 2(HtrzSH)2]\u00b72H2O}n 2Cl2]n Cl]n (HtrzSH)\u00b70.5H2O 3Sn(HtrzS) (NO3)OH2]n 2(SO4)]n H-1,2,4-triazole-3-thiol in distilled water (5\u2005ml) was added followed by heating for 4.3\u2005h during which time the precipitate slowly dissolved. The clear solution was filtered and kept to evaporate at ambient temperature. After a few days, colourless trapezoidal prisms of (I)in vacuo. M.p.: 413\u2013417\u2005K. IR : 2108 (s) (C\u2261N), 1479 (s) (C=N), 1248 (w) (C\u2013N), 1054 (m) (C\u2014S) + (C\u2014N).Silver nitrate and potassium thio\u00adcyanate were dissolved in acetro\u00adnitrile (25\u2005ml) and a white precipitate formed. This mixture was heated at 323\u2013325\u2005K for 30\u2005min. Then, a clear solution of 1Uiso(H) = 1.2Ueq. The maximum and minimum residual electron density peaks of 0.96 and 1.04 e\u2005\u00c5\u22123, respectively, were located 0.83 and 0.77\u2005\u00c5 from the N3 and Ag atoms, respectively.Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989019016359/hb7868sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019016359/hb7868Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019016359/hb7868sup3.tifFig. S1 Image of the calculated HOMO and LUMO's for (I). DOI: 1969873CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, two supra\u00admolecular tapes, each sustained by amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds and ten-membered {\u22efHNC 14H14N4O2\u00b7H2O, features a central C2N2O2 residue (r.m.s. deviation = 0.0205\u2005\u00c5) linked at each end to 3-pyridyl rings through methyl\u00adene groups. The pyridyl rings lie to the same side of the plane, i.e. have a syn-periplanar relationship, and form dihedral angles of 59.71\u2005(6) and 68.42\u2005(6)\u00b0 with the central plane. An almost orthogonal relationship between the pyridyl rings is indicated by the dihedral angle between them [87.86\u2005(5)\u00b0]. Owing to an anti disposition between the carbonyl-O atoms in the core, two intra\u00admolecular amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds are formed, each closing an S(5) loop. Supra\u00admolecular tapes are formed in the crystal via amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds and ten-membered {\u22efHNC2O}2 synthons. Two symmetry-related tapes are linked by a helical chain of hydrogen-bonded water mol\u00adecules via water-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonds. The resulting aggregate is parallel to the b-axis direction. Links between these, via methyl\u00adene-C\u2014H\u22efO(water) and methyl\u00adene-C\u2014H\u22ef\u03c0(pyrid\u00adyl) inter\u00adactions, give rise to a layer parallel to (10The mol\u00adecular structure of the title bis-pyridyl substituted di\u00adamide hydrate, C In the same way, complexation to metals may also be envisaged. It is therefore not surprising that there is now a wealth of structural information for these mol\u00adecules occurring in co-crystals, salts and metal complexes, as has been reviewed recently Having both amide and pyridyl functionality, bis\u00ad\u2005\u00c5 for N3 and 0.0321\u2005(11)\u2005\u00c5 for C8. The N1- and N3-pyridyl rings form dihedral angles of 59.71\u2005(6) and 68.42\u2005(6)\u00b0, respectively, with the central plane and lie to the same side of the plane, having a syn-periplanar relationship. The dihedral angle formed between the pyridyl rings is 87.86\u2005(5)\u00b0, indicating an almost edge-to-face relationship. The carbonyl-O atoms have an anti disposition enabling the formation of intra\u00admolecular amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds that close S(5) loops, Table\u00a01The mol\u00adecular structures of the two constituents comprising the crystallographic asymmetric unit of (I)via amide-N\u2014H\u22efO(carbon\u00adyl) hydrogen bonds leading to a sequence of inter-connected ten-membered {\u22efHNC2O}2 synthons. Two such tapes are connected by hydrogen bonds provided by the water mol\u00adecule of crystallization. Thus, alternating water mol\u00adecules in helical chains of hydrogen-bonded water mol\u00adecules, being aligned along the b-axis direction and propagated by 21 symmetry, connect to 3LH2via water-O\u2014H\u22efN(pyrid\u00adyl) hydrogen bonds to form the one-dimensional aggregate shown in Fig.\u00a02a). The presence of methyl\u00adene-C\u2014H\u22efO(water) and methyl\u00adene-C\u2014H\u22ef\u03c0(pyrid\u00adyl) contacts stabilizes a layer lying parallel to (10b).Significant conventional hydrogen bonding is noted in the crystal of (I)Crystal Explorer 17 dnorm mapping are electrostatic in nature as can be seen from the distinctive blue (electropositive) and red (electronegative) regions on the surface, albeit with varying intensity, Fig.\u00a04a) and (b). This indicates no charge complementarity consistent with the inter\u00adaction beings mainly dispersive in nature.To verify the nature of the aforementioned inter\u00adactions, the a), the overall fingerprint plot of (I)3LH2, with splitting of the spike in the inter\u00adnal region due to the formation of the O\u2014H\u22efN hydrogen bond, Fig.\u00a05e), suggesting a prominent role played by the water mol\u00adecule in influencing the overall contacts in (I)4LH2 in which the overall surface contacts for 4LH2 are not very much influenced by the benzene mol\u00adecule as demonstrated by the similar profiles for the solvate and individual 4LH2 mol\u00adecule , i.e. H\u22efH \u223c2.26\u2005\u00c5 [\u03a3(vdW) = 2.40\u2005\u00c5], O\u22efH/H\u22efO \u223c1.88\u2005\u00c5 [\u03a3(vdW) = 2.72\u2005\u00c5], C\u22efH/H\u22efC \u223c2.62\u2005\u00c5 [\u03a3(vdW) = 2.90\u2005\u00c5] and N\u22efH/H\u22efN \u223c2.50\u2005\u00c5 [\u03a3(vdW) = 2.75\u2005\u00c5].The qu\u00adanti\u00adfication of the close contacts to the Hirshfeld surface was performed through the analysis of the two-dimensional fingerprint plots for (I)3LH2 mol\u00adecule, the dominance of these contacts follows the order H\u22efH , C\u22efH/H\u22efC , O\u22efH/H\u22efO and N\u22efH/H\u22efN . While the aforementioned inter\u00adactions are almost evenly distributed between the inter\u00adnal and external contacts for (I)3LH2 are found to either to be inclined towards the inter\u00adnal or external contact region compared with (I)versus -H\u22efO- (9.5%) and -N\u22efH- (8.8%) versus -H\u22efN- (4.6%), respectively, Fig.\u00a05c)\u2013(e).As for the individual di + de 2.26\u2005\u00c5), O\u22efH/H\u22efO and H\u22efN . In particular, the second most dominant contacts are found to be heavily inclined toward -O\u22efH- (30.5%) as compared to -H\u22efO- (8.9%), presumably due to relatively large contact surface area.The hydrate mol\u00adecule exhibits a completely different fingerprint profile, which is dominated by three major contacts, namely H\u22efH 2 dimer connected by a ten-membered {\u22efHNC2O}2 synthon has the greatest Eint energy of \u221273.0\u2005kJ\u2005mol\u22121 which is comparable in energy to the classical eight-membered {\u22efHOCO}2 synthon , O1W\u2014H1W\u22efN1 (\u221228.6\u2005kJ\u2005mol\u22121), O1W\u2014H2W\u22efO1W (\u221226.2\u2005kJ\u2005mol\u22121), C7\u22efO1 (\u221220.7\u2005kJ\u2005mol\u22121), C5\u2014H5\u22efN4 (\u221213.0\u2005kJ\u2005mol\u22121) and C1\u2014H1\u22efO1W (\u221210.5\u2005kJ\u2005mol\u22121) inter\u00adactions. As expected, the N2\u2014H2N\u22efO1, N3\u2014H3N\u22efO2, O1W\u2014H1W\u22efN1 and O1W\u2014H2W\u22efO1W inter\u00adactions are associated with distinct electropositive and electronegative sites and therefore, are mainly governed by electrostatic forces, while the rest of the close contacts are dispersive in nature. The relatively stable nature of the C12\u2014H12\u22efC7 and C6\u2014H6A\u22efO2 inter\u00adactions as compared to the O1W\u2014H1W\u22efN1 and O1W\u2014H2W\u22efO1W inter\u00adactions could be due to the presence of low repulsion energies in the former as compared to the latter.All associations between mol\u00adecules in (I)N\u22efO1/ N3\u2014H3N\u22efO2, O1W\u2014H1W\u22efN1 and O1W\u2014H2W\u22efO1W hydrogen bonding leading to a barricade-like electrostatic energy framework parallel to (a). This is further stabilized by the dispersion forces arising from other supporting inter\u00adactions which result in another barricade-like dispersion energy framework parallel to (100), Fig.\u00a06b). The overall energy framework for (I)c).The crystal of (I)3LH2 mol\u00adecule in (I)i.e. Form I and Form II A comparison of the distribution of contacts on the Hirshfeld surfaces between the 3LH2 mol\u00adecule in (I)Mercury This conclusion is consistent with the analysis of the packing similarity in which a comparison of (I)3LH2 mol\u00adecule has been characterized in two polymorphs C(=O)N(H)CH2CO2H to 84.61\u2005(9)\u00b0. The comparable range for the S-shaped mol\u00adecules, for which both dihedral angles are identical from symmetry, is 64.2\u2005(3) to 84.79\u2005(18)\u00b0.The N,N\u2032-bis\u00ad(pyridin-3-ylmeth\u00adyl)oxalamide, was prepared according to the literature : 3578 \u03bd(O\u2014H), 3321 \u03bd(N\u2014H), 3141\u20132804 \u03bd(C\u2014H), 1687\u20131649 \u03bd(C=O), 1524\u20131482 \u03bd(C=C), 1426 \u03bd(C\u2014N), 710 \u03bd(C=C).The precursor, Uiso(H) set to 1.2\u20131.5Ueq(C). The oxygen- and nitro\u00adgen-bound H atoms were located in a difference-Fourier map and refined with O\u2014H = 0.84\u00b10.01\u2005\u00c5 and N\u2014H = 0.88\u00b10.01\u2005\u00c5, respectively, and with Uiso(H) set to 1.5Ueq(O) or 1.2Ueq(N). Owing to poor agreement, one reflection, i.e. (551), was omitted from the final cycles of refinement.Crystal data, data collection and structure refinement details are summarized in Table\u00a0610.1107/S2056989019016153/hb7869sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019016153/hb7869Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019016153/hb7869Isup3.cmlSupporting information file. DOI: 1969282CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "CO2 can be eliminated by renal replacement therapy but studies are scarce and clinical relevance is unknown. We prospectively studied CO2 and O2 behavior at different sample points of continuous veno-venous hemofiltration (CVVH) and build a model to calculate CO2 removal bedside.Carbon dioxide (CO2 (tCO2), CO2 flow (V\u0307CO2) and O2 flow (V\u0307O2) were compared between different sample points. The effect of citrate on transmembrane tCO2 was evaluated. Wilcoxon matched-pairs signed rank test was performed to evaluate significance of difference between 2 data sets. Friedman test was used when more data sets were compared.In 10 patients receiving standard CVVH under citrate anticoagulation, blood gas analysis was performed at different sample points within the CVVH circuit. Citrate was then replaced by NaCl 0.9% and sampling was repeated. Total CO2 in the effluent (26.0\u2009ml/min) correlated significantly with transmembrane V\u0307CO2 (24.2\u2009ml/min). This represents 14% of the average expired V\u0307CO2 in ventilated patients. Only 1.3\u2009ml/min CO2 was removed in the de-aeration chamber, suggesting that CO2 was almost entirely cleared across the membrane filter. tCO2 values in effluent, before, and after the filter were not statistically different. Transmembrane tCO2 under citrate or NaCl 0.9% predilution also did not differ significantly. No changes in V\u0307O2 were observed throughout the CVVH circuit. Based on recorded data, formulas were constructed that allow bedside evaluation of CVVH-attributable CO2 removal.V\u0307CO2 is removed by CVVH and can be quantified by one simple blood gas analysis within the circuit. Future studies should assess the clinical impact of this observation.A relevant amount of COhttps://clinicaltrials.gov with trial registration number NCT03314363 on October 192,017.The trial was registered at The online version of this article (10.1186/s12882-019-1378-y) contains supplementary material, which is available to authorized users. Subsequently, citrate predilution was stopped and replaced for at least 20\u2009min by NaCl 0.9% at similar flow. Blood gas analysis was repeated according to the same protocol.Inclusion and exclusion criteria are listed in Additional file\u00a02 content (tO2) was calculated as Hb x Hb saturation \u00d7\u20091.35 . V\u0307CO2 dropped significantly between SP2 and SP3 [from 110.5\u2009\u00b1\u20099.6\u2009ml/min to 84.5\u2009\u00b1\u20096.5\u2009ml/min (p\u2009<\u20090.01)]. V\u0307CO2 at SP4 (26.0\u2009\u00b1\u20095.8\u2009ml/min) and transmembrane V\u0307CO2 at SP4 (24.2\u2009\u00b1\u20092.6\u2009ml/min) were not statistically different (p\u2009=\u20090.39).V\u0307CO2 (SP1) than V\u0307CO2 (SP5) [109.8\u2009\u00b1\u20097.1\u2009ml/min vs. 81.7\u2009\u00b1\u20095.8\u2009ml/min (p\u2009<\u20090.01)] and a 1.3\u2009ml/min difference between V\u0307CO2 (SP3) and V\u0307CO2 (SP5) [83.0\u2009\u00b1\u20094.9\u2009ml/min vs. 81.7\u2009\u00b1\u20095.8\u2009ml/min (p\u2009=\u20090.01)].Results for NaCl 0.9% postdilution were plotted in Fig.\u00a02 at SP5 (116.1\u2009\u00b1\u20099.7\u2009ml/min) and \u201cexpected V\u0307CO2 at SP5\u201d (94.4\u2009\u00b1\u20099.9\u2009ml/min) , SP3 (25.0\u2009\u00b1\u20092.6\u2009mmol/l) and SP4 (25.1\u2009\u00b1\u20092.6\u2009mmol/l) were not statistically different (p\u2009=\u20090.51). tCO2 decreased significantly between SP1 and SP2 from 30.6\u2009\u00b1\u20092.3\u2009mmol/l to 25.5\u2009\u00b1\u20092.8\u2009mmol/l (p\u2009<\u20090.01). At all SP, tCO2 consisted of CO2 in gas form (pCO2) and HCO3- Fig.\u00a0.Fig. 6Ef2 at SP1, SP2, SP3 and SP5 was respectively 10.6\u2009\u00b1\u20093.7\u2009ml/min,10.9\u2009\u00b1\u20093.9\u2009ml/min, 10.3\u2009\u00b1\u20093.8\u2009ml/min, and 10.9\u2009\u00b1\u20093.7\u2009ml/min. V\u0307O2 at SP4 was 0\u2009ml/min as effluent contains no Hb. V\u0307O2 (SP1) and V\u0307O2 (SP5) were not different (p\u2009=\u20090.33) was statistically different, probably because the set CVVH fluid flow at these SP did not correspond with real fluid flow . By adding predilution fluid, tCO2 decreased between SP1 and SP2.As suggested by in vitro hemodialysis, COas HCO3- . The CO22 extraction. Only the short term effect of citrate upon CO2 removal was evaluated as an influencer of acid-base homeostasis. Over a longer time period, citrate could possibly affect CO2 clearance because it preserves membrane porosity better than heparin. tCO2 in blood passing through the CVVH circuit decreased as it was diluted by bicarbonate-free solutions. CVVH had no impact on V\u0307O2 because values remained constant at the different SP.Citrate anticoagulation did not influence tCO2 removal by CVVH in a clinical setting with the use of only one blood gas analysis in the extracorporeal circuit at a preexisting sample point. As these are the first data that were acquired in a CVVH setting, formulas could not be compared to data from other articles [Based on previous findings, different formulas were constructed to calculate COarticles . Further2 influences expired CO2.Several limitations of our study must be emphasized. First, despite the high number of analyses per patient, the sample size remains small and future studies in more patients are needed to confirm our results. Second, assumptions were made based on \u201csnapshot\u201d blood gas analysis. Continuous monitoring would be more precise. Third, fluid flows as set on CVVH may not correlate with real flow . In-circ2, both as gas and bicarbonate and measurable in the effluent, is removed during CVVH under citrate anticoagulation. Pre-filter tCO2 is the major determinant for CO2 removal. Citrate does not influence CO2 elimination. To a certain extent, bicarbonate fluids influence blood gases but data are too limited to permit relevant conclusions. Oxygen flow is not influenced by CVVH. CO2 removal by CVVH in bicarbonate-free conditions can be calculated by multiplying effluent or blood flow with CO2 content at a preexisting sample point. Their clinical relevance requires confirmation.A significant amount of COAdditional file\u00a01:Inclusion and exclusion criteria that were used during the study. (DOCX 15 kb)Additional file\u00a02:CVVH settings and postdilution fluid per patients. (DOCX 14 kb)"} +{"text": "The asymmetric unit of the title co-crystal comprises two twisted mol\u00adecules of 2,2\u2032-thiodi\u00adbenzoic acid and four mol\u00adecules of tri\u00adphenyl\u00adphosphane oxide. The three-dimensional mol\u00adecular packing is stabilized by hy\u00addroxy-O\u2014H\u22efO(oxide) hydrogen bonds and TPPO-C\u2014H\u22efO and TDBA-C\u2014H\u22ef inter\u00adactions. 14H10O4S\u00b72C18H15OP, comprises two mol\u00adecules of 2,2\u2032-thiodi\u00adbenzoic acid benzoic acid] and four mol\u00adecules of tri\u00adphenyl\u00adphosphane oxide . The two TDBA mol\u00adecules are twisted about their di\u00adsulfide bonds and exhibit dihedral angles of 74.40\u2005(5) and 72.58\u2005(5)\u00b0 between the planes through the two SC6H4 residues. The carb\u00adoxy\u00adlic acid groups are tilted out of the planes of the rings to which they are attached forming a range of CO2/C6 dihedral angles of 19.87\u2005(6)\u201360.43\u2005(8)\u00b0. Minor conformational changes are exhibited in the TPPO mol\u00adecules with the range of dihedral angles between phenyl rings being \u22122.1\u2005(1) to \u221262.8\u2005(1)\u00b0. In the mol\u00adecular packing, each TDBA acid mol\u00adecule bridges two TPPO mol\u00adecules via hy\u00addroxy-O\u2014H\u22efO(oxide) hydrogen bonds to form two three-mol\u00adecule aggregates. These are connected into a three-dimensional architecture by TPPO-C\u2014H\u22efO and TDBA-C\u2014H\u22ef inter\u00adactions. The importance of H\u22efH, O\u22efH/H\u22efO and C\u22efH/H\u22efC contacts to the calculated Hirshfeld surfaces has been demonstrated. In terms of individual mol\u00adecules, O\u22efH/H\u22efO contacts are more important for the TDBA (ca 28%) than for the TPPO mol\u00adecules (ca 13%), as expected from the chemical composition of these species. Computational chemistry indicates the four independent hy\u00addroxy-O\u2014H\u22efO(oxide) hydrogen bonds in the crystal impart about the same energy (ca 52\u2005kJ\u2005mol\u22121), with DTBA-phenyl-C\u2014H\u22efO(oxide) inter\u00adactions being next most stabilizing (ca 40\u2005kJ\u2005mol\u22121).The asymmetric unit of the title co-crystal, 2,2\u2032-thiodi\u00adbenzoic acid\u2013tri\u00adphenyl\u00adphosphane oxide (1/2), C In medicine, is dianion is found in the salt Na to 1.5018\u2005(8)\u2005\u00c5 [P1=O1].The mol\u00adecular structures of the TPPO coformers are more rigid. This is seen in the O\u2014P\u2014C\u2014C torsion angles, which range from 17.7\u2005(1) to 61.6\u2005(1), 19.8\u2005(1) to 61.5\u2005(1), \u22122.1\u2005(1) to \u221262.8\u2005(1) and \u221219.2\u2005(1) to \u221244.5\u2005(1)\u00b0 for the P1\u2013P4-mol\u00adecules, respectively. In the same way, the P=O bond lengths span an experimentally equivalent range, PLATON; Spek, 20093SC3OH\u22efO} heterosynthon as illustrated in Fig.\u00a04i.e. TPPO-C47\u2014H\u22efO11(carbon\u00adyl) and TPPO-C71\u2014H\u22efO5(carbon\u00adyl), operate in concert with hy\u00addroxy-O12\u2014H\u22efO3(oxide) and hy\u00addroxy-O6\u2014H\u22efO4(oxide) hydrogen bonds, respectively, to close a nine-membered {HC2PO\u22efHOCO\u22ef} synthon. The C\u2014H\u22efO contacts are of the type TPPO-C\u2014H\u22efO and TDBA-C\u2014H\u22efO, Table\u00a01i.e. the O5 and O11 atoms, participates in two C\u2014H\u22efO(carbon\u00adyl) inter\u00adactions, leaving no formal role for the carbonyl-O7 and O9 atoms in the mol\u00adecular packing. A view of the unit-cell contents is shown in Fig.\u00a05Geometric parameters characterizing the identified \u2013(f), the pair of TDBA-S1 and -S2 mol\u00adecules, shown with the respective pairs of hydrogen bonded TPPO mol\u00adecules, as well as the TPPO-P1\u2013P4 mol\u00adecules exhibit some similarities especially on the prominent close contacts as represented by the intense red regions on the corresponding dnorm surface mappings, which are mainly dominated by hydroxy-O\u2014H\u22efO(oxide) inter\u00adactions.The independent 2,2\u2032-thiodi\u00adbenzoic acid (TDBA) and tri\u00adphenyl\u00adphosphane oxide (TPPO) mol\u00adecules of (I)et al., 2008Upon close inspection on the surface mapping, minor differences are observed between the pair of TDBA mol\u00adecules. Specifically, a diminutive red spot is observed near one of the terminal carb\u00adoxy\u00adlic groups of the S1-mol\u00adecule arising from a TPPO-phenyl-C\u2014H\u22efO(carbonyl) inter\u00adaction but, no such contact is apparent for the S2-mol\u00adecule. As for the two pairs of TPPO mol\u00adecules, the significant difference between the TPPO-P1 and -P4 mol\u00adecules, linked to S1-DTBA, and the TPPO-P2 and P3 mol\u00adecules, linked to the S2-TDBA, is the presence of additional red spots on the surface mapping of the phenyl rings for P1- and P2-mol\u00adecules in contrast to their P3- and P4-containing counterparts. This difference may be attributed to the complementary phenyl-C\u2014H\u22ef\u03c0(phen\u00adyl) inter\u00adactions between centrosymmetrically-related mol\u00adecules, as illustrated in Fig.\u00a07di and de contact distances in inter\u00advals of 0.01\u2005\u00c5 gives the overall two-dimensional fingerprint plots for the entire asymmetric unit of (I)a), and each of the individual TDBA, Fig.\u00a09a), and TPPO, Fig.\u00a010a), mol\u00adecules. Further, these can be delineated into specific contacts \u2013(d) give fingerprint plots delineated into H\u22efH, O\u22efH/H\u22efO and C\u22efH/H\u22efC contacts. The relative contributions of these contacts to the surfaces is given in Table\u00a03Qu\u00adanti\u00adtative evaluation of the Hirshfeld surfaces by the combination of the a, is quite different for the individual components, Figs. 9a, as the former is a sum of all the individual surface contacts, which differ for the individual mol\u00adecules. As expected, the same is true for the corresponding decomposed fingerprint plots. The major contribution to the overall surface of (I)i.e. 49.4%, comes from H\u22efH contacts. The O\u22efH/H\u22efO contacts (de + di \u223c 2.34\u2005\u00c5) make a significant contribution at 13.7%, while the C\u22efH/H\u22efC inter\u00adactions (de + di \u223c 2.66\u2005\u00c5), at 30.1%, play a more prominent role.The overall fingerprint plot for (I)3SC3OH\u22efO} heterosynthon, Fig.\u00a04a), which exhibit an almost identical claw-like fingerprint profile but arranged in the exact reverse order, i.e. Fig.\u00a09a) cf. Fig.\u00a010a). Among all the close inter\u00adactions, H\u22efH contacts, Figs. 8b, represent the dominant inter\u00adactions to the individual surfaces, i.e. 41\u201342% for the TDBA mol\u00adecules and 49\u201351% for the DPPO mol\u00adecules, and exhibit de + di contact distances ranging from 2.24 to 2.38\u2005\u00c5 which is very close to the sum of van der Waals radii of 2.4\u2005\u00c5.The formation of the 13-membered {O\u22efHOCc). These feature two tips \u2013 one at relatively short de + di \u223c1.6\u2005\u00c5 that can be attributed to the hydroxy-H\u22efO(oxide) hydrogen bonds for the S1- and S2-TDBA mol\u00adecules, Fig.\u00a010c), or oxide-O\u22efH(hydrox\u00ady) hydrogen bonds for P1\u2013P4-TPPO. The other tip has a relatively long de + di value of \u223c2.4\u2005\u00c5 and arises as a result of hy\u00addroxy-O\u22efH(phen\u00adyl) contacts for S1- and S2-TDBA or phenyl-H\u22efO(hydrox\u00ady) for P1\u2013P4-TPPO. The O\u22efH/H\u22efO contacts constitute the second most dominant inter\u00adactions for the TDBA mol\u00adecules and third most for the TPPO mol\u00adecules, Table\u00a03The O\u22efH hydrogen bonds constitute the strongest among all inter\u00adactions present in the co-crystal and lead to formation of asymmetric, forceps-like profiles in the corresponding decomposed fingerprint plots, Figs. 9de + di distance range of 2.7\u20132.8\u2005\u00c5, i.e. close to the sum of van der Waals radii of 2.9\u2005\u00c5, despite the contacts constituting the third most dominant inter\u00adaction in the TDBA mol\u00adecules (ca 22%) and being the second most dominant for the TPPO mol\u00adecules (ca 32%). An exception to the trend is found for the P1- and P2-TPPO mol\u00adecules, which display relatively short contact distances at ca 2.6\u2005\u00c5 owing to the formation of C\u2014H\u22ef\u03c0 inter\u00adactions as discussed above.Similar to the H\u22efH contacts, the C\u22efH/H\u22efC inter\u00adactions contribute weakly to the mol\u00adecular packing of the co-crystal as evidenced from the et al., 2018In summary the Hirshfeld surface analysis on (I)d,p) available in Crystal Explorer of the hy\u00addroxy-O\u2014H\u22efO(oxide) hydrogen bonds is consistent across the series and lies in the range \u221250.7 to \u221253.3\u2005kJ\u2005mol\u22121. The other close contacts which exerts a relatively strong influence in the energy frameworks of the co-crystal are DTBA-phenyl-C\u2014H\u22efO(oxide) inter\u00adactions, with the Etot amounting of ca \u221240\u2005kJ\u2005mol\u22121, Table\u00a04Selected results obtained from the inter\u00adaction energy calculations involving the DTBA mol\u00adecules as reference mol\u00adecules are tabulated in Table\u00a04et al., 2005The only other structure of 2,2\u2032-thiodi\u00adbenzoic acid in the literature is that of the pure compound H. The observed P=O bond lengths in (I)i.e. in the range 1.4975\u2005(8) to 1.5018\u2005(8)\u2005\u00c5 are at the lower end of the range of such bonds.A survey of the Cambridge Structural Database : 3062 \u03bd(C\u2014H), 1693 \u03bd(COO), 1236 \u03bd(P=O), 1116 \u03bd(P\u2014Ar), 719 \u03b4(P\u2014C), 617 \u03bd(C\u2014S).All chemical precursors were of reagent grade and used as received without purification. Thio\u00adsalicylic acid and tri\u00adphenyl\u00adphosphane were dissolved in aceto\u00adnitrile (40\u2005ml) and the mixture subsequently added into an aceto\u00adnitrile solution (25\u2005ml) of copper(I) iodide . The reaction mixture was stirred for 1\u2005h at room temperature before the white product was filtered, washed with cold ethanol and dried Uiso(H) set to 1.2Ueq(C). The oxygen-bound H atoms were located from difference Fourier maps and refined without constraint. Owing to poor agreement, three reflections, i.e. global, I. DOI: 10.1107/S205698901801544X/hb7782Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901801544X/hb7782Isup3.cmlSupporting information file. DOI: 1876525CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The title mol\u00adecule adopts a helical structure, in which two 2,3\u2032-bipyridyl units are twisted up and down relative to the plane of the central benzene ring. Weak inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions lead to formation of a two-dimensional supra\u00admolecular network. Hirshfeld surface analysis indicates that the mol\u00adecular packing in the title compound is mainly dominated by inter\u00admolecular H\u22efH and H\u22efC/C\u22efH inter\u00adactions. 38H42N4O4, displays a helical structure induced by the combination of the C\u2014C\u2014C\u2014C torsion angle [\u221210.8\u2005(2)\u00b0] between two 2,3\u2032-bipyridyl units attached to the 1,2-positions of the central benzene ring and consecutive connections between five aromatic rings through the meta- and ortho-positions. Intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions between an H atom of a pyridine ring and the centroid of a another pyridine ring contributes to the stabilization of the helical structure. In the crystal, weak C\u2014H\u22ef\u03c0 inter\u00adactions link the title mol\u00adecules into a two-dimensional supra\u00admolecular network extending parallel to the ac plane, in which the mol\u00adecules with right- and left-handed helical structures are alternately arranged. Hirshfeld surface analysis and two-dimensional fingerprint plots indicate that the mol\u00adecular packing is dominated by van der Waals inter\u00adactions between neighbouring H atoms, as well as by C\u2014H\u22ef\u03c0 inter\u00adactions. One isopropoxyl group is disordered over two sets of sites [occupancy ratio 0.715\u2005(5):0.285\u2005(5)].The title mol\u00adecule, C An intra\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adaction between aromatic H3 and the centroid of the N3/C17\u2013C21 ring as well as C\u2014H\u22efN/O hydrogen bonds \u00b0, which is believed to reduce the steric hindrance between the two 2,3\u2032-bipyridyl units. In combination with this torsion angle, the consecutive connections of five aromatic rings in the title mol\u00adecule lead to a helical structure. The central benzene unit occupies A\u22efCg1i and between (meth\u00adyl)H37C\u22efCg2ii was found to be disordered over two sets of sites [occupancy ratio 0.715\u2005(5):0.285\u2005(5)].Crystal data, data collection and crystal structure refinement details are summarized in Table\u00a0410.1107/S2056989018013002/wm5462sup1.cifCrystal structure: contains datablock(s) I, New_Global_Publ_Block. DOI: 10.1107/S2056989018013002/wm5462Isup2.hklStructure factors: contains datablock(s) I. DOI: 1867774CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "It consists of octa\u00adhedral [Co(NH3)6]3+ cation (Co1 site symmetry 1), tetra\u00adhedral [ReVIIO4]\u2212 anions (Re1 site symmetry 1) and octa\u00adhedral [ReIVF6]2\u2212 anions ] tetroxidorhenate(VII) tetra\u00adkis\u00ad[hexa\u00adfluorido\u00adrhenate(IV)] hexa\u00adhydrate, arose unexpectedly due to possible contamination of the K 2ReF6 starting material with KReO4. It consists of octa\u00adhedral [Co(NH3)6]3+ cation (Co1 site symmetry 1), tetra\u00adhedral [ReVIIO4]\u2212 anions (Re site symmetry 1) and octa\u00adhedral [ReIVF6]2\u2212 anions , [Co(NH3)6]3+ octa\u00adhedral cations (mean Co\u2014N = 1.962\u2005\u00c5), and the [ReO4]\u2212 tetra\u00adhedral anion (mean Re\u2014O = 1.719\u2005\u00c5) are slightly distorted. A network of N\u2014H\u22efF hydrogen bonds consolidates the structure. The crystal studied was refined as a two-component twin.The title hydrated salt, tris\u00ad[hexa\u00adamminecobalt(III)] tetraoxidorhenate(VII) tetra\u00adkis\u00ad[hexa\u00adfluorido\u00adrhenate(IV)] hexa\u00adhydrate, arose unexpectedly due to possible contamination of the K VII is dominated by the tetra\u00adhedral perrhenate anion, [ReO4]\u2212 anions can be prepared in high yield by the reduction of a perrhenate starting material in the corresponding concentrated HX acid with AHF2 followed by an aqueous work-up salts as well as their X-ray single-crystal structures was recently reported , an unexpected mixed-valence rhenium(IV)\u2013rhenium(VII) salt arose in an effort to prepare [Co(NH3)6]2[ReF6]3 by metathesis from K2[ReF6] and Co(NH3)6Cl3 in water (353\u2005K). Yellow\u2013orange needle-like crystals of (I) were obtained within two hours by slow evaporation in water at room temperature. The crystals of (I) are air stable over short periods, but decompose to a black material after six months of storage at ambient temperature.In the process of exploring the coordination chemistry of hexa\u00adfluoro\u00adrhenate(IV) compounds, the title compound (I) 6]3+ cation, three distinct [ReF6]2\u2212 anions, one [ReO4]\u2212 anion, and two water mol\u00adecules of crystallization: these components are held together by electrostatic forces and hydrogen bonding. Site symmetries for the metal atoms are Co1: 1 (Wyckoff position 18f), Re1: 3 (Wyckoff position 6c), Re2: 1 (Wyckoff position 18f), Re3: a), and Re4: b).The structure of (I) Fig.\u00a01 is built3)6]3+ cation in (I) is slightly distorted; the average Co\u2014N bond length of 1.962\u2005\u00c5 is in agreement with the average Co\u2014N bond lengths of 1.963\u2005\u00c5 in [Co(NH3)6](ReO4)\u00b72H2O 6](TcO4)3 , the shortest Co\u22efCo and N\u22efN separations between nearby [Co(NH3)6]3+ cations are 7.035\u2005(1) and 4.473\u2005(1)\u2005\u00c5, respectively.The octa\u00adhedral [Co(NH4]\u2212 anion in (I), the average Re\u2014O bond length (1.719\u2005\u00c5) is in agreement with the average Re\u2014O bond length of 1.720\u2005\u00c5 in [Co(NH3)6](ReO4)\u00b72H2O the values of three Re\u2014O bond lengths, are slightly shorter than the fourth one [Re\u2014O1 = 1.748\u2005(14)\u2005\u00c5]. In (I), all O\u2014Re\u00ad\u2014O bond angles in the [ReO4]\u2212 anion are 109.5\u2005(3)\u00b0. However, in [Co(NH3)6](ReO4)\u00b72H2O, the [ReO4]\u2212 anion is slightly distorted by up to 2.7\u00b0 \u2005\u00c5 to 1.929\u2005(6)\u2005\u00c5. All the Re\u2014F bond lengths in the Re3- and Re4-centred anions are of equal distances of 1.952\u2005(6) and 1.950\u2005(6)\u2005\u00c5, respectively, by symmetry. Overall, the average Re\u2014F bond length (1.834\u2005\u00c5) in (I) is notably shorter than the average Re\u2014F bond length (1.951\u2005\u00c5) in A2[ReF6] salts previously studied and its component ions 6]3+) are shown in Fig.\u00a026]2\u2212 anions and the water mol\u00adecules (Table\u00a01A perspective view of the unit-cell plots for (s Table\u00a01.I) is the only reported hexa\u00adhalogenorhenate\u2013perrhenate structure containing both rhenium(IV) and rhenium(VII). It is noted that K2[ReF6] used for the preparation of (I) was not characterized before use and the presence of perrhenate in (I) may be due to the presence of K[ReO4] in the starting material. Efforts to isolate the technetium (Tc-99) derivative compound, [Co(NH3)6]3 [(Tc(vii)O4) (Tc(iv)F6)4] are in progress.To the best of our knowledge, was dissolved in 2\u2005ml of hot water (353\u2005K), and [Co(NH3)6]Cl3 dissolved in 1\u2005ml of \u2005H2O was added. The solution was allowed to evaporate slowly at room temperature and yellow-orange needle-like crystals of (I) were obtained within two hours. The compound was washed with H2O (3 \u00d7 1\u2005ml), followed by iso\u00adpropanol (3 \u00d7 1\u2005ml) and then diethyl ether (3 \u00d7 1\u2005ml). Single crystals of (I) were grown in H2O by slow evaporation at room temperature. Yield: ca 91%. The presence of perrhenate in (I) is probably due to the presence of K[ReO4] in the starting material (i.e. K2ReF6).KCrystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019009757/hb7830sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019009757/hb7830Isup2.hklStructure factors: contains datablock(s) I. DOI: 1939234CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The anions are disordered and modelled in two orientations. The major conformations have occupancies of 57, 59, 60 and 79%The asymmetric unit of the title compound contains two di\u00adaqua\u00adbis\u00ad(ethyl\u00adenedi\u00adamine)\u00adcopper(II) cations and four nitro\u00adbenzoate anions. These are connected into three-mol\u00adecule aggregates 2N,N\u2032)copper(II) bis\u00ad(2-nitro\u00adbenzoate), [Cu(C2H8N2)2(H2O)2](C7H4NO4)2, two di\u00adaqua\u00adbis\u00ad(ethyl\u00adenedi\u00adamine)\u00adcopper(II) cations and four nitro\u00adbenzoate anions are present in the asymmetric unit. All four anions are \u2018whole-mol\u00adecule\u2019 disordered over two sets of sites. The major components have refined occupancies of 0.572\u2005(13), 0.591\u2005(9), 0.601\u2005(9) and 0.794\u2005(10). The CuII ions exhibit slightly distorted octa\u00adhedral geometries. In the crystal, cations and anions are connected to each other via N\u2014H\u22efO and O\u2014H\u22efO hydrogen bonds, forming a two-dimensional network parallel to (200). The inter\u00admolecular contacts in the crystal were further analysed using Hirshfeld surface analysis, which indicates that the most significant contacts are O\u22efH/H\u22efO (42.9%), followed by H\u22efH (35.7%), C\u22efH/H\u22efC (14.2%), C\u22efC (2.9%), C\u22efO/O\u22efC (2.2%), N\u22efH/H\u22efN (0.9%) and N\u22efO/O\u22efN (0.3%).In the title compound, di\u00adaqua\u00adbis\u00ad(ethyl\u00adenedi\u00adamine-\u03ba Copper(II) carboxyl\u00adates have been used as single precursors for the preparation of copper(II) oxide nanoparticles di\u00adaqua\u00adcopper(II) cations and four 2-nitro\u00adbenzoate anions. In both cations, the CuII ion is coordinated by four N atoms of the ethyl\u00adenedi\u00adamine ligands which chelate the metal in the equatorial plane, and two axially coordinated water mol\u00adecules forming a slightly distorted octa\u00adhedral geometry. The Cu\u2014N bond lengths range from 1.991\u2005(6) and 2.050\u2005(5)\u2005\u00c5 (Table\u00a012(H2O)2]2+ cations and 2.564\u2005(5)\u2005\u00c5 for Cu1B] are noticeably longer than the equatorial Cu\u2014N distances to 1.251\u2005(15)\u2005\u00c5] in the nitro group are close to the values observed for related compounds reported in the literature and (C1A\u2013C6A) at . These inter\u00adactions consolidate the three-dimensional structure \u00adcopper(II) 2-nitro\u00adbenzoate under stirring. The precipitate was filtered, dried and dissolved in a hot methanol solution containing ethyl\u00adenedi\u00adamine under stirring. The mixture was filtered and single crystals were obtained after slow evaporation for one week.An aqueous solution of sodium 2-nitro\u00adbenzoate was added to an aqueous solution of CuSOUiso(H) = 1.2Ueq(C) for all C-bound H atoms. The N-bound H atoms were located in a difference-Fourier map and refined with N\u2014H = 0.91\u2005\u00c5, and with Uiso(H) = 1.5Ueq(N). The H atoms bonded to O atoms were located in a difference map and treated as part of a rigid group with oxygen as the pivot atom. All four anions are whole-mol\u00adecule disordered over two sets of sites. The major components have refined occupancies of 0.572\u2005(13), 0.591\u2005(9), 0.601\u2005(9) and 794\u2005(10). The major and minor components of disorder for each anion were constrained using the SAME command in SHELXL I. DOI: 10.1107/S2056989019016669/lh5937Isup2.hklStructure factors: contains datablock(s) I. DOI: 1909170CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II atom in the title compound has a distorted trigonal\u2013bipyramidal coordination environment defined by four N atoms from two bidentate 1,10-phenanthroline ligands and one oxygen atom from one-half of the monodentate N,N\u2032-diglycinate ligand. The dinuclear complex cations and the nitrate counter-anions as well as the solvate mol\u00adecules are linked by an intricate network of hydrogen bonds.The Cu 2(C12H10N2O6)(C12H8N2)4](NO3)2\u00b7C12H12N2O6\u00b78H2O, is composed of a CuII atom with a distorted trigonal\u2013bipyramidal coordination environment defined by four N atoms from two bidentate 1,10-phenanthroline ligands and one oxygen atom from one-half of the monodentate N,N\u2032-diglycinate anion. The asymmetric unit is completed by one-half of the N,N\u2032-diglycine solvent mol\u00adecule, which is located on a centre of inversion, by one nitrate counter-anion and four water mol\u00adecules. In the crystal, the cationic complexes are linked via inter\u00admolecular \u03c0\u2013\u03c0 stacking and through lone-pair\u22ef\u03c0 inter\u00adactions involving the N,N\u2032-diglycinate anion and the phenanthroline ligands. The N,N\u2032-diglycine solvent mol\u00adecule is involved in classical and non-classical hydrogen-bonding inter\u00adactions, as well as \u03c0\u2013\u03c0 stacking inter\u00adactions. The centroid-to-centroid distances between aromatic entities are in the range 3.5402\u2005(5)\u20134.3673\u2005(4)\u2005\u00c5. The crystal structure is stabilized by further C\u2014H\u22efO contacts as well as by O\u2014H\u22efO and N\u2014H\u22efO hydrogen bonds between water mol\u00adecules, the nitrate anions, the N,N\u2032-diglycinate ligands, N,N\u2032-diglycine solvent mol\u00adecules and phenanthroline ligands, giving rise to a supra\u00admolecular framework. A Hirshfeld surface analysis was carried out to qu\u00adantify these inter\u00adactions.The centrosymmetric binuclear complex cation of the title compound, [Cu In the two structures, the embedded N,N\u2032-diglycate mol\u00adecule links the cationic buildings blocks by numerous supra\u00admolecular inter\u00adactions.In our synthetic approach, we employ such systems as electron-deficient bidentate aromatic ring systems such as phenanthroline or bi\u00adpyridine in order to block parts of the metal cation coordination sphere. Thus, the alternative assembly process lies in the use of the offered different \u03c0-inter\u00adaction possibilities, N,N\u2032-di\u00adglycine moiety is a bis-monodentate bridging anionic ligand in its deprotonated form, as well as a solvent mol\u00adecule in its neutral form in one crystal structure. The structural investigation and description of the supra\u00admolecular network is confirmed and discussed with the aid of a Hirshfeld surface analysis diglycine solvent mol\u00adecule.In a continuation of this work, we have now synthesized and determined the structure of a novel copper(II) coordination compound where the 2(C12H8N2)4(C12H10N2O6)](NO3)2\u00b7(C12H12N2O6)\u00b78H2O, comprises two bidentate phenanthroline ligands and one bridging monodentate N,N\u2032-diglycinate ligand for each CuII atom, defining a distorted trigonal\u2013bipyramidal coordination sphere. A crystallographic centre of inversion is located at the centroid of the bridging N,N\u2032-diglycinate anion as well as the neutral and non-coordinating N,N\u2032-diglycine solvent mol\u00adecule. The asymmetric unit is completed by one non-coordinating nitrate counter-anion and four water mol\u00adecules , 2.119\u2005(2) and 2.111\u2005(3)\u2005\u00c5, and the axial positions by N1 and N3 with shorter bonds each of 1.974\u2005(3)\u2005\u00c5, respectively. The bond angle N1\u2014Cu\u2014N3 is 174.71\u2005(11)\u00b0. The sum of the bond angles O1\u2014Cu\u2014N2 [136.69\u2005(11)\u00b0], O1\u2014Cu\u2014N4 [103.88\u2005(12)\u00b0] and N2\u2014Cu\u2014N4 [118.90\u2005(10)\u00b0] in the equatorial plane amounts to 359.47\u00b0, indicating only slight distortions. Distances and angles within the distorted trigonal\u2013bipyramidal coordination sphere of the CuII ion are similar to those found in the literature diglycine solvent mol\u00adecule [C36\u2014O4 = 1.205\u2005(6) and C36\u2014O5 = 1.316\u2005(5)\u2005\u00c5]. The O1\u2014C30\u2014O2 angle of 123.1\u2005(4)\u00b0 in the carboxyl\u00adate group is slightly smaller than in the carb\u00adoxy\u00adlic group [O4\u2014C36\u2014O5 = 124.3\u2005(4)\u00b0]. In the coordinating N,N\u2032-diglycate ligand, the deviations of atoms defining the central benzamido entity from its least-squares plane are 0.040\u2005(4)\u2005\u00c5 (C28), \u22120.084\u2005(3)\u2005\u00c5 (O3), 0.245\u2005(4)\u2005\u00c5 (N5) and 0.404\u2005(4)\u2005\u00c5 (C29), while in the N,N\u2032-diglycine solvent they are \u22120.018\u2005(4)\u2005\u00c5 (C34) , 0.102\u2005(3)\u2005\u00c5 (O6), \u22120.192\u2005(4)\u2005\u00c5 (N6) and \u22120.257\u2005(4)\u2005\u00c5 (C35). The angle between the amide group and the carboxyl\u00adate group connected through the sp3-hybridized methyl\u00adene carbon atom (N5\u2014C29\u2014C30) is 113.6\u2005(3)\u00b0, and for the neutral solvent mol\u00adecule it is (N6\u2014C35\u2014C36) 112.1\u2005(3)\u00b0. The dihedral angle between the planar carboxyl\u00adate group (O1/C30/O2) and the aromatic synthon (C25\u201327/C25\u2032\u2013C27\u2032) of the ligand is 84.1\u2005(3)\u00b0 and thus smaller than the value found in the free solvent mol\u00adecule of the aromatic synthon (C31\u2013C33/C31\u2032\u2013C33\u2018) and the planar carboxyl\u00adate group (O4/C36/O5) at 88.9\u2005(3)\u00b0. The dihedral angle between the mean planes of the two bidentate phenanthroline ligands is 61.71\u2005(5)\u00b0; the corresponding value between phenanthroline (N1/C1\u2013C12/N2) and the coordin\u00adating carboxyl\u00adate group (O1/C30/O2) is 79.9\u2005(4)\u00b0 and between phenanthroline (N3/C13\u2013C24/N4) and the carboxyl\u00adate group is 82.5\u2005(3)\u00b0, respectively.The equatorial plane of the Cuvia O\u2014H\u22efO, C\u2014H\u22efO and partly via N\u2014H\u22efO hydrogen bonds with water solvent mol\u00adecules, the phenanthroline ligands and the metal-coordinating N,N\u2032-di\u00adglycin\u00adate ligands diglycine solvent mol\u00adecule, as well as between phenanthroline ligands and the metal-coordinating N,N\u2032-diglycinate ligand stack these components along the different axes \u2005\u00c5 between Cg2\u22efCg4, 3.6686\u2005(5)\u2005\u00c5 between Cg3\u22efCg4, and 3.5402\u2005(5)\u2005\u00c5 between Cg5\u22efCg5, where Cg1, Cg2Cg, 3Cg, 4 and Cg5 are the centroids defined by the ring atoms N1/C1\u2013C4/C12, C4\u2013C7/C11\u2013C12, N2/C7\u2013C11, C31\u2013C33/C31\u2032\u2013C33\u2032 and C16\u2013C19/C23\u2013C24, respectively. These distances are in expected ranges diglycinate ligand leads to a longer Cg6\u22efCg7 separation of 4.3673\u2005(4)\u2005\u00c5 \u00b0 and is slightly increased in comparison with the lines through C26/Cg7 and the centroids Cg6/Cg7 with a value of 16.99\u2005(5)\u00b0. Distances shown in Fig.\u00a04Cg7 and C15/Cg7 are 3.4440\u2005(4) and 3.676\u2005(6)\u2005\u00c5 and between H26/Cg7 and C26/Cg7 are 3.5049\u2005(4) and 3.713\u2005(5)\u2005\u00c5 with observed angles of 96.8\u2005(3)\u00b0 (C15\u2014H15\u22efCg7) and 95.4\u2005(3)\u00b0 (C16\u2014H26\u22efCg7), respectively. Besides the previously mentioned forces, a lone-pair\u22ef\u03c0 inter\u00adaction between the O3 atom of the carboxyl\u00adate group of the metal-coordinating N,N\u2032-diglycinate ligand and the Cg5 centroid of a phenanthroline ligand are observed and associated with a distance of 3.739\u2005(4)\u2005\u00c5. This value is similar to those found in the literature analysis diglycine solvent mol\u00adecule in the range \u22120.6806 to 1.9484 a.u. are shown in Fig.\u00a05de \u223c 1.19\u2005\u00c5/di \u223c 0.85\u2005\u00c5 and di \u223c 0.68\u2005\u00c5/de \u223c 1.02\u2005\u00c5 as well as de \u223c 1.05\u2005\u00c5/di \u223c 0.70\u2005\u00c5 and di \u223c 1.12\u2005\u00c5/de \u223c 0.78\u2005\u00c5, which comprise 27.9% and 42.2% of the total amount on the HS, respectively.The HS mapped over N,N\u2032-diglycine solvent mol\u00adecule involved in hydrogen bonding, the HS mapped over the electrostatic potentials were calculated using TONTO diglycine solvent mol\u00adecule as the second largest contribution within the HS. This high relevance for the HS is attributed to the high proportion of hydrogen atoms in the structure of these entities. The H\u22efN/N\u22efH contacts contribute 3.7% to the cationic complex and 3.4% to the solvent mol\u00adecule to the total HS, respectively. Short contacts of the solvent mol\u00adecule with a minor contribution to the lattice of O\u22efN/N\u22efO (0.4%) and O\u22efO (0.1%) are also observed. The contribution of the different \u03c0\u2013\u03c0 inter\u00adactions used for the stacking of the cationic complex subunits and the N,N\u2032-diglycine solvent mol\u00adecule along the different axes is also significant for both entities. Therefore, the close H\u22efC/C\u22efH (16.7%), C\u22efC (12.0%), O\u22efC/C\u22efO (2.1%) and N\u22efC/C\u22efN (1.2%) contacts of the cationic complex are assigned to C\u2014H\u22ef\u03c0 inter\u00adactions, \u03c0\u2013\u03c0 stacking (face-to-face) and lone-pair\u22ef\u03c0 inter\u00adactions of the carbonyl group and stacking between the phenanthroline ligands diglycine solvent mol\u00adecule, the close C\u22efC (11.1%), H\u22efC/C\u22efH (9.9%), N\u22efC/C\u22efN (1.8%) and O\u22efC/C\u22efO (1.2%) contacts are assigned to \u03c0\u2013\u03c0 stacking (face-to-face), C\u2014H\u22ef\u03c0 inter\u00adactions and stackings of the phenanthrolines and lone-pair\u22ef\u03c0 inter\u00adactions of the carbonyl group diglycinate ligand as well as the non-coordinating N,N\u2032-di\u00adglycine solvent mol\u00adecule in the HS mapped over the shape-index, which represent the face-to-face \u03c0\u2013\u03c0 stacking inter\u00adactions diglycine using SciFinder diglycine, and their structures show a number of non-classical inter\u00adactions diglycine, was prepared by the method of Cleaver & Pratt (1955v/v) mixture of water and methanol (50\u2005ml) and refluxed for 30 minutes. The mixture was allowed to cool to room temperature, and a previously prepared aqueous solution of copper acetate (1\u2005mmol) was slowly added under continuous stirring. Pale-blue block-shaped crystals of the title compound were obtained by slow evaporation at room temperature.The starting material, ratt 1955. Cesium Uiso(H) = 1.2Ueq(C) and C\u2014H(aromatic) = 0.94\u2005\u00c5 and C\u2014H(methyl\u00adene) = 0.98\u2005\u00c5 using a riding model. The water H atoms were located in a difference-Fourier map and were refined with O\u2014H distances restrained to 0.82\u20130.87\u2005\u00c5 and with Uiso(H) = 1.5Ueq(O), except O11\u2014H11A with a fixed distance of 1.00\u2005\u00c5, which led to a stable and consolidated hydrogen-bonding network.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019005164/wm5501sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019005164/wm5501Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019005164/wm5501Isup3.cdxSupporting information file. DOI: 1910262CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal, the mol\u00adecules are linked by weak C\u2013H\u22efF hydrogen bonds into the supra\u00admolecular inversion dimers. 13H6Cl3FOS, the aromatic rings are inclined to one another by 12.9\u2005(2)\u00b0, and the thio\u00adphene ring is affected by \u03c0-conjugation. In the crystal, mol\u00adecules are linked by C\u2014H\u22efF hydrogen bonds, forming an R22(8) ring motif. A Hirshfeld surface analysis was conducted to verify the contribution of the different inter\u00admolecular inter\u00adactions. The shape-index surface clearly shows that the two sides of the mol\u00adecules are involved in the same contacts with neighbouring mol\u00adecules and the curvedness plots show flat surface patches characteristic of planar stacking.In the title chalcone\u2013thio\u00adphene derivative, C Chalcones, considered to be the precursor of flavonoids and isoflavonoids, are abundant in edible plants. Compounds with the 1,3-di\u00adphenyl\u00adprop-2-en-1-one framework are described by its generic term \u2018chalcone\u2019. They consist of open-chain flavonoids in which the two aromatic rings are joined by a three-carbon \u03b1,\u03b2-unsaturated carbonyl system. These are coloured compounds because of the presence of the \u2013CO\u2014CH=CH\u2013 chromophore, which depends in the presence of other auxochromes. Accumulating evidence has shown that chalcones and their derivatives could inhibit tumor initiation and progression. In view of the above, and as a part of our ongoing research on chalcone derivatives \u00b0 for the atoms O1\u2014C5\u2014C6\u2014C7. The thio\u00adphene ring is affected by \u03c0 conjugation. This can be explained by the longer C=S values of 1.703\u2005(6) and 1.714\u2005(4)\u2005\u00c5 for S1=C2 and S1=C1, respectively. The bond-angle values O1\u2014C5\u2014C6 [121.9\u2005(4)\u00b0], O1\u2014C5\u2014C4 [118.2\u2005(4)\u00b0] and C5\u2014C6\u2014C7 = 125.14\u2005(4)\u00b0 about C5 indicate that the carbon atom is in a distorted trigonal\u2013planar configuration, which is due to steric hindrance of the oxygen atom. The mol\u00adecular structure is stabilized by an intra\u00admolecular C6\u2014-H6A\u22efCl1 hydrogen bond (Table\u00a01S(6) motif, as shown in Fig.\u00a01The mol\u00adecular structure of the title compound, shown in Fig.\u00a01d Table\u00a01 that cloCg1\u22efCg1 = 3.956\u2005(3)\u2005\u00c5 and Cg2\u22efCg2 = 3.957\u2005(3)\u2005\u00c5 where Cg1 and Cg2 are the centroids of the S1/C1\u2013C4 and C8\u2013C13 rings, respectively.In the crystal, the mol\u00adecules are linked by C\u2014H\u22efF hydrogen bonds, forming an et al., 2016E)-3-(phen\u00adyl)-1-prop-2-en-1-one as the main skeleton revealed the presence of three structures containing a similar 2,5-di\u00adchloro\u00adthio\u00adphene\u2013chalcone moiety to the title compound but with different substituents on the terminal phenyl rings, viz. [(E)-1--3-(X)prop-2-en-1-one], where X = 4-(di\u00admethyl\u00adamino)\u00adphenyl and shape-index (\u22121.0 to 1.0 a.u.), respectively. The calculated volume inside the Hirshfeld surface is 325.37\u2005\u00c53 in the area of 310.17\u2005\u00c53.Hirshfeld surfaces and fingerprint plots were generated for the title compound based on the crystallographic information file (CIF) using A\u22efCl1 and C10\u2014H10A\u22efF1 inter\u00adactions, which play a significant role in the mol\u00adecular packing of the title compound. The Hirshfeld surfaces illustrated in Fig.\u00a04In Fig.\u00a04The overall two-dimensional fingerprint plot for the title compound and those delineated into Cl\u22efH/H\u22efCl, C\u22efC, Cl\u22efCl, Cl\u22efS/S\u22efCl, H\u22efH, F\u22efH/H\u22efF, C\u22efH/H\u22efC contacts are illustrated in Fig.\u00a05et al., 2013abet al., 2014et al., 2010bThe title compound was synthesized as per the procedure reported earlier (Kumar Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018010216/xu5930sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989018010216/xu5930Isup2.hklStructure factors: contains datablock(s) I. DOI: 1036795CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Correction to: J Cardiovasc Magnhttps://doi.org/10.1186/s12968-019-0537-4In the original version of this article , publishGadoteriol) application (T1 native: 5\u2009s(3\u2009s)3\u2009s; T1 post-contrast: 4\u2009s(1\u2009s)3\u2009s(1\u2009s)2\u2009s; TE 1.1\u2009ms and slice thickness 6\u2009mm).A sampling protocol with reduced sensitivity to heart rate was applied [15] before and 15\u2009min after contrast media application (T1 native: 5\u2009s(3\u2009s)3\u2009s; T1 post-contrast: 4\u2009s(1\u2009s)3\u2009s(1\u2009s)2\u2009s; TE 1.1\u2009ms and slice thickness 6\u2009mm).A sampling protocol with reduced sensitivity to heart rate was applied [15] before and 15\u2009min after contrast media (0.15\u2009mmol/kg body weight The corrected information is also repeated below:"} +{"text": "In the crystal, the only short inter\u00admolecular contacts are Cl\u22efO contacts [3.173\u2005(3)\u2005\u00c5] that link the mol\u00adecules to form a 21 helix propagating along the b-axis direction.In the title chalcone derivative, C 15H9BrCl2O, the aryl rings are inclined to each by 14.49\u2005(17)\u00b0, and the configuration about the C=C bond is E. There is a short intra\u00admolecular C\u2014H\u22efCl contact present resulting in the formation of an S(6) ring motif. In the crystal, the shortest inter\u00admolecular contacts are Cl\u22efO contacts [3.173\u2005(3)\u2005\u00c5] that link the mol\u00adecules to form a 21 helix propagating along the b-axis direction. The helices stack up the short crystallographic a axis, and are linked by offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.983\u2005(1)\u2005\u00c5], forming layers lying parallel to the ab plane. A qu\u00adanti\u00adfication of the inter\u00admolecular contacts in the crystal were estimated using Hirshfeld surface analysis and two-dimensional fingerprint plots.In the title chalcone derivative, C Chemically they consist of open-chain flavonoids in which the two aromatic rings are joined by a three-carbon, \u03b1-unsaturated carbonyl system and are described by the generic term \u2018chalcone\u2019. Chalcones are coloured compounds because of the presence of the \u2013CO\u2014CH=CH\u2013 chromophore, which depends on the presence of other auxochromes. Chalcones are finding applications as organic non-linear optical materials (NLO) because of their good SHG conversion efficiencies (Chandra Shekhara Shetty E configuration. There is a short intra\u00admolecular C\u2014H\u22efCl contact present resulting in the formation of an S(6) ring motif \u00b0. The trans conformation of the C=C double bond in the central enone group is confirmed by the C6\u2014C7\u2014C8=C9 torsion angle of \u2212179.8\u2005(3)\u00b0. The bond angles O1\u2014C9\u2014C10 [120.4\u2005(3)\u00b0], O1\u2014C9\u2014C8 [119.9\u2005(3)\u00b0] and C9\u2014C8\u2014C7 [123.9\u2005(4)\u00b0] about C9 indicate that this carbon atom is in a distorted trigonal\u2013planar conformation.The mol\u00adecular structure of the title compound is shown in Fig.\u00a01if Fig.\u00a01. The unsa axis. The shortest inter\u00admolecular contacts are Cl\u22efO1i contacts .In the crystal, the mol\u00adecules stack along the short crystallographic on Fig.\u00a02. The helCrystalExplorer shows an extensive flat surface characteristic of planar stacking \u2013 see the Supra\u00admolecular features section above.Hirshfeld surfaces and fingerprint plots were generated for the title compound using d Figs. 2 and 3 \u25b8,et al., 2007The overall two-dimensional fingerprint plot , including 1-(3-bromo\u00adphen\u00adyl)-3-phenyl\u00adprop-2-en-1-one itself \u00b0 because of the presence of the intra\u00admolecular C\u2014H\u22efCl hydrogen bond, as shown in Fig.\u00a01A search of the Cambridge Structural Database = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S205698901900104X/qm2132sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S205698901900104X/qm2132Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901900104X/qm2132Isup3.cmlSupporting information file. DOI: 10.1107/S205698901900104X/qm2132sup4.pdfDetails of CSD search. DOI: 1036741CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "One showcases a novel (dppm)C(N2dppm) PCP pincer, the other contains a (dppm)C(N2) diazo\u00admethyl\u00adene\u00adphospho\u00adrane moiety.In this communication, two compounds and their respective crystal structures, obtained 2(C58H51N3P4)]Cl\u00b75.5CH3CN or Cl\u00b75.5CH3CN , resulting from an oxygen-mediated cleavage of a triazeneyl\u00adidene\u00adphospho\u00adrane ligand producing a diazo\u00admethyl\u00adene\u00adphospho\u00adrane and a nitrene moiety, which in turn rearrange via a Staudinger reaction and a 1,2-hydride shift to the first title complex, involves a six-coordinate IrIII complex cation coordinated by a facial PCP pincer ligand, a benzaldimine and two chlorido ligands. The pincer system features a five- and a seven-membered ring, with the central divalent carbon of the PCP pincer ligand being connected to a phosphine and a diazo\u00adphospho\u00adrane. The chlorido ligands are positioned trans to the central carbon atom and to the phospho\u00adrus donor of the seven-membered ring of the pincer system, respectively. A chloride ion serves as counter-ion for the monocationic complex. The structure of [IrI(C26H22N2P2)(C26H22P2)(C6H7N)]I(I3)\u00b70.5I2\u00b7CH3OH\u00b70.5CH2Cl2 or I(I3)\u00b70.5I2\u00b7CH3OH\u00b70.5CH2Cl2 {4, systematic name: iodido(phenyl\u00admethanimine-\u03baN)iridium(III) iodide\u2013triiodide\u2013di\u00adchloro\u00admethane\u2013iodine\u2013methanol (2/2/1/1/2)}, accessed via treatment of the triazeneyl\u00adidene\u00adphospho\u00adrane complex Cl with hydro\u00adiodic acid, consists of a dicationic six-coordinate IrIII complex, coordinated by a bidentate diazo\u00admethyl\u00adene\u00adphospho\u00adrane, a benzaldimine, a chelating dppm moiety and an iodido ligand. The phospho\u00adrus atoms of the chelating dppm are trans to the central carbon atom of the diazo\u00admethyl\u00adene\u00adphospho\u00adrane and the iodide ligand, respectively. Both an iodide and a triiodide moiety function as counter-ions. The aceto\u00adnitrile solvent mol\u00adecules in 3 are severely disordered in position and occupation. In 4, the I3\u2212 anion is positionally disordered (ratio roughly 1:1), as is the I\u2212 anion with a ratio of 9:1. The di\u00adchloro\u00admethane solvent mol\u00adecule lies near a twofold rotation axis (disorder) and was refined with an occupancy of 0.5. Another disorder occurs for the solvent methanol with a 1:1 ratio.The structure of [IrCl This substitution reaction results in the formation of a labile IrI inter\u00admediate, whose coordination sphere features the PCN pincer ligand (4-Cl-C6H4N3)C(dppm) and a monodentate dppm are created by using benzyl azide, rather than 1-azido-4-chloro\u00adbenzene, under an inert atmosphere.In this contribution, we describe the fragmentation of a triazene into diazo and nitrene parts in the coordination sphere of iridium. Recently, we reported on the synthesis of Cl (2) with hydro\u00adiodic acid. It is apparent that a rupture of the N1\u2014N2 bond (numbering as in the structure of 4) of (BnN3)C(dppm) occurred again, resulting in the formation of a diazo\u00admethyl\u00adene\u00adphospho\u00adrane (dppm)C(N2) and a benzyl\u00adnitrene part. However, in this case, the diazo functionality remains unchanged, since in contrast to the formation of 3, no free phosphine functionality is available. The benzyl\u00adnitrene unit again undergoes a 1,2-hydride shift and, as a benzaldimine, coordinates to the Ir metal center.In a related fragmentation reaction, compound 2) moiety contains a central divalent carbon 2)(N2), X = Cl, Br, were obtained by addition of CX4 to P((NMe2)2)(CH(N2)) C ring and one five-membered IrC(dppm) ring. A benzaldimine ligand is positioned trans to the phospho\u00adrus donor of the five-membered ring, the remaining two coordination sites being occupied by chlorido ligands cis to each other. The deviations of the angles C1\u2014Ir1\u2014Cl1 = 170.06\u2005(13)\u00b0 and N3\u2014Ir1\u2014P1 = 169.02\u2005(11)\u00b0 from a regular octa\u00adhedral geometry indicate some strain in the pincer system. Both the N1\u2014C1 bond length [1.280\u2005(5)\u2005\u00c5] and the N1\u2014N2 bond length [1.445\u2005(5)\u2005\u00c5] are typical for a C=N double bond and an N\u2014N single bond, respectively. The P3\u2014N2 bond length [1.586\u2005(4)\u2005\u00c5] is in the range of P=N double bonds observed for imino\u00adphospho\u00adranes \u2005\u00c5] and a previously reported iridium benzaldimine complex [1.260\u2005(6)\u2005\u00c5] involving a phospho\u00adrus donor atom trans to the benzaldimine nitro\u00adgen donor \u2005\u00c5, D\u2014H\u22efA 138\u2005(4)\u00b0], while other intra\u00admolecular inter\u00adactions involve atoms N1 and Cl1 and the various phenyl rings , which forms a five-membered chelate ring via one C and one P donor atom. A four-membered ring is formed by a bidentate dppm ligand and is oriented perpendicular to the plane of the five-membered ring with one phospho\u00adrus donor trans to the carbon donor of the five-membered ring. The benzaldimine ligand is located trans to the phospho\u00adrus donor of the five-membered ring, the sixth coordination site is occupied by an iodido ligand. Deviations from the octa\u00adhedral symmetry around the Ir center are mainly due to the strained four-membered ring [P4\u2014Ir1\u2014P3 = 70.80\u2005(5)\u00b0] with consequences for the bond angles P4\u2014Ir1\u2013I1 [165.12\u2005(4)\u00b0] and C1\u2014Ir1\u2014P3 [170.90\u2005(17)\u00b0]. The C1\u2014Ir1\u2014P1 bond angle of the five-membered ring is 84.37\u2005(17)\u00b0. The environment around the ylidic carbon C1 is trigonal planar, with the bond angle N3\u2014C1\u2014P2 exhibiting the largest deviation from a regular symmetry [114.8\u2005(5)\u00b0]. Both the N3\u2014N2 [1.095\u2005(9)\u2005\u00c5] and the N3\u2014C1 [1.305\u2005(9)\u2005\u00c5] bond lengths are slightly shorter, compared to the corresponding mean values of ten previously reported structures of diazo compounds 2 and [CH(dppm)2]Cl , dissolved in acetone (0.6\u2005ml) and stirred for 3\u2005h. The resulting white precipitate of [IrCl2H(C(dppm)2)] were added. After stirring for 1\u2005min, the deep-purple solution was stirred for 2\u2005h under atmospheric conditions, resulting in the slow precipitation of a white product. Colourless to pale-yellow prismatic crystals of 3 were obtained by allowing the purple inter\u00admediate solution to stand overnight under ambient conditions.31P{1H}-NMR (CHCl3/MeOH 1:1): \u03b4 = 0.7 ; 16.9 ; 6.9 ; \u221243.1 ppm. 13C{1H}-NMR (CHCl3/MeOH 1:1): \u03b4 =157.9 ppm.Synthesis and crystallization of complex 4: Under an inert atmosphere, a mixture of [IrCl(cod)]2 , [CH(dppm)2]Cl were added. After heating to 333\u2005K for 15\u2005min, the volatiles were removed in vacuo. The residue was dissolved in CH2Cl2 and hydro\u00adiodic acid was added whilst stirring. The orange\u2013brown precipitate that formed slowly was separated, washed with water and dried in vacuo. A solution of the residue in CH2Cl2/MeOH 2:1 (0.6\u2005ml) qu\u00adanti\u00adtatively contained an unidentified inter\u00admediate, which transformed to the product within 1\u2005h. Red prismatic crystals of 4 formed within a few hours, when a solution of the inter\u00admediate in CH2Cl2/MeOH (5:1) was kept at 254\u2005K for 24\u2005h and subsequently warmed to room temperature.31P{1H}-NMR (CH2Cl2/MeOH 2:1): \u03b4 = \u221215.8 ; 37.8 ; \u221272.1 ; \u221262.8 ppm. 13C{1H}-NMR (CH2Cl2/MeOH 2:1): \u03b4 = \u22123.0 ppm.3 are severely disordered in position and occupation. At least 5.5 mol\u00adecules in the asymmetric units were refined. Occupation values were varied to give a reasonable isotropic displacement factor. All C- and N-atoms of solvent mol\u00adecules were refined isotropically with bond restraints, the hydrogen atoms were omitted. The proton on N3 was freely refined.Crystal data, data collection and structure refinement details are summarized in Table\u00a044 was found and refined with a bond restraint of 0.87\u2005(2)\u2005\u00c5. The I3\u2212 anion (I4\u2013I6) is positionally disordered (ratio roughly 1:1), as is the I\u2212 anion with a ratio I2:I2A of 9:1. The di\u00adchloro\u00admethane solvent mol\u00adecule lies near a twofold rotation axis (disorder) and was refined with an occupancy of 0.5. Another disorder occurs for the solvent methanol with a ratio of 1:1. The C and O atoms of methanol were refined isotropically with bond restraints of 1.40\u2005\u00c5. The hydrogen atoms of methanol were calculated, those of di\u00adchloro\u00admethane omitted. All other H atoms were positioned geometrically (C\u2014H = 0.94\u20130.98\u2005\u00c5) and refined as riding with Uiso(H) = 1.2-1.5 Ueq(C).The hydrogen atom at N1 of 10.1107/S2056989019000136/su5470sup1.cifCrystal structure: contains datablock(s) global, 3, 4. DOI: 10.1107/S2056989019000136/su54703sup2.hklStructure factors: contains datablock(s) 3. DOI: 10.1107/S2056989019000136/su54704sup3.hklStructure factors: contains datablock(s) 4. DOI: 1885936, 1885935CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "II complex, the axial positions are occupied by a nitro\u00adgen atom from the ligand and an oxygen atom from the sulfate anion, whereas in the CdII complex they contain two nitro\u00adgen atoms from two different ligands and the sulfate anion only serves as the charge-balancing ion. \u03c0\u2013\u03c0 stacking between pyridine rings plays a crucial role in the mol\u00adecular self-assembly of the two structures.The title compounds both contain a central metal atom in a distorted octa\u00adhedral geometry coordinated equatorially by four oxygen atoms from water mol\u00adecules. In the Mn N,N\u2032-bis\u00ad(pyridin-4-yl)pyridine-2,6-dicar\u00adboxamide]\u00adsulfatomanganese(II) dihydrate, [Mn(SO4)(C17H13N5O2)(H2O)4]\u00b72H2O or [Mn(H2L1)(SO4)(H2O)4]\u00b72H2O, (I), and tetra\u00adaqua\u00adbiscadmium(II) sulfate tetra\u00adhydrate, [Cd(C17H13N5O2)2(H2O)4]SO4\u00b74H2O or [Cd(H2O)4(H2L1)2]\u00b7SO4\u00b74H2O, (II), both contain a central metal atom in a distorted octa\u00adhedral geometry coordinated equatorially by four oxygen atoms from water mol\u00adecules. In (I), the axial positions are occupied by a nitro\u00adgen atom from H2L1 and an oxygen atom from the sulfate anion, whereas in (II), the axial positions contain two nitro\u00adgen atoms from two different H2L1 ligands and the sulfate anion acts as the charge-balancing ion. \u03c0\u2013\u03c0 stacking between pyridine rings and a network of hydrogen bonds involving the water molecules and the sulfate anions play a crucial role in the mol\u00adecular self-assembly of the two structures.The molecular structures of tetra\u00adaqua\u00ad[ This bonding feature, when combined with the flexibility and length of mol\u00adecular backbone within these ligands, can lead to the construction of porous coordination compounds -1H-1,2,3-triazol-1-yl] acetic acid (SO4)(H2O)4]\u00b72H2O, (I)2L1)2(H2O)4]SO4\u00b74H2O, (II)N,N\u2032-bis\u00ad(pyridin-4-yl)pyridine-2,6-dicarboxamide (H2L1) as the heterocyclic nitro\u00adgen ligand.In recent years, the design of metal\u2013organic complexes constructed from heterocyclic nitro\u00adgen-derivative ligands has witnessed an upsurge in inter\u00adest due to their fascinating structures and potential applications in luminescence, catal\u00adysis, gas storage and separation (Perry 2L1) acts as a monodentate ligand. The results indicate that the rational design and selection of ligands with heterocyclic nitro\u00adgen systems is an effective synthetic strategy to construct complexes via self-assembly. The asymmetry of the [N,N\u2032-bis\u00ad(pyridin-4-yl)pyridine-2,6-dicarboxamide ligand has resulted in some novel structures PII ion exhibits an octa\u00adhedral geometry, arising from coordination to five water and one sulfate oxygen atoms and to one nitro\u00adgen (N1) atom of a pyridine group of the H2L1 ligand (Table\u00a01II ion vary from 85.79\u2005(7) to 177.74\u2005(6)\u00b0. Intra\u00admolecular O4\u2014H4\u22efO8, N2\u2014H20\u22efN5 and N3\u2014H24\u22efN5 hydrogen bonds are also present (Table\u00a03The mononuclear complex (I)d Table\u00a01. The Mn\u2014t Table\u00a03.4\u00b7H2O is replaced by CdSO4\u00b78/3H2O, complex (II)C2/c. As illustrated in Fig.\u00a02II also shows an octahedral environment coordinating four oxygen atoms from four water molecules and two axial nitrogen atoms from two symmetry-related H2L1 ligands (Table\u00a02II cation lie in the range 82.63\u2005(14) to 175.09\u2005(10)\u00b0. Intra\u00admolecular N2\u2014H2B\u22efN3 and N4\u2014H4B\u22efN3 hydrogen bonds are also present (Table\u00a04When MnSOs Table\u00a02. In contt Table\u00a04.2L1) acts as a monodentate ligand. The results indicate that the rational design and selection of ligands with heterocyclic nitro\u00adgen systems is an effective synthetic strategy to construct complexes via self-assembly. The asymmetry of the [N,N\u2032-bis\u00ad(pyridin-4-yl)pyridine-2,6-dicarboxamide ligand has resulted in some novel structures. Further study is ongoing.In complexes (I)2L1 ligands play a crucial role in mol\u00adecular self-assembly, with centroid-to-centroid separations of 3.5808\u2005(13) and 3.6269\u2005(14)\u2005\u00c5. In addition, a number of O\u2014H\u22efN and O\u2014H\u22efO hydrogen-bonding inter\u00adactions (Table\u00a03In (I)s Table\u00a03 connect s Table\u00a03.via O\u2014H\u22efN and O\u2014H\u22efO hydrogen-bonding inter\u00adactions (Table\u00a042L1 ligand are important in molecular self-assembly.Complex (II)s Table\u00a04. In (II)et al., 20162L1 skeleton. These include the methyl pyridinium compound of H2L1 was prepared using a modified literature procedure in N,N\u2032-di\u00admethyl\u00adformamide solution (4\u2005mL) was gradually added to MnSO4.H2O in a mixed solution . After standing for 5\u2005min, the suspension was filtered and the filtrate was kept at room temperature in the dark. One week later, colourless single crystals suitable for X-ray diffraction were obtained. Complex (II)4.8/3H2O was used instead of MnSO4.H2O.The heterocyclic nitro\u00adgen ligand (HUiso(H) = 1.2\u20131.5Ueq(C/N/O).Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989018007351/cq2024sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S2056989018007351/cq2024Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989018007351/cq2024IIsup3.hklStructure factors: contains datablock(s) II. DOI: 1822492, 1822490CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Two chalcones were synthesized in Claisen\u2013Schmidt condensation reactions. In the crystals, \u03c0\u2013\u03c0 inter\u00adactions and weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions are observed. The effect of these inter\u00admolecular inter\u00adactions in the solid state can be seen inthe difference between the experimental and theoretical optimized geometrical parameters. 31H23NO and C35H23NO, were synthesized via Claisen\u2013Schmidt condensation reactions. Both structures were solved and refined using single-crystal X-ray diffraction data and optimized at the ground state using the density functional theory (DFT) method with the B3LYP/6-311++G level. In the crystals, \u03c0\u2013\u03c0 inter\u00adations and weak C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions are observed. The effect of these inter\u00admolecular inter\u00adactions in the solid state can be seen by the difference between the experimental and theoretical optimized geometrical parameters. The structures have also been characterized by UV\u2013Vis spectroscopy. The smallest energy gaps of 2.86 and 2.96\u2005eV enhance the nonlinear responses of such mol\u00adecular systems. Hirshfeld surface analyses and 2D fingerprint plots were used to qu\u00adantify the inter\u00admolecular inter\u00adactions present in the crystal, indicating that these are the most important contribution to the crystal packing.The title chalcones, C Additionally, the UV\u2013vis absorption and Hirshfeld surface analyses are discussed.Chalcones satisfy the criteria of three features essential for high nonlinear activity in an organic compound, which are: a strong electron donor, a highly polarizable \u03c0-conjugated bridge and a strong \u03c0-electron acceptor. A chalcone mol\u00adecule with a \u03c0-conjugated system provides a large charge-transfer axis with appropriate substituent groups on the terminal aromatic rings. Polyaromatic hydro\u00adcarbons or \u03c0-conjugated materials such as anthracenyl chalcone provide the significant property for conductivity that led to tremendous advances in the field of organic electronics . The C\u2014C distances in the central ring of the anthracene units show a little variations compared to the other rings , which are much shorter. These observations are consistent with an electronic structure for the anthracene units where a central ring displaying aromatic delocalization is flanked by two isolated diene units \u2005\u00c5 and DFT = 1.22\u2005\u00c5 in I; Exp = 1.213\u2005(3) (A) and 1.218\u2005(3)\u2005\u00c5 (B), and DFT\u00a0= 1.22\u2005\u00c5 in II] and C16=C17 . Both I and II (A and B) are twisted at the C14\u2014C15 bond, with C1\u2014C14\u2014C15\u2014C16 torsion angles of \u221292.6\u2005(2) (in I), 84.8\u2005(3) (in IIA) and 106.3\u2005(3)\u00b0 (in IIB). The corresponding torsion angles for DFT are \u221285.84 and 85.63\u00b0, respectively. Additionally, in compound II, rings Y and Z (A) and rings Y\u2032 and Z\u2032 (B) are also twisted at the C21\u2014N1 bond, with C20\u2014C21\u2014N1\u2014C24 torsion angles of Exp = 64.1\u2005(4)\u00b0 (A) and 46.2\u2005(4)\u00b0 (B), and DFT = 55.03\u00b0. The large twist angles are due to the bulkiness of the strong electron-donor anthracene ring system and substituent ring system \u00b0 and DFT\u00a0= \u22121.38 for compound I, and Exp = \u2212171.2\u2005(3)\u00b0 (A) and 11.4\u2005(5)\u00b0 (B), and DFT = \u22121.70\u00b0 for compound II. The slight differences in the torsion angles between the experimental and DFT results in both compounds are due to the formation of inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions involving all the fused-ring systems, which are not taken into consideration during the optimization process \u2005\u00c5 at atom C16] makes dihedral angles of 86.93\u2005(19) and 21.21\u2005(19)\u00b0 with the anthracene ring [maximum deviation = 0.0117\u2005(19)\u2005\u00c5 at C9] and ring X [maximum deviation = 0.0363\u2005(18)\u2005\u00c5 at C29], respectively. In compound II, the enone moiety for mol\u00adecule A forms dihedral angles of 84.76\u2005(17), 87.61\u2005(17) and 72.35\u2005(17)\u00b0 with the anthracene ring [maximum deviation = 0.029\u2005(3)\u2005\u00c5 at C14A], ring Y [maximum deviation = 0.008\u2005(3)\u2005\u00c5 at C19A] and ring Z [maximum deviation = 0.043\u2005(3)\u2005\u00c5 at C34A], respectively. The anthracene ring forms dihedral angles of 89.63\u2005(11) and 62.11\u2005(7)\u00b0 with rings Y and Z, respectively, and the dihedral angle between rings Y and Z is 61.73\u2005(10)\u00b0. In addition, for mol\u00adecule B, the enone moiety forms dihedral angles of 72.2\u2005(3), 13.5\u2005(3) and 87.2\u2005(3)\u00b0 with the anthracene ring [maximum deviation = 0.018\u2005(4)\u2005\u00c5 at C10B], ring Y\u2032 [maximum deviation = 0.010\u2005(3)\u2005\u00c5 at C20B] and ring Z\u2032 [maximum deviation = 1.441\u2005(2)\u2005\u00c5 at N1B], respectively. The anthracene ring forms dihedral angles of 61.46\u2005(11) and 54.80\u2005(7)\u00b0 with rings Y\u2032 and Z\u2032, respectively, and the dihedral angle between rings Y\u2032 and Z\u2032 is 48.92\u2005(11)\u00b0.The enone moiety in I shows weak \u03c0\u2013\u03c0 inter\u00adactions involving Cg1\u22efCg5 = 3.7267\u2005(11)\u2005\u00c5 , Cg2\u22efCg4 = 3.6669\u2005(12)\u2005\u00c5 , Cg3\u22efCg3 = 3.6585\u2005(11)\u2005\u00c5 and Cg4\u22efCg4 = 3.6790\u2005(12)\u2005\u00c5 , where Cg1, Cg2, Cg3, Cg4 and Cg5 are the centroids of rings N1/C20/C21/C26/C27, C1\u2013C6, C1/C6\u2013C8/C13/C14, C8\u2013C13, C18\u2013C20/C27-C29, respectively. The packing is further linked into an infinite three-dimensional supra\u00admolecular network.The crystal packing of ns Fig.\u00a02a involvII shows weak C12B\u2014H12B\u22efO1 inter\u00admolecular hydrogen bonds connecting the mol\u00adecules into an infinite one-dimensional chain along the c axis. In addition, weak inter\u00admolecular C5B\u2014H5BA\u22efCg6, C27B\u2014H27B\u22efCg7, C28B\u2014H28B\u22efCg8, C11A\u2014H11A\u22efCg9 and C7B\u2014H7B\u22efCg10 inter\u00adactions are also observed in the crystal packing and further stabilize the crystal structure, where Cg6, Cg7, Cg8, Cg9 and Cg10 are the centroids of rings N1A/C24A/C29A/C30A/C35A, C1A\u2013C6A, C1A/C6A\u2013C8A/C13A/C14A, C18A\u2013C23A and C24A\u2013C29A, respectively. These weak inter\u00admolecular C\u2014H\u22efO and C\u2014H\u22ef\u03c0 inter\u00adactions bridge the mol\u00adecules into an infinite one-dimensional column along the c axis.Lists of weak hydrogen-bond inter\u00admolecular inter\u00adactions are shown in Table\u00a01II Fig.\u00a01b shows I and II have been calculated using time-dependent DFT at the B3LYP/6-311++G level in the gas phase and give values of 396 (I) and 383\u2005nm (II). The absorption characteristics of I and II are observed in the UV region at 393 and 388\u2005nm, as shown in Fig.\u00a03et al., 2017The electronic absorption spectra of I and II, respectively. Through an extrapolation of the linear trend observed in the optical spectra, the experimental energy band gaps in I and II are 2.86 and 2.96\u2005eV, respectively. These optical band-gap values indicate the suitability of this compound for optoelectronic applications, as was also reported previously for a chalcone structure by Tejkiran et al. energy levels of the title compounds, the corresponding electronic transfer are found to happen between the HOMO and LUMO orbitals, as shown in Fig.\u00a04 al. 2016. In addi al. 2016 studied CrystalExplorer show the HS mapped over dnorm, where the red spots indicate the regions of donor\u2013acceptor inter\u00adactions. The C\u2014H\u22efO contacts are only present in compound II. In addition, the presence of C\u2014H\u22ef\u03c0 inter\u00adactions only occurs in compound II, indicated through a combination of pale-orange bright-red spots which are present on the HS mapped over shape index surface, identified with black arrows . The large flat region delineated by a blue outline refers to the \u03c0\u2013\u03c0 stacking inter\u00adactions. The curved nature of the compound reveals that \u03c0\u2013\u03c0 stacking inter\u00adactions are present in compound I. Meanwhile, these inter\u00adactions are absent in compound II.The program ws Fig.\u00a05b. The lII. Meanwhile, there is no spike in the fingerprint of compound I. The 7.5% O\u22efH contribution shown in compound I is the average percentage interaction from the total interactions presence in I. In compound I, there are no interactions other than the \u03c0\u2013\u03c0 interactions, which makes the percentage of the O\u22efH contribution is slightly higher. Hence, a discussion on the percentage difference between I. and II. is invalid. The significant C\u2014H\u22ef\u03c0 inter\u00adactions for compound II are indicated by the wings de + di \u223c 2.6\u2005\u00c5.The fingerprint plot shown in Fig.\u00a06et al., 2016I and II. There are four compounds which have an anthrancene ketone subtituent on the chalcone, including 9-anthryl styryl ketone and 9,10-anthryl bis\u00ad(styryl ketone) reported by Harlow et al. (1975E)-1-(Anthracen-9-yl)-3-[4-(propan-2-yl)phen\u00adyl]prop-2-en-1-one was reported by Girisha et al. (2016E)-1-(anthracen-9-yl)-3-(2-chloro-6-fluoro\u00adphen\u00adyl)prop-2-en-1-one was reported by Abdullah et al. -1,3-bis\u00ad(anthracen-9-yl)prop-2-en-1-one. Other related compounds include 1-(anthracen-9-yl)-2-methyl\u00adprop-2-en-1-one -1-(anthracen-9-yl)-3-[4-(piperidin-1-yl)phen\u00adyl]prop-2-en-1-one and (E)-1-(anthracen-9-yl)-3-[4-(di\u00adphenyl\u00adamino)\u00adphen\u00adyl]prop-2-en-1-one -1- al. 2016, while benzaldehyde (0.5\u2005mmol) for compounds I and II, respectively, was dissolved in methanol (20\u2005ml). A catalytic amount of NaOH was added to the solution dropwise under vigorous stirring. The reaction mixture was stirred for about 5\u20136\u2005h at room temperature. After stirring, the contents of the flask were poured into ice-cold water (50\u2005ml). The resultant crude products were filtered, washed successively with distilled water and recrystallized from acetone to give the corresponding chalcones (Scheme 1). Single crystals of I and II suitable for X-ray diffraction were obtained by slow evaporation from acetone solutions.A mixture of 9-acetyl\u00adanthrancene (0.5\u2005mmol) and 9-ethylcarbazole-3-carbaldehyde (0.5\u2005mmol) and 4- and refined using a riding model, with Uiso(H) = 1.2 or 1.5Ueq(C). A rotating group model was applied to the methyl group in I.Crystal data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018011131/lh5878sup1.cifCrystal structure: contains datablock(s) mo_DA20_0m, mo_DA21e_0m, global. DOI: 10.1107/S2056989018011131/lh5878mo_DA20_0msup2.hklStructure factors: contains datablock(s) mo_DA20_0m. DOI: Click here for additional data file.10.1107/S2056989018011131/lh5878mo_DA20_0msup4.cmlSupporting information file. DOI: 10.1107/S2056989018011131/lh5878mo_DA21e_0msup3.hklStructure factors: contains datablock(s) mo_DA21e_0m. DOI: 10.1107/S2056989018011131/lh5878sup5.pdfSupporting information file. DOI: 1827021, 1827019CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The crystal structures of two multidentate CMPO-containing organic ligands are described. Both compounds feature N\u2014H\u22efO hydrogen bonds in the solid state. 36H34N2O4P2, (I), and diethyl [({2-[2-(di\u00adeth\u00adoxy\u00adphosphino\u00adyl)ethanamido]\u00adeth\u00adyl}carbamo\u00adyl)meth\u00adyl]phospho\u00adnate, C14H30N2O8P2, (II), were synthesized via nucleophilic acyl substitution reactions between an ester and a primary amine. Hydrogen-bonding inter\u00adactions are present in both crystals, but these inter\u00adactions are intra\u00admolecular in the case of compound (I) and inter\u00admolecular in compound (II). Intra\u00admolecular \u03c0\u2013\u03c0 stacking inter\u00adactions are also present in the crystal of compound (I) with a centroid\u2013centroid distance of 3.9479\u2005(12)\u2005\u00c5 and a dihedral angle of 9.56\u2005(12)\u00b0. Inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions give rise to supra\u00admolecular sheets that lie in the ab plane. Key geometric features for compound (I) involve a nearly planar, trans-amide group with a C\u2014N\u2014C\u2014C torsion angle of 169.12\u2005(17)\u00b0, and a torsion angle of \u2212108.39\u2005(15)\u00b0 between the phosphine oxide phospho\u00adrus atom and the amide nitro\u00adgen atom. For compound (II), the electron density corresponding to the phosphoryl group was disordered, and was modeled as two parts with a 0.7387\u2005(19):0.2613\u2005(19) occupancy ratio. Compound (II) also boasts a trans-amide group that approaches planarity with a C\u2014N\u2014C\u2014C torsion angle of \u2212176.50\u2005(16)\u00b0. The hydrogen bonds in this structure are inter\u00admolecular, with a D\u22efA distance of 2.883\u2005(2)\u2005\u00c5 and a D\u2014H\u22efA angle of 175.0\u2005(18)\u00b0 between the amide hydrogen atom and the P=O oxygen atom. These non-covalent inter\u00adactions create ribbons that run along the b-axis direction.Two bis-carbamoyl\u00admethyl\u00adphosphine oxide compounds, namely {[(3-{[2-(di\u00adphen\u00adyl\u00adphosphino\u00adyl)ethanamido]\u00admeth\u00adyl}benz\u00adyl)carbamo\u00adyl]meth\u00adyl}di\u00adphenyl\u00adphos\u00adphine oxide, C Many researchers have attempted to mimic this solution stoichiometry by tethering two, three or four CMPO groups together via an organic scaffold 1H, 13C, and 31P NMR spectroscopy, and by X-ray crystallography.The carbamoyl\u00admethyl\u00adphosphine oxide (CMPO) moiety has found use as the chelating portion of a ligand in the TRUEX process for the remediation of nuclear waste 4 descriptor for fourfold coordination around the phospho\u00adrus atom P1 is 0.95, indicating a nearly perfect tetra\u00adhedral geometry of the phosphine oxide group \u00b0, and is staggered with respect to the plane of the C4\u2013C7 aromatic ring with a H1\u2014N1\u2014C3\u2014C4 torsion angle of 59.1\u2005(17)\u00b0.The structure of compound (I)x\u00a0+\u00a01, y, \u2212z\u00a0+\u00a0D\u22efA distance of 2.940\u2005(2)\u2005\u00c5 and a D\u2014H\u22efA angle of 168\u2005(2)\u00b0 )\u00b0 Fig.\u00a03. The C14Pbca. Since the mol\u00adecule lies on an inversion center , the asymmetric unit comprises one half of the mol\u00adecule. The electron density corresponding to the atoms of the phosphoryl group was disordered and was modeled over two positions with a 0.7387\u2005(19):0.2613\u2005(19) occupancy ratio (see the Refinement section for more details). The complete mol\u00adecular structure of the major component of compound (II)4 descriptor for fourfold coordination around the phospho\u00adrus atom of the major component, P1, is 0.93, indicating that the geometry of the phosphoryl group is slightly distorted from an ideal tetra\u00adhedron. The geometry between the amide nitro\u00adgen atom N1 and the \u03b2-phosphoryl group phospho\u00adrus atom P1 is defined by a N1\u2014C1\u2014C2\u2014P1 torsion angle of \u2212111.8\u2005(2)\u00b0. The amide group of this compound also adopts a nearly perfect trans geometry with a C3\u2014N1\u2014C1\u2014C2 torsion angle of \u2212176.50\u2005(16)\u00b0.Compound (II)A with a C\u22efcentroid distance of 3.622\u2005(2)\u2005\u00c5 and a C\u2014H\u22efcentroid angle of 146\u00b0. These non-covalent inter\u00adactions create supra\u00admolecular sheets of compound (I)ab plane ne Fig.\u00a04.x\u00a0+\u00a0y\u00a0+\u00a0z\u00a0+\u00a01; Fig.\u00a05D\u22efA distance of 2.883\u2005(2)\u2005\u00c5 with a D\u2014H\u22efA angle of 175.0\u2005(18)\u00b0. This hydrogen bond forms ribbons of compound (I)b-axis direction on Fig.\u00a06.et al., 2016via an organic scaffold. The most similar structures to compound (I)et al., 2014et al., 2006via inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions. A structure closely related to compound (II)et al., 20023)3 has been reported in this journal 1,3-Bis(amino\u00admeth\u00adyl)benzene and the p-nitro\u00adphenyl ester of di\u00adphenyl\u00adphosphono\u00adacetate and the solution was stirred for 3.5\u2005h. The organic layer was separated, washed with brine (3 \u00d7 10\u2005mL), dried over solid magnesium sulfate and concentrated under reduced pressure. The crude product was triturated multiple times with ethyl acetate to give a white solid in 91% yield. X-ray quality crystals of compound (I)1H NMR : \u03b4 7.91 , 7.7\u20137.3 , 7.1\u20136.8 , 4.24 , 3.36 ; 13C NMR : \u03b4 164.7 , 138.3, 132.5, 131.9, 131.2\u2013130.5 (broad), 129.5\u2013128.3 (broad), 126.9\u2013126.1 (broad), 43.5, 38.6; 31P NMR : \u03b4 30.6.Compound (II) Ethyl\u00adene di\u00adamine was dissolved in 8.3\u2005mL of methanol. The solution was cooled to 195\u2005K, and triethyl phosphono\u00adacetate was added dropwise. The reaction mixture was allowed to warm to room temperature and stirred overnight. The product precipitated from the solution, was isolated by vacuum filtration and rinsed with ethyl acetate. Some of this solid was crystalline and suitable for analysis by X-ray diffraction. The remainder of the isolated product was purified by silica gel chromatography (10:1 di\u00adchloro\u00admethane\u2013methanol) to give compound (II)1H NMR : \u03b4 7.75 , 4.15 , 3.34 , 2.85 , 1.33 ; 13C NMR : \u03b4 165.4, 62.9, 35.8 , 16.5; 31P NMR : \u03b4 24.5.Uiso(H) = 1.2Ueq(C) for methyl\u00adene groups and aromatic hydrogen atoms, and Uiso(H) = 1.5Ueq(C) for methyl groups. For both compounds (I)SHELXL were treated with SAME and EADP commands to produce bond lengths and angles that agree with known values, and to ensure physically reasonable displacement parameters.Crystal data, data collection and structure refinement details for both compounds are summarized in Table\u00a0310.1107/S205698901900820X/pk2617sup1.cifCrystal structure: contains datablock(s) I, II. DOI: 10.1107/S205698901900820X/pk2617Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S205698901900820X/pk2617Isup4.cmlSupporting information file. DOI: 10.1107/S205698901900820X/pk2617IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S205698901900820X/pk2617IIsup5.cmlSupporting information file. DOI: 1921486, 1921485CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "In the crystal structure of the title compound, two half-mol\u00adecules are found in the asymmetric unit. The completed mol\u00adecules differ only slightly in bond lengths and torsion angles. 14H14N2S2, was obtained by transmetallation of 2,2\u2032-bis\u00ad(tri\u00admethyl\u00adstann\u00adyl)azo\u00adbenzene with methyl lithium, and subsequent quenching with dimethyl di\u00adsulfide. The asymmetric unit comprises two half-mol\u00adecules, the other halves being completed by inversion symmetry at the midpoint of the azo group. The two mol\u00adecules show only slight differences with respect to N=N, S\u2014N and aromatic C=C bonds or angles. Hirshfeld surface analysis reveals that except for one weak H\u22efS inter\u00adaction, inter\u00admolecular inter\u00adactions are dominated by van der Waals forces only.The title compound, C Azo\u00adbenzenes are common motifs in dyes because of their high thermal and photochemical stability 2, (I)The mol\u00adecular switch azo\u00adbenzene can undergo isomerization from its thermodynamically stable Ia and Ib), the other halves being completed by application of inversion symmetry. The midpoints of the N=N bonds are located on inversion centres, resulting in a trans-configuration for the central N=N bonds \u2212x, 1\u00a0\u2212\u00a0y, \u2212z; (ii) 1\u00a0\u2212\u00a0x, 1\u00a0\u2212\u00a0y, 1\u00a0\u2212\u00a0z] torsion angles of 13.2\u2005(2) and \u22125.3\u2005(2)\u00b0, respectively, in both mol\u00adecules the phenyl rings are twisted slightly with respect to the azo unit. A weak distortion is also found for the N1\u2014C1\u2014C2\u2014S1 torsion angles of \u22123.06\u2005(16)\u00b0 for Ia and \u22122.06\u2005(15)\u00b0 for Ib. The N=N bond lengths differ marginally , as do comparable C\u2014C bonds. For example, the C1\u2014C2 bond in Ia is at 1.408\u2005(2)\u2005\u00c5 slightly shorter than Ib [1.415\u2005(2)\u2005\u00c5]. In comparison, this bond is longer than all other C\u2014C distances in the ring because of repulsion of the nitro\u00adgen and the sulfur atoms attached to C1 and C2, respectively. In both mol\u00adecules, the S\u22efN distances are too long to be considered as attractive inter\u00adactions. Fig.\u00a02The asymmetric unit of the title compound consists of two half-mol\u00adecules = 3.7525\u2005(8)\u2005\u00c5 with a slippage of 1.422\u2005\u00c5. To further investigate the inter\u00admolecular inter\u00adactions, Hirshfeld surfaces of another mol\u00adecule of Ib (S\u22efH distance = 2.811\u2005\u00c5). The two-dimensional fingerprint plots for mol\u00adecule Ib for qu\u00adanti\u00adfication of the contributions of each type of non-covalent inter\u00adaction to the Hirshfeld surface azo\u00adbenzene was recently described was purchased from Acros Organics. THF was purchased from VWR and was dried and degassed with a solvent purification system by Inert Technology.The synthesis of 2,2\u2032- al. 2015. Dimethy2,2\u2032-bis\u00ad(Methyl\u00adthio)\u00adazo\u00adbenzenebis(tri\u00admethyl\u00adstann\u00adyl)azo\u00adbenzene was dissolved under Schlenk conditions in THF (12.5\u2005ml) and cooled to 195\u2005K. Then MeLi was added within 5\u2005min and after 1.5\u2005h at this temperature, dimethyl di\u00adsulfide was added in one ration. The reaction mixture was warmed to 298\u2005K over 14\u2005h and the solvent was removed under reduced pressure. The obtained orange solid was purified in a silica column with a gradient of eluents from n-pentane to di\u00adchloro\u00admethane giving dark-orange crystals . Single crystals suitable for X-ray analysis were obtained by slow evaporation from a saturated n-heptane solution.In an inert reaction tube, 2,2-1H NMR : \u03b4 = 7.76 , 7.40 , 7.32 , 7.20 , 2.53 ppm.13C{1H} NMR : \u03b4 = 149.08 (C1), 141.00 (C2), 131.56 (C4), 124.81 (C3), 124.75 (C5), 118.02 (C6), 15.02 (C7) ppm.HRMS : m/z: calculated for C14H14N2S2+ 274.05929 found 274.05944.MS (EI): m/z 273.9 (5%) [M]+, 258.9 (100%) [M\u00a0\u2212\u00a0CH3]+, 243.9 (5%) [M\u00a0\u2212\u00a0C2H6]+, 107.9 (13%) [M\u00a0\u2212\u00a0C8H10N2S]+.IR (ATR): \u03bd = 3059 (w), 2986 (w), 2961 (w), 2918 (w), 2852 (w), 1575 (m), 1561 (w), 1457 (m), 1433 (s), 1298 (w), 1249 (w), 1217 (m), 1162 (m), 1065 (s), 1035 (m), 951 (m), 863 (w), 803 (w), 761 (s), 726 (s), 674 (s) cm\u22121.M.p.: 429\u2005KRf: (n-penta\u00adne: di\u00adchloro\u00admethane 3:1): 0.55.Uiso(H) = 1.5Ueq (C-meth\u00adyl) and 1.2Ueq(C) (C-phen\u00adyl).Crystal data, data collection and structure refinement details are summarized in Table\u00a0110.1107/S2056989019014592/wm5522sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019014592/wm5522Isup2.hklStructure factors: contains datablock(s) I. DOI: 1961741, 1961741CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "N\u2032,N\u2032\u2032\u2032--{[methyl\u00adenebis(\u00adoxy)]bis\u00ad}bis\u00ad(methane\u00adylyl\u00adidene))bis\u00ad(isonicotinohydrazide) dihydrate, (I), and N\u2032,N\u2032\u2032\u2032--{bis\u00ad}bis\u00ad(methane\u00adylyl\u00adidene))bis\u00ad(isonicotinohydrazide), (II), both crystallized with half a mol\u00adecule in the asymmetric unit. The whole mol\u00adecule of (I) is generated by twofold rotation symmetry, with the twofold rotation axis bis\u00adecting the C atom of the \u2013O\u2014CH2\u2014O\u2013 bridge. The whole mol\u00adecule of (II) is generated by inversion symmetry, with the central CH2\u2014CH2 bond of the \u2013O\u2014(CH2)4\u2014O\u2013 bridge being located about a center of inversion.The title compounds, 27H20Br2N6O4\u00b72H2O, (I), and C30H28N6O4\u00b7[+ solvent], (II), both crystallize with one half-mol\u00adecule in the asymmetric unit. The whole mol\u00adecule of (I) is generated by twofold rotation symmetry, with the twofold rotation axis bis\u00adecting the C atom of the \u2013O\u2014CH2\u2014O\u2013 bridge. This results in a folded or U-shaped conformation of the mol\u00adecule. The whole mol\u00adecule of (II) is generated by inversion symmetry, with the central CH2\u2014CH2 bond of the \u2013O\u2014(CH2)4\u2014O\u2013 bridge being located about a center of inversion. This results in a step-like conformation of the mol\u00adecule. The central C(=O)N\u2014N=C regions of the isonicotinohydrazide moieties in both compounds are planar and the configuration about the imine C=N bonds is E. In compound (I), the benzene and pyridine rings are inclined to each other by 37.60\u2005(6)\u00b0. The two symmetry-related pyridine rings are inclined to each other by 74.24\u2005(6)\u00b0, and the two symmetry-related benzene rings by 7.69\u2005(6)\u00b0. In compound (II), the benzene and pyridine rings are inclined to each other by 25.56\u2005(11)\u00b0. The symmetry-related pyridine rings are parallel, as are the two symmetry-related benzene rings. In the crystal of (I), a pair of water mol\u00adecules link the organic mol\u00adecules via Owater\u2014H\u22efO and Owater\u2014H\u22efN hydrogen bonds, forming chains along [001], and enclosing an R42(8) and two R12(5) ring motifs. The chains are linked by N\u2014H\u22efNpyridine hydrogen bonds, forming a supra\u00admolecular framework. There are also a number of C\u2014H\u22efO hydrogen bonds, and C\u2014H\u22ef\u03c0 and offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adplanar distance = 3.294\u2005(1)\u2005\u00c5] present reinforcing the framework. In the crystal of (II), mol\u00adecules are linked by N\u2014H\u22efNpyridine hydrogen bonds, forming a supra\u00admolecular framework. Here too there are also a number of C\u2014H\u22efO hydrogen bonds present, and a C\u2014H\u22ef\u03c0 inter\u00adaction, reinforcing the framework. For compound (II), a region of disordered electron density was corrected for using the SQUEEZE [Spek (2015PLATON. Their formula mass and unit-cell characteristics were not taken into account during refinement.The title compounds, CSpek 2015. Acta Cr The C6\u2014N2 and C7=N3 bond lengths differ by 0.068\u2005(2)\u2005\u00c5 in (I)et al., 2013et al., 2013et al., 2010The bond lengths and angles in the carbohydrazide group of the title compounds can be compared with the values reported for related structures \u2005\u00c5] present, reinforcing the framework is 3.766\u2005(1)\u2005\u00c5, \u03b1 = 0.00\u2005(6)\u00b0, \u03b2 = 29\u00b0, inter\u00adplanar distance = 3.294\u2005(1)\u2005\u00c5, offset of 1.824\u2005\u00c5.In the crystal of (I)s Table\u00a03. The chak Table\u00a03. The offpyridine hydrogen bonds, forming a supra\u00admolecular framework k Table\u00a04. Here tok Table\u00a04, reinfor3 with an electron count of 357 per unit cell was corrected for using the SQUEEZE routine in PLATON et al., 2007CrystalExplorer17 through white to blue . The dnorm surface was mapped over a fixed colour scale of \u22120.512 (red) to 1.285 (blue) for compound (I)SQUEEZED out).The Hirshfeld surfaces of compounds (I)b), O\u22efH/H\u22efO at 13.8% , N\u22efH/H\u22efN at 11.3% , Br\u22efH/H\u22efBr at 14.3% and C\u22efH/H\u22efC contacts at 13.6% . C\u22efC contacts account for 8.4%, while C\u22efBr are 3.0%, C\u22efN are 3.0%, and finally C\u22efO contacts amount to 1.4%.The fingerprint plots are given in Figs. 98% Fig.\u00a09c, N\u22efH/H3% Fig.\u00a09d, Br\u22efH/3% Fig.\u00a09e and C\u22ef6% Fig.\u00a09f. C\u22efC cb), O\u22efH/H\u22efO at 13.3% , N\u22efH/H\u22efN at 16.2% , and C\u22efH/H\u22efC at 33.6% . The remaining contacts are extremely weak, ca 1% each.For compound (II)% Fig.\u00a010c, N\u22efH/H% Fig.\u00a010d, and C% Fig.\u00a010e. The ret al., 2016i.e. (3-OR-benzyl\u00adidene)isonicotinohydrazide (R = C) skeleton gave 51 hits (supporting information file S1). The majority of these compounds were with an OMe or an OEt substituent.A search of the Cambridge Structural Database isonicotinohydrazide (R = C) skeleton gave 23 hits (supporting information file S2). Again, the majority of these compounds have an OMe or an OEt substituent. However, here the most inter\u00adesting and relevant compound concerns the ligand N\u2032,N\u2032\u2032-bis\u00ad(pyridine-4-carbohydrazide), in which a 1,2-di\u00adoxy\u00adethane bridge links two N\u2032-benzyl\u00adideneisonicotinohydrazide units. The crystal structures of two polymorphs have been described: a monoclinic P21 polymorph that crystallizes as a methanol disolvate ] bis(pyridine-4-carbohydrazide)})tetra\u00adkis\u00ad(iodo)\u00addimercury meth\u00adanol disolvate , in a 250\u2005ml round-bottom (RB) flask was added DMF (30\u2005ml) and potassium carbonate (12.5\u2005mmol). The mixture was stirred at room temperature and then 1,1-di\u00adiodo\u00adbutane (2.5\u2005mmol) was added dropwise and the reaction mixture was stirred for 12\u2005h. It was then partitioned between water and ethyl acetate. The ethyl acetate layer was collected and concentrated under reduced pressure. To 1,4-bis\u00adbutane (2\u2005mmol) and isonicotinic acid hydrazide (4\u2005mmol) in a 250\u2005ml RB flask was added 100\u2005ml of methanol and two drops of glacial acetic acid. The reaction mixture was stirred at room temperature and within 5\u2005min a white-coloured product had formed. The reaction was continued for a further 30\u2005min. The title compound was isolated by filtration and washed with methanol, then chloro\u00adform and followed by acetone. The final product was recrystallized using DMSO and yielded colourless block-like crystals of compound (I)Compound I: To 5-bromo-2-hy\u00addroxy\u00adbenzaldehyde (5\u2005mmol), in a 250\u2005ml RB flask, was added 50\u2005ml of DMF and potassium carbonate (12.5\u2005mmol). The mixture was stirred at room temperature and then 1,1-di\u00adiodo\u00admethane (2.5\u2005mmol) was added dropwise. Then, the reaction mixture was stirred for 12\u2005h. The product obtained was extracted in ethyl acetate medium. Methanol (100\u2005ml) and two drops of glacial acetic acid were added to a mixture of 6,6\u2032-[methyl\u00adenebis(\u00adoxy)] bis\u00ad (2\u2005mmol) and isoniazid (4\u2005mmol) in a 250\u2005ml RB flask. The reaction mixture was stirred at room temperature and within 5\u2005min a white-coloured product had formed and the reaction was continued for a further 30\u2005min. The solid obtained was washed with methanol, then chloro\u00adform and followed by acetone. The final product was recrystallized using DMSO and yielded colourless block-like crystals of compound (II)Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a053 with an electron count of 357 per unit cell was corrected for using the SQUEEZE routine in PLATON 10.1107/S2056989019005048/su5495sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989019005048/su5495Isup4.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019005048/su5495IIsup5.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989019005048/su5495Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019005048/su5495IIsup5.cmlSupporting information file. DOI: 10.1107/S2056989019005048/su5495sup6.pdfCSD search S1. DOI: 10.1107/S2056989019005048/su5495sup7.pdfCSD Search S2. DOI: 1907912, 1907913CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III atoms in the title structure have distorted octa\u00adhedral coordination spheres, being C,N-chelated by two main 2-phenyl\u00adpyridine ligands and N,O-chelated by one ancillary 2-[\u00admeth\u00adyl]phenolato ligand.The two Ir 11H8N)2(C15H14NO3)]\u00b72CH2Cl2, consists of two complex mol\u00adecules together with four di\u00adchloro\u00admethane solvent mol\u00adecules, one of which is disordered. In each complex mol\u00adecule, the IrIII ion has a distorted octa\u00adhedral coordination environment defined by two 2-phenyl\u00adpyridine ligands, through two phenyl C and two pyridine N atoms, and by one N,O-bidentate 2-[\u00admeth\u00adyl]phenolate anion. The IrIII ions lie almost in the equatorial planes with deviations of 0.0396\u2005(17) and 0.0237\u2005(17)\u2005\u00c5, respectively, for the two complex mol\u00adecules. In both complex mol\u00adecules, the two 2-phenyl\u00adpyridine ligands are nearly perpendicular to each other [dihedral angles between the least-squares-planes of 89.91\u2005(11) and 85.13\u2005(11)\u00b0]. In the crystal, inter\u00admolecular C\u2014H\u22efO inter\u00adactions as well as inter\u00admolecular C\u2014H\u22ef\u03c0 inter\u00adactions are present, leading to a three-dimensional network structure. One of the four dichlormethane solvent mol\u00adecules shows disorder over two sets of sites [occupancy ratio 0.79\u2005(2):0.21\u2005(2)].The asymmetric unit of the solvated title complex, [Ir(C The equatorial plane is formed by atoms O4/C49/C38/N6. The mean deviation from the least-squares plane is 0.055\u2005\u00c5 and the Ir2III ion is displaced by 0.0237\u2005(17)\u2005\u00c5 from the equatorial plane towards the axial N4 atom. The deviation from a perpendicular arrangement of the two 2-phenyl\u00adpyridine ligands is slightly higher than in complex 1 [the dihedral angle between the least-squares planes is 85.13\u2005(11)\u00b0], likewise the deviation from planarity with dihedral angles of 1.69\u2005(13)\u00b0 (between rings C49\u2013C54 and N5\u2013C59) and 3.36\u2005(13)\u00b0 (between rings C38\u2013C43 and N4\u2013C48), respectively.In complex mol\u00adecule 2 (Ir2), a similar bonding situation is observed, with the phenyl C atoms C38 and C49 Cg1 and H50 with Cg2 .The configurations in both complexes are stabilised by intra\u00admolecular C\u2014H\u22efO inter\u00adactions between the phenolic O1 and O4 atoms as acceptors and the phenyl C1\u2014H1 and C48\u2014H48 groups as donors Fig.\u00a01, as welle.g. for {(E)-2-[\u00admeth\u00adyl]phen\u00ado\u00adlato-\u03ba2N,O}bis\u00adiridium(III) -2-[(phenyl\u00adimino)\u00admeth\u00adyl]phenolato-\u03ba2N,O}bisiridium(III) -2-[(phenyl\u00adimino)\u00admeth\u00adyl]phenolato-\u03ba2N,O}bis\u00adiridium(III) , although the title compound is very similar to a previously reported compound into a di\u00adchloro\u00admethane (5\u2005mL) solution of the title compound at room temperature.The title compound was prepared according to a reported procedure (Goo sp2\u2014H and 0.99\u2005\u00c5 for methyl\u00adene C\u2014H with Uiso(H) = 1.2Ueq(C); C\u2014H = 0.98\u2005\u00c5 with Uiso(H) = 1.5Ueq(C) for methyl H atoms. One of the four dichloro\u00admethane solvent mol\u00adecules shows disorder over two sets of sites [occupancy ratio 0.79\u2005(2):0.21\u2005(2)].Crystal data, data collection and structure refinement details are summarized in Table\u00a0310.1107/S2056989018009970/wm5449sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018009970/wm5449Isup2.hklStructure factors: contains datablock(s) I. DOI: 1846724CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "III{C(dppm)2-\u03ba3P,C,P\u2032}ClH(NH3C2)]Cl with ethyl diazo\u00adacetate, a well known C=C coupling reagent, leads to the formation of a C=C unit, accompanied by N2 abstraction, and reorganization of a dppm subunit and, considered as a whole, to the transformation of the PCP pincer carbodi\u00adphospho\u00adrane system to a phospho\u00adrus ylide ligand. After removal of the halogenides, the iridium center is stabilized by the carbonyl O atom through the formation of a five-membered chelate ring. A PCO pincer ligand system is thereby generated, which coordinates the iridium(III) atom threefold in a facial manner. The addition of carbon monoxide causes a replacement of the carbonyl O atom of the acetate subunit by a carbonyl ligand.The reaction of Cl with ethyl diazo\u00adacetate, a well known C=C coupling reagent, leads to the formation of a C=C unit, accompanied by N2 abstraction, reorganization of a dppm subunit and, considered as a whole, to the transformation of the PCP pincer carbodi\u00adphospho\u00adrane system to a phospho\u00adrus ylide ligand. After removal of the halogenides, the iridium center is stabilized by the carbonyl O atom through the formation of a five-membered chelate ring. A PCO pincer ligand system is thereby generated, which coordinates the iridium(III) atom threefold in a facial manner. The phospho\u00adrus electron-donor atoms and the ylide carbon atom of the resulting (CF3O3S)2 complex, also termed as [bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)methane]({[(di\u00adphenyl\u00adphosphan\u00adyl)meth\u00adyl]di\u00adphenyl\u00adphosphanyl\u00adidene}(eth\u00adoxy\u00adoxoethanyl\u00adidene)methanyl\u00adidene-\u03ba3P,C,O)hydridoiridium(III) bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfonate), are in plane and the hydrido ligand and the carbonyl O atom are located trans to each other, perpendicular to the meridional plane. The addition of carbon monoxide causes a replacement of the carbonyl O atom of the acetate subunit by a carbonyl ligand, thereby creating [bis\u00ad(di\u00adphenyl\u00adphosphan\u00adyl)methane]\u00adcarbon\u00adyl({[(di\u00adphenyl\u00adphosphan\u00adyl)meth\u00adyl]di\u00adphenyl\u00adphosphanyl\u00adidene}(eth\u00adoxy\u00adoxoethanyl\u00adidene)methanyl\u00adidene-\u03ba2P,C}hydridoiridium(III) bis\u00ad(tri\u00adfluoro\u00admethane\u00adsulfonate)\u2013di\u00adchloro\u00admethane\u2013ethyl acetate (6/2/3) or, more simply, (CF3O3S)2\u00b70.33CH2Cl2\u00b70.5C4H8O2. One tri\u00adfluoro\u00admeth\u00adane\u00adsulfonate counter-ion of 3 shows positional disorder in a 2:1 ratio. Complex 4 shows pseudo-merohedral twinning (matrix: The reaction of [Ir Addition of ethyl diazo\u00adacetate (EDA) to Cl complex 1 (Scheme 1) leads to a carbon\u2013carbon coupling reaction via extrusion of a dppm subunit, which is stabilized by two phospho\u00adrus\u2013iridium(III) electron donor\u2013acceptor inter\u00adactions under the formation of a four-membered chelate ring. This reaction sequence, produced by the inter\u00adaction of the doubly ylidic carbon atom with an electrophile containing the extraordinarily good di\u00adnitro\u00adgen withdrawing group, may be described as Wittig-type carbon\u2013carbon coupling reaction, which has rarely been reported in carbodi\u00adphospho\u00adrane chemistry units, the iridium(III) center coordinates a hydrido and a chlorido ligand trans to each other. Reaction of the monocationic Cl complex 2 with two equivalents of thallium(I)tri\u00adfluoro\u00admethane\u00adsulfonate (TfOTl) causes the removal of the chlorido ligand and the chloride counter-ion with concomitant coordination of the acetate carbonyl oxygen atom in a facial manner, resulting in formation of the dicationic (CF3O3S)2 complex 3.Treatment of our PCP pincer ligand system [CH(dppm)et al. (CO)3(dppm)] complex at low temperature generates the corresponding vinyl\u00adidene [MnI(C=CH\u2014CO2Me)(CO)3(dppm)]BF4 complex, which rearranges via insertion of the vinyl\u00adidene ligand into the manganese\u2013phospho\u00adrus bond upon warming to room temperature to an [MnI{(dppm)C=CH(CO2Me)}(CO)3]BF4 complex. Exposure of complex 3 to carbon monoxide gas cleaves the iridium(III) carbonyl oxygen bond under coordination of a carbonyl ligand. Up to now, we have been unable to obtain suitable single crystals of complexes 1 and 2; however, it proved possible to crystallize the (CF3O3S)2 (3) and (CF3O3S)2 (4) products, the latter as a mixed di\u00adchloro\u00admethane\u2013ethyl acetate solvate.A similar ligand arrangement in the coordination sphere of manganese(I) was previously mentioned by Ruiz al. 2005. Protona3 reveal an ortho\u00adrhom\u00adbic crystal system in space group P212121 centre is coordinated in a facial mode by the PCO pincer ligand system via a phosphine functionality, an ylidic carbon atom and a carbonyl oxygen atom of the ester group. The coordination sphere of the iridium(III) atom is completed by one bidentate dppm and one hydrido ligand. Furthermore, all phospho\u00adrus atoms and the iridium atom are positioned in a common plane and the carbonyl oxygen atom as well as the hydride anion are located perpendicularly to this plane, trans to each other. The iridium center creates with its coordination sphere a distorted octa\u00adhedral geometry and the deviations are caused by the presence of the strained four-membered dppm chelate ring [P3\u2014Ir1\u2014P4 = 70.2\u2005(1)\u00b0] and the tridentate PCO ligand . A C1\u2014C4 distance of 1.335\u2005(9)\u2005\u00c5 indicates a double bond and the sum of angles of 358.3\u00b0 permits the designation of the C1 surroundings as a planar surface. The Ir1\u2014O1 bond length of 2.239\u2005(4)\u2005\u00c5 is close to the value [2.262\u2005(4)\u2005\u00c5] in the related [Ir(CO2CH3){C(CO2CH3)CH(CO2CH3)}(PPh3)(2-Ph2PC6H4NH)] complex complex, which is stabilized by two tri\u00adfluoro\u00admethane\u00adsulfonate counter-ions metal atom shows a distorted octa\u00adhedral geometry. It coordinates a dppm unit and a PCO pincer ligand system in a meridional manner and, perpendicular to this plane, a hydrido ligand. The only difference is that the carbonyl oxygen atom of the PCO ligand system is uncoordinated and has been substituted by a carbonyl ligand. The carbonyl ligand reveals relatively long Ir1\u2014C8 [1.965\u2005(15)\u2005\u00c5] and C8\u2014O3 [1.116\u2005(14)\u2005\u00c5] distances, caused by the location trans to the hydrido ligand. Moreover, the substitution results in an overall lengthening of the Ir\u2014P and the Ir\u2014C separations and 3.473\u2005(10)\u2005\u00c5 2-Synthesis of Cl (1): A mixture of 0.1\u2005ml MeCN, 20.4\u2005mg of [CH(dppm)2]Cl (0.0250\u2005mmol) and 8.4\u2005mg of [IrCl(cod)]2 (0.0125\u2005mmol) was stirred for 1\u2005min. The resulting solution contains predominantly the well known Cl2 ]Cl complex 1. 31P {1H} NMR : \u03b4 = 1.5 , 31.5 p.p.m.; 13C {1H} NMR : \u03b4 = \u221228.9 p.p.m.; 1H NMR : \u03b4 = \u221221.3 p.p.m.4H6O2)(dppm)-Synthesis of [Ir{CH]Cl (2): While stirring the aforementioned MeCN solution of Cl2 and Cl (1), 0.26\u2005ml of CHCl3 and 0.24\u2005ml of a solution of ethyl diazo\u00adacetate in CHCl3 were added successively. The Cl2 by-product is slowly transformed to 1, which in turn reacts with ethyl diazo\u00adacetate. After standing for 24\u2005h, product 2 was generated almost qu\u00adanti\u00adtatively. 31P {1H} NMR : \u03b4 = 26.5 , 45.7 , \u221252.5 , \u221236.7 p.p.m.; 13C {1H} NMR : \u03b4 = 139.2 p.p.m.; 1H NMR : \u03b4 = \u221217.7 p.p.m.4H6O2)(dppm)-Synthesis of [Ir{C2 (3):}(dppm)H] tri\u00adfluoro\u00admethane\u00adsulfonate (0.0597\u2005mmol) were dissolved in MeOH (0.1\u2005ml) and added to a solution of complex 2 (0.025\u2005mmol) in chloro\u00adform/aceto\u00adnitrile (5:1). After stirring for 15\u2005min the precipitated TlCl was separated, all solvents were removed and complex 3 was obtained . Single beige\u2013white crystals of complex 3 were grown slowly from acetone (0.5\u2005ml), covered with 0.3\u2005ml of hexane. 31P {1H} NMR : \u03b4 = 9.1 , 36.0 , \u221244.2 , \u221233.1 p.p.m.; 13C {1H} NMR (CDCl3): \u03b4 = 181.4 p.p.m.; 1H NMR (CDCl3): \u03b4 = \u221223.3 p.p.m.4H6O2)(dppm)-Synthesis of [Ir{C2 (4):}(CO)(dppm)H](CF Complex 3 was dissolved in CHCl3 (0.6\u2005ml), the mixture was filtered and the solution was placed in an atmos\u00adphere of CO. After standing for 16\u2005h product 4 was formed and single crystals were obtained via layering of a solution of complex 4 dissolved in CH2Cl2 with EtOAc. 31P {1H} NMR (CHCl3/CH3OH): \u03b4 = 5.3 , 57.1 , \u221255.6 , \u221244. 7 p.p.m.; 13C {1H} NMR (CDCl3/CD3OD): \u03b4 = 134.8 p.p.m.; 1H NMR (CDCl3/CD3OD): \u03b4 = \u22128.8 p.p.m.3 and 4 were processed without absorption corrections. In relation to the structure determination of complex 3, the hydrido ligand was detected and refined isotropically. One tri\u00adfluoro\u00admethane\u00adsulfonate counter-ion shows positional disorder in a 2:1 ratio, caused by an overlying of the C9, O16 and F16 positions. These positions were also refined isotropically. The structure determination of complex 4 resulted in the detection of pseudo-merohedral twinning = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S205698901801455X/hb7757sup1.cifCrystal structure: contains datablock(s) global, 3, 4. DOI: 10.1107/S205698901801455X/hb77573sup2.hklStructure factors: contains datablock(s) 3. DOI: 10.1107/S205698901801455X/hb77574sup4.hklStructure factors: contains datablock(s) 4. DOI: 1849369, 1849368CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E configuration with respect to the C=N double bond. The benzene rings form a dihedral angle of 58.06\u2005(9)\u00b0. In the crystal, the mol\u00adecules are linked by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds into chains, which are further connected into a three-dimensional network by C\u2014H\u22ef\u03c0 inter\u00adactions.The title Schiff base compound displays an 15H13BrN2O2, displays an E configuration with respect to the C=N double bond, which forms a dihedral angle of 58.06\u2005(9)\u00b0 with the benzene ring. In the crystal, the mol\u00adecules are linked into chains parallel to the b axis by N\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds, giving rise to rings with an R21(6) graph-set motif. The chains are further linked into a three-dimensional network by C\u2014H\u22ef\u03c0 inter\u00adactions. A Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from C\u22efH (33.2%), H\u22efH (27.7%), Br\u22efH/H\u22efBr (14.2%) and O\u22efH/H\u22efO (13.6%) inter\u00adactions. The title compound has also been characterized by frontier mol\u00adecular orbital analysis.The title Schiff base compound, C Schiff bases are of great importance in the field of coordination chemistry because they are able to form stable complexes with metal ions -4-bromo-N\u2032-(4-meth\u00adoxy\u00adbenzyl\u00adidene)benzohydrazide is reported.Schiff bases are nitro\u00adgen-containing compounds that were first obtained by the condensation reactions of aromatic amines and aldehydes \u2005\u00c5] and C7=O2 [1.222\u2005(3)\u2005\u00c5] bond lengths confirm their double-bond character. The C7\u2014N1, N1\u2014N2 and C3\u2014Br1 bond lengths are 1.354\u2005(3), 1.379\u2005(3) and 1.894\u2005(3)\u2005\u00c5, respectively. The central O2/C7/N1/N2 fragment is approximately planar (r.m.s. deviation 0.0141\u2005\u00c5) and forms dihedral angles of 32.5\u2005(2) and 27.2\u2005(2)\u00b0 with the C1\u2013C6 and C9\u2013C14 rings, respectively. The dihedral angle formed by the aromatic rings is 58.06\u2005(9)\u00b0.The asymmetric unit of the title compound Fig.\u00a01 consistsb-axis direction by N1\u2014H1N\u22efO2 and C8\u2014H8\u22efO2 hydrogen-bonding inter\u00adactions acts as an electron donor and the LUMO acts as an electron acceptor. If the energy gap is small then the mol\u00adecule is highly polarizable and has high chemical reactivity. The energy levels were computed by the DFT-B3LYP/6-311G++ method benzohydrazide benzohydrazide benzohydrazide benzohydrazide methyl\u00adene)benzohydrazide methyl\u00adidenebenzohydrazine of 4-bromo\u00adbenzohydrazide (0.213\u2005mg) and a hot ethano\u00adlic solution of 4-meth\u00adoxy\u00adbenzaldehyde (0.136\u2005mg). The mixture was refluxed for 8\u2005h, then it was cooled and kept at room temperature. The powder formed was recrystallized from DMSO. Colourless block-shaped crystals suitable for X-ray analysis were obtained after a few days on slow evaporation of the solvent.Uiso(H) = 1.2Ueq or 1.5Ueq(C) for methyl H atoms. A rotating model was used for the methyl H atoms. Three outliers global, I, 1. DOI: 10.1107/S2056989018013373/rz5241Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018013373/rz5241Isup3.cmlSupporting information file. DOI: 1587248CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The coordination sphere of each Sn atom is best described as a distorted trigonal bipyramid. Each stannic ion in the complex is in a C2O2N environment. The two homologous parts of the doubly deprotonated ligand are located in trans positions with respect to the C\u2014C bond of the oxalamide group. The oxalamide group exhibits an asymmetric coordination geometry, as seen by the slight difference between the C\u2014O and C\u2014N bond lengths. The three-dimensional network is a multilayer of complex mol\u00adecules with no strong supramolecular inter\u00adactions.The binuclear complex, [Sn The stannic units are connected by the doubly deprotonated ligand which play a bridging role in a trans conformation. Each stannic unit is coordinated to the ligand via an imino\u00adlate O atom, a phenolate O atom and an imine N atom. Each Sn atom is penta\u00adcoordinated. The Sn\u2014C bond lengths [2.158\u2005(3)\u20132.168\u2005(3)\u2005\u00c5] are slightly shorter than the values reported for complexes containing the [Sn(tBu)2]2+ unit and 2.0979\u2005(18)\u2005\u00c5, respectively, for Sn1 and Sn2] are shorter than the Sn\u2014Oimino\u00adlate bond lengths (Table\u00a01phenolate distances associated with the strong coordination [1.302\u2005(3)\u20131.308\u2005(3)\u2005\u00c5] are longer than the C\u2014Oimino\u00adlate bonds associated with the less strong coordination [1.283\u2005(3)\u20131.288\u2005(3)\u2005\u00c5]. The coordination sphere SnNC2O2 for each of the two Sn atoms can be characterized by the trigonality parameter \u03c4 = (\u03b2 \u2212 \u03b1)/60, with \u03b1 and \u03b2 being the two largest angles around Sn \u00b0 for Sn1 and 130.02\u2005(12)\u00b0 for Sn2] result in compression of the bond angles with the third atom which forms the equatorial plane with the two tert-butyl groups . The sum of the angles in the basal planes are, respectively, 359.99\u00b0 for Sn1 and 359.94\u00b0 for Sn2. The O atoms occupy the apical positions with comparable angles of 154.61\u2005(7)\u00b0 for Sn1 and 154.73\u2005(7)\u00b0 for Sn2. The angles between the apical O atoms and the atoms in the basal plane are in the range 72.35\u2005(7)\u201397.12\u2005(11)\u00b0 for Sn1 and between 72.39\u2005(6) and 96.48\u2005(9)\u00b0 for Sn2. The ligand, which acts in a tridentate fashion, forms two rings upon coordination with the tin centres, i.e. a five-membered OCNNSn ring and a six-membered OCCCNSn ring, sharing atom N1 for Sn1 and N4 for Sn2. The angles resulting from the five-membered ring [N1\u2014Sn1\u2014O2 = 72.35\u2005(7)\u00b0 and N4\u2014Sn2\u2014O3 = 72.39\u2005(6)\u00b0] are much smaller than the angles resulting from the six-membered ring [N1\u2014Sn1\u2014O1 = 82.32\u2005(8)\u00b0 and N4\u2014Sn2\u2014O4 = 82.39\u2005(7)\u00b0]. The better flexibility of the six-membered ring can explain this observed difference in values. The five- and six-membered rings obtained after coordination of the ligand are not planar, as indicated by the torsion angles for the two Sn atoms in the complex: Sn1\u2014N1\u2014N2\u2014C8 0.6, Sn1\u2014O2\u2014C8\u2014N2 0.5, Sn1\u2014O1\u2014C1\u2014C6 6.3, Sn1\u2014N1\u2014C7\u2014C6\u00a0\u2212\u00a02, Sn2\u2014N4\u2014N3\u2014C9 2.1, Sn2\u2014O3\u2014C9\u2014N3\u00a0\u2212\u00a01.2, Sn2\u2014O4\u2014C16\u2014C11\u00a0\u2212\u00a03.7 and Sn2\u2014N4\u2014C10\u2014C11\u00a0\u2212\u00a00.5\u00b0. For all four tBu groups, the angles around the central C atom (Sn\u2014C\u2014C and C\u2014C\u2014C) vary in the range from 106.0\u2005(3) to 112.3\u2005(4)\u00b0 and indicate a tetra\u00adhedral environment around the central C atom. Both tBu groups reveal an eclipsed conformation regarding the methyl groups. The C\u2014C bond lengths are in the range 1.81\u2005(5)\u20131.542\u2005(9)\u2005\u00c5 and are comparable to the values found in the literature was added a solution of salicyl\u00adaldehyde (2\u2005mmol) in 10\u2005ml of the same mixture. A white precipitate appeared and the resulting mixture was stirred at room temperature for 24\u2005h. The suspension was filtered and the solid was washed with 2 \u00d7 10\u2005ml of water and 2 \u00d7 10\u2005ml of ether. The solid was recrystallized from a mixture of chloro\u00adform and methanol (1:1 v/v). The white powder collected was dried under P2O5. Yield 90% (H4L). Calculated for C16H14N4O4: C 58.89, H 4.32, N 17.17%; found: C 59.02, H 4.37, N 17.24%. IR (cm\u22121): 3277 (\u03bd O\u2014H), 1664 (\u03bd C=O), 1601 (\u03bd C=N), 1533, 1486, 1457, 1357, 1304, 1259, 1218, 1161 (\u03bd C\u2014O), 776, 673. 1H NMR: \u03b4 12.6 , 11.00 , 8.85 , 7.6\u20137.00 . 13C NMR: \u03b4 158.5, 156.8, 151.98, 148.00, 132.93, 130.27, 120.37, 119.54, 117.39. To a mixture of H4L (2\u2005mmol) and tri\u00adethyl\u00adamine (4\u2005mmol) in 10\u2005ml of ethanol was added SnCl2tBu2 (2\u2005mmol) in ethanol (10\u2005ml). The resulting yellow mixture was stirred under reflux for 120\u2005min and the resulting brown solution was filtered. The filtrate was kept at 298\u2005K and after one week yellow crystals suitable for X-ray analysis appeared and were collected by filtration. Yield 40%, m.p. 243\u00b0C. Calculated for C32H46N4Sn2O4: C 48.77, H 5.88, N 7.11%; found: C 48.64, H 5.96, N 7.09%. IR (cm\u22121): 1609, 1537, 1516, 1468, 1441, 1367, 1310, 1275, 1198, 1167, 1150, 870, 771, 754. 1H NMR: \u03b4 8.85 ; 7.13\u20136.69 ; 1.33 . 13C NMR: \u03b4 168.80, 163.68, 135.85, 134.72, 122.22, 116.99, 41.53, 29.96.To a solution of oxalyldihydrazine (1\u2005mmol) in a mixture of water and methanol (1:3 Uiso(H) = 1.2Ueq(C) (1.5 for CH3 groups).Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989018007077/ex2008sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018007077/ex2008Isup2.hklStructure factors: contains datablock(s) I. DOI: 1842349CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Two oxetanocin A derivatives have been synthesized stereospecifically. The crystal structures of both derivatives and a synthetic inter\u00admediate are reported on herein. 10H9ClN4O, 3-[(6-chloro-7H-purin-7-yl)meth\u00adyl]cyclo\u00adbutan-1-one (I), and two N-7 and N-9 regioisomeric oxetanocin nucleoside analogs, C10H13ClN4O, 3-[meth\u00adyl]cyclo\u00adbutan-1-ol (II) and C10H11ClN4O, 3-[(6-chloro-9H-purin-9-yl)meth\u00adyl]cyclo\u00adbutan-1-ol (IV), are reported. The crystal structures of the nucleoside analogs confirmed the reduction of the N-7- and N-9-substituted cyclo\u00adbutano\u00adnes with LiAl(OtBu)3 to occur with facial selectivity, yielding cis-nucleosides analogs similar to those found in nature. Reduction of the purine ring of the N-7 cyclo\u00adbutanone to a di\u00adhydro\u00adpurine was observed for compound (II) but not for the purine ring of the N-9 cyclo\u00adbutanone on formation of compound (IV). In the crystal of (I), mol\u00adecules are linked by a weak Cl\u22efO inter\u00adaction, forming a 21 helix along [010]. The helices are linked by offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.498\u2005(1)\u2005\u00c5], forming layers parallel to (101). In the crystal of (II), mol\u00adecules are linked by pairs of O\u2014H\u22efN hydrogen bonds, forming inversion dimers with an R22(8) ring motif. The dimers are linked by O\u2014H\u22efN hydrogen bonds, forming chains along [001], which in turn are linked by C\u2014H\u22ef\u03c0 and offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.509\u2005(1)\u2005\u00c5], forming slabs parallel to the ac plane. In the crystal of (IV), mol\u00adecules are linked by O\u2014H\u22efN hydrogen bonds, forming chains along [101]. The chains are linked by C\u2014H\u22efN and C\u2014H\u22efO hydrogen bonds and C\u2014H\u22ef\u03c0 and offset \u03c0\u2013\u03c0 inter\u00adactions [inter\u00adcentroid distance = 3.364\u2005(1)\u2005\u00c5], forming a supra\u00admolecular framework.The crystal structures of an inter\u00admediate, C The mean plane of the cyclo\u00adbutane ring (A = C2\u2032\u2013C5\u2032) is inclined to the mean plane of the purine ring system (B = N1/N37N7/N9/C2/C4/C5/C6/C8) by 52.62\u2005(11)\u00b0, while the torsion angle N7\u2014C5\u2032\u2014C4\u2032\u22efC2\u2032 is ca 125.4\u00b0.In compound (I)tert-but\u00adoxy\u00adaluminum hydride lead to the formation of the oxetanocin derivative compound (II)A) is inclined to the mean plane of the purine ring system (B) by 26.37\u2005(15)\u00b0, while the torsion angle N7\u2014C5\u2032\u2014C4\u2032\u22efC2\u2032 is ca 120.0\u00b0. Atoms C6\u2032 and C4\u2032 are positionally disordered and were split giving a refined occupancy ratio for C6\u2032:C6\u2032B and C4\u2032:C4\u2032B of 0.858\u2005(4):0.142\u2005(4) 4) Fig.\u00a02.trans positioning of the cyclo\u00adbutanol unit, there are no intra\u00admolecular hydrogen bonds between the chlorine atom and the cyclo\u00adbutanol or methyl\u00adene connector as observed in compounds (I)A) is inclined to the mean plane of the purine ring system (B) by 71.20\u2005(13)\u00b0, and the torsion angle N7\u2014C5\u2032\u2014C4\u2032\u22efC2\u2032 is ca 144.8\u00b0.In compound (IV)4 to a di\u00adhydro\u00adpurine . This strained orientation is not observed for the N-9 ketone, hence the integrity of its purine ring is preserved.Reduction of the purine ring of the N-7 cyclo\u00adbutanone to a di\u00adhydro\u00adpurine was observed for compound (II)x\u00a0+\u00a01, y\u00a0\u2212\u00a0z\u00a0+\u00a01 helix along [010], see Fig.\u00a04CgB\u22efCgBi = 3.498\u2005(1)\u2005\u00c5, CgB is the centroid of the purine ring system, \u03b1 = 0.00\u2005(5)\u2005\u00c5, \u03b2 = 21.6\u00b0, inter\u00adplanar distance = 3.252\u2005(1)\u2005\u00c5, offset = 1.289\u2005\u00c5, symmetry code (i) \u2212x\u00a0+\u00a02, \u2212y\u00a0+\u00a01, \u2212z\u00a0+\u00a01.In the crystal of (I)ac plane. In the crystal of (II)f Table\u00a02. The dimf Table\u00a02 and offsCgB\u22efCgBvi = 3.534\u2005(1)\u2005\u00c5, CgB is the centroid of the purine ring system, \u03b1 = 0.02\u2005(10)\u2005\u00c5, \u03b2 = 17.8\u00b0, inter\u00adplanar distance = 3.364\u2005(1)\u2005\u00c5, offset = 1.08\u2005\u00c5, symmetry code (vi) \u2212x\u00a0+\u00a01, \u2212y\u00a0+\u00a01, \u2212z. ]In the crystal of (IV)et al., 2016viz. 6-chloro-9-(3-hy\u00addroxy\u00admethyl-3-hy\u00addroxy\u00adcyclo\u00adbut\u00adyl)purine cyclo\u00adbutanol Potassium carbonate (12.0\u2005mmol) was added to a solution of (3-oxo\u00adcyclo\u00adbut\u00adyl)methyl benzoate (10.0\u2005mmol) in methanol (20\u2005ml) and stirred for 1\u2005h at room temperature. Saturated sodium bicarbonate (10.0\u2005ml) was added and stirring continued for an additional 15\u2005min. The solvent was evaporated under vacuum, followed by purification by flash column chromatography with ethyl acetate, resulting in 3-(hy\u00addroxy\u00admeth\u00adyl)cyclo\u00adbutan-1-one in 70% yield.3-(Hy\u00addroxy\u00admeth\u00adyl)cyclo\u00adbutan-1-one (1\u2005mmol) was dissolved in 10\u2005ml of dry di\u00adchloro\u00admethane and cooled to 195\u2005K. Hunig\u2019s base (3.2\u2005mmol) was added, followed by tri\u00adfluoro\u00admethane\u00adsulfonic anhydride (1\u2005mmol) and the mixture was stirred for 10\u2005min, cooled to 273\u2005K and stirred to obtain the qualitative conversion to (3-oxo\u00adcyclo\u00adbut\u00adyl)methyl tri\u00adfluoro\u00admethane\u00adsulfonate.H-purine (5.61\u2005mmol), potassium hydroxide (5.61\u2005mmol), tris\u00ad[2-(2-meth\u00adoxy\u00adeth\u00adoxy)eth\u00adyl]amine (0.28\u2005mmol), magnesium sulfate (2\u2005g) and anhydrous aceto\u00adnitrile (100\u2005ml), which was then heated to 333\u2005K for 5\u2005h and cooled to room temperature. The product was purified using 5% methanol and 5% tri\u00admethyl\u00adamine in chloro\u00adform, which yielded two UV-active compounds.The (3-oxo\u00adcyclo\u00adbut\u00adyl)methyl tri\u00adfluoro\u00admethane\u00adsulfonate (5.61\u2005mmol) was added to a mixture containing 6-chloro-7H-purin-9-yl)meth\u00adyl]cyclo\u00adbutan-1-one (III) and 37% of the N-7 alkyl\u00adated deriv\u00adative 3-[(6-chloro-7H-purin-7-yl)meth\u00adyl]cyclo\u00adbutan-1-one (I)The two UV-active compounds were separated using flash column chromatography with ethyl acetate, giving 51% of the N-9 alkyl\u00adated derivative; 3-[meth\u00adyl]cyclo\u00adbutan-1-ol (II) 3-[(6-Chloro-7H-purin-7-yl)meth\u00adyl]cyclo\u00adbutan-1-one (I)tert-but\u00adoxy\u00adaluminum hydride was added. The mixture was cooled to room temperature and sodium borohydride (0.32\u2005mmol) was added and the resulting mixture allowed to stir overnight. Methanol (2\u2005ml) was added and the mixture allowed to stir overnight to convert the over-reduced 3-[(6-chloro-7H-purin-7-yl]meth\u00adyl)cyclo\u00adbutanone(I) to 3-[meth\u00adyl]cyclo\u00adbutan-1-ol (II)Synthesis ofcis-3-[(6-chloro-9H-purin-9-yl)meth\u00adyl]cyclo\u00adbutan-1-ol (IV) 3-[(6-Chloro-9H-purin-9-yl)meth\u00adyl]cyclo\u00adbutan-1-one (III) (0.21\u2005mmol) was added to diethyl ether and cooled to 195\u2005K and lithium tri-tert-but\u00adoxy\u00adaluminum hydride (0.32\u2005mmol) was added. The reaction was allowed to warm to room temperature and left to stir overnight, which provided qu\u00adanti\u00adtative conversion to cis-3-[(6-chloro-9H-purin-9-yl)meth\u00adyl]cyclo\u00adbutan-1-ol (IV)Pale-yellow plate-like crystals of (I)Uiso(H) = 1.2Ueq(C). For compound (II)B and C4\u2032:C4\u2032B of 0.858\u2005(4):0.142\u2005(4). For the final refinement of compound (II)Crystal data, data collection and structure refinement details are summarized in Table\u00a0510.1107/S2056989019004432/zp2033sup1.cifCrystal structure: contains datablock(s) global, I, II, IV. DOI: 10.1107/S2056989019004432/zp2033Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019004432/zp2033IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989019004432/zp2033IIsup5.cmlSupporting information file. DOI: 1907200, 1907199, 1907198CCDC references: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "E. In the crystal, O\u2014H\u22efO hydrogen-bond inter\u00adactions consolidate the crystal packing. A Hirshfeld surface analysis and fingerprint plots were used to further investigate the inter\u00admolecular inter\u00adactions in the solid state.In the title Schiff base derivative carrying a 2-bromo-3-methyl\u00adphenyl group, the conformation about the C=N bond is 14H12BrNO2, was synthesized by the condensation reaction of 2,3-di\u00adhydroxy\u00adbenzaldehyde and 2-bromo-3-methyl\u00adaniline. It crystallizes in the centrosymmetric triclinic space group PE. The dihedral angle between the planes of the 5-(2-bromo-3-methyl\u00adphenyl ring and the catechol ring is 2.80\u2005(17)\u00b0. In the crystal, O\u2014H\u22efO hydrogen-bond inter\u00adactions consolidate the crystal packing.The title compound, C Schiff PZ = 4 (Z\u2032 = 2). The two crystallographically independent mol\u00adecules have nearly the same geometrical parameters and the primary difference between them is the rotational orientation of H2 and H4A. The discussion will therefore be limited to that of the mol\u00adecule containing O1. The mol\u00adecular structure is constructed from two individually planar rings. The whole mol\u00adecule is approximately planar, with a maximum deviation of 0.117\u2005(3)\u2005\u00c5 from planarity for the hydroxyl O1 atom of the catechol ring. The dihedral angle between the two benzene ring planes is 2.80\u2005(17)\u00b0. The methyl C1 atom deviates from the plane of the C2\u2013C7 benzene ring by 0.039\u2005(2)\u2005\u00c5 while C9 deviates from the plane of the C9\u2013C14 benzene ring by 0.024\u2005(3)\u2005\u00c5. The C8\u2014N1\u2014C7\u2014C6 and C14\u2014 C9\u2014C8\u2014N1 torsion angles are \u22121.6\u2005(5) and \u22121.1\u2005(5)\u00b0, respectively. The planar mol\u00adecular conformation of each molecule is stabilized by an intra\u00admolecular O\u2014H\u22efN hydrogen bond and Cg3\u22efCg4, where Cg1, Cg2, Cg3 and Cg4 are the centroids of the C2\u2013C7, C9\u2013C14, C16\u2013C21 and C23\u2013C28 rings, respectively.In the crystal, the Schiff base units are linked by O\u2014H\u22efO and C\u2014H\u22efO hydrogen bonds -N-(2-bromo\u00adphen\u00adyl)-1-phenyl\u00admethanimine skeleton yielded nine hits. The N1\u2014C8 bond in the title structure is the same length within standard uncertainties as those in the structures of 2-bromo-N-salicylideneaniline -1-(2-fluoro\u00adphen\u00adyl)methanimine -\u00admeth\u00adyl]-4-bromo\u00adphenol naphthaldimine -2-bromo\u00adaniline plot with de and di for the title compound is shown in Fig.\u00a03de = di = 1.5\u2005\u00c5; Fig.\u00a04Hirshfeld surface analysis of the title compound was performed utilizing the A mixture of 2,3-di\u00adhydroxy\u00adbenzaldehyde and 2-bromo-3-methyl\u00adaniline was stirred with ethanol (30\u2005mL) at 377\u2005K for 5\u2005h, affording the title compound . Single crystals suitable for X-ray measurements were obtained by recrystallization from ethanol at room temperature.Uiso(H) = 1.5Ueq(O). The C-bound H atoms were positioned geometrically and refined using a riding model: C\u2014H = 0.93\u2005\u00c5 with Uiso(H) = 1.2Ueq(C) for aromatic H atoms and C\u2014H = 0.96\u2005\u00c5 with Uiso(H) = 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table\u00a0210.1107/S2056989019015718/mw2151sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019015718/mw2151Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019015718/mw2151Isup3.cmlSupporting information file. DOI: 1967023CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "The asymmetric unit of the title solvate comprises a half mol\u00adecule of each component as both species are disposed about a centre of inversion. In the crystal, two-dimensional arrays are formed by amide-N\u2014H\u22efN(pyrid\u00adyl) hydrogen bonds, which are connected into a three-dimensional architecture by C\u2014H\u22ef\u03c0(benzene and pyrid\u00adyl) inter\u00adactions with benzene acting as the acceptor and donor, respectively. 14H14N4O2\u00b7C6H6 , comprises a half mol\u00adecule of each constituent as each is disposed about a centre of inversion. In the oxalamide mol\u00adecule, the central C2N2O2 atoms are planar (r.m.s. deviation = 0.0006\u2005\u00c5). An intra\u00admolecular amide-N\u2014H\u22efO(amide) hydrogen bond is evident, which gives rise to an S(5) loop. Overall, the mol\u00adecule adopts an anti\u00adperiplanar disposition of the pyridyl rings, and an orthogonal relationship is evident between the central plane and each terminal pyridyl ring [dihedral angle = 86.89\u2005(3)\u00b0]. In the crystal, supra\u00admolecular layers parallel to (10via methyl\u00adene-C\u2014H\u22ef\u03c0(benzene) and benzene-C\u2014H\u22ef\u03c0(pyrid\u00adyl) inter\u00adactions. The specified contacts are indicated in an analysis of the calculated Hirshfeld surfaces. The energy of stabilization provided by the conventional hydrogen bonding is just over double that by the C\u2014H\u22ef\u03c0 contacts (dispersion forces).The asymmetric unit of the title 1:1 solvate, C The first of these arises from amide-N\u2014H\u22efO(amide) hydrogen bonding between the amide groups, on both sides of the 3LH2 mol\u00adecule, through ten-membered amide synthons {\u22efHNC2O}2 . Mol\u00adecules of nLH2 also featured prominently in early, systematic studies of halogen bonding. An illustrative example is found in the 1:1 co-crystal formed between 3LH2 and 1,4-di-iodo\u00adbuta-1,3-diyne, I\u2014C\u2261C\u2014C\u2261C\u2014C\u2014I hydrogen bonding and these are linked into a two-dimensional array via amide-N\u2013H\u22efN(pyrid\u00adyl) hydrogen bonds hydrogen bonds to generate a two-dimensional array. In this context, and in the context of recent work on 4LH2 in co-crystals via an analysis of the calculated Hirshfeld surfaces as well as a computational chemistry study.With a combination of centrally located amide and terminal pyridyl functional groups, the isomeric mol\u00adecules related to the title compound of the general formula \u2005\u00c5 to either side of the plane. An intra\u00admolecular amide-N\u2014H\u22efO(amide)i hydrogen bond, occurring between the symmetry related amide groups, gives rise to an S(5) loop, Table\u00a01x, 1\u00a0\u2212\u00a0y, \u2212 z. The crystallographic symmetry also implies an anti\u00adperiplanar disposition of the pyridyl rings. The dihedral angle between the central plane and terminal pyridyl ring is 86.89\u2005(3)\u00b0, indicating an orthogonal relationship.The title co-crystal (I)a), with dimensions defined by O10\u22efO10 and N8\u22efN8 separations of 9.6770\u2005(11) and 12.3255\u2005(11)\u2005\u00c5, respectively. The other notable contacts in the crystal are of the type C\u2014H\u22ef\u03c0, Table\u00a01i.e. two (as acceptor) and two (as donor), such inter\u00adactions, Fig.\u00a02b). The side-on view of Fig.\u00a02b) shown in Fig.\u00a02c) indicates the amide-N\u2014H and pyridyl-N project in all directions around the five-mol\u00adecule aggregate. Indeed, it is the C\u2014H\u22ef\u03c0 inter\u00adactions that connect the layers into a three-dimensional architecture, Fig.\u00a02d).The geometric parameters characterizing the inter\u00adatomic contacts identified in the crystal of (I)Mercury dnorm map of 4LH2 displays several red spots, that range from intense to weak, which reflect the inter\u00adactions identified in the crystal inter\u00adactions, Fig.\u00a04a), with both indicative of contact distances shorter than the respective sum of the van der Waals radii. Reflecting the relatively long separation, the benzene-C11\u2014H11\u22ef\u03c0(pyrid\u00adyl) inter\u00adaction is reflected as only a white spot as the contact distance is only just within the sum of van der Waals radii, as shown in Fig.\u00a04b).The Hirshfeld surface mapped over B\u22ef\u03c0(benzene) contact is indeed electrostatic in nature as revealed by the distinct blue (i.e. electropositive) and red (i.e. electronegative) colour scheme on the surface of the contact points, Fig.\u00a05a). In contrast, the benzene-C11\u2014H11\u22ef\u03c0(pyrid\u00adyl) contact displays pale colouration around the contact zone suggesting that the inter\u00adaction could be attributed to weak dispersion forces, Fig.\u00a05b).The C\u2014H\u22ef\u03c0 inter\u00adactions were subjected to electrostatic potential mapping for verification purposes. The result shows that the methyl\u00adene-C7\u2014H7a)\u2013(e). As shown in the overall fingerprint plot in Fig.\u00a06a), (I)4LH2 mol\u00adecule, therefore indicating that the inter\u00admolecular inter\u00adactions in (I)4LH2 mol\u00adecules. Decomposition of the overall fingerprint plots of (I)di + de \u223c2.42\u2005\u00c5), H\u22efC/C\u22efH , H\u22efO/O\u22efH , H\u22efN/N\u22efH and other contacts (0.8%). Except for the H\u22efH contacts, to differing extents, the remaining major contacts are shorter than the corresponding sum of van der Waals radii for H\u22efC (\u223c2.90\u2005\u00c5), H\u22efO (\u223c2.72\u2005\u00c5) and H\u22efN (\u223c2.75\u2005\u00c5).The two-dimensional fingerprint plots were generated for overall (I)4LH2 mol\u00adecule exhibits at similar distribution of the major contacts compared to overall (I)4LH2 these are either inclined towards the external or inter\u00adnal contacts presumably due to inter\u00adaction with the solvent benzene mol\u00adecule. For instance, the H\u22efC/C\u22efH contact in the individual 4LH2 mol\u00adecule comprises 9.9% -H\u22efC- and 14.6% -C\u22efH- contacts as compared to 12.0 and 14.6% for the equivalent contacts in overall (I)c). Similar observations pertain for the H\u22efO/ O\u22efH and H\u22efN/ N\u22efH inter\u00adactions, Fig.\u00a06d)\u2013(e).The individual i.e. 20.5% for -H\u22efC- and 21.4% for -C\u22efH- contacts. In addition, the solvent mol\u00adecules are sustained in the mol\u00adecular architecture through minor contributions from H\u22efO (5.6%) and H\u22efN (5.9%) contacts, respectively. These inter\u00adactions are at distances of \u223c2.52\u2005\u00c5 (H\u22efH), \u223c2.92\u2005\u00c5 (H\u22efC/C\u22efH), \u223c2.98\u2005\u00c5 (H\u22efO) and \u223c2.79\u2005\u00c5 (H\u22efN), which are greater than the corresponding sum of van der Waals radii, indicating the identified C\u2014H\u22ef\u03c0(benzene and pyrid\u00adyl) inter\u00adactions can largely be considered as localized inter\u00adactions.As for the benzene mol\u00adecule, an irregular fingerprint profile is noted with the distribution dominated by H\u22efH (46.4%) and H\u22efC/ C\u22efH (41.9%) surface contacts. The latter are almost equally distributed between the inter\u00adnal and external contacts, Crystal Explorer 17 based on the procedures as described previously of \u221238.1\u2005kJ\u2005mol\u22121, Table\u00a02B\u22ef\u03c0(benzene) and benzene-C11\u2014H11\u22ef\u03c0(pyrid\u00adyl) contacts with a very similar Eint values of \u221218.9 and \u221216.9\u2005kJ\u2005mol\u22121, respectively, despite the dnorm contact distance being significantly greater for the latter. The calculation results reveal that the repulsion energy is greater in methyl\u00adene-C7\u2014H7B\u22ef\u03c0(benzene) compared with the benzene-C11\u2014H11\u22ef\u03c0(pyrid\u00adyl) contact, which contributes to the slight variation in their Eint values. In short, the N\u2014H\u22efN inter\u00adaction is stabilized largely by electrostatic forces while the C\u2014H\u22ef\u03c0 inter\u00adactions are stabilized largely by dispersion forces. Overall, the crystal of (I)a)\u2013(c).The calculation of inter\u00adaction energy was performed using 4LH2 available in the literature ; molecular inversions were enabled during calculation. The result shows that out of the 20 mol\u00adecules in the cluster, only one 4LH2 mol\u00adecule in each polymorph resembled the reference packing in (I)a) and (b). The result clearly demonstrates the influence of solvent mol\u00adecule upon the mol\u00adecular packing in (I)Calculations were also performed to compare the mol\u00adecular packing similarity of (I)4LH2 exhibit a close similarity in the distribution of mol\u00adecular contacts as judged from the percentage contribution of the corresponding contacts on the Hirshfeld surface. The maximum variation in the distribution of H\u22efH, H\u22efC/C\u22efH, H\u22efO/O\u22efH and H\u22efN/N\u22efH contacts ranged from 7.1, 4.9, 2.2 and 3.8%, respectively among the three crystals.Finally, and referring to Fig.\u00a09Chemical Context, there are two polymorphs available for 4LH2 CH2N(H)C(=S)C(=S)N(H)CH2(C5H4N-n), for n = 2, 3 and 4 oxalamide, was prepared in accordance with the literature procedure \u22121): 3322 \u03bd(N\u2014H), 3141\u20132804 \u03bd(C\u2014H), 1696\u20131661 \u03bd(C=O), 1563\u20131515 \u03bd(C=C), 1414 \u03bd(C\u2014N), 794 \u03b4(C=C).The precursor, Uiso(H) set to 1.2Ueq(C). The nitro\u00adgen-bound H atom was located from difference-Fourier maps and refined with N\u2014H = 0.88\u00b10.01\u2005\u00c5, and with Uiso(H) set to 1.2Ueq(N).Crystal data, data collection and structure refinement details are summarized in Table\u00a0410.1107/S2056989019009551/hb7835sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989019009551/hb7835Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019009551/hb7835Isup3.cmlSupporting information file. DOI: 1938031CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Mature RNA (miRNeasy Mini kit) was reverse transcribed using RT2 First strand cDNA synthesis kit. The cDNA was used on the real\u2010time RT2 Profiler PCR Array in combination with RT2 SYBR\u00ae Green qPCR Mastermix (Roche LightCycler\u00ae 480 Instrument). Threshold cycle (CT) values (excel file) were uploaded onto the data analysis centre web portal (http://www.qiagen.com/geneglobe). CT values were normalized based on a Manual Selection of reference genes. The fold change/regulation (2^(\u2010\u0394\u0394CT)) was calculated using \u0394\u0394CT method .The isolated hVFs were grouped into fibroblasts\u2010less differentiated (HF\u2010LD) and fibroblasts\u2010highly differentiated (HF\u2010HD) based on their \u03b1\u2010SMA expression (immunoblot), compared to the control hVFs Figure A,B. Poly2.2TFAP2A\u2014F:5\u2032\u2010GACCTCTCGATCCACTCCTTAC\u20103\u2032 R: 5\u2032\u2010GAGACGGCATTGCTGTTGGACT\u20103\u2032; \u03b2\u20102\u2010microglobulin (B2M)\u2010 F: 5\u2032\u2010CCACTGAAAAAGATGAGTATGCCT\u20103\u2032 and R: 5\u2032\u2010CCAATCCAAATGCGGCATCTTCA\u20103\u2032. The following PrimeTime qPCR mouse primer assays were used: \u03b1\u2010SMA (Mm.PT.58.16320644); COL1A1 (Mm.PT.58.7562513); COL2A1 (Mm.PT.58.5206680); COL3A1 (Mm.PT.58.13848686), TGFBR1 (Mm.PT.58.28402453), TGFBR2 (Mm.PT.58.6358355) and B2M (Mm.PT.39a.22214835). The cycling conditions were 95\u00b0C for 10\u00a0minutes, followed by 40 cycles at 95\u00b0C for 15\u00a0s, 1\u00a0minute at 60\u00b0C, and 72\u00b0C for 40\u00a0s. Melt curve analysis was performed by an additional dissociation step of 1 cycle at 95\u00b0C for 5\u00a0s followed by 65\u00b0C for 1\u00a0min and ramping data collection to 97\u00b0C. Relative expression values (\u0394Ct) were obtained by normalizing Ct values (Roche Lightcycler 480 Software v1.5.1.62) of the tested genes with that of B2M.Total RNA was isolated from hVFs (miRNeasy Mini kit) and reverse transcribed (miScript RT II kit) with the supplied HiFlex buffer. qPCR was performed on the LightCycler 480 Instrument II, using the Power SYBR Green PCR Master Mix and 10\u00a0ng diluted cDNA per well. The following human primers were used: 2.3Creative Biogene, Shirley, NY.The TFAP2A knockout cell line with NIH/3T3 fibroblasts (TFAP2A\u2010KO) was established using CRISPR/CAS9 technology through 2.42 with DMEM media (10% BCS) and incubated at 37\u00b0C under 5%CO2. Following 24\u00a0hours, hVFs were either treated with TGF\u2010\u03b21 (5\u00a0ng/mL), angiotensin II (100\u00a0nM) or kept as control in DMEM media (2.5% BCS). After 48\u201072\u00a0hours, the fibroblasts/myofibroblasts were rinsed with Dulbecco's PBS and assayed.Fibroblasts from wild\u2010type or TFAP2A\u2010KO groups were plated at 4000 cells/cm2.5Standard western protocols were followed2.6t\u00a0\u2212\u00a0t0]/{[log(Nt)\u2010log(N0)]/log(2)}, where t0 refers time , t represents time (second count), N0 refers count at time t0, and Nt represents count at time t.Both WT and KO fibroblasts were plated as stated before in triplicate (per time\u2010point) in 6\u2010well plates and counted by Cellometer Auto 2000 at 24, 48, and 72\u00a0hours post\u2010plating. Doubling time was calculated by [3TFAP2A expression, along with ELK1, was decreased with decrease in differentiation as visualized in the heat map 1A1 is increased in TFAP2A\u2010KO fibroblasts compared to the wild\u2010type and highly differentiated (HF\u2010HD) based on their \u03b1\u2010SMA expression, compared to control hVFs as shown in Figure There is no conflict of interest.GRR initiated, designed, executed, analysed the study and wrote the manuscript; SE executed the real\u2010time PCR and PCR array; CW and PH implemented the cell culture, immunoblotting and proliferation assays; FXD, LE, FR and AJ interpreted data and proof\u2010read the manuscript.\u00a0Click here for additional data file."} diff --git a/PMC_clustering_20.jsonl b/PMC_clustering_20.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..b0d2010d927614edb05ee3f810bd297b29b007cb --- /dev/null +++ b/PMC_clustering_20.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:2b7e745d9c9320e5a5312c5a652d1df27815ef7fea411e6f473a47f7be19811f +size 26368927 diff --git a/PMC_clustering_21.jsonl b/PMC_clustering_21.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..f5e8cbd02adce464882c5bbbf51821bb65239ae5 --- /dev/null +++ b/PMC_clustering_21.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:e364abc5a60f304568c37ede18fe0a772916ca6861064c1f97f44f888b9d2f5e +size 46770626 diff --git a/PMC_clustering_22.jsonl b/PMC_clustering_22.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..d995e9c42d373eac84122d2fb44ce5ed36d85670 --- /dev/null +++ b/PMC_clustering_22.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:123b75b5aeaeed919f689a2f10952b49190f08c970da3e82e54cb027a3c40bc3 +size 33765688 diff --git a/PMC_clustering_23.jsonl b/PMC_clustering_23.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..cceef9f4c5d5d36fba6c55b17edf3e7cf6cd9425 --- /dev/null +++ b/PMC_clustering_23.jsonl @@ -0,0 +1,608 @@ +{"text": "The Vincent Cristofalo Rising Star Award in Aging Research lecture will feature an address by the 2020 recipient, Sean P. Curran, PhD. This award is given by the American Federation for Aging Research, Inc"} +{"text": "In \u201cElectronic Cognitive Screen Technology for Screening Older Adults With Dementia and Mild Cognitive Impairment in a Community Setting: Development and Validation Study\u201d :e17332) the authors noted the need to revise the \u201cAcknowledgments\u201d section.In the originally published article, the Acknowledgments section read as follows:The development of EC-Screen is supported by The Hong Kong Jockey Club Charities Trust. We greatly appreciate the contributions of the Jockey Club Centre for Positive Ageing in the design and development of the EC-Screen, and Mindvivid Limited for program development of EC-Screen. We also thank Ms. Anthea Ng for her help in data collection and entry.This section has been revised to:We thank Professor JE Morley, Saint Louis University School of Medicine, USA, for having agreed to let us adapt his Rapid Cognitive Screen test into EC-Screen for older Chinese people. We are also grateful for the funding support from the Hong Kong Jockey Club Charities trust, and for the support of the Jockey Club Centre for Positive Ageing for its contribution in design and data collection. We appreciate the support from Mindvivid Limited in software development of EC-Screen. We also thank Ms. Anthea Ng, research assistant of the Division of Neurology at The Chinese University of Hong Kong, for her help in data collection and entry.The correction will appear in the online version of the paper on the JMIR Publications website on January 19, 2021, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "In the original article, we neglected to include the funder KideOn Research Group of the Basque Government, Ref.: IT1342-19 (A category).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Jianzhong Li and Shasha Tao. As well as having affiliation(s) existed in the paper, they should also have Medical College of Soochow University, Suzhou, China as the first affiliation.In the published article, there was an error regarding the affiliation for The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Incorrect funding information is included in the Acknowledgments. The correct funding information should be included in the Funding statement and read as follows: The publication was co-financed within the framework of the Ministry of Science and Higher Education program as \"Regional Initiative Excellence\" in years 2019\u20132022 (Project No. 005 / RID / 2018/19). No additional external funding was received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The raw data underlying the findings of this study are missThe following information is missing from the Funding statement: This study was funded by grants to NA from Astex Inc. (GRANT # 120039) and the Van Andel Research Institute\u2013Stand Up To Cancer Epigenetics Dream Team. Stand Up To Cancer is a program of the Entertainment Industry Foundation, administered by AACR.There is information missing from the Competing Interests statement. The correct Competing Interests statement is as follows: The authors of this manuscript have read the journal\u2019s policies and have the following competing interests to declare: NA was the recipient of grants from Astex Inc. and the Van Andel Research Institute. Additionally, NA has served as consultant to Johnson & Johnson, an advisor to Celgene, and a member of the Scientific Advisory Council to the No Stomach for Cancer Foundation. There are no patents or marketed products to declare. This does not affect our adheres to PLOS ONE policies on sharing data and materials.S1 Data(ZIP)Click here for additional data file."} +{"text": "In \u201cImproving Nutrition and Activity Behaviors Using Digital Technology and Tailored Feedback: Protocol for the Tailored Diet and Activity (ToDAy) Randomized Controlled Trial\u201d :e12782), corrections have been made to the article title and to three places in the text to reflect that the term \u201cLiveLighter\u201d is a registered trademark.The original title of this paper was:Improving Nutrition and Activity Behaviors Using Digital Technology and Tailored Feedback: Protocol for the LiveLighter Tailored Diet and Activity (ToDAy) Randomized Controlled TrialThe title has been changed to:Improving Nutrition and Activity Behaviors Using Digital Technology and Tailored Feedback: Protocol for the Tailored Diet and Activity (ToDAy) Randomized Controlled TrialIn the originally published paper, the caption of Table 1 read:Table 1. Frequency of assessment of variables in the LiveLighter ToDAy study for the tailored feedback, active control, and online control groups.The caption of Table 1 has been changed to read:Table 1. Frequency of assessment of variables in the ToDAy study for the tailored feedback, active control, and online control groups.Under the first paragraph of the section \"Process Evaluation\" in the originally published paper, the sentence:Selected program completers and noncompleters (tailored feedback and active control groups) will be invited to participate in one-on-one interviews concerning their perceptions of the LiveLighter ToDAy intervention.has been replaced by the sentence:Selected program completers and noncompleters (tailored feedback and active control groups) will be invited to participate in one-on-one interviews concerning their perceptions of the ToDAy intervention.In the originally published paper, the Acknowledgments section read:Funding for the LiveLighter ToDAy study is provided by a Healthway Health Promotion Research Grant and the East Metropolitan Health Service. The mFR app is funded by NIH-NCI (1U01CA130784-01) and NIH-NIDDK . CC is supported by an NHMRC Senior Research Fellowship and a University of Newcastle \u2013Gladys M Brawn Senior Research Fellowship. Funding for Fitabase is provided by a Curtin Institute of Computation grant. The sponsors had no role in the design of the study; collection, analyses, or interpretation of data; writing of the manuscript; and decision to publish the results.This section has been changed to read:Funding for the ToDAy study is provided by a Healthway Health Promotion Research Grant and the East Metropolitan Health Service. The mFR app is funded by NIH-NCI (1U01CA130784-01) and NIH-NIDDK . CC is supported by an NHMRC Senior Research Fellowship and a University of Newcastle \u2013Gladys M Brawn Senior Research Fellowship. Funding for Fitabase is provided by a Curtin Institute of Computation grant. The sponsors had no role in the design of the study; collection, analyses, or interpretation of data; writing of the manuscript; and decision to publish the results. The term \u201cLiveLighter\u201d is a registered trademark. The LiveLighter campaign is funded by the Western Australian Department of Health and at the time of this study, delivered by the National Heart Foundation (WA Division), and currently delivered by Cancer Council Western Australia.The correction will appear in the online version of the paper on the JMIR Publications website on December 2, 2020, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "In the original article, we neglected to include the funder: Cancer Prevention Research Institute of Texas Training Grant (CPRIT RP170067) to Hannah Savage.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Dear Dr. Yokokura:Congratulations on the launch of JMA Journal by the Japan Medical Association (JMA) and Japanese Association of Medical Sciences (JAMS). As an official English medical journal covering all aspects of medicine and medical care, including original research, health policy, and global health, I am sure it will be a success not only with clinical investigators in Japan, but worldwide. The publication of this type of comprehensive medical journal is the first challenge in Japan. We sincerely hope that the JMA Journal will promote international medical science and contribute to the improvement of the quality of health care by being read and cited by many people.If I or any of my staff can be of any help in this important endeavor please let us know.Good luck with this important initiative of the JMA and JAMS.Sincerely,Howard Bauchner, MDEditor in Chief, JAMA and the JAMA Network (2011-)Senior Vice-President, American Medical Association"} +{"text": "The Editors highlight that the information presented in the original version was provided by the authors at submission and revision. In addition, the authors had the opportunity to correct the information during proofreading, before approving for publication. | Jinglin Wang Correspondence: Haozhen Ren: Acknowledgments: This work was supported by grants from the Nanjing Medical Science and Technique Development Foundation (No. QRX17129), the Certificate of China Postdoctoral Science Foundation Grant (2018M642222), and the Jiangsu Province Natural Science Foundation (BK20190114).*These authors contributed equally to this study."} +{"text": "It is with great grief that we announce the loss of our friend and mentor, Professor Dr. Ovidiu Alexandru B\u0103jenaru. It would be difficult to write of our beloved colleague in these pages without betraying emotion. A brilliant neurologist, researcher, teacher, husband, father, he leaves behind a profound legacy of perhaps the most influential contemporary figure in Romanian neurology.We remember and cherish some of his most significant achievements over his long and fruitful career. Professor B\u0103jenaru is the founding President of the Romanian Society of Neurology (RSN), an organization that he has formally led for 12 years. In this position, he has contributed decisively to the advancement of science and education in our field. He spearheaded the development of a vision for clinical neurology in our country, based on the conceptual changes determined by colossal advances in neuroscience and modern medical technologies, and integrated using traditional clinical training.Professor Ovidiu B\u0103jenaru represented Romania at top-notch medical forums. He was Romania\u2019s official representative at the World Federation of Neurology (since 2001), the European Stroke Organization, and the European Union of Medical Specialists (Board of Neurology permanent representative of Romania since 2008). Dr. B\u0103jenaru was also a corresponding member of the Romanian Academy (since 2016), as well as a member of the Romanian Group of Brain Research (since 2016), member in the Bureau of Medical Services (since 2018), member of the Academy of Medical Sciences (since 2013).His primary areas of interest were neurodegenerative diseases (particularly Parkinson\u2019s disease and Alzheimer\u2019s disease), muscular dystonia and other motor behavior abnormalities, cerebrovascular diseases, multiple sclerosis, and sleep pathology in neurological disorders and medical conditions \u2013 all fields in which he became a leading authority. His scientific activity includes over 500 presentations at national and international events and over 50 international multicentric clinical trials as a principal investigator.Professor B\u0103jenaru coordinated the interdisciplinary medical group for the treatment by deep brain stimulation for Parkinson\u2019s disease from the Bucharest Emergency University Hospital (since 2006), the first Romanian medical group for the interventional treatment of epilepsy (since 2009), and the national health programs for neurological disorders of the National Health Insurance House and the Romanian Ministry of Health (since 2014). Between 2008-2014, he was the President of the Neurology and Pediatric Neurology Commission at the Romanian Ministry of Health.Among his original scientific contributions, as part his doctoral thesis defended in 1993, he introduced the concept of incipient cognitive impairment for the first time in Romania, referring to patients with cerebrovascular and coronary diseases who develop neurocognitive disorders as a condition preceding the onset of dementia. He has also promoted the implementation of systematic neurocognitive evaluation for patients with neurological and vascular diseases, as well as their treatment in the current pursuit/activity of the neurologists in Romania. Dr. B\u0103jenaru conducted the first brain connectomics study in the context of a neurological condition (Parkinson\u2019s Disease) in Romania. Under the coordination of Professor B\u0103jenaru, a modern school of clinicians and researchers in neurosciences was born, forming several professors and heads of clinics and departments in Bucharest. Throughout his career, he has received numerous awards that certify the effort made and the unquestionable dedication which he had in helping those in need. Among the awards and distinctions received we mention the Romanian Patriarchate Award \u201c100 years of Modern Medicine in the capital of united Romania\u201d (2018), The Excellence Award for the entire activity in the field of Neurology in Romania of the Academica Foundation (2016), the Excellence Award for the whole of the activity of the Foundation Prof. Dr. Marin Voiculescu \u201c(2015), Excellence Award of the International Brain Foundation and the Romanian Academy of Medical Sciences - for his activity in the field of neurology in Romania, (2014); Doctor Honoris Causa of the \u201cOvidius\u201d University of Constan\u021ba, Romania (2006); Award of Excellence in Neurology in Romania - for scientific and educational activity at national level, 2005, 2006 and 2011; Romanian Society of Internal Medicine Award for the most active scientific collaborator in a related specialty, 2008, 2012; Excellency prize of the International Society for the Study of Neuroprotection and Neuroplasticity, 2011; Excellence Award for medical activity awarded by the Romanian Ministry of Health in 2015.He was an influencing neurologist, a fantastic colleague, and a dear friend. It is a great loss for all of us that he has passed away at such a young age. Although he is no longer with us, his powerful impact on our field will last for a very long time. Our deepest sympathy goes to his family in this time of grief.He will be greatly missed. May his soul rest in peace."} +{"text": "The National Institute on Aging (NIA) at the National Institutes of Health, Department of Health and Human Services, is the federally designated lead agency on aging research, and has supported significant research on aging as a life-long process. In the last five years, NIA experienced a tripling of its budget. Although much of this funding is targeted to Alzheimer\u2019s disease (AD) and AD related dementias (ADRD) research, there was an increase in funds allocated to non-AD research in keeping with the overall growth of NIH. This symposium will provide a forum for exploration of the implications of the budget increases for the general research community. It will involve NIA\u2019s senior staff discussing research priorities and programs supported by the Institute. A question-and-answer session will follow brief introductory remarks on current funding and future priorities and research directions of NIA."} +{"text": "The fourth speaker is Dr. Erica Solway. Dr. Solway will discuss her experience working in policy at the federal level as a Health and Aging Policy Fellow and a policy advisor with the U.S. Senate Committee on Health, Education, Labor, and Pensions Subcommittee on Primary Health and Aging and in her current involvement in policy-relevant research at the state and national level at the University of Michigan Institute for Healthcare Policy & Innovation. Dr. Solway is a senior project manager forthe evaluation of the Healthy Michigan Plan, Michigan\u2019s Medicaid expansion program, and is also an associate director of the University of Michigan National Poll on Healthy Aging."} +{"text": "The authors wish to add the following statement in the Acknowledgements section of this paper :The authors would like to acknowledge Dr. Nat\u00e1lia Tobar, Division of Nuclear Medicine, Department of Radiology, School of Medical Sciences, University of Campinas, for the excellent acquisition and discussion of animal parameters by DEXA.The authors apologize for any inconvenience caused and state that the scientific conclusions are unaffected."} +{"text": "The session will also include the presentation of the2020 Lawton Award to recipient Sara J. Czaja, PhD, FGSA. The M. Powell Lawton Award is presented annually to an individual who has made outstanding contributions from applied research that has benefited older people and their care. The Lawton Award is generously funded by the Polisher Research Institute of the Madlyn and Leonard Abramson Center for Jewish Life.\u201d"} +{"text": "Health Resources and Service Administrator-HRSA, grant # K01HP33459 to Dr. Tabrizi. These statements should also be added per funders request.In the original article, we neglected to include the funder \u201cThis publication was made possible by Grant Number K01HP33459 from the Health Resources and Services Administration (HRSA), an operating division of the U.S. Department of Health and Human Services. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the Health Resources and Services Administration or the U.S. Department of Health and Human Services.\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The funding statement for this article should read as follows:\u201cThe research was supported by grants from the European Research Council , the Israel Science Foundation to MS (300/17) and through its Center of Excellence in Structural Cell Biology to SJF and DF (1775/12), a research grant from Sheri and David E. Stone and by a charitable donation from Sam Switzer and family. M.S. is an incumbent of the Aharon and Ephraim Katzir Memorial Professorial Chair. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "There are errors in the Funding statement. The correct Funding statement is as follows: This research/work was supported by the Cluster of Excellence Cognitive Interaction Technology 'CITEC' (EXC 277) at Bielefeld University to SF and AM, which is funded by the German Research Foundation (DFG)."} +{"text": "Correction to:Perspect Med Educ 201910.1007/s40037-019-00558-zUnfortunately information regarding Paul Worley\u2019s affiliation is missing from the original article. Please find the information here:Paul Worley is affiliated to the Prideaux Centre for Research in Health Professions Education, Flinders University, Adelaide, Australia. He is the Australian National Rural Health Commissioner. This is an independent statutory position. The views in this paper are those of the authors only and do not represent the views of the Australian Government or the Department of Health."} +{"text": "There is an error in the Funding statement. The correct number for the National Natural Science Foundation of China is No. 81771231, 81471156 to HJ and No. 81600995 to YS.the National Natural Science Foundation of China, No. 81974176 to HJ, 81901169 to ZC, No. 81901305 to CW.We also neglected to include the funder The final funding part is as follows:This study was supported by the National Key Research and Development Program of China (Nos. 2016YFC0901504 and 2016YFC0905100 to HJ and No. 2016YFC1306000 to BT), the National Natural Science Foundation of China , the Key Research and Development Program of Hunan Province (No. 2018SK2092 to HJ), the Scientific Research Foundation of Health Commission of Hunan Province (No. B2019183 to HJ), and the Clinical and Rehabilitation Fund of Peking University Weiming Biotech Group (No. xywm2015I10 to HJ).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "In the published article, there was an error regarding the affiliations for Sean M. Bialosuknia. Instead having affiliations 1,2,3, they should have affiliations 1,3.In the original article, we neglected to include funding from Pacific Southwest Regional Center of Excellence for Vector-Borne Diseases funded by the U.S. Centers for Disease Control and Prevention (Cooperative Agreement 1U01CK000516) to Erin Taylor Kelly and Geoffrey M. Attardo.This publication was supported by the Cooperative Agreement Number U01CK000509 funded by the Center for Disease Control and Prevention. Its content is solely the responsibility of the authors and do not necessarily represent the official views of the Center for Disease Control and Prevention or the Department of Health and Human Services. The study received funding from Pacific Southwest Regional Center of Excellence for Vector-Borne Diseases funded by the U.S. Centers for Disease Control and Prevention (Cooperative Agreement 1U01CK000516), to EK and GA. We would like to extend our gratitude to Illia Rochlin of Suffolk County Health Department for providing us with Ae. albopictus mosquitoes and the Arbovirus laboratory insectary crew for mosquito maintenance. Data for these analyses were acquired at the University of California, Davis West Coast Metabolomics Center, U2C ES030158. We would like to express our gratitude to Dr. Alice Trimmer for her suggestions and edits.The authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The following information is missing from the Funding statement: This study was funded in part by the Deputyship for Research and Innovation, \u2018Ministry of Education\u2019 in Saudi Arabia through the Project no. IFKSURP-160. No additional external funding was received for this study.The following information is missing from the Acknowledments section: The author extends their appreciation to the Deputyship for Research and Innovation, \u2018Ministry of Education\u2019 in Saudi Arabia for funding this research work through the Project no. IFKSURP-160. Special thanks for Dr Emad Mahmoud, an assistant professor, and senior researcher at King Saud University, for the support in evaluating the eligibility of the articles for inclusion in this systematic review and meta-analysis."} +{"text": "Since 2015, The John A. Hartford Foundation has been funding strategies to create Age-Friendly Health Systems (AFHS). Led by the Institute of Healthcare Improvement, in partnership with the American Hospital Association and the Catholic Health Association, the AFHS movement is rapidly growing, with participation in all 50 states from over 450 sites, including the full continuum of care settings. Partnerships with private and public entities are accelerating the work. As one example, the Health Resources and Services Administration has embedded AFHS principles into the Geriatrics Workforce Enhancement Program. This Kent Lecture will focus on the genesis and trajectory of the AFHS social movement and discuss how the effort will lead to an age-friendly ecosystem that transcends boundaries and cultures and leads to a common framework for the way we approach care, caregiving and communities for optimizing the lives and wellbeing of all older adults."} +{"text": "In the original article, we neglected to include the funder, the Jacobs Foundation, Grant: 015 11 63 00 to Kate Ellis-Davies.The updated Funding section reads as follows:This research was supported, under the auspices of the Open Research Area , by grants from the UK Economic and Social Research Council , The Netherlands Organisation for Scientific Research , the French Agence Nationale de la Recherche , and the Jacobs Foundation (Grant 015 11 63 00 to KE-D) whose support is gratefully acknowledged.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "There are errors in the Funding statement. The publisher apologizes for these errors. The correct Funding statement is as follows: This Collection was supported by TEPHINET, a program of The Task Force for Global Health, Inc., via Cooperative Agreement number NU2GGH001873, funded by the Centers for Disease Control and Prevention. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the Centers for Disease Control and Prevention, the U.S. Department of Health and Human Services, The Task Force for Global Health, Inc., or TEPHINET."} +{"text": "Cross-border partner interactions were supported by the EU Twinning project VACTRAIN #692293 and PI project of the Swedish Institute #19806_2016. Participation of Petkov S, Chiodi F, and Wahren B was supported by VACTRAIN. Participation of Svirskis S was supported by BALTINFECT.There are errors in the Funding statement. The grant number listed from the Russian Foundation of Basic Research is actually from the Russian Science Foundation. The correct Funding statement is as follows: The study was supported by the Russian Foundation of Basic Research project #17_04_00583 Click here for additional data file."} +{"text": "European Immunization Week (EIW) 2020, the European Vaccination Information Portal was launched on 21 April. This publically accessible website provides anyone interested in vaccination with accurate, objective and up-to-date evidence on vaccines and vaccination with an European Union (EU) perspective.On the occasion of Council Recommendation on Strengthened Cooperation against Vaccine Preventable Diseases adopted in December 2018. The portal, which is available in all of the EU\u2019s official languages, features an overview of the existing mechanisms in the EU to ensure that available vaccines conform to the highest standards of safety and effectiveness. It also highlights the benefits of vaccination to the individual and the community. The project is an initiative of the EU, following the The website was developed by the European Centre for Disease Prevention and Control (ECDC), in partnership with the European Commission (EC), specifically, the Directorate-General for Health and Food Safety (DG SANTE) and the European Medicines Agency (EMA).here.EIW, which is led and coordinated by the World Health Organization Regional Office for Europe (WHO/Europe), is marked across the WHO European Region every April. It aims to raise awareness of the importance of immunisation for people\u2019s health and well-being. ECDC supports the EIW campaign by providing scientific evidence on immunisation. Information on ECDC activities in the context of EIW can be found"} +{"text": "The New York City Department of Health and Mental Hygiene, Bureau of Alcohol and Drug Use Prevention, Care and Treatment affiliation should not have been included for Ms Warren. In addition, Dr Kolodny should have been named as the corresponding author instead of Ms Warren. This article has been corrected.1In the Research Letter titled \u201cTrends in Heroin Treatment Admissions in the United States by Race, Sex, and Age,\u201d"} +{"text": "Dr. Takashi Sugimura, an honorary member of the Japanese Environmental Mutagen Society (JEMS), passed away on September 6, 2020, at the age of 94. Dr. Sugimura was a leading international figure in cancer medicine for many years, and was a pioneer in molecular biology. He graduated from the University of Tokyo, Faculty of Medicine, in 1949, received his Ph.D. in 1957, and went abroad to study at the National Cancer Institute in the United States. He then joined the National Cancer Center Research Institute in Tokyo in 1962 and was appointed Director in 1974. He later concurrently served as a Professor of Molecular Oncology at the Institute of Medical Science in the University of Tokyo from 1970 to 1988. He was appointed to be President of the National Cancer Center in 1984 and was instrumental in promoting the government\u2019s Comprehensive 10-Year Strategy for Cancer, which was launched that year, and was appointed Honorary President of the National Cancer Center in 1992. He also served as President of Toho University from 1994 to 2000, and was named President Emeritus in 2000. In 2003, he was diagnosed with stomach cancer and had his entire stomach removed. In an effort to help other patients and their families, he wrote a book about the testing and treatment process and his life after surgery, and continued to work on the development of cancer treatments from the patient\u2019s perspective.Dr. Sugimura\u2019s research clarified the relationship between mutagenicity and carcinogenicity and produced major contributions to the scientific study of cancer development mechanisms. He is especially remembered for his work on the now-well-known fact that cancer is a disease caused by genetic mutations. Together with Dr. Minako Nagao (who later became Chief of the Carcinogenesis Division) and other researchers, he showed that common cooking practices can produce mutagens exhibiting the same structures as those of heterocyclic amines in food. He then isolated and identified a large number of these heterocyclic amines, and showed that they are taken into the body via food consumption and induce mutations in genes, resulting in the development of tumors. He also analyzed multi-step carcinogenesis at the molecular level and studied its relevance to potential avenues for the prevention of cancer. This research has significantly impacted the identification of new mechanisms of chemical carcinogenesis and tumor promotion.Dr. Sugimura was the second President of JEMS for 4\u2009years from 1978 and was the chairman of 5th meeting of JEMS hosted in 1976 in Tokyo. In 1981, he was a President for Third International Conference of Environmental Mutagens, which was held in Tokyo Japan and hosted by JEMS on behalf of International Association of Environmental Mutagen Societies . He also became an honorary member of JEMS in 1987, and was a frequent lecturer at annual JEMS meetings. In addition, in 1994, when the JEMS Award was established, the first prize was awarded for his \u201cResearch on the mutagenicity and carcinogenicity of heterocyclic amines.\u201dThe International Symposium of the Princess Takamatsu Cancer Research Fund, held in Tokyo every autumn, introduces the in latest cancer research and provides a forum for interaction between foreign and domestic researchers. Dr. Sugimura served as the Chair of the Scientific Advisory Committee for this fund for 25\u2009years, from 1989 to 2014, and made significant contributions to the Fund\u2019s activities, including playing a key role in establishing the AACR Princess Takamatsu Memorial Lecture Prize in 2007. He was also instrumental in the establishment of the successful Joint Conference of the American Association for Cancer Research and the Japanese Cancer Association, which began in 1989, and he co-chaired the first meeting with the former President of the AACR, Dr. Enrico Mihich. This joint conference is held every 3\u2009years in Hawaii, and continues to symbolize the close relationship between cancer science researchers in Japan and the United States.Dr. Sugimura\u2019s work has been recognized in the United States as well in Japan, appearing five times on the cover of Cancer Research. He has also been the recipient of multiple honors in the United States, with his election as a member of the AACR in 1969, an honorary member in 1980, and the first fellow of the AACR Academy in 2013. His accomplishments are too numerous to list in full here, so I will only list some of the major awards he has received as evidence of his high esteem: the Imperial Prize and Japan Academy Prize in 1976, the EMS Award from the Environmental Mutagen Society in 1978, the Order of Culture of the Japanese Government in 1978, the Charles S. Mott Prize from the General Motors Cancer Research Foundation in 1981, the Tomizo Yoshida Prize from the Japanese Cancer Association in 1992, and the Japan Prize for Biotechnology in Medicine in 1997. He also served as the 25th President of the Japan Academy in 2013\u201316. He was an elected foreign associate of the National Academy of Sciences and the Institute of Medicine (U.S.), and an elected foreign member of the Royal Netherlands Academy of Arts and Sciences and the Royal Swedish Academy of Sciences.There is an expression in Japanese, \u201cfall of a giant star.\u201d In the field of genotoxicity and cancer research, Dr. Sugimura\u2019s death may be best described using this expression. It is an undeniable fact that Dr. Sugimura was and will continue to be respected for his intellectual contributions to research, his amazing leadership skills, and his supreme passion for research above all.May he rest in peace.Editor-in-ChiefMasami YamadaNational Defense Academy of Japan"} +{"text": "In the published article, there was an error regarding the affiliation for Lidan Sun. Instead of affiliation 3, it should be 2.In addition, there is an error in the order of the Funding statement. The correct statement appears below. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.This research is supported by the National Natural Science Foundation of China (No. 31870689), the Fundamental Research Funds for the Central Universities (NO. 2015ZCQ-SW-06), the National Natural Science Foundation for Young Scientists of China , and grant 201404102 from the State Administration of Forestry of China."} +{"text": "The authors wish to make the following erratum to this paper . The FunThis research was funded by project (RTI2018-093321-B-I00) supported by: ERDF , Ministry of Science and Innovation and State Research Agency from Spain.Life editorial office has been asked to update the original manuscript [The authors would like to apologize for any inconvenience caused to the readers by these changes. The nuscript on the a"} +{"text": "There are errors in the Funding statement. The correct Funding statement is as follows: This Collection was supported by TEPHINET, a program of The Task Force for Global Health, Inc., via Cooperative Agreement number NU2GGH001873, funded by the Centers for Disease Control and Prevention. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the Centers for Disease Control and Prevention, the U.S. Department of Health and Human Services, The Task Force for Global Health, Inc., or TEPHINET."} +{"text": "The past 5 years have seen incredible advances in approaching hearing loss as a major public health issue. National efforts include the 2015 President\u2019 Council of Advisors on Science and Technology and the National Academies of Science, Engineering, & Medicine\u2019s 2016 Commission on Hearing Health Care for Adults, which led to the 2017 OTC hearing aid legislation and the expected debut of OTC hearing aids in 2020-2021. The World Report on Hearing amplifies these efforts. This presentation will cover the role of the Report in the context of the rapidly evolving hearing care landscape in the US and how the Report\u2019s call for affordable, accessible hearing care fit within current national efforts focused on older adults. Finally, the WHO recognized 2020-2030 as the Decade of Healthy Aging. We will discuss how the World Report on Hearing integrates with broader efforts to support healthy aging locally and globally."} +{"text": "The Clark Tibbitts Award lecture will feature an address by the 2020 award recipient, Jan Abusharkrah, PhD, FAGHE. AGHE\u2019s Clark Tibbitts Award was established in 1980 and named for an architect of the field of gerontological education. The award is given each year to an individual or organization that has made an outstanding contribution to the advancement of gerontology and geriatrics education."} +{"text": "The manuscript draft was developed by KL, MG, and MKG. All authors contributed to the article and approved the submitted version.YL is supported by National Key Research and Development Program of China (2017YFC1310405) and the National Natural Science Foundation of China (U1736124). MKG is supported by the Bekker programme grant of the Polish National Agency for Academic Exchange (PPN/BEK/2019/1/00245/U/00001).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "In 2015, the Program of Merit was expanded and adapted to implement a voluntary evaluation process for health professions programs that are choosing to integrate gerontology/geriatrics competencies in order to prepare students for working with older adults as well as their informal care partners. These programs are now eligible to apply for the Program of Merit designation. The Program of Merit for Health Professions Programs is based on the AGHE Standards and Guidelines for Gerontology/ Geriatrics in Higher Education, Sixth Edition (2015), specifically Chapters 11 and 12. This session will provide an overview of the process, review the application content, and provide technical support for interested participants."} +{"text": "Correction to: BMC Pregnancy and Childbirth (2020) 20:257https://doi.org/10.1186/s12884-020-02951-7Following publication of the original article , the autAcknowledgementsWe would like to thank the team of the Netherlands\u2019 Course on Global Health and Tropical Medicine of the KIT Royal Tropical Institute in Amsterdam for passionately teaching the determinants of health model. The lectures and assignments of the course led to the first draft of this paper. Especially the guidance and support of Maaike Flinkenfl\u00f6gel was of great value. We thank professor Florence Mirembe from the Department of Obstetrics and Gynaecology of the Makerere University College of Health Sciences in Kampala, Uganda, for sharing her thoughts after reading our manuscript."} +{"text": "There is information missing from the Funding statement. The complete, correct Funding statement is as follows: This research was made possible in part by a grant from NOAA Fisheries, Office of Protected Resources, Species of Concern Program, in part by a grant from The Gulf of Mexico Research Initiative, and in part by funding from the Florida Fish and Wildlife Conservation Commission (Grants FWC-08304 and FWC-16188 to WFP). Riverside Technology, Inc. provided support in the form of salary for author LT, but did not have any role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of this author are articulated in the 'author contributions' section.The publisher apologizes for the error.https://data.gulfresearchinitiative.org (doi:10.7266/N7W37TWR).The following information is missing from the Data Availability statement: Dissolved inorganic carbon 14C data are publicly available through the Gulf of Mexico Research Initiative Information & Data Cooperative (GRIIDC) at"} +{"text": "The National Institute on Aging (NIA) at the National Institutes of Health, Department of Health and Human Services, is the federally designated lead agency on aging research, and has supported significant research on aging as a life-long process. In the last five years, NIA experienced a tripling of its budget. Although much of this funding is targeted to Alzheimer\u2019s disease (AD) and AD related dementias (ADRD) research, there was an increase in funds allocated to non-AD research in keeping with the overall growth of NIH. This symposium will provide a forum for exploration of the implications of the budget increases for the general research community. It will involve NIA\u2019s senior staff discussing research priorities and programs supported by the Institute. A question-and-answer session will follow brief introductory remarks on current funding and future priorities and research directions of NIA."} +{"text": "The authors of \u201cPostvaccination Fever Response Rates in Children Derived Using the Fever Coach Mobile App: A Retrospective Observational Study\u201d :e12223) identified an error in the Acknowledgements section.In the first part of the Acknowledgements section, the following phrase:This work was supported by Technology Innovation Program (10060085) funded by the Ministry of Trade, Industry, and Energy Has been changed to:This work was supported by Technology Innovation Program (20002289) funded by the Ministry of Trade, Industry, and Energy The correction will appear in the online version of the paper on the JMIR website on May 7, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "The following information is missing from the Funding statement: This paper is an output of a research project implemented as part of the Basic Research Program at the National Research University Higher School of Economics (NRU HSE).There is an error in affiliation 1 for authors Iuliia Naidenova, Petr Parshakov, and Aleksei Chusovliankin. The correct affiliation 1 is: HSE University, Perm, Russia."} +{"text": "The authors wish to make the following corrections to this paper . The authors would like to acknowledge Dr. Ali Khammanivong for providing gRNA sequences and guidance during the generation of the CD44 knockout cell lines. The authors would also like to acknowledge the Burroughs Wellcome Fund and the Howard Hughes Medical Institute Medical Research Fellowship program for providing funding for Dr. Witschen.The original \u201cAcknowledgments\u201d isAcknowledgments: The authors would like to thank Douglas Yee (funded by Masonic Cancer Center (MCC) Support Grant, P30-CA077598), and other members of the University of Minnesota Breast Cancer Translational Working Group (BrCa-TWG), for assistance with the cell line and patient Nanostring gene expression experiments. The authors would also like to thank the Elsa Pardee foundation .And should be replaced with:Acknowledgments: The authors would like to thank Douglas Yee (funded by the Masonic Cancer Center (MCC) Support Grant, P30-CA077598), and other members of the University of Minnesota Breast Cancer Translational Working Group (BrCa-TWG), for assistance with the cell line and patient Nanostring gene expression experiments. The authors would also like to thank the Elsa Pardee foundation . The authors would like to acknowledge Ali Khammanivong for providing gRNA sequences and guidance during the generation of the CD44 knockout cell lines. The authors would also like to acknowledge the Burroughs Wellcome Fund and the Howard Hughes Medical Institute Medical Research Fellowship program for providing funding for Patrice M. Witschen.The authors would like to apologize for these omissions from the original manuscript. These changes do not affect the scientific results. The manuscript will be updated, and the original will remain online on the article webpage."} +{"text": "Gordon has been a mentor for me, and one of my best friends for 40 years. What has always impressed me of Gordon was his innate ability to bring people to work together, to build new structures and to facilitate cancer research in creative new ways. Those who knew him will remember his sharp thinking, his wit and great sense of humor and his upbeat spirit and love for life, work and fun. I will never forget his kindness, his balanced advice, his jokes, and his incredibly fast speaking.Professor Gordon McVie qualified in the nineteen sixties in science and medicine at Edinburgh University, he was appointed Foundation Senior Lecturer at the Cancer Research Campaign oncology unit at the University of Glasgow in 1975. He trained in the US, and spent sabbaticals in Paris, Sydney, and Amsterdam.Throughout the eighties, he was Clinical Research Director at the Netherlands Cancer Institute in Amsterdam, and set up their drug discovery programme and the leading intraperitoneal chemotherapy studies in Europe. In the early eighties he was the Chair of the European Organization for Research and Treatment of Cancer (EORTC) Lung Cancer Cooperative Group for 6 years. I met him for the first time in Brussels at one of the first meetings of this newly formed group. He was also appointed Chair of the UICC Fellowships Programme in 1990 for 8 years. From 1994 to 1997 he served as President of the EORTC, and during that time he set up the present Drug Development Group in Brussels, and with the US National Cancer Institute support, the European New Drug Development Network.After almost a decade in the Netherlands, Professor McVie returned to the UK to be the Chief Executive of the Cancer Research Campaign (CRC), which, under his aegis, took over 70 molecules from the lab into clinical trial. He led CRC into a merger with Imperial Cancer Research Fund in 2002, which formed the largest cancer charity in the UK, Cancer Research UK. In the UK he was one of the architects of the Cancer Trials Networks in Scotland, Wales, and England, and was a founding member of the National Cancer Research Institute. His commitment to drug discovery and delivery is evidenced by ~240 patents granted to CRC scientists under his leadership, several drugs registered including carboplatin, temozolomide, olaparib, and abiraterone and the foundation of 10 biotechnology companies based on CRC intellectual property. His clinical interests, apart from new drug discovery and chemoprevention were in the management of cancers of the lung, ovary, liver, breast, and brain.ecancer.org and ecancerpatient.org both online Open Access free websites, with almost 21 million visits in a decade from 191 countries. For 10 years he was Senior Consultant building clinical research at the European Institute of Oncology in Milan.Lately he was visiting professor at Cancer Studies, Kings College London for five years, and was Clinical Research Advisor to the Institute of Molecular Oncology (IFOM) in Milan. He was founding editor of He was a Director and Co-Founder of Ellipses Pharma - an advanced cancer medicines company.Professor McVie was the recipient of numerous awards and has honorary doctorates in science and medicine from six universities. He has served on key committees of AACR and ASCO, and on the boards of the National Cancer Institutes of France, Italy, and Holland. He has authored 360 peer-reviewed articles, and contributed to over 35 books.He was elected as Fellow of the European Association for Cancer Science in 2014, and has been chairman of the European Alliance for Personalized Medicine since early 2016.This is only a short selection of the most important activities Prof. McVie has been engaged in. He was truly a dynamic international presence that recognized the necessity of going beyond the geographical boundaries, bringing people together to create novel structures in the pursuit of the fight against cancer.The author confirms being the sole contributor of this work and has approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "This presentation will describe collaborative efforts on the campus of a mid-sized, private university to carry out activities consistent with the Age-Friendly University philosophy. In one program, staff from Career Services and a faculty member from the Department of Health Science coordinated with the President\u2019s College (a continuing education program for adult learners), the Emeriti Association (a group of retired faculty members), and alumni to offer mock interviews for students preparing for graduate school. In another program, steps were taken to coordinate with the office of Diversity, Equity and Inclusion to address Ageism in the Workplace. The presentation will conclude with advice for identifying allies across campus and fostering support for the AFU principles."} +{"text": "In the Sample and procedure subsection of the Methods, there is an error in the first sentence of the first paragraph. The correct sentence is: The ESA is regularly conducted by the Institute for Therapy Research (IFT Institut f\u00fcr Therapieforschung).Information is missing from the Acknowledgments. The correct Acknowledgments are as follows: We thank the IFT Institut f\u00fcr Therapieforschung for providing the data for this analysis.There are errors in the Funding statement. The correct Funding statement is as follows: The 2015 Epidemiological Survey of Substance Abuse (ESA) was supported by funding from the German Federal Ministry of Health (project no. IIA5 \u20132514DSM200)."} +{"text": "Scientific Reports 10.1038/s41598-019-54310-y, published online 09 January 2020Correction to: The Acknowledgements section in this Article is incomplete.\u201cThe authors would like to thank CReATE and the ALS Association (US) for the award of the grant supporting the study: \u201cGoing Dry: empowering neurofilament-based biomarkers studies for disease monitoring in amyotrophic lateral sclerosis\u201d. We would also like to thank all ALS patients and their families for their support to this research project and we acknowledge the MND Association (UK) for their help in establishing the ALS biomarkers biobank and contributing with significant resources in the sample collection phase and to general aspect of research in ALS. We also acknowledge the support of the North-Thames Local Research Network (London) in the recruitment process and in the collection and processing of biological samples.\u201dshould read:\u201cThe authors would like to thank CReATe and the ALS Association (US) for the award of the grant supporting the study: \u201cGoing Dry: empowering neurofilament-based biomarkers studies for disease monitoring in amyotrophic lateral sclerosis\u201d. The CReATe consortium (U54NS092091) is part of the Rare Diseases Clinical Research Network (RDCRN), an initiative of the Office of Rare Diseases Research (ORDR), NCATS. CReATe is funded through a collaboration between NCATS, and the NINDS. We would also like to thank all ALS patients and their families for their support to this research project and we acknowledge the MND Association (UK) for their help in establishing the ALS biomarkers biobank and contributing with significant resources in the sample collection phase and to general aspect of research in ALS. We also acknowledge the support of the North-Thames Local Research Network (London) in the recruitment process and in the collection and processing of biological samples.\u201d"} +{"text": "The affiliation for the third author is incorrect. The correct affiliation is not indicated. Christian Raschner is not affiliated with #2 but with: Department of Sport Science, University of Innsbruck, Innsbruck, Austria."} +{"text": "The authors wish to make the following correction to this paper :In the original version of our article , 8897), insufficient source of funding was given. The authors wish to change the information in the Funding section from:Funding: This research was funded by German Academic Exchange Services, grant number 91727334.to the correct version as follows:Funding: This research was funded by the German Academic Exchange Services (DAAD), grant number 91727334, and by the Deutsche Forschungsgemeinschaft \u2014Projektnummer 427397520.The authors would like to apologize for any inconvenience caused to the readers by these changes."} +{"text": "The Donald P. Kent Award lecture will feature an address by the 2019 Kent Award recipient, Terry Fulmer, PhD, of The John A. Hartford Foundation. The Kent Award is given annually to a member of The Gerontological Society of America who best exemplifies the highest standards of professional leadership in gerontology through teaching, service, and interpretation of gerontology to the larger society. The Robert W. Kleemeier Award lecture will feature an address by the 2019 Kleemeier Award recipient, Steven Zarit of Pennsylvania State University. The Kleemeier Award is given annually to a member of The Gerontological Society of America in recognition for outstanding research in the field of gerontology"} +{"text": "In the original article, we neglected to include the funder National Institutes of Health, Eunice Kennedy Shriver National Institute of Child Health and Human Development, P50HD089922, to Giorgia Picci and Emma Rose.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way."} +{"text": "The following information is missing from the Funding statement: This study was funded by a grant to Dr. Ahuja from Astex Inc. (GRANT # 120039).There is information missing from the Competing Interests statement. The correct Competing Interests statement is as follows: Dr. Ahuja receives research grant funding from Astex Inc. and the Van Andel Research Institute. She is a consultant for and has licensed methylation biomarkers to Cepheid (PATENT # 10167513). Dr. Ahuja has also served as consultant to Johnson and Johnson, an advisor to Celgene, and a member of the Scientific Advisory Council to the No Stomach for Cancer Foundation. This does not alter our adherence to PLOS ONE policies on sharing data and materials.The raw data underlying the findings of this study are missing from the list of Supporting Information. The authors have provided the data as Supporting Information file S1 Data(XLSX)Click here for additional data file."} +{"text": "The 2019 marks twenty years since Professor Ao Li (\u9ece\u9ccc) passed away. Although he has left us, the \u201cLi Ao Spirit\u201d has lived on and continuously encourages us to climb higher in scientific research and make more contributions to our people and country , was born in Changsha, Hunan Province in May 1917. He graduated from the National Shanghai Medical College in 1941, and had a long and distinguished career in medicine. His various positions over the years included Associate Professor at the National Chung Cheng Medical College, Professor at the Third Military Medical University, Director of the Burn Research Institute of the Third Military Medical University, and the Vice President of the Third Military Medical University. As one of the pioneers and founders of burn medicine in China, professor Li led comprehensive multidisciplinary research on the pathogenesis and prevention of inhalation injury. For his great contribution to the field of burn research, he was not only the recipient of the Everett Idris Evans Memorial Lecture Award of the American Burn Association in 1994, but also won first prize in both the Chinese Science and Technology Advancement Award and the Military Scientific and Technological Progress Award among other accolades.After graduating from high school, Mr. Li was recommended to study in the Department of Physics or Mathematics at Nanjing Central University. However, touched by the sufferings of the general public caused by a lack of medical knowledge and medicine and his father\u2019s illness, Mr. Li was determined to become a doctor. In 1935, he was given the opportunity to study at Shanghai Medical College, where he started his medical career. After graduating, he successively presided over general surgical work in the National Chung Cheng Medical College, the Third Military Medical University and other institutions, saving thousands of lives.In the late 1950s, the number of burn patients increased drastically due to the country\u2019s efforts in increasing steel production, but there were no effective therapies available at that time , which was published by People\u2019s Military Medical Press in 1993 launched the research project named \u201cStudy on the Pathogenesis of Early Burn Injury and Wound Healing Mechanism\u201d. The project was sponsored by the National Natural Science Foundation. After five years of hard work, they discovered that macrophages act as an initiator in the uncontrolled inflammatory response after burns, verified that vascular endothelium played the central role in the pathogenesis of early organ damage, clarified the mechanism of the early intestinal damage and intestinal infections, and proposed corresponding treatment measures (Huang et al., Professor Li attached great importance to the growth and cultivation of young scientists. He emphasized training in practice, encouraged young scientists to participate in clinical operation, classroom teaching and laboratory tasks Fig.\u00a0. He provIn 1986, Professor Li won the National Science and Technology Progress Award. He donated the bonus of the award to establish the Outstanding Paper Award in Burn Medicine for Young Army Scientists to encourage young scientists and technicians to engage in burn research. In 1996, Professor Li won the China Engineering Science and Technology Award and Military Technical Major Contribution Award. Again, he donated the bonus of both awards, a total of 150,000 Chinese Yuan, to set up the Li Ao Burn Medicine Fund so as to reward young and middle-aged scientists who contributed to the development of burn medicine. Ten of the recipients of the Li Ao Burn Medicine Fund have become the leading scientists of burn medicine in China. In 1996, Professor Li was awarded the title of \u201cOutstanding Mentor\u201d by the General Logistics Department of Chinese People\u2019s Liberation Army.Professor Li has devoted his whole life to the cause of burn medicine in China and made a lot of remarkable achievements. Although he left us twenty years ago, we are still always encouraged and inspired by the \u201cLi Ao Spirit\u201d of selflessness, diligence and entrepreneurship. With the great efforts of generations of doctors and scientists, we believe burn medicine research in China will bring about more and more breakthroughs and play a leading role in the international arena."} +{"text": "UNICEF is acknowledged for their support towards the Center for Nutrition Studies, Yenepoya (Deemed to be University). No additional external funding was received for this study.There are errors in the Funding statement. The correct Funding statement is as follows: MB received a grant from The United Nations Children's Fund (UNICEF) for a capacity building workshop on Nutrition Assessment Techniques. The URL of the website is:"} +{"text": "There is an error in the Funding statement. The correct number for the National Natural Science Foundation of China is No. 81771231 to HJ and No. 81600995 to YS.the National Natural Science Foundation of China, No. 81974176 to HJ; No. 81901169 to ZC. The corrected funding statement appears below.Additionally, we neglected to include the funder We declared that there were no financial interests in this study. This study was supported by the National Key Research and Development Program of China , the National Natural Science Foundation of China , the National Natural Science Foundation of Hunan Province (No. 2019JJ40363 to RQ), Key Research and Development Program of Hunan Province (No. 2018SK2092 to HJ), Scientific Research Foundation of Health Commission of Hunan Province (No. B2019183 to HJ), The Clinical and rehabilitation fund of Peking University Weiming Biotech Group (No. xywm2015I10 to HJ).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Bioengineering and Translational Medicine, we are pleased to introduce our Editorial Board Member, Professor W. Mark Saltzman. Professor Saltzman is the Goizueta Foundation Professor of Biomedical Engineering, Chemical and Environmental Engineering, and Physiology at Yale University. Professor Saltzman received his BS in chemical engineering from Iowa State University before earning a SM in chemical engineering and a PhD in medical engineering from the Massachusetts Institute of Technology (MIT).In this issue of Professor Saltzman's research has been formative in the fields of controlled drug delivery and tissue engineering. As a trainee in the laboratory of Professor Robert Langer at MIT, Professor Saltzman participated in some of the very earliest work in utilizing polymer scaffolds for cell transplantation with Dr Joseph Vacanti.Professor Saltzman is recognized as a pioneer in quantifying and modeling the transport of macromolecules through synthetic polymers and biological matrices,In addition to his research excellence, Professor Saltzman is passionate about education and has left an indelible mark in this area as well. He has contributed to the emergence of the academic field of biomedical engineering by authoring three original textbooksOver the past 30+ years, Professor Saltzman's contributions as a researcher, educator, and mentor have been immense. He has supervised the research work of over 43 PhD students and 27 postdoctoral researchers who now populate the academic and industry landscape and contribute to his legacy of fundamental discovery and translation in bioengineering. His overall excellence has been recognized via election as Fellow of the American Institute for Medical and Biological Engineering (1997), Fellow of the Biomedical Engineering Society (2010), member of the U.S. National Academy of Medicine (2014), and member of the U.S. National Academy of Engineering (2018). Moreover, Professor Saltzman has set a standard for supportive mentorship that has created a family\u2010like network among his trainees. He is always available for advice and support, even for those who have long ago left his group, no matter how many other demands on his time there may be. On behalf of his current and former trainees, I express my appreciation and gratitude to Mark for sharing his scientific journey with us and helping to shape our own through his insight and guidance."} +{"text": "In the original article, we neglected to include the funder European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme, grant agreement No. 723951 to RO. The corrected Funding section reads as follows:This work was supported by the EPSRC Cambridge NanoDTC, EP/L015978/1 and the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme, Grant Agreement No. 723951 to RO.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The secondary analysis presented here was not externally funded. The funder of the original study had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We acknowledge financial support for the publication costs from the Open Access Publication Fund of the University Medical Center Hamburg-Eppendorf (UKE) and the German Research Foundation (DFG).There are errors in the Funding section. The correct Funding statement is: The original study, of which data were used for analysis, was funded by the German Federal Ministry of Education and Research ("} +{"text": "In the published article, there was an error in the Funding statement. The funding from the National Medical Research Council (NMRC) was mistakenly omitted. The correct Funding statement appears below."} +{"text": "The Belt and Road Initiative (BRI) is a development plan proposed by China that aims to build a new platform for international cooperation and create new drivers of shared development. South Asia is a key area in the Belt and Road Initiative, including eight countries. As the BRI implemented, China\u2019s trade with South Asia has been gradually strengthened. This paper explores the influencing factors of China-South Asia trade under the background of the BRI by using Gravity Model of Trade. The results show that economic growth in China and South Asia, increase of savings rate and improvement of industrialization in South Asia has a significant positive effect on China-South Asia trade. While the development gap between China and South Asia has negative effect on China-South Asia trade. The Belt and Road Initiative (BRI) was first proposed by Chinese President Xi Jinping in 2013 . . lnGDPitlnGDPit is negative and significant at 10% level, which is different from the expected sign. This is due to the restriction of economic development level, China mainly imports labor-intensive products from South Asian countries, because countries with lower economic development level have relatively few laborers, and the eight South Asian countries are not China\u2019s main trading partners. The coefficients of lnGDPct, lnGDPPict, lnFDIict and AR(1) is significant at 1% level with the same expected sign. The economic growth in China, China\u2019s invest, the common development of China-South Asia, the path-dependence such as import products from South Asia will encourage South Asia\u2019s export to China. The coefficient of SRit is positive and significant at 5%level. Therefore, the increase of savings rate of South Asia can expand domestic investment of South Asia countries and bring more opportunities to export goods for South Asia.On China\u2019s imports from South Asia as shown in column (3), most facts have significant effect and the same expected sign. The coefficient of BRIit is positive and significant at 10% level in column (3), while not significant in column (1) and (2), This shows that the impact of the Belt and Road Initiative or establish a free trade zone at present is relied on China much more than on South Asia, which was consistent with the findings of Kong (2018) and Zhang (2019) [The coefficient g (2019) , 26. TheTo sum up, we concluded that, GDP, FDI and AR (1) have significant influence on China-South Asia trade, which means that economic growth, development gap, national savings rate and industrialization level are the main influencing factors of trade between China and South Asia. Among them, economic growth, savings rate and industrialization level of both sides have a significant positive impact, while the development gap between China and South Asia has a negative impact on trade between China and South Asia. The impact of BRI on China-South Asia trade is mainly reflected in China\u2019s imports, which shows that the construction of the Belt and Road Initiative still needs to be deepened step by step.The trade volume between China and South Asia has been increasing gradually. But the trade imbalance is obvious\u2014South Asia seriously depend on China\u2019s imports. China mainly exports capital-intensive products to South Asia, while China imports labor-intensive products from South Asia. The economic growth, the development gap, national savings rate and the industrialization level have influence on the trade between China and South Asia. Among these influencing factors, economic growth of China and South Asia counties, savings rate and industrialization level of host countries have significant positive effect on the bilateral trade, while development gap has negative effect. However, the effect of BRI on China-South Asia trade is not significant. The following policy implications are proposed.Promote China\u2019s Going Out Strategy and encourage foreign direct investment. On the one hand, as China improve the value chain and increase the environmental awareness, many capital-intensive and labor-intensive industries, such as construction, manufacturing and energy, are shifting production abroad. Many Chinese multinationals bring advanced technology to invest in countries with rich resources and lower labor costs. On the other hand, expanding China\u2019s investment in South Asia will facilitate technology transfer to domestic firms through spillover, which also promote domestic firms to make the strategic decision to adopt the advanced technology and stimulate innovation and creativity.China and South Asia should deepen the cooperation of production. China has a relatively high level of industrialization. Deepening production capacity cooperation through building production bases and economic and trade cooperation zones can promote the industrialization of South Asia. The improvement of the industrialization expands the range of commodities of China-South Asia trade and optimize the trade structure.Through combing the existing literature, although there are innovations, there are still limitations. For example, in terms of choosing influencing factors, although the research pursues the comprehensiveness of the theme, there may be other factors that have a certain impact on bilateral trade. In addition, the implementation of the Belt and Road Initiative policy is affected by many parties, and the implementation of specific projects and their impact on trade and economy need longer time and data to further prove and analyze. This study suggests that future research should continue to track and explore the trade influencing factors of South Asia and other countries and regions along the Belt and Road Initiative.S1 Data(XLSX)Click here for additional data file."} +{"text": "In January, 2023, the Science and Security Board of the Bulletin of the Atomic Scientists moved the hands of the Doomsday Clock forward to 90\u00a0s before midnight, reflecting the growing risk of nuclear war.Current nuclear arms control and non\u2010proliferation efforts are inadequate to protect the world's population against the threat of nuclear war by design, error, or miscalculation. The Treaty on the Non\u2010Proliferation of Nuclear Weapons (NPT) commits each of the 190 participating nations\u201dto pursue negotiations in good faith on effective measures relating to cessation of the nuclear arms race at an early date and to nuclear disarmament, and on a treaty on general and complete disarmament under strict and effective international control\u201d.Any use of nuclear weapons would be catastrophic for humanity. Even a \u201climited\u201d nuclear war involving only 250 of the 13,000 nuclear weapons in the world could kill 120 million people outright and cause global climate disruption leading to a nuclear famine, putting 2 billion people at risk.http://www.ippnw.org).The health community has had a crucial role in efforts to reduce the risk of nuclear war and must continue to do so in the future.In 2007, the IPPNW launched the International Campaign to Abolish Nuclear Weapons, which grew into a global civil society campaign with hundreds of partner organisations. A pathway to nuclear abolition was created with the adoption of the Treaty on the Prohibition of Nuclear Weapons in 2017, for which the International Campaign to Abolish Nuclear Weapons was awarded the 2017 Nobel Peace Prize. International medical organisations, including the International Committee of the Red Cross, the IPPNW, the World Medical Association, the World Federation of Public Health Associations, and the International Council of Nurses, had key roles in the process leading up to the negotiations, and in the negotiations themselves, presenting the scientific evidence about the catastrophic health and environmental consequences of nuclear weapons and nuclear war. They continued this important collaboration during the First Meeting of the States Parties to the Treaty on the Prohibition of Nuclear Weapons, which currently has 92 signatories, including 68 member states.We now call on health professional associations to inform their members worldwide about the threat to human survival and to join with the IPPNW to support efforts to reduce the near\u2010term risks of nuclear war, including three immediate steps on the part of nuclear\u2010armed states and their allies: first, adopt a no first use policy;The danger is great and growing. The nuclear armed states must eliminate their nuclear arsenals before they eliminate us. The health community played a decisive part during the Cold War and more recently in the development of the Treaty on the Prohibition of Nuclear Weapons. We must take up this challenge again as an urgent priority, working with renewed energy to reduce the risks of nuclear war and to eliminate nuclear weapons.Respective authors were paid by their employers. Chris Zielinski's time was funded by International Physicians for the Prevention of Nuclear War.KB\u2010D is a full\u2010time employee of the American Medical Association, working as the Editor\u2010in\u2010Chief of JAMA and the JAMA Network. AH is principal investigator of the Pathfinder Initiative 2020\u20132025, co\u2010investigator of Sustainable Healthy Food Systems research programme 2017\u20132023, and co\u2010investigator of Complex Urban Systems for Sustainability and Health 2017\u20132023, all funded by the Wellcome Trust, with additional funding from the Oak Foundation for the Pathfinder Initiative, and he reports royalties from Cambridge University Press for the coauthored book Planetary Health; consultancy fees paid to his institution from the Wellcome Trust for his role as Senior Advisor on Climate Change and Health in 2021; travel/meeting support from WHO and Human Frontiers Science Program; and he is a member of the Cool Roofs trial steering committee Nouna Research Centre, Burkina Faso/University of Heidelberg, is Co\u2010chair of the International Advisory Committee, NIHR Clean\u2010Air (Africa) Global Health Research Unit, is a member of the Independent Advisory Group, Collaboration for the Establishment of an African Population Cohort Consortium, and he was Co\u2010chair of the InterAcademy Partnership, Climate Change and Health Working Group 2019\u20132022 and Co\u2010chair of the Academy of Medical Sciences/Royal Society working group on \u2018A healthy future\u2014tackling climate change mitigation and human health together\u2019 2020\u20132021 . IH reports honoraria for several speaking engagements, all donated to Back from the Brink, the International Physicians for the Prevention of Nuclear War, or Physicians for Social Responsibility; travel/ meeting support for Nobel Peace Laureates' Summit, the World Federation of Public Health Associations World Congress, and the UN Human Rights Commission Youth Summit; and he is a member of the steering committee of Back from the Brink and the International Steering Group of the International Campaign to Abolish Nuclear Weapons, a Board member of the International Physicians for the Prevention of Nuclear War and Physicians for Social Responsibility, and a Trustee of the Phillips Exeter Academy . MGMOR reports research grants from the Dutch Research Council, NOW (grant number COMPL.21COV.001) and from the Netherlands Organisation for Health Research ZonMw (grant number 09120012010063) and he is Chair of the Dutch guideline committee on cognitive impairments and dementia. TR reports a contract with the Institute for Energy and Environmental Research (USA) for papers addressing the health and environmental consequences of nuclear testing in multiple locations, including Australia, French Polynesia, central Pacific, and China; honorarium from The Choisun Ilbo media group in South Korea for a lecture on nuclear weapons in 2022 and for nuclear weapons presentations from Hyogo Medical Practitioners Association (Japan), Peace Boat (Japan), and the University of Sydney; he was an expert witness on radiation and health for Environmental Justice Australia acting for Mine\u2010Free Glenaladale regarding proposed Fingerboards Mineral Sands Mine to the Victorian Government Fingerboards Inquiry and Advisory Committee; he is a member of RV3 Rotavirus Vaccine Development Scientific Advisory Board, Murdoch Children's Research Institute/ Royal Children's Hospital; he is a member of the Committee of International Campaign to Abolish Nuclear Weapons Australia; he is a member of the Internet Peace Prize Award Committee ; he was a member of the Victorian International Humanitarian Law Advisory Committee, Australian Red Cross; he is a Board member of the Initiative for Peacebuilding, Faculty of Arts, University of Melbourne; he is an At\u2010large Board member of the International Physicians for the Prevention of Nuclear War; he was Co\u2010president of the International Physicians for the Prevention of Nuclear War 2012\u201323; and he is Honorary Principal Fellow, Melbourne School of Population and Global Health, University of Melbourne. PY reports grants from Atea Pharmaceuticals; honoraria for lectures, presentations, and educational events from bioM\u00e9rieux and Pfizer Pharmaceuticals; fees for participation on an advisory board from Pfizer Pharmaceuticals; and he is a member of the Antimicrobial Stewardship Study Group Executive Committee (2022\u20132024) and the Clinical Practice Guideline Panel on Vaccinations in Immunocompromised hosts for the European Society of Clinical Microbiology and Infectious Diseases. CZ reports consulting fees for his role as senior adviser on the international journals project from the International Physicians for the Prevention of Nuclear War. All the other authors declare no competing interests."} +{"text": "Funding statement.In the published article, there was a mistake in the Original text: This research was supported by the National Alliance for Research on Schizophrenia & Depressio (NARSAD) Young Investigator Award and K23 MH067705 to EN and K01 DA020485 to JE. AK was supported by grant Ministry of Education and Science of the RK AP08856595. JA was supported by R01-DA022221-02S2 (PI: DelBello).Due to an oversight by the authors during proofing, the order of grant support was listed incorrectly, and some awkward wording was not corrected. As a condition of supporting the research publication costs, the Ministry of Education and Science in the Republic of Kazakhstan requires that it must be listed first in the funding statement. The correct Funding statement appears below."} +{"text": "In the published article, there was an error in the Funding statement. The grant number was mistakenly omitted. The correct Funding statement appears below.This material is based upon work supported by, or in part by, the Army Research Laboratory and the Army Research Office under contract/grant number W911NF-19-0419.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The corrected Acknowledgements statement appears below.In the published article, Vounba P, Loul S, Tamadea LF, Siawaya JFD. Microbiology laboratories involved in disease and antimicrobial resistance surveillance: Strengths and challenges of the central African states. Afr J Lab Med. 2022;11(1), a1570. The original incorrect wording:We sincerely thank Dr Skander Hathroubi for his contribution by reading and correcting the first draft of the manuscript.The revised and updated wording:We sincerely thank Dr Skander Hathroubi for his contribution by reading and correcting the first draft of the manuscript. This study was commissioned and supported by the Regional Disease Surveillance Systems Enhancement (REDISSE) project in Central Africa, which is financially supported by the World Bank and managed by the Economic Community of Central African States (ECCAS). We would like to thank ECCAS and the World Bank for supporting our efforts through REDISSE.The authors apologise for this error. The correction does not change the study\u2019s findings of significance or overall interpretation of the study\u2019s results or the scientific conclusions of the article in any way."} +{"text": "Funding statement appears below:In the original article, there was an error in the Funding statement. . The correct \u201cThis study was supported by the project of the National Key R&D Program of China (No. 2019YFC1712105), and the Major Basic Research Project of Shandong Natural Science Foundation (No. ZR2020ZD17).\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "In the published article, there was an error in the Funding statement. We missed out an extra funder. The correct Funding statement appears below:The CERTAIN registry received funding from Astellas and Novartis. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication. All authors declare no other competing interests. The authors gratefully acknowledge the funding of the CERTAIN Registry by a grant from the Dietmar Hopp Stiftung, the European Society for Paediatric Nephrology (ESPN) and The German Society for Paediatric Nephrology (GPN). JKi acknowledges funding from a MRC NIHR fellowship (MR/V037900/1). CS acknowledges funding from the European Commision\u2019s Horizon 2020 Research and Innovation Programme (grant no. 952512). VK acknowledges funding from an NIHR Fellowship (PDF-2016-09-065) and as a Paul I. Terasaki Scholar.VK acknowledges funding from the National Institute for Health Research Blood and Transplant Research Unit in Organ Donation and Transplantation at the University of Cambridge in collaboration with Newcastle University and in partnership with NHS Blood and Transplant (NHSBT). The views expressed are those of the authors and not necessarily those of the National Health Service, the National Institute for Health Research, the Department of Health or National Health Service Blood and Transplant.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "There are errors in the Funding section. The correct Funding statement is: This research was supported by the Korean Association of Cardiopulmonary Resuscitation (no. 2021\u2013001) and the Soonchunhyang University Research Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. There was no additional external funding received for this study."} +{"text": "Funding statement. The grant number for funding from the Zhejiang Provincial Natural Science Foundation of China was incorrectly reported as \u201cNo. LR22C02003\u201d. The correct Funding statement appears below.In the published article, there was an error in the \u201cThis work was funded by the National Natural Science Foundation of China under Grant No. 32000234, and the Zhejiang Provincial Natural Science Foundation of China under Grant No. LR22C020003, the Major Science and Technology Special Project of Variety Breeding of Zhejiang Province (2021C02067-7), and the State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products under Grant No. 2021DG700024-KF202102\u201d.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Clarifying the integrated development mechanism of the National Fitness Program and National Health Program has positive and practical significance for the efficient implementation of the healthy China strategy. In order to analyze and study the integrated development mechanism of the National Fitness Program and National Health Program, we established a coupling-coordination measurement model. Based on two dimensions of space and time, we first analyzed the coupling degree between National Fitness System and National Health System in the Yangtze River Economic Belt region of China from 2016 to 2020, and studied and analyzed the coupling mechanism between the two. The research results show that the National Fitness Program and National Health Program have a development model of mutual promotion and mutual benefit. The National Fitness Program promotes the National Health Program by improving the fitness environment, increasing fitness participation, and health investment. The National Health Program optimizes environmental construction, health education, and healthy populations, thereby driving national fitness. Overall, there is stable development and balanced promotion in the integration of the National Fitness Program and National Health Program. In addition, the integrated development of the National Fitness Program and National Health Program is positively affected by economic level and regional marketization. The coupling- coordination model adopted in this study has practical value in measuring the coupling degree between the National Fitness System and National Health System. The results of this study also provide an important reference basis for the integrated and coordinated development of the National Fitness Program and National Health Program and the implementation of the Healthy China strategy. National fitness and national health have become important factors in social development and the improvement of people\u2019s well-being. The integrated development of national fitness and national health aligns with the global trend of health development. More and more countries and regions worldwide are treating national fitness and national health as important strategies, promoting people\u2019s health through measures such as providing fitness facilities, promoting health education, and strengthening health management. In response to this, General Secretary Xi Jinping of China has repeatedly emphasized the advocacy of a healthy and civilized lifestyle, the improvement of the health literacy of the entire population, and the deep integration of national fitness and national health during the period of 2016\u20132022. Meanwhile, the Chinese government has issued the \u201cNational Fitness Plan (2021\u20132025)\u201d and the \u201cSports Law of the People\u2019s Republic of China,\u201d which propose the implementation of a national fitness strategy, the establishment of a public service system for national fitness, and the encouragement and support of citizens\u2019 participation in fitness activities to promote the deep integration of national fitness and national health In terms of the connotation of the integration of the National Fitness Program and National Health Program, many scholars have started their research on the integration of the two systems by tracing the source of concepts. For examples, Lu believes(2) On the integration path of the National Fitness Program and National Health Program, much progress has been made. For examples, Lu formulat3) In terms of the mechanism of integrating the National Fitness Program and National Health Program, Lu In terms believesIn general, scholars have achieved fruitful results in the research on the integration relationship between the National Fitness Program and overall health, and have conducted well. They have analyzed and explored the integration relationship between the two from multiple perspectives of integration. However, there are two problems in current relevant research: first, the research has remained theoretical research on the National Fitness Program and National Health Program, and there is a lack of data exploration and verification of the integration relationship between the National Fitness Program and overall health. The coupling relationship between the two will affect the progress and effectiveness of comprehensive fitness and the achievement of the goal of national health. The second is that the scope of research is large, the theoretical coverage is broad, involving the entire national level, and there is a lack of specific comparative analysis of representative regions and different provinces and cities. Therefore, in the context of the healthy China strategy, it is particularly important to measure and evaluate the integration of the National Fitness Program and National Health Program through a combination of qualitative and quantitative research. Based on the analysis of the coupling effect between the National Fitness Program and National Health Program in various aspects, this paper constructs a model based on the two dimensions of space and time, analyzes the coupling and coordination relationship, and verifies the integration mechanism between the two. The research results of this paper have positive practical significance for achieving the efficient implementation of the healthy China strategy.2.2.1.Since the State Council promulgated the \u201cRegulations on National Fitness\u201d and the \u201cNational Fitness Plan 2011\u20132015),\u201d the National Fitness Program has been integrated into the overall planning of economic and social development of governments at all levels. After continuous efforts in recent years, the \u201cbig group\u201d work pattern of \u201cgovernment led, department coordinated, and the entire society participated\u201d has basically taken shape, and the effect has gradually become evident. Wei regarded,\u201d the Na2.2.The connotation of a Healthy China strategy is not only to ensure the health of the whole people, but also to ensure the overall health of the healthy environment, the healthy economy, and the healthy society. National health is also one of the main indicators to ensure the construction of a healthy city. The WHO defines a healthy city as an organic combination of healthy people, a healthy society, and a healthy environment. It has formulated 3 health indicators, 8 social indicators, 7 health service indicators, and 14 environmental indicators for a healthy city. In 2018, the China Health Commission released the \u201cChina Healthy City Evaluation Indicator System (2018 Version)\u201d , which c2.3.The concept of coupling originates from physics and refers to the existence of a relationship of mutual influence and interaction between two or more systems. In terms of the relationship between national fitness and national health, the two have similarities in service targets, convergence in core goals, and overlap in means and methods. Therefore, it is necessary to explore and analyze the integration relationship and mechanism between the two before building the model.2.3.1.The optimization of the national fitness environment can improve the development of the National Health System. The quality of the national fitness environment and the construction of the fitness environment are dialectically unified and complement each other. High environmental quality can greatly promote fitness activities. Conversely, low environmental quality is not conducive to the formation of residents\u2019 fitness habits, it is difficult to form a good fitness atmosphere and crowd participation rate, and environmental construction will also lag behind. Therefore, forming an active fitness atmosphere, rich sports life, strong fitness awareness, and a good fitness ecological environment can improve the quality of national fitness, promote the construction of a healthy environment, and promote the completion of the national health system. Starting from the \u201chard environment,\u201d increasing the availability and use of national fitness venues, facilities, and equipment, and popularizing the group fitness effect of community fitness can greatly improve fitness habits, promote the construction of a healthy environment, and further promote and improve the national health system.The quality and quantity of participation in national fitness can improve the over-all health system. Constantly improve the public service system for national fitness, ensure that everyone enjoys basic sports fitness services, and consolidate the basic conditions for national fitness to promote national health . The strThe volume ratio of national health investment can expand the National Health System. The health level of a country or region is usually measured by health input, which is reflected by human, material, and financial indicators of the health industry such as the total number of professional health personnel, the total number of medical and health institutions, and the total amount of health expenses .2.3.2.The construction of a healthy environment is the basic condition for promoting the development of national fitness. The development of national fitness needs to be supported by a good sports ecological environment. Through the construction and improvement of a healthy environment, such as increasing forest coverage and reducing smog days, more people can participate in physical exercise and expand the demand for sports market. National fitness has a high demand for healthy environmental conditions. Therefore, it is possible to start with managing the broken health environment, protecting and strengthening a good health environment, optimizing the construction of a healthy environment, and ultimately promoting the development of national fitness.The promotion of health education is a key factor in improving the awareness and needs of national fitness. With the increasing satisfaction of material pursuits, more and more people are paying attention to health education, not only physical health, but also psychological and social health. When the awareness of national fitness is constantly strengthened and demanded, it will be beneficial to expand the fitness community and promote the development of national fitness.per capita is bound to bring about an aging population, and the increase of the aging population will bring more health problems for the older adult. Developing healthy exercise for the older adult is particularly important. The youth group is the main group participating in physical exercise, mainly ensuring that the youth group exercises in a good healthy environment, and ensuring the stability and improvement of group development, in order to stabilize and ensure the national fitness system.The development of healthy people is an important guarantee for stabilizing the National Fitness System. The extension of life expectancy Based on the above analysis and discussion, the coupling mechanism model of the National Fitness Program and National Health program is shown in 3.3.1.3.1.1.Dimensionless processing can eliminate dimensional differences between variables, enhancing the stability and reliability of data analysis, improving the performance and effectiveness of models, reducing computational complexity, accelerating algorithm convergence, and enhancing the effectiveness of feature selection. The National Fitness System and National Health System are composed of several indicators. In order to unify the dimensions of each indicator, the raw data is standardized before data analysis. The calculation formula is as follows , 23:ijx means the original data of the j-th indicator of a province or municipality in year i, and ijy means the standardized data of the j-th indicator of a province or municipality in year i. min ijx is the minimum value of the i-th indicator and max ijx is the maximum value of the i-th indicator. The data of the dimensionless processing has the value of 0, which will interfere and affect the later calculation, so the data is panned. The calculation formula is as follows:The \u03b1 in 3.1.2.The advantages of the entropy method include: the absence of subjective weight setting, consideration of the correlation between indicators, strong flexibility and adaptability, easily interpretable and understandable results, applicability to multi-objective decision-making, as well as flexibility and adjustability. These features make the entropy method a commonly used multi-criteria evaluation approach that finds wide application in various fields and scenarios. Therefore, in this paper the entropy method is used to measure the index weights of national fitness and national health. The specific process is as follows \u201326:i-th year of the j-th indicator. The formula is as follows:Step 1: Homogeneous metrics are quantified. Measure the share of the ijp refers to the i-th year of indicator j.Where Step 2: The entropy value of the annual indicators of each region of the Yangtze River Economic Belt is calculated, and the proportion of each index is obtained. The entropy calculation formula for item j indicator is as follows:ih represents the specific entropy value of indicator j.Where j, the greater the ijy difference, the greater the impact on the results of specific provinces and cities, and vice versa. The specific calculation formula is as follows:Step 3: Calculate the difference coefficient for the indicator. For indicator jw. The calculation formula is as follows:Step 4: Determine the weight Step 5: Calculate the comprehensive measure value, that is, the calculation of the comprehensive score for the two systems of national fitness and national health. The calculation formula is as follows:su in Where 3.1.3.The coupling coordination model can organically combine factors from different domains or systems, forming a unified system framework. This systemic approach allows for a better capture of the complex relationships among interrelated factors and variables, thereby providing more accurate and comprehensive results and predictions. The coordinated advancement of the deep integration of national fitness and national health is a complex systemic project. Therefore, this study establishes a \u201ccoupling coordination\u201d calculation model for the National Fitness and National Health systems in order to analyze and evaluate the interconnectedness and coordination between these two systems. The formula for calculating the coupling coordination of the National Fitness System and National Health System is as follows \u201331:c represents the coupling degree between the National Fitness System and National Health System. f(x) and g(x) are the comprehensive evaluation index of the National Fitness System and National Health System of provinces and cities in the Yangtze River Economic Belt, respectively, obtained by the entropy method. And, \u03b1 and \u03b2 represent the pending coefficient, and R represents the coupling coordination. This paper believes that the importance of the two systems is the same, therefore, both \u03b1 and \u03b2 take 0.5, that is, \u03b1\u2009=\u2009\u03b2\u2009=\u20090.5. k is the adjustment coefficient, which is equal to 2. c\u2208, when c\u2009=\u20091, it indicates that national fitness and national health are in a highly coupled state; when c\u2009=\u20090, it means that there is no correlation between national fitness and internal elements of national health, disorderly development. Similarly, R\u2208. Coupling coordination levels are classified here The Yangtze River Economic Belt spans the east and west, with a population and economic total exceeding 40% of the country, and there is comparative analysis of data results; (2) As a new region for the implementation of China\u2019s new round of reform and opening up, it has global influence and is representative in data analysis. (3) Promoting the development of the Yangtze River Economic Belt is of great practical significance for the CPC Central Committee and the State Council to grasp and guide the New Normal of economic development, scientifically plan the new chess game of China\u2019s economy, and promote the coordinated development of regional economy, which is conducive to the industrial structure and urbanization layout along the Yangtze River, and promote economic efficiency and value added. The raw data on National Fitness and Nation Health of 11 provinces and cities in the Yangtze River Economic Belt was supplied in 4.4.1.per capita health cost is lower than the national average, the main health indicators of residents such as per capita life expectancy, infant mortality rate, and maternal mortality rate have been achieved, which are all better than the national average. Additionally, the data also supports the effectiveness of the implementation of the \u201cImplementation Plan for Health Lifestyle Action of the Whole Population (2017\u20132025)\u201d in Sichuan Province. By 2020, the health literacy level of residents in the province reached 20%, creating a collective action by the whole society and promoting a favorable atmosphere for the promotion and adoption of healthy lifestyle practices (ractices .4.2.The coupling coordination formula can be used to calculate the coupling value and coordination values of the National Fitness System and National Health System in 11 provinces and cities of China\u2019s Yangtze River Economic Belt from 2016 to 2020, as shown in The average coupling degree and coordination degree between the National Fitness System and National Health System in 11 provinces and cities of the Yangtze River Economic Belt in China are shown in 4.3.In order to further analyze the dynamic changes of the coupling degree and coordination degree between the National Fitness System and National Health System in various provinces and cities of the Yangtze River Economic Belt in China over time and space, we combined ArcGIS reclassification tool to visualize the data, and the results are shown in 4.4.per capita GDP ranking first in the country. It is a major economic province and also a major province in culture, sports, and health service industries. The development of the sports and fitness industry and the construction of healthy cities can basically form a virtuous cycle. This is closely related to Jiangsu Province continuously introducing new policies to promote the levels of the National Fitness System and National Health System. For example, the \u201cImplementation Plan for Implementing the Outline of Building a Sports Power\u201d issued by Jiangsu Province in 2020 explicitly states that by 2022, a new pattern of sports development will be formed, featuring strong government leadership, broad social participation, and a vibrant market. The physical literacy and health level of the population will continue to improve, new achievements will be made in sports reform and innovation, public services will be more balanced and sufficient, and the comprehensive strength of sports development will consistently rank among the top in the country, thus continuously increasing the contribution and influence of sports on economic and social development (The coupling coordination degree combines the coupling relationship between the National Fitness System and National Health System as a whole, which is more comprehensive and stable than the coupling results. The higher the coordination between the National Fitness System and National Health System, the higher the overall level of the two, and the greater the mutual promotion between the National Fitness System and National Health System. Conversely, one party hinders the development of the other party, forming a mutually contained and vicious cycle. From elopment . Overall5.This study combines qualitative and quantitative analysis to analyze the integration relationship and mechanism of fitness for all and health for all. Taking the panel data of 11 provinces and cities in the Yangtze River Economic Belt of China from 2016 to 2020 as an example, the comprehensive evaluation systems of the two systems are, respectively, constructed, and the evaluation and analysis are conducted according to the coupling coordination degree. The main research conclusions are as follows:Firstly, the National Fitness Program has improved the fitness environment, increased participation in fitness, and increased health investment, thereby promoting national health. The National Health Program has optimized environmental construction, health education, and healthy populations, thereby driving national fitness.Secondly, between 2016 and 2020, in the Yangtze River Economic Belt of China, Jiangsu and Zhejiang provinces ranked among the top three in the evaluation index of the National Fitness System and National Health System, belonging to the typical double high type. However, Chongqing and Guizhou have the smallest comprehensive evaluation index, indicating that they belong to the double low type of provinces and cities. This confirms that the integrated development of the National Fitness Program and National Health Program is positively influenced by economic level and regional marketization level.Thirdly, from 2016 to 2020, the average coupling degree between the National Fitness System and National Health System in the Yangtze River Economic Belt of China was between 0.8 and 1.0, which was in a high-level coupling stage. The average coupling coordination degree was between 0.4 and 0.7, which was in a medium to high degree of coordination coupling. This indicates that the coupling coordination pattern of the two systems has a certain degree of stability, and it also verifies that there is stable development and balanced promotion in the integration of the National Fitness Program and National Health Program.Fourthly, the high-level coupling stage between the National Fitness System and National Health System in the Yangtze River Economic Belt of China has increased from 9 provinces and cities to 11 provinces and cities between 2016 and 2020. The degree of coupling coordination has expanded from 4 provinces and cities to 9 provinces and cities, with Jiangsu Province reaching an extremely coupled coordination state. The higher the coordination between the National Fitness Program and National Health Program, the mutual promotion and mutually beneficial development of the two.The above results provide an important reference basis for the integrated and coordinated development of the National Fitness Program and National Health Program and the implementation of the Healthy China strategy.6.Contributions of this paper are as follows:Based on the composition and relational model of the National Fitness System and National Health System, a comprehensive evaluation system for the coupling degree of the National Fitness Program and National Health Program is constructed, which provides a beneficial supplement to the evaluation indicators of their coupling.The establishment of the coupling coordination calculation model for the National Fitness Program and National Health Program provides a practical calculation model for evaluating their coupling degree, offering important reference for research and practice in related fields.The statistical analysis of the coupling degree of the National Fitness Program and National Health Program in the Yangtze River Economic Belt of China reveals their integration mechanism, enriching the quantitative research on the integrated development of the National Fitness Program and National Health Program.In this paper, the measurement of the coupling coordination relationship between the National Fitness System and National Health System is carried out on the basis of multi-index evaluation system, which overcomes the shortcomings of the incomplete single index representation system, but there are also limitations that the construction of the index system is relatively flexible, which may lead to certain deviations in the final evaluation results. In addition, an in-depth analysis of the influencing factors of the spatial differences in the coupling coordination relationship between the National Fitness System and National Health System in 11 provinces and cities in the Yangtze River Economic Belt has not been conducted in the paper, and the spatial differences in the coupling and coordination relationship between the two systems may be affected by factors such as location and transportation, social economy, resource endowment and infrastructure. Therefore, the internal mechanism of the spatial difference features of the coupling coordination relationship between the 11 provinces and cities of the Yangtze River Economic Belt in China needs to be further studied. Moreover, due to the difficulty in collecting the latest data of national fitness in China, the evaluation index system for the coupling of the National Fitness System and National Health System constructed at present does not fully reflect the latest development trends, but only reflects a trend, whose latest data need to be further mined in the subsequent research.The original contributions presented in the study are included in the article/ CC contributions include writing original draft, investigation, and data curation. SY contributions include methodology, review, and editing. Both authors contributed to the article and approved the submitted version."} +{"text": "The authors regret that an incorrect version of product 74a in The authors also regret that incorrect details were given for ref. 91 in the original article. The correct version of ref. 91 is given below as ref. The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "In the published article, there was an error in the Funding statement. The funding statement for the Deutsche Forschungsgemeinschaft was displayed as \u201cTRR 369\u201d. The correct Funding statement appears below.\u201cThe present study was supported by the National Genome Research Network , the Pneumonia Research Network on Genetic Resistance and Susceptibility for the Evolution of Severe Sepsis , the Young Investigator Grant , the German Center for Lung Research and Stiftung AtemWeg . Work was supported by the Deutsche Forschungsgemeinschaft - TRR 359 - Project number 491676693.\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Correction: BMC Health Services Research 11, 217 (2011)https://doi.org/10.1186/1472-6963-11-217Following a report of a publications review jointly commissioned by the Health Research Authority and the University of Bristol, the authors would like to correct the following element of the ethics statement provided in the original article :The data used in this study were originally collected and collated in the National Outcomes Database for the purposes of evaluation of CFS/ME services. The collection of a subset of CFS/ME patient data as part of the national CFS/ME collaborative was confirmed to be service evaluation by the North Somerset & Bristol Research Ethics Committee under REC reference 07/Q2006/48, and in a letter dated 29 January 2007 the Chair of the Research Ethics Committee had previously confirmed (a) that it would not be necessary to apply for ethical permission to use the data being collected as part of service evaluation for the national CFS/ME collaborative and (b) that if in future these data were to be used as part of a research project, this would be agreeable.The original article has been"} +{"text": "The funders had no role in study design, the decision to publish, or preparation of the manuscript.There are errors in the Funding section. The correct Funding statement is: This work was supported by a Ph.D. studentship (2020.04778.BD) funded by the Portuguese Foundation for Science and Technology (FCT) and by the European Social Fund (ESF) attributed to Rita Pinto, and also by national funding from the Portuguese Foundation for Science and Technology (UIDB/00050/2020). FCT\u2019s website:"} +{"text": "Department of Pathogen Biology, Center for Tropical Disease Control and Research, School of Basic Medical Sciences and Life Sciences, Key Laboratory of Tropical Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, China\u201d. The new affiliation should be the first of all affiliations, and the affiliation numbers for authors Yiji Li and Tingting Li should be changed from 3 to 1. The final version of authors and affiliations are as shown above.In the published article, there was an error regarding the affiliation for author Zhuanzhuan Liu. As well as having affiliation 1, they should also have \u201cThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There are errors in the Funding section. The correct Funding statement is: Jun Song is supported by a Korea University Grant (K2127931), URL:"} +{"text": "Acknowledgements statement. Support received by CR from the Canada Research Chair program was omitted from the statement.In the published article, there was an error in the The corrected Acknowledgements statement appears below.CR was supported by the Canada Research Chair program (CRC-2018-00081).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Funding statement.In the published article, there was an error in the Original text: This work was supported by the National Natural Science Foundation of China under Grants 61971290.The Shenzhen City project support was missed by mistake. The correct Funding statement appears below.This work was supported by the National Natural Science Foundation of China under grant 61971290, and the Shenzhen Stability Support General Project (Category A) 20200826104014001.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Funding statement. The first grant number from the Fundamental Research Business Expenses of the Central Universities was incorrect. The incorrect grant number was 2021ZD001. The correct grant number is 2022YB013. The correct Funding statement appears below.In the published article, there was an error in the \u201cFundingThis research was funded by a grant from the Open Project of State Key Laboratory of Basic and Applied Aerospace Medicine (SMFA20K04); Fundamental Research Business Expenses of the Central Universities ; National Key R&D Program of the Ministry of Science and Technology (32071168); The authors report no involvement in the research by the sponsor that could have influenced the outcome of this work. The systematic review registration number PROSPERO 2019: CRD42018110290\u201d.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "There are errors in the Funding statement and the publisher apologizes for the errors. The correct Funding statement is as follows: This work was funded by the Deanship of Scientific Research at Najran University, Kingdom of Saudi Arabia, under the Research Groups funding program with the grant code number NU/RG/SERC/12/9. The authors express their gratitude for this support."} +{"text": "Correction: Molecular Imaging and Biology10.1007/s11307-023-01833-6The original online version of this article was revised to include the following funding information:This work was supported in part by the NIH HD 099090 to A.F. and by the Eunice Kennedy Shriver National Institute of Child Health & Human Development of the National Institutes of Health under Award Number T32HD087166, and Michigan State University to M.A.O.B. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.The original article has been corrected."} +{"text": "In \"Assessment of the Efficacy, Safety, and Effectiveness of Weight Control and Obesity Management Mobile Health Interventions: Systematic Review\" :e16899) the authors noted one clarification that should be added:The Acknowledgments section reads as:All authors contributed equally. The research for this paper was fully funded by the Instituto de Salud Carlos III from the Spanish Ministry of Science, Innovation and Universities, grant number PI16/01764.And will be changed to:All authors contributed equally. The research for this paper was fully funded by the Instituto de Salud Carlos III from the Spanish Ministry of Science, Innovation and Universities, grant number PI16/01764 co-funded by FEDER.The correction will appear in the online version of the paper on the JMIR Publications website on April 10, 2023 together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "There are errors in the Funding section. The correct Funding statement is: SHIP is part of the Community Medicine Research network of the University of Greifswald, Germany, which is funded by the Federal Ministry of Education and Research , the Ministry of Cultural Affairs as well as the Social Ministry of the Federal State of Mecklenburg-West Pomerania, and the network \u2019Greifswald Approach to Individualized Medicine (GANI_MED)\u2019 funded by the Federal Ministry of Education and Research (grant 03IS2061A). Genome-wide data have been supported by the Federal Ministry of Education and Research (grant no. 03ZIK012) and a joint grant from Siemens Healthineers, Erlangen, Germany and the Federal State of Mecklenburg- West Pomerania. The University of Greifswald is a member of the Cach\u00e9 Campus program of the InterSystems GmbH. This work was further supported by the project Superthyreose, funded by the German \u201cInnovationsfonds des Gemeinsamen Bundesausschusses\u201d (grant no. VSF2_2019\u2013167) AKB received funding from the BMBF and gratefully acknowledges an add-on-fellowship from the Joachim Herz Stiftung. LK acknowledges funding from the European Union , as well as the State of Lower Saxony and the Volkswagenstiftung . We acknowledge support for the Article Processing Charge from the DFG and the Open Access Publication Fund of the University of Greifswald. The funders had no influence on study design, data analysis, study interpretation, decision to publish, and writing of the manuscript."} +{"text": "He studied Chemistry at the National and Kapodistrian University of Athens, from where he graduated in 1954. He worked for two years at the Biochemistry Laboratory of the Hellenic Pasteur Institute, under the supervision of Zoi Mela-Ioannidis.On December 8, 2022, Efstratios (Stratis) Avrameas, a novel immunology researcher passed away in Paris. Stratis was born in Athens, on January 1In 1960, with a scholarship from the French government, he was accepted in the Microbial Chemistry Laboratory under the direction of Professor Pierre Grabar, at the Institute Pasteur, in Paris. Four years later (1964), he was awarded the title of Doctor of Biological Sciences (Doctorat d\u2019\u00c9tat), by Pierre and Marie Curie University, in Paris. In 1963, he was appointed Research Assistant by the National Center for Scientific Research (CNRS) of France, and worked in the Department of Protein Chemistry of the \u201cInstitute for Scientific Research on Cancer\u201d in VilleJuif, at first under the direction of Pierre Grabar and then under the supervision of Andr\u00e9 Lwoff, a Nobel prize laureate. In this laboratory, he rose up the hierarchy and became Research Director. In 1972, he was invited by Jacques Monod, another Nobel laureate, to return to the Institute Pasteur, in Paris where he headed the Immunocytochemistry Unit until his retirement, in 1999. In 1976, he was named Professor at the Pasteur Institute in Paris. From 1999 to 2002, he served in the position of Scientific Director at the Biotechnological Company DIATOS, founded by the French Pasteur Institute. His efforts aimed at developing proteins and peptides capable of transporting substances into cells with biological actions. After retiring from the Director\u2019s position at the Pasteur Institute in Paris, he returned to his homeland and worked voluntarily as a consultant both in the Immunology Laboratory of the Hellenic Pasteur Institute and the Immunology Laboratory of the Pathophysiology at the Athens University Medical School.1 This method replaced the radio-immunoassay (RIA) and thus, eliminated the exposure to radioactivity of employees working in research and diagnostic laboratories. Stratis, using the ELISA method, examined sera from healthy volunteers and demonstrated that these sera contained multitude of antibodies which recognised antigens of the human body (self-antigens autoantigens) and non-self antigens, namely, the gut microbiota. These antibodies had multireactivity. He called them natural autoantibodies.3 Based on these findings, he coined the theory of natural autoimmunity, in other words, the ability of the immune system to \u201cself-awareness\u201d before becoming able to recognise and fight the foreign invaders.5 Under physiological conditions, the auto-polyreactive natural autoantibodies have their active site blocked by the high amount of available self-antigens. In disease states, where self-antigens are released, the natural autoantibody network is a guardian that rapidly sequesters the released, from disease tissues, self-antigens. Moreover, these autoantibodies have vital homeostatic biological activities such as cellular repairing and enzymatic catalysis. Finally, based on our own experiments and Stratis\u2019 ones, we formulated the hypothesis that autoimmune diseases are the result of the hyper-function of natural autoimmunity against a specific cell or organ.6Professor Avrameas was a pioneer researcher of Immunology. He has published over 300 papers. At first, his research led to the coupling of an antibody to the enzyme peroxidase with the chemical agent glutaraldehyde. This achievement then, paved the way for the development of the enzyme-linked immunosorbent assay (ELISA).- Prize of the Foundation of Dr K. and Mme Peyre of the French Academy of Sciences (1974),- Celine Prize (1979),- Knight of the French Order of Merit (1980),- Gold Medal of the Indian Federation of Clinical Biochemistry (1981),- State prize (Prix d\u2019\u00c9tat) of the French Academy of Sciences (1985),- Corresponding Member of the Academy of Athens (1985),- Gold Award of the French Federation for the Promotion of Progress (1990),- Officer of the French National Order of Merit (1990),- Honorary member of the Hellenic Immunological Society (1991),- Honorary member of the National Medical Academy of Mexico (1995),- Medal of Honour of the French National Centre for Scientific Research (1997),- Honorary Research Director of the French National Centre for Scientific Research (CNRS) (1997),- Honorary Professor of the Pasteur Institute of Paris (1998),- Honorary Doctorate Degree of Athens University Medical School (2003),- Gold Cross of the Order of Honour from the President of the Hellenic Republic (2004).Stratis has been awarded the following significant prizes for his novel work:We, who had the honour and pleasure to work with Stratis, will all remember his honest and powerful personality, his desire to promote scientific knowledge through research and his passion for helping younger colleagues to acquire scientific knowledge.May his memory be eternal.Haralampos M. MoutsopoulosProfessor Emeritus and Member, Academy of Athens"} +{"text": "In the published article, there was an error regarding the affiliations for author Hira Singh. As well as having affiliation 1, they should also have Division of Vegetable Science, ICAR-Indian Agricultural Research Institute, New Delhi, India.An omission to the funding section of the original article was made in error. The following funding statement has been added:Authors are thankful to the ICAR-Indian Agricultural Research Institute, New Delhi for providing financial support and conduct of the research program of the PhD student, HS. The research work was partially funded by the NAHEP-CAAST programme of Indian Council of Agricultural Research.The authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "In the original published version of this article, the funding statement was printed incorrectly. This has now been corrected to:10.13039/100016714University of Sharjah, United Arab Emirates.\u201d\u201cThe accelerometers used in the study were sponsored by the College of Graduate Studies and the VC for Research and Graduate Studies Office, The publisher apologizes for the errors. Both the HTML and PDF versions of the article have been updated to correct the errors."} +{"text": "Chinese Journal of Bone and Joint Injury [Chinese Journal of Bone and Joint Injury and that there were significant differences in the listed authorship of the two manuscripts. This article is therefore retracted.Following publication, the authors of \u201cClinical Effect of Arthroscopic Resection of Extra-Articular Knee Osteochondroma\u201d by Chen et al. contactet Injury , in ChinJournal of Clinical Medicine. This retraction was approved by the Editor in Chief of The authors agreed to this retraction."} +{"text": "The following information is missing from the Funding statement: Parts of publication costs were covered by the Deutsche Forschungsgemeinschaft project number 491193532 and the Chemnitz University of Technology.The full statement reads as follows:This study was funded by German Research Foundation \u2014SFB 1410 Hybrid Societies awarded to F.H. Parts of the salary for Javier Baladron and Torsten Fietzek were covered by the fund above. Parts of publication costs were covered by the Deutsche Forschungsgemeinschaft project number 491193532 and the Chemnitz University of Technology. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "This article has been corrected at the request of the second author to change his affiliation from Arrowhead Regional Medical Center to\u00a0St. George's University School of Medicine, St. George's, GRD.The authors deeply regret that this error was not identified and addressed prior to publication."} +{"text": "There are errors in the Funding section. The correct Funding statement is: This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) and Korea Medical Device Development Fund (KMDF_PR_20200901_0103). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In the published article, there was an error where the Acknowledgments statement was not included. The missing Acknowledgments statement appears below.The research described in this paper is part of the MARS Initiative at Pacific Northwest National Laboratory. It was conducted under the Laboratory Directed Research and Development Program at PNNL, a multiprogram national laboratory operated by Battelle for the U.S. Department of Energy.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "There are errors in the Funding statement. The correct Funding statement is as follows: This work was carried out with the aid of a grant from UNESCO and the International Development Research Centre, Ottawa, Canada. The views expressed herein do not necessarily represent those of UNESCO, IDRC or its Board of Governors."} +{"text": "There are errors in the Funding statement. The correct Funding statement is as follows: YHL was supported by grants from the National Research Foundation (NRF) of Korea funded by the Ministry of Science and ICT [grant number: NRF-2021R1G1A1095517] as well as from Korea University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There is an error in"} +{"text": "There are errors in the Funding section. The correct Funding statement is: This study was part of a project on improving antibiotic allergy registrations, which was funded by the Dutch Ministry of Health, Welfare and Sport (Reference number 327952).The publisher apologizes for the error."} +{"text": "There are errors in the Funding section. The correct Funding statement is: This publication of this study is funded by the Universitas Indonesia PUTI 2023 Award: NKB-432/UN2.RST/HKP.05.00/2023. This research was also supported by the National Institute for Research and Development, Indonesian Ministry of Health which has been integrated into Indonesian National Research and Innovation Agency (BRIN). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There are errors in the Funding section. The correct Funding statement is: This article is part of the PhD dissertation approved at Iran University of Medical Sciences . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Funding statement. This study is supported by Health and Medical Research Fund (HMRF), Food and Health Bureau, Hong Kong, China (No. 18190691).The correct Funding statement appears below.In the published article, there was an error in the \u201cThis study is supported by Health and Medical Research Fund (HMRF), Food and Health Bureau, Hong Kong, China (No. 17180941)\u201d.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "There are errors in the Funding statement. The publisher apologizes for the errors. The correct Funding statement is as follows: The study was supported by Medical Research Council UK, Department of International Development (DFID), Economic and Social Research Council (ESRC) and Wellcome Trust under the Joint Health Systems Research Initiative (JHSRI) (Grant number: MR/NO15908) awarded to JG. It was also supported by the South African National Research Foundation (NRF) through the South African Research Chair (SARChi) in Health Systems and Policy awarded to the Centre for Health Policy, University of the Witwatersrand, and held by FG. All three authors received salary supplementation as part of the research funding from JHSRI. The specific roles of these authors are articulated in the \u2018author contributions\u2019 section. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Professor Sergey Bachurin received the Ph.D. degree in 1980 from Moscow State University. From 2006 to 2018, he remained the Director of the Institute of Physiologically active Compounds RAS. In 2003, he was elected as a member of the Russian Academy of Sciences. His research activity is related to the discovery of novel agents for neurodegenerative diseases treatment, in particular for Alzheimer's disease. He is the author of more than 250 articles in peer-reviewing journals and about 40 patents. His h-index is 33. Prof. Bachurin was an invited lecturer at the University of California (San Francisco), and at Tufts University (Boston)."} +{"text": "Funding statement. The incorrect funding number was used for the Medical Health Science and Technology Project of Zhejiang Provincial Health Commission. The correct Funding statement appears below.In the published article, there was an error in the"} +{"text": "Acknowledgements statement. Support received by CR from the Canada Research Chair program was omitted from the statement.In the published article, there was an error in the The corrected Acknowledgements statement appears below.CR was supported by the Canada Research Chair program (CRC-2018-00081).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "De la Rosa, and John Wiley and Sons. The withdrawal has been agreed due to a dispute over authorship and the conceptualization of some of the data.The above article, published online on 4 April 2023 in Wiley Online Library (wileyonlinelibrary.com) as an Accepted Article ("} +{"text": "The funding from Guangxi key research and development program was omitted. The correct Funding statement appears below:In the published article, there was an error in the Funding statement. The authors declare that this study received funding from Guangxi key research and development program No. (GK) AB18221080, The Basic Ability Enhancement Program for Young and Middle-aged Teachers of Guangxi (2020KY03036), Youth Foundation of Guangxi Medical University(GXMUYSF201918), Foundation of Guangxi Health and Family Planning Commission (Z20190581), Foundation of the Second Affiliate Hospital of Guangxi Medical University (hbrc202104). The funders were not involved in the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "This communication reflects the views of the RADAR-AD consortium and neither IMI nor the European Union and EFPIA are liable for any use that may be made of the information contained herein. Research of Alzheimer center Amsterdam is part of the neurodegeneration research program of Amsterdam Neuroscience. Alzheimer Center Amsterdam is supported by Stichting Alzheimer Nederland and Stichting Steun Alzheimercentrum Amsterdam. IK declares support for this work through the National Institute of Health Research and the Medical Research Council (Dementias Platform UK grant). CA\u2019s postdoctoral fellowship is funded by the Susan and Charles Berghoff Foundation. SG declares support for this work through the Italian Ministry of Health (Ricerca Corrente). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There are errors in the Funding section. The correct Funding statement is: The RADAR-AD project has received funding from the Innovative Medicines Initiative 2 Joint Undertaking under grant agreement No 806999. This Joint Undertaking receives support from the European Union\u2019s Horizon 2020 research and innovation programme and EFPIA and Software AG. See"} +{"text": "In January, 2023, the Science and Security Board of the Bulletin of the Atomic Scientists moved the hands of the Doomsday Clock forward to 90\u2009s before midnight, reflecting the growing risk of nuclear war commits each of the 190 participating nations\u201dto pursue negotiations in good faith on effective measures relating to cessation of the nuclear arms race at an early date and to nuclear disarmament, and on a treaty on general and complete disarmament under strict and effective international control\u201d UN,\u00a0. ProgresAny use of nuclear weapons would be catastrophic for humanity. Even a \u201climited\u201d nuclear war involving only 250 of the 13 000 nuclear weapons in the world could kill 120 million people outright and cause global climate disruption leading to a nuclear famine, putting 2 billion people at risk Bivens,\u00a0. A largehttp://www.ippnw.org).The health community has had a crucial role in efforts to reduce the risk of nuclear war and must continue to do so in the future Global Health Research Unit, is a member of the Independent Advisory Group, Collaboration for the Establishment of an African Population Cohort Consortium, and he was co\u2010chair of the InterAcademy Partnership, Climate Change and Health Working Group 2019\u20132022 and co\u2010chair of the Academy of Medical Sciences/Royal Society working group on \u201cA healthy future\u2014tackling climate change mitigation and human health together\u201d 2020\u20132021 . I. H. reports honoraria for several speaking engagements, all donated to Back from the Brink, the International Physicians for the Prevention of Nuclear War, or Physicians for Social Responsibility; travel/meeting support for Nobel Peace Laureates' Summit, the World Federation of Public Health Associations World Congress, and the UN Human Rights Commission Youth Summit; and he is a member of the steering committee of Back from the Brink and the International Steering Group of the International Campaign to Abolish Nuclear Weapons, a Board member of the International Physicians for the Prevention of Nuclear War and Physicians for Social Responsibility, and a Trustee of the Phillips Exeter Academy . M. G. M. O. R. reports research grants from the Dutch Research Council, NOW (grant number COMPL.21COV.001) and from the Netherlands Organisation for Health Research ZonMw (grant number 09120012010063) and he is Chair of the Dutch guideline committee on cognitive impairments and dementia. T. R. reports a contract with the Institute for Energy and Environmental Research (USA) for papers addressing the health and environmental consequences of nuclear testing in multiple locations, including Australia, French Polynesia, central Pacific, and China; honorarium from The Choisun Ilbo media group in South Korea for a lecture on nuclear weapons in 2022 and for nuclear weapons presentations from Hyogo Medical Practitioners Association (Japan), Peace Boat (Japan) and the University of Sydney; he was an expert witness on radiation and health for Environmental Justice Australia acting for Mine\u2010Free Glenaladale regarding proposed Fingerboards Mineral Sands Mine to the Victorian Government Fingerboards Inquiry and Advisory Committee; he is a member of RV3 Rotavirus Vaccine Development Scientific Advisory Board, Murdoch Children's Research Institute/Royal Children's Hospital; he is a member of the Committee of International Campaign to Abolish Nuclear Weapons Australia; he is a member of the Internet Peace Prize Award Committee ; he was a member of the Victorian International Humanitarian Law Advisory Committee, Australian Red Cross; he is a Board member of the Initiative for Peacebuilding, Faculty of Arts, University of Melbourne; he is an At\u2010large Board member of the International Physicians for the Prevention of Nuclear War; he was co\u2010president of the International Physicians for the Prevention of Nuclear War 2012\u201323; and he is honorary principal fellow, Melbourne School of Population and Global Health, University of Melbourne. P. Y. reports grants from Atea Pharmaceuticals; honoraria for lectures, presentations, and educational events from bioM\u00e9rieux and Pfizer Pharmaceuticals; fees for participation on an advisory board from Pfizer Pharmaceuticals; and he is a member of the Antimicrobial Stewardship Study Group Executive Committee (2022\u20132024) and the Clinical Practice Guideline Panel on Vaccinations in Immunocompromised hosts for the European Society of Clinical Microbiology and Infectious Diseases. C. Z. reports consulting fees for his role as senior adviser on the international journals project from the International Physicians for the Prevention of Nuclear War. The remaining authors declare no conflict of interest."} +{"text": "Finally, Staines and Barnhoorn invite the readers to submit abstracts for the next European Public Health Conference in November 2023 in Dublin. The theme of this year\u2019s conference is Our Food, Our Health, Our Earth: A Sustainable Future for Humanity.In this edition of the Public Health News, EUPHA\u2019s executive director addresses the European Health Data Space and the promises it holds for (public) health research as well as the challenges that lie ahead during the development of this infrastructure. Geller"} +{"text": "Acknowledgments statement. Support received by CR from the Canada Research Chair program was omitted from the statement.In the published article, there was an error in the The corrected Acknowledgments statement appears below.RRID:SCR_019195) and Lipidomics Core (RRID:SCR_019176), which receive financial support from the Faculty of Medicine and Dentistry and Canada Foundation for Innovation (CFI), and the latter also from the Natural Sciences and Engineering Research Council of Canada (NSERC) awards to contributing investigators. CR is supported by the Canada Research Chair program (CRC-2018-00081).We would like to thank all research participants, the staff from the HNRU, volunteers, undergraduate and summer students that contributed to this project. Experiments are performed at the University of Alberta Faculty of Medicine and Dentistry Flow Cytometry Facility (The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Nature Communications 10.1038/s41467-023-36599-6, published online 23 February 2023Correction to: The original version of this Article contained an error in the caption to the profile photos, which incorrectly read \u2018Priscilla Kolibea Mante is Co-Chair of the Global Young Academy, a neuroscientist and epilepsy drug expert at the Kwame Nkrumah University of Science and Technology in Kumasi, Ghana.\u2019 The correct version states \u2018Top left: Priscilla Kolibea Mante is Co-Chair of the Global Young Academy, a neuroscientist and epilepsy drug expert at the Kwame Nkrumah University of Science and Technology in Kumasi, Ghana. Top right: Encieh Erfani has been a member of the Executive Committee of the Global Young Academy since 2021 and is an assistant professor of Cosmology at the Institute for Advanced Studies in Basic Sciences in Iran. Middle left: Lisa Herzog is an alumna of the Global Young Academy and Dean of the Faculty of Philosophy at the University of Groningen in the Netherlands. Middle right: Jennifer Thomson is President of the Organization for Women in Science for the Developing World and Emeritus Professor in the Department of Molecular and Cell Biology at the University of Cape Town in South Africa. Bottom: Kaela Singleton is president-elect of Black in Neuro as well as a National Institute of Neurological Disorders and Stroke and Burroughs Wellcome Postdoctoral Enrichment fellow, completing her training in the Faundez lab at Emory University in the United States.\u2019https://globalyoungacademy.net/sciso/]\u201d.\u2019The original version of this Article contained an error in the response to question 6 by the GYA, which incorrectly read \u2018One project we have recently completed in science communication is SCISO, which stands for \u201cScience with Society\u201c.\u2019 The correct version states \u2018One project we have recently completed in science communication is SCISO, which stands for \u201cScience with Society [This has been corrected in both the PDF and HTML versions of the Article."} +{"text": "The correct Funding statement appears below.In the published article, there was an error in the Funding statement. This research was also funded by the \u201cWestern Grains Research Foundation\u201d and should be added to the list of funders. The previous Funding statement was \u201cThe authors acknowledge the financial support of the Dean\u2019s Scholarship and from the College of Graduate and Postdoctoral Studies (CGPS), University of Saskatchewan., the Saskatchewan Ministry of Agriculture, Saskatchewan Wheat Development Commission, and the Canada First Research Excellence Fund"} +{"text": "In \"Assessment of the Efficacy, Safety, and Effectiveness of Weight Control and Obesity Management Mobile Health Interventions: Systematic Review\" :e12612) the authors noted one clarification that should be added.In the Acknowledgments section it says:All authors contributed equally. The research for this paper was fully funded by the Instituto de Salud Carlos III from the Spanish Ministry of Science, Innovation and Universities, grant number PI16/01764.It should say:All authors contributed equally. The research for this paper was fully funded by the Instituto de Salud Carlos III from the Spanish Ministry of Science, Innovation and Universities, grant number PI16/01764 co-funded by FEDER.The correction will appear in the online version of the paper on the JMIR Publications website on April 10, 2023 together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "There are errors in the Funding statement. The correct Funding statement is as follows: This study was part of ZS\u2019s PhD project. It was funded by the Medical Research Council\u2019s Doctoral Training Programme (Funder reference number: 1658511) in the United Kingdom. Her attendance at the WHO CST Consultation Meeting in Xiamen, China, was funded by Autism Speaks. RH, BT, and CH received support from the National Institute for Health and Care Research (NIHR) for the SPARK project (NIHR200842) using UK aid from the UK Government. CH receives further NIHR support through the NIHR Global Health Research Group on Homelessness and Mental Health in Africa using UK aid from the UK Government. CH also receives support from WT grants 222154/Z20/Z and 223615/Z/21/Z. The views expressed in this publication are those of the authors and not necessarily those of the NIHR or the Department of Health and Social Care. The funders had no role in study design, data collection and analysis, the decision to publish, or the preparation of the manuscript."} +{"text": "There is error in the Funding statement. The correct Funding statement is as follows: This work was financially supported by the Deanship of Scientific Research at the King Faisal University, Saudi Arabia .The affiliation for the third author is incorrect. Shekh Md. Shajid Hasan Tusher is not affiliated with #2 but with #1: Department of Geography and Environmental Studies, University of Chittagong, Chittagong, Bangladesh."} +{"text": "In the published article, there was an error in the Funding statement. The funding statement for this project was:\u201cThis project was supported by the Centers for Disease Control and Prevention of the U.S. Department of Health and Human Services (HHS) as part of a financial assistance award totaling $3.8 million with 100 percent funded by CDC/HHS. The contents are those of the author(s) and do not necessarily represent the official views of, nor an endorsement, by Texas DSHS, CDC/HHS, or the U.S. Government. This project was also provided by the Southwest Center for Occupational and Environmental Health (SWCOEH), a NIOSH Education and Research Center, and awardee of (Grant No. 5T42) H008421 from the National Institute for Occupational Safety and Health (NIOSH)/Centers for Disease Control and Prevention.\u201dThe correct Funding statement appears below."} +{"text": "Prof. Caraci graduated MD in 2001 and he got his PhD degree in Neuropharmacology in 2009 at University of Catania. In March 2021 he was appointed Associate Professor in Pharmacology at University of Catania and Visiting Professor at University of Bordeaux in 2016/2017 as winner of the Visiting Scholar Grant 2016/2017. Prof. Caraci has worked in the field of drug discovery in depression, Alzheimer\u2019s disease and Down Syndrome with the aim to identify new pharmacological targets. Prof. Caraci has been studying the physiology and pharmacology of TGF-\u00df1 and its role in the pathophysiology of cognitive dysfunction in AD and Down Syndrome."} +{"text": "In the published article, there was an error in the Funding statement. The fund name is incorrect and the fund number is missing. The original text is: Programs for Science and Technology Development of Henan province. The correct Funding statement appears below.Programs for Science and Technology Development of Henan province No. 222102210078.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "There are errors in the Funding section. The correct Funding statement is: K.M.W was supported by grant from Business for Startup growth and technological development funded by Korea Ministry of SMEs and Startups in 2020. W.J.P was supported by grant from the National Research Foundation of Korea funded by the Korean government (MSIT). MSIT is the abbreviation of Ministry of Science and ICT. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In the published article, there was an error in the Funding statement. A funding agency was not included in the Funding statement. The correct Funding statement appears below.This research was partially supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health and Welfare, Republic of Korea (grant number: HI19C1343).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "There was an error in the funding statement. The correct funding statement is: This work was funded by a grant awarded to SY Zhang under the Key Construction Program of the National \"985\" Project and \"211\" Project. LH Yuan is supported by the Guangdong Academy of Sciences Scientific Research Fund (No.qnjj20091) and Guangdong Natural Science Fund (10451026001004389). G Jones, SJ Rossiter and SY Zhang received funding from a Biotechnology and Biological Sciences Research Council (BBSRC) China Partnering Award. The funders had no role in study design, design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In the Funding section, the number of an additional grant from the Czech University of Life Sciences Prague is incorrectly omitted from the Funding statement. The Funding statement should read: \u201cThis project was supported by grants from the Italian Ministry of Environment (MATTM), the Italian Institute for Environmental Protection and Research (ISPRA), the Czech University of Life Sciences Prague (IGA FTZ CZU 20135107 and 511120/1312/3108) and the International Visegrad Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "Multiple funding organizations and grants were incorrectly omitted from the Funding Statement. The Funding Statement should read: \"This study was supported by KAKENHI from the Ministry of Education, Science and Culture of Japan , Grant-in-Aid for Scientific Research on Innovative Areas (No. 24120522), the Society for Research on Umami Taste, and the Global COE Program in \"In Silico Medicine\" of Osaka University. The authors declare no conflict of interest with any financial organization regarding the manuscript. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "An introductory overview to the special issue papers on diffraction structural biology in this issue of the journal. An introductory overview to the special issue papers on diffraction structural biology in this issue of the journal. Journal of Synchrotron Radiation presents articles submitted in the context of the 4th International Symposium on Diffraction Structural Biology (ISDSB2013) held at the Fukiage Hall in Nagoya on 26\u201329 May 2013. It was the fourth in the series of ISDSB symposia initiated in 2003 by the Japan Society for the Promotion of Science (JSPS) and specifically by the University\u2013Industry Cooperative Research Committee (#169) chaired by Professor Noriyoshi Sakabe. The previous conference was held in France, close to Paris at Paris-Sud University (Orsay) and the synchrotron radiation facility SOLEIL, chaired by the late Roger Fourme. ISDSB2013 came back to Japan again.This issue of the The basic concept of the ISDSB symposia is, firstly, to bring together researchers using diffraction and crystallography, and more generally interactions of X-rays, electrons and neutrons with matter, in the study of structural biology, and, secondly, within this domain, to facilitate the interaction between academic and industrial researchers. The interfaces with other active fields in biological ultra-structure using microscopies and spectroscopies (in particular nuclear magnetic resonance) are also respected as vital for the growth of a more systematic understanding of biological function based on structure. The scientific topics covered in ISDSB2013 included the following sessions: (1) synchrotron radiation and free-electron lasers; (2) new methodology and instrumentation; (3) and (4) drug design; (5) electron microscopy; (6) tomography and imaging; (7) neutron diffraction and hydration structure; (8) membrane proteins and macromolecular complexes; (9) protein structure and dynamics. A total of 207 participants, with a number of young scientists and PhD students, from 14 countries took part. The speakers included two Nobel Prize winners, Thomas Steitz and Brian Kobilka, three plenary lecturers and 31 invited speakers. A poster session was also held and 80 posters were presented. Thirteen commercial and industrial companies presented exhibitions, and a sponsored lunch seminar was also given.The symposium was supported jointly by the #169 Committee of JSPS and by many industrial companies as well as generous individuals. We wish to express our sincere thanks to the University\u2013Industry Cooperation and Research Program Division of the JSPS, to the Industry Club of Japan for their financial contribution through JSPS to print this issue, to the Naito Foundation for supporting the travel fees, and to all companies and persons who contributed their donations and cooperation."} +{"text": "The correct funding information is: \u201cThis work was sponsored by an unrestricted grant of the Osteoporosis Platform of the Swiss Society for Rheumatology. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There was an error in the Funding statement.The correct Funding statement is: The work was supported by the National Natural Science Foundation of China (30971264). The funders had no role in study design, data collection, analysis, decision to publish or preparation of the manuscript."} +{"text": "This project is a community-level study of equity of access to eye health services for Indigenous Australians.The project used data on eye health services from multiple sources including Medicare Australia, inpatient and outpatient data and the National Indigenous Eye Health Survey.The analysis focused on the extent to which access to eye health services varied at an area level according to the proportion of the population that was Indigenous . The analysis of health service utilisation also took into account age, remoteness and the Socioeconomic Indices for Areas (SEIFA).The rate of eye exams provided in areas with very high Indigenous populations was two-thirds of the rate of eye exams for areas with very low indigenous populations. The cataract surgery rates in areas with high medium to very high Indigenous populations were less than half that reference areas. In over a third of communities with very high Indigenous populations the cataract surgery rate fell below the World Health Organization (WHO) guidelines compared to a cataract surgery rate of 3% in areas with very low Indigenous populations.There remain serious disparities in access to eye health service in areas with high Indigenous populations. Addressing disparities requires a co-ordinated approach to improving Indigenous people\u2019s access to eye health services. More extensive take-up of existing Medicare provisions is an important step in this process. Along with improving access to health services, community education concerning the importance of eye health and the effectiveness of treatment might reduce reluctance to seek help. Equitable and accessible health services form a key component of equitable health outcomes. Addressing inequities in health services is particularly important in eye health where the disadvantage of Indigenous Australians is unequivocal. The rate of low vision in Indigenous adults is 2.8 times rates in the general population and the rate of blindness 6.2 times higher than general population rates.A core concept in equitable health care is distribution according to level of need for services, without regard to characteristics that do not inform these needs-4. InequOur project examines equity of access to eye health services at a community level by examining the relationship between the percentage of Indigenous people living in an area, socioeconomic status and remoteness, access to ophthalmic and optometric services and the professionals that provide them.The data request was reviewed by Medicare Australia to ensure that the proposed use of the data was ethical and compliant with relevant legislation including the Information Privacy Principles under section 14 of Privacy Act 1988, Health Insurance Act 1973, National Health Act 1953, Health identifiers Act 2010 and Freedom of information Act. The provision of hospital data from State and Territories was similarly reviewed by the relevant data custodians including Corporate Data Management & Reporting; Information Management Section, Australian Capital Territory (request 2762); Health Demand and Performance Evaluation, New South Wales Health; Acute Care Information Unit, Northern Territory Department of Health and Families; Statistical Output, Health Statistics Centre, Queensland Health; Operations Division, South Australia Health; Health Information Provision, Victorian Department of Human Services (request 2601); Data Collection and Analysis - Statutory and Non-Admitted; Information Management and Reporting ( Request 3a_2009OP), West Australian Department of Health Service Review and Enhancement; Department of Health Services, Tasmania.Privacy Act 1988, Information Privacy Act 2000 and the Health Records Act 2001 in Victoria, Health Records and Information Privacy Act 2002 No 71 and Privacy and Personal Information Protection Act 1998 in New South Wales, Health Records (Privacy and Access) Act 1997 in Australian Capital Territory, the Information Privacy Act 2009 in Queensland, Personal Information Protection Act 1991 in Tasmania, Information Act 2002 in Northern Territory, Information Privacy Bill 2007 in Western Australia and Information Privacy Principles 2009 in South Australia.Relevant legislation for standards for information collection, storage, access, transmission, disclosure, use and disposal included the Commonwealth Data on the geographic distribution of ophthalmology practices was obtained from the Royal College of Ophthalmologists (n\u2009=\u20091058) membership in 2008. Data on the geographic distribution of optometry practices was obtained from the 2008 electronic white pages (n\u2009=\u20096270). The number of practitioners is estimated to be equal to the number of offices. This will overestimate practitioners if offices are only visited and operated periodically.Medicare data were obtained for services provided by optometrists and ophthalmologists . Data covered the period from 2004/05-2007/08 . The utilisation data was broken down by age and statistical subdivision (SSD). The SSD is a general purpose geographical unit determined by the Australian Bureau of Statistics. In aggregate, SSDs cover Australia without gaps or overlaps. For the 2001 Census there were 207 SSDs defined throughout Australia. Medicare is available to people who reside in Australia, excluding Norfolk Island, if they hold Australian or New Zealand citizenship or have been issued with or applied for a permanent visa. Medicare provides access to free treatment as a public patient in a public hospital, free or subsidised treatment by practitioners and free treatment by providers who bulk bill.Data on attendances at outpatient from eye clinics were also collected from each State and Territory by SSD. Data on paediatric attendances were not available from one region of Western Australia because of privacy concerns. Attendance at this clinic was estimated based on the ratio between adult and child hospital separations attendances in the rest of WA. The only complete year of data available for all states and territories was for 2007/08. Outpatient data do not include data on either the age or the Indigenous status of patients seen.Each State and Territory was contacted to identify any other major programs that would not be captured using Medicare and hospitalisation data. While some small additional programs were identified in Queensland, there was no evidence of other major initiatives.Hospital inpatient data for cataract related Australian National Diagnosis Related Groups for public and private hospitals were obtained from New South Wales, Victoria, Queensland, South Australia and Tasmania by SSD and Aboriginality.The availability of inpatient data from Western Australia and the Northern Territory was limited because of concerns of privacy. Western Australian data was provided by procedure and region. Rates of use in each SSD were then estimated based on population size. Northern Territory data for eye procedures was provided at Territory level. The distribution of services across the Territory was then estimated using the overall distribution of hospital procedures.Inpatient data does include an Indigenous identifier but this suffers from well-documented problems of under-enumeration. The extPopulation projections by age and part of the state for the whole population were obtained from SuperTABLE 4.3.1 Build 10. PopulatIntercooled Stata v10 was used to conduct a panel poisson regression using SSDs as the unit of analysis. The dependent variables for the primary care analysis were eye exams provided by optometrists and ophthalmologists through Medicare and hospitals. The dependent variables in the hospital analysis were the supply of services for cataracts through private and public hospitals. The independent variables were year and the percentage of Indigenous people living in each area. The analyses were run adjusting for age, remoteness and area socioeconomic advantage, disadvantage and socioeconomic educational status. Remoteness was coded into categories . Socioeconomic data were based on the Socio-economic Indexes for Areas (SEIFA). The SEIRates of cataract surgery were also compared to the national average and World Health Organization (WHO) guidelines. Most deTableFiguresTableTableA number of areas had cataract surgery below the levels generally recommended by the WHO to reduce cataract blindness in Africa. Around 40 percent of areas with very high Indigenous populations had cataract surgery rates below WHO recommended guidelines. This compares to around 6 percent in reference areas Table. Around The fragmented funding and service provision in the Australian health system creates a major challenge for health services research and the assessment of equity at a system level. We have drawn together information across sectors to provide a comprehensive assessment of eye health services for Indigenous Australians. The study supports the notion that socioeconomic status, ethnicity and geography can all contribute to inequalities in eye health-25. It hAs an ecological study, this research represents an indirect examination of Indigenous eye care access and utilisation. While there are other factors that affect this distribution, including measures of remoteness and socioeconomic disadvantage, there is little doubt that areas with a larger proportion of Indigenous Australians are the most disadvantaged in terms of access to eye health services at a primary care level. The rate of total eye exams provided in areas where the Indigenous population was very high was two-thirds of the rate of eye exams for areas where the Indigenous population was very low. Areas with very high Indigenous populations constituted about two-thirds of areas where the provision of eye health services was significantly below the national average.Broadening the range of health professionals able to obtain reimbursement through Medicare is a key strategy of the reform of the Australian health system. OptometIt is concerning that many areas had rates of cataract surgery that fell below the WHO guidelines. It is eThere is currently heated debate around reduction in the Medicare reimbursement rates for cataract and its implications for service provision. IncreasThe results suggest that despite a number of government initiatives to improve Indigenous people\u2019s utilisation of eye health services there remain significant inequities in access. Even though Australia is a developed country, there was evidence that treatment for cataract in some areas with large Indigenous populations fell below WHO guidelines developed for Africa. Developing a targeted co-ordinated approach to address these issues is a challenge in an environment of complex service provision. More extensive take-up of existing Medicare provisions would be an important step in this process. The National Indigenous Eye Health Survey data suggest that along with improving access to health services, community education around the importance of eye health and the effectiveness of treatment might reduce reluctance to seek help.ATSIC: Aboriginal and Torres Strait Islander Commission; AIHW: Australian Institute of Health and Welfare; SEIFA: Socioeconomic Indices for Areas; SLA: Statistical Local Area; SSD: Statistical subdivision; WHO: World Health Organization.The authors declare that they have no competing interests.MK contributed to the design of the project, drafted the paper and conducted the final analysis. AF contributed to the design of the project, collected the data, conducted preliminary analysis and commented on drafts. HT contributed to the design of the project and commented on drafts. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2415/12/51/prepub"} +{"text": "There was information missing from the funding section. The complete, correct funding statement is: This study was funded by Pain Therapeutics who, at the time of the study, had the license for RIT of HIV. The funders paid for research personnel effort, supplies and laboratory animals through industrial research agreements with Albert Einstein College of Medicine and with University of California Los Angeles. Currently Pain Therapeutics do not have the licensing rights for this technology. This work was also supported in part by the Bill and Melinda Gates Foundation and Center for AIDS Research at the Albert Einstein College of Medicine and Montefiore Medical Center funded by the National Institutes of Health (NIH AI-51519). Pain Therapeutics participated in the study design, analysis, decision to publish, and preparation of the manuscript. Other funders did not participate in the study design, analysis, decision to publish, and preparation of the manuscript."} +{"text": "The Funding information was incorrect. The correct Funding is: This material is based upon work supported in part by the National Science Foundation under Grant #0962805. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There was an error in the Funding statement. The correct version of the statement is:This work was supported by grants from 973 program of China (2010CB529100)\uff0cand the Natural Science Foundation of China , as well as by Peking University People\u2019s Hospital Research and Development Funds (RDB2012-04). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The Funding line should read: \"The authors acknowledge funding for this research from WWF-Australia. RLP acknowledges support from the Australian Research Council. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There were errors in the Funding section. The correct funding information is as follows:This work was supported by the Bill & Melinda Gates Foundation through Grand Challenge 6; and the EDCTP through the African European TuberculosisConsortium and the Trials of Excellence in SouthernAfrica . Dr Chegou received financial support from theClaude Leon Foundation and the South African MRC during the writing up of this work. Thefunders had no role in study design, data collection and analysis, decision to publish, orpreparation of the manuscript."} +{"text": "The Editor of this article was mistakenly omitted from the final document. The publisher apologizes for the error. The Editor is:Jinfa Zhang, New Mexico State University, USA."} +{"text": "There was an error in the Funding section. The correct funding information is as follows: This study was supported by National March of Dimes and Heart Foundation grants to MEW and KMM. LAG was supported by a National Heart Foundation of Australia Biomedical Scholarship. KMM was supported by an National Health and Medical Research Centre Fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "This presentation will demonstrate how public mental health systems can deliver accessible, effective and client-centred eating disorder treatment. It will provide an overview of service development over time aimed at increasing workforce capacity, responsiveness of service provision, improved access and better integration of all essential elements of care. The presentations will describe and discuss this service delivery initiative in the Southern region of Melbourne, looking at barriers, enablers and implications for policy and planning. An overview of the development of an integrated mental health and paediatric service for young people with eating disorders at Alfred Health Child and Youth Mental Health Service will be provided. Data on service provision and outcomes will be discussed, along with a case example illustrating a typical case presenting for treatment at this service.Children and Youth Treatment and Service Development stream of the 2013 ANZAED Conference.This abstract was presented in the"} +{"text": "To understand the full burden of a health condition, we need the information on the disease and the information on how that disease impacts the functioning of an individual. The ongoing revision of the International Classification of Diseases (ICD) provides an opportunity to integrate functioning information through the International Classification of Functioning, Disability and Health (ICF).Part of the ICD revision process includes adding information from the ICF by way of \u201cfunctioning properties\u201d to capture the impact of the disease on functioning. The ICD content model was developed to provide the structure of information required for each ICD-11 disease entity and one component of this content model is functioning properties. The activities and participation domains from ICF are to be included as the value set for functioning properties in the ICD revision process.The joint use of ICD and ICF could create an integrated health information system that would benefit the implementation of a standard language-based electronic health record to better capture and understand disease and functioning in healthcare. Describing and understanding the relationship between disease and functioning requires the use of two of the World Health Organization\u2019s classifications systems: the International Classification of Diseases (ICD) and the th revision (ICD-11) [The ICD is undergoing its 11(ICD-11) wherein The process of revising the ICD is coordinated through Topic Advisory Groups (TAGs), each of which is responsible for different content areas. Responsible for the appropriate integration of the functioning properties is the Functioning Topic Advisory Group (fTAG), which consults with each of the TAGs regarding how to deal with functioning properties for their assigned ICD entities.Activity is defined in the ICF as the \u201cexecution of a task or action by an individual\u201d, while participation is defined as \u201cinvolvement in a life situation\u201d [The ICD-11 Content Model by content experts worldwide in three steps: [Before ICD-11 is completed, functioning properties will need to be populated for each ICD code. This task of population is being done and coordinated using the web-based e steps: if an ade steps: use evide steps: .Obtaining information about disease entities and their impact on functioning is not entirely new in the field of medicine and health. While the consideration of the disease and its impact on functioning has been in place, or at least acknowledged, for a long time, there reThe ICD-11 is due to be launched in 2015, and steps toward that goal are being pursued. Certainly there are challenges on our way, but there are also opportunities that are presented for users in the clinical and research communities to actively contribute in this huge endeavor by WHO and its collaborators worldwide. The unified ICD-ICF in the ICD-11 will allow for consistent terminologies to be used and to be harmonized across ICD and ICF and will provide holistic information about a disease entity and its impact on the functioning of an individual. Efforts are also currently being taken to facilitate the identification of the overlaps for ICD-11 disease entities and their titles with their conceptual equivalent in the ICF towards harmonization of ICD and ICF.The joint use of the ICD and ICF towards an integrated health information model would, in our opinion, benefit medicine and health systems and would support the push for the implementation of a standard language-based electronic health record system towards better health services planning and reimbursement.fTAG: Functioning Topic Advisory Group; iCAT: International Collaborative Authoring Tool; ICD: International Classification of Diseases; ICF: International Classification of Functioning, Disability and Health; TAG: Topic Advisory Group; WHO: World Health Organization.The authors declare that they have no competing interests.All authors provided concept/idea, consultation, and writing, and reviewed the manuscript before submission. All authors read and approved the final manuscript.RE is Assistant Professor, Department of Physical Therapy, School of Allied Health Professions Louisiana State University Health Sciences Center, New Orleans LA USA; adjunct research scientist at the ICF Research Branch in cooperation with the WHO Collaborating Centre for the Family of International Classifications in Germany (DIMDI), Nottwil, (Switzerland); and the Swiss Paraplegic Research (SPF), Nottwil, Switzerland.NK is technical officer at World Health Organization, Classifications, Terminologies and Standards (CTS), Department of Health Statistics and Informatics (HSI), Geneva, Switzerland.CK is with the US Department of Health and Human Services, Office of Health Policy Washington D.C., USA.MMRN is technical officer at World Health Organization, Classifications, Terminologies and Standards (CTS), Department of Health Statistics and Informatics (HSI), Geneva, Switzerland.GS is director of the ICF Research Branch in cooperation with the WHO Collaborating Centre for the Family of International Classifications in Germany (DIMDI), Nottwil, (Switzerland) and the Swiss Paraplegic Research (SPF), Nottwil, Switzerland; is Professor and Chair at the Department of Health Sciences and Health Policy, University of Lucerne, Lucerne, Switzerland.TBU is head of WHO\u2019s Family of International Classifications, Geneva, Switzerland.R Escorpizo is an employee of the Louisiana State University Health Sciences Center (LSUHSC). This article was developed in his professional capacity and does not necessarily represent the views of LSUHSC.C Kennedy is an employee of the Office of the Assistant Secretary for Planning and Evaluation (ASPE), Department of Health and Human Services (HHS). This article was developed in her professional capacity and does not necessarily represent the views of ASPE or HHS.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/13/742/prepub"} +{"text": "Grants to the second and third authors (AV and JP) are incorrectly omitted from the Funding statement. The Funding statement should read: \u201cThe PC lab was supported by the Swiss National Foundation for Scientific Research (grant 31003A-135789), the Doerenkamp-Zbinden Foundation and the Fondation Egon Naef pour la Recherche in Vitro. The JP lab was supported by grants from the Swiss National Science Foundation. WCL was partially funded during this project with a Telethon Action Suisse grant and AV was funded by an EMBO Long Term Fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "There was an error in the funding statement. The correct funding statement is: \"This work was supported by the Hungarian National Research Fund (PD83444), by the J\u00e1nos Bolyai Research Scholarship of the Hungarian Academy of Sciences (for MI and CSA), the Czech Ministry of Education, Youth and Sports (grant awards LC06004 and OC08025) and the European Regional Development Fund . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "There is information missing from the Funding statement. The complete funding statement is: This study was financially supported by the SNAC project , the GENOBACT project and was also supported by the Doctoral School on the Agro-Food System (Agrisystem) of the Universit\u00e0 Cattolica del Sacro Cuore . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There was an error in the Funding statement. The correct version of the Funding statement is available below.This work is funded by the grants from the Ministry of Science and Technology of China (S & T major Program: No. 2012ZX1004701-001-002), the Basic Research Program of China , the National Nature Science Foundation of China , and Beijing Natural Science Foundation (No. 5112022). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\" Two grants from the National Science Council were incorrectly omitted. The Funding Statement should read: \"This project was supported by VGH92-378 from the Taipei Veterans General Hospital ("} +{"text": "With limited time to achieve the Millennium Development Goals, progress towards improving women's and children's health needs to be accelerated. With Africa accounting for over half of the world's maternal and child deaths, the African Union (AU) has a critical role in prioritizing related policies and catalysing required investments and action. In this paper, the authors assess the evolution of African Union policies related to women's and children's health, and analyze how these policies are prioritized and framed.The main method used in this policy analysis was a document review of all African Union policies developed from 1963 to 2010, focusing specifically on policies that explicitly mention health. The findings from this document review were discussed with key actors to identify policy implications.With over 220 policies in total, peace and security is the most common AU policy topic. Social affairs and other development issues became more prominent in the 1990s. The number of policies that mentioned health rose steadily over the years .This change was catalysed by factors such as: a favourable shift in AU priorities and systems towards development issues, spurred by the transition from the Organization of African Unity to the African Union; the mandate of the African Commission on Human and People's Rights; health-related advocacy initiatives, such as the Campaign for the Accelerated Reduction of Maternal Mortality in Africa (CARMMA); action and accountability requirements arising from international human rights treaties, the Millennium Development Goals (MDGs), and new health-funding mechanisms, such as the Global Fund to Fight AIDS, Tuberculosis and Malaria.Prioritization of women's and children's health issues in AU policies has been framed primarily by human rights, advocacy and accountability considerations, more by economic and health frames looking at investments and impact. AU policies related to reproductive, maternal, newborn and child health also use fewer policy frames than do AU policies related to HIV/AIDS, tuberculosis and malaria.We suggest that more effective prioritization of women's and children's health in African Union policies would be supported by widening the range of policy frames used and strengthening the evidence base of all policy frames used. In addition, we suggest it would be beneficial if the partner groups advocating for women's and children's health were multi-stakeholder, and included, for instance, health care professionals, regional institutions, parliamentarians, the media, academia, NGOs, development partners and the public and private sectors. With limited time remaining to achieve the Millennium Development Goals (MDGs), there needs to be accelerated progress to achieve MDGs 4 and 5 and improve women's and children's health . This isAfrican countries have a leadership role in developing and implementing the required policies and programmes to achieve progress in women's and children's health. However, many countries are faced with multiple development priorities and limited resources. Improving the health of women and children will require national commitment to the health agenda at the highest level.Regional bodies such as the African Union (AU) set regional policy priorities. They also increasingly influence national and global policies as the need for linking global and national initiatives and for partner coordination increases .The African Union has already shaped national health priorities. For instance, in 1999, the African Union became the first and only regional body to pass a charter on the rights of the child. In 2006, the Protocol to the African Charter of Human and People's Rights on the Rights of Women in Africa became the first convention to mandate state provision of comprehensive reproductive and sexual health services . This reAfrican Union policies have also been instrumental in persuading some African policy-makers to recognize HIV/AIDS , leadingAs with HIV/AIDs and other issues, the African Union can be an important stakeholder in promoting commitments to improve the health of women and children and ensuring accountability for their implementation. The Campaign for the Accelerated Reduction of Maternal Mortality in Africa, led by the African Union, has been launched in 34 countries . In 2010This paper analyses how the African Union has prioritized policy-making on women's and children's health, and outlines the implications of the trends and how policy-making can be strengthened.The main method used in this policy analysis was a document review of all African Union policies developed from 1963 to 2010, focusing specifically on policies that explicitly mentioned health. The findings from this document review were discussed with key experts to identify policy implications.To conduct the document review, all African Union Head of State Assembly Outcome Documents were obtained from the African Union online archive . \"Outcomper se: one for African Union governance and one for cross-cutting policies. The combined list of the eight African Union portfolios and the two additional categories are :The 884 policies found in the 55 outcome documents were categorized according to the 8 portfolios of the African Union Commission Table . Policie1. African Union Governance: Core Business Management, Development of Strategies, Implementation Facilitation, Costs, Appointments, New Structures.2. Cross-cutting: Development, Poverty Reduction, Regional Cooperation, and any declarations that cover issues in more than one category.3. Economic affairs: Economic Integration, Monetary Affairs, Private Sector Development, Investment and Resource Mobilisation.4. Human resources, science and technology: Education, Information and Communication Technology, Youth, Science and Technology, Human Resources.5. Infrastructure and energy: Energy, Transport, Communications, Infrastructure and Tourism.6. Peace and security: Conflict Prevention, Management and Resolution, and Terrorism Issues.7. Political affairs: Democracy, Human Rights, Good Governance, Electoral Institutions, Humanitarian Affairs, Civil society Organisations, Refugee Matters.8. Rural economy and agriculture: Rural Economy, Agriculture and Food Security, Livestock, Environment, Water, Natural Resources and Desertification9. Social affairs: Health, Children, Drug Control, Gender, Labour and Employment, Sports and Culture, Migration10. Trade and industry: Trade, Industry, Customs and Immigration MattersThe numbers of policies developed in each of these categories were compared across time to determine historical trends in AU policy-making.per se, resulting in a total of 55 health policies that were included in this analysis.In order to identify health-related policies, a full text keyword search of the 55 outcome documents using the term \"health\" was conducted. This search yielded 56 policies from across all portfolios. One of these policies was excluded as the reference was to 'healthy management' practices and not to health Some specific health policies-for example on Polio or HIV/AIDS-were not captured in the keyword search using \"health\". Traditionally all health-related policies are located within the Social Affairs portfolio. To ensure that all policies related to health were captured, the authors further reviewed and categorized the 102 policies within the Social Affairs portfolio based on their reference to specific health topics e.g. AIDS/TB/Malaria, reproductive, maternal, newborn and child health (RMNCH), and to women's and children's rights to health and development .In order to identify how and why women's and children's health is prioritized in African Union policies, the authors analysed all health-related policies, according to the main arguments or \"frames\" used. Based on a review of previous policy analyses -22 a preAccountability: legal, policy, and monitoring requirements, including human rights, and African Union and Millennium Development Goals reporting.1. Advocacy: prioritization statements from influential stakeholders, media coverage, events, and advocacy campaigns.2. Economic: social and economic development, including trade, productivity, cost-effectiveness analysis, efficiency and trade-offs.3. Health: scientific evidence and technical information on health outcomes, effective interventions and health systems.4. Other: anything falling outside of frames 1-4, such as cultural norms and references.5. In additional to these frames, and in line with Walt's framework of policy analysis, which promotes a review of content and context of policies, as well as a review of actors and processes that influence policy-making , the codUsing these coding parameters, two analysts first coded a random sample of 5 of the 81 health-related policies. The coding was done in Excel. The inter-rater reliability was assessed, and through an iterative process, the analysts identified discrepancies in coding, refined the analytical frames in discussion with the research team and retested inter-rater reliability. The final intercoder agreement was 0.80, which is considered acceptable .The authors also compared the policy frames used in AU policies related to MDGs 4 and 5 on reproductive, maternal and child health, with AU policies on MDG 6 related to HIV/AIDS, tuberculosis and malaria.To validate the findings and discuss policy implications, results of the document review were discussed with a purposive sample of 10 key actors involved in the African Union and in health policy and processes on the continent.The actors included high-level representatives of the African Union, ministries of health and non-governmental organizations working on African Union issues.Interviews were conducted in a semi-structured format with written notes (Annex 2-list of questions). Discussions included a report by the interviewer of the preliminary findings of the document review and information from key actors on the reasons underlying review findings, policy implications of these findings and recommendations for how, based on results, women's and children's health could be further prioritized.Participating experts were informed of the intent and purpose of this research and were informed that their views would help shape the content of this policy analysis. They were made aware that the interviews would guide the direction of the document and would be referenced anonymously in the paper. Oral consent was sought during the interview and the research paper was circulated to interviewees to ensure that content reflected the discussions.This analysis has certain limitations. Because policies were identified using a health keyword search, a number of analysed policies may not have referred to health issues, i.e. policies nominating regional candidates for the position of Secretary General of the World Health Organization. As such the analysis of the 'other' policies that refer to health, mix policies on health issues, i.e. avian flu as well as policies on social determinants, as well as policies on political issues.Additionally, the low number of discussions with experts to inform policy implications and recommendations and the fact that interviewed experts are all current office bearers represents a limitation, as their expertise did not expand over the full period of existence of the Organization of African Unity and African Union.Since 1963, 55 African Union summits resulted in a total of 884 policies. 25% of these policies focus on peace and security, 17% on political affairs, and 14% on African Union governance. Economic and social affairs policies account for 11% and 12% of all AU policies respectively than do AIDS/TB/malaria-related policies (2.5 frames per policy). Within women's and children's health-related policies, human rights policies use the most frames to justify investments in women's and children's health and development. Overall, AIDS/TB/malaria policies use the health and economic frames slightly more often than women's and children's health policies Table .The frames used to prioritize women's and children's health based on our review of the 81 documents that mention health are now discussed in the following sections in alphabetical order.Women's and children's health is often framed using human rights, and the health of women and children is often addressed by rights policies. Between 2001 and 2010, there were 13 policies on women's and children's rights, empowerment and health, but there were only five policies specifically focused on Millennium Development Goals 4 and 5.Rights policies address health issues related to women and children and define government obligations to provide health services. For example, the 2005 Protocol to the African Charter of Human and People's Rights on the Rights of Women in Africa addresses an array of gender-related health issues and other determinants of health. These include: access to education and food; child labour and forced labour; physical and sexual violence; harmful traditional practices; early marriage; reproductive health rights; maternal mortality; cancers; menopause; and mental disorders.Legal, policy, and monitoring requirements-including human rights and African Union reporting-account for 42% of frames used by women's and children's health policies, 31% for AIDS/TB/malaria policies and 25% for other health policies.Advocacy statements, policies and related activities constitute 33% of the frames used in AIDS/TB/malaria policies, and 31% of those used in women's and children's health policies. Advocacy statements have considerably increased in policies that refer to women's and children's health.Referencing global goals and policies such as the Millennium Development Goals and regional events such as the World Summit on the Child has been significant. In the 2010 African Union Declaration on Maternal, Infant and Child Health and Development in Africa, member states:\"Individually and collectively reaffirm our previous commitments aimed at accelerating the health of our people and the social development of Africa. In this regard, we re-dedicate ourselves and commit our countries to accelerate efforts to improve the state of health of Africa's women and children and thereby attain all Millennium Development Goals (MDGs) particularly MDGs 4, 5 and 6 by 2015.\"Health frames include research, evidence and information on health outcomes and the burden of diseases. 17% of the frames used in AIDS/TB/malaria policies are health, 15% are in women's and children's health policies.Generally in these policies, claims of the severity of the women's and children's health situation are not substantiated by scientific data.Economic frames in policy include the use of arguments based on socioeconomic development, cost effectiveness and investment. Economic arguments constitute 13% of frames used in women's and children's health policies, 19% of those used in AIDS/TB/malaria policies and 37% of those used in other policies that mention health.\"Recognises with concern that poor maternal, infant and child health remains a major challenge confronting the continent and undermining its development efforts\".The use of the economic frame in women's and children's health policies is infrequent and unspecific. An example is seen in the 2008 Declaration on Maternal, Newborn and Child Health, which: \"African Women produce over 70% of the food crops in many parts of the Continent\".The bulk of policies that cite the contribution of women and children to development do not define or quantify it, or estimate the consequences of ill health. The closest numerical definition of the contribution of women to development is seen in the 1990 Resolution on the World Summit states that: The cost-effectiveness of women's and children's health interventions is also rarely used as an argument.The document review yielded some findings that can be used to shape knowledge and advocacy efforts to prioritize and strengthen references to women's and children's health in African Union Policies.The African Union predominantly views the health of women and children as a human right. This is seen in the inclusion of health entitlements in rights policies as well as in the referencing of rights policies in women's and children's health policies. Those organizations and individuals wishing to promote a post-2015 agenda for health can make use of the continued framing of women's and children's health in the rights discourse . The rig\"as the African Union becomes stronger, these policies will become more important because most of them incorporate monitoring and reporting clauses.\"Accountability and monitoring requirements are heavily referenced in African Union policies, and receive substantial attention within the assemblies. These requirements provide an entry point for the issue to remain on the policy agenda. This use of the accountability may become more pertinent, especially as the recognition of the African Union increases globally, regionally and nationally. One public health expert notes that Advocacy plays an important role in prioritizing issues. In part, this is due to the increased participation of advocates in African Union dialogue since the inception of the African Union in 2002. Advocacy for policy prioritization takes many shapes. For instance, prior to and during the July 2010 Summit, many articles appeared in Kampala and regional journals pointing to efforts undertaken by some countries to improve women's and children's health, highlighting the lack of progress in others and comparing regional health spending by all countries -28.While the use of the advocacy frame has been successful it has been challenging for the women's and children's health community. As noted by a public health specialist:Where the AIDS paradigm has a central message, and malaria has insecticide-treated nets, [advocacy for] reproductive, maternal, newborn and child health is complex. The messages range from coverage of different interventions along the continuum of care, to health systems issues and social determinants. When disaggregated, we note that a lot of them fall outside of the health sector. Grappling with the complex recommendations behind reproductive, maternal, newborn and child health has been challenging, and advocacy messages need to be simplified.\"\"The women's and children's health community should pursue its efforts in aligning and simplifying communication on key interventions to maximize campaign impact.While scientific evidence on trends in Millennium Development Goals 4 and 5 and related interventions, policies, financing and practices exists, Health advocates may choose to look at more effective ways to package, promote and disseminate these data to policy-makers in countries to ensure that policies are evidence-based. In addition to demonstrating that the implementation of affordable packages of interventions can save lives, they may also consider that this information will point to the severity of the women's and children's health crisis. As noted by one public health specialist:\"The AIDS and malaria movements, using health outcomes and impact of these diseases on the societies, have managed to position these diseases as crises. While stakeholders acknowledge that maternal, newborn and child health is a problem, they are rarely aware of the magnitude of this problem, and this is because [the health] community has not positioned the issue using scientific communication. We have tabled our messages on emotional argumentation.\"In addition to providing scientific evidence on the health burden, its causes and remedies, increased effort to demonstrate the economic impact of women's and children's health on national and regional economies would help to prioritize this issue among Heads of States. This is reflected in the intervention of certain Heads of States during the 2010 Summit who noted that while the health of women and children is a cause for concern, the attention provided to the issue must be shared with other sectors that have a direct bearing on countries' growth and productivity .Evidence of the productivity gains related to improved women's and children's health exists, though it is sparse and tends to reflect global figures. USAID estimates annual global losses in productivity due to maternal and newborn deaths at US$15 billion . RegionaThe proven cost-effectiveness of women's and children's health interventions is also rarely used as an argument, although clear evidence to support investment does exist. For example, research has indicated that for every dollar spent on family planning, four dollars are saved on treatment for complications of unwanted pregnancies .However, economic evidence varies greatly in quantity, quality and availability, which perhaps explains the restricted use of the economic frame. Traditional reluctance to justify health investment in economic terms may also be a factor. One health expert notes:\"There has traditionally been some resistance by the health community to using economic arguments to justify health investments because of a reluctance to associate health spending to returns to the state. This trend is slowly subsiding and research on the impact of health investment is increasing.\"Formal processes for prioritizing issues within African Union Assemblies include African Union Commission requests and recommendations; member state requests; and outcomes of regional ministerial conferences.\"One day when optimal processes have been put in place we will have reached global goals through continental mechanisms. CARMMA is one of these ideal processes.\"The African Union Commission aims to prioritize health issues based on member state interests and concerns . This gives programmes legitimacy and encourages their implementation by countries. One expert notes the importance of this consultative process in the success of health campaigns resulting from initiatives such as the International Conference for Population and Development (ICPD). Another expert states the need to take account of local factors when pursuing globally agreed goals: In addition to supporting the reflection of country priorities in the African Union agenda, the Commission plays a crucial role in defining it. The current Commissioner for Social Affairs has successfully positioned women's and children's health as a crucial issue for the continent, and has been the principal voice behind the regional campaign targeting its improvement. The Campaign for the Accelerated Reduction of Maternal Mortality in Africa (CARMMA) is an example of a successful campaign being pushed by the Commission. Having been launched at the highest political level in 34 countries in Africa (as of March 2011), CARMMA reflects successful national ownership and prioritization of women's and children's health .The Commission also monitors the implementation of policies. However, the proliferation of policies and the limited personnel and resources of the African Union Commission make the implementation of these policies and efforts to hold stakeholders accountable, daunting tasks. The monitoring of implementation in countries is a particular challenge. One expert notes:\"African Union Declarations provide ministries of health with the leverage we need in our national, regional and international advocacy. However more needs to be done to ensure strong, compelling monitoring of the implementation of these policies in countries. Partners need to support the African Union so that it can systematically provide sound progress reports on the implementation.\"Other regional institutions also contribute to prioritization of health on the continent. The Pan African Parliament, created in 2001, to ensure a representation of the voices of African populations in regional decision- making, intervened in favour of maternal, newborn and child health during the July 2010 Summit. While the body currently does not have legislative powers, it aspires \"to evolve into an institution with full legislative powers, whose members are elected by universal adult suffrage\" . An asseCurrently relegated to an advisory role, the body still contributes to regional prioritization. For instance, in October 2010, the Pan African Parliament Assembly adopted a motion adopting the Africa Parliamentary Policy and Budget Action Plan for Implementation of July 2010 AU Summit Decisions on Maternal, Newborn and Child Health and Development in Africa, and Partnership for Eradication of Mother to Child Transmission of HIV and AIDS . In 2011Similarly, the African Court on Human and People's Rights has a strong health element to its work . To dateThe Africa Peer Review Mechanism (APRM) assesses progress on health through its socioeconomic development remit . For exaStrong actor networks can generate, through advocacy efforts, increased attention for women's and children's health at both country and regional levels . The resFor example, during the Summit, health campaigners, development partners and Ministers of Health and Finance were invited to debate health financing during a high-level side event on health financing. Advocates were also invited to speak alongside Heads of States in the plenary session and requests from advocates such as Professor Jeffrey Sachs, Director of The Earth Institute at Columbia University, were included in the Summit outcome document.With advocacy playing such an important role, it is crucial to harness the efforts of women's and children's health advocates by developing multi-stakeholder actor networks that reflect the needs of member states, while also spanning the continuum of care for women's and children's health . Until 2This division in policy topics across the continuum of care has also been seen in the actor networks. The first streamlined global advocacy efforts for women's and children's health began as recently as 2005, when three global partnerships merged: the Partnership for Safe Motherhood and Newborn Health; the Healthy Newborn Partnership; and the Child Survival Partnership.Although major networks of this type do not yet exist at regional level, effective actor networks are emerging and contributing to priority setting and policy implementation. Civil society groups have long been recognized as key actors in advocacy, and the development of the Africa Maternal, Newborn and Child Health Coalition, a regional NGO network on women's and children's health, was an important outcome of the July 2010 Summit . One exp\"Coordinated partner action at the regional and country level is crucial to ensure the wide dissemination of Head of State commitments and accountability for results. NGO groups can be particularly helpful in this sense.\"Health-care professional associations in Africa can also influence priority setting and the accountability of states to produce results, either directly or by raising awareness of key issues ,49. ThesNon-traditional actors are beginning to play a role in advocacy for women's and children's health, including parliamentarians and the The [women's and children's health] community must involve non-traditional stakeholders in the reshaping of advocacy messages and widen their scope of engagement.\"\"To optimise their effectiveness nationally and regionally, actor networks should coordinate their work and involve the widest range of possible stakeholders. The Global Strategy for Women's and Children's Health provides some direction on expanding the types of stakeholders involved in women's and children's health efforts and notes the roles to be played by each of these different partner constituencies . The proThe Commission on Information and Accountability for Women's and Children's Health has also anchored its recommendations on principles of national sovereignty and has placed the country at the center of accountability processes. Its recommendations are based on principles of national leadership and ownership of results, strengthening countries' monitoring and evaluation capacity and reducing reporting burdens of countries .Funding and resources are also key factors, because women's and children's health initiatives can be hampered by the lack of a funding mechanism. One expert notes:\"Financing does impact policy prioritization ... in order to be most efficient and effective [funding] must come through a defined platform\" [latform\" .The benefits of sustained, coordinated funding are reflected in the prioritization of AIDS/TB/malaria in African Union policies. The Global Fund plays a major role here, contributing a high percentage of official development assistance and development assistance for health funding .Global actors have recognized the need for similar sustained commitments, and a substantial increase in health investment to close the \"funding gap\" for women's and children's health ,59. ReceIn 2007 and 2008, US$4.7 billion and US$5.4 billion were committed to women's and children's health activities in developing countries, respectively. These amounts reflect a 105% increase between 2003 and 2008 and improved prioritization of countries based on the burden of maternal and child deaths. However more remains to be done .Despite these new high-level commitments and subscription to the principles outlined in the Paris Declaration and the Accra Agenda for Action ,64, fundThis analysis demonstrates that in African Union policies, women's and children's health is prioritized using fewer frames than AIDS/TB/malaria. It also demonstrates that women's and children's health is most often framed using advocacy and accountability as opposed to health and economic arguments.Prioritizing women's and children's health in African Union policies could be strengthened by 1) widening the range of frames used to discuss these issues, and 2) developing a stronger evidence base to support both currently-used and new frames.The analysis also notes that regional institutions have and continue to play an important role in prioritizing women's and children's health. It identifies advocacy as an important mechanism to prioritize women's and children's health while ensuring links between global, regional and national priorities, and an alignment of partner efforts in reproductive, maternal, newborn and child health.In order to improve the prioritization of women's and children's health within African Union policy-making, women's and children's health stakeholders could: 1) support the African Union Commission and other regional bodies such as the Pan African Parliament in the prioritization of women's and children's health in policies, 2) strengthen existing regional and national women's and children's health networks in part by widening the range of partners to include the private sector, media and parliamentarians, and 3) align global efforts and campaigns with nationally-and regionally-defined priorities.In an environment where different priorities compete for funding, women's and children's health policies could claim more attention by using a wider range of frames. As noted by Heads of States in their Summit debate on women's and children's health, resource constraint is a key barrier to health investment. Therefore, the use of all available evidence would help advocates to make a more effective case for prioritizing women's and children's health in African Union policies.It seems likely that increased use of economic arguments would define women's and children's health as an investment, highlight the cost-effectiveness of health interventions and assist decision-makers in allocating scarce resources, by defining the synergies between investments in social determinants and health outcomes.To inform policy decisions more effectively, national and regional institutions have the option of gathering region-specific evidence on the cost savings that result from effective interventions, and on the impact of better health on growth and productivity. In the interim, existing global estimates, and regional calculations based on disability adjusted life years and gender disaggregated information, are available to inform policy discourse. Regional institutions could also gather evidence on the impact of investment on social determinants of health. This would inform debate by decision-makers about how to invest in other sectors that have an impact on women's and children's health, such as water and sanitation, education and transport ,68.The Lancet and in processes such as Countdown to 2015.Increased use of health frames would also aid the case for greater prioritization of women's and children's health, by providing compelling, specific evidence for the effectiveness of proven interventions. Much of the evidence on health outcomes already exists, and is reported by international journals such as The African Union Commission and related bodies such as the Pan African Parliament are key fora for advocacy in favour of women's and children's health. However, their resources are stretched, and they would benefit from increased capacity to engage fully in advocacy and follow-up at regional, sub-regional and national levels. Increasing the capacity of regional institutions can be done through the financing of health-related programs, specific projects and staff. Additionally, partners with wide country presence have the option of entering into joint planning with regional entities and acting as the implementing arms of regional agreements. For instance, partners could track a certain number of indicators to monitor progress on women's and children's health in countries which regional institutions can feed into their assemblies. These processes already exist but could benefit from being used more systematically and by a wider range of partners.While women's and children's health actors exist and are quite active, they have tended to operate in silos and focus on specific issues. We suggest that stronger and better coordinated actor networks, focusing across the continuum of care, would generate more effective advocacy, and encourage policy-making and implementation. National and regional committees on women's and children's health could be expanded to include-in addition to representatives from the Ministries of Health, donors and international organizations, health care professionals, academics and NGOs-the private sector, parliamentarians and the media. Collaboration among these partners would align efforts for more efficiency, improve the effectiveness of outlined strategies and widen the scope of actions to improve health.Women's and children's health actors also have the potential to facilitate better integration of national priorities in regional and global policies. For instance, the development of the Global Strategy sought to integrate perspectives from countries through consultations with country representatives. In the future, similar processes might use established regional networks such as the Africa Maternal, Newborn and Child Health NGO Network and processes such as African Union Ministerial meetings to ensure that global initiatives are based on national and regional priorities and recommendations.Campaigners for improved women's and children's health in Africa still face significant challenges. These include the limited capacity of the African Union bodies, the imperfect integration of national, regional and global priorities, and the lack of a central mechanism for the funding of women's and children's health. However, as our paper suggests, there is great scope for the African Union, other regional institutions and actor networks to work together more closely to develop and prioritize policies that improve the health of women and children in Africa.1. 1963 Health, Sanitation and Nutrition2. 1979 Declaration on the Rights and Welfare of the African Child3. 1985 Resolution on the Drawing up of a Programme of Assistance to Africa by UNESCO in the Fields of Scientific Research and Development4. 1987 Declaration of Health as a Foundation for Development5. 1987 Resolution on Universal Immunization in Africa6. 1987 Resolution on the Reconstruction of Chad7. 1987 Resolution on the Candidature of Professor Gottlieb Lobe Monessoko for the Post of Director General of WHO8. 1990 Declaration of the Assembly of Heads of States and Government of the Organization of African Unity on the Political and Socio-Economic Situation in Africa and the Fundamental Changes Taking Place in the World9. 1990 Resolution on the adoption of the African Charter on the rights and welfare of the African Child10. 1990 Resolution on the World Summit on Children11. 1991 Declaration of the Twenty Seventh Ordinary Session of the Assembly of Heads of States and Government on Employment in Africa12. 1991 Declaration on the current African Health Crisis13. 1992 Declaration on AIDS epidemic in Africa14. 1992 Resolution on AIDS and Africa: an agenda for action15. 1994 Tunis Declaration on AIDS and the Child in Africa16. 1994 Preamble to the Declarations and Resolutions adopted by the 30th Ordinary Session of the Assembly of Heads of State and Government17. 1995 Addis Ababa Declaration on the Dakar African Platform for Action on Women18. 1995 Declaration on the African Plan of Action concerning the situation on Women in Africa in the context of Family Health19. 1995 Resolution on Mobilization of Resources for Africa's Economic and Social Development20. 1996 Yaound\u00e9 Declaration on Polio Eradication in Africa21. 1996 Resolution on the Regular reporting of the Implementation status of OAU Declaration on HIV/AIDS22. 1996 Resolution on Bioethics23. 1997 Harare Declaration on Malaria Prevention and Control in the Context of African Economic Recovery and Development24. 1998 Ouagadougou declaration25. 1998 Decision: Malaria Prevention and Control within the context of Africa's Economic Recovery and Development26. 1999 Decision on the \"First Meeting of States Parties to the Convention on the Prohibition of the Use, Stockpiling, Production and Transfer of Anti-Personnel Mines and on their Destruction\"27. 2000 Lome Declaration28. 2000 Lome Declaration on HIV/AIDs in Africa29. 2000 Decision on proposal for the eradication of tsetse flies on the African continent30. 2000 CSSCDA Solemn Declaration31. 2001 Decision on the declaration of 2001-2010 as the decade for traditional medicine32. 2001 Decision on the report on implementation of the PoA on the eradication of tsetse flies in Africa.33. 2002 Decision on the control of Arterial Hypertension in Africa34. 2002 Decision on the status report on Global Alliance for Vaccines and Immunization (GAVI)35. 2003 Decision on promoting the development of sustainable cities and towns in Africa36. 2003 Declaration on the fifth WTO Ministerial Conference37. 2003 Maputo Declaration on Malaria, HIV/AIDS, Tuberculosis and Other Related Infectious Diseases (ORID)38. 2004 Decision on the Implementation of the New Partnership for Africa's Development (NEPAD)39. 2005 Decision on the Interim Report on HIV/AIDS, Tuberculosis, Malaria and Polio40. 2005 Decision on the proposal on sickle-cell anaemia41. 2005 Decision on the Report of the Commission on Accelerating Action for Child Survival and Development in Africa to meet the MDGs42. 2005 Declaration on the Review of the Millennium Declaration and the Millennium Development Goals (MDGs)43. 2006 Decision on the Linkage between Culture and Education44. 2007 Decision on avian flu45. 2007 Addis Ababa Declaration on Science Technology and Scientific Research for Development46. 2008 Decision on promotion of maternal, infant and child health and development47. 2008 Decision on the Report on the Promotion of Maternal, Infant and Child Health in Africa48. 2008 Decision on the Progress Report on the Implementation of the Commitments of the May 2006 Abuja Special Summit on HIV/AIDS, Tuberculosis and Malaria (ATM)49. 2009 Decision on the specialized technical committees50. 2009 Decision on the Report of the Implementation Status of Decision on Promotion of Maternal, Infant and Child Health and Development in Africa51. 2009 Decision on the Themes of the July 2009, January 2010 and July 2010 Sessions of the Assembly52. 2010 Declaration on information and communication technologies in Africa: Challenges and prospects for development53. 2010 Decision on the Five (5)-Year Review of the Abuja Call for Accelerated Action Towards Universal Access to HIV/AIDS, Tuberculosis and Malaria Services in Africa54. 2010 Declaration on Actions on Maternal, Newborn and Child Health and Development in Africa By 201555. 2010 Decision on the partnership for the eradication of mother to child transmission of HIV/AIDS56. 2010 Decision on the Report of Head of State and Government Orientation Committee on NEPAD1. 1992 Resolution on the Summit on the Economic Promotion of rural women presented by Senegal2. 1997 Decision: Harare Declaration on Malaria Prevention and Control3. 1998 Establishment of an African Fund for AIDS Control4. 1999 Decision on the ILO Convention on the Banning of the Worst Forms of Child Labour and Immediate Action for their Elimination5. 2000 Decision on the Report of African Summit on Roll-Back Malaria6. 2000 Decision on the Holding of an African Summit on HIV/AIDS, Tuberculosis and other related infectious diseases7. 2001 Decision on the African Summit on HIV/AIDS, Tuberculosis and other related infectious diseases8. 2001 Decision on the Pan-African Forum on the future of Children9. 2002 Decision on the report of the African Committee on the rights and welfare of the Child10. 2003 Decision on the Appointment of Members of the African Commission on human and people's rights11. 2003 Decision on the Draft Protocol to the African Charter on Human and People's Rights Relating to the Rights of Women12. 2004 Decision on AIDS Watch Africa (AWA) and the Implementation of the Abuja and Maputo Declarations on Malaria, HIV/AIDS, Tuberculosis and Other Related Infectious Diseases in Africa13. 2004 Decision on the International Centre for the Education of Girls and Women in Africa (CIEFFA)-Doc. Assembly/AU/11 (V) Add.114. 2004 Solemn Declaration on Gender Equality in Africa15. 2005 Decision of the appointment of members of the African experts on the rights and welfare of the child16. 2006 Decision on the Progress Report on AIDS Watch Africa (AWA)17. 2006 Decision on the election of one member of the African committee on the rights and welfare of the child18. 2006 Decision on Abuja Call for Accelerated Action Towards Universal Access to HIV/AIDS, Tuberculosis and Malaria (ATM) Services in Africa19. 2006 Decision on the continental framework for harmonization of approaches among member states and integration of policies on human rights and people affected by HIV/AIDS in Africa20. 2007 Decision of the reports on the implementation of the African Union solemn declaration on gender equality in Africa21. 2008 Decision of the reports on the implementation of the African Union solemn declaration on gender equality in Africa22. 2008 Decision of the appointment of members of the African experts on the rights and welfare of the child23. 2009 Decision on the African women's decade24. 2010 Decision on the Establishment of the Fund for African Women25. 2010 Decision on the Appointment of Members of the African Committee of Experts on the Rights and Welfare of the ChildThe interviewees were asked to comment on:\u2022 The prioritization of women's and children's health in the African Union; whether this could be improved;\u2022 Which mechanisms and actors have contributed to the prioritization or de-prioritization of women's and children's health; and how these same mechanisms and actors could improve prioritization.\u2022 Their perception of the value placed on African Union policies by states and development partners, and how this value could be increased.APRM: Africa Peer Review Mechanism; AU: African Union; AWA: AIDS Watch Africa; CARMMA: Campaign for the Accelerated Reduction of Maternal Mortality in Africa; EU: European Union; GAVI: Global Alliance for Vaccines and Immunization; ICPD: International Conference for Population and Development; MDGs: Millennium Development Goals; OAU: Organization of African Unity; PMNCH: Partnership for Maternal, Newborn & Child Health; WAHO: West African Health Organisation.The authors declare that they have no competing interests.KT: Conducted research and coding and wrote different iterations of the research article. RS: Advised on content of the research and assisted in conducting key expert interviews. SK: Contributed to the analysis and writing process and content. ES: Coded policies using the coding framework and helped refine coding framework. FB: Contributed to the design, analysis and writing of the paper. BO: Guided development of the overall paper. All authors read and approved the final manuscript."} +{"text": "There were errors in both the Funding and Competing interests.The correct versions of both are:Funding: This study was supported by the BIDMC Department of Medicine Foundation and Global Research Laboratory Award . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Competing interests: The authors have read the journal's policy and have the following conflicts: CJL has a management position at Boston Biomedical, Inc, a company that develops cancer stemness inhibitors. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials."} +{"text": "The Journal of Spinal Cord Medicine's third with our new international publisher, Maney Publishing. I am pleased to report that the feedback on JSCM's new look has been overwhelmingly positive. Of greater importance is that with Maney's expanded online article access, the journal is more widely available to professionals and libraries around the world.This issue marks JSCM as a benefit for their members\u2014the Academy of Spinal Cord Injury Professionals (the Academy) and the American Spinal Injury Association (ASIA).For the journal and the dynamic field of spinal cord injury, there's never been a more exciting time. The broad range of research and clinical activities of interest to our readers is evident in the 2011 meeting agendas for the two organizations that feature ISCoS-ASIA 2011 International Conference on Spinal Cord Medicine and Rehabilitation. More than 750 scientists and clinicians will be in attendance for the joint conference program, as well as the Spine Symposium on June 4 and the pre-course, The State of the Science in Spinal Cord Injury Rehabilitation, beginning June 5.From June 4 to 8, ASIA joins with the International Spinal Cord Society (ISCoS) for their shared meeting in Washington, DC\u2014SCI Medicine - An Intensive Review Preconference Workshop. This comprehensive review of assessment and treatment and rehabilitation strategies is highly recommended, especially for those preparing for the Subspecialty SCI Certification and Maintenance of Certification Examination offered by the American Board of Physical Medicine & Rehabilitation.Las Vegas, Nevada is once again the site for the Academy's annual conference, this time hosted at Rio All Suites Resorts and Casino from September 5-7. Look for the return appearance of Dr. Vernon Lin in JSCM looks forward to bringing you interdisciplinary content reflecting the diverse topics addressed in Washington, DC, and Las Vegas.Make time in your schedule for these terrific opportunities for education, inspiration, networking and relaxation. Comments and suggestions from our readers are always welcome."} +{"text": "There is an error in the funding statement. The correct funding is, \u201cThis work was conducted with support from Harvard Catalyst / The Harvard Clinical and Translational Science Center . The content is solely the responsibility of the authors and does not necessarily represent the official views of Harvard Catalyst, Harvard University and its affiliated academic health care centers, the National Center for Research Resources, or the National Institutes of Health. In addition, the funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "There was an error in the Funding statement. The correct Funding statement is available below:This work was supported by Guangdong Province Talent Introduction Project Grant and National Natural Science Foundation of China grant (81141115) to MH. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The following information is missing from the funding statement. \"This work was supported by the Calleva Research Centre for Evolution and Human Sciences (DB).\"The full funding statement is, \"AM and MNA are funded by UT and IPM. BB was supported by a postdoctoral fellowship from the British Academy and European Research Council (Grant number: 309865). This work was supported by a StG grant from European Research Council to BB. This work was supported by the Calleva Research Centre for Evolution and Human Sciences (DB). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "The funding project numbers were omitted from the published Funding statement. The correctFunding is:This work was supported by the grants from the Ministry of Science and Technology of China. The funders had norole in study design, data collection and analysis, desicion to publish, or preparation ofthe manuscript."} +{"text": "There was an error in the financial disclosure. The correct financial disclosure statement is: \"This work was supported by grant 2/0104/09 from Scientific Grant Agency of the Ministry of Education of the Slovak Republic and the Slovak Academy of Sciences. MDH is supported by a Biotechnology and Biological Sciences Research Council Studentship. JCC was supported by Biotechnology and Biological Sciences Research Council grant BB/G019347/1 to M.J.Butner (John Innes Centre) and N.Le Brun (University of East Anglia). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There is an error in the Funding section. The correct funding information is as follows: The work was supported by the National Natural Science Foundation of China (NSFC) (No. 81101782), NSF project CQ CSTC (No. CSTC2011BB5020), and NSF Third Military Medical University (No. 2010XQN36).Also, it should be noted that the date found in the footer of the downloadable PDF version of this article is incorrect. The correct date of publication for this article is April 2012, as stated in the Abstract and citation."} +{"text": "The correct version of the Funding statement should read \"The Special Research Fund (BOF) partly funded this research. The University Foundation of Belgium also provided funding for this research. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "Sir, We thank Dr Vishnu for his interest in our paper. We reported atrophy of the caudate and thalamus in presymptomatic familial Alzheimer\u2019s disease mutation carriers at a stage when hippocampal atrophy was not yet evident . It was undertaken at UCLH/UCL, who received a proportion of funding from the Department of Health's National Institute for Health Research (NIHR) Biomedical Research Centres funding scheme, and was supported by the NIHR Queen Square Dementia Biomedical Research Unit. NCF is a NIHR senior investigator. The Dementia Research Centre is an Alzheimer's Research UK Co-ordinating Centre and has received equipment funded by Alzheimer's Research UK and Brain Research Trust.This work was supported by the"} +{"text": "There was an error in the published Funding statement. The correct Funding is: This study was supported by the Swiss National Science Foundation (Grant n\u00ba 32003B_127619) and by funds from the Department of Intensive Care Medicine, University Hospital and University of Bern. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Following publication of our article , we becaThis project was funded under Contract No. HHSA290200710062I from the Agency for Healthcare Research and Quality, U.S. Department of Health and Human Services. The authors of this report are responsible for its content. Statements in the report should not be construed as endorsement by the Agency for Healthcare Research and Quality or the U.S. Department of Health and Human Services."} +{"text": "There were errors in the Funding statement and in Table 3.The correct Funding statement is: This study was funded by National Natural Science Foundation of China (Grant No.31270496). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.A correct version of Table 3 is available here:"} +{"text": "Multiple funding organizations and grants were incorrectly omitted from the Funding Statement. The Funding Statement should read: \"This work was supported by the Beijing Natural Science Foundation , the Beijing Excellent Talent Project (project number PHR20100510), Tianjin Health Bureau Technology Fund and the Beijing Key Laboratory of Major Brain Disorders (project number 2011NZDB01). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There is an error in the funding statement. The correct funding statement is, \"Project supported by United States National Institutes of Health (NIH) Grants DA013261 DA019676 and MH083840. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There was an error in the Funding statement. The correct funding statement is: This work was supported by a National Research Foundation of Korea (NRF) grant (No. 2010-0014269), Republic of Korea. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Host and microbial molecular dissection of pathogenesis and immunity in tuberculosis; Grant agreement no.: 200732) to LG; by Italian Association for Research against Cancer (AIRC) grants to LG and FB. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\" A funding organization for the 12th and 14th authors was incorrectly omitted from the Funding Statement. The Funding Statement should read: \"This work was funded by the European Community Seventh Framework Programme (Project acronym: HOMITB, Title:"} +{"text": "A funding organization was incorrectly given in the Funding Statement. The Funding Statement should read: \"Results for Development's Center for Health Market Innovations provided funds to complete this systematic review. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "The correct Funding statement is, \"This work was partly supported by the special fund for public scientific research from the Ministry of Water Resources of China (NO.201001003)"} +{"text": "Life expectancy has been increasing during the last century within the European Union (EU). To measure progress in population health it is no longer sufficient to focus on the duration of life but quality of life should be considered. Healthy Life Years (HLY) allow estimating the quality of the remaining years that a person is expected to live, in terms of being free of long-standing activity limitation. The Joint Action on Healthy Life Years (JA: EHLEIS) is a joint action of European Member States (MS) and the European Union aiming at analysing trends, patterns and differences in HLY, as well as in other Summary Measures of Population Health (SMPH) indicators, across the European member states.The JA: EHLEIS consolidates existing information on life and health expectancy by maximising the European comparability; by analysing trends in HLY within the EU; by analysing the evolution of the differences in HLY between Member States; and by identifying both macro-level as micro-level determinants of the inequalities in HLY. The JA: EHLEIS works in collaboration with the USA, Japan and OECD on the development of new SMPHs to be used globally. To strengthen the utility of the HLY for policy-making, annual meetings with policy-makers are planned.The information system allows the estimation of a set of health indicators for Europe, Member States and shortly their regional levels. An annual country report on HLY in the national languages is available. The JA: EHLEIS is developing statistical attribution and decomposition tools which will be helpful to determine the impact of specific diseases, life styles or other determinants on differences in HLY. Through a set of international workshops the JA: EHLEIS aims to develop a blueprint for an international harmonized Summary Measure of Population Health.The JA: EHLEIS objectives are to monitor progress towards the headline target of the Europe 2020 strategy of increasing HLY by 2 years by 2020 and to support policy development by identifying the main determinants of active and healthy ageing in Europe. Life expectancy at birth has steadily increased during the last decade in the EU27, by more than 3 years for men and 2 years for women, leading to accelerated population ageing. However gaps among Member States of the European Union (EU) have remained large, 13 years in both 1997 and 2010 for men, although for women gaps have slightly reduced from more than 9 years in 1997 to 8 years in 2010. Additionally the gender gap in life expectancy at birth remains high at 6 years in 2010 although it also has reduced over time. When the quality of the years lived is taken into account through health expectancies such as life expectancy in good perceived health, life expectancy without chronic morbidity, or life expectancy without disability, even larger gaps are seen among European countries. For instance, life expectancy without long-standing activity limitations or Healthy Life Years (HLY) show gaps in both men and women reaching close to 20 years in 2010 among Member States[The HLY is technically a Summary Measure of Population Health and part of the family of disability-free life expectancy indicators which measure the number of remaining years that a person of a certain age is expected to live without disability. A Summary Measure of Population Health brings together data on both the length and the quality of life and are As measures of population health, HLY and health expectancies in general have several strengths. Health expectancy indicators are more intuitive and understandable for policy makers and the public at large compared to mortality rates or morbidity incidence or prevalence rates. Health expectancies are typically estimated based on cross-sectional survey data and guidThe next step in this process was to structure and harmonize the calculation, analysis and interpretation of data collected through those surveys. This goal was achieved by the European Health Expectancy Monitoring Unit (EHEMU) and the European Health and Life Expectancy Information System (EHLEIS) project. EHEMU established an information system which facilitates the calculation of life and health expectancies in the EU and its Member States. This information system provides a central facility for the co-ordinated analysis and synthesis of life and health expectancies and provides evidence of inequalities between Member States, highlighting potential targets for public health strategies both nationally and at a pan-European level. EHLEIS extended the EHEMU activities by more in-depth analysis of trends and gender gaps in health expectancies,14 and oMaintain and continue developing a European Health and Life Expectancy Information System, in order to improve and harmonise calculations for the development of the Healthy Life Years \u2013 HLY \u2013 structural indicator\u201d. Accordingly the JA: EHLEISWithin the programme of the Community action in the field of health, JA: EHL(i) consolidates the existing information system (EHLEIS) through promoting further European comparability, annual HLY calculations and dissemination and upd(ii) further extends EHLEIS through analyses of micro-level health determinants using the new European Health Interview Survey (EHIS) and analyses of HLY gaps between socio-economic groups,(iii) develops an alternative Summary Measure of Population Health that can also serve as a common international measure beyond the EU,(iv) Integrates the Task Force on Health Expectancies into an In particular, in the trend and gap analyses the JA: EHLEIS utilises a range of health expectancies, including self-rated health, morbidity and disability. The JA also develops a study design for computing maximally comparable health expectancies by socio-economic status for the Member States that already have mortality data by socio-economic position. Finally, in collaboration with the main OECD countries, especially the United States of America (USA) and Japan, the JA: EHLEIS will provide a blue print for an improved new internationally comparable Summary Measure of Population Health, thus upgrading the current European HLY by a global indicator. A Summary Measure of Population Health that is comparable to the USA and Japan indicators is a necessary condition to achieve a grade \u201cA\u201d for European Structural Indicators. The international discussion will not focus on the health expectancy indicator as such but rather on the essential of how to measure the health or quality of life related information necessary to calculate the health expectancy.The Lancet in 2008 is a good example of HLY use in non-health policies[The JA: EHLEIS targets Member States, health and non-health policy makers at the regional, national and European level, health professionals and researchers, media, general public and Non Governmental Organizations (NGO). Increasing involvement of Member States in health monitoring is one of the main priorities of JA: EHLEIS as well as promoting their wider engagement in using HLY in policy-making. The annual public JA: EHLEIS meeting plays a central role in addressing these priorities. In addition, the work published by the EHLEIS group in policies. Scientipolicies,5 and onpolicies,16,20,21policies and Wikipolicies. The JA:A first major aim of the Joint Action is to maintain and continue developing the European Health and Life Expectancy Information System (EHLEIS), by providing online calculations of various health indicators with confidence intervals, that are comparable among EU countries, and with improved dissemination, understanding, ease of access and public health relevance. The methods and means to meet this aim include: computational and web methodologies for the EHLEIS database; statistical and demographic analyses for the substantive results; linguistic methods for the translation/provision of the easy-to-use web facilities in multiple European languages and translation of the Country reports into national languages; and methodologies to address the conceptual basis of the HLY indicator and its greater comparability with Summary Measures of Population Health in the USA and Japan. The JA: EHLEIS further develops the EHLEIS system to allow rapid access to up-to-date data on health expectancies from a variety of European databases , EHIS). The computational and web expertise to undertake this key foundation of the Joint Action come from Montpellier RIO Imaging. AnalysiThe main outcomes of the Joint Action will comprise:(i) An Information System allowing online calculation of health indicators ), combining population and mortality data with health information coming from European and/or national surveys. All HLY-related websites will be organized in a new website;(ii) Country reports on HE: Part of a series, each annual report presents life expectancy and HLY for the country of interest and for the overall EU27 using the SILC question on long term activity limitation (GALI) included in the SILC survey since 2004 or 2005. The reports will be translated in the national languages and will be disseminated to national institutions;(iii) Proceedings of the JA: EHLEIS annual meetings: The annual meetings will consist of one half-day open to the public and media (back to back with the Enlarged Steering Committee). The main aim of the meeting is to encourage Member States to use health expectancies, including HLY, in their health, social and employment policies;(iv) Presentation of the new HLY values and latest trends in Europe and press communiqu\u00e9s;(v) Statistical tools - New version of attribution and decomposition tools, distributed with updated manuals through the website and deve(vi) Technical reports and scientific analyses exploring geographical variations in HLY within Europe, trends in HEs in Europe, social differentials in HLY across Member States and calibration of the Global Activity Limitation Indicator (GALI) with EHIS data;(vii) A blueprint for a new international Summary Measure of Population Health.In addition the Joint Action plans to present its work at major European Conferences such as the European Health Forum in Gastein and the The JA: EHLEIS contributes directly to two of the three objectives of the Second Programme of Community Action in the Field of Health 2008\u20132013: to promote health and to generate and disseminate health information and knowledge. The JA:The authors declare that they have no competing interests.JMR and CJ drafted the manuscript. All authors commented the draft versions. All authors read and approved the final manuscript.http://www.eurohex.eu/index.php?option=aboutehemu#team.The JA: EHLEIS team:"} +{"text": "In the Funding statement, the number of a grant from the National Institutes of Heatlh to the first author (JK) is incorrect. The Funding statement should read: \"This research was supported by National Institutes of Health . The Pennington Biomedical Genomics Core Facility is supported in part by COBRE (NIH P20 GM103528) and NORC (NIH 2P30DK072476) center grants from the National Institutes of Health. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There were multiple errors and omissions in the Funding Statement. The Funding Statement should read: \"This work was supported in part by research grants of the Research Foundation Flanders , the Belgian Foundation against Cancer , the Methusalem program of the Flemish Government attributed to Prof Herman Goossens , the Interuniversity Attraction Pole program (IAP #P6/41) of the Belgian Government and the Belgian Hercules Foundation (www.herculesstichting.be). SA is a former PhD fellow of the Research Foundation Flanders and currently holds an Emmanuel van der Schueren Fellowship of the Flemish League against Cancer . SA was awarded a 2012 Endeavour Research Fellowship by the Department of Industry, Innovation, Science, Research and Tertiary Education of the Australian Government (www.innovation.gov.au). SA also received financial support from the Belgian Foundation against Cancer and the Belgian public utility foundation VOCATIO (www.vocatio.be). EL was supported by a PhD grant of the Institute for the Promotion of Innovation through Science and Technology and by an Emmanuel van der Schueren Fellowship of the Flemish League against Cancer. JT was supported by a grant from The Netherlands Organization for Scientific Research . ELS is a post-doctoral fellow of the Research Foundation Flanders. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.\""} +{"text": "The corresponding author of the article was not correctly indicated. The correct corresponding author is not the last author, but instead the second author, Lei Ren. The corresponding author's email address is: lei.ren@manchester.ac.uk.In addition, two grant numbers from the Key Project of the National Science Foundation of China were incorrectly listed in the Funding Statement. The Funding Statement should read: \"This work was supported by the International Cooperation Project of National Natural Science Foundation of China (No. 50920105504), a Biotechnology and Biological Sciences Research Council Grant (No.BB/H002782/1), the Project of National Natural Science Foundation of China (No. 51105167), the Fundamental Research Founds of China (No. Z201105), the Key project of National Science Foundation of China , the postdoctoral project and the scientific and technological development planning project of Jilin Province . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There is an error in the Funding statement that was omitted from the first correction. The complete, correct Funding statement is: The present study was supported by the National Natural Science Foundation of China , Scientific Research Foundation of Hangzhou Normal University in China (2011QPL15 to F. L.), and National Institutes of Health Grants . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There were errors in the Funding section. The correct funding information is as follows: This research was supported in part by facilitating funds from the George Washington University School of Medicine (SAM), National Science Foundation grant MCB-1121711 (SAM), and the Intramural Research Program of the National Institutes of Health, National Cancer Institute project number ZIA BC01000617 (IOD). Imaging was supported by NIH NICHD 2P30 HD040677 and NIH NCRR 1S10RR025565-01. Dr Klein\u2019s efforts were supported by the National Science Foundation while working at the Foundation. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the National Science Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The authors are grateful for a grant provided to CHC from Consejo Nacional de Ciencia y Tecnolog\u00eda (Scholarship No. 95802). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. There was an error in the funding statement. The correct funding statement is: The research was supported by the National Science Foundation (Grant no. 0347960 to LRG). MGS was funded in part by the European Community's Seventh Framework Programme (FP7/2007-2013;"} +{"text": "There was an error in the Funding statement.The correct version of the Funding is: This study was supported by the National Natural Science Foundation of China and Nature Science Foundation of Shaanxi Province (No. 2007K09-05(7)). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "A funding organization and grant were incorrectly omitted from the Funding Statement. The Funding Statement should read: \"The authors wish to thank the National Natural Science Foundation of China (41373073) and Zhejiang Science and Technology Program (2011F20025) for providing the financial support for this project. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. \""} +{"text": "Of all the goals listed in last year's President's address, most of them were realized. The National Health Personnel Licensing Examination Board (NHPLEB) of the Republic of Korea earned the ISO 9001 certificate in August 2011. The Journal of Educational Evaluation for Health Professions (JEEHP) was accepted as an official journal of the Association for Medical Education in the Western Pacific Region. This means that a new international window was opened for communicating ideas on educational evaluation within the Western Pacific Region. Another remarkable project was two trial runs of an examination of ubiquitous-based testing (UBT) for medical technologist students in November and December 2011. Smart pads were kindly provided by Korea Telecom. Some bandwidth problems and procedural mistakes in the first examination were corrected in the second one. The response from examinees and the faculty was favorable to UBT suggesting the possibility of introducing UBT to the variety of national health personnel licensing examinations. At last November's meeting of the Board of the Korean Medical Licensing Examination, the NHPLEB was also asked to prepare for the introduction of computer-based testing (CBT) within 7 years. 2012 is the 20th anniversary of NHPLEB. To continue the success of the last 20 years of hard work and perspiration, a conference will be held this May. Here, the new short-term and long-term goals of NHPLEB and new educational evaluation projects will be discussed in depth, preparing plans for the next 20 years worthy of our pride and admiration. Among those planned projects, UBT and CBT will be implemented to two health personnel fields, medical technologists and dentists as trial run of examination.I have been pleased to observe the increase in the papers published through this journal. In 2012, I hope an even greater number of invaluable articles will appear in the journal to promote higher quality educational evaluation for health personnel."} +{"text": "The following information is missing from the Funding section: The funding source for this project is internal to Mayo Clinic and was provided by the Center for Individualized Medicine at Mayo Clinic. No additional external funding was received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "A funding organization and grant were incorrectly included in the Funding Statement. The Funding Statement should read: \"This work was supported by the National Natural Science Foundation of China and Jiangsu Province's Outstanding Leader Program of Traditional Chinese Medicine. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There was an error in the Funding statement.The correct version of the statement is: This study was supported by a Plastic Surgery Education Foundation Research Fellowship: \u201cMechanisms regulating fibrosis in chronic lymphedema\u201d . Research reported in this publication was supported by the National Heart, Lung, And Blood Institute of the National Institutes of Health under Award Number R01HL111130. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The affiliation for the second author is incorrect. Carla Bosia is not affiliated with Department of Physics and INFN, University of Torino, Torino, Italy but with Human Genetics Foundation (HuGeF), Torino, Italy. The correct affiliation is listed as #2 on the PDF but is not present on the HTML version of the paper."} +{"text": "The Funding statement in the article was incorrect. The correct version of the Funding statement is available below.The authors acknowledge the following sources of funding: The University of Liverpool, UK provided research costs; The University of Guadalajara, Mexico provided research costs and the Peter Lienhardt Memorial Fund and Philip Bagby Bequest from the University of Oxford, UK provided a travel grant. The funders played no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The number of one of the grant from the first funding body, National Natural Science Foundation of China, is incorrect. The correct grant numbers from this funding body are: 30830072, 31271703."} +{"text": "There were errors in the Funding section. The correction funding information is as follows: S.S. gratefully acknowledges the support of the Josef Sagol Fellowship for brain research at Tel Aviv University. This research was supported by the I-CORE Program of the Planning and Budgeting Committee and The Israel Science Foundation (grant No 41/11). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There were errors in the Funding section. The correct funding information is: This research was supported by the Key Project for Breeding Genetic Modified Organisms (2011ZX08012-004). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The recipient of the grant from the NIH was incorrectly omitted from the Funding Statement. The Funding Statement should read: \"This research was supported by an NIH grant to Dr. Zili Zhang (R01 EY022937). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "In addition, the seventh author, Han Lin, should be indicated as a corresponding author. Han Lin's email address for the purposes of correspondence regarding the article is: In the Funding statement, the name of a funding organization is incorrect. The Funding statement should read: \"The work was supported in part by National Science Foundation of China (81271469) and Research Foundation of Doctor Program (20113321110003) to QL, Natural Science Foundation of Zhejiang Province (Z2101211) to QL. The funder is the corresponding author of this study, and he gave final approval of the version to be published.\""} +{"text": "There is a sentence missing from the Funding Statement. The following is the correct Funding Statement:This work was supported with funding from the National Science Foundation (DEB 0083944) and the U.S. Geological Survey Wildlife and Invasive Species Programs. The Department of Forest and Wildlife Ecology at the University of Wisconsin provided funds for publication. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "A funding organization for the 13th author, Richard Hamill, was incorrectly omitted from the Funding Statement. The following sentence should be included with the Funding Statement: \"RJH was supported in part by resources provided by the Department of Veterans Affairs.\"In addition, an affiliation for the 13th author was incorrectly omitted from the byline. Richard Hamill is also affiliated with The Michael E. DeBakey VA Medical Center, Houston, Texas, United States of America."} +{"text": "There was an error in the Funding statement. The correct Funding is:This work was funded by the Howard Hughes Medical Institute and also supported by National Institutes of Health grant CA095389 to IG. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The Funding statement is incorrect. The publisher apologizes for the error. Please view the correct funding information below:\"This work was supported by CONICYT \"Proyecto Anillo en Complejidad Social\" SOC1101 and by the Millenium Center for the Neuroscience of Memory, Chile (NC10-001-F), which is developed with funds from the Innovation for Competitivity from the Ministry for Economics, Fomentation and Tourism, Chile. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "Two funding organizations are incorrectly omitted from the Funding Statement. The Funding Statement should read: \"Funding was provided by the Australian Institute of Marine Science, the Australian Government\u2019s National Environmental Research Program and an Australian Research Council Grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "There was an error in the Funding statement. The correct version of the statement is available below.This work was funded by NIH R01 CA102184 (M.M). This work was also funded in part by the Intramural Research Program of the National Institute of Environmental Health Sciences-National Institutes of Health (projects: Z01ES100475 and Z01ES065079). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There was an error in the Funding statement. The correct statement should be:This work was supported in part by research grants from the Natural Science Foundation of China (#30772548 to Dr. Zhou), and Foundation from Excellent Master Dissertation of College of Laboratory Medicine at Chongqing Medical University (#201202). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "PROSPERO is an international database of prospectively registered systematic reviews in health and social care. Between July 2012 and June 2013, 1,106 registrations were added, bringing the total since launch in February 2011 to 1,704. The value of the growing number of records is reflected in a 117% increase in page views in the first half of 2013 compared with the first half of 2012. Developments over the year included expansion of scope, improvement of the registration form and easier access to information on how to register. PROSPERO was launched in February 2011 as the first open access international prospective register for systematic review protocols. The aim of the register is to help prevent unintended duplication of systematic reviews by allowing those commissioning or planning reviews to identify whether there are any relevant reviews already underway. PROSPERO also aims to provide transparency in the review process and help identify potential biases by enabling comparison of reported review findings with what was planned in the protocol. Registration, which is free, requires entry of a minimum dataset that includes key information about the systematic review design and brief administrative details and was developed and agreed through international consultation. This letter briefly outlines activity and progress of PROSPERO from 1 July 2012 to 30 June 2013.Since its inception, there is evidence that PROSPERO is gaining increasing acceptance and visibility. Numbers of protocol registrations and register users have increased rapidly; 1,106 records were added to PROSPERO between July 2012 and June 2013, bringing the total number of registrations to 1,704 over the past year. As IP addresses can represent either a single user or a whole organisation , we know that this is a conservative estimate of usage. Nevertheless, register usage greatly increased in 2013 with 117% more page views in the first half of 2013 compared with the same period in 2012. This reflected the growing number of searchable records in the register.In January 2013, we published data on the utility of PROSPERO in its first year [Presentations about PROSPERO were well received at the EQUATOR Scientific Symposium in Freiburg and the Plans for further PROSPERO developments include implementation of a new administration system with such functions as automated email reminders to help registrants keep their records up to date. The system will also facilitate the efficient addition of new Cochrane protocols to the register. A new search interface for the website is currently under development and is being informed by feedback from a recent survey of PROSPERO registrants and users.The PROSPERO team would like to thank the advisory group for their continued advice and support, and all registrants and register users. We always welcome feedback and would like to hear more from users, particularly if and how the register has been useful.CRD: Centre for reviews and dissemination; IP: Internet provider; NIHR: National Institute for Health Research.The development and ongoing management of PROSPERO is supported by the Centre for Reviews and Dissemination\u2019s (CRD) core work programme, which is funded by the National Institute for Health Research, England; the Department of Health, Public Health Agency, Northern Ireland and the National Institute for Social Care and Health Research, Wales.AB is a research fellow at NIHR CRD, University of York, UK, and is responsible for the development and maintenance of PROSPERO."} +{"text": "Some information was missing from the funding section. The correct funding statement should read: This study was supported in part by grant AI28900 from the National Institutes of Health, grant 3401-11 from the Leukemia and Lymphoma Society and grant 1024929 from the National Health and Medical Research Council. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Both Daniel Gough and David Levy are corresponding authors. Dr. Gough can be reached here : daniel.gough@monash.edu"} +{"text": "There was an omission in the funding information. The correct funding information is as follows: \"The study was sponsored by the National Research Foundation of Singapore , the Pharmacogenomics of Anticancer Agents Research (PAAR) Group, NIH/NIGMS U01GM61393 and the National Medical Research Council of Singapore (NMRC/CSA/021/2010). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There was an error in the published Funding statement. The correct Funding is:http://www.gacr.cz/); the Institute of Organic Chemistry and Biochemistry ; the Czech University of Life Sciences and the Ministry of Agriculture of the Czech Republic ; Japan Society for Promotion of Science ; Grants-in-Aid for Young Scientists, the Ministry of Education, Culture, Sports, Science and Technology of Japan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Financial support for this study was received from the Czech Science Foundation (project P506/10/1570;"} +{"text": "There was an error in the Funding statement. The correct version of the Funding statement is available below.The work was supported by NASA grant NNX10AJ31G and the LLUMC Department of Radiation Medicine. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There were errors in the Funding section. The correct funding information is as follows: This work was supported by Grant-in-Aid for Young Scientists (B) 23700476 from the Japan Society for the Promotion of Science, a grant from Naito Foundation and the Suzuken Memorial Foundation to SK, and grants from Dystonia Medical Research Foundation, Edward Mallinckrodt, Jr. Foundation, National Science Foundation and Whitehall Foundation to NCH. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. No additional external funding was received for this study."} +{"text": "The GEO study accession number was not noted in the published article. The GEO study accession number is GSE34197.There was an error in the funding statement. The correct funding statement is: This work was funded by National Institutes of Health grants GM083337 to DMG and NS048859 to ME, and a grant from the FSHD Global Research Foundation to ME. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The following information was missing from the Funding section: This work was supported by the NIH, National Institute of General Medical Sciences, Protein Structure Initiative [U54 GM094586 and GM074898]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In the published article the part of the funding statement describing how Philip Bejon was supported should read: \"PB is supported by the NIHR Biomedical Research Centre in Oxford.\" The corrected funding statement for this article reads in full: “The study was supported by the Wellcome Trust and by the Kenya Medical Research Institute (KEMRI). PB is supported by the NIHR Biomedical Research Centre in Oxford. KM, TNW, and FO are supported by the Wellcome Trust. SIH is funded by a Senior Research Fellowship from the Wellcome Trust (#079091). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.”"} +{"text": "There was an error in the published funding statement. The correct funding is:This study was supported by National Natural Science Foundation of China , and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Professor Hajo Grundmann was not included in the author byline. Prof. Grundmann should be listed as the sixth author and affiliated with Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands. The contributions of this author are as follows: Conceived and designed the experiments, performed the experiments, analyzed the data, and wrote the manuscript.In addition, there was an error in the Acknowledgments Statement. The Acknowledgments Statement should read: \"The authors wish to thank the Thika Hospital staff and patients for their involvement in surveillance and the SSI Surveillance team for their diligent work on data collection and entry. We also wish to thank Andy Hall and Neal Alexander for their assistance with designing and conducting SSI surveillance and Peter Davey and Andrea Patton for advice on conducting an Interrupted Time Series analysis. \""} +{"text": "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There was an error in the Funding statement. The correct Funding statement is: The study was funded by Martek Biosciences Inc. (There was an error in Table 6. The correct version of Table 6 is available here:"} +{"text": "In the Netherlands, there seems to be a relatively large interest in traumatic stress and its consequences. A literature search for publications in the field of psychotrauma and posttraumatic stress disorder (PTSD) in Web of Science shows that authors from the Netherlands are among the top seven in the world see . Most reIn this thematic cluster we highlight psychotrauma research in the Netherlands. We start with a review of Dutch psychotraumatology (Vermetten & Olff, The aforementioned articles include an article by Diehle et al. , who preMiranda OlffDepartment of PsychiatryAcademic Medical CenterUniversity of Amsterdam & Arq Psychotrauma Expert GroupDiemen, The NetherlandsEric VermettenResearch Center\u2014Military Mental HealthMinistry of DefenseUtrecht, The NetherlandsDepartment of PsychiatryUniversity Medical Center UtrechtUtrecht, The Netherlands"} +{"text": "There were errors in the Funding section. The correct funding information is as follows: \"This work was supported by the National Institutes of Health grant 5DP1DA026197 and Dr. Dean Y. Li was funded by NHLBI. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There was an error in the funding statement. The correct funding statement is: This work was supported by a grant from CIHR China-Canada Joint Health Research Initiative Grant (CCI 102929) to BBY who is the recipient of a Career Investigator Award (CI5958) from the Heart and Stroke Foundation of Ontario. Support for this study was also provided, in part, from the Holland Musculoskeletal Program, Sunnybrook Health Sciences Centre. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There was an error in the funding statement. The correct funding statement is: \u201cThis analysis was funded by the Florida Agricultural Experiment Station. Publication of this article was funded by the University of Florida Open-Access Publishing Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "Information regarding funding for the fourth author is incorrectly omitted from the Funding statement. The Funding statement should read:This work was supported by the Program for Innovative Research Team of Beijing University of Chinese Medicine (2011-CXTD-09) and the 111 Project (No. B08006). Jian-Ping Liu was partially funded by the grant number 2011ZX09302-006-01-03(5) by the Ministry of Science and Technology of China. Eric Manheimer was partially funded by grant no. R24 AT001293 from the National Center for Complementary and Alternative Medicine (NCCAM) of the US National Institutes of Health. The contents of this article are solely the responsibility of the authors and do not necessarily represent the official views of the NCCAM or the National Institutes of Health. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "An author was omitted from the article.Joel A. Spencer was omitted from the author byline; he should be the sixth author and affiliated with the Center for Systems Biology and Wellman Center for Photomedicine, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, United States of America.The contributions of this author are: contributed reagents/materials/analysis tools."} +{"text": "There was an error in the Funding statement.The correct version of the statement is: Pre-doctoral training support to BKS was provided by National Institutes of Health T32 HD043730 and The Foundation for Physical Therapy. Some supplies were provided by an Howard Hughes Medical Institute HHMI-funded mini-grant to BDD. Publication of this article was funded in part by the University of Florida Open-Access Publishing Fund. The funders had no role in study design, data collection, decision to publish, or preparation of the manuscript."} +{"text": "The correct funding information is: \"This study is supported by: Research Grants for PRESTO and SENTAN from JST, Grant-in-Aid for Scientific Research on Priority Areas Molecular Brain Science and Research on Pathomechanisms on Brain Disorders from MEXT. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "In the Funding Statement, the name of a funding organization is incorrect. The Funding Statement should read: \"This study was partly supported by National Natural Science Foundation of China (Grant No. 81270215) and China Postdoctoral Science Foundation funded project (Grant No. 2013M530664). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "This activity was supported by unrestricted educational grants from Stallergenes Allergen Vaccines Worldwide, Nestl\u00e9 Nutrition Institute, and UCB.Stallergenes - http://www.stallergenes.com/nc/en.htmlhttp://www.nestlenutrition-institute.org/enhttp://www.ucb-group.com/and http://www.todaysallergies.comJune 2008Genoa, ITALYDear Colleagues:Innovation in Continuity. WAO will continue to build on its existing programs to further support and develop new research, education and training initiatives in this specialty and to focus attention on the importance of the allergist as the key coordinator of allergy patient care.One of my first duties as President of the World Allergy Organization (WAO) is the pleasure of presenting the State of World Allergy Report 2008. This exciting new WAO initiative, launched at the World Allergy Congress in Bangkok in December 2007, will provide a biennial review of the explosion of allergies around the globe and will consider how the medical, social and economic issues surrounding allergic diseases should be addressed. This first report provides a snapshot of the prevailing situation in 2008 from the perspective of organizations, such as the WAO and the World Health Organization through the Global Alliance against Chronic Respiratory Diseases, which represent allergists and clinical immunologists -- the clinicians at the forefront of attempts to recognize, assess and address this global crisis. The focus of WAO's mandate for the next two years is The State of World Allergy Report is a call to action for everyone with an interest in allergy to develop a partnership with WAO in a global effort to control and manage allergic diseases on behalf of the world's population.Yours sincerely,Professor G. Walter CanonicaPresident, World Allergy Organization"} +{"text": "There was an error in the Funding Statement. The correct funding is:Natural Sciences and Engineering Council of Canada to WJR and MR (grant numbers not disclosed). DM received financial support from the Serbian Ministry of Culture and Ministry of Science, Project 177023. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Vasogliano\" should be \"Visogliano\"A word was misspelled in the Abstract. \"-218\" should be \"-281\"There was an error in the Supporting Information. In Table S1, in the last column of the row \"BH-1 Mandible\", the value \""} +{"text": "There were errors in the Funding statement. The correct statement is: This work was supported in parts by the National Center for Soybean Biotechnology and the Missouri Soybean Merchandising Council to HTN and by a grant (No. AP24-1-0076) from the RIKEN Strategic Research Program for R & D to L-SPT. DTL was supported by RIKEN Foreign Postdoctoral Fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The first and second authors, Xiuyu Chen and Minjie Lu, should be indicated as having contributed equally to the work.In addition multiple funding organizations and grants were incorrectly omitted from the Funding Statement. The Funding Statement should read: \"This study was supported in part by the Research Grant of National Natural Science Foundation of China (81000604 and 81370036 )\uff0cPUMC Youth Fund and the Fundamental Research Funds for the Central Universities (2009-xhj04# and 3332013105), and Grant for Talent Research Star of Fuwai Hospital (2012\u2010FWXX01). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There was an error in the Funding section. The corrected Funding section should appear as follows:This study was supported by grants from National Institute of Health to SJC (www.nih.gov). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There was an error in the Funding statement. The correct version of this Funding statement is available below.This study was sponsored by grants of Zhejiang Provincial Top Key Discipline in Surgery, the Natural Science Foundation of Zhejiang Province (Grant No. Z2100853) and the National Natural Science Foundation of China (Grant No. 81070372). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There were multiple errors and an omission in the Funding Statement. The Funding Statement should read: \"The infrastructure for the NESDA study (www.nesda.nl) is funded through the Geestkracht program of the Netherlands Organization for Health Research and Development and is supported by participating universities and mental health care organizations , Netherlands Institute for Health Services Research (NIVEL) and Netherlands Institute of Mental Health and Addiction (Trimbos Institute). This work was supported by the Top Institute Pharma, project number T5-203. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "In the Funding Statement, an organization was incorrectly as having provided funding for the article. The Funding Statement should read: \"This work was supported by a Building Interdisciplinary Research Careers in Women's Health (BIRCWH) grant and Academic Project Support (APS) funding. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There was an error in the Funding statement. The correct version of the statement is available below:This work was supported by the Society for Research on Cardiovascular Diseases, the Ministry of Culture, Health and Higher Education, and the National Fund for Research of Luxembourg. Profs. Ng and Squire and Dr. McCann received support from the NIHR Leicester Cardiovascular Biomedical Research Unit. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There is an error in the Competing Interests statement. The Competing Interests statement should read: \"AMD is currently a Professor of Pathology, Urology and Oncology at the Johns Hopkins University School of Medicine and was also employed as such during the conduct of this study. During part of the writing of this manuscript AMD was employed at Predictive Biosciences, Inc. , as well as part-time adjunct Professor of Pathology, Oncology, and Urology at the Johns Hopkins University School of Medicine . No funding or other support was provided by Predictive Biosciences, Inc., for any of the work in this manuscript. The terms of the relationship between AMD and Predictive Biosciences, Inc., were managed by the Johns Hopkins University in accordance with its conflict-of-interest policies. This does not alter the authors\u2019 adherence to all the PLOS ONE policies on sharing data and materials.\""} +{"text": "There was an error in the funding statement. The correct funding is:http://www.rcn.no) , the Cancer Foundation of Norway (http://www.kreftforeningen.no) (to KKA), and the NSF-Biochem 0919027 to EIS). The Raman instrument and laser equipment in Oslo was funded by a grant from the Research Council of Norway. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. This work was supported by The PEOPLE Marie Curie Actions Intra\u2013European Fellowship within the 7th European Community Framework Programme (PIEF\u2013GA\u20132009\u2013235237 to GZ), the Research Council of Norway ("} +{"text": "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The Funding statement is incorrect. The publisher apologizes for the error. The correct Funding statement is: This research was financially supported by the National Key Technologies R&D Program (2015BAD07B01), the National Natural Science Foundation of China (31270656,"} +{"text": "There is an error in the funding statement. Please refer to the complete funding statement here:This research was supported by funds from the Cave Conservancy Foundation , Society of Systematic Biologists (Graduate Student Research Award), and the American Arachnological Society (Vincent Roth Fund for Systematics Research) awarded to S Derkarabetian. This research was also supported by an NSF grant awarded to M Hedin (DEB 1354558). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This project was funded in part with Federal funds from the National Institutes of Allergy and Infectious Disease, National Institutes of Health, Department of Health and Human Services, under Contract No. 272200800007C, CTSA award No. UL1TR000445 from the National Center for Advancing Translational Sciences, the Childhood Infections Research Program grant T32-AI095202-01, the Immunobiology of Blood and Vascular Systems training grant 5 T32 HL69765-12, and NIH grant GM064779. The contents of this manuscript are solely the responsibility of the authors and do not necessarily represent official views of the National Center for Advancing Translational Sciences or the National Institutes of Health. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "ZYGX2014Z002. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The Funding statement appears incorrectly in the published article. The correct Funding statement is: This work is partially supported by the National Natural Science Foundation of China ("} +{"text": "The following information is missing from the Funding section: We acknowledge support by the Open Access Publication Fund of the University of Muenster.http://www.nf.mpg.de/). We also acknowledge support by the Open Access Publication Fund of the University of Muenster. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The complete, correct Funding statement is: The Max Planck Institute financed the fMRI study ("} +{"text": "During typesetting, errors were introduced into the Funding section. The publisher apologizes for the error. The correct funding information is as follows: This work was supported by the Canadian Institutes of Health Research (RSN-109427) and the Saskatchewan Health Research Foundation to QL. ZL is a recipient of the University of Saskatchewan Vaccinology and Immunotherapeutics Ph.D. Student scholarship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There is an error in the funding statement. Please refer to the correct funding statement below.This work was made possible by support from the President's Emergency Plan for AIDS Relief (PEPFAR) through Cooperative Agreement 5U2GPS001930-05 from the Department of Health and Human Services (DHHS)/Centers for Disease Control and Prevention (CDC), Global AIDS Program. The findings and conclusions included in its content are solely the responsibility of the author (s) and do not necessarily represent the official position of the Centers for Disease Control and Prevention/Agency for Toxic Substances and Disease Registry. Publication of this article was funded in part by the Open Access Promotion Fund of the Johns Hopkins University Libraries. This work was supported through technical assistance from the Johns Hopkins Center for AIDS Research (1P30AI094189). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The Global Center of Excellence (GCOE) for atomically controlled fabrication technology was established in 2008 by the Japanese Ministry of Education, Culture, Sports, Science and Technology (MEXT), as a succession program of the 21st Century COE program for atomistic fabrication technology promoted from 2003 to 2007. The GCOE program is implemented by three departments, namely the Departments of Precision Science & Technology, Applied Physics, and Advanced Science and Biotechnology, and by the Research Center for Ultra-precision Science and Technology, all of which belong to the Graduate School of Engineering of Osaka University.The Fifth International Symposium on Atomically Controlled Fabrication Technology (ACFT-5) was organized by the GCOE program and the technical committee on ultraprecision machining of the Japan Society for Precision Engineering (JSPE), in cooperation with JSPE, the Japan Society of Applied Physics (JSAP), and the Physical Society of Japan (JPS).The aim of our GCOE project is to achieve the atomic level controllability in wide-area processing and environmental harmony, which are essential for next-generation manufacturing technologies with high functions. For this purpose, by collaborating with other organizations from different fields, we focus not only on the creation of new fabrication processes beyond the current limitations but also on the systematization of the fabrication processes as science.ACFT-5 highlights the recent achievements in the program. The topics covered here are \u2018Impact of Quantum Beam Techniques with Advanced Optics\u2019, \u2018Surface Characterization and Bottom-up Technologies\u2019, \u2018Multi-Scale Surface Control Techniques\u2019, \u2018Advanced Processing and Materials Science for Device Applications\u2019, and \u2018Frontier in Carbon/Organic Nano-Scale Systems\u2019. Finally, we would like to express our gratitude to all the contributors and committee members for their great effort in making the symposium successful and also to MEXT for its continuous support."} +{"text": "Prior to 1993, Nepal had a vertical health logistics system. Logistics was not a government priority. No logistics curricula had been developed, no staff had been trained, and no logistics information systems existed at any level. After the establishment of the Logistics Management Division in 1993, the lack of trained manpower in logistics was realized. With support of USAID funded projects (implemented by John Snow Incorporated through Family Planning Logistics Management (FPLM), Nepal Family Health Program, and DELIVER), logistics training was institutionalized within the National Health Training Centre of Ministry of Health and Population.With support from USAID, the National Health Training Centre and Logistics Management Division have worked to institutionalize logistics training. Trainers were trained and Regional Health Training centres have been conducting logistics training. Logistics training is included in National Health Training Centre\u2019s annual work plan and approved by the National Planning commission. Logistics practices have been incorporated in the pre-service and in-service curricula and health logistics training has also been incorporated in the training management guideline of the National Health Training Centre.Logistics practices have been incorporated in the pre-service and in-service curricula. Technical assistance is being provided to establish or maintain training within a national training system and efforts are being made to build the capacity of government healthcare providers to manage training events. Ten standardized Health Logistics Training packages have been institutionalized into the National Health Training Centre system to train human resource needed for the logistics management of the country and computer based self-paced training (CD-ROM), has been developed.The Ministry of Health has recognized the importance of the not only logistics management but also the need for quality training. The Ministry of Health has initiated and is continuing provision of logistics training from its own financial resources ensuring sustainability of the program to some extent. From 1993 to 2013, a total of 27,734 government personnel have been trained in the health logistics trainings. Through the training important logistics interventions like Pull System of Health Commodities and web-based LMIS were successfully implemented in all 75 districts of the country.Frequent turnover of trained storekeepers and a lack of effective supervision after training remain concerns. The misconception among health workers that training will solve all performance problems hinders their ability to analyse gaps and subsequently address them; and overall governance and accountability of the Government are continued issues."} +{"text": "The following information is missing from the Funding section: National Clinical Key Subject Construction Project Fund of China funded this work. This funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "A second affiliation for the second author is not indicated. Yau-Huei Lai is also affiliated with Mackay Junior College of Medicine, Nursing, and Management, New Taipei City, Taiwan.The following information is missing from the Funding section: This work was partially funded by grants from National Science Council , Mackay Memorial Hospital and Taiwan Foundation for geriatric emergency and critical care."} +{"text": "The following information is missing from the Funding section: This study was supported by Office of Naval Research Multidisciplinary University Research Initiative Grant N000141310672. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There were additional funders for this article that were not included previously. Please refer to the correct funding statement below:This study was supported by a grant of the German medical acupuncture association (D\u00c4gfA), China scholarship council and University Funds from the University Hospitals of Wuerzburg. This publication was funded by the German Research Foundation (DFG) and the University of Wuerzburg in the funding program Open Access Publishing. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Fundamental and clinical experts from Japan, Korea and China, warmly gathered on the beautiful ice and snow city\u2014Changchun, the capital of Jilin Province, China\u2014during 23-26 January 2015, to present their research findings and participated in discussion relating to progress in biomaterials, stem cells and bone tissue engineering. The International Symposium on Recent Trend of Biomaterials and Stem Cells for Bone Tissue Engineering (BTE 2015) was hosted by the Key Laboratory of Polymer Ecomaterials, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, and co-organized by the Department of Orthopaedics, China-Japan Union Hospital, Jilin University. It provided a new platform of academic and technological communication for fundamental researchers and orthopedic surgeons to express their diverse ideas and inspiring new cooperation.The field of tissue engineering has emerged as an important approach to the clinical therapy of damaged bone due to trauma, tumor resections or congenital anomalies . Besidesin\u00a0vivo microenvironment is a promising strategy for tissue engineering and stem cell research [The participants focused on discussing the recent development of synthetic or natural bioscaffolds that can provide 3D architecture , 9, apprresearch . A serieresearch or peptiresearch have beeresearch , 19. TheWe made a concise report on this symposium, and some comments of the key speakers in the symposium were presented. We hope that their comments can arouse further interests of readers in discussing future strategies of bone tissue engineering and regenerative medicine. Moreover, we believed that the frequent appointment of researchers and orthopedic surgeons may inspire more and more ideas in the development of future clinical products of tissue engineering for orthopedic application.Chief Scientist and Director of the Nano Medical Engineering Laboratory at the RIKEN (from 2004). He received his Bachelor\u2019s degree in polymer chemistry at Kyoto University and was awarded a doctorate in engineering from the same university in 1987. Since then he has held some posts as assistant and associate professor at Kyoto University, research fellow at the University of California, Irvine (1992-93), professor of the University of Tokushima, and Project Leader at the Kanagawa Academy of Science and Technology. Currently, he is also a visiting professor of some universities including Tokyo Institute of Technology, Tokyo Metropolitan University, Hokkaido University and Waseda University. He received the Award of Japanese Society of Biomaterials (2009) and Fellow, Biomaterial Science and Engineering (2012). His research focuses on biomaterial science, regenerative medical engineering, combinatorial bioengineering for the creation of functional polymers and soft nanotechnology.Comment: The surface properties are of prime importance in establishing the response to tissue from biomaterials. I have investigated the immobilization of growth factor proteins to provide a set of very powerful signals for cells. Immobilized growth factors stimulated cultured cells for a longer time and enhanced cell growth more than did soluble growth factors and also induced cellular differentiation. The effects of immobilized growth factors were visualized by their micropatterning on substrates. Recently, we extended the immobilization technology from chemical to biological one, which is protein engineering for synthesis of binding growth factors. We synthesized some types of growth factors carrying collagen binding sequence extracted from fibronectin. One of them was collagen binding-BMP, which was synthesized by silk worm, and directly injected or utilized with scaffold or visible-light-curable gelatin gel. We demonstrated that both of the direct injection and the bound implants significantly enhanced bone formation in animals. Another type of binding growth factors was also designed by combination with binding motif of underwater adhesive proteins composed of non-canonical amino acids such as phosphorylated serine from salivary statherin and 3,4-dihydroxyphenylalanine from mussel protein for binding to inorganic materials such as metals and ceramics. The technology uses not only usual protein engineering but also bioorthogonal approach. As a result osteogenetic differentiation was enhanced on the bound apatite. These new types of binding growth factor could contribute current treatment methods of various debilitating injuries and diseases and deliver on the therapeutic promise of bone tissue engineering.Professor in the Department of Polymer Science and Engineering at the School of Applied Chemical Engineering, Graduate School, Kyungpook National University. He is currently a director of Center for Intelligent Hybrid Nanomaterials and Processes supported by Brain Korea 21+ National Program. He obtained his Ph.D. degree in Polymer Chemistry at Kyoto University in 1987, and joined in the Department of Polymer Science and Engineering in 1988 as a professor. He has published more than 190 articles and his research is focused on surface modification, nanofiber scaffolds, functionalization of nanoparticles and liquid crystal sensors.Comment: During last one decade several advancements have been made in fabrication of scaffolds for tissue engineering using various techniques of fabrication and materials of different origin. The scaffolds for tissue engineering are ranging from nanofibers to nanoparticles and have been developed by tuning their morphology and architecture. Several breakthroughs have been reported and successful attempts have been made to overcome the drawbacks of the scaffolds fabricated by conventional and modern methods of scaffolds formation yet significant amount of research is required to control the vascularity of the scaffolds, which is a major challenge in the field of fabrication of scaffolds for bone tissue engineering. Although few attempts have been made by harvesting cells from donor/patient to vascularize the scaffolds before their implantation in the patient but area still need intensive research to develop scaffolds with proper vascularization. The worldwide research has achieved success in tissue engineering but matrix induced autologus condrocyte implantation has received little success in repair of cartilage. The bioactivity of scaffolds totally depends on surface topology, microstructure, chemistry and mechanical properties of scaffolds. Thus, controlling the cells behavior and remodeling is a critical step in development of next generation scaffolds for tissue engineering. The functionalized scaffolds loaded with certain amount of growth factor and therapeutics to stimulate cells proliferation and treatment of infection after surgical operations also strongly needed to achieve success in the area of tissue engineering and regeneration. The increasing interest to incorporate a drug delivery function in the scaffolds is emerged as a new field of scaffolds engineering. The drug loaded scaffolds are able to provide sufficient amount of drug at a specific place in comparison to systematic drug delivery systems. The scaffolds loaded with growth factors such as cytokines, hormones, morphogens and proteins need to be developed to enhance cells adhesion, proliferation and differentiation in tissue engineering. The mechanical behavior of scaffolds has important implication on tissue regeneration; stiff scaffolds mimic precalcified bone and elastic scaffolds mimic the elasticity of muscles, which cause MSC differentiation down to a myogenic pathway. Thus, scaffolds with optimized mechanical strength need to be fabricated for enhanced bone tissue engineering. The critical review of the literature reported during last one decade has clearly indicated that there is lot of scope to improve the properties and functionality of the scaffolds for bone tissue engineering using various biomaterials and their fabrication techniques.Principal Investigator and Unit Director of Tissue Regeneration Materials Unit, International Center for Materials Nanoarchitectonics, National Institute for Materials Science, Japan. He obtained his Ph.D. from Kyoto University in 1997 majoring in biomaterials and did postdoctoral research until 2000. He became researcher in 2000 and senior researcher in 2003 at Tissue Engineering Research Center, National Institute for Advanced Industrial Science and Technology, Japan. He moved to Biomaterials Center, National Institute for Materials Science as senior researcher in 2004 and was group leader from January, 2007 to March, 2011. He is concurrently Professor of Joint Doctoral Program in Materials Science and Engineering, Graduate School of Pure and Applied Science, University of Tsukuba, Japan. He is Guest Professor of Shanghai Institute of Ceramics, Chinese Academy of Sciences; Southeast University and Sichuan University, China. He is Associate Editor of Journal of Materials Chemistry B; Editor of Science China Chemistry and Editorial Board of Journal of Bioactive and Compatible Polymers and Tissue Engineering .Comment: Tissue engineering has attracted tremendous endeavors from various fields of material science, biomedical science, chemical engineering, stem cell biology, clinics, industry and etc. The disciplinary research to regenerate functional tissues and organs has reached enormous progress and achievements spanning form fundamental research to clinical applications and industrialization. As one of the tissue engineering, triad, scaffolds not only accommodate implanted cells but also control cell functions to guide functional tissue engineering. Design and creation of highly functional scaffolds have been challenged to mimic the in\u00a0vivo dynamically remodeling cellular microenvironments. Scaffolds with controlled chemical compositions, nano- and micro-structures, mechanical property and some special functions have been developed form synthetic and naturally derived biomaterials and decellularized matrices. The scaffolds can provide biological and physiochemical cues to recruit stem cells, induce stem cells differentiation and promote regeneration of various tissues and organs. Development of new and highly functional scaffolds will further facilitate efficient regeneration of functional tissues and organs for clinical applications and industrialization.Professor of Chemical and Biological Engineering and a Chairman of Interdisciplinary Program of Bioengineering in Seoul National University, South Korea. He is an active researcher with 275 peer-reviewed publications, 14 book chapters and 17 patents. He has received William B. Walsh Award and Shinyang Award. He had served or now serves as an Associate Editor or an editorial board member of 10 journals including the Tissue Engineering, Journal of Tissue Engineering and Regenerative Medicine, Biotechnology and Bioprocess Engineering, Journal of Industrial and Engineering Chemistry and Tissue Engineering and Regenerative Medicine. He is the chair of Scientific Committee of Korean Tissue Engineering and Regenerative Medicine Society and an organizer of 15th International Biotechnology Symposium.Comment: Currently, a number of bone graft substitutes have been developed and are being utilized clinically, but there are rooms for improvement. Bone graft substitutes can promote bone regeneration by the interaction of three components: (i) bone-forming cells, (ii) a suitable biomaterial scaffold and (iii) biological stimulants. In the International Symposium on Recent Trend of Biomaterials and Stem Cells for Bone Tissue Engineering (BTE 2015), which was held in Changchun in China in January, 2015, new technologies on these three components for bone regeneration were presented. For large bone-defect, stem cells or progenitor cells need to be implanted since recruitment of bone-forming cells to the large bone-defect may be limited. For effective bone regeneration, implantation of osteogenically induced stem cells or progenitor cells results in better bone regeneration rather than implantation of undifferentiated cells. Thus, decent technologies for ex vivo osteogenic induction of stem cells would be necessary. Meanwhile, biomaterials can be designed in nano scale to present biological stimulants for effective bone regeneration. BMPs are currently used for bone regeneration in patients, but often cause adverse effects due to the high dosage. Drug delivery carrier determines the bone regeneration efficacy of BMPs. Efforts should be made to develop an appropriate delivery carrier which can deliver BMPs without the adverse effects.Professor of polymer chemistry in Changchun Institute of Applied Chemistry (CIAC), Chinese Academy of Sciences (CAS) from June 1999. He graduated from the Chemistry Department of Jilin University. He got his Master degree from CIAC in 1988. He got his Ph.D. from Waseda University Japan on March 1997. He had postdoc experience in the University of Pennsylvania from 1997 to 1999. Now he is working in Key Laboratory of Polymer Ecomaterials of CAS in CIAC. He is one of the board members of Journal of Controlled Release, Biomacromolecules, Advanced Healthcare Materials, Acta Biomaterialia and Macromolecular Bioscience. He has published 490 papers.Comment: In the past 20 years, the regenerative medicine materials, especially the tissue engineering repair materials have made important progress. Tissue engineering is an interdisciplinary field, including materials chemistry and physics, biological engineering, biochemistry, biomechanics, cell biology, biotechnology, clinical medicine, etc. It is one of the important means to repair and rebuild the body\u2019s tissues and organs and has been becoming more and more important. The novel therapy method will change the quality and the way of people\u2019s life, and extend the life of human beings in the near future. Biodegradable polymer material (absorbed in\u00a0vivo) is one of the very important elements in the process of tissue engineering therapy. As a result, the preparation of porous bioscaffolds with adjustable absorption time, good strength, pore structure and distribution under well control, and good biocompatibility, are the objective requirements for the future development of tissue engineering therapy. We hope that through the joint efforts of scientists, more and more products of tissue engineering devices will be widely used in clinic in the near future.The \u2018Qiu-shi\u2019 Distinguished Professor and dean for School of Basic Medical Science, Vice Dean College of Medicine in Zhejiang University, China. He received his Ph.D. degree in orthopedics from National University of Singapore in 2003. Since returned to China in 2005, Prof. Ouyang focuses on the research of tissue engineering and regenerative therapy of musculoskeletal system. His research has been granted support by National Natural Science Foundation of China (NSFC) for Distinguished Young Scholars, The State Key Program of National Natural Science Foundation of China, The National Key Basic Research Program, The National High Technology Research and Development Program of China (863 Program), etc. Currently, he has filed 10 national patents applications (six approved) and published more than 60 original research papers in the international peer review journals. More than 40 papers corresponded by Prof. Ouyang were published in the leading journals of regenerative medicine fields, such as Stem Cells, Advanced Functional Materials/Biomaterials, Annals of the Rheumatic Disease, Tissue Eng, Cell Transplantation. Moreover, he was authorized by State Food and Drug Administration (SFDA) and Ministry Of Health (MOH) to formulate the guidance for assessment of implants for cartilage repair and therapeutic transplantation of engineered tissues, respectively. Prof. Ouyang has established a standard approach for tissue engineered cartilage (TEC) transplantation and is the pioneer of clinical translation in China orthopedic regenerative medicine. Because of his outstanding achievements, Prof. Ouyang has gained great national and international reputation, he is currently the chair of China Orthopedic Regenerative Medicine and the vice chair of China Tissue Engineering and Regenerative Medicine Society. He is also the council member of several international academic societies such as Tissue Engineering and Regenerative Medicine International Society-Asia Pacific Meeting (TERMIS-AP).Comment: Decades of experience indicated that manufacture of mature tissues with tissue engineering approaches for transplantation is difficult to be achieved. The most feasible and effective way is to apply one or all of the appropriate cell source, biomaterials/scaffold, growth factors according to particular tissues and diseases at the early or middle stage of diseases, which can prevent the tissue/organ from failure and avoid organ transplantation.Professor of Department of Chemistry, Tongji University, China. He obtained Ph.D. at the Shanghai Institute of Ceramics (CAS) in 2005, after which he worked as a postdoc in Hong Kong University of Science and Technology, University of Tokyo, and RIKEN. In 2011, he was appointed as a professor at Tongji University. His research interests focus on the mild polymerized preparation of tough nanocomposite gel and the potential applications in tissue engineering and bio-imaging. His current group has published more than 20 Science Citation Index papers, including Advance Materials and Chemical Science.Comment: Hydrogels with synthetic or natural polymer are useful in cartilage tissue engineering due to their water-swollen networks and suitable microenvironment for tissue formation. Up to now, engineering cartilage tissue still remains a significant challenge due to the low mechanical strength and unsuitable component design. The in-situ hydrogelations via the mild enzyme-mediated radical polymerization can provide a tough artificial cartilage by the direct injection in the defect position.Professor in Shanghai Institute of Ceramics, Chinese Academy of Sciences . He completed his Ph.D. in 2006, and then he worked in the University of Sydney, Dresden University of Technology, Germany and Queensland University of Technology where he was awarded Vice-Chancellor Research Fellow, APDI Fellow and Alexander von Humboldt Fellow. In 2012, Dr. Wu has been recruited to work in SIC, CAS, as One-Hundred Talent Program of Chinese Academy of Sciences. Then he was awarded Recruitment Program of Global Young Experts of China and Shanghai Pujiang Talent Program. Prof. Wu\u2019s research focuses on bioactive inorganic materials for bone tissue engineering and drug delivery application. Up to now, Prof. Wu has published more than 110 SCI peer-review journal papers, including Adv Funct Mater, Biomaterials (14 Papers), Adv Mater Interface, Small, J Control Release, J Mater Chem (15 Papers), Acta Biomater (22 Papers), ACS Appl Mater& Interface, Bone, Tissue Eng. etc. The papers have been cited more than 2300 times, H Index 31. Prof. Wu has been awarded 11 patents, in which three of them have been transferred to company.Comment: Regeneration of large-size bone defects represents a significant challenge clinically, which requires the used scaffolds with the stimulation of osteogenesis and angiogenesis for stem cells. It is known that microenvironments of bioscaffolds play an important role to stimulate tissue regeneration. How to design and prepare bioscaffolds with favorable microenvironments for tissue regeneration is one of interesting topics in the fields of biomaterials and tissue engineering. To establish a beneficial microenvironment of biomaterials, it is of great importance to combine both nutrient elements and biomimetic structure of scaffolds to stimulate osteogenesis and angiogenesis of stem cells. Besides the microenvironment of bioscaffolds, macrophage plays an important role to induce immunomodulatory microenvironment for further influencing osteogenesis. Therefore, it is suggested that scaffolds and osteoimmunomudulation-induced microenvironments should be considered for bone tissue engineering.et\u00a0al. Currently, he has published over 60 original research papers.Dr. Zou is the director of the Orthopedic Research Institute and consultant surgeon of Spine Division at the First Affiliated Hospital of Sun Yat-sen University (SYSU). He graduated from Jiangxi Medical School, receiving the doctoral degree in Medical School of Aarhus University, Denmark. In 2008, Xuenong has been recruited as professor of the First Affiliated Hospital as One-Hundred Talent Program of SYSU. He has been focused on the interaction between biomaterials and host, the effect of ECM-biomaterials on bone formation, epigenetic mechanisms of osteogenetic progress induced by different biomaterials. His research has been supported by The State Key Program of National Natural Science Foundation of China, The National Key Basic Research Program (\u2018973\u2019 Program) and The Key Program of NSFC-Guangdong Joint Fund, Comment: There exist three sequential biological events after implantation of biomaterials\u2014stress and homeostasis, immuno-inflammatory responses and bone formation or implantation failure. The clinical result of the implantation depends on the first two events, happening in the interaction between biomaterials and human body. Therefore, it\u2019s very important to elucidate the roles and mechanisms of early biological events caused by implantation of biomaterials, as a guide for synthesizing and designing biomaterials.Professor of Department of Orthopedics in China-Japan Union Hospital, Jilin University, China. He graduated from the Norman Bethune Medical University in 1991. He got his Doctor degree from Jilin University in 2003. He had his visiting scholar experience in the University of New South Wales in Australia from 2008 to 2009. He has been engaged in orthopedic clinic for more than 20 years. His main research work is on spine surgery and bone tissue engineering and has published more than 60 papers. Now he is the member of the expert committee of Chinese national medical examination center, the member of the Chinese Medical Association Branch of minimally invasive Department of orthopedics and one of the China core members of Medtronic MMSG Association.Comment:At present, there are about 3 million patients with defect bone or injured bone needed to be treated each year in China. More and more biological materials are demanded in orthopedic clinical work. To meet substantial clinical demand, with the high speed promotion of biomedical materials science and engineering, a high-tech biomedical materials and its products industry has already formed. But the present products of biomaterials can\u2019t really biomimic the autologous grafts and the surgeons usually wondered that which was the best choice for a case when they face to a wide variety of products. Some of them will worry about the dissatisfied or even failed treatment and then refuse to accept the new products. One possible reason to affect the bone healing quality of a product might be the selection of operation indication or operation method but not the product itself. Thus, it is important for both researchers and surgeons that their frequent communication and cooperation will be conducive to the development of new biomaterials and the rapid development of orthopedic clinical technology."} +{"text": "The National Plan of Prevention PNP 2014-2018) [4-2018 1 acknowleContinuous training for health professionals started as a national program in Italy in 2002; some years later (2007) administrative control had been transferred from the Ministry of Health to the National Agency for Health Services, Agenas. At distance training and online updating are now becoming very common, close to traditional kinds of events for training, always in respect of the Evidence Based Medicine (EBM).We can identify, in the continuous sanitarian training system, different roles and various kinds of providers. Relevant organisations include the National Committee for Training, in the Ministry of Health, the Warranty Committee, and the National Observatory about Quality of Training.Regarding research in the field of health, we can consider it as the architrave of the National Health Service; the Report on Health Status in Italy 2012-13 defines The National Committee for Sanitarian Research defines the program and initiatives, monitors and evaluates the results. Research institutes involved in Italy must accept different challenges, according with the Singapore Declaration , which dThe European Union invested 80M Euro in research with Horizon 2020, in the aim of destroying barriers and realising, all over the world, a common environment concerning knowledge, research and innovation.A recent piece of research related to the food habits in the age of complementary feeding is Nutrintake 6/36 ["} +{"text": "The correct funding information is as follows: This work was supported by the Sall Family Foundation via the Pan American Health and Education Foundation (renamed PAHO Foundation) and EUROHEALTH. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. URL: The publisher apologizes for these errors."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This work was supported by National Institute of Health Research Grants R01DE10742 and R01DE14183, and The Mary Kay Foundation Research Grant number 005\u201313 (to DKA). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The Funding Statement is missing funding information from the NIH. Please see the full Funding Statement here.This study was funded by the Burroughs Wellcome Fund Award for Investigators in Pathogenesis (EEH) and the NIH T32-AI007422 (HGB). This work is based upon research conducted at the Northeastern Collaborative Access Team beamlines, which are funded by the National Institute of General Medical Sciences from the National Institutes of Health (P41 GM103403). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The following information is missing from the Funding section: This research was supported in part by Judy Polshek Goodman Alzheimer's Disease Research and Prevention Fund, and a pilot grant from Philips Imaging.The complete, correct funding information is as follows: This research was supported in part by the Spitz Brain Health Innovation Fund, the G.R. Lincoln Family Foundation, Judy Polshek Goodman Alzheimer's Disease Research and Prevention Fund, and a pilot grant from Philips Imaging. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The publisher apologizes for the error."} +{"text": "The following information is missing from the Funding section: This work was supported in collaboration with the National Science Foundation, Partnerships for International Research and Education (OISE-0530174).Vibrio spp., and E. coli.There is an error in the first sentence of the third paragraph in the Introduction. The correct sentence is: In a study of bacteriological assessment of ballast waters of six ships docked at Singapore harbors, Joachimsthal et al. (2004) used fluorescence in situ hybridization (FISH) coupled with flow cytometry to enumerate total bacteria, and fluorescent tags to differentiate between Enterobacteria, There is an error in the caption for Please see the complete, correct"} +{"text": "There is a missing grant number in the funding statement for this article. Please refer to the correct funding statement below:Research at QHFSS was funded by an internal funds provided by Queensland Health and the US National Institutes of Health/NIGMS U01GM097661. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The identification number of the project was incorrect. The article was supported by the project DEC-2011/03/N/NZ5/00248 funded by National Science Centre.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "There is an error in the affiliation for authors Mosoka Fallah, Josephine Monger, and Kodjo Tehoungue. The correct affiliation is: the World Health Organization."} +{"text": "There is an error in the grant number in the Financial Disclosure (FD) statement for this article. Please refer to the correct FD statement below:This study was supported by grants 81071095, 81120108011, and 81361128010 (to XX) from the National Natural Science Foundation of China, Suzhou science and technology development program (SYS201232), the Priority Academic Program Development of Jiangsu Higher Education Institutions of China, Jiangsu Province\u2019s Key Provincial Talents Program (to XX), and the National Basic Research Program of China . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "RRM was omitted from a grant in the Funding section. The correct Funding information is: This work was supported by the National Institute for Health Research , funds for consumables and WellChild (RRM and AD), funds were given to purchase the encapsulator used in the work. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There is an error in the Funding section. The correct funding information is as follows: The work was supported in part by grants from the Australian Research Council, Cancer Council Queensland to J.N. and the Internal Grant Agency (IGA) of the Ministry of Health of the Czech Republic (IGA NT/14078-3), and the European Regional Development Fund to JN. EAP was supported by the Douglas Francis Green PhD Scholarship provided by the Queensland Asbestos-Related Disease Society. LFD was supported by the Griffith University Research Fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This research was supported by the Hi-tech Research and Development Program of China (Grant No. 2012AA10A408 and 2012AA092203), the National Natural Sciences Foundation of China (Grant No. 31072245 and 31372567), the special foundation under the Construction Programme for \u2018Taishan Scholarship\u2019 of Shandong Province of China, and Special Scientific Research Funds for Central Non-profit Institute, Yellow Sea Fisheries Research Institute (20603022013010). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: Research was supported by the Czech Science Foundation (206\u201308\u20130640), the Ministry of Education, Youth and Sport of the Czech Republic (SVV260 087/2014), and the National Science Foundation (DEB0746560). Mouse pellets and genotyping of allopatric mice were reimbursed from the Czech Science Foundation (P506\u201311\u20131792). The funders had no role in study design, data collection, data analysis, decision to publish, or preparation of the manuscript."} +{"text": "Information is missing from the Funding section. The correct funding information is as follows: This work was supported by grants from the National Institutes of Health to CIG , JGF (PR-LSAMP and RISE Award 2R25GM61151), MHP and MM (R25GM061838). This publication was made possible by Grant P20 RR 016174 from the National Center for Research Resources, a component of the National Institutes of Health. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "To the Editor: After not finding any additional cases of Middle East respiratory syndrome (MERS) for several weeks in South Korea, in July 2015, the South Korean government and the World Health Organization (WHO) discussed the appropriate time to declare the end of the outbreak in July 2015 . All possible contact with diagnosed case-patients in the late phase of the MERS outbreak in South Korea were traced (Epidemiologic data, probabilistic model, and supplementary discussion."} +{"text": "In Europe, colorectal cancer (CRC) is the second most common cancer, and the second most common cause of death from cancer . The vasGuidelines for staging of CRC have been developed by the European Society of Medical Oncology (ESMO) , which r"} +{"text": "William B. Karesh is not included in the author byline. He should be listed as the tenth author and affiliated with Wildlife Health and Health Policy Program, Wildlife Conservation Society, Bronx, New York, United States of America. The contributions of this author are as follows: Collected and provided fecal samples analyzed in the manuscript."} +{"text": "After publication of this work , it has \u201cThe views and opinions expressed therein are those of the authors and do not necessarily reflect those of the HTA, NIHR, NHS or the Department of Health.\u201dThe full acknowledgements section can be found below:This project was funded by the National Institute for Health Research Health Technology Assessment (NIHR HTA) Programme (project number 10/104/06). Thank you to the trial coordinating staff at the LSHTM CTU. The views and opinions expressed therein are those of the authors and do not necessarily reflect those of the HTA, NIHR, NHS or the Department of Health."} +{"text": "Conflict of Interest StatementES and AS are the creators of the software package Dr. Neuronowski\u00ae, realized as part of a project at the Nencki Institute with funding from the National Centre for Research and Development in Poland. The rights to the software lie with the Nencki Institute that has an agreement with Harpo Ltd., the company commercializing this software. ES and AS are not the owners of this technology nor do they have a direct financial arrangement with Harpo Ltd.The authors state that the correction does not affect the scientific validity of the results.ES: experimental design, data acqusition and analysis, manuscript writing. AD: data acqusition and analysis, manuscript writing. AS: data acqusition and analysis, manuscript writing. TW: guidance on MRI data analysis. AS: subject recruitment. ID: subject recruitment. AO: data acqusition and analysis, manuscript writing.The other authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The second and third authors affiliation was incorrectly stated as both the University of York and the University of Sheffield, which requires amendment . This correction does not in any way affect the scientific validity of the results.GP conceived of, and designed, the study in consultation with PT, LE, and EM. GP conducted the study and analyzed the data with assistance and contributions from PT. GP drafted the manuscript with contributions from PT, LE, and EM. All authors read and approved the final manuscript.This research was supported by the Economic and Social Research Council [grant number ES-J500215-1] awarded to GP whilst at the University of Sheffield.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The following information is missing from the Funding statement: This work was also supported by an NSF award (MCB-1051350) to MCY.The complete, correct Funding statement is as follows: This work was supported by a Scientist Development grant (0830279N) from the American Heart Association and an NSF award (MCB-1051350) to MCY. The publication costs of this article were supported by the Julian Park Fund, College of Arts and Sciences, University at Buffalo. The funders had no role in study design, data collection, data interpretation, or the decision to prepare and publish of the manuscript."} +{"text": "Authors who received the funding were CE and LF. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There are errors in the Funding section. The correct funding information is as follows: The 2015 data collection was supported by the SeaWorld and Busch Gardens Conservation Fund ("} +{"text": "There is an error in the Financial Disclosure (FD) statement for this article. Please refer to the correct FD statement below:http://www.cdc.gov/cancer/npcr/) to the Puerto Rico Central Cancer Registry (PRCCR), by the University of Puerto Rico Comprehensive Cancer Center and by the Department of Health Services Administration, Graduate School of Public Health, Medical Sciences Campus, University of Puerto Rico. The ideas and opinions expressed herein are those of the authors, and endorsement by the funders is not intended nor should it be inferred. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.This work is supported by federal funds from the National Program of Cancer Registries (NPCR Award Number 5U58-DP 003863-02; URL:"} +{"text": "In the abstract, the name of cell line appears as SCH7072. The name of this cell line should read as SCH07072.The Funding statement should be corrected to read as follows: This study was supported, in part, by a grant from the Agenda (No. PJ0102012014) from the Rural Development Administration (RDA) of Korea and the Korea Institute of Ocean Science and Technology project (No. PE99154) of Korea. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Sirs,et al. published in June 2014. We are conscious that, since the study was undertaken in 2010, the public health research landscape in the UK has developed considerably. A number of programmes were set up in 2009 which now have strong portfolios of public health research. All parts of the National Institute for Health Research (NIHR) support research in public health to provide significant opportunities for researchers and evidence for practitioners and decision-makers . Public health research is particularly funded by the NIHR through the Public Health Research programme, focusing on interventions outside of health care, and the School for Public Health Research. The Health Technology Assessment programme evaluates NHS interventions, while Programme Grants for Applied Research supports NHS research delivering findings with early practical application. While the remit varies by programme, covering England or the UK, the application of research findings could extend across Europe. Research for public health is also supported by other Department of Health funding schemes, such as the Policy Research Programme and other funding bodies; for example, a recent option for early-phase intervention research is the Medical Research Council's Public Health Intervention Development scheme. NIHR outputs are captured in a suite of five peer-reviewed, open access journals for anyone to use . We welcome contributions to identify research needs, deliver research to answer key questions and bring new evidence back into practice to strengthen public health research in the UK for the future.We read with interest \u2018Public health research in the UK: a report with a European perspective\u2019 by McCarthy"} +{"text": "A population-based cancer registry has been used for the planning and evaluation of cancer control activities based on administration and the care of individual cancer patients by those in the medical profession. The Japanese Breast Cancer Society (JBCS) registry was started in 1975. In 2004, the registry system was moved to a new system using web registration with the cooperation of the Non-Profit Organization Japan Clinical Research Support Unit and Public Health Research Foundation . Comprehensive individual patient data were recorded according to the Unio Internationalis Contra Cancrum (UICC) TNM classification . Annual We herein report the results of a 5-year prognostic analysis of cases registered in 2005"} +{"text": "In the Funding section, the grant number 61370109 was omitted from the National Science Foundation of China. Please view the complete, correct Funding statement here.This work was supported by the National Science Foundation of China under Grant Nos. 61272339 & 61370109, the Natural Science Foundation of Hubei Province under Grant No. 2012FFB04204, and the Key Project of Anhui Educational Committee, under Grant No. KJ2012A005. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The Academic Editor field does not appear on this article. The Academic Editor is Stuart Humphries, University of Lincoln, United Kingdom. The publisher apologizes for the error."} +{"text": "The affiliation for the fourth author is incorrect. Ruoling Chen is not affiliated with #3 but with #4 Centre for Health and Social Care Improvement (CHSCI), University of Wolverhampton, Wolverhampton, United Kingdom."} +{"text": "A funder was omitted from the Financial Disclosure of the published article. The complete, correct, Financial Disclosure is: This work was partially supported by the Scientific and Technological Research Council of Turkey (TUBITAK) Project No: 113M237 and Bogazici University Scientific Research Fund (BAP) Project No: 9360. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There are errors in the Funding section. The publisher apologizes for the error. The correct funding information is as follows: GC was supported by the European Union European Regional Development Fund through the Centre of Excellence in Genomics to Estonian Biocentre and University of Tartu, by Estonian Personal grants PUT-766 and Estonian Institutional Research grants IUT24-1.The Acknowledgement section is missing from the text. The Acknowledgements are as follows: We thank both of the anonymous reviewers for their constructive criticism. Computational analyses were carried out at the High Performance Computing Center (HPC), University of Tartu and the Core Computing Unit of the Estonian Biocentre."} +{"text": "As a follow up to OpenCon 2014, International Federation of Medical Students\u2019 Associations (IFMSA) students organized a 3 day workshop Open Access, Open Education Resources and Open Data in Kampala from 15-18 December 2014. One of the aims of the workshop was to engage the Open Access movement in Uganda which encompasses the scientific community, librarians, academia, researchers and students. The IFMSA students held the workshop with the support of: Consortium for Uganda University Libraries (CUUL), The Right to Research Coalition, Electronic Information for Libraries (EIFL), Makerere University, International Health Sciences University (IHSU), Pan African Medical Journal (PAMJ) and the Centre for Health Human Rights and Development (CEHURD). All these organizations are based or have offices in Kampala. The event culminated in a meeting with the Science and Technology Committee of Parliament of Uganda in order to receive the support of the Ugandan Members of Parliament and to make a concrete change for Open Access in the country. Librarians, academia and researchers in Uganda have shown a continuous and resolute interest in the Open Access movement. Open Access journal publishing initiatives and support to Open Access repositories can prove it. The Makerere University Library, for example, was the first library in Uganda to set up an institutional repository called the Uganda Scholarly Digital Library (USDL). Launched as a science repository but later changed to cover other disciplines, USDL has a total of 1,600 full text articles, reports, posters, and other scholarly materials [Next Generation of Medical Researchers workshops\u2019. The previous two were pre- General Assembly workshops focusing on Research Integrity. To follow up with those pre-GAs, more workshops have been planned for the regional meeting as well and As Africa was the first region to host the meeting, we decided to focus on: a core part of research data; on educational resources fundamental for students and researchers; and, on research outputs and how to share them. Our main aim is to build capacities to launch and sustain Open Access, and OD (Open Data) activities, projects and policies in the region, especially at national and local level. For this reason we decided to create a network in Uganda with local Universities and personalities based in the capital of Uganda, Kampala. The 10th African Regional Meeting was held in Kampala, Uganda under the theme Human Resources for Health: A foundation for Universal Health Coverage and was organized by the Federation of Uganda Medical Students Association (FUMSA). We received support from: Consortium for Uganda University Libraries (CUUL), The Right to Research Coalition [The IFMSA participated in the OpenCon 2014 from 15-17 November 2014 in Washington DC, and the representatives came back with lot of enthusiasm and a willingness to share their knowledge and new ideas with the medical students. In order to follow up with the OpenCon 2014 a pre Africa Regional Meeting Open Access, Open Education Resources and Open Data was held in Kampala, Uganda from 15-18 December 2014. This pre- ARM workshop was organized within the series of \u2018The students had the chance to meet and be trained by University Librarians from the International Health Sciences University and Makerere University Business School. The librarians are: Florence Mirembe who is also the Chairperson of the Consortium of Uganda University Libraries (CUUL) which promotes access to e-resources to support education and research within universities and other research institutions, and she is also the EIFL (Electronic Information for Libraries) country coordinator; Alison Kinengyere who is also the outgoing president of Association of African Health Librarians-Uganda Chapter and Hassan Segooba who is a librarian at the Makerere University Business School Library. In this way medical students can join their effort to promote Open Access. The participants also had the chance to get also the perspective of a medical journal editor thanks to the participation of Allan Mwesiga from the Editorial Office of the Pan African Medical Journal who shared the experience and achievements of an African Medical Journal as an Open Access journal. Under the guidance of Nsereko Ibrahim from the Centre for Health Human Rights and Development (CEHURD) participants learned more on promoting Open Access and on how to organize initiatives in Uganda, but also in other Universities with A case for CC licensing opportunities in Uganda.Through this event facilitated by Meggie, Agostinho and Mohamed, the students developed critical skills as well as advocacy and practical skills and had the opportunity to share their backgrounds, their project, how they have been involved in OA, OER and OD and analyse examples of successful projects with a view to planning on how to improve them or how to adapt them to their backgrounds. They are now looking forward to spreading awareness about Open Access. The event culminated in a meeting with the Science and Technology Committee of Parliament of Uganda, where the workshop participants presented a statement. There will be a follow up on this with the Ugandan Members of Parliament in order to receive their support and to make a concrete change in the country."} +{"text": "The following funder is missing from the Funding section: Deutsche Herzstiftung. The correct funding information is as follows: This work was funded by the Deutsche Forschungsgemeinschaft (DFG) and the Deutsche Herzstiftung. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The original Supporting Information files were published in error. The files were removed from the HTML and PDF versions of this article on March 11, 2016. The publisher apologizes for the error."} +{"text": "The following information is missing from the Funding section: This work was supported by the European Union Seventh Programme for Research, Technological Development and Demonstration under the grant agreement STREPSYNTH (project No. 613877). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: The authors acknowledge the financial support of the Bankwest Curtin Economics Centre and the Cooperative Research Centre for Living with Autism Spectrum Disorders (Autism CRC), established and supported under the Australian Government's Cooperative Research Centres Program. The authors acknowledge the financial support of Curtin University to Melissa Scott through the Australian Postgraduate Award Scholarship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There is information missing from the Acknowledgments. The Acknowledgments should read: Our sincere thanks go to Wilson Waters, the software engineer from Alintech. The online version of the Q sort would not have been possible without your development and innovation. We are grateful to all the participants, their families and businesses who participated. A special mention goes to EDGE Employment Solutions, EPIC Employment Service, Barkuma\u2019s Personnel Employment in South Australia, Autism Spectrum Australia and AIM Employment of the Autism Association of Western Australia, for their assistance with participant recruitment in this study.The authors acknowledge the financial support of the Bankwest Curtin Economics Centre and the Cooperative Research Centre for Living with Autism Spectrum Disorders (Autism CRC), established and supported under the Australian Government's Cooperative Research Centres Program."} +{"text": "The meteorological data was used to assess the PV, wind and hydropower system potentials on the islands. Furthermore, the reconnaissance study for hydro-potentials were conducted through topographic maps in order to determine the potential sites suitable for development of run-of-river hydropower generation. The stream data was collected for 14 islands in the South China Sea with a total of 51 investigated sites. The data from this study are related to the research article Specifications TableValue of the data\u2022The data describes the meteorological and topographical conditions of the resort islands in the South China Sea\u2022This data contains key information for renewable energy assessments for resort islands in the South China Sea.\u2022This data can be used for other research fields that involve the usage of solar radiation, wind speed, rainfall and evaporation data.\u2022The topographic map data is valuable for determining the potential run-of-river hydropower sites in many resort islands in the South China Sea.1The data consists of meteorological and topographical data for resorts islands in the South China Sea.The selected resort islands location is shown in 2The meteorological data for the resort islands was obtained from the Malaysia Meteorological Department (MMS) and also the NASA Prediction of Worldwide Energy Resource (NASA POWER) database. If there is no meteorological station on the island, the meteorological data was collected from the nearest available meteorological station. The nearest station information\u05f3s also included in this article The topographic map obtained from Department of Survey and Mapping Malaysia (DSMM) was used for the study. The map is classified and must be used within the DSMM map library. The characteristic of the map used for this research is a Malaysian topography map (Series L 7030) with 1:50,000 scales and a contour interval of 20\u00a0m. The physiographic characteristics were extracted from the map for prediction of run-of-river hydropower potential sites. The important information extracted and analyzed from the map were: the name of streams and catchment areas, latitude and longitude of the location, lowest and highest elevation, the terrain and river profile, possible intake, diversion to fore bay, power house elevation, and the estimation of available head and catchment areas.Three key factors were considered for selecting catchment areas from a topographic map that were suitable for harnessing hydropower. The key factors were energy demand, accessibility and river profile. The catchment areas topographies were studied for determining the appropriate elevation for head and stream diversion. The river profile, which is the river\u05f3s tributaries and gradient, was considered for finding the availability of water resources and river flow. Based on the hydropower manual and guides"} +{"text": "In the Funding section, the first grant number from the Japan Society for the Promotion of Science is listed incorrectly. The correct number for the first grant is: Grant-in-Aid for Young Scientist (B) 25780404."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This project was supported by the President's Emergency Program For AIDS Relief and the NIH Office of AIDS Research, through NIH Grant Number U01 AI100015 to author SR at Boston University, from the National Institute of Allergy and Infectious Diseases (NIAID). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIAID. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There is an error in the \u201cAcknowledgements\u201d and \u201cFinancial Disclosure\u201d statements. Please refer to the correct statements below:Acknowledgments:We thank the more than 500 lay botanists who contributed to the Japanese Red Data Book project directed by the Japanese Society for Plant Systematics. We also thank Kazuo Somiya, of the Ministry of the Environment of Japan, and Yasuhiro Ibaragi, Masato Nagatsu, and Kaori Sato, of the Japan Wildlife Research Center, for their help and encouragement during the project. This work was supported by theMinistry of the Environment of Japan and the Japanese Society for Plant Systematics.Financial Disclosure:This work was supported in part by the Environment Research and Technology Development fund (S9-1) of the Ministry of the Environment, Japan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The Funding Statement is missing an acknowledgment to the Advanced Molecular Detection Initiative at the CDC. The correct Funding Statement is provided here:ET is supported by an American Society for Microbiology (ASM) and Centers for Disease Control and Prevention Postdoctoral Fellowship. We acknowledge support from the Advanced Molecular Detection Initiative at the CDC, the Atlanta Research and Education Foundation, Atlanta, GA, and the CDC Antimicrobial Resistance Working Group. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The following information is missing from the Funding section: Dr. Tarr is also supported by the National Institutes of Health [5P30 DK052574 (DDRCC)]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This work and survey components were supported by the High Impact Research Grant (UM.C/625/1/HIR/162) from the Ministry of Higher Education and University Malaya Research Grant (RP004A-SUS). Studies design and data collections were done by the academic staff at the Institute of Biological Science, University Malaya, Kuala Lumpur, Malaysia. Analyses undertaken and the decision to prepare and publish this manuscript were made by Institute staff in conjunction with the Chemistry Department of University Malaya."} +{"text": "There are errors in the Funding section. The correct funding information is as follows:This work was supported by the China 973 Project (Grant No. 2009CB941701) and National Natural Science Foundation of China (Grant No. 31071316) to XL. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This study was supported by the Institute for Clinical Evaluative Sciences (ICES) and Public Health Ontario (PHO), which are funded by annual grants from the Ontario Ministry of Health and Long-Term Care (MOHLTC). The opinions, results and conclusions reported in this paper are those of the authors and are independent from the funding sources. No endorsement by ICES, PHO, or the Ontario MOHLTC is intended or should be inferred.The images for figures Figs There was information omitted from the Acknowledgements. The complete Acknowledgements is as follows: These datasets were linked using unique encoded identifiers and analyzed at ICES. Parts of this material are based on data and information compiled and provided by the Canadian Institute for Health Information (CIHI). However, the analyses, conclusions, opinions and statements expressed herein are those of the author, and not necessarily those of CIHI. The authors wish to acknowledge the assistance of Danijela Draganic, Steven Janovsky and Lennon Li, Public Health Ontario, for assistance with preparation of figures for publication."} +{"text": "UK military personnel injured overseas are repatriated to the Royal Centre for Defence Medicine (RCDM) based at the Queen Elizabeth Hospital Birmingham (QEHB) in Birmingham UK. We report the demographics and outcomes of military patients treated on the ICU at RCDM using data from the Intensive Care National Audit and Research Centre over a 6.5-year period.Data on 570 admissions of 527 patients to the ICU at RCDM/ QEHB were analysed by ICNARC using standard methodology.Some physiology and CCMDS data were missing for 175 patients. Age, sex and mortality are described in Table The data on resource utilisation for this group of patients may inform planning of critical care support for military operations overseas."} +{"text": "The contents of this manuscript are the responsibility of IAVI and co-authors and do not necessarily reflect the views of USAID or the United States Government.There are errors in the Funding section. The correct funding statement is as follows: This work was made possible by generous support from many donors including; The training Health Researchers into Vocational Excellence in East Africa Project (THRiVE), Grant Number 087540 of Welcome trust, UK, and the Canada-Africa Prevention Trials Network (CAPTN) Grant Number 1063357\u2013001. It was also funded in part by IAVI and made possible by the generous support of the United States Agency for International Development (USAID) and other donors. The full list of IAVI donors is available at"} +{"text": "There is an error in the Funding section. The correct funding information is as follows: The study was funded by National Natural Science Foundation of China (No. 51378226). It had an important role in study design, data collection and analysis."} +{"text": "There are errors in the Competing Interests section. The correct competing interests information is as follows: This study was supported in part by the Agency for Science Technology and Research (A*STAR) Singapore. This does not alter our adherence to PLOS ONE policies on sharing data and materials."} +{"text": "Mental Health and Deafness. At the World Congress of Mental Health and Deafness held in 2014, a prize for the best poster was established in his name.Nick Kitson, who died suddenly at the age of 64 while on holiday on 24 February 2014, was a pioneer and national expert in the psychiatric disorders of deaf people. He was the Founding Chair of the British Society for Mental Health and Deafness and the longest serving president of the European Society for Mental Health and Deafness (1990-1997), remaining honorary member and honorary vice president of each, respectively. He was clinical advisor to the Towards Equity and Access Implementation Panel in 2006/2007 and \u2018responsible owner\u2019 of a National Deaf Mental Health Commissioning Project. He remained a trustee of and medical advisor to the Sign health charity and awarded the title of Pioneer. He was joint editor of the standard introductory textbook Nick completed his medical training at St Bartholomew's Hospital in 1975. He was appointed consultant psychiatrist at Springfield University Hospital and honorary senior lecturer at St George's Medical School in 1984. He was clinical director for the Specialist Mental Health Services for the Deaf Community at Springfield and medical director of Pathfinder NHS Trust from 1994 to 1998. When he became responsible for the psychiatric care of people with deafness, his health authority gave him 9 months to learn sign language. He visited psychiatric units for deaf people in America and realised the importance of employing deaf people fluent in sign language. At the time he wrote about sign language: \u2018It is a very sophisticated language capable of expressing everything you can say in English.\u2019 The service for deaf people that he led and developed for 18 years, with his wife Karen, covered the southern half and then a third of England. He became a Member of the Royal College of Psychiatrists in 1980 and was elected a Fellow in 1996.While working as a consultant, he trained in short-term dynamic therapy in 1984 and group analytic psychotherapy in 1989. He became an associate member of the London Centre for Psychotherapy (British Psychotherapy Foundation) in 1989 and a Full Member in 1996, supervising short-term dynamic psychotherapy at the Tavistock and latterly at the London Centre for Psychotherapy. Psychotherapy was always an integral part of his clinical practice. He provided strong support for Jane Douglas, the first profoundly deaf person to train as a psychoanalytic psychotherapist.Knowing the West Country well, Nick left London and moved to Cornwall in 2002. He initially worked as a community and in-patient psychiatrist covering the St Austell area but, wishing to reduce his workload owing to ill health, he became part-time consultant to the psychiatric intensive care unit (PICU). The service was awarded the National PICU of the Year in 2005. Formally retiring in 2009, he continued as his own locum for the PICU, regularly sailing and taking trips abroad with Karen and his family.Nick was a hugely respected consultant psychiatrist. His colleagues frequently sought his advice in difficult circumstances, and the advice he gave was wise and supportive. He was down-to-earth and sensible; he never pretended he knew the answer when he did not, but his advice was even sounder in these circumstances. He continued to use his therapeutic skills after retirement and remained enthusiastic and determined to help the most challenging and difficult patients.He will be sorely missed by his colleagues but his wise leadership will continue to be influential. He is survived by Karen, his two daughters and his grandchildren."} +{"text": "She is with the European Programme for Intervention Epidemiology Training (EPIET) at the European Centre for Disease Prevention and Control, Stockholm, Sweden. The article has been corrected online ("} +{"text": "The role of medical schools is in a process of change. The World Health Organization has declared that they can no longer be ivory towers whose primary focus is the production of specialist physicians and cutting edge laboratory research. They must also be socially accountable and direct their activities towards meeting the priority health concerns of the areas they serve. The agenda must be set in partnership with stakeholders including governments, health care organisations and the public.The concept of social accountability has particular resonance for the Bar Ilan Faculty of Medicine in the Galilee, Israel\u2019s newest medical school, which was established with a purpose of reducing health inequities in the Region. As a way of exploring and understanding the issues, discussions were held with international experts in the field who visited the Galilee. A symposium involving representatives from other medical schools in Israel was also held to extend the discourse. Deliberations that took place are reported here.The meaning of social accountability was discussed, and how it could be achieved. Three forms of action were the principal foci \u2013 augmentation of the medical curriculum, direct action through community engagement and political advocacy. A platform was set for taking the social accountability agenda forward, with the hope that it will impact on health inequalities in Israel and contribute to discussions elsewhere. Closing the Gap in a Generation[, it reported that many of the differences in health between and within countries stem from the social environment where people are born, live and age. The Commission recommended a two-pronged approach to redressing health inequities \u2013 improving people\u2019s daily living conditions and tackling inequitable distribution of power, money and resources.In 2008 the World Health Organisation (WHO) published a report written by its Commission on Social Determinants of Health. Entitled eneration, it repoAlthough medical professionals can only have a limited role in implementing these recommendations, they nonetheless have a clear responsibility to address health inequities in the course of their work. This challenge was taken up by pioneering medical schools, many of whom were new schools. They were established with a paradigm shift - embedding improvement in community health with innovations in health professional education, with delivery and health policy at the core of their business. These schools engaged initially in establishing a network of community orientated medical schools, of which Ben Gurion University of the Negev was one, and evolved into an official relationship with the WHO as The NetWork.The concept was taken further at a groundbreaking international meeting in South Africa in 2009 bringing together representatives of 130 organisations and individual experts from around the world with responsibility for health education, professional regulation and policy making to produce a Global Consensus for Social Accountability of Medical Schools (GCSA 2010),5.Closing the gap in a generation[Global Consensus[eneration and the onsensus have speonsensus.This discussion document reflects some of the ideas and discussion that took place. Issues that were explored included: what is meant by social accountability; how it can be achieved; the role of medical education, direct action and advocacy; and how we can measure success over time.The World Health Organisation defines social accountability as the obligation for medical schools to direct their activities towards meeting priority health concerns that are agreed in partnership with stakeholders nationally and locally. In many ways it is the next stage in the service, research and education revolution started several decades ago by the Community Oriented Primary Care movement, developsocially responsible when they teach about health inequalities and the impact that social determinants have on health. Social responsiveness goes a step further as it involves taking action in response to societal or local community health needs. Abstracts submitted to the symposium originally implying shared responsibility for proper social and moral conduct in the community, but used in more recent centuries to connote mutual responsibility for social welfare.\u201cTiqqun \u2019olam\u201d was translated in the title of Jonathan Sacks\u2019 book as \u201cTo heal a fractured world \u2013 the ethics of responsibility\u201d .None of the authors have declared competing interests.All the authors made substantial contributions to the discussions underpinning this paper. The manuscript was drafted by MCJR and the remaining authors contributed critical revisions and gave final approval of the version submitted to IJHPR. They all agree to be accountable for all aspects of the work and in ensuring that questions related to the accuracy or integrity of any part are appropriately investigated and resolved. All authors read and approved the final manuscript.Professor Mary Rudolf is the lead for Public Health at the Bar Ilan University Faculty of Medicine in the Galilee since 2012, and visiting Professor of Child Health at the University of Leeds, UK. Areas of special interest include childhood obesity, paediatric education and social accountability of medical schools. Professor Shmuel Reis is a Family Physician in a rural health center, head of the Faculty Development Unit and the Clinical Skills Course director of the Bar Ilan University Faculty of Medicine in the Galilee. Professor Trevor Gibbs is an Independent and WHO Consultant in Medical Education, Primary Care and Adolescent Health. He also holds the position of Development officer for AMEE. His main areas of interest are in International medical curricula, Competency-Based Education, Social Accountability and Nutrition and Health. Professor Deborah Murdoch-Eaton is Dean of Medical Education, at The Medical School, University of Sheffield, UK. Her academic interests focus on Global Health, developing students' potential and individuality, embedding Social Accountability within medical education, and role of feedback in the development of learning skills. Professor David Stone is Emeritus Professor of Paediatric Epidemiology at the University of Glasgow, UK. Previous positions included Senior Lecturer in Epidemiology at Ben Gurion University of the Negev, Israel and Senior Medical Advisor to the Scottish Government\u2019s Chief Medical Officer. Dr Michael Grady is an expert adviser to WHO and special adviser to the UK House of Commons Community and Local Government Select Committee. He advises Local authorities and municipalities nationally and internationally on implementation on action on the social determinants of health. Dr Anita Berlin is an inner London General Practitioner and a senior lecturer in primary care at University College London. She is curriculum lead for the new Social Determinants of Health Module and for Patient and Public Participation. Prof Mitch Blair is Consultant Paediatrician and Reader in Child Public Health Imperial College London and Northwick Park Hospital. He is undergraduate paediatrics course lead for Imperial medical students and has a strong interest in teaching Child Rights and Health Inequalities. Dr Jumanah Essa-Hadad has a PhD in Public Health from the University of Haifa. Research interests include web-based health education and health of minority groups and disadvantaged populations. Her background includes working with NGO's and civil society organizations at the grassroots level to promote public health programs. Ms Sivan Shohat holds a Master's degree in Sociology specializing in health care policy and the role healthcare organizations have in reducing health and healthcare disparities. Professor Michael Weingarten is Associate Dean for Medical Education at the Bar Ilan University Faculty of Medicine in the Galilee. A family practitioner by background, he has a special interest in medical ethics."} +{"text": "The following information is missing from the Funding section: grant number LD13046 from the Ministry of Education, Youth and Sports of the Czech Republic and the acknowledgment of funding from the EU COST action ES1103.www.daad.de) providing travel grants for VR and MS, from the Ministry of Education, Youth and Sports of the Czech Republic , from the Grant Agency of the Czech Republic , and from the EU COST action ES1103, which provided funding for the author (AC) to attend a workshop.The complete, correct Funding statement is: This project was funded by a grant from the German Academic Exchange Service (DAAD,"} +{"text": "In the Materials and Methods, an Institutional Animal Care And Use Committee (IACUC) protocol number was incorrectly cited. The authors wish to correct the record by reporting that there was no University of Texas at Arlington (UTA) IACUC protocol number for this research because the research was not conducted at UTA, with the exception of the dissection of the python, which took place without UTA IACUC approval. The corrected text and additional details and clarification are provided below for the tissue sample from the Burmese python, as well as the samples from two birds, the Gunnison Sage-grouse and Clark's Nutcracker.Python molurus bivittatus was obtained by the UTA Amphibian and Reptile Diversity Research Center from a commercial breeder in August of 2007, immediately euthanized using a chloretone solution, and tissues preserved following animal protocols outlined in the Guidelines for Use of Live Amphibians and Reptiles in Field Research, established jointly by the American Society for Ichthyologists and Herpetologists, Herpetologist's League, and the Society for the Study of Amphibians and Reptiles . Liver tissue from the python (snap-frozen in liquid nitrogen and stored at -80\u00b0C) was used as a source for genomic DNA.\"All specimens used in this study were obtained from collaborators and not collected by the authors. There was no University of Texas at Arlington (UTA) IACUC protocol number for this research as the research was not conducted at UTA, with the exception of the dissection of the python, which took place without UTA IACUC approval. A single The Gunnison Sage-grouse was trapped and blood was sampled by personnel from the Colorado Division of Wildlife. The trapping method and blood sampling was approved through their Animal Care and Use Committee; however, at the time, no permit was required to trap and sample the bird (the bird was still being actively hunted at the time when the sample was taken). There was no number issued in terms of an IACUC.The tissue from the Clark's Nutcracker was provided from a dead carcass of a bird that died of natural causes in the lab of Russell Balda at Northern Arizona University after more than a decade as an experimental bird for memory studies. The bird was originally caught in Logan Canyon in the 1990s by Stephen Vander Wall (University of Nevada Reno), under the IACUC protocol number 00\u2013006 Russell Benford, Ph.D., Department of Biological Sciences, Northern Arizona University."} +{"text": "In the Funding section, the grant number from the funder the National Natural Science Foundation of China is listed incorrectly. The correct grant number is: 31072055. The complete, correct funding information is as follows: This work was financially supported by the National Natural Science Foundation of China (Project no. 31072055) and The Ministry of Science and Technology of China (Project no. 2011BAD26B03-4). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There is an error in the Funding section of the published article. The Funding section should read: This study was supported by a grant of the Jeju National University Research Fund (2011). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In the Acknowledgements section, the following information is missing: All data in this paper were provided by the TEAM Network, a partnership between Conservation International, The Missouri Botanical Garden, The Smithsonian Institution and The Wildlife Conservation Society."} +{"text": "The Acknowledgments section is incorrectly reported. The correct Acknowledgments section should read: We want to thank Dr. Rabie Mohammed Algdgad and Dr. Ahmed Bazza, Tripoli Medical Center for the contribution in collecting of data, and Prof. Michael Eddleston, Edinburgh, Scotland for his invaluable input and critical review of the manuscript. Study expenses were supported from the regular operation budgets of the MSF Kenya and Libyan missions as well as Oslo University Hospital, but had no role in the writing of the report as such."} +{"text": "India has the highest number of medical colleges in the world and subsequently the higher number of medical teachers. There is a dire need of adopting a systematic approach to faculty development to enhance quality education to meet health challenges for 21st Century. This manuscript provides a landscape of faculty development programs in India, identifying gaps and opportunities for reforms in faculty development. Conventionally, FDPs are organized by medical colleges and universities through Basic Courses and Advanced Courses focusing on pedagogy. Medical Council of India is facilitating FDPs through 18 selected regional centers to enable medical teachers to avail modern education technology for teaching from July 2009. Foundation for Advancement of International Medical Education and Research has three Regional Institutes in India. Recommendations include the need for formulating a national strategy for faculty development to not only enhance the quantity of medical teachers but also the quality of medical education; providing support for Departments of Medical Education/Regional Centers in terms of finance and staffing and incorporation of teaching skills in postgraduate training. Distance learning courses focusing on educational leadership and pedagogy for medical teachers can be an option to reach a wider audience. FDPs can be an asset in recruiting and retaining teachers as they offer valued professional development opportunities. The unprecedented growth of medical institutions in India in the past two decades, almost doubling in strength, has led to a shortage of teachers and created a quality challenge for medical education, Faculty Development Programs aim to improve the quality of medical education by training and sensitizing teachers about new concepts in teaching and assessment methods; develop knowledge and clinical skills required for performing the role of competent and effective teachers, administrators, researchers and mentors; assist clinicians to acquire competency in communication and behavioral skills and update knowledge using modern information and research methodology tools. HoweverConventionally, the FDPs in India have been organized by medical colleges and universities through Basic Courses and Advanced Courses on pedagogy envisaged to improve the quality of medical education by training the medical teachers. The purpose of the Basic Courses in Medical Education Technologies (MET) is to provide the basic knowledge, skills and eventually change the attitude of the faculty in medical colleges which the faculty can implement in their day to day practice in different areas of teaching and assessment . MCI, in 2014, started Advanced Courses in medical education with the aim to develop educational practitioners who can lead informative and instructional and educational changes in their institutions and thereby making the medical education responsive to the health needs of the society. . Bhore Committee recognized the need for training of medical teachers in as early as 1946 and made recommendations for major changes in medical education which included three months of training in preventive and social medicine to prepare \u201csocial physicians\u201d. Fifteen years later, Mudaliar Committee in 1961, re-emphasized the need for the \u201csocial physician\u201d. Patel Report, in 1971, described a \u201cbasic doctor\u201d of modern medicine who would be central to the delivery of primary healthcare and trained through a five-and-a-half years of university education. In 1974, Srivastav Committee advocated the set up for establishment of Medical and Health Education Commission for planning and implementing the reforms needed in health and medical education on the lines of University Grants Commission. An \u201cExpert Committee for Health Manpower Planning, Production and Management\u201d established in 1985 and known as Bajaj Committee, further urged for a formulation of national medical & health education policy and the nodal agency was University of Illinois, United States. The IRTTC trained faculty from six Regional Teacher Training Centers (RTTCs). Two RTTCs were established in South-East Asia, one in Sri Lanka and one in Thailand supported by WHO. Government of India, constituted a Working Group on Continued Medical Education in 1974 which recommended National Teacher Training Centre (NTTC). The first NTTC was established in 1975 at JIPMER, Pondicherry and offered the National Courses on Educational Science for Teachers of Health Professions that are held twice a year. In March 2014, NTTC, JIPMER, Pondicherry organized its 69ew Delhi , 9. The ew Delhi .Consortium of Medical Institutions for Reform of Medical Education also played a crucial role in advancing the FDPs in India from 1989 to 1995. Four medical institutes, viz, All India Institute of Medical Sciences, New Delhi; Christian Medical College, Vellore; JIPMER, Pondicherry and IMS-BHU, Varanasi and the Department of Medical Education, College of Medicine at Chicago, University of Illinois, formed a consortium. Later this consortium was expanded to 16 colleges. The consortium contributed to curriculum development, built consensus and classified essential skills, into ''must know'' and ''good to know'' categories.K.L. Wig Centre for Medical Education and Technology was another strong catalyst in promoting faculty development in India. It was established at All India Institute of Medical Sciences, New Delhi in 1989-90. Rajiv Gandhi University of Health Sciences, Karnataka, Bangalore and The Tamil Nadu Dr. M.G.R. Medical University has a separate department of curriculum development and promotes Health Professionals' Education and aims to develop need based curricula for the medical and paramedical education. The Indian Journal of Medical Education was launched as an official publication of the Indian Association for the Advancement of Medical Education. The Department of Medical Education of K.M.C. Manipal was established in 1985 to bring about improvements in medical education and health care and regularly holds Teaching-Learning Workshop and Student Evaluation Workshops. The National Knowledge Commission was established by the Government of India in 2005 to recommend and undertake reforms in order to make India a knowledge-based economy and society. An important constituent of the National Knowledge Commission functions was professional education, particularly education in the field of medical sciences .MCI has taken a major initiative of conducting FDPs in India by making regulations on Graduate Medical Education in 1997 which made it mandatory for all medical colleges to establish Department of Medical Education. MCI has also selected 18 Nodal Centers, since July 2009. These centers are located at institutions which have trained manpower in Medical Education Technologies (MET) . All theMore than 40000 faculty are estimated to be working in 387 medical colleges based on the MCI norms and 12141 teachers have been oriented by MCI under FDPs through 429 workshops. The Nodal Centers of MCI have trained 5946 medical teachers in 186 workshops since its inception. And through the Medical Education Unit/ Department, 243 workshops have been conducted and 6195 medical teachers have been trained is also assisting faculty development in India. The FAIMER Institute, started in 2001, is a two year fellowship focused on educational leadership and methodology. There are seven Regional Institutes in the world out of which three are in India, one in Brazil, one in Southern Africa, one in China and one in Colombia. In India, the first regional institute, Seth G S Medical College was started in July 2005 in Mumbai, India, The second CMCL-FAIMER Regional Institute, fellowship started in January 2006 based at the\u00a0Christian Medical College at Ludhiana, Punjab. It is based at\u00a0PSG Institute of Medical Sciences and Research\u00a0in Coimbatore in southern India.\u00a0 These three Regional Institutes are open to South Asia health profession educators and sixteen fellows are accepted each year. The proEducation for Health\u201d of The Network: Towards Unity for Health, a global consortium of health professions schools.Various training programs incorporating innovative models for faculty development are being conducted by various academies, associations, consortia and the deemed universities like Indian Association for Advancement of Medical Education, Consortium of Medical Institutions for Reform in Medical Education, Health Science University Initiatives, Consortium of Health Science Universities and Indian Academy of Health Profession Education. Apart from the MCI Nodal Centers, many other health sciences universities have initiated courses and programs in FDPs. Maharashtra University of Health Sciences (MUHS), Nasik, Maharashtra has established the Institute of Medical Education Technology & Teachers\u2019 Training and has completed 12 trainings on \u201cAdvanced Certificate Course in Health Sciences Education Technology\u201d till January 2014. It aims to impart advanced educational skills to teachers in positions of academic responsibility in their own institutions. This certificate course is of six months duration in which there is a seven days contact session consisting of full day sessions followed by a six month educational innovation project(Datta Meghe Institute of Medical Sciences, Wardha established the Department of Health Professional Education in 2009 and offers an Advanced Course in Medical Education recognized by MCI and 30% of the faculty of Datta Meghe Institute of Medical Sciences have been trained in Advanced Courses in Medical Education. DMIMS is also a MCI Regional Centre and has trained more than 1000 medical teachers from more than 15 medical colleges.\u00a0 The institute also has six FAIMER faculty and three National Faculty for Advanced Courses in Medical Education recognized by MCI. South East Asia Regional Association for Medical Education of the World Federation for Medical Education is also supporting in this endeavor by promoting partnerships and linkages with various local and regional associations to collaborate for medical education under the umbrella of World Federation for Medical Education. Though several initiatives are being undertaken by various agencies to build the pedagogy skills of medical teachers, the flipside is that the number of workshops, trainings and courses are too meager to address the huge need. In near future, in addition to the existing medical colleges, HLEG on UHC has proposed establishing new 187 medical colleges by 2022. These mAlthough the MCI Basic Course has been made compulsory for medical teachers to the level of professor and teacher administrators, it is also being recommended that the Medical Education Units in all medical colleges should train all existing medical teachers in Basic Course and conduct the Basic Course for newly inducted faculty twice a year. MCI UndNodal faculty development centers, Medical Education Units and Centers of Regional FAIMER Institutes in India have been able to do the capacity building and ultimately helping to increase the ability of systems to function on their own to meet local needs . MCI hasA big lacunae at present is in terms of creating and developing an educational leadership. Programs need to be designed to educate policy makers and update them about recent advances in medical education worldwide. These include engaging people\u2019s moral purposes, building capacity to generate forces for change, understanding the change process, developing the learning culture and the culture of evaluation and fostering development at all possible levels. This report highlights the need for strengthening faculty development as a vehicle for ensuring quality in medical education. There is evidence, in most countries, educators of health professionals are insufficiently prepared as teachers and trainers, even though their clinical knowledge and skills may be good. The success of educational reforms ultimately lies with the individual instructors and their capacity, individually and collectively to execute and implement some novel ways in teaching and training the future cadre of doctors and more importantly with India progressively becoming a new global hub of education. Faculty development efforts should empower the medical teachers and keep the passion in teaching going so that the lifelong learning never ceases."} +{"text": "The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.A grant number is missing in the Funding section. The correct funding information is as follows: The project is supported by the National Institutes of Health grants 5R01GM080646-09 and HHSN272201000053C ("} +{"text": "It has come to our attention that there was an omission in the Acknowledgements section in this article . The AckThe authors are grateful to the study participants, to the doctors and nurses at the Oxford Vaccine Group for assisting with sample collection, and to the National Institute for Health Research Clinical Research Network. The authors thank Craig Waugh for help with cell sorting and the High-Throughput Genomics Group at the Wellcome Trust Centre for Human Genetics (subsidized by Wellcome Trust grant reference 090532/Z/09/Z) for the generation of sequencing data. Purified HBsAg was provided by GlaxoSmithKline Biologicals SA, and conjugated to APC by Miltenyi Biotec."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This study was supported by the National Cancer Institute of the National Institutes of Health under award number K08CA155035 and the Melanoma Research Alliance. The authors are also grateful to Timothy Dattels for his generous support. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There is an error in the Correction published on August 19, 2014. The affiliation for Samuel Ginsberg is incorrect. Samuel Ginsberg is not affiliated with #3, please refer to his correct affiliation here: Department of Electrical Engineering, University of Cape Town."} +{"text": "There is an omission in the Funding Statement. The correct funding information is as follows: This work was supported by Council of Scientific and Industrial Research (New Delhi): Network Projects BSC 0114 and BSC 0120. MG, KR and DDM are thankful to CSIR for fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: The present study was funded by the Wellcome Trust and the Medical Research Council Programme , and the R24 Alcohol Research Resource Award grant (R24 AA015512) from NIAAA. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In the Funding section, the name of the first funder is incorrect. The correct name is: Medical Research Scotland. The correct funding information is: This work was supported by Medical Research Scotland [grant 354FRG to AJC] and the Medical Research Council [grant G1100357 to RAA]. J.H. is supported by a University of Edinburgh Darwin Scholarship and K.S. is the recipient of a Society for Reproduction and Fertility Summer Vacation Scholarship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There is a missing funder in the Funding section. Please refer to the complete funding statement here:We thank Karen Frutchey and Raymond Clarke of the Pacific Islands Regional Office, National Marine Fisheries Service (NMFS) and National Oceanic and Atmospheric Administration (NOAA) who provided key funding towards this project. The funders were base funding and add-on funding from the United States government to NOAA and had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The grant funding from J. Juliano is missing from the original article. The correct funding statement is as follows:This study was funded by the Institute of General Medical Science\u00a0(R01 GM089932 to AFR) and Institute of Allergy and Infectious Disease of the National Institutes of Health. The funders had no role in study design, data collection and analysis, decision to publish or the preparation of the manuscript."} +{"text": "Alan Wilton is not included in the author byline. He should be listed as the 23rd author and affiliated with School of Biotechnology & Biomolecular Sciences, Faculty of Science, The University of New South Wales, Sydney, NSW 2052, Australia. The contributions of this author are as follows: sample collection.The following sentence should also be included in the acknowledgements. \"The authors also wish to thank the Australian Native Dog Conservation Society and the Dingo Sanctuary Bargo for generously providing the dingo sample used in this study.\""} +{"text": "The Weight Disorder Unit at Flinders Medical Centre was established in 1977 and in June 2014, the service was expanded under a rebranded name, the South Australian Statewide Eating Disorder Service (SEDS). Now, besides its inpatient unit, SEDS also has outpatient services and an eating disorder Day Program. This is the first Day Program in South Australia and besides providing nutritional support, the Day Program also focuses on helping participants to challenge disordered eating behaviours and thought patterns through group-based therapeutic interventions. To be admitted into the SEDS Day Program, clients need to be at least 15 years of age, have a Body Mass Index (BMI) of 15 and above and be medically stable. In order to gain a better understanding of the profile of clients that get admitted to day programs and the effectiveness of the Day Program, we plan to present the clinical profile of the participants and information collected from the SEDS Day Program. Information presented will include sociodemographic and medical information, information collected from clients' psychological assessments pre, post and 3 months follow-up after completion of the Day Program, and the retention rate of the Day Program participants."} +{"text": "The funders had no role in study design,data collection and analysis, decision to publish, or preparation of the manuscript.\" There are multiple errors throughout the Funding section. The complete Funding sectionshould read: \"This work was supported by National Natural Science Foundation of China(30972239 and 31130054,"} +{"text": "There is an error in the Funding section. The correct funding information is as follows: The research was supported by National Institute of Health grant R21HL117652 and grants from University of Minnesota Academic Health Center. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The publisher apologizes for the error."} +{"text": "In the Funding section, the grant number for the Ministry of Education, People\u2019s Republic of China grant is incorrect. Please refer to the correct Funding section here.This research was financially supported by The National Natural Science Foundation as a key project (grant No. 21037002), and by the Ministry of Education, People\u2019s Republic of China as an innovative research team project (grant No. IRT13024). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The following information is missing from the Funding section: Publication of this article was funded in part by the George Mason University Libraries Open Access Publishing Fund. Please refer to the complete funding statement here.http://ragnarsoderbergsstiftelse.se/, grant number: E56-10), Kungliga VetenskapsAkademin and the Lab for Economics Applications and Policy at Harvard . Publication of this article was funded in part by the George Mason University Libraries Open Access Publishing Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Financial support for this project is gratefully acknowledged from: the Ragnar S\u00f6derberg Foundation ("} +{"text": "There are errors in the Funding section. The correct funding information is as follows: Funding was provided in part by NIH/NIAID U01 AI075526 and in part by the Bill & Melinda Gates Foundation OPP1066140. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Natural Resources Defense Council was omitted as a funder for this study. Please refer to the correct funding statement below:Dr. Hess's work on this project was via a consultancy arrangement with the Natural Resources Defense Council (NRDC) funded by the Climate and Knowledge Development Network (CDKN). This work was funded by a grant from the Climate Knowledge Development Network. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The following information is missing from the Funding section: DB acknowledges NSF grant 1354793. The complete, correct funding information is as follows: The genetics and biogeographic research in this paper was funded by an Australian Research Council grant ; the linguistic research was supported by a grant from the Kimberley Foundation Australia (awarded to McConvell), a US NSF Grant , and an AIATSIS grant . DB acknowledges NSF grant 1354793."} +{"text": "There is an error in the first sentence of the Funding section. The first sentence should read: This work was partially supported by grants from the National Institute of Allergy and Infectious Diseases (NIAID) and partially from Integrated BioTherapeutics, Inc. (IBT) internal R&D research fund."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: Guoxin Zhang was funded by National Natural Science Foundation of China (No. 81072032 and No. 81270476) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (JX10231801); Xiaoying Zhou was funded by Jiangsu postgraduate scientific research and innovation projects (CXZZ13_0574). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Marc Arbyn was supported by the seventh framework program of DG Research of the European Commission, through the COHEAHR Network (grant No 603019). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There are errors in the Funding section of the published article. The correct funding information is as follows: Swedish Cancer Foundation URL:"} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This work was supported by the Korea Polar Research Institute (Grant PE14070). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "MK wishes to declare the following competing interest, which was omitted from the original article: MK is a founder of the Dog Aging Project, a University of Washington part-funded project that also solicits public donations through the University of Washington Foundation."} +{"text": "The following information is missing from the Funding section: This work was supported by National Institutes of Health grant HL114669.The complete, correct Funding statement is as follows: This work was supported by the Nebraska Research Initiative Grant, and the National Institutes of Health (HL114669). CM is a recipient of a postdoctoral research fellowship grant awarded by the Myocarditis Foundation, NJ."} +{"text": "The importance of scientific evidence to guide nutrition policy and programme design is well established.In an effort to address these evidence gaps, the Global Nutrition Report of 2014While in most countries governments regulate food fortification \u2013 i.e. what can be fortified, with which nutrients and at what levels \u2013 monitoring of compliance and enforcement of regulations needs to be strengthened considerably in many countries.The big question remains: how to garner the resources and political commitment needed for the generation and use of evidence for programme decision-making in nutrition broadly and food fortification specifically? This question was high on the agenda at the first Global Summit on Food Fortification in Arusha, United Republic of Tanzania from 9 to 11 September 2015. Government delegations from 29 countries from Africa, Asia and Central and South America, and experts and representatives from donors, United Nations agencies, nongovernmental organizations working in food fortification and the private sector, met to discuss and debate the state of food fortification in their countries and globally. The Summit declaration"} +{"text": "There is an error in the name of the funding organization, Hans und Gerti Fischer Foundation. Please refer to the correct funding statement below:This research was supported by the Dr. Heinz-Horst Deichmann Foundation and the Hans und Gertie Fischer Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Dr. John P. Hart is not included as the Academic Editor of this article. He should be listed as the Academic Editor and is affiliated with the New York State Museum, UNITED STATES."} +{"text": "The Joint Research Centre (JRC) is a Directorate-General (DG) of the European Commission (EC), it was established in 1957 and its mission is to provide independent, evidence-based scientific and technical support throughout the EU policy cycle. Health is a key European policy area and research on Rare Diseases (RD) is identified as a priority in both the Commission Communication on RD: Europe's challenges COMM (2008) 679 final and the Council Recommendation on an action in the field of RD (2009/C 151/02).The specificities of RD \u2013 limited number of patients and scarcity of information \u2013 single them out as a unique domain for which an action at European level has high potential added value. For this reason and in the context of developing a long-term EU strategy in the area of health information including RD data collection, DG Health and Consumers (SANCO) and the JRC started an initiative aimed to improve the sustainability of the results achieved over the past years. Taking into account that the current fragmentation of data sources is a key obstacle to steering EU policy and to performing high quality research and in the end to advancing knowledge on RD, the two DGs agreed on the development of the European Platform for RD registries. The main purpose of the Platform which will be established at the JRC is to support the existing registries by promoting their interoperability and accessibility, by improving RD data comparability and reliability. It will also support the development of new registries. This 'hub' will facilitate data analysis within and across many RD and across Europe and provide sound information on RD for policy, clinical trials and research. The Platform is intended to provide a central access point for information on RD patients registries for all stakeholders \u2013 health professionals, researchers, patients, public health authorities, industry, etc. Thus the benefits of collaboration and maximisation of limited resources are most obvious in a concerted action with the final aim of improving the quality of care and the quality of life for RD patients."} +{"text": "The following information is missing from the Competing Interests section: \"The views expressed are those of Dr. Hamrick\u2019s, and should not be attributed to the Economic Research Service or USDA. \"The correct competing interest information is as follows.Dr. Hamrick, one of the co-authors, is employed by the USDA Economic Research Service. The views expressed are those of Dr. Hamrick\u2019s, and should not be attributed to the Economic Research Service or USDA. There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to all PLOS ONE policies on sharing data and materials."} +{"text": "The outcome of different studies on the role of Zn & Mo in esophageal cancer (EC) is conflicting. Here, the levels of those elements in hair as well as food grain of two different ethnic populations across two continents have been studied to explore their role in EC. Two different ethnic populations are taken from (i) Eastern Cape, South Africa (RSA), an area of very high incidence of EC and (ii) West Bengal, India, an area of low incidence of that disease. Each ethnic population is divided into two groups: case and control . Hair samples from all groups and food grains from RSA and India are analyzed for Zn & Mo content. This study shows a strong correlation between reduced levels of those elements in hair and the development of EC in RSA (both Zn & Mo: p<0.0001), though it is only suggestive in Indian context (both Zn & Mo: p\u22650.05). Interestingly, control group of RSA shows significantly reduced level of those elements in hair even with respect to Indian case group (Zn: p<0.001 & Mo: p<0.00001). Food grain from RSA has significantly reduced level of those elements with respect to India (both Zn & Mo: p<0.0001). This deficiency of Zn & Mo in food grains can be correlated to the deficiency of those elements in hair of RSA population. The deficiency of Zn & Mo can be correlated to the development of EC. In South Africa, esophageal cancer (EC) is the third most common cancer in males but the incidence is moderately high among black males . WorldwiThe role of Zn & Mo has been studied by many investigators. Some studies from endemic area of EC reported that plasma zinc was significantly reduced in EC compared to age matched healthy controls . In ChinSeveral retrospective and prospective studies on humans and animals have shoSome studies have indicated that dietary deficiency of Zn is associated with the development of EC but the role of other trace elements like Mo are not conclusive . ConflicHowever, it is found that outcome of different studies are conflicting and the literature in this field are also scanty indeed. So, the role of trace elements (Zn & Mo) in the etiology of EC needs to be explored. Zn & Mo are deposited in hair and one of the sources of these elements is food grains consumed. In the present work the role of Zn & Mo in the etiology of EC has been studied by analyzing the amount of Zn & Mo both in food grain and hair.Epidemiological studies indicate that Eastern Cape, South Africa is one of the hot spot and West Bengal, India is one of the cold spot for EC. That is why Indian population from West Bengal has been taken for this study.The study included the following groups:Thirty EC patients are taken from the Surgical Department of Frere Hospital, Eastern Cape, South Africa. They are all above 18 years age, from black ethnical background and have squamous cell carcinoma of the esophagus. They have resided for more than 15 years in Eastern Cape, South AfricaThirty self volunteered people from the general population are taken. They are all above 18 years, from black ethnicity and do not have any history of EC. They have resided for more than 15 years in Eastern Cape, South Africa.Case group: Thirty EC patients are taken from the Surgical Department of CCWHRI Hosp, Kolkata, India. They are all above 18 years age, from Indian ethnical background and have squamous cell carcinoma of the esophagus. They have resided for more than 15 years in West Bengal, India.Control group: Thirty self volunteered people from the general population are taken. They are all above 18 years, from Indian ethnicity and do not have any history of EC. They have resided for more than 15 years in West Bengal, India.Inclusion criteria for all case groups:a)Both males and females are above18 years.b)Their staple dietary source is from local crops grown .c)The weight of all the subjects is above 36 kg.d)The patients are able to swallow at least a liquid diet.Control Group is taken from volunteered participation of the respective population of the respective study area.Inclusion criteria for all control Group:a)Physically and clinically normal adult above 18 years whose weight is above 36 kg.b)Their staple dietary source is from local crops grown .c)None of the subjects has a previous history of taking Zn & Mo supplementation in the previous two weeks.Food grain from residing places of the case and control group from Eastern Cape, South Africa as well as West Bengal, India are taken for evaluation of Zn & Mo content. The staple diet of the people in these areas is maize and rice for RSA and India respectively. Number of samples for food grain analysis is thirty for both RSA and India. Hair samples are collected from all subjects by a doctor and kept in a special container for Zn & Mo analysis.Zn & Mo are analyzed according to the standard methods of American Public Health Association . Both foOur results shows that EC patient of South African case group has significantly reduced level of Zn & Mo in hair with respect to the South African control group (both Zn & Mo: p<0.0001) ; Table 1Zn & Mo concentration of food grain in RSA and India shows a correlative pattern (both Zn & Mo: p<0.0001) ; Table 4All the statistical evaluations are indicating a deficiency of Zn & Mo in hair among RSA population and the deficiencies of Zn & Mo may play a distinct role for the development of EC in Eastern Cape, RSA (hot spot) and this is also supported by other studies ("} +{"text": "Unfortunately, the original version of this article containeThe authors gratefully acknowledge the funding of this research provided by the Ministry of Health and Consumers\u2019 Affairs - \u2013Spain (FIS Exp. PI13/01340 and FEDER funds) and the CHIR- Quebec Training Network in Perinatal Research (QTNPR). The study funders had no role in the study design, data analysis, data collection, data interpretation or the writing of the report. The views expressed are those of the authors and not necessarily of the funding bodies."} +{"text": "There is a grant number missing from the Funding Statement. Please see the corrected Funding Statement here.This work is funded by the National Institutes of Health, National Institute of Allergy and Infectious Diseases (AI118732) to MMK and the training grant (5T32AI007046) to CJA. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The following information is missing from the Funding section: JA received funding from Magnus Bergvall Foundation ("} +{"text": "The Alexander von Humboldt Foundation promotes academic cooperation between excellent scientists and scholars from abroad and from Germany. To this end, it grants more than 700 research fellowships and research awards annually. These allow scientists and scholars from all over the world to come to Germany to work on a research project they have chosen themselves together with a host and collaborative partner. Scientists or scholars from Germany can also profit from the support and carry out a research project abroad as a guest of one of well over 26,000 Humboldt Foundation alumni worldwide \u2013 the Humboldtians.Once a Humboldtian, always a Humboldtian. Even after the stay in Germany has come to an end, the Humboldt Foundation maintains close links with their alumni. The alumni sponsorship is tailored to the needs of every single Humboldtian, providing flexible support for the particular development and path in life as well as for cooperation with others. The Humboldt Network includes 49 academics who have been awarded the Nobel Prize. In 2011, the Nobel Prize for Medicine was divided among Humboldtians Bruce A. Beutler from the United States, Jules Alphonse Hoffmann from Luxembourg and Canadian Ralph M. Steinman. Beutler and Hoffman jointly received one half of the prize for their discovery of innate immunity. Steinman received the other half for his discovery of the dendritic cell and its role in adaptive immunity.From 2008 to 2012, a total of 2,830 fellowships were granted to academics abroad from all disciplines. For the same period, the Humboldt Foundation received 1,779 applications and approved 526 fellowships to researchers from the life sciences. The application and approval figures for medicine for the years 2008 to 2012 were 260 and 63, respectively.As an intermediary organisation for German foreign cultural and educational policy the Humboldt Foundation promotes international cultural dialogue and academic exchange. The Alexander von Humboldt Foundation is funded by the Federal Foreign Office, the Federal Ministry of Education and Research, the Federal Ministry for Economic Cooperation and Development, the Federal Ministry for the Environment, Nature Conservation and Nuclear Safety as well as other national and international partners.http://www.humboldt-foundation.de"} +{"text": "A grant from the Baxter Corporation was incorrectly omitted from the FundingStatement. The Funding Statement should read: \"This work is supported by grants fromthe Baxer Clinical Research Award and Renal Research Grant of Baxter Corp, China,and the ISN Research Award of ISN GO R&P Committee. The funders had no role in studydesign, data collection and analysis, decision to publish, or preparation of themanuscript.\""} +{"text": "The funders had no role in study design, data collection and analysis, decicion to publish, or preparation of the manuscript.The following funder is missing from the Funding section: The Norwegian Council for Mental Health. The complete, correct funding information is as follows: This project has been financially supported by The Norwegian Council for Mental Health and the Norwegian ExtraFoundation for Health and Rehabilitation through EXTRAfunds. Grant number: 2.13/2/0021. URL:"} +{"text": "Details of correction: addition to acknowledgmentsExisting text:The authors would like to acknowledge the guidance of biostatistician Doctor Alison Bowling with the data analysis for this project; the Australian Longitudinal Study of Women\u2019s Health for access to their data sets; the support of Professor Gita Mishra the ALSWH liaison person for our study and Doctor David Giles from the Victorian Cancer Council who constructed the DQESv2 FFQ used in this study.This work was supported by an Australian Government Research Training Programme (RTP) stipend grant for PhD candidate Megan Lee. RTP funding is an Australian support grant for domestic or international students conducting PhD or Master of Research degrees. The project received no other grants from funding agencies, commercial or not-for-profit sectors.This manuscript came from a PhD project. M. L. formulated the research question, designed the study, requested data through an EOI from the ALSWH, cleaned and analysed the data with contribution from A. B. and J. B., interpreted the findings and wrote the article under the supervision of S. S., J. Y. and J. B.The authors acknowledge no conflicts of interest for this research project.Corrected text should read:The research on which this paper is based was conducted as part of the Australian Longitudinal Study on Women\u2019s Health by the University of Queensland and the University of Newcastle. We are grateful to the Australian Government Department of Health and Aged Care for funding and to the women who provided the survey data.The authors would like to acknowledge the guidance of biostatistician Doctor Alison Bowling with the data analysis for this project; the Australian Longitudinal Study of Women\u2019s Health for access to their data sets; the support of Professor Gita Mishra the ALSWH liaison person for our study and Professor Graham Giles and Professor Roger Milne of the Cancer Epidemiology Centre of Cancer Council Victoria, for permission to use the Dietary Questionnaire for Epidemiological Studies (Version 2), Melbourne: Cancer Council Victoria, 1996This work was supported by an Australian Government Research Training Programme (RTP) stipend grant for PhD candidate Megan Lee. RTP funding is an Australian support grant for domestic or international students conducting PhD or Master of Research degrees. The project received no other grants from funding agencies, commercial or not-for-profit sectors.This manuscript came from a PhD project. M. L. formulated the research question, designed the study, requested data through an EOI from the ALSWH, cleaned and analysed the data with contribution from A. B. and J. B., interpreted the findings and wrote the article under the supervision of S. S., J. Y. and J. B.The authors acknowledge no conflicts of interest for this research project.Details of correction: changes to paper titleExisting text: Is dietary quality associated with depression? An analysis of the Australian longitudinal study of women\u2019s health dataCorrected text should read: Is dietary quality associated with depression? An analysis of the Australian Longitudinal Study on Women\u2019s Health data"} +{"text": "In the published article, there was an error in affiliation [4]. Instead of \u201cBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Engineering and Technology Research Center of Food Additives, National Soybean Processing Industry Technology Innovation Center, Beijing, China,\u201d it should be \u201cBeijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Engineering and Technology Research Center of Food Additives, National Soybean Processing Industry Technology Innovation Center, Beijing Technology and Business University, Beijing, China.\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The Data Availability Statement for this article is updathttps://doi.org/10.34894/V9IGKJ with the permission of the ethics board of the Faculty of Behavioral and Social Sciences, University of Groningen (contact: dcc@rug.nl).The data that support the findings of this study are available from DataVerseNL. Because of ethical reasons, restrictions apply to the availability of these data . Data are available at"} +{"text": "Following publication of the original article , the autAcknowledgementsThe manuscript is based on the project \u201cOslo-Rajasthan Sick Newborn Care Training Programme\u201d which was originally designed by The International Collaboration Unit, Oslo University Hospital and FK Norway, along with J K Lone Hospital, Sir Padam Pat Mother and Child Health Institute, Jaipur and Department of Medical Education and Department of Health and Family Welfare, Government of Rajasthan, India, with support from Royal Norwegian Embassy in New Delhi, India. Special thanks to the leadership of and to all health workers from SMS Medical College, India and Oslo University Hospital, Norway who made tremendous contributions to enable and develop the project. The author wishes to acknowledge sterling everyday contributions of the dedicated exchanged nurses and is particularly grateful to Dr Ramesh Choudhary for excellent collaboration and for his collection of data on exclusive breastfeeding after discharge.The Acknowledgements section has been updated in this correction and the original article has been"} +{"text": "The Funding statement is incorrect. The correct Funding statement is as follows: This project has been funded in whole or in part with MOH Indonesia and Federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, under contract Nos. HHSN261200800001E and HHSN261201500003I. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government."} +{"text": "Funding statement. The funding statement did not use the text that is on the project agreement with the funder. The funding statement was displayed as: \u201cThe research project was funded by two grants: (1) The 2019 Early Career Fellowship Program of the Organization for Women in Science for the Developing World (OWSD) and (2) The International Development Research Center (IDRC) with IDRC Grant Number: 109187-002 through the 2020 IndabaX-AI4D Innovation Grant.\u201dIn the published article, there was an error in the Funding statement appears below.The correct This work was carried out with the aid of a grant from UNESCO and the International Development Research Center, Ottawa, Canada. The views expressed herein do not necessarily represent those of UNESCO, IDRC, or its Board of Governors.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The authors regret that an incorrect grant number was shown in the Acknowledgements section of the published article. The corrected section should read:We acknowledge financial support from the General Directorate of Research Grants, \u0623\u062a-34-405, from King Abdul-Aziz City of Science and Technology (KACST), Kingdom of Saudi Arabia.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "The authors regret that the data availability statement was not correctly given in the original article. The correct statement is as shown below.10.15125/BATH-01212.Data supporting this work is freely accessible in the University of Bath research data archive at DOI: The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "In the published article, there was an error in the Funding statement.\u201cThis work was funded by the National Research Foundation (NRF) Thuthuka Programme grant 128296 to SM-M. The North-West University, South African Medical Research Council and the University of Zululand is also acknowledged. All the content expressed in this review is the official views of the authors and do not represent that of the North-West University. FM acknowledges funding by the NRF, Thuthuka grant UID 128296 linked to SM-M.\u201dThe correct Funding statement appears below.\u201cThis work was funded by the National Research Foundation (NRF) Thuthuka Programme grant 128296 to SM-M. Funding from North-West University and the University of Zululand is also acknowledged. The work reported herein was made possible through funding by the South African Medical Research Council (SAMRC) through its Division of Research of Capacity Development under the Early Investigators Programme from the South African National Treasury (funding number: HDID8682/MB2022/EIP052). The content hereof is the sole responsibility of the authors and do not necessarily represent the official views of the SAMRC. Also, all the content expressed in this review is the official views of the authors and do not represent that of the North-West University or the University of Zululand. FM acknowledges funding by the NRF, Thuthuka grant UID 128296 linked to SM-M.\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "In the published article, there was an error in the Funding statement.This study was supported by the NSCF (grant number 31870520) and China Agriculture Research System of MOF and MARA (grant number CARS-37).The correct Funding statement appears below.This study was supported by the NSCF (grant number 31872520) and China Agriculture Research System of MOF and MARA (grant number CARS-37).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Funding statement. The support of the National Science Foundation was omitted. The correct Funding statement appears below.In the original article, there was an error in the Funding\u201cThe authors thank the National Institutes of Health (R01-EB022592) and the National Science Foundation (CCF-2007807) for funding support.\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Due to a production error, the Funding statement was not provided. The correct Funding statement is as follows: This work was supported by the National Natural Science Foundation of China (81903672), Peking University People\u2019s Hospital Research and Development Funds (RS2020-04), and China International Medical Foundation (Z-2018-35-2003). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The publisher apologizes for this mistake. The original version of this article has been updated. Your article has been copyedited to ensure that we publish the highest quality work possible. Please check it carefully to make sure that it is correct and that the meaning was not lost during the process."} +{"text": "Funding statement. The funding information was not included in the final publication. The correct Funding statement appears below.In the published article, there was an error in the \u201cThis research was funded by the National Key R&D Program of China , the National Natural Science Foundation of China .\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The authors regret that the funding information in the acknowledgements of the original article was incorrect. Natural Science Foundation of China (No. 21671114 and U1804113), should be \u201cNatural Science Foundation of China (No. 21671114 and U1804131)\u201d.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "Humanities and Social Sciences Communications 10.1057/s41599-022-01097-5, published online 22 March 2022.Correction to: A change has been made to the Abstract to correct a statement about the merger of the Swedish Public Health Agency with the Institute for Infectious Disease Control.The original text read:In 2014, the Public Health Agency merged with the Institute for Infectious Disease Control; the first decision by its new head (Johan Carlson) was to dismiss and move the authority\u2019s six professors to Karolinska Institute. With this setup, the authority lacked expertise and could disregard scientific facts.The corrected text now reads:In 2014, the Public Health Agency, after 5 years of rearrangement, merged with the Institute for Infectious Disease Control, with six professors leaving between 2010 and 2012 going to the Karolinska Institute. With this setup, the authority lost scientific expertise."} +{"text": "We continue the practice of transparency of potential competing interests for us as editors of the Journal of Global Health, following the International Committee of Medical Journal Editors (ICMJE) Recommendations for the Conduct, Reporting, Editing, and Publication of Scholarly work in Medical Journals . As we eWe declare below the following conflicts of interest, in alphabetical order. We will regularly publish updated declarations when there is a change in editors\u2019 activities and relationships.Prof. Campbell is employed by the University of Edinburgh, where he holds a position as Professor in the Usher Institute, College of Medicine and Veterinary Medicine. He is also co-Director of the NIHR Global Respiratory Health Unit. He currently receives research funding from the European Commission (Innovative Medicines Initiative), WHO, UK NIHR and the Baszucki Brain Research Fund. He has received consultancy payments, paid via the University of Edinburgh, from WHO, the Bill and Melinda Gates Foundation, the UK Funding Councils (Research Excellence Framework) and Sanofi in the past 10 years.Prof. Campbell holds a position as the co-Editor in Chief. He occasionally reimburses expenses related to Journal\u2019s work, including travel and consumables. Prof. Campbell is a member of the International Society of Global Health (ISoGH) and a minority shareholder in the small publishing company JoGH LtdNumerous technical advisor appointments to WHO in the past 10 years.Prof. Rudan is employed by the University of Edinburgh, where he holds a position as Professor and Co-Director of the Centre for Global Health Research. He is also co-Director of the WHO Collaborating Centre in Population Health Research and Training.Prof. Rudan holds a position as the co-Editor in Chief. He occasionally reimburses expenses related to Journal\u2019s work, including travel and consumables. Prof. Rudan serves as the President of the International Society of Global Health (ISoGH), and is also a minority shareholder in the small publishing companies JoGH Ltd and Inishmore Laser Scientific Publishing LtdProf. Rudan has numerous technical advisor appointments to WHO, UNICEF and the World Bank."} +{"text": "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The following information is missing from the Funding statement: This study received funding from the Jiangsu Social Science Fund (18EYC006;"} +{"text": "In the published article, there was an error in the Funding statement. Previously, it read: \u201cThis work was supported by grants from the Natural Science Foundation of Zhejiang Province, Zhejiang, China (Grant No. LY20H020003) and Medicine and Health Science and Technology Plan Projects of Zhejiang Province, Zhejiang China (Grant No. 2019KY377).\u201d However, two more fundings supporting this research, the National Natural Science Foundation of China , should also be added to the Funding section. The correct Funding statement appears below.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.This work was supported by grants from the Natural Science Foundation of Zhejiang Province, Zhejiang, China (Grant No. LY20H020003), Medicine and Health Science and Technology Plan Projects of Zhejiang Province, Zhejiang China (Grant No. 2019KY377) and the National Natural Science Foundation of China .All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Correction: BMC Health Serv Res 22, 1004 (2022)https://doi.org/10.1186/s12913-022-08400-9Following publication of the original article , the autThe statement in the \u2018Funding\u2019 section originally read: This study is funded by the Norwegian Directorate of Health and the Research Council of Norway. The funding bodies had no role in the design of the study, nor in data collection, analysis, or data interpretation.The statement in the \u2018Funding\u2019 section should read: This study is funded by the Norwegian Directorate of Health and the Research Council of Norway. The funding bodies had no role in the design of the study, nor in data collection, analysis, or data interpretation.DGRdP was partially funded by the Southwest Center for Occupational and Environmental Health (SWCOEH), a National Institute for Occupational Safety and Health (NIOSH) Education and Research Center at The University of Texas Health Science Center at Houston School of Public Health, and awardee of Grant No. 5T42OH008421 from the (NIOSH)/Centers for Disease Control and Prevention.The original article has been"} +{"text": "There is an error in the Funding Disclosure. The correct Funding Disclosure reads:https://www.manchester.ac.uk/. CS and MS and the PCIE input were part-funded by the National Institute for Health Research (NIHR) Applied Research Collaboration Greater Manchester (award number: NIHR200174). MS is a NIHR Senior Investigator. CS was part-funded and RW was funded by the NIHR Greater Manchester Patient Safety Translational Research Centre (award number: PSTRC-2016-003). The views expressed are those of the authors and not necessarily those of the NIHR or the Department of Health and Social Care. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.This work was funded by an internal grant from the University of Manchester. University of Manchester website:"} +{"text": "The Administrative Data Research Network (ADRN) is a UK-wide initiative, funded by the Economic and Social Research Council (ESRC). The Network facilitates safe access to linked de-identified administrative data for research which is aimed at providing a benefit to our society. Administrative data research can provide wide ranging and longitudinal evidence for policy makers which therefore has the potential to improve our society.Recognising the importance of public confidence and trust to the success of the ADRN, the ESRC commissioned a public consultation to gauge understanding of social research and the reactions to the use of administrative data in research. A comprehensive UK-wide communications and public engagement strategy has been developed. From this a number of initiatives been introduced over the past two years to address public concerns and these have been reviewed, revised and extended as the Network has evolved.Now to extend the Network's reach, the Administrative Data Research Network is developing a UK National Citizens Panel (CP). The panel will provide a representation of public views on potential changes to Network policy, procedures, governance and service provision issues. The CP will also assist with testing our public facing communications such as events, website and promotional materials.This paper presents the previous and current public engagement initiatives that the Administrative Data Research Network has incorporated within its policies and service that enables better knowledge for a better societywww.adrn.ac.ukFunded by the Economic & Social Research Council, the ADRN, set up as part of the UK Government's Big Data initiative, is a UK-wide partnership between universities, government bodies, national statistics authorities and the wider research community."} +{"text": "The correct affiliation details for Mariana Pinto Da Costa are as follows: Mariana Pinto Da Costa, MD, MSc, PhD, Consultant Psychiatrist at South London and Maudsley NHS Foundation Trust and Senior Lecturer at the Institute of Psychiatry, Psychology & Neuroscience, King's College London, London, UKThe correct wording in the declaration of interests should be \u201cMPC is a member of the Editorial Board of The British Journal of Psychiatry and of BJPsych Advances\u201d and \u201cHE is a member of the Editorial Board of the British Journal of Psychiatry International\u201d.The authors would like to make two corrections to the above paper.The authors apologise for these errors."} +{"text": "Funding statement. The Funding statement that was published was missing a second source. The correct Funding statement appears below:In the published article, there was an error in the \u201cThe authors received funding from the Agency for Science, Technology and Research (A*STAR) Career Development Award (C210112056) and Singapore National Medical Research Council (OFYIRG19may-0007).\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "In \u201cComparing Online and On-Site Cognitive Behavior Therapy in Major Depressive Disorder: Protocol for a Noninferiority Randomized Controlled Trial\u201d :e29726), the following change was made:In the originally published article, the Conflicts of Interest section inadvertently appeared as follows:None declared.In the corrected version, the Conflicts of Interest section has been corrected as follows:NW is an employee of NexJ Health and holds stock in the company. NexJ Health provides in-kind subscriptions for the digital health platform of NexJ Connected Wellness, which enables the delivery of the CBT-M program and provides health coaching to the participants in the CBT-M intervention group. PR receives in-kind software support from NexJ Health for this investigator-initiated study, funded by the Canadian Institutes of Health Research (CIHR). He also receives research support from NexJ Health through the Digital Health Research Fund administered by the Faculty of Health at York University. ZD has received research and equipment in-kind support for an investigator-initiated study through Brainsway Inc and Magventure Inc. He is also on the scientific advisory board for Brainsway Inc. His work has been supported by the National Institutes of Mental Health (NIMH), Canadian Institutes of Health Research (CIHR), Brain Canada, and Temerty Family Foundation, and Grant Family Foundation.The correction will appear in the online version of the paper on the JMIR Publications website on April 21, 2022, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "The authors wish to make the following corrections to this paper :In the original published article, the Institutional Review Board Statement was not included. The Institutional Review Board Statement is added below.Institutional Review Board Statement: Ethics approval was obtained from each of the two institutions: The Internal Ethics Review Committee of the Koret School of Veterinary Medicine Veterinary Teaching Hospital, Israel, certificate number KSVM-VTH/07_2015 and Veterinary Research Ethics Committee, University of Liverpool, UK, certificate number VREC237.The authors apologize for any inconvenience caused and state that the scientific conclusions are unaffected. The original publication has also been updated."} +{"text": "The authors regret that the funder information in the Acknowledgements section is incorrect in the original article. The correct information is shown below.This study was supported by the Natural Science Foundation of Xinjiang, China, Grant No. 2016D01A073.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "In the published article, there was an error in the Funding statement. The funding number for Beijing Stomatological Hospital, Capital Medical University Young Scientist Program was incorrect. The corrected Funding statement appears below.\u201cThis study was supported by the National Natural Science Foundation of China (Nos. 81771094 and 82170980) and the Beijing Stomatological Hospital, Capital Medical University Young Scientist Program (No. YSP202106).\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Funding statement. The state assignment of the Ministry of Science and Higher Education of the Russian Federation was incorrectly mentioned as a funding source. The correct Funding statement appears below:In the published article, there was an error in the \u201cThis study was supported by the Russian Science Foundation, grant no. 17-15-01051.\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Capital's Funds for Research and Application of Clinical Diagnosis and Treatment Technology (No. Z191100006619121), National Center for Clinical Research in Cardiovascular Diseases (No. NCRC2020015), and Capital's Funds for Health Improvement and Research (No. 2018-2-4031).There is an error in the Funding statement. The correct order for funding information is: The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Funding statement. The published version of the Funding was, \u201cThis work is supported by the National Natural Science Foundation of China (No. 81970230 for WZ), National Natural Science Foundation of China (No. 81870292 for JW), and Major Scientific Research Project of Zhejiang Lab (No. 2019MB0AD01 for HH).\u201d There was one funding source [National Natural Science Foundation for Young Scientists of China (No. 62005248 for Z.F.)] that was omitted. The correct Funding statement appears below.In the published article, there was an error in the Funding\u201cThis work is supported by the National Natural Science Foundation of China (No. 81970230 for WZ), National Natural Science Foundation of China (No. 81870292 for J\u2019aW), Major Scientific Research Project of Zhejiang Lab (No. 2019MB0AD01 for HH), and National Natural Science Foundation for Young Scientists of China (No. 62005248 for ZF)\u201d.The authors apologize for this error and state that this does not change the scientific conclusion of the article in any way. The original article has been updated."} +{"text": "In the published article, there was an error in Funding statement. The grant number was incorrect. The correct Funding statement appears below.In the published article, there was an error in the th Nuclear Energy R&D Project (No. 20201192). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript\u201d.\u201cThis work was supported by the National Key R&D Program of China , the Natural Science Foundation of Guangdong Province , the National Natural Science Foundation of China (Nos. 82161160343 and 82002168), the Fundamental Research Funds for the Central University (No. 22qntd4813), the 111 Project (No. B12003) and the 6The authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "There is information missing from the Acknowledgments section. It should read as follows:The authors would like to thank the study participants for their time and responses without which this paper would not have been possible to write. We also thank the regional, district, shehia and village authorities where the study was conducted for granting the permission and providing every support to successfully conduct this study. We are grateful to Prof. Jennifer A. Downs of the Center for Global Health, Weill Cornell Medicine, New York, United States of America for her support in conceptualizing the project whose outputs include this paper. We also thank her for the reviews, comments and suggestions on the early drafts of this article. The authors would like to acknowledge that the article included qualitative data designed and collected in Tanzania as part of the study \u201cBringing Down Hurdles for Female Genital Schistosomiasis Access to Care: a Multi-Country Socio-Structural Integrated Approach to Developing a Community-Based Teaching Platform\u201d carried out in Zambia, Tanzania and Malawi, funded by The Task Force for Global Health NTD-SD, and led by Virginia Bond , Humphrey Mazigo and Khumbo Kalua (Blantyre Institute for Community Outreach (BICO). The views expressed in this publication are those of the authors and not necessarily those of the funding agencies. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the supporting offices."} +{"text": "In \u201cUsing Wake-Up Tasks for Morning Behavior Change: Development and Usability Study\u201d :e39497) the authors noted two errors.The affiliation of the author Jisu Ko was incorrectly mentioned as the following:Department of Human-Computer Interaction.This has been corrected to:Department of Applied Artificial Intelligence.Under \u201cAcknowledgments\u201d the original text read:This work was supported by Institute of Information & communications Technology Planning & Evaluation (IITP) grant funded by the Korea government (Ministry of Science and ICT) and Hanyang University ERICA.It has been replaced by the following:This work was supported by Institute of Information & communications Technology Planning & Evaluation (IITP) grant funded by the Korea government (Ministry of Science and ICT) ).The correction will appear in the online version of the paper on the JMIR Publications website on October 3, 2022, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "In the original article, there were some errors in the funding part and in this correction, the correct funding text is as follows:This work was supported by the National Key R&D Program of China .The Original article has been corrected."} +{"text": "Funding statement. \u201cThis work was supported by the National Natural Science Foundation of China (No. 51708326), the Major Science and Technology Program for Water Pollution Control and Treatment of China (Nos. 2017ZX07103007 and 2018ZX07111006), and the Tsinghua University Initiative Scientific Research Program (No. 2018Z02ALB01).\u201d The correct Funding statement appears below.In the published article, there was an error in the This work was supported by the State Key Joint Laboratory of Environment Simulation and Pollution Control (22Y03ESPCT).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The National Institute on Aging (NIA) at the National Institutes of Health, Department of Health and Human Services, is the federally designated lead agency on aging research and supports research to understand the nature of aging and to extend the healthy, active years of life. In the last six years, NIA has experienced a tripling of its budget. In keeping with NIA\u2019s scientific mission, funding includes significant investments in Alzheimer\u2019s disease (AD) and AD-related dementias research, as well as in non-AD research. This symposium will provide a forum for exploration of the implications of the NIA budget for the general research community. NIA\u2019s senior staff will discuss research priorities and programs supported by the Institute. A question-and-answer session will follow these remarks on current funding and future priorities and research directions of NIA."} +{"text": "In the published article, there was an error in the Funding statement. The grant number for the Science and Technology Program of Guangzhou, China was displayed as \u201c202201011577\u201d. The correct grant number is \u201c202201011571\u2019\u2019. The correct Funding statement appears below.This work was supported by the National Natural Science Foundation of China (82072196 and 81871573), the China Postdoctoral Science Foundation\u2013funded project (2020M672721), and the Science and Technology Program of Guangzhou, China (202201011571).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Following publication of the original article , the autNamely, the funding declaration has been corrected to the following:\"This work is supported by the National Research Fund of Luxembourg (Grants # C14/BM/8225223 and C17/BM/11613033 to YD), the Ministry of Higher Education and Research of Luxembourg, and the Heart Foundation - Daniel Wagner. FMS received a fellowship from the National Research Fund of Luxembourg for this work (Grant # C17/BM/11613033). This work is also supported by independent research grants from nonprofit or governmental agencies and by governmental funding of clinical research within the Swedish National Health Service.\"The authors thank you for reading and apologize for any inconvenience caused."} +{"text": "The Editor-in-Chief has retracted this article. After publication, concerns were raised regarding the ethics approval for this study. The authors have confirmed that approval from the National Ethics Committee for Health Research in Cambodia was not sought prior to the study.All authors agree to this retraction."} +{"text": "The authors regret the omission of a funding acknowledgement in the original article. This acknowledgement is given below.Ding Ding would like to acknowledge support from the Agency for Science, Technology and Research (A*STAR) AME Young Independent Research Grant, project: A1884c0020.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "In the published article, there was an error in the Funding statement. [This research was supported by a fund (code: 2019ER690200) by Research of Korea Centers for Disease Control and Prevention]. The correct Funding statement appears below.This research was supported by a fund (code: 2022-ER0507-00) by Research of Korea Disease Control and Prevention Agency.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Instead of Abcam it should have been Life Technologies, Bio-Rad and Life Technologies, respectively. The HTML and PDF versions of the Article have now been updated with the correct information."} +{"text": "In addition, the authors regret the omission of one of the authors, Stefan Krauss, from the original manuscript. The corrected author list and affiliations are as shown above.The authors regret that one of the affiliations and SP from the Norwegian Cancer Society (grant No. 190257-2017). J. W. and S. K. were supported by the Research Council of Norway , by South-Eastern Norway Regional Health Authority and from the Norwegian Cancer Society (grant No. 5803958).The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "Funding statement as published. The correct grant number for the Startup Fund for Scientific Research of Fujian Medical University is 2018QH1202. Corrected funding statement is given below:In the original article, there was a mistake in the \u201cThis work was supported by the Scientific Research Project for the Middle-aged and Youths of Fuzhou Health Department (grant number: 2019-S-wq9) and the Startup Fund for Scientific Research of Fujian Medical University (grant number: 2018QH1202).\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "An analysis of currently existing partnerships and cross-country collaboration for physical activity (PA) promotion is valuable for understanding how such partnerships operate, and how they impact national PA promotion efforts. This study aimed to outline the structure of the European Union's (EU) National Physical Activity Focal Point Network, to evaluate its outputs and benefits, and to describe its potential and challenges.We employed a mixed methods approach with three components: (1) document analysis of network meeting reports, (2) semi-structured interviews with key officials who were involved in establishing the network, and (3) an online evaluation survey with the national PA Focal Points.The PA Focal Point Network was founded in 2014, and its main task is to coordinate the collection of information for the EU's HEPA Monitoring Framework. Each of the EU Member States nominated a representative to the network. Focal Points usually meet twice a year to discuss issues related to the HEPA Monitoring Framework and to share best practices and plan activities for the promotion of PA within the EU. The results of the evaluation survey show that participation in the network helped members to specify goals for PA promotion, gain knowledge, and identify opportunities to promote PA in their country. From the perspective of the Focal Points, most helpful outputs of the Network activity are the country factsheets on physical activity, the connections within the Network and the opportunity to share their experience with colleagues during meetings and group discussions.The study shows that the PA Focal Point Network may serve as an example of successful cross-country collaboration in PA promotion. The network has supported the monitoring of the implementation of the EU Council Recommendation on HEPA across sectors in particular and of PA promotion in the EU in general. It also had positive effects on national PA promotion efforts and on cooperation between countries. All in all, the PA Focal Point Network can serve as an example for other world regions or policy areas that set up similar networks."} +{"text": "The Data Availability statement is incomplete. The complete Data Availability statement is:HAR is funded by a UNSW Scientia Program Fellowship and the Australian Research Council Discovery Early Career Researcher Award (DECRA) under grant DE220101210. HAR was also supported by a WA Department of Health Near-Miss Merit Award to HAR.ARRF is supported by an Australian National Health and Medical Research Council Fellowship APP1154524. ARRF was also supported by funds raised by the MACA Ride to Conquer Cancer and a Senior Cancer Research Fellowship from the Cancer Research Trust.HRR and RG were supported by IRN National Science Foundation (INSF), Grant No. 96006077.This work was funded by an Australian Research Council Discovery Project grant to ARRF (DP160101960). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In 2015 the German Prevention Act was implemented. The National Prevention Conference published the first National Prevention Report in 2019 to evaluate the health promotion activities. The second National Prevention Report is planned for 2023. Development of a harmonized prevention reporting system for the German Federal States is needed to form the basis for the contribution of the Federal States to the next National Prevention Report. A working group mandated from the sub-national health authorities has developed a harmonized prevention reporting system for the German Federal States since 2018. The Robert Koch Institute collaborated as representative of the national level during the process. Subject areas for indicators were selected based on a survey in which all 16 State Ministries of Health participated. Indicator subgroups developed indicators for each subject area based on predefined indicator selection criteria. Final set of indicators was adopted by indicator rating and majority voting process. The German Health Ministers Conference acknowledged the indicator system in June 2021. The conceptual framework is adapted from the health determinants model of Dahlgren and Whitehead. The indicator system is divided into 14 subject areas categorized into upstream, midstream and downstream level of prevention indicators. Seventy-three prevention indicators were included as a whole. The indicator short list consists of 32 Core indicators. An overview of the prevention indicator system will be given. First results of a pilot data collection will be shown. Health promotion and prevention reporting tools are needed to monitor prevention policies and evaluate health promotion measures. The prevention indicator system of the German Federal States will be used for the National Prevention Strategy in Germany of which one component is the next National Prevention Report 2023.\u2022\u2002The prevention indicator system of the German Federal States is a useful tool to monitor prevention policies.\u2022\u2002The indicator system will form the basis for the German Federal States\u2019 contribution to the National Prevention Report 2023."} +{"text": "In \u201cGender-Specific Impact of Self-Monitoring and Social Norm Information on Walking Behavior Among Chinese College Students Assessed Using WeChat: Longitudinal Tracking Study\u201d :e29167), one error was noted.The foundation number of the National Natural Science Foundation of China was mistaken. In the originally published paper, under \u201cAcknowledgments\u201d, the foundation information was listed as follows:This research was supported by Beijing Natural Science Foundation , the National Natural Science Foundation of China \u2026This has been corrected to:This research was supported by Beijing Natural Science Foundation , the National Natural Science Foundation of China \u2026The correction will appear in the online version of the paper on the JMIR Publications website on March 31, 2022, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "In the published article, there was an error in the Funding statement. The Funding statement was not present in the original version. The correct Funding statement appears below.\u201cThis work was financially supported by the National Natural Science Foundation of China (Award number(s): 81971194, 82171421, 91949118) and Science and Technology Commission of Shanghai Municipality (Award number(s): 2018SHZDZX01, 21S31902200); National Health Commission of the People\u2019s Republic of China.\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "A. Rogers The authors regret the omission of the following conflict of interest statement.Dr Martin J. T. Reaney is the founder of, and has an equity interest in, Prairie Tide Diversified Inc. . Dr Youn Young Shim is a Market Consultant for PTD in Korea. The terms of this arrangement have been reviewed and approved by the University of Saskatchewan in accordance with its conflict of interest policies.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "Correction: BMC Public Health 22, 475 (2022)https://doi.org/10.1186/s12889-022-12777-xThe original publication of this article containeThere was a problem concerning the variable denoting responder ID. For one panel used, all respondents were assigned unique IDs each time they completed a survey, regardless of whether they had previously completed the survey.Therefore, repeat respondents were not identified. This affected data in waves 8 to 57 (inclusive). The authors have since worked with the market research companies to rectify this problem. For another article, re-running generalised estimating equations (GEEs) using corrected responder IDs did not meaningfully change results .This should not affect the results nor conclusions of this article .Funding statementThis work was funded by the National Institute for Health Research (NIHR) Health Services and Delivery Research programme (NIHR project reference number (11/46/21)). Surveys were commissioned and funded by Department of Health and Social Care (DHSC), with the authors providing advice on the question design and selection. LS, RA and GJR are supported by the National Institute for Health Research Health Protection Research Unit (NIHR HPRU) in Emergency Preparedness and Response, a partnership between the UK Health Security Agency, King\u2019s College London and the University of East Anglia. RA is also supported by the NIHR HPRU in Behavioural Science and Evaluation, a partnership between the UK Health Security Agency and the University of Bristol. HWWP has received funding from Public Health England and NHS England. NTF is part funded by a grant from the UK Ministry of Defence. The views expressed are those of the authors and not necessarily those of the NIHR, UK Health Security Agency, the Department of Health and Social Care or the Ministry of Defence. The Department of Health and Social Care funded data collection (no grant number).Competing interests statementAll authors had financial support from NIHR for the submitted work. RA is an employee of the UK Health Security Agency; HWWP received additional salary support from Public Health England and NHS England; HWWP receives consultancy fees to his employer from Ipsos MORI and has a PhD student who works at and has fees paid by Astra Zeneca. At the time of writing GJR is acting as an expert witness in an unrelated case involving Bayer PLC, supported by LS. NTF is a participant of an independent group advising NHS Digital on the release of patient data. All authors were participants of the UK\u2019s Scientific Advisory Group for Emergencies or its subgroups."} +{"text": "Funding statement for this article was omitted:Due to a production error, the in vivo efficacy of fumarate for mitochondrial disease in vivo). Besides, we would like to acknowledge the UCLH/UCL for which PG works and receives a proportion of funding from the Department of Health's NIHR Biomedical Research Centers funding scheme, and the support given by the CRN: North Thames, National Institute for Health Research.\u201d\u201cWe would like to thank the funders that supported us, in particular FARA and an NIH grant we received (4R33NS106719-03 Pharmacodynamics and The publisher apologizes for this mistake. The original version of this article has been updated."} +{"text": "Jadhav, The authors regret the omission of the following conflict of interest statement.Dr Martin J. T. Reaney is the founder of, and has an equity interest in, Prairie Tide Diversified Inc. . Dr Youn Young Shim is a Market Consultant for PTD in Korea. The terms of this arrangement have been reviewed and approved by the University of Saskatchewan in accordance with its conflict of interest policies.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "In the published article, there was an error in the Funding statement. The funding statement for the Key Development Project of the Department of Science and Technology was displayed as \u201cPS was supported by the Grant Agency of the Czech Republic and Palack\u00fd University Grant Agency [IGA-2022_002]. JR work was supported by INTA PE-142 project. EW was supported by the USDA Hatch program through the Vermont State Agricultural Experimental Station.\u201d. The correct statement is \u201cPS was supported by the Grant Agency of the Czech Republic and Palack\u00fd University Grant Agency [IGA-2022_002]. JR work was supported by INTA PE-142 project. EW was supported by the USDA Hatch program through the Vermont State Agricultural Experimental Station. EW was also funded by the Ministry of Science and Higher Education of the Russian Federation as part of World-class Research Center program: Advanced Digital Technologies (contract No. 075-15-2022-311 dated 20.04.2022)\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "International Journal of Environmental Research and Public Health (IJERPH) has increased its publications of scientific papers related to exercise; a search of Pubmed (on 22 June 2022) using IJERPH and exercise as keywords showed 1788 entries for 2021 compared to 80 entries in 2016 [The in 2016 being th in 2016 , and ref in 2016 is to de"} +{"text": "The authors regret that incorrect details were given for ref. 52 in the original article. The correct version of ref. 52 is given below as The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "In the original article, there was an error in the Funding statement. The funder \u201cLeibniz ScienceCampus SOEP RegioHub (Bielefeld University and SOEP/DIW Berlin)\u201d was missing. The correct Funding statement appears below.This research was partly funded by (a) the Leibniz ScienceCampus SOEP RegioHub (Bielefeld University and SOEP/DIW Berlin) and (b) the German Ministry for Family Affairs, Senior Citizens, Women and Youth (BMFSFJ) in the context of the National Discrimination and Racism Monitor (NaDiRa) as well as the Research Association Discrimination and Racism (FoDiRa) of the DeZIM Research Community (German Center for Integration and Migration Research).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "After this article was publThis study involved human stool samples that originated in Nepal. The samples were collected at Tribhuvan University Teaching Hospital, Microbiology and Public Health Research Laboratory, Kathmandu, Nepal; transferred to the University of Georgia in 2012; and provided to the US Food and Drug Administration (FDA) in 2015. As of the time of this notice, PLOS has been unable to reach the National Health Research Council (NHRC) in Nepal to clarify if the study complied with Nepalese regulations for human subjects research and/or transfer of human fecal samples from Nepal to the University of Georgia and then to the FDA.Cyclospora cayetanensis oocysts was approved by the FDA Research Involving Human Subjects Committee (RIHSC) in 2010\u20132013 (#10-095F). However, neither University of Georgia nor Tribhuvan Teaching Hospital were specified as potential sources for specimens in the RIHSC approval document; instead, another US-based institution was named. This discrepancy has not yet been resolved. PLOS has contacted the FDA.A protocol for collection and analysis of anonymized fecal specimens for the isolation of The University of Georgia Office of Research evaluated the study\u2019s compliance with applicable policies, regulations, and guidance, and noted that University of Georgia IRB concluded the study neither qualified as human subjects research nor required further IRB review or approval. The project received approval of the University of Georgia Institutional Biosafety Committee in 2010 and 2015. The University of Georgia IRB and Biosafety Committee reviewed research conducted with these samples at the University of Georgia. The work conducted at the FDA and the use of the samples at the FDA were not assessed by the University of Georgia committees.PLOS ONE Editors issue this Expression of Concern to notify readers of the unresolved issues discussed above.The"} +{"text": "Ref. 28 in the published article was incorrect, with an incorrect page range provided. The correct version is shown as The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "The following information is missing from the Funding statement: This work was supported by the UCLA AIDS Institute, the James B. Pendleton Charitable Trust, and the McCarthy Family Foundation."} +{"text": "Natalia Polouliakh. As well as having affiliation(s) Department of Ophthalmology and Visual Sciences, Yokohama City University Graduate School of Medicine, Yokohama, Japan and Sony Computer Science Laboratories Inc., Tokyo, Japan, Natalia Polouliakh should also have Scientista Co., Ltd., Tokyo, Japan.In the published article, there was an error regarding the affiliations for In the published article, the Conflict of Interest for Natalia Polouliakh (NP) was not sufficiently disclosed. The Conflict of Interest should have contained the following disclosure:NP is an employee of Sony Computer Science Laboratories, Inc. and also the president and CEO of Scientista Co., Ltd. These companies did not provide funding for this study. Scientista Co., Ltd. sells a cosmetic compounded with alpha-arbutin: however, this situation did not affect the results reported in this study.The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "In the original article, we neglected to include the funder Zhejiang Province Public Welfare Technology Application Research Project, Grant No. LGF21H090004 to Yuchen Ying, the funder Medical and Health Science and Technology Plan Project of Zhejiang Province, Grant No. 2021KY335 to Yuchen Ying, and the funder Science and Technology Innovation Activity Plan of Zhejiang University Student & XinMiao Talents Program, Grant No. 2021R464003 to Yuchen Ying. The updated funding statement appears below:This work was supported by the National Key R&D Program of China [Grant Nos. 2017YFC1310902 and 2018YFC1315305]; Ningbo Health Branding Subject Fund [Grant Nos. PPXK2018-01 and PPXK2018-02]; Sanming Project of Medicine in Shenzhen [Grant No. SZSM201803080]; The Special Innovation Program of Guangdong Education Department [Grant No. 2018GKTSCX077]; and Natural Science Foundation of Zhejiang Province [Grant No. LQ20H020001], Zhejiang Province Public Welfare Technology Application Research Project [Grant No. LGF21H090004] to YY, Medical and Health Science and Technology Plan Project of Zhejiang Province [Grant No. 2021KY335] to YY, and Science and Technology Innovation Activity Plan of Zhejiang University Student & XinMiao Talents Program [Grant No. 2021R464003] to YY. The funding body had no further role in the study design, the collection, analysis, and interpretation of data, the writing of the manuscript and the decision to submit the paper for publication.In the published article, there was also an error regarding the affiliation(s) for Yuchen Ying. As well as having affiliation 1, they should also have Department of Elderly Health Care and Management, School of Health Services and Management, Ningbo College of Health Sciences, Ningbo, China.The authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The attention of the Chinese government on nutrition, exercise, and health refers to the attention degree of the Central Government to the nutrition, exercise, and health of Chinese nationals and reflects whether Central Government attach importance to Chinese nationals' nutrition, exercise and health or not and the distribution of resources, which influence the physical quality and health level of Chinese nationals. Based on the attention theory and attention distribution proposed by Herbert Simon, Dai Kai, et al., this study took 43 Central Government Work Reports from 1978 to 2020 as research samples, used literature reviews, and textual analysis methods, and applied the Nvivo12.0 software to conduct qualitative and quantitative analyses about the contents of the Central Government Work Report concerning the nutrition, exercise, and health of Chinese nationals. This research found the following: (1) There has been a relatively huge overall change in the attention of the Central Government, that is, the level of attention, to the nutrition, exercise, and health of Chinese nationals from 1978 to 2020, and the policies related to nutrition, exercise and health of Chinese nationals issued by the Central Government have been growing faster. (2) The income level of the urban and rural residents, the total production of various types of food, dietary structure, the total number of medical and health institutions, the average life expectancy of the Chinese population, and the number of sports venues have been constantly increasing since the reform and opening up, which has effectively promoted the improvement of the nutrition, exercise and health level of Chinese nationals, and it cannot be achieved without the attention and support of the Central Government. However, the change in the lifestyle of Chinese nationals has led to the growth of the modern \u201cCivilization Disease,\u201d which is also an important issue that the Central Government needs to handle urgently. The Outline and Plan of \u201cHealthy China 2030\u201d issued by the Communist Party of China of Central Committee and the State Council of the People's Republic of China (hereinafter referred to as State Council) that is the Central Government of the People's Republic of China proposed that \u201chealth is an inevitable requirement for promoting human beings' all-round development, and the realization of national health and longevity is an important symbol of the country's prosperity, strength, and national rejuvenation\u201d . Does thKey Points of Works\u201d that was issued by the Ministry of Education of the People's Republic of China from 1987 to 2019 through five dimensions, and uncovered the characteristics of the attention distribution of the Central Government to vocational education, and put forward suggestions to optimize the attention distribution levels of the Central Government to vocational education \u201d and \u201cRegulations on the Work of Rural National Communes \u201d, which promoted the development of the production undertakings in China, such as agriculture, animal husbandry, forestry, sideline, and fishery. In 1982, the attention level of the Central Government to the nutrition, exercise, and health of Chinese nationals reached 1.6% that because, in January 1982, the CPC Central Committee approved the \u201cMinutes of the National Rural Work Conference No.1)\u201d, and pointed out that \u201cthe production team should make plans for the comprehensive development of agriculture, forestry, animal husbandry, sideline, fishery, industry, and commerce in accordance with local conditions, and improve the circulation of rural commodities\u201d. In addition, the Central Patriotic Health Campaign Committee and the Ministry of Health of the People's Republic of China jointly issued the \u201cNotice on Further Development of Patriotic Health Campaigns and the Construction of Socialist Spiritual Civilization.\u201d The attention level of the Central Government to the nutrition, exercise, and health of Chinese nationals was as low as.56% in 1984. The reason for this phenomenon was that the CPC Central Committee issued the \u201cSome Issues of Current Rural Economic Policy No.1)\u201d and \u201cNotice on Rural Work in 1984 No.1)\u201d which have effectively promoted the all-round development of agriculture, forestry, animal husbandry, sideline and fishery in China, and the living standards of the urban and rural residents were further improved. As a result, the number of the literal expressions on nutrition in the Government Work Reports have declined in recent years, and the Central Government has laid its main attention on the system reform and opening up. Compared with 1984, the attention level of the Central Government to the nutrition, exercise, and health of Chinese nationals increased from 1985 to 1991 and reached a peak state of 1.61% in 1988. In October 1984, the CPC Central Committee issued the \u201cNotice on Further Development of Sports,\u201d which once again stressed that \u201csports is closely related to Chinese nationals' health, and we must adhere to the policy of combining popularization and improvement, focusing on school sports\u201d. In January 1985, the CPC Central Committee and State Council issued \u201cTen Policies on Further Activating the Rural Economy No.1),\u201d which was beneficial in improving the tight supply of agricultural products and promoting the rationalization of the rural industrial structure and the diversification of the diet of the urban and rural residents. In April of the same year, the State Council approved the \u201cReport on Several Issues of Health Work Reform (Released by State Council (1985) NO.62)\u201d by the Ministry of Health of the People's Republic of China, which unveiled the prolog of health reform and promoted the development of Medical and Health Care in China. In January 1987, the CPC Central Committee issued a notice on \u201cleading Rural Reform deeper No.1\u201d) which was an important reason for the peak of the attention of the Central Government in 1998. In January 1989, the State Council approved and forwarded the \u201cOpinions on Issues Related to the Expansion of Medical and Health Services\u201d issued by the Ministry of Health, Ministry of Finance, Ministry of Personnel, State Price Bureau, and State Administration of Taxation of the People's Republic of China. However, the attention level of the Central Government to the nutrition, exercise, and health of Chinese nationals fell to 0.51% in 1992, for the reason that: In the 1990s, the State Council approved the \u201cRegulations on School Sports Work and Regulations on School Health Work\u201d that was beneficial to improving the physical health level of the youth, so as to enhance the physical quality and health level of Chinese nationals; In October 1991, the State Council issued a \u201cNotice on Further Invigorating the Circulation of Agricultural Products\u201d, which was beneficial to solving the problem of the lag in the circulation of agricultural products and laying the foundation for the urban and rural residents to purchase diversified food, obtain the nutrients that their bodies need, and promote good health. Hence, the level of attention of the Central Government was low in 1992.Bryan Jones believed that \u201call kinds of factors in the decision-making situation have an effect on decision-making\u201d . In diffOpinions on Deepening the Reform of the Health and Medical System\u201d in 1992, which proposed to reform the health management system and the price system of Medical and Health Services. In addition, according to the general goal of establishing a socialist market economy system proposed by the 14th National Congress of the Communist Party of China, the State Council issued the \u201cNotice on Accelerating the Reform of Grain Circulation System\u201d to promote the reform of the grain circulation system toward commercialization and market-oriented operation in February 1993. In 1995, the attention of the Central Government to the nutrition, exercise, and health of Chinese nationals increased to 1.18%. The main reasons are as follows: In May 1993, State Council issued the \u201cOutline for the Reform and Development of China's Food Structure in the 1990s (Released by State Council (1993) NO.40),\u201d which was the first document on food nutrition that was issued by the country since the founding of the People's Republic of China; the State Council issued the \u201cRegulations on the Administration of Medical Institutions (Released by State Council (1994) No.194)\u201d; the CPC Central Committee and State Council issued the \u201cOpinions on Agricultural and Rural Work in 1994 No.4)\u201d and \u201cOpinions on Doing the Work in Agriculture and Rural Work well in 1995 No.6).\u201d From 1996 to 1999, the attention of the Central Government to the nutrition, exercise, and health of Chinese nationals varied little and the trend remained relatively stable. The following reasons could explain this phenomenon: Firstly, in June 1995, the \u201cOutline for the National Fitness Program (Released by State Council (1995) No.14)\u201d proposed that \u201cby 2010, the Chinese national physique and health level will be comprehensively improved, and National Fitness system with Chinese characteristics will be basically built\u201d, and in August of the same year, the 15th plenary session of the Standing Committee of the 8th National People's Congress adopted the \u201cSports Law of the People's Republic of China\u201d, which clearly put forward the requirement that \u201csports work should adhere to the development of National Fitness Activities as the basis for the development of mass sports activities to improve the physical fitness of the whole nation\u201d; secondly, in January 1997, the CPC Central Committee and State Council promulgated the \u201cDecision on Health Reform and Development No.3)\u201d, which explicitly proposed the goals and guidelines of health work and the principles that need to be followed; thirdly, in October 1998, the \u201cResolution on Several Major Issues in Agriculture and Rural Work\u201d adopted by the Third Plenary Session of the 15th CPC Central Committee proposed that \u201cwe must adhere to market-oriented reforms, steadily develop food production, combine agriculture, forestry, animal husbandry and fishery, and ensure the effective supply of agricultural products\u201d. From 2000 to 2002, the reasons for the low attention level of the Central Government to the nutrition, exercise, and health of Chinese nationals were: the National Sports Work Conference held in Beijing in 1999, which discussed the \u201cOutline of Sports Reform and Development from 2001 to 2010\u201d; In December 2000, the \u201cOutline of Sports Reform and Development from 2001 to 2010\u201d mentioned that \u201cthe main goals of sports reform and development including an obvious increase in the popularity of mass sports, the full realization of the National Fitness Plan, and the effective enhancement of Chinese nationals' physical quality\u201d; In November 2001, the General Office of State Council issued the \u201cOutline for the Development of Food and Nutrition in China from 2001 to 2010 No.86),\u201d which pointed out that \u201centering the new century, accelerating food development, improving food structure, raising the nutritional level of the whole people and improving people's physical health are the urgent needs for the improvement of the national overall quality\u201d; In January 2002, the CPC Central Committee and State Council issued the \u201cOpinions on Doing the Agricultural and Rural Work Well in 2002 No.2)\u201d, which put forward that \u201cby 2010, the rural health service system and the rural cooperative medical system adapted to the requirements of the socialist market economy system will be basically established in national rural areas\u201d. From 2003 to 2012, the attention of the Central Government to the nutrition, exercise, and health of Chinese nationals was high and peaked in 2008 at 2.21%. In July 2002, the CPC Central Committee and State Council issued the Opinions on Further Strengthening and Improving Sports Work in the New Era which stated that \u201ccarrying out National Fitness Activities and enhancing nationals' physical quality is the fundamental task of sports work\u201d. In October of the same year, the CPC Central Committee and State Council issued the \u201cDecision on Further Strengthening Rural Health No.13).\u201d As the key point of Chinese health work, rural health work was linked to the development of the rural productivity and influenced the overall physical fitness and health level of Chinese nationals. In 2003, the severe acute respiratory syndrome (SARS) epidemic situation strengthened the construction of the national public health system and increased the investment of medical and health resources. In January 2005, the CPC Central Committee and State Council issued the \u201cOpinions on Several Policies to Further Strengthen Rural Work and Improve Comprehensive Agricultural Production Capacity No.1\u201d, which put forward new requirements for Chinese agricultural and rural work. The reasons for the peak level of the attention of the Central Government to the nutrition, exercise, and health of Chinese nationals in 2008 were that: the CPC Central Committee and State Council promulgated the \u201cOpinions on Developing Modern Agriculture and Solidly Promoting the Construction of a New Rural No.1)\u201d and \u201cOpinions on Effectively Strengthening Agricultural Infrastructure and Further Promoting Agricultural Development and Increasing Farmers' Income No.1),\u201d both of which emphasized the issues about agriculture, rural, and farmers and the agricultural product safety question; the CPC Central Committee and State Council issued the \u201cOpinions on Strengthening Youth Sports and Enhancing Youth Physical Fitness No.7),\u201d which promoted the healthy growth of the youth and played a role in pushing the development of national health. In March 2009, the \u201cOpinions on Deepening the Reform of Medical and Health Care System No.6)\u201d was issued, which promoted the development of medical and health care and ensured the health of the urban and rural residents in China. In August of the same year, the release of the \u201cNational Fitness Regulations\u201d not only promoted the development of national fitness activities but also improved the physical quality and health level of Chinese nationals. In January 2010, the CPC Central Committee and State Council issued the \u201cSeveral Opinions on Increasing Efforts to Coordinate Urban and Rural Development and Further Strengthening the Foundation of Agricultural Rural Development No.1),\u201d which put forward \u201caccelerating the construction of the quality and safety supervision system and inspection and detection system for agricultural products, and actively developing pollution-free agricultural products, green food, organic agricultural products.\u201d Therefore, the Central Government has paid high attention to the nutrition, exercise, and health of Chinese nationals since 2003.The convening of the 14th National Congress of the Communist Party of China marked that the reform and opening up of China has entered a new stage, and the goal of establishing a socialist market economic system has been made. In this context, the attention of the Central Government to the nutrition, exercise, and health of Chinese nationals had also changed. Overall, the attention intensity of the Central Government to the nutrition, exercise, and health of Chinese nationals has changed greatly during this period. In 1993, the attention of the Central Government to the nutrition, exercise, and health of Chinese nationals was as low as.56%. The main reason was that the State Council issued the \u201cOutline for the Development of Food and Nutrition in China from 2014 to 2020 No.3),\u201d proposing that \u201cChina's food production is not able to meet the nutritional needs yet, the residents are undernourished and surplus coexist, and the lack of nutritional and health knowledge, which must be given great attention\u201d. In October 2016, the CPC Central Committee and State Council issued the \u201cOutline and Plan of \u2018Healthy China 2030'.\u201d The outline proposed that \u201cwe should formulate and implement a national nutrition plan, deeply carry out the research on the evaluation of the nutritional functions of food, comprehensively popularize dietary nutritional knowledge, issue dietary guidelines suitable for the characteristics of different groups of people, guide residents to form scientific dietary habits, and promote the construction of a healthy diet culture\u201d. In July 2017, the General Office of the State Council issued the \u201cNational Nutrition Plan from 2017 to 2030 No.60),\u201d which put forward that \u201cnutrition is an important material basis for human beings to maintain life, growth, and health, and nationals' nutrition is related to the improvement of nationals' quality and economic and social development.\u201d Since the 18th National Congress of the Communist Party of China, the Central Government has also made a series of new deployment arrangements for the work of national fitness. In October 2014, the State Council issued \u201cSeveral Opinions on Accelerating the Development of the Sports Industry And Promoting Sports Consumption No.46\u201d) that put forward \u201cpromoting National Fitness as the national strategy of China\u201d, which marked a significant leap in the concept and practice of mass sports development in China, recognizing national fitness as the rightful meaning of national health. National fitness is a significant means of national health, it is inevitable to promote the development of national health. In June 2016, the State Council issued the \u201cNational Fitness Plan from 2016 to 2020 No.37).\u201d In October of the same year, the \u201cOutline and Plan of \u2018Healthy China 2030\u201d' was issued by the CPC Central Committee and State Council which was a program of action to promote a healthy China, and \u201cCo-construction and sharing, national health\u201d was the strategic theme of building a healthy China. To accelerate the construction of sports power and to vigorously promote the in-depth integration of national fitness and national health, the General Office of the State Council issued the \u201cNotice on Printing and Distributing the Outline of Building Sports Power No.40\u201d in September 2019. In September of the same year, for the sake of actively implementing the national fitness initiative and making regular participation in physical exercise a way of life, the General Office of the State Council issued the \u201cOpinions on Promoting National Fitness and Sports Consumption and Pushing the High-Quality Development of the Sports Industry No.43).\u201d In October 2020, the General Office of the State Council issued the \u201cOpinions on Strengthening the Construction of National Fitness Facilities and Develop Mass Sports No.36)\u201d to boost the construction of fitness facilities, promote the vigorous development of mass sports, and enhance the level of public services for national fitness. The Central Government has also carried out a comprehensive reform in the development of the medical and health services in China. In September 2013, the State Council issued \u201cSeveral Opinions on Promoting the Development of Health Service Industry No.40),\u201d which proposed that \u201caccelerating the development of the health service industry is an inevitable requirement for deepening medical reform, improving Chinese nationals' livelihood and enhancing the health quality of the whole Chinese.\u201d In March 2015, the General Office of the State Council issued the \u201cNotice on Printing and Distributing the Outline of the National Medical and Health Service System Planning from 2015 to 2020 No.14)\u201d to promote the further optimal allocation of the Chinese medical and health resources and build an integrated medical and health service system. In August 2018, the General Office of the State Council issued the \u201cNotice on Printing and Distributing the Reform Plan for the Division of Financial Affairs Powers and Expenditure Responsibilities between the Central and Local Governments in the Field of Medical and Health No.67).\u201d In June 2019, the General Office of the State Council issued the \u201cNotice on Printing and Distributing the Key Work Tasks for Deepening the Reform of the Medical and Health System in 2019 No.28),\u201d proposing that \u201cputting Chinese national health at the center, implementing prevention as the main work, strengthening disease prevention and health promotion, deepening the linkage reform of medical care, medical insurance and medicine, and firmly promoting the reform of the Medical and Health System effectively implemented and benefiting the Mass\u201d. In July 2020, the General Office of the State Council issued the \u201cKey Work Tasks for Deepening the Reform of the Medical and Health System in the Second Half of 2020 No.25),\u201d which put forward \u201cstrengthening the construction of the public health system and further implement the Healthy China Initiative\u201d.The convening of the 18th National Congress of the Communist Party of China marked a new era of socialism with Chinese characteristics . The attFrom the above, it can be seen that since the new era, although the Central Government has paid less attention to the nutrition, exercise, and health of Chinese nationals compared with the previous decade, there has been an increasing number of policies at the central level related to nutrition, exercise, and health after entering the new era, most of which were macro-policies and planning programs. On one hand, the promulgation of these policies showed that the CPC Central Committee and the State Council have attached great importance to the nutrition, exercise, and health of Chinese nationals, and to a certain extent promoted the improvement of national nutrition, exercise, and health in China. On the other hand, it indicated the lack of micro and detailed measures on nutrition, exercise, and health. The nutrition policy and action plan of China have promoted the overall nutritional status of Chinese residents to some extent, but there are still some deficiencies that need to be improved. For example, the lack of laws and regulations on nutrition policy protection, lack of long-term mechanism and sustainability, inadequate nutrition policy system, and so on . In addiSince the reform and opening up, the attention of the Central Government to the nutrition, exercise, and health of Chinese nationals has changed significantly. The number of policies related to nutrition, exercise, and health issued by the Central Government has increased, and the level of nutrition, exercise, and health of Chinese nationals has also changed constantly. The per capita income of the Chinese urban and rural residents has been increasing since the reform and opening up. As shown in Outline for the Reform and Development of China's Food Structure in the 1990s (Released by State Council (1993) NO.40).\u201d From the early 1990s to the beginning of 2000s, the average daily intake of other cereals, fruits, and salt declined slightly, while the average daily intake of tubers, flour and its products, dark-colored vegetables, and light-colored vegetables decreased sharply. The average daily intake of rice and its products, milk and its products, livestock and poultry meats, and vegetable oils increased significantly, while the average daily intake of legumes, eggs and its products, fish, and shrimp, and animal oils increased slightly. In addition, the average daily intake of Chinese residents when it comes to energy, protein, carbohydrates, and dietary fiber declined, while the average daily intake of fat rose. After entering the 21st century, to promote the development of the Chinese national dietary structure into a nutritious and scientific way, the General Office of the State Council issued the \u201cNotice on Printing and Distributing the Outline for Chinese Food and Nutrition Development from 2001 to 2010 No.86).\u201d From 2002 to 2012, the average daily intake of other cereals, tubers, rice and its products, and legumes decreased. Meanwhile, the average daily intake of dark-colored vegetables, light-colored vegetables, fruits, fish and shrimp, milk and its products, animal oils, and salt decreased slightly. On the contrary, the average daily intake of flour and its products, eggs and its products, livestock and poultry meats, and vegetable oils all increased. Moreover, the average daily intake of energy, protein, fat, carbohydrates, and dietary fibers of Chinese residents decreased. To address the coexistence of under-nutrition and over-nutrition and to optimize the dietary structure of Chinese residents, in January 2014, the General Office of the State Council issued the \u201cOutline for the Development of Food and Nutrition in China (2014\u20132020) No.3).\u201dChina respectively conducted five national nutrition surveys in 1959, 1982, 1992, 2002, and 2012, and the researchers analyzed the changes in the dietary structure of Chinese residents by using the data of national nutrition surveys comprehensively since the reform and opening up. As seen in Notice on Printing and Distributing China's Mid-and Long-term Plan for the Prevention and Treatment of Chronic Diseases (Released by State Council Office (2017) No.12)\u201d in February 2017, which mentioned, \u201cstrengthening health education and improving the health quality of the whole people, implementing early diagnosis and treatment to reduce the risk of disease among high-risk groups, promoting the coordination of medical treatment and prevention, realizing the whole process of health management and other measures\u201d (Notice on the adjustment plan of temporary import tariff rate in 2020 (Released by State Council Customs Tariff Commission (2019) No.50),\u201d which put stated \u201cimplementing zero tariff on several new diabetes drugs.\u201d No.6)\u201d issued by the Central Government and State Council in March 2009, which has not only promoted the development of medical and health undertakings in China but also laid a foundation for the improvement of the whole national health. In addition, the average life expectancy of the Chinese population has increased from 68 years old in 1978 to 77.3 in 2019. The thought of \u201cexercise is a valuable medicine\u201d has existed since ancient times. The leaders of the past generations of the Chinese nation have attached great importance to the physical quality and health level of Chinese nationals and emphasized the importance of exercise and fitness. In June 1952, Chairman Mao Zedong wrote an inscription for the foundation of the All-China Sports Federation which stated to \u201cdevelop sports and build up the people's physical fitness.\u201d In August 1997, Chairman Jiang Zemin wrote an inscription for the national fitness work which stated that \u201cnational Fitness benefits the country and Chinese nationals and contributes to the present and future generations\u201d. In October 2005, when General Secretary Hu Jintao attended the 10th National Games in Nanjing, he proposed that \u201ccarrying out national physical fitness activities to improve the health quality of the whole nation.\u201d In August 2013, when meeting with the national mass sports advanced units and advanced individual representatives, General Secretary Xi Jinping emphasized that \u201cnational fitness is the foundation and guarantee for all people to enhance their physical fitness and lead a healthy life, national fitness is an important connotation of building moderately prosperous society in all respects, and it's also a vital basis for every person to grow up and live a happy life\u201d. It can be seen from Besides the income of urban and rural residents, dietary structure, and lifestyle, many other factors are affecting the health of Chinese nationals, such as health care, sports, and fitness. Medical and health care are related to the health of hundreds of millions of people. Since the reform and opening up, the development level of medical and health care in China has gradually increased, and the average life expectancy of the Chinese population has also risen. As can be seen in In conclusion, there has been a relatively huge overall change in the attention of the Central Government, that is, the level of attention, to the nutrition, exercise, and health of Chinese nationals from 1978 to 2020, and the policies related to the nutrition, exercise, and health of Chinese nationals issued by the Central Government have been growing faster. The income level of the urban and rural residents, the total production of various types of food, dietary structure, the total number of medical and health institutions, the average life expectancy of the Chinese population, and the number of sports venues have been constantly increasing since the reform and opening up, which has effectively promoted the improvement of the nutrition, exercise, and health level of Chinese nationals, and it cannot be achieved without the attention and support of the Central Government. However, the change of the lifestyle of Chinese nationals has led to the growth of the modern \u201cCivilization Disease,\u201d which is also an important issue that the Central Government needs to handle urgently. Therefore, the research can not only offer a window into the changes of the attention of the Central Government to the nutrition, exercise, and health of Chinese nationals since the reform and opening up, but also verify the hypothesis that the attention of the government influences the level of the nutrition, exercise, and health of Chinese nationals. Furthermore, the research also summarizes the progress and defects of the development in nutrition, exercise, and health in China to lay a good foundation for the promotion of a healthy China.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.TZ and JL: data analysis and review and editing. JL: methodology. YZ: check the data. WZ and YZ: draft the paper. All authors have read and agreed to the published version of the manuscript.This research was funded by the National Social Science Foundation of China (Grant Number: 17BTY077) and the Chinese Postdoctoral Science Foundation (Grant Number: 2016M591116).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The funders had no role in the study design, analysis, decision to publish, or preparation of the manuscript.The following information is missing from the Funding statement: This work was supported by Trond Mohn foundation, grant number TMS2019TMT02. The hosting for the computations were performed on the Norwegian Research and Education Cloud (NREC), using resources provided by the University of Bergen and the University of Oslo ("} +{"text": "The authors wish to make the following corrections to this paper . In the Dr. SangJoon Lee has been requested by Ulsan National Institutes of Science and Technology (UNIST) that his affiliation of UNIST be removed from the Affiliations, because Dr. Lee\u2019s position was not officially assigned by UNIST when the paper was published.2: Department of Infection Biology, Faculty of Medicine, University of Tsukuba, Ibaraki 305-8577, Japan. The corrected affiliation section should be: SangJoon LeeThe authors apologize for any inconvenience caused and state that the scientific conclusions are unaffected. The original publication has also been updated."} +{"text": "Following publication of the original article , the aut31770104, 31970065, U1902214).This work was supported by grants from the National Natural Science Foundation of China ("} +{"text": "In the original article, we neglected to include the funder Cancer Prevention & Research Institute of Texas, RP200023, to JI.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Following publication of the original article , the autInstituto de Salud Carlos III through the project RD16/0001/0010 . Funding has allowed the evaluation work package of the project to design the study, collect and analyse data and in writing the manuscript. AM was supported by the UK Medical Research Council (grant number MC_UU_12017/14) and the Scottish Government Chief Scientist Office (grant number SPHSU14).This project was funded by research grants of the European Commission under the 7th Framework Programme and of the The original article has been"} +{"text": "There is an error in the Funding statement. The correct number for Foundation of National Facility for Translational Medicine (Shanghai) is TMSK-2020-116.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The National Academy for State Health Policy hosts both the RAISE Act Family Caregiving Resource and Dissemination Center and the Hub for State Strategies to Build and Support Palliative Care, with generous funding from The John A. Hartford Foundation. The value of supporting individuals with serious illness and complex conditions as well as their family caregivers through telehealth, care management, advance care planning, and other added family caregiver supports has been especially evident during the COVID-19 pandemic. Policymakers are now grappling with how to restructure hard-hit health care and long-term services and supports systems to better support these individuals and their family caregivers. The State Hub provides concrete resources for states working to implement and expand high-quality palliative care, and the RAISE Center is assisting the Family Caregiving Advisory Council with creating the country\u2019s first national Family Caregiver Strategy."} +{"text": "The authors regret that the original version of this paper This study was supported by funding from National Natural Science Foundation of China and National Key Research and Development Program in China (No. 2016YFB1101100 to Dr. Yubo Fan). Other funds were obtained from Grant-in-Aid from the Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development, Biomedical Laboratory Research and Development, Washington, DC (No. I01BX000282 to Jawahar L Mehta). Additional support was provided by Stebbins Chair in Cardiology funds to Jawahar L Mehta."} +{"text": "There is an error in affiliation 5 for authors Melissa Selb and Michaela Coenen. The correct affiliation 5 is: ICF Research Branch, Nottwil, Switzerland.The ICF Research Branch is no longer a cooperation partner of the WHO Collaborating Centre for the Family of International Classifications (at DIMDI)."} +{"text": "There is an error in the affiliations for author BR Ansil. Manipal Academy of Higher Education, Manipal, Karnataka, India, should not be listed. The correct affiliation for BR Ansil is as follows: National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India.Additional information is provided regarding study methodology. The anonymity of human participants was maintained throughout the study. No live animals were handled or euthanized for the purpose of this study and only dead bats collected from villagers following an annual community event were used.A land use permit from Government of Nagaland, Office of the Additional Deputy Commissioner, Pungro, Nagaland, India;Security and sensitivity clearance of the competent authority, Department of Atomic Energy, the parent funding body of National Centre for Biological Sciences, Tata Institute of Fundamental Research (NCBS-TIFR);Permission from the Bomrr Clan for research at the privately owned study site;Approval for an exemption from the National University of Singapore (NUS) Institutional Review Board;Approval from the NUS Institutional Animal Care and Use Committee;Approval from the National Centre for Biological Sciences (NCBS) Institutional Ethics Committee (Human Subjects);Approval from the NCBS Institutional Animal Ethics Committee;Approval from the NCBS Institutional Biosafety Committee.The authors confirm that the study received all necessary approvals from the necessary national statutory bodies. The following permits and approvals were obtained:Copies of documentation for the above permits and approvals have been provided to the journal editors.At the time of publication of this notice, PLOS has been unable to reach the Indian Council of Medical Research to discuss the government-level regulations and permits relevant to this study. However, the Director of NCBS-TIFR confirmed to the journal editors that this study complied with all local regulations and received all necessary permissions, permits and had necessary approval of the parent Department i.e. the Department of Atomic Energy, Government of India."} +{"text": "In the original article, we neglected to include the funding information. The corrected Funding statement is shown below:This study was supported by the Department of Education in Guangdong Province of China, Grant No. 2016WQNCX130; Educational Science Research Institute of Shenzhen, Grant No. zdfz18007; and Shenzhen Social Science Association, Grant No. SZ2019D055 to XL.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "In this profile, Kristine M. Alpi, AHIP, FMLA, Medical Library Association (MLA) president, 2021\u20132022, is described as committed to public health, professional development, and the growth and evolution of MLA. She teaches and speaks on the shared health impact from interactions among animals, humans, and the environment, and she mentors graduate students and fellows in librarianship and informatics. Alpi earned her PhD in educational research and policy analysis in 2018 and directs the Oregon Health & Science University Library. It's a distinct honor to be able to tell you about the career of Kristine Markovich Alpi, Medical Library Association (MLA) president for 2021\u20132022.I first met Kris when she arrived at the New York Academy of Medicine, where she was starting a job as education coordinator for what was then the Region 1 Regional Medical Library. She had, however, already begun preparing herself for excellence in library services, having worked as a hospital librarian in Indiana and then participating in the National Library of Medicine (NLM) Associate Fellowship Program.Once settled in New York, Kris pursued her master's in public health, enrolling in the Hunter College School of the Health Professions. After working as an information services librarian and lecturer at the Weill Cornell Medical College, she took on the position of library manager at the New York City Department of Health & Mental Hygiene's Public Health Library, where she directly served the public health professionals that served the largest city in the United States. She also continued as a lecturer in public health at Weill Cornell, teaching students in evidence-based medicine, epidemiology, and biostatistics.With her relocation to North Carolina as director of the William R. Kenan, Jr. Library of Veterinary Medicine at North Carolina State University (NCSU), Kris entered a new area of public health\u2014that of the shared health impact from interactions among animals, humans, and the environment. Her recent coauthored article that appeared in the NLM's Director's Blog outlines the importance of One Health\u2014these shared public health impacts . She conDecember 2018 began a new phase in Kris's career as she moved to Portland and assumed the directorship of the Oregon Health & Science University Library. As part of her responsibilities as university librarian and associate professor in the Department of Medical Informatics & Clinical Epidemiology, she still educates students on informatics and epidemiology and serves as a mentor to graduate students and fellows.Kris's work in public health has extended to educating consumers by locating accurate and timely web-based information. From 1998 to 2009, she used her expertise in Spanish to build the Spanish side of the bilingual web portal NOAH . After grant funding ceased, NOAH became a volunteer-driven project\u2014Kris managed the Spanish content, as well as volunteering to work on the redesign committee so that the new interface was user-friendly to Spanish speakers. For that work, she was one of the awardees when NOAH was given the Thomson Scientific/Frank Bradway Rogers Information Advancement Award in 2006.Journal of the Medical Library Association. In 2021, Kris was selected as a Fellow of MLA.MLA has benefited from Kris's service. She has been a member of the Academy of Health Information Professionals (AHIP) since 1997. She served on the National Program Committee three times and has been elected to the Nominating Committee twice and to the MLA Board. As a member and eventual chair of the Public Health and Health Administration Section (now Caucus), Kris worked with a committee to create a comprehensive list of Medical Subject Headings (MeSH) that would benefit searching for the public health community; many of these terms have been added to the MeSH vocabulary. She also chaired the Research Caucus and served on the editorial board of the I look forward to Kris Alpi's presidential year. Her commitment to professional development and to the growth and evolution of MLA will benefit all members. Please join me in welcoming her to her new position."} +{"text": "The following information is missing from the Funding statement: HK was supported by the Federal Ministry of Education and Research Germany (BMBF) within the project PROVID (FKZ 01KI20160C). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The affiliation for the first author is incorrect. The correct affiliation is not indicated. Wang Jiatong is not affiliated with #1 but with: Postdoctoral College of Teacher Education, Zhejiang Normal University, China."} +{"text": "There are errors in the Funding statement. The correct Funding statement is as follows: This study was supported by the Open Project of Guangdong Provincial Key Laboratory of Tropical Disease Research (G818330002). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The National Institute on Aging (NIA) at the National Institutes of Health, Department of Health and Human Services, is the federally designated lead agency on aging research and supports significant research on aging as a lifelong process. In the last six years, NIA has experienced a tripling of its budget. Although much of this funding is targeted to Alzheimer\u2019s disease (AD) and AD-related dementias research, there has been an increase in funds allocated to non-AD research in keeping with the overall growth of NIH. This symposium will provide a forum for exploration of the implications of the budget increases for the general research community. NIA\u2019s senior staff will discuss research priorities and programs supported by the Institute. A question-and-answer session will follow these remarks on current funding and future priorities and research directions of NIA."} +{"text": "In the original article, we neglected to include the funders, \u201cKey Medical Discipline Construction Project of Tianjin, Science and Technology Project of Tianjin Medical Health Commission (TJWJ2021MS001), Tianjin Key Research and Development Plan, Key Project of Science and Technology Support (20YFZCSY00010).\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "This presentation describes the unique collaboration between The John A. Hartford Foundation, the Administration or Community Living (ACL), and the National Academy for State Health Policy (NASHP) in supporting the RAISE Act Family Caregiver Resource and Dissemination Center, and the goals and activities of the RAISE Act Family Caregiving Advisory Council. Most importantly, she will present the the development of recommendations for a national strategy to support family caregivers involving all levels of government as well as private-sector actors. These recommendations fall into five primary areas, which Fox-Grage will discuss in detail. She will also discuss the Center\u2019s development of family caregiving resources for state and federal policymakers and other stakeholders as well as next steps in turning the Council\u2019s recommendations into concrete action."} +{"text": "The original version of this article unfortunately contained a mistake in Funding section. The correct Funding section is given below.This publication is part of a project that has received funding from the European Research Council (ERC) in the form of an advanced investigator award to SHR under the European Union's Horizon 2020 research and innovation programme (Grant Agreement No. 787270). The publication was also supported in part by a Grant to OMEA from the European Research Council (311723-GENEPAD) and a Grant to SHR from Arthritis Research UK (19799)."} +{"text": "Several factors have been proposed for the geographic variation in the impact of the virus including differences in population age distribution, underlying host genetic and immunological mechanisms . The DELTAS Africa Initiative is an independent funding scheme of the African Academy of Sciences (AAS)'s Alliance for Accelerating Excellence in Science in Africa and supported by the New Partnership for Africa's Development (NEPAD) Planning and Coordinating Agency with funding from the Wellcome Trust (grant No. 107752/Z/15/Z) and the UK government.There are no conflicts of interest."} +{"text": "There is an error in the Funding statement. The correct number for Natural Science Foundation of Guangdong Province is no. 2018A030313610, and the correct number for National Natural Science Foundation of China is no. 81872409.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The authors wish to make the following corrections to this paper :The Funding and Acknowledgments sections need to be corrected. The updated information is provided below:Funding: The team would like to express their thanks to the Deputyship for Research & Innovation, \u201cMinistry of Education\u201d in Saudi Arabia for financial support through the project number IFKSURG-1438-089. Acknowledgments: We thank technical facilitation offered by laboratories of Xianbao Wang and Hongrong Li for support in experimental work. Moreover, the team author extends their appreciation to the National Key R&D Program of China. This change does not affect the scientific results or conclusions in the original published paper.The authors would like to apologize for any inconvenience caused to the readers by these changes."} +{"text": "There is an omission in the Institutional Review Board Statement and Conflict of Interest statements of the paper . The autInstitutional Review Board Statement: Mouse care and experimental procedures were performed under pathogen-free conditions in accordance with approved protocols from the institutional animal care and use committee of the City of Hope National Medical Center (City of Hope IACUC protocol # is 13053 approved 2 November 2014).Conflicts of Interest: D.A.H. and L.E.O. are cofounders of Novonco Therapeutics Inc., Los Angeles, CA, USA. The other authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analysis, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.We apologize for this error and state that the scientific conclusions are unaffected. The original article has been updated."} +{"text": "In the original article, there was an error in the Funding statement. \u201cMT\u201d should be removed from \u201cBTI MT.\u201d The corrected Funding statement should read:meimei@bioasis.us), a grant from the W. Garfield Weston Foundation (RR161038), and donations from the Sullivan Urology Foundation at Vancouver General Hospital. TA was supported by NIH grants 1S10OD010756\u201301A1 and 1S10OD018124\u201301A1. BE was supported by Postdoctoral Fellowships from the Michael Smith Foundation for Health Research, the Centre for Blood Research at the University of British Columbia and the Pacific Alzheimer's Foundation. CS was supported by an Alzheimer's Drug Discovery Foundation Outstanding Young Investigator Scholarship, and by a William and Dorothy Gilbert Graduate Scholarship in Biomedical Sciences at the University of British Columbia and a scholarship from the Centre for Blood Research Graduate Student Award at the University of British Columbia.\u201d\u201cFunding for this work was provided to WJ in the Michael Smith Laboratories, at the University of British Columbia and the Vancouver Prostate Centre, at Vancouver General Hospital, by a grant from the Canadian Institutes for Health Research (MOP-133635), a grant from the Natural Sciences and Engineering Research Council of Canada (CRDPJ 452456\u201313) in collaboration with Bioasis Technologies Inc. (BTI, Additionally, there was a mistake in the COI section, which should now read:\u201cThe authors declare that this study received funding from Bioasis Technologies Inc. The funder did not influence the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication.\u201dThe authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "In the original article, we neglected to include the funder \u201cthe National Natural Science Foundation of China and the National Basic Research Program of China (973 Program), No. 81274000 and No. 2010CB530600 to Fengmei Lian\u201d.Furthermore, there was a mistake in the legend for \u201cThe correct legends appear below.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The lecture will be given by the 2020 recipient, Sara Czaja, PhD, FGSA of Weill Cornell Medicine. The 2020 M. Powell Lawton Award recipient is David Roth, Phd, FGSA, of Johns Hopkins University. The M. Powell Lawton Award is presented annually to an individual who has made outstanding contributions from applied research that has benefited older people and their care. The Lawton Award is generously funded by the Polisher Research Institute of the Madlyn and Leonard Abramson Center for Jewish Life."} +{"text": "People with disabilities face a diverse array of health care and support needs. These needs can vary by disability type, degree, and timing of the advent of functional limitations. These differences have implications for needed health care service use and related expenditures. The symposium will open with a Centers for Disease Control and Prevention-sponsored analysis of adult disability-associated health care expenditures, both nationally and by U.S. state, in total, by per adult, by per adult with disability, and by payer, to illustrate the contribution and variation of these expenditures to individual states and the health care system. We will next present a U.S. Department of Health and Human Services\u2019 Office of the Assistant Secretary for Planning and Evaluation effort to identify onset and patterns of reduced functional ability at end of life for older adults with and without dementia as related to other comorbidities. The last paper will present a Commonwealth Foundation study on older adults with functional disabilities and multiple chronic conditions, comparing those with high health care needs versus the subset of those people who are also high cost. Patterns of utilization differed between these two groups, and by state. These findings have implications for the development of care models that might best meet people\u2019s needs. Our discussant will respond to the studies\u2019 findings and discuss the important role that efforts to understand the nature of disability and functional status and the scale and scope of service use and costs have for people with disabilities."} +{"text": "The authors wish to make the following correction to this paper :The original version of our article did not include the complete funding acknowledgement. The authors wish to change the information in the Funding section from:Funding: This research was funded by the United States Institute of Education Sciences, grant number R305A180290.to:Funding: The research reported here was supported by the Institute of Education Sciences, U.S. Department of Education, through grant R305A180290, to the University of Nebraska-Lincoln. The opinions expressed are those of the authors and do not represent views of the Institute or the U.S. Department of Education.The authors apologize for any inconvenience."} +{"text": "In the original article, we neglected to include the following funding information:\u2018Ministry of Education, Humanities and Social Sciences Research Youth Project, China, 18YJC190023.\u2019The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "There are errors in the Funding statement. The correct Funding statement is as follows: CURACLE Co, Ltd., provided support for the study in the form of salary for HZ, and provided the CU06-1004 sample. The specific roles of this author are articulated in the \u2018author contributions\u2019 section. This research was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government , and the Ministry of Science, ICT & Future Planning . This work was also supported in part by the Brain Korea 21 (BK21) PLUS program. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Bioengineering and Translational Medicine \u2010mimetic hydrogels, which support cell encapsulation to enable regenerative medicine applications.For her scientific achievements, Prof. West has received numerous awards and honors. She is a member of the National Academy of Engineering and the National Academy of Inventors, and a Fellow of the American Institute for Medical and Biological Engineering (AIMBE) and of the Biomedical Engineering Society (BMES). West is a Howard Hughes Medical Institute Professor and in 2015 she received the Clemson Award from the Society for Biomaterials, which is bestowed upon those who have advanced knowledge of material/tissue interactions. In 2017, she was an invited lecturer at the annual meeting of the President's Circle, an honorary association that engages with the Presidents of the National Academies of Science, Engineering, and Medicine to ensure they have the necessary resources to further their work.In addition to her scientific accolades, Prof. West is also known for her outstanding teaching and mentoring. At Duke University, West has been honored with the 2019 Dean's Award for Excellence in Mentoring and with the 2016 Capers and Marion McDonald Award for Excellence in Teaching and Research.These sentiments were echoed by other alumni of the West Lab who were contacted for comment. They described Prof. West as \u201can exemplary leader and mentor\u2026who is always willing to share her experience and knowledge to guide her trainees\u201d and as someone who \u201cnot only promotes the accomplishments of her own trainees but is more broadly dedicated to championing people and raising others up. She is an exemplar of generosity and positivity.\u201d Many noted that Prof. West's guidance extends well beyond the years her students work in the lab, and that she is unwavering in her support. One alumna stated, \u201cIt was her confidence in me that enabled me to develop confidence in myself as a scientist.\u201d On behalf of all of Prof. West's former and current mentees, I thank her for her moral support, guidance, and advocacy, which has positively impacted numerous individuals and contributed greatly to their success.Finally, Prof. West has performed outstanding service to the profession. She has served on the editorial board of numerous scientific journals and participated in several grant review panels for the NIH and NSF. She was Treasurer of BMES from 2011 to 2015 and served on the BMES Diversity Committee from 2014 to 2017. Prof. West was Vice Chair and Chair Elect of the Gordon Research Conference on Biomaterials and Tissue Engineering from 2015 to 2019 (Figure\u00a0Prof. West has had an immense impact on biomaterials and nanomedicine research, and on the field of biomedical engineering more broadly. She is a remarkable leader who has nurtured the careers of many in the field, including both former trainees and those not affiliated with her lab. On behalf of the scientific community, I am honored to thank her for her research contributions, selfless service, and dedication to inspiring and empowering others through meaningful mentorship.Emily Day: Conceptualization; writing\u2010original draft; writing\u2010review & editing.https://publons.com/publon/10.1002/btm2.10225.The peer review history for this article is available at"} +{"text": "In the published article, there was an error in the Funding statement. The correct Funding appears below.\u201cThis work was supported by the National Natural Science Foundation of China (Grant No. 81603204) and National Science and Technology Major Project of China (Grant No.2020ZX09201-009)\u201d.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "This special issue comprises papers based in a meeting of the Nuclear Dynamics Special Interest Group of the Society for Experimental Biology. The session was entitled \u2018Functional organisation of the nuclear periphery\u2019 and was held at the Society\u2019s Annual Main Meeting in the beautiful city of Firenze (Florence) in July 2018. Organized by Katja Graumann and David Evans from Oxford Brookes University, Oxford UK and Roland Foisner, Medical University Vienna, Austria, the session highlighted novel research in plant, animal and fungal nuclear biology since the previous meeting of the group in Brighton, UK in 2016[https://www.cost.eu/actions/CA16212/). A review article in this special issue of Nucleus by members of Workgroup 1 of the COST Action provides insights into microscopy methods for studying 3D nuclear architecture in plants and accompanying challenges being addressed in the consortium[C. elegans[The meeting was supported by attendance of members of the INDEPTH (Impact of Nuclear Domains On Gene Expression and Plant Traits) COST Action CA16212 (onsortium, the cononsortium, and theonsortium with a f. elegans. The fin. elegans providesAs editors, we are grateful to all the contributors, both to the meeting and to this special issue. Their enthusiasm and willingness to share their insights and expertise with others was reflected in their presentations and is evident in the resulting papers. The cross-kingdom format of these meetings provides a fertile environment for generating research ideas and for development of collaborations and this is evidenced in the progress made by the members of the Group."} +{"text": "In the original article, the following was accidently omitted from the acknowledgements section: JB and RLM are members of SPECTRUM a UK Prevention Research Partnership Consortium. UKPRP is an initiative funded by the UK Research and Innovation Councils, the Department of Health and Social Care (England) and the UK devolved administrations, and leading health research charities. This has now been corrected. The title of Emily Thomson was also missing from the author designation box section. Her title of Medical Student has now been added."} +{"text": "Family Physician Dr Cherifa Sururu died of Covid-19 related complications at Mater Dei Hospital, Bulawayo, on Wednesday 27 January 2021.Cherifa was born on 18 March 1970 in Karoi district, Mashonaland West Province, Zimbabwe. His parents originally came from the Yao people of neighbouring Malawi, so he was brought up a Muslim, speaking Yao at home, but later became fluent also in Shona, Ndebele and English. He grew up on a commercial farm, one of 11 children, where his mother was a farm worker and the third wife of his father. His schooling in Karoi and later A-levels in Harare were made possible by a series of scholarships including from the Islamic Development Bank and other well-wishers. He contributed to the family income by joining with seasonal agricultural activities at commercial farms, and supplemented their food supplies with fishing and hunting.Cherifa joined the University of Zimbabwe to study medicine and qualified with MBChB in 1997. He did his house jobs (internship) at Mpilo Central Hospital (1998\u20131999) and then embarked on a long career as a private general practitioner (GP) in Bulawayo. The entrepreneur spirit that saw him selling vegetables, sugar cane and wire toys as a young person, guided him in setting up several successful private surgeries in Bulawayo.Cherifa was a founding member, trustee and second Chairperson of the Islamic Medical Association in Zimbabwe, over a 12-year period. He was active in the College of Primary Care Physicians of Zimbabwe (CPCPZ) where he was the National Treasurer 2006 to 2014 and Honorary National Secretary 2014 to 2017. He was also the CPCPZ representative for the Zimbabwe Medical Association (ZiMA) Matabeleland Branch for 2015 and 2016; Branch Vice President from 2016 to 2017; member of the Social Responsibility Committee from 2014 and organized several medical outreaches to underserved communities in districts. He represented the CPCPZ in the Medical & Dental Practitioners Council of Zimbabwe from July 2020 to his time of death. In the international arena he was the Zimbabwe country representative for Primafamed, a Primary Care and Family Medicine network in Africa, and through CPCPZ, a member of WONCA the World Organization of Family Doctors.Cherifa was one of four private GPs who enrolled with the University of Stellenbosch Family Medicine MMed programme, with the ambition of setting up a similar programme in Zimbabwe. He gained his MMed in March 2017. A register of Specialist Family Practitioners was established at the Medical and Dental Practitioners Council of Zimbabwe in 2015 and the four pioneers of Family Medicine in Zimbabwe were the first to be entered into that register. Family Medicine MMed training programmes were established at the Faculty of Medicine, National University of Science and Technology (NUST), and at the University of Zimbabwe College of Health Sciences, which enrolled their first intakes in 2020 and for whom Cherifa was a committed educator.Cherifa was involved in philanthropic work through Green Crescent Zimbabwe in partnership with the career guidance and counselling department of the Ministry of Primary and Secondary Education, to support a schools\u2019 programme to prevent drugs and substance abuse, teenage pregnancies and suicide. The programme was launched in December 2020. At the time of his death, Cherifa was involved in research on factors influencing substance abuse among school children in Bulawayo and on quality of care in the Zimbabwe district health system.Cherifa felt strongly connected to his parents\u2019 country of origin and managed to visit Malawi in 2017 as part of a Training for Clinical Trainers in Family Medicine run by Stellenbosch University. As a child, he loved listening to the stories about Malawi told by his grandmother by the fireside at night.His death is a sad loss to Family Medicine in Zimbabwe and he will be deeply missed. He is survived by his wife Elizabeth Chipendo and his five children."} +{"text": "Scientific Reports 10.1038/s41598-021-99908-3, published online 14 October 2021Correction to: The original version of this Article contained an error in the Acknowledgments section.\u201cThe authors would like to thank the Henan Adverse Drug Reaction Monitoring Center for providing the data. The data in this study is publicly available and accessible. The information in this paper does not represent the opinion of Henan Adverse Drug Reaction Monitoring Center or national centres. The National Natural Science Foundation of China (No. 72074088) and the Young Scientists Fund (No. 72804052) supported this research. We would like to thank the National Natural Science Foundation of China and the Young Scientists Fund for the funding for this research.\u201dnow reads:\u201cThe authors would like to thank the Henan Adverse Drug Reaction Monitoring Center for providing the data. The data in this study is publicly available and accessible. The information in this paper does not represent the opinion of Henan Adverse Drug Reaction Monitoring Center or national centres. The National Natural Science Foundation of China (No. 72074088) and the Young Scientists Fund (No. 71804052) supported this research. We would like to thank the National Natural Science Foundation of China and the Young Scientists Fund for the funding for this research.\u201dThe original Article has been corrected."} +{"text": "The project title cited in the first sentence of the Acknowledgements is incorrect. The correct title is:\u201cDynamics of Economy and Finance from the Economic Network Point of View\u201d and not \u201cMacro-Economy under COVID-19 influence: Data-intensive analysis and the road to recovery.\u201dwww.editage.com) for English language editing.The correct Acknowledgements should read: This study is conducted as a part of the Project \u201cDynamics of Economy and Finance from the Economic Network Point of View\u201d undertaken at the Research Institute of Economy, Trade and Industry (RIETI). The authors are grateful for helpful comments and suggestions by Discussion Paper seminar participants at RIETI. The authors appreciate the support by INTAGE Inc. to make the data available. The authors would also like to thank participants of the conferences of Computational Social Science Japan, Japan Institute of Marketing Science, and Japan Society for Evolutionary Economics for their helpful discussions and comments. We would like to thank Editage ("} +{"text": "There are errors in the Funding statement. The correct Funding statement is as follows: The authors received no specific funding for this work.The US National Institutes of Health and the Wellcome Trust did not provide any funding for this research and any such claim was made in error.There is an error in the Ethics statement. The correct Ethics statement is as follows: The maintenance of animals and the experimental procedures used in this research program followed the Animal Care and Use Protocol which is approved by the Institutional Animal Care and Use Committee of the Institut Pasteur de Tunis, Tunisia (2018/01/I/ES/IPT/V0). Infected dogs used in this research program were obtained from a previous industry sponsored study that was approved by the Institutional Animal Care and Use Committee of the Institut Pasteur de Tunis, Tunisia (IPT/UESV/27/2012). The Institut Pasteur de Tunis complies with the European Directive for the Protection of Vertebrate Animals used for experimental and other scientific purposes (2010/63/EU)."} +{"text": "In the abstract, in the methodology section, line number 1, the following sentence: \u2018This study was conducted prospectively at the Gezira Hospital for Renal Disease and Surgery and at the National Cancer Institute at the University of Gezira, Sudan, for an 11-year period\u2019 should be replaced with \u2018This study was conducted retrospectively at the Gezira Hospital for Renal Disease and Surgery and at the National Cancer Institute at the University of Gezira, Sudan, for an 11-year period.\u2019"} +{"text": "In the original article, there was errors regarding the biographical data of \u201cHelen S. Mayberg, Zelma Kiss and Andrea K\u00fchn.\u201dNeuropsychiatric Surgery, Helen S. Mayberg, Neurologist, paragraph 1:A correction has been made to the section Helen S. Mayberg is a neurologist born in 1956. She received a Bachelor of Arts degree in psychobiology from the University of California, Los Angeles at the age of 20 and a Medical Doctor degree 5 years later from the University of Southern California. After this, she obtained her certification as a neurologist from Columbia University in New York and a research fellowship in Nuclear Medicine at Johns Hopkins University. Among her multiple honors outstand her election as a member of the National Academy of Medicine of the United States of America, the American Academy of Arts and Sciences, and the National Academy of Inventors of the USA. Currently, she is Director and Professor at The Center of Advanced Circuit Therapeutics at the Mount Sinai Hospital in New York.Neurotechnology, Zelma Kiss, Neurosurgeon, paragraph 1:A correction has been made to the section Zelma Kiss is a neurosurgeon born in 1964. She graduated from the medical faculty of the University of Ottawa, Canada at the age of 24. She completed her neurosurgical training at the University of Toronto, where she also received her Ph.D. After winning the Van Wagenen fellowship, she had the opportunity to continue her postdoctoral education under the supervision of Alim Louis Benabid in Grenoble, France. In 2000, she was appointed at the University of Calgary, where she is currently an associate professor of the Department of Clinical Neurosciences and is the Head of the Neuromodulation program of southern Alberta.Neuroimaging, Andrea K\u00fchn, Neurologist, paragraph 2:A correction has been made to the section In 2010, upon her return to Berlin, she worked as a neurologist at the University Hospital Charit\u00e9 where she became head of the Movement Disorders and Neuromodulation Unit. Her primary research interest became pathological oscillatory activity in patients with movement disorders under deep brain stimulation (Barrow et al., The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "In the original article, there was an error in the Funding statement as published. Instead of \u201cSchool-level special project of Sichuan Normal University,\u201d the correct name for the Funder is \u201cSichuan Normal University's School-level Special Funding Project for \u201cResearch and Interpretation of the Spirit of the Fourth Plenary Session of the 19th Central Committee of the Communist Party of China\u201d \u201d. The funder \u201cNational Social Science Foundation of China (21BGL189)\u201d has also been removed from the statement. The corrected Funding statement is shown below.The work described in this manuscript was supported by the Sichuan Normal University's School-level Special Funding Project for \u201cResearch and Interpretation of the Spirit of the Fourth Plenary Session of the 19th Central Committee of the Communist Party of China\u201d (SNU19J4Z2019-14), the Sichuan Soft Science Research Project (2019JDR0345), the 2020 Sichuan International Science and Technology Innovation Cooperation Project (2020YFH0160), and the Social Science Planning Project of Sichuan Province (SC19TJ026 and SC21B097).The authors apologize for these errors and state that they do not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "In the original article, there was an error in the Funding statement as published. The correct Name for the Funder \u201cHangzhou Normal University (2020QDL006 and 20JYXK035)\u201d is \u201cthe Starting Research Fund from Hangzhou Normal University (No. 2020QDL006), and the cultivation project of the province-leveled preponderant characteristic discipline in the College of Education of Hangzhou Normal University (20JYXK035)\u201d. The corrected Funding statement is shown below.This research was supported by grants to CW from the National Natural Science Foundation of China (32000780), the Natural Science Foundation of Zhejiang Province (Q21C090027), China Postdoctoral Science Foundation (no. 2018M630655), the Starting Research Fund from Hangzhou Normal University (no. 2020QDL006), and the cultivation project of the province-leveled preponderant characteristic discipline in the College of Education of Hangzhou Normal University (20JYXK035), and grants to YY from the Natural Science Foundation of Jiangsu Province (no. BK20190937) and Natural Science Foundation for Universities of Jiangsu Province (no. 19KJB19005).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "In the original article, we neglected to include the funder \u201cThe National Natural Science Foundation of China, No. 81670735 to FC, The National Natural Science Foundation of China, No. 81400802 and Outstanding Youth Training Project from Shanghai Ninth People\u2019s Hospital the National Natural Science Foundation of China, jyyq 08201607 to HL\u201d.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The authors wish to make the following change to the funding section in their paper . The oriThe authors extend their appreciation to the Deputyship for Research & Innovation, Ministry of Education, Saudi Arabia, for funding this research work through the project number IFKSURP-59.We apologize for any inconvenience caused to the readers by this mistake."} +{"text": "Youth Project of Anhui Natural Science Foundation is 2108085QH372. The corrected funding statement appears below.There is an error in the Funding statement. The correct number for This work was financially supported by the National Natural Science Foundation of China (81530096), Shanghai E-Research Institute of Bioactive Constituent in TCM plan, the Opening Project of Shanghai Key Laboratory of Compound Chinese Medicines (17DZ2273300), Youth Project of Anhui Natural Science Foundation (2108085QH372), Key Program for International Cooperation and Exchange of the National Natural Science Foundation of China (81920108033).There is an error in the article title. The word \u201cderivation\u201d in title should be corrected as \u201cdeprivation.\u201dThe authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "NRF-2019R1A2C1084033. The corrected Funding statement is shown below.In the original article, there was an error in the Funding statement. The correct number for the National Research Foundation of Korea (NRF), funded by the Ministry of Science, ICT & Future Planning is NRF-2019R1A2C1084033 to JJ).This work was supported by the National Research Foundation of Korea (NRF), funded by the Ministry of Science, ICT & Future Planning (grant number The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The affiliation of the corresponding author Sharon A. Tooze to the Institute of Cancer Research, Olso, Norway was incorrectly added. The authors would like to apologize for any inconvenience caused."} +{"text": "In the original article, we neglected to include the funder Humanities and Social Science Project of Hefei University, 18RW12ZDA, to Difei Liu. The updated Acknowledgments section is shown below.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The Great Influenza, John M. Barry describes how the distinguished innovators of American medical education and academic public health traveled to Europe at the end of the 19th century to glean all they could for creating a more advanced model for U.S. schools of medicine the interface between public health and health care provision: the Scandinavian welfare state, the German health insurance model, and the British National Health Service. In contrast, the United States has long held the opposite understanding of the role of the state in health: that health is the responsibility of the individual, not of the state \u2014 is moving forward in adopting the social paradigm outlined in the Ottawa Charter. Three recent developments illustrate how Europe is adopting the model.Enabling Good Health for All \u2014 A Reflection Process for a New EU Health Strategy by the former European Commissioner of Health and Consumer Protection . The cons (HiAP) . The cous (HiAP) .Three, an exemplar for a national determinants-based health policy is the Swedish public health policy, Health on Equal Terms, which was adopted by the Swedish Parliament in 2003 . The ove10 Statements on the Future of Public Health in Europe conference held in Montreux, Switzerland, in November 2006 was \"Politics, Policies and/or the Public's Health,\" and the EUPHA has issued n Europe . Perhaps"} +{"text": "Current Oncology that Dr. Richard J. Ablin, from the University of Arizona College of Medicine and the Arizona Cancer Center, has agreed to join Dr. Phil Gold as co-deputy editor. We welcome Richard, who has served on the editorial board of Current Oncology since 1998 and who is currently co-editor with Phil Gold of our section on Updates and Developments in Oncology.We are delighted to inform the readership of In their respective roles, Phil and Richard will work together, with the help of our section editors, to expand the base of submissions from across Canada and the United States. In this regard, we look to you, our readership, for input about the kind of material that you would like to see\u2014and indeed for original articles for publication in your specific areas of interest and expertise, and for reviews of areas that have experienced rapid progress on the clinical side.Current Oncology and working to further enhance the impact value of the journal.Furthermore, Phil and Richard, together with other members of our staff, will be interfacing with industry and other stakeholders in the oncology community throughout North America while promoting Current Oncology to the next level, thereby filling a niche for not only for specialists in clinical oncology, but also for family physicians who have taken on major responsibilities in meeting the needs of cancer patients. In the process, we trust that Current Oncology will become an invaluable resource for the Canadian oncology community.We anticipate that these exciting changes will bring her2 protein overexpression. Overexpression of her2 is predictive of response to particular therapies, including trastuzumab treatment in the meta-static and adjuvant settings, and updated Canadian guidelines for her2 testing are published in this issue of the journal. Two additional clinical practice guidelines are also included: one on the management of solitary brain metastasis, and another on the value of ifosfamide-based combination chemotherapy in advanced soft-tissue sarcoma. Stephen Sagar, in his role of section editor of Integrative Therapies, introduces a manuscript from Edzard Ernst titled \u201cHomeopathy for cancer?\u201d describing the evolution of the art of homeopathy over the last 200 years or so. And the National Cancer Institute of Canada (ncic) Clinical Trials Group\u2019s article \u201cPhase ii testing of sunitinib: the National Cancer Institute of Canada Clinical Trials Group IND Program Trials IND.182\u2013185\u201d leads off a new section of selected Canadian Centre Activities.The gene for human epidermal growth factor 2 is amplified in approximately 20% of breast cancers, and the amplification is the primary mechanism for"} +{"text": "The following treatise includes review chapters from presentations of distinguished scientists at a symposium held August 16\u201317, 2005, in Portland, Oregon, entitled \"Basic and applied biology of the primate reproductive tract: in Honor of the career of Dr Robert M Brenner\". As a prelude to the 2005 annual meeting of the American Society of Primatologists, outstanding scientists from Europe and America assembled to present the current state-of-research in primate reproductive tract biology and to recognize Dr. Brenner's tremendous impact on this research field. In addition, numerous other peers and former students attended and contributed to the scientific exchange and social festivities. Generous support from the National Center for Research Resources (R13 RR018799) and the Oregon National Primate Research Center (ONPRC), Division of Reproductive Sciences, made this event possible.In 2005, Bob Brenner Figure retired So, Bob, we applaud your extraordinary accomplishments during 40 great years at ONPRC! On behalf of your many colleagues and friends at the Center and around the world, we wish you the best and look forward to your continued activities as Scientist Emeritus."} +{"text": "The funding statement for this article should read as follows:We acknowledge financial support through the German Research Foundation DFG . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The National Integrated Project for Prospective Observation of Non-communicable Disease and its Trends in the Aged (NIPPON DATA) is a series of cohort studies on representative Japanese populations that participated in national surveys in Japan. These studies are unique because their participants are general Japanese adult populations from 300 randomly selected areas throughout Japan. NIPPON DATA80 and NIPPON DATA90 are cohort studies of the National Survey of Circulatory Disorders and the National Nutrition Survey in 1980 and 1990. These long-term cohort studies have provided a large amount of important evidence on the risk factors for non-communicable diseases, particularly cardiovascular diseases, and this information has been used in public health policy making, clinical guidelines, and health education in Japan.1 This is a cohort study of participants in the National Health and Nutrition Survey in 2010. NIPPON DATA2010 also includes original examinations and questionnaires added by our research group, as the successor of the National Survey of Circulatory Disorders. Study participants had also participated in the Comprehensive Survey of Living Conditions in 2010; therefore, the data of this national survey were merged with the NIPPON DATA2010 dataset. Thus, the dataset of NIPPON DATA2010 provides a range of information on the socioeconomic status of a representative Japanese population in 2010.A new cohort study, NIPPON DATA2010, was initiated in 2010.Journal of Epidemiology, 11 studies on the relationships among socioeconomic status and obesity, dietary habits, and health-related behavior and knowledge, which are related to cardiovascular risk factors, are published, including the study profile paper of NIPPON DATA2010. Reductions in heath disparities in Japan are one of the main goals of the National Health Promotion Movement in the Twenty-first Century (the second term).2 We hope that the information discussed in this issue will lead to important and effective measures that reduce health disparities and prevent cardiovascular diseases in Japan and worldwide.In this special issue of the Journal of Epidemiology, for his useful instructions and support.We thank Drs. Nobuo Nishi, Nagako Okuda, Takayoshi Ohkubo, and Aya Kadota for their important contribution in editing this issue, and Dr. Keitaro Matsuo, the Editor-in-Chief of the"} +{"text": "Unfortunately, after publication of this article , it was AcknowledgmentsThis study is supported by a grant from the Chinese 973 project (No.2012CB517702) and four grants from the National Natural Science Fund . The authors are very grateful to Prof. Xue-Ying Li for the advice on statistical analysis.They gratefully acknowledge the contribution of the Department of Clinical Laboratory, Peking University First Hospital."} +{"text": "Overall Abstract: Lawrence J. Seidman was born and grew up in New York City. He obtained a PhD from Boston University in Psychology and stayed in Massachusetts to work for many years in the Harvard affiliated Hospitals, such as the VA-Boston Healthcare System, Massachusetts General Hospital and at his untimely passing, he was Professor of Psychology in the Department of Psychiatry at Beth Israel-Deaconess Hospital and the Massachusetts Mental Health Center. He was about to move to a new phase in his career, assuming a position at Children\u2019s Hospital, Boston. He was a pioneer in the fields of the neuropsychology of schizophrenia, ADHD and related disorders, of using the tools of cognitive assessments and brain imaging to understand the genetic predisposition for serious mental illness, and in the last several years\u2014prediction of conversion to psychosis in individuals at high risk. He contributed to many multicenter collaborations and had several collaborators world-wide, playing an important role in their work. This symposium is conducted in his honor with contributions from key collaborators on different aspects of his work. Drs. Tyronne Cannon and Elaine Walker will both represent the North American Prodrome Longitudinal Study (NAPLS) consortium by reviewing its findings in brain imaging and cognition. Dr. David Braff will review the work of the Consortium on the Genetics of Schizophrenia (COGS) multicenter collaboration, in which Dr. Seidman led one of its sites, and Dr. TianHong Zhang from Shanghai will present current data from the Shanghai-Boston SHARP collaboration on early detection of psychosis. The symposium will be concluded by Dr. Keshavan from Beth Israel-Deaconess, who was a close colleague of Dr. Seidman for the last decade. Together they worked side by side exploring many aspects of early detection of schizophrenia and educating trainees and the public on their findings. He will sum up the legacy of Dr. Seidman to the field and how we can continue it into the future."} +{"text": "The Community Preventive Services Task Force recently posted new information on its website: \u201cPhysical Activity: Built Environment Approaches Combining Transportation System Interventions with Land Use and Environmental Design.\u201d This information is available at Established in 1996 by the U.S. Department of Health and Human Services, the task force is an independent, nonfederal, panel of public health and prevention experts whose members are appointed by the director of CDC. The task force provides information for a wide range of persons who make decisions about programs, services, and other interventions to improve population health. Although CDC provides administrative, scientific, and technical support for the task force, the recommendations developed are those of the task force and do not undergo review or approval by CDC."} +{"text": "The Data Availability statement for this paper is incorrect. The correct statement is: This paper uses unit record data from Growing Up in Australia, the Longitudinal Study of Australian Children. The study is conducted in partnership between the Department of Social Services (DSS), the Australian Institute of Family Studies (AIFS) and the Australian Bureau of Statistics (ABS). The findings and views reported in this paper are those of the author and should not be attributed to DSS, AIFS or the ABS.http://www.growingupinaustralia.gov.au/data/dataaccessmenu.htmlData are available from the Longitudinal Study of Australian Children authors. They are contactable via: Users may apply for individual or organisational licenses. There is a fee for the licenses and release of data. The data analysed in this project were accessed via an organisational license. SRZ is the Data Manager for the Telethons Kids Institute and is responsible for maintaining the organisational deed. DC and CLT signed a Deed of Confidentiality to access these data, which SRZ returned for authorisation to the Department of Social Services. No data were analysed which are not available to licensed users of the data.There are errors in the Funding section. The correct funding information is as follows: This work was supported by a grant from the Australian Research Council (CE140100027). The publisher apologizes for the errors."} +{"text": "The affiliation for the eighth author is incorrect. Shane Crotty is not affiliated with #2 but with #1 La Jolla Institute for Allergy and Immunology, La Jolla, California, United States of America."} +{"text": "Dr. Cornelia L. Trimble is not included in the author byline. She should be listed as the twenty-second author and affiliated with Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America. The contributions of this author are as follows: Conceptualization, methodology and formal analysis.The following information is missing from the Funding section: This study was supported by the Dana Foundation and the Maryland Cigarette Restitution Fund to CT and by the NCI grant 1K23CA85437."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This research was suppported through funding from The Funds for Creative Research Groups of Heilongjiang Province of China (JC2016004), Project of outstanding academic leaders in Harbin (2015RQXXJ018), and National Key Research Program of China (2016YFD0100201-21). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The publisher apologizes for the errors."} +{"text": "In the original article, we neglected to include the funder(s) for this article. We gratefully acknowledge financial support as follows. Research support: the Medical Research Council and the National Institute for Health Research Biomedical Research Centre (NIHR BRC) Cambridge . Authors\u2019 support: PH\u2014NIHR Research Professorship, Academy of Medical Sciences/Health Foundation Senior Surgical Scientist Fellowship and NIHR Cambridge BRC. AY is supported by an NIHR Academic Clinical Fellowship. JD is supported by a Woolf Fisher Scholarship. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way.The original article was updated.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "There is, however, approximately 20 to 25 % of Earth\u2019s higher plant species in this area was established by the Chinese Academy of Sciences in 2014. It is an international scientific research and education organisation managed by the Xishuangbanna Tropical Botanical Garden (XTBG). With financial and personnel support from Chinese Academy of Sciences, SEABRI seeks to substantially improve our understanding and conservation of biodiversity in Southeast Asia by cooperation with all CAS institutes, international agencies and government of ASEAN countries.In order to understand and conserve the biodiversity in Southeast Asia, the Southeast Asia Biodiversity Research Institute and a new species of Didymocarpus (Gesneriaceae) from Vietnam, a new species of Aristolochia (Aristolochiaceae), a new species of Dendrobium (Orchidaceae), a new species of Gastrodia (Orchidaceae), a new species of Hedychium (Zingiberaceae) and two new species of Trivalvaria (Annonaceae) from Northern Myanmar, a new species of Primulina (Gesneriaceae) from southwest China and seven species of Begonia (Begoniaceae) from Northern Vietnam and Southern China. The description of the little known species, Begoniakingdon-wardii (Begoniaceae) in Myanmar was also included. Results of molecular phylogenetics of tribe Neottieae (Orchidaceae) are also reported. Most studies are financially supported by the CAS .This special issue of Phytokeys, entitled \u201cPlant diversity in Southeast Asia\u201d represents a new effort by"} +{"text": "In the original research article, Juan Wang and Zengping Gao were not included in the author list. They have now been included along with the affiliation: School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, China.In addition, Zengping Gao has been included as a corresponding author.Finally, the authors unintentionally did not acknowledge funding from the National Natural Science Foundation of China No.31270400.The authors apologize for these oversights. These errors do not change the scientific conclusions of the article in any way.The original article has been updated.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Medicines Travel Award 2018, we received a total of 41 applications from all over the world, of a very high quality. After preselection from the managing editorial staff of Medicines, scientific decision has been performed by four experts from Asia, Australia, and America and by the editor-in-chief of Medicines from Europe. Therefore, the jury included experts from four different continents.For the Medicines, I am pleased to announce the winner of the Medicines Travel Award for 2018. The travel award was granted to Dr. Amie Steel, a postdoctoral research fellow at the Australian Research Center in Complementary and Integrative Medicine at the University of Technology in Sydney, Australia. The award consists of 800 Swiss Francs to attend any academic conference during 2018.As Editor-in-Chief of Dr. Steel is also Associate Director Research at Endeavour College of Natural Health. In the four years since she has received her PhD, she has published over 100 research contributions, including 75 journal articles, two books, and 20 book chapters. She has attracted a large amount of research funding for complementary medicine (CM) research in the last 12 months, the majority of which is directed towards clinical trials involving medicinal cannabis and other natural health products.Her research interests fit within the broad remit of health services research, with a particular focus on understanding clinical practice and supporting clinical research which is embedded in and reflects daily routine care. She is a member of the Steering Committee for three national CM practice-based research networks in Australia, and she has led the formation of the International Research Collaborative for Naturopathic Academic Clinics\u2014a network of research sites in naturopathic educational institutions which crosses four countries. This research infrastructure supports pragmatic clinical research within CM. She is also a peer mentor on the ARCCIM Research Leadership Programs for both CM and Naturopathy, and is a member of the World Naturopathic Federation Research Committee through which she has also contributed to policy development for the World Health Organization.In recent years, she has turned her attention to the evolving field of implementation science. In particular, she is interested in exploring the most effective way to integrate and implement different forms of knowledge. Within CM scholarship, the concept of the \u201cembodied knowledge of the experienced clinician\u201d is defended and criticized in equal measure, and she would like to explore the possibility of drawing upon the strategies being developed within implementation science to address this challenge within her own topic area.Medicines editorial and publishing teams! Congratulations to Dr. Amie Steel on behalf of the international jury and the"} +{"text": "It has been brought to our attention that one funding project of Ministry of Education, Youth and Sports of the Czech Republic LO1417) was missing in the Acknowledgement section of our published paper [417 was mAcknowledgments: The support of the Czech Science Foundation (14-11782S and 16-21053S), Ministry of Education, Youth and Sports of the Czech Republic (LO1305 and LO1417), and Palack\u00fd University Olomouc (IGA_PrF_2017_020) is gratefully acknowledged.This addendum does not cause any changes to the results and conclusions in the original published paper."} +{"text": "The following information is missing from the Funding section: This publication is partly supported by grant #1 U54 CA221205-01 from the National Cancer Institute. The publisher apologizes for the error. The complete, correct funding information is as follows: This work was supported by a training grant #D43TW009575 from the NIH Fogarty International Center and the National Cancer Institutes. This publication is partly supported by grant #1 U54 CA221205-01 from the National Cancer Institute. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of NIH Fogarty International Center and the National Cancer Institute."} +{"text": "In the Funding section, the second grant number from the Ministry of Agriculture of the Czech Republic is missing and should read: This work was supported by the Ministry of Education, Youth and Sports of the Czech Republic , Czech Science Foundation (grant no. P501-12-G090) and supported by Ministry of Agriculture of the Czech Republic, Project No. MZE \u010cR RO0417 and Project No. QK1710302."} +{"text": "Bioengineering & Translational Medicine, I must express my deep gratitude to the members of our Editorial Advisory Board. Their expertise and experience provide a strong foundation supporting the broad range of bioengineering topics that will be covered in the journal. To highlight this strength, we plan to feature one of our distinguished Editorial Advisory Board members in each issue. In this inaugural issue, we are proud to feature Prof Nicholas A Peppas.On the launch of this inaugural issue of AIChE's and SBE's Nicholas Peppas holds a Cockrell Family Regents Chaired Professorship in Engineering, Medicine and Pharmacy at the University of Texas at Austin, with appointments in the departments of Chemical Engineering and Biomedical Engineering, in Surgery in the Dell Medical School, and in the College of Pharmacy. He received his Diploma in Engineering from the National Technical University in Athens and an Sc.D. from the Massachusetts Institute of Technology in Chemical Engineering. He has also received numerous honorary doctorates, including those from the University of Ghent (Belgium), the University of Parma , the University of Ljublijana (Slovenia), the University of Athens (Greece), the University of Patras (Greece), and an honorary professorship from the Sichuan University (China).Professor Peppas' research has made a transformative impact on drug delivery, which has leaped to the forefront of science and technology. In particular, Prof Peppas' research on mathematical models and novel biomaterials has yielded not only new insights into drug release mechanisms and new technologies to control them, but has also provided a new paradigm for research conducted at the interface of medicine and engineering. His early studies on drug diffusion in polymeric systems provided the much needed quantitative tools to describe drug release,Journal of Controlled Release on mathematical models of solute diffusion.Journal of Controlled Release in its 30\u2010plus year history.In 1987, Prof Peppas published two seminal papers in the Responsive hydrogels represent another area where Dr Peppas' work has made a pioneering and transformative impact, especially for applications in diabetes treatment.Encapsulation of glucose oxidase in hydrogel adds another layer of \u201csmartness\u201d to gel\u2010based systems.Professor Peppas has been recognized with numerous awards, including elections to the National Academy of Engineering (NAE), National Academy of Medicine (NAM), National Academy of Inventors (NAI), National Academy of France, Royal Academy of Spain and the Academy of Athens. He is also an elected fellow of the American Institute of Medical and Biological Engineering (AIMBE), Biomedical Engineering Society (BMES), American Association of Pharmaceutical Scientists (AAPS), American Association of Advancement of Science (AAAS), Society for Biomaterials (SFB), among others. Professor Peppas has been recognized by the NAE Founders Award, the Giulio Natta Medal, the Acta Biomaterialia Gold Medal, AIMBE's Pierre Galletti Award, and the Maurice Marie Janot Award in Pharmaceutical Sciences, among others. His teaching accomplishments have been recognized through the Benjamin Garver Lamme Excellence in Engineering Education Award from the American Society for Engineering Education. In 2016, Prof Peppas was included in the 100 Power List by the medicinemaker, comprising the most influential individuals making medicines.AIChE and SBE have been particularly proud to recognize Prof Peppas with several of their distinguished awards, including Materials Engineering and Sciences Award (1984); Food, Pharmaceuticals, and Bioengineering Award (1992); Nanoscale Science and Engineering Award (2014), Founders Award for Outstanding Contributions to Chemical Engineering (2008); William H. Walker Award (2006); Jay Bailey Award (2006); and Institute Lecturer (2007). In 2008, Prof Peppas was listed among the \u201cOne Hundred Chemical Engineers of the Modern Era by AIChE\u201d on the occasion of the Institute's centennial.As towering as these accomplishments and recognitions are, if we ask Prof Peppas, he would say that his proudest professional achievements are his students. To date, he has supervised more than 875 researchers, including 105 PhDs, over half of whom are now professors at various Universities. His students are advancing various frontiers of bioengineering and biotechnology in their own ways in academia as well as industry.AIChE Journal (2009\u20132012). Professor Peppas' pioneering research, along with his active and inspiring leadership in various professional organizations, has advanced the visibility of drug delivery in the scientific landscape, and has inspired young professionals to follow the suit.Over the years, Prof Peppas has played a central role in advancing the cause of bioengineering in various organizations, especially in AIChE. He was director of AIChE (1999\u20132002); chair of the AIChE's Materials Engineering and Sciences Division (1988\u20131990); director of AIChE's Food, Pharmaceutical and Bioengineering Division (1994\u20131997); and founder of AIChE's Bionanotechnology programming area. Even outside AIChE, Prof Peppas has been extremely active in various professional organizations. He was director of the BMES (2008\u20132011), Chair of the College of Fellows of AIMBE (2006\u20132007), President of the SFB (2003\u20132004), president of the Controlled Release Society (1987\u20131988), and chair of the Engineering section of AAAS (2014\u20132015). He is presently the President of the International Union of Societies for Biomaterials Sciences and Engineering (2008\u20132016). Among many of his editorial responsibilities, he was Associate Editor of the Bioengineering & Translational Medicine and look forward to his continued support.On behalf of AIChE and SBE, I thank Prof Peppas for his strong support in the launch of Dept. of Chemical Engineering Center for Bioengineering University of California Santa Barbara, CA 93106 Email:samir@engr.ucsb.eduSamir Mitragotri Editor\u2010in\u2010Chief"} +{"text": "CDC joins the American Public Health Association (APHA) in celebration of National Public Health Week, April 3\u20139, 2017. Since 1995, APHA has led the observance of National Public Health Week during the first full week of April. The week recognizes the impact of public health on the health of the nation. The 2017 observance focuses on making the United States the Healthiest Nation in One Generation by 2030 by spotlighting the importance of prevention, employing successful strategies for collaboration, and promoting the critical role of a strong public health system. http://www.nphw.org/.In conjunction with this year\u2019s observance, CDC is partnering to promote APHA\u2019s National Public Health Week themes, events, tools, and resources. Additional information is available at"} +{"text": "There is information missing from the Funding section. The correct funding information is as follows: This research was supported by JSPS Research Fellowships for Young Scientists (23\u20132815) and special funding for the promotion of internationalization of research activities by the Japanese Group Dynamics Association to RA. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The grant in the Funding Section is incorrect. The correct funding information is as follows: The authors would like to acknowledge the support provided by The College of Engineering Research Center at King Saud University, for providing funds through the research grant, grant no 438/12, for the current research."} +{"text": "There is an error in the Funding section. The correct funding information is as follows: This work was partially supported by JST CREST, Grant Number JPMJCR14D2, Japan and the Core Research for Evolutional Science and Technology and the Center of Innovation Program, the Japan Science and Technology Agency. This work was also partially supported by Japan Science and Technology Agency KAKENHI Grant Number 15H05707. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The publisher apologizes for the error."} +{"text": "The TNM classification for retinoblastoma, which is developed by the American Joint Committee on Cancer (AJCC), has recently been updated from TNM7 to TNM8. The newest dataset for retinoblastoma from the UK's Royal College of Pathologists (RCPath), released in early 2018, reflects the changes in the TNM classification."} +{"text": "It is with great sadness that we share that Jean St. Germain, our colleague, friend, and integral member of the Medical Physics Department at Memorial Sloan Kettering Cancer Center (MSKCC) for over 50\u00a0yr, passed away on December 7, 2017. She died peacefully and was with her brother, Amos, at the time.Jean St. Germain 1945\u20132017Following completion of graduate study at Rutgers University and a fellowship at Brookhaven National Laboratory, Jean was appointed, in November 1967, as a Fellow in the Department of Medical Physics at Memorial Sloan Kettering under John Laughlin and Garrett Holt. At the end of her fellowship, she was appointed to the faculty and rose to the rank of Associate Attending Physicist, and subsequently, to Attending Physicist. She served as the Corporate Radiation Safety Officer, guiding and presiding over the incredible growth of the institution. She served as an interim chair of the Department of Medical Physics from 2007 to 2010 and subsequently as a Vice\u2010Chair for Clinical and Educational Affairs and Clinical Member. Jean was a licensed medical physicist in New York State and was certified in Comprehensive Health Physics in 1974 by the ABHP and in 1991 in Medical Health Physics by the ABMP. Jean was also appointed a Lecturer, Instructor, and ultimately Assistant Professor of Physics in Clinical Radiology, Weill College of Medicine, Cornell University and served as the Radiation Safety Officer at the NY Presbyterian Weill Cornell Medical Center for more than 35\u00a0yr.Jean's contributions to the field of medical physics and health physics were vast and significant. She has served several professional societies in key leadership roles. She served AAPM as National Secretary, Chair of the Rules Committee, Parliamentarian, founding Chair of the Development Committee, Member of the Governing Board of the AIP, Treasurer of the American Academy of Health Physics, Chair of the Examining Panel\u00a0in Medical Health Physics and Vice\u2010Chair of the American Board of Medical Physics. She has served four terms on the AAPM Board of Directors. In the Greater New York area, she served as the President of the Radiological and Medical Physics Society and served three terms as the President of the Greater NY Chapter of the HPS.Jean was a member of the Scientific Committee (SC) for the National Council on Radiation Protection and Measurements (NCRP) that produced NCRP Report No. 105 on radiation protection of medical and allied health personnel. Jean later served as the Chairman of the SC that produced NCRP Report No. 155 on the management of radionuclide therapy patients. In addition, Jean served as a member of several New York State advisory committees on medical and radiological health. She also served as a special examiner for the New York State Civil Service Commission.Jean received many honors and awards during her career. She was a Fellow of the Health Physics Society and of AAPM. She was presented the Failla Award by the Greater NY Chapter HPS and RAMPS. She received the AAPM Distinguished Service Award in 2001 as well as the Varian Award for best professional paper in the Journal of Applied Clinical Medical Physics in 2004. And in 2015, Jean was presented with the Marvin M. D. Williams Professional Achievement Award by the AAPM. The award recognizes AAPM members for an eminent career in medical physics with an emphasis on clinical medical physics.Jean was an excellent lecturer and teacher. She taught Health Physics and Radiation Safety to generations of medical physicists, radiologists, radiation oncologists, nuclear medicine physicians, radiotherapists, radiologic technologists, lab scientists, and others at MSKCC, Weill Cornell Medical Center and throughout the medical physics and radiological community.Beyond physics, Jean's great passion was music. She took vocal lessons at Julliard and was an operatic soprano soloist who gave many recitals and concerts throughout her life. She was also a regular attendee of performances at the Metropolitan Opera. Her commitment to service extended to her church, St. Joseph's in Yorkville, where she was a Trustee.\u00a0She was also an active member of the National Society of Arts and Letters, serving on the Winston Scholarship Committee, and the Shirley Rabb Winston Scholarships in Voice.Jean considered Marie Curie a heroine as she was the first woman to win a Nobel prize and then became the first person to attain a second Nobel prize. Jean had the pleasure of spending time with Marie Curie's daughter, Eve Curie Labouisse, who had written an extensive biography of her mother.She is survived by her brother, Amos St. Germain, his wife, Susan, and their three children. Jean was particularly delighted with her two grand\u2010nieces, Natalie and Maren, and shared photos of them with her colleagues at every opportunity.www.mskcc.org, click on the \u2018Giving\u2019 tab. For additional information, please contact Wei Lui, MSKCC, Department of Medical Physics, 1275 York Ave., New York, NY 10065 (lui1@mskcc.org).In lieu of flowers, donations may be made to the Memorial Sloan Kettering Cancer Center \u2013 John Laughlin Research & Education Fund, in memory of Jean St. Germain. The website is Jean was an accomplished person of wide\u2010ranging interests and service, with a distinct sense of professionalism and honor, who was engaged with the world and loved by friends and family members. She will be greatly missed."} +{"text": "The following information is missing from the Funding section: The Atlanta BEST Program is supported, in part, by the Laney Graduate School at Emory University."} +{"text": "Scientific Reports7: Article number: 41149; 10.1038/srep41149 published online: 01272017; updated: 07172017.The Acknowledgements section in this Article is incomplete.\u201cThis article is submitted as an observational study in the category of original articles. A small sub-set of information from this article has been presented at the annual congress of the British Association of Paediatric Surgeons\u201d.should read:\u201cThis article is submitted as an observational study in the category of original articles. A small sub-set of information from this article has been presented at the annual congress of the British Association of Paediatric Surgeons. Marian Knight is funded by a National Institute for Health Research (NIHR) Professorship. Benjamin Allin is funded by a National Institute for Health Research (NIHR) Doctoral Research Fellowship. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health. The NIHR had no role in design and conduct of the study; collection, management, analysis, and interpretation of the data; and preparation, review, or approval of the manuscript; and decision to submit the manuscript for publication\u201d."} +{"text": "The following information is missing from the Competing Interests section: The tools and methods described in this manuscript are included in a PCT patent application PCT/PT2009000046, which is owned by University of Algarve and the Center of Marine Sciences (CCMAR). Dina C. Simes and Carla S.B. Viegas are cofounders of GenoGla Diagnostics; Cees Vermeer is founder of VitaK. This does not affect our adherence to PLOS's policies on sharing data and materials."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This work was supported by the Ministry of Education, University and Research (FIRB CAROMICS RBAP11B2SX_008) and by the Interuniversity Consortium of Chemistry Research in Metals in Biological Systems (CIRCMSB), Bari, Italy."} +{"text": "The following information is missing from the Funding section: Genotyping of the GS:SFHS samples was carried out by the Genetics Core Laboratory at the Wellcome Trust Clinical Research Facility, Edinburgh, Scotland and was funded by the Medical Research Council UK and the Wellcome Trust Reference 104036/Z/14/Z).These additional funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The following information is missing from the Funding section: This study was supported by the Faculty of Life Sciences, University of Vienna to PM and KS, and the Austrian Science Fund (FWF P29397) to PM and SW.In the Author Contributions section, all four authors should be listed as contributing to the methodology (CM SW KS PM).The publisher apologizes for these errors."} +{"text": "The health, economic and social impacts of disease outbreaks are persistent and manifold. Lessons learnt from the 2013\u20132016 Ebola virus disease outbreak in West Africa demonstrate how far-reaching the cost of pandemics can be. Beyond the devastating death toll of 11\u2009310,Various studies have assessed the systemic failures in the Ebola response, revealing that only about one third of countries in the world can prevent, detect and respond to public health emergencies. These studies have also shown, among other issues: (i)\u00a0inadequate financing for pandemic preparedness; (ii)\u00a0rigid instruments for emergency response; and (iii)\u00a0slow and costly delivery of aid.To support countries to better understand their capacity gaps in pandemic preparedness and response, in 2016 the World Health Organization (WHO) established the joint external evaluation, a voluntary and collaborative process to assess a country\u2019s capacity under the International Health Regulations (2005) to prevent, detect and rapidly respond to public health threats.Even though the World Bank mobilized US$\u00a01.62 billion to support the countries hardest hit by the Ebola outbreak, these efforts were constrained by the lack of flexible instruments for channelling emergency resources in a timely fashion.The World Bank is supporting countries through corporate-level prioritization of pandemic preparedness and response. The International Development Association of the World Bank Group provides loans on soft terms and grants to the 77 poorest countries in the world. The association is replenished every three years.The World Bank is also engaging in strategic partnerships, for instance through its partnership with the governments of Australia and Japan, to support 13 countries across sub-Saharan Africa, Asia and the Caribbean to develop preparedness plans and coordination mechanisms.The World Bank has also set up a Pandemic Emergency Financing Facility. In June 2017, the World Bank launched this facility to offer surge funding in the form of grants and insurance pay-outs, allowing funds to reach countries and international responders more quickly and effectively, and in so doing, prevent outbreaks from becoming pandemics.The World Bank is also using high-level advocacy to strengthen pandemic preparedness and response. Since 2016, the World Bank \u2013 in partnership with the World Economic Forum and the Bill & Melinda Gates Foundation \u2013 has informed decision-makers at the national, regional and global levels about the importance and urgency of investing in pandemic preparedness. Two simulation exercises (October 2016 and January 2017) have informed pandemic preparedness discussions during the Annual Meeting of Ministers of Finance at the World Bank and chief executive officers in the private sector during the World Economic Forum Annual Meeting in Davos. The World Bank is also leveraging its convening role to mobilize resources for regional projects and initiatives, such as the Regional Disease Surveillance and Enhancement Project for West Africa and the World Bank\u2019s collaboration with the Australian government to implement a regional technical assistance programme to ensure sustainability of health security in east Asia. In addition, the World Bank\u2019s International Working Group on Financing Preparedness proposes ways in which national governments and development partners can collaborate to finance investments that will strengthen country preparedness and response capacities for health emergencies.In February 2018, the World Bank in partnership with WHO, the Africa Centres for Disease Control and Prevention and the Bill & Melinda Gates Foundation will host a regional event to define the core elements of a multisectoral pandemic preparedness plan, including planning and coordination mechanisms. In addition, during the Prince Mahidol Award ConferenceTo ensure the success of pandemic financing preparedness, countries should engage in dialogue on the importance of investing in such preparedness. The World Bank\u2019s early successes need to continue through partnerships to renew and sustain national, regional and global commitment and investments for pandemic preparedness and response."} +{"text": "Professor Matthew Tirrell is the founding Pritzker Director of the Institute for Molecular Engineering at the University of Chicago. He received a BS degree in Chemical Engineering from Northwestern University, and he received a PhD in Polymer Science and Engineering from the University of Massachusetts.Prof. Tirrell began his academic career at the University of Minnesota in Chemical Engineering where he earned the Shell Distinguished Chair in Chemical Engineering and established himself as a leader in the study of the polymer interfaces, adhesion, and self\u2010assembly. In the early 1990s, he was named the Head of the Department of Chemical Engineering at Minnesota and later held the Earl E. Bakken Chair of Biomedical Engineering while also serving as the Director of the Biomedical Engineering Institute. In 1998, he moved to the University of California\u2014Santa Barbara to become the Richard A. Auhll Professor and Dean of the College of Engineering. After a few years as the Arnold and Barbara Silverman Chair of the Department of Bioengineering at the University of California\u2014Berkeley, Prof. Tirrell moved to the University of Chicago to build the completely new Institute for Molecular Engineering. The Institute currently comprises sixteen faculty, with diverse research interests focused on innovative technologies in nanoscale manipulation and design at a molecular scale, with potential for societal impact in such areas as energy, health care, and the environment. Among his many talents, he has a knack for defining important scientific problems and assembling powerful, interdisciplinary groups to go about solving them.AIChE, including the Allan P. Colburn Award, the Professional Progress Award, the Charles M.A. Stine Award, an Institute Lecturer Award, and the William H. Walker Award. He has co\u2010authored over 300 papers and one book, and has supervised over 80 PhD students and 40 post\u2010docs.Prof. Tirrell has been elected to the National Academy of Engineering and has been named a Fellow of the American Academy of Arts and Sciences. He is a Fellow of the American Physical Society and received both the John H. Dillon Medal and the Polymer Physics Prize from its Division of Polymer Physics. He has received several awards from AIChE Journal from 1991 to 2000. His ability to work at the interface of the scientific, engineering, and medical communities both as a researcher and as an administrator has resulted in an atypical breadth of publications and awards. He is world\u2010renowned in two distinct areas of research: polymers at interfaces and peptide amphiphile self\u2010assembly. These two areas are tied together by his ability to quantitatively manipulate, measure, and thoughtfully understand the fundamental structural and biological properties of macromolecules.Prof. Tirrell served as the fifth Editor of the Chronologically, his contributions in the area of polymer physics preceded his work with peptide\u2010amphiphiles. As an assistant professor at Minnesota, he began his career working on polymeric surface phenomena, making key contributions in the area of self\u2010healing polymeric interfaces,Prof. Tirrell made a significant departure from his earlier work in polymer physics when he began exploring the area of bioengineering and translational medicine in the 1990s. Together with Prof. Gregg Fields, he worked on the synthesis of a class of self\u2010assembling peptide\u2010based molecule called peptide\u2010amphiphiles.Recent examples include the use of peptide\u2010amphiphile hydrogels for peripheral nerve regeneration, where the Tirrell group synthesized peptide amphiphiles capable of co\u2010assembly with Type I collagen, forming mechanically stable hydrogels and quantifying the enhanced activity of Schwann cells.As students in Prof. Tirrell's lab, we had the unique perspective of learning about the culture of research in an evolving interdisciplinary environment, where Prof. Tirrell encouraged curiosity and collaboration, creating a deep community of engaged scientists and engineers. In the Tirrell research group, Matt was always faithful to his values as an educator, passing on his sense of respect for science and all the people around him. His adventurous nature and willingness to enter new research areas has inspired dozens of his former students to pursue careers in industrial and academic research.Raymond Tu1, James W. Schneider21Dept. of Chemical Engineering, The City College of New York \u2010 CUNY, New York, NY 100312Dept. of Chemical Engineering, Carnegie Mellon University, Pittsburgh, PA 15213Matt pictured with selected PhD students and post\u2010docs from the 1980s. From his 60th birthday celebration at the AIChE Annual Meeting in Minneapolis"} +{"text": "The above mentioned funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.There are errors in the Funding section. The correct funding information is as follows: This research is part of the CGIAR Research Program on Forests, Trees and Agroforestry and the Global Comparative Study on REDD+, with funding support from the USAID (grant number MTO069018,"} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This work was supported by the State of Sao Paulo Research Foudation , by the National Council for Scientific and Technological Development , and by the Rio Preto Research Foundation , Brazil. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The following information is missing from the Funding section: This work was supported by the Grants-in-Aid for Scientific Research from the MEXT (15K21554 to KY), a Wellcome Trust Career Development fellowship and funding under the MRC-CLIMB initiative (to SKS and DF), and a United States National Institutes of Health Research grant R21CA182822 (to IK). DJL is funded by Wellcome Trust and Royal Society grant WT104125AIA. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "This data article provides spatiotemporal patterns of the foundation of Japanese companies in China. The data for companies in the food manufacturing, wholesaling, and service industries were collected from published lists of Chinese companies founded through the investment of Japanese companies. The data are provided in a matrix heatmap format, a two-dimensional visualization of data using color to represent the magnitude of two variables: year of foundation and area in China where the company is located. Specifications TableValue of the data\u2022The data in a heatmap format concomitantly visualize two company variables (year of foundation and area in China where the company is located), allowing researchers to capture an overview of the trends in each industry, and to compare the trends specific to each industry.\u2022The data can be used by researchers to examine the effects of historical events, geographical features, and Chinese policies on industrial exchanges between countries for Japan and other countries.\u2022The data can aid discussion on the business partnership between China and Japan.1To date, there have been four booms in Japanese companies\u2019 investments in China: in 1985\u20131990, 1991\u20131999, and 2000\u20132007, and after 2008 22.1According to a previous report, the food industry was included in all booms of Japanese companies\u2019 investments in China 2.2The number of Chinese companies founded through investment of Japanese companies was collected from references of the 21st Century China Research Institute 2.3The number of such companies was obtained from the list in the source, and filled into a Microsoft Excel for Mac worksheet as shown in"} +{"text": "There is an error in the Funding statement. The correct number for the National Natural Science Foundation of China is 31660282.In the published article, the following affiliation should be added for Xiaoxi Wang: \u201cCollege of Preschool Education and Special Education, Kunming University, Kunming, China.\u201dAlso, the correct affiliation for Yan Liu should be: \u201cFaculty of Foreign Languages and Cultures, Kunming University of Science and Technology, Kunming, China.\u201dThe original article has been updated.The authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The following information is missing from the Funding section: This work has also received funding from the Medical Research Council (MR/M02010X/1) to MJB. The funders had no role in the in the study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There is an error in the Funding section. The correct funding information is as follows: This research was funded by Columbia University through a donation from the Lavine Family. The funder (Lavine Family) had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The publisher apologizes for the error."} +{"text": "In the Funding section, the grant number from the funder National Natural Science Foundation of China, China is listed incorrectly. The correct grant number is: 81200716. The publisher apologizes for the error."} +{"text": "There is information missing from the Funding section. The correct funding information is as follows: This work was funded by Canada Research Chair funds to LS and Leukemia and Lymphoma Society of Canada Grant (#317359) to LS, the Orsino Chair in Leukemia Research, Marvin and Linda Barnett leukemia Research Fund, Brian Steck Leukemia Research Fund to MDM and an Innovation Grant of the Canadian Cancer Society, and Brain Canada with the financial support of Health Canada (# 705166) to JR. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The publisher apologizes for the error."} +{"text": "The funding statement for this article should read as follows:\u201cWork was funded by the German Federal Ministry of Education and Research , the German Research Foundation , the Max Planck Society and the caesar foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "The following information is missing from the Funding section: This research has been supported by the President\u2019s Emergency Plan for AIDS Relief (PEPFAR) through the U.S. Centers for Disease Control and Prevention. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The following information is missing from the Competing Interests section: This research was presented in part at the 20th International AIDS Conference, Melbourne, Australia, 20\u201325 July, 2014. The findings and conclusions in this report are those of the authors and do not necessarily represent the official position of the U.S. Centers for Disease Control and Prevention. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials."} +{"text": "Daniel Shek was born in Hong Kong. He has one elder sister and four younger sisters. His father died when he was in Grade 8. He obtained his bachelor\u2019s degree in Psychology in 1979 at The University of Hong Kong (HKU). He then worked as a full-time demonstrator and pursued his full-time PhD study at HKU for four years.After he obtained his PhD degree, he taught at City Polytechnic of Hong Kong from 1984 to 1987. He then moved to The Chinese University of Hong Kong (CUHK) teaching social work for 22\u00a0years. In response to the severe lack of studies on quality of life of Chinese people, he has gradually developed his research programs on the well-being of Chinese adolescents and families since 1980s.Daniel Shek is currently Associate Vice President (Undergraduate Programme) and Chair Professor of Applied Social Sciences at The Hong Kong Polytechnic University (PolyU). He is also Advisory Professor of East China Normal University, Honorary Professor of Kiang Wu Nursing College of Macau, and Adjunct Professor of University of Kentucky College of Medicine. To promote holistic development of Chinese university students, he has helped to develop the leadership and service learning components of the new 4-year undergraduate curriculum at PolyU.Professor Shek is a psychologist with research interests in positive youth development, family process, scale development, quality of life, program evaluation, addiction, spirituality, leadership and university social responsibility. He has conducted extensive research on the well-being of Chinese adolescents and Chinese families. In the past decade, he has been leading a research project entitled \u201cP.A.T.H.S. to Adulthood: A Jockey Club Youth Enhancement Scheme\u201d (Project P.A.T.H.S.) which is financially supported by The Hong Kong Jockey Club Charities Trust. In this project, he worked together with his colleagues to develop a positive youth development program for junior secondary school students in Hong Kong, trained potential program implementers, provided support in program implementation and conducted comprehensive evaluation of the program. Because of the overwhelming success of the project, the program has been transplanted to mainland China with the financial support of Tin Ka Ping Foundation.Journal of Youth Studies and Applied Research in Quality of Life, Associate Editor of Frontiers in Child Health and Human Development and past Consulting Editor of Journal of Clinical Psychology. He is an Editorial Board member of Encyclopedia of Quality of Life and Well-Being Research published by Springer, Series Editor of The Quality of Life in Asia published by Springer and an Associate Editor of The Encyclopedia of Family Studies published by Wiley-Blackwell. He is an Editorial Advisor of The British Journal of Social Work and Editorial Board member of several international journals, including Social Indicators Research, Journal of Adolescent Health, International Journal of Behavioral Development, and Journal of Child and Family Studies.Daniel Shek is Chief Editor of Professor Shek has to date published more than 600 articles in international refereed journals. His research project on the adjustment of Chinese midlife people has been rated \u201cexcellent\u201d by the Research Grants Council, Hong Kong. He was awarded the CUHK Research Excellence Award 2007 by The Chinese University of Hong Kong. In addition to his research interests, Professor Shek is passionate about teaching. He was awarded two teaching awards during his stay at CUHK. In 2016, he was awarded the Bronze Award in the category of \u201cEthical Leadership\u201d and the Bronze Award in the category of \u201cSocial Enterprise\u201d at the QS Reimagine Education Awards. In 2017, he was further awarded the Silver Award under the category of \u201cEthical Leadership\u201d and the Silver Award under the category of \u201cSustainability\u201d in the QS Reimagine Education Awards.Professor Shek has served on many high-level governmental advisory committees in the Hong Kong Government, including the Action Committee Against Narcotics, Commission on Youth, Women\u2019s Commission, Fight Crime Committee, Beat Drugs Fund Association, Independent Commission Against Corruption, Mental Health Review Tribunal, and Committee on Child Fatality. Professor Shek served as the past Chairman of the Action Committee Against Narcotics (2009\u20132014) and is currently Chairman of the Family Council, Government of the Hong Kong Special Administrative Region. He has also been Chairman of Heep Hong Society and Society of Boys\u2019 Centres. He was awarded the Bronze Bauhinia Star and Silver Bauhinia Star by the Government of the Hong Kong Special Administrative Region in 2000 and 2013, respectively. He is also a Non-official Justice of the Peace in Hong Kong.Daniel Shek lives in Hong Kong with his wife, Jennifer Lee, and their two children, Moses and Esther."} +{"text": "The following information is missing from the Funding section: This article was supported by National Natural Science Foundation of China . The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The Funding statement is incorrect. The publisher apologizes for this error. The correct statement is: This work was supported by National Institute of Allergy and Infectious Diseases . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Phoca vitulina stejnegeri) and phylogenetic relationship between the seals in Japan and other countries. Our results suggested that Japanese harbour seals possibly consisted of more than two lineages and secondary contact of populations after a long isolation. Furthermore, one of the lineage was made only by Japanese harbour seals (Group P1). The proportion of Group P1 was the highest at the South West and gradually decreased towards the North East of Hokkaido, Japan. On the other hand, the haplotypes do not belonged to Group P1 showed close relationship to the seals in the North Pacific. Based on the fossil record of harbour seal in Japan and the range of sea ice during the Last Glacial Maximum (LGM), Group P1 might have entered Japan before the LGM and became isolated due to the geographical boundary, and gradually extended its range from the South West towards the North East of Hokkaido after the disappearance of the sea ice, while the seals which are not in Group P1 immigrated into Japan from the North Pacific.In this study, we used relatively large number of samples (n = 178) and control region of mtDNA (454bp) to clearify the divergence history of Japanese harbour seals ( Phoca vitulina) is an amphibious mammal that distributes across more than 16,000 km of the northern hemisphere (Phoca vitulina richardsi), the southernmost limit in eastern Pacific .,32.31,32These factors suggest the history of Japanese harbour seals: the haplogroup made up only by Japanese harbour seals (Group P1) might have entered Japan before the LGM and became isolated due to the geographical boundary-sea ice, and gradually extended its range from the South West towards the North East of Hokkaido after the disappearance of the sea ice, while the seals which are not in Group P1 immigrated into Japan from the North Pacific, which are the descendent of the seals in refugia in North Pacific.S1 Table(PPTX)Click here for additional data file.S1 FigFinal 369bp of 255 haplotypes were used after alignment. Colouration for the haplotypes of our data and Stanley et al(1996) are same as (PDF)Click here for additional data file.S2 FigFinal sequences of 356bp 381haplotypes were used after alignment ,33,34. C(PDF)Click here for additional data file."} +{"text": "Information was omitted from the Funding statement. The correct version of the Funding statement is available below.\"This work was supported by the Natural Science Foundation of China and the Fundamental Research Funds for the Central Universities (No.2013121026 and No.2011121052) and Xiamen University 985 Project. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "E/V Nautilus, I would like to further thank the Galapagos National Park Directorate for providing joint collection permit PC-45-15 in collaboration with the Charles Darwin Foundation, and thank the government of Ecuador, the Ecuadorian Navy, and its Oceanographic Institute (INOCAR) for their support and permission to operate in their territorial waters. This publication is contribution number 2189 of the Charles Darwin Foundation for the Galapagos Islands.Regarding the specimens collected by the"} +{"text": "The full name of the funder is missing from the Funding section. The correct funding information is as follows: This work was supported by the Engineering and Physical Sciences Research Council [grant number EP/H024891/1]. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Correction to:Cell Discovery (2015) 1, 15040; doi:10.1038/celldisc.2015.40; published online 8 December 2015In the initial published version of this article, there was an inadvertent omission from the Acknowledgements that the contribution of John P Cooke to this work was funded in part by a Core Facility Support Award from the Cancer Prevention and Research Institute of Texas (RP150611). This omission does not affect the description of the results or the conclusions of this paper."} +{"text": "There are errors in the funding section. The correct funding information is as follows: Support was provided by National Research, Development and Innovation Office of Hungary No. K116506 to I.L., the National Research, Development and Innovation Office of Hungary No. K109109 to R.M. and the New National Excellence Program of the Ministry of Human Capacities to \u00c1.L. The publisher apologizes for the errors."} +{"text": "On behalf of everyone at PR&P, I convey the passing of our Editor\u2010in\u2010Chief, Dr Darrell Abernethy. Darrell has been part of PR&P since its inception, initially as Deputy EiC and since 2016 as EiC. He was a stalwart of the journal and assisted in the recruitment of a strong editorial board and the communication with editors of its supporter journals. From a professional perspective, Darrell had a distinguished career as a clinical pharmacologist and geriatrician. Darrell completed MD / PhD degrees at the University of Kansas, followed by training in Internal Medicine at the University of Miami (Florida) and in Clinical Pharmacology at Harvard Medical School / Massachusetts General Hospital. He had a number of academic positions and also directed an NIH program regarding the effects of drugs in the elderly. His last appointment was with the FDA where he investigated ways to predict adverse effects of therapeutic drugs. Darrell's excellence was recognized by his peers with numerous awards, including the Nathaniel T. Kwit Memorial Distinguished Service Award from the American College of Clinical Pharmacology and the William B. Abrams Award in Geriatric Clinical Pharmacology and the Rawls\u2010Palmer Progress in Medicine Award from the American Society for Clinical Pharmacology and Therapeutics.Darrell contributed back to the discipline at a high level. He was EiC of Pharmacological Reviews and Associate Editor of the Journal of Pharmacology & Experimental Therapeutics as well as Clinical Pharmacology and Therapeutics. This experience was brought to bear during his tenure with PR&P. In addition to his editorial responsibilities and leadership, Darrell was also a leader within the broader pharmacology community being a key contributor to the Clinical Division of the International Union of Basic and Clinical Pharmacology (IUPHAR).Darrell's major pastimes included sailing and music. He was an affable person and a great encourager of his juniors. He will be sorely missed.Professor Andrew J Lawrence BSc(Hons), PhD, FBPhSCurrent EiC of PR&P"} +{"text": "In the original article, we neglected to include relevant animal protocols belonging to Hamamatsu University School of Medicine (H28-068 and H29-083). A correction has been made to Materials and Methods, Animals, the second paragraph. All care of experimental animals was in accordance with the institutional guidelines of University of Washington and Hamamatsu University School of Medicine, and experiments were performed as approved by the Institutional Animal Care and Use Committees at the University of Washington (protocol #4359-03) and Hamamatsu University School of Medicine (protocols H28-068 and H29-083). The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way.The original article has been updated.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Acta Horticulturae Sinica. This retraction was approved by the Field Chief Editor and the Specialty Chief Editor of Frontiers in Plant Science. The authors concur with the retraction and sincerely regret any inconvenience this may have caused to the reviewers, editors, and readers of Frontiers in Plant Science.The journal retracts the 23 February 2016 article cited above. Following concerns regarding the originality of the article, Frontiers conducted an investigation. The investigation determined an unacceptably high level of similarity with an article published by Diao et al. in Acta"} +{"text": "The authors of the original article would liThe updated text should read as follows:We are particularly grateful to Prof. Francis for his valuable feedback on our submission for human samples to the Brain Bank for Dementia, UK. We would like to gratefully acknowledge all donors and their families for the tissue provided for this study. Human tissue samples were supplied by the Brains for Dementia Research programme, jointly funded by Alzheimer\u2019s Research UK, the Alzheimer\u2019s Society and the Medical Research Council, and sourced from the Oxford Brain Bank. The Oxford Brain Bank is also supported by the National Institute for Health Research (NIHR) Units.We are sorry for any inconvenience caused."} +{"text": "BioMed Research International has retracted the article titled \u201cAnalysis of the Influence of Complexity and Entropy of Odorant on Fractal Dynamics and Entropy of EEG Signal\u201d due to cThe authors say the research was approved and conducted at Dr. Ahmadian Clinic, 3/22 St., Third Niroo Havaie Ave., Piroozi Ave., Tehran, in September 2015. This was not stated in the article and we cannot find further details of this clinic or Dr. Ahmadian. An ethical approval document for the project \u201cAnalysis of the Influence of Complexity and Entropy of Odorant on Fractal Dynamics and Entropy of EEG Signal\u201d was provided by the first author, dated July 2, 2015, with the approval number D/A/36990 and signed by Dr. Shahaab Ahmadian. The authors also provided blank consent forms in English and Farsi. The institution asked for the article to be retracted and this is supported by the editorial board. The authors do not agree with retraction."} +{"text": "Dear Sir:The article published by Quispe and others2Malaria has devastating effects on the health and way of life of the people all over the world; however, it can be prevented and treated. In America, cases of malaria have been reduced by 58%, and malaria deaths have decreased by 70% since 2000.To analyze the current situation in Peru compared with the rest of South America, we created a chart using World Health Organization data on cases of malaria and the total population, analyzing the incidences by year (Multidisciplinary strategies"} +{"text": "To the Editor: We thank Dr Sasse for his interest in the conclusions of our rapid communication, in which we used the European Centre for Disease Prevention and Control (ECDC) HIV Modelling tool to estimate for the number and proportion of persons living with HIV who remain undiagnosed in the European Union/European Economic Area [mic Area . In respmic Area . We agre"} +{"text": "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The publisher apologizes for the errors.There are errors in the Funding section. The correct funding information is as follows: This study was financially supported by the National Natural Science Foundation of China (grant 41501219;"} +{"text": "Over the last two decades, the concept of self-regulated learning (SRL) has guided scientific approaches to understand successful learning in different fields. Self-regulatory processes were shown to explain achievement differences in students, and SRL may effectively improve achievement in students of different proficiency levels at the University of Music Carl Maria von Weber, Dresden, Germany, and is a full professor paid by the University of Music Carl Maria von Weber. He has received funding from the Dystonia Coalition . He has received speaker honoraria from the Berlin University of the Arts, Germany , from the Hof Symphony Orchestra and the Association of Musicians High Franconia, Germany (2015), from the International Summer Academy L\u00fcneburger Heide, Germany (2015), from the Saxony State Music Council, Germany (2015), from the Klinik Bosse Wittenberg Alexianergemeinschaft GmbH (2015), and from the University of Music and Performing Arts Vienna (2015). The IMM has received free pharmaceutical samples from IPSEN PHARMA GmbH. The IMM has received funding from a private donor without ties to the psychological field.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The following information is missing from the Funding section: This work was supported by the Postgraduate Pro-rectory of the Federal University of Rio Grande do Norte, Postgraduate Program in Nutrition of the Federal University of Rio Grande do Norte (PPGNUT), and Improvement Coordination of Higher Level Personnel . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The publisher apologizes for the error."} +{"text": "The author noted that major part of Acknowledgements section was inadvertently missing in the original article . The new\u201cI am grateful to Morimasa Takayama M.D., the President of Tokyo CCU Network, the Vice-director of Sakakibara Heart Institute, for his expert insight and advice in the ischemia-associated mechanisms of ventricular arrhythmias in acute myocardial infarction, and to Ken Nagao M.D., the Chairman of the scientific committee, Tokyo CCU Network, Division of Cardiology, Nihon University Hospital, for his great suggestions in the scientific interpretation of the large data from Tokyo CCU Network. I also express a special gratitude to all the member hospitals of Tokyo CCU Network for their great contributions to the registry study of acute myocardial infarction in the Tokyo metropolitan area. Finally, I thank Mr. John Martin for his linguistic assistance\u201d."} +{"text": "Reason for erratum:y-axis of Figure Due to a typesetting error, some information went missing on the The publisher apologizes for this error, and the correct version of Figure The original article has been updated."} +{"text": "The authors of this article would like to request the following replacement to the section ACKNOWLEGEMENTS:We are grateful to Mrs. Jin Xu for her assistance in flow cytometry. This study is supported by the National Natural Science Foundation of China (Grant No. 31270032), the National Basic Research Program (973 Program) (No. 2013CB945604), and SJTU Interdisciplinary Research Grant (YG2012ZD05)."} +{"text": "The following information is missing from the funding statement: This work was partially supported by the UIC Research Open Access Article Publishing (ROAAP) Fund. The funder had no role in study design, data collection and analysis, decision to publish, and preparation of the manuscript."} +{"text": "There was an error in the funding statement. The correct funding statement is: This work was supported by National Natural Science Foundation of China and Tianjin applied-basic & cutting-edge research program (09JCZDJC17700). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "An account is provided of a UK national seminar series on Arts, Health and Wellbeing funded by the Economic and Social Research Council during 2012\u201313. Four seminars were organised addressing current issues and challenges facing the field. Details of the programme and its outputs are available online. A central concern of the seminar programme was to provide a foundation for creating a UK national network for researchers in the field to help promote evidence-based policy and practice. With funding from Lankelly Chase Foundation, and the support of the Royal Society for Public Health, a Special interest Group for Arts, Health and Wellbeing was launched in 2015. Arts & Health: An International Journal for Research, Policy and PracticeThe Journal of Applied Arts and Health. A further marker of the maturing of this field is the publication of the Oxford Textbook for Creative Arts, Health and Wellbeing encouraged the development of this field. The Department of Health (DH) broadly supported this work and in 2007 various policy documents were published to support it has played a substantial role in promoting the value of the arts for health, through its journals, organising and supporting conferences and the annual Arts and Health Awards inaugurated in 2008. In advance of the First International Conference for Culture, Health and Wellbeing in Bristol, 2013(1) To review the field of arts, health and wellbeing research and debate aims, methods and achievements.(2) To advance and deepen theoretical, conceptual and methodological developments in researching the role of the arts in health and wellbeing.(3) to develop strategies for advocacy and knowledge exchange with policy-makers, funders, arts agencies and practitioners.(4) to establish the foundations for a national network for arts, health and wellbeing research and evidence-based practice.Research on arts, health and wellbeing is found across many academic disciplines and creative agencies, but remains fragmented, and stands in a developing relationship with the creative arts therapies Hogan, and the The first task of the seminar series was to reflect on the range of academic engagement within the field of arts, health and wellbeing. This was addressed throughout the seminar series, with attention to the range of methodologies applied in research .The final aim, to establish a UK network for arts and health research was fully met as during the programme, funding was acquired from Lankelly Chase Foundation to establish such a network. On an interim basis the steering group for the seminar series acted as a steering group to support the establishment of a new network.Funding from Lankelly Chase Foundation has been of considerable value in helping to maintain the momentum of the ESRC-funded seminar series and build a UK network of researchers working in the field of arts, health and wellbeing.An innovative feature of the work of the RSPH SIG for arts, health and wellbeing is the organisation of webinars on arts and health topics. The first of these in November 2015 focused on two UK arts on prescription initiatives run by Arts and Minds in Cambridge and Pathways2Wellbeing a spin-out initiative from the University of Hertfordshire.In terms of political engagement, the Special Interest Group is acting as the research partner for the UK All Party Parliamentary Group (APPG) inquiry on Arts, Health and Wellbeing which will run over two years 2016\u201317. The secretariat for the APPG is provided by the National Alliance for Arts, Health and Wellbeing, supported by the London Arts and Health Alliance. Management of the APPG secretariat is provided by Alex Coulter, Director of Arts and Health South West. A webpage for the APPG is maintained on the National Alliance website and details of their activities can be found there.The RSPH SIG for Arts, Health and Wellbeing has made an auspicious start and it represents a very welcome initiative in helping to further establish the place of the creative arts in health care and the promotion of health and wellbeing in the wider context of public health. Membership of the SIG is open to anyone with an active interest in arts and health research and the development of evidence-based practice in this field. Further details of membership can be obtained from RSPH (see endnote 8 below).The Seminar Series was funded by the Economic and Social Research Council [grant reference: ES/J022527/1]. Funding from Lankelly Chase Foundation [grant reference: 10290/13564] has supported the development of a UK national network for arts, health and wellbeing."} +{"text": "On Friday 16 June 2017, Dr Andrea Ammon took up office as the third Director of the European Centre for Disease Prevention and Control (ECDC) following her election by the Centre\u2019s management board earlier the same year. The appointment follows a two-year tenure as acting director during which Dr Ammon steered the ECDC steadily and calmly through a challenging period when besides the Centre\u2019s day-to-day work, expertise and resources were requested for the European preparedness and response to global threats such as the Ebola and Zika virus disease outbreaks in Africa and the Americas [Dr Ammon joined the newly established ECDC already in May 2005, as one of its first employees and Head of the Surveillance Unit . While sFrom April 2011 to April 2015, Andrea Ammon was Deputy to the Director and ECDC\u2019s Head of Unit for Resource Management and Coordination. She was a member of the scientific committee for the European Scientific Conference on Applied Infectious Disease Epidemiology (ESCAIDE) and in 2014 and 2015, she headed the committee.A medical doctor by training, Dr Ammon discovered her passion for public health early in her career and she has extensive experience in working in public health authorities at differing levels. Starting at the local and then regional level in the German federal state of Bavaria, she moved to the national public health institute, the Robert Koch Institute (RKI) in 1996, where she was among the first national Field Epidemiology Programme trainees and simultaneously a member of the first cohort of the European Programme for Intervention Epidemiology Training (EPIET). At RKI, she became the Head of Department for Infectious Disease Epidemiology and State Epidemiologist for Germany from late 2002 to 2005. Besides coordinating the national outbreak response team for current and emerging infections, she directed the national field epidemiology training programme and coordinated emergency planning for influenza and epidemiological research programmes in infectious diseases. Furthermore, she provided scientific advice for government ministries, Members of Parliament and the public. In 2003, she coordinated the German response to Europe\u2019s first imported case of severe acute respiratory syndrome (SARS). During her time at RKI, Dr Ammon also became a nationally and internationally respected expert in the field of food- and waterborne diseases. Her PhD was on the synergy between epidemiology and microbiology in the prevention and control of food-borne diseases.Dr Ammon\u2019s professional and leadership skills are complemented by other strong characteristics such as a mind open to suggestions, a capacity for motivating staff and an acute sense of fairness.Eurosurveillance journal and its editors benefited from Dr Ammon\u2019s strategic vision and sense for quality between 2007 and 2015, when she was an associate editor and a strong supporter of the journal. She resigned from this position when taking up her post as ECDC Director to mark the editorial independence of the journal from its publisher and its Director.The"} +{"text": "In the Funding section, one of the grant numbers from the funder National Science Foundation is not listed. The correct funding information is as follows: This research is based upon work supported by National Science Foundation Grants DBI-1046052 and DEB-1554181. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The complete Acknowledgements are as follows: The authors are grateful to the National Statistics Office of Mongolia, the Statistics Department of Ulaanbaatar, and the Health Development Center of the Ministry of Health and Sports which provided access to various demographics and health databases. We are grateful to Social Impact and the Millennium Challenge Corporation for their open access databases of household measurements conducted as part of the impact evaluation of the Energy and Environment Projects. We thank Maria Hernandez, Ajay Pillarisetti, Paul Chung, and Alan Hubbard of the University of California, Berkeley and Nick Lam of the University of Illinois at Urbana-Champaign for their advice and generous assistance during the project. We acknowledge Boldkhuu Nanzad of the Ministry of Energy of Mongolia for advocating for our research, and appreciate Berkeley Air Monitoring Group for facilitating financial arrangements. We also acknowledge that the final analysis benefits from comments made by many participants at a workshop presenting preliminary results conducted as part of the impact evaluation of the Energy and Environment Projects. This material is based upon work supported by the National Science Foundation Integrative Graduate Education and Research Traineeship under Grant No. DGE-1144885. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation. LDH was supported by this NSF traineeship during part of the write up and analysis phases of this project. This NSF traineeship support was generously provided through the Berkeley Center for Green Chemistry. During the peer-review process, a modified version of this paper was included as a chapter in a doctoral dissertation at the University of California, Berkeley. The full dissertation, which includes five total chapters and is titled \u201cA breath of fresher air: improving methods for PMThere is an error in the Methods section in the subsection called Estimating health effects. The correct term for \u201cchronic obstructive pulmonary disorder\u201d is \u201cchronic obstructive pulmonary disease.\u201dThere is an error in the third sentence of the Abstract. The word \u201cassesses\u201d should read \u201cassess.\u201d"} +{"text": "After publication of this work , we noteAcknowledgementsThe authors are very grateful to all members of the 1000 Bull Genomes Consortium for provision of data. This paper is part of the collection \u2018ISAFG2015\u2019 . The publication of the papers in this collection was partly sponsored by OECD Co-operative Research Programme: Biological Resource Management for Sustainable Agricultural Systems (CRP). Ben J Hayes\u2019s participation in ISAFG2015 was financed by the OECD Co-operative Research Programme. The opinions expressed and arguments employed in this paper are the sole responsibility of the author and do not necessarily reflect those of the OECD or of the governments of its member countries."} +{"text": "The following information is missing from the Funding section: This study was supported by a National Science Foundation CAREER award (grant 1453549) to JC. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "A funding organization and grant given to the first author were incorrectly omitted from the Funding Statement. The first sentence of the Funding statement should be: \"Work on this research by CES was supported, in part, by Award Number T32HD007109 from the Eunice Kennedy Shriver National Institute of Child Health & Human Development.\""} +{"text": "The Funding section contains incomplete information. The complete funding information is as follows: This work was supported by National Natural Science Foundation of China , Anhui Provincial Science and Technology Major Project (6030701073 to HZ) and Anhui Provincial Education Department (KJ2015ZD12 to YY). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Anhui Siping Food Development Co. Ltd. provided support in the form of salaries for authors YHL and HPL, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the 'author contributions' section."} +{"text": "There are two errors in this article.The affiliation for the 32nd author is incorrect. Mary A. Marovich, is not affiliated with #13 but with #1 U.S. Military HIV Research Program, Walter Reed Army Institute of Research, Silver Spring, MD, United States of America.The following information is missing from the Disclaimer in the Acknowledgements section: This research was accomplished by Mary Marovich while employed at USMHRP. Dr. Marovich is currently with Division of AIDS, NIAID. The opinions expressed in this article are the author's own and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government."} +{"text": "There is an error in the Funding section. The correct funding information is as follows: This work was supported by the Guangxi Key Technologies Research and Development Program (0992033\u20135), Guangxi Government Senior Scientist Foundation (2011B020), and Guangxi Natural Science Foundation (2016GXNSFCA380003). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Missing FundingIn the original article, we neglected to include the funder Medical Research Council, Grant No. G1002277, ID98489 for this study. We neglected to include the Medical Research Council in our list of funders in the Acknowledgements section. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way.Text CorrectionIn the original article, there was an error. We neglected to include the Medical Research Council in our list of funders in the Acknowledgements section.A correction has been made to the Acknowledgements, page 17:via an award to the Cambridge NIHR/Wellcome Trust Clinical Research Facility. MM is funded by the Medical Research Council. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The authors thank T. V. Veenith, P. J. Coles, and D. K. Menon for providing images for Figure 2. The authors also thank D. K. Menon for helpful comments. Our cerebral metabolism studies were funded by the Medical Research Council . The following funding sources should also be acknowledged: PH is supported by a National Institute for Health Research (NIHR) Research Professorship, Academy of Medical Sciences/Health Foundation Senior Surgical Scientist Fellowship and the National Institute for Health Research Biomedical Research Centre, Cambridge. PH and KC are supported by the NIHR Biomedical Research Centre, Cambridge. MS is supported by PH\u2019s NIHR Research Professorship. AS is funded by the NIHR The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The Community Preventive Services Task Force recently posted new information on its website: \u201cDiabetes Management: Team-Based Care for Patients with Type 2 Diabetes.\u201d The information is available at Established in 1996 by the U.S. Department of Health and Human Services, the task force is an independent, nonfederal panel of public health and prevention experts who are appointed by the director of CDC. The task force provides information for a wide range of persons who make decisions about programs, services, and other interventions to improve population health. Although CDC provides administrative, scientific, and technical support for the task force, the recommendations developed are those of the task force and do not undergo review or approval by CDC."} +{"text": "Due to a typesetting error, a funding statement which does not apply to the article was added. The following text has, therefore, been removed from the article:This work was supported by Natural science outstanding youth fund of Shandong Province (JQ201607), Taishan Young Scholar Program of Shandong Province, AoShan Talents Program supported by Qingdao National Laboratory for Marine Science and Technology (No.2015ASTP), \u201c100-Talent Project\u201d of Chinese Academy of Sciences for CS.This error does not change the scientific conclusions of the article in any way. The publisher apologizes for the mistake.The original article has been updated."} +{"text": "Information was missing from the Funding section. The correct funding information is as follows: This project was supported by grants from the National Institute of Allergy and Infectious Diseases (RO1A1073745 and RO1AI080799). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Over the past two decades, genomics has evolved as a scientific research discipline. Genomics research was fueled initially by government and nonprofit funding sources, later augmented by private research and development (R&D) funding. Citizens and taxpayers of many countries have funded much of the research, and have expectations about access to the resulting information and knowledge. While access to knowledge gained from all publicly funded research is desired, access is especially important for fields that have broad social impact and stimulate public dialogue. Genomics is one such field, where public concerns are raised for reasons such as health care and insurance implications, as well as personal and ancestral identification. Thus, genomics has grown rapidly as a field, and attracts considerable interest.One way to study the growth of a field of research is to examine its funding. This study focuses on public funding of genomics research, identifying and collecting data from major government and nonprofit organizations around the world, and updating previous estimates of world genomics research funding, including information about geographical origins. We initially identified 89 publicly funded organizations; we requested information about each organization's funding of genomics research. Of these organizations, 48 responded and 34 reported genomics research expenditures . The figures reported here include all the largest funders and we estimate that we have accounted for most of the genomics research funding from government and nonprofit sources.Aggregate spending on genomics research from 34 funding sources averaged around $2.9 billion in 2003 \u2013 2006. The United States spent more than any other country on genomics research, corresponding to 35% of the overall worldwide public funding . When adjusted to genomics funding intensity, however, the United States dropped below Ireland, the United Kingdom, and Canada, as measured both by genomics research expenditure per capita and per Gross Domestic Product. Genomics research, as a field of study, is largely a creature of the past two decades and is generally defined as the study of whole genomes. The term genomics came into common use in 1987 to distinguish \"high throughput\" data- and technology-intensive approaches to studying DNA structure and function from the more established approach of studying DNA structure and function of individual genes. The history of genomics research is embedded in the Human Genome Project and its parallel private sector components. In 2003, with the completion of a high-quality sequence of the human genome, the National Human Genome Research Institute (NHGRI) of the National Institutes of Health (NIH) announced that \"the genomic era is now a reality\" , Human Genome, and Functional Genomics/Health Effects -36. The Genomics funding from the Centers for Disease Control and Prevention (CDC) increased from US$3.85 million (2003) to US$6.95 million (2006) ,38; partThe US Department of Defense supports research and development through the Defense Advanced Research Projects Agency (DARPA), which pursues research and technology where risk and payoff are both very high , and thrTo estimate the completeness of the United States spending on genomics research reported here, we determined the percentage of federal and nonprofit funding on R&D that is covered by the entities in this survey. Of the US$1,023 \u2013 1,064 million per year in federally funded research and development in all disciplines (2003 \u2013 2006), six of the entities included in this survey accounted for 84 \u2013 86% . AlthougThe European Commission funds genomics research in the European Union, plus Iceland, Norway, Liechtenstein, Switzerland, Israel, and Turkey. In the Sixth Framework Programme of the European Commission, covering 2002 \u2013 2006, the major funding areas that included genomics research were Advanced Genomics and its Applications for Health, and Combating Major Diseases . EuropeIn an effort to include the European countries funded through the European Commission but not otherwise covered in this world survey, we contacted organizations from Austria (Fonds zur Forderung der Wisenschaftlichen Forschung), Denmark , Estonia (Estonian Genome Foundation), Finland (Academy of Finland), France , Hungary , Iceland (Rannis), Italy , Poland (Ministry of Scientific Research and Information Technology), and Sweden , but we received no response from any of these except for the Academy of Finland and Centre National de la Recherche Scientique of Canada, the National Research Council (NRC) and the Canadian Biotechnology Strategy (CBS). The CBS includes six departments and agencies that are building capacity for new research ), althouJapan's spending on genomics research steadily increased between 2003 and 2006, when it reached 5% of worldwide public funding Table . AlthougThe values reported in this world survey for Japan are likely a near-complete representation of the government's investment in genomics, since the four Japanese ministries reported in this survey are the main government funders of life science and genomics research -60. One Our source for China indicated that the central government funded about US$40 million per year of genomics research through the National Natural Science Foundation of China (NSFC), the Chinese Academy of Sciences (CAS), and two programs at the Ministry of Science and Technology (MOST): the National High-tech Development R&D Program (863 Program) and the National Key Basic Research Program (973 Program). The source also indicated that an equal amount of funding was provided by local governments , yielding a total of US$80 million per year \u2013 or about 3% of worldwide public genomics research funding Figure . Since oIn Germany, the Federal Ministry for Education and Research coordinates the R&D initiative for the federal government . AnalysiThe primary funder of genomics research in the Netherlands is the Netherlands Genomics Initiative (NGI), which was established by the Netherlands Organization for Scientific Research and five government ministries in 2001. The NGI was created to combine existing strengths into one initiative composed of core activities and research programs . NetherlIn South Korea, which accounted for 2% of total genomics research funding in 2006 Figure , the MinAccording to the Office of Science and Technology of the Ireland Department of Enterprise, Trade, and Employment, three government agencies are responsible for most of the genomics funding in Ireland . These three agencies \u2013 the Higher Education Authority, the Science Foundation, and the Health Research Board \u2013 are included in this survey. The Irish government has recently increased its investment in R&D with the goal of meeting or exceeding the average R&D investment of European Union countries . Indeed,Genoma Espa\u00f1a is the Foundation for the Development of Genomic and Proteomic Research, and is backed by the Spanish State through the Ministry of Health and Consumer Affairs (Ministerio de Sanidad y Consumo), and the Ministry of Education and Science (Ministerio de Educacion y Ciencia) . Genoma In Australia, the National Health and Medical Research Council (NHMRC) is the \"peak body\" for supporting health and medical research and provThe government agency responsible for supporting basic and applied research in South Africa is the National Research Foundation , while mThe three main funders of genomics research in New Zealand are the Foundation for Research in Science and Technology (FRST), the Royal Society, and the Health Research Council . Only FRST, which represents half of the New Zealand government-funded R&D , respondAs described by the World Health Organization, genomics research is beginning to occur in developing countries and regions such as Brazil, China, India, and the Asia-Pacific Region . We contThis study represents only government and non-profit \"public\" funders of genomics research. The previous survey in 2000 included private companies, and showed a roughly 2:1 ratio of private genomics R&D to public funding . In 20There are two major caveats to consider when interpreting the figures reported in this world survey. First, many organizations and countries are not included in this study, either because we did not know to contact them or because they did not respond. Thus, the full amount of genomics research funded by public sources is certainly higher than the estimates reported in this study.genomics, \"the study of genes and their function\" \" including \"interactions of those genes with each other and the person's environment\" ,79). Intunction\" , that ishe NHGRI ,27 and rAlthough this survey is an estimate of funding for genomics research, and necessarily fuzzy and approximate, it represents the only attempt to perform such a survey to our knowledge, and therefore provides patterns and trends as a rough indicator for planning among researchers, science administrators, policymakers, and the general public.This world survey would not have been possible had we tried to gather funding information for genomics research according to a strict and consistent definition. If governments and private funders believe that genomics as a funding category is permanent and worth retaining as a separate category for analysis, it will need a more uniform definition. This is particularly relevant for the NIH, the world's largest biomedical research funder, which, except for two constituent institutes, (the NHGRI and the NCI) does not track funding of genomics research. And these two Institutes report widely different figures for genomics as a fraction of genetics research, reflecting some real differences but probably also reflecting different definitions of genomics research. Furthermore, the closest research category that the NIH does track, genetics, is not defined centrally.The NIH is in the process of developing a \"portfolio analysis\" web-accessible tool that will allow the public to access information about NIH projects by research area, the definitions of which will be \"laboriously crafted with input from hundreds of scientists\" . The effThe NCI, a component of the NIH, provided a definition of genomics: \"the identification, characterization and quantification of all genes involved in a particular pathway, organelle, cell, tissue, organ or organism that can be studied in concert to provide accurate and comprehensive data about that system\" . This is a detailed definition that encompasses and expands upon the definition of genomics already in use by the NHGRI and the CDC (see above). If the NIH includes genomics as a research area in its new portfolio analysis tool, the NCI definition could serve as a starting point for the development of a trans-NIH definition of genomics research.Genomics research has become incorporated into scientific and medical research, and is beginning to be applied in medicine and commerce. Genomics captures the attention of the general public because of its technological power to study the structure and function of DNA, with the consequent potential to reveal intimate details about individuals, populations, and associations between genotype and phenotype.The amount of funding provided for genomics research is of interest to both scientific and lay communities worldwide. This world survey indicates that overall public funding for genomics research, as a minimum estimate, averaged around US$2.9 billion annually (2003 \u2013 2006). Government and nonprofit funding of genomics research is likely to comprise between one-third and one-half of the total funding (where the remainder is for-profit private funding), based on the worldwide distribution of funding for health research , with the understanding that estimates were acceptable. In some cases, initial contact was by phone. Follow-up email correspondence often occurred, with the most effort being expended on procuring figures from the largest public funders. In a few cases, such as China, we were able to obtain funding information only from intermediaries familiar with science budgets because of their role in national planning and as performers of genomics research.Data were requested by fiscal year and are reported in the tables for the calendar year that encompassed most of that fiscal year. Specifically, the calendar year at the beginning of the fiscal year was used when the fiscal year began on April 1, while the calendar year at the end of the fiscal year was used when the fiscal year began on October 1. When funding amounts were supplied in a currency other than US$, they were converted to US$, using the purchasing power parity (PPP) indices provided by the OECD , except To determine per capita genomics funding, the amount of funding per country (in US$) was divided by the estimated population in the middle of the year, as provided by OECD for 2003\u20132005 . The popRankings listed in the tables were determined by ordering 2006 values, except in the two instances where the 2006 data were not reported. In those cases, an average of the three previous years determined the ranking order for 2006.AAAS: American Association for the Advancement of Science; ATIP: Asian Technology Information Program; BBSRC: Biotechnology and Biological Sciences Research Council (United Kingdom); BMBF: Budesministerium fuer Bildung und Forschung (Germany); CAS: Chinese Academy of Sciences; CBS: Canadian Biotechnology Strategy; CDC: Centers for Disease Control and Prevention (United States); CDMRP: Congressionally Directed Medical Research Program (United States); DARPA: Defense Advanced Research Projects Agency (United States); DFG: Deutsche Forschungsgemeinschaft (Germany); DHHS: Department of Health & Human Services (United States); DHS: Department of Homeland Security (United States); DNA: Deoxyribonucleic Acid; DOD: Department of Defense (United States); DOE: Department of Energy (United States); FAPESP: Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo (Brazil); FRST: Foundation for Research in Science and Technology ; FY: Fiscal Year; GDP: Gross Domestic Product; HHMI: Howard Hughes Medical Institute (United States); IMF: International Monetary Fund; MAFF: Ministry of Agriculture: Forestry and Fisheries (Japan); METI: Ministry of Economy, Trade and Industry (Japan); MEXT: Ministry of Education, Culture, Sports, Science and Technology (Japan); MHLW: Ministry of Health, Labour and Welfare (Japan); MOST: Ministry of Science and Technology (Japan); NASA: National Aeronautics and Space Administration (United States); NCI: National Cancer Institute (United States); NGFN: National Genome Research Network (Germany); NGI: Netherlands Genomics Initiative; NHGRI: National Human Genome Research Institute (United States); NHMRC: National Health and Medical Research Council ; NIH: National Institutes of Health (United States); NIST: National Institute of Standards and Technology (United States); NOAA: National Oceanic and Atmospheric Administration (United States); NRC: National Research Council (Canada); NSERC: Natural Sciences and Engineering Research Council (Canada); NSF: National Science Foundation (United States); NSFC: National Natural Science Foundation of China; NOW: Nederlandse Organisatie voor Wetenschappelijk Onderzoek (Netherlands); OECD: Organization for Economic Cooperation and Development; PPP: Purchasing Power Parities; R&D: Research & Development; TIGR: The Institute for Genome Research (United States); UK: United Kingdom; US: United States; USDA: United States Department of Agriculture.JRP generated the research question, contacted the organizations listed in this survey, generated and analyzed the data, and drafted the manuscript. RMCD participated in study design and coordination, contributed the initial contact list from the 2000 World Survey, and helped to draft the manuscript. Both authors read and approved the final manuscript.Identified organisationsClick here for file"} +{"text": "Public Health in Rajasthan, published in 2007 has 227 pages, divided into 25 chapters and 5 Appendices. Book has attractive title page and is printed on good quality paper with a comfortable font size. Chapters cover wide range of subjects related to National Rural Health Mission, Reproductive and Child Health, Integrated Child Development Services, National programs on Tuberculosis, Leprosy, HIV/AIDS, Blindness, National Vector Borne Disease Control Program, Integrated Disease Surveillance Project, Computerization and Management Information System, Medicolegal aspects, Consumer Protection Act, and Behavior Change Communication for Health, etc. There are two chapters devoted to role of The United Nations Children's Fund (UNICEF) and The United Nations Fund for Population Activities (UNFPA) in Rajasthan. Brief description of the projects undertaken through these agencies is useful. Galaxy of 21 prominent experts have contributed in this book. Each chapter provides a brief description of the state-specific program and has been written to act like an operational guide for the implementation of the program. Authors have nicely and succinctly compiled the information available at various places, and have also given resource references for further reading at the end of each chapter. Editor has very rightly claimed that the book can be labeled as a rapid reader for the doctors working at peripheral facility. This can also be good resource for public health students undertaking Masters of public health (MPH) or Doctorate in Medicine (MD) in the specialty. This would also be a good resource for all States to enable them to draft their own State-specific resource books and to have a ready reference for the programs that are operational in Rajasthan.The book shiv_mathur@hotmail.com.Prof. Shiv Chander Mathur, The Editor of the book is a prominent public health expert with national and international teaching training and research experience. He has published more than 50 papers and two monographs on public health. He has also received Government of India merit award for his writings on primary health care. For any further queries readers may contact the editor. E-mail:"} +{"text": "The funding information for this paper was omitted. The complete funding information should say: This work was supported by the Deutsche Forschungsgemeinschaft (DFG) and the Tinnitus Research Initiative (TRI). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Journal of Experimental and Clinical Cancer Research, the corresponding author has informed the journal that this article had been accepted and previously published by Acupuncture and Electro-therapies Research [Journal of Experimental and Clinical Cancer Research. The authors apologise for any inconvenience this may have caused to the editorial staff and readers.Following the publication of this article in JournResearch . Since i"} +{"text": "There was an error in the fifth author's name and affiliation. The fifth author's name should be Ruben C. Gur and he is additionally affiliated with Philadelphia Veterans Administration Medical Center, Philadelphia, Pennsylvania, United States of America."} +{"text": "County Health Rankings and deputy director of its parent project, Mobilizing Action Toward Community Health (MATCH), funded by the Robert Wood Johnson Foundation. She also directs the Making Wisconsin the Healthiest State project, a research and translation effort funded by the Wisconsin Partnership Program, University of Wisconsin School of Medicine and Public Health. In this interview, Dr. Booske gives an overview of the MATCH articles in the September 2010 issue of Preventing Chronic Disease and talks briefly about the role of incentives in public health.Bridget Booske is the project director of podcast - \"Can Incentives Improve Population Health?\""} +{"text": "To the Editor: Chikungunya is a disease caused by an arboviral alphavirus transmitted to humans by Aedes mosquitoes . Symptoms include fever, myalgia, rash, and joint pain , Europe, and the Middle East reported around the world. A literature review was undertaken on Medline by Pubmed and websites provided by the World Health Organization, Eurosurveillance, European Center for Disease Prevention and Control, Health Protection Agency (United Kingdom), Institut de Veille Sanitaire (France), and the Centers for Disease Control and Prevention (United States) were searched for information on imported chikungunya cases. Data were then mapped and compared with the known and theoretical geographic distributions of he world (4\u20138). TThese facts underscore the need for clinicians to consider the possibility of chikungunya disease in patients who experience acute unexplained fever with joint pain and live in regions where mosquito vectors are established. The presence of imported cases and well-established vectors also confirms the need for an active surveillance system; early detection of unexpected new diseases by physicians will enable the timely implementation of suitable control measures that can interrupt the transmission chain.Aedes albopictus and Ae. aegypti mosquitoes. World repartition of Ae. albopictus mosquitoes (tan areas) and theoretical dispersion of Ae. aegypti in 2008 according to the World Health Organization. Areas where imported cases of chikungunya have been reported during 2005\u20132008 are marked with a purple circle or a purple triangle when the number of imported cases was unknown. Data sources: US Centers for Disease Control and Prevention, World Health Organization, and literature review on Medline by Pubmed (www.esri.com/software/arcgis).Imported cases of chikungunya virus infection and known and theoretical geographic distributions of"} +{"text": "Preventing Chronic Disease, CDC's online journal on public health policy, practice, and research (http://www.cdc.gov/pcd/). The Matthews discussion focuses on the importance and challenges of connecting with private-sector stakeholders and the strategies necessary to develop private networks to assist public health in the 21st century.Gene Matthews, former chief counsel at the Centers for Disease Control and Prevention (CDC) and a senior fellow with the North Carolina Institute for Public Health, was interviewed by Elizabeth Majestic with CDC's National Center for Chronic Disease Prevention and Health Promotion for The interview summarizes the successes of early private-sector partnerships with the public health community. During the past 50 years, however, public health has become more internally focused and less engaged with both the business community and the political process. Matthews suggests that in an atmosphere of doing more with less, public health has to face the rigors of accountability and to think in different ways about strategic partners and how to match purposes ethically.The interview was filmed in November 2008.Public health has a long history of working successfully with the business sector. Initial efforts focused on providing basic health services through a partnership with the Rotary Club and expanded to fighting infectious diseases with a variety of private-sector partners. In the last 50 years, however, as public health has become increasingly isolated from the private sector, achieving public health objectives has been more difficult. Two examples of a new type of business relationship are highlighted from the National Institute of Occupational Safety and Health: 1) the tripartite agreements that called for the unions, company leaders, and CDC staff to work together to improve worker safety and 2) the National Occupational Research Agenda.Urgent threats such as those caused by terrorists tend to unite the public health community with all sectors, including the business sector. When these threats arise, the public and private sectors provide an initial influx of resources. These resources tend to be cyclical, but they can be sustained by forming relationships that address not only the urgent health threats such as bioterrorism, but also chronic diseases.Public health needs to develop new partnership models that take into account modern health problems. By building credibility with business and community leaders on issues such as preparedness, public health can gain support for its chronic disease efforts. Initiatives that are considered desirable by communities, such as those to build recreational facilities, can help limit obesity. Public health leaders need to find this common ground.Underlying the discussion of partnerships and the debates over definitions, motives, and processes are basic questions of ethics. Which partnerships should be considered? How can their accountability be assured? To help public health professionals develop productive and ethical public-private partnerships, Matthews highlights 3 strategies that should serve as the cornerstone for partnership development.The economic downturn will increase the demands on public health. The public health community needs to prepare for the likely budget cuts, report the impact of these cuts on the public's health, and expand the range of stakeholders that influence the body politic. The Department of Agriculture is highlighted as an agency that has met these challenges by creating relationships with the private sector.Gene W. Matthews is a senior fellow at the North Carolina Institute for Public Health, the outreach and service unit of the University of North Carolina School of Public Health. He also holds faculty appointments at the University of North Carolina School of Public Health and the Georgia State University College of Law. Since 1999, Mr Matthews has provided leadership in the development of CDC\u2019s internal Public Health Law Program, an effort to reach out to the legal community and to public health practitioners. In June 2004, Mr Matthews received the Distinguished Career Award of the Public Health Law Association \u201c\u2026 in recognition of a career devoted to using law to improve the public\u2019s health.\u201d Mr Matthews is a graduate of the University of North Carolina School of Law and is a member of the North Carolina Bar."} +{"text": "To the Editor: In a recent letter and ELISA for immunoglobulin G, as were the current Brazilian HPS cases (Rattus rattus and R. norvegicus ) are the only Old World rodents ubiquitous in the New World and thus a potential source of SEOV infection in the Americas (Wild rats (The clinical syndromes of HFRS and HPS can appear identical, with pulmonary edema, shock, and renal insufficiency with marked proteinuria and thrombocytopenia ("} +{"text": "Dr. Pritam Singhth October 2009. Apart from being the President of the ISA in 1970 and the Editor of IJA from 1964 to 1970, he held various prestigious posts like visiting Professor of the University of Cincinnati College of Medicine, Ohio, USA; Professor and HOD of Anaesthesiology at CMC, Ludhiana; Principal of Govt. Medical College, Amritsar and Director of Research and Medical Education of Punjab. He had served the Indian Army Medical Corps from 1942 - 1946 and rose to the rank of CaptainDr. Pritam Singh, ex-president and editor ISA breathed his last at Cincinnati USA on 16Dr. G Subramanianth November 2009 at the age of 70 years. An ISA life member, Dr G. Subramanian was the founder President of Dindigul ISA, TN state. He is survived by his wife and a son.With profound grief, Dindigul City Branch of the TN Chapter ISA reports the sad demise of Dr. G Subramanian on 29"} +{"text": "The World Health Organization (WHO), the US Centers for Disease Control and Prevention, and the Canadian Public Health Association developed the Global Tobacco Surveillance System (GTSS) to assist WHO member states in establishing continuous tobacco control surveillance and monitoring. The GTSS provides a flexible system that includes common data items but allows countries to include important unique information at their discretion. It also uses a common survey methodology, similar field procedures for data collection, and similar data management and processing techniques. The GTSS includes collection of data through three surveys: the Global Youth Tobacco Survey (GYTS) for youth, and the Global School Personnel Survey and the Global Health Professional Student Survey for adults.Countries can use data from the GYTS to monitor and evaluate National Tobacco Control Action Plans and articles from the WHO Framework Convention on Tobacco Control (WHO FCTC). The WHO FCTC was adopted by the 56th World Health Assembly in May 2003 and became international law on February 27, 2005. It is the world's first public health treaty on tobacco control and encourages countries that ratify to develop and implement tobacco control policies, such as bans on direct and indirect tobacco advertising, tobacco tax and price increases, promoting of smoke-free public places and workplaces, and placement of health messages on tobacco packaging.The WHO FCTC, National Tobacco Control Action Plans, and the GYTS share the same goal: the development, implementation, and evaluation of effective tobacco control programs in all WHO member states. What the WHO FCTC and National Tobacco Control Action Plans ask countries to monitor, the GYTS can help to measure. The GYTS provides indicators for measuring achievement of several WHO FCTC articles .The synergy between the WHO FCTC, National Tobacco Control Action Plans, and GYTS offers countries a unique opportunity to develop, implement, and evaluate their comprehensive tobacco control programs. The reports included in this publication from Peru, Thailand, and Turkey are presented as examples of 'linking' GYTS data to the national tobacco control program efforts in each country.FCTC: Framework Convention on Tobacco Control; GTSS: Global Tobacco Surveillance System; GYTS: Global Youth Tobacco Survey; WHO: World Health Organization.The author declares that they have no competing interests."} +{"text": "Preventing Chronic Disease, CDC's online journal on public health policy, practice, and research (http://www.cdc.gov/pcd/). The Eriksen discussion focuses on whether the tobacco industry has forfeited its opportunity to participate in a traditional public-private partnership \u2014 the type that is sought by other industries \u2014 and if not, how it can partner successfully with the public health community. In the second half of his interview, Dr Eriksen considers the lessons learned from the tobacco control experience and how the public health community might work with the food and beverage industry to mitigate the obesity epidemic.Dr Michael Eriksen, former director of the Office on Smoking and Health at the Centers for Disease Control and Prevention (CDC) and director of the Institute of Public Health at Georgia State University, both in Atlanta, Georgia, was interviewed by Elizabeth Majestic with CDC's National Center for Chronic Disease Prevention and Health Promotion for The interview was filmed in November 2008.The tobacco industry has forfeited its opportunity to participate in traditional public-private partnerships, but does that mean that the public health community and the tobacco industry cannot work together to achieve a common objective of reducing the harm caused by tobacco use?Partnerships with the private sector should not be about money, but rather, should be considered when they support the achievement of public health objectives. The public health community also needs to hold the food and beverage industry accountable for its actions. For example, food and beverage companies should focus their efforts on both physical activity and good nutrition and they should evaluate efforts based on public health outcomes.Michael Eriksen has been director of the Institute of Public Health at Georgia State University since 2002. He received his undergraduate and graduate training at the Johns Hopkins University and has had a long and distinguished career in public health. Dr Eriksen has been employed in academia , the private sector (Pacific Telephone), state government , federal government (Centers for Disease Control and Prevention), and international organizations . He has published dozens of peer-reviewed articles on tobacco control, cancer prevention, and health promotion and is coauthor of The Tobacco Atlas with Judith Mackay. In 2004, the Georgia Cancer Coalition designated him as a Distinguished Cancer Scholar. Professor Eriksen teaches classes in the social and behavioral sciences, urban health, tobacco control, and global health. He is director of Georgia State University\u2019s Partnership for Urban Health Research."} +{"text": "The third AMIA Summit on Translational Bioinformatics built on the success of the 2008 and 2009 Summits. The Summit continues to highlight the multidisciplinary nature of this rapidly maturing research field and provides the opportunity to forge new transdisciplinary collaborations as the finest minds of the academia, industry, government and non-profit sector are brought together. The six tracks spanned the range from methods for the analyses of molecular through clinical measurements and informatics methods in genetics discoveries and clinical practice.1: Informatics Methods for the Integrative Analysis of Molecular and Clinical Measurements2: Computational Approaches to Finding Molecular Mechanisms and Therapies for Disease3: Informatics Concepts, Tools, and Techniques to Enable Integrative Translational Bioinformatics Research4: Relating and Representing Phenotypes and Disease for Translational Bioinformatics Research5: Informatics Methods Bridging Genetics Discoveries and Clinical Practice6: Dissecting Disease through the Study of Organisms, Evolution, and TaxonomyThe breadth and depth of the Summit continues to grow with twenty-five 90-minute scientific sessions. Original research was presented in forty-nine papers and abstract presentations covering all six tracks and spanning from methods to applications. Ten late breaking sessions covered work in press or published since the 2009 Summit including for example a presentation of the analysis of a full human genome in the clinical context. Six of the seven National Centers for Biomedical Computing presented on the relationship of their work to the themes of the Summit along with five other panels covering topics ranging from CTSAs to the eMERGE network. Four keynote presentations addressed the theme of paths toward genomic medicine. Spyro Mousses of TGen presented on \u201cUsing BioIntelligence to Search and Understand Individual Genomes\u201d illustrated the ideas in the context of individualized cancer therapy. Serge Saxonov of 23andMe presented on \u201cConsumers, Genomes and Research\u201d demonstrating how this approach could replicate findings from other studies and contribute to new genomic knowledge. Andrew Kasarskis of Sage Bionetwork presented on \u201cGetting from Heterogeneous Data to Actionable Shared Models of Biology\u201d and how these approaches can advance our fundamental understand of the underlying biology. Finally in what has become an annual tradition Russ Altman of Stanford presented a recap of some key developments and papers in his \u201cTranslational Bioinformatics Year in Review\u201d.http://summit2010.amia.org/session-details.Slides from selected tutorials, papers, and panels are publicly available at BMC Bioinformatics are extended and improved versions of the best papers accepted to the 2010 Summit on Translational Bioinformatics. These papers were nominated by members of the Scientific Program Committee and the Track Chairs and then subject to revision and additional peer review in collaboration with BMC Bioinformatics. These papers illustrate the span of the Summit and range from bioinformatics approaches to new knowledge discovery to informatics approaches to foster the application of this type of knowledge. In the following paragraphs we present an overview of the papers in this special issue and their organization.The eleven papers selected for this supplement to We start with three papers that describe original informatics methods for the integrative analysis of molecular and clinical measurements. The first paper deals with representing temporal processes in biology, while the other two deal with reasoning across molecular and clinical measurements. Chang and colleagues describeOne article focuses on computational approaches to finding molecular mechanisms and therapies for disease. More specifically, Zhang and collaborators investigThe next two articles discuss techniques to enable integrative translational research. The paper by Morgan et al. discuss The paper that follows is in the area of representing phenotypes and disease for translational bioinformatics research. The study by Lacson et al. assessesTwo papers focus on the important topic of informatics methods bridging genetics discoveries and clinical practice. Tatonetti and collaborators describeFinally, the last two papers present research work in the area of dissecting disease through the study of organisms, evolution, and taxonomy. The paper by Yang et al. focuses The authors declare that they have no competing interests.Both authors wrote and approved the manuscript. Mendon\u00e7a was a member of the Scientific Program Committee for the 2010 AMIA Summit on Translational Bioinformatics and the Guest Editor for this special issue; Tarczy-Hornoch was the Chair of the Scientific Program Committee for the 2010 AMIA Summit on Translational Bioinformatics.The authors and AMIA would like to dedicate this special issue to Marco Ramoni who played a vital role in the establishment of the AMIA Summit on Translational Bioinformatics having served as Track Chair for the 2008 and 2009 Summits and having served as a member of the Scientific Program Committee for the 2010 Summit. Prior to his sudden passing on June 8, 2010 Marco was one of the leading lights of the young field of translational bioinformatics. Marco was an Associate Professor of Pediatrics and Medicine at Harvard Medical School and research faculty at Children\u2019s Hospital, Boston and Brigham and Women\u2019s Hospital. Most recently he was the Director, Biomedical Cybernetics Laboratory, Harvard Medical School, Director, NLM Fellowship in Biomedical Informatics, Children's Hospital Informatics Program., and Associate Director of Bioinformatics, Harvard - Partners Center for Genetics and Genomics. He was also the Director of the course \u201cBiomedical Informatics\u201d at the Harvard-MIT Division of Health Sciences and Technology, core faculty of the course Genomic Medicine at Harvard Medical School and a member of the curriculum committee of the Cellular and Molecular Medicine track of the Medical Physics and Medical Engineering graduate program at Harvard-MIT Division of Health Sciences and Technology. He was co-founder of Bayesware LLC, a software company developing machine-learning programs based on Bayesian methods. He received a PhD in Biomedical Engineering and a BA in Philosophy (Epistemology) from the University of Pavia and his postdoctoral training from McGill University, Montreal (Canada). He held academic and visiting positions at the University of Massachusetts, the University of London (United Kingdom), the Knowledge Media Institute (United Kingdom), and the University of Geneva (Switzerland). He is author of well over 100 publications in genetics, biomedical informatics, statistics and artificial intelligence."} +{"text": "Twenty-six cases of Kaposi's sarcoma (KS) were recorded by the New South Wales Central Cancer Registry between 1972 and 1982, prior to the first AIDS diagnoses in Australia. The overall annual incidence was 0.47 per million. Incidence was three times higher in males. The highest incidence was in people born in the Middle East and in males born in southern and eastern Europe."} +{"text": "In early 1999, more than 160 senior physicians, public health officials, and nurses met to discuss London's tuberculosis (TB) control program. The program was examined against the public health response of New York City's Bureau of Tuberculosis Control during a 1988 to 1992 epidemic. This article outlines TB epidemiology and control in New York City 10 years ago and in London today to assess whether the kind of epidemic that occurred in New York could occur in London."} +{"text": "This is the second time I have come to England to the MRC Prion Unit. I have been working for the Kuru Research Project in the Papua New Guinea Institute of Medical Research for 6 years. Now, I am the Community Liaison Officer. I am also the community leader of Waisa Village. In our kuru studies, I work closely with Jerome Whitfield and Michael Alpers.In the early 1960s, when I first saw Michael Alpers in Waisa, I was 7 years old. Michael has always been a good friend of our family. The kuru research work went ahead successfully. Later Michael became the Director of the Papua New Guinea Institute of Medical Research. The Institute studies all the sicknesses of Papua New Guinea and we are pleased to have Prof. Peter Siba now as its Director.My late father Mr Puwa helped Michael with his work and supported him personally. He assisted with the examination of patients and the collection of samples from patients and others. He explained to the family and community why these samples of blood, brain and the like were needed. Working together, my father and Michael made a film on traditional salt making, a skill and technology that has now died out, since my father was the acknowledged salt maker of the village.Although most people still believe that kuru is caused by sorcery, there are a few of us who understand how it came to our people. I am very happy about all the research work for the last 50 years that has given us this understanding."} +{"text": "An automatic system for the determination of Zn, Cd, Pb and Cu by anodic stripping potentiometry using the oxygen dissolved in the sample as oxidant is reported. The system relies on the use of a PC-compatible computer for instrumental control and data acquisition and processing."} +{"text": "Trends in eye cancer mortality are presented for the USA and England and Wales during the period 1955-89. Mortality rates have fallen by 58% in the USA during this period. The fall in mortality is paralleled by an equal fall in incidence rates in the USA. In England and Wales, mortality rates and incidence rates have remained relatively constant during the last three decades. The explanation for these differences between the USA and England and Wales is unknown."} +{"text": "EHP was fortunate to have Steve Kleeberger and Matt Longnecker step up to serve with me as interim deputy editors in the absence of a full-time permanent editor. Steve and Matt have given generously of their time, expertise, and energy in helping to maintain EHP \u2019s scientific quality during a time of transition. Given EHP\u2019s broad scope of coverage, however, we felt the need to likewise broaden the scope of our science editorial expertise. Thus, we are pleased to welcome aboard two additional interim deputy editors.In January 2007, Toxicology and Applied Pharmacology since 2000 and serves on the editorial boards of Toxicology, the Journal of Toxicology and Environmental Health, and Toxicology Mechanisms and Methods.Michael P. Waalkes is a senior research toxicologist with the National Cancer Institute, where he serves as chief of the Inorganic Carcinogenesis Section, which is part of the Laboratory of Comparative Carcinogenesis stationed at the NIEHS campus in Research Triangle Park, North Carolina. Mike received his PhD in pharmacology and toxicology from West Virginia University. His postdoctoral studies at the University of Kansas School of Medicine focused on the cellular and molecular mechanisms of acquired tolerance to metal toxicity. His current research involves defining the mechanisms of action of the carcinogenic inorganics, including arsenic, lead, and cadmium. Mike is currently an adjunct professor of molecular toxicology at Duke University and an active member of the Society of Toxicology. He has been the editor of Epidemiology and has served on the editorial board of the American Journal of Epidemiology.Stephanie London is a senior investigator in the NIEHS Epidemiology Branch with a joint appointment in the Laboratory of Respiratory Biology. She received her MD and her DrPH degree in epidemiology from Harvard University. She completed a residency in internal medicine at the Massachusetts General Hospital and a residency in occupational and environmental medicine at Harvard, and is board certified in both fields. Stephanie was an assistant professor at the University of Southern California School of Medicine from 1989 through 1995, where she was part of a small team of investigators who founded a landmark study of health effects of air pollution in schoolchildren known as the Children\u2019s Health Study. She came to the NIEHS in 1995. Her work focuses on genetics and interactions between genetics, diet, and environmental pollutants in relation to asthma and chronic obstructive pulmonary disease. Stephanie is an editor for Journal of Exposure Science and Environmental Epidemiology, and is chair of the Environment and Reproduction Special Interest Group of the American Society for Reproductive Medicine.I would also like to take this opportunity to introduce our new medical editor, Russ Hauser, who succeeds Brian S. Schwartz. Russ is an associate professor of environmental and occupational epidemiology in the departments of Environmental Health and Epidemiology at the Harvard School of Public Health. He graduated from Clark University and the Albert Einstein College of Medicine, where he received his MD. He received his MPH and ScD from the Harvard School of Public Health, where he completed a residency in occupational medicine. He is board certified in occupational medicine. From 2000 to 2004, he served as deputy director of the Occupational and Environmental Medicine Residency, National Institute for Occupational Safety and Health Education and Research Center. Russ\u2019s research focuses on the effects of environmental and occupational chemicals on fertility and pregnancy outcomes. He is on the editorial board of the EHP \u2019s Grand Rounds and Environmental Medicine submissions, allowing the journal to advance in its mission of delineating relationships between the environment and human health. We are most grateful for his service to the journal.Finally, we bid a fond farewell to Brian, our outgoing medical editor. Brian served in this role from July 2004 through July 2007. During this time he significantly raised the bar for EHP. As always, we welcome your feedback.Stay tuned for news of a permanent editor-in-chief for"} +{"text": "Although we have excellent sources of information at the national level in the National Health Interview Survey and the National Health and Nutrition Examination Survey, and at the state and county levels in the Behavioral Risk Factor Surveillance System, each source has inherent limitations. More serious, however, are the issues raised by the 2002 report from the National Committee on Vital and Health Statistics (NCVHS) . That reThe Community Health Status Indicators (CHSI) project is a sigCommunity indicators.\u00a0The National Academy of Sciences and other organizations are involved in developing a set of key national indicators for the United States. These indicators would include health as one of their dimensions. Although this project is national, similar efforts are under way in U.S. communities, literally from coast to coast . The rapid growth of indicators or measures that inform the public and professionals alike in an easily accessible form is encouraging.Electronic health records. The use of electronic health records received a major boost from President George W. Bush and Secretary of Health and Human Services Michael Leavitt with establishment of the Office of the National Coordinator for Health Information Technology. Public health information is one of the foci of this organization and others, including the American Health Information Community (AHIC), a federally chartered advisory committee on the use and adoption of electronic health records. To assess the potential impact of electronic health records on health statistics and on public health decision making, the office of the Assistant Secretary for Health and the National Center for Health Statistics sponsored a workshop, and the report is available online (www.cdc.gov/nchs/data/nhcs/EMRworkshopsummaryjuly30.pdf).Healthy People program, now in its third decade. Although the Healthy People focus is national, it places a strong emphasis on developing data for local use. A recent paper shows that this goal is indeed feasible with a strong research base (Healthy People. Perhaps the leading force in developing new data sources and emphasizing the importance of standardization in definitions and methods is the Health and Human Services rch base .\u00a0Although these developments herald progress, critical gaps remain in our knowledge and data. As the nation grows more diverse in race, ethnicity, and various other factors, the need for adequate data to address health problems will multiply with each distinct population group. Furthermore, our definition of health must be broadened so that functioning is included as a part of our health assessment.Unfortunately, the current supporting resource base needs to be strengthened. CDC Director Julie Gerberding noted in CDC's Professional Judgment Budget Request that \"CDC's mission-critical health statistics and similar data systems are currently on life support. Investments have simply not kept pace with expenses and technological advances.\" As we work to achieve the promise of new sources and new tools, we also need to ensure that we do not damage the critical\u00a0national data infrastructure. Our investments in standards development must be balanced with investments in tools, training, analysis, and research.\u00a0Despite these constraints on resources, we continue to make progress in collecting and disseminating information crucial for public health decisions. The year 2007 marked the 50th anniversary of the National Health Interview Survey, an appropriate time to stress the gains we have made in health data and a commitment to working toward a future of relevant and high-quality information at every level of health decision making."} +{"text": "Felix D. Battistella, Professor and Chief of Trauma and Emergency Surgery at UC Davis Medical Center, died from cancer on January 22 at his home. He was 48.Dr. Battistella's unselfish dedication to his field, his colleagues and to the UC Davis Health System community was widely known.\" said Claire Pomeroy, Vice Chancellor for Human Health Sciences and Dean of the UC Davis School of Medicine. \"The many contributions he made have forever shaped the way we provide compassionate care for patients in their hour of need.\"\"magna cum laude. He received his medical degree from the UC Davis School of Medicine in 1985. For his postgraduate training, Battistella continued his association with UC Davis, completing an internship in the Department of Surgery in 1986, and finishing his residency in the department in 1990. He served as Chief Resident in the department from 1990 to 1991.Battistella figure was bornBattistella joined the Faculty of the Department of Surgery as an Assistant Professor in 1991. In addition to serving as Professor and Chief of Trauma and Emergency Surgery, Battistella was the Director of the department's residency program. He also served as the Chief of Staff from 2004 to 2006.In 1991, the senior class at the School of Medicine selected Battistella for its Outstanding Resident Teaching Award. That same year, he was voted Outstanding Chief Resident by his fellow residents in the Department of Surgery. In 1997, the Friends of Nursing at UC Davis Medical Center chose Battistella as its Physician of the Year.Dr. Battistella will be remembered most by any nurse who worked with him as the kindest, most compassionate surgeon. He was a highly skilled surgeon, but what set him apart was that he treated every one of his patients with compassion and caring. All of our nursing staff greatly admired Dr. Battistella.\"Carol Robinson, Senior Associate Director, Patient Care Services Administration, said, \"Dr. Battistella was a great teacher. In working with him and watching him with residents and students, you could see that he had a passion for teaching and mentoring. He also truly cared about the well-being of his patients.\"Cheryl Wraa, Manager of the UC Davis Trauma Program, said, \"Battistella is survived by his wife, Christine; daughters Claire and Mary; his mother, Amalia; and his sister, Nedra.World Journal of Emergency Surgery lost a great Editor and all emergency surgery science lost a great contributor.We think that"} +{"text": "A Funding Statement was incorrectly omitted from the article. The Funding Statement is: \"This work was supported by grant No. MH-3-7224 from the Chief Scientist\u2019s office of the Ministry of Health, Israel to AA and by Grant No. 2009179 from the United States-Israel Bi-national Science Foundation (BSF) to TH and AA. We are also thankful for the support from the Jees & Midred Fisher Family\u2019s Cardiology Research Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There are errors in the Funding statement. The correct Funding statement is as follows: Researcher Marcus Sheaves was supported by the Marine Biodiversity Hub through funding from the Australian Government's National Environmental Science Program. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In the Model for analyzing individual events subsection of the Methods, there are errors in third and fourth equation of Model (2). Please view the complete, correct equations here:The publisher apologizes for the error."} +{"text": "The Funding section for this paper is incorrect. The publisher apologizes for the error. The correct funding information is as follows: This work was supported by the Grant for Specially Promoted Research from Kanazawa Medical University , the Japanese Ministry of Education, Culture, Sports, Science and Technology (MEXT)-Supported. Program for the Strategic Research Foundation at Private Universities , and the 4th Community Medicine Promotion Grant of the Sugiura Memorial Foundation (Dr. Yukihisa Matsuda). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The authors of \u201cUsing Behavioral Intervention Technologies to Help Low-Income and Latino Smokers Quit: Protocol of a Randomized Controlled Trial\u201d :e127) would like to change the Acknowledgements section of their paper to the following:\u00a0\u201cThis project was partially supported by funds provided by The Regents of the University of California, Tobacco-Related Diseases Research Program, Grant Number No. 24RT-0027. The opinions, findings, and conclusions herein are those of the authors and not necessarily represent those of The Regents of the University of California, or any of its programs.\u00a0Programming and development of the web app for this project is being carried out by the Center for Behavioral Intervention Technologies at Northwestern University Feinberg School of Medicine.\u201dThe originally published acknowledgement has only the first sentence. This correction has been made in the online version of the paper on the JMIR Research Protocols website on September 23, 2016, together with publishing this corrigendum.A correction notice has been sent to PubMed, and the publication was resubmitted to Pubmed Central and other full-text repositories."} +{"text": "The following information is missing from the Funding section: The National Institute of Health, R01 MH079406 (RES) and New Jersey Health Foundation (RES). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The publisher apologizes for the error."} +{"text": "In the original article, we neglected to include the funder Research Grants Council of Hong Kong, General Research Fund (GRF/HKU/17609417), to Temmy Lee Ting Lo and Rainbow Tin Hung Ho.Due to this omission, the following Funding statement will be added to the original article:This work was supported by the General Research Fund, Research Grants Council of Hong Kong .The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The authors of \u201cHow Online Communities of People With Long-Term Conditions Function and Evolve: Network Analysis of the Structure and Dynamics of the Asthma UK and British Lung Foundation Online Communities\u201d :e238) wish to add the following sentence to the Acknowledgments:The correction will appear in the online version of the paper on the JMIR website on September 4, 2018, together with the publication of this correction notice. Because this was made after submission to PubMed, Pubmed Central, and other full-text repositories, the corrected article also has been re-submitted to those repositories."} +{"text": "Scientific Reports 10.1038/s41598-018-21140-3, published online 09 February 2018Correction to: In the original version of this Article, Kuan-Yi Wu and Ing-Tsung Hsiao were incorrectly listed as equally contributing authors.In addition, the Acknowledgements section in this Article was incomplete.\u201cWe thank Avid Radiopharmaceuticals Inc. for providing the precursor for the preparation of 18F-florbetapir. This study was carried out with financial support from the National Science Council and the Ministry of Science and Technology, Taiwan , in addition to grants from the Research Fund of Chang Gung Memorial Hospital and the Center for Advanced Molecular Imaging and Translation.\u201dnow reads:\u201cWe thank Avid Radiopharmaceuticals Inc. for providing the precursor for the preparation of 18F-florbetapir. This study was carried out with financial support from the National Science Council and the Ministry of Science and Technology, Taiwan , in addition to grants from the Research Fund of Chang Gung Memorial Hospital and the Center for Advanced Molecular Imaging and Translation. This study was also supported by the Ministry of Health and Welfare, Taiwan .\u201dThese errors have now been corrected in the PDF and HTML versions of the paper, and in the accompanying Electronic Supplementary Material."} +{"text": "The following information is missing from the Funding statement: This article was funded by the Qatar National Library. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Following publication of the original article , the autThe acknowledgments currently read:We would like to thank the Simmons Center for Cancer Research for the financial support, Dr. Juan Arroyo (BYU) for his assistance in tissue staining and confocal microscopy, Dr. Himelda Chavez , for her pathology expertise and help in IHC analysis, and a team of surgeons at Utah Valley Regional Medical Center for providing normal and malignant colon tissue.The corrected acknowledgements would read:http://cancergenome.nih.gov).We would like to thank the Simmons Center for Cancer Research for the financial support, Dr. Juan Arroyo (BYU) for his assistance in tissue staining and confocal microscopy, Dr. Himelda Chavez , for her pathology expertise and help in IHC analysis, and a team of surgeons at Utah Valley Regional Medical Center for providing normal and malignant colon tissue. Results from this study are in part based upon data generated by The Cancer Genome Atlas and managed by the United States National Cancer Institute and National Human Genome Research Institute (see"} +{"text": "In the original publication, the funding information was incorrectly published. The correct funding information is provided in this correction.This work is supported by grants from the Projects of International Cooperation and Exchanges Ministry of Science and Technology of China (2013DFG32390) and the National Natural Science Foundation of China (31472059) to X.S. X.S is a recipient of the Young Thousand Talents program (KJ2070000026)."} +{"text": "Hadley Weiss is not included in the author byline. Hadley Weiss should be listed as the 25th author and affiliated with LS50: Integrated Science Freshman Class, Harvard University, Cambridge, Massachusetts, United States of America. The contributions of this author are as follows: Investigation and Writing\u2013Review & Editing.The publisher apologizes for the error."} +{"text": "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There is an error in the Funding statement. The publisher apologizes for the error. The correct Funding statement is: This research was supported by the National Natural Science Foundation of China (URL:"} +{"text": "There is an error in the Funding statement. The correct Funding statement is as follows: This research was supported by grants from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI) funded by the Ministry of Health & Welfare, Republic of Korea . The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Sudre, and Sebastien Ourselin. These authors\u2019 affiliations are now correctly noted as School of Biomedical Engineering & Imaging Sciences, King\u2019s College London, London, UK. Additionally, a statement from the Acknowledgments section was omitted. The Acknowledgments have been updated to include the following: \u201cWe are grateful to the MS Society for its support of the Multiple Sclerosis Research Centre at the Queen Square Institute of Neurology where this work was completed.\u201d The HTML and PDF versions of the Article have been corrected."} +{"text": "Polymers,Dear readers of Polymers Young Investigator Award to:Finally, after an extensive voting period, we are proud to present the first winner of the Polymers Investigator Award from 38 candidates who were proposed by at least two colleagues in their field of expertise. Fifteen of the candidates are working in the United States, 13 in Europe and 10 at universities in Asian countries. The proposed candidates represented a diverse range of fields in polymer science. Dr. Campos received his Ph.D. with Professor Miguel A. Garcia-Garibay and Professor Kendall N. Houk at University of California, Los Angeles (UCLA) in 2016 and did his postdoc training at the University of California, Santa Barbara (UCSB) under the supervision of Craig J. Hawker. At the age of 37, Dr. Campos has already achieved an extraordinary standing in the polymers community. His excellent work focuses on the design and application of polymeric materials, for example, solar cells and organic light emitting diodes; all topics of high societal and economic impact. His research has been featured in highly ranked journals such as the Nature family, Angewandte Chemie, and Journal of the American Chemical Society (JACS), to name a few. To date, he has co-authored over 60 articles and has received numerous awards, including the American Chemical Society (ACS) Arthur C. Cope Scholar Award, The Office of Naval Research (ONR) Young Investigator Award, The National Science Foundation (NSF) CAREER Award, 3M Non-Tenured Faculty Award, Cottrell Scholar Award, The Inter-American Photochemical Society (I-APS) Young Faculty Award, the Journal of Physical Organic Chemistry Award for Early Excellence. This finally led to invitations to numerous prominent lectures all over the world. Moreover, his work constitutes not only high level basic research, but has proven to be highly relevant in industry which is reflected in 10 patents filed by him and his coworkers.who is an assistant professor at the Chemistry Department of Columbia University, USA. He was selected by the evaluation committee of Polymers conference.In addition to the cash prize and plaque, Dr. Campos will be an invited speaker at the 2018 Polymers Editorial office staff and editorial board members, I wish to congratulate Dr. Campos on his excellent performance and wish him all the best for his future career.On behalf of the"} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This study is part of the research project \u201cResistance Training in Youth Athletes\u201d that was funded by the German Federal Institute of Sport Science (ZMVI1-08190114-18). In addition, we acknowledge the support of the Deutsche Forschungsgemeinschaft (DFG) and Open Access Publishing Fund of University of Potsdam, Germany. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The following information is missing from the Competing Interests statement: WHO-IARC: Where authors are identified as personnel of the International Agency for Research on Cancer / World Health Organization, the authors alone are responsible for the views expressed in this article and they do not necessarily represent the decisions, policy or views of the International Agency for Research on Cancer / World Health Organization.Norwegian Immunization Registry SYSVAK provided data, but is not responsible for the analyses and interpretation in this article.The publisher apologizes for the error."} +{"text": "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There are errors in the Funding section. The correct funding information is as follows: This work was supported by a grant of the Deutsche Forschungsgemeinschaft to DJ (DFG Je 152/17-1) and Vikas Sharma is a recipient of a research fellowship from the Ministry of Human Resource Development (MHRD), India ("} +{"text": "S2 Text in the original article to disclose the role of the funder Illumina in this study, the financial disclosure was not updated to reflect the same information. The Funding should read:Following the inclusion of S2 Text. The other funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.REC, CMD, TCR, and AM had funding from the Department for International Development from the United Kingdom award (204074\u2013101) and Department of Foreign Affairs and Trade Australian Government award (70957). TCR had funding from National Institute of Allergy and Infectious Diseases (grant number: R21AI135756). MS was supported from the National Heart, Lung, and Blood Institute at the National Institutes of Health (grant number: T32HL134632). Illumina supplied early-access iSeq100 sequencing reagents; some experiments were conducted at Illumina by Illumina employees; the role of the funder Illumina is further described in"} +{"text": "This article has been retracted: From the The First Hospital of China Medical University Professor Committee, in response to the Journal\u2019s request:The First Hospital of China Medical University Professor Committee convened the 14th plenary meeting to investigate Yu-Ji Li\u2019s paper published in Oncotarget , in response to the Journal\u2019s editorial board\u2019s request. A thorough investigation and open discussion was held in the meeting that included all department members in charge of scientific affairs.At the meeting, Dr. Yu-Ji Li reported all events in the production of his paper. He admitted the problems in the publication that were identified by the Journal and acknowledged that he was responsible for the incidents. The chairman of Dr. Li\u2019s department also detailed on the incidents and pointed out the loopholes in the paper. Inquiries were made and documents were carefully checked by the Committee members. The Professor Committee agreed that Yu-Ji Li was negligent in his research attitude and was unable to offer complete original materials. After the discussion, the decision was made as the following:1) Due to repeated and manipulated use of figures that misled the scientific conclusion, we ask for retraction of the paper by Dr. Yu-Ji Li from the Journal. (Dr. Li himself previously attempted to retract but his plea was unclear in expression).2) Apologies must be issued to the Journal and its readerships for the authors\u2019 negligence and lack of rigorous academic integrity.3) The University will take strict measures against the misconduct by the authors of the paper according to relevant institutional regulations.https://doi.org/10.18632/oncotarget.8289Original article: Oncotarget. 2016; 7:31993-32005."} +{"text": "In the original article, there was an error in the funding section.Funding section and now states:A correction has been made to the \u201cThis work was funded by the EPFL Blue Brain Project Fund and the ETH Board Funding to the Blue Brain Project. The work has been performed as an in-kind contribution to the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement no. 604102 (Human Brain Project) and the European Union's Horizon 2020 Framework Programme for Research and Innovation under Grant Agreement No. 720270 (Human Brain Project SGA1).\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The author Mabel Kwong was inadvertently omitted from the list of authors during the production stage of the paper \"Internet Protocol Television for Personalized Home-Based Health Information: Design-Based Research on a Diabetes Education System\" :e13). The author Mabel Kwong should have been added after Ken Clarke in the original published manuscript. This error has been corrected in the online version of the paper on the JMIR Research Protocols website on March 18, 2014, together with publishing this correction notice. This was done before submission to Pubmed Central and other full-text repositories."} +{"text": "Qualifying researchers may apply to access the data by contacting Office of Statewide Health Planning and Development (OSHPD) of California, Information Services Division (dataandreports@oshpd.ca.gov).The Data Availability statement for this paper is incorrect. The correct statement is: There are ethical and legal restrictions on sharing the data set due to patient privacy. Therefore, legal restrictions related to agreements made with the Office of Statewide Health Planning and Development (OSHPD) of California ("} +{"text": "There are errors in the Funding statement. The publisher apologizes for the errors. The correct Funding statement is as follows: This study was supported by the Fundamental Research Funds for the Central Universities of Central South University (1053320182836). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The following information is missing from the Funding section: This article was funded by the Qatar National Library. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In the original article, we neglected to include the funder \u201cNational Natural Science Foundation of China, 71872102.\u201d The updated funding statement is mentioned below:This research was supported by National Natural Science Foundation of China (No. 71872102).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "There is an error in the Funding statement. The correct number for the Foundation for Science and Technology I&D 472 is (UID/DTP/00447/2019). The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "In the original article, we incorrectly used the university internal grant code which should be replaced by the Chinese Medicine Research Fund in University of Oxford. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.The funding statement should read:This work was supported by the Chinese Medicine Research Fund in University of Oxford. The grant was sponsored by Shaanxi Momentum Pharmaceutical Co., Ltd.Shaanxi Momentum Pharmaceutical Co., Ltd. had no involvement in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. All authors declare no conflict of interest."} +{"text": "The following information is missing from the Funding statement: Publication of this article was funded in part by the Kansas State University Open Access Publishing Fund.In the Methods section, there is a typographical error in equation 3. Please view the complete, correct equation here:"} +{"text": "The following information is missing from the Funding section: This work was partly funded by the Marine Biodiversity Hub, a collaborative partnership supported through funding from the Australian Government's National Environmental Science Programme. The funding body was not involved in the process of study design, data collection and analysis, decision to publish, or preparation of the manuscript.There is an error in the Acknowledgments section. The name \"Ashley Leeman\" is spelled incorrectly. The correct spelling is: Ashley Leedman."} +{"text": "The authors of the published article \u201cInternet Search and Krokodil in the Russian Federation: An Infoveillance Study\u201d wish to add additional information to the \u201cAcknowledgments\u201d section, so it now reads as follows:The work of Peter Meylakhs was funded by the Basic Research Program of the National Research University Higher School of Economics, Russia.\u00a0The authors would also like to thank Ms Svetlana Chernova in Ukraine for her assistance with data collection and coding, Associate Professor James Gillespie at the Menzies Centre for Health Policy, University of Sydney for his advice and support, and Ms Anya Sarang from the Andrey Rylkov Foundation in Moscow for facilitating contacts within Russia.The first sentence was unintentionally omitted in the original submission. The publication on the JMIR site was amended accordingly, and a new version was submitted to PubMed Central."} +{"text": "On September 25 2017, Jay A. Siegel, PhD died at his home in Mt. Arlington, New Jersey at the age of 71. At the time of his death due to heart failure, Dr Siegel was highly regarded internationally as a forensic chemist, educator and prominent advocate for quality in the forensic sciences. Jay was born in Washington, DC, USA on April 16 1946. He received a PhD degree in chemistry from The George Washington University in Washington, DC, USA in 1975. Early in his career, Jay developed strong interest in the forensic applications of chemistry. Following employment as a forensic chemist with the Virginia Bureau of Forensic Sciences, Jay launched decades of professional activity as an educator and research scientist with Metropolitan State College in Denver, Colorado, Michigan State University in East Lansing, Michigan and Indiana University-Purdue University in Indianapolis, IN, USA. His university experience included serving as department chair but primarily focused on educating many undergraduate and graduate students who have in their own subsequent careers made many contributions to forensic science. His international recognition included visiting professor roles at universities in Australia and China.Journal of Forensic Sciences from 1983 to 2008 and the Forensic Sciences Research from 2016 to 2017 and as a commissioner for the Forensic Science Education Programs Accreditation Commission (FEPAC) for six years beginning in 2003.Jay received many prestigious honours and awards during his long career. These include the Paul L. Kirk Award in 1975 from the Criminalistics Section of the American Academy of Forensic Sciences (AAFS). Jay was named Distinguished Fellow by the AAFS in 2009. Also in 2009, he received the Distinguished Alumni Award from The George Washington University and in 2012 the Distinguished Service Award from the Midwest Association of Forensic Scientists. Jay served on the editorial boards of the Throughout his career, Jay was a strong advocate for quality and scientific rigour in the practice of forensic science. He was selected to serve on the National Research Council Committee that led to the 2009 publication of the influential \u201cStrengthening Forensic Science in the United States: A Path Forward.\u201d Jay authored books and numerous publications in relevant scientific journals within his interests in forensic chemistry and the forensic sciences in general. He was an editor (with Pekka J. Saukko) for the Encyclopedia of Forensic Sciences published in 2012 by Academic Press.Journal of Forensic Sciences. During this time, I recall many conversations with Jay concerning the quality of publications in the forensic sciences, especially relating to his own specialty of forensic chemistry. In my additional role as Chair of the Forensic Science in Focus Book Series for AAFS, I called Jay, challenging him to organize a quality, edited book on forensic chemistry to be published in the series that would address many of the shortcomings we had discussed previously. Jay accepted this challenge and responded with a carefully edited manuscript with contributions from many of his colleagues. The volume \u201cForensic Chemistry: Fundamentals and Applications\u201d was published by Wiley-Blackwell in 2015.Over the years, I enjoyed many discussions with Jay regarding issues within the forensic sciences and developments relating to AAFS. He could always be counted upon for strong, thoughtful positions on topics of mutual interest. From 2008 to 2016, I served as the Book Review Editor for the In my view, Jay Siegel represents an ideal role model for all of us in the forensic sciences. His work demonstrated how academia and forensic practice are not mutually exclusive. Like Jay, we should all strive for the highest quality in forensic science possible and embrace the challenges of constructive criticism and the advances of new research."} +{"text": "In the Funding section, the grant number from the National Science Foundation, China is listed incorrectly. The correct grant number is: 71801089."} +{"text": "The Maxwell A. Pollack Award for Productive Aging recognizes instances of practice informed by research and analysis, research that directly improved policy or practice, and distinction in bridging the worlds of research and practice. The award lecture will be presented by the 2018 recipient, Karen Fredriksen-Goldsen, PhD, University of Washington. This award is generously funded by The New York Community Trust."} +{"text": "In the original article, we neglected to include the funders. HA-K acknowledges the Higher Education Committee for Education and Development (HCED), Office of Prime Minister, Iraq for financial support. AA and HA acknowledge the Royal Embassy of Saudi Arabia \u2013 Cultural Bureau (UK) for financial support. RE-G acknowledges WELMEC, a Centre of Excellence in Medical Engineering funded by the Wellcome Trust and EPSRC, under grant number WT 088908/Z/09/Z for financial support.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way.The original article has been updated.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The heights by great men reached and kept were not attained by sudden flight, but they, while their companions slept, were toiling upward in the night. (Henry Wadsworth Longfellow)Prof. Samuel Fehrsen see and 2 waHe was the first head of the Department of Family Medicine at the former Medical University of Southern Africa \u2013 MEDUNSA from 1977 to 1996, spanning a period of 20 years. During his tenure at MEDUNSA, Prof. Sam Fehrsen established a family medicine department that trained and produced many family physicians in South Africa, Southern Africa and Central Africa. In 1995, through his visionary leadership, in collaboration with the Evangelical Church of the Congo (ECC), the very first and only Family Medicine postgraduate training programme in the Democratic Republic of the Congo was established. Some of our collaborators from the Democratice Republic of the Congo (DRC) include Prof. Leon Kintaudi and Dr Philippe Lukanu \u2013 both based at the Protestant University of the Congo (UPC). The DRC programme has matured to the extent that the department continues to support the local training of family medicine registrars through the establishment of a Memorandum of Understanding with the UPC in 2011.In addition, he played a pivotal role in the establishment of the first Family Medicine programme in Kenya at the Moi University, Eldoret. Most of the first cohort of family physicians who graduated from that university are now heads of departments (HODs) (Family Medicine) in at least three universities in Kenya. In southern Africa, close to 200 family physicians from Swaziland, Namibia, Lesotho, Zambia and Botswana trained under his leadership through the vocational Family Medicine postgraduate programme at MEDUNSA. A number of family physicians have further emigrated to Canada, Australia and New Zealand. The current HOD of Family Medicine at the University of Namibia (Dr Felicia Christians) is an alumnus of University of Cape Town.SA Family Practice Journal as well as a role model, mentor and friend to many. Late Prof. Sam Fehrsen has left footprints on the sands of time that we will never forget. In his quiet way, he succeeded in establishing and spreading the specialty of Family Medicine in South Africa, southern Africa and Africa. May his gentle soul rest in peace!In South Africa, the Family Medicine programme that the late Prof. Sam Fehrsen ran for 20 years produced many prominent family physicians. The current HODs of Family Medicine at Walter Sisulu University (WSU), University of KwaZulu-Natal (UKZN), University of Limpopo (UL), University of the Witwatersrand (Wits) and my humble self are all his mentees and products. He was awarded an honorary degree \u2013 Doctor of Medicine (MD) \u2013 by the WSU in 2009 for his tremendous contributions to rural health services and training in the former Transkei. What an achievement for someone who was always soft-spoken, affirmative, disciplinarian and a missionary at heart. His command of the isiXhosa language amazed us all the time from his earlier medical professional life at Mt Ayliff Hospital, Eastern Cape. He was a pioneer of the South African Academy of Family Practitioners that later transformed to the South African Academy of Family Physicians, a past president of the Academy, past editor of the"} +{"text": "In \u201cPredictors of Patients\u2019 Loyalty Toward Doctors on Web-Based Health Communities: Cross-Sectional Study\u201d by Wu et al :e14484), a minor error in the typesetting stage of publication resulted in the Acknowledgments section not being included in the final version of the article.The Acknowledgments section has been added to the paper and appears as follows:This work was partially supported by grants from the National Natural Science Foundation of China .The correction will appear in the online version of the paper on the JMIR website on November 7, 2019, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "The author wishes to make the following corrections to this paper . Due to with:Moreover, in the published article , the autFunding: This research was funded by the Faculty of Biotechnology and Food Science, Wroc\u0142aw University of Environmental and Life Sciences, grant number B030/0003/1 and under the program of the Minister of Science and Higher Education \u201cStrategy of Excellence University of Research\u201d in 2018\u20132019 project number 0019/SDU/2018/18 in the amount of PLN 700 000.The authors would like to apologize for any inconvenience caused to the readers by the change. The change does not affect the scientific results. The manuscript will be updated, and the original will remain online on the article webpage, with a reference to this Correction."} +{"text": "The original version of this article unfortunately contained a mistake. Information was missing in the acknowledgements section. The correct information is given below.Acknowledgements We thank the Mountain Rescue South Tyrol (Bergrettungsdienst im Alpenverein S\u00fcdtirol) for putting their indoor climbing facility at our disposal. Furthermore, we thank the companies Mindray Bio-Medical Electronics Co., Ltd., Masimo Corporation, Akern and Salewa for lending the equipment. We would like to acknowledge the support of Prof. Luciano Bernardi in interpreting the data on baroreflex sensitivity. The authors thank the Department of Innovation, Research and University of the Autonomous Province of Bozen/Bolzano for covering the Open Access publication costs."} +{"text": "The affiliation for the second author is incorrect; the correct affiliation is not indicated. Liam Bailey is not affiliated with #2, but with: Department of Evolutionary Genetics, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany."} +{"text": "The M. Powell Lawton Award is presented annually to an individual who has made outstanding contributions from applied research that has benefited older people and their care. The lecture will be given by the 2018 recipient, Carol Whitlatch, PhD, Benjamin Rose Institute on Aging. The session will also include the presentation of the 2019 Lawton Award. The 2019 Lawton Award recipient is Barbara Resnick, PhD, CRNP, FGSA, of the University of Maryland. Supported by the Polisher Research Institute of the Madlyn and Leonard Abramson Center for Jewish Life."} +{"text": "Perl\u201d was not included as an author in the published article and has now been added. All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.Acknowledgments section:Additionally, due to the addition of the author Scott M. Perl, a correction has been made to the \u201cA portion of this research took place at the Jet Propulsion Laboratory, California Institute of Technology, under a contract with the National Aeronautics and Space Administration. General ideas presented here developed out of various NASA and NSF grants to the authors, along with partial support from the NASA Astrobiology Institute (project: Rock Powered Life) to JS, NSF Coupled Natural Human Systems (Award #1617473) to BB, and NSF National Robotics Initiative (Award #IIS-1526667) for DT. We thank the reviewers for input that improved this manuscript.\u201dThe authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Addendum to: npj Schizophrenia 10.1038/npjschz.2015.16, Published online 6 May 2015In the original published version of the manuscript, no data availability statement was included. This addendum has been added to include the following data availability statement \u201cData obtained from the patients in the Taiwan cohort were used with agreement from National Taiwan University Hospital. Data acquisition was funded by grant NSC99-3112-B-002-030 and was collected at National Taiwan University Hospital around 2010. Data obtained from the patients in the Huaxi cohort were used with agreement from Sichuan University. Data acquisition was funded by National Nature Science Foundation of China (81130024), National Key Technology R & D Program of the Ministry of Science and Technology of China during the 12th Five-Year Plan (2012BAI01B06). Reasonable requests for access to the data should be directed to Tao Li, The Mental Health Center and the Psychiatric Laboratory, West China Hospital, Sichuan University. Data obtained from the patients in the Xiangya cohort were used with agreement from the Second Xiangya Hospital of Central South University in China. Reasonable requests for access to the data should be directed to Zening Liu, Institute of Mental Health, Second Xiangya Hospital, Central South University. Data obtained from the patients in the Nottingham cohort were used with agreement from Nottingham University. Data acquisition was funded by MRC (grant reference G0601442). Reasonable requests for access to the data should be directed to Peter F Liddle, Division of Psychiatry and Applied Psychology, University of Nottingham.\u201d"} +{"text": "In the publication of this article , there iThe error: In the initially published version of this article, the caption for Figs. The correct captions for Figs."} +{"text": "There is an error in the Funding statement for GY. GY was supported by grants MH107367 and HD085902 from the National Institutes of Health and the Seed Grant BRFSG-2014-14 from the Brain Research Foundation.Funding:As a result, a correction has been made to the \u201cThis work was supported by the Netherlands Organisation for Scientific Research (NWO) Rubicon Fellowship (019.163LW.032) (to MvdH); the Brain Foundation, the Walker Family, and the Perpetual Impact Philanthropy (grant IPAP2017/0717) (to CB); the G. Harold & Leila Y. Mathers Charitable Foundation, JPB Foundation, and the NIH (to FG). GY was supported by MH107367 and HD085902 from the National Institutes of Health and the Seed Grant BRFSG-2014-14 from the Brain Research Foundation.\u201dAdditionally, in the original article, there is an error. The conflict of interest statement is incomplete.Conflict of interest statement:A correction has been made to the \u201cGY is co-founder, member of the Board of Directors, on SAB, equity holder, and paid consultant for Locana. The terms of this arrangement have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.The authors apologize for these errors and state that they do not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Correction to: Mol Autism (2019) 10:29https://doi.org/10.1186/s13229-019-0280-6Following publication of the original article , we haveAll EEG studies and analyses were performed with institutional review board (IRB) and/or National Research Ethics Service approval.The Ethics statement should read:The EEG data collected for subject 801\u2013015 was collected at University of Tennessee Health Science Centre (UTHSC) under an IRB approved protocol with informed consent, the transfer of this EEG data to University of California, Los Angeles (UCLA) for analysis was not approved by the UTHSC IRB. The data transfer was approved by the UCLA IRB with the understanding that the data file was de-identified, however, patient identifiers were included on the EEG disk in error. The EEG data for this subject are now stored on the UCLA server fully de-identified. All other subjects were collected at UCLA under an IRB approved protocol. The data and results presented in this article remain unaffected, and this correction has been published at the request of the Research Integrity Officer at UTHSC.The authors gratefully acknowledge the support of the Economic & Social Research Council . Prof Mark Hawley is a Theme Lead for the National Institute for Health Research (NIHR) Devices for Dignity MedTech Co-operative (MIC). The work of the Devices for Dignity MIC is funded by the NIHR. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care."} +{"text": "The Swedish Medical Products Agency (MPA) is the Swedish national authority responsible for regulation and surveillance of the development, manufacturing and marketing of medicinal products. Relenza, a neuraminidase inhibitor used in the treatment of influenza, has been evaluated by both the Swedish MPA and the United States Food and Drug Administration.In a paper by Mulinari et al. , the div"} +{"text": "Subscribe to the newsletters and see the WONCA Africa page there at https://www.wonca.net/AboutWonca/Regions/Africa.aspx.As the new president of the World Organization of National Colleges, Academies and Academic Associations of General Practitioners/Family Physicians (WONCA) Africa, I realise that many family doctors in Africa know very little about WONCA. I hope that this editorial and other communications will begin to ensure every family doctor in Africa knows and supports WONCA. WONCA was set up in 1972. It built up a great brand value over the years. This was retained in developing a short name for the organisation: WONCA, the World Organisation of Family Doctors. WONCA Council is made up of 118 member organisations from colleges, academies and associations of family doctors in 131 countries across the world. The mission of WONCA is to improve the quality of life of people of the world through defining and promoting its values, including respect for universal human rights, including gender equity, and by fostering high standards of care in general practice/family medicine. It represents and acts as an advocate for its members at an international level where it interacts with world bodies such as the World Health Organization (WHO). A small WONCA Executive team meets regularly between biannual WONCA Council meetings. The 13 members of WONCA Executive team are the world president, president-elect, immediate past president, regional presidents from each of the seven WONCA regions \u2013 Africa, Asia Pacific, East Mediterranean, Europe, Iberoamericana-CIMF, North America and South Asia \u2013 and three members-at-large. The WONCA World Secretariat is currently located in Bangkok. WONCA has rich resources on its website, WONCA Africa Region is made up of ten paid-up member organisations as of October 2018: Association of Family Physicians of Uganda, Association of General and Private Medical Practitioners of Nigeria, College of Primary Care Physicians of Zimbabwe, Faculty of Family Medicine in the National Postgraduate Medical College of Nigeria, Kenya Association of Family Physicians, Lesotho Medical Association, Society of Family Physicians of Nigeria, South African Academy of Family Physicians, Society of Family Physicians of Ghana and the Faculty of Family Medicine (Ghana Chapter) in the West African College of Physicians. Africa was right there in WONCA in 1972. Recent presidents included Bruce Sparks, Khaya Mfenyana, Sylvester Osinowo, Matie Obazee and Henry Lawson. Africa has been a very difficult terrain for growing WONCA and is recognised as such by WONCA. WONCA had a memorable world onference in South Africa in 2001 and elected Bruce Sparks as its world president for 2004\u20132007. The first WONCA Africa conference was held in Nigeria in 2000, with a second conference held after many years in South Africa in 2009. This continued with Zimbabwe (2012), Ghana (2015) and South Africa (2017). Much of the recent success in holding conferences has come from Belgian support for African family medicine, especially the Primafamed Network and its collaboration with WONCA conferences.I was elected as president-elect at the WONCA Africa Council Meeting before the WONCA Conference in Rio de Janeiro, Brazil (2016) to serve under President Henry Lawson (2016\u20132018) and to then serve as President from 2018 to 2020. WONCA Africa Council met again before the WONCA Conference, Seoul, South Korea (October 2018) to elect the new WONCA Africa Executive Committee see : Dr Dan www.WoncaAfrica2019.com. Register and come to Uganda to interact with an array of international speakers on the theme \u2018People-centred PHC\u2019. The year 2019 will be a momentous year, with the implementation of the Astana Declaration becoming a key issue amongst governments. We intend to engage with the WHO globally and in Africa to facilitate relationships between ministries of health and WONCA member organisations in Africa. We will be inviting the WHO Regional Director to speak at our Regional Conference and to meet leaders of our member organisations. There is a number of other plans: building membership of WONCA Africa, especially academic departments of family medicine; building resources for academic departments including research collaborations; improving women and young doctor participation in WONCA Africa and member organisations; strengthening African involvement in WONCA working parties and special interest groups; improving funding of WONCA Africa; adopting regional bylaws to ensure good governance of WONCA Africa; and improving communications with ordinary family doctors as well as member organisations.There are many plans but the first priority is to ensure that the next WONCA Africa Conference in Kampala, Uganda, 06\u201308 June 2019, is a success. The website is available at https://www.woncaafrica.org, and @WoncaAfrica on Facebook, Twitter and Telegram to keep abreast of things at WONCA Africa.This is the first editorial from WONCA Africa. There will be more details on these plans as we proceed. Check the WONCA Africa website at"} +{"text": "The authors wish to make the following corrections to their paper :The middle initial has been added to the name of co-author Sanghamitra Misra. The correct name is Sanghamitra M. Misra.https://imconsortium.org/about/introduction/ (accessed on 27 August 2018).Academic Consortium for Integrative Medicine and Health. Available Online: The citation website link to the first reference in the original paper is now unavailable. The correct reference is as follows:In addition, we found that the organization name listed in Section 4 (p. 5) and in the acknowledgments section is incorrect\u2014the reference to the Academic Consortium for Integrative Medicine and Health (ACIHM) is incorrect. The correct organization name is the Academic Collaborative for Integrative Health (ACIH). The corrected last sentence of the second paragraph in Section 4 is as follows:Specifically, with organizational support by John Weeks, a pioneer in collaborative interprofessional IM efforts over several decades, representatives from the Academic Collaborative for Integrative Health (ACIH) participated in a comprehensive pre-summit survey.The corrected acknowledgments are as follows:Acknowledgments: The Pediatric Integrative Medicine Leadership Initiative would like foremost to thank the Marino Health Foundation for its generous support of the 2015 and 2016 summits and related work. The PIMLI would also like to acknowledge the American Academy of Pediatrics, in particular Teri Salus and Anne Gramiak, for their logistical support coordinating and facilitating the 2015 PIMLI Summit. PIMLI leaders are extremely grateful to John Weeks and the ACIH committee who helped in preparation for the 2015 summit, and to Kiwi Magazine and the Moms Meet Network for assistance with the 2015 Parent Survey. Finally, the PIMLI group thanks the members of the PIM community who participated in the 2015 and 2016 professional surveys. Thank you to all the PIMLI leaders involved, in addition to the authors: Michelle Bailey, David Becker, Anu French, Scott Shannon, David M. Steinhorn, Minal Vazirani, and Ana Maria Verissimo.The authors would like to apologize for any inconvenience caused to the readers by these changes that do not affect the scientific results. The manuscript will be updated and the original will remain on the article webpage, with a reference to this Correction."} +{"text": "In the original article, we neglected to include the funder \u201cNational Institute of Allergy and Infectious Diseases of the National Institute of Health (NIAID-NIH), R01AI138230\u201d to WD.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "In the Funding section, the grant number from the funder European Research Council is missing. The correct funding information is as follows: This research was funded by CEPREMAP and the European Research Council. Yann Algan received financial support for this work from the European Commission\u2019s Horizon 2020 program under European Research Council Consolidator Grant no. 647870. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Prenatal diagnosis, care and management are involved in mortality and morbidity of every country. A high prevalence is estimated in Africa. We use bibliometrics and mapping tools to explore the area studies and countries involved in scientific research on prenatal diagnosis, care and management in Africa. We used two databases: Web of Science and Pubmed. We extracted sets of data as publication years, organizations, funding agencies, countries from Web of Science core collection database and Medical Subject Headings from Pubmed database. We mapped the data using VOSviewer. We performed keyword analysis. We accessed 463 articles published between 1956 and 2015 in Web of Science Core collection Database and 3372 from Pubmed database. The majority of which were after 2004. The main countries involved in research on prenatal field in Africa were the USA, the United Kingdom, France and South Africa. Two main keywords are relevant: fetal alcohol syndrome and HIV. Prenatal diagnosis, care and management are leaded by South Africa. Some new countries are merging such as Rwanda. The main fields are fetal alcohol syndrome and HIV. It is funded by NIH but also Cape Town University. Maternal and child survival is one of the main goals for the World Health Organization , 2. The We used previously described methods -7. BriefUsing WOS core collection database, we obtained 463 records. More of the records were after 2004 . The maiMost articles were published after 2004 according to the analysis of two databases Pubmed and WOS. This is in accordance with indexation and publication rates of previous studies -7. On thSouth Africa is the main African country involved in prenatal diagnosis, care and management. This is a confirmation of previous general studies on the leading role of South Africa in the continent . In WestKeyword analysis, Web of Science Category analysis and MeSH analysis reveal two main topics: fetal alcohol syndrome and HIV. In fact, public environmental health is one oFetal alcohol syndrome or fetal alcohol spectrum disorders (FASD) are a range of disabilities due to materno-fetal alcohol exposure . Main feThe second main topic is HIV . Old stuIn conclusion, prenatal diagnosis, care and management are leaded by South Africa. Moreover, some new countries are merging such as Rwanda. The main fields are fetal alcohol syndrome and HIV, funded by NIH (USA) but also Cape Town University (South Africa). Coordination of national health policies are required to improve prenatal diagnosis, care and management.The authors declare no competing interest."} +{"text": "In the original article, there was an error in Acknowledgements section. We need to add an acknowledgement of Dr. Robert X. Smith for his contributions towards Figure 1.A correction has been made to the Acknowledgements section.This work was partially supported by the Intramural Research Program of the National Institute on Drug Abuse, the National Institutes of Health (NIH). Data from the Human Connectome Project, WU-Minn Consortium were funded by the 16 NIH Institutes and Centers that support the NIH Blueprint for Neuroscience Research; This work was also supported by NIH grant (UH2-NS100614). The authors are grateful to Drs. Michael Breakspear and Stewart Heitmann for their help with the Brain Dynamic Toolbox. The authors are also grateful to Dr. Robert X. Smith for his contribution of Figure 1.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way.The original article has been updated.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "In addition, the correct number for The National Institute of Biological Resources of Korea is \u201cNIBR201631201\u201d.There is an error in the Funding. The correct name for the funder is \u201cThe National Institute of Biological Resources of Korea\u201d. Website: The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The authors of \u201cVideo-Delivered Family Therapy for Home Visited Young Mothers With Perinatal Depressive Symptoms: Quasi-Experimental Implementation-Effectiveness Hybrid Trial\u201d :e11513) wish to add the following funding statement to their Acknowledgments section:Research reported in this publication was supported by The Dartmouth Clinical and Translational Science Institute, under award number UL1TR001086 from the National Center for Advancing Translational Sciences (NCATS) of the National Institutes of Health (NIH). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.The correction will appear in the online version of the paper on the JMIR website on March 7, 2019, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article also has been resubmitted to those repositories."} +{"text": "Johan Delport is not included in the author byline. Johan Delport should be listed as the seventh author and affiliated with Department of Medical Microbiology, London Health Sciences Center, London, Canada. The contributions of this author are as follows: Methodology and Validation."} +{"text": "Hosung Kang was supported by the Postdoctoral Research Program of Sungkyunkwan University (2015) and Philip M. Graybill was supported by Virginia Tech's BIOTRANS IGEP. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There are errors in the Funding section. The correct funding information is as follows: This work was supported by"} +{"text": "The authors would like to add the funding number to the published article . The corFunding: This research was funded by the Ministry of Education, Youth and Sports of the Czech Republic, agreement number MSMT-5727/2018-2 and by the Ministry of Education and Science of the Russian Federation, agreement No14.587.21.0049 (unique identifier RFMEFI58718X0049).The manuscript will be updated, and the original will remain online on the article\u2019s webpage. The authors would like to apologize for any inconvenience caused to the readers by these changes."} +{"text": "There is an error in the Funding statement. One of the grant numbers is incorrect and should be BE 5601/4-1 instead of BE 5604/1-1.The corrected statement should read: \u201cThis research was supported by German Research Foundation , the Federal Ministry of Education and Research , the National Eye Institute and the Max Planck Society . The funders had no role in study design, data collection and analysis, decision to publish or preparations of the manuscript.\u201d"} +{"text": "AbstractEditorial of special issue of plant diversity in southeastern Asia. BRI) is a long term programme to connect the world with the aim of increasing trade and economic growth and accerelating regional integration. BRI extends into global biodiversity conservation. SE Asia is one of the BRI regions. In order to conserve high biodiversity and promote sustainable development in SE Asia, the Southeast Asian Biodiversity Research Institute (SEABRI) was established by the Chinese Academy of Sciences (CAS), in Nay Pyi Taw in 2015 from the Neogene of South-Western China and Northern Vietnam, a new genus and species of temperate bamboo from Central-Southern Vietnam and a new species of paleotropical bamboo genus Schizostachyum from Vietnam, a new species and two new records of Goniothalamus (Annonaceae) from Laos and studies of Begonia (Begoniaceae) from Laos and Myanmar, a new species of Bulbophyllum (Orchidaceae) from Indonesia, two new species of Alseodaphnopsis (Lauraceae) from South-Western China and Northern Myanmar, a new species of Ainsliaea (Asteraceae) from near the border of Myanmar and China, new species of Colocasia (Araceae), Ophiorrhiza (Rubiaceae), Blumea (Asteraceae) and Zingiber (Zingiberaceae) from Myanmar and taxonomic studies on Amomum (Zingiberaceae) from Myanmar, an annotated checklist of Orchidaceae and notes on Gastrochillus (Orchidaceae) from Myanmar and a new species and two new combinations of Monolophus (Zingiberaceae) from Indo-Burma. All these studies were financially supported by the CAS.The documentation of the rich biodiversity in SE Asia is the very first step towards understanding and conservation of biodiversity . The fir in 2018 . This su"} +{"text": "The following information is missing from the Funding section: This article was funded by the Qatar National Library. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In the original article, we neglected to include the funders Ministry of Trade, Industry and Energy , Ministry of Science and ICT , and Ministry of Health and Welfare , who provided funding under the Technology Development Program for AI-Bio-Robot-Medicine Convergence (20001650).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The following information is missing from the Funding section: This work was supported by Research to Prevent Blindness, New York, New York."} +{"text": "In the version of this article published earlier, the funding information was omitted. The note is hereby provided as: \u2018The authors acknowledge the funding support provided by the Maternal Mental Health Fund, Rural Health Fund, National Research Foundation and the National Health Scholars Programme\u2019. This correction does not alter the study\u2019s findings of significance or overall interpretation of the study results. The publisher apologises for any inconvenience caused."} +{"text": "The following information is missing from the Funding statement: This article was funded by the Qatar National Library. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There are errors in the Funding section. The correct funding information is as follows: This work was supported by the Biotechnology and Biological Sciences Research Council [grant number BB/J015660/1]; and the National Centre for the Replacement, Refinement & Reduction of Animals in Research [grant number NC/N003071/1]"} diff --git a/PMC_clustering_24.jsonl b/PMC_clustering_24.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..74bb9ff102f1df8afc7528e71f475cbf9b66d415 --- /dev/null +++ b/PMC_clustering_24.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:2273eddc94900619e78a295bc2d2a21f4b8e4920bc4d0dfa4d0a96d8aa095132 +size 82495628 diff --git a/PMC_clustering_25.jsonl b/PMC_clustering_25.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..2c221d9604863ad05b66c01ddd134c9a02e40b08 --- /dev/null +++ b/PMC_clustering_25.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid 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--- /dev/null +++ b/PMC_clustering_35.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:ff56b9b9a14b42061dfb6f230e3b94cc0d2f6564d4e62924b724901f79463256 +size 70425292 diff --git a/PMC_clustering_36.jsonl b/PMC_clustering_36.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..638b16ab5e04160db6d2c08fe6e9992945d1bf6f --- /dev/null +++ b/PMC_clustering_36.jsonl @@ -0,0 +1,756 @@ +{"text": "JCI Insight. 2019;4(8):e125172. https://doi.org/10.1172/jci.insight.125172Original citation: JCI Insight. 2020;5(24):e146654. https://doi.org/10.1172/jci.insight.146654Citation for this retraction: JCI Insight of data manipulation in this article. In accordance with the institutional recommendation and at the request of the corresponding author, JCI Insight is retracting this article.Newcastle University recently notified"} +{"text": "R. Soc. Open Sci.7, 200203 (Published Online 11 March 2020) (doi:10.1098/rsos.200203)tER was used in place of t/R (see below). This has now been updated.This correction refers to an error in equation 2.16, Corrected version"} +{"text": "This article has been corrected: During figure processing, identical Western blot images were mistakenly placed in both 7166-7181. https://doi.org/10.18632/oncotarget.3347Original article: Oncotarget. 2015; 6:7166\u20137181."} +{"text": "Apis mellifera L. (Hymenoptera: Apidae) and functional characterization of Am_Eglp 1. PLoS ONE 15(9): e0236724. https://doi.org/10.1371/journal.pone.0236724.The first author\u2019s initials appear incorrectly in the citation. The correct citation is: de Souza DLL, Serr\u00e3o JE, Hansen IA (2020) Aquaporin expression in the alimentary canal of the honey bee"} +{"text": "JCI Insight. 2018;3(21):e97941. https://doi.org/10.1172/jci.insight.97941Original citation: JCI Insight. 2020;5(23):e145847. https://doi.org/10.1172/jci.insight.145847Citation for this corrigendum: The graph shown in The authors regret the error."} +{"text": "R. Soc. open sci.7, 191577. (Published Online 22 January 2020) (doi:10.1098/rsos.191577)This correction refers to an error in the caption for figure 6. The copyright information was missing; this has now been corrected."} +{"text": "Retraction Note: Lab Anim Res (2018) 34: 49\u201357https://doi.org/10.5625/lar.2018.34.2.49This article has beenAll authors agreed to this retraction."} +{"text": "Correction to:Journal of Exposure Science & Environmental Epidemiology10.1038/s41370-018-0110-5published online 10 January 2019In the original Article, References 34\u201336 listed incorrect URLs. The correct URLs are:https://www.health.state.mn.us/communities/environment/biomonitoring/docs/pfc2015communityreport.pdf34. MDH . East Metro PFC3 biomonitoring project: December 2015. Report to the Community. 2015b. http://www.oecd.org/officialdocuments/publicdisplaydocumentpdf/?cote=ENV-JM-MONO(2018)7&doclanguage=en35. OECD (Organization for Economic Co-operation and Development). Toward a new comprehensive global database of per and polyfluoroalkyl substances (PFASs): summary report on updating the OECD 2007 list of per- and polyfluoroalkyl substances (PFASs). 2018. https://19january2017snapshot.epa.gov/expobox/exposure-factors-handbook-2011-edition_.html36. USEPA . National Center for Environmental Assessment. Exposure Factors Handbook. Edition. 2011."} +{"text": "MC1R gene for coat colour variation in Chinese Tan sheep. PLoS ONE 15(8): e0235426. https://doi.org/10.1371/journal.pone.0235426The fourth author's name is spelled incorrectly. The correct name is: Rabiul Islam. The correct citation is: Gebreselassie G, Liang B, Berihulay H, Islam R, Abied A, Jiang L, et al. (2020) Genomic mapping identifies two genetic variants in the"} +{"text": "This article has been corrected: In 7816-7828. https://doi.org/10.18632/oncotarget.6868Original article: Oncotarget. 2016; 7:7816\u20137828."} +{"text": "The Pan African Medical Journal. 2016;23:149. doi:10.11604/pamj.2016.23.149.8950.Cet erratum corrige l\u2019article original: Dans la version originale de l\u2019article, l\u2019orthographe du nom de l\u2019auteur correspondant est incorrecte . Cela a"} +{"text": "The correct name is: Charline M\u00fcntze-R\u00f6hr. The correct citation is: Valentin D, Presas A, M\u00fcntze-R\u00f6hr C, Mele E, Biehl C, Heiss C, et al. (2021) On the quantification of local power densities in a new vibration bioreactor. PLoS ONE 16(1): e0245768. There are errors in the Author Contributions. The correct contributions are: Conceptualization: DV WAB. Data curation: DV. Formal analysis: AP DV. Funding acquisition: AP CH WAB. Investigation: DV AP. Methodology: AP. Project administration: WAB. Resources: CH. Software: DV, CM-R. Supervision: CB CH. Validation: EM CH WAB. Writing\u2013original draft: DV WAB. Writing\u2013review & editing: WAB."} +{"text": "Correction to: Nutr Metabhttps://doi.org/10.1186/s12986-019-0401-4The original version of this article , publish1. Incorrect calculations in Table\u00a02. Correct calculations in Table"} +{"text": "In the original article an authoKhue LM, Jarzabek S. Demonstration paper: mood self\u2013assessment on smartphones. In: Proceedings of the conference on wireless health. ACM, New York, NY, USA. 2015. 10.1145/2811780.2811921.should therefore be corrected as follows:Khue LM, Ouh EL, Jarzabek S. Demonstration paper: mood self\u2013assessment on smartphones. In: Proceedings of the conference on wireless health. ACM: New York; 2015. 10.1145/2811780.2811921."} +{"text": "This article has been corrected: Due to mistakes during the assembly of 94393-94406. https://doi.org/10.18632/oncotarget.21765Original article: Oncotarget. 2017; 8:94393\u201394406."} +{"text": "This article has been corrected: The Grant section information has been updated with the following logos:6494-6508. https://doi.org/10.18632/oncotarget.27303Original article: Oncotarget. 2019; 10:6494\u20136508."} +{"text": "Correction to: J Animal Sci Biotechnol 11, 84 (2020)https://doi.org/10.1186/s40104-020-00482-xIn the original publication of this article , the authttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE145376\u201d should be \u201chttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE145375\u201d.The \u201cThe original publication has been corrected."} +{"text": "R. Soc. Open Sci.7, 200327 (Published online 13 May 2020) (doi:10.1098/rsos.200327)This correction refers to an error in reference [12]. It should be replaced by the below:ReferenceCaretta caretta) and green (Chelonia mydas) sea turtle nests in northern Cyprus. J. Nat. Hist.35, 573\u2013581. (doi:10.1080/00222930151098233)12. McGowan A, Broderick AC, Deeming J, Godley BJ, Hancock EG. 2001 Dipteran infestation of loggerhead (This has now been corrected."} +{"text": "This corrigendum corrects article \u201cExternal auditory canal haemorrhage as the first sign of internal carotid artery pseudoaneurysm: a rare case\u201d and its publication reference i.e. The Pan African Medical Journal. 2020; 37:163. Access corrected manuscript Pan African Medical Journal. 2020; 37: 163. doi: 10.11604/pamj.2020.37.163.21968. PubMed PMID: 33425196. PubMed Central PMCID: PMC7757232. Epub 2021/01/12. eng. The original version of this article had auth"} +{"text": "The fifth author\u2019s name appears incorrectly. The correct name is: P. N. Sylaja. The correct citation is: K. A, Shafeeque CM, Sudhir JB, Banerjee M, Sylaja PN (2020) Ethnic variation and the relevance of homozygous RNF 213 p.R4810.K variant in the phenotype of Indian Moya moya disease. PLoS ONE 15(12): e0243925. https://doi.org/10.1371/journal.pone.0243925The publisher apologizes for the error."} +{"text": "This article has been corrected: Due to errors during typesetting, the image for 66989-67003. https://doi.org/10.18632/oncotarget.11888Original article: Oncotarget. 2016; 7:66989\u201367003."} +{"text": "It also has an archive list to other posts on chemical feedstocks from microbes.https://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-020-01325-0This study focused on using autoclaved municipal waste as a complex feedstock for chemical production, screening different microbes for their ability to transform the mixture.https://www.sciencedirect.com/topics/engineering/feedstocksThis page on feedstocks is a compendium and contains information related to biological and thermo\u2010chemical conversions of biomass.https://www.bio.org/sites/default/files/legacy/bioorg/docs/Synthetic-Biology-and-Everyday-Products-2012.pdfThis site from the Bio manufacturing organization discusses industrial processes for making feedstock chemicals via microbial processes.https://www2.deloitte.com/content/dam/Deloitte/nl/Documents/manufacturing/deloitte-nl-manufacturing-opportunities-for-the-fermentation-based-chemical-industry-2014.pdfThis report focuses on business opportunities for the fermentation\u2010based chemical industries.https://www.genomatica.com/_uploads/pdfs/ISJ_markburke.pdfThis article deals with sustainable manufacture of chemicals with a major focus on Genomatica\u2019s process for converting sucrose to 1,4\u2010propanediol.https://www.riken.jp/en/news_pubs/research_news/rr/20180928_2018fall_FH/index.htmlMaleic acid is an important chemical, used for making surface coatings, resins, lubricants and agricultural products. This article describes a microbial process for producing the chemical.https://www.nature.com/articles/s41929-019-0272-0?draft=marketingWhile there has been much written about using electrochemical processes to drive microbial metabolism, this article looks at the theoretical efficiencies of different processes.https://www.mdpi.com/2227-9717/8/2/199/htmLactic acid is an important chemical feedstock. One output of polylactic acid polymers, and this paper provides a techno\u2010economic assessment.https://www.imedpub.com/articles/exploring-the-microbial-production-of-aromatic-fine-chemicals-to-overcome-the-barriers-of-traditional-methods.pdfde novo synthesis or derived from lignin metabolism.Aromatic compounds may be produced by biological systems, via https://www.pnas.org/content/116/22/10749Escherichia coli and Corynebacterium glutamicum strains.Methyl anthranilate is an important industrial chemical used in foods and cosmetics. This study explored its production in genetically\u2010engineered https://www.aocs.org/stay-informed/inform-magazine/featured-articles/pathways-to-novel-chemicals-february-2014?SSO=TrueThis site discusses the competition between bio\u2010based and new chemical processes for making key industrial specialty chemicals."} +{"text": "Correction to: Cardiovasc Ultrasound (2021) 19:6https://doi.org/10.1186/s12947-020-00235-wFollowing publication of the original article , the autThe incorrect author name is:Marcelo L. C. Viera.The correct author name is:Marcelo L. C. Vieira.The original article has been"} +{"text": "Article title: Caffeine reduces deficits in mechanosensation and locomotion induced by L-DOPA and protects dopaminergic neurons in a transgenic Caenorhabditis elegans model of Parkinson\u2019s diseaseAuthors: Manalo, R. V. M., & Medina, P. M. B.Journal:Pharmaceutical BiologyBibliometrics: Volume 58, Number 1, pages 721\u2013731DOI:http://doi.org/10.1080/13880209.2020.1791192Figure 3E was incorrectly replaced by a re-pasted Figure 3D instead of its original intended figure."} +{"text": "R. Soc. open sci.6, 191304. (Published 11 December 2019). (doi:10.1098/rsos.191304)y-axis in This correction refers to errors in the labelling of the"} +{"text": "Translational PsychiatryCorrection to: 10.1038/tp.2015.136 published online 01 September 2015In the original Article, Figs."} +{"text": "This article has been corrected: The corresponding author, Dianshan Ke, requested to change the email. The correct email is given below:Correspondence to: Dianshan Ke; email: kds8810@163.com . https://doi.org/10.18632/aging.103451Original article: Aging. 2020; 13:13488\u201313501."} +{"text": "Journal of Clinical and Translational Sciences in 2019. The editors, authors and readers are immensely grateful for their thoughtfulness and expertise that have served our journal well.Stephanie AbbuhlTerry AinsworthMyles AkabasTabia AkintobiBianca AlbersEmily AndersonJoseph E. AndrewsJudith AronsonJane AtkinsonMona AuYoungLorena BaccagliniLaura BalisMakenzie L. BarrMark BauerAna BaumannLiza BehrensMiriam BenderL. Michelle BennettJesse David BermanNazleen BharmalArlene BiermanBeatrice A. BoatengKathleen T. BradyPatrick BrandtDonna BrassilTabetha A. BrockmanChad BrummettCheryl BushnellNancy A. Calvin-NaylorJose CancelasMarjory CharlotMichael CharltonPeggy ChenJames J. CiminoRobert A. ClarkBarry CollerLinda CottlerMara CoyleJennifer CrokerOrianna DamasColin A. DeppMegan DoerrAalap DoshiAnn M. DozierMilton Mickey EderVicki L. EllingrodKolaleh EskandanianEstela EstapeAlecia FairStephanie FreelJanice L. GabriloveRussell E. GlasgowMelody GoodmanMary GorfineShelly GrayAlexandra Greenberg-WorisekMatthew GrossmanKristie B. HaddenHeidi HansonBrett HarnettPaul A. HarrisMatthew HartJ.R. HaywoodWilliam R. HoganMike HogarthLianna IshiharaRebecca D. JacksonJason JohnsonRachel JonesYvonne JoostenFelichism W. KaboAllison KarpynHeidi KeelerMatthew KeenerPhilip KernRoohi KharofaDana KingAgnes KiraggaJackie KnapkeRon KoenigH. Robert KolbRhonda G. KostSunil KripalaniJack KuesDanielle LavalleeColleen E. LawrenceScott P. LayneScott LeischowAaron L. LeppinVivian LewisZ.N. LiMike LinkeKanchan LotaMike LyonsJane MahoneyElizabeth MalcolmCamille Anne MartinaGeorge A. MashourPhoenix A. MatthewsColleen A. MayowskiWayne T. McCormackRachel McGarrigleEmma Anne MeagherTara G. MehtaPaul MeissnerJonathan MerrellFrederick J. MeyersPeter MeyersLloyd MichenerChristopher MillerRachel MoonSean MooneyChristopher MorleyCynthia MorrisGia Mudd\u2013MartinDonald E. NeaseJohn OetzelChristian OhmannJanet M. OkamotoJulie OzierDinesh PalAngela L. Palmer\u2013WackerlyCecilia Patino SuttonSusan M PerkinsSharon PlonByron PowellLori Lyn PriceEnola ProctorJill M. PulleyBoriska RabinJulie RainwaterMarie RapeJennifer ReedRobert L. RhyneSuzanne RiveraBrenda RoblesStephen Joseph RosenfeldKristie RossDoris RubioPatrick RyanMaritza Salazar CampoElias SamuelsElaine SchattnerMichael SchembriCari SchmidtEllie SchoenbaumLinda M. SchollHarry SelkerDana ShafferJackilen ShannonAnantha ShekharGreg SimonChristine SorknessKathleen R. StevensWilliam StratbuckerAlisa SurkisAlan TaitEllen TamborVelma ThompsonJonathan TobinJoel TsevatKatherine R. TuttleJason UmansLaurie Van EgerenMelissa Van DykeBoris VolkovRahma WarsameKevin J. WeatherwaxKaren WeaversFern WebbMary Beth WeberTerrence WittHenry Nolan YoungWe are indebted to the expert referees who have volunteered their time to review submissions for the"} +{"text": "This article has been corrected: Due to an error during image processing, frame 6 of tumor #1 (one of the control tumors) in 6269-6282. https://doi.org/10.18632/oncotarget.27268Original article: Oncotarget. 2019; 10:6269\u20136282."} +{"text": "This article has been corrected: In 8974-8987. https://doi.org/10.18632/oncotarget.3291Original article: Oncotarget. 2015; 6:8974\u20138987."} +{"text": "In Malek and Long , owing thttps://doi.org/10.5061/dryad.00000001t"} +{"text": "The first author\u2019s name is spelled incorrectly. The correct name is: Lucia Mart\u00ednez Cuesta.https://doi.org/10.1371/journal.pone.0234939. The publisher apologizes for the error.The first author's surname appears incorrectly in the citation. The correct citation is: Mart\u00ednez Cuesta L, Liron JP, Nieto Farias MV, Dolcini GL, Ceriani MC (2020) Effect of bovine leukemia virus (BLV) infection on bovine mammary epithelial cells RNA-seq transcriptome profile. PLoS ONE 15(6): e0234939."} +{"text": "Cirsium sect. Eriolepis are discussed. The accepted names are: Cirsium echinatum, C. eriophorum subsp. eriophorum, C. eriophorum subsp. spathulatum, C. ferox, C. italicum, C. lacaitae, C. lobelii, C. morisianum, C. scabrum, C. tenoreanum, C. vallis-demonii subsp. vallis-demonii, C. vallis-demonii subsp. calabrum comb. nov., and C. vulgare . Four accepted names are typified by specimens preserved at FI (one lectotype), G (one lectotype and one neotype), P (one lectotype), and by illustrations (two lectotypes). Several other heterotypic synonyms of taxa described from Italy are discussed and six of them are typified. A new combination and status are proposed: C. vallis-demonii subsp. calabrum, based on C. eriophorum var. vallis-demonii f. calabrum.The names of the Italian taxa in Cirsium Mill. (Asteraceae Bercht. & J. Presl.: Cardueae Cass.) is a large genus comprising more than 450 species (as many as 491 according to POWO at atEriole himself . TherefoCi. vallis-demonii (which we split into two subspecies). Taxonomical notes are provided within each entry to justify our choice. In the following account, the accepted names are in alphabetical order; within each of them, the treated homotypic synonyms are listed in chronological order.The taxonomic treatment generally follows , with thCirsium echinatum(1) (Desf.) DC., Fl. Franc., ed. 3, 6: 465. 1815 \u2261 Carduus echinatus Desf., Fl. Atlant. 2: 247. 1799 (basion.) \u2261 Cnicus echinatus Willd., Sp. Pl., ed. 4, 3(3): 1668. 1803.\u2014Lectotype .\u2014& Vald\u00e8s (p. 214)Distribution\u2014Species endemic to western Mediterranean le Cirsium echinatum de M. De Candolle , et le Cirsium italicum du m\u00eame botaniste (Cat. Hort. monsp.), appartiennent \u00e0 notre genre ou sous-genre Eriolepis\u201d. Nevertheless, since Cassini . Cassini , who act Cassini (p. 470)sini (in ) did notCirsium eriophorum(2) (L.) Scop., Fl. Carniol., ed. 2, 2: 130. 1771 \u2261 Carduus eriophorus L. (basion.), Sp. Pl. 2: 893. 1753 \u2261 Cnicus eriophorus (L.) Roth, Tent. Fl. Germ.: 345. 1788 \u2261 Eriolepis lanigera Cass. in Cuvier, Dict. Sci. Nat. 41: 331. 1826, nom. illeg. (Art. 11.4).\u2014Lectotype .\u201cCi. eriophorum subsp. eu-eriophorum var. genuinum Gillot\u201d, Rev. Bot. 12: 360. 1894, nom. inval. (Art. 24.3).\u201cCi. eriophorum subsp. vulgare Petr.\u201d, Biblioth. Bot. 78: 15. 1912, nom. inval. (Art. 26.2).Notes on Ci. eriophorum var. vulgare\u2014The final epithet \u201cvulgare\u201d was associated by Naegelius 3: 367. 1904 (sub var. \u201cferox (DC.)\u201d).\u2014Lectotype .Distribution\u2014Species endemic to eastern Spain, southern France and North-western Italy , s.c., s.n. .\u2014=Notes on Ci. italicum\u2014Candolle 3: 367. 1904.\u2014Lectotype (designated here): [illustration] \u201cPhoenix. Leo. Carduus ferox\u201d in L\u2019Obel , August 1905, G. Rigo .\u2014http://mediaphoto.mnhn.fr/media/14413587569952aN3OdAPGgtS3EPg.=Cirsium ferox L. var. lobelii sensu DC.-\u2003 Ci. eriophorum L. subsp. odontolepis (Boiss. ex DC.) Rouy var. aprutianum Rouy\u201d, Bull. Soc. Bot. France 51: 428. 1904, nom. inval. (Art. 38.2 Ex.1).\u201cNotes on Ci. lobelii\u2014The protologue of this name 3: 367. 1904 \u2261 Ci. eriophorum subsp. morisianum (Rchb.f.) Briq. and Cavill. in Burnat Fl. Alp. Marit. 7: 19. 1931.\u2014Lectotype .Lacaita Notes on Ci. morisianum\u2014The protologue of C. morisianum \u201cCarduus gigas acanthoides tomentosus, pycnopolysphaerocephalus\u201d in Cupani , fine di giugno [[A.] Todaro s.n. .\u2014http://147.163.105.223/zoomify/view_img.asp?ic=10462.=Cirsium giganteum var. macrocephalum Lojac., Fl. Sicul. 2(1): 160. 1903.\u2014Lectotype .\u2014http://147.163.105.223/zoomify/view_img.asp?ic=10357).n et al. (p. 522)=Cirsium gigas var. eriophorum Lojac., Fl. Sicul. 2(1): 160. 1903.\u2014Lectotype .\u2014Image of the lectotype available at http://147.163.105.223/zoomify/view_img.asp?ic=10460. et al. Notes on Ca. scaber\u2014Poiret , G. Moretti s.n. .\u2014https://www.ville-ge.ch/musinfo/bd/cjb/chg/adetail.php?id=335721&base=img&lang=fr).\u2014\u201cCi. insubricum Moretti ex Bertol.\u201d, Fl. Ital. [Bertoloni] 9(1): 25. 1853, nom. inval. (Art. 36.1b).Notes on Ci. spathulatum\u2014The name was published after March 1822 . The protologue also includes a description, a reference to Villars et al. , and a taxonomic note, all in Latin. Pertinent material is unfortunately lacking at BOLO, hosting specimens by Moretti , or in other herbaria linked to Moretti ), relying on a specimen sent from Italy by Duby. The name was somewhat inspired by the Linnaean Ca. eriophorus var. spurius cannot be regarded as a basionym; on the other hand, however, it might be reasonable that the validly published Ca. eriophorus var. spurius is acceptable as basionym of the Candollean name under Art. 41.4. Actually, this article cannot be applied in any case, not even disregarding the taxonomic doubt by Candolle, because Ca. eriophorus var. spurius and Ci. eriophorum var. spurium definitely refer to different taxa, i.e., Ci. \u00d7gerhardtii Schultz and Ci. tenoreanum respectively.Candolle publishespurius spurius . As explained by Lacaita 3: 367. 1904 .\u2014Lectotype . et al. Cirsium eriophorum var. involucratum Coss., p. p. subsp. calabrum (Fiori) Del Guacchio, Bernardo, P.Caputo, Domina & Iamonico comb. et stat. nov. \u2261 Ci. eriophorum var. vallis-demonii fo. calabrum Fiori, Fl. Italia 3: 367. 1904.\u2014Lectotype (designated here): Italy, Calabria, s.d., F.V. Zwierlein s.n. , sub Cirsium valdemonense Loj.)\u2014Notes on Ci. eriophorum var. vallis-demonii fo. calabrum\u2014Fiori Lojacono \\ in tutta la Sila ed a Serra San Bruno\u201d. FI053596 (first sheet) includes a flowering branch with three heads and a label identical to the other, but handwritten by Fiori: \u201cCirsium valdemonense Loj.\\Calabria\u201d. The second sheet bears a further flowering branch, without label. The printed date on the sheet and the adoption of the epithet \u201cvaldemonense\u201d, not employed by Fiori later ; the colour is obviously not well observable in dried material, but it was undoubtedly withish in vivo . We choose FI053596 as the lectotype of the Fiori\u2019s name because it is more complete and bears the handwriting of the author.um\u2014Fiori describe). Fiori intended). Fiori . This lai later . This character is taxonomical relevant, because\u2014excluding obvious and sporadical albino individuals\u2014it is constant within each species Bluff & Fingerh. subsp. nebrodensis (Wagenitz and I.M\u00fcll.) Greuter (endemic to Sicily) vs. subsp. macrocephala Dillenb. and Kadereit ; Anthemis cretica L. subsp. messanensis (Brullo) Giardina & Raimondo (endemic to Sicily) vs. subsp. calabrica (Arcang.) R.Fern. ; Aubrieta columnae Guss. subsp. sicula (Strobl) M.A. Koch, D.A. German and R. Karl (endemic to Sicily) vs. subsp. columnae ; Sesleria nitida Ten. subsp. sicula Brullo and Giusso (endemic to Sicily) vs. subsp. nitida ; Thymus praecox Opiz subsp. parvulus (Lojac.) Bartolucci, Peruzzi and Passal. (endemic to Sicily) vs. subsp. polytrichus (A.Kern. ex Borb\u00e1s) Jalas \u201cCarduus spinosissimus\u201d in Gerbi 41: 331. 1826.\u2014Lectotype ) \u2261 Cnicus lanceolatum subsp. rosani (Ten.) Arcang., Comp. Fl. Ital.: 403. 1882 \u2261 Ci. lanceolatum subsp. rosani (Ten.) Arcang., Comp. Fl. Ital., ed. 2: 723. 1894.\u2014Neotype .\u2014 Lacaita (p. 125)=Cirsium crinitum Boiss. ex DC., Prodr. 7(1): 305. 1838 \u2261 Ci. lanceolatum subsp. crinitum (Boiss. ex DC.) Bonnier & Layens, Tabl. Syn. Pl. Vasc. France: 175. 1894 (cf. p. VIII of the same work) \u2261 Ci. vulgare subsp. crinitum (Boiss. ex DC.) Ar\u00e8nes, Bull. Soc. Fran\u00e7. Echange Pl. Vasc. 1: 21. 1948.\u2014Lectotype .\u2014http://www.ville-ge.ch/musinfo/bd/cjb/chg/adetail.php?id=407321&base=img&lang=fr.& Vald\u00e9s (p. 201)=Cirsium misilmerense Ces, Pass. & Gibelli, Comp. Fl. Ital. 2(21): 483. 1878.\u2014Lectotype (designated here): Italy, Sicilia, Sotto Misilmeri, s.d. s.n. (RO!).\u2014For an image of the lectotype, see =Cirsium cardoleonis Lojac., Fl. Sicul. 2(1): 158. 1903\u2014Lectotype .\u2014http://147.163.105.223/herbarium_vdetails_en2.asp?idmode=simple&id=22320.n et al. (p. 521)=Cirsium dubium Lojac., Fl. Sicul. 2(1): 155. 1903.\u2014Lectotype .\u2014http://147.163.105.223/herbarium_vdetails_en2.asp?idmode=simple&id=22476.=Cirsium lanceolatum var. subbipinnatum Lojac., Fl. Sicul. 2(1): 155. 1903.\u2014Lectotype .\u2014http://147.163.105.223/herbarium_vdetails_en2.asp?idmode=simple&id=22387.=Cirsium lanceolatum var. tenuispinum Lojac., Fl. Sicul. 2(1): 155. 1903 (sub \u201ctenuispinus\u201d).\u2014Lectotype .\u2014http://147.163.105.223/herbarium_vdetails_en2.asp?idmode=simple&id=22379.=Cirsium vulgare var. longespinosum Rouy, Fl. France [Rouy & Foucaud] 9: 21. 1905, nom. illeg. (Art. 52.1).\u2014Lectotype (designated here): Italy, Sicily, Palermo sotto la Grazia, Aug , A. Todaro n. 528 .\u2014http://147.163.105.223/herbarium_vdetails_en2.asp?idmode=simple&id=22375.=Ci. lucanicum Lojac., Nat. sicil. 3: 283. 1884\u2014Type:\u2014Not designated .Ci. lanceolatum var. vulgare Naeg.\u201d, Syn. Fl. Germ. Helv., ed. 2, 3: 990. 1845, nom. inval.\u201cCi. vulgare (Savi) Airy-Shaw\u201d, Repert. Spec. Nov. Regni Veg. 43: 304. 1938, isonym .\u201cCi. vulgare (Savi) Petr.\u201d, Sched. Cirsiotheca Univ. 4: n. 33. 1912, nom. prov. (Art. 36.1). By this provisional name, Petrak indicated the taxon correctly named Ci. italicum [\u201citalicum .Notes on Ca. spinosissimus\u2014In a rare booklet, Gerbi Ten., Gerbi taken directly from the protologue of Ca. spinosissimus by Gerbi ([vulgaris as an avowed substitute (nomen novum) for the later homonym Ca. spinosissimus Gerbi. As a consequence, both Savi\u2019s name (Art. 7.4) and obviously its combination in Cirsium [by Savi by Savi by a diy Gerbi Notes on Ci. rosani\u2014Contextually with the name Ci. lobelii, Tenore [Cirsium, dedicating it to his correspondent Francesco Antonio Rosano (1779\u20131843), who first gathered the plant in Basilicata . The protologue includes a Latin description and the provenance . Also in this case, Lacaita [Ci. vulgare, Ci. rosani can be considered a heterotypic synonym. The plate in Flora napolitana [, Tenore (p. 14) Lacaita (p. 125)politana , publishNotes on Ci. misilmerense\u2014This name was published by Cesati et al. [locus classicus (\u201cSotto Misilmeri (Sicilia)\u201d), and of the unpublished name \u201cCnicus misilmerensis Tineo! ined.\u201d. The exclamation mark infers that the new species was described on the basis of a specimen of Tineo\u2019s seen by Cesati. We found this specimen in the Herbarium Cesatianum at RO. It bears a well-preserved plant and the original label by Tineo \u201cCnicus misilmerensis Tin.! ined.|Sotto Misilmeri|leg. Tineo\u201d. Another interesting specimen by Tineo is at PAL (no. 84936), but possibly it was not examined by the authors of the name. Even if regarded, especially in the past, as a distinct [Ci. misilmerense is nowadays mostly included in the variability of Ci. vulgare subsp. crinitum [i et al. , who repdistinct ,87,88 ordistinct ,89, Ci. no. 8493, but poscrinitum .Notes on Ci. lucanicum\u2014Lojacono Pojero [Ci. lucanicum from Ci. italicum and Ci. lobelii sensu Lojacono (=C. tenoreanum). According to modern views [Ci. lucanicum is a synonym of Ci. vulgare subsp. crinitum, which we include in Ci. vulgare. This statement is possibily based also on the taxonomic doubt expressed by Lojacono, who hypothesises that Ci. lucanicum could be Ci. rosani. Nevertheless, according to the protologue, Ci. lucanicum has not decurrent leaves, and this detail would definitely exclude the synonym. In absence of original material and considering this doubt about the synonymization, we refrain to typify this name at present.o Pojero describeo Pojero wrote \u201cio Pojero . Lojacono Pojero did not rn views ,18, Ci. Notes on Ci. vulgare var. longespinosum\u2014Rouy [Flora Sicula Exsiccata n. 528, a synonym by Lamotte, a diagnosis in French , and some localities (at p. 22). Apparently, Rouy himself attributed the name to Todaro. However, as far as we known, this latter author never employed that epithet, not even in his exsiccata. M. Thi\u00e9baut (LY) (in litt.) informed us that Rouy [Ci. lanceolatum var. horridulum Lam., whose epithet ought to have been adopted at varietal rank, its name is definitely superfluous and then illegitimate under Art. 52.1. Nevertheless, citing Flora Sicula Exsiccata n. 528, Rouy [longespinosum\u201d: LY0718280 and LY0718281. These specimens were collected before the protologue and are undoubtedly original material for the name. However, as a further consequence of citing Flora Sicula Exsiccata n. 528, the specimens of this series number are syntypes, which are preferred material for lectotypification (Art. 9.12), also those not seen by the author himself (Art. 9.4). Therefore, we would propose a pertinent specimen at PAL; other syntypes would be preserved in the herbarium cited by Stafleu and Cowan [Ci. lanceolatum All. var. firmum\u201d (see below for Cn. firmus); it includes a flowering stem of Ci. vulgare .sum\u2014Rouy publishehat Rouy only cithat Rouy cited th28, Rouy actually28, Rouy , p. 22) nd Cowan vulgare ,18. HoweTaxonomy\u2014This is a highly variable species. The infraspecific taxa recognized in modern times by several authors Ar\u00e8nes and C. vulgare subsp. silvaticum (Tausch) Ar\u00e8nes, are only preliminarily accepted by Greuter [s e.g., ,19), i.e, i.eTaxo Greuter , Shin an Greuter and are Greuter ,84,95,96Distribution\u2014Widespread and common from the Mediterranean Basin and Europe to Asia, but naturalized worldwide [orldwide ,98.Habitat\u2014Clearings, riparian vegetation, hedges, fields, very often synanthropic , on rich and nitrified soils [ed soils .Cirsium sect. Eriolepis in Italy allowed us to re-evaluate one neglected taxon and to ascertain most synonymies for the correct interpretation of the names. On the other hand, we showed that, in some cases, previous synonymizations were erroneous or very doubtful. In addition, the examination of the original material of names linked to critical taxa suggests that further research should be carried out before accepting taxonomic conclusions. Finally, some overlooked lectotypifications were brought to light.Nomenclatural studies play a central role in systematics and they should be regarded as essential and preliminary for any taxonomic assessment. On one hand, our contribution on"} +{"text": "This article has been corrected: The 1st affiliation information was presented incorrectly. The proper affiliation 1 is as follows:1Guangxi Key laboratory of Metabolic Diseases Research, Central Laboratory of Guilin No. 181 Hospital, Guilin 541002, Guangxi, P.R. China34506-34519. https://doi.org/10.18632/oncotarget.26138Original article: Oncotarget. 2018; 9:34506\u201334519."} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-020-69140-6, published online 17 July 2020Correction to: This Article contains errors in Reference 20 which is incorrectly given as:et al. Come i bambini pensano alla mente del robot. Il ruolo dell\u2019attaccamento e della Teoria della Mente nell\u2019attribuzione di stati mentali ad un agente robotico. Sistemi intelligenti32, 41\u201356. https://doi.org/10.1422/96279 (2020).Cinzia, D. D. The correct Reference 20 appears below:et al. Come i bambini pensano alla mente del robot. Il ruolo dell\u2019attaccamento e della Teoria della Mente nell\u2019attribuzione di stati mentali ad un agente robotico. Sistemi intelligenti32, 41\u201356. https://doi.org/10.1422/96279 (2020).Di Dio, C."} +{"text": "Radiologia Brasileira is officially communicating the Retraction for extraction of the following article:The Editorial Board of https://dx.doi.org/10.1590/0100-3984.2019.0097.Lee JC, Teles MS. Prevalence of additional primary malignancies detected incidentally on PET/CT. Radiol Bras. 2020;53(1):69. Radiologia Brasileira:Reason: Duplicate publication of an article published in a previous issue of https://dx.doi.org/10.1590/0100-3984.2019.0097.Lee JC, Teles MS. Prevalence of additional primary malignancies detected incidentally on PET/CT. Radiol Bras. 2019;52(5):342. Prof. Edson MarchioriRadiologia BrasileiraEditor-in-Chief,"} +{"text": "This article has been corrected: During image assembly, incorrect data was mistakenly used in 18171-18182. https://doi.org/10.18632/oncotarget.7685Original article: Oncotarget. 2016; 7:18171\u201318182."} +{"text": "This article has been corrected: In 35226-35240. https://doi.org/10.18632/oncotarget.26215Original article: Oncotarget. 2018; 9:35226\u201335240."} +{"text": "Trypanosoma brucei . It . It M. o"} +{"text": "This article has been corrected: Due to errors in image processing, in the top panel of 87638-87646. https://doi.org/10.18632/oncotarget.20936Original article: Oncotarget. 2017; 8:87638\u201387646."} +{"text": "Essays Biochem.https://doi.org/10.1042/EBC20200013Grundy, G.J. and Parsons, J.L. (2020) Base excision repair and its implications to cancer therapy. During the production process errors were introduced into"} +{"text": "Molecular Psychiatry (2017) 22:856\u2013873Retraction to: 10.1038/mp.2016.139 published online 30 August 2016The Editor-in-Chief and publisher have retracted this article after anA. Caccamo, C. Branca and S Oddo agree with this retraction. E. Ferreira has not responded to correspondence related to this retraction."} +{"text": "The COVID-19 pandemic has caused us to operate differently with respect to logistics, but fortunately our reviewers have been willing and able to help our authors by providing fair, thoughtful reviews. For doing this while dealing with all the stresses that the pandemic has rained upon us, this year you have my extra special gratitude and that of our senior editors and editors.Each year at this time, it has been my tradition to provide a brief summary of our progress and to thank those of you who have reviewed manuscripts during the year for your service to mSphere carries with it. I therefore thank all of you who have submitted manuscripts in 2020 for that vote of confidence.Despite the pandemic, our submission numbers have greatly increased . Our team interprets this as meaning that you, our authors, have trust in our process and the quality that the name At the beginning of the pandemic, I offered some tips that I thought would help us support each other and the public whom we serve as scientists . It is mAs the end of the year approaches, I hope that you all have a chance to take some time off to enjoy with your family and friends .mSphere family. Let\u2019s look forward together to 2021.Peter AabyZachary AanderudAmr Mahmoud Abd El-GawadAkio AbeLisa Abernathy-CloseWolf-Rainer AbrahamMark AchtmanFelise G. AdamsMark D. AdamsJoshua N. AdkinsPhilippe V. AfonsoAmeeta K. AgarwalAlain Bernardin AgnememelHector C. AguilarPanblo AguilarNacho AguiloDanielle AhnShimpei AikawaVishukumar AimaniandaGillian M. AirMatthew AkersMustafa AkkoyunluFadhl AlakwaaAshfaqul AlamAna Alastruey-IzquierdoJohn F. AlcornHowbeer Muhamad AliAlexandre AlmeidaUri AlonFrancis AlonzoDavid AlsteensCarmen AmaroJorge AmichKarthik AnantharamanCheryl P. AndamChristopher AndersonMatthew Zack AndersonDan I. AnderssonRaul AndinoDiego O. AndreyElliot J. AndrophyEsther R. AngertJuan AnguitaVijay C. AntharamAlberto AntonelliJoseph AntonyYoshichika ArakawaZachary ArdernAngela Arenas-GamboaSilvia ArgimonJos\u00e9 M. Arg\u00fcelloDavid M. AronoffJyoti AroraGustavo ArrizabalagaMichel ArthurSassan AsgariIna Atr\u00e9eAhmed Sherif AttiaWalter J. AtwoodJennifer M. AuchtungSarit AvraniGordon A. AwandareM. Andrea Azcarate-PerilBrian D. BadgleyVictor M. Baizabal-AguirreLauren O. BakaletzSteven BakerSusan C. BakerMegan Tierney BaldridgeCynthia BaldwinCarl J. BalibarJimmy D. BallardElizabeth BallouLudmila BaltazarDavid A. BaltrusRobert BambaraConnor Gavin George BamfordGert BangeJames D. BangsEdel Figueiredo Barbosa-StancioliAntonio BarraganChristopher F. BaslerChristine Marie BassisPhilippe BastinFlorian F. BauerSal BaxamusaNavid BazghalehBernard W. BeallMarco BecherelliJosh R. BeckStephan BeckerSebastian BehrensDeborah Bell-PedersenGeorge A. BelovSarah Ben MaamarRichard J. BennettTeresa M. BergholzAles BerlecBrent BerwinCharles L. BevinsSinem BeyhanAmin Talebi Bezmin AbadiPurnima BhanotJordan E. BisanzS. BishajitRaphael BisinottoJessica Mary Alice BlairSteven R. BlankeMark R. BleackleyRegina B. BledsoeBradley J. BlitvichPatricia Pringle BloomAntje BlumenthalKasun H. BodawattaJason M. BodilyPatrick BoerlinGregory BokinskyNicholas Andrew BokulichAdrianus C. BoonRay BorrowValeria BortolaiaThomas C. G. BoschAnna BothElsa N. Bou GhanemPhilippe BoulocSebastien BoutinAnna BowmanEric BoydEthna Fidelma BoydTess Elizabeth BrewerShaun R. BrinsmadeJeremy S. BrownJessica C. S. BrownMichael G. BrownJohn H. BrumellJames BrustArielle M. BryanWolfgang BuckelMichelle Monique Conni BucknerCarlos BuenoTucker BurchLindsey R. BurchamCara C. BurnsPhilip Brandon BusbeeSarah N. BussMark J. ButtnerJerry M. BuysseMariana X. ByndlossLauren Byrd-LeotisDavid J. BzikMatthew CabeenKen CadwellLaty A. CahoonRichard A. CalderoneMichael Worth CalfeeCarlos Henrique CamargoAndrew CapaldiAlessandra CarattoliRyan B. CarnegieGiovanna CarpiLaura M. CarrollRonan K. CarrollVern B. CarruthersJosep Casades\u00fasSantiago Castillo-Ram\u00edrezClayton C. CaswellLindsay J. CaverlyRodrigo Cay\u00f4Jiri CernyBaofeng ChaiYunrong ChaiDebopam ChakrabartyC. ChakrabortyDouglas L. ChalkerGary C. ChanJosephine R. ChandlerMichael ChandlerYanjie (Jay) ChaoDominique ChaputSujata S. ChaudhariNeeraj ChauhanDamien ChaussabelFeng ChenTingting ChenXinhua ChenGong ChengJackie K. CheungMichaelle ChojnackiVincent T. K. ChowJoseph A. Christie-OlezaGeorge K. ChristophidesKonstantin ChumakovRichard B. ClarkChristine ClaytonAnna R. CliffeElaine Cloutman-GreenPhillip S. CoburnAdam S. CockrellCarolina CoelhoAlexander M. ColeJ\u00e9r\u00f4me CollemareJames CollinsSean CollomsTeresa M. CoquePierre CornelisBruno CorreiaAlfred Cort\u00e9sTh\u00e9r\u00e8se CoudercVictoria H. CowlingRobert A. CramerJohn K. CraneMax CravenerNicholas CrosslandSean CrossonLiwang CuiPaul J. CullenGordon Fritz CusterWeijun DaiRoss Edgar DalbeyMaura DandriDavid J. DavidoDana A. DavisScott C. DawsonElizabeth N. De GaspariJohn P. DekkerNicholas R. De LayFrank R. DeLeoEmma L. DenhamCharlotte De RudderLalitagauri DeshpandeRik L. de SwartAdam M. DeutschbauerAnthony DeVicoSantosh DhakalVijaykrishna DhanasekaranPatricia I. DiazRobert P. DicksonVincenzo Di PilatoEunsoo DoUlrich DobrindtTobias DoerrYohei DoiMaria Dominguez-BelloJustin J. DonatoTao G. DongAnna Dongari-BagtzoglouLaurent DortetFernanda Fernandes dos SantosZhicheng DouBeno\u00eet DoubletCharles M. DozoisJan Felix DrexlerMarc DroletMilton T. DrottKirk DrueyAnamika DubeyBreck A. DuerkopGary M. 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FredricksMegan FreemanTeresa FrisanRyan FrischFriedrich FrischknechtGeorg FritzChristopher Fr\u00f6hlichSimon FrostYoichi FuruyaIwona GabrielAttila GacserNaren Gajenthra KumarFeng GaoRobert GarceaSarahi L. GarciaRodolfo Garcia-ContrerasGenevieve GarrissBeth A. GarvyBaoxue GeAngie GelliNadezhda GermanRohit GhaiMahmoud A. GhannoumNicole GilbertSabine GilchSteven R. GillPeter GilliganRobert D. GilmoreJanet R. GilsdorfNicolas GischLaura GlenndinningErin S. GloagMatthew Robert GoddardRichard GoeringGustavo H. GoldmanStephen GoldsteinJonathan Louis GolobMaria G\u00f3mez Brand\u00f3nNardhy Gomez-LopezBen-Qiang GongChengliang GongJesus Gonzalo-AsensioUri GophnaIsabel GordoClaire L. GorrieRia GoswamiYonatan H. GradIan GraingeLisa GralinskiDaniel A. GreenStefan J. GreenChris GreeningTrudy H. GrossmanVesna GrujcicMarc-Jan GubbelsClaudia GuldimannOzan GundogduArthur G\u00fcnzlXiaofeng GuoTed HackstadtLubomir HadjiskiMarisa HaenniMartin W. HahnConstantine HaidarisAnders P. HakanssonRebecca Jane HallRuth M. HallLuanne Hall-StoodleyRichard HamelinNeal D. HammerKirsten K. HansonNancy D. HansonClare HardingJ. Marie HardwickChristopher HarmerEva HarrisJason B. HarrisErin HarveyCynthia Y. HeMichael HeckerNagendra R. HegdeJacob Heilmann-ClausenBernd HeimrichJon HeinrichsJohannes HelderRichard F. HelmYosra A. HelmyEls HenckaertsVincent Herv\u00e9Mark C. HerzbergMatthias HessSteven HigginsPenelope HiggsJulian F. HillyerPhilip HinchliffeB. Joseph HinnebuschFlorian HladikRichard L. HodinkaSabine Hofmann-ThielJeffrey HollomonCorey C. HoltKathryn Elizabeth HoltYoshikazu Honda-OkuboPeiying HongDavid C. HooperTomoyuki HoriBranka HorvatGuy HouillonKai HuQinxue HuSarah HuAndrew D. HuangLaibin HuangLili HuangWeishan HuangDiego HuetWilhelmina HustonTu Anh N. HuynhJennifer HydeFlorenzo IannoneKensuke IgarashiMolly A. IngersollHanne IngmerRonald M. IorioDaniel IrimiaSuzanne Lynn IshaqWataru IwasakiAngelo IzzoMary Ann Jabra-RizkWilliam R. JacobsKaty JeannotVicki JeffersMatthew L. JeniorVeronica JimenezDong-Yan JinRongsheng JinChristian JobinSophia JohlerJeremiah G. JohnsonPatricia J. JohnsonTimothy J. JohnsonBradley D. JonesJennifer JonesHoward S. JudelsonSandra JunglenCatherine JusteVladimir R. KaberdinDavid KadoshBj\u00f6rn F. C. KafsackPeter C. KahnMarkus KainulainenMaria KalamvokiLindsay KalanMarkus KalkumSaid KamelRobert W. KaminskiBeate KampmannShyam KandelManabu KannoStefan KappeNabil KarahPetros C. KarakousisHazma Umut KarakurtLisa KarstensYuichiro KashiyamaTsutomu KatayamaSophia KathariouSouichiro KatoCarol A. KauffmanGhazi KayaliHangjun KeBrendan KellyEllen KenchingtonLinda J. KenneySuliman KhanSahil KhannaMegan R. KiedrowskiNicolas KiefferHye Kwon KimJae-Ouk KimLee KimbellJanine KimpelGary M. KingSamantha Jane KingJoseph A. KirkLea-Ann S. KirkhamLaura A. KirkmanTodd KittenHiroshi KiyonoMichiel KleerebezemBruce S. KleinAndreas KlosLaura J. KnollDennis C. KoWolfgang KoesterMichael H. KogutAlain KohlAbimbola KolawoleNicholas KomarHeidi KongJames B. KonopkaEugene V. KooninIngo K\u00f6perMilena KordalewskaOmry KorenNicole Marie KoropatkinNatalia KorotkovaAnita A. KoshyTakahiko KoyamaLukasz KozubowskiFlorian KrammerLaurent KremerEric S. KrukonisMichael D. KruppaJason KubinakKengo KubotaAndreas KuhnJens H. KuhnEric KuhnertAnuj KumarBeate Mareike K\u00fcmmererHarry D. KurtzHiroyuki KusadaJoseph L. KutiSebla Bulent KutluayJennie H. KwonMonique LafonEric R. LafontaineRenee M. LairdLaura LambTilman LamparterStephanie LangelMary L. Lap\u00e9-NixonMarilynn A. LarsonChristian LauberAdam S. LauringCatherine LavazecAllen LeeSamuel A. LeeSoo Chan LeeVincent T. LeeYong-Hwan LeeMcKenzie K. LehmanBenfang LeiJ\u00f8rgen J. LeisnerChristopher W. LennonDion LeppJhansi L. LeslieJohan LeveauSteve LeverCeline M. LevesqueZachary A. LewisBin LiLin-Xi LiMin LiPeng LiZiyin LiXiao-Ping LiaoShen Jean LimDominique H. LimoliAndrew H. LimperJonathan Y. LinMichael LinNilton LincopanJodi A. LindsayZhuoren LingJacqueline Callihan LinnesMartin LinsterPeter N. LipkeTingli LiuZhixia LiuMar\u00eda A. LlamasKaren G. LloydMichael K. LoNaeemah LoganSarah LondriganAllison LopatkinJose L. Lopez-RibotAlessio LorussoSebastian LouridoShirley LuckhartRuth Ann LunaXin M. LuoKalpana LuthraJoseph LutkenhausHinh LyMichael LyuBing MaLi-Jun MaMilton MacielErich R. MackowCalman A. MacLennanEric MaiMichael H. MalamyVera ManageiroMelissa ManusJennifer A. ManuzakRichard T. MarconiRachel Leah MarineKevin MaringerSara MartiEmily Toth MartinJavier MartinLuis R. MartinezJose-Luis Martinez-GonzalezThorsten MascherS\u00e9bastien MatamorosCyrille MathieuJyl S. MatsonJelle MatthijnssensJoseph McBrideShonna M. McBrideBruce A. McClaneAndrea McCollumJohn K. McCormickClaire E. McCoyJohn T. McCroneLarry S. McDanielDiane McDougaldAnita K. McElroyNoel G. McElvaneyQuinn S. McFrederickPatrick McGannMartin J. McGavinJames B. McKinlayRobert J. McLeanDavid N. McMurrayRyan Philip McNamaraGareth M. McVickerAnita F. MeierOlivier MeilhacAsuncion MejiasRoberto G. MelanoJay MelliesEsther MenendezPaola E. MeraTod J. MerkelCraig MeyersMatthew MikoleitLaura MilazzoAndrew MillardYves MillemannAaron W. MillerAlita A. MillerChad MireYoshiyuki MishimaBibhuti MishraNagendra N. MishraSatoshi MitaraiEliane Namie MiyajiKazufumi MochizukiLuke A. MoeAndrew MoellerJeffrey MoffitMaria F. MojicaIstvan MolnarJonathan MonkDavid P. MooreRobert J. MooreNathaniel John MoormanLu\u00eds Fernando de Sousa MoraesTiago Facury MoreiraHiroshi MoriJoachim Morschh\u00e4userMary MotylQu\u00e9zia MouraPaula J. MouserAhmed M. MoustafaW. Scott Moye-RowleyMonica MugnierVasant MuralidharanIsabel Muro-PastorEain A. MurphyTimothy F. MurphyMustapha M. MustaphaTin Tin MyaingThierry NaasAnusha NaganathanMoon H. NahmRyosuke NakaiCindy H. NakatsuKoji NakayamaFranz NarberhausFarooq NasarLu\u00eds Cl\u00e1udio Nascimento da SilvaSonia Navas-MartinErin M. NawrockiDavid M. NeedhamCassandra E. NelsonUjjwal NeogiJeniel E. NettBenjamin W. NeumanFelipe Piedade Gon\u00e7alves NevesStephanie L. NevilleDuane W. NewtonIrene L. G. NewtonCheryl A. NickersonMarisa Fabiana Nicol\u00e1sAngela M. NiliusAngela Helen NobbsPeter A. NobleCarolina Silva NodariMarc Noguera-JulianRomolo NonnoPatrice NordmannSteven J. NorrisJeanette M. NortonDominik N\u00f6rz\u00c2ngela NovaisMairi C. NoverrTomoyoshi NozakiTakuro NunouraBrian B. OakleyJoshua ObarShelby L. O\u2019ConnorDavid O\u2019DwyerPeter OelschlaegerJames P. O\u2019GaraNaoya OharaShigefumi OkamotoYusuke OkazakiFaten OkdaAndrew J. OliveMichael OlsonTeresa O\u2019MearaTroy O\u2019NeilEng Eong OoiAndres Opazo-CapurroWilliam D. OrsiEric OswaldGeorge O\u2019TooleC. Mark OttElizabeth A. OttesenMarc OuelletteMark S. PagetTanapat PalagaRaghavan U. PalaniappanMitchell PallettSatheshkumar PanayampalliJohn C. PanepintoCostas C. PapagiannitsisDaniel Paredes-SabjaJoanna L. ParishDan Mcfarland ParkHee-Soo ParkJason ParkDane ParkerDonovan H. ParksColin R. ParrishChristopher M. ParrySally R. PartridgeErica PasiniDavid PatersonTimothy C. PaulitzMatthew PaulyJovan PavlovicHelene M. M. PaxtonAndrew PekoszXinxia PengJose Christian PerezBenjamin PerinAndreas PeschelBrian PetersFelicitas PfeiferMrudula PhadkeBirgit PiechullaAndrzej PiekarowiczShmuel PietrokovskiTatiana C. A. PintoJohann PitoutGregory V. PlanoRandall PlattDaniel PletzerCarolina H. PohlLaurent PoirelChristopher Robert PolageShawn W. PolsonFrederic PolyMar\u00eda Teresa Ponce-NoyolaDavid L. PophamErin P. PriceSean T. PriggeHarry E. PrincePeter M. PryciakMichael J. PucciAlexandra E. PurdyNicole E. PutnamDohun PyeonLizeng QinJianming QiuPilar QuintanaRobert G. QuiveyBrent RaceGovindarajan RajagopalanMichael RakSrinivasan RamakrishnanKumaran S. RamamurthiChristophe Rami\u00e8reMaria Soledad RamirezCayo RamosChad A. RappleyeDavid RaskoAdam J. RatnerR. S. RedmanPeter ReevesBarbara RehermannAaron ReinkeLina ReslanPeter ReutherMatthew R. ReynoldsTodd B. ReynoldsJonathan RichardsBert K. RimaMarilyn C. RobertsDerrick R. RobinsonMarcio L. RodriguesEstefania RodriguezGeraint B. RogersRobin Rebecca RohwerMohsen RokniSandra Romero-SteinerR. Martin RoopAlexandre Soares RosadoJason W. RoschJean-Pierre RoutyEmeline RouxCraig R. RoyNicole RoyPolly RoyAlex RubinsteynChristian J. RudolphCristian RuizOlena RzhepishevskaWilmara Salgado-Pab\u00f3nAmali SamarasingheLinus SandegrenFelipe H. Santiago-TiradoEzequiel SantillanClarissa Santos RochaToyotaka SatoCharles A. ScangaJoy ScariaJeffrey W. SchertzerPatrick D. SchlossEric SchmidtJennifer Elise SchmidtThomas Mitchell SchmidtVolker SchmidtMirco SchmolkeTony SchountzMichael SchuitChristian SchwerkJarrod J. ScottAmin SedokaniAnna Maria SeekatzJulie A. SegreKate L. SeibH. Steven SeifertRangaraj SelvaranganHidenobu SenpukuKeun Seok SeoAswin Sai Narain SeshasayeeWilliam M. ShaferDilip ShahPriya S. ShahShiraz A. ShahKathy Ho Yen ShairDipali SharmaJessica R. SheldonAimee ShenBang ShenMang ShiShinsuke ShigetoShin-Ru ShihTeppei ShimiasakiDaisuke ShiomiNahum Y. ShpigelJoshua D. ShroutShahid SiddiqueAnita SilRoberto SilvaLynn L. SilverMonique SimierAnthony P. SinaiSteven M. SingerUpinder SinghAlbert SiryapornJerod A. SkybergRenata Dezengrini SlhessarenkoMonika S\u0142omi\u0144ska-Wojew\u00f3dzkaCarolyn M. SlupskyMark S. SmeltzerDarian SmercinaDarci SmithJames Leif SmithJoseph D. SmithWiep Klaas SmitsTeemu SmuraEvan S. SnitkinJonathan W. SnowMilena SokolowskaEvgeni SokurenkoDavid R. SollHarini SooryanarainJoseph A. SorgDaniel StadlbauerRenske D. M. SteenbergenLisa Y. SteinEike SteinmannDavid Cole StevensDennis StevensBrian StevensonDavid B. StewartScott StibitzAshley L. St. JohnGregory G. StoneDaniel StraumeFrank StubenrauchKarina StuckenJorg StulkeCarlos S. SubausteDavid SuePaul M. SullamJie SunShan SunMaarit SuomalainenIvan SurovtsevMichael SurretteJoyce SutcliffeMehul S. SutharTroy C. SuttonElena SuvorovaStaffan G. Sv\u00e4rdMaxim S. SvetlovJoel SwansonW. Edward SwordsFrancois-Etienne SylvainRita TamayoMing TanBenjamin TangYuyang TangArnaud TatonMiguel Cacho TeixeiraAkihiko TeradaBenno Herman Ter KuileKen TeterRajani ThanisseryKevin R. TheisCasey M. TheriotGavin H. ThomasElizabeth ThrallThomas ThurnheerAndrea TicinesiRafal TokarzAndrew TomarasSteven Y. C. TongOlivera Topalovi\u0107Tadashi ToyamaStephen TristramNicolas TromasFran\u00e7ois TrotteinTakafumi TsuboiClaire Elizabeth TurnerKenneth L. TylerGregory TyrrellJuan Esteban UgaldeDavid UlaetoImran UllahGottfried UndenPriya UppuluriSyun-Ichi UrayamaOlivier VallonMiguel A. ValvanoChris Van BenedenKurt Jason VandegriftDan VanderpoolAdrianus W. M. van der VeldenPatrick Van DijckMark W. J. van PasselDebby van RielVicky L. van SantenBrian D. VanScoyDaria Van TyneArvind VarsaniJoel Vega-Rodr\u00edguezLonneke VerveldeAlejandro J. VilaVladimir VinnikJoerg VogelChantal B. VogelsRobin Voigt-ZuwalaPetr VolfVeronika von MesslingJay VornhagenDaniel E. VothJo-Marie VreulinkJatin M. VyasJoseph Thomas WadeRezwanul WahidMatthew K. WaldorSeth T. WalkLouise A. WalkerMark J. WalkerDaniel WallNicholas A. WallaceJudd WalsonJens WalterHui WangJincheng WangJoyce WangJue D. WangXiaoxue WangYan WangYang WangJonathan Mark WarawaMatthew J. WargoChristopher M. WatersNadeeka Kumari WawegamaKeith E. WeaverRichard J. WebbyXin WeiBrian C. WeinrickLouis M. WeissEric WenzlerDawn WetzelNicole WheelerStephen C. WhissonGregory WhitakerJason K. WhitmireGottfried WilharmJulia WillettEmma H. WilsonRichard A. WilsonSteven S. WitkinChristiane E. WobusAlan J. WolfeMichael H. WoodworthKaren WozniakRachel A. F. WozniakGerard D. WrightNicholas C. WuWilliam H. WunnerSusan WyllieKelly Leanne WyresHan XiaHang XieJin-Rong XuXiyuan XuYuquan XuChaoyang XueTimothy L. YahrS. Steve YanHee-Jeong YangYang YangYe YangZhaomin YangCatherine YoshidaYang YuYunsong YuMin YueJoseph P. ZackularRahat ZaheerKamarul ZarkasiRaffaele ZarrilliMichael ZasloffAnna C. ZemkeGabriel E. ZentnerBing ZhaiYuanchao ZhanCheng-Cai ZhangChiyu ZhangRui ZhangJiangchao ZhaoBeiwen ZhengHao ZhengGuangming ZhongXiaohui ZhouRan ZichelLisa ZieglerSeth ZostAbdelrahman ZueterAgain, my sincere thanks to all of you for being part of the"} +{"text": "The Aberdare Ranges Forest, located in the Central highlands of Kenya, is an isolated volcanic mountain in the East African Rift Valley with unique flora. Despite its refugial importance to rare and endemic plant species, the diversity of plants in the Aberdare Ranges Forest remains poorly understood. The checklist presented here is a collation of data obtained from multiple floristic surveys and from herbarium specimen collections from the forest. A total of 1260 vascular plants taxa representing 136 families, 613 genera, 67 subspecies and 63 varieties are documented. The ferns comprised 84 species, lycophytes seven, gymnosperms six and angiosperms were 1163 taxa. This represents 17.9% of the Kenyan taxa, 1.7% of the African taxa and 0.3% of all the vascular plants known in the world. A total of 18 taxa were endemic and 14 taxa were found to be threatened globally. The life form, voucher specimen(s), habitat and distribution range of each taxon and a brief analysis of taxa diversity is presented in this checklist. This is the first comprehensive inventory of vascular plants in the entire Aberdare Ranges, providing a solid basis for more sustainable management and improved conservation of this montane forest. The checklist is also an important contribution to the world checklist of plants required by the Global Strategy for Plant Conservation. EABH) is amongst the eight known hotspots in Africa with globally significant diversity and endemism , through the Global Strategy for Plant Conservation (GSPC), has over the past two decades advocated for intense exploration and documentation of plants species with the aim of achieving a complete world checklist of flora in the near future Forest, previous floristic studies have either focused on a single or a few selected taxa document the vascular plants of the entire AR Forest, (ii) document the endemic and threatened vascular plants in the AR Forest and (iii) document the life forms and habitats of all the vascular plants in the AR Forest.The goal of this study was to provide a broad checklist of vascular plants in the entire AR, together with Mount Kenya, constitute the Central Highlands of Kenya. It extends approximately 120 km southwards from the equator through Nyeri, Nyandarua, Muranga and Kiambu Counties to the Kikuyu escarpment, between 36\u00b030'E, 0\u00b005'S and 36\u00b055'E, 0\u00b045'S AR, togetlevation . The AR f 565 km .AR is diverse and comprises numerous undulating hills formed through volcanism and faulting of the earth\u2019s surface from the early Tertiary to the Pleistocene periods , which manages the Aberdare National Park with an area of ca. 76,700 ha and the Kenya Forest Service (KFS) which governs the Aberdare Forest Reserve measuring ca. 139,500 ha. The National Park encompasses the summit of the AR above 3000 m a.s.l. and a narrow salient extending to the east to 1900 m a.s.l. and Sino-Africa Joint Research Center (SAJOREC) carried out field investigations from 2016 to 2019. Floristic surveys were done during both wet and dry seasons to capture a wide range of phenological cycles of taxa, especially flowering and fruiting. General walk-over surveys were used in specimen collection and habitat characterisation (East African herbarium (EA). Standard botanical references were used in identification of specimens, i.e. Flora of Tropical East Africa in China. Herbarium acronyms follow Thiers (2020 onward: http://sweetgum.nybg.org/science/ih/).A botanical team from the t Africa , BlundelAR and deposited in the EA, were compiled with our collections to develop a checklist of vascular plants. Habitat(s) and relative distribution range for each taxon were determined using our collections, herbarium specimens in the EA and published bibliographies. The broadest altitude ranges, that is between the minimum and maximum altitude a taxon is known to occur, were searched and recorded. Thence, the distribution ranges of plants\u2019 taxa were not restricted to the AR as some taxa had wide distribution ranges extending to the sea level. Life forms of taxa collected were categorised as herbs , shrubs (plants between 50 cm to 5 m high with woody stems branching at or near the ground), climbers (plants with twining herbaceous or woody stems) and trees (GBIF) (https://www.gbif.org). Moreover, the conservation status of all the vascular plants recorded were assessed in the IUCN Red List of Threatened Species (https://www.iucnredlist.org) and categorised as Critically Endangered (CR), Endangered (EN), Vulnerable (VU) and Near Threatened (NT). The current taxonomic circumscription of each taxon recorded was checked in the Tropicos database (http://www.tropicos.org/), African Plant Database (http://www.ville-ge.ch/musinfo/bd/cjb/africa/recherche.php?langue=an) and the Catalogue of Life, 2019 Annual Checklist . Finally, the recorded plants\u2019 taxa were grouped into their respective classes and families and presented alphabetically.Vascular plants specimens, previously collected for varied purposes from the n trunk) . Standarn trunk) , BlundelAR. Ferns and fern-allies were 91 in total, with 1169 taxa of seed plants. The most diverse class was Magnoliopsida , followed by Liliopsida (19.4%), then Polypodiopsida (6.6%), Lycopodiopsida (0.6%) and the least diverse was Pinopsida (0.5%) of the total taxa recorded , Poaceae (8.2%), Fabaceae (6.6%) and Lamiaceae (3.9%) of the total vascular plants recorded (Table Cyperus (20), Helichrysum (19), Senecio (17), Asplenium (17), Crotalaria (15) and Solanum (15), while other genera had less than 14 taxa and Apiaceae (three) and Cyperaceae (three taxa). The majority of the threatened taxa were herbs (nine), then trees (three) and the remaining were shrubs (two taxa).A total of 13 taxa were found to be threatened or near-threatened globally in the Appendix . The taxAR Forest is a significant regional centre of plant diversity. With a total of 1,260 taxa recorded, it represents 17.9% of the total 7,004 vascular plants in Kenya, 10.2% of the 12,317 vascular plants in East Africa, 1.7% of the estimated 74,000 taxa in Africa and 0.3% of the estimated world flora of 422,127 taxa Kalkm and others, are traditionally utilised by the local community as remedies for various illnesses Kalkm and Crotonalienus Pax are faced with logging and charcoal burning threats, thus, they should also be prioritised in conservation planning are endemic while those with (*) are exotic and/or naturalised in the AR Forest. For each taxon recorded, full authority is given, life form, brief notes on habitat and distribution range, voucher specimen number and the herbarium where it was deposited. The broadest altitude range of each taxon is indicated in metres (m) which mostly extends beyond the elevation of the AR Forest. EA refers to the East African herbarium in Nairobi, Kenya, while HIB refers to Wuhan Botanical Garden herbarium in Wuhan, China. The collectors are abbreviated as follows: SK means Solomon Kipkoech, SAJIT refers to Sino-Africa Joint Investigation Team, FOKP means Flora of Kenya Project, KEFRI refers to Kenya Forestry Research Institute and EANHS stands for East Africa Natural History Society.An annotated checklist of the vascular plants of the I system , Pinopsi6 system . Taxa prLycopodiaceaeF1. Austrolycopodiumaberdaricum (Chiov.) Holub \u2013 Life form: Herb. Habitat: Upper parts of montane forest, 3000 m. Voucher: Balbo 475 (EA).Lycopodiumclavatum L. \u2013 Life form: Herb. Habitat: Moist montane forest, 1500\u20133050 m. Vouchers: Mutangah 2 (EA), SK 0148 .Phlegmariurusdacrydioides (Baker) A.R.Field & Bostock \u2013 Life form: Herb. Habitat: Woodland and riverine forest, 1550\u20132700 m. Voucher: Faden 69/1113 (EA).Phlegmariurussaururus (Lam.) B.\u00d8llg. \u2013 Life form: Herb. Habitat: Near streams and damp sites in moorland, 2200\u20134400 m. Voucher: Hedberg 1627 (EA).Phlegmariurusverticillatus (L.f.) A.R.Field & Testo \u2013 Life form: Herb. Habitat: Moist woodland and wet forest, 950\u20132300 m. Vouchers: Someren s.n., Balbo 819 (EA).SelaginellaceaeF2. Selaginellagoudotianavar.abyssinica (Spring) Bizzarri \u2013 Life form: Herb. Habitat: Near waterfalls and riverbanks in evergreen forest, 750\u20132450 m. Vouchers: Ng\u2019weno 15129, Kuchar 12753 (EA).Selaginellakraussiana (Kunze) A.Braun \u2013 Life form: Herb. Habitat: Moist forest, 1100\u20133350 m. Voucher: Lind and Agnew 5013 (EA).AspleniaceaeF3. Aspleniumabyssinicum F\u00e9e \u2013 Life form: Herb. Habitat: often epiphytic in moist forest and damp sites in moorland, 1350\u20133150 m. Voucher: Kamau 364 (EA).Aspleniumactiniopteroides Peter \u2013 Life form: Herb. Habitat: Rocky sites in upland forest, 2500\u20134250 m. Voucher: Someren 1054 (EA).Aspleniumadamsii Alston \u2013 Life form: Herb. Habitat: Wet rocky sites in moorland and heath zone, 2400\u20133400 m. Voucher: Polhill 12026 (EA).Aspleniumaethiopicum (Burm.f.) Bech. \u2013 Life form: Herb. Habitat: Epiphytic in moist forest and wooded grassland, 1150\u20133700 m. Voucher: Mutanga 13 (EA).Aspleniumboltonii Hook. ex Schelpe \u2013 Life form: Herb. Habitat: Epiphytic in moist forest, 1200\u20132750 m. Voucher: Robertson et al. 3895 (EA).Aspleniumbugoiense Hieron. \u2013 Life form: Herb. Habitat: Upland moist forest and along streams, 1650\u20132700 m. Voucher: Faden et al. 74/1346 (EA).Aspleniumelliottii C.H.Wright \u2013 Life form: Herb. Habitat: Moist montane forest, 1050\u20133000 m. Voucher: Kuchar 12407 (EA).Aspleniumerectum Bory ex Willd. \u2013 Life form: Herb. Habitat: Epiphytic in upland forest floors, 1300\u20132750 m. Voucher: Faden 69/004 (EA).Aspleniumfriesiorum C.Chr. \u2013 Life form: Herb. Habitat: Epiphytic in moist forest, swamps and along streams, 1100\u20133000 m. Voucher: Mutanga 10 (EA).Aspleniumlinckii Kuhn \u2013 Life form: Herb. Habitat: Shady places in moist forest, 1600\u20132700 m. Voucher: Faden 74/1317 (EA).Aspleniumloxoscaphoides Baker \u2013 Life form: Herb. Habitat: Epiphytic in moist montane forest and Hagenia woodlands, 1850\u20133650 m. Voucher: Kuchar 5218 (EA).Aspleniummonanthes L. \u2013 Life form: Herb. Habitat: Moist forest and bamboo thickets, 1950\u20133400 m. Voucher: Kamau 366 (EA).Aspleniumpraegracile Rosenst. \u2013 Life form: Herb. Habitat: Moist montane forest and bamboo zone, 2400\u20133100 m. Voucher: Faden et al. 71/880 (EA).Aspleniumrutifolium (P.J.Bergius) Kunze \u2013 Life form: Herb. Habitat: Epiphytic in moist forest and riverine forest, 750\u20132300 m. Voucher: Faden 69/2075 (EA).Aspleniumsandersonii Hook. \u2013 Life form: Herb. Habitat: Epiphytic in moist montane forest and forest margins, 680\u20133100 m. Voucher: Faden 69/012 (EA).Aspleniumtheciferum (Kunth) Mett. \u2013 Life form: Herb. Habitat: Epiphytic in moist forest and bush thickets, 850\u20132900 m. Voucher: Kibui 50 (EA).Aspleniumuhligii Hieron. \u2013 Life form: Herb. Habitat: Epiphytic in upland woodland, 2400\u20134200 m. Voucher: Kuchar 13077 (EA).AthyriaceaeF4. Athyriumnewtonii Baker \u2013 Life form: Herb. Habitat: Upland rocky forest margins, 1150\u20133500 m. Voucher: Tweedie 1882 (EA).Athyriumscandicinum (Willd.) C.Presl \u2013 Life form: Herb. Habitat: Moist forest and bamboo zone, 1150\u20133500 m. Voucher: Faden 70/63 (EA).Depariaboryana (Willd.) M.Kato \u2013 Life form: Herb. Habitat: Upland moist forest, 1460\u20132550 m. Voucher: FOKP 1983 .BlechnaceaeF5. Blechnumaustrale L. \u2013 Life form: Herb. Habitat: Mixed bamboo forest and wet grassland, 1500\u20132500 m. Vouchers: Cameron 18, Gilbert 6315 (EA).Blechnumtabulare (Thunb.) Kuhn \u2013 Life form: Herb. Habitat: Mixed bamboo forest and wet grassland, 1600\u20132600 m. Voucher: Gilbert 6337 (EA).CyatheaceaeF6. Cyatheamanniana Hook. \u2013 Life form: Tree. Habitat: Moist forest and along streams, 1500\u20132500 m. Voucher: Napper 721 (EA).CystopteridaceaeF7. Cystopterisfragilis (L.) Bernh. \u2013 Life form: Herb. Habitat: Damp rocky sites in evergreen forest, 1700\u20133600 m. Voucher: Fries & Fries 762 (EA).DennstaedtiaceaeF8. Blotiellaglabra (Bory) R.M.Tryon \u2013 Life form: Herb. Habitat: Moist forest, 1350\u20133000 m. Voucher: Faden 71/202 (EA).Hypolepisgoetzei Reimers Life form: \u2013 Herb. Habitat: Moist forest, 2100\u20133050 m. Voucher: Faden 71/886 (EA).Hypolepissparsisora (Schrad.) Kuhn \u2013 Life form: Herb. Habitat: Moist forest, 900\u20132800 m. Voucher: Verdcourt 3989 (EA).Pteridiumaquilinum (L.) Kuhn \u2013 Life form: Herb. Habitat: Wooded grassland and forest margins, 2640\u20132850 m. Vouchers: Otieno 12609, Kuchar 7827 (EA).DryopteridaceaeF9. Arachniodeswebbianavar.foliosa (C.Chr.) Gibby, Rasbach, Reichst., Wid\u00e9n & Viane \u2013 Life form: Herb. Habitat: Moist forest and streams banks, 1380\u20132600 m. Vouchers: Gardner 967, Kamau 471 (EA).Dryopterisantarctica (Baker) C.Chr. \u2013 Life form: Herb. Habitat: Montane grassland and moorland, 2500\u20133320 m. Voucher: Faden 69/803 (EA).Dryopterisfadenii Pic.Serm. \u2013 Life form: Herb. Habitat: Moist forest and riverine forest, 1700\u20132500 m. Voucher: Faden et al. 69/900 (EA).Dryopterislewalleana Pic.Serm. \u2013 Life form: Herb. Habitat: Moist forest and riverine forest, 1400\u20132300 m. Voucher: Faden 70/55 (EA).Dryopterismanniana (Hook.) C.Chr. \u2013 Life form: Herb. Habitat: Moist forest, 1450\u20132250 m. Voucher: Faden et al. 69/289 (EA).Elaphoglossumangulatum (Blume) T.Moore \u2013 Life form: Herb. Habitat: Epiphytic in moist montane forest, 2470\u20132750 m. Voucher: Faden 71/201 (EA).Elaphoglossumaubertii (Desv.) T.Moore \u2013 Life form: Herb. Habitat: Epiphytic in moist montane forest, 1400\u20132800 m. Voucher: Someren 387 (EA).Elaphoglossumconforme (Sw.) Schott \u2013 Life form: Herb. Habitat: Moist montane forest, 2100\u20133000 m. Voucher: Balbo 769 (EA).Elaphoglossumdeckenii (Kuhn) C.Chr. \u2013 Life form: Herb. Habitat: Epiphytic in moist montane forest, 2100\u20133500 m. Voucher: Hedberg 1635 (EA).Elaphoglossumhybridum (Bory) Brack. \u2013 Life form: Herb. Habitat: Moist forest and damp places in the moorland, 1800\u20133600 m. Voucher: Faden et al. 74/1340 (EA).Elaphoglossumlastii (Baker) C.Chr. \u2013 Life form: Herb. Habitat: Epiphytic in moist montane forest, 1500\u20132600 m. Voucher: Faden et al. 74/1324 (EA).Elaphoglossumpiloselloides (C.Presl) T.Moore \u2013 Life form: Exotic herb. Habitat: Moist montane forest, 900\u20132450 m. Voucher: Peter 41097 (EA).*Elaphoglossumspatulatumvar.uluguruense (Reimers) Schelpe \u2013 Life form: Herb. Habitat: Moist forest and along streams, 900\u20132450 m. Voucher: Faden et al. 71/74 (EA).Elaphoglossumsubcinnamomeum (Christ) Hieron. \u2013 Life form: Herb. Habitat: Upper parts of wet montane forest, 2800\u20133600 m. Voucher: Kenya Exploration Society 157 (EA).Megalastrumlanuginosa (Willd. ex Kaulf.) Holttum \u2013 Life form: Herb. Habitat: Moist forest near streams, 1400\u20132400 m. Voucher: Faden et al. 71/282 (EA).Nothoperanemasquamisetum (Hook.) Ching \u2013 Life form: Herb. Habitat: Moist forest, 1850\u20132950 m. Voucher: Molesworth Allen 3638 (EA).Polystichumsinense (Christ) Christ \u2013 Life form: Herb. Habitat: Upland moist forest, 1920\u20134100 m. Voucher: Mwangangi 987 (EA).Polystichumtransvaalense N.C.Anthony \u2013 Life form: Herb. Habitat: Moist forest, 1350\u20132700 m. Voucher: Andrew 4461 (EA).Polystichumvolkensii (Hieron.) C.Chr. \u2013 Life form: Herb. Habitat: Moist montane forest and Hagenia forest, 2800\u20133600 m. Voucher: Rabb et al. 7 (EA).Polystichumwilsonii Christ \u2013 Life form: Herb. Habitat: Shaded grounds in moist forest, 2320\u20133650 m. Voucher: Someren 1053 (EA).EquisetaceaeF10. Equisetumramosissimum Desf. \u2013 Life form: Herb. Habitat: Along streams and rivers, 150\u20132100 m. Voucher: Greenway 13100 (EA).HymenophyllaceaeF11. Crepidomanesmelanotrichum (Schltdl.) J.P.Roux \u2013 Life form: Herb. Habitat: Shades in moist forest, 750\u20132650 m. Voucher: Beentje 2950 (EA).Crepidomanesramitrichum (Faden) Beentje \u2013 Life form: Herb. Habitat: Upland moist forest and rocky waterfalls, 2300\u20132600 m. Voucher: Faden & Grumbley 72/338 (EA).Didymoglossumerosum (Willd.) J.P.Roux \u2013 Life form: Herb. Habitat: Shady sites in moist forest, 0\u20132400 m. Voucher: Faden et al. 74/1336 (EA).Hymenophyllumcapillarevar.alternialatum (Pic.Serm.) Faden \u2013 Life form: herb. Habitat: Epiphytic in moist montane forest, 1650\u20133480 m. Voucher: Agnew et al. 5620 (EA).Hymenophyllumpolyanthosvar.kuhnii (C.Chr.) Schelpe \u2013 Life form: Herb. Habitat: Epiphytic in moist montane forest, 1400\u20133000 m. Voucher: Faden et al. 71/199 (EA).Hymenophyllumtunbrigense (L.) Sm. \u2013 Life form: Herb. Habitat: Epiphytic in upland moist forest, 1900\u20132700 m. Voucher: Faden et at. 74/1338 (EA).Polyphlebiumborbonicum (Bosch) Ebihara & Dubuisson \u2013 Life form: Herb. Habitat: Moist forest and stream banks, 1400\u20132600 m. Voucher: Faden et al. 71/196 (EA).MarsileaceaeF12. Marsileaminuta L. \u2013 Life form: Herb. Habitat: Aquatic in shallow water pools, edges of streams and seasonal swampy grassland, 0\u20131950 m. Voucher: Perkins 11502 (EA).OphioglossaceaeF13. Ophioglossumvulgatumsubsp.africanum Pocock ex J.E.Burrows \u2013 Life form: Herb. Habitat: Montane grassland, 1000\u20133250 m. Vouchers: Dale 1321, Faden & Faden 71/890 (EA).PolypodiaceaeF14. Drynariavolkensii Hieron. \u2013 Life form: Herb. Habitat: Riverine forest and woodland, 1600\u20132300 m. Voucher: Kerfoot 2767 (EA).Grammitiscryptophlebia (Baker) Copel. \u2013 Life form: Herb. Habitat: Epiphytic in moist montane forest, 1900\u20132150 m. Voucher: Faden 71/280 (EA).Lepisorusexcavatus (Bory ex Willd.) Ching \u2013 Life form: Herb. Habitat: Montane forest, 1150\u20133490 m. Voucher: Kamau 373 (EA).Lepisorusschraderi (Mett.) Ching \u2013 Life form: Herb. Habitat: Upland forest, 1200\u20132450 m. Voucher: Faden 74/904 (EA).Loxogrammeabyssinica (Baker) M.G.Price \u2013 Life form: Herb. Habitat: Epiphytic in moist forest, 900\u20132900 m. Voucher: Kirika et al. 70 (EA).Melpomeneflabelliformis (Poir.) A.R.Sm. & R.C.Moran \u2013 Life form: Herb. Habitat: Epiphytic in bushland, upland forest and bamboo zone, 1000\u20134200 m. Voucher: Fries 1325 (EA).Pleopeltismacrocarpa (Bory ex Willd.) Kaulf. \u2013 Life form: Herb. Habitat: Upper parts of montane forest, 1000\u20133600 m. Voucher: Vorontsova 55 (EA).PteridaceaeF15. Adiantumpoiretii Wikstr. \u2013 Life form: Herb. Habitat: Montane forest, 1000\u20132700 m. Voucher: Luke 1106 (EA).Adiantumraddianum C.Presl \u2013 Life form: Herb. Habitat: Wet rocky sites in montane forest, 1000\u20132700 m. Vouchers: Faden 68/765 & 68/849 (EA).Aleuritopterisfarinosa (Forsk.) F\u00e9e \u2013 Life form: Herb. Habitat: Swampy places in montane forest, 1460\u20133600 m. Voucher: Kamau 531 (EA).Cheilanthesbergiana Schltdl. \u2013 Life form: Herb. Habitat: Moist forest, 1300\u20132300 m. Voucher: Kamau 101 (EA).Cheilanthesquadripinnata (Forssk) Kuhn \u2013 Life form: Herb. Habitat: Rocky grounds in moist forest, 1275\u20132750 m. Voucher: Maas Geesteranus 5043 (EA).Coniogrammeafricana Hieron. \u2013 Life form: Herb. Habitat: Moist forest and along streams, 1000\u20132250 m. Voucher: Bytebier 3229 (EA).Oeosporangiumviride (Forssk.) Fraser-Jenk. & Pariyar \u2013 Life form: Herb. Habitat: Moist montane forest and shades in bushlands, 650\u20132250 m. Voucher: Faden 012/2003 (EA).Pteriscatoptera Kunze \u2013 Life form: Herb. Habitat: Wet or dry forest, 1000\u20133050 m. Voucher: Zogg 2547 (EA).Pterisdentata Forssk. \u2013 Life form: Herb. Habitat: Wet or dry forest, 1000\u20133000 m. Voucher: Kuchar 5203 (EA).TectariaceaeF16. Arthropterismonocarpa (Cordem.) C.Chr. \u2013 Life form: Herb. Habitat: Epiphytic in moist forest and riverine forest, 1250\u20132450 m. Voucher: Gillett and Holttum 20096 (EA).Tectariagemmifera (F\u00e9e) Alston \u2013 Life form: Herb. Habitat: Moist forest, 600\u20132550 m. Voucher: Strange 117 (EA).ThelypteridaceaeF17. Amauropeltaoppositiformis (C.Chr.) Holttum \u2013 Life form: Herb. Habitat: Wet or swampy sites in evergreen forest, 1200\u20133000 m. Voucher: Bytebier 222 (EA).Christelladentata (Forssk.) Brownsey & Jermy \u2013 Life form: Herb. Habitat: Along streams and damp places in forest, 45\u20132200 m. Voucher: Luke 148 (EA).Pneumatopterisunita (Kunze) Holttum \u2013 Life form: Herb. Habitat: Moist montane forest and bamboo thicket, 1450\u20132500 m. Voucher: Kamau 466 (EA).Pseudocyclosoruspulcher (Bory ex Willd.) Holttum \u2013 Life form: Herb. Habitat: Riverine forest and swampy sites in forest, 750\u20132250 m. Voucher: Faden 68/988 (EA).Phegopteriscruciata (Willd.) Mett. ex Kuhn \u2013 Life form: Herb. Habitat: Moist evergreen forest and stream banks, 1450\u20132350 m. Voucher: Faden & Evans 69/891 (EA).Stegnogrammapozoivar.petiolata (Ching) Sledge \u2013 Life form: Herb. Habitat: Wet montane forest, 2050\u20133350 m. Vouchers: Bytebier et al. 48, Faden & Faden 69/898 (EA).CupressaceaeF18. Cupressuslusitanica Mill. \u2013 Life form: Exotic tree. Habitat: Cultivated, 2600\u20132640 m. Voucher: Dyson 526 (EA).*Juniperusprocera Hochst. ex Endl. \u2013 Life form: Tree. Habitat: Upland dry evergreen forest, 1050\u20133250 m. Voucher: SK 0106 .PinaceaeF19. Pinuspatula Schiede ex Schltdl & Cham. \u2013 Life form: Exotic tree. Habitat: Cultivated, common in moist or dry forest, 1700\u20133000 m. Voucher: Althof s.n. (EA).*Pinusradiata D.Don \u2013 Life form: Tree. Habitat: Moist or dry forest, 1700\u20133000 m. Voucher: Dillon 4 (EA).PodocarpaceaeF20. Afrocarpusfalcatus (Thunb.) C.N.Page \u2013 Life form: Tree. Habitat: Dry evergreen forest, 1250\u20132700 m. Voucher: SK 0123 .Podocarpuslatifolius (Thunb.) R.Br. ex Mirb. \u2013 Life form: Tree. Habitat: Dry evergreen forest, 1500\u20133350 m. Voucher: SK 0102 .AlismataceaeF21. Alismaplantago-aquatica L. \u2013 Life form: Herb. Habitat: Marshes and stream banks, 900\u20132340 m. Voucher: Lubai 14 (EA).AmaryllidaceaeF22. Scadoxusmultiflorus (Martyn) Raf. \u2013 Life form: Herb. Habitat: Moist montane forest, 0\u20132700 m. Voucher: Mungai 1/83 (EA).AraceaeF23. Arisaemamildbraedii Engl. \u2013 Life form: Herb. Habitat: Wet and shaded places in montane forest, 1400\u20132620 m. Voucher: SK 0219 .Culcasiafalcifolia Engl. \u2013 Life form: Herbaceous climber. Habitat: Moist forest, 500\u20132100 m. Voucher: Luke 14154 (EA).Lemnaminor L. \u2013 Life form: Herb. Habitat: Surface of water pools and slow running streams, 0\u20131800 m. Voucher: Verdcourt 718b (EA).Zantedeschiapentlandii (R.Whyte ex W.Watson) Wittm. \u2013 Life form: Herb. Habitat: Along streams and swamps, 1400\u20131800 m. Voucher: SK 0255 .ArecaceaeF24. Phoenixreclinata Jacq. \u2013 Life form: Tree. Habitat: Open rocky slopes in rainforest, 0\u20133000 m. Voucher: Napier 5377 (EA).AsparagaceaeF25. Anthericumangustifolium Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland grassland, 1800\u20132850 m. Voucher: Leaky 8547 (EA).Asparagusafricanus Lam. \u2013 Life form: Woody climber. Habitat: Forest margins and wooded grassland, 0\u20133500 m. Voucher: SK 0191.Asparagusaridicola Sebsebe \u2013 Life form: Woody climber. Habitat: Wooded grassland and bush thickets, 10\u20132750 m. Voucher: Luke 10850 (EA).Asparagusasparagoides (L.) Druce \u2013 Life form: Herbaceous climber. Habitat: Moist forest and forest margins, 1100\u20133000 m. Voucher: Someren 1156 (EA).Asparagusfalcatus L. \u2013 Life form: Herbaceous climber. Habitat: Bush thickets and forest margins, 10\u20132750 m. Voucher: Zogg et al. 10/255 (EA).Asparagusnatalensis (Baker) J.-P.Lebrun & Stork \u2013 Life form: Woody climber. Habitat: Dry forest and forest margins, 900\u20132700 m. Voucher: Luke 18171 (EA).Asparagusracemosus Willd. \u2013 Life form: Woody climber. Habitat: Forest margins and wooded grassland, 1160\u20132900 m. Voucher: SK 0215 .Asparagussetaceus (Kunth) Jessop \u2013 Life form: Woody climber. Habitat: Forest margins, 1740\u20132300 m. Voucher: Verdcourt 3628 (EA).Chlorophytumcomosum (Thunb.) Jacques \u2013 Life form: Herb. Habitat: Undergrowth in rainforest, 20\u20132450 m. Voucher: SK 0168 .Chlorophytumpolystachys Baker \u2013 Life form: Herb. Habitat: Open woodland, 150\u20132900 m. Voucher: Hooper and Townsend 1651 (EA).Dracaenaafromontana Mildbr. \u2013 Life form: Tree. Habitat: Upland moist forest, 1600\u20132700 m. Voucher: SK 0143 .Dracaenaellenbeckiana Engl. \u2013 Life form: Tree. Habitat: Rocky slopes in moist forest, 1050\u20132100 m. Voucher: Perdue and Kibuwa 8259 (EA).Dracaenasteudneri Engl. \u2013 Life form: Tree. Habitat: Moist forest margins, 850\u20132300 m. Voucher: Perdue and Kibuwa 8023 (EA).Ornithogalumgracillimum R.E.Fr. \u2013 Life form: Herb. Habitat: Wet grounds in grassland and swamps, 1800\u20132700 m. Voucher: Polhill 406 (EA).Sansevieriaparva N.E.Br. \u2013 Life form: Herb. Habitat: Dry forest and rocky sites in bushland, 1600\u20132200 m. Voucher: Hansen 770 (EA).Sansevieria Perrotii Warb. \u2013 Life form: Herb. Habitat: Wooded grassland, 550\u20131950 m. Voucher: Someren 8505 (EA).ColchicaceaeF26. Androcymbiumstriatum Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland grassland and bushland, 1500\u20133400 m. Voucher: Rayner 66 (EA).Wurmbeatenuissubsp.hamiltonii (Wendelbo) B.Nord. \u2013 Life form: Herb. Habitat: Upland grassland, 2130\u20132750 m. Vouchers: Chandler 2415, Rayner 66 (EA).CommelinaceaeF27. Aneilemaleiocaule K.Schum. \u2013 Life form: Herb. Habitat: Moist forest mostly in shades, 1000\u20132740 m. Voucher: Dyson 570 (EA).Commelinaafricana L. \u2013 Life form: Herb. Habitat: Grassland and woodland, 300\u20132980 m. Voucher: Kerfoot 619 (EA).Commelinaimberbis Ehrenb. ex Hassk. \u2013 Life form: Herb. Habitat: Grassland and bushland, 1000\u20132910 m. Voucher: SK 0166 .Commelinalatifolia Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland grassland, 1250\u20132270 m. Voucher: Kenya Forest excursion 26 (EA).Commelinareptans Brenan \u2013 Life form: Herb. Habitat: Moist grassland, 1200\u20132550 m. Voucher: Faden 71/889 (EA).Floscopaglomerata (Willd. ex Schult. & Schult.f.) Hassk. \u2013 Life form: Herb. Habitat: Swampy grassland and along streams, 900\u20132200 m. Voucher: Bally 13225 (EA).Murdanniaclarkeana Brenan \u2013 Life form: Herb. Habitat: Swampy grassland, 1500\u20131850 m. Voucher: Hooper et al. 1695 (EA).Murdanniasimplex (Vahl) Brenan \u2013 Life form: Herb. Habitat: Grassland and bushland, 30\u20132200 m. Voucher: Verdcourt 561 (EA).CyperaceaeF28. Bulbostylisglaberrima K\u00fck. \u2013 Life form: Herb. Habitat: Damp sites in moorland, 3000\u20133600 m. Voucher: Fries and Fries 2394 (EA).Carexbequaertii De Wild. \u2013 Life form: Herb. Habitat: Moist montane forest and bamboo thickets, 1950\u20133800 m. Voucher: Verdcourt 1769 (EA).Carexchlorosaccus C.B.Clarke \u2013 Life form: Herb. Habitat: Moist forest and riparian forest, 1300\u20133300 m. Voucher: Napper 715 (EA).Carexconferta Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland moist forest and moorland, 2200\u20133650 m. Voucher: Robertson 7373(EA).Carexelgonensis Nelmes \u2013 Life form: Herb. Habitat: Bamboo thicket margins and afro-alpine stream banks, 2400\u20133650 m. Voucher: Hedberg 854 (EA).Carexjohnstonii Boeckeler \u2013 Life form: Herb. Habitat: Upper parts of montane forest and bamboo zone, 2200\u20133300 m. Voucher: Musili et al. 422 (EA).Carexlycurus K.Schum. \u2013 Life form: Herb. Habitat: Stream banks in grassland and woodland, 1500\u20133350 m. Voucher: Verdcourt 1770 (EA).Carexmonostachya A.Rich. \u2013 Life form: Herb. Habitat: Upper regions of bamboo zone and moorland, 2700\u20134500 m. Voucher: Musili et al 439 (EA).Carexperegrina Link \u2013 Life form: Herb. Habitat: Moist montane forest, 2300\u20133440 m. Voucher: Muasya et al. 050 (EA).Carexphragmitoides K\u00fck. \u2013 Life form: Herb. Habitat: Bogs and marshes in montane forest, 2500\u20133100 m. Voucher: Taylor 1354 (EA).ECarexrunssoroensisvar.aberdarensis K\u00fck. \u2013 Life form: Herb. Habitat: Rocky grounds in moorland, 3000\u20134400 m. Vouchers: Hedberg 4327, Muasya et al. 048 (EA).Carexsimensis Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Swampy places in upland grassland and moorland, 1850\u20133900 m. Voucher: Brich 61/13 (EA).Carexvallis-rosetto K.Schum. \u2013 Life form: Herb. Habitat: Moist sites in forest and forest edges, 1000\u20133300 m. Voucher: Luke 15347 (EA).Cyperusafroalpinus Lye \u2013 Life form: Herb. Habitat: Open sites in montane forest and bamboo thickets, 1000\u20133000 m. Voucher: Haines 1969 (EA).Cyperusajax C.B.Clarke \u2013 Life form: Herb. Habitat: Roadsides in upland forest and bush thickets, 950\u20132600 m. Voucher: Napper 1826 (EA).Cyperusaterrimus Hochst. ex Steud. \u2013 Life form: Herb. Habitat: Damp sites in upland montane forest, 1000\u20133350 m. Voucher: Brown 358 (EA).Cyperuscyperoides (L.) Kuntze \u2013 Life form: Herb. Habitat: Roadsides and forest clearings, 600\u20132400 m. Voucher: Kibui 43 (EA).Cyperusdenudatus L.f. \u2013 Life form: Herb. Habitat: Damp grassland and riversides, 0\u20132000 m. Voucher: Kuchar 7820 (EA).Cyperusdereilema Steud. \u2013 Life form: Herb. Habitat: Moist montane forest and bamboo zone, 2100\u20133050 m. Voucher: Luke 15351 (EA).Cyperusdichroostachyus Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Moist forest, 1200\u20132750 m. Voucher: Robertson 7372 (EA).Cyperusesculentus L. \u2013 Life form: Herb. Habitat: Swamps and wet grassland, 0\u20132200 m. Voucher: Faden 68/854 (EA).Cyperuskarisimbiensis (Cherm.) K\u00fck. \u2013 Life form: Herb. Habitat: Woodland, 1850\u20133000 m. Voucher: Fries and Fries 1037 (EA).Cyperuskerstenii Boeckeler \u2013 Life form: Herb. Habitat: Montane grassland and moorland, 2400\u20133600 m. Voucher: Beentje et al. 62 (EA).Cyperuspapyrus L. \u2013 Life form: Herb. Habitat: Moist forest and wet grassy slopes, 1800\u20133120 m. Voucher: Wood 779 (EA).Cyperusrigidifolius Steud. \u2013 Life form: Herb. Habitat: Seasonally wet grassland and bushland, 1700\u20132800 m. Voucher: Robertson 7399 (EA).Cyperusrotundus L. \u2013 Life form: Herb. Habitat: Wet grassland, 0\u20132200 m. Voucher: Robertson 7368 (EA).Cyperustomaiophyllus K.Schum. \u2013 Life form: Herb. Habitat: Moist forest and damp grassy slopes, 1800\u20133120 m. Voucher: Kuchar 12533 (EA).Eleocharismarginulata Hochst. ex Steud. \u2013 Life form: Herb. Habitat: Swampy grassland, 1500\u20132600 m. Voucher: Robertson 7369 (EA).Fimbristyliscomplanatasubsp.keniaeensis (K\u00fck.) Lye. \u2013 Life form: Herb. Habitat: Swampy grassland, 1500\u20132700 m. Vouchers: Faden 67/771, Kabuye 378 (EA).Fimbristylisovata (Burm.f.) J.Kern \u2013 Life form: Herb. Habitat: Wet wooded grassland, 0\u20132200 m. Voucher: Verdcourt 3253 (EA).Fuirenapubescens (Poir) Kunth \u2013 Life form: Herb. Habitat: Seasonally wet grassland, 850\u20132300 m. Voucher: Musili et al. 191(EA).Isolepiscostata Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Moist montane forest and stream banks, 1700\u20133500 m. Voucher: Taylor 1274 (EA).Isolepisfluitans (L.) R.Br. \u2013 Life form: Herb. Habitat: Bogs in moorland, 1200\u20133700 m. Voucher: Okwaro 34 (EA).Isolepissepulcralis Steud. \u2013 Life form: Herb. Habitat: Upland wet grassland, 1800\u20132300 m. Voucher: Bogdan 1514a (EA).Isolepissetaceae (L.) R.Br. \u2013 Life form: Herb. Habitat: Wet montane grassland, 2400\u20133800 m. Voucher: Kuchar 8278 (EA).Kyllingabrevifoliavar.lurida (K\u00fck.) Beentje \u2013 Life form: Herb. Habitat: Montane grassland and forest clearings, 1300\u20133300 m. Voucher: Hansen 754 (EA).Cyperuserecta (Schumach.) Mattf. & K\u00fck. \u2013 Life form: Herb. Habitat: Wet depressions and swamps, 0\u20132000 m. Voucher: Musili et al. 244 (EA).Kyllingaodorata Vahl. \u2013 Life form: Herb. Habitat: Upland forest margins and woodland, 1300\u20133300 m. Voucher: Robertson 7370 (EA).Cyperusaethiops Welw. ex Ridl. \u2013 Life form: Herb. Habitat: Wet or swampy grassland, 900\u20132200 m. Voucher: Agnew et al. 8608 (EA).Cyperuselegantulus Steud. \u2013 Life form: Herb. Habitat: Wet grassland and moist forest margins, 1100\u20133050 m. Voucher: Napper 1488 (EA).Cyperusmundiivar.uniceps (C.B.Clarke) K\u00fck. \u2013 Life form: Herb. Habitat: Wet grassland and swampy forest, 200\u20132300 m. Voucher: Verdcourt 427 (EA).Cyperusnigricans Steud. \u2013 Life form: Herb. Habitat: Marshy grounds and bogs in upland forest, 1700\u20133600 m. Voucher: Kuchar 9559 (EA).Cyperusnitidus Lam. \u2013 Life form: Herb. Habitat: Wetland and swamps edges, 1000\u20132150 m. Voucher: Bogdan 2894 (EA).Carexuhligii K.Schum ex C.B.Clarke \u2013 Life form: Herb. Habitat: Open sites in moist forest and upland grassland, 1050\u20132800 m. Voucher: Hansen 850 (EA).Carexspartea Wahlenb. \u2013 Life form: Herb. Habitat: Moist forest edges and wet upland grassland, 1650\u20132800 m. Voucher: Polhill 436 (EA).EriocaulaceaeF29. Eriocaulonmesanthemoides Ruhland \u2013 Life form: Herb. Habitat: Stream-sides in montane grassland and moorland, 2400\u20133100 m. Voucher: Agnew et al. 8157 (EA).Eriocaulonschimperi K\u00f6rn. ex Ruhland \u2013 Life form: Herb. Habitat: Wet montane grassland, 2000\u20133500 m. Voucher: Wood 773 (EA).Eriocaulonvolkensii Engl. \u2013 Life form: Herb. Habitat: Montane grassland and moorland, 2500\u20133900 m. Voucher: Agnew 7226 (EA).HydrocharitaceaeF30. Egeriadensa Planch. \u2013 Life form: Herb. Habitat: Still or slow flowing freshwater, 360\u20132400 m. Voucher: Powys 373 (EA).HypoxidaceaeF31. Hypoxisangustifolia Lam. \u2013 Life form: Herb. Habitat: Open woodland and forest margins, 0\u20133000 m. Voucher: Verdcourt 1965 (EA).Hypoxiskilimanjaricasubsp.prostrata Ellen Holt & Staubo \u2013 Life form: Herb. Habitat: Montane grassland, 2900\u20133500 m. Vouchers: Mabberlay 372, Beentje 2611 (EA).IridaceaeF32. Aristeaalata Baker \u2013 Life form: Herb. Habitat: Moist forest and forest edges, 1800\u20133500 m. Voucher: Mbale et al. 850 (EA).Aristeaangolensis Baker \u2013 Life form: Herb. Habitat: Wet sites in grassland, 1750\u20132700 m. Voucher: Battiscombe 831 (EA).Dieramacupuliflorum Klatt \u2013 Life form: Herb. Habitat: Montane grassland, 2000\u20133900 m. Vouchers: Kuchar 12699, Beentje 2414 (EA).Gladiolusdalenii Van Geel \u2013 Life form: Herb. Habitat: Grassland with scattered trees, 300\u20133600 m. Voucher: Bally 8255B (EA).Gladioluswatsonioides Baker \u2013 Life form: Herb. Habitat: Forest clearings in montane forest, 2000\u20133800 m. Voucher: Lind 2901 (EA).Hesperanthapetitiana (A.Rich.) Baker \u2013 Life form: Herb. Habitat: Rocky cliffs at subalpine grassland, 1800\u20133100 m. Voucher: Napier 1254 (EA).Romuleacongoensis B\u00e9g. \u2013 Life form: Herb. Habitat: Montane grassland, 3300\u20134200 m. Voucher: Stephenson 383 (EA).Romuleafischeri Pax \u2013 Life form: Herb. Habitat: Upland grassland, 2150\u20134200 m. Voucher: Dyson 435 (EA).JuncaceaeF33. Juncusbufonius L. \u2013 Life form: Herb. Habitat: Wet places in upland grassland, 2400\u20132800 m. Voucher: Musili et al. 441 (EA).Juncusdregeanussubsp.bachitii (Hochst. ex Steud.) Hedberg. \u2013 Life form: Herb. Habitat: Along streams in upland forest and moorland, 2100\u20133400 m. Vouchers: Hedberg 4328, Kuchar 12445 (EA).Juncuseffusus L. \u2013 Life form: Herb. Habitat: Moist forest and streams banks, 1500\u20133300 m. Voucher: Kuchar 10330 (EA).Juncusoxycarpus E.Mey ex Kunth \u2013 Life form: Herb. Habitat: Marshy sites in grassland, 1500\u20133080 m. Voucher: Handa 007 (EA).Luzulaabyssinica Parl. \u2013 Life form: Herb. Habitat: Damp sites in upper montane forest and moorland, 2000\u20134550 m. Voucher: Hedberg 1541 (EA).Luzulajohnstonii Buchenau \u2013 Life form: Herb. Habitat: Shaded grounds in rainforest, 2400\u20134200 m. Voucher: Agnew et al. 7088 (EA).OrchidaceaeF34. Aerangisconfusa J.Stewart \u2013 Life form: Herb. Habitat: Epiphytic in shaded trunks and branches in upland forest, 1600\u20132500 m. Voucher: Bally 8461 (EA).Aerangisthomsonii (Rolfe) Schltr. \u2013 Life form: Herb. Habitat: Epiphytic in shaded trunks and branches in upland forest, 1600\u20132600 m. Voucher: SK 0260 .Angraecumchamaeanthus Schltr. \u2013 Life form: Herb. Habitat: Epiphytic in upland montane forest, 1600\u20132400 m. Voucher: Smart 27 (EA).Angraecumconchiferum Lindl. \u2013 Life form: Herb. Habitat: Epiphytic in montane forest, 1250\u20132400 m. Voucher: Archer 20/9/1959 (EA).Angraecumhumile Summerh. \u2013 Life form: Herb. Habitat: Epiphytic in moist montane forest and riparian forest, 1650\u20132500 m. Voucher: Someren 413 (EA).Angraecumsacciferum Lindl. \u2013 Life form: Herb. Habitat: Epiphytic in upland moist forest, 900\u20132200 m. Voucher: Bytebier 505 (EA).Bulbophyllumsandersonii (Hook.f.) Rchb.f. \u2013 Life form: Herb. Habitat: Epiphytic in shades in forest, 200\u20132700 m. Voucher: Cameron 14863 (EA).Calanthesylvatica (Thouars) Lindl. \u2013 Life form: Herb. Habitat: Moist forest floors and near streams, 900\u20133000 m. Voucher: Gardner 1411 (EA).Cynorkisanacamptoides Kraenzl. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 1050\u20133350 m. Voucher: Verdcourt 434 (EA).Cyrtorchisarcuata (Lindl.) Schltr. \u2013 Life form: Herb. Habitat: Epiphytic in woodland and open forest, 0\u20133300 m. Vouchers: Gardner 2554, Padwa 59 (EA).Diaphanantherohrii (Rchb.f.) Summerh. \u2013 Life form: Herb. Habitat: Epiphytic in montane forest, 2100\u20133000 m. Voucher: Smart 22 (EA).Disafragranssubsp.deckenii (Rchb.f.) H.P.Linder \u2013 Life form: Herb. Habitat: Open sites and edges of montane forest up to the moorland, 2350\u20133700 m. Vouchers: Kokwaro 1929, Agnew et al. 8186 (EA).Disastairsii Kraenzl. \u2013 Life form: Herb. Habitat: Upland grassy swamps and damp sites in moorland, 2100\u20133750 m. Voucher: Kokwaro 3249 (EA).Disperisdicerochila Summerh. \u2013 Life form: Herb. Habitat: Upland moist forest, 1650\u20132600 m. Voucher: Williams 15379 (EA).Disperiskilimanjarica Rendle \u2013 Life form: Herb. Habitat: Shaded grounds in evergreen forest, 2100\u20133000 m. Voucher: SK 0133 .Epipactisafricana Rendle \u2013 Life form: Herb. Habitat: Montane evergreen forest and bamboo thickets, 2330\u20133750 m. Voucher: Verdcourt et al. 3022 (EA).Habenariaattenuata Hook.f. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 2700\u20134000 m. Voucher: Davis 7A (EA).Habenariadecorata Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Rocky sites in moorland, 2200\u20133300 m. Voucher: Ward s.n (EA).Habenariakeniensis Summerh. \u2013 Life form: Herb. Habitat: Upland rainforest, 1950\u20132950 m. Voucher: Belcher 151/54 (EA).Habenariamacrantha Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 2550\u20133000 m. Voucher: Dale 2842 (EA).Habenariapetitiana (A.Rich) T.Durand & Schinz \u2013 Life form: Herb. Habitat: Forest edges and bushland, 1500\u20133300 m. Voucher: William 15380 (EA).Habenariaschimperiana Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Swamps and wet grassland, 1250\u20132800 m. Voucher: Dale 2876 (EA).Habenariatweedieae Summerh. \u2013 Life form: Herb. Habitat: In grass on rocky hills, 1950\u20132600 m. Voucher: Cunningham 44 (EA).Habenariavaginata A.Rich. \u2013 Life form: Herb. Habitat: Wet sites in grassland, 1300\u20133000 m. Voucher: Dale 2354 (EA).Holothrixbrongniartiana Rchb.f. \u2013 Life form: Herb. Habitat: Upland grassland, 1200\u20133500 m. Voucher: Williams 12362 (EA).Liparisdeistelii Schltr. \u2013 Life form: Herb. Habitat: Epiphytic on fallen trees in shades near rivers, 1700\u20132750 m. Voucher: Dale 2861 (EA).Polystachyacaespitificasubsp.latilabris (Summerh.) P.J.Cribb & Podz. \u2013 Life form: Herb. Habitat: Epiphytic in montane forest, 1800\u20132200 m. Voucher: Lucs et al. 274 (EA).Polystachyacultriformis (Thouars) Lindl. ex Spreng \u2013 Life form: Herb. Habitat: Moist montane forest, 1800\u20132200 m. Vouchers: SK 0140, SK 00227 .Polystachyaheckmanniana Kraenzl. \u2013 Life form: Herb. Habitat: Upland moist forest, 1200\u20132000 m. Voucher: Turner 3231 (EA).Polystachyatransvaalensis Schltr. \u2013 Life form: Herb. Habitat: Moist forest, 1200\u20132900 m. Vouchers: Turner 3239 & 2385 (EA).Satyriumcrassicaule Rendle \u2013 Life form: Herb. Habitat: Wet grassland and swamps, 1000\u20133150 m. Voucher: Bytebier 500 (EA).Satyriummacrophyllum Lindl. \u2013 Life form: Herb. Habitat: Upland wet grassland, 1200\u20133150 m. Voucher: Dale 75 (EA).Satyriumschimperi Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland grassland, 2100\u20133200 m. Voucher: Pierce 2696 (EA).Satyriumvolkensii Schltr. \u2013 Life form: Herb. Habitat: Upland grassland and bushland, 1050\u20132400 m. Voucher: Piers 143/51 (EA).PoaceaeF35. Acritochaetevolkensii Pilg. \u2013 Life form: Herb. Habitat: Shaded grounds in upland forest and bamboo thicket, 2300\u20133300 m. Voucher: Agnew et al. 8180 (EA).Agrostisgracilifolia C.E.Hubb. \u2013 Life form: Herb. Habitat: Wet places in upland grassland and moorland, 2800\u20133980 m. Voucher: Beentje 3269(EA).Agrostiskeniensis Pilg. \u2013 Life form: Herb. Habitat: Along streams and moist forest margins, 2200\u20133000 m. Voucher: Turner 1569 (EA).Agrostiskilimandscharica Mez \u2013 Life form: Herb. Habitat: Upland forest clearings and margins up to bamboo zone, 2000\u20134000 m. Voucher: Agnew et al. 8134 (EA).Agrostisproducta Pilg. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 2400\u20134000 m. Voucher: Robertson 2162 (EA).Agrostistrachyphylla Pilg. \u2013 Life form: Herb. Habitat: Wet places in upland grassland and moorland, 3500\u20134900 m. Voucher: AfroAlp II team 1063 (EA).Agrostisvolkensii Stapf \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 3000\u20133900 m. Voucher: AfroAlp II team 0974 (EA).Airacaryophyllea L. \u2013 Life form: Herb. Habitat: Rocky soils in upland grassland and moorland, 2000\u20134500 m. Voucher: Rauh 407 (EA).Andropogonabyssinicus R.Br. ex Fresen. \u2013 Life form: Herb. Habitat: Upland grassland, 200\u20133100 m. Voucher: Kuchar 12501 (EA).Andropogonamethystinus Steud. \u2013 Life form: Herb. Habitat: Open or clearings in montane forest and moorland, 1400\u20134000 m. Voucher: Grant 1238 (EA).Andropogonchrysostachyus Steud. \u2013 Life form: Herb. Habitat: Moist upland grassland and evergreen forest, 2100\u20133300 m. Voucher: Napper 745 (EA).Andropogondistachyos L. \u2013 Life form: Herb. Habitat: Upland dry grassland, 1700\u20133000 m. Voucher: Kuchar 12321 (EA).Andropogonlima (Hack.) Stapf \u2013 Life form: Herb. Habitat: Montane grassland and dry moorland, 2400\u20134000 m. Voucher: Rauh 522 (EA).Anthoxanthumnivale K.Schum. \u2013 Life form: Herb. Habitat: Upland moist grassland and moorland, 2500\u20133980 m. Voucher: Kuchar 12739(EA).Aristidaadoensis Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Deciduous bushland, 1300\u20132300 m. Voucher: Robertson 332 (EA).Aristidajunciformis Trin. & Rupr. \u2013 Life form: Herb. Habitat: Dry rocky hilltops, 400\u20132100 m. Voucher: Bogdan 4163 (EA).Avenafatua L. \u2013 Life form: Herb. Habitat: Upland grassland, 2100\u20132400 m. Voucher: Ghosh 4 (EA).Avenasterilis L. \u2013 Life form: Herb. Habitat: Upland grassland, 2600\u20132600 m. Voucher: Someren 13/12/1956 (EA).Bothriochloainsculpta (A.Rich.) A.Camus \u2013 Life form: Herb. Habitat: Grassland, 0\u20132250 m. Voucher: Hansen 800 (EA).Brachypodiumflexum Nees \u2013 Life form: Herb. Habitat: Moist forest and bamboo thickets, 2000\u20133000 m. Voucher: Kuchar and Msafiri 5422 (EA).Brizamaxima L. \u2013 Life form: Herb. Habitat: Roadsides in forest, 2400\u20132700 m. Voucher: Edwards 2833/6 (EA).Bromuscatharticus Vahl \u2013 Life form: Herb. Habitat: Disturbed grounds and roadsides, 2300\u20132700 m. Vouchers: Greenway 10399 & 9574 (EA).Bromusdiandrus Roth \u2013 Life form: Herb. Habitat: Disturbed grounds and roadsides, 2300\u20133000 m. Voucher: Muchiri 580 (EA).Bromusleptoclados Nees \u2013 Life form: Herb. Habitat: Upland grassland and forest clearings, 2300\u20134300 m. Voucher: Bogdan 2646 (EA).Calamagrostisepigejos (L.) Roth \u2013 Life form: Herb. Habitat: Upland grassland and forest clearings, 2000\u20133000 m. Voucher: Kokwaro 3340 (EA).Calamagrostishedbergii Melderis \u2013 Life form: Herb. Habitat: Rocky moorland, 3500\u20134250 m. Voucher: Hedberg 1810 (EA).Chlorisvirgata Sw. \u2013 Life form: Herb. Habitat: Scattered-tree grassland and bushland, 10\u20132120 m. Voucher: Robertson 2218 (EA).Coelachnefriesiorum C.E.Hubb. \u2013 Life form: Herb. Habitat: Moist montane forest, 3000\u20133000 m. Voucher: Fries 2407 (EA).Colpodiumhedbergii (Melderis) Tzvelev \u2013 Life form: Herb. Habitat: Moorland, 3580\u20134000 m. Voucher: Hanid 157 (EA).Cymbopogonnardus (L.) Rendle \u2013 Life form: Herb. Habitat: Upland grassland, 1000\u20133000 m. Voucher: Waiganjo 39 (EA).Cynodondactylon (L.) Pers. \u2013 Life form: Herb. Habitat: Roadsides in grassland, 0\u20132000 m. Voucher: Mbevi 194 (EA).Deschampsiacespitosa (L.) P.Beauv. \u2013 Life form: Herb. Habitat: Damp places in moorland, 2900\u20134000 m. Voucher: Polhill 12035 (EA).Deschampsiaflexuosa (L.) Trin. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 2600\u20133920 m. Voucher: Agnew et al. 8135A (EA).Digitariaabyssinica (A.Rich.) Stapf \u2013 Life form: Herb. Habitat: Ruderal sites in upland grassland, 0\u20133000 m. Voucher: Turner 1564 (EA).Digitariadiagonalis (Nees) Stapf \u2013 Life form: Herb. Habitat: Grassland, 0\u20132640 m. Voucher: Mbale 1024 (EA).Digitariagazensis Rendle \u2013 Life form: Herb. Habitat: Upland grassland, 1200\u20132600 m. Voucher: Barney 1031 (EA).Digitarialongiflora (Retz.) Pers. \u2013 Life form: Herb. Habitat: Deciduous bushland, 0\u20132300 m. Voucher: Robertson 2131 (EA).Digitariathouaresiana (Fl\u00fcgg\u00e9) A.Camus \u2013 Life form: Herb. Habitat: Moist forest, 0\u20132400 m. Voucher: Kerfoot 1359 (EA).Digitariavelutina (Forssk.) P.Beauv. \u2013 Life form: Herb. Habitat: Roadsides and other disturbed places, 0\u20132300 m. Voucher: Robertson 7337 (EA).Ehrhartaerecta Lam. \u2013 Life form: Herb. Habitat: Upland forest glades and margins, 1500\u20132700 m. Voucher: Robertson 7350 (EA).Eleusinejaegeri Pilg. \u2013 Life form: Herb. Habitat: Upland grassland and open sites in forest, 1800\u20133300 m. Voucher: Turner 1573 (EA).EEragrostisamanda Clayton \u2013 Life form: Herb. Habitat: Glades in bamboo thickets, 2400\u20132900 m. Voucher: Dyson 442 (EA).Eragrostischalarothyrsos C.E.Hubb. \u2013 Life form: Herb. Habitat: Swampy grassland, 1100\u20132500 m. Voucher: Lepelley 5 (EA).Eragrostisolivacea K.Schum. \u2013 Life form: Herb. Habitat: Rocky sites in grassland, 1300\u20133300 m. Voucher: Napper 1699 (EA).Eragrostispatula (Kunth) Steud. \u2013 Life form: Herb. Habitat: Roadsides and disturbed sites, 0\u20132800 m. Voucher: Verdcourt 3642 (EA).Eragrostisschweinfurthii Chiov. \u2013 Life form: Herb. Habitat: Upland evergreen forest, 1300\u20133000 m. Voucher: Beentje 3234 (EA).Eragrostistef (Zucc.) Trotter \u2013 Life form: Herb. Habitat: Roadsides and disturbed areas, 750\u20132500 m. Voucher: Frazer 195 (EA).Exothecaabyssinica (Hochst. ex A.Rich.) Andersson \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 2000\u20134000 m. Voucher: Coe 771 (EA).Festucaafricana (Hack.) Clayton \u2013 Life form: Herb. Habitat: Shaded sites in upland forest and bamboo thickets, 2000\u20133000 m. Voucher: Bogdan 2832 (EA).Festucaarundinacea Schreb. \u2013 Life form: Herb. Habitat: Along stream banks in upland forests, 2300\u20133090 m. Voucher: Bogdan 4760 (EA).Festucacamusiana St.-Yves \u2013 Life form: Herb. Habitat: Upland forest and bamboo thicket, 2100\u20133500 m. Voucher: Kerfoot 1428 (EA).Festucacostata Nees \u2013 Life form: Herb. Habitat: Upland grassland, 2400\u20133000 m. Voucher: Schelpe 2684 (EA).Festucamekiste Clayton \u2013 Life form: Herb. Habitat: Forest edges and open sites in upland forest, 2300\u20133480 m. Voucher: Bogdan 3274 (EA).Festucapilgeri St.-Yves \u2013 Life form: Herb. Habitat: Moorland, 2700\u20134250 m. Voucher: Agnew and Timberlake 11165 (EA).Festucasimensis Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Shaded places in upland forest, 2000\u20133300 m. Voucher: Robertson 3935 (EA).Harpachneschimperi A.Rich. \u2013 Life form: Herb. Habitat: Grassland and open bushland, 500\u20133000 m. Voucher: Faden et al. 74/613 (EA).Helictotrichonelongatum (Hochst. ex A.Rich.) C.E.Hubb. \u2013 Life form: Herb. Habitat: Upland forest edges and grassland, 1800\u20133800 m. Voucher: Kuchar and Msafiri 5407 (EA).Helictotrichonmilanjianum (Rendle) C.E.Hubb. \u2013 Life form: Herb. Habitat: Moist shaded places in montane forest and bamboo thicket, 2300\u20133500 m. Voucher: Kerfoot 1424 (EA).Helictotrichonumbrosum (Hochst. ex Steud.) C.E.Hubb. \u2013 Life form: Herb. Habitat: Upland grassland and margins of bamboo thickets, 1850\u20134000 m. Voucher: Peacock 58 (EA).Hyparrheniamobukensis (Chiov.) Chiov. \u2013 Life form: Herb. Habitat: Margins of montane evergreen forest and bamboo forest, 2500\u20133300 m. Voucher: Kerfoot 1353 (EA).Hyparrheniatamba (Hochst. ex Steud.) Andersson ex Stapf \u2013 Life form: Herb. Habitat: Upland grassland, 2000\u20133300 m. Voucher: Kerfoot 431 (EA).Hyparrheniaumbrosa (Hochst.) Andersson ex Clayton \u2013 Life form: Herb. Habitat: Roadsides in lower montane forest, 1500\u20132300 m. Voucher: Glover et al. 1424 (EA).Koeleriacapensis Nees \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 1800\u20135300 m. Voucher: Grout 1247 (EA).Leersiadenudata Launert \u2013 Life form: Herb. Habitat: Swampy grassland, 1500\u20132300 m. Voucher: Faden 74/671(EA).Loliumtemulentum L. \u2013 Life form: Herb. Habitat: Grazed pasture in moist forest, 1900\u20132300 m. Voucher: Lundin 5097 (EA).Oplismenushirtellus (L.) P.Beauv. \u2013 Life form: Herb. Habitat: Shaded grounds in forest, 0\u20132500 m. Voucher: Nattrass 716 (EA).Oplismenusundulatifolius (Ard.) Roem & Schult. \u2013 Life form: Herb. Habitat: Shaded grounds in forest, 1400\u20132500 m. Voucher: Bogdan 378 (EA).Panicumcalvum Stapf \u2013 Life form: Herb. Habitat: Shades in forest and forest margins, 1000\u20133000 m. Voucher: Davidse 7050 (EA).Panicumhymeniochilum Nees \u2013 Life form: Herb. Habitat: River banks and swamps, 700\u20133120 m. Voucher: Kabuye & Wood 100 (EA).Panicummonticola Hook.f. \u2013 Life form: Herb. Habitat: Shaded forest floors, 600\u20132600 m. Voucher: Napper et al. 1719 (EA).Panicumpusillum Hook.f. \u2013 Life form: Herb. Habitat: Montane grassland or bushland, 1300\u20133300 m. Voucher: Bogdan 4881 (EA).Panicumsubalbidum Kunth \u2013 Life form: Herb. Habitat: Along rivers and swamps, 200\u20133400 m. Voucher: Kuchar 7835 (EA).Paspalumnotatum Fl\u00fcgg\u00e9 \u2013 Life form: Exotic herb. Habitat: Grassland, 500\u20132100 m. Voucher: Hindorf 693 (EA).*Pennisetumclandestinum Hochst. ex Chiov. \u2013 Life form: Herb. Habitat: Upland grassland, 1400\u20133300 m. Voucher: Mungai 65 (EA).Pennisetumhohenackeri Hochst. ex Steud. \u2013 Life form: Herb. Habitat: Upland grassland, 1100\u20132400 m. Voucher: Robertson 7322 (EA).Pennisetumriparium Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Swamps, 1400\u20132700 m. Voucher: Bogdan 3514 (EA).Pennisetumsphacelatum (Nees) T.Durand & Schinz \u2013 Life form: Herb. Habitat: Upland grassland and evergreen forests, 1500\u20133200 m. Voucher: Bally 1177 (EA).Pennisetumthunbergii Kunth \u2013 Life form: Herb. Habitat: Upland pasture and roadsides in moist forest, 1500\u20133500 m. Voucher: Kerfoot 1422 (EA).Pennisetumtrachyphyllum Pilg. \u2013 Life form: Herb. Habitat: Moist forest often along paths and glades, 1000\u20132500 m. Voucher: Napper 1718 (EA).Pentamerisborussica (K.Schum.) Galley & H.P.Linder \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 3000\u20134680 m. Voucher: Kuchar 12524 (EA).Pentamerispictigluma (Steud.) Galley & H.P.Linder \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 2600\u20134500 m. Voucher: Hedberg 1538 (EA).Phalarisarundinacea L. \u2013 Life form: Herb. Habitat: Stream banks and swamps margins, 1950\u20133000 m. Voucher: Verdcourt 876 (EA).Poaannua L. \u2013 Life form: Herb. Habitat: Disturbed grounds in upland forest, 2000\u20133550 m. Voucher: Vorontsova 767 (EA).Poaleptoclada Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland forest margins and grassland, 1800\u20134750 m. Voucher: Vorontsova 768 (EA).Poaschimperiana Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 1980\u20134200 m. Voucher: Hedberg 1595 (EA).Poecilostachysoplismenoides (Hack.) Clayton \u2013 Life form: Herb. Habitat: Shaded places in evergreen forest, 1000\u20132500 m. Voucher: Bogdan 1504 (EA).Polypogonschimperianus (Hochst. ex Steud.) Cope \u2013 Life form: Herb. Habitat: Along rivers and moist places in upland grassland and moorland, 1500\u20134000 m. Voucher: Greenway 10400 (EA).Pseudechinolaenapolystachya Stapf \u2013 Life form: Herb. Habitat: Upland moist forest often in shady grounds, 1000\u20132500 m. Voucher: Faden 67374 (EA).Setariaatrata Hack. ex Engl. \u2013 Life form: Herb. Habitat: Swamps, 2000\u20132600 m. Voucher: Bogdan 2787 (EA).Setariamegaphylla (Steud.) T.Durand & Schinz \u2013 Life form: Herb. Habitat: Shaded places in forest, 200\u20132350 m. Voucher: SK 0246 .Setariasphacelata (Schumach.) Stapf & C.E.Hubb. ex Moss \u2013 Life form: Herb. Habitat: Wooded grassland, 0\u20133300 m. Voucher: Robertson 7312 (EA).Snowdeniapolystachya (Fresen.) Pilg. \u2013 Life form: Herb. Habitat: Upland grassland, 2100\u20132700 m. Voucher: Luke et al. 16120 (EA).Sporobolusafricanus (Poir.) Robyns & Tournay \u2013 Life form: Herb. Habitat: Grazed grassland, 1300\u20132640 m. Voucher: Robertson 7312 (EA).Sporobolusagrostoides Chiov. \u2013 Life form: Herb. Habitat: Shades in upland evergreen forest, 1300\u20132800 m. Voucher: Robertson 7348 (EA).Sporobolusfimbriatus (Trin.) Nees \u2013 Life form: Herb. Habitat: Open deciduous bushland, 30\u20132000 m. Voucher: Mwadime et al. 30 (EA).Sporobolusolivaceus Napper \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 2010\u20134000 m. Voucher: Turner 1568 (EA).Sporobolusquadratus Clayton \u2013 Life form: Herb. Habitat: Grassland and disturbed sites, 2200\u20133000 m. Voucher: Turner 1571 (EA).Sporobolusspicatus (Vahl) Kunth \u2013 Life form: Herb. Habitat: Grassland and open bushland, 0\u20132600 m. Voucher: Hammilton 384 (EA).Stipadregeana Steud. \u2013 Life form: Herb. Habitat: Moist evergreen forest, 1940\u20132770 m. Voucher: Robertson 7342 (EA).Streblochaetelongiarista (A.Rich) Pilg. \u2013 Life form: Herb. Habitat: Undergrowth in moist forest and bamboo thickets, 1500\u20133280 m. Voucher: Magogo 1529 (EA).Themedatriandra Forssk. \u2013 Life form: Herb. Habitat: Open deciduous bushland, 0\u20133200 m. Voucher: Turner 1577 (EA).Vulpiabromoides (L.) Gray \u2013 Life form: Herb. Habitat: Rocky grounds in upland grassland, 2500\u20133500 m. Voucher: Kuchar 12326 (EA).Yushaniaalpina (K.Schum.) W.C.Lin \u2013 Life form: Herb or subshrub. Habitat: Upland moist bush thickets and forested slopes, 2300\u20133300 m. Voucher: Fries 2565 (EA).PotamogetonaceaeF36. Potamogetonpusillus L. \u2013 Life form: Herb. Habitat: Aquatic in slow flowing streams, 600\u20132000 m. Voucher: Colgahoum (EA).Potamogetonrichardii Solms \u2013 Life form: Herb. Habitat: In water pools and streams, 1150\u20133450 m. Voucher: Verdcourt & Steele 913 (EA).SmilacaceaeF37. Smilaxaspera L. \u2013 Life form: Shrub. Habitat: Moist forest and associated bushland, 1450\u20132745 m. Voucher: Luke 14245 (EA).XanthorrhoeaceaeF38. Aloekedongensis Reynolds \u2013 Life form: Shrub. Habitat: Rocky sites in open woodland, 1825\u20132300 m. Voucher: Robertson 1747 (EA).Aloengongensis Christian \u2013 Life form: Shrub. Habitat: Open deciduous woodland and forest margins, 1370\u20131900 m. Voucher: Perdue and Kibuwa 8068 (EA).Aloenyeriensis Christian & I.Verd. \u2013 Life form: Shrub. Habitat: Upland open bushland, 1760\u20132100 m. Voucher: Napier 2186 (EA).Kniphofiathomsonii Baker \u2013 Life form: Herb. Habitat: Along stream and swampy sites in forest, 1850\u20133960 m. Voucher: SK 0060 .XyridaceaeF39. Xyriscapensis Thunb. \u2013 Life form: Herb. Habitat: Bogs and marshes in montane forest, 1100\u20133000 m. Voucher: Gilbert 4875 (EA).AcanthaceaeF40. Asystasialorata Ensermu \u2013 Life form: Herb. Habitat: Wooded grassland and bushland, 1400\u20132000 m. Voucher: Verdcourt 3553 (EA).Barleriaventricosa Hochst. ex Nees \u2013 Life form: Herb or subshrub. Habitat: Upland dry woodland and thickets, 1500\u20133590 m. Voucher: Napier 10542 (EA).Crossandratridentata Lindau \u2013 Life form: Herb. Habitat: Upland rainforest in shaded places, 700\u20132700 m. Voucher: Napier 2711 (EA).Diclipteralaxata C.B.Clarke \u2013 Life form: Herb or subshrub. Habitat: Moist montane forest and forest margins, 1300\u20132800 m. Voucher: Luke 9586 (EA).Diclipteramaculatasubsp.usambarica (Lindau) I.Darbysh. \u2013 Life form: Herb. Habitat: Moist forest, 1150\u20133050 m. Vouchers: Napier 620, Luke 8243 (EA).Dyschoristekeniensissubsp.keniensis Malombe, Mwachala & Vollesen \u2013 Life form: Herb or subshrub. Habitat: Wooded grassland and bushland, 900\u20132500 m. Voucher: Perdue and Kibuwa 8182 (EA).Dyschoristenagchana (Nees) Bennet \u2013 Life form: Herb. Habitat: Wet grasslands and swamps, 0\u20131800 m. Voucher: Faden 74/697 (EA).Dyschoristeradicans (Hochst. ex A.Rich) Nees \u2013 Life form: Herb or subshrub. Habitat: Upland bushland and grassland, 900\u20132500 m. Voucher: Mailnnes 69 (EA).Hypoestesaristata (Vahl) Roem. & Schult. \u2013 Life form: Herb or subshrub. Habitat: Forest margin, 200\u20133000 m. Voucher: Napier 619 (EA).Hypoestesforskaolii (Vahl) R.Br. \u2013 Life form: Herb or subshrub. Habitat: Dry grassland and bushland, 0\u20133000 m. Voucher: Nattrass 646 (EA).Hypoestestriflora (Forssk.) Roem. & Schult. \u2013 Life form: Herb. Habitat: Forest margins and grassland, 900\u20133200 m. Voucher: Albrechtren 2618 (EA).Isoglossagregoryi (S.Moore) Lindau \u2013 Life form: Herb. Habitat: Forest margins and grassland, 1700\u20132900 m. Voucher: Chandler 2221 (EA).Isoglossalactea Lindau ex Engl. \u2013 Life form: Herb or subshrub. Habitat: Montane forest, 1250\u20132350 m. Voucher: Taylor 1032 (EA).Isoglossasubstrobilina C.B.Clarke \u2013 Life form: Herb or subshrub. Habitat: Undergrowth in montane forest, 1750\u20132600 m. Voucher: Balbo 828 (EA).Justiciastriata (Klotzsch) Bullock \u2013 Life form: Herb. Habitat: Grassland and bushland, 200\u20132600 m. Voucher: Verdcourt 2651 (EA).Justiciaunyorensis S.Moore \u2013 Life form: Herb. Habitat: Moist woodland and bushland, 1150\u20133100 m. Voucher: Kerfoot 1389 (EA).Mimulopsisalpina Chiov. \u2013 Life form: Woody herb or shrub. Habitat: Montane evergreen forest, 1900\u20133300 m. Voucher: Dale 2154 (EA).Mimulopsissolmsii Schweinf. \u2013 Life form: Herb. Habitat: Montane evergreen forest, 1200\u20132700 m. Voucher: Jackson (EA).Phaulopsisimbricata (Forssk.) Sweet \u2013 Life form: Herb. Habitat: Forest edges and riverine forest, 1500\u20132970 m. Voucher: Faden 74/608 (EA).Rhinacanthusndorensis Schweinf. ex Engl. \u2013 Life form: Herb or subshrub. Habitat: Upland grassland and open woodland, 1700\u20132150 m. Voucher: Napier 2579 (EA).Thunbergiagibsonii S.Moore \u2013 Life form: Herbaceous climber. Habitat: Montane grassland and bushland, 1700\u20133000 m. Voucher: Fries 2815 (EA).Thunbergiaalata Bojer ex Sims \u2013 Life form: Herbaceous climber. Habitat: Wet bushland, 100\u20133000 m. Voucher: SK 0018 .Thunbergiafischeri Engl. \u2013 Life form: Herb. Habitat: Upland grassland, 1200\u20132500 m. Voucher: Napier 1798 (EA).Thunbergiareniformis Vollesen \u2013 Life form: Herbaceous climber. Habitat: Grassland, 1800\u20132400 m. Voucher: Someren 5057 (EA).AdoxaceaeF41. Sambucusafricana Standl. \u2013 Life form: Herb or shrub. Habitat: Roadsides in bamboo zone and montane forest, 1750\u20133370 m. Voucher: SK 0046 .Sambucusebulus L. \u2013 Life form: Herb. Habitat: Clearings and edges of moist bamboo thickets, 2320\u20133370 m. Voucher: Williams 1305 (EA).AmaranthaceaeF42. Achyranthesaspera L. \u2013 Life form: Herb. Habitat: Moist forest and open grassland, 0\u20133080 m. Voucher: SK 0206 .Aervalanata (L.) Juss. \u2013 Life form: Herb. Habitat: Moist forest edges and bushland, 0\u20132200 m. Voucher: Kirika et al. 152 (EA).Alternantheracaracasana Kunth \u2013 Life form: Herb. Habitat: Roadsides and forest edges, 0\u20132020 m. Voucher: Kokwaro 2795 (EA).Alternantherapungens Kunth \u2013 Life form: Herb. Habitat: Roadsides and river banks, 0\u20132020 m. Voucher: Bally 4609 (EA).Amaranthusgraecizans L. \u2013 Life form: Herb. Habitat: Grassland, 950\u20132900 m. Voucher: Hansen 85 (EA).Amaranthushybridus L. \u2013 Life form: Herb. Habitat: Roadsides and forest edges, 1500\u20132600 m. Voucher: Mbevi 182 (EA).Celosiaanthelminthica Asch. \u2013 Life form: Herb or subshrub. Habitat: Moist forest margins and clearings, 500\u20132300 m. Voucher: Someren 810 (EA).Chenopodiumalbum L. \u2013 Life form: Herb. Habitat: Roadsides in moist forest, 1650\u20132600 m. Voucher: Lochhead 610 (EA).Chenopodiumcarinatum R.Br. \u2013 Life form: Herb. Habitat: Roadsides and pasture grounds, 900\u20132100 m. Voucher: Verdcourt 391 (EA).Chenopodiumfasciculosum Aellen \u2013 Life form: Herb. Habitat: Roadsides in moist forest, 1310\u20132600 m. Voucher: Bogdan 4169 (EA).Chenopodiummurale L. \u2013 Life form: Herb. Habitat: Grazing areas in lower parts of montane forest, 1070\u20132750 m. Voucher: Mungai 57 (EA).Chenopodiumopulifolium Schrad. ex W.D.J.Koch & Ziz \u2013 Life form: Herb. Habitat: Roadsides and forest edges, 760\u20132100 m. Voucher: Bally 14439 (EA).Cyathulacylindrica Moq. \u2013 Life form: Herb or subshrub. Habitat: Moist forest and open rocky bushland, 1300\u20133240 m. Voucher: SK 0258 .Cyathulauncinulata (Schrad.) Schinz \u2013 Life form: Herb. Habitat: Grassland and open bushland, 900\u20132730 m. Voucher: Faden 74/606 (EA).Dysphaniaschraderiana (Schult.) Mosyakin & Clemants \u2013 Life form: Herb. Habitat: Roadsides and forest margins, 1600\u20132300 m. Voucher: Fries 138 (EA).Gomphrenacelosioides Mart. \u2013 Life form: Herb. Habitat: Forest edges and roadsides, 0\u20132150 m. Voucher: SK 0156 .Pupalialappacea (L.) Juss. \u2013 Life form: Herb. Habitat: Open sites in forest and forest edges, 10\u20132060 m. Voucher: Faden 74/664 (EA).AnacardiaceaeF43. Rhuslongipes Engl. \u2013 Life form: Shrub or small tree. Habitat: Evergreen bushland and forest margins, 1000\u20132400 m. Vouchers: Jackson 312, Gardner 3623 (EA).Sclerocaryabirrea (A.Rich) Hochst. \u2013 Life form: Tree. Habitat: Roadsides and bushland, 800\u20131800 m. Voucher: SK 0167 .Searsianatalensis (Bernh. ex C.Krauss) F.A.Barkley \u2013 Life form: Tree. Habitat: Evergreen bushland and forest edges, 1\u20133000 m. Voucher: SK 0099 .ApiaceaeF44. Afroligusticumaculeolatum (Engl.) P.J.D.Winter \u2013 Life form: Herb. Habitat: Bushland and moist montane forest, 1360\u20133030 m. Vouchers: Albrechtsen 6744, Scheffler 277 (EA).Afroligusticumelgonense (H.Wolff) P.J.D.Winter \u2013 Life form: Herb. Habitat: Wet sites and forest margins in montane forest, 1600\u20133600 m. Voucher: SK 0175 .Afroligusticumlinderi (C.Norman) P.J.D.Winter \u2013 Life form: Herb. Habitat: Montane grassland, 1860\u20133422 m. Voucher: Verdcourt & Polhill 7 (EA).EAfrosciadiumenglerianum (H.Wolff) P.J.D.Winter \u2013 Life form: Herb. Habitat: Tussocky grounds in moorland, 3400\u20133750 m. Voucher: Townsend 2406 (EA).EAfrosciadiumfriesiorumvar.friesiorum (H.Wolff) P.J.D.Winter \u2013 Life form: Herb. Habitat: Wet grassy sites in bamboo and ericaceous zone, 2950\u20134260 m. Voucher: SK 0112 .EAfrosciadiumfriesiorumvar.bipinnatum (C.C.Towns.) P.J.D.Winter \u2013 Life form: Herb. Habitat: Damp sites in moorland, 3000\u20133824 m. Voucher: Williams 1296 (EA).Afrosciadiumkerstenii (Engl.) P.J.D.Winter \u2013 Life form: Herb. Habitat: Open places in bamboo zone, 2550\u20134300 m. Voucher: Verdcourt 2055 (EA).Agrocharisincognita (C.Norman) Heywood & Jury \u2013 Life form: Herb. Habitat: Montane forest margins, 900\u20133600 m. Vouchers: SK 0223, SK 0180 .Agrocharismelanantha Hochst. \u2013 Life form: Herb. Habitat: Upland moist forest, 1520\u20133580 m. Voucher: RV 648 (EA).Alepideapeduncularis Steud. ex A.Rich. \u2013 Life form: Herb. Habitat: Grassland and open grounds in montane forest, 1050\u20133600 m. Voucher: SK 0109 .Anthriscussylvestris (L.) Hoffm. \u2013 Life form: Herb. Habitat: Forest margins and clearings in montane forest, 1800\u20133970 m. Voucher: Hedberg 1643 (EA).Berulaerecta (Huds.) Coville \u2013 Life form: Herb. Habitat: Stream banks and swampy sites, 1000\u20131900 m. Voucher: Fries & Fries 216 (EA).Centellaasiatica (L.) Urb. \u2013 Life form: Herb. Habitat: Wet grassland and open grounds in moist forest, 0\u20133540 m. Voucher: Kayombo et al. 5290 (EA).Haplosciadiumabyssinicum Hochst. \u2013 Life form: Herb. Habitat: Moist montane forest and moorland, 2150\u20134600 m. Voucher: SK 0084 .Heracleumabyssinicum (Boiss.) C.Norman \u2013 Life form: Herb. Habitat: Upland moist grassland, 1680\u20133970 m. Voucher: Townsend 2421 (EA).Heracleumelgonense (H.Wolff) Bullock \u2013 Life form: Herb. Habitat: Damp sites in moorland, 1080\u20134200 m. Voucher: SK 0115 .Heracleumtaylorii C.Norman \u2013 Life form: Herb. Habitat: Moorland, 3000\u20133800 m. Voucher: Taylor 1454 (EA).Oenanthepalustris (Chiov.) C.Norman \u2013 Life form: Herbaceous climber. Habitat: Moist forest edges, 1130\u20133260 m. Voucher: Carmichael 1293 (EA).Oenantheprocumbens (H.Wolff) Norman \u2013 Life form: Herbaceous climber. Habitat: Moist montane forest and shades in bamboo thickets, 1360\u20133200 m. Voucher: Townsend 2201 (EA).Oreoschimperellaaberdarensis (Norman) Rauschert \u2013 Life form: Herb. Habitat: Along streams in upland forest, 2200\u20132910 m. Voucher: Napier 605 (EA).Peucedanumelgonense H.Wolff \u2013 Life form: Herb. Habitat: Wet grounds in montane forest, 1600\u20133600 m. Vouchers: Greenway 10397 (EA), SK 0175 .Pimpinellakeniensis C.Norman \u2013 Life form: Herb. Habitat: Wooded grassland, 1550\u20132500 m. Voucher: SK 0058 .Pimpinellaoreophilavar.oreophila Hook.f. \u2013 Life form: Herb. Habitat: Moist montane forest, 2300\u20134100 m. Voucher: Townsend 2428 (EA).Pimpinellaoreophilavar.kilimandscharica (Engl.) C.C.Towns. \u2013 Life form: Herb. Habitat: Glades and grassland in upper montane forest, 3040\u20134030 m. Voucher: Townsend 2200 (EA).Pseudocarumeminii (Engl.) H.Wolff \u2013 Life form: Herbaceous climber. Habitat: Bamboo thickets, 1750\u20133350 m. Voucher: Polhill 241 (EA).Saniculaelata Buch.-Ham. ex D.Don. \u2013 Life form: Herb. Habitat: Shaded sites in bamboo thickets, 1240\u20133220 m. Voucher: Napier 608 (EA).Torilisafricana Spreng. \u2013 Life form: Exotic herb. Habitat: Riverine forest and forest edges, 1360\u20132720 m. Vouchers: Semsei 2811, Bytebier 264 (EA).*ApocynaceaeF45. Acokantheraschimperi (A.DC.) Benth. & Hook.f. ex Schweinf. \u2013 Life form: Shrub or small tree. Habitat: Dry forest margins, 250\u20132200 m. Voucher: Kamau 192 (EA).Asclepiasphysocarpa (E. Mey.) Schltr. \u2013 Life form: Exotic herb or subshrub. Habitat: Roadsides in montane forest, 1500\u20133000 m. Voucher: SK 0073 .* EBrachystelmakeniense Schweinf. \u2013 Life form: Herb. Habitat: Upland dry grassland, 1600\u20132700 m. Voucher: Hansen 79 (EA).Carissaspinarum L. \u2013 Life form: Shrub. Habitat: Bushland and riverine forest, 0\u20132250 m. Voucher: Kokwaro 2785 (EA).Cynanchumabyssinicum Decne. \u2013 Life form: Herbaceous climber. Habitat: Bushland and forest margins, 1600\u20132600 m. Vouchers: Polhill 433, Gilbert 6345 (EA).Cynanchumaltiscandens K.Schum. \u2013 Life form: Herbaceous climber. Habitat: Forest margins, 1200\u20132500 m. Voucher: Mathenge 599 (EA).Cynanchumviminalesubsp.suberosum (Meve & Liede) Goyder \u2013 Life form: Herbaceous climber. Habitat: Dry rocky grounds in forest, 100\u20132200 m. Voucher: Rayner 70 (EA).Dregeaschimperi (Decne.) Bullock \u2013 Life form: Woody climber. Habitat: Upland forest margins, 1500\u20132650 m. Voucher: SK 0001 .Gomphocarpuskaessneri (N.E.Br.) Goyder & Nicholas \u2013 Life form: Herb. Habitat: Seasonal wet pastures, 900\u20132300 m. Voucher: Faden 71/530 (EA).Gomphocarpussemilunatus A.Rich. \u2013 Life form: Herb. Habitat: Seasonal wet grassland, 1300\u20132650 m. Voucher: SAJIT 006505 .Gomphocarpusstenophyllus Oliv. \u2013 Life form: Herb. Habitat: Rocky sites in upland forest, 1200\u20133050 m. Voucher: Hooper et al 1647 (EA).Mondiawhitei (Hook.f.) Skeels \u2013 Life form: Woody climber. Habitat: Upland moist forest, 1600\u2013 2000 m. Voucher: Bell 1 (EA).Orbeasprengeri (Schweinf.) Bruyns \u2013 Life form: Herb. Habitat: Upland open woodland, 1400\u20132100 m. Voucher: Perkins s.n. (EA).Pachycarpusconcolor E.Mey. \u2013 Life form: Herb. Habitat: Upland swampy grassland, 1450\u20132100 m. Voucher: Faden et al. 74/574 (EA).Cynanchumethiopicumsubsp.angolense (N.E.Br.) Liede & Khanum \u2013 Life form: Herbaceous climber. Habitat: Moist forest, 1400\u20132200 m. Voucher: Napier 2475 (EA).Cynanchumgonoloboides Schltr. \u2013 Life form: Herbaceous climber. Habitat: Montane forest, 2400\u20133600 m. Voucher: SK 0196 .Pergulariadaemia (Forssk) Chiov. \u2013 Life form: Herbaceous climber. Habitat: Dry bushland, 0\u20132000 m. Voucher: SK 0207 .Periplocalinearifolia Quart.-Dill. & A.Rich. \u2013 Life form: Herbaceous climber. Habitat: Upland forest edges and riparian scrub forest, 1900\u20132900 m. Voucher: SK 0198 .Secamonealpini Schult. \u2013 Life form: Woody climber. Habitat: Montane forest, 1300\u20132150 m. Voucher: Luke et al. 8923 (EA).Secamonepunctulata Decne. \u2013 Life form: Woody climber. Habitat: Bush thickets and riverine forest, 0\u20132400 m. Voucher: Someren 15954 (EA).Tabernaemontanastapfiana Britten \u2013 Life form: Tree. Habitat: Moist forest, 1400\u20132370 m. Voucher: Beentje 2721 (EA).Tacazzeaconferta N.E.Br. \u2013 Life form: Woody climber. Habitat: Moist bushlands, 1500\u20133000 m. Voucher: Gillett 16649 (EA).Tylophoraanomala N.E.Br. \u2013 Life form: Herbaceous climber. Habitat: Margins of montane forest, 2000\u20132500 m. Voucher: SK 0251 .Tylophoraheterophylla A.Rich. \u2013 Life form: Woody climber. Habitat: Moist montane forest, 2200\u20133000 m. Voucher: Mbale et al. 858 (EA).Tylophoralugardae Bullock \u2013 Life form: Herbaceous climber. Habitat: Margins of montane forest, 2000\u20132500 m. Voucher: Kirrika 158 (EA).AquifoliaceaeF46. Ilexmitis (L.) Radlk. \u2013 Life form: Tree. Habitat: Upland moist forest, 900\u20133150 m. Voucher: Beentje 2721 (EA).AraliaceaeF47. Cussoniaholstii Harms ex Engl. \u2013 Life form: Tree. Habitat: Dry evergreen forest, 1110\u20132550 m. Voucher: SK 0270 .Hydrocotylemannii Hook.f. \u2013 Life form: Herb. Habitat: Roadsides and forest margins, 600\u20133370 m. Voucher: Luke 3887 (EA).Hydrocotyleranunculoides L.f. \u2013 Life form: Herb. Habitat: Along streams and swamps, 700\u20132400 m. Voucher: Kibui et al. 2867 (EA).Hydrocotylesibthorpioides Lam. \u2013 Life form: Herb. Habitat: Bogs, damp grassland and swamps in montane forest, 1134\u20133900 m. Voucher: SAJIT 006468 .Polysciasfulva (Hiern) Harms \u2013 Life form: Tree. Habitat: Upland moist forest and riverine forest, 1180\u20132300 m. Voucher: SK 0164 .Polysciaskikuyuensis Summerh. \u2013 Life form: Tree. Habitat: Upland moist forest, 1750\u20132620 m. Voucher: Kirika 28 (EA).Scheffleramyriantha (Baker) Drake \u2013 Life form: Woody climber. Habitat: Moist bamboo thickets, 1540\u20132770 m. Voucher: Verdcourt et al. 2974 (EA).Scheffleravolkensii (Harms) Harms \u2013 Life form: Tree. Habitat: Upland moist forest, 1550\u20133230 m. Voucher: Muchiri 588 (EA).AristolochiaceaeF48. Aristolochialittoralis Parodi \u2013 Life form: Woody climber. Habitat: Riverine forest, 1080\u20131800 m. Voucher: Gachathi 76/135 (EA).AsteraceaeF49. Acanthospermumglabratum (DC.) Wild \u2013 Life form: Exotic herb. Habitat: Roadsides in moist forest, 150\u20131850 m. Voucher: Mbevi 177 (EA).*Acmellacaulirhiza Delile \u2013 Life form: Herb. Habitat: Along streams and forest margins, 600\u20132600 m. Voucher: SK 0160 .Anthemistigrensis J. Gay ex A.Rich. \u2013 Life form: Herb. Habitat: Montane grassland and moorland, 1950\u20134300 m. Vouchers: SAJIT 006473, SK 0079 .Artemisiaafra Jacq. ex Willd \u2013 Life form: Woody herb. Habitat: Montane forest and moorland, 1500\u20134050 m. Voucher: Molony s.n. (EA).Baccharoideslasiopus (O.Hoffm.) H.Rob. \u2013 Life form: Shrub. Habitat: Roadsides and forest margins, 1100\u20132800 m. Voucher: Mungai 123(EA).Berkheyaspekeana Oliv. \u2013 Life form: Woody herb. Habitat: Rocky slopes in upland forest, 1800\u20133140 m. Voucher: Napier 1272 (EA).Bidenscinerea Sherff \u2013 Life form: Herb. Habitat: Dry bushland, 900\u20131950 m. Voucher: Verdcourt 3198 (EA).Bidenspilosa L. \u2013 Life form: Herb. Habitat: Forest margins and grassland, 750\u20132500 m. Voucher: Kerfoot 643 (EA).Bidensrueppellii (Sch.Bip. ex Sch.Bip.) Sherff \u2013 Life form: Herb or subshrub. Habitat: Moist forest margins, 2250\u20132800 m. Voucher: Pierce 2568 (EA).Bidenswhytei Sherff \u2013 Life form: Herb. Habitat: Riverine vegetation and moist forest margin, 750\u20132500 m. Voucher: Verdcourt 1479 (EA).Blumeaaxillaris (Lam.) DC. \u2013 Life form: Herb. Habitat: Swampy sites in grassland, 0\u20132400 m. Voucher: SK 0067 .Blumeaelatior (R.E.Fr.) Lisowski \u2013 Life form: Herb. Habitat: Swampy sites in grassland, 0\u20132400 m. Voucher: SK 0264 .Bothrioclineamplifolia (O.Hoffm. & Muschl.) M.G.Gilbert \u2013 Life form: Woody herb or shrub. Habitat: Montane forest, 2100\u20132850 m. Voucher: Mungai 89/171 (EA).Bothrioclinefusca (S.Moore) M.G.Gilbert \u2013 Life form: Shrub. Habitat: Montane forest, 1800\u20133600 m. Voucher: SAJIT 006475 .Bothrioclineglabrescens C.Jeffrey \u2013 Life form: Shrub. Habitat: Montane forest, 1950\u20133050 m. Voucher: Schmitt and Mathenge 1061 (EA).Bothrioclinelongipes (Oliv. & Hiern) N.E.Br. \u2013 Life form: Woody herb or shrub. Habitat: Upland grassland and forest margins, 850\u20132700 m. Voucher: Waiganjo 36 (EA).Carduusafromontanus R.E.Fr. \u2013 Life form: Herb. Habitat: Moist bamboo thicket, 2350\u20133430 m. Voucher: Agnew et al. 5623 (EA)Carduuskeniensis R.E.Fr. \u2013 Life form: Herb. Habitat: Moist sites in moorland, 2950\u20134570 m. Voucher: Leakey 1226 (EA).ECarduusmillefolius R.E.Fr. \u2013 Life form: Herb. Habitat: Moorland and open grounds in montane forest, 2750\u20133200 m. Voucher: Kuchar 9579 (EA).Carduusnyassanussubsp.nyassanus (S.Moore) R.E.Fr. \u2013 Life form: Herb. Habitat: Upland moist grassland, 1650\u20133150 m. Voucher: Kerfoot 650 (EA).Carduusnyassanussubsp.kikuyorum (R.E.Fr.) C.Jeffrey \u2013 Life form: Herb. Habitat: Moorland and upper margins of bamboo thickets, 1500\u20133450 m. Voucher: Kuchar 12343 (EA).Carduusschimperisubsp.nanus (R.E.Fr.) C.Jeffrey \u2013 Life form: Herb. Habitat: Grassland and moorland, 2550\u20134050 m. Vouchers: Coe 774, Meinertyhegen AH (EA).Carduussilvarum R.E.Fr. \u2013 Life form: Herb. Habitat: Upland dry forest, 1700\u20132300 m. Voucher: Verdcourt 3268 (EA).Centrapaluspauciflorus (Willd.) H.Rob. \u2013 Life form: Herb. Habitat: Dry forest margins, 800\u20132200 m. Voucher: Mungai 119 (EA).Cinerariadeltoidea Sond. \u2013 Life form: Herb. Habitat: Montane forest and forest edges, 1890\u20134050 m. Vouchers: SAJIT 006474, SK 0080, 0179 & 0267 .Cirsiumvulgare (Savi) Ten. \u2013 Life form: Herb. Habitat: Roadsides in upland grassland, 1790\u20132400 m. Voucher: Kuchar 9566 (EA).Conyzaclarenceana Oliv. & Hiern \u2013 Life form: Shrub. Habitat: Damp sites in moorland, 2300\u20133300 m. Voucher: Kuchar 10306 (EA).Conyzahochstetteri Sch.Bip. ex A.Rich. Life form: Herb. Habitat: Wooded grassland, 1050\u20133600 m. Voucher: Townsend (EA).Conyzahypoleuca A.Rich. \u2013 Life form: Woody herb or shrub. Habitat: Upland dry forest and forest margins, 1800\u20133000 m. Voucher: Kirika et al. 915 (EA).Conyzanewii Oliv. & Hiern \u2013 Life form: Woody herb or shrub. Habitat: Upland forest margins and bushlands, 1500\u20133050 m. Voucher: Luke 15354 (EA).Conyzapallidiflora R.E.Fr. \u2013 Life form: Woody herb. Habitat: Upland moist forest and river banks, 1800\u20133000 m. Voucher: SK 0181 .Conyzaruwenzoriensis (S.Moore) R.E.Fr. \u2013 Life form: Herb or shrub. Habitat: Afromontane grassland, 2500\u20133700 m. Voucher: Napier 599 (EA).Conyzaschimperi Sch.Bip. ex A.Rich. \u2013 Life form: Woody herb or shrub. Habitat: Upland grassland, 1650\u20133000 m. Voucher: Kirika et al. 1051 (EA).Conyzasubscaposa O.Hoffm. \u2013 Life form: Herb. Habitat: Afromontane grassland, 1200\u20134400 m. Voucher: Kerfoot 1519 (EA).Conyzatigrensis Oliv. & Hiern \u2013 Life form: Herb. Habitat: Roadsides in montane forest, 1350\u20133000 m. Voucher: Verdcourt & Lucas 320 (EA).Conyzavernonioides (Sch.Bip. ex A.Rich.) Wild \u2013 Life form: Shrub or small tree. Habitat: Montane forest margins, 2300\u20134000 m. Voucher: Battiscombe 946 (EA).Cotulaabyssinica Sch.Bip. ex A.Rich. \u2013 Life form: Herb. Habitat: Montane grassland and forest margins, 2100\u20133650 m. Voucher: Agnew and Timberlake 11121 (EA).Crassocephalumcrepidioides (Benth.) S.Moore \u2013 Life form: Herb. Habitat: Roadsides in moist forest, 0\u20132500 m. Voucher: Mutanga 33 (EA).Crassocephalummontuosum (S.Moore) Milne-Redh. \u2013 Life form: Herb or subshrub. Habitat: Upland moist forest, 900\u20133260 m. Voucher: Kerfoot 671 (EA).Crassocephalumvitellinum (Benth.) S.Moore \u2013 Life form: Herb. Habitat: Bushland and forest margins, 1500\u20132800 m. Voucher: Ericksson 607 (EA).Crepiscarbonaria Sch.Bip. \u2013 Life form: Herb. Habitat: Montane grassland, 2960\u20133850 m. Voucher: Kerfoot 1393 (EA).Crepisnewiisubsp.oliveriana (Kuntze) C.Jeffrey & Beentje \u2013 Life form: Herb. Habitat: Montane grassland, 1650\u20133750 m. Vouchers: Hedberg 1639, Coe & Kirika 422 (EA).Crepisrueppellii Sch.Bip. \u2013 Life form: Herb. Habitat: Upland grassland, 1300\u20133200 m. Vouchers: SAJIT 006470, SK 0173, SK 0056 .Dendroseneciobattiscombei (R.E.Fr. & T.C.E.Fr.) E.B.Knox \u2013 Life form: Shrub. Habitat: Wet sites in afro-alpine zones, 2950\u20134000 m. Voucher: Dale 6981 (EA).EDendroseneciobrassiciformis (R.E.Fr. & T.C.E.Fr.) Mabb. \u2013 Life form: Shrub. Habitat: Afro-alpine zone, 2950\u20133950 m. Voucher: Mabberley 379 (EA).EDendroseneciokeniensis (Baker f.) Mabb. \u2013 Life form: Shrub. Habitat: Afro-alpine zone, 3300\u20134275 m. Voucher: Faden 70/697 (EA).EDendroseneciokeniodendron (R.E.Fr. & T.C.E.Fr.) B.Nord. \u2013 Life form: Shrub. Habitat: Afro-alpine zone, 3650\u20134350 m. Voucher: Kirika et al. 51 (EA).Dichrocephalachrysanthemifoliavar.alpina (R.E.Fr.) Beentje \u2013 Life form: Herb. Habitat: Damp sites in moorland, 2200\u20133600 m. Vouchers: Bally 1227, Napier 625 (EA).Dichrocephalaintegrifolia (L.f.) Kuntze \u2013 Life form: Herb. Habitat: Moist forest margins and wooded grassland, 1100\u20133670 m. Voucher: Mwangangi 985 (EA).Echinopsaberdaricus R.E.Fr. \u2013 Life form: Herb. Habitat: Montane grassland and moorland, 2400\u20133450 m. Voucher: Kuchar 12706 (EA).Echinopsangustilobus S.Moore \u2013 Life form: Herb. Habitat: Bushed grassland, 2000\u20132850 m. Voucher: Kerfoot 2945 (EA).Echinopshoehnelii Schweinf. \u2013 Life form: Herb. Habitat: Open sites in montane forest, 2200\u20133500 m. Voucher: SK 0149 .Erigeronbonariensis L. \u2013 Life form: Exotic herb. Habitat: Roadsides in grassland, 300\u20132850 m. Voucher: Perdue and Kibuwa 8414 (EA).*Ethuliascheffleri S.Moore \u2013 Life form: Herb or subshrub. Habitat: Marshy grassland and river banks, 1500\u20132500 m. Voucher: Bally 13211 (EA).Euryopsbrownei S.Moore \u2013 Life form: Woody herb or shrub. Habitat: Upper parts of montane forest and heath zone, 2300\u20134000 m. Voucher: SK 0088 .Euryopschrysanthemoides (DC.) B.Nord. \u2013 Life form: Shrub. Habitat: Montane grassland and bushland, 1500\u20132100 m. Voucher: SK 0190 .Euryopsjacksonii S.Moore \u2013 Life form: Woody herb or shrub. Habitat: Montane grassland and bushland, 2000\u20133300 m. Voucher: Durie 1533 (EA).Gerberapiloselloides (L.) Cass. \u2013 Life form: Herb. Habitat: Wet woodland, moist forest and moorland, 900\u20133700 m. Voucher: Napier 646(EA).Gnaphaliumunionisvar.tweediae (Hilliard) Beentje \u2013 Life form: Herb. Habitat: Montane grassland, 1600\u20133200 m. Voucher: Townsend 2313 (EA).Guizotiajacksonii (S.Moore) J. Baag\u00f8e \u2013 Life form: Herb. Habitat: Montane forest glades and moorland, 2350\u20133900 m. Voucher: SK 0052 .Gutenbergiarueppellii Sch.Bip. \u2013 Life form: Herb. Habitat: Open woodland and grassland, 900\u20132300 m. Voucher: Rauh 500 (EA).Gymnanthemumurticifolium (A.Rich.) H.Rob. \u2013 Life form: Shrubland. Habitat: Upland forest margins, 2160\u20133000 m. Voucher: Greenway 9734 (EA).Gynurascandens O.Hoffm. \u2013 Life form: Herbaceous climber. Habitat: Moist forest margins and glades, 0\u20132200 m. Voucher: Napier 2452 (EA).Haplocarpharueppellii (Sch.Bip.) K.Lewin \u2013 Life form: Herb. Habitat: Moist montane grassland and moorland, 2550\u20134650 m. Voucher: SK 0083 .Helichrysumargyranthum O.Hoffm. \u2013 Life form: Woody herb or shrub. Habitat: Montane grassland and moorland, 2100\u20133900 m. Voucher: SAJIT 006477 .EHelichrysumbrownei S.Moore \u2013 Life form: Herb or subshrub. Habitat: Rocky slopes and ridges, 3300\u20134500 m. Voucher: SK 0086 (EA).Helichrysumchionoides Philipson \u2013 Life form: Shrub. Habitat: Moorland and bamboo zone, 2800\u20133850 m. Voucher: Coe & Kirika 327 (EA).Helichrysumcitrispinumvar.hoehnelii (Schweinf.) Schweinf. & Hedberg \u2013 Life form: Shrub. Habitat: Rocky sites in afro-alpine zone, 3200\u20135100 m. Vouchers: Young 5, Townsend 2402 (EA).Helichrysumellipticifolium Moeser \u2013 Life form: Herb or subshrub. Habitat: Damp sites in bamboo zone, 2500\u20134800 m. Voucher: Kuchar 12500 (EA).Helichrysumfoetidum (L.) Cass. \u2013 Life form: Herb. Habitat: Upland forest margins and clearings, 1350\u20133000 m. Voucher: Polhill et al. 319 (EA).Helichrysumformosissimum Sch.Bip. \u2013 Life form: Woody herb or shrub. Habitat: Swamps and bogs in moorland, 2300\u20134200 m. Voucher: Kuchar 12744 (EA).Helichrysumformosissimumvar.guilelmii (Engl.) Mesfin \u2013 Life form: Woody herb or shrub. Habitat: Wet sites in moorland, 1800\u20134200 m. Voucher: Kuchar 2694 (EA).Helichrysumforskahliivar.compactum (Vatke) Mesfin. \u2013 Life form: Woody herb or shrub. Habitat: Moorland, 1200\u20135000 m. Vouchers: Perdue and Kibuwa 8244, Kirika et al. 1063 (EA).Helichrysumglobosum Sch.Bip. \u2013 Life form: Herb. Habitat: Upland grassland and forest margins, 900\u20133100 m. Voucher: Coe 783 (EA).EHelichrysumgloria-dei Chiov. \u2013 Life form: Shrub. Habitat: Rocky sites in afro-alpine zone, 3650\u20134000 m. Voucher: Taylor 1495 (EA).Helichrysumkilimanjari Oliv. \u2013 Life form: Herb. Habitat: Montane grassland and heathland, 1650\u20133900 m. Voucher: Agnew 7205 (EA).Helichrysummaranguense O.Hoffm. \u2013 Life form: Shrub. Habitat: Upland forest margins and bamboo zone, 1950\u20132700 m. Voucher: Fries and Fries 2324 (EA).Helichrysumnewii Oliv. & Hiern \u2013 Life form: Shrub. Habitat: Montane grassland, 2700\u20134600 m. Voucher: Knox 2851 (EA).Helichrysumnudifolium (L.) Less. \u2013 Life form: Woody herb or shrub. Habitat: Upland grassland, 600\u20132750 m. Voucher: Faden & Evans 74/581 (EA).Helichrysumodoratissimum (L.) Sweet \u2013 Life form: Woody herb or shrub. Habitat: Montane grassland and bushland, 1700\u20133700 m. Voucher: Kuchar 12426 (EA).Helichrysumschimperi (Sch.Bip. ex A.Rich.) Moeser \u2013 Life form: Woody herb or shrub. Habitat: Upland grassland, 1350\u20133300 m. Voucher: Kokwaro and Mathenge 3271 (EA).Helichrysumsetosum Harv. \u2013 Life form: Herb. Habitat: Clearings in forest and grassland, 1250\u20133000 m. Voucher: Natrass 1375 (EA).Helichrysumstenopterum DC. \u2013 Life form: Herb. Habitat: Montane grassland and forest margins, 600\u20132800 m. Voucher: Kirika 161 (EA).Hilliardiellaaristata (DC.) H.Rob. \u2013 Life form: Herb. Habitat: Upland grassland and woodland, 1600\u20132400 m. Voucher: Faden 67/722 (EA).Hypochaerisglabra L. \u2013 Life form: Herb. Habitat: Roadsides in grassland, 1850\u20133000 m. Voucher: Kuchar 12290 (EA).Kleiniaabyssinicavar.hildebrandtii (Vatke) C.Jeffrey \u2013 Life form: Herb. Habitat: Dry grassland and woodland, 20\u20132700 m. Voucher: Bogdan 499 (EA).Lactucaglandulifera Hook.f. \u2013 Life form: Herb. Habitat: Moist forest and grassland, 1200\u20133600 m. Voucher: SK 0222 .Lactucainermis Forssk. \u2013 Life form: Herb. Habitat: Roadsides and disturbed sites in grassland, 500\u20133300 m. Voucher: Malombe & Kirika 22 (EA).Lactucaparadoxa Sch.Bip. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland forest margins and thickets, 1950\u20132650 m. Voucher: Kokwaro and Mathenge 3348 (EA).Laphangiumluteoalbum (L.) Tzvelev \u2013 Life form: Herb. Habitat: Montane grassland, 300\u20133850 m. Voucher: Muasya 2148 (EA).Linziaglabra Steetz \u2013 Life form: Woody herb. Habitat: Roadsides in bushland, 1300\u20132160 m. Voucher: Gilbert and Thulin 1750 (EA).Linziaituriensis (Muschl.) H.Rob. \u2013 Life form: Woody herb. Habitat: Moist forest, 1200\u20132500 m. Voucher: Verdcourt 3267 (EA).Melantherascandens (Schumach. & Thonn.) Roberty \u2013 Life form: Herb. Habitat: Riverine forest and swampy sites, 250\u20132200 m. Voucher: Perdue and Kibuwa 8201 (EA).Micractisbojeri DC. \u2013 Life form: Herb. Habitat: Moist forest and swamp grassland, 1350\u20132750 m. Voucher: Verdcourt et al. 3030 (EA).Microglossadensiflora Hook.f. \u2013 Life form: Shrub. Habitat: Montane forest margins, 1200\u20132650 m. Voucher: Mathenge 192 (EA).Microglossapyrifolia (Lam.) Kuntze \u2013 Life form: Shrub. Habitat: Grassland and bushland, 50\u20132650 m. Voucher: Kokwaro 327 (EA).Mikaniopsisbambuseti (R.E.Fr.) C.Jeffrey \u2013 Life form: Woody climber. Habitat: Bamboo zone, 2100\u20133150 m. Voucher: Vorontsova et al. 61 (EA).Mikaniopsisusambarensis (Muschl.) Milne-Redh. \u2013 Life form: Woody climber. Habitat: Moist forest margins, 850\u20132350 m. Voucher: Malombe et al. 27/986 (EA).Prenanthessubpeltata Stebbins \u2013 Life form: Herbaceous climber. Habitat: Bamboo thicket, 2470\u20132500 m. Voucher: Faden et al. 71/193 (EA).Pseudognaphaliumoligandrum (DC.) Hilliard & B.L.Burtt \u2013 Life form: Herb. Habitat: Montane grassland, 1050\u20132550 m. Voucher: Burney 18 (EA).Psiadiapunctulata (DC) Vatke \u2013 Life form: Woody herb or shrub. Habitat: Evergreen bushland and forest edges, 1000\u20132500 m. Voucher: SK 0010 .Schkuhriapinnata (Lam.) Kuntze ex Thell. \u2013 Life form: Exotic herb. Habitat: Upland dry woodland and grassland, 1000\u20132220 m. Voucher: Mbevi 189 (EA).*Senecioaequinoctialis R.E.Fr. \u2013 Life form: Woody herb. Habitat: Moist afro-alpine zone, 3000\u20134250 m. Voucher: SK 0085 .ESenecioamplificatus C.Jeffrey \u2013 Life form: Herb. Habitat: Upper regions of giant heath zone, 2900\u20133500 m. Voucher: Alexander 11635 (EA).Seneciocrispatipilosus C.Jeffrey \u2013 Life form: Herb. Habitat: Open sites in bamboo zone, 2050\u20133050 m. Voucher: Coe 754 (EA).Seneciodeltoideus Less. \u2013 Life form: Herb. Habitat: Moist forest margins, 1700\u20132600 m. Voucher: Kuchar 12245 (EA).Seneciohadiensis Forsk. \u2013 Life form: Herbaceous climber. Habitat: Riverine thickets and forest margins, 500\u20132600 m. Voucher: Hiepko 2678 (EA).Seneciohochstetteri Sch.Bip. ex A.Rich. \u2013 Life form: Herb. Habitat: Forest margins and wooded grassland, 900\u20133350 m. Voucher: Rauh 388 (EA).Seneciojacksonii S.Moore \u2013 Life form: Herb. Habitat: Montane swampy grassland, 3250\u20134150 m. Vouchers: Hedberg 1545, Rauh 2388 (EA).Seneciolyratus Forssk. \u2013 Life form: Woody climber. Habitat: Evergreen dry forest margins, 1500\u20132760 m. Voucher: Gillet 20736 (EA).Seneciomaranguensis O.Hoffm. \u2013 Life form: Woody herb or shrub. Habitat: Montane forest and bamboo thicket, 1800\u20133250 m. Voucher: Fries and Fries 2699 (EA).ESeneciomargaritae C.Jeffrey \u2013 Life form: Shrub. Habitat: Rocky bushland, 1800\u20131950 m. Voucher: Bally 1013 (EA).Seneciomesogrammoides O.Hoffm. \u2013 Life form: Herb. Habitat: Open grassland, 1200\u20132500 m. Voucher: SK 0044 .Seneciomoorei R.E.Fr. \u2013 Life form: Woody herb or shrub. Habitat: Montane grassland and heath zone, 1750\u20133500 m. Voucher: Kuchar 12527 (EA).Seneciopseudosubsessilis C.Jeffrey \u2013 Life form: Herb. Habitat: Upland forest margins and swamps, 1900\u20133050 m. Voucher: Charles 10580 (EA).ESenecioroseiflorus R.E.Fr. \u2013 Life form: Woody herb or shrub. Habitat: Rocky moorland and giant heath zone, 3100\u20134200 m. Vouchers: SAJIT 006490, SK 0114 .Senecioschweinfurthii O.Hoffm. \u2013 Life form: Herb. Habitat: Moorland and clearings in upper montane forest, 2300\u20134500 m. Voucher: SK 0089 .Seneciosubsessilis Oliv. & Hiern \u2013 Life form: Woody herb or shrub. Habitat: Montane forest margins, 1800\u20133600 m. Voucher: Kuchar 12550 (EA).Seneciosyringifolius O.Hoffm. \u2013 Life form: Herbaceous climber. Habitat: Montane forest margins, 1500\u20133300 m. Voucher: SK 0103 .Solanecioangulatus (Vahl) C.Jeffrey \u2013 Life form: Herbaceous climber. Habitat: Riverine vegetation and moist bushland, 1800\u20132500 m. Voucher: Faden et al. 74/657 (EA).Solaneciomannii (Hook.f.) C.Jeffrey \u2013 Life form: Shrub or small tree. Habitat: Forest glades and margins, 80\u20132700 m. Voucher: SK 0165 .Solanecionandensis (S.Moore) C.Jeffrey \u2013 Life form: Herbaceous climber. Habitat: Forest margins and bushland, 1500\u20132700 m. Voucher: Kamau 372 (EA).Sonchusafromontanus R.E.Fr. \u2013 Life form: Herb. Habitat: Upland grassland, 2200\u20133700 m. Voucher: Agnew et al. 142 (EA).Sonchusasper (L.) Hill \u2013 Life form: Herb. Habitat: Upland grassland, 1600\u20133000 m. Voucher: Someren 63/192 (EA).Sonchusbipontini Asch. \u2013 Life form: Herb. Habitat: Upland grassland, forest glades and margins, 1700\u20133370 m. Voucher: Hooper and Townsend 1389 (EA).Sonchuscamporum (R.E.Fr.) Boulos ex C.Jeffrey \u2013 Life form: Herb. Habitat: Upland grassland, 1800\u20132300 m. Voucher: Fries & Fries 513b (EA).Sonchusluxurians (R.E.Fr.) C.Jeffrey \u2013 Life form: Herb. Habitat: Roadsides in grassland and montane forest margins, 1600\u20133800 m. Voucher: Young 1001 (EA).Sonchusstenophyllus R.E.Fr. \u2013 Life form: Herb. Habitat: Upland grassland and forest edges, 2200\u20133700 m. Voucher: Fries & Fries 123 (EA).Sphaeranthussuaveolens (Forsk.) DC. \u2013 Life form: Herb. Habitat: Stream banks and marshy sites, 350\u20132500 m. Voucher: Kuchar 8276 (EA).Stoebekilimandscharica O.Hoffm. \u2013 Life form: Shrub. Habitat: Moist afro-alpine zone, 1950\u20133900 m. Voucher: Bally 1167 (EA).Tagetesminuta L. \u2013 Life form: Exotic herb. Habitat: Roadsides and disturbed sites, 850\u20132750 m. Voucher: Spjut 2791 (EA).*Taraxacumcampylodes G.E.Haglund \u2013 Life form: Herb. Habitat: Roadsides in montane forest, 2350\u20132500 m. Voucher: Dyson 638 (EA).Tithoniadiversifolia (Hemsl.) A.Gray \u2013 Life form: Exotic woody herb or subshrub. Habitat: Roadsides and forest margins, 0\u20131950 m. Voucher: SK 0204 .*Tolpiscapensis (L.) Sch.Bip. \u2013 Life form: Herb. Habitat: Open woodland and grassland, 1800\u20133300 m. Voucher: Kerfoot 1457 (EA).Tripterisvaillantii Decne. \u2013 Life form: Herb or subshrub. Habitat: Upland grassland and bushland, 1200\u20133000 m. Voucher: Faden et al. 74/646 (EA).Vernoniaauriculifera Hiern \u2013 Life form: Woody herb or shrub. Habitat: Montane forest margins and glades, 750\u20133000 m. Voucher: Kuchar 8342 (EA).Vernoniahochstetteri Sch.Bip. ex Walp. \u2013 Life form: Woody herb or shrub. Habitat: Moist forest and bushlands, 1000\u20132400 m. Voucher: Polhill 76 (EA).Vernoniapteropoda Oliv. & Hiern \u2013 Life form: Herb or subshrub. Habitat: Wet evergreen forest, 1800\u20132750 m. Voucher: Verdcourt 2311 (EA).Vernoniasubscandens R.E.Fr. \u2013 Life form: Shrub. Habitat: Moist forest and forest margins, 1650\u20132100 m. Voucher: Verdcourt 3706 (EA).Vernoniasyringifolia O.Hoffm. \u2013 Life form: Woody herb or shrub. Habitat: Moist forest and forest margins, 1550\u20133050 m. Voucher: Kerfoot 465 (EA).Vernoniaamygdalina Delile \u2013 Life form: Shrub or small tree. Habitat: Grassland and woodland, 200\u20132300 m. Voucher: SK 0016 .Vernoniagalamensissubsp.afromontana (R.E.Fr.) M.G.Gilbert \u2013 Life form: Shrub. Habitat: Dry forest margins and glades, 800\u20132200 m. Voucher: SK 0205 .BalsaminaceaeF50. Impatiensfischeri Warb. \u2013 Life form: Herb. Habitat: Moist shaded sites in upland forest, 2000\u20133100 m. Voucher: SK 0127 .Impatienshoehnelii T.C.E.Fr. \u2013 Life form: Herb. Habitat: Moist shaded sites in rainforest, 1475\u20133350 m. Voucher: SK 0126 .Impatiensmeruensissubsp.cruciata (T.C.E.Fr.) Grey-Wilson \u2013 Life form: Herb. Habitat: Moist forest and bamboo thickets, 1100\u20133630 m. Voucher: Napier 722 (EA).Impatienstinctoria A.Rich. \u2013 Life form: Herb. Habitat: Waterfalls and stream banks in wet upland forests, 1800\u20133630 m. Voucher: SK 0178 .BasellaceaeF51. Basellaalba L. \u2013 Life form Herbaceous climber. Habitat: Bush thickets and forest edges, 0\u20132450 m. Voucher: Faden 74/712 (EA).BegoniaceaeF52. Begoniameyeri-johannis Engl. \u2013 Life form: Woody climber. Habitat: Moist forest, 1350\u20132800 m. Vouchers: SK 0183, SK 0187 .BerberidaceaeF53. Berberisholstii Engl. \u2013 Life form: Shrub. Habitat: Upland forest margins and bushland 1500\u20133450 m. Voucher: Battiscombe 203 (EA).BignoniaceaeF54. Jacarandamimosifolia D.Don \u2013 Life form: Exotic tree. Habitat: Cultivated, 1970\u20131970 m. Voucher: Perdue and Kibuwa 8096 (EA).*Kigeliaafricanasubsp.moosa (Sprague) Bidgood & Verdc. \u2013 Life form: Tree. Habitat: Moist evergreen forest and swampy forest, 1050\u20132250 m. Vouchers: Perdue and Kibuwa 8421, Nyakundi 421 (EA).Spathodeacampanulata P.Beauv. \u2013 Life form: Tree. Habitat: Montane forest and riverine forest, 1500\u20132000 m. Voucher: SK 0202 .BoraginaceaeF55. Cordiaafricana Lam. \u2013 Life form: Tree. Habitat: Forest edges and wooded grassland, 450\u20132100 m. Voucher: SK 0224 .Cynoglossumaequinoctiale T.C.E.Fr. \u2013 Life form: Herb. Habitat: Upland grassland, 2100\u20132870 m. Voucher: Lacey 24A (EA).Cynoglossumamplifoliumvar.amplifolium Hochst. ex A.DC. \u2013 Life form: Herb. Habitat: Grassland and open sites in bamboo thickets, 1800\u20133200 m. Voucher: Brown 1717 (EA).Cynoglossumamplifoliumvar.subalpinum (T.C.E.Fr.) Verdc. \u2013 Life form: Herb. Habitat: Montane grassland and open sites in bamboo thickets, 2100\u20133430 m. Voucher: Kerfoot 1418 (EA).Cynoglossumcoeruleumvar.kenyense B. Verdcourt \u2013 Life form: Herb. Habitat: Grassland and forest edges, 1100\u20133200 m. Vouchers: SAJIT 006460, SK 0075, SK 0174 .Cynoglossumlanceolatum Forssk. \u2013 Life form: Herb. Habitat: Grassland and bushland, 1100\u20133220 m. Voucher: Kuchar 12236 (EA).Ehretiacymosavar.silvatica (G\u00fcrke) Brenan \u2013 Life form: Tree. Habitat: Moist forest and bushland, 960\u20132250 m. Vouchers: Kenya Forest excursion 88 (EA), SK 0154 .Heliotropiumscotteae Rendle \u2013 Life form: Herb. Habitat: Open sites in upland forests and bushland, 1500\u20132170 m. Voucher: McDonald 879 (EA).Heliotropiumzeylanicum (Burm.f.) Lam. \u2013 Life form: Herb. Habitat: Roadsides in grassland and bushland, 0\u20132317 m. Voucher: SK 0171 .Lithospermumafromontanum Weim. \u2013 Life form: Woody climber. Habitat: Montane forest margins, 1560\u20133950 m. Voucher: SAJIT 006483 .Myosotisabyssinica Boiss. & Reut. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 1560\u20133590 m. Voucher: Mabberley 381 (EA).Myosotisvestergrenii Stroh \u2013 Life form: Herb. Habitat: Wet sites in moorland and bamboo thickets, 2000\u20134250 m. Voucher: Rauh et al. 527 (EA).Trichodesmaambacensesubsp.hockii (De Wild.) Brummitt. \u2013 Life form: Herb. Habitat: Burnt grassland and woodland, 780\u20133000 m. Vouchers: Shayse 8835, Bally 8835 (EA).Trichodesmaphysaloides (Fenzl) A.DC. \u2013 Life form: Herb. Habitat: Wooded grassland, 700\u20132400 m. Voucher: SK 0184 (EA).BrassicaceaeF56. Arabidopsisthaliana (L.) Heynh. \u2013 Life form: Herb. Habitat: Montane bushland and moorland, 1750\u20134250 m. Voucher: Coe & Kirika 290 (EA).Arabisalpina L. \u2013 Life form: Herb. Habitat: Damp sites in moorland, 2450\u20134800 m. Voucher: SAJIT 006461 .Barbareaintermedia Boreau \u2013 Life form: Herb. Habitat: Streamsides in upper parts of montane forest, 3050\u20133950 m. Voucher: Muninentyhegen 9313 (EA).Brassicanapus L. \u2013 Life form: Exotic herb. Habitat: Escaped cultivation common along roadsides and disturbed sites, 1750\u20132300 m. Voucher: Someren s.n (EA).*Brassicarapa L. \u2013 Life form: Exotic herb. Habitat: Escape cultivation common along roadsides, 1500\u20132600 m. Voucher: Someren 603 (EA).*Capsellabursa-pastoris (L.) Medik. \u2013 Life form: Herb. Habitat: Roadsides in montane forest, 1600\u20132500 m. Voucher: Greenway 10206 (EA).Cardamineafricana L. \u2013 Life form: Herb. Habitat: Moist forest, 1000\u20133400 m. Voucher: Muninentyhegen 9304 (EA).Cardamineobliqua Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Moist montane forest up to moorland, 2000\u20134900 m. Voucher: Hedberg 1596 (EA).Cardaminehirsuta L. \u2013 Life form: Herb. Habitat: Wet open grounds in montane forest, 500\u20134600 m. Vouchers: SAJIT 006466, SK 0177 .Erucastrumarabicum Fisch. & C.A.Mey. \u2013 Life form: Herb. Habitat: Roadsides in upland forests, 0\u20133170 m. Voucher: John Terry 180 (EA).Farsetiastenoptera Hochst. \u2013 Life form: Herb. Habitat: Roadsides in open bushland, 500\u20133613 m. Voucher: SK 0094 .Farsetiaundulicarpa Jonsell \u2013 Life form: Shrub. Habitat: Bushland and wooded grassland, 1750\u20132300 m. Voucher: Verdcourt 2159 (EA).Lepidiumafricanum (Burm.f.) DC. \u2013 Life form: Herb. Habitat: Along the streams and roadsides in upland grassland, 1200\u20132120 m. Voucher: Harper 2143 (EA).Lepidiumbonariense L. \u2013 Life form: Exotic herb. Habitat: Roadsides in upland grassland, 1460\u20132650 m. Voucher: Verdcourt 2767 (EA).*Lepidiumdidymum L. \u2013 Life form: Herb. Habitat: Roadsides and forest clearings, 1350\u20132800 m. Voucher: Kroo 13173 (EA).Nasturtiummicrophyllum (Boenn. ex Rchb.) Rchb. \u2013 Life form: Herb. Habitat: Upland stream banks, 1500\u20132000 m. Voucher: Bally 8653 (EA).Nasturtiumofficinale R.Br. \u2013 Life form: Herb. Habitat: Muddy soils along rivers, 1500\u20132700 m. Voucher: Bogdan 2140 (EA).Oreophytonfalcatum O.E.Schulz \u2013 Life form: Herb. Habitat: Rocky sites in afro-alpine zone, 3820\u20134900 m. Voucher: Hedberg 1557 (EA).Raphanusraphanistrum L. \u2013 Life form: Exotic herb. Habitat: Roadsides and other ruderal sites, 15\u20132750 m. Voucher: Someren 549-556 (EA).*Raphanussativus L. \u2013 Life form: Exotic herb. Habitat: Roadsides, 15\u20132650 m. Voucher: Someren 550 (EA).*Rorippacryptantha (A.Rich.) Robyns & Boutique \u2013 Life form: Herb. Habitat: Stream banks and wet sites in forest, 1800\u20133000 m. Voucher: Bogdan 4757 (EA).Rorippamicrantha (Roth) Jonsell \u2013 Life form: Herb. Habitat: Along streams and muddy sites, 10\u20132400 m. Voucher: Gillett (EA).Rorippanudiuscula Thell. \u2013 Life form: Herb. Habitat: Upland moist forest and stream banks, 2200\u20133000 m. Voucher: Kuchar 8288 (EA).Sisymbriumerysimoides Desf. \u2013 Life form: Herb. Habitat: Clearings in forest and disturbed sites, 2000\u20132400 m. Voucher: Meinertzhagen 9309 (EA).Sisymbriumofficinale (L.) Scop. \u2013 Life form: Herb. Habitat: Roadsides and waste places, 1350\u20131800 m. Voucher: Frank Msafiri 20 (EA).Sisymbriumorientale L. \u2013 Life form: Herb. Habitat: Roadsides and waste places, 15\u20132000 m. Voucher: Greenway 14910 (EA).Subulariamonticola A.Braun ex Schweinf. \u2013 Life form: Herb. Habitat: Moist montane forest and moorland, 2750\u20134750 m. Voucher: SAJIT 006467 .Thlaspialliaceum L. \u2013 Life form: Herb. Habitat: Upper montane forest and moorland, 3050\u20133600 m. Voucher: Gillet 16229 (EA).Turritisglabra L. \u2013 Life form: Herb. Habitat: Roadsides in upland moist forest, 920\u20132700 m. Voucher: Verdcourt 3206 (EA).BurseraceaeF57. Commiphoraafricanavar.oblongifoliolata (Engl.) J.B.Gillett \u2013 Life form: Shrub or small tree. Habitat: Bushed grassland, 20\u20131890 m. Voucher: Adamson (EA).Commiphoraafricanavar.rubriflora (Engl.) Wild \u2013 Life form: Shrub or small tree. Habitat: Bushed grassland, 640\u20132070 m. Voucher: Gillett 19442 (EA).CampanulaceaeF58. Campanulaedulis Forssk. \u2013 Life form: Herb. Habitat: Upland grassland, 1500\u20133700 m. Voucher: Napier 2107 (EA).Canarinaeminii Asch. & Schweinf. \u2013 Life form: Herb. Habitat: Often epiphytic in moist forests, 1600\u20133200 m. Voucher: SK 0141 .Lobeliaaberdarica R.E.Fr. & T.C.E.Fr. \u2013 Life form: Shrub. Habitat: Swampy sites in montane forest, 1800\u20133500 m. Voucher: Rauh 520 (EA).Lobeliabambuseti R.E.Fr. & T.C.E.Fr. \u2013 Life form: Shrub. Habitat: Upland forests and bamboo thickets, 2350\u20134000 m. Voucher: Taylor 1361 (EA).Lobeliabaumannii Engl. \u2013 Life form: Shrub. Habitat: Forest floors and margins, 800\u20132400 m. Voucher: Knox 2575 (EA).Lobeliaduriprati T.C.E.Fr. \u2013 Life form: Herb. Habitat: Roadsides and forest margins, 1675\u20133550 m. Voucher: Agnew and Timberlake 11144 (EA).ELobeliagregorianasubsp.sattimae (R.E.Fr. & T.C.E.Fr.) E.B.Knox \u2013 Life form: Shrub. Habitat: Wet places in moorland and stream banks, 3350\u20133900 m. Vouchers: Hedberg 1608, Kirika and York 1192 (EA).Lobeliaholstii Engl. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 900\u20133520 m. Voucher: Kerfoot 412 (EA).Lobelialindblomii Mildbr. \u2013 Life form: Herb. Habitat: Moorland and swampy grounds in montane grassland, 3100\u20134250 m. Voucher: Knox 2519 (EA).Lobeliaminutula Engl. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 2125\u20133940 m. Voucher: Polhill 166 (EA).Lobeliatelekii Schweinf. \u2013 Life form: Shrub. Habitat: Upland grassland and moorland, 2950\u20134550 m. Voucher: Kirika et al. 905 (EA).Monopsisstellarioidessubsp.schimperiana (Urb.) Thulin \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 1100\u20133600 m. Vouchers: Kuchar 12413, Napier 730 (EA).Wahlenbergiacapillaceasubsp.tenuior (Engl.) Thulin \u2013 Life form: Herb. Habitat: Upland grassland and rocky sites in bamboo zone, 1500\u20133500 m. Vouchers: Gardner 10120, Leakey 14 (EA).Wahlenbergiakrebsiisubsp.arguta (Hook.f.) Thulin \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 1500\u20134000 m. Vouchers: Kuchar 9612, 10395 (EA).Wahlenbergiapusilla Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 2800\u20134500 m. Voucher: Hedberg 4297 (EA).Wahlenbergiascottii Thulin \u2013 Life form: Herb. Habitat: Upland grassland, 1500\u20133000 m. Voucher: Bogdan 4451 (EA).Wahlenbergiasilenoides Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland grassland and forest margins, 2200\u20133350 m. Voucher: Chandler 2317 (EA).Wahlenbergiavirgata Engl. \u2013 Life form: Herb. Habitat: Upland grassland, 1100\u20132700 m. Voucher: H & F 4850 (EA).CanellaceaeF59. Warburgiaugandensis Sprague \u2013 Life form: Tree. Habitat: Moist forest, 1100\u20132230 m. Voucher: Trapnell 2143 (EA).CannabaceaeF60. Celtisafricana Burm.f. \u2013 Life form: Tree. Habitat: Upland rainforest and riverine forest, 30\u20132400 m. Voucher: Kamau 418 (EA).CapparaceaeF61. Cadabafarinosa Forssk. \u2013 Life form: Shrub. Habitat: Deciduous bushland and grassland, 0\u20131900 m. Voucher: SK 0007 .Capparisfascicularisvar.elaeagnoides (Gilg) DeWolf \u2013 Life form: Woody climber. Habitat: Deciduous bushland and grassland, 900\u20132100 m. Vouchers: Battiscombe 1091 (EA), SK 0004 .Capparistomentosa Lam. \u2013 Life form: Shrub or small tree. Habitat: Bushland and grassland, 0\u20132500 m. Voucher: Someren 1769 (EA).Capparisviminea Oliv. \u2013 Life form: Shrub. Habitat: Moist forest, 0\u20132030 m. Voucher: Battiscombe 562 (EA).Maeruatriphylla A.Rich. \u2013 Life form: Shrub. Habitat: Grassland and bushland, 0\u20132300 m. Voucher: Agnew et al. 8724 (EA).Ritchieaalbersii Gilg \u2013 Life form: Tree. Habitat: Upland rainforest, 1100\u20132400 m. Voucher: SK 0155 .Thylaciumafricanum Lour. \u2013 Life form: Shrub or small tree. Habitat: Wooded grassland and deciduous bushland, 1\u20132400 m. Voucher: SK 0026 .CaprifoliaceaeF62. Dipsacuspinnatifidus Steud. ex A.Rich. \u2013 Life form: Herb. Habitat: Clearings in upland forests and bamboo thickets, 2000\u20133950 m. Voucher: Napier 664 .Scabiosacolumbaria L. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 2100\u20134100 m. Voucher: SK 0092 .Valerianacapensis Thunb. \u2013 Life form: Herb. Habitat: Upland moist forests and moorland, 1500\u20133400 m. Voucher: Coe 757 (EA).Valerianakilimandscharica Engl. \u2013 Life form: Woody herb. Habitat: Wet sites in moorland and tussocky grassland, 2800\u20134500 m. Voucher: Dale 294 (EA).Valerianellamicrocarpa Loisel. \u2013 Life form: Herb. Habitat: Moorlands and upper parts of bamboo zone, 2800\u20133500 m. Voucher: Kokwaro et al. 2423 (EA).CaryophyllaceaeF63. Cerastiumafromontanum T.C.E.Fr. \u2013 Life form: Herb. Habitat: Moorland, 2100\u20133940 m. Voucher: SK 0061 (EA).Cerastiumlanceolatum (Poir.) Volponi \u2013 Life form: Herb. Habitat: Forest margins and glades, 1050\u20133600 m. Voucher: Drummond and Hemsley 4283 (EA).Cerastiumoctandrumvar.adnivale (Chiov.) M\u00f6schl \u2013 Life form: Herb. Habitat: Montane bushland edges and open grassland, 1920\u20134200 m. Vouchers: Gillett 18988, Kuchar 12460 (EA).Corrigiolalitoralis L. \u2013 Life form: Herb. Habitat: Roadsides in montane forest, 1200\u20132190 m. Voucher: Mathenge 208 (EA).Drymariacordata (L.) Willd. ex Schult. \u2013 Life form: Herb. Habitat: Roadsides in forest and bushland, 870\u20132700 m. Voucher: SK 0136 .Saginaabyssinica Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Damp sites in moorland, 2150\u20134250 m. Voucher: Coe 784 (EA).Saginaafroalpina Hedberg \u2013 Life form: Herb. Habitat: Bogs and swamps in montane forest, 2980\u20134600 m. Voucher: Hedberg 1542 (EA).Sileneburchellii Otth ex DC. \u2013 Life form: Herb. Habitat: Rocky grounds in moorland, 1500\u20134050 m. Voucher: Kuchar 10349 (EA).Silenemacrosolen Steud. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland rocky grassland, 1800\u20133300 m. Voucher: Bally 906 (EA).Stellariasennii Chiov. \u2013 Life form: Herb. Habitat: Roadsides in upland wet forests, 1650\u20133440 m. Voucher: Young 1002 (EA).Uebeliniacrassifolia T.C.E.Fr. \u2013 Life form: Herb. Habitat: Grassy glades in bamboo thickets and moorland, 2500\u20134000 m. Voucher: Miss Dent 1306 (EA).CelastraceaeF64. Cassinebuchananii Loes. \u2013 Life form: Tree. Habitat: Dry evergreen forest and wooded grassland, 1000\u20132330 m. Voucher: Verdcourt 3049 (EA).Gymnosporiabuchananii Loes. \u2013 Life form: Shrub. Habitat: Dry evergreen forest, 60\u20132640 m. Voucher: Bogdan 467 (EA).Gymnosporiaheterophylla (Eckl. & Zeyh.) Loes. \u2013 Life form: Shrub. Habitat: Moist forest and riverine forest, 0\u20132670 m. Voucher: Ward 3049 (EA).Gymnosporiaputterlickioides Loes. \u2013 Life form: Shrub. Habitat: Dry woodland, 850\u20131800 m. Vouchers: H & J 6603 (EA) SK 0100 .Hippocrateagoetzei Loes. \u2013 Life form: Woody climber. Habitat: Evergreen forest, 0\u20133000 m. Voucher: SK 0247 .Maytenusobscura (A.Rich.) Cufod. \u2013 Life form: Shrub or small tree. Habitat: Moist forest, 2100\u20132550 m. Voucher: Bogdan 468 (EA).Maytenusundata (Thunb.) Blakelock \u2013 Life form: Tree. Habitat: Moist forest, 0\u20133150 m. Voucher: Kuchar 10284 (EA).Cassineaethiopicum (Thunb.) Loes. \u2013 Life form: Tree. Habitat: Moist forest, 0\u20132550 m. Voucher: Hansen 804 (EA).Pristimeragoetzei (Loes.) R. H. Archer \u2013 Life form: Herb or subshrub. Habitat: Moist forest, 90\u20133000 m. Voucher: Beentje and Mungai 2898 (EA).CleomaceaeF65. Cleomegynandra L. \u2013 Life form: Herb. Habitat: Roadsides and disturbed sites, 0\u20132400 m. Voucher: Ward 10861 (EA).ClusiaceaeF66. Garciniavolkensii Engl. \u2013 Life form: Tree. Habitat: Moist or dry evergreen forest, 30\u20132400 m. Voucher: Kuchar et al. 5455 (EA).ConnaraceaeF67. Agelaeapentagyna (Lam.) Baill. \u2013 Life form: Woody climber. Habitat: Upland wet forest, 1200\u20132100 m. Voucher: Ndonge 37 (EA).Roureathomsonii (Baker.) Jongkind \u2013 Life form: Shrub or small tree. Habitat: Upland wet forest, 0\u20132500 m. Voucher: Kuchar 5460 (EA).ConvolvulaceaeF68. Convolvulusfarinosus L -. Life form: Herbaceous climber. Habitat: Upland grassland, 450\u20132600 m. Voucher: Faden 67/270 (EA).Convolvuluskilimandschari Engl. \u2013 Life form: Herbaceous climber. Habitat: Moist forest margins and bamboo thicket, 1800\u20133750 m. Voucher: Svarreush 18 (EA).Convolvulussiculus L. \u2013 Life form: Herb. Habitat: Grassland, 1800\u20132300 m. Voucher: Faden 67/270 (EA).Cuscutaaustralis R.Br. \u2013 Life form: Herbaceous climber. Habitat: Parasitic in swampy vegetation, 1750\u20132170 m. Voucher: Gillett 16568 (EA).Cuscutakilimanjari Oliv. \u2013 Life form: Herbaceous climber. Habitat: Parasitic in upland moist forest, 500\u20132770 m. Voucher: Faden 74/709 (EA).Cuscutaplanifloravar.madagascarensis (Yunck.) Verdc. \u2013 Life form: Herbaceous climber. Habitat: Parasitic in upland grassland, 1500\u20133000 m. Vouchers: Bogdan 839, Gillett 16568 (EA).Dichondrarepens J.R.Forst. & G.Forst. \u2013 Life form: Herb. Habitat: Upland grassland, 1650\u20132520 m. Voucher: Faden 67418 (EA).Ipomoeaalba L. \u2013 Life form: Herbaceous climber. Habitat: Moist forest, 420\u20133393 m. Voucher: SAJIT 006487 .Ipomoeapurpurea (L.) Roth \u2013 Life form: Herbaceous climber. Habitat: Escaped cultivation common on roadsides and other waste places, 900\u20132040 m. Voucher: Greenway 10950 (EA).Ipomoeatenuirostrissubsp.tenuirostris Steud. ex Choisy \u2013 Life form: Herb. Habitat: Upland bushland, 1350\u20132250 m. Vouchers: Faden 68/283 (EA), SK 0048 .Ipomoeawightii Choisy \u2013 Life form: Herb. Habitat: Upland grassland and montane forest margins, 1040\u20132400 m. Voucher: Beentje 3195 (EA).CornaceaeF69. Cornusvolkensii Harms \u2013 Life form: Tree. Habitat: Moist forest and riparian forest, 1200\u20133000 m. Vouchers: Dale 402 (EA), SK 0131 .CrassulaceaeF70. Crassulaalatasubsp.pharnaceoides (Fisch. & C.A.Mey.) Wickens & Bywater \u2013 Life form: Herb. Habitat: Moist rocky sites and stream banks, 1900\u20132100 m. Vouchers: Hedberg 6278, Gilbert 4835 (EA).Crassulaalsinoides (Hook.f.) Engl. \u2013 Life form: Herb. Habitat: Along streams and swamps, 1300\u20133500 m. Voucher: Agnew 7164 (EA).Crassulagranvikii Mildbr. \u2013 Life form: Herb. Habitat: Along streams in alpine zone and damp open soils, 1200\u20134250 m. Voucher: Gillet 18059 (EA).Crassularhodesica (Merxm.) Wickens & M.Bywater \u2013 Life form: Herb. Habitat: Forest shades especially on rocky grounds, 1200\u20132100 m. Voucher: Verdcourt 670 (EA).Crassulaschimperi Fisch. & C.A.Mey. \u2013 Life form: Herb. Habitat: Moist rocky places in grassland, 1050\u20133820 m. Voucher: Bogdan 4635 (EA).Kalanchoedensiflora Rolfe \u2013 Life form: Herb. Habitat: Upland forest margins and grassland, 1000\u20133000 m. Voucher: SK 0072 .Sedumcrassularia (Schwienf.) R.-Hamet \u2013 Life form: Herb. Habitat: Rocky sites in moorland, 3300\u20134300 m. Voucher: Hedberg 1552 (EA).Sedummeyeri-johannis Engl. \u2013 Life form: Herb. Habitat: Epiphytic in moist forest and rocky heathland, 2100\u20133150 m. Voucher: SK 0265 .Sedumruwenzoriense Baker f. \u2013 Life form: Herb. Habitat: Rocky grounds in moorland, 2400\u20134500 m. Voucher: Mwangani 318 (EA).Umbilicus botryoides Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Epiphytic in wet montane forest, 2100\u20133900 m. Voucher: Kirika et al. 19 (EA).CucurbitaceaeF71. Cucumisficifolius A.Rich. \u2013 Life form: Herbaceous climber. Habitat: Grassland, 1070\u20132800 m. Voucher: Ekkens 661 (EA).Dactyliandrastefaninii (Chiov.) C.Jeffrey \u2013 Life form: Herbaceous climber. Habitat: Bushlands, 500\u20132594 m. Vouchers: SK 0036, SK 0071 .Diplocyclospalmatus (L.) C.Jeffrey \u2013 Life form: Herbaceous climber. Habitat: Moist forest and swampy grassland, 0\u20131830 m. Vouchers: SK 0220, SK 0066 .Lagenariaabyssinica (Hook.f.) C.Jeffrey \u2013 Life form: Herbaceous climber. Habitat: Upland moist forest and riparian forest, 900\u20133000 m. Voucher: SK 0230 .Momordicacalantha Gilg \u2013 Life form: Herbaceous climber. Habitat: Moist forest margins and valley grassland, 400\u20131900 m. Voucher: SK 0213 .Momordicafoetida Schumach. \u2013 Life form: Herbaceous climber. Habitat: Moist forest edges and open sites or glades, 1200\u20133000 m. Voucher: SK 0211 .Momordicafriesiorum (Harms) C.Jeffrey \u2013 Life form: Herbaceous climber. Habitat: Upland moist forest margins and glades, 1500\u20132850 m. Vouchers: SK 0211, SK 0214, SK 0232, SK 0252, SK 0218, SK 0188 .Oreosyceafricana Hook.f. \u2013 Life form: Herbaceous climber. Habitat: Moist forest margins and bamboo thickets, 900\u20133000 m. Voucher: SK 0144 .Peponiumvogelii (Hook.f.) Engl. \u2013 Life form: Herbaceous climber. Habitat: Moist forest and bamboo thickets, 10\u20132600 m. Voucher: Kamau 338 (EA).Zehneriaminutiflora (Cogn.) C.Jeffrey \u2013 Life form: Herbaceous climber. Habitat: Moist forest, 1100\u20133350 m. Voucher: Faden et al. 74/702 (EA).Zehneriascabra Sond. \u2013 Life form: Herbaceous climber. Habitat: Riverine forest and damp sites in bushland, 80\u20133350 m. Voucher: SAJIT 006501 (EA).Zehneriasubcoriaceae Y.D.Zhou & Q.F.Wang \u2013 Life form: Herbaceous climber. Habitat: Moist montane forest, 2000\u20133000 m. Voucher: SK 0137 .DichapetalaceaeF72. Dichapetalummadagascariensevar.brevistylum F.J.Breteler \u2013 Life form: Woody climber. Habitat: Upland evergreen forest, 1500\u20132400 m. Vouchers: Luke 384, Perdue and Kibuwa 8391 (EA).EbenaceaeF73. Diospyrosabyssinica (Hiern) F.White \u2013 Life form: Tree. Habitat: Montane bushland, 0\u20132400 m. Voucher: SK 0008 .Eucleadivinorum Hiern \u2013 Life form: Tree. Habitat: Grassland and open bushland, 0\u20132700 m. Voucher: Gardner 1392 (EA).Agaristasalicifolia (Lam.) G.Don \u2013 Life form: Shrub or tree. Habitat: Dry and moist forests, 1050\u20133500 m. Voucher: SK 0254 .EricaceaeF74. Ericaarborea L. \u2013 Life form: Shrub. Habitat: Upper montane forest and moorland, 1600\u20133900 m. Voucher: Gardner 1694 (EA).Ericamannii (Hook.f.) Beentje \u2013 Life form: Shrub. Habitat: Forest clearings in hilltops, 1200\u20132650 m. Vouchers: Mlawton 1803, Kuchar 10283 (EA).Ericasilvatica (Welw. ex Engl.) Beentje \u2013 Life form: Shrub. Habitat: Rocky grounds in moorland, 1650\u20134200 m. Voucher: Kokwaro 3242 (EA).Ericawhyteana Britten \u2013 Life form: Shrub. Habitat: Moist sites in moorland, 1900\u20133650 m. Voucher: Hedberg 1511 (EA).Ericafilago (Alm & T.C.E.Fr.) Beentje \u2013 Life form: Shrub. Habitat: Rocky sites in moorland, 2700\u20134350 m. Voucher: Kirika et al. 908 (EA).EuphorbiaceaeF75. Acalyphavolkensii Pax \u2013 Life form: Shrub. Habitat: Forest undergrowth and bushland, 800\u20133000 m. Voucher: SK 0192 .Brideliamicrantha (Hochst.) Baill. \u2013 Life form: Tree. Habitat: Evergreen forest and riparian forest, 0\u20132300 m. Voucher: SK 0034 .Clutiaabyssinicavar.abyssinica Jaub. & Spach \u2013 Life form: Shrub. Habitat: Upland bushland and wooded grassland, 1000\u20133700 m. Voucher: SK 0069 .Clutiaabyssinicavar.usambarica Pax & K.Hoffm \u2013 Life form: Shrub. Habitat: Dry evergreen forest, 300\u20132600 m. Voucher: Verdcourt 3135 (EA).Clutiakilimandscharica Engl. \u2013 Life form: Shrub. Habitat: Forest edges and bushland, 1700\u20133600 m. Voucher: Bally 13958 (EA).Crotonalienus Pax \u2013 Life form: Shrub or small tree. Habitat: Upland dry evergreen forest, 1525\u20131825 m. Voucher: Kamau 319 (EA).Crotonmacrostachyus Hochst. ex Delile \u2013 Life form: Tree. Habitat: Forest margins and along streams, 1350\u20132300 m. Voucher: SK 0033 .Crotonmegalocarpus Hutch. \u2013 Life form: Tree. Habitat: Wet and dry evergreen forest, 700\u20132400 m. Voucher: Kamau 319 (EA).Erythrococcabongensis Pax \u2013 Life form: Shrub. Habitat: Forest margins and bushland, 200\u20132440 m. Voucher: Kirika et al. 32 (EA).Euphorbiabrevicornu Pax \u2013 Life form: Herb. Habitat: Moist forest, 2000\u20133600 m. Voucher: Napier 657 (EA).Euphorbiabrevitorta P.R.O.Bally \u2013 Life form: Herb. Habitat: Dry bushland in rocky slopes, 1500\u20132000 m. Voucher: Kuchar 5105 (EA).Euphorbiadepauperata Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Rocky grounds in grassland and forest clearings, 1200\u20133350 m. Vouchers: Napier 655 (EA), SAJIT 006502 .Euphorbiaengleri Pax \u2013 Life form: Shrub. Habitat: Upland forest undergrowth and dense bushland, 1500\u20132800 m. Voucher: Hooper 1686 (EA).Euphorbiainaequilatera Sond. \u2013 Life form: Herb. Habitat: Swampy patches in upland grassland, 1990\u20132090 m. Voucher: Faden et al. 74/648 (EA).Euphorbiamagnicapsula S.Carter \u2013 Life form: Tree. Habitat: Open deciduous bushland, 1000\u20132165 m. Vouchers: Kirika & Muthoka 6, Perdue and Kibuwa 8264 (EA).Euphorbiascarlatina S.Carter \u2013 Life form: Shrub. Habitat: Open deciduous bushland, 600\u20132000 m. Voucher: Kirika 12 (EA).Euphorbiaschimperianavar.velutina N.E.Br. \u2013 Life form: Herb. Habitat: Open sites in montane forests and grassland, 2970\u20133760 m. Voucher: Hooper et al. 1684 (EA).Euphorbiaugandensis Pax & K.Hoffm. \u2013 Life form: Shrub. Habitat: Moist forest and bamboo thicket, 1980\u20133350 m. Voucher: Kerfoot 478 (EA).Euphorbiawellbyivar.wellbyi N.E.Br. \u2013 Life form: Shrub. Habitat: Moist upper parts montane forest zone and stream-sides in moorland, 3000\u20134000 m. Vouchers: Faden 74/856, Mabberley 333 (EA).Euphorbiawellbyivar.glabra S.Carter \u2013 Life form: Herb. Habitat: Upper montane forest edges and heathland, 2900\u20134000 m. Voucher: Beentje 2625 (EA).Euphorbiacandelabrum Tr\u00e9maux ex Kotschy \u2013 Life form: Tree. Habitat: Open wooded grassland, 900\u20132180 m. Voucher: Perdue and Kibuwa 8265 (EA).Heywoodialucens Sim \u2013 Life form: Tree. Habitat: Upland riparian forests, 1200\u20131950 m. Vouchers: JKCAT 1538, Seki (EA).Homalanthuspopulifolius Graham \u2013 Life form: Shrub to small tree. Habitat: Undergrowth in evergreen forests, 950\u20132100 m. Voucher: Gillett 20454 (EA).Macarangacapensis (Baill.) Sim \u2013 Life form: Tree. Habitat: Moist evergreen forest, 75\u20133050 m. Voucher: SK 0170 .Macarangakilimandscharica Pax \u2013 Life form: Tree. Habitat: Moist evergreen forest, 1310\u20133000 m. Voucher: Kuchar 5450 (EA).Micrococcaholstii (Pax) Prain \u2013 Life form: Shrub. Habitat: Moist evergreen forest, 1000\u20132400 m. Vouchers: Battiscombe 678, Faden & Evans 70/71 (EA).Neoboutoniamacrocalyx Pax \u2013 Life form: Tree. Habitat: Upland moist forest margins and forest clearings, 1100\u20132700 m. Voucher: Someren 3555 (EA).Phyllanthusboehmiivar.boehmii Pax \u2013 Life form: Herb. Habitat: Moist forest and woodland, 1050\u20133270 m. Vouchers: Kahurananga 2825, Verdcourt 400 (EA).Phyllanthusboehmiivar.humilis Radcl.-Sm. \u2013 Life form: Herb. Habitat: Damp sites in upland grassland and moorland, 2100\u20133250 m. Voucher: Napier 693 (EA).Phyllanthusfischeri Pax \u2013 Life form: Shrub. Habitat: Forest edges and along seasonal streams, 1450\u20132960 m. Voucher: Kamau 320 (EA).Tragiabrevipes Pax \u2013 Life form: Herb or subshrub. Habitat: Forest edges and riverine vegetation, 600\u20132600 m. Voucher: SK 0157 .Tragiellanatalensis (Sond.) Pax & K.Hoffm. \u2013 Life form: Herb. Habitat: Forest edges and associated bushland, 80\u20132300 m. Voucher: Verdcourt 546 (EA).Verniciafordii (Hemsl.) Airy Shaw \u2013 Life form: Exotic tree. Habitat: Cultivated along moist forest edges. Voucher: Patterson 324/58 (EA).*FabaceaeF76. Acaciaabyssinica Benth. \u2013 Life form: Tree. Habitat: Woodland and wooded grassland, 1500\u20132300 m. Voucher: Verdcourt 1501 (EA).Acaciaxanthophloea Benth. \u2013 Life form: Tree. Habitat: Riverine forest, 600\u20131980 m. Voucher: Ahiti 131 (EA).Adenocarpusmannii (Hook.f.) Hook.f. \u2013 Life form: Shrub. Habitat: Moorland, 1500\u20134000 m. Voucher: SK 0113 .Aeschynomeneschimperi A.Rich. \u2013 Life form: Shrub. Habitat: Swampy areas and along streams, 60\u20132340 m. Voucher: Battiscombe 193 (EA).Albiziagummifera (J.F.Gmel.) C.A.Sm. \u2013 Life form: Tree. Habitat: Moist forest, 0\u20132440 m. Voucher: Poster (EA).Amphicarpaeaafricana (Hook.f.) Harms \u2013 Life form: Herbaceous climber. Habitat: Upland moist forest and bamboo zone, 1680\u20132700 m. Voucher: Battiscombe 298 (EA).Argyrolobiumfriesianum (Hook.f.) Harms \u2013 Life form: Herb or subshrub. Habitat: Margins of upland moist forest, 1800\u20133000 m. Voucher: Mbale et al. 847 (EA).Argyrolobiumrupestresubsp.aberdaricum (Harms) Polhill \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 1900\u20133500 m. Voucher: Bally 2756 (EA).Argyrolobiumrupestresubsp.kilimandscharicum (Taub.) Polhill \u2013 Life form: Herb. Habitat: Upland grassland, 2250\u20133700 m. Voucher: Sir Charles 10607 (EA).Astragalusatropilosulusvar.astropilosulus (Hochst.) Bunge \u2013 Life form: Herb. Habitat: Upland grassland and forest margins, 1200\u20134200 m. Voucher: Beentje 2452 (EA).Astragalusatropilosulussubsp.bequaertii (De Wild.) J.B.Gillett \u2013 Life form: Herb. Habitat: Roadsides in grassland, 1200\u20134200 m. Voucher: Kerfoot 402 (EA).Astragalusatropilosulussubsp.burkeanus (Harvey) J.B.Gillett \u2013 Life form: Herb. Habitat: Bushland margins and open sites in bamboo forest, 2100\u20132700 m. Voucher: Verdcourt 691 (EA).Caesalpiniadecapetala (Roth) Alston \u2013 Life form: Exotic woody climber. Habitat: Open sites in montane forest and bushland, 880\u20132200 m. Voucher: Ament et al. 121 (EA).*Calpurniaaurea (Aiton) Benth. \u2013 Life form: Tree. Habitat: Upland rainforest margins and riverine forest, 1300\u20132260 m. Voucher: Kirika et al. 881 (EA).Chamaecristahildebrandtii (Vatke) Lock \u2013 Life form: Woody herb. Habitat: Wooded grassland, 1470\u20132300 m. Voucher: Faden 67419 (EA).Chamaecristastricta E.Mey. \u2013 Life form: Woody herb. Habitat: Roadsides in open bushland, 880\u20132040 m. Voucher: Whyte (EA).Chamaecristausambarensis (Taub.) Standl. \u2013 Life form: Herb. Habitat: Rocky grounds in upland grassland, 1760\u20132590 m. Voucher: Yyne-Watt 1187 (EA).Crotalariaagatiflorasubsp.agatiflora Schweinf. \u2013 Life form: Herb. Habitat: Upland grassland and deciduous bushland, 1500\u20133150 m. Voucher: SK 0051 .Crotalariaagatiflorasubsp.engleri (Baker f.) Polhill. \u2013 Life form: Herb. Habitat: Upland forest margins and riverine forest, 1500\u20133500 m. Voucher: Kimani 16 (EA).Crotalariaaxillaris Aiton \u2013 Life form: Shrub. Habitat: Forest margins and deciduous woodland, 0\u20132500 m. Voucher: Perdue 8064 (EA).Crotalariabrevidensvar.parviflora (Baker f.) Polhill \u2013 Life form: Herb. Habitat: Upland dry evergreen forest, 1500\u20133000 m. Vouchers: Verdcourt 2920, Strange 286 (EA).Crotalariafascicularis Polhill \u2013 Life form: Shrub. Habitat: Margins of upland rainforest, 1950\u20132950 m. Voucher: Gilbert 4896 (EA).Crotalariaincanavar.purpurascens (Lam.) Milne-Redh. \u2013 Life form: Herb. Habitat: Upland grassland, 1050\u20132600 m. Vouchers: Kokwaro 346, Kuchar 12255 (EA).Crotalariajacksonii Baker f. \u2013 Life form: Shrub. Habitat: Upland grassland and margins of moist forest, 2200\u20133000 m. Voucher: Kerfoot 4590 (EA).Crotalariakeniensis Baker f. \u2013 Life form: Shrub. Habitat: Moist forest margins and clearings, 1500\u20132850 m. Voucher: Becky 2175 (EA).Crotalarialebrunii Baker f. \u2013 Life form: Shrub. Habitat: Moist forest margins and clearings, 1350\u20132760 m. Voucher: Mathenge 206 (EA).Crotalariamauensis Baker f. \u2013 Life form: Shrub. Habitat: Upland forest margins, 1600\u20132800 m. Voucher: Verdcourt 681 (EA).Crotalarianatalitia Meissner \u2013 Life form: Woody herb or shrub. Habitat: Riverine forest and upland moist forest, 0\u20133000 m. Voucher: Kerfoot 481 (EA).Crotalariaprittwitzii Baker f. \u2013 Life form: Woody herb or shrub. Habitat: Riverine forest and upland moist forest, 0\u20133000 m. Voucher: Napper 637 (EA).Crotalariapseudospartium Baker f. \u2013 Life form: Shrub. Habitat: Upland wooded grassland, 1400\u20132500 m. Voucher: Verdcourt 3571 (EA).Crotalariarhizoclada Polhill \u2013 Life form: Shrub. Habitat: Roadsides in upland grassland, 1200\u20132500 m. Voucher: Lacey (EA).Crotalariatabularis Baker f. \u2013 Life form: Shrub. Habitat: Margins of upland rainforest, 1200\u20133000 m. Voucher: Mainwaring 2407 (EA).Dolichossericeussubsp.glabrescens Verdc. \u2013 Life form: Herb. Habitat: Upland bushland and dry evergreen forest, 1200\u20132780 m. Voucher: SK 0225 .Dolichossericeussubsp.pseudofalcatus Verdc. \u2013 Life form: Herb. Habitat: Upland grassland, 1200\u20132780 m. Voucher: Someren 1151 (EA).Eriosemascioanumsubsp.lejeunei (Staner & De Craene) Verdc. \u2013 Life form: Herb. Habitat: Upland forest glades and grassland, 1500\u20132580 m. Voucher: Verdcourt 1605 (EA).Erythrinaabyssinica DC. \u2013 Life form: Tree. Habitat: Scattered-tree grassland, 200\u20132100 m. Vouchers: Kirika 163, 157 (EA).Hylodesmumrepandum (Vahl) H.Ohashi & R.R.Mill \u2013 Life form: Herb. Habitat: Shaded grounds in upland moist forest, 1000\u20133000 m. Voucher: Kerfoot 473 (EA).Indigastrumcostatum (Guill. & Perr.) Schrire \u2013 Life form: Herb. Habitat: Short grassland, 500\u20131900 m. Voucher: Bally 955 (EA).Indigoferaarrecta A.Rich. \u2013 Life form: Woody herb. Habitat: Bushland and forest edges, 300\u20132700 m. Voucher: Kerfoot 392 (EA).Indigoferaatriceps Hook.f. \u2013 Life form: Herb. Habitat: Upland grassland and forest margins, 1000\u20133200 m. Vouchers: Kerfoot 1443 (EA), SK 0041 .Indigoferacircinella Baker f. \u2013 Life form: Woody herb. Habitat: Grassland, 50\u20132200 m. Voucher: Napier 1794 (EA).Indigoferademissa Taub. \u2013 Life form: Herb. Habitat: Roadsides in grassland, 900\u20132500 m. Voucher: Nattras 1285 (EA).Indigoferanairobiensissubsp.nairobiensis Baker f. \u2013 Life form: Woody herb. Habitat: Upland grassland, 1900\u20132300 m. Voucher: Bogdan 4624 (EA).Indigoferanairobiensissubsp.vicida J.B.Gillett \u2013 Life form: Woody herb. Habitat: Upland grassland, 1900\u20132300 m. Voucher: Gillet 19364 (EA).Indigoferaswaziensis Bolus \u2013 Life form: Shrub. Habitat: Upland evergreen forest margins, 1200\u20132700 m. Voucher: Hansen 762 (EA).Indigoferatritasubsp.scabra (Roth) Ali \u2013 Life form: Woody herb. Habitat: Grassland and bushland, 0\u20132500 m. Voucher: Malombe et al. 1381 (EA).Kotschyarecurvifoliasubsp.keniensis Verdc. \u2013 Life form: Shrub. Habitat: Dry evergreen bushland, 2340\u20133000 m. Vouchers: Gardner 1135, Dale 2684 (EA).Lathyrushygrophilus Taub. \u2013 Life form: Herbaceous climber. Habitat: Wet sites in moorland and bamboo forest, 1800\u20134100 m. Voucher: Mabberley 377 (EA).Lotusbecquetii Boutique \u2013 Life form: Herb. Habitat: Upland grassland, 2000\u20133200 m. Voucher: Hawery 168B (EA).Lotuscorniculatus L. \u2013 Life form: Herb. Habitat: Wet sites in upland grassland, 1400\u20132700 m. Voucher: Allnechtsen 10 (EA).Lotusgoetzei Harms \u2013 Life form: Herb. Habitat: Upland forest edges and grassland, 1500\u20133700 m. Voucher: Napier 689 (EA).Medicagolupulina L. \u2013 Life form: Herb. Habitat: Upland grassland, 1800\u20132900 m. Voucher: Kirika et al. 77 (EA).Melilotusofficinalis (L.) Pall. \u2013 Life form: Herb. Habitat: Roadsides in grassland, 2000\u20132000 m. Voucher: Kulkarni 14116 (EA).Neonotoniawightii (Wight & Arn.) J.A.Lackey \u2013 Life form: Woody climber. Habitat: Grassland and bushland, 0\u20132500 m. Voucher: Fries 537 (EA).Ormocarpumtrachycarpum (Taub.) Harms \u2013 Life form: Shrub or small tree. Habitat: Grassland and woodland, 950\u20131800 m. Voucher: Perdue and Kibuwa 8263 (EA).Otholobiumfoliosum (Oliv.) C.H.Stirt. \u2013 Life form: Shrub. Habitat: Upland grassland and forest margins, 1200\u20133200 m. Voucher: Fries 1581 (EA).Parochetuscommunis D.Don \u2013 Life form: Herb. Habitat: Upland moist forest and bamboo forest, 1500\u20133450 m. Voucher: Kuchar and Msafiri 5442 (EA).Rhynchosiacongensissubsp.orientalis Verdc. \u2013 Life form: Herb. Habitat: Grassland, 45\u20132280 m. Voucher: SK 0022 .Rhynchosiadensiflorasubsp.stuhlmannii (Harms) Verdc. \u2013 Life form: Herbaceous climber. Habitat: Upland grassland with scattered trees, 1200\u20132160 m. Vouchers: Nattrass 375, 588 (EA).Rhynchosiahirta (Andrews) Meikle & Verdc. \u2013 Life form: Herb. Habitat: Grassland and forest edges, 0\u20131850 m. Voucher: KEFRI & Omondi 106 (EA).Rhynchosiaminimavar.prostrata (Harv.) Meikle. \u2013 Life form: Herb. Habitat: Grassland, 45\u20132280 m. Vouchers: Verdcourt 1606, Agnew 7685 (EA).Rhynchosiausambarensissubsp.inelegans Verdc. Life form: Woody herb. Habitat: Upland forest edges and bushland, 1200\u20132400 m. Vouchers: Napier 2454, Robertson 1545 (EA).Sennadidymobotrya (Fresen.) H.S.Irwin & Barneby \u2013 Life form: Shrub. Habitat: Moist forest edges and riverine forest, 1500\u20132250 m. Voucher: SK 0209 .Sennaseptemtrionalis (Viv.) H.S.Irwin & Barneby \u2013 Life form: Exotic shrub. Habitat: Dry or moist forest, 910\u20133200 m. Vouchers: Mainwaring 2192 (EA) SK 0030 .*Sennasingueana (Delile) Lock \u2013 Life form: Shrub or small tree. Habitat: Wooded grassland, 0\u20132130 m. Voucher: Perdue and Kibuwa 8216 (EA).Stylosanthesfruticosa (Retz.) Alston \u2013 Life form: Herb or subshrub. Habitat: Grassland and bushland, 10\u20132720 m. Voucher: Perdue and Kibuwa 8183 (EA).Trifoliumburchellianum Ser. \u2013 Life form: Herb. Habitat: Clearings in upland forest and bamboo thickets, 1600\u20133980 m. Voucher: SK 0063 .Trifoliumcryptopodium Steud. ex A.Rich. \u2013 Life form: Herb. Habitat: Moist forest clearings and moorland, 1800\u20134200 m. Voucher: Gillett 19289 (EA).Trifoliumlanceolatum (J.B.Gillett) J.B.Gillett \u2013 Life form: Herb. Habitat: Upland grassland, 1950\u20132800 m. Voucher: Achlactoe 2749 (EA).Trifoliummultinerve A.Rich. \u2013 Life form: Herb. Habitat: Moist upland grassland and moorland, 1800\u20133700 m. Voucher: Mabberley 352 (EA).Trifoliumpolystachyum Fresen. \u2013 Life form: Herb. Habitat: Moist forest margins, 1600\u20132800 m. Voucher: Meinertzhagen (EA).Trifoliumsemipilosumvar.semipilosum Fresen. \u2013 Life form: Herb. Habitat: Upland grassland, 1500\u20133000 m. Voucher: Beentje 320g (EA).Trifoliumsemipilosumvar.glabrescens J.B.Gillett \u2013 Life form: Herb. Habitat: Upland grassland, 1200\u20132700 m. Vouchers: Trapnell 2111, Scott Elliot 6606 (EA).Trifoliumsimense Fresen. \u2013 Life form: Herb. Habitat: Upland grassland, 1500\u20133100 m. Voucher: Muasya et al. 002 (EA).Trifoliumsteudneri Schweinf. \u2013 Life form: Herb. Habitat: Upland grassland and forest margins, 1800\u20132400 m. Voucher: Bogdan 3222 (EA).Trifoliumtembense Fresen. \u2013 Life form: Herb. Habitat: Wet places in upland forest and moorland, 2000\u20133800 m. Voucher: Polhill 438 (EA).Viciabenghalensis L. \u2013 Life form: Exotic herb. Habitat: Disturbed areas in upland forest, 2400\u20132800 m. Voucher: Agriculture Dept 11568 (EA).*Viciahirsuta (L.) Gray \u2013 Life form: Herb. Habitat: Upland forest glades and grassland, 1950\u20133360 m. Voucher: Bogdan 1985 (EA).Viciasativa L. \u2013 Life form: Herb. Habitat: Upland grassland, 1700\u20133350 m. Voucher: Mabberley 376 (EA).Viciavillosasubsp.varia (Host) Corb. \u2013 Life form: Exotic herb. Habitat: Upland grassland, 1860\u20132700 m. Vouchers: Agric. Dept 11570, Blacklands 16293 (EA).*Vignaluteola (Jacq.) Benth. \u2013 Life form: Herbaceous climber. Habitat: Swampy forest and wet grassland, 650\u20131920 m. Vouchers: Battiscombe 1123, Kirrika 230, Napper 412 (EA).Vignamembranaceasubsp.macrodon (Robyns & Boutique) Verdc. \u2013 Life form: Herbaceous climber. Habitat: Upland moist forest, 1275\u20132100 m. Vouchers: Verdcourt 661, EANHS KF/77 (EA).Vignaparkeri Baker \u2013 Life form: Herbaceous climber. Habitat: Upland grassland with scattered trees, 1050\u20132900 m. Voucher: Kerfoot 1369 (EA).GentianaceaeF77. Sebaeabrachyphylla Griseb. \u2013 Life form: Herb. Habitat: Moist montane forests and moorlands, 1400\u20133470 m. Voucher: SK 0118 .Sebaealeiostyla Gilg \u2013 Life form: Herb. Habitat: Stream-sides and marshes in upland grassland, 1800\u20133500 m. Voucher: Kerfoot 1369 (EA).Sebaeapentandravar.burchellii E.Mey. \u2013 Life form: Herb. Habitat: Stream-sides and marshes in upland grassland, 1450\u20132450 m. Voucher: Greenway 13569 (EA).Swertiacrassiusculavar.crassiuscula Gilg \u2013 Life form: Herb. Habitat: Moorland, 2700\u20134500 m. Vouchers: Copley 138 (EA), SK 0087 .Swertiacrassiusculavar.leucantha (T.C.E.Fr.) Sileshi \u2013 Life form: Herb. Habitat: Moorland, 2700\u20134500 m. Voucher: Dowson 101 (EA).Swertiaeminii Engl. \u2013 Life form: Herb. Habitat: Wet grassland and swamp margins, 1200\u20132250 m. Voucher: Paulo et al. 888 (EA).Swertiakilimandscharica Engl. \u2013 Life form: Herb. Habitat: Open sites in montane forest, 2100\u20133840 m. Voucher: Bally 8643 (EA).Swertialugardiae Bullock \u2013 Life form: Herb. Habitat: Montane grassland, 2450\u20133550 m. Voucher: Naper 1231 (EA).Swertiavolkensii Gilg \u2013 Life form: Herb. Habitat: Upper parts of montane forest to the moorland, 2800\u20134250 m. Voucher: Hedberg 1544 (EA).GeraniaceaeF78. Geraniumaculeolatum Oliv. \u2013 Life form: Herb. Habitat: Upland rainforest, 1200\u20133400 m. Voucher: Mathenge 220 (EA).Geraniumarabicumsubsp.arabicum Forssk. \u2013 Life form: Herb. Habitat: Rainforest and moist sites in grassland and moorland, 1100\u20133940 m. Voucher: SAJIT 006498 .Geraniumarabicumsubsp.latistipulatum (Hochst. ex A.Rich.) Kokwaro \u2013 Life form: Herb. Habitat: Upland grassland and bushland, 1100\u20132800 m. Vouchers: Mwangangi 989, Kokwaro 32 (EA).Geraniumkilimandscharicum Engl. \u2013 Life form: Herb. Habitat: Rocky sites in moorland, 2260\u20134300 m. Voucher: SK 0059 .Geraniummascatense Boiss. \u2013 Life form: Herb. Habitat: Upland grassland and evergreen bushland, 1000\u20132900 m. Voucher: Verdcourt 679 (EA).Geraniumocellatum Cambess. \u2013 Life form: Herb. Habitat: Upland wooded grassland and evergreen bushland, 1000\u20132900 m. Voucher: Verdcourt 679 (EA).Geraniumpurpureum Vill. \u2013 Life form: Herb. Habitat: Upland rainforest and riverine forest, 1300\u20133000 m. Vouchers: Napier 729, Pierce 1682 (EA).Geraniumvaganssubsp.vagans Baker \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 1370\u20134500 m. Vouchers: SAJIT 006478, SK 0108 .Geraniumvaganssubsp.whytei (Baker) J.R.Laundon \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 1370\u20134500 m. Voucher: Kuchar 12495 (EA).Pelargoniumalchemilloides (L.) Aiton \u2013 Life form: Herb. Habitat: Bushland and montane forest edges, 700\u20132800 m. Voucher: Townsend 2319 (EA).Pelargoniuminquinans (L.) L\u2019Her. \u2013 Life form: Herb. Habitat: Bushland and wooded grassland, 700\u20132800 m. Voucher: SK 0269 .GesneriaceaeF79. Streptocarpusglandulosissimus Engl. \u2013 Life form: Herb. Habitat: Moist forest, 900\u20132600 m. Voucher: Bally 8517 (EA).GunneraceaeF80. Gunneraperpensa L. \u2013 Life form: Herb. Habitat: Upland riparian forest, 1560\u20134000 m. Voucher: Gardner 1882 (EA).HamamelidaceaeF81. Trichocladusellipticus Eckl. & Zeyh. \u2013 Life form: Tree. Habitat: Upland moist forest, 1350\u20132800 m. Voucher: Hansen 808 (EA).HypericaceaeF82. Hypericumkiboense Oliv. \u2013 Life form: Shrub. Habitat: Upland dry evergreen forests and grassland, 2100\u20133900 m. Voucher: Battiscombe 710 (EA).Hypericumlalandii Choisy \u2013 Life form: Herb. Habitat: Marshes and damp sites in upland grassland, 1080\u20132250 m. Voucher: Taylor 1511 (EA).Hypericumlanceolatum Lam. \u2013 Life form: Shrub or small tree. Habitat: Upland dry evergreen forests, 1800\u20133360 m. Voucher: Edwards 2843/16 (EA).Hypericumpeplidifolium A.Rich. \u2013 Life form: Herb. Habitat: Wet places in moorland, 1170\u20133600 m. Voucher: Coe 788 (EA).Hypericumrevolutumsubsp.revolutum Vahl \u2013 Life form: Shrub. Habitat: Upland dry evergreen forest, 2100\u20133250 m. Vouchers: SAJIT 006484, SK 0117 .Hypericumrevolutumsubsp.keniense (Schweinf.) N.Robson \u2013 Life form: Shrub. Habitat: Dry evergreen forest, 2700\u20133800 m. Voucher: Knox 3722 (EA).Hypericumscioanum Chiov. \u2013 Life form: Herb. Habitat: Wet sites in moorland, 1830\u20133590 m. Voucher: Mabberley 344 (EA).IcacinaceaeF83. Apodytesdimidiata E.Mey. ex Arn. \u2013 Life form: Tree. Habitat: Upland moist forest, 1000\u20132500 m. Voucher: SK 0146 .LamiaceaeF84. Achyrospermumschimperi (Hochst. ex Briq.) Perkins \u2013 Life form: Herb. Habitat: Montane forest undergrowth, 1200\u20133000 m. Voucher: SK 0208 .Ajugaintegrifolia Buch.-Ham. ex D.Don \u2013 Life form: Herb. Habitat: Upland moist forest, 1000\u20133400 m. Voucher: SK 0197 .Clerodendrumjohnstonii Oliv. \u2013 Life form: Woody climber. Habitat: Upland moist forest, 1200\u20132550 m. Voucher: Verdcourt 3780 (EA).Clinopodiumabyssinicumvar.condensatum (Hedberg) Ryding \u2013 Life form: Shrub. Habitat: Wet evergreen bushland and grassland, 1000\u20133950 m. Vouchers: Lind 2934, Agricultural Dept 62 (EA).Clinopodiumkilimandschari (G\u00fcrke) Ryding \u2013 Life form: Herb. Habitat: Moorland and heath zone, 2900\u20134400 m. Voucher: SK 0090 .Clinopodiumsimense (Benth.) Kuntze \u2013 Life form: Herb. Habitat: Moist grassland and open woodland, 1700\u20133500 m. Voucher: Fries and Fries 2429 (EA).Clinopodiumuhligii (G\u00fcrke) Ryding \u2013 Life form: Shrub. Habitat: Montane forest edges and evergreen bushland, 3350\u20134180 m. Voucher: Kuchar 10360 (EA).Fuerstiaafricana T.C.E.Fr. \u2013 Life form: Woody herb. Habitat: Upland grassland, 1200\u20132550 m. Voucher: Kuchar 8357a (EA).Leonotisnepetifolia (L.) R.Br. \u2013 Life form: Herb. Habitat: Upland grassland and bushland, 1000\u20132290 m. Voucher: Napier 2587 (EA).Leonotisocymifoliavar.raineriana (Vis.) Iwarsson. \u2013 Life form: Shrub. Habitat: Margins of montane forest, 600\u20133700 m. Vouchers: Verdcourt 3813 (EA), SK 0097 .Leucasdeflexa Hook.f. \u2013 Life form: Herb. Habitat: Moist forest, 1000\u20132500 m. Voucher: Gilbert 6312 (EA).Leucasgrandis Vatke \u2013 Life form: Herb or subshrub. Habitat: Upland evergreen forest, 500\u20132780 m. Voucher: Kamau 367 (EA).Leucasmasaiensis Oliv. \u2013 Life form: Herb. Habitat: Upland forest glades and margins, 1300\u20133200 m. Voucher: Kirika et al. 163 (EA).Leucasmasaiensisvar.venulosa (Baker) Sebald \u2013 Life form: Herb. Habitat: Upland grassland, 1300\u20133200 m. Voucher: Faden et al. 74/577 (EA).Leucasoligocephalavar.oligocephala Hook.f. Life form: Herb. Habitat: Upland grassland and forest margins, 1600\u20132990 m. Vouchers: Verdcourt 1010, 1023 (EA).Leucasvolkensiivar.parviflora Sebald. \u2013 Life form: Herb. Habitat: Open sites in montane forest, 2000\u20132600 m. Vouchers: Taylor 1237, Beentje 3254 (EA).Menthaaquatica L. \u2013 Life form: Herb. Habitat: Upland marshes, 1100\u20132150 m. Voucher: Kuchar 9573 (EA).Menthalongifolia (L.) L. \u2013 Life form: Herb. Habitat: Upland grassland and marshes, 1650\u20132450 m. Vouchers: MacDonald 1344, Poster 3234 (EA).Micromeriaimbricatavar.imbricata (Forssk.) C.Chr. \u2013 Life form: Woody herb. Habitat: Upland open woodland and dry grassland, 1200\u20134000 m. Voucher: Napier 1761 (EA).Micromeriaimbricatavar.villosa Ryding \u2013 Life form: Woody herb. Habitat: Upland evergreen bushland and grassland, 1850\u20134100 m. Voucher: Dawson 415 (EA).Nepetaazurea R.Br. ex Benth. \u2013 Life form: Herb. Habitat: Upland evergreen bushland and grassland, 1700\u20133800 m. Voucher: SK 0120 .Ocimumdecumbens G\u00fcrke \u2013 Life form: Shrub. Habitat: Grassland, 950\u20134000 m. Voucher: Lind 3138 (EA).Ocimumgratissimum L. \u2013 Life form: Herb. Habitat: Dry montane forest, 1100\u20132400 m. Voucher: Perdue and Kibuwa 8046 (EA).Ocimumkenyense Ayob. ex A.J.Paton \u2013 Life form: Herb. Habitat: Wet places in grassland, 1050\u20132300 m. Voucher: Mainwaring 2406 (EA).Ocimumkilimandscharicum G\u00fcrke \u2013 Life form: Shrub. Habitat: Upland grassland, 1100\u20132350 m. Voucher: Faden 68/721 (EA).Ocimumlamiifolium Hochst. ex Benth. \u2013 Life form: Shrub. Habitat: Upland forest edges and bushland, 1000\u20132500 m. Voucher: Kokwaro et al. 2344 (EA).Platostomadenticulatum Robyns \u2013 Life form: Herb. Habitat: Damp sites in grassland and open woodland, 100\u20132480 m. Voucher: Kerfoot 639 (EA).Plectranthusalboviolaceus G\u00fcrke \u2013 Life form: Shrub. Habitat: Upland moist forest and along rivers, 1500\u20132800 m. Voucher: Kimani 22 (EA).Plectranthusalpinus (Vatke) Ryding \u2013 Life form: Shrub. Habitat: Moist forest and along streams, 1900\u20132750 m. Voucher: Napper 657 (EA).Plectranthuscaespitosus Lukhoba & A.J.Paton \u2013 Life form: Herb. Habitat: Upland grassland, 1500\u20132850 m. Voucher: Someren 28 (EA).Plectranthuskamerunensis G\u00fcrke \u2013 Life form: Herb. Habitat: Glades in moist forest and bamboo zone, 1200\u20132700 m. Voucher: Young 1021 (EA).Plectranthuslaxiflorus Benth. \u2013 Life form: Herb. Habitat: Glades in moist forest and bamboo forest, 1600\u20133120 m. Voucher: Alexander 11636 (EA).Plectranthuslongipes Baker \u2013 Life form: Herb or subshrub. Habitat: Rocky grounds in bushland and woodland, 700\u20132440 m. Voucher: Patel 164 (EA).Plectranthusmelleri Baker \u2013 Life form: Shrub. Habitat: Open areas in moist montane forest, 1300\u20132400 m. Voucher: Kokwaro and Mathenge 2981 (EA).Plectranthusmollis (Aiton) Spreng. \u2013 Life form: Herb. Habitat: Upland grassland and forest margins, 1200\u20132900 m. Voucher: SK 0231 .Plectranthusmontanus Benth. \u2013 Life form: Herb. Habitat: Dry evergreen forest, 500\u20132400 m. Voucher: Bally 2602 (EA).Plectranthusparvus Oliv. \u2013 Life form: Herb. Habitat: Upland grassland and forest margins, 1200\u20132900 m. Voucher: Faden 67/838 (EA).Plectranthuspunctatussubsp.edulis (Vatke) A.J.Paton \u2013 Life form: Herb. Habitat: Moist montane forest up to bamboo zone, 1800\u20133200 m. Vouchers: Kerfoot 51, Mwangangi 977 (EA).Plectranthussylvestris G\u00fcrke \u2013 Life form: Shrub. Habitat: Moist montane forest up to bamboo zone, 1750\u20133280 m. Voucher: Agnew 7705 (EA).Rothecamyricoidesvar.myricoides (Hochst.) Steane & Mabb. \u2013 Life form: Shrub. Habitat: Grassland and open woodland, 900\u20132400 m. Voucher: Faden 74/570 (EA).Rothecamyricoidesvar.discolor (Klotzsch) Verdc. \u2013 Life form: Shrub. Habitat: Grassland and open woodland, 900\u20132400 m. Voucher: Hansen 781 (EA).Salviamerjamie Forssk. \u2013 Life form: Herb. Habitat: Montane grassland and moorland, 2250\u20134100 m. Voucher: Mbale et al. 861 (EA).Salvianilotica Juss. ex Jacq. \u2013 Life form: Exotic herb. Habitat: Montane grassland and forest edges, 1350\u20133700 m. Voucher: Harvey 179 (EA).*Stachysaculeolata Hook.f. \u2013 Life form: Herb. Habitat: Moist sites in bamboo zone and moorland, 1400\u20133650 m. Voucher: Napier 698 (EA).Stachysalpigena T.C.E.Fr. \u2013 Life form: Herb. Habitat: Montane moorland and ericaceous zone, 2900\u20133750 m. Vouchers: Kokwaro 1908 (EA), SK 0064 .Stachysargillicola Sebsebe \u2013 Life form: Woody herb. Habitat: Upland grassland, 1660\u20132200 m. Voucher: Evans 60/167 (EA).Tinneaaethiopica Kotschy ex Hook.f. \u2013 Life form: Herb. Habitat: Wooded grassland, 0\u20132300 m. Voucher: Faden 74/578 (EA).Vitexkeniensis Turrill \u2013 Life form: Tree. Habitat: Moist evergreen forest, 1290\u20132100 m. Voucher: SK 0248 .LauraceaeF85. Ocoteakenyensis (Chiov.) Robyns & R.Wilczek \u2013 Life form: Tree. Habitat: Moist montane forest, 1140\u20132400 m. Voucher: Elliot 2356 (EA).Ocoteausambarensis Engl. \u2013 Life form: Tree. Habitat: Moist montane forest, 900\u20133000 m. Voucher: Forest Dept. 176 (EA).LentibulariaceaeF86. Utriculariagibba L. \u2013 Life form: Herb. Habitat: Aquatic in shallow flowing water and freshwater pools, 10\u20132550 m. Vouchers: William 12346 & 12347 (EA).Utricularialivida E.Mey. \u2013 Life form: Herb. Habitat: Wet grassland, 0\u20132730 m. Voucher: Gilbert 4868 (EA).LinaceaeF87. Linumkeniense T.C.E.Fr. \u2013 Life form: Herb. Habitat: Upland grassland and open grounds in bamboo thickets, 2200\u20133360 m. Vouchers: SAJIT 006471 & 006489 .Linumvolkensii Engl. \u2013 Life form: Herb. Habitat: Wet grassland and stream banks, 1300\u20132750 m. Voucher: Symes 131 (EA).LoganiaceaeF88. Buddlejapolystachya Fresen. \u2013 Life form: Shrub. Habitat: Margins and clearings in upland rainforest, 1000\u20132700 m. Voucher: Birch 61/31 (EA).Nuxiacongesta R.Br. ex Fresen \u2013 Life form: Tree. Habitat: Upland rainforest, 1550\u20132850 m. Voucher: Kokwaro 4421 (EA).LoranthaceaeF89. Agelanthusbrunneus (Engl.) Balle & N.Hall\u00e9 \u2013 Life form: Shrub. Habitat: Moist evergreen forest and riparian forest, 1000\u20131800 m. Voucher: Someren 3191 (EA).Agelanthuspennatulus (Sprague) Polhill & Wiens \u2013 Life form: Shrub. Habitat: Moist montane forest, 1650\u20132400 m. Voucher: Kuchar and Msafiri 5461 (EA).Agelanthussansibarensissubsp.montanus Polhill & Wiens \u2013 Life form: Shrub. Habitat: Moist montane forest, 0\u20132500 m. Vouchers: Wiens 4564, Mainwaring s.n. (EA).Agelanthussubulatus (Engl.) Polhill & Wiens \u2013 Life form: Shrub. Habitat: Bushland and wooded grassland, 10\u20132300 m. Voucher: Faden & Evans 74/706 (EA).Englerinawoodfordioides (Schweinf.) Balle \u2013 Life form: Shrub. Habitat: Moist montane forest and riverine forest, 1350\u20133050 m. Vouchers: SK 0070, SK 0240 .Oncocalyxsulfureus (Engl.) Wiens & Polhill -Life form: Herb. Habitat: Epiphytic in upland dry evergreen forest, 1700\u20133000 m. Voucher: Hepper 4916 (EA).LythraceaeF90. Parsonsiamicropetala (Kunth) Standl. \u2013 Life form: Exotic shrub. Habitat: Escaped cultivation common on stream-sides and disturbed sites. Voucher: Hooper and Townsend 1691 (EA).*Lythrumrotundifolium Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland water pools and swamps, 1650\u20133300 m. Voucher: Kahurananga et al. 2820 (EA).Nesaeakilimandscharicavar.ngongensis Verdc. \u2013 Life form: Woody herb or shrub. Habitat: Upland grassland and bushland, 1650\u20132130 m. Vouchers: Hansen 816, Gillett 17342 (EA).Nesaeaschinziisubsp.subalata (Koehne) Verdc. \u2013 Life form: Herb or subshrub. Habitat: Scattered-tree grassland, 1080\u20132100 m. Vouchers: Dowson 541, Dowson 541 (EA).MalvaceaeF91. Abutilonlongicuspevar.longicuspe Hochst. ex A.Rich. \u2013 Life form: Shrub. Habitat: Open sites in dry evergreen forest, 1650\u20133300 m. Voucher: Faden 67719 (EA).Abutilonlongicuspevar.pilosicalyx Verdc. \u2013 Life form: Shrub. Habitat: Upland grassland and dry forest margins, 1700\u20132400 m. Voucher: Dyson 456 (EA).Abutilonmauritianum (Jacq.) Medik. \u2013 Life form: Shrub. Habitat: Woodland and forest edges, 0\u20132300 m. Voucher: Kokwaro & Kabuye 329 (EA).Dombeyakirkii Mast. \u2013 Life form: Shrub or small tree. Habitat: Bushland and forest margins, 600\u20132400 m. Voucher: SK 0132 .Dombeyarotundifolia (Hochst.) Planch. \u2013 Life form: Tree. Habitat: Forest edges and wooded grassland, 1000\u20132400 m. Voucher: Someren 692 (EA).Dombeyatorrida (J.F.Gmel.) Bamps \u2013 Life form: Tree. Habitat: Upland open forests and forest margins, 1700\u20133050 m. Voucher: Brasnett 200 (EA).Grewiasimilis K.Schum. \u2013 Life form: Shrub. Habitat: Woodland and grassland, 600\u20132250 m. Voucher: Fukuoka 106 (EA).Hibiscusfuscus Garcke \u2013 Life form: Woody herb. Habitat: Upland bush thickets and grassland, 1400\u20132650 m. Voucher: Kokwaro & Kabuye 340 (EA).Hibiscusmacranthus Hochst. ex A.Rich. \u2013 Life form: Shrub. Habitat: Upland forest edges and bushland, 1500\u20132900 m. Voucher: Kuchar 12297 (EA).Hibiscusvitifoliussubsp.vitifolius Brenan & Exell \u2013 Life form: Shrub. Habitat: Dry bushland, 420\u20133000 m. Vouchers: Robertson 1805, McDonald 922 (EA).Malvaverticillata L. \u2013 Life form: Herb. Habitat: Moist montane forest, 1200\u20134050 m. Voucher: McDonald 1285 (EA).Pavoniaburchellii (DC.) R.A.Dyer \u2013 Life form: Shrub. Habitat: Woodland and rainforest margins, 750\u20132300 m. Voucher: SK 0203 .Pavoniaschimperiana Hochst. ex A.Rich. \u2013 Life form: Woody herb or shrub. Habitat: Upland short grassland and forest edges, 1100\u20132400 m. Voucher: Smith et al. 66 (EA).Pavoniaurens Cav. \u2013 Life form: Shrub. Habitat: Forest margins and riparian vegetation, 600\u20133000 m. Vouchers: SK 0020, SK 0068, SK 0261 .Sidacordifolia L. \u2013 Life form: Herb or subshrub. Habitat: Upland bushland, 1300\u20132291 m. Voucher: SK 0163 .Sidarhombifolia L. \u2013 Life form: Shrub. Habitat: Open woodland, 900\u20132250 m. Voucher: SK 0242 .Sidaschimperiana Hochst. ex A.Rich. \u2013 Life form: Shrub. Habitat: Upland grassland, 1200\u20132700 m. Voucher: Smith et al. 66 (EA).Sidatenuicarpa Vollesen \u2013 Life form: Shrub. Habitat: Roadsides in forest, 750\u20132400 m. Voucher: SK 0015 .Sidaternata L.f. \u2013 Life form: Herb. Habitat: Dry montane forest, 1350\u20133280 m. Voucher: Taylor 1586 (EA).Sparmanniaricinocarpa (Eckl. & Zeyh.) Kuntze \u2013 Life form: Shrub. Habitat: Upland grassland with moist forest edges, 1550\u20133380 m. Voucher: Blake 10858 (EA).Triumfettabrachyceras K.Schum. \u2013 Life form: Woody herb or shrub. Habitat: Moist forest clearings and margins, 1200\u20133000 m. Voucher: Musili et al. 187 (EA).Triumfettalongicornuta Hutch. & M.B.Moss \u2013 Life form: Shrub. Habitat: Glades in dry evergreen forest, 1350\u20132150 m. Voucher: SK 0125 .Triumfettapilosa Roth \u2013 Life form: Shrub. Habitat: Moist forest and swamp edges, 1200\u20132250 m. Voucher: Stroud 78 6732 (EA).MeliaceaeF92. Ekebergiacapensis Sparrm. \u2013 Life form: Tree. Habitat: Montane forest and riparian forest, 600\u20132750 m. Voucher: Gardner 380 (EA).Lepidotrichiliavolkensii (G\u00fcrke) Leroy \u2013 Life form: Tree. Habitat: Upland forest margins, 1550\u20132600 m. Voucher: Napper 1491 (EA).Trichiliadregeana Sond. \u2013 Life form: Tree. Habitat: Moist forest and riparian forest, 775\u20131800 m. Voucher: Gachathi 2/81 (EA).Turraeaabyssinica Hochst. ex A.Rich. \u2013 Life form: Shrub or small tree. Habitat: Montane forest, 1820\u20132225 m. Voucher: Gardner 542 (EA).Turraeamombassanasubsp.cuneata (G\u00fcrke) Styles & F.White \u2013 Life form: Shrub or small tree. Habitat: Upland dry forest and bushlands, 1525\u20132225 m. Vouchers: Faden 6742, Hansen 760 (EA).MelianthaceaeF93. Bersamaabyssinica Fresen. \u2013 Life form: Tree. Habitat: Upland moist forest, 1140\u20132550 m. Vouchers: Davidse 7058 (EA), SK 0013, SK 0139, SK 0233 .MenispermaceaeF94. Cissampelosfriesiorum Diels \u2013 Life form: Herbaceous climber. Habitat: Upland moist forest, 2000\u20132100 m. Voucher: Fries 1625 (EA).Stephaniaabyssinicavar.abyssinica (Quart.-Dill & A.Rich.) Walp. \u2013 Life form: Herbaceous climber. Habitat: Moist shaded sites in wooded grassland, 1450\u20133500 m. Vouchers: SAJIT 006465, SK 0035 .Stephaniaabyssinicavar.tomentella (Oliv.) Diels \u2013 Life form: Herbaceous climber. Habitat: Moist shaded sites in wooded grassland, 1450\u20133500 m. Voucher: SK 0159 .MoraceaeF95. Dorsteniaafromontana R.E.Fr. \u2013 Life form: Herb. Habitat: Upland rainforest, 2000\u20132600 m. Voucher: Kirika et al. 76 (EA).Dorsteniahildebrandtii Engl. \u2013 Life form: Herb. Habitat: Epiphytic in moist forest and stream banks, 300\u20132170 m. Voucher: Napier 2182 (EA).Ficuscordatasubsp.salicifolia (Vahl) C.C.Berg \u2013 Life form: Tree. Habitat: Riparian forest and seasonal streams, 950\u20132400 m. Vouchers: Verdcourt 3548, Makin 26 (EA).Ficussur Forssk. \u2013 Life form: Tree. Habitat: Riverine forest, 350\u20132500 m. Vouchers: SK 0228, SK 0229 .Ficusthonningii Blume \u2013 Life form: Tree. Habitat: Moist forest, 350\u20132500 m. Voucher: Kamau 310 (EA).MyricaceaeF96. Morellasalicifoliasubsp.meyeri-johannis (Engl.) Verdc. & Polhill. \u2013 Life form: Tree. Habitat: Upland grassland and moorland, 2700\u20133700 m. Vouchers: Beentje 3240, Thairu 16861 (EA).MyrtaceaeF97. Corymbiacalophylla (R.Br. ex Lindl.) K.D.Hill & L.A.S.Johnson \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Muasya 2020 (EA).*Corymbiagummifera (Gaertn.) K.D.Hill & L.A.S.Johnson \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Pudden 16 (EA).*Eucalyptuscrebra F.Muell. \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Greenway 8762 (EA).*Eucalyptusglobulussubsp.maidenii (F.Muell.) J.B.Kirkp. \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Vouchers: Verdcourt 12800, Forest Dept 16083 (EA).*Eucalyptuslongifolia Link & Otto. \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Forest Dept 16098 (EA).*Eucalyptusmicrocorys F.Muell. \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Forest Dept 16108 (EA).*Eucalyptusmuelleriana Howitt \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Pudden 26 (EA).*Eucalyptusobliqua L\u2019H\u00e9r. \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Pudden 054 (EA).*Eucalyptuspaniculata Sm. \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Stuart 2 (EA).*Eucalyptuspellita F.Muell. \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Greenway 8754 (EA).*Eucalyptuspunctata A.Cunn. ex DC. \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Darling 31 (EA).*Eucalyptussiderophloia Benth. \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Verdcourt 1963 (EA).*Eucalyptusviminalis Labill. \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides and moist forest edges. Voucher: Forest Dept. 16137 (EA).*Syzygiumguineensesubsp.afromontanum F.White \u2013 Life form: Exotic tree. Habitat: Upland riverine and moist forest, 1500\u20132550 m. Vouchers: Battiscombe 555, Moore 762 (EA).OchnaceaeF98. Ochnaholstii Engl. \u2013 Life form: Tree. Habitat: Upland moist forest, 900\u20132350 m. Voucher: Polhill 165 (EA).Ochnainsculpta Sleumer \u2013 Life form: Tree. Habitat: Evergreen forest, 1050\u20132450 m. Voucher: Agnew et al. 7939 (EA).OlacaceaeF99. Strombosiascheffleri Engl. \u2013 Life form: Tree. Habitat: Moist forest, 800\u20132500 m. Voucher: SK 0028 .OleaceaeF100. Chionanthusmildbraedii (Gilg & G.Schellenb.) Stearn \u2013 Life form: Tree. Habitat: Upland moist forest and riverine forest, 1200\u20132100 m. Voucher: SK 0238 .Fraxinuspennsylvanica Marshall \u2013 Life form: Exotic tree. Habitat: Cultivated in roadsides. Voucher: SK 0127 .*Jasminumabyssinicum Hochst. ex DC. \u2013 Life form: Woody climber. Habitat: Forest undergrowth and forest margins, 690\u20133000 m. Voucher: McDonald 1336 (EA).Jasminumschimperi Vatke \u2013 Life form: Woody climber. Habitat: Rainforest and wooded grassland, 690\u20133000 m. Voucher: Miss Mainwaring 2404 (EA).Oleaeuropaea L. \u2013 Life form: Tree. Habitat: Upland wet forest, 950\u20132400 m. Voucher: Napper 1697 (EA).Oleacapensis L. \u2013 Life form: Tree. Habitat: Upland dry forest, 1150\u20132680 m. Voucher: SK 0105 .OnagraceaeF101. Epilobiumhirsutum L. \u2013 Life form: Herb. Habitat: Damp sites in upland grassland and moorland, 1190\u20132590 m. Voucher: SK 0237 .Epilobiumstereophyllum Fresen. \u2013 Life form: Herb. Habitat: Damp sites in upland grassland and moorland, 1750\u20133500 m. Voucher: SK 0116 .Fuchsiaarborescens Sims \u2013 Life form: Exotic shrub or tree. Habitat: Moist forest, 1220\u20132490 m. Voucher: SK 0130 .*Ludwigiaadscendenssubsp.diffusa (Forssk.) P.H.Raven \u2013 Life form: Herb. Habitat: Swamps and freshwater pools, 600\u20131900 m. Voucher: SK 0161 .OrobanchaceaeF102. Hedbergiaabyssinica (Benth.) Molau \u2013 Life form: Herb. Habitat: Montane grassland and forest margins, 2000\u20133980 m. Voucher: Hedberg 1646 (EA).Orobancheminor Sm. \u2013 Life form: Herb. Habitat: Forest edges and disturbed grounds, 540\u20133000 m. Voucher: Townsend 2294 (EA).Orobancheramosa L. \u2013 Life form: Herb. Habitat: Roadsides in upland grassland and woodland, 1735\u20132250 m. Vouchers: The Wallis 15436, Scaham 22 (EA).OxalidaceaeF103. Oxaliscorniculata L. \u2013 Life form: Herb. Habitat: Forest glades, 0\u20133600 m. Voucher: SAJIT 006481 .PapaveraceaeF104. Corydaliscornuta Royle \u2013 Life form: Herb. Habitat: Montane forest up to the moorland, 2300\u20133300 m. Voucher: Napier 720 (EA).Corydalismildbraedii Fedde \u2013 Life form: Herb. Habitat: Montane forest up to moorland, 2200\u20133600 m. Vouchers: SAJIT 006463, SK 0176 .Fumariaabyssinica Hammar \u2013 Life form: Herb. Habitat: Montane forest up to moorland, 1300\u20133200 m. Voucher: Verdcourt 3207 (EA).PassifloraceaeF105. Adeniaglobosasubsp.pseudoglobosa (Verdc.) W.J. de Wilde \u2013 Life form: Woody climber. Habitat: Deciduous and dry evergreen forest, 0\u20131850 m. Vouchers: Verdcourt 2677, 3547 (EA).Adeniagummifera (Harv.) Harms \u2013 Life form: Woody climber. Habitat: Dry or moist forest and bushland, 0\u20131850 m. Voucher: SK 0195 .Passifloraedulis Sims \u2013 Life form: Exotic herbaceous climber. Habitat: Moist forest edges and bush thickets, 0\u20132500 m. Voucher: Greenway 10899 (EA).*Passifloramollissima L.H.Bailey \u2013 Life form: Exotic woody climber. Habitat: Moist forest edges, 1000\u20133000 m. Voucher: SK 0065 .*Passiflorasubpeltata (Kunth) L.H.Bailey \u2013 Life form: Herbaceous climber. Habitat: Moist forest edges, 1500\u20132060 m. Voucher: SK 0003 .PenaeaceaeF106. Oliniarochetiana A.Juss. \u2013 Life form: Tree. Habitat: Upland dry and moist evergreen forest, 1700\u20133100 m. Vouchers: Verdcourt 3283, Holyoak 712 (EA).PhytolaccaceaeF107. Phytolaccadodecandra L\u2019H\u00e9r. \u2013 Life form: Shrub. Habitat: Riparian vegetation and bushland, 500\u20132400 m. Vouchers: SK 0032, SK 0236 .PiperaceaeF108. Peperomiaabyssinica Miq. \u2013 Life form: Herb. Habitat: Moist montane forest, 1600\u20132950 m. Voucher: SK 0142 .Pipercapense L.f. \u2013 Life form: Herb or subshrub. Habitat: Upland swampy forest edges and wet forest floors, 1200\u20132700 m. Voucher: SK 0134 .PittosporaceaeF109. Pittosporumviridiflorum Sims \u2013 Life form: Tree. Habitat: Upland moist forest, 900\u20132400 m. Voucher: Faden et al. 74/858 (EA).PlantaginaceaeF110. Callitricheoreophila Schotsman \u2013 Life form: Herb. Habitat: Aquatic in water pools and streams, 1150\u20133300 m. Voucher: Chandler 2322 (EA).Callitrichevulcanicola Schotsman \u2013 Life form: Herb. Habitat: Moist grounds in montane grassland, 3000\u20134050 m. Vouchers: Hedberg 1650, Gilbert and Thulin 1047 (EA).Plantagopalmata Hook.f. \u2013 Life form: Herb. Habitat: Roadsides and clearings in upland forest, 1170\u20133300 m. Voucher: Bally 8519 (EA).Veronicaabyssinica Fresen. \u2013 Life form: Herb. Habitat: Upland forest and bushy grassland, 1200\u20133900 m. Vouchers: SK 0050, SAJIT 006464 .Veronicaanagallis-aquatica L. \u2013 Life form: Herb. Habitat: Stream-sides, 480\u20132400 m. Voucher: Glover & Samuel 445 (EA).Veronicaglandulosa Hochst. ex Benth. \u2013 Life form: Herb. Habitat: Upland forest margins, 2850\u20133980 m. Voucher: Mabberlay 350 (EA).PolygalaceaeF111. Polygalaohlendorfiana Eckl. & Zeyh. \u2013 Life form: Herb. Habitat: Upland grassland, 1800\u20133050 m. Voucher: Beentje 2658 (EA).Polygalasadebeckiana G\u00fcrke \u2013 Life form: Herb or subshrub. Habitat: Upland grassland, 10\u20132500 m. Voucher: SK 0257 .Polygalasphenoptera Fresen. \u2013 Life form: Herb or subshrub. Habitat: Wooded grassland and bushland, 0\u20133300 m. Voucher: Kokwaro 2797 (EA).Polygalasteudneri Chodat \u2013 Life form: Herb. Habitat: Upland grassland, 3000\u20134050 m. Voucher: Coe 775 (EA).PolygonaceaeF112. Harpagocarpussnowdenii Hutch. & Dandy \u2013 Life form: Herbaceous climber. Habitat: Upland moist forest, 1350\u20132650 m. Voucher: Gedye 6700 (EA).Oxygonumsinuatum (Hochst. & Steud ex Meisn.) Dammer \u2013 Life form: Herb. Habitat: Waste places in grassland, 0\u20132250 m. Voucher: Margareta 6 (EA).Oxygonumstuhlmannii Dammer \u2013 Life form: Herb. Habitat: Waste places in grassland, 10\u20132250 m. Voucher: Aement et al. 150 (EA).Persicariadecipiens (R.Br.) K.L.Wilson \u2013 Life form: Herb. Habitat: Wet places often in water pools, 1100\u20132291 m. Voucher: SK 0162 .Persicarianepalensis (Meisn.) H.Gross \u2013 Life form: Herb. Habitat: Upland grassland and forest edges, 1140\u20133500 m. Voucher: Kimani 9 (EA).Persicariasetosula (A.Rich.) K.L.Wilson \u2013 Life form: Herb. Habitat: Along streams in upland forest, 1050\u20132670 m. Voucher: Kerfoot 609 (EA).Persicariastrigosa (R.Br.) Nakai \u2013 Life form: Herb. Habitat: Upland moist forest and river banks, 1110\u20131920 m. Voucher: Faden 68/706 (EA).Polygonumafromontanum Greenway \u2013 Life form: Shrub. Habitat: Upland rainforest and moorland, 2100\u20133490 m. Voucher: Townsend 2316 (EA).Polygonumaviculare L. \u2013 Life form: Exotic herb. Habitat: Upland roadsides and disturbed areas, 2100\u20133490 m. Voucher: Albrechtsen 5238 (EA).*Rumexacetosella L. \u2013 Life form: Exotic herb. Habitat: Montane grassland, 2400\u20133160 m. Voucher: Luke 15363 (EA).*Rumexnepalensis Spreng. \u2013 Life form: Herb. Habitat: Upland grassland and bushland, 690\u20133700 m. Voucher: Blain 10915 (EA).Rumexruwenzoriensis Chiov. \u2013 Life form: Herb. Habitat: Upland grassland and moorland, 1950\u20133700 m. Voucher: SK 0172 .PortulacaceaeF113. Portulacanitida (Danin & H.G.Baker) Ricceri & Arrigoni \u2013 Life form: Herb. Habitat: Roadsides and waste places in grassland, 0\u20132350 m. Voucher: Faden 74/835 (EA).PrimulaceaeF114. Lysimachiaarvensis (L.) U.Manns & Anderb. \u2013 Life form: Herb. Habitat: Dry evergreen bushland, 1350\u20132635 m. Voucher: Verdcourt 715 (EA).Lysimachiahexamera (P.Taylor) U.Manns & Anderb. \u2013 Life form: Herb. Habitat: Swampy sites in upland grassland, 2100\u20132600 m. Voucher: Hedberg 1085(EA).Lysimachiaserpens (Hochst. ex A.DC.) U.Manns & Anderb. \u2013 Life form: Herb. Habitat: Damp sites in moorland, 2600\u20133960 m. Vouchers: Kuchar 9604 (EA), SK 0074 .Ardisiandrasibthorpioides Hook.f. \u2013 Life form: Herb. Habitat: Moist evergreen forest, 900\u20132670 m. Voucher: Chandler 2227 (EA).Ardisiandrawettsteinii J.Wagner \u2013 Life form: Herb. Habitat: Upland moist forest and bamboo thickets, 1580\u20133600 m. Voucher: Albrechtsen 5946 (EA).EEmbeliakeniensis R.E.Fr. \u2013 Life form: Tree. Habitat: Upland moist forest, 1500\u20132100 m. Voucher: Luke 447 (EA).Lysimachiaruhmeriana Vatke \u2013 Life form: Herb. Habitat: Wet montane forest, 2020\u20133500 m. Voucher: Kirika and York 1061 (EA).Maesalanceolata Forssk. \u2013 Life form: Shrub or tree. Habitat: Riverine forest and moist forest margins, 360\u20132800 m. Voucher: Wimbush 1118 (EA).Myrsineafricana L. \u2013 Life form: Shrub. Habitat: Upland wooded grassland, 1200\u20133600 m. Voucher: SK 0055 .Rapaneamelanophloeos (L.) Mez \u2013 Life form: Tree. Habitat: Moist forest, 900\u20133800 m. Voucher: SK 0101 .ProteaceaeF115. Faureaarborea Engl. \u2013 Life form: Tree. Habitat: Upland dry forest, 1280\u20133100 m. Voucher: Gardner 7063 (EA).Faurearochetiana (A.Rich.) Chiov. ex Pic.Serm. \u2013 Life form: Tree. Habitat: Wooded grassland, 900\u20132400 m. Voucher: Dyson 745 (EA).Faureasaligna Harv. \u2013 Life form: Tree. Habitat: Grassland with scattered trees, 700\u20131800 m. Voucher: Bono 10 (EA).Proteacaffrasubsp.kilimandscharica (Engl.) Chisumpa & Brummitt \u2013 Life form: Shrub. Habitat: Montane grassland and forest edges, 2300\u20133700 m. Vouchers: Beentje 3241 (EA), SK 0110 .Proteagaguedi J.F.Gmel. \u2013 Life form: Tree. Habitat: Woodland and scattered-tree grassland, 900\u20132100 m. Voucher: Holyoak 711 (EA).PutranjivaceaeF116. Drypetesgerrardiivar.gerrardii Hutch. \u2013 Life form: Tree. Habitat: Dry or moist evergreen forest, 1150\u20132300 m. Voucher: Fries 234 (EA).Drypetesgerrardiivar.tomentosa Radcl.-Sm. \u2013 Life form: Tree. Habitat: Upland evergreen forest, 1150\u20132000 m. Voucher: Kirika 496 (EA).RanunculaceaeF117. Anemonethomsonii Oliv. \u2013 Life form: Herb. Habitat: Wet rocky sites in moorland, 2500\u20134000 m. Vouchers: SK 0182, SK 0054 .Clematissimensis Fresen. \u2013 Life form: Woody climber. Habitat: Bushland and forest margins, 1000\u20133360 m. Vouchers: SK 0027, SK 0081 .Delphiniummacrocentrum Oliv. \u2013 Life form: Herb. Habitat: Upland grassland and bamboo thicket margins, 1650\u20133900 m. Voucher: SK 0107 .ERanunculusaberdaricus Ulbr. \u2013 Life form: Herb. Habitat: Clearings in moist bamboo forest, 2550\u20133660 m. Voucher: Coe 789 (EA).Ranunculusmultifidus Forssk. \u2013 Life form: Herb. Habitat: Stream banks and moist bushland, 1170\u20133450 m. Voucher: Mathenge 23 (EA).Ranunculusoreophytus Delile \u2013 Life form: Herb. Habitat: Wet and boggy places in moorland, 2240\u20134200 m. Voucher: Milne-Redhead et al. 1617 (EA).Ranunculusstagnalis Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Bogs and water pools in moorland, 3000\u20134750 m. Voucher: Hedberg 994 (EA).Ranunculusvolkensii Engl. \u2013 Life form: Herb. Habitat: Marshy sites in moorland, 2700\u20134050 m. Voucher: SK 0119 .Thalictrumrhynchocarpum Quart.-Dill. & A.Rich. \u2013 Life form: Herb. Habitat: Undergrowth in upland forest, 1550\u20133275 m. Voucher: SK 0145 .ResedaceaeF118. Cayluseaabyssinica (Fresen.) Fisch. & C.A.Mey. \u2013 Life form: Herb. Habitat: Roadsides in upland grassland, 1100\u20133000 m. Voucher: SK 0124 .RhamnaceaeF119. Helinusmystacinus (Aiton.) E.Mey. ex Steud. \u2013 Life form: Woody climber. Habitat: Wooded grassland and forest margins, 100\u20132400 m. Voucher: SK 0019 .Rhamnusprinoides L\u2019H\u00e9r. \u2013 Life form: Tree. Habitat: Upland moist forests and bushland, 1500\u20133700 m. Vouchers: SK 0006, SK 0263 .Rhamnusstaddo A.Rich. \u2013 Life form: Tree. Habitat: Upland evergreen bushland and dry forest margins, 1000\u20133600 m. Voucher: SAJIT 006504 .Scutiamyrtina (Burm.f.) Kurz \u2013 Life form: Shrub. Habitat: Forest margins and bushland, 0\u20132750 m. Voucher: SK 0266 .RhizophoraceaeF120. Cassipoureacelastroides Alston \u2013 Life form: Shrub or small tree. Habitat: Rocky hills in evergreen bushland, 250\u20131850 m. Voucher: Fries 2103 (EA).Cassipoureagummiflua Tul. \u2013 Life form: Tree. Habitat: Upland wet evergreen forest, 2000\u20132300 m. Vouchers: Luke et al. 7171, Medley 665 (EA).Cassipoureamalosana (Baker.) Alston \u2013 Life form: Tree. Habitat: Moist or dry forest, 750\u20132600 m. Voucher: SK 0025 .RosaceaeF121. Alchemillaargyrophylla Oliv. \u2013 Life form: Shrub. Habitat: Damp sites in moorland, 2250\u20134650 m. Vouchers: Vorontsova 43 (EA), SK 0053 .Alchemillacryptantha Steud. ex A.Rich. \u2013 Life form: Herb. Habitat: Moist moorland and bamboo thickets, 1300\u20134050 m. Voucher: Townsend 2436 (EA).Alchemillaabyssinicasubsp.cyclophylla (T.C.E.Fr.) Kalheber \u2013 Life form: Herb. Habitat: Moist moorland and bamboo thickets, 2900\u20134300 m. Voucher: SK 0095 .Alchemillaelgonensis Mildbr. \u2013 Life form: Shrub. Habitat: Moist moorland and bamboo thickets, 2700\u20134250 m. Voucher: Verdcourt 3777 (EA).Alchemillaellenbeckii Engl. \u2013 Life form: Herb. Habitat: Moist moorland and bamboo thickets, 2100\u20133900 m. Voucher: Hedberg 1501 (EA).Alchemillafischeri Engl. \u2013 Life form: Herb. Habitat: Moist montane forest and bamboo thickets, 2320\u20133440 m. Voucher: Muasya et al. 018(EA).Alchemillapedata Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Damp sites in upland grassland, 2120\u20133120 m. Voucher: Napier 683 (EA).Alchemillahageniae T.C.E.Fr. \u2013 Life form: Herb. Habitat: Moist bamboo thickets and montane evergreen bushland, 3000\u20133490 m. Voucher: Muasya et al. 052 (EA).Alchemillajohnstonii Oliv. \u2013 Life form: Shrub. Habitat: Moist moorland and bamboo thickets, 2400\u20134260 m. Voucher: Mabberley 324 (EA).Alchemillakiwuensis Engl. \u2013 Life form: Herb. Habitat: Moist upland forest and bamboo forest, 1250\u20133000 m. Voucher: Verdcourt 601 (EA).Alchemillamicrobetula T.C.E.Fr. \u2013 Life form: Herb. Habitat: Moist moorland, 3350\u20134400 m. Voucher: Hedberg 4430 (EA).Alchemillarothii Oliv. \u2013 Life form: Herb. Habitat: Moorland and upper edges of montane forest, 2700\u20134000 m. Voucher: Kuchar 12468(EA).Cliffortianitidula (Engl.) R.E.Fr. & T.C.E.Fr. \u2013 Life form: Shrub. Habitat: Damp sites in moorland, 2040\u20133150 m. Voucher: Kuchar 10289 (EA).Fragariavesca L. \u2013 Life form: Herb. Habitat: Upland grassland and forest edges, 2400\u20132850 m. Voucher: Mungai 50 (EA).Hageniaabyssinica (Bruce ex Steud.) J.F.Gmel. \u2013 Life form: Tree. Habitat: Upland moist forest, 2400\u20133600 m. Voucher: SK 0122 .Prunusafricana (Hook.f.) Kalkman \u2013 Life form: Tree. Habitat: Upland moist forest and riverine forest, 1350\u20132750 m. Voucher: Moon 765 (EA).Rosarubiginosa L. \u2013 Life form: Herb. Habitat: Upland forest margins, 1600\u20133000 m. Voucher: SK 0264 .Rubusapetalus Poir. \u2013 Life form: Shrub. Habitat: Upland moist forest, 1275\u20132700 m. Voucher: Logic/Bally 7958 (EA).Rubusfriesiorum Gust. \u2013 Life form: Shrub. Habitat: Upland moist forest, 3050\u20133400 m. Vouchers: Kuchar 12476 (EA), SK 0062 .Rubuskeniensis Standl. \u2013 Life form: Shrub. Habitat: Upland moist forest, 1950\u20132800 m. Voucher: SAJIT 006472 .Rubuspinnatus Willd. \u2013 Life form: Shrub. Habitat: Upland moist forest and bamboo thickets, 2400\u20133000 m. Voucher: Kuchar 7838 (EA).Rubusscheffleri Engl. \u2013 Life form: Shrub. Habitat: Upland moist forest margins and evergreen bushland, 1650\u20133150 m. Voucher: Grant 1217 (EA).Rubussteudneri Schweinf. \u2013 Life form: Shrub. Habitat: Upland moist forest margins and evergreen bushland, 1500\u20133480 m. Vouchers: Kuchar 8311, Kuchar and Msafiri 5206 (EA).Rubusvolkensii Engl. \u2013 Life form: Shrub. Habitat: Upland moist forest margins and open sites, 2100\u20133450 m. Voucher: SAJIT 006480 .RubiaceaeF122. Anthospermumherbaceum L.f. \u2013 Life form: Herb. Habitat: Woodland and forest edges, 900\u20133240 m. Voucher: Kerfoot 477 (EA).Anthospermumusambarense K.Schum. \u2013 Life form: Shrub. Habitat: Moorland and upper edges of montane forest, 1300\u20134050 m. Voucher: Gardner 1113 (EA).Canthiumoligocarpumsubsp.friesiorum (Robyns) Bridson \u2013 Life form: Shrub or tree. Habitat: Upland wet evergreen forest, 2000\u20132500 m. Vouchers: Battiscombe 1049, Venn Fey 11703 (EA).Chassaliakenyensis Verdc. \u2013 Life form: Shrub. Habitat: Moist evergreen forest, 1650\u20132300 m. Voucher: Polhill 360 (EA).Galinierasaxifraga (Hochst.) Bridson \u2013 Life form: Tree. Habitat: Upland moist forest and stream-sides, 1700\u20133000 m. Voucher: SK 0234 .Galiumacrophyum Hochst. ex Chiov. \u2013 Life form: Herb. Habitat: Open areas in bamboo thickets and montane forest, 2700\u20133200 m. Voucher: Kokwaro 1947 (EA).Galiumaparinoides Forssk. \u2013 Life form: Herb. Habitat: Wet evergreen forest, 1680\u20133700 m. Voucher: Verdcourt, Cooley and Howard 3766G (EA).Galiumglacialevar.satimmae Verdc. \u2013 Life form: Herb. Habitat: Damp places in moorland, 3510\u20134350 m. Voucher: Coe & Kirika 284 (EA).Galiumkenyanum Verdc. \u2013 Life form: Herb. Habitat: Upper margins of montane forest and moorland, 2880\u20133550 m. Voucher: Polhill 239 (EA).Galiumossirwaense K.Krause \u2013 Life form: Herb. Habitat: Upland moist forest edges, 2160\u20133750 m. Vouchers: Kuchar 12353, Napper 536 (EA).Galiumruwenzoriense (Cortesi) Chiov. \u2013 Life form: Herb. Habitat: Open upland forests or bushland, 2700\u20134000 m. Voucher: Hansen 870 (EA).Galiumsimense Fresen. \u2013 Life form: Herb. Habitat: Upland bushland and woodland, 1500\u20132700 m. Voucher: Taiti 2027 (EA).Galiumspuriumsubsp.africanum Verdc. \u2013 Life form: Herb. Habitat: Upland bushland and forest edges, 1250\u20132700 m. Vouchers: Knox 3213, Kanore Kibui 16 (EA).Galiumthunbergianum Eckl. & Zeyh. \u2013 Life form: Herb. Habitat: Wet montane forest, 2000\u20133750 m. Voucher: Knox 3091 (EA).Galiumscioanum Chiov. \u2013 Life form: Herb. Habitat: Upland swamps and riversides, 1800\u20132700 m. Voucher: SK 0093 .Lasianthuskilimandscharicus K.Schum. \u2013 Life form: Shrub or small tree. Habitat: Undergrowth in moist forest, 1500\u20132500 m. Voucher: Hanse 834 (EA).Mussaendamicrodontasubsp.odorata (Hutch.) Bridson \u2013 Life form: Shrub or tree. Habitat: Upland moist evergreen forest and riparian forest, 1830\u20132100 m. Vouchers: Greenway 9680, Kirika et al. 6 (EA).Oldenlandiafriesiorum Bremek \u2013 Life form: Herb. Habitat: Upland evergreen forest and forest edges, 1800\u20132550 m. Voucher: Kuchar 8348 (EA).Oldenlandiamonanthos (Hochst. ex A.Rich.) Hiern \u2013 Life form: Herb. Habitat: Upland evergreen forest and montane grassland, 1350\u20133500 m. Voucher: Lind 2883 (EA).Pauridianthapaucinervis (Hiern) Bremek. \u2013 Life form: Shrub or small tree. Habitat: Upland moist evergreen forest, 500\u20132400 m. Voucher: SK 0244 .Pavettaabyssinicavar.lamurensis Verdc. \u2013 Life form: Tree. Habitat: Upland forest and bushland, 1500\u20132550 m. Vouchers: Luke 705, Gardner 2837 (EA).Pentanisiafoetida Verdc. \u2013 Life form: Herb. Habitat: Upland grassland and forest margins, 1830\u20132300 m. Voucher: De Block & Stieperaere 503 (EA).Pentaslanceolata (Forssk.) Deflers \u2013 Life form: Herb or subshrub. Habitat: Roadsides in upland grassland, 1200\u20132830 m. Voucher: Young 1004 (EA).Psychotriafractinervata E.M.A.Petit \u2013 Life form: Tree. Habitat: Upland wet forest, 1800\u20132600 m. Vouchers: SK 0128, SK 0135, SK 0241 .Psychotriakirkii Hiern \u2013 Life form: Shrub. Habitat: Moist forests and open woodland, 250\u20132250 m. Voucher: SK 0152 .Psychotriamahonii C.H.Wright \u2013 Life form: Tree. Habitat: Upland wet evergreen forest and swampy forest, 1230\u20132700 m. Voucher: SK 0243 .Psydraxparviflorasubsp.rubrocostata (Robyns) Bridson \u2013 Life form: Tree. Habitat: Upland moist forest, 1375\u20132750 m. Vouchers: Someren 3534, Muhia 123 (EA).Psydraxschimperiana (A.Rich.) Bridson \u2013 Life form: Tree. Habitat: Moist forest and bushland, 15\u20132500 m. Voucher: SK 0005 .Rothmanniamanganjae (Hiern) Keay \u2013 Life form: Tree. Habitat: Moist forest, 230\u20131800 m. Voucher: Gardner 2477 (EA).Rubiacordifoliasubsp.conotricha (Gand.) Verdc. \u2013 Life form: Herbaceous climber. Habitat: Upland forest edges and bushland, 1140\u20133120 m. Vouchers: Verdcourt 620 (EA), SAJIT 006503 Rytigyniabugoyensis (K.Krause) Verdc. \u2013 Life form: Shrub. Habitat: Montane evergreen forest, 1230\u20132400 m. Voucher: Faden 67/142 (EA).Rytigyniauhligii (K.Schum. & K.Krause) Verdc. \u2013 Life form: Tree. Habitat: Montane dry or moist evergreen forest, 1000\u20132460 m. Voucher: SK 0200 .Spermacoceprinceae (K.Schum.) Verdc. \u2013 Life form: Herb. Habitat: Roadsides in bamboo thickets, 960\u20132650 m. Voucher: SK 0049 .Vangueriaapiculata K.Schum. \u2013 Life form: Tree. Habitat: Evergreen forest, bushland and riverine forest, 900\u20132330 m. Voucher: SK 0138 .Vangueriainfausta Burch. \u2013 Life form: Tree. Habitat: Evergreen forest margins and woodland, 30\u20132100 m. Voucher: Faden 6774 (EA).Vangueriamadagascariensis J.F.Gmel. \u2013 Life form: Tree. Habitat: Moist forest and riparian forest, 0\u20132130 m. Voucher: SK 0037 .Vangueriavolkensii K.Schum. \u2013 Life form: Tree. Habitat: Dry forest margin and riverine forest, 900\u20132300 m. Voucher: SK 0235 .RutaceaeF123. Calodendrumcapense (L.f.) Thunb. \u2013 Life form: Tree. Habitat: Upland evergreen and riverine forest, 1200\u20132200 m. Voucher: Kamau 326 (EA).Clausenaanisata (Willd.) Hook.f. ex Benth. \u2013 Life form: Tree. Habitat: Upland moist forest, 0\u20132700 m. Voucher: SK 0129 .Fagaropsisangolensis (Engl.) Dale \u2013 Life form: Tree. Habitat: Upland moist forest, 1000\u20132250 m. Voucher: Faden 74/895E (EA).Toddaliaasiatica (L.) Lam. \u2013 Life form: Shrub. Habitat: Moist forest edges and wet bushland, 0\u20133000 m. Voucher: SK 0002 .Veprisglandulosa (Hoyle & Leakey) Kokwaro \u2013 Life form: Tree. Habitat: Upland dry evergreen forest, 1700\u20132020 m. Voucher: Kirika 491 (EA).Veprishanangensisvar.unifoliata Kokwaro \u2013 Life form: Tree. Habitat: Upland evergreen forest and bushland, 1700\u20132020 m. Vouchers: Kokwaro 4038 & 4039, Greenway 7595 (EA).Veprisnobilis (Delile) Mziray \u2013 Life form: Tree. Habitat: Dry evergreen forest and bushland, 900\u20132750 m. Voucher: SK 0169 .Veprissimplicifolia (Engl.) Mziray \u2013 Life form: Tree. Habitat: Dry forest and evergreen forest and bushland, 300\u20132420 m. Voucher: Battiscombe 867 (EA).Vepristrichocarpa (Engl.) Mziray \u2013 Life form: Shrub. Habitat: Wooded grassland and riparian forest, 0\u20132660 m. Voucher: Stuhlmann 937 (EA).Zanthoxylumusambarense (Engl.) Kokwaro \u2013 Life form: Tree. Habitat: Upland dry forest, 1200\u20132600 m. Voucher: Greenway 12624 (EA).SalicaceaeF124. Caseariabattiscombei R.E.Fr. \u2013 Life form: Tree. Habitat: Upland moist forest, 1000\u20132440 m. Voucher: Gachathi 1/81 (EA).Dovyalisabyssinica (A.Rich.) Warb. \u2013 Life form: Shrub or tree. Habitat: Upland moist forest, 1450\u20133000 m. Voucher: SK 0021 .Oncobaroutledgei Sprague \u2013 Life form: Shrub or small tree. Habitat: Upland moist forest and riparian vegetation, 900\u20132440 m. Voucher: Hargen 1186 (EA).Oncobaspinosa Forssk. \u2013 Life form: Shrub. Habitat: Moist forest edges and bushland, 0\u20131800 m. Voucher: Kirika et al. 168 (EA).Scolopiazeyheri (Nees) Szyszy\u0142 \u2013 Life form: Tree. Habitat: Dry evergreen forest and bushland, 0\u20132400 m. Voucher: Verdcourt 3132 (EA).Trimeriagrandifolia (Hochst.) Warb. \u2013 Life form: Tree. Habitat: Dry evergreen forest and bushland, 150\u20132500 m. Voucher: Napier 1933 (EA).SantalaceaeF125. Osyrislanceolata Hochst. & Steud. \u2013 Life form: Shrub or small tree. Habitat: Upland forest and bushland, 900\u20132700 m. Voucher: Robertson 7675 (EA).Thesiumkilimandscharicum Engl. \u2013 Life form: Herb. Habitat: Montane grassland and moorland, 2200\u20134200 m. Voucher: Polhill 231 (EA).Thesiummukense A.W.Hill \u2013 Life form: Herb. Habitat: Upland grassland and woodland, 1100\u20132500 m. Voucher: Faden & Evans 74/622 (EA).Thesiumradicans Hochst. ex A.Rich. \u2013 Life form: Herb. Habitat: Upland open grassland, 1800\u20133000 m. Voucher: Harwey Albuchtsen 186 (EA).Thesiumschweinfurthii Engl. \u2013 Life form: Herb. Habitat: Upland grassland and woodland, 1050\u20132300 m. Voucher: Harwey Albuchtsen 187 (EA).Viscumtuberculatum A.Rich. \u2013 Life form: Shrub. Habitat: Upland dry evergreen forest and bushland, 1200\u20132400 m. Voucher: SK 0199 .SapindaceaeF126. Allophylusabyssinicus (Hochst.) Radk. \u2013 Life form: Tree. Habitat: Upland moist forest, 1050\u20132550 m. Voucher: Lind et al. 5029 (EA).Allophylusafricanus P.Beauv. \u2013 Life form: Tree. Habitat: Wooded grassland and forest margins, 650\u20132400 m. Voucher: SK 0194 .Blighiaunijugata Baker \u2013 Life form: Tree. Habitat: Moist or dry evergreen forest and bushland, 0\u20131900 m. Voucher: Battiscombe 6 (EA).Deinbolliakilimandscharica Taub. \u2013 Life form: Tree. Habitat: Moist or dry evergreen forest and bushland, 1100\u20132400 m. Voucher: E.A.A. F.RO Staff 53 (EA).Dodonaeaviscosasubsp.angustifolia (L.f.) J.G.West. \u2013 Life form: Shrub. Habitat: Grassland and bushland, 0\u20132700 m. Voucher: SK 0098 .SapotaceaeF127. Chrysophyllumgorungosanum Engl. \u2013 Life form: Tree. Habitat: Upland moist forest, 1300\u20132250 m. Voucher: Hockliffe 1370 (EA).Pouteriaadolfi-friedericiisubsp.keniensis (R.E.Fr.) L.Gaut. \u2013 Life form: Tree. Habitat: Upland moist forest, 1430\u20132500 m. Vouchers: Leaky 1224, Luke 408 (EA).ScrophulariaceaeF128. Hedbergiadecurva (Hochst. ex Benth.) A.Fleischm. & Heubl \u2013 Life form: Herb. Habitat: Moorland and upper edges of montane forest, 2950\u20133840 m. Voucher: SK 0078 .Hedbergialongiflora (Hochst. ex Benth.) A.Fleischm. & Heubl \u2013 Life form: Herb. Habitat: Moist montane forest and grassland, 2450\u20133500 m. Voucher: Kuchar 12763 (EA).Bartsiatrixago L. \u2013 Life form: Herb. Habitat: Rocky sites in moorland, 930\u20133700 m. Voucher: Dale 2683 (EA).Craterostigmapumilum Hochst. \u2013 Life form: Herb. Habitat: Montane grassland, 2000\u20132600 m. Voucher: Verdcourt 1042 (EA).Cycniumtenuisectum (Standl.) O.J.Hansen \u2013 Life form: Herb. Habitat: Open places in montane forest, 1800\u20133500 m. Voucher: Nattrass 1459 (EA).Cycniumtubulosumsubsp.montanum (N.E.Br.) O.J.Hansen. \u2013 Life form: Herb. Habitat: Grassland, 200\u20132600 m. Vouchers: Harvey 178, Tallantire 675 (EA).Diclisbambuseti R.E.Fr. \u2013 Life form: Herb. Habitat: Moist montane forest, 2000\u20133720 m. Voucher: Kuchar 12729 (EA).Diclisovata Benth. \u2013 Life form: Herb. Habitat: Upland moist grassland and montane forest, 1300\u20133440 m. Voucher: John Terry 178 (EA).Hebenstretiaangolensis Rolfe \u2013 Life form: Herb. Habitat: Dry montane grassland and rocky heathland, 1500\u20134000 m. Voucher: SAJIT 006476, SK 0091 .Limosellaafricana Gl\u00fcck \u2013 Life form: Herb. Habitat: Aquatic in upland waterfalls edges and stream banks, 1600\u20134200 m. Voucher: Gilbert et al. 1045 (EA).Limosellacapensis Thunb. \u2013 Life form: Herb. Habitat: Aquatic in muddy water pools and slow flowing streams, 1800\u20132800 m. Voucher: Verdcourt 641 (EA).Limosellamacrantha R.E.Fr. \u2013 Life form: Herb. Habitat: Moist depressions in montane forest and moorland, 2500\u20134500 m. Voucher: Napper 1232 (EA).Limosellamaior Diels \u2013 Life form: Herb. Habitat: Aquatic in upland muddy water pools, 1800\u20132700 m. Voucher: Greenway and Hemming 8768 (EA).Linderniarotundifolia (L.) Alston \u2013 Life form: Herb. Habitat: Roadsides in upland moist grassland, 1600\u20132000 m. Voucher: Agnew 7081 (EA).Selagothomsonii Rolfe \u2013 Life form: Herb. Habitat: Dry montane grassland, 1860\u20133380 m. Voucher: Greenway and Kanuri 15045 (EA).Sibthorpiaeuropaea L. \u2013 Life form: Herb. Habitat: Moist montane forest, 2000\u20133750 m. Voucher: SAJIT 006469 .Verbascumbrevipedicellatum (Engl.) Hub.-Mor. \u2013 Life form: Herb. Habitat: Upland rocky grassland, 1550\u20134100 m. Voucher: Mbale 860 (EA).Verbascumscrophulariifolium (Hochstetter) D.Hartl. \u2013 Life form: Herb. Habitat: Upland grassland, 1800\u20133600 m. Voucher: Piers 10163 (EA).SimaroubaceaeF129. Bruceaantidysenterica J.F.Mill. \u2013 Life form: Tree. Habitat: Upland dry or moist evergreen forest, 1400\u20132800 m. Voucher: Davidse 7054 (EA).SolanaceaeF130. Brugmansiasuaveolens (Humb. & Bonpl. ex Willd.) Sweet \u2013 Life form: Exotic shrub or small tree. Habitat: Forest margins and roadsides, 500\u20131800 m. Voucher: SK 0226 .*Cestrumaurantiacum Lindl. \u2013 Life form: Exotic shrub. Habitat: Roadsides and disturbed areas, 850\u20132600 m. Voucher: SK 0158 .*Cestrumelegans (Brongn.) Schltdl. \u2013 Life form: Exotic shrub or small tree. Habitat: Disturbed sites in upland moist forest, 1350\u20132600 m. Voucher: SK 0262 .*Discopodiumpenninervium Hochst. \u2013 Life form: Shrub. Habitat: Moist upper parts of montane forest and moorland, 1400\u20133000 m. Voucher: SK 0082 .Physalisphiladelphica Lam. \u2013 Life form: Exotic herb. Habitat: Forest margins and bushland, 990\u20132250 m. Voucher: SK 0031 .*Physalisperuviana L. \u2013 Life form: Exotic herb. Habitat: Upland forest margins and bushland, 1700\u20132500 m. Voucher: Kamau 333 (EA).*Solanumaculeastrum Dunal \u2013 Life form: Shrub or tree. Habitat: Upland forest margins and grassland, 1200\u20132500 m. Voucher: SK 0043 .Solanumagnewiorum Voronts. \u2013 Life form: Shrub. Habitat: Moist montane forest, 1800\u20132500 m. Voucher: Hepper and Field 4923 (EA).Solanumamericanum Mill. \u2013 Life form: Exotic herb. Habitat: Disturbed places in grassland and bushland, 0\u20133200 m. Voucher: Kerfoot 638 (EA).*Solanumgiganteum Jacq. \u2013 Life form: Shrub. Habitat: Upland grassland and bushland, 1500\u20132450 m. Voucher: Polhill & Verdcourt 267 (EA).Solanumincanum L. \u2013 Life form: Shrub. Habitat: Bushland and wooded grassland, 15\u20132200 m. Voucher: Bally and Smith 14628A (EA).Solanumlaxum Spreng. \u2013 Life form: Exotic shrub. Habitat: Upland rainforest, 1600\u20132650 m. Voucher: Verdcourt 3026 (EA).*Solanummauense Bitter \u2013 Life form: Shrub. Habitat: Upland forest edges and bushland, 1800\u20133000 m. Voucher: Simpson 13390 (EA).Solanummauritianum Scop. \u2013 Life form: Exotic shrub or small tree. Habitat: Upland forest margins and roadsides, 1150\u20132800 m. Voucher: SK 0151 .*Solanumphoxocarpum Voronts. \u2013 Life form: Shrub or tree. Habitat: Upland moist forest and woodland, 2100\u20133000 m. Voucher: Mbale et al. 844 (EA).Solanumpseudocapsicum L. \u2013 Life form: Exotic shrub. Habitat: Disturbed sites in upland forest, 1600\u20132150 m. Voucher: SK 0153 .*Solanumpseudospinosum C.H.Wright \u2013 Life form: Shrub. Habitat: Upland grassland and bushland, 2100\u20133600 m. Voucher: Snowden 564 (EA).Solanumrunsoriense C.H.Wright \u2013 Life form: Shrub. Habitat: Upper parts of montane forest, 2400\u20133200 m. Voucher: Polhill 250 (EA).Solanumschumannianum Dammer \u2013 Life form: Shrub. Habitat: Upland forest clearings, roadsides and bushland, 1300\u20133000 m. Voucher: SK 0029 .Solanumtarderemotum Bitter \u2013 Life form: Herb. Habitat: Moist forest margins and along streams, 550\u20132950 m. Voucher: Kerfoot 637 (EA).Solanumterminale Forsk. \u2013 Life form: Shrub. Habitat: Riverine forest and bushland, 400\u20133220 m. Voucher: SK 0040 .Withaniasomnifera (L.) Dunal \u2013 Life form: Herb or subshrub. Habitat: Grassland and bushland, 0\u20132800 m. Voucher: SK 0024 .StilbaceaeF131. Hallerialucida L. \u2013 Life form: Shrub. Habitat: Dry forest, 900\u20132700 m. Voucher: SK 0104 .ThymelaeaceaeF132. Gnidiaglauca (Fresen.) Gilg \u2013 Life form: Tree. Habitat: Upland forest margins to bamboo zone, 2250\u20133300 m. Voucher: Kerfoot 1402 (EA).Peddieafischeri Engl. \u2013 Life form: Tree. Habitat: Upland forest understorey and bushland, 950\u20132400 m. Voucher: Battiscombe 391 (EA).UrticaceaeF133. Didymodoxacaffra (Thunb.) Friis & Wilmot-Dear \u2013 Life form: Herb. Habitat: Dry upland forests, 1750\u20132840 m. Voucher: Faden & Evans 74/682 (EA).Droguetiadebilis Rendle \u2013 Life form: Herb. Habitat: Undergrowth in upland moist forest, 1550\u20133090 m. Voucher: Agnew et al. 8181 (EA).Droguetiainers (Forssk.) Schweinf. \u2013 Life form: Herb. Habitat: Upland moist evergreen forest and edges of bamboo thicket, 1600\u20133250 m. Voucher: Agnew et al. 7118 (EA).Elatostemamonticola Hook.f. \u2013 Life form: Herb. Habitat: Upland moist evergreen forest, 1600\u20132800 m. Voucher: Kerfoot 601 (EA).Girardiniabullosa (Hochst. ex Steud.) Wedd. \u2013 Life form: Herb. Habitat: Upland moist forest and forest margins, 1800\u20132600 m. Voucher: Mathenge 212 (EA).Girardiniadiversifolia (Link) Friis \u2013 Life form: Herb. Habitat: Open sites in upland moist forest and margins, 1100\u20132500 m. Voucher: Agnew 7692 (EA).Laporteaalatipes Hook.f. \u2013 Life form: Herb. Habitat: Undergrowth in upland moist forest, 1400\u20133500 m. Voucher: Verdcourt 2307 (EA).Myrianthusholstii Engl. \u2013 Life form: Tree. Habitat: Upland moist forest, 900\u20132100 m. Voucher: Hutchins (EA).Obetiaradula (Baker) Baker ex B.D.Jacks. \u2013 Life form: Tree. Habitat: Dry forest margins, 700\u20132000 m. Voucher: Hansen 745 (EA).Parietariadebilis G.Forst. \u2013 Life form: Herb. Habitat: Upland moist forest and bamboo thickets, 1700\u20134200 m. Voucher: Kerfoot 655 (EA).Pilearivularis Wedd. \u2013 Life form: Herb. Habitat: Upland moist forest, 1250\u20133100 m. Voucher: Kerfoot 468 (EA).Pileausambarensisvar.veronicifolia (Engl.) Friis \u2013 Life form: Herb. Habitat: Upland moist forest, 2030\u20133160 m. Voucher: Agnew 7164 (EA).Pileausambarensisvar.engleri (Rendle) Friis \u2013 Life form: Herb. Habitat: Upland rainforest, 1500\u20133160 m. Voucher: Fries and Fries 2785 (EA).Pileajohnstonii Oliv. \u2013 Life form: Herb. Habitat: Moist montane forest, 1450\u20132910 m. Voucher: SAJIT 006462 (EA).Urticamassaica Mildbr. \u2013 Life form: Herb. Habitat: Moist montane forest, 400\u20133320 m. Voucher: SK 0038 .Urticaurens L. \u2013 Life form: Herb. Habitat: Disturbed sites in upland moist forest, 1800\u20132500 m. Voucher: Gillet 16765 (EA).VerbenaceaeF134. Glandulariaaristigera (S.Moore) Tronc. \u2013 Life form: Exotic herb. Habitat: Roadsides in upland forest, 1650\u20132060 m. Voucher: Mungai 106 (EA).*Lantanacamara L. \u2013 Life form: Exotic shrub. Habitat: Open areas or clearings in forest and roadsides, 0\u20132040 m. Voucher: SK 0201 .*Lantanatrifolia L. \u2013 Life form: Exotic shrub. Habitat: Grassland and bushland, 0\u20132400 m. Voucher: SK 0012 .*Lantanaviburnoidessubsp.viburnoides (Forssk.) Vahl \u2013 Life form: Woody herb or shrub. Habitat: Bushland, 0\u20131950 m. Vouchers: Kamau 76 (EA), SK 0011 .Lippiakituiensis Vatke \u2013 Life form: Woody herb or shrub. Habitat: Woodland and bushland, 405\u20132550 m. Voucher: Trapnell 2134 (EA).Verbenabrasiliensis Vell. \u2013 Life form: Herb. Habitat: Woodland and bushland, 1050\u20132220 m. Voucher: Gillet 16285A (EA).ViolaceaeF135. Violaabyssinica Steud. ex Oliv. \u2013 Life form: Herb. Habitat: Upland evergreen forest and bamboo thickets, 1200\u20133740 m. Voucher: Napier 723 (EA).Violaeminii (Engl.) R.E.Fr. \u2013 Life form: Herb. Habitat: Montane forest and moorlands, 2150\u20134050 m. Voucher: Napier 676 (EA).Violanannae R.E.Fr. \u2013 Life form: Herb. Habitat: Montane grassland and heathland, 2550\u20133620 m. Voucher: SAJIT 006482 .VitaceaeF136. Cyphostemmakilimandscharicum (Gilg) Desc. ex Wild & R.B.Drumm. \u2013 Life form: Herbaceous climber. Habitat: Moist montane forest and bamboo thickets, 1590\u20133040 m. Vouchers: SK 0045, SK 0076 .Cyphostemmamaranguense (Gilg) Desc. \u2013 Life form: Herbaceous climber. Habitat: Dry upland forest and scattered-tree grasslands, 1500\u20132300 m. Voucher: Kirrika 498 (EA)."} +{"text": "Biophys J. (2012) 102:2605\u201314. doi: 10.1016/j.bpj.2012.04.029.\u201d It should be \u201cBasu S, Bhattacharyya D, Banerjee R. Self-complementarity within proteins: bridging the gap between binding and folding. Biophys J. (2012) 102:2605\u201314. doi: 10.1016/j.bpj.2012.04.029.\u201dIn the original article, the reference for \u201c77\u201d was incorrectly written as \u201cBatu S, Bhattacharyya D, Banerjee R. Self-complementarity within proteins: bridging the gap between binding and folding. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "BMJ 2020;369:m1328) has been updated. For the latest update, visit doi:10.1136/bmj.m1328.This living systematic review by Wynants and colleagues ("} +{"text": "This article has been corrected: During the assembly of 45976-45994. https://doi.org/10.18632/oncotarget.10275Original article: Oncotarget. 2016; 7:45976\u201345994."} +{"text": "This article has been corrected: During assembly of 7930-7943. https://doi.org/10.18632/oncotarget.3181Original article: Oncotarget. 2015; 6:7930\u20137943."} +{"text": "JCI Insight. 2019;4(8):e126749. https://doi.org/10.1172/jci.insight.126749Original citation: JCI Insight. 2020;5(15):e142898. https://doi.org/10.1172/jci.insight.142898Citation for this erratum: When the article was originally published, an incorrect panel was included in JCI Insight regrets the error."} +{"text": "Mol Immunol. (2019) 105:116\u201330. doi: 10.1016/j.molimm.2018.09.023.\u201d It should be \u201cShissler SC, Lee MS, Webb TJ. Mixed signals: co-stimulation in invariant natural killer T cell-mediated cancer immunotherapy. Front Immunol. (2017) 8:1447. doi: 10.3389/fimmu.2017.01447\u201d.Due to a production error, reference 5 was incorrectly written as \u201cShissler SC, Webb TJ. The ins and outs of type I iNKT cell development. The publisher apologizes for this mistake. The original article has been updated."} +{"text": "Author E. Gabriele Panarelli\u2019s ORCID iD is: 0000-0001-8790-5328 (The fourth author's name is spelled incorrectly. The correct name is: Edgar J. J. Groenen. The publisher apologizes for the error."} +{"text": "This article has been corrected: Due to errors during image assembly, the images for 111144-111160. https://doi.org/10.18632/oncotarget.22676Original article: Oncotarget. 2017; 8:111144\u2013111160."} +{"text": "This article has been corrected: In 48050-48058. https://doi.org/10.18632/oncotarget.10347Original article: Oncotarget. 2016; 7:48050\u201348058."} +{"text": "This article has been corrected: Due to a naming mistake during the original image capture process, the 24-hour 20 microMolar image in 1603-1617. https://doi.org/10.18632/oncotarget.27558Original article: Oncotarget. 2020; 11:1603\u20131617."} +{"text": "In \u201cYouth Experiences With Referrals to Mental Health Services in Canada: Protocol for a Web-Based Cross-Sectional Survey Study\u201d :e16945) the authors noted a footnote was erroneously omitted from the text of https://www12.statcan.gc.ca/census-recensement/2016/ref/dict/app-ann/a5_1-eng.cfm, 2) Archived Census 2A-L - 2016. Retrieved from: http://www23.statcan.gc.ca/imdb/p3Instr.pl?Function=getInstrumentList& Item_Id=295122&UL=1V&, 3) Ethnic Origin Reference Guide, Census of Population, 2016. Retrieved from: https://www12.statcan.gc.ca/census-recensement/2016/ref/guides/008/98-500-x2016008-eng.cfm & 4) Data Tables 2016 Census: Ethnic Origins. Retrieved from: https://www12.statcan.gc.ca/census-recensement/2016/dp-pd/dt-td/Av-eng.cfm?LANG=E& APATH=3&DETAIL=0&DIM=1&FL=A&FREE=0& GC=0&GID=0&GK=0&GRP=1&PID=112450& PRID=10&PTYPE=109445&S=0& SHOWALL=0&SUB=0&Temporal=2017& THEME=120&VID=29591&VNAMEE=&VNAMEF=Ethnicity question and responses were adapted using information from the following documents: 1) Statistics Canada Census of Population 2016, Appendix 5.1 Ethnic Origins disseminated from 2016, 2011, and 2006. Retrieved from: The corrected"} +{"text": "Open Biol.10, 200234 (Published Online 28 October 2020) (doi:10.1098/rsob.200234)"} +{"text": "Correction to: J Occup Med Toxicolhttp://dx.doi.org/10.1186/s12995-017-0177-2After publication of our article we have Original name spelling:Guadio F.Correct name spelling:Gaudio F."} +{"text": "This article has been corrected: During the assembly of 2515-2526. https://doi.org/10.18632/oncotarget.23507Original article: Oncotarget. 2018; 9:2515\u20132526."} +{"text": "MeFT1 produces early flowering in cassava. PLoS ONE 15(1): e0227199. https://doi.org/10.1371/journal.pone.0227199.The second author\u2019s name in incorrect. The correct name is: Getu Beyene. The correct citation is: Odipio J, Beyene G, Chauhan RD, Alicai T, Bart R, et al. (2020) Transgenic overexpression of endogenous FLOWERING LOCUS T-like gene The publisher apologizes for the error."} +{"text": "This article has been corrected: Due to errors during figure assembly, the images for 79605-79616. https://doi.org/10.18632/oncotarget.12869Original article: Oncotarget. 2016; 7:79605\u201379616."} +{"text": "Nature Microbiology 10.1038/s41564-019-0510-x, published online 22 July 2019.Correction to: https://genome.jgi.doe.gov/portal/PhyloTag/PhyloTag.home.html\u2019 was incorrect; it should have been \u2018https://genome.jgi.doe.gov/portal/Inovirus/Inovirus.home.html\u2019. This has now been corrected.In the version of this Article originally published, in the Data availability section, the link \u2018"} +{"text": "This article has been corrected: The name of the 4th author in the listing has been updated, correcting the name as follows:4,5Antonio Herrera-Merchan82294-82302. https://doi.org/10.18632/oncotarget.19393Original article: Oncotarget. 2017; 8:82294\u201382302."} +{"text": "This article has been corrected: In 81285-81294. https://doi.org/10.18632/oncotarget.18150Original article: Oncotarget. 2017; 8:81285\u201381294."} +{"text": "Keynote at the 20th European Conference on Eye Movement Research (ECEM) in Alicante, 20.8.2019.Video stream:https://vimeo.com/357889739"} +{"text": "CLINICS 2020;75:e1759errhttps://doi.org/10.6061/clinics/2020/e1759, published in 2020.Erratum for: doi: Replace the corresponding author: Yanhua ShenFor: Haixing Jiang*shenyanhua99@163.comReplace the corresponding email: jihaxi@163.comFor:"} +{"text": "Correction to: EJNMMI Phys (2020) 7:24https://doi.org/10.1186/s40658-020-00295-xFollowing publication of the original article , the autThe incorrect author names are: C. G. McGinnity and S. M. GouldThe correct author names are: C. J. McGinnity and S-M GouldThe author group has been updated above and the original article has been"} +{"text": "This article has been corrected: The name of the 2nd author has been updated as follows:Rob W.J. Collin2930-2946. https://doi.org/10.18632/oncotarget.26873Original article: Oncotarget. 2019; 10:2930\u20132946."} +{"text": "This article has been corrected: In 74494-74505. https://doi.org/10.18632/oncotarget.20170Original article: Oncotarget. 2017; 8:74494\u201374505."} +{"text": "Biology Methods and Protocols, 2017, 2(1), bpx006. doi:10.1093/biomethods/bpx006http://ssb22.user.srcf.net/pooler/.The server people.ds.cam.ac.uk has been permanently shut down. The source codes and executables are now available at: The article has been updated to reflect this."} +{"text": "This article has been corrected: The co-first author, Cheng-Jei Lin, has been removed as an equal contributor for this study as compared to the first author.74320-74330. https://doi.org/10.18632/oncotarget.20382Original article: Oncotarget. 2017; 8:74320\u201374330."} +{"text": "Giardia isolates genetically among patients in Chandigarh region, India. For this, nested PCR targeting fragment of the glutamate dehydrogenase (GLUD1 earlier named as GDH) gene was used. Phylogenetic analysis was done by constructing neighbor-joining tree made out of the nucleotide sequences of G. intestinalis isolates obtained in this study and with the known sequences published in GenBank.The aim of study was to characterize GLUD1 gene was amplified in 33 samples (82.5%). The product of GLUD1 gene was successfully sequenced only in 32 samples. In these samples, assemblage B was found in 27 (84.37%) samples whereas 5 (15.6%) samples had assemblage A. Among assemblage B most of them were of BIII. Therefore, genotyping of Giardia would be helpful in conducting epidemiological studies.Out of 40 samples, Giardia intestinalis\u00a0is well known intestinal parasite of humans and mammals. Giardia\u00a0causes approximately 280 million cases of giardiasis\u00a0worldwide annually [annually . Most ofGiardia in water or food [Giardia intestinalis\u00a0is\u00a0composed of eight major genotypes or assemblages (A\u2013H) [GLUD1 (earlier known as GDH)\u00a0locus has been utilized for genetic characterization of G. intestinalis isolates in vertebrates [Giardia isolates involved in its transmission by using glutamate dehydrogenase (GLUD1) marker.Giardiasis is acquired due to ingestion of cysts of or food . Giardiaes (A\u2013H) . Genotypes (A\u2013H) . These aes (A\u2013H) . Assembles (A\u2013H) , 5. The tebrates hosts anGiardia positive stool samples were collected from the Routine Laboratory of Department of Medical Parasitology, PGIMER, Chandigarh from August 2019 to December 2019.Forty microscopic From stool samples, DNA was extracted by using QIAmp Fast DNA Stool Mini Kit as per manufacturer\u2019s instructions with slight modifications. The suspension was initially incubated at 90\u00a0\u00b0C for 15\u00a0min and then for another 30\u00a0min at 75\u00a0\u00b0C. DNA was eluted in 50\u00a0\u00b5l of AE buffer. DNA concentration was measured by NanoQuant (Infinite\u00ae 200 PRO NanoQuant) and stored at \u2212\u00a020 \u00b0C until\u00a0further use.GLUD1 gene (432\u00a0bp) by using previously published primers given by Read et al. [Giardia\u00a0strain, Portland 1 was used for each PCR reaction. All the precautions were taken to prevent contamination.The two-step PCR was employed for the amplification\u00a0of\u00a0 d et al. . The conhttp://www.ncbi.nlm.nih.gov/blast). CLUSTAL X was used to determine multiple sequence alignments. Neighbor-joining distance trees were prepared using MEGAX software (https://www.megasoftware.net/) Fig. . BootstrThe direct links which are publicly available are as follows:https://www.ncbi.nlm.nih.gov/nuccore/MT584168https://www.ncbi.nlm.nih.gov/nuccore/MT584169https://www.ncbi.nlm.nih.gov/nuccore/MT584170https://www.ncbi.nlm.nih.gov/nuccore/MT584171https://www.ncbi.nlm.nih.gov/nuccore/MT584172https://www.ncbi.nlm.nih.gov/nuccore/MT584173https://www.ncbi.nlm.nih.gov/nuccore/MT584174https://www.ncbi.nlm.nih.gov/nuccore/MT584175https://www.ncbi.nlm.nih.gov/nuccore/MT584176https://www.ncbi.nlm.nih.gov/nuccore/MT584177https://www.ncbi.nlm.nih.gov/nuccore/MT584178https://www.ncbi.nlm.nih.gov/nuccore/MT584179https://www.ncbi.nlm.nih.gov/nuccore/MT584180https://www.ncbi.nlm.nih.gov/nuccore/MT584181https://www.ncbi.nlm.nih.gov/nuccore/MT584182https://www.ncbi.nlm.nih.gov/nuccore/MT584183https://www.ncbi.nlm.nih.gov/nuccore/MT584184https://www.ncbi.nlm.nih.gov/nuccore/MT584185https://www.ncbi.nlm.nih.gov/nuccore/MT584186https://www.ncbi.nlm.nih.gov/nuccore/MT584187https://www.ncbi.nlm.nih.gov/nuccore/MT584188https://www.ncbi.nlm.nih.gov/nuccore/MT584189https://www.ncbi.nlm.nih.gov/nuccore/MT584190https://www.ncbi.nlm.nih.gov/nuccore/MT584191https://www.ncbi.nlm.nih.gov/nuccore/MT584192https://www.ncbi.nlm.nih.gov/nuccore/MT584193https://www.ncbi.nlm.nih.gov/nuccore/MT584194https://www.ncbi.nlm.nih.gov/nuccore/MT584195https://www.ncbi.nlm.nih.gov/nuccore/MT584196https://www.ncbi.nlm.nih.gov/nuccore/MT584197https://www.ncbi.nlm.nih.gov/nuccore/MT584198https://www.ncbi.nlm.nih.gov/nuccore/MT584199GLUD1 gene was amplified in 33 samples (82.5%) among which 18 (66.66%) were sub-genotype BIII and 9 (33.33%) were sub-genotype BIV samples whereas 5 (15.6%) samples had assemblage A. Four of them were AI subgenotype and only1 belonged to sub-genotype AII.Out of 40 samples, the samples 8.5% and potable water\u00a0resources of Northern India [.ountries . The pre studies , 9. In o studies . There arn India , 14. Butrn India . Due to Giardia intestinalis. Detection of Giardia intestinalis assemblages and sub-assemblageswould be helpful in conducting epidemiological studies.The results\u00a0showed that PCR sequencing and phylogenetic analysis is an excellent molecular technique for genotyping of Giardia isolates.Present study involves only single locus for genotyping and also the sample size is less so it is difficult to interpret zoonotic potential of these isolates. Therefore, multi-locus typing data is required to differentiate between Additional file 1: Table S1. Sequence of primers and sgRNA were listed."} +{"text": "This article has an addendum: No external funding was obtained.1109-1119. https://doi.org/10.18632/oncotarget.28280Original article: Oncotarget. 2022; 13:1109\u20131119."} +{"text": "The first author\u2019s name is spelled incorrectly. The correct name is: Zeeshan Anjum Memon.https://doi.org/10.1371/journal.pone.0264958The correct citation is: Memon ZA, Said DM, Hassan MY, Leghari ZH, Sahar G. (2022) Parallel operated hybrid Arithmetic-Salp swarm optimizer for optimal allocation of multiple distributed generation units in distribution networks. PloS ONE 17(4): e0264958."} +{"text": "Bursaphelenchus zvyagintsevi sp. n. The corrected main text appears below in Section 2.1:In the original publication , there wBursaphelenchus zvyagintsevi sp. n.Adults : http://zoobank.org/urn:lsid:zoobank.org:act:71BDE14D-B778-40FE-977D-D910F701DE6D (accessed on 13 August 2023). Body curved ventrally. Stylet base slightly expanded, but without distinct knobs. Cephalic annuli faintly distinct through light microscopy. Median bulb ellipsoid, large; valve median to sub-median of bulb. Excretory pore located at nerve ring or at posterior end of the median bulb. Lateral field with two incisures.The authors state that the scientific conclusions are unaffected. This correction was approved by the Academic Editor. The original publication has also been updated."} +{"text": "This article has been corrected: In 86075-86086. https://doi.org/10.18632/oncotarget.13342Original article: Oncotarget. 2016; 7:86075\u201386086."} +{"text": "This article has been corrected: In 858-874. https://doi.org/10.18632/oncotarget.27495Original article: Oncotarget. 2020; 11:858\u2013874."} +{"text": "This article has been corrected: In 11641-11658. https://doi.org/10.18632/oncotarget.14264Original article: Oncotarget. 2017; 8:11641\u201311658."} +{"text": "Correction: BMC Women\u2019s Health (2022) 22:110.1186/s12905-022-01750-wFollowing publication of the original article , the refEinenkel R, Zygmunt M, Muzzio DO. Microorganisms in the healthy upper reproductive tract: from denial to benefcial assignments for reproductive biology. Reprod Biol 2019;19(2):113\u2013118. 10.1016/j.repbio.2019.04.001. Epub 2019 Apr 22. PMID: 31023521.The original article has been corrected."} +{"text": "This article has been retracted: In Figure 8, panel B, three of the tumor images are duplicates of images published in three other journals [journals \u20133. In ad80757-80769. https://doi.org/10.18632/oncotarget.20358Original article: Oncotarget. 2017; 8:80757\u201380769."} +{"text": "Nature Communications 10.1038/s41467-023-40061-y, published online 24 July 2023Correction to: In this article the author name R. C. Coombes was incorrectly written as R. Charles Coombes. The original article has been corrected."} +{"text": "This article has been retracted: Several images in this paper are duplicates of images in other published papers. In Figure 5C, a panel is duplicated within Figure 6D of another paper [er paper . In Figuer paper . In Figuer paper . Althoug65823-65835. https://doi.org/10.18632/oncotarget.19502Original article: Oncotarget. 2017; 8:65823\u201365835."} +{"text": "Correction: Stem Cell Research & Therapy (2021) 12:62 10.1186/s13287-020-02104-9The original article mistakenFundingThis study was supported by FCT \u2013 Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia, I.P., component OE under the scope of project UIDB/50026/2020 and by FCT/MCTES through the PD/59/2013, PD/BD/113800/2015 (C.M. Abreu), CEECIND/00695/2017 (M.T. Cerqueira), IF/00347/2015 (R. P. Pirraco), and IF/00945/2014 (A.P. Marques) grants."} +{"text": "This article has an addendum: Wet-lab experiments conducted at Data Driven Bioscience, Rincon Biosciences, Reprocell Inc., intoDNA, and Toxys Europe were funded by Lantern Pharma Inc.597-611. https://doi.org/10.18632/oncotarget.28454Original article: Oncotarget. 2023; 14:597\u2013611."} +{"text": "Can fresh embryo transfers be replaced by cryopreserved-thawed embryo transfers in assisted reproductive cycles? A randomized controlled trial. J. Assist. Reprod. Genet. 27 (7), 357\u2013363. doi:10.1007/s10815-010-9412-9\u201d.In the published article, the It should be \u201cPereira N, Rosenwaks Z. (2016). A fresh(er) perspective on frozen embryo transfers. Fertil Steril. 106(2), 257\u2013258. doi: 10.1016/j.fertnstert.2016.06.028\u201d.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Correction to: Pediatric Radiology (2023)https://doi.org/10.1007/s00247-023-05618-5The initial online version states that all authors participated in the roundtable discussion.Correction:Only the following authors participated in the roundtable discussion.Chair: A.C.O.Panel/speakers: A.M.J., K.M., I.M., M.R.The original article has been corrected."} +{"text": "This article has an addendum: Informed patient consent was obtained.6038-6042. https://doi.org/10.18632/oncotarget.27203Original article: Oncotarget. 2019; 10:6038\u20136042."} +{"text": "Orgiani are co-correspondingauthors.F. Mazzola("} +{"text": "This article has been corrected: In 103261-103273. https://doi.org/10.18632/oncotarget.21143Original article: Oncotarget. 2017; 8:103261\u2013103273."} +{"text": "This article has been corrected: In 21663-21673. https://doi.org/10.18632/oncotarget.15523Original article: Oncotarget. 2017; 8:21663\u201321673."} +{"text": "The seasonal influenza activity has been dramatically reduced worldwide due to COVID\u201019 control measures.Viral culture and characterization were routinely performed as described previously.Between January and December 2022, a total of 60 influenza viruses were isolated and characterized Table\u00a0. Of the Influenza activity in Hong Kong remained to be low in 2022. Various COVID\u201019 control measures such as the universal masking policy were still in force at the time of writing this report.Gannon C.K. Mak: Conceptualization; data curation; formal analysis; investigation; methodology; writing\u2014original draft; writing\u2014review and editing. Stephen S.Y. Lau: Investigation. Kitty K.Y. Wong: Investigation. Angela W.L. Lau: Investigation. Derek L.L. Hung: Writing\u2014original draft; writing\u2014review and editing.None.https://www.webofscience.com/api/gateway/wos/peer-review/10.1111/irv.13123.The peer review history for this article is available at"} +{"text": "Scientific Reports 10.1038/s41598-023-34932-z, published online 24 May 2023Correction to: The Data availability section in the original version of this Article was incorrect. It now reads:\u201cEach of the used datasets can be found on their respective links as shown below:https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0231304 should be contacted.TFED: For the dataset, the authors of the original paper introducing the public version of the dataset at this address http://www.psy.ntu.edu.tw/vnl/paper/Chen_2009_Exodatabase_SPIE.pdf) should be contacted.TFEID: For the dataset, the authors of the original paper introducing the public version of the dataset (at this link https://zenodo.org/record/3451524JAFFE: https://www.kaggle.com/datasets/shawon10/ckplusCK+: https://www.kaggle.com/datasets/msambare/fer2013FER2013: https://www.oulu.fi/en/university/faculties-and-units/faculty-information-technology-and-electrical-engineering/center-machine-vision-and-signal-analysis\u201d.SMIC: The original Article has been corrected."} +{"text": "This article has an addendum: Informed patient consent to an IRB-approved research protocol was obtained.245-251. https://doi.org/10.18632/oncotarget.26521Original article: Oncotarget. 2019; 10:245\u2013251."} +{"text": "This article has been retracted: A number of figure images in this paper were previously published in earlier papers. Figure 2E contains images from Figure 6C of another paper [er paper . Figure er paper . Figure er paper . Images er paper . Figure er paper . In addi101649-101658. https://doi.org/10.18632/oncotarget.21417Original article: Oncotarget. 2017; 8:101649\u2013101658."} +{"text": "PLoS Genet 19(10): e1011011. https://doi.org/10.1371/journal.pgen.1011011The first author\u2019s name is spelled incorrectly. The correct name is: Lin Zhang. The correct citation is: Zhang L, Green EW, Webster SG, Hastings MH, Wilcockson DC, Kyriacou CP (2023) The circadian clock gene"} +{"text": "Please remember to self\u2010claim your CPD and retain your supporting evidence. Answers are available online at J Med Radiat Sci. 2022 Dec; 69(4): 421\u2013430. https://doi.org/10.1002/jmrs.607Castillo C, Steffens T, Livesay G, et\u00a0al. IMPACT requests: Delphi study to develop criteria standards for adequate clinical information in computed tomography requests in the Australian emergency department. https://doi.org/10.1002/jmrs.621CPD Questions J Med Radiat Sci. 2022 Dec; 69(4): 463\u2013472. https://doi.org/10.1002/jmrs.606Smith\u2010Lickess SK, Stefanic N, Shaw J, et\u00a0al. What is the effect of a low literacy talking book on patient knowledge, anxiety and communication before radiation therapy starts? A pilot study. https://doi.org/10.1002/jmrs.628CPD Questions J Med Radiat Sci. 2023 Mar; 70(1): 46\u201355. https://doi.org/10.1002/jmrs.623Pinson JA, King OA, Dennett AM, et\u00a0al. Exploring the role of medical imaging assistants in Australian medical imaging departments: a mixed\u2010methods study. https://doi.org/10.1002/jmrs.644CPD Questions J Med Radiat Sci. 2023 Mar; 70(1): 72\u201380. https://doi.org/10.1002/jmrs.634Mark F, Alnsour A, Penfold SN, et\u00a0al. Volumetric modulated arc therapy (VMAT) comparison to 3D\u2010conformal technique in lung stereotactic ablative radiotherapy (SABR). https://doi.org/10.1002/jmrs.659CPD Questions J Med Radiat Sci. 2023 Apr; 70(Suppl 2): 48\u201358. https://doi.org/10.1002/jmrs.617Phonlakrai M, Ramadan S, Simpson J, et\u00a0al. Determination of hepatic extraction fraction with gadoxetate low\u2010temporal resolution DCE\u2010MRI\u2010based deconvolution analysis: validation with ALBI score and Child\u2010Pugh class. https://doi.org/10.1002/jmrs.645CPD Questions J Med Radiat Sci. 2023 Apr; 70(Suppl 2): 15\u201325. https://doi.org/10.1002/jmrs.618Gibbons E, Hoffmann M, Westhuyzen J, et\u00a0al. Clinical evaluation of deep learning and atlas\u2010based auto\u2010segmentation for critical organs at risk in radiation therapy. https://doi.org/10.1002/jmrs.655CPD Questions J Med Radiat Sci. 2023 Jun; 70(2): 137\u2013144. doi: 10.1002/jmrs.654Bantas G, Sweeney RJ, Mdletshe S. Digital radiography reject analysis: a comparison between two radiology departments in New Zealand. doi: 10.1002/jmrs.679CPD Questions https://doi.org/10.1002/jmrs.652Laing B, Caldwell P, Vincent D, Rattray G. An evaluation of radiation therapy patient body mass index trends and potential impact on departmental resource planning. J Med Radiat Sci. 2023 Jun; 70(2):145\u2013153. https://doi.org/10.1002/jmrs.673CPD Questions J Med Radiat Sci. 2023 Sep; 70(3): 229\u2013238. https://doi.org/10.1002/jmrs.676Edwards C, Perry R, Chester D, Childs J. Entrustable professional activities of graduate accredited General Medical Sonographers in Australia \u2013 industry perceptions. https://doi.org/10.1002/jmrs.696CPD Questions J Med Radiat Sci. 2023 Sep; 70(3): 292\u2013300. https://doi.org/10.1002/jmrs.699Fong SC, Pandey R, Rajaretnam M, Delaibatiki M, Peel DNY. Routine prophylactic percutaneous endoscopic gastrostomy in head and neck cancers with bilateral neck irradiation: a regional cancer experience in New Zealand. https://doi.org/10.1002/jmrs.708CPD Questions"} +{"text": "In the below-listed manuscripts, a change has been made to the author list.https://doi.org/10.1093/ofid/ofz061https://doi.org/10.1093/ofid/ofy182https://doi.org/10.1093/ofid/ofy289"} +{"text": "ACR Open Rheumatol. 2023 Mar;5(3):114. doi: 10.1002/acr2.11528Yokochi R, Hagino N. Clinical Images: Massive soft\u2010tissue calcification in primary Sjogren syndrome. Dr. Yokochi's first name was misspelled as Rtisuko in the published version. The correct spelling is Ritsuko.We apologize for this error."} +{"text": "This article has been retracted: Oncotarget is retracting this article at the request of the Scientific Integrity office. Similarities were found between images in this article and in other articles:https://doi.org/10.1186/s12885-017-3132-91. BMC Cancer (2017), https://doi.org/10.1155/2017/61364012. Disease Markers (2017), https://pubpeer.com/publications/1D3EFEEE1E55BFC3B47D193AEFECE6).Reader concerns were initially reported in a Pubpeer thread (Image analysis performed by the Oncotarget Scientific Integrity Office confirmed the findings of fraudulent image duplication. All authors have agreed to this retraction.113977-113986. https://doi.org/10.18632/oncotarget.23048Original article: Oncotarget. 2017; 8:113977\u2013113986."} +{"text": "Correction: BMC Endocr Disord 23, 81 (2023).10.1186/s12902-023-01339-wFollowing publication of the original article , the autThe correct funding statement should be:The study was supported by the European Regional Development Fund - Project ENOCH (No. CZ.02.1.01/0.0/0.0/16_019/0000868).The original article has been"} +{"text": "BMC Medical Informatics and Decision Making (2023) 23:8410.1186/s12911-023-02177-5The original publication of this article contained an incorrect ethics code. The incorrect and correct information is listed in this correction article. The original article has been updated.Incorrect: IR.ABADANUMS. REC.1401.048Correct: IR.ABADANUMS. REC.1401.101"} +{"text": "This article has an addendum: Because this was an autopsy-based analysis, autopsy/donation consent was obtained.42837-42842. https://doi.org/10.18632/oncotarget.10034Original article: Oncotarget. 2016; 7:42837\u201342842."} +{"text": "This article has been corrected: In 86592-86603. https://doi.org/10.18632/oncotarget.21246Original article: Oncotarget. 2017; 8:86592\u201386603."} +{"text": "Correction: BioData Mining 14, 29 (2021)https://doi.org/10.1186/s13040-021-00261-yhttps://www.proteinatlas.org/). Therefore, the results in this article are reliable. To make this research repeatable, the following has provided a download link for pathological images.Following publication of the original article , the autThe patient ID of normal liver tissue is 3402. link:https://www.proteinatlas.org/ENSG00000170231-FABP6/tissue/liver#img;FABP6 normalThe liver cancer patient ID is 3334, link:https://www.proteinatlas.org/ENSG00000170231-FABP6/pathology/liver+cancer#img.FABP6 liver cancerTherefore, the correct Fig."} +{"text": "J Clin Invest. 2022;132(22):e160101. https://doi.org/10.1172/JCI160101Original citation: J Clin Invest. 2023;133(13):e173145. https://doi.org/10.1172/JCI173145Citation for this corrigendum: neo2/neo2 and Foxn11089/1089 hypoplastic lobes were duplicates. The correct figure part is below. The HTML and PDF files have been updated online.In the original version of The authors regret the error."} +{"text": "The second author\u2019s name is spelled incorrectly. The correct name is: Rossana Pulcinelli V. Franciscohttps://doi.org/10.1371/journal.pone.0261492The correct citation is: Gon\u00e7alves BMM, Francisco RPV, Rodrigues AS (2021) Maternal mortality associated with COVID-19 in Brazil in 2020 and 2021: Comparison with non-pregnant women and men. PloS ONE 16(12): e0261492."} +{"text": "The third author\u2019s name is spelled incorrectly. The correct name is: Pratap Vydyam.https://doi.org/10.1371/journal.pone.0125358The correct citation is: Badugu SB, Nabi SA, Vydyam P, Laskar S, Bhattacharyya S, Bhattacharyya MK (2015) Identification of Plasmodium falciparum DNA Repair Protein Mre11 with an Evolutionarily Conserved Nuclease Function. PloS ONE 10(5): e0125358."} +{"text": "This article has an addendum: Informed patient consent was obtained.3800-3804. https://doi.org/10.18632/oncotarget.27771Original article: Oncotarget. 2020; 11:3800\u20133804."} +{"text": "This article has been corrected: In n = 2 but only one is reported here. The corrected 31134-31150. https://doi.org/10.18632/oncotarget.5290Original article: Oncotarget. 2015; 6:31134\u201331150."} +{"text": "Correction to: BMC Surgery 10.1186/s12893-023-02070-y.Following publication of the original article , in thiswww.editage.jp) for English language editing. The authors are grateful to JPSKAKENHI #22K16548.The authors thank Editage ("} +{"text": "Phil. Trans. R. Soc. B377, 20200498 (Published online 7 March 2022). (https://doi.org/10.1098/rstb.2020.0498)An incorrect version of The corrected figure is shown below and has also been corrected on the publiser's website."} +{"text": "This article has been corrected: In 44326-44334. https://doi.org/10.18632/oncotarget.17872Original article: Oncotarget. 2017; 8:44326\u201344334."} +{"text": "This article has been corrected: Due to errors during figure assembly, the control images used in the bottom row of 11640-11651. https://doi.org/10.18632/oncotarget.3449Original article: Oncotarget. 2015; 6:11640\u201311651."} +{"text": "Correction: BMC Neurol 23, 216 (2023)https://doi.org/10.1186/s12883-023-03261-zhttps://doi.org/10.3389/fneur.2019.01098) was incomplete and lacked DOI.Following publication of the original article , the autThe correct figures are presented below.The original article has been"} +{"text": "This article has been corrected: In 68365-68380. https://doi.org/10.18632/oncotarget.20190Original article: Oncotarget. 2017; 8:68365\u201368380."} +{"text": "This article has an addendum: Informed patient consent was obtained for publication of this case report.9613-9617. https://doi.org/10.18632/oncotarget.7386Original article: Oncotarget. 2016; 7:9613\u20139617."} +{"text": "This article has an addendum: Research funding was obtained from the Cleveland Clinic Florida, Maroone Cancer Center, Malignant Hematology Research Fund.384-394. https://doi.org/10.18632/oncotarget.28415Original article: Oncotarget. 2023; 14:384\u2013394."} +{"text": "This article has an addendum: Informed patient consent was obtained from all patients.45005-45009. https://doi.org/10.18632/oncotarget.6528Original article: Oncotarget. 2015; 6:45005\u201345009."} +{"text": "Reference 17 is not correctand should be as follows:BlanazsA.; RyanA.J.; ArmesS.P.Predictive phasediagrams for RAFT aqueous dispersionpolymerization: Effect of block copolymer composition, molecular weight,and copolymer concentration.Macromolecules2012, 45, 5099\u22125107.10.1021/ma301059r(17)"} +{"text": "Adv. Sci. 2023, 10, 2205862https://doi.org/10.1002/advs.202205862In the originally published article there are errors in Figure"} +{"text": "This article has been corrected: In 32639-32654. https://doi.org/10.18632/oncotarget.15947Original article: Oncotarget. 2017; 8:32639\u201332654."} +{"text": "Interdisciplinary CardioVascular and Thoracic Surgery.Upon original publication, the below-listed manuscripts were included into Volume 36, Issue 7, instead of Volume 37, Issue 1 of The Publisher apologizes for this error, which has now been corrected.https://doi.org/10.1093/icvts/ivad070https://doi.org/10.1093/icvts/ivad112https://doi.org/10.1093/icvts/ivad113"} +{"text": "This article has been corrected: In 76920-76933. https://doi.org/10.18632/oncotarget.12729Original article: Oncotarget. 2016; 7:76920\u201376933."} +{"text": "Details of correction: update email address for corresponding author.Existing text:sierksma@kennisinstituutbier.nlCorrected text should read:info@beerandhealth.eu"} +{"text": "This article has an addendum: No external funding was obtained.464-475. https://doi.org/10.18632/oncotarget.28213Original article: Oncotarget. 2022; 13:464\u2013475."} +{"text": "J Clin Invest. 2023;133(11):e168121. https://doi.org/10.1172/JCI168121Original citation: J Clin Invest. 2023;133(13):e172916. https://doi.org/10.1172/JCI172916Citation for this corrigendum: After the publication of this Review, the authors became aware of errors in"} +{"text": "J Clin Invest. 2021;131(1):e136779. https://doi.org/10.1172/JCI136779Original citation: J Clin Invest. 2023;133(10):e171901. https://doi.org/10.1172/JCI171901Citation for this corrigendum: The authors recently became aware of an inadvertent error in The authors regret the error."} +{"text": "This article has an addendum: Patient consent was required and obtained.33043-33049. https://doi.org/10.18632/oncotarget.25984Original article: Oncotarget. 2018; 9:33043\u201333049."} +{"text": "This article has been corrected: In 48110-48125. https://doi.org/10.18632/oncotarget.18262Original article: Oncotarget. 2017; 8:48110\u201348125."} +{"text": "Scientific Reports 10.1038/s41598-022-09445-w, published online 31 March 2022Correction to: The Data Availability section in the original version of this Article was omitted.It now appears as below:https://www.ncbi.nlm.nih.gov/nuccore/ON637243 and https://www.ncbi.nlm.nih.gov/nuccore/ON637244.\u201d\u201cThe Mus musculus transgenic K18-hACE2_PrlmnJ sequences are available here The original Article has been corrected."} +{"text": "This article has been corrected: In 944-959. https://doi.org/10.18632/oncotarget.28261Original article: Oncotarget. 2022; 13:944\u2013959."} +{"text": "This article has been corrected: In 2919-2929. https://doi.org/10.18632/oncotarget.27679Original article: Oncotarget. 2020; 11:2919\u20132929."} +{"text": "This article has an addendum: No external funding was obtained for this article.153-172. https://doi.org/10.18632/oncotarget.28367Original article: Oncotarget. 2023; 14:153\u2013172."} +{"text": "Proc. R. Soc. B290, 20222571. (Published online 1 March 2023). (https://doi.org/10.1098/rspb.2022.2571)n = 14 and n = 7; see This file contains a corrected version of This has been corrected on the publisher's website."} +{"text": "This article has an addendum: No funding was used for this publication.351-357. https://doi.org/10.18632/oncotarget.28406Original article: Oncotarget. 2023; 14:351\u2013357."} +{"text": "This article has an addendum: No external funding was provided for this research.178-187. https://doi.org/10.18632/oncotarget.28376Original article: Oncotarget. 2023; 14:178\u2013187."} +{"text": "Adv. Sci. 2023, 10, 230028810.1002/advs.202300288DOI: In the originally published article there are errors in Figure"} +{"text": "There was an error in Reference 117. The correct version of this Reference is available below.Attal Y, Maess B, Friederici A, David O (2012) Head models and dynamic causal modeling of subcortical activity using magnetoencephalographic/electroencephalo\u200bgraphicdata. Rev Neurosci 23: 85\u201395. doi: 10.1515/rns.2011.056."} +{"text": "C. muridarum Replication In Vitro, but Not In Vivo. The word \"Controlling\" in the title was misspelled. The correct title is: Perforin Is Detrimental to Controlling C. muridarum Replication In Vitro, but Not In Vivo. PLoS ONE 8(5): e63340. doi:10.1371/journal.pone.0063340. The correct citation is: Johnson RM, Kerr MS, Slaven JE (2013) Perforin Is Detrimental to Controlling"} +{"text": "There were errors in the Author Contributions. The correct contributions are: Conceived and designed the experiments: RWC. Analyzed the data: IMG SH. Contributed reagents/materials/analysis tools: SH. Wrote the paper: IMG RWC SH. Performed Numerical Simulations: IMG. Developed theory: IMG RWC SH."} +{"text": "There was information omitted from the Author Contributions. The correct version of this section is available below.Conceived and designed the experiments: DL CCV. Performed the experiments: DL EG JC. Analyzed the data: DL. Contributed reagents/materials/analysis tools: DL JKEM JC EG CB CCV. Wrote the paper: DL JKEM CCV. Logistics: DL JKEM EG CB. Permits: DL. Data collection: DL EG JC."} +{"text": "Biological data set, two equations are missing. The corrected text can be viewed here: http://plosone.org/corrections/pone.0046237.e001.cn.tifIn the Materials and Methods section, under the heading"} +{"text": "In the E-mail list, guoyahong7225@163.com (YG) should be in front of maozuguo@hit.edu.cn (MG). The E-mail list should read:\"*E-mail: guoyahong7225@163.com (YG); maozuguo@hit.edu.cn (MG); yufei.huang@utsa.edu (YH)\"Posted by the authors of this article."} +{"text": "The enhanced recovery after surgery (ERAS) programme is a multimodal evidence-based approach to surgical care which begins in the preoperative setting and extends through to patient discharge in the postoperative period. The primary components of ERAS include the introduction of preoperative patient education; reduction in perioperative use of nasogastric tubes and drains; the use of multimodal analgesia; goal-directed fluid management; early removal of Foley catheter; early mobilization, and early oral nutrition. The ERAS approach has gradually evolved to become the standard of care in colorectal surgery and is presently being used in other specialty areas such as vascular surgery. Currently there is little evidence available for the implementation of ERAS in this field. We plan to conduct a systematic review of this literature with a view to incorporating ERAS principles into the management of major elective vascular surgery procedures.Register of Controlled Trials . Searches will be performed with no year or language restrictions. For inclusion, studies must look at adult patients over 18 years. Major elective vascular surgery includes carotid, bypass, aneurysm and amputation procedures. Studies must have evaluated usual care against an ERAS intervention in the preoperative, perioperative or postoperative period of care. Primary outcome measures are length of stay, decreased complication rate, and patient satisfaction or expectations. Only randomized controlled trials will be included.We will search EMBASE , Medline , and Cochrane Central Most ERAS approaches have been considered in the context of colorectal surgery. Given the increasing use of multiple yet different aspects of this pathway in vascular surgery, it is timely to systematically review the evidence for their independent or combined outcomes, with a view to implementing them in this clinical setting. Results from this review will have important implications for vascular surgeons, anaesthetists, nurses, and other health care professionals when making evidenced-based decisions about the use of ERAS in daily practice. The enhanced recovery after surgery (ERAS) programme is a multimodal evidence-based approach to surgical care which begins in the preoperative setting and extends through to patient discharge in the postoperative period. This interprofessional, goal-directed pathway, involves surgeons, nurses, anaesthetists, and physiotherapists. It aims to accelerate the recovery of surgical patients while decreasing complications and reducing hospital length of stay. Since the early 1990s, the ERAS approach has gradually evolved to become the standard of care in colorectal surgery-15. EvidThe use of ERAS protocols is reported in other surgical specialty areas, such as vascular,20, cardWhile there are a handful of general reviews on the topic of ERAS in vascular surgery,31,32 thEligibility: We will include studies of adult patients over the age of 18 who have undergone major elective vascular surgery and received an ERAS intervention. For this review major elective vascular surgery is defined as a carotid, aneurysm, bypass, or amputation procedure (see Appendix I). Studies must have evaluated the use of one or more ERAS intervention against usual care for patients , Medline , and Cochrane Central Register of Controlled Trials . The database search will be supplemented by searching for grey literature . Specifically, we will search online , trial registers , and conference abstracts. Searches will be performed with no year or language restrictions. Relevant non-English language articles will be translated. Literature search strategies will be developed by combining medical subject headings (MeSH terms) and appropriate wildcards. If necessary we will contact study authors to identify additional studies. The literature search will be conducted by an information specialist (LP) and peer-reviewed by another information specialist using Peer Review of Electronic Search Strategy (PRESS). In addiA pilot test of 50 randomly selected citations will be conducted by all authors to verify the inclusion/exclusion criteria. Subsequently, all studies (citations and full text) will be reviewed by two reviewers independently. Conflicts will be resolved by team discussion.Data from included studies will be abstracted in duplicate by two reviewers using a data collection form. The data collection form will include:a. patient characteristics b. study characteristics c. details of the intervention d. details of the comparator e. primary outcome results .The data abstraction form will be pilot tested on a random sample of studies to ensure high inter-rater agreement between reviewers.Two reviewers will independently assess each selected study for the strengths of the research methods and results using the Cochrane Risk of Bias Tool. This to2 statistic2. Aortic Aneurysm, Abdominal/su3. Aortic Aneurysm, Thoracic/su4. Endarterectomy, Carotid/5. Endarterectomy/6. (aneurysm adj repair$).tw.7. (aneurysm adj surg$).tw.8. (aort$ adj aneurysm$).mp.9. (aort$ adj3 repair$).tw.10. (aort$ adj3 bypass$).tw.11. (aort$ adj3 by-pass$).tw.12. (aort$ adj reconstruction$).tw.13. (aort$ adj surg$).tw.14. (arter$ adj aneurysm).tw.15. (arter$ adj bypass$).tw.16. (arter$ adj by-pass$).tw.17. (arter$ adj repair$).tw.18. (arter$ adj surg$).tw.19. (axillo$ adj2 bypass$).tw.20. (axillo$ adj2 by-pass$).tw.21. ABF.tw.22. AUI.tw.23. AKA.tw.24. (AAA adj repair$).tw.25. (AAA adj surg$).tw.26. (bypass$ adj surg$).tw.27. (by-pass$ adj surg$).tw.28. BKA.tw.29. (carotid adj surg$).tw.30. CEA.tw.31. endarterectom$.tw.32. (endovascular adj repair$).tw.33. (endovascular adj surg$).tw.34. (femor$ adj aneurysm).tw.35. (femor$ adj repair$).tw.36. (femor$ adj2 crossover$).tw.37. (femor$ adj2 cross-over$).tw.38. (femor$ adj3 bypass$).tw.39. (femor$ adj3 by-pass$).tw.40. (foot adj amputat$).tw.41. \"ilio femor$ bypass$\".tw.42. \"ilio femor$ by-pass$\".tw.43. .tw.44. .tw.45. .tw.46. .tw.47. .tw.48. .tw.49. .tw.50. (knee adj1 amputat$).tw.51. (leg adj revascular$).tw.52. \"popliteal pedal bypass$\".tw.53. \"popliteal pedal by-pass$\".tw.54. profundoplast$.tw.55. .tw.56. .tw.57. .tw.58. .tw.59. (symes adj amputat$).tw.60. (tibia$ adj bypass$).tw.61. (tibia$ adj by-pass$).tw.62. .tw.63. (TAA adj repair$).tw.64. (TAA adj surg$).tw.65. TEVAR.tw.66. (vascular adj bypass).tw.67. (vascular adj by-pass).tw.68. (vascular adj surg$).tw.69. (vascular adj repair$).tw.70. (vascular adj reconstruction$).tw.71. or/1-7072. Chewing Gum/[Enhanced Recovery]73. Early Ambulation/74. Exercise Therapy/75. Heating/76. Intraoperative Care/mt77. Preoperative Care/mt78. Perioperative Care/mt79. Postoperative Care/mt80. Patient Education as Topic/81. Surgical Procedures, Minimally Invasive/82. exp Anesthesia/83. an?esthesia.tw.84. an?esthetic?.tw.85. (accelerat$ adj2 mobil$).tw.86. (accelerat$ adj2 ambulat$).tw.87. .tw.88. (accelerat$ adj2 feed$).tw.89. (accelerat$ adj2 nutrition$).tw.90. (accelerat$ adj2 eat$).tw.91. (accelerat$ adj2 rehab$).tw.92. (chew$ adj1 gum?).tw.93. (client$ adj educat$).tw.94. (client$ adj teach$).tw.95. (client$ adj counsel$).tw.96. (client$ adj expectation$).tw.97. .tw.98. .tw.99. (earl$ adj2 mobil$).tw.100. (earl$ adj2 ambulat$).tw.101. .tw.102. (earl$ adj2 feed$).tw.103. (earl$ adj2 nutrition$).tw.104. (earl$ adj2 eat$).tw.105. (earl$ adj2 rehab$).tw.106. (enhanced adj recover$).tw.107. ERAS.tw.108. (fast adj tract$).tw. (12)109. (fast adj track$).tw. (1787)110. (heat$ adj2 patient$).tw. (440)111. intraoperative.mp. and (intravenous adj fluid?).tw. (207)112. intraoperative.mp. and (IV adj fluid?).tw. (75)113. intraoperative.mp. and Infusions, Intravenous/(856)114. intraoperative.mp. and Intubation, Gastrointestinal/(123)115. intraoperative.mp. and (NG adj tube?).tw. (5)116. intraoperative.mp. and (nasogastric adj tube?).tw. (108)117. intraoperative.mp. and (fluid? adj1 restrict$).tw. (59)118. intraoperative.mp. and .tw. (35)119. intraoperative.mp. and Anti-Inflammatory Agents, Non-Steroidal/241)120. intraoperative.mp. and NSAID?.tw.121. .mp.122. .mp.123. (intraoperative and narcotic?).mp.124. (intraoperative and (fluid adj therap$)).mp.125. intraoperative.mp. and (fluid adj manag$).tw.126. intraoperative.mp. and (electrolyte$ adj manag$).tw.127. (intraoperative and (pain adj manage$)).mp.128. (intraoperative and (vein adj thrombos?s)).mp.129. ((intraoperative adj care) and enhanced).tw.130. ((intraoperative adj care) and accelerat$).tw.131. ((intraoperative adj care) and early).tw.132. intra-operative.mp. and (intravenous adj fluid?).tw.133. intra-operative.mp. and (IV adj fluid?).tw.134. intra-operative.mp. and Infusions, Intravenous/135. intra-operative.mp. and Intubation, Gastrointestinal/136. intra-operative.mp. and (NG adj tube?).tw.137. intra-operative.mp. and (nasogastric adj tube?).tw.138. intra-operative.mp. and (fluid? adj1 restrict$).tw.139. intra-operative.mp. and .tw.140. intra-operative.mp. and Anti-Inflammatory Agents, Non-Steroidal/141. intra-operative.mp. and NSAID?.tw.142. .mp.143. .mp.144. (intra-operative and narcotic?).mp.145. (intra-operative and (fluid adj therap$)).mp.146. intra-operative.mp. and (fluid adj manag$).tw.147. intra-operative.mp. and (electrolyte$ adj manag$).tw.148. (intra-operative and (pain adj manage$)).mp.149. (intra-operative and (vein adj thrombos?s)).mp.150. ((intra-operative adj care) and enhanced).tw.151. ((intra-operative adj care) and accelerat$).tw.152. ((intra-operative adj care) and early).tw.153. .tw.154. .tw.155. (patient$ adj educat$).tw.156. (patient$ adj teach$).tw.157. (patient$ adj counsel$).tw.158. (patient$ adj expectation$).tw.159. ((perioperative adj care) and enhanced).tw.160. ((perioperative adj care) and accelerat$).tw.161. ((perioperative adj care) and early).tw.162. ((peri-operative adj care) and enhanced).tw.163. ((peri-operative adj care) and accelerat$).tw.164. ((peri-operative adj care) and early).tw.165. postoperative.mp. and (regular adj diet?).tw.166. postoperative.mp. and .tw.167. postoperative.mp. and Enteral Nutrition/168. (postoperative and catheter?).mp.169. (postoperative and (fluid adj therap$)).mp.170. postoperative.mp. and (fluid adj manage$).tw.171. postoperative.mp. and (electrolyte$ adj manag$).tw.172. .mp.173. (postoperative and opioid?).mp.174. (postoperative and opiat$).mp.175. .mp.176. postoperative.mp. and Anti-Inflammatory Agents, Non-Steroidal/177. postoperative.mp. and NSAID?.tw.178. (postoperative and (pain adj manage$)).mp.179. postoperative.mp. and (care adj map?).tw.180. postoperative.mp. and (care adj plan$).tw.181. postoperative.mp. and (treatment adj plan$).tw.182. ).mp.183. (postoperative and (care adj path$)).tw.184. ).mp.185. (postoperative and (case adj management)).mp.186. (postoperative and (patient adj discharge)).mp.187. (postoperative and (discharge adj plan$)).mp.188. (postoperative and (vein adj thrombos?s)).mp.189. (postoperative and antiemetic?).mp.190. (postoperative and anti-emetic?).mp.191. (postoperative and ileus).mp.192. ((postoperative adj care) and enhanced).tw.193. ((postoperative adj care) and accelerat$).tw.194. ((postoperative adj care) and early).tw.195. post-operative.mp. and (regular adj diet?).tw.196. post-operative.mp. and .tw.197. post-operative.mp. and Enteral Nutrition/198. (post-operative and catheter?).mp.199. (post-operative and (fluid adj therap$)).mp.200. post-operative.mp. and (fluid adj manage$).tw.201. post-operative.mp. and (electrolyte$ adj manag$).tw.202. .mp.203. (post-operative and opioid?).mp.204. (post-operative and opiat$).mp.205. .mp.206. post-operative.mp. and Anti-Inflammatory Agents, Non-Steroidal/207. post-operative.mp. and NSAID?.tw.208. (post-operative and (pain adj manage$)).mp.209. post-operative.mp. and (care adj map?).tw.210. post-operative.mp. and (care adj plan$).tw.211. post-operative.mp. and (treatment adj plan$).tw.212. ).mp.213. (post-operative and (care adj path$)).tw.214. ).mp.215. (post-operative and (case adj management)).mp.216. (post-operative and (patient adj discharge)).mp.217. post-operative.mp. and (discharge adj plan$).tw.218. (post-operative and (vein adj thrombos?s)).mp.219. (post-operative and antiemetic?).mp.220. (post-operative and anti-emetic?).mp.221. (post-operative and ileus).mp.222. ((post-operative adj care) and enhanced).tw.223. ((post-operative adj care) and accelerat$).tw.224. ((post-operative adj care) and early).tw.225. (preoperative and fasting).mp.226. preoperative.mp. and Anti-Inflammatory Agents, Non-Steroidal/227. preoperative.mp. and NSAID?.tw.228. preoperative.mp. and carbohydrate$.tw.229. (preoperative and probiotic?).mp.230. (preoperative and pro-biotic?).mp.231. preoperative.mp. and hydrat$.tw.232. preoperative.mp. and dehydrat$.tw.233. preoperative.mp. and de-hydrat$.tw.234. preoperative.mp. and stress.tw.235. ((preoperative adj care) and enhanced).tw.236. ((preoperative adj care) and accelerat$).tw.237. ((preoperative adj care) and early).tw.238. (pre-operative and fasting).mp.239. pre-operative.mp. and Anti-Inflammatory Agents, Non-Steroidal/240. pre-operative.mp. and NSAID?.tw.241. pre-operative.mp. and carbohydrate$.tw.242. (pre-operative and probiotic?).mp.243. (pre-operative and pro-biotic?).mp.244. pre-operative.mp. and hydrat$.tw.245. pre-operative.mp. and dehydrat$.tw.246. pre-operative.mp. and de-hydrat$.tw.247. pre-operative.mp. and stress.tw.248. ((pre-operative adj care) and enhanced).tw.249. ((pre-operative adj care) and accelerat$).tw.250. ((pre-operative adj care) and early).tw.251. (rapid$ adj2 recover$).tw.252. (rapid adj2 mobil$).tw.253. (rapid adj2 ambulat$).tw.254. .tw.255. (rapid adj2 feed$).tw.256. (rapid adj2 nutrition$).tw.257. (rapid adj2 eat$).tw.258. (rapid adj2 rehab$).tw.259. (warm$ adj patient$).tw.260. or/72-259261. 71 and 260262. exp Animals/ not 263. 261 not 262 [ Removing Animal Studies ]264. randomized controlled trial.pt. [ RCT filter - validated - optimized ]265. randomized.mp.266. placebo.mp.267. or/264-266268. 263 and 267ERAS: Enhanced recovery after surgery; AAA: Abdominal aortic aneurysm; EVAR: Endovascular aneurysm repair; AUI: Aorto-uni-iliac; ABF: Aorto-Bifemoral bypass; NSAIDs: Non-Steroidal Anti-Inflammatory drugs; MeSH: Medical subject headings.There are no competing interests.LGC collaborated in the design of the study and drafted the protocol. OR conceived of the study, collaborated in the design of the study, and edited the protocol. EG collaborated in the design of the study and edited the protocol. ACT collaborated in the design of the study and edited the protocol. LP developed the search strategy and edited the protocol. CS collaborated in the design of the study and edited the protocol. TM collaborated in the design of the study and edited the protocol. All authors read and approved the final manuscript.This systematic review is funded by the Department of Surgery, St. Michael\u2019s Hospital. ACT is funded by a Canadian Institutes for Health Research/Drug Safety and Effectiveness Network New Investigator Award in Knowledge Synthesis."} +{"text": "It was published in 2012, so it contains recent information.http://rkt.chem.ox.ac.uk/projects/biosurfactants.htmlThis academic research page provides a good overview of biosurfactants.http://www.prweb.com/releases/2012/11/prweb10158903.htmWhile the full report is available only for a fee, this page hints at the importance of biosurfactants commercially.http://www.ideaconnection.com/crowdfunding/isolation-and-characterization-of-biosurfactant-produ-00041.htmlThis page on the IdeaConnection website proposes research on bacterial biosurfactants.http://envismadrasuniv.org/Bioremediation/pdf/Biosurfactants%20and%20oil%20bioremediation.pdfThis page links to a comprehensive review article on the role of biosurfactants in bioremediation.http://www.amb-express.com/content/1/1/9/abstractThis is a typical example in which an alkane-degrading pseudomonad makes a rhamnolipid surfactant that aids in solubilizing and degrading the alkanes.http://www.columbia.edu/cu/iucrc/research.shtmlThis list of research at a university centre describes numerous projects on surfactants, including biosurfactants.http://www.dfo-mpo.gc.ca/science/publications/microbes/index-eng.htmlThis page contains a reprinting of a document from the American Academy of Microbiology that answers frequently asked questions about oil spills, largely from the perspective of how microorgansisms are impacted and impact the spill.http://www.biosurf.ugent.be/Biosurfactantia.htmThis introduction to biosurfacants contains text and shows chemical structures of characterized biosurfactants.http://nopr.niscair.res.in/bitstream/123456789/4804/1/JSIR%2065(2)%2091-115.pdfThis review arrticle contains information on more than 250 patents relevant to biosurfactants.http://www.iisc.ernet.in/currsci/jul10/articles19.htmThis web review article contains detailed information and more than 160 references.http://gradworks.umi.com/35/50/3550595.htmlThis page links to a dissertation that describes the multiple environmental roles of microbial biosurfactants. Examples include their role in biodegradation and aid in swarming motility.http://www.igb.fraunhofer.de/en/competences/molecular-biotechnology/white-biotechnology/biosurf.htmlThis page describes an institutional programme to develop biosurfactants for industrial uses.http://www.tau.ac.il/lifesci/departments/biotech/members/rosenberg/rosenberg.htmlThis personal web page contains links to many papers and patents pertaining to biosurfactants."} +{"text": "The first author's name was incorrectly given in the byline and citation.The correct name is K. Anne-Isola NekarisNycticebus spp.) on Web 2.0 Sites. PLoS ONE 8(7): e69215. doi:10.1371/journal.pone.0069215. The correct citation is: Nekaris KA-I, Campbell N, Coggins TG, Rode EJ, Nijman V (2013) Tickled to Death: Analysing Public Perceptions of \u2018Cute\u2019 Videos of Threatened Species (Slow Lorises \u2013"} +{"text": "There was an error in the third author's name. Srivathsa S. Magadi is correct. The correct citation is: Kulkarni V, Khadilkar RJ, Magadi SS, Inamdar MS (2011) Asrij Maintains the Stem Cell Niche and Controls Differentiation during Drosophila Lymph Gland Hematopoiesis. PLoS ONE 6(11): e27667. doi:10.1371/journal.pone.0027667"} +{"text": "Please see the corrected Figure 1 here: http://plosone.org/corrections/pone.0071659.t001.cn.tifThe P-Values in Table 1 were erroneously altered. Please see the corrected Table 1 here:"} +{"text": "Mycobacterium tuberculosis Infection of Domesticated Asian Elephants, Thailand . The article has been corrected online (http://wwwnc.cdc.gov/eid/article/16/12/10-0862_article).An online Technical Appendix was omitted from the article"} +{"text": "The name of the first author is incorrect. The correct name is: Aditi J. Ravindranath.The correct Citation is: Ravindranath AJ, Cadigan KM (2014) Structure-Function Analysis of the C-clamp of TCF/Pangolin in Wnt/\u00df-catenin Signaling. PLoS ONE 9(1): e86180. doi:10.1371/journal.pone.0086180."} +{"text": "There was an error in the fourth author's name. The correct name is Gabri\u00eblle H.S. Buitendijk. The correct citation is van Beveren NJM, Krab LC, Swagemakers S, Buitendijk GHS, Boot E, et al. (2012) Functional Gene-Expression Analysis Shows Involvement of Schizophrenia-Relevant Pathways in Patients with 22q11 Deletion Syndrome. PLoS ONE 7(3): e33473. doi:10.1371/journal.pone.0033473"} +{"text": "The image for Figure 2 is incorrect. Please view the correct image here:http://www.plosntd.org/corrections/pntd.0001885.g002.cn.tif"} +{"text": "The name of author A.A.T. Simone Reinders contained an error in the citation. The correct citation should read: Reinders AATS, Willemsen ATM, Vos HPJ, den Boer JA, Nijenhuis ERS (2012) Fact or Factitious? A Psychobiological Study of Authentic and Simulated Dissociative Identity States. PLoS ONE 7(6): e39279. doi:10.1371/journal.pone.0039279"} +{"text": "AbstractSesioctonus (Braconidae: Agathidinae) are described and illustrated, i.e., Sesioctonus huggerti, Sesioctonus wayquecha, and Sesioctonus bina. Two new Peruvian species records for Sesioctonus are reported: Sesioctonus longinoi and Sesioctonus diazi. A revised key to all known species of Sesioctonus is presented.Three new species of Sesioctonus Viereck, 1912 is a Neotropical genus of Agathidinae which for which the biology is largely unknown, only Sesioctonus parathyridis is recorded as a larval parasitoid of Parathyris perspicilla Stall (Lepidoptera: Arctiidae) (Viereck 1912). Sesioctonus and included twenty six species and Morphological terminology follows that of Unless otherwise stated specimens are deposited in the Natural History Museum, University of San Marcos, Lima, Peru (MUSM), with duplicates deposited in the Hymenoptera Institute Collection at the University of Kentucky, USA (HIC).Sesioctonus are restricted to the Neotropical realm of the New World and may be distinguished from all other agathidine braconids with the following combination of characters: Mesoscutum smooth, lacking notauli; tarsal claws simple, lacking a basal claw; hind coxal cavities open, sharing a common opening with the metasomal foramen.Members of Sulca & Sharkeysp. n.urn:lsid:zoobank.org:act:A198E0BC-7DFE-42CB-B5AD-9DF7E63597E6http://species-id.net/wiki/Sesioctonus_huggertiSesioctonus by the following suite of characters: Interantennal space lacking longitudinal keel, epicnemial carinae straight medially.Distinguished from all other known species of Length. Length of body, excluding ovipositor, 5 mm.\u2640 Head. Flagellum with 30 flagellomeres. Interantennal space lacking longitudinal keel. Antennal sockets moderately excavated. Face with median longitudinal carina. Gena not expanded posteroventrally. Occipital tubercles present. Occiput not excavated. Mandible concave. Outer tooth of mandible not longer than inner tooth. Maxillary palpus with 4 palpomeres. Third and fourth labial palpomeres not fused. Mesosoma. Subpronope elongate-oval. Longitudinal carinae of scutellar depression absent. Scutellum convex. Median areola of metanotum smooth, with median longitudinal carina, and with lateral carinae present and not meeting posteriorly. Propodeum convex. Median longitudinal carina of propodeum absent. Epicnemial carina complete, sharp, straight medially (between fore coxae). Hind femur 6 times as long as wide. (RS+M)a vein of fore wing incomplete. 3RSa vein of fore wing absent. 2\u20131A vein of hind wing not tubular. Cub vein of hind wing not tubular. Metasoma. Median tergite of first metasomal segment without pair of lateral longitudinal carinae. Hind wing with 4 hamuli. First metasomal median tergite without depression posteriad spiracle. Length/width ratio of first metasomal median tergite 0.63. Ovipositor 4 mm.Color. Head melanic. Maxillary palpomeres melanic. Labial palpomeres melanic. Pronotum melanic. Mesoscutum yellowish orange. Scutellum yellowish orange. Metanotum yellowish orange. Propodeum melanic. Propleuron melanic. Mesopleuron yellowish orange. Metapleuron melanic. Fore coxa melanic. Fore trochanter melanic. Fore trochantellus melanic. Fore femur melanic. Fore tibia melanic. Fore tarsus melanic. Mid coxa melanic. Mid trochanter melanic. Mid trochantellus melanic. Midfemur melanic. Mid tibia melanic. Mid tarsus melanic. Hind coxa melanic. Hind trochanter melanic. Hind trochantellus melanic. Hind femur melanic. Hind tibia melanic. Hind tarsus melanic. Fore wing entirely infuscate. Stigma melanic. Hind wing entirely infuscate. First metasomal tergum melanic. Second metasomal tergum mePageBreaklanic. Third metasomal tergum melanic. Fourth metasomal tergum melanic. Fifth to eighth metasomal terga melanic. Ovipositor yellowish orange.\u2642 Unknown.Etymology. Named in honor of the late Lars Huggert who collected the type specimen.Holotype. PERU, Madre de Dios, Puerto Maldonado, 6\u201311.i.1984, L. Huggert Leg. .Known only from the type locality in Peru.Sulca & Sharkeysp. n.urn:lsid:zoobank.org:act:19BD24A0-162D-405A-8BF0-5CDA62C5FE86http://species-id.net/wiki/Sesioctonus_wayquechaSesioctonus by the following suite of characters: occipital tubercles absent, epicnemial carina completely absent, antennal socket not excavated, gena moderately expanded posteroventrally .Distinguished from all other known species of Length. Length of body, excluding ovipositor, 4.3\u20135.5 mm.\u2640 Head. Flagellum with 31 flagellomeres. Interantennal space lacking longitudinal keel. Antennal sockets not excavated. Face without median longitudinal carina. Gena moderately expanded posteroventrally. Occipital tubercles absent. Occiput not excavated. Mandible concave. Outer tooth of mandible not longer than inner tooth. Maxillary palpus with 5 palpomeres. Third and fourth labial palpomeres not fused. Mesosoma. Subpronope elongate-oval. Longitudinal carinae of scutellar depression absent. Scutellum convex. Median areola of metanotum smooth, without median longitudinal carina, and with lateral carinae present and meeting posteriorly. Propodeum convex. Median longitudinal carina of propodeum absent. Epicnemial carina completely absent. Fore tibial spines present. Mid tibia with 7 spines. Hind tibia with 15 spines. Hind femur 3.3\u20134 times as long as wide. (RS+M)a vein of fore wing complete. 3RSaPageBreak vein of fore wing absent. 2\u20131A vein of hind wing not tubular. Cub vein of hind wing not tubular. Metasoma. Median tergite of first metasomal segment without pair of lateral longitudinal carinae. Hind wing with 4 hamuli. First metasomal median tergite without depression posteriad spiracle. Length/width ratio of first metasomal median tergite 0.63. Ovipositor 0.5\u20130.6 mm.Color. Head melanic. Antenna melanic. Maxillary palpomeres yellowish orange. Labial palpomeres yellowish orange. Pronotum melanic. Mesoscutum melanic. Scutellum melanic. Metanotum melanic. Propodeum mostly yellowish orange with melanic spots. Propleuron mostly melanic with yellowish orange areas. Mesopleuron melanic. Metapleuron yellowish orange. Fore coxa yellowish orange. Fore trochanter yellowish orange. Fore trochantellus yellowish orange. Fore femur yellowish orange. Fore tibia melanic with yellowish orange ends. Fore tarsus melanic. Mid coxa yellowish orange. Mid trochanter yellowish orange. Mid trochantellus yellowish orange. Mid femur yellowish orange. Mid tibia melanic. Mid tarsus melanic. Hind coxa yellowish orange. Hind trochanter yellowish orange. Hind trochantellus yellowish orange. Hind femur yellowish orange. Hind tibia melanic with a yellow orange apical spot. Hind tarsus melanic. Fore wing entirely infuscate. Stigma melanic. Hind wing entirely infuscate. First metasomal tergum yellowish orange. Second metasomal tergum yellowish orange. Third metasomal tergum yellowish orange. Fourth metasomal tergum yellowish orange but median tergum melanic. Fifth to eighth metasomal terga melanic. Ovipositor yellowish orange.\u2642. As in the female (above).PageBreakEtymology. Named after the type locality, Wayquecha which means \u2018brother\u2019 in Quechua.Holotype. PERU. \u2640,Cusco, Wayquecha, 13\u00b011'21\"S, 71\u00b035'04\"W 2837m ,6\u2013 20.x.2007, C. Castillo. Leg.Paratypes: PERU: Cusco: 2\u2640\u2640,Wayquecha, 13\u00b011'21\"S, 71\u00b035'4\"W, 2837m, Malaise, 20.x.2007, C. Castillo Leg.; 3\u2640\u2640, 1\u2642,Wayquecha, 13\u00b010'31\"S, 71\u00b034'53\"W, 2692m, Malaise, 10.ix.2007, C. Castillo Leg.; \u2640, Wayquecha 13\u00b011'S, 71\u00b035'W, 2800m, sweep, 12.ix.2007, C. Castillo Leg; \u2642 Wayquecha, 13\u00b010'31\"S, 71\u00b034'53\"W, 2692m, Malaise, 22.x.2007, C. Castillo Leg.Known only from one locality in Peru.Sulca & Sharkeysp. n.urn:lsid:zoobank.org:act:5AE5EEED-ACF8-47D4-8189-531FFDBB1209http://species-id.net/wiki/Sesioctonus_binaSesioctonus by the following suite of characters: occiput not excavated, subpronope oval, median tergite of first metasomal segment with pair of lateral longitudinal carinae.Distinguished from all other known species of Length. Length of body, excluding ovipositor, 3.35 mm. Flagellum broken after flagellomere 28. Interantennal space with longitudinal rounded keel. Antennal sockets moderately excavated. Face without median longitudinal carina. Gena not expanded posteroventrally. Occipital tubercles present. Occiput not excavated. Mandible concave. Outer tooth of mandible not longer than inner tooth. Maxillary palpus with 4 palpomeres. Mesosoma. Subpronope oval. Longitudinal carinae of scutellar depression absent. Scutellum convex. Median areola of metanotum smooth, without median longitudinal carina, and with lateral carinae present and meeting posteriorly. Propodeum convex. Median longitudinal carina of propodeum present. Epicnemial carina complete, blunt, bilobed medially (between fore coxae). Fore tibial spines present. Mid tibia with 3 spines. Hind tibia with 12 spines. Hind femur 4 times as long as wide. (RS+M)a vein of fore wing incomplete. 3RSa vein of fore wing present. 2\u20131A vein of hind wing tubular. Cub vein of hind wing not tubular. Metasoma. Median tergite of first metasomal segment with pair of lateral longitudinal carinae. Hind wing with 3 hamuli. First metasomal median tergite without depression posteriad spiracle. Length width ratio of first metasomal median tergite 0.5. Ovipositor 1.68 mm.\u2640 Color. Head black and yellowish orange. Antenna melanic. Maxillary palpomeres yellowish orange. Labial palpomeres yellowish orange. Pronotum melanic. Mesoscutum melanic. Scutellum melanic. Metanotum melanic. Propodeum melanic. Propleuron melanic. Mesopleuron melanic. Metapleuron melanic. Fore coxa yellowish orange. Fore trochanter yellowish orange. Fore trochantellus yellowish orange. Fore femur yellowish orange. Fore tibia yellowish orange. Fore tarsus mostly yellowish orange, but apical tarsomere melanic. Mid coxa yellowish orange. Mid trochanter yellowish orange. MidPageBreak trochantellus yellowish orange. Mid femur yellowish orange. Mid tibia yellowish orange in basal half, melanic apically, or yellowish orange basally, otherwise melanic. Mid tarsus melanic. Hind coxa melanic. Hind trochanter melanic. Hind trochantellus melanic. Hind femur melanic. Hind tibia melanic in basal and apical third, yellowish orange medially. Hind tarsus melanic. Fore wing entirely infuscate. Stigma melanic. Hind wing entirely infuscate. First metasomal tergum melanic. Second metasomal tergum yellowish orange but median tergite melanic. Third metasomal tergum melanic. Fourth metasomal tergum melanic. Fifth to eighth metasomal terga melanic. Ovipositor yellowish orange.\u2642 unknownEtymology. Bina means \u2018wasp\u2019 in Shipibo, an indigenous language of the Peruvian Amazon.Holotype. \u2640, PERU, Cusco, Rocotal, 16.ix.2007, Sweep, C. Castillo Leg.Sesioctonus longinoi13\u00b003'23\"S, 71\u00b032'55\"W, 1520m, Malaise, 7.i.2009,C.Castillo. leg. \u2640,Cusco, San Pedro, 13\u00b003'23\"S, 71\u00b032'55\"W, 1520m, Malaise, C. Castillo. leg.\u2640, Cusco, Cos\u00f1ipata valley, San Pedro, Sesioctonus diazi12\u00b049,8'24\"S, 71\u00b005'55\"W, 507m, 11.x.2010. C.Castillo. leg. \u2640, Loreto, Alto Nanay, Albarenga north, 18M 0533605 9645694, 142m, C. Castillo leg.\u2640, Cusco, Reserva Comunal Amarakaeri, Rio Azul,"} +{"text": "There were errors in the Author Contributions. The correct contributions are: Conceived and designed the experiments: RE MSR RC HJB ITdA. Performed the experiments: RE MSR MB CF RMK. Analyzed the data: RE MSR TT YMS IR PL PK RC HJB ITdA. Contributed reagents/materials/analysis tools: TT PK RC HJB ITdA. Wrote the paper: RE MSR TT YMS IR PL PK RC HJB ITdA."} +{"text": "There were errors in the Author Contributions section. A correct version of the section is available below.Conceived and designed the experiments: FZ M.Lazzerini. Performed the experiments: FZ JAOG AC M.Lonardi MS PP DG CG. Analyzed the data: M.Lazzerini FZ. Contributed reagents/materials/analysis tools: FZ M.Lazzerini TN. Wrote the paper: M.Lazzerini, FZ."} +{"text": "Please view Dataset S1 here: http://www.plosbiology.org/corrections/pbio.1001105.s008.tif .and Dataset S2 here:"} +{"text": "Geranium sylvaticum. The correct citation is: Varga S, Nuortila C, Kyt\u00f6viita M-M (2013) Nectar Sugar Production across Floral Phases in the Gynodioecious Protandrous Plant Geranium sylvaticum. PLoS ONE 8(4): e62575. doi:10.1371/journal.pone.0062575. The words \"Gynodioecious\" and \"Protandrous\" in the article title were incorrectly italicized. The correct title is: Nectar Sugar Production across Floral Phases in the Gynodioecious Protandrous Plant"} +{"text": "T is erroneously cited as LMG 2670T.In the abstract of the above paper (doi:10.1099/ijs.0.040162-0), the type strain LMG 26707doi:10.1099/ijs.0.055368-0"} +{"text": "The second author's name should be: April W. Armstrong. The corrected citation is: Rehal B, Armstrong AW (2011) Health Outcome Measures in Atopic Dermatitis: A Systematic Review of Trends in Disease Severity and Quality-of-Life Instruments 1985 - 2010. PLoS ONE 6(4): e17520. doi:10.1371/journal.pone.0017520. The corrected Author Contributions are: Conceived and designed the experiments: AWA BR. Analyzed the data: AWA BR. Contributed reagents/materials/analysis tools: AWA BR. Wrote the paper: AWA BR."} +{"text": "There was an error in the third author's name.The correct name is: Tirtha K. Das.The correct citation is: Fu S, Yang Y, Das TK, Yen Y, Zhou B-s, et al. (2012) \u03b3-H2AX Kinetics as a Novel Approach to High Content Screening for Small Molecule Radiosensitizers. PLoS ONE 7(6): e38465. doi:10.1371/journal.pone.0038465 The correct Author Contributions are: Conceived and designed the experiments: SF TKD Y. Yen RC JK. Performed the experiments: SF Y. Yang TKD BZ JK. Analyzed the data: SF TKD MO MMZ ECK BSR SHC JK. Contributed reagents/materials/analysis tools: SF Y. Yen MMZ MO RC BSR SHC JK. Wrote the paper: SF TKD BZ ECK JK."} +{"text": "PLoS Genet 10(1): e1004100. doi:10.1371/journal.pgen.1004100The second author\u2019s name is spelled incorrectly. The correct name is: Oleg Evgrafov. The correct citation is: Staab TA, Evgrafov O, Knowles JA, Sieburth D (2014) Regulation of Synaptic"} +{"text": "The third author's name is misspelled. The correct spelling is: Brian P. Kavanagh. The correct citation is: Kroon AA, Wang J, Kavanagh BP, Huang Z, Kuliszewski M, et al. (2011) Prolonged Mechanical Ventilation Induces Cell Cycle Arrest in Newborn Rat Lung. PLoS ONE 6(2): e16910. doi:10.1371/journal.pone.0016910"} +{"text": "There were numerous errors throughout the article.The fifth author's name is misspelled. The correct name is: Bj\u00f6rn Brembs.http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0041642Reference 6 is incorrect. The correct reference is: Gomez-Marin A, Partoune N, Stephens GJ, Louis M (2012) Automated Tracking of Animal Posture and Movement during Exploration and Sensory Orientation Behaviors. PLoS ONE 7(8): e41642. doi:10.1371/journal.pone.0041642 Figure 1 is incorrect. The correct figure can be viewed here:"} +{"text": "The first author's name was misspelled. The correct name is: Jia-Qian Jiang. The correct citation is: Jiang J-Q, Zhou Z (2013) Removal of Pharmaceutical Residues by Ferrate(VI). PLoS ONE 8(2): e55729. doi:10.1371/journal.pone.0055729"} +{"text": "In Table 2, the table's symbol legends at the bottom are misaligned. Please see the corrected Table 2 here:http://www.plosntd.org/corrections/pntd.0001822.t002.cn.tif"} +{"text": "The cations are occupationally disordered and are located in the interlayer space. For both types of cations, distorted coordination polyhedra are observed.The title compound, potassium strontium trialuminium tetra\u00adarsenate, was prepared by solid-state reaction. The structure consists of AlO DOI: 10.1107/S1600536812014304/br2195Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S1600536812014304/br2195Isup3.cmlSupplementary material file. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "From thSources: McElhinny T. A mammalian lexicon. [cited 2005 Oct 13]. Available from http://www.msu.edu/~mcelhinn/zoology/mammalwords.htm; Webster's Third New International Dictionary (unabridged). Springfield (MA), 1993; and wikipedia.org."} +{"text": "This paper was published with an incorrect doi. The correct doi is: 10.1099/ijs.0.016451-0."} +{"text": "The Author Contributions are incorrect.They should be: Conceived the experiments: TBY GL. Designed the experiments: TB TBY GL. Performed the experiments: TB GL TBY. Analyzed the data: TB TBY GL YM. Wrote the paper: TBY TB ES."} +{"text": "The hydrogen-bond lengths determined from the structure refinement agree well with Raman spectroscopic data.The crystal structure of kovdorskite, ideally Mg al. 1980. Dokl. A DOI: 10.1107/S1600536812000256/wm2577Isup2.hklStructure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF reportAdditional supplementary materials:"} +{"text": "There were errors in the Author Contributions. The correct contributions are:Conceived and designed the experiments: BK DN PG JG EP SS JS.Performed the experiments: EP SS JS ML CCV MJ GG VN.Analyzed the data: BK DN ST PG JG EP CCV SS JS ML MJ GG.Contributed reagents/materials/analysis tools: SS EP NB HL.Wrote the paper: BK DN PG."} +{"text": "AbstractDistephanus for two species from Madagascar.New combinations are provided in Distephanus Cass. a number of combinations have already been made H. Rob., comb. nov. Basionym: Vernonia ambongensis Humbert, Bull. Soc. Bot. France 87: 346. 1940.urn:lsid:ipni.org:names:77122348-1Distephanus bara(Humbert) H. Rob., comb. nov. Basionym: Vernonia bara Humbert, Notul. Syst. (Paris) 8: 9\u201310. 1939.urn:lsid:ipni.org:names:77122349-1"} +{"text": "There is a typographical error in the title. The correct title is: Difference In Adaptive Dispersal Ability Can Promote Species Coexistence in Fluctuating Environments. The correct citation is: Lin W-T, Hsieh C-h, Miki T (2013) Difference In Adaptive Dispersal Ability Can Promote Species Coexistence in Fluctuating Environments. PLoS ONE 8(2): e55218. doi:10.1371/journal.pone.0055218"} +{"text": "Pseudoalteromonas arabiensis sp. nov. is erroneously cited as Pseudoalteromonas arabianensis sp. nov. in the title of the species description (page 1807).In this paper (doi:10.1099/ijs.0.043604-0), doi:10.1099/ijs.0.055459-0"} +{"text": "Int. J. Mol. Sci. 2011, 12, 2325\u20132335.Zhong Ye; Darya O. Mishchuk; Natasha S. Stephens and Carolyn M. Slupsky. Dextran Sulfate Sodium Inhibits Alanine Synthesis in Caco-2 Cells. Shu-Kun Linlin@mdpi.comMDPI AG, Postfach, CH-4005 Basel, Switzerland; E-Mail: Received: 8 January 2012 / Published: 9 February 2012It has been brought to our attention by the corresponding author that the results presented this article are in e"} +{"text": "Several errors are present in Figures 2 and 3. Please find the corrected versions of the figures here:http://plosone.org/corrections/pone.0060157.g002.cn.tifFigure 2: http://plosone.org/corrections/pone.0060157.g003.cn.tifFigure 3:"} +{"text": "The second author's name was spelled incorrectly. The correct name is: Michelle J. Sun.The correct citation is: Buskohl PR, Sun MJ, Thompson RP, Butcher JT (2012) Serotonin Potentiates Transforming Growth Factor-beta3 Induced Biomechanical Remodeling in Avian Embryonic Atrioventricular Valves. PLoS ONE 7(8): e42527. doi:10.1371/journal.pone.0042527"} +{"text": "Corresponding authorship was incorrectly omitted for two authors. The following are corresponding authors on this article: Hiroko Bannai (hbannai@brain.riken.jp), Antoine Triller (triller@biologie.ens.fr), and Katsuhiko Mikoshiba (mikosiba@brain.riken.jp)."} +{"text": "Due to errors introduced during the production process, Figure 1 and Figure 2 are incomplete. The full, correct figures can be found here:http://plosone.org/corrections/pone.0033023.g001.cn.tifFigure 1: http://plosone.org/corrections/pone.0033023.g002.cn.tifFigure 2:"} +{"text": "The authors note thahttp://www.biomedcentral.com/1741-7015/11/43.This is a Correction article onThe pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1741-7015/11/193/prepub"} +{"text": "Graphics 1 and 2 were omitted from the article. They are available below:http://www.plosone.org/corrections/pone.0071743.g001.1.cn.tifGraphic 1: http://www.plosone.org/corrections/pone.0071743.g002.1.cn.tifGraphic 2:"} +{"text": "Current Cardiology Reviews. Their efforts have contributed greatly tothe high quality and continuous growth of the journal. Given below is the list of reviewers who reviewed articles for the Journalduring 2013.Bentham Science Publishers would like to thank and appreciate the co-operation from all reviewers for their constructivecomments and feedback on the manuscripts submitted to S. Apostolakis (UK)K. Asrress (UK)A. Banerjee (UK)A. Baranchuk (Canada)H. Baumgartner (Germany)H. Baumgartner (Germany)A. Bobik R. Cemin G. J. Crystal (USA)P. M. Eckman (USA)M. Endoh (Japan)C. Foerch (Germany)S. Fujii (Japan)S. Fynn (UK)K. Guha (UK)K. Kallenbach (Germany)L .W. Klein (USA)L. H. Kuller (USA)J. L. Mehta (USA)P. Meier (UK)L. C. Naz\u00c1rio Scala (Brazil)P. Pagliaro M. A. Portman (USA)F. Recchia (USA)R. Sankaranarayanan (UK)R. J. Selvaraj (India)C. Shanahan (UK)E. Shantsila (UK)N. Turck (Switzerland)"} +{"text": "Abstractth century explorer and ornithologist Prince Maximilian of Wied-Neuwied form one of the foundation collections of the American Museum of Natural History in New York. However, parts of his collection remained in Germany and came to the Museum Wiesbaden. Since Wied described numerous new species without designating types, some of these specimens might be type material. Here we present a catalog of the 30 Wiesbaden specimens associated with him and discuss their potential type status. We conclude that 17 individuals in 11 species are potential type specimens that should be considered in future taxonomic work.Bird specimens collected by 19 In 1870 part of the Wied collection comprising 4,000 birds, 600 mammals and 2,000 fish and reptiles was purchased by the American Museum of Natural History in New York (AMNH) and constitutes the cornerstone of its scientific collection owns several specimens that either originate from the collection of Prince Maximilian of Wied-Neuwied or were described by him as new species. In the early 19llection .Besides mammals, reptiles, and amphibians, Wied described 160 species and subspecies of birds. The names of more than 50 of these taxa are still valid today . PotentiLike at the AMNH, in Wiesbaden as well many of the originally mounted specimens were later dismounted and added to the study skin collection. Most of the series of the main Wied collection at the AMNH still bear their original labels, but not every individual specimen has such a label by Wied see . It appeWied\u2019s work on birds is significant not only because of the huge number of species and forms described for the first time, but it also gives information on distribution and biology of numerous animals. Even today records on the biology of many organisms are completely lacking. As PageBreakHandbook of the Birds of the World Arctic Tern \u2013 K\u00fcstenseeschwalbeInv. nr. 2209: 1 ad., breeding plumage, mounted specimenLabels: a) [*] 2209. Cat. Birds Br. Mus. XXV, 62. Sterna macrura Naum. S\u00fcdamerika Fr\u00fchjahr 1846 G.: Prinz Max v. Wied; b) [**] 2209 Rchw. 1, 116. Sterna macrura Naum. K\u00fcsten-Seeschwalbe (Sommerkleid) S\u00fcd-Amerika G.: Prinz Max v. Wied.Anser albifrons albifrons 2. (Anseriformes \u2013 Anatidae)Greater White-fronted Goose \u2013 Bl\u00e4\u00dfgansInv. nr. 2504: 1 fem. juv., mounted specimenLabels: a) Anser albifrons, Bechst. Bl\u00e4\u00dfgans \u2640 N.Europa; b) [*] 2504. Cat. Birds Brit. Mus. 27 pag. 92. Anser albifrons (Scop.) \u2640 N.Europa Novbr. 1847 G.: Prinz Max v. Wied; c) [**] 2504. Anser albifrons (Scop.) Wei\u00dfstirngans, Bl\u00e4\u00dfgans \u2640 juv. Europa XI/1847 G.: Prinz Max v. WiedMeleagris gallopavo osceola Scott, 18903. Florida Wild Turkey \u2013 Florida-TruthuhnInv. nr. 1853: 1 fem. ad., mounted specimenLabels: a) Meleagris Gallopavo Nord-Amerika; b) [*] 1853 Cat. B. Br. Mus. XXII, 389/90 Meleagris americana Bartr. subsp. osceola Scott \u2640 Nord-Amerika 1835. G.: Prinz Max v. Wied; c) [**] 1853 Rchw. 1, 304. Meleagris americana Bartr. subsp. osceola Scott Wildes Truthuhn Nord-Amerika S.G.: S. H. Prinz M. v. WiedPageBreakPageBreakAulacorhynchus prasinus atrogularis 4. (Piciformes \u2013 Ramphastidae)Emerald Toucanet \u2013 LaucharassariInv. nr. 848: 1 ad., mounted specimenLabels: a) Pteroglossus atrogularis Gould. Amerika Von Prinz Max erkauft(?) [purchased from Prince Max]; b) [*] 848. Br. C. B. XIX p. 160 Aulacorhamphus atrogularis Sturm S.Amerika S. Prinz Max v. Wied; c) [**] 848 Rchw. 2, 37. Aulacorhynchus atrogularis (Sturm.) Schwarzkehliger Arassari S\u00fcdamerika.; on pedestal: 1835 ans Museum [to the museum in 1835]Colaptes auratus auratus -group5. (Piciformes \u2013 Picidae)Yellow-shafted Flicker \u2013 GoldspechtInv. nr. 1002: 1 fem. ad., study skinLabels: a) Picus auratus Nord-Amerika; b) [*] 1002 Cat. Birds Brit. Mus. 18.p.12. Colaptes auratus (L.) \u2640 Nord-Amerika; catalog: Prinz Max von Wied, 1835Melanerpes erythrocephalus 6. (Piciformes \u2013 Picidae)Red-headed Woodpecker \u2013 RotkopfspechtInv. nr. 1027: 1 ad., study skinLabel: [***] Kat.Nr. 1027 Br.C.B. XVIII 145 Melanerpes erythrocephalu (L) Rotkopfspecht Nord-Amerika; database: erworben 1835 von Prinz Max v. Wied [purchased 1835 from Prince Max of Wied]Sporophila caerulescens 7. (Passeriformes \u2013 Thraupidae)Double-collared Seedeater \u2013 Schmuckpf\u00e4ffchenInv. nr. 5238: 1 male ad., study skinLabel: [***] Kat.Nr. 5238. Sporophila ornata (Licht.) \u2642 Brasilien leg. Prinz Max v. Wied; reverse: Sporophila (-)caerulescens (Vieill.) Schmuckpf\u00e4ffchenTouit melanonota 1. (Psittaciformes \u2013 Psittacidae)Brown-backed Parrotlet \u2013 Braunr\u00fcckenpapageiInv. nr. 748: 1 ad., mounted specimenSyn.: Psittacus melanonotus Wied, 1820: 275; Urochroma melanota Labels: a) Psittacus melanotus Brasilien; b) [*] 748 Br.C.B. XX. p. 352. Urochroma wiedi Allen Brasilien.; c) [**] 748 Rchw. 1, 484. Urochroma melanota (Lcht.) Wied Schwarzr\u00fcckiger Zwergpapagei Brasilien.; catalog: Prinz Max v. Wied, 1835Remarks: According to the original description, Wied saw several animals and noted that this new species was displayed at the Berlin museum under the name PageBreakPageBreakPsittacus melanonotus. Psittacus, Wied\u2019s name was preoccupied by Psittacus melanotus Shaw, 1804. Allen provided a replacement name, Urochroma wiedi. Wied\u2019s form is now placed in the genus Touit 2. (Strigiformes \u2013 Strigidae)Least Pygmy-owl \u2013 KleinstzwergkauzInv. nr. 202: 1 ad., study skinSyn.: Strix minutissima Wied, 1830: 242Label: [*] 202 Glaucidium (Glaucidium) minutissimum (Wied) Zwergkauz BrasilienRemarks: According to the original description, Wied examined males and females, but does not give any information about the quantities or the whereabouts. Strix minutissima at the AMNH with the inventory numbers 6345 and 6345 bis . The MWNH specimen might belong to the original type series, although there is no proof.Spizaetus tyrannus 3. Black Hawk-eagle \u2013 TyrannenadlerI. Inv. nr. 2989: 1 male ad., mounted specimenII. Inv. nr. 2990: 1 juv., study skinSyn.: Falco tyrannus Wied, 1820: 360Labels: I. a) No. 109 Falco Tyrannus Pr. Max Temm. Pl. col. 73. Seite 61 Cuv. Seite 384. Iris gelb.; b) [*] 2989. Brit. Cat. I. 264 Spizaetus tyrannus Wied. \u2642 Surinam; c) [**] 2989 Rchw. 1, 386. Spizaetus tyrannus (Wied.) Tyrann-Adler \u2642 Surinam.; II. [***] Kat.Nr. 2990 R. I. 386 Spizaetus tyrannus (Wied) Tyrann-Adler SurinamRemarks: According to the original description, Wied had at least one male at his hands. The AMNH has a specimen (inv. nr. 6381) that Conopias trivirgatus 4. (Passeriformes \u2013 Tyrannidae)Three-striped Flycatcher \u2013 Olivbrust-MaskentyrannInv. nr. 5313: 1 ad., study skinSyn.: Muscicapa trivirgata Wied, 1831: 871Labels: a) Muscicapa trivirgata unserer(?) Beitr\u00e4ge; b) Muscicapa trivirgata M. v. Wied, Brasilien; c) [*] 5313. Cat. Birds Brit. Mus. Conopias (Conopias) trivirgata (Wied) Subsp. Dreistreifentyrann BrasilienPageBreakPageBreakRemarks: According to the original description, Wied examined one female from Bahia. Femina\u201d was not written in Wied\u2019s catalog, but AMNH 4926 is sexed as a female. Since Wied himself only mentions one female, the specimen at the AMNH probably is the holotype.Lipaugus vociferans 5. (Passeriformes \u2013 Cotingidae)Screaming Piha \u2013 Tiefland-GraupihaI. Inv. nr. 150: 1 ad., mounted specimenII. Inv. nr. 3989: 1 fem. ad., study skinSyn.: Muscicapa vociferans Wied, 1820: 242Labels: I. a) [*] 150 Cat. Birds Brit. Mus. 14 p.352. Lathria cinerea (Vieill.) Surinam; b) [**] 150 Rchw. 2, 191. Lipaugus cinereus (Vieill.) Surinam.; II. [*] 3989. R. II. 191. Lipaugus cinereus (Vieill.) Brasilien G.: Geschw. Brambeer.; reverse: CotingidaeRemarks: In the original description Wied does not state the quantity of examined specimens. According to him, this new species was displayed at the Berlin museum under the name Muscicapa ampelina. Todirostrum poliocephalum 6. (Passeriformes \u2013 Tyrannidae)Grey-headed Tody-flycatcher \u2013 Gelbz\u00fcgel-TodityrannI. Inv. nr. 120 a: 1 ad., study skinII. Inv. nr. 120 b: 1 ad., study skinIII. Inv. nr. 120 c: 1 ad., study skinSyn.: Todus poliocephalus Wied, 1831: 964Lab els: I. a) Todus poliocephalus, Pr. Max Grauk\u00f6pfiger Plattschnabel Brasilien; b) [*] 120 3/a Cat. Birds Brit. Mus. 14 p. 71. Todirostrum poliocephalum (Wied) Brasilien.; II. a) Todus poliocephalus Max v. Wied. Grauk\u00f6pfiger Plattschnabel Brasilien; b) [*] 120 3/b Cat. Birds Brit. Mus. 14. 71 Todirostrum poliocephalum (Wied.) Brasilien; III. a) Todus policephalus, Max v. [abgeschnitten] Grauk\u00f6pfiger Plattschnabel Brasilien; b) [*] 120 3/c Cat. Birds Brit. Mus. 14 p. 71 Todirostrum poliocephalum (Wied) Brasilien.; reverse of b) : TyrannidaeRemarks: In the original description Wied examined males and females, but did not note the quantities and whereabouts. PageBreakConopophaga lineata 7. (Passeriformes \u2013 Conopophagidae)Rufous Gnateater \u2013 Rotkehl-M\u00fcckenfresserI. Inv. nr. 106 a: 1 ad., study skinII. Inv. nr. 106 b: 1 ad., study skinIII. Inv. nr. 107: 1 ad., study skinSyn.: Myiagrus lineatus Wied, 1831: 1046Labels: I. [*] 106a. Cat. Birds Brit. Mus. 15. p. 333 Conopophaga lineata (Wied) Brasilien; II. [*] 106b. Cat. Birds Brit. Mus. 15 p. 333. Conopophaga lineata (Wied.) Brasilien; III. [*] 107 Cat. Birds Brit. Mus. 15 p. 333. Conopophaga lineata (Wied.) BrasilienRemarks: In the original description Wied mentions only one individual. The AMNH has one female holotype (Nr. 6777) listed by Myrmeciza ruficauda 8. (Passeriformes \u2013 Thamnophilidae)Scalloped Antbird \u2013 N\u00f6rdlicher SchuppenameisenvogelInv. nr. 121: 1 ad., mounted specimenSyn.: Myiothera ruficauda Wied, 1831: 1060Labels: a) Formicivora loricata \u2640 Swains. Bahia; b) Formicivora \u2642. Bahia. 3. Gust. Schneider, Basel.; c) [*] 121. Cat. Birds Brit. Mus. 15 p. 281. Myrmeciza ruficauda Wied Bahia S.: G. Schneider, Basel.; d) [**] 121 Rchw. 2, 231. Myrmeciza ruficauda Wied. Bahia.Remarks: In the original description Wied examined males and females, but does not specify the quantities and whereabouts. There are four syntypes at the AMNH: males nr. 5388 and 6829, juvenile male nr. 5386 and female nr. 5385 9. (Passeriformes \u2013 Corvidae)White-naped Jay \u2013 Wei\u00dfnacken-BlaurabeInv. nr. 609: 1 ad., mounted specimenSyn.: Corvus cyanopogon Wied, 1821: 137Labels: a) Corvus cyanopogon Brasilien; later altered: Cyanocorax Corvus cyanopogon (Max Neuwied) III, 123.; b) [*] 609 Br. C. B. III p. 123 Cyanocorax cyanopogon (Neuwied) Brasilien.Remarks: In the original description Wied had several individuals at hand. Tangara velia cyanomelas Wied, 183010. (Passeriformes \u2013 Thraupidae)Silvery-breasted Tanager \u2013 Rotbauchtangare; group of 3 individualsI. Inv. nr. 3792: 1 male ad., mounted specimenII. Inv. nr. 3793: 1 fem. ad., mounted specimenIII. Inv. nr. 3794: 1 fem. ad., mounted specimenPageBreakSyn.: Tangara cyanomelas Wied, 1830: 453Labels: a) Tanagrella velia \u2642\u2640? Gmel Bahia; b) [*] 3792/94 Brit.Cat.11 p. 88. Tanagrella cyanomelaena (Wied.) \u2642\u2640\u2640 Bahia; c) [**] 3792/94 Rchw. 2, 436. Calospiza cyanomlaena (Wied.) 1 \u2642, 2 \u2640 Bahia, Brasilien.Remarks: In the original description Wied mentions several males, while the female was unknown to him. According to Coryphospingus pileatus 11. (Passeriformes \u2013 Thraupidae)Pileated Finch \u2013 Graur\u00fcckenkronfinkInv. nr. 5102: 1 male ad., study skinSyn.: Fringilla pileata Wied, 1821: 160Label:[*] 5102. Cat. Birds Brit. Mus. Tanagra cristatella Spix. \u2642 Brasilien; reverse: Tanagridae = ThraupidaeCoryphospingus pileatus (Wied) Subsp. Graur\u00fccken-KronfinkRemarks: In the original description Wied described the male without giving details on quantity or whereabouts of the specimens. According to Schistochlamys ruficapillus capistratus 12. (Passeriformes \u2013 Thraupidae)Cinnamon Tanager \u2013 GimpeltangareInv. nr. 5095: 1 ad., study skinSyn.: Tanagra capistrata Wied, 1821: 179Label: [*] 5095. Brit.Cat.11 p. 301. Schistochlamys capistratus (Max) Brasilien.; reverse: Tanagridae = ThraupidaeRemarks: In the original description, and also in 1831 (p. 500), Wied examined males and females, but did not note specifics on quantities and whereabouts. According to Thraupis palmarum palmarum 13. (Passeriformes \u2013 Thraupidae)Palm Tanager \u2013 PalmentangareI. Inv. nr. 43: 1 ad., mounted specimenPageBreakII. Inv. nr. 5096: 1 ad., study skinSyn.: Tanagra palmarum Wied, 1821: 76Labels: I. a) Tanagra palmarum Mexiko; b) [*] 43 Brit.Cat. 11 p. 159. Tanagra palmarum Max. Brasilien. Mexiko auf alt/Schauetik.; c) [**] 43 Rchw. 2, 435. Tanagra palmarum Max. Brasilien.; II. [*] 5096. Brit.Cat.11 p.159. Tanagra palmarum Max. Brasilien.Remarks: In the original description Wied refers to both sexes, but does not specify the quantity and whereabouts of the examined specimens. Oryzoborus maximiliani Cabanis, 185114. (Passeriformes \u2013 Thraupidae)Great-billed Seed-finch \u2013 Dickschnabel-ReisknackerInv. nr. 5235: 1 fem. ad., study skinSyn.: Oryzoborus crassirostris Wied, 1830: 564, preocc. Oryzoborus crassirostris Labels: a) Fringilla crassirostris, Max v. Wied Pyrrhula crassirost. \u2640 Brasilien; b)[*] 5235. Brit.Cat.12 p. 78 Oryzoborus maximiliani Cab. \u2640 Brasilien.Remarks: Unfortunately we could not obtain the original description. In conclusion, it can be assumed that 17 individuals in 11 species of birds in the Wiesbaden collection are potential type specimens that should be considered in future taxonomic work. Close examination, comparison with other material, or even genetic tests will be necessary to make a final decision on the specimens\u2019 type status."} +{"text": "There were multiple errors in the legends for Figures 2, 3 and 4 in the online version of the article, and in Figure 2 of the PDF version of the article. The correct legends can be viewed below.http://www.plosone.org/corrections/pone.0075177.002.cn.tiffFigure 2: http://www.plosone.org/corrections/pone.0075177.003.cn.tiffFigure 3: http://www.plosone.org/corrections/pone.0075177.004.cn.tiffFigure 4:"} +{"text": "The name of the first author was given incorrectly. The correct name is: Jacob Keller. The correct citation is: Keller J, Homma K, Dallos P (2013) Pixels as ROIs (PAR): A Less-Biased and Statistically Powerful Approach for Gleaning Functional Information from Image Stacks. PLoS ONE 8(7): e69047. doi:10.1371/journal.pone.0069047."} +{"text": "The image for Figure 1 is incorrect. Please view the correct image here:http://www.plosntd.org/corrections/pntd.0001902.g001.cn.tif"} +{"text": "AbstractFirensia Scop. was based on Cordia flavescens Aubl., a species described and illustrated from a mixed collection that Scopoli never transferred to Firensia. The genus included three additional species formally named by Rafinesque. Currently the four species are placed in three different families and none retained the epithet accepted by Scopoli or given by Rafinesque for reason of priority. A lectotype is designated for Cordia flavescens that places Firensia in the synonymy of Ocotea (Lauraceae). Boraginaceae for the Flora of the Guianas, I got intrigued by the history of Firensia Scop. When Firensia, he included only Cordia flavescens Aubl. that is therefore the type of the genus. Contrary to what is stated by http://www.ipni.org/index.html; accessed 23.01.2013), in that work Cordia flavescens was not transferred to Firensia. Firensia as one PageBreakof his \u201cspecies naturalis\u201d in a work where the nomenclature was uninominal and that is rejected as a source of names by the International Code of Botanical Nomenclature \u201d. When the original description was associated with several collections of equal status or syntypes, a type collection and a lectotype were selected and the text says: \u201cLectotype...: ... \u201d. In both cases the information on the date of lectotypification is given next to the word lectotype.Cordia flavescens Aubl. was described and illustrated based on a mixture of fruiting branches of Ocotea commutata Nees (LAURACEAE) and flowers belonging in Cordia. Specimens by Aublet at BM and S represent only the Ocotea element does not have priority over Ocotea Aubl. (1775), and the epithet flavescens cannot be transferred to Ocotea as the name Ocotea flavescens Rusby (1920) applies to a different species from Colombia. Aublet\u2019s name becomes a synonym of Ocotea commutata (Nees) Mez.Nomenclature stability is satisfied: PageBreakOcotea Aubl., Hist. Pl. Guiane 2: 780; 4: t. 310. 1775.Ocotea guianensis Aubl.Type: Firensia Scop., Intr. Hist. Nat. 157. 1777.Cordia flavescens Aubl.Type: Firensia have been named or identified as species of Cordia L., one by Firensia could be kept for reason of priority. The type species, Cordia flavescens, and the three species of Firensia currently belong in three different genera and families.The species once placed in 1.Aubl., LauraceaeOcotea commutataOreodaphne commutata Nees (Nees) Mez, Jahrb. K\u00f6nigl. Bot. Gart. Berlin 5: 327. 1889. Type: Based on Cordia flavescensOcotea flavescens Rusby 1920. Type: French Guiana. J.B. Aublet s.n. [specimens without the Cordia flowers] Aubl., Hist. Pl. Guiane 1: 226; 3: t. 89. 1775; not Cordia sarmentosaCordia flavescens Aubl. Lam., Tab. Encycl. M\u00e9thod. 1: 422. 1791; illegitimate renaming. Type: Based on Oreodaphne commutataJ. Martin s.n. Kuntze, Revis. Gen. Pl. 2: 977. 1891. Type: Based on Gerascanthus flavescensCordia flavescens Aubl. (Aubl.) Borhidi, Acta Bot. Hung. 34(3\u20134): 400. 1988. Type: Based on Oreodaphne commutata can be seen at http://ww2.bgbm.org/herbarium/view_large.cfm?SpecimenPK=47688&idThumb=253360&SpecimenSequenz=1&loan=0 , and the Paris isotypes at http://coldb.mnhn.fr/colweb/request.do?requestaction=exec; the lectotype of Cordia flavescens at http://plants.jstor.org/specimen/bm000993950 , and its Stockholm isotype at http://plants.jstor.org/specimen/s04-2350?history=true .The holotype of PageBreak2.Raf., CordiaceaeCordia collococcaP. Browne s.n. L., Sp. Pl. ed. 2, 1: 274. 1762. Lectotype Kuntze, Revis. Gen. Pl. 2: 438. 1891. Type: Based on Gerascanthus collococcusCordia collococca L. (L.) Borhidi, Acta Bot. Hung. 34(3\u20134): 399. 1988. Type: Based on Cordia collococca can be seen at .The hololectotype of For other synonyms, see 3.(Willd.) Raf., CordiaceaeCordia nodosa Lam., Tab. Encycl. 1: 422. 1792. Type: Cordia collococcaCordia nodosa Lam.) sensu J.B. Aublet, Hist. Pl. Guiane 1: 219; & 3: pl. 86. 1775; non L. 1757. : 1076. 1798. Type: Based on Firensia hirsutaCordia nodosa Lam. (Willd.) Raf., Sylva Tellur. 40. 1838. Type: Based on Lithocardium nodosumCordia nodosa Lam. (Lam.) Kuntze, Rev. Gen. 2: 977. 1891. Type: Based on Cordia nodosa and Cordia hirsuta. In fact it is unlikely that Sometimes the collection from French Guiana\u2014J.B. Aublet s.n. \u2014is cited as the type of http://plants.jstor.org/specimen/bm000906214Aublet s.n. can be seen at Cordia hirsuta Fresen. 1857 is different from Cordia hirsuta Willd. 1798.For other synonyms, see 4.Raf., CombretaceaeBuchenavia tetraphyllaCordia tetraphylla Aubl. (Aubl.) R.A. Howard, J. Arnold Arbor. 64(2): 266. 1983. Type: Based on PageBreakCordia tetraphylla Aubl., Hist. Pl. Guiane 1: 224; 3: t. 88. 1775. Lectotype Kuntze, Rev. Gen. 2: 977. 1891. Type: Based on Gerascanthus tetraphyllusCordia tetraphylla Aubl. (Aubl.) Borhidi, Acta Bot. Hung. 34(3\u20134): 402. 1988. Type: Based on For other synonyms, see Ocotea commutata Nees ex Meisn., Prodr. (DC.) 15(1): 120. 1864, invalid: pro syn. Oreodaphne commutata Nees (= Ocotea commutata (Nees) Mez 1889).Cordia echitioides Lam. ex D. Dietr., Syn. Pl. 1: 612. 1839; invalid: pro syn. Cordia flavescens Aubl. (= Ocotea commutata (Nees) Mez)."} +{"text": "The eighth author's name was spelled incorrectly. The correct name is: Montserrat Carmona.The title of Reference 16 is incorrect. The corrected Reference 16 is:http://www.isciii.es/ISCIII/es/contenidos/fd-servicios-cientifico-tecnicos/fd-vigilancias-alertas/M_10_Cap_cau.xls. Accesssed 17 September 2013.16. Instituto de Salud Carlos III website. (2010) . In the footnote of Table 3, the text for a. references Figure 4 erroneously. The correct text should read: \"a. Correspond to overall values in Figure 3.\""} +{"text": "Figures 2B through 2E do not appear.http://www.plosone.org/corrections/pone.0066792.g002b.cn.tifPlease view Figure 2B here: http://www.plosone.org/corrections/pone.0066792.g002c.cn.tifPlease view Figure 2C here: http://www.plosone.org/corrections/pone.0066792.g002d.cn.tifPlease view Figure 2D here: http://www.plosone.org/corrections/pone.0066792.g002e.cn.tifPlease view Figure 2E here:"} +{"text": "A typicl Figure . Insteadt Figure . Spatialparisons . These r"} +{"text": "Supplementary Videos S1 and S2 and Supplementary Tables S1 through S4 were incorrectly omitted from the Supporting Information. Please view the Supplementary Videos and Tables at the following links: http://plosone.org/corrections/pone.0079913.s001.cn.mp4 Video S1: http://plosone.org/corrections/pone.0079913.s002.cn.mp4 Video S2: http://plosone.org/corrections/pone.0079913.s003.cn.tif Table S1: http://plosone.org/corrections/pone.0079913.s004.cn.tif Table S2: http://plosone.org/corrections/pone.0079913.s005.cn.tif Table S3: http://plosone.org/corrections/pone.0079913.s006.cn.tifTable S4:"} +{"text": "The Table 1 that was presented is incorrect. The correct version is available below.http://plosone.org/corrections/pone.0050295.t001.cn.tifTable 1:"} +{"text": "The last author's name was misspelled. The correct name is: Glenn J. Tattersall. The correct citation is: Greenberg R, Cadena V, Danner RM, Tattersall GJ (2012) Heat Loss May Explain Bill Size Differences between Birds Occupying Different Habitats. PLoS ONE 7(7): e40933. doi:10.1371/journal.pone.0040933"} +{"text": "There were numerous errors throughout the article. http://www.plosone.org/corrections/pone.0049593.cn.tifThe corrections can be viewed here:"} +{"text": "The middle initial of the second author is incorrect. The correct spelling is: Peter N. Ruygrok.The correct citation is: Crossman DJ, Ruygrok PN, Soeller C, Cannell MB (2011) Changes in the Organization of Excitation-Contraction Coupling Structures in Failing Human Heart. PLoS ONE 6(3): e17901. doi:10.1371/journal.pone.0017901The correct author contributions are: Conceived and designed the experiments: MBC DJC PNR. Performed the experiments: DJC. Analyzed the data: MBC DJC CS. Contributed reagents/materials/analysis tools: MBC CS PNR. Wrote the paper: DJC MBC."} +{"text": "Mycobacterium tuberculosis Complex Pathogen, M. mungi contained several errors related to scoring of mycobacterial interspersed repetitive unit\u2013variable number tandem repeats of selected isolates. The article has been corrected online (http://wwwnc.cdc.gov/eid/article/16/8/10-0314_article.htm). Table 2 in the article Novel"} +{"text": "The data in doi:10.1099/mic.0.X00007-0"} +{"text": "There was an error in the second author's name. Peter J. Carolan is correct. The correct citation is: Chen CA, Carolan PJ, Annes JP (2012) In Vivo Screening for Secreted Proteins That Modulate Glucose Handling Identifies Interleukin-6 Family Members as Potent Hypoglycemic Agents. PLoS ONE 7(9): e44600. doi:10.1371/journal.pone.0044600"} +{"text": "In \u201cOsler and the Infected Letter,\u201d by Charles T. Ambrose, an error occurred. Yellow fever swept through Philadelphia in 1793.http://wwwnc.cdc.gov/eid/article/11/5/04-0616_article.htm.The corrected article appears online at We regret any confusion these errors may have caused."} +{"text": "The fifth author's name is spelled incorrectly. The correct name is: Aparna Wagle Shukla.The correct citation is: Hass CJ, Malczak P, Nocera J, Stegem\u00f6ller EL, Wagle Shukla A, et al. (2012) Quantitative Normative Gait Data in a Large Cohort of Ambulatory Persons with Parkinson\u2019s Disease. PLoS ONE 7(8): e42337. doi:10.1371/journal.pone.0042337"} +{"text": "Due to errors introduced during the production process, some of the text has been reproduced incorrectly.http://www.plosone.org/corrections/pone.0052603.001.cn.tif In the Results section, in the sub-section \"Incorporation of Gag-GFP into Yeast VLPs does not Require ESCRT Function\", errors were introduced into the text. The correct text can be viewed here: http://www.plosone.org/corrections/pone.0052603.002.cn.tif In the Materials and Methods section, in the sub-section \"Strains\", errors were introduced into the text. The correct text can be viewed here: In the legends of Figure 1, Figure 4, and Figure 5, errors were introduced into the text. The correct legends can be found below.http://www.plosone.org/corrections/pone.0052603.003.cn.tif Figure 1: http://www.plosone.org/corrections/pone.0052603.004.cn.tif Figure 4: http://www.plosone.org/corrections/pone.0052603.005.cn.tif Figure 5:"} +{"text": "There was an error in the Author Contributions designations. The correct Author Contributions are: Conceived and designed the experiments: PF PT GK CP AHF. Performed the experiments: PF ML CP. Analyzed the data: PF ML GK AJF NR. Contributed reagents/materials/analysis tools: PF ML PM CP NR. Wrote the paper: PF PT GK BW ML PM. Provided data: PM AHF GK BW."} +{"text": "Recurrent/Persistent Pneumonia among Children in Upper Egypt. Mediterr J Hematol Infect Dis. 2013 Apr 18;5(1):e2013028. doi: 10.4084/MJHID.2013.028. Print 2013. PubMed PMID: 23667726;PubMed Central PMCID: PMC3647710.\" has been removed from Mediterr J Hematol Infect Dis because the paper was previously published in another journal.The article deposited in PUBMED as"} +{"text": "Sensors [http://www.mdpi.com/1424-8220/15/4/9277.The authors wish to update the Acknowledgments in their paper published in Sensors , doi:10."} +{"text": "The third and fourth author's names are spelled incorrectly. The correct names are: Gert S. Faber and N. Peter Reeves. The correct citation is: Cofr\u00e9 Lizama LE, Pijnappels M, Faber GS, Reeves NP, Verschueren SM, et al. (2014) Age Effects on Mediolateral Balance Control. PLoS ONE 9(10): e110757. doi:10.1371/journal.pone.0110757"} +{"text": "Footnotes 6 and 7:http://fcon_1000.projects.nitrc.org/indi/abide/Should both refer to the ABIDE dataset: Footnote 8:http://neuro.debian.net.Should reference the article below, in addition to the NeuroDebian website: Front. Neuroinform. 6:22. doi: 10.3389/fninf.2012.00022. http://journal.frontiersin.org/article/10.3389/fninf.2012.00022/pdfHalchenko, Y. O., and Hanke, M. (2012). Open is not enough. Let's take the next step: an integrated, community-driven computing platform for neuroscience. The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Present: Due to a technical error during publication, duplicate videos were uploaded as Supplementary File 5 and Supplementary File 28. As a result, Figure 5E contains a reference to the duplicate files.Corrected: Correct Supplementary File 28 was uploaded. Updated figure 5E can be found below. The publisher apologizes for this oversight.21655-21674. doi: 10.18632/oncotarget.4130.Original Article: Oncotarget. 2015; 6:"} +{"text": "There are errors in the Author Contributions. The correct contributions are: Conceived and designed the experiments: AM FT AG CC LR. Performed the experiments: FT DL GB AM. Analyzed the data: AM FT AG CC LR. Contributed reagents/materials/analysis tools: FV. Wrote the paper: AM FT AG LR"} +{"text": "Jaroma A, Soininvaara T, Kr\u00f6ger H. Periprosthetic tibial bone mineral density changes after total knee arthroplasty. ACTA ORTHOP 2016; 87 (3): 268-273.http://dx.doi.org/10.3109/17453674.2016.1173982When the above article was first published online, co-author Tarja Soininvaara\u2019s surname was incorrectly spelt as Soinninvaara. This has now been corrected online and in the print edition."} +{"text": "Nucl. Acids Res. (2013) 41 (18): e175. doi: 10.1093/nar/gkt684The authors wish to correct the full name of Billie Jo Masek to Billie J. Masek."} +{"text": "Bulletin \u00e9pid\u00e9miologique hebdomadaire 24-25/201626 July 2016, Francehttp://invs.santepubliquefrance.fr/beh/2016/24-25/2016_24-25_3.htmlHealth Protection Report; 10(23)15 July 2016, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/541028/hpr2316.pdfHealth Protection Report; 10(24)22 July 2016, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/541032/hrp2416.pdfHealth Protection Report; 10(23)15 July 2016, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/541028/hpr2316.pdfHealth Protection Report; 10(22)08 July 2016, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/543433/hpr2216_crrctd2.pdfFolkehelseinstituttet website20 July 2016https://www.fhi.no/nyheter/2016/oppdaterte-rad-om-forebygging-av-zikavirus-infeksjon-etter-opphold-i-utbrud/Health Protection Report; 10(24)22 July 2016, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/541032/hrp2416.pdfEpidemiologisches Bulletin 29, 201625 July 2016, Germanyhttp://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2016/Ausgaben/29_16.pdf?__blob=publicationFileHealth Protection Report; 10(24)22 July 2016, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/541032/hrp2416.pdfFolkehelseinstituttet website07 July 2016https://www.fhi.no/nyheter/2016/mat-og-vannbarne-infeksjoner/Epi-Insight 2016;17(8)August 2016, Irelandhttp://ndsc.newsweaver.ie/epiinsight/xvz2wrpu8jr?a=1&p=50630336&t=17517774EPI-NEWS 27a+b, 20166 July 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2027%20-%202016.aspxEpidemiologisches Bulletin 30, 20161 August 2016, Germanyhttp://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2016/Ausgaben/30_16.pdf?__blob=publicationFileHealth Protection Report; 10(24)22 July 2016, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/541032/hrp2416.pdf"} +{"text": "The fourth author\u2019s name is spelled incorrectly. The correct name is David Johansen. The correct citation is: Songstad NT, Serrano MC, Sitras V, Johansen D, Ytrehus K, et al. (2014) Coronary Flow Reserve in Pregnant Rats with Increased Left Ventricular Afterload. PLoS ONE 9(7): e102147. doi:10.1371/journal.pone.0102147."} +{"text": "Tunga penetrans Antigens in Selected Epidemic Areas in Murang\u2019a County in Kenya. PLoS Negl Trop Dis 9(3): e0003517. doi:10.1371/journal.pntd.0003517.The third author\u2019s name is spelled incorrectly. The correct name is Joshua M. Mutiso. The correct citation is: Mwangi JN, Ozwara HS, Mutiso JM, Gicheru MM (2015) Characterization of"} +{"text": "AbstractIn order to contribute to the butterflies\u2019 biodiversity knowledge at Serra do Intendente State Park - Minas Gerais, a study based on collections using Van Someren-Rydon traps and active search was performed. In this study, a total of 395 butterflies were collected, of which 327 were identified to species or morphospecies. 263 specimens were collected by the traps and 64 were collected using entomological hand-nets; 43 genera and 60 species were collected and identified. Lepidoptera is comprised of butterflies and moths; it is one of the main orders of insects which has approximately 157,424 described species (Papilionoidea and Hesperioidea and are subdivided into six families: Hesperiidae (Hesperioidea) and Papilionidae, Pieridae, Lycaenidae, Riodionidae, and Nymphalidae (Papilionoidea) (The species . The but species . The occ species . The groonoidea) .These insects are characterized as holometabolous, terrestrial, and diurnal. They are plant material chewers in the larval stage and liquid suckers in adulthood . ButterfButterflies are important indicators of environmental quality, because they are diverse, can be easily viewed, captured, identified, and manipulated by researchers . They arLepidoptera biodiversity in Minas Gerais, is still scarce , Minas Gerais, Brazil.Study SiteThe study was conducted in the region of Serra do Espinha\u00e7o, more precisely, within the Serra do Intendente State Park Fig. and the The climate is mesothermal, characterized by mild, humid summers and dry, cold winters. The average annual rainfall is 1,600 mm. The annual mean temperature is 18, 7\u00baC . The preData CollectionThe collections began in April 2012 and were completed in February of 2013. During this period four collections (two in the rainy season and two in the dry season) were performed. Each collection was performed for five days. The study area was divided into two areas throughout the Peixe Tolo River basin and in each area, forty Van Someren-Rydon traps were distributed. Twenty traps were located on the right bank and other twenty on the left bank of the Peixe Tolo River . Genera and species were confirmed by Dr. Andr\u00e9 Freitas, from the Department of Animal Biology, Universidade Estadual de Campinas. Furthermore, a comparison with available identified species in the Lepidoptera collection in the Invertebrates Laboratory (PUC Minas) was performed.The collected material was mounted, identified and labeled in the PUC Minas Natural Sciences Museum entomological collection laboratory. The identification of the individuals was made using In this study 394 individuals were captured, and 327 were identified. Sixty-seven individuals were not identified to genus or species due to bad specimen conditions or incipient systematics.Nymphalidae, Pieridae, Hesperiidae, Lycaenidae, Papilionidae and Riodinidae. A total of 299 individuals belonging to the Nymphalidae, 15 from the Pieridae, four from the Hesperiidae, four from the Lycaenidae, three from the Papilionidae, and one species from the Riodinidae.The families represented in this study were: A total of 263 butterflies were collected in traps and 63 using entomological hand-nets. The collections gathered specimens belonging to 43 genera and 60 species Table . During http://eol.org/pages/4090956/overviewhttp://www.butterfliesofamerica.com/L/adelotypa_malca.htmhttp://eol.org/pages/168926/overviewhttp://butterfliesofamerica.com/L/archaeoprepona_d_demophoon_types.htmhttp://eol.org/pages/181967/overviewhttp://eol.org/pages/159066/overviewhttp://butterfliesofamerica.com/anartia_a_amathea_types.htmhttp://eol.org/pages/172859/overviewhttp://www.butterfliesofamerica.com/L/ascia_m_monuste_types.htmhttp://eol.org/pages/11555451/overviewhttp://www.butterfliesofamerica.com/L/blepolenis_b_batea_types.htmH\u00fcbner, 1820http://eol.org/pages/149491/overviewhttp://www.butterfliesofamerica.com/L/caligo_a_arisbe_types.htmhttp://eol.org/pages/4090003/overviewhttp://www.butterfliesofamerica.com/L/callicore_s_sorana_types.htmhttp://eol.org/pages/163630/overviewhttp://www.butterfliesofamerica.com/L/catonephele_a_acontius_types.htmhttp://eol.org/pages/156101/overviewhttp://www.butterfliesofamerica.com/L/colobura_d_dirce_types.htmhttp://eol.org/pages/158533/overviewhttp://www.butterfliesofamerica.com/L/dryas_i_iulia_types.htmhttp://eol.org/pages/12083351/overviewhttp://www.butterfliesofamerica.com/L/euptychoides_castrensis_types.htmhttp://eol.org/pages/146531/overviewhttp://www.butterfliesofamerica.com/L/eryphanis_r_reevesii_types.htmhttp://eol.org/pages/160030/overviewhttp://www.butterfliesofamerica.com/L/eresia_lansdorfi_types.htmhttp://eol.org/pages/176703/overviewhttp://butterfliesofamerica.com/L/eurema_a_albula_types.htmhttp://eol.org/pages/178177/overviewhttp://www.butterfliesofamerica.com/L/eurema_e_elathea.htmhttp://eol.org/pages/184116/overviewhttp://www.butterfliesofamerica.com/L/eurema_p_phiale_types.htmhttp://eol.org/pages/19949/overviewhttp://eol.org/pages/961111/overviewhttp://www.butterfliesofamerica.com/L/godartiana_muscosa_types.htmhttp://eol.org/pages/166283/overviewhttp://www.butterfliesofamerica.com/L/hamadryas_a_amphinome_types.htmhttp://eol.org/pages/166346/overviewhttp://www.butterfliesofamerica.com/L/hamadryas_f_februa_types.htmhttp://eol.org/pages/166361/overviewhttp://www.butterfliesofamerica.com/L/hamadryas_f_feronia_types.htmhttp://eol.org/pages/155098/overviewhttp://www.butterfliesofamerica.com/L/heliconius_besckei_types.htmhttp://eol.org/pages/151378/overviewhttp://www.butterfliesofamerica.com/L/heliconius_e_erato_types1.htmhttp://eol.org/pages/157369/overviewhttp://www.butterfliesofamerica.com/L/heliconius_e_ethilla_types.htmhttp://eol.org/pages/185550/overviewhttp://www.butterfliesofamerica.com/L/heliopetes_omrina_types.htmhttp://eol.org/pages/162840/overviewhttp://www.butterfliesofamerica.com/junonia_e_evarete_types.htmhttp://eol.org/pages/157257/overviewhttp://www.butterfliesofamerica.com/junonia_g_genoveva_types.htmhttp://eol.org/pages/264320/overviewhttp://www.butterfliesofamerica.com/L/leptotes_c_cassius_types.htmhttp://eol.org/pages/33170/overviewhttp://eol.org/pages/165801/overviewhttp://www.butterfliesofamerica.com/L/marpesia_c_chiron_types.htmhttp://eol.org/pages/29501563/overviewhttp://www.butterfliesofamerica.com/L/memphis_m_moruus_types.htmhttp://eol.org/pages/29514890/overviewhttp://www.butterfliesofamerica.com/L/memphis_otrere_types.htmhttp://eol.org/pages/23311886/overviewhttp://eol.org/pages/19988/overviewhttp://eol.org/pages/138539/overviewhttp://www.butterfliesofamerica.com/L/morpho_h_helenor_types.htmhttp://eol.org/pages/148144/overviewhttp://www.butterfliesofamerica.com/L/narope_cyllarus_types.htmhttp://eol.org/pages/150133/overviewhttp://www.butterfliesofamerica.com/L/opsiphanes_c_cassiae_types.htmhttp://eol.org/pages/147972/overviewhttp://www.butterfliesofamerica.com/L/opsiphanes_q_quiteria_types.htmhttp://eol.org/pages/148836/overviewhttp://www.butterfliesofamerica.com/L/opoptera_syme_types.htmhttp://eol.org/pages/138517/overviewhttp://www.butterfliesofamerica.com/L/pareuptychia_o_ocirrhoe.htmhttp://www.butterfliesofamerica.com/L/paryphthimoides_undulata_types.htmhttp://eol.org/pages/168780/overviewhttp://www.butterfliesofamerica.com/L/prepona_l_laertes_types.htmhttp://eol.org/pages/261603/overviewhttp://www.butterfliesofamerica.com/L/pseudolycaena_marsyas_types.htmhttp://eol.org/pages/183872/overviewhttp://butterfliesofamerica.com/L/pyrgus_orcus_types.htmhttp://eol.org/pages/170707/overviewhttp://www.butterfliesofamerica.com/L/siderone_g_galanthis_types.htmhttp://eol.org/pages/4068082/overviewhttp://butterfliesofamerica.com/siproeta_s_stelenes_types.htmhttp://eol.org/pages/164148/overviewhttp://www.butterfliesofamerica.com/L/smyrna_b_blomfildia_types.htmhttp://eol.org/pages/20450/overviewhttp://eol.org/pages/147615/overviewhttp://www.butterfliesofamerica.com/L/taygetis_acuta_types.htmhttp://eol.org/pages/146471/overviewhttp://www.butterfliesofamerica.com/L/taygetis_l_laches_types.htmhttp://eol.org/pages/139915/overviewhttp://www.butterfliesofamerica.com/L/taygetis_m_mermeria_types.htmhttp://eol.org/pages/146462/overviewhttp://www.butterfliesofamerica.com/L/taygetis_sylvia_types.htmhttp://eol.org/pages/164154/overviewhttp://www.butterfliesofamerica.com/L/temenis_l_laothoe_types.htmhttp://eol.org/pages/153045/overviewhttp://www.butterfliesofamerica.com/L/telenassa_t_teletusa_types.htmhttp://eol.org/pages/20632/overviewhttp://eol.org/pages/40034120/overviewhttp://www.butterfliesofamerica.com/L/yphthimoides_straminea_types.htmhttp://eol.org/pages/36076631/overviewhttp://www.butterfliesofamerica.com/L/zaretis_isidora_types.htmhttp://www.butterfliesofamerica.com/L/zaretis_isidora_types.htmhttp://butterfliesofamerica.com/zaretis_i_itys_types.htmThe present study showed greater richness of species than the studies performed by In southeastern Brazil Nymphalidae was the family with greatest richness; this diversity can be explained by the fact that this family has great diversity in morphology and habits, as well as in environments with varying vegetation types belongs to the subfamily studies , in addi studies . From thronments .Euremaalbula and Euremaelathea, also registered in this site, have cosmopolitan habits and great adaptations for disturbed areas (Morphohelenor, which was well sampled \u2013 51 individuals (Table ed areas . It is ils Table . These dMorphohelenor, Siproetastelenes, Heliconiuserato, and Heliconiusethilla coincided with the study realized at the University Campus Darcy Ribeiro, in an urbanized area in the Federal District (Euptychoidescastrensis is found in abundance in tropical rain forest environments, being registered in the states of S\u00e3o Paulo, Rio Grande do Sul, and Minas Gerais District . These aNymphalidae. The species shared among these two studies are: Adelphapleasure, Archaeopreponademophon, Asciamonuste, Callicoresorana, Coloburadirce, Dryasiulia, Eresialansdorfi, Eryphanisreevesii, Euremaalbula, Euremaelathea, Hamadryasamphinome, Hamadryasfebrua, Hamadryasferonia, Heliconiuserato, Heliconiusethilla, Junoniaevarete, Leptotescassius, Pareuptychiaocirrhoe, Pyrgusorcus, Preponalaertes, Siproetastelenes, Smyrnablomfildia, Taygetislaches, Temenislaothoe, Zaretisitys, and Yphthimoidesstraminea.There are no records of inventories for the Espinha\u00e7o mountain range within the state of Minas Gerais: this is the first published inventory for the region. This study and the only study in the Serra do Espinha\u00e7o about butterflies, conducted in Chapada Diamantina in Bahia - Brazil by It is emphasized that in this study the majority of butterflies species captured are typical of Cerrado and Atlantic Forest .Lepidoptera showed great research and conservation potential for the Serra do Intendente State Park. The biodiversity information should be made available for decision makers, specially for regions such as the one studied, which is currently threatened by mining, tourism, and housing developments.Further investigation on biodiversity should be conducted and motivated in this region. The group of"} +{"text": "Nucleic Acids Res. 2014 Dec 16;42(22):13887\u201396. doi: 10.1093/nar/gku1236.TamulaitieneG.SilanskasA.GrazulisS.ZarembaM.SiksnysV.Nucleic Acids Res. 2014 Dec 16;42(22):14022\u201330. doi: 10.1093/nar/gku1237.Throughout these articles, we have used the prefix \u2018N\u2019 (as in N.CglI and N.NgoAVII) to describe NTPases associated with the cognate restriction systems. However, this prefix is also used for nicking restriction enzymes as described in . To avoi"} +{"text": "The first author\u2019s name is incorrect. The correct name is: G. W. Wieger Wamelink.The third author\u2019s name is incorrect. The correct name is: Joep Y. Frissel.The correct citation is: Wamelink GWW, Goedhart PW, Frissel JY (2014) Why Some Plant Species Are Rare. PLoS ONE 9(7): e102674. doi:10.1371/journal.pone.0102674"} +{"text": "Phytophthora plurivora. PLoS ONE 9(1): e85368. doi:10.1371/journal.pone.0085368The third author's name is spelled incorrectly. The correct name is: Niklaus J. Gr\u00fcnwald. The correct citation is: Schoebel CN, Stewart J, Gr\u00fcnwald NJ, Rigling D, Prospero S (2014) Population History and Pathways of Spread of the Plant Pathogen"} +{"text": "The first author\u2019s name is spelled incorrectly. The correct name is: Nina B. Illarionova. The correct citation is: Illarionova NB, Brismar H, Aperia A, Gunnarson E (2014) Role of Na,K-ATPase \u03b11 and \u03b12 Isoforms in the Support of Astrocyte Glutamate Uptake. PLoS ONE 9(6): e98469. doi:10.1371/journal.pone.0098469"} +{"text": "The third author\u2019s name is spelled incorrectly. The correct name is: K. Suzanne Sherf. The correct citation is: Halliday DWR, MacDonald SWS, Sherf KS, Tanaka JW (2014) A Reciprocal Model of Face Recognition and Autistic Traits: Evidence from an Individual Differences Perspective. PLoS ONE 9(5): e94013. doi:10.1371/journal.pone.0094013"} +{"text": "The second author\u2019s name is spelled incorrectly. The correct name is: Antonia D\u00edaz-Moreno. The correct citation is:Escribano BM, D\u00edaz-Moreno A, Tasset I, T\u00fanez I (2014) Impact of Light/Dark Cycle Patterns on Oxidative Stress in an Adriamycin-Induced Nephropathy Model in Rats. PLoS ONE 9(5): e97713. doi:10.1371/journal.pone.0097713"} +{"text": "Cell Rep. 9(6): 2180\u20132191. doi: 10.1016/j.celrep.2014.11.035.In the Discussion, Khanna et al. is described as an accompanying manuscript. The correct reference for this paper is: Khanna A, Johnson DL, Curran SP (2014) Physiological Roles for"} +{"text": "Nucleic Acids Res. 2014 Dec 1;42(21):13269\u201379. doi: 10.1093/nar/gku1067.In the legends of Figures"} +{"text": "Sensors [http://www.mdpi.com/1424-8220/15/10/26281.The authors wish to add an Acknowledgments section to their paper published in Sensors , doi:10."} +{"text": "Nucl. Acids Res. 43 (13): 6207\u20136221. doi: 10.1093/nar/gkv603The authors wish to correct the full name of Elenora Leucci to Eleonora Leucci."} +{"text": "The second author\u2019s name is spelled incorrectly. The correct author\u2019s name is: Natalie C. Hahn.Rai1. PLoS ONE 9(8): e105077. doi:10.1371/journal.pone.0105077The correct citation is: Alaimo JT, Hahn NC, Mullegama SV, Elsea SH (2014) Dietary Regimens Modify Early Onset of Obesity in Mice Haploinsufficient for"} +{"text": "These \u03bc-1,3-bridging thio\u00adcyante anions bridge the CdII cations, forming chains that propagate parallel to the b axis.In the crystal structure of the title compound, [Cd(NCS) DOI: 10.1107/S1600536814024647/pk2535Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814024647/pk2535fig1.tif. DOI: Crystal structure of the title compound. Displacement ellipsoids are drawn at the 50% probability level. Symmetry code: i: ; ii: .1033510CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Scientific Reports 5: Article number: 1035310.1038/srep10353; published online: 06032015; updated: 09022015.In this Article, Fig. 5 is a duplication of Fig. 7. The correct Fig. 5 appears below as"} +{"text": "The third author\u2019s name is incorrect. The correct name is: Henricus G. Ruh\u00e9.2/3 Receptor Availability in Treatment Resistant Depression. PLoS ONE 9(11): e113612. doi:10.1371/journal.pone.0113612.The correct citation is: de Kwaasteniet BP, Pinto C, Ruh\u00e9 HG, van Wingen GA, Booij J, Denys D (2014) Striatal Dopamine DThe correct author contributions are: Conceived and designed the experiments: BDK CP JB HGR. Performed the experiments: BDK CP. Analyzed the data: BDK CP. Contributed reagents/materials/analysis tools: BDK CP JB. Wrote the paper: BDK CP JB HGR GVW DD."} +{"text": "The first author\u2019s name is spelled incorrectly. The correct name is: Yoav Atsmon-Raz. The correct citation is: Atsmon-Raz Y, Tannenbaum ED (2014) Repression/Depression of Conjugative Plasmids and Their Influence on the Mutation-Selection Balance in Static Environments. PLoS ONE 9(5): e96839. doi:10.1371/journal.pone.0096839"} +{"text": "The first author\u2019s name is spelled incorrectly. The correct name is: Karolina Skowronski. The correct citation is: Skowronski K, Andrews J, Rodenhiser DI, Coomber BL (2014) Genome-Wide Analysis in Human Colorectal Cancer Cells Reveals Ischemia-Mediated Expression of Motility Genes via DNA Hypomethylation. PLoS ONE 9(7): e103243. doi:10.1371/journal.pone.0103243"} +{"text": "The first, second and last links in the Data Availability Statement are incorrect. The correct Data Availability Statement is:The authors confirm that all data underlying the findings are fully available without restriction. Datasets used in this study have been deposited at opencontext.org and can be found at the following DOIs:http://dx.doi.org/10.6078/M76H4FBShttp://doi.org/10.6078/M7VX0DF7http://dx.doi.org/10.6078/M74Q7RW8http://dx.doi.org/10.6078/M7X34VD1http://dx.doi.org/10.6078/M7SB43PPhttp://dx.doi.org/10.6078/M70Z715Bhttp://dx.doi.org/10.6078/M7KS6PHVhttp://dx.doi.org/10.6078/M7D798BQhttp://dx.doi.org/10.6078/M7CC0XMThttp://dx.doi.org/10.6078/M73X84KXhttp://dx.doi.org/10.6078/M7S46PVNhttp://dx.doi.org/10.6078/M76H4FBShttp://dx.doi.org/10.6078/M78G8HM0.The DOIs are incorrect in the Illipinar and \u00c7atalh\u00f6y\u00fck Area TP site rows of Table S1. Please view the correct Table S1 here.Table S1List of sites used in this paper including phasing, chronologies, sample sizes, authors, and links to online databases where available. Assemblages in bold were part of this data sharing project. Only biometric data from the Pendik and Yenikap\u0131 assemblages were included in this project. Assemblages in regular typeface represent previously published data. doi:10.1371/journal.pone.0099845.s002(DOCX)Click here for additional data file."} +{"text": "Email nbudhoo@iapb.orgUseful and highly relevant news and updates (including events) related to eye health. For a free subscription, write to Neebha Budhoo: http://tinyurl.com/rehabcourse Email: vtc@gju.edu.joProfessional diploma and MSc in Vision Rehabilitation. For more information, visit www.health.uct.ac.za or email chervon.vanderross@uct.ac.zaShort courses, postgraduate diploma, and MPH Community Eye Health. Lions Medical Training Centre, Nairobi, Kenya. Small incision cataract surgery (SICS). Write to: The Training Coordinator, Lions Medical Training Centre, Lions SightFirst Eye Hospital, PO Box 66576-00800, Nairobi, Kenya. Tel: +254 20 418 32 39www.kcco.net or contact Genes Mng'anga atgenes@kcco.net and/or genestz@yahoo.comVisit Contact Anita Shah, ICEH, London School of Hygiene and Tropical Medicine, London WC1E 7HT, UK. admin@cehjournal.orgwww.cehjournal.orgwww.facebook.com/CEHJournal/https://twitter.com/CEHJournal Share your questions and experiences with us at correspondence@cehjournal.org Articles of up to 800 words considered.web@cehjournal.org or visit www.cehjournal.org/subscribeGet an email alert each time a new issue is published! Write to"} +{"text": "A reference is omitted from the article. All sentences in the Discussion section which mention \u201cin an accompanying paper\u201d should cite Flores-Sandoval et al., 2015.Marchantia polymorpha. PLoS Genet 11(5): e1005207. doi:10.1371/journal.pgen.1005207.The reference is: Flores-Sandoval E, Eklund DM, Bowman JL (2015) A Simple Auxin Transcriptional Response System Regulates Multiple Morphogenetic Processes in the Liverwort"} +{"text": "There are errors in the Author Contributions section. The correct contributions are: Conceived and designed the experiments: MB LB FB JC. Performed the experiments: MB EO LB. Analyzed the data: MB YB BB JZ FB JC. Contributed reagents/materials/analysis tools: BB JZ PD. Wrote the paper: MB YB MH AB RH FB JC."} +{"text": "The author contributions of the manuscript are incorrect, the correct contributions are:Conceived and designed the experiments: SE KH JK JL.Performed the experiments: KH MF CT SE AH.Analyzed the data: BA CT KH AH AK JL MF.Contributed materials and analysis tools: SE AH KH JK CT JL.Wrote the paper: SE KH JD MF JK JL.Preprocessed/organized raw food sales data: CT MF BA AK.Designed/wrote likelihood algorithm: SE JL.Designed and performed clustering analysis: AH CT.Proposed this use of retail data with public health data: MF JK."} +{"text": "The third author's name is spelled incorrectly. The correct name is: Sabrina D. Thiel. The correct citation is: Goltz D, Pleger B, Thiel SD, Villringer A, M\u00fcller MM (2013) Sustained Spatial Attention to Vibrotactile Stimulation in the Flutter Range: Relevant Brain Regions and Their Interaction. PLoS ONE 8(12): e84196. doi:10.1371/journal.pone.0084196"} +{"text": "The first author's name is displayed incorrectly. The correct name is: Geir Pedersen. The correct citation is: Pedersen G, Selsbakk JM, Theresa W, Sigmund K (2014) Testing Different Versions of the Affective Neuroscience Personality Scales in a Clinical Sample. PLoS ONE 9(10): e109394. doi:10.1371/journal.pone.0109394"} +{"text": "Unfortunately, the original version of this article containehttp://www.saglik.gov.tr/TR/dosya/1-97020/h/saglik-istatistik-yilligi-2013.pdf)\u201d should have read \u201cThe Turkish neonatal mortality rate is 0.4\u00a0% according to the Ministry of Health of Turkey (http://www.saglik.gov.tr/TR/dosya/1-97020/h/saglik-istatistik-yilligi-2013.pdf)\u201d.The sentence \u201cThe Turkish neonatal mortality rate is 0.04\u00a0% according to World Health Organization Reports and the Ministry of Health of Turkey (The corrected sentence has been included in full in this erratum."} +{"text": "There are errors in the Author Contributions section. The correct Author Contributions are: Conceived and designed the experiments: FX LM VM RL GL. Performed the experiments: FX LM. Analyzed the data: FX LM. Contributed reagents/materials/analysis tools: MZ GH. Wrote the paper: FX. Helped in designing the experiments: GH AD. Reviewed/edited the manuscript: LM AD RL GL."} +{"text": "The second author's name is spelled incorrectly. The correct name is: Zubair Kabir. The correct citation is: Ajagbe OB, Kabir Z, O'Connor T (2014) Survival Analysis of Adult Tuberculosis Disease. PLoS ONE 9(11): e112838. doi:10.1371/journal.pone.0112838"} +{"text": "During 2008\u20132012, 29.9% of adults aged \u226518 years currently employed in construction and 28.2% of those currently employed in mining were current smokers. Adults currently employed in construction were more likely than adults currently employed in manufacturing (23.3%), transportation/warehousing/utilities (23.2%), trade (22.0%), agriculture/forestry/fishing (18.6%), services (16.9%), or health care/social assistance (16.0%) to be current smokers.Sources: National Health Interview Survey, 2008\u20132012. Available at http://www.cdc.gov/nchs/nhis.htm.Reported by: Debra L. Blackwell, PhD, debra.blackwell@cdc.hhs.gov, 301-458-4103."} +{"text": "This particular report describes cellulose conversion to isobutanol.http://www.biotechnologyforbiofuels.com/content/6/1/59This report describes a yeast-bacterial consortium that transforms cellulose to ethanol.http://www.researchgate.net/publication/221885494_Towards_synthetic_microbial_consortia_for_bioprocessingThis Research Gate page describes a review article on broad principles for engineering microbial consortia for practical purposes.http://www.springer.com/chemistry/biotechnology/book/978-1-4939-0553-9This e-book provides an engineering view of using mixed prokaryotic species for carrying out industrially relevant processes.http://yoric.mit.edu/design-microbial-consortia-industrial-biotechnologyThis report from a conference proceedings describes a chemical engineering approach to designing multi-species systems for robust industrial biotechnology.http://www.ifac-papersonline.net/Detailed/64335.htmlTrichoderma and Saccharomyces species for degrading cellulose and converting the monomers to ethanol.This report describes an association between http://journals.sfu.ca/rncsb/index.php/csbj/article/view/csbj.201210017/188This report contains some nice examples of naturally-occurring consortia and synthetic, or engineered, consortia that carry out useful biotransformations.http://journal.frontiersin.org/Journal/10.3389/fmicb.2012.00203/fullThis Frontiers report largely focuses on mixed microbial populations for handling metals and metal remediation.http://www.eastscotbiodtp.ac.uk/using-synthetic-ecology-optimise-methanogenic-consortia-anaerobic-digestionBiogas production is a logical choice for investigating anaerobic microbial consortia that produce methane since methanogens are often highly metabolically interdependent with other microorganisms in natural settings.http://rsif.royalsocietypublishing.org/content/11/96/20140065This is a comprehensive review of co-cultures for biotechnology, has many illustrative figures, and over one hundred references.http://www.academia.edu/7450923/Consortia_of_cyanobacteria_microalgae_and_bacteria_Biotechnological_potentialThe benefits of cyanobacteria and heterotroph consortia are obvious, with the ability to harvest energy from light and having oxygen transfer from phototroph to heterotroph.http://2014.igem.org/Team:Edinburgh/logic/This IGEM project page describes the design of a control system for working with mixed populations of bacteria."} +{"text": "AbstractAlnusviridis (Chaix) DC., based on Betulaviridis Chaix (1785), has traditionally been attributed to green alders although it is based on a later basionym. Alnusalnobetula (Ehrh.) K. Koch based on Betulaalnobetula Ehrh. (1783) is the correct name for green alders. In light of the increasing use and recognition of the name Alnusalnobetula (Ehrh.) K. Koch in the literature. I herein propose new nomenclatural combinations to account for the Japanese and Chinese subspecies respectively: Alnusalnobetulasubsp.maximowiczii J. Chery and Alnusalnobetulasubsp.mandschurica J. Chery. Recent phylogenetic analyses place these two taxa in the green alder species complex, suggesting that they should be treated as infraspecific taxa under the polymorphic Alnusalnobetula.The name Alnusalnobetula (Ehrh.) K. Koch is an anemophilous shrub with carpellate catkins that develop into woody strobili. It has a circumpolar distribution with subspecies in Europe DC. has long been attributed to green alders; however a closer look at the literature reveals the name Alnusalnobetula (Ehrh.) K. Koch has priority Raus, Alnusalnobetulasubsp.sinuata (Aiton) Raus Lambinon & Kergu\u00e9len (The name priority . Appropron) Raus , and Alnergu\u00e9len . SubspecBetulaviridis Chaix dates from 1785 describing a shrub in which \u201cthe homeland is unknown to me\u201d (translated from German). In Betulaalnobetula Ehrh. reappeared.The confusion lies in the appropriate basionym of this taxon. The name Betula species were transferred to Alnus, authors were evidently unaware of the original 1783 publication of the name Betulaalnobetula Ehrh., so Betulaviridis Chaix was thought to be the older name and was taken to be the basionym for green alders. Alnusalnobetula Ehrh. has consistently been associated with the 1788 reproduced work and thus listed as a later synonym of Alnusviridis (Chaix) DC.As Alnusviridis (Chaix) DC. as a synonym of Alnusalnobetula (Ehrh.) K. Koch. Other databases seem to be waiting for formal action to account for all subspecies names. For example, USDA, Germplasm Resources Information Network K. Koch, based on Betulaalnobetula Ehrh. (1783) has priority over Alnusviridis (Chaix) DC., based on Betulaviridis Chaix (1786); nevertheless, Alnusviridis is retained here until all infraspecific taxa are accounted for under Alnusalnobetula\u201d. Other major databases have incomplete citation list for synonyms such as Fl. North Amer. North of Mexico Editorial Committee [http://www.efloras.org/flora_page.aspx?flora_id=1 \u2013 accessed 22.07.2015]. Flora Europea [http://rbg-web2.rbge.org.uk/FE/fe.html \u2013 accessed 22.07.2015] omits citations for green alder names.Major databases such as plantlist.org [accessed 22.07.2015], list the name Network [http://PageBreakAlnusmandshurica, Alnusfirma, Alnuspendula and Alnussieboldiana embedded within a greater polytomy that includes all other green alders Cherycomb. n.urn:lsid:ipni.org:names:77149153-1Alnusmaximowiczii Callier ex C.K. Schneid., Illustr. Handb. Laubholzk. 1: 122. 1904: typified by the plate accompanying the protologue (Basionym).Alnuscrispasubsp.maximowiczii Hult\u00e9n, Acta Univ. Lund. Avd. 2. 40(1): 590. 1944.Alnastermaximowiczii Czerep., Bot. Mater. Gerb. Bot. Inst. Komarova Akad. Nauk. S.S.S.R. 17: 97. 1955.Alnastercrispussubsp.maximowiczii Murai, Bull. Gov. Forest Exp.Sta.154: 62. 1963.Duschekiamaximowiczii Pouzar, Preslia 36: 339. 1964.Alnastermaximowiczii Czerep., Fl. Arct. URSS Fasc. 5, 133 in obs. 1966.Alnusviridissubsp.maximoviczii D. L\u00f6ve, Taxon 17: 89. 1968.Alnusviridissubsp.maximowiczii H. Ohba, Fl. Japan 2a: 27. 2006.Temperate Asia: Russian Federation - Khabarovsk, Kurile Islands, Primorye, Sakhalin; Japan - Hokkaido, Honshu; KoreaTaxon classificationPlantaeFagalesBetulaceae Cherycomb. n.urn:lsid:ipni.org:names:77149155-1Alnusfruticosavar.mandschurica Callier ex C.K. Schneid., Illustr. Handb. Laubholzk. 1:121. 1904: Lectotype: Nadelholzzone des Tschangpei-schan, immer vereinzelt, 1600\u20131800 m (Fenze 262); designated by Hand.-Mazz., not seen) (Basionym).Alnusfruticosavar.mandschurica Callier ex Kom., Acta Hort. Petr. 22: 59. 1903.Alnusfruticosavar.mandschuricaf.normalis Callier, Fedde, Rep. Spec. Nov. 10: 227. 1911.Alnusfruticosavar.mandschuricaf.grandifolia Callier, Fedde, Rep. Spec. Nov. 10: 227.1911.PageBreakAlnusmandschurica Hand.-Mazz., Oesterr. Bot. Z. 81: 306\u2013307.1932.Alnuscrispa(Aiton)Purshsubsp.mandshurica Hara, J. Fac. Sci. Univ. Tokyo III, -6, (2): 32. 1952.Alnusmandschuricavar.pubescens Baranov, in T. N. Liou, Illustrated Flora of Ligneous plants of N. E. China 206, t. 75, fig. 112, t. 76, figs 1\u20134. 1955.Duschekiamandschurica Pouzar, Preslia 36(4): 339. 1964.Alnastercrispa(Aiton)ssp.mandshurica Murai, Bull. Gov. For. Expt. Sta. Jap. 171: 34. 1964.Russian Federation: Khabarovsk, Primorye; China: Heilongjiang, Jilin, Liaoning, Nei Monggol; Korea"} +{"text": "The second author's name is spelled incorrectly. The correct name is: Lauren A. Howell. The correct citation is: Korrapati MC, Howell LA, Shaner BE, Megyesi JK, Siskind LJ, et al. (2013) Suramin: A Potential Therapy for Diabetic Nephropathy. PLoS ONE 8(9): e73655. doi:10.1371/journal.pone.0073655The Author Contributions should read: Conceived and designed the experiments: MCK LJS RGS. Performed the experiments: MCK LAH BES. Analyzed the data: MCK LAH JKM LJS RGS. Wrote the paper: MCK LJS RGS."} +{"text": "AbstractThe island of \u00d6land, at the southeast of Sweden, has unique geological and environmental features. The Station Linn\u00e9 is a well-known \u00d6land research station which provides facilities for effective studies and attracts researchers from all over the world. Moreover, the station remains a center for ecotourism due to extraordinary biodiversity of the area. The present paper is aimed to support popular science activities carried out on the island and to shed light on diverse geometrid moth fauna of the Station Linn\u00e9.Lepidoptera: Geometridae) collected on the territory of the station is presented. Images of moths from above and underside are shown. Of the totally 192 species registered for Sweden, 41 species (more than 21%) were collected in close proximity to the main building of the Station Linn\u00e9. Malaise trap sampling of Lepidoptera is discussed.As an outcome of several research projects, including the Swedish Malaise Trap Project (SMTP) and the Swedish Taxonomy Initiative (STI) conducted at the Station Linn\u00e9, a list of larentiine moths ( The island of \u00d6land, at the southeast of Sweden, is famous for its dominant environmental feature, an Ordovician limestone pavement, which is called the Stora Alvaret (= the Great Alvar). Alvars are semi-natural grasslands which have been formed and developed due to long periods of human influence, including grazing . The StoThe Swedish Malaise Trap Project (SMTP), funded by the Swedish Species Information Centre (ArtDatabanken), is based at the Station Linn\u00e9. The project aims to provide species determinations for the specimens obtained from Malaise traps sampling at a wide range of landscapes and habitats. For many groups, including geometrid moths, the final data release is still awaited. The present paper is aimed to present a first list of the larentiine moths collected at the Station Linn\u00e9.56.6186 N, 16.4989 E). The UV light trap was placed between the tree and shrub rows along a walking path, with a meadow on one side and a swampy area on the other side , a mercury vapor light trap (MV) and net sweeping (NS) by O. Schmidt in 2014 and 2015 (July 20-31) in the M\u00f6rbyl\u00e5nga kommun, Skogsby, Station Linn\u00e9 was checked and the larentiine moths identified. This Malaise trap was placed on a lawn, about 100 m north of the Alvar edge and was running from April 2007 until November 2008. A note is given for the species recorded from Malaise trap samples only.Furthermore, material collected as part of the SMTP in 2007 and 2008 using a Malaise trap (MF) located close to the main building of the Station Linn\u00e9 and publications by The genitalia of all small-sized moths were studied to correctly identify the species. The material was identified using the http://eol.org/pages/4031647/overviewhttp://www.lepiforum.de/lepiwiki.pl?Phibalapteryx_virgataFigs 4http://eol.org/pages/283762/overviewhttp://www.lepiforum.de/lepiwiki.pl?Cidaria_FulvataFigs 6http://eol.org/pages/277279/overviewhttp://www.lepiforum.de/lepiwiki.pl?Colostygia_olivataFigs 8http://eol.org/pages/278481/overviewhttp://www.lepiforum.de/lepiwiki.pl?Colostygia_PectinatariaFigs 10http://eol.org/pages/270324/overviewhttp://www.lepiforum.de/lepiwiki.pl?Cosmorhoe_OcellataFigs 12http://eol.org/pages/281849/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eulithis_PrunataFigs 14http://eol.org/pages/284564/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eulithis_MellinataFigs 16http://eol.org/pages/286201/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eulithis_TestataFigs 18http://eol.org/pages/4017307/overviewhttp://www.lepiforum.de/lepiwiki.pl?Gandaritis_PyraliataFigs 20http://eol.org/search?q=Plemyria+rubiginata&search=Gohttp://www.lepiforum.de/lepiwiki.pl?Plemyria_RubiginataFigs 22http://eol.org/pages/298019/overviewhttp://www.lepiforum.de/lepiwiki.pl?Thera_cognataFigs 24http://eol.org/pages/285123/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_absinthiatahttp://mothphotographersgroup.msstate.edu/species.php?hodges=7586.1Figs 26http://eol.org/search?q=Eupithecia+denotata&search=Gohttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_denotatahttp://www.euroleps.ch/seiten/s_art.php?art=geo_denotataFigs 28http://eol.org/pages/284131/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_exiguataFigs 30http://eol.org/pages/281026/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_icterataFigs 32http://eol.org/pages/283937/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_LinariataFigs 34http://eol.org/pages/287885/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_nanataFigs 36http://eol.org/search?http://eol.org/search?q=Eupithecia+pusillata&search=Gohttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_PusillataFigs 38http://eol.org/pages/292707/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_satyrataFigs 40http://eol.org/pages/286559/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_SubfuscataFigs 42http://eol.org/pages/287281/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_subumbrataFigs 44http://eol.org/pages/292925/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_succenturiataFigs 46http://eol.org/pages/285093/overviewhttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_tenuiataFigs 48https://en.wikipedia.org/wiki/Eupithecia_valerianatahttp://www.lepiforum.de/lepiwiki.pl?Eupithecia_ValerianataFigs 50http://eol.org/pages/4031644/overviewhttp://www.lepiforum.de/lepiwiki.pl?Pasiphila_ChloerataFigs 52http://eol.org/pages/277386/overviewhttp://www.lepiforum.de/lepiwiki.pl?Pasiphila_RectangulataFigs 54http://eol.org/pages/286763/overviewhttp://www.lepiforum.de/lepiwiki.pl?Hydriomena_furcataFigs 56http://eol.org/pages/284799/overviewhttp://www.lepiforum.de/lepiwiki.pl?Pelurga_ComitataFigs 58http://eol.org/pages/4012784/overviewhttp://www.lepiforum.de/lepiwiki.pl?Mesotype_DidymataFigs 60http://eol.org/pages/295004/overviewhttp://www.lepiforum.de/lepiwiki.pl?Perizoma_AlchemillataFigs 62http://eol.org/pages/296282/overviewhttp://www.lepiforum.de/lepiwiki.pl?Philereme_transversataFigs 64http://eol.org/search?http://eol.org/search?q=Philereme+vetulata&search=Gohttp://www.lepiforum.de/lepiwiki.pl?Philereme_vetulataFigs 66http://eol.org/pages/295986/overviewhttp://www.lepiforum.de/lepiwiki.pl?Scotopteryx_ChenopodiataFigs 68http://eol.org/pages/297593/overviewhttp://www.lepiforum.de/lepiwiki.pl?Pterapherapteryx_sexalataFigs 70http://eol.org/search?http://eol.org/search?q=Camptogramma+bilineata&search=Gohttp://www.lepiforum.de/lepiwiki.pl?Camptogramma_BilineataFigs 72http://eol.org/pages/276073/overviewhttp://www.lepiforum.de/lepiwiki.pl?Catarhoe_cuculataFigs 74http://eol.org/pages/279041/overviewhttp://www.lepiforum.de/lepiwiki.pl?Catarhoe_RubidataFigs 76http://eol.org/search?q=Epirrhoe+alternata&search=Gohttp://www.lepiforum.de/lepiwiki.pl?Epirrhoe_AlternataFigs 78http://eol.org/pages/285918/overviewhttp://www.lepiforum.de/lepiwiki.pl?Epirrhoe_HastulataFigs 80http://eol.org/pages/285474/overviewhttp://www.lepiforum.de/lepiwiki.pl?Epirrhoe_TristataFigs 82http://eol.org/pages/288630/overviewhttp://www.lepiforum.de/lepiwiki.pl?Xanthorhoe_ferrugataFigs 84Larentiinae are recorded for Sweden (http://www2.nrm.se/en/svenska_fjarilar/svenska_fjarilar.html). Bert Gustafsson listed 156 species occurring on \u00d6land . Currently, 41 species are recorded for the territory of the Station Linn\u00e9, which comprises 26.3% of the \u00d6land species and more than 21% of the Swedish larentiine fauna. Interestingly, 37 species were sampled during 22 nights of light trapping in summer 2014 and 2015, when the weather was not quite favorable for collecting. For comparison, a recent rapid biotic survey at a 365 hectare Charitable Research Reserve in Ontario (Canada) revealed only nine larentiine species and in the female genitalia (the length of the ductus bursae and the shape of the signum). The specimens require more detailed study.A series of specimens presumably belonging to the species e.g.Lepidoptera by means of malaise traps is a challenging method. Designed for Diptera and Hymenoptera, a malaise trap indeed effectively samples Lepidoptera, as they get trapped within the malaise tent, flying upward towards either the sun (during the day) or the moon (at night) see . HoweverSupplementary material 1Lepidoptera: Geometridae) of the Station Linn\u00e9List of larentiine moth species (Data type: occurencesFile: oo_64905.xlsSchmidt, O."} +{"text": "This erratum corrects article: \u201cSecondary reconstruction of vaginal stenosis using a posterior labial perforator based Falandry flap\u201d, The Pan African Medical Journal. 2015;21:185. doi:10.11604/pamj.2015.21.185.6559. The original version of this article was corr"} +{"text": "The second author\u2019s name is incorrect in the citation. The correct name is: van Sorge NM.The correct citation is:10.1371/journal.ppat.1004644Van Avondt K, van Sorge NM, Meyaard L (2015) Bacterial Immune Evasion through Manipulation of Host Inhibitory Immune Signaling. PLoS Pathog 11(3): e1004644. doi:"} +{"text": "Scientific Reports5: Article number: 1043810.1038/srep10438; published online: 07032015; updated: 09022015.In this Article, the Accession codes were omitted from the Additional Information section:http://www.ncbi.nlm.nih.gov/geo/). The NGS data were submitted under accession number GSE62010 (URL: http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE62017), The microarray data were submitted under accession number GSE62011 (URL: http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE62011) and the NanoString data were submitted under accession number GSE62017 (URL: http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE62017). These three accession numbers are linked as super series.Data availability: Accession codes: All the data generated and described in this article have been submitted to the NCBI Gene Expression Omnibus (GEO;"} +{"text": "Background. Applying quantitative morphological approaches in systematics research is a promising way to discover cryptic biological diversity. Information obtained through twenty-first century science poses new challenges to taxonomy by offering the possibility of increased objectivity in independent and automated hypothesis formation. In recent years a number of promising new algorithmic approaches have been developed to recognize morphological diversity among insects based on multivariate morphometric analyses. These algorithms objectively delimit components in the data by automatically assigning objects into clusters.Method. In this paper, hypotheses on the diversity of the Malagasy Nesomyrmex angulatus group are formulated via a highly automated protocol involving a fusion of two algorithms, (1) Nest Centroid clustering (NC clustering) and (2) Partitioning Algorithm based on Recursive Thresholding (PART). Both algorithms assign samples into clusters, making the class assignment results of different algorithms readily inferable. The results were tested by confirmatory cross-validated Linear Discriminant Analysis (LOOCV-LDA).Results. Here we reveal the diversity of a unique and largely unexplored fragment of the Malagasy ant fauna using NC-PART-clustering on continuous morphological data, an approach that brings increased objectivity to taxonomy. We describe eight morphologically distinct species, including seven new species: Nesomyrmex angulatus , N. bidentatussp. n., N. clypeatussp. n., N. deviussp. n., N. exiguussp. n., N. fragilissp. n., N. gracilissp. n., and N. hirtellussp. n.. An identification key for their worker castes using morphometric data is provided.Conclusions. Combining the dimensionality reduction feature of NC clustering with the assignment of samples into clusters by PART advances the automatization of morphometry-based alpha taxonomy. Madagascar, one of Earth\u2019s biodiversity hotspots , has a uNesomyrmex, may contain ten times more species than was previously described. This dramatic increase in suspected species is due to a profusion of microendemic species. Our approach to this taxonomic challenges is to apply a quantitative morphological approach in combination with modern algorithms to delineate species by statistical means.On Madagascar, the high rate of diversity and possible cryptic species and Partstering) using costering) and PARTstering) .Delimitations of clusters recognized by these exploratory analyses were tested via confirmatory Linear Discriminant Analysis (LDA) and Multivariate Ratio Extractor, MRA followinN. angulatus species-group comprises eight well-outlined clusters in the Malagasy zoogeographical region, all representing species; of these, seven taxa are new to science. The eight species outlined, Nesomyrmex angulatus , N. bidentatussp. n., N. clypeatussp. n., N. deviussp. n., N. exiguussp. n., N. fragilissp. n., N. gracilissp. n., and N. hirtellussp. n. are described or redefined here based on worker caste. We provide a combined key that includes both a traditional, character-based key and a numeric identification tool that helps readers resolve the most problematic cases.Multivariate evaluation of morphological data has revealed that the The final species hypotheses are corroborated by qualitative morphological characters. Combining NC clustering and PART has proved to be an efficient method to automate species delimitation in insect taxonomy.Ant samples used in this study comply with the regulations for export and exchange of research samples outlined in the Convention on Biological Diversity and the Convention on International Trade in Endangered Species of Wild Fauna and Flora. For field work conducted in Madagascar, permits to research, collect, and export ants were obtained from the Ministry of Environment and Forest as part of an ongoing collaboration between the California Academy of Sciences and the Ministry of Environment and Forest, Madagascar National Parks and Parc Botanique et Zoologique de Tsimbazaza. Approval Numbers: No. 0142N/EA03/MG02, No. 340N-EV10/MG04, No. 69 du 07/04/06, No. 065N-EA05/MG11, No. 047N-EA05/MG11, No. 083N-A03/MG05, No. 206 MINENVEF/SG/DGEF/DPB/SCBLF, No. 0324N/EA12/MG03, No. 100 l/fEF/SG/DGEF/DADF/SCBF, No. 0379N/EA11/MG02, No. 200N/EA05/MG02. Authorization for export was provided by the Director of Natural Resources.In the present study, 23 continuous morphometric traits were recorded in 378 worker individuals belonging to 266 nest samples collected in the Malagasy region.The material is deposited in the following institutions, abbreviations after type material investigated sections for each taxon.The full list of material morphometrically examined in this revision is listed in http://www.antweb.org). Images are linked to their specimens via the unique specimen code affixed to each pin (CASENT0486461). Online specimen identifiers follow this format: http://www.antweb.org/specimen/CASENT0486461.All images and specimens used in this study are available online on AntWeb , or a Leica DFC 425 camera in combination with the Leica Application Suite software (version 3.8). Distribution maps were generated in R via the Measurements were taken with a Leica MZ 12.5 stereomicroscope equipped with an ocular micrometer at a magnification of 100\u00d7. Measurements and indices are presented as arithmetic means with minimum and maximum values in parentheses. Body size dimensions are expressed in \u00b5m. Due to the abundance of worker specimens relative to queen and male specimens, the present revision is based on the worker caste only. Revision based on the study of the workers is further facilitated by the fact that the name-bearing type specimens of the vast majority of existing ant taxa belong to the worker caste. All measurements were made by the first author. For the definition of morphometric characters, earlier protocols were conCL: Maximum cephalic length in median line. The head must be carefully tilted to the position providing the true maximum. Excavations of hind vertex and/or clypeus reduce CL .FRS: Frontal carina distance. Distance between the frontal carinae immediately caudal of the posterior intersection points between the frontal carinae and the torular lamellae. If these dorsal lamellae do not laterally surpass the frontal carinae, the deepest point of the scape corner pits may be taken as the reference line. These pits take up the inner corner of the scape base when the scape is directed caudally and produce a dark triangular shadow in the lateral frontal lobes immediately posterior to the dorsal lamellae of the scape joint capsule ( capsule .ML (Weber length): Mesosoma length from caudalmost point of propodeal lobe to transition point between anterior pronotal slope and anterior pronotal shield. Preferentially measured in lateral view; if the transition point is not well defined, use dorsal view and take the centre of the dark-shaded borderline between pronotal slope and pronotal shield as anterior reference point. In gynes: length from caudalmost point of propodeal lobe to the most distant point of steep anterior pronotal face were used only in two species, http://purl.org/NET/mx-database). Taxonomic history and descriptions of taxonomic treatments were rendered from this software. Hymenoptera-specific terminology of morphological statements used in descriptions and identification key, and diagnoses are mapped to classes in phenotype-relevant ontologies (Hymenoptera Anatomy Ontology (HAO) phenotypes were compiled in mx (gy (HAO) via a URgy (HAO) ; for morIn verbal descriptions of taxa based on external morphological traits, recent taxonomic papers were conhttp://zoobank.org/. The LSID for this publication is: lsid:zoobank.org:pub:63B1A3E5-9E62-46AD-B594-6B3E83364D90. The online version of this work is archived and available from the following digital repositories: PeerJ, PubMed Central and CLOCKSS.The electronic version of this article in Portable Document Format (PDF) will represent a published work according to the International Commission on Zoological Nomenclature (ICZN), and hence the new names contained in the electronic version are effectively published under that code from the electronic edition alone. This published work and the nomenclatural acts it contains have been registered in ZooBank, the online registration system for the ICZN. The ZooBank LSIDs (Life Science Identifiers) can be resolved and the associated information viewed through any standard web browser by appending the LSID to the prefix The present statistical framework follows the procedure applied in Nest-centroid clustering (NC-clustering), and linear discriminant analysis (LDA) do not require special data preparation , hence raw data were applied for each of the statistical analyses. Data, however, are standardized for the multivariate ratio analysis (MRA) to prevent variables with large values from dominating the analysis . Variablcluster using nest samples as groups . The seccluster and MASSer . Classification hypotheses were imposed for all samples congruently classified by partitioning methods, while wild-card settings were given to samples that were incongruently classified by the two methods or proved to be outliers. To extract the best ratios for the easiest species separation in the key and diagnoses we applied multivariate ratio analysis (MRA), a modern statistical method based on principal component analysis (PCA) and linear discriminant analysis (LDA) .Eight clusters were identified by both clustering algorithms \u2018hclust\u2019 and \u2018kmeans\u2019 using function \u2018part\u2019. The pattern recognized by these partitioning algorithms can be fitted on the hierarchical structure seen on the dendrogram generated by NC clustering .The grouping hypotheses generated by the combination of hypothesis-free exploratory analyses was validated by Linear Discriminant Analysis with leave-one-out cross-validation (LOOCV-LDA). The overall classification success is 100% . The pheThe geographic distribution of each morphospecies corresponds to the known major areas of endemism in Madagascar and can Nesomyrmex angulatus , N. bidentatussp. n., N. clypeatussp. n., N. deviussp. n., N. exiguussp. n., N. fragilissp. n., N. gracilissp. n., N. hirtellussp. n..The eight species described here are as follows in alphabetic order: bidentatus-complex consists of two species: Nesomyrmex bidentatussp. n. and N. fragilissp. n.; the devius-complex includes two new species: N. deviussp. n., N. exiguussp. n., N. gracilissp. n. and N. hirtellussp. n.; while two species, N. angulatus and N. clypeatussp. n., form a complex of their own in the Malagasy zoogeographical region. Separation of species as well as complexes are convincingly supported by Multivariate Ratio Analyses. Morphometric data for species calculated on individuals are given in These species are grouped into four species complexes based on morphological similarity. The angulatus angulatus ilgii = latinodis = angulatus concolor = bidentatus Cs\u0151sz & Fisher sp. n.clypeatus Cs\u0151sz & Fisher sp. n.devius Cs\u0151sz & Fisher sp. n.exiguus Cs\u0151sz & Fisher sp. n.fragilis Cs\u0151sz & Fisher sp. n.gracilis Cs\u0151sz & Fisher sp. n.hirtellus Cs\u0151sz & Fisher sp. n.Note: absolute size is given in \u00b5m, indexes are dimensionless values minimum and maximum values are given in brackets. Classification power between couplet based on a certain character is calculated and percent value is given in parentheses. 1.clypeatusMedian clypeal notch present : Cdep Propodel spines short: SPST/CS = 0.286 (94.9%). In lateral view dorsal contour line of propodeal spine or tubercle continues in a flat transition into metasomal dorsum \u20134F. Post-5 (devius complex)Propodeal spines longer and acute: SPST/CS = 0.378 , (94.9%). In lateral view dorsal contour line of propodeal spine continues in bent transition to metasomal dorsum . Postpet4.bidentatusPropodeal spine very short: SPST CS = 0.264 (84.2%), forming blunt tubercle . Postocu-fragilisPropodeal spine moderately long: SPST CS = 0.307 (84.2%) and acute . Postocu5.6North Madagascar only, north of \u221215\u00b0 latitude\u2026 -7The middle and southern part of Madagascar, south of \u221215\u00b0 latitude\u2026 6.exiguusIn profile, petiolar node rounded, leaning backward . Postocu-gracilisIn profile, petiolar node rectangular . Postocu7.deviusPostocular distance longer, apical spine distance shorter: PoOC/SPST = 1.261 (92.2%). Combination of best ratios (PoOC/SPST and MW/PPH) yields 100% of classification success see ... deviu-hirtellusPostocular distance shorter, apical spine distance longer: PoOC/SPST = 1.122 (92.2%). Combination of best ratios (PoOC/SPST and MW/PPH) yields 100% of classification success see ... hirteNesomyrmex angulatus .Type material investigated.Leptothorax angulatus Mayr, 1862:739\u2014\u201cSinai\u201d [Egypt], collect. G.Mayr. Lectotype, designated by Bolton 1982: 324 ;Leptothorax angulatus r. ilgii Forel, 1894:82\u2014\u201cr. L. ilgii Forel typus Harar (Ilg)\u201d [Ethiopia] coll. Forel. Syntype ;Leptothorax latinodis Mayr, 1895:130\u2014\u201clatinodis\u201d G. Mayr Type, \u201cDelagoa Bay Mozambiqe\u201d, collect. G. Mayr. Holotype. , [morphometrically not investigated due to fractured mesosoma];Leptothorax angulatus var. concolor Santschi, 1914:107\u2014\u201cL. Goniothorax angulatus Mayr v. concolor Sant Type\u201d, Cote d\u2019Afrique or. angl. Ile de Mombasa Allaud & Jeannel Oct. 1911 St.3. Syntypes ;Description of workers. Body color: yellow; brown. Body color pattern: concolorous; only clava darker. Absolute cephalic size (\u00b5m): 688 , (n = 33). Cephalic length vs. maximum width of head capsule (CL/CWb): 1.258 . Postocular distance vs. cephalic length (PoOc/CL): 0.371 . Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: feebly convex. Vertex contour line in frontal view shape: straight; feebly convex. Vertex sculpture: main sculpture rugoso-reticulate, ground sculpture areolate. Gena contour line in frontal view shape: convex. Genae contour from anterior view orientation: converging; strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.280 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.325 . Longitudinal carinae on median region of frons: present. Smooth median region on frons: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.818 . Facial area of the scape absolute setal angle: setae absent, pubescence only. Median clypeal notch: absent. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus: present. Lateral carinae of clypeus: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 58\u201362\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.273 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.325 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.334 . Propodeal spine shape: straight; curving upward. Anterolateral pronotal corner: present. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.690 . Metanotal depression: absent. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Petiole width vs. absolute cephalic size (PEW/CS): 0.410 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture rogoso-reticulate. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.488 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.Diagnosis. Workers of N. angulatus can be convincingly separated from those of N. clypeatus based on the lack of median clypeal notch in the former species by having sharp anterolateral pronotal angles ;Paratypes: Eighteen workers, a single gyne and a male with the same locality data under CASENT codes: CASENT0486459, BLF6646, ; CASENT0486460, BLF6646, ; CASENT0486462, BLF6646, ; CASENT0486797, BLF6618, ; CASENT0486798, BLF6618, ; CASENT0486799, BLF6618, ; CASENT0488445, BLF6584(24), ; CASENT0746773, BLF6646, ;Description of workers. Body color: yellow. Body color pattern: concolorous. Absolute cephalic size (\u00b5m): 510 (n = 60). Cephalic length vs. maximum width of head capsule (CL/CWb): 1.277 . Postocular distance vs. cephalic length (PoOc/CL): 0.416 . Postocular sides of cranium contour frontal view orientation: converging anteriorly; parallel. Postocular sides of cranium contour frontal view shape: straight; feebly convex; convex. Vertex contour line in frontal view shape: straight; feebly convex. Vertex sculpture: main sculpture rugoso-reticulate, ground sculpture areolate. Gena contour line in frontal view shape: convex. Genae contour from anterior view orientation: converging; strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: absent. Eye length vs. absolute cephalic size (EL/CS): 0.264 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.399 . Longitudinal carinae on median region of frons: absent. Smooth median region on frons: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.665 . Facial area of the scape absolute setal angle: 0\u201315\u00b0. Median clypeal notch: absent. Ground sculpture of submedian area of clypeus: present. Median carina of clypeus: present. Lateral carinae of clypeus: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: cannot be measured. Spine length vs. absolute cephalic size (SPST/CS): 0.264 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.346 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.335 . Propodeal spine shape: triangular, blunt. Anterolateral pronotal corner: absent. Metanotal depression count: absent; inconspicuous if present. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Metapleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.397 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.462 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.Etymology. The name (bidentatus) refers to the short propodeal denticle pair of this species.Diagnosis. Workers of N. bidentatus differ from those of N. clypeatus by having no median clypeal notch D4 = + 1.973 T. fragilissp. n. (n = 42) D4 = \u2212 2.234 Distribution.Nesomyrmex bidentatus is distributed in rainforests and littoral rainforests along the coastline around the entirety of Madagascar. This species occurs syntopically with its sister species N. fragilis in the western Antsisarana region ;Paratypes: four workers with the same locality data under CASENT codes: CASENT0427944, BLF3007, ; CASENT0422553, BLF2968, ; CASENT0427970, BLF2970, ; CASENT0427971, BLF2971, ;Description of workers. Body color: yellow; brown. Body color pattern: concolorous, only clava darker. Absolute cephalic size (\u00b5m): 898 (n = 12). Cephalic length vs. maximum width of head capsule (CL/CWb): 1.076 . Postocular distance vs. cephalic length (PoOc/CL): 0.434 . Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: feebly convex. Vertex contour line in frontal view shape: straight; feebly convex. Vertex sculpture: main sculpture rugoso-reticulate, ground sculpture areolate. Gena contour line in frontal view shape: convex. Genae contour from anterior view orientation: strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: absent. Eye length vs. absolute cephalic size (EL/CS): 0.210 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.310 . Longitudinal carinae on median region of frons: absent. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.758 . Facial area of the scape absolute setal angle: setae absent, pubescence only. Median clypeal notch: present. Median clypeal notch depth vs. absolute cephalic size (Cdep/CS): 0.021 . Ground sculpture of submedian area of clypeus: present. Median carina of clypeus: present. Lateral carinae of clypeus: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 50\u201360\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.361 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.349 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.463 . Propodeal spine shape: straight; slightly bent. Anterolateral pronotal corner: present. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.773 . Metanotal depression count: absent. Dorsal region of mesosoma sculpture: rugose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Petiole width vs. absolute cephalic size (PEW/CS): 0.460 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture rogoso-reticulate. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.493 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.Etymology. The name (clypeatus) refers to the presence of an antero-median clypeal depression in this species, the characteristic found to be unique in this revisionary work.Diagnosis. This species cannot be confused with other taxa in this revisionary work based on the dark antennal club and the conspicuous median notch on the anterior clypeal border.Distribution. This species is endemic to the Malagasy region. It is known to occur in tropical dry forests and littoral forests of the northern, dry area of Madagascar ;Paratypes: fifteen workers, and 6 gynes with the same label data with the holotype under CASENT codes: CASENT0448818, BLF5777, ; CASENT0448819, BLF5777, ; CASENT0448823, BLF5777, ; CASENT0448824, BLF5777, ; CASENT0448825, BLF5777, ; CASENT0448829, BLF5777, ; CASENT0448830, BLF5777, ; CASENT0448831, BLF5777, ; CASENT0448832, BLF5777, ; CASENT0448833, BLF5777, ; CASENT0746772, BLF5777, ;Description of workers. Body color: yellow; brown. Body color pattern: concolorous. Absolute cephalic size (\u00b5m): 593 (n = 27). Cephalic length vs. maximum width of head capsule (CL/CWb): 1.188 . Postocular distance vs. cephalic length (PoOc/CL): 0.394 . Postocular sides of cranium contour frontal view orientation: converging anteriorly; parallel. Postocular sides of cranium contour frontal view shape: straight; feebly convex; convex. Vertex contour line in frontal view shape: straight; feebly convex. Vertex sculpture: main sculpture rugose, ground sculpture areolate. Gena contour line in frontal view shape: convex. Genae contour from anterior view orientation: converging; strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: absent. Eye length vs. absolute cephalic size (EL/CS): 0.263 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.415 . Longitudinal carinae on median region of frons: absent. Smooth median region on frons: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.632 . Facial area of the scape absolute setal angle: 0\u201315\u00b0. Median clypeal notch: absent. Ground sculpture of submedian area of clypeus: present. Median carina of clypeus: present. Lateral carinae of clypeus: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 42\u201347\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.340 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.371 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.430 . Propodeal spine shape: straight. Anterolateral pronotal corner: absent. Metanotal depression count: present. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Petiole width vs. absolute cephalic size (PEW/CS): 0.447 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture rugoso-reticulate. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.499 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.Etymology. The name (d\u0113vius = devious) refers to the relatively long path required to arrive at the current taxonomic situation of this species, caused by its superficial similarities to other taxa.Diagnosis. Workers of N. devius differ from those of N. clypeatus by having no median clypeal notch offers 92.2% success in determination between this species and N. Hirtellus, but a combination of two ratios (PoOC/SPST and MW/PPH) yields a safer determination ;Paratypes: seventeen workers, a single gyne and a male with the same locality data under CASENT codes: CASENT0077580, BLF10161, ; CASENT0746774, BLF10161, ; CASENT0077624, BLF10190, ; CASENT0077625, BLF10190, ; CASENT0077626, BLF10190, ; CASENT0077586, BLF10206, ; CASENT0077587, BLF10206, ;Description of workers. Body color: yellow; brown. Body color pattern: concolorous. Absolute cephalic size (\u00b5m): 586 (n = 84). Cephalic length vs. maximum width of head capsule (CL/CWb): 1.213 . Postocular distance vs. cephalic length (PoOc/CL): 0.408 . Postocular sides of cranium contour frontal view orientation: parallel; converging anteriorly. Postocular sides of cranium contour frontal view shape: straight; feebly convex; convex. Vertex contour line in frontal view shape: straight; feebly convex. Vertex sculpture: main sculpture rugose, ground sculpture areolate. Gena contour line in frontal view shape: convex. Genae contour from anterior view orientation: converging; strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: absent. Eye length vs. absolute cephalic size (EL/CS): 0.249 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.413 . Longitudinal carinae on median region of frons: absent. Smooth median region on frons: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.656 . Facial area of the scape absolute setal angle: 0\u201315\u00b0. Median clypeal notch: absent. Ground sculpture of submedian area of clypeus: present. Median carina of clypeus: present. Lateral carinae of clypeus: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 27\u201332\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.382 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.390 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.436 . Propodeal spine shape: straight; slightly bent. Anterolateral pronotal corner: absent. Metanotal depression count: present. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Petiole width vs. absolute cephalic size (PEW/CS): 0.437 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.516 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.Etymology. This name exiguus refers to the fact that this species is relatively easily to distinguish.Diagnosis. Workers of N. exiguus differ from those of N. clypeatus by having no median clypeal notch provides a safe opportunity for separation offers 94.5% success discriminating between this species and N. gracilis, and a combination of two ratios (PoOC/SPTI and CWb/ML) yields a safe determination when these were added as wildcards to minimize the chance of possible misclassifications. These individuals are most probably representatives of populations brought to these localities by people.dagascar . There idagascar . These sNesomyrmex fragilis Cs\u0151sz & Fisher sp. n..Type material investigated.Holotype worker: MADGAGASCAR: Prov. Antsisarana, Nosy Be, R\u00e9serve Naturelle Int\u00e9grale de Lokobe, 6.3 km 112\u00b0 ESE Hellville, 13.41933\u00b0S, 48.33117\u00b0E, 30 m, 19-24.iii.2001, collection code: BLF3496; CASENT0421396, Fisher et al. ;Paratypes: five workers and a single gyne with the same locality data under CASENT codes: CASENT0421397, BLF3496, ; CASENT0421395, BLF3496, ; CASENT0421398, BLF3482, ; CASENT0421399, BLF3482, ;Description of workers. Body color: yellow. Body color pattern: concolorous. Absolute cephalic size (\u00b5m): 539 (n = 42). Cephalic length vs. maximum width of head capsule (CL/CWb): 1.229 . Postocular distance vs. cephalic length (PoOc/CL): 0.404 . Postocular sides of cranium contour frontal view orientation: converging anteriorly; parallel. Postocular sides of cranium contour frontal view shape: straight; feebly convex; convex. Vertex contour line in frontal view shape: straight; feebly convex. Vertex sculpture: main sculpture rugoso-reticulate, ground sculpture areolate. Gena contour line in frontal view shape: convex. Genae contour from anterior view orientation: converging; strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: absent. Eye length vs. absolute cephalic size (EL/CS): 0.260 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.409 . Longitudinal carinae on median region of frons: absent. Smooth median region on frons: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.659 . Facial area of the scape absolute setal angle: 15\u201330\u00b0. Median clypeal notch: absent. Ground sculpture of submedian area of clypeus: present. Median carina of clypeus: present. Lateral carinae of clypeus: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 20\u201327\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.310 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.369 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.377 . Propodeal spine shape: triangular, blunt. Anterolateral pronotal corner: absent. Metanotal depression: present, inconspicuous. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Metapleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.407 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.470 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.Etymology. This name fragilis (=fragile) refers to the small size of this species.Diagnosis. Workers of N. fragilis differ from those of N. clypeatus by having no median clypeal notch .a region . A singlNesomyrmex gracilis Cs\u0151sz & Fisher sp. n..Type material investigated.Holotype worker: MADAGASCAR: Prov. Antsiranana, For\u00eat Ambato, 26.6 km 33\u00b0 Ambanja, 13\u00b027.87\u2032S, 48\u00b033.10\u2032E, 150 m, 8-11.xii.2004, collection code: BLF11548; CASENT0107191, Fisher et al. ;Paratypes: three workers, three gynes, and a male with the same locality data under CASENT codes: CASENT0763758, BLF11548, ; CASENT0107710, BLF11539, ; CASENT0107026, BLF11624, ; CASENT0107027, BLF11624, ;Description of workers. Body color: yellow; brown. Body color pattern: concolorous. Absolute cephalic size (\u00b5m): 620 , (n = 44). Cephalic length vs. maximum width of head capsule (CL/CWb): 1.199 . Postocular distance vs. cephalic length (PoOc/CL): 0.387 . Postocular sides of cranium contour frontal view orientation: parallel; converging anteriorly. Postocular sides of cranium contour frontal view shape: straight; feebly convex; convex. Vertex contour line in frontal view shape: straight; feebly convex. Vertex sculpture: main sculpture rugose, ground sculpture areolate. Gena contour line in frontal view shape: convex. Genae contour from anterior view orientation: converging; strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: absent. Eye length vs. absolute cephalic size (EL/CS): 0.252 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.413 . Longitudinal carinae on median region of frons: absent. Smooth median region on frons: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.647 . Facial area of the scape absolute setal angle: 0\u201315\u00b0. Median clypeal notch: absent. Ground sculpture of submedian area of clypeus: present. Median carina of clypeus: present. Lateral carinae of clypeus: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 40\u201345\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.399 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.393 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.489 . Propodeal spine shape: straight; slightly bent. Anterolateral pronotal corner: absent. Metanotal depression: present. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Petiole width vs. absolute cephalic size (PEW/CS): 0.454 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.500 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.Etymology. This name gracilis refers to the small, tiny appearance of this species.Diagnosis. Workers of N. gracilis differ from those of N. clypeatus by having no median clypeal notch offers 94.5% success in distinguishing between this species and N. exiguus, and a combination of two ratios (PoOC/SPTI and CWb/ML) yields a safe determination ;Paratypes: Twenty six workers and three gynes with the same locality data under CASENT codes:CASENT0457483, BLF6155, ; CASENT0457482, BLF6155, ; CASENT0457476, BLF6155, ; CASENT0457481, BLF6155, ; CASENT0457480, BLF6155, ; CASENT0457478, BLF6155, ; CASENT0457479, BLF6155, ; CASENT0457477, BLF6155, ; CASENT0439545, BLF6118, ; CASENT0439552, BLF6118, ; CASENT0439546, BLF6118, ; CASENT0439555, BLF6118, ; CASENT0439551, BLF6118, ; CASENT0439547, BLF6118, ; CASENT0439553, BLF6118, ; CASENT0439548, BLF6118, ; CASENT0439550, BLF6118, ; CASENT0439549, BLF6118, ; CASENT0439554, BLF6118, ; CASENT0457598, BLF6119, ; CASENT0457596, BLF6119, ; CASENT0457599, BLF6119, ; CASENT0457597, BLF6119, ;Description of workers. Body color: yellow; brown. Body color pattern: concolorous. Absolute cephalic size (\u00b5m): 592 , (n = 75). Cephalic length vs. maximum width of head capsule (CL/CWb): 1.187 . Postocular distance vs. cephalic length (PoOc/CL): 0.387 . Postocular sides of cranium contour frontal view orientation: parallel; converging anteriorly. Postocular sides of cranium contour frontal view shape: straight; feebly convex; convex. Vertex contour line in frontal view shape: straight; feebly convex. Vertex sculpture: main sculpture rugose, ground sculpture areolate. Gena contour line in frontal view shape: convex. Genae contour from anterior view orientation: converging; strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: absent. Eye length vs. absolute cephalic size (EL/CS): 0.272 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.412 . Longitudinal carinae on median region of frons: absent. Smooth median region on frons: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.667 . Facial area of the scape absolute setal angle: ca. 15\u00b0. Median clypeal notch: absent. Ground sculpture of submedian area of clypeus: present. Median carina of clypeus: present. Lateral carinae of clypeus: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 37\u201342\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.375 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.389 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.460 . Propodeal spine shape: straight; slightly bent. Anterolateral pronotal corner: absent. Metanotal depression: present. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Petiole width vs. absolute cephalic size (PEW/CS): 0.460 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.525 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.Etymology. The name hirtellus: hirtus (=hairy) + -ellus (diminutive) refers to the workers having short hairs.Diagnosis. Workers of N. hirtellus differ from those of N. clypeatus by having no median clypeal notch offers 92.2% success in distinguishing between this species and N. devius and a combination of two ratios (PoOC/SPST and MW/PPH) yields a safe determination and Multivariate Ratio Extractor, MRA in decimal format, altitude (Elevation) in meters a.s.l., collector\u2019s name, date and number of specimens investigated bearing the given CASENT number are provided. HT, Holotype; PT, paratype(s).Click here for additional data file.10.7717/peerj.1796/supp-2Table S2Hymenoptera-specific terminology of morphological statements used in descriptions, identification key, and diagnoses are mapped to classes in phenotype-relevant ontologies.Click here for additional data file.10.7717/peerj.1796/supp-3Table S3Morphometric data of 21 continuous morphometric traits of 378 individuals, plus two others applied to a small fragment of samples, is given in \u00b5m. CASENT code (casent), final species hypothesis (species), geographic coordinates and the name format as samples appear on the dendrogram (dendro-name) are also provided in the table. HT, Holotype; PT, paratype(s).Click here for additional data file.10.7717/peerj.1796/supp-4Supplemental Information 1R script of NC clustering and method PART implementing cluster methods \u201chclust\u201d and \u201ckmeans\u201d. Mark dendrogram function mapping the results of partitioning algorithm PART on the dendrogram is also added.Click here for additional data file."} +{"text": "Moral philosophy and psychology have sought to define the nature of right and wrong, and good and evil. The industrial turn of the twentieth century fostered increasingly technological approaches that conjoined philosophy to psychology, and psychology to the natural sciences. Thus, moral philosophy and psychology became ever more vested to investigations of the anatomic structures and physiologic processes involved in cognition, emotion and behavior - ultimately falling under the rubric of the neurosciences. Since 2002, neuroscientific studies of moral thought, emotions and behaviors have become known as \u2013 and a part of \u2013 the relatively new discipline of neuroethics. Herein we present Part 2 of a bibliography of neuroethics from 2002\u20132013 addressing the \u201cneuroscience of ethics\u201d \u2013 studies of putative neural substrates and mechanisms involved in cognitive, emotional and behavioral processes of morality and ethics.RefWorks citation management program.A systematic survey of the neuroethics literature was undertaken. Bibliographic searches were performed by accessing 11 databases, 8 literature depositories, and 4 individual journal searches, and employed indexing language for National Library of Medicine (NLM) Medical Subject Heading databases. All bibliographic searches were conducted using the This bibliography lists 397 articles, 65 books, and 52 book chapters that present (1) empirical/experimental studies, overviews, and reviews of neural substrates and mechanisms involved in morality and ethics, and/or (2) reflections upon such studies and their implications. These works present resources offering iterative descriptions, definitions and criticisms of neural processes involved in moral cognition and behaviors, and also provide a historical view of this field, and insights to its developing canon. Throughout much of recorded history, humans have sought to define the nature of right and wrong, and good and evil. Since antiquity, such questions have been the focus of moral philosophy. However, empirical and experimental movements of the late nineteenth century drew scientific attention to philosophical questions, and the queries of moral philosophy became the focus of the then nascent discipline of psychology. The industrial turn of the twentieth century fostered increasingly technological approaches that conjoined psychology to the natural sciences. Philosophical speculation, and psychological observation and experimentation became ever more rooted in, and vested to investigations of the anatomic structures and physiologic processes involved in cognition, emotion and behavior. Thus, studies of moral philosophy and moral psychology became the province of brain research, ultimately falling under the rubric of the neurosciences, which became firmly established as a titular field in the middle-to-late 1970s \u2009CR Seances Soc Biol Fil 1998, 192(6):1041-1049.Changeux JP: Rev Neurol 1994, 150(8-9):555-563.Laplane D. Paris: Association Decartes and J. Libbey Eurotext 1996.Huber G: The Significance of Free Will. New York: Oxford University Press 1996.Kane R: The Man with a Shattered World: The History of a Brain Wound. New York: Basic Books 1972.Luria AR: The Society of Mind. New York: Simon and Schuster 1986.Minsky M: Equality and Partiality. Oxford and New York: Oxford University Press 1991.Nagel T: Computation and Cognition: Toward a Foundation for Cognitive Science. Cambridge, Mass.: MIT Press 1984.Pylyshyn ZW: The Brain and Emotion. Oxford: Oxford University Press 1999.Rolls ET: Sources of the Self: The Making of Modern Identity. Cambridge, Mass.: Harvard University Press 1989.Taylor C: Since 2002, neuroscientific studies of moral thought, emotions and behaviors have become known as \u2013 and a part of \u2013 the relatively new discipline of neuroethics . How canThe late William Safire concluded his introductory remarks to the 2002 Dana Foundation conference \u201cNeuroethics \u2013Mapping the Field\u201d by congratulating the attendees for tackling \u201c\u2026the challenge of carving out a new territory for an old philosophical discipline\u201d by examihttp://www.ncbi.nlm.nih.gov/mesh/) indexing terms were used for generating bibliographies from PubMed and National Library of Medicine (NLM) Catalog. MeSH includes ethics-related terms developed for BIOETHICSLINE, a specialty database devoted to bioethical issues produced for NLM by the Kennedy Institute of Ethics from 1975\u20132000. Other databases were searched using descriptors specific to those databases. The searches were limited to work published from 2002 to 2013.Methods for systematically searching relevant literature devoted to neuroethics are identical to those utilized in Part 1 of this bibliography . Search http://pubmed.gov):PubMed )http://www.ncbi.nlm.nih.gov/nlmcatalog):The NLM Catalog )Academic Search Premier:Search Strategy: TX morality AND SU neurosciences AND SU philosophyProquest Research Library:Search Strategy: su AND su (neurosciences)JSTOR:Search Strategy: ab: AND ab:(neuroscience)http://www.worldcat.org).:WorldCat Philosopher's Index:Search Strategy: su AND su(neuroscience)Embase:Search Strategy: neuroscience:de AND morality:dehttp://www.drze.de/belit/).:BELIT :Search Strategy: [topic] morality neuroscienceshttp://dp.la/):Digital Public Library of America (DPLA) :Directory of Open Access Journals (DOAJ) :Hathi Trust Digital Library :European Library :Internet Archive :Globethics.net :Neuroethics-Wikiography ;Journal of Mental Health Ethics from McMaster University (http://www.jemh.ca/);Journal of Practical Ethics (http://www.jpe.ox.ac.uk/) from the Oxford Uehiro Centre for Practical Ethics at the University of Oxford; andPhilosophers\u2019 Imprint from the University of Michigan (http://www.philosophersimprint.org/).As previously noted , open acRefWorks citation manager program was utilized to eliminate duplicate reference citations.As in Part 1 of this bibliography , the RefThick concepts and the moral brain.Euro J Soc 2011, 52(1):143\u2013172. doi:10.1017/s0003975611000051.Abend G: What the science of morality doesn\u2019t say about morality.Philos Soc Sci 2013, 43(2):157\u2013200. doi:10.1177/0048393112440597.Abend G: The social brain: neural basis of social knowledge.Annu Rev Psychol 2009, 60: 693:716. doi:10.1146/annurev.psych.60.110707.163514.Adolphs R: Cognitive neuroscience of human social behaviour.Nat Rev Neurosci 2003, 4(3): 165\u2013178. doi:10.1038/nrn1056.Adolphs R: Still afraid of needy post-persons.J Med Ethics 2013, 39(2):81\u201383. doi:10.1136/medethics-2012-101095.Agar N: Why is it possible to enhance moral status and why doing so is wrong?J Med Ethics 2013, 39(2):67\u201374. doi:10.1136/medethics-2012-100597.Agar N: Pr\u00e9cis of breakdown of will.Behav Brain Sci 2005, 28(5):635\u2013673. doi:10.1017/S0140525X05000117.Ainslie G: Ethical concepts and future challenges of neuroimaging: an Islamic perspective.Sci Eng Ethics 2012, 18(3):509\u2013518. doi:10.1007/s11948-012-9386-3.Al-Delaimy WK: Neuroscience, free will and moral responsibility.TRAMES-J Humanit Soc 2011, 15(2):147\u2013155. doi:10.3176/tr.2011.2.03.\u00c1rnason G: Neuroimaging, uncertainty, and the problem of dispositions.Camb Q Healthc Ethics 2010, 19(2):188\u2013195. doi:10.1017/S0963180109990454.\u00c1rnason G: Neurofunctional correlates of esthetic and moral judgments.Neurosci Lett 2013, 534:128\u2013132. doi:10.1016/j.neulet.2012.11.053.Avram M et al.: The biological foundations of culture and morality.Rendiconti Lincei 2008, 19 (2):189\u2013204.doi:10.1007/s12210-008-0011-y.Azzone GF: Neurosciences et neuro\u00e9thique: qui se ressemble s\u2019assemble.Rev Med Suisse 2005, 1(34):2225\u20132229.Baertschi B: Neurosciences et responsabilit\u00e9 morale: un argument en faveur du compatibilisme.Revue de Theologie et de Philosophie 2011, 143(3):257\u2013272.Baertschi B: Sociotopy in the temporoparietal cortex: common versus distinct processes.Soc Cogn Affect Neurosci 2010, 5(1):48\u201358. doi:10.1093/scan/nsp045.Bahnemann M et al.: Virtue essentialism, prototypes, and the moral conservative opposition to enhancement technologies: a neuroethical critique.AJOB Neurosci 2011, 2(2):31\u201338. doi:10.1080/21507740.2011.556918.Banja J: Modelling autonomy: simulating the essence of life and cognition. 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In From DNA to Social Cognition. Hoboken, N.J.: Wiley-Blackwell; 2012:111\u2013122.Koenigs M: Mind metaphors, neurosciences and ethics. In The New Brain Sciences: Perils and Prospects. Edited by Dai Rees, Steven Rose. New York: Cambridge University Press; 2004:71\u201387.Kollek R: The emerging theory of motivation. In The Oxford Handbook of Philosophy and Neuroscience. Edited by John Bickle. Oxford, U.K.: Oxford University Press; 2009:381\u2013418.Landreth A: The neuroscience of free will. In his Neuroethics: Challenges for the 21st Century. New York: Cambridge University Press; 2007:222\u2013257.Levy N: Commentary on Churchland. In Human Dignity and Bioethics: Essays Commissioned by the President's Council on Bioethics. Washington, DC: President\u2019s Council on Bioethics; 2008: 122\u2013125.Meilaender G: Free will and neuroscience. In his Free Will and Luck. Oxford, U.K.: Oxford University Press, 2006:30\u201348.Mele A: Do we ever really act? In The New Brain Sciences: Perils and Prospects. Edited by Dai Rees, Steven Rose. New York: Cambridge University Press; 2004:17\u201333.Midgley M: Neuroscience and morality: moral judgments, sentiments, and values. In Personality, Identity, and Character. Edited by Darcia Narvaez, Daniel K. Lapsley. New York: Cambridge University Press; 2009:106\u2013135.Moll J, de Oliveira-Souza R, Zahn R: How can evolution and neuroscience help us understand moral capacities? In The Moral Brain: Essays on the Evolutionary and Neuroscientific Aspects of Morality. Edited by Jan Verplaetse, Jelle De Schrijver, Sven Vanneste, Johan Braeckman. Netherlands: Springer; 2009 New York: Springer; 2009:201\u2013209.Nesse RM: Consciousness and the limits of neurobiology. In The New Brain Sciences: Perils and Prospects. Edited by Dai Rees, Steven Rose. New York: Cambridge University Press; 2004:59\u201370.Rose H: Having a brain, being a mind. In his The Future of the Brain: The Promise and Perils of Tomorrow's Neuroscience. Oxford, U.K.: Oxford University Press; 2006:137\u2013168.Rose S: What\u2019s \u201cneu\u201d in neuroethics. In The Oxford Handbook of Philosophy and Neuroscience. Edited by John Bickle. Oxford, U.K.: Oxford University Press; 2009:454\u2013472.Roskies A: A case study in neuroethics: the nature of moral judgment. In Neuroethics: Defining the Issues in Theory, Practice, and Policy. Edited by Judy Illes. Oxford, U.K.: Oxford University Press; 2006:17\u201332.Roskies A: \u00dcber einen m\u00f6glichen normativen beitrag der Moralphysiologie. In: Brauchen wir eine neue Moral?: Herausforderungen der Ethik durch die Neurowissenschaft. Edited by Gilbert Scharifi. Paderborn: Mentis; 2011:181\u2013198.Schleim S: The neurobiological basis of morality. In The Oxford Handbook of Neuroethics. Edited by Judy Illes, Barbara J. Sahakian. Oxford, U.K.: Oxford University Press; 2013:33\u201358.Suhler C, Churchland PS: Neuroscience and morality. In his Hardwired Behavior: What Neuroscience Reveals about Morality. New York: Cambridge University Press; 2005:1\u201311.Tancredi L: Morality and the mind. In his Hardwired Behavior: What Neuroscience Reveals about Morality. New York: Cambridge University Press; 2005:12\u201324.Tancredi L: The moral brain. In his Hardwired Behavior: What Neuroscience Reveals about Morality. New York: Cambridge University Press; 2005:34\u201345.Tancredi L: Creating a moral brain. In Hardwired Behavior: What Neuroscience Reveals about Morality. New York: Cambridge University Press; 2005:162\u2013176.Tancredi L: The bad brain: biology of moral thinking. In The Variables of Moral Capacity. Edited by David C. Thomasma and David N. Weisstub. Dordrecht: Kluwer; 2004:235\u2013257.Tancredi LR: Being in the world: neuroscience and the ethical agent. In Neuroethics: Defining the Issues in Theory, Practice, and Policy. Edited by Judy Illes. Oxford, U.K.: Oxford University Press; 2006:61\u201374.Zoloth L: bioethics@georgetown.edu for subsequent inclusion as commentary/addenda to this work.Despite our best efforts to amass as complete a bibliography of the past 10\u00a0years\u2019 neuroethics literature as possible, automated indexing and other technical issues can affect the retrieval of documents. However, this need not constrain the capability of this document to provide a valuable nexus in, and for the discipline. As consistent with Part 1 of this series, we conceive of this bibliography as a participatory endeavor, and request that the readership contribute to this effort by adding any missing citations to the online comments section of this bibliography. Citations also can be emailed directly to the bibliographic manager at: To be sure, with advances in neuroscientific capabilities and expanding use of neuroscientific techniques and technologies in medicine, arguments are being made to address ethical issues generated by brain research , promptiIn addition to implications for clinical care, neuroscientific studies of cognition, emotion and behavior can be \u2013 and are increasingly \u2013 leveraged in legal and social contexts, which must be considered on an international scale ,10. How,"} +{"text": "There are errors in the Author Contributions. The publisher apologizes for the errors. The correct contributions are:Conceived and designed the experiments: JLi X-zS. Performed the experiments: WZ JWL RX CZ QP XC LH YC. Analyzed the data: WZ JLi X-zS. Contributed reagents/materials/analysis tools: SL YC JY XL. Wrote the manuscript: JLi X-zS. Supervision of research: SL LH JY JLi X-zS."} +{"text": "The third author\u2019s name is spelled incorrectly. The correct name is: Khek Yu Ho. The correct citation is:10.1371/journal.pone.0148035Do TN, Seah TET, Ho KY, Phee SJ (2016) Development and Testing of a Magnetically Actuated Capsule Endoscopy for Obesity Treatment. PLoS ONE 11(1): e0148035. doi:"} +{"text": "References 32, 33, 36, and 41 are incomplete. Please view the corrected references here.http://209.61.208.233/LinkFiles/Dengue_CFR_Dengue_85-06.pdf.32. WHO SEARO. [cited 2014 January 3]; Available from: http://apps.who.int/gho/data/node.main.692.33. WHO Global Health Observatory Data Repository: Life expectancy. [cited 2014 January 14]; Available from: http://thailand.prd.go.th/view_news.php?id=6594&a=2.36. Office of the Prime Minister In: Thailand, editor. Thailand. [cited 2014 February 11]; Available from: http://www.uis.unesco.org/DataCentre/Pages/country-profile.aspx?code=THA®ioncode=4051541. UNESCO Institute for Statistics. [cited 2014 March 20]; Available from:"} +{"text": "Regarding the article \"A multiprofessional perspective on the principal barriers touniversal health coverage and universal access to health in extremely poor territories: thecontributions of nursing\", with DOI number: 10.1590/1518-8345.1042.2688, published in theRev. Latino-Am. Enfermagem. 2016;24:e2688, page 1:Where was written:\"Rev. Latino-Am. Enfermagem. 2016;24:e2688\"Now Read:\"Rev. Latino-Am. Enfermagem. 2016;24:e2795\"Regarding the article \"Access the Unified Health System actions and services from theperspective of judicialization\", with DOI number: 10.1590/1518-8345.1012.2689, published inthe Rev. Latino-Am. Enfermagem. 2016;24:e2689, page 1:Where was written:\"Rev. Latino-Am. Enfermagem. 2016;24:e2689\"Now Read:\"Rev. Latino-Am. Enfermagem. 2016;24:e2797\"Regarding the article \"Health-related quality of life as a predictor of mortality inpatients on peritoneal dialysis\", with DOI number: 10.1590/1518-8345.0786.2687, publishedin the Rev. Latino-Am. Enfermagem. 2016;24:e2687, page 1:Where was written:\"Rev. Latino-Am. Enfermagem. 2016;24:e2687\"Now Read:\"Rev. Latino-Am. Enfermagem. 2016;24:e2794\""} +{"text": "In the crystal there no inter\u00admolecular hydrogen bonds.In the title compound, C DOI: 10.1107/S2056989015010476/zs2333Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989015010476/zs2333Isup3.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989015010476/zs2333fig1.tif. DOI: Mol\u00adecular configuration and atom numbering scheme for the title compound with displacement ellipsoids drawn at the 50% probability level. Hydrogen atoms are omitted for clarity.Click here for additional data file.10.1107/S2056989015010476/zs2333fig2.tif. DOI: Crystal packing diagram of the title compound.Click here for additional data file.10.1107/S2056989015010476/zs2333fig3.tif. DOI: Synthetic scheme for the title compound (I).1062075CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Data is presented in support of model-based total evidence (MBTE) phylogenetic reconstructions of the Neotropical clade of Gymnotiformes \u201cModel-based total evidence phylogeny of Neotropical electric knifefishes \u201d Specifications TableValue of the data\u2022Data summary for the most comprehensive phylogenetic study of Gymnotiformes to date, including an ingroup taxon sampling of 33 (94%) recognized genera and 120 (57%) of all valid species.\u2022New molecular sequences for 149 specimens and descriptions of morphological characters for 166 specimens.\u2022Supermatrix comprised of six genes (5277\u00a0bp) and 223 morphological characters used to reappraise relationships of GymnotiformesDataThe data provided below include supporting information to replicate phylogenetic analyses of Tagliacollo et al. Experimental design, materials and methodsCarassius auratus, Erythrinus erythrinus, Serrasalmus rhombeus, Cyphocharax festivus, Charax tectifer, Pseudostegophilus nemurus, Brachyplatystoma juruense, Dianema longibarbis, Pterygoplichthys multiradiatus. Ingroup taxon samples were chosen by using a clade-based approach to maximize the representation of phylogenetic diversity in Gymnotiformes.Outgroups were chosen to cover a broad spectrum of ostariophysan diversity in terms of clade representation. Outgroups included nine major lineages of Otophysi represented by: Ingroup species are comprised of representatives of all major gymnotiform clades, including 33 of 35 (94%) recognized genera and 120 of 218 (55%) of all currently valid species. Voucher specimens for tissue samples were identified either directly by the authors, directly by curators and collection managers at contributing institutions, or by exchange of photographs. Species identifications of Genbank sequences were not reevaluated. Molecular vouchers and GenBank accession numbers are presented in \u00ae 96 Tissue kit (Macherey-Nagel). Fragments of the mitochondrial genes 16S rRNA (16S-mit), Cytochrome Oxidase subunit I (COI-mit), Cytochrome B (CytB-mit), and the nuclear gene Zic family member 1 (ZIC-nuc) were amplified by one round of polymerase chain reaction (PCR), which was carried out in a volume of 25.0\u00a0\u03bcl consisting of: 2.5\u00a0\u03bcl of 10x Taq Buffer, 2.0\u00a0\u03bcl of dNTP mixture at 10\u00a0mM each, 1.5\u00a0\u03bcl of 50\u00a0mM MgCl2, 1.0\u00a0\u03bcl of each primer at 5\u00a0\u03bcM, 0.2\u00a0\u03bcl of Platinum\u00ae Taq DNA Polymerase, 2.0\u00a0\u03bcl of template DNA (~50\u00a0ng), and 15.8\u00a0\u03bcl of double-distilled H2O. Fragments of the nuclear gene Recombination-Activating gene 2 (RAG2-nuc) and Recombination-Activating gene 1 (RAG1-nuc) were amplified by nested-PCRs. Each round of the two PCR was carried out in a volume of 25.0\u00a0\u03bcl consisting of: 2.5\u00a0\u03bcl of 10x Taq Buffer, 2.0\u00a0\u03bcl of dNTP mixture at 10\u00a0mM each, 2.0\u00a0\u03bcl of 50\u00a0mM MgCl2, 1.5\u00a0\u03bcl of each primer at 5\u00a0\u03bcM, 0.2\u00a0\u03bcl of Platinum\u00ae Taq DNA Polymerase, 2.0\u00a0\u03bcl of template DNA (~50\u00a0ng), and 14.8\u00a0\u03bcl of double-distilled H2O. Cycles of PCR for the mitochondrial genes consisted of five steps: (1) 60s for enzyme activation at 94\u00a0\u00b0C, (2) 30s of denaturation at 94\u00a0\u00b0C, (3) 60s of annealing at 56\u00a0\u00b0C (16S-mit), 54\u201358\u00a0\u00b0C (COI-mit), or 50\u201352\u00a0\u00b0C (CytB-mit), (4) 80s of extension at 72\u00a0\u00b0C, and (5) 300\u00a0s of extension at 72\u00a0\u00b0C. The steps 2\u20134 were repeated 35 times. Cycles of PCR for the nuclear genes consisted of six steps: (1) 60s for enzyme activation at 94\u00a0\u00b0C, (2) 30s of denaturation at 94\u00a0\u00b0C, (3) two start cycles of 60s each at 56\u00a0\u00b0C, 50\u00a0\u00b0C, 52\u00a0\u00b0C, 54\u00a0\u00b0C and 54\u00a0\u00b0C, 50\u00a0\u00b0C 52\u00a0\u00b0C, 56\u00a0\u00b0C (ZIC-nuc), (4) 60s of annealing at 50\u00a0\u00b0C and 52\u00a0\u00b0C (ZIC-nuc) and (5) 80s of extension at 72\u00a0\u00b0C, and (6) 300s of extension at 72\u00a0\u00b0C. The steps 2, 4 and 5 were repeated 35 times. PCR products were visually identified on a 1% agarose gel. Sequencing was held at Beckman Coulter Genomics Facility. The list of primers is shown in Genomic DNA was extracted from tissues, fins or livers of specimens preserved in pure ethanol with the NucleoSpinForward and reverse sequences were assembled in Geneious 5.5.6. The IUPAC ambiguity code of nucleotides was applied in cases where nucleotide identity was dubious. We combined newly generated data with available sequences from previous studies 1.Body shape 1.0: body laterally compressed, body width at pectoral fin base less than 70% its depth. 1: Body cylindrical or subcylindrical, roughly circular in cross section, body depth at pectoral girdle approximately equal to its width.2.Body shape 2.0: body laterally compressed. 1: Body dorsoventrally flattened. Newly coded herein.3.Body shape profile. \u201cBody Depth,\u201d character 2 in Albert, 2001. 0: Body relatively deep in profile, depth at pectoral girdle more than 11% total length. 1: Body elongate, slender, depth less than 11% total length.4.Snout length short. 0: preorbital length about one-third total head length in mature specimens. 1: Snout short, preorbital length less than one-third total head length -Fig. 13.5.Snout long. 0: Length of the snout about one-third total head length in mature specimens. 1: Snout elongate, frontal, vomer and anterior portion of parasphenoid elongate; preorbital length longer than one-third total head length or greater in mature specimens pale bands with straight margins of alternating high and low melanophore density along lateral surface of body, oriented at an oblique angle to longitudinal body axis -Fig. 1. 17.Vertical pigment lines. 0: Vertical pigment lines absent along longitudinal body axis. 1: Thin vertical pigment lines present along longitudinal body axis. Newly coded herein.18.Vertical pigment bars. \u201cSaddle-shaped bars\u201d, character 5 in 19.Caudal Peduncle Spot. 0: Pale spot absent from base of caudal region. 1: Pale spot present at base of caudal region. Newly coded herein.20.Longitudinal lines. 0: Absent. 1: 2\u20133 thin dark lines extending posteriorly along the lateral body surface . 2: A wh21.Pigment contrast. 0: Body surface yellow or pale brown, lacking high contrast dark brown or black and white pigments. 1: High contrast dark brown or black and white pigments on body surface.22.Apteronotus albifrons species group. Newly coded herein.White posterior bars. 0: White or pale bars absent from caudal region. 1: White or pale bars present on caudal region as observed in members of the 23.White mid-sagittal pigments. 0: All mid-sagittal surfaces brown. 1: Mid-sagittal region of dorsal and mental surfaces bright white.24.Antorbital stripe. 0: Melanophores on snout distributed evenly. 1: Melanophores absent from narrow band passing lateral to nares -Fig. 90.25.Pigment distribution. 0: Pigments distributed homogeneously over body surface. 1: Black and white pigments distributed unevenly over body surface, darker and paler areas grading into one another; integument with a marbled or mottled appearance.26.Body translucence. 0: Body opaque in living and formalin-fixed specimens, lateral body surface covered with brown melanophores. 1: Body translucent in living specimens, yellow or pink hue in living specimens, yellow or hyaline in formalin-fixed specimens, melanophores sparse or absent on lateral body surface.27.Branchial opening. 0: Branchial opening extends along entire posterior margin of opercle, from isthmus to pectoral fin insertion. 1: Vertical extent of branchial opening restricted to region around pectoral fin base; ventral portion reduced by a dorsolateral continuation of epidermis from isthmus .28.Pseudotympanum. 0: Sixth vertebra partially covered by superior oblique. 1: Sixth vertebra not covered by superior oblique.29.Body squamation. 0: Scales present on body and head. 1: Body devoid of scales . 1: Tips of teeth directed anteriorly (recurved). 39.Premaxilla size. 0: Large. Lateral margin of premaxilla longer than lateral margin of maxilla, premaxilla extends posterodorsal to articulation of maxilla with autopalatine; articular surface of maxilla with autopalatine oriented anterodorsally. 1: Small. The anterodorsal orientation of the articular surface of the maxilla with the autopalatine is a consequence of the large size of the premaxilla and the associated posterior position of the maxilla.40.Premaxillary teeth. 0: Teeth present on premaxilla of adults. 1: Premaxillary dentition reduced or lost.41.Maxillary dentition. 0: A single row of 6\u201310 conical teeth in along outer margin of maxilla. 1: No teeth on maxilla.42.Maxilla size. \u201cOrientation and Shape of Maxilla\u201d-character 30 in 43.Anterior maxillary process. 0: Anterior process of maxilla absent. 1: Anterior process of maxilla extends anterior to articulation of maxilla and autopalatine, forming a tapered process, its ventral margin continuous with descending blade of maxilla; maxilla forked in lateral view. 2: Anterior process of maxilla cartilaginous; ventral margin of descending blade extends to articulation of maxilla with autopalatine, forming anterior border of maxilla; maxilla crescent shaped in lateral view , ventral margin concave.Dentary gracile. 0: Dentary robust, posterodorsal process rounded, ventral margin straight or slightly convex in lateral view. 1: Dentary gracile, posterodorsal process tapering to a point , a ventral extension of the medial surface of dentary where it covers the anterior portion of Meckel\u05f3s cartilage -Fig. 66.55.Dentary teeth size. 0: Teeth on posterior half of dentary roughly equal in size to anterior teeth. 1: Teeth on posterior half of dentary twice the size of anterior teeth. Newly coded herein, see de 56.M. Adductor mandibulae. 0: Belly of Adductor mandibulae muscle composed of muscle fibers and tendons. 1: Belly of Adductor mandibulae muscle with ossified intermuscular bones, oriented parallel to main axis of muscle fibers inserts exclusively on maxilla; two discrete muscle bundles insert on oral jaws; A1 inserts exclusively on maxilla, and A2 on dentary. 1: Additional insertion of A1 on first infraorbital broad and concave, with a medial groove located between two large anterolateral processes (forming articulation with premaxillae). 1: Anterior tip of mesethmoid small, anterodorsal surface narrow, with a median knob-shaped process directed anteriorly between two small lateral processes .68.Median septum of ventral ethmoid. 0: Portion of ventral ethmoid ossified within medial nasal septum approximately as long as deep; posterior margins of median septum and lateral process of ventral ethmoid approximately equal. 1: Ossified median septum of ventral ethmoid elongate in mature specimens, longer than deep, extending posterior to posterior margin of lateral process.69.Ventral ethmoid-Vomer. 0: Ventral ethmoid fused with vomer during growth. 1: Ventral ethmoid and vomer not fused in adults .70.Dermal vomer. 0: Dermal vomer extends from posterior margin of ventral ethmoid to parasphenoid. 1: Dermal vomer not ossified.71.Ethmoid cartilage. 0: Ethmoid cartilage anterior to lateral ethmoid longer than deep; antorbital region of snout longer than deep. 1: Ethmoid cartilage deeper than long; antorbital region of snout about as deep as long .72.Lateral ethmoid size. 0: Lateral ethmoid a large endochondral ossification in the antorbital region, arching laterally over Profundus (V1) nerve, with four margins; anterolateral process contacting ventral ethmoid, posteromedial process contacting parasphenoid, dorsomedial margin contacting frontal, and anteromedial margin contacting mesethmoid in 83.Sphenotic process. 0: Dorsolateral margin of sphenotic straight, anterior margin underlies frontal. 1: Dorsolateral margin of sphenotic bearing a transversely oriented crest or process exposed on dorsolateral edge, anterior margin not underlying frontal . 1: Accessory optic tract reduced or absent; discrete accessory optic nucleus not visible in sections .107.Integumental taste buds. 0: Taste buds present on head in characiforms, and over entire integumental surface in siluriforms; diameters of nerves V and VII equal to or larger than that of other cranial nerves in isthmal region; primary facial and vagal sensory nuclei larger than medial octaval nucleus. 1: Taste buds entirely absent from extra-oral integument; nerves V and VII smaller than other cranial nerves of isthmal region; primary facial and vagal sensory nuclei smaller than medial octaval nucleus.108.Schreckstoff/club cells. 0: Schreckstoff , club cells, and fright response present in Ostariophysi. 1: Schreckstoff, club cells, and fright response absent and central nervous systems 110.Ampullary organ rosettes. 0: Ampullary organs distributed individually in integument. 1: Ampullary organs clustered in rosettes 111.Active electroreception. 0: Passive, low frequency electroreception, used in predation; neural apparatus for detecting low frequency electric currents. 1: Electrogeneration and high frequency electroreception, used in communication and navigation (in addition to predation); neural apparatus for producing and detecting high frequency electric currents 112.Tuberous electroreceptors. 0: One class of tuberous electroreceptor organs. 1: Two classes of morphologically distinct tuberous electroreceptor organs 113.Preotic lateralis ganglia. 0: All preotic lateral line nerve ganglia form from separate placodes, their axonal bundles entering brain separately. 1: Anterodorsal, anteroventral, and preopercular\u2013mandibular lateral line nerve ganglia fused during ontogeny, their axons entering brain in a single bundle 114.Posterior lateral line nerve. 0: Posterior lateral line nerve with no accessory rami. 1: Posterior lateral line nerve with dorsal ramus 115.Lateral line afferents. 0: Lateral line afferents from electrosensory periphery intermingled as they course into the electrosensory lateral line lobe (ELL); fibers from different lateral line nerves not segregated. 1: Lateral line afferents fasciculated into discrete bundles; fibers from each lateral line nerve segregated from those of other lateral line nerves 116.Anterior extent of eminentia granularis. 0: Eminentia granularis (EG) of dorsal medulla well developed, extending to posterior pole of optic tectum. 1: EG small, its anterior margin not extending to contact optic tectum 117.Posterior EG. 0: Posterior margin of EG not extending to posterior margin of ELL. 1: Posterior lobe of EG well developed, wrapped around caudal lobe of cerebellum, its posterior margin extending to a vertical with posterior margin of ELL 118.Anterior corpus cerebellum. 0: Anterior lobe of corpus cerebellum large, extending anterior to midlength of optic tectum; cerebellum overlying commissure of optic tectum. 1: Anterior lobe of corpus cerebellum extending to midlength of optic tectum; commissure of optic tectum exposed on dorsal surface.119.Pacemaker nucleus. 0: Pacemaker nucleus of medulla oblongata small, positioned on midline of neuraxis, adjacent to medial longitudinal fasciculus; its ventral margin not contacting ventral aspect of medulla. 1: Pacemaker nucleus large, visible as a median, ovoid eminence on ventral surface of medulla; its ventral margin extending to medullary surface 120.Palatines. 0: Autopalatine totally or partially ossified, straight. 1: Autopalatine unossified, arched.121.Ectopterygoid. 0: Ectopterygoid ossified as a dentigerous element in membrane overlying ventral portion of endopterygoid. 1: Ectopterygoid and associated teeth absent . 1: Ascending process on lateral surface of endopterygoid; pterygocranial ligament ossified; base of ascending process situated approximately dorsal to articulation of quadrate with anguloarticular 123.Endopterygoid ascending process. 0: Ascending process of endopterygoid developed in juvenile stages of growth and retained into adult. 1: Small ascending process of endopterygoid in juveniles obliterated by growth along dorsal margin of bone; no endopterygoid process in adults.124.Endopterygoid anterior process. 0: Dorsal portion of pterygocranial ligament not ossified; base of ascending process of endopterygoid broader than its tip. 1: Entire extent of ligament ossified, forming a bony strut anterior to orbit; process equally as wide along most of its length125.Mesopterygoid dentition. 0: Numerous small teeth distributed in an irregular field on anterior portion of ventral surface of endopterygoid. 1: Few or no teeth on endopterygoid not attached to gill arches ossified, articulating with parasphenoid -Fig. 13.144.Pharyngobranchials. 0: Pharyngobranchials of third and fourth arches cartilaginous. 1: Pharyngobranchials of third and fourth arches ossified.145.Pharyngobranchial plates. 0: Four dentigerous plates present on posterior gill arches. 1: One dentigerous plate present on posterior gill arch of Albert, 2001. 0: Posterior surface of second ceratobranchial smooth. 1: Posterior surface of second ceratobranchial with a medially oriented process in Albert, 2001. 0: Lateral surface of fourth ceratobranchial smooth. 1: Lateral surface of fourth ceratobranchial with an anterolaterally oriented process ossified. 1: All five elements of basibranchial series unossified.159.Basibranchial one. 0: First (anterior) basibranchial elongate, width at midlength about same as at anterior and posterior ends. 1: First basibranchial foreshortened and broad, hourglass shaped, breadth at midlength narrower than at either end160.Urohyal head. 0: Anterior head of urohyal narrow, lateral surfaces flat. 1: Anterior head of urohyal large, with lateral ridges 161.Urohyal blade. 0: Posterior blade of urohyal ossified, extending posterior to fourth basibranchial. 1: Posterior blade of urohyal unossified, anterior head of urohyal positioned ventral to second basibranchial.162.Urohyal blade hyperossified. 0: Urohyal blade short, ossified to level of third basibranchial. 1: Urohyal blade long, ossified to level of fourth basibranchial.163.Posttemporal. 0: Posttemporal independent from supracleithrum in mature specimens. 1: Posttemporal fused with supracleithrum in mature specimens . 0: All hemal spines medial, fused with hemal arches in adult specimens; one to one correspondence between caudal vertebrae and associated hemal spines. 1: Three additional hemal spines positioned in hypaxial musculature posterior to body cavity, often lateral to unmodified hemal spines, rarely fused with hemal arches or parapophyses; irregular association with posterior thoracic and anterior caudal vertebrae.178.DHS anterior series. 0: Three DHSs in hypaxial musculature immediately posterior to body cavity. 1: Anterior series of 8\u201314 DHSs in hypaxial musculature lateral to body cavity.179.DHS 1. 0: Anterior DHS approximately as straight and as wide as other hemal spines. 1: Anterior DHS large, two to three times as broad as other hemal spines, often exhibiting additional distal tips. In the derived state the anterior DHS is curved and scythe shaped.180.DHS 1 proximal surface. 0: Proximal surface of first DHS narrower than descending blade. 1: Proximal surface of first DHS broad as blade.181.DHS 2 shape. 0: Second posterior DHS straight. 1: Second posterior DHS curved 182.Number posterior DHS. 0: Two or three DHSs posterior to large anterior spine. 1: A single DHS posterior to large anterior spine.183.Dorsal organ. 0: Posterodorsal margin of body without a longitudinal fleshy organ. 1: Posterodorsal margin of body with a median flap or bar of fleshy tissue, extending parallel to the dorsal margin of epaxial musculature 184.Dorsal organ length. 0: Dorsal organ extending along dorsal margin posterior to midlength of body. 1: Dorsal organ extends along entire dorsal margin of body, from nape to caudal peduncle length. 0: Anal-fin pterygiophores shorter than hemal spines at midbody; less than one-third total body depth . 1: Anal-fin pterygiophores longer than hemal spines at midbody; more than one-third total body depth .191.Shape of AFP blades. 0: Descending blades of proximal anal-fin pterygiophores slender, approximately cylindrical in cross section. 1: Descending blades of anal-fin pterygiophores broad, anterior and posterior margins extending into ventral median septum in cross section.192.Shape of AFP tips. 0: Anal-fin pterygiophores tapering smoothly to tips. 1: Tips of pterygiophores shaped like an arrow-head; axial series of pterygiophores providing the ventral margin of the anal-fin base a scalloped appearance 193.Anal-fin ray articulation. 0: Anal-fin rays articulate with distal anal-fin pterygiophores. 1: Anal-fin rays articulate with proximal anal-fin pterygiophores . 1: Body cavity associated with 23\u201329 precaudal vertebrae. 2: Body cavity associated with 30\u201339 precaudal vertebrae. 3: Body cavity associated with 40 or more precaudal vertebrae.197.Body cavity short. 0: Body cavity associated with 16\u201319 vertebrae. 1: Body cavity short; associated with 12\u201315 precaudal vertebrae. 2: Body cavity very short; associated with 11 or fewer precaudal vertebrae vertebrae bearing hemal spines present. 1: Hemal spines absent, body cavity extending almost to tip of the tail; no caudal (post-coelomic) vertebrae.199.Number of pleural ribs. 0: Eight or more pairs of pleural ribs. 1: Seven or fewer pairs of pleural ribs . 0: All axial muscle fibers unmodified; no organs capable of generating rhythmic electric discharges. 1: Paired electrogenic organs developing in larval hypaxial musculature; electric organ composed of rows of modified elongate myofibrils .216.Main EO electrocyte morphology. 0: Electrocytes cigar shaped, elongate; longitudinal axis parallel with neuraxis. 1: Electrocytes barrel shaped, cylindrical; long axis oriented vertically. 2: Electrocytes coin-shaped (new character state).217.Hypaxial EO ontogeny. 0: Main electric organ of mature specimens developing from a medial portion of hypaxial musculature, extending along ventral margin of hypaxial musculature. 1: Hypaxial electric organ replaced during development, adult organ not derived from hypaxial musculature 218.Mental accessory EO. 0: absent. 1: present 219.Mental accessory EO configuration. 0: Mental accessory organ absent or short with few electrocytes. 1: Mental accessory EO long, threadlike with many electrocytes 220.Humeral accessory EO. 0: No humeral electric organ. 1: Humeral electric organ extending dorsally from pectoral fin base, and then posteriorly along horizontal myoseptum a distance less than length of pectoral fin 221.Neural EO. 0: Main electric organ of mature specimens ontogenetically derived from hypaxial musculature. 1: Main electric organ of mature specimens derived from electromotor neurons which innervate larval hypaxial organ 222.EOD form. 0: EOD of mature specimens produced as discrete non-overlapping pulses with alternating periods of current flow and no current flow; capacity for EOD frequency modulations present; cells of pacemaker nucleus organized into two separate clusters. 1: EOD produced as a continual series of discharges to form a quasi-sinusoidal pattern of current emission; no capacity for EOD frequency modulations; relay and pacemaker cells mingled in a single medullary nucleus 223.EOD monophasic in adults. 0: EOD of mature specimens with two (sometimes three or four) phases; EOD characterized by both head-positive and head-negative depolarizations. 1: Monophasic EOD of juveniles retained into maturity; EOD characterized exclusively by head-positive depolarizations. 2: Monophasic hyperpolarization from negative baseline Mkv model MBTE-ML analyses datasets were conducted in Mkv model 3). Each run was comprised of 5.0\u00d7107 generations with model parameter values and a single tree sampled every 5\u00d7103 generation. All other parameters were set as default. To ensure adequate mixing of the MCMC, effective sample size values (ESS>200) were inspected for parameter estimates in Tracer 1.5. The two independent runs were summarized with \u201csump\u201d and \u201csumt\u201d commands in MrBayes 3.2 MBTE-BI analyses were conducted in MrBayes 3.2 Node 175: GYMNOTIFORMESch. 3-Body shape profile. \u201cBody Depth,\u201d character 2 in Albert, 2001. / 1: Body elongate, slender, depth less than 11% total length.ch. 7-Gape short. / 1: Rictus extends to a vertical with mental symphysis, gape very small, less than twice diameter of eye, oriented oblique to long axis of head.ch. 14-Position of eye. / 1: Eye completely covered by epidermis in adults; orbital margin not free.ch. 72-Lateral ethmoid size. / 1: Lateral ethmoid reduced in size; four peripheral margins not contacting other bony surfaces.ch. 84-Parasphenoid lateral process. / 1: Lateral margins of parasphenoid not extending to a horizontal with trigeminal foramen.ch. 106-Accessory optic system. / 1: Accessory optic tract reduced or absent; discrete accessory optic nucleus not visible in sections.ch. 107-Integumental taste buds. / 1: Taste buds entirely absent from extra-oral integument; nerves V and VII smaller than other cranial nerves of isthmal region; primary facial and vagal sensory nuclei smaller than medial octaval nucleus.ch. 108-Schreckstoff/club cells. / 1: Schreckstoff, club cells, and fright response absent.ch. 110-Ampullary organ rosettes. / 1: Ampullary organs clustered in rosettes.ch. 111-Active electroreception. / 1: Electrogeneration and high frequency electroreception, used in communication and navigation (in addition to predation); neural apparatus for producing and detecting high frequency electric currents.ch. 112-Tuberous electroreceptors. / 1: Two classes of morphologically distinct tuberous electroreceptor organs.ch. 120-Palatines. / 1: Autopalatine unossified, arched.ch. 121-Ectopterygoid. / 1: Ectopterygoid and associated teeth absent.ch. 125-Mesopterygoid dentition. / 1: Few or no teeth on endopterygoid.ch. 127-Metapterygoid shape. / 1: Metapterygoid triangular in lateral view.ch. 141-Gill raker configuration. / 1: Base of gill rakers not mineralized, rakers (when present) not attached to gill arches.ch. 148-Shape of 4th epibranchial. / 1: Fourth epibranchial with an elongate ascending process.ch. 165-Mesocoracoid. / 1: Mesocoracoid not ossified.ch. 169-Pelvic girdle and fin. / 1: No pelvic girdles or fins.ch. 170-Claustrum. / 1: Claustrum absent as an ossified element.ch. 185-Dorsal fin. / 1: Dorsal fin absent.ch. 186-Adipose fin. / 1: Adipose fin absent.ch. 188-Number anal-fin rays. / 2: 160\u2013199 rays.ch. 193-Anal-fin ray articulation. / 1: Anal-fin rays articulate with proximal anal-fin pterygiophores.ch. 194-Distal AFP. / 1: No distal anal-fin pterygiophores.ch. 213-Caudal fin. / 1: Caudal fin absent.ch. 214-Electric organs (EO). / 1: Paired electrogenic organs developing in larval hypaxial musculature; electric organ composed of rows of modified elongate myofibrils.ch. 215-Number of hypaxial EO. / 1: Three anatomically distinct hypaxial electric organs .Node 176: GYMNOTIDAE cladech. 8-Oral opening in adults. / 0: Upper and lower jaws of equal length, oral aperture terminal.ch. 64-Mesethmoid, tip size. / 1: Portion of mesethmoid anterior to ventral ethmoid flexed ventrally in mature specimens; its dorsal surface anterior and posterior to ventral ethmoid at an oblique angle; its ventral surface oblique to dorsal surface. terminal.ch. 74-Base lateral ethmoid. / 1: Lateral ethmoid narrow or tubular; length of its base less than one-third length of its anterior margin.ch. 79-Cranial fontanels. / 1: Frontals in contact with each other along the entire extent of their medial margins in mature specimens.ch. 140-Branchiostegal ray morphology. / 0: Anterior 1\u20132 rays broad.ch. 163-Posttemporal. / 0: Posttemporal independent from supracleithrum in mature specimens.ch. 196-Body cavity long. / 2: Body cavity associated with 30\u201339 precaudal vertebrae.ch. 204-Post. chamber gas bladder. / 1: Posterior chamber of gas bladder elongate, passing between hemal arches of postcoelomic axial skeleton and musculature.ch. 210-Tail length. / 1: Tail short, 0\u201316% total length.Gymnotus cladeNode 177: ch. 8-Oral opening in adults. / 1: Lower jaw extends anterior to upper, oral aperture superior.ch. 10-Position of anterior nares. / 1: Anterior nares located very close to or within gape, narial opening oriented anteroventrally.ch. 11-Anterior narial pore. / 1: Anterior narial opening sessile, its rim flush with surrounding integument.ch. 33-Lateral line pores. / 1: Posterior lateral line pores tubular; tube length more than three times pore diameter.ch. 34-Lateral line ventral rami. / 1: Numerous ventral rami extending parallel with lateral line.ch. 37-Tooth shape. / 1: Teeth in both jaws villiform, each tooth a long cylindrical shaft with a narrow base.ch. 47-Maxilla descending blade. / 2: Anteroventral margin of descending blade not ossified; distal half of blade extending as a narrow process with a sharp point at its distal tip.ch. 66-Ventral ethmoid lateral process. / 1: No lateral process of ventral ethmoid; ventral ethmoid not contacting lateral ethmoid cartilage.ch. 67-Ventral ethmoid lateral process shape. / 1: Lateral process of ventral ethmoid robust, posterior surface forming articulation with lateral ethmoid cartilage broad and rounded, covered by a cartilage cap.ch. 68-Median septum of ventral ethmoid. / 1: Ossified median septum of ventral ethmoid elongate in mature specimens, longer than deep, extending posterior to posterior margin of lateral process.ch. 71-Ethmoid cartilage. / 1: Ethmoid cartilage deeper than long; antorbital region of snout about as deep as long.ch. 87-Parasphenoid process. / 1: Parasphenoid with anteroventral process.ch. 91-Nasal loop. / 1: Commissure between infraorbital and supraorbital canals extended anteriorly, forming a loop ventrolateral to nasal capsule; antorbital and first infraorbital bones situated near anterior nares.ch. 92-Infraorbital subnasal extension. / 1: Anterior extension of infraorbital canal shorter than width of canal pore; anterior canal pore of infraorbital canal situated near first infraorbital.ch. 122-Endopterygoid ascending process. / 1: Ascending process on lateral surface of endopterygoid; pterygocranial ligament ossified; base of ascending process situated approximately dorsal to articulation of quadrate with anguloarticular.ch. 143-Anterior pharyngobranchial. / 1: Anterior pharyngobranchial unossified.ch. 158-Basibranchials. / 1: All five elements of basibranchial series unossified.ch. 195-Free neural and hemal spines. / 1: Capacity to generate series of free neural and hemal spines associated with regenerated cartilaginous rod.ch. 201-Size of anterior ribs. / 1: Anterior ribs broad, breadth two to three times width.Gymnotus pantherinus cladeNode 178: No diagnostic character in matrix.Gymnotus coatesi+G. anguillaris+G. tigre+G. cylindricus+G. carapo cladesNode 180: ch. 16-Oblique pigment bands. / 1: Multiple (13\u201350) pale bands with straight margins of alternating high and low melanophore density along lateral surface of body, oriented at an oblique angle to longitudinal body axis. Bands sometimes interrupted by patches of depigmented integument, resulting in a distribution of blotches arranged in oblique bands along the lateral surface of body.ch. 35-No. ventral rami. / 1: Median 15 or more.Gymnotus coatesi cladeNode 181: No diagnostic character in matrix. See Maxime (2014).Gymnotus anguillaris+G. tigre+G. cylindricus+G. carapo cladeNode 187: ch. 188-Number anal-fin rays. / 3: 200\u2013299 rays. Taxa coded by modal number of anal-fin rays.ch. 196-Body cavity long. / 3: Body cavity associated with 40 or more precaudal vertebrae.Gymnotus cataniapo cladeNode 188: No diagnostic character in matrix.Gymnotus tigre+G. cylindricus species groups+G. carapo cladeNode 190: ch. 16-Oblique pigment bands. / 2: Oblique pigment bands along longitudinal axis with wavy margins.ch. 37-Tooth shape. / 0: Teeth in both jaws conical, with a broad base tapering toward the cusp.ch. 135-Preopercular pores. / 1: Two pores at dorsoposterior corner of preopercle.ch. 165-Mesocoracoid. / 0: Mesocoracoid ossified within scapulocoracoid cartilage, forming a bridge between medial surface of coracoid and cleithrum.ch. 190-Anal-fin pterygiophore (AFP) length. / 1: Anal-fin pterygiophores longer than hemal spines at midbody; more than one-third total body depth .ch. 201-Size of anterior ribs. / 0: Anterior pair of pleural ribs narrow; breadth approximately equal to width.Gymnotus tigre cladeNode 191: ch. 15-Anal fin membrane. / 2: Anal fin membrane striped.ch. 38-Tooth tip shape. / 1: Tips of teeth directed anteriorly (recurved).Gymnotus cylindricus+G. carapo cladeNode 192: ch. 196-Body cavity long. / ch.196-Body cavity long. / 2: Body cavity associated with 30\u201339 precaudal vertebrae.Gymnotus cylindricus cladeNode 193: ch. 16-Oblique pigment bands. / 0: Body pigmentation evenly distributed along longitudinal axis.ch. 35-No. ventral rami. / 0: Median 14 or less.ch. 135-Preopercular pores. / 0: One pore at dorsoposterior corner of preoperculum.ch. 188-Number anal-fin rays. / 2: 160\u2013199 rays.ch. 190-Anal-fin pterygiophore (AFP) length. / 0: Anal-fin pterygiophores shorter than hemal spines at midbody; less than one-third total body depth .G. carapo cladeNode 196: ch. 15-Anal fin membrane. / 1: With pale posterior patch.ch. 37-Tooth shape. / 2: Teeth with triangular, arrow-head shape.Gymnotus carapo species-complex cladeNode 201: ch. 1-Body shape 1. / 0: Body laterally compressed, body width at pectoral fin base less than 70% its depth.ch. 3-Body shape profile. / 0: Body relatively deep in profile, depth at pectoral girdle more than 11% total length.ch. 157-Basihyal dorsal groove. / 1: Dorsal surface of basihyal concave along its long axis, forming a shallow trough.ch. 166-Anterior coracoid process. / 1: Anterior coracoid process not extending to a vertical with contact of dorsomedial limb of coracoid with cleithrum.Node 216: STERNOPYGOIDEI cladech. 1-Body shape 1. / 0: Body laterally compressed, body width at pectoral fin base less than 70% its depth.ch. 27-Branchial opening. / 1: Vertical extent of branchial opening restricted to region around pectoral fin base; ventral portion reduced by a dorsolateral continuation of epidermis from isthmus.ch. 28-Pseudotympanum. / 1: Sixth vertebra not covered by superior oblique.ch. 39-Premaxilla size. / 1: Small. The anterodorsal orientation of the articular surface of the maxilla with the autopalatine is a consequence of the large size of the premaxilla and the associated posterior position of the maxilla.ch. 62-Mesethmoid, tip size. / 1: Anterior tip of mesethmoid small, anterodorsal surface narrow, with a median knob-shaped process directed anteriorly between two small lateral processes.ch. 63-Mesethmoid, tip shape. / 1: Portion of mesethmoid anterior to ventral ethmoid flexed ventrally in mature specimens; its dorsal surface anterior and posterior to ventral ethmoid at an oblique angle; its ventral surface oblique to dorsal surface.ch. 115-Lateral line afferents. / 1: Lateral line afferents fasciculated into discrete bundles; fibers from each lateral line nerve segregated from those of other lateral line nerves.ch. 139-Branchiostegal rays. / 1: 5\u20136 rays.ch. 160-Urohyal head. / 1: Anterior head of urohyal large, with lateral ridges.ch. 177-Displaced hemal spines (DHS). / 1: Three additional hemal spines positioned in hypaxial musculature posterior to body cavity, often lateral to unmodified hemal spines, rarely fused with hemal arches or parapophyses; irregular association with posterior thoracic and anterior caudal vertebrae.ch. 187-Anal fin origin. / 1: Anal-fin origin ventral to posterior margin of cleithrum.ch. 197-Body cavity short. / 1: Body cavity short; associated with 12\u201315 precaudal vertebrae.ch. 206-Anal position. / 1: Position of anus changing allometrically during ontogeny, starting near posterior end of coelomic cavity and growing anterior to pectoral girdle; anus located near isthmus.ch. 211-Elongate caudal rod. / 1: cartilaginous bar or rod, regenerated in place of caudal vertebrae.Node 217: RHAMPHICHTHYOIDEA cladech. 36-Adult dentition. / 1: Oral teeth present in juveniles, lost and not replaced during development.ch. 66-Ventral ethmoid lateral process. / 1: No lateral process of ventral ethmoid; ventral ethmoid not contacting lateral ethmoid cartilage.ch. 81-Orbitosphenoid shape. / 1: Anterior margin of orbitosphenoid not ossified, orbitosphenoid narrow, its ventral margin about as long or shorter than its dorsal margin.ch. 87-Parasphenoid process. / 1: Parasphenoid with anteroventral process.ch. 95-Antorbital size. / 1: Antorbital large; its ventral portion larger than maxilla; expanded dorsal portion contacts autopalatine.ch. 98-Infraorbital canal tube. / 1: Infraorbital canal a single, lightly ossified continuous tube.ch. 99-First infraorbital. / 1: First infraorbital not present as a separate ossification.ch. 118-Anterior corpus cerebellum. / 0: Anterior lobe of corpus cerebellum large, extending anterior to midlength of optic tectum; cerebellum overlying commissure of optic tectum.ch. 128-Metapterygoid posterior wing. / 1: Metapterygoid elongate and narrow, longer than wide at its midlength.ch. 134-Preopercular orientation. / 1: Long axis of preopercle horizontal, roughly parallel with main axis of neurocranium.ch. 137-Shape of opercle. / 0: Outline of opercle approximately rectangular; dorsal margin shorter than posterior margin, and interrupted by a pronounced angle.ch. 181-DHS 2 shape. / 1: Second posterior DHS curved.ch. 208-Urogenital papilla. / 1: Urogenital pore elevated onto a papilla in sexually mature specimens.Akawaio, Hypopomus, Microsternarchini, Brachyhypopomus) cladeNode 218: Hypopomidae cladeNode 221: Microsternarchini ch. 140-Branchiostegal ray morphology. / 0: Anterior 1\u20132 rays broad.ch. 158-Basibranchials. / 1: All five elements of basibranchial series unossified.Procerusternarchus+MicrosternarchusNode 222: ch. 139-Branchiostegal rays. / 0: 3\u20134 rays.Brachyhypopomus cladeNode 224: ch. 8-Oral opening in adults. / 0: Upper and lower jaws of equal length, oral aperture terminal.ch. 74-Base lateral ethmoid. / 1: Lateral ethmoid narrow or tubular; length of its base less than one-third length of its anterior margin.ch. 171-Os suspensorium. / 1: Anterior ramus short, reaching second vertebra.ch. 188-Number anal-fin rays. / 3: 200\u2013299 rays. Taxa coded by modal number of anal-fin rays.ch. 212-Caudal appendage. / 1: Caudal appendage elongate in sexually mature males.Node 234: RHAMPHICHTHYIDAE cladech. 210-Tail length. / 2: Tail long, more than 45% total length.ch. 218-Mental accessory EO. / 1: present.Steatogenys, Hypopygus) cladeNode 235: Steatogenae , a ventral extension of the medial surface of dentary where it covers the anterior portion of Meckel\u05f3s cartilage.ch. 55-Dentary teeth size. / 1: Teeth on posterior half of dentary twice the size of anterior teeth.ch. 71-Ethmoid cartilage. / 1: Ethmoid cartilage deeper than long; antorbital region of snout about as deep as long.ch. 73-Lateral ethmoid. / 1: Lateral ethmoid not ossified.ch. 86-Parasphenoid dorsal margin. / 1: Dorsal margin narrow.ch. 140-Branchiostegal ray morphology. / 0: Anterior 1\u20132 rays broad.ch. 143-Anterior pharyngobranchial. / 1: Anterior pharyngobranchial unossified.ch. 149-Epibranchial 5. / 1: Fifth epibranchial with posterior process.ch. 153-Hypobranchial 1. / 2: First hypobranchial rounded or pentagonal in dorsal view; anterior margin interrupted by a sharp angle.ch. 162-Urohyal blade hyperossified. / 1: Urohyal blade long, ossified to level of fourth basibranchial.ch. 188-Number anal-fin rays. / 1: Anal fin long, extending along majority of ventral body margin; 100\u2013159 rays.ch. 197-Body cavity short. / 2: Body cavity very short; associated with 11 or fewer precaudal vertebrae.ch. 208-Urogenital papilla. / 0: Urogenital pore sessile, opening flush with ventral margin of body wall in sexually mature specimens.ch. 220-Humeral accessory EO. / 1: Humeral electric organ extending dorsally from pectoral fin base, and then posteriorly along horizontal myoseptum a distance less than length of pectoral fin.Steatogenys cladeNode 238: ch. 18-Vertical pigment bars. \u201cSaddle-shaped bars\u201d. / 1\u201310 dark bars across mid-dorsal surface extending as vertical bands onto lateral surfaces.ch. 165-Mesocoracoid. / 0: Mesocoracoid ossified within scapulocoracoid cartilage, forming a bridge between medial surface of coracoid and cleithrum.ch. 166-Anterior coracoid process. / 1: Anterior coracoid process not extending to a vertical with contact of dorsomedial limb of coracoid with cleithrum.ch. 167-Proximal pectoral radials. / 1: Proximal radials three and four co-ossified in adult specimens.ch. 219-Mental accessory EO configuration. / 1: Mental accessory EO long, threadlike with many electrocytes.Hypopygus cladeNode 236: ch. 10-Position of anterior nares. / 1: Anterior nares located very close to or within gape, narial opening oriented anteroventrally.ch. 12-Posterior narial pore. / 1: Posterior narial pore absent.ch. 17-Vertical pigment lines. / 1: Thin vertical pigment lines present along longitudinal body axis.ch. 32-Lateral line. / 1: Lateral line incomplete.ch. 70-Dermal vomer. / 1: Dermal vomer not ossified.ch. 163-Posttemporal. / 0: Posttemporal independent from supracleithrum in mature specimens.Node 240: Rhamphichthyinae cladech. 5-Snout long. / 1: Snout elongate, frontal, vomer and anterior portion of parasphenoid elongate; preorbital length longer than one-third total head length or greater in mature specimens.ch. 9-Position of nasal capsule. / 1: Anterior position of nasal capsule; located closer to tip of snout than to eye; posterior nares closer to anterior nares than to anterior margin of eye.ch. 30-Scales on middorsum. / 1: Scales absent from head, anterior portion of dorsal midline, and area dorsal to pectoral fins.ch. 33-Lateral line pores. / 1: Posterior lateral line pores tubular; tube length more than three times pore diameter.ch. 59-Posterior limb anguloarticular. / 1: Posterior limb of anguloarticular large; its ventral margin longer than that of retroarticular.ch. 61-Mesethmoid length. / 1: Mesethmoid elongate, its length greater than antorbital region of frontal.ch. 87-Parasphenoid process. / 0: Anteroventral margin of parasphenoid smooth.ch. 116-Anterior extent of eminentia granularis. / 1: EG small, its anterior margin not extending to contact optic tectum.ch. 124-Endopterygoid anterior process. / 1: Entire extent of ligament ossified, forming a bony strut anterior to orbit; process equally as wide along most of its length.ch. 129-Size of symplectic. / 1: Length of symplectic greater than hyomandibula.ch. 130-Orientation of hyomandibula. / 1: Main axis of hyomandibula oriented horizontally, parallel to main axis of neurocranium.ch. 138-Opercular dorsal margin. / 1: Dorsal margin of opercle straight.ch. 178-DHS anterior series. / 1: Anterior series of 8\u201314 DHSs in hypaxial musculature lateral to body cavity.ch. 181-DHS 2 shape. / 0: Second posterior DHS straight.ch. 187-Anal fin origin. / 2: Anal fin origin near branchial isthmus.ch. 189-Anal-fin rays unbranched. / 2: 30\u201360 unbranched anal-fin rays.ch. 190-Anal-fin pterygiophore (AFP) length. / 1: Anal-fin pterygiophores longer than hemal spines at midbody; more than one-third total body depth .Gymnorhamphichthys cladeNode 241: ch. 30-Scales on middorsum. / 2: Scales absent along entire middorsum.ch. 43-Anterior maxillary process. / 1: Anterior process of maxilla extends anterior to articulation of maxilla and autopalatine, forming a tapered process, its ventral margin continuous with descending blade of maxilla; maxilla forked in lateral view.ch. 51-Dentary dorsal margin. / 1: Dorsal margin of dentary concave.ch. 69-Ventral ethmoid-Vomer. / 1: Ventral ethmoid and vomer not fused in adultsch. 82-Orbitosphenoid margin. \u201cSphenoid fenestra\u201d / 1: Posterior margin of orbitosphenoid not contacting pterosphenoid, except between dorsal portion of their common margin; presence of an unmineralized fenestra between orbitosphenoid and pterosphenoid.Iracema, Rhamphichthys) cladeNode 249: Rhamphichthyini .ch. 88-Posttemporal fossa. / 1: Epioccipital, pterotic, and parietal bones not contacting one another along their mutual margins, forming a fossa in posttemporal region; lateral surface of otic capsule exposed.ch. 94-Antorbital. / 1: Infraorbital canal extending onto antorbital.ch. 117-Posterior EG. / 1: Posterior lobe of EG well developed, wrapped around caudal lobe of cerebellum, its posterior margin extending to a vertical with posterior margin of ELL.ch. 122-Endopterygoid ascending process. / 1: Ascending process on lateral surface of endopterygoid; pterygocranial ligament ossified; base of ascending process situated approximately dorsal to articulation of quadrate with anguloarticular.ch. 146-Epibranchial 4. / 1: Fourth epibranchial with short posterior process.ch. 173-Position of neural spines. / 1: Neural spine inserting on posterior margin of caudal vertebral centra.ch. 174-Vertebral fenestrae. / 1: Lateral walls of neural arches with several small fenestrae; dorsal margin uneven, with several evaginations.ch. 179-DHS 1. / 1: Anterior DHS large, two to three times as broad as other hemal spines, often exhibiting additional distal tips. In the derived state the anterior DHS is curved and scythe shaped.ch. 180-DHS 1. / 1: Proximal surface of first DHS broad as blade.ch. 188-Number anal-fin rays. / 3: 200\u2013299 rays. Taxa coded by modal number of anal-fin rays.ch. 189-Anal-fin rays unbranched. / 1: Anterior 15\u201325 rays anal-fin rays unbranched to their tips.ch. 194-Distal AFP. / 0: Distal anal-fin pterygiophores present.ch. 217-Hypaxial EO ontogeny. / 1: Hypaxial electric organ replaced during development, adult organ not derived from hypaxial musculature.ch. 222-EOD form. / 1: EOD produced as a continual series of discharges to form a quasi-sinusoidal pattern of current emission; no capacity for EOD frequency modulations; relay and pacemaker cells mingled in a single medullary nucleus.ch. 223-EOD monophasic in adults. / 1: Monophasic EOD of juveniles retained into maturity; EOD characterized exclusively by head-positive depolarizations.Node 260: STERNOPYGIDAE cladech. 8-Oral opening in adults. / 0: Upper and lower jaws of equal length, oral aperture terminal.ch. 13-Eye size. / 0: Eye and optic tract large; about two eye diameters into postorbital head length.ch. 37-Tooth shape. / 1: Teeth in both jaws villiform, each tooth a long cylindrical shaft with a narrow base.ch. 57-M. Adductor mandibulae. / 1: Additional insertion of A1 on first infraorbital.ch. 72-Lateral ethmoid size. / 0: Lateral ethmoid a large endochondral ossification in the antorbital region, arching laterally over Profundus (V1) nerve, with four margins; anterolateral process contacting ventral ethmoid, posteromedial process contacting parasphenoid, dorsomedial margin contacting frontal, and anteromedial margin contacting mesethmoid.ch. 75-Nasal. / 1: Nasal broad.ch. 77-Antorbital process frontals. / 1: Lateroventral process of frontals anterior to orbit.ch. 97-Infraorbital canal plates. / 1: Antorbital and infraorbitals 1\u20134 large, partial cylinders with slender osseous arches.ch. 100-Mandibular canal. / 0: Canal bearing bones of preopercular\u2013mandibular laterosensory canal long and slender ossifications embedded in dermis; diameter of canal slender.ch. 106-Accessory optic system. / 0: Accessory optic tract large, easily visible in histological sections; neurons of tract organized into a distinct tegmental cell cluster .ch. 133-Mandibular canal size. / 1: Mandibular canal ossicles dumbbell-shaped.ch. 152-Ceratobranchial 4. \u201cLateral process of sixth ceratobranchial\u201d. / 1: Lateral surface of fourth ceratobranchial with an anterolaterally oriented process.ch. 172-Anterior vertebrae. / 1: Parapophyses of second vertebrae separated by distinct gap from the os suspensorium.ch. 176-Caudal intermusculars. / 1: Capacity to regenerate ossified intermuscular bones.ch. 197-Body cavity short. / 0: Body cavity associated with 16\u201319 vertebrae.Sternopygus cladeNode 261: ch. 14-Position of eye. / 0: Surface of eye not covered by epidermis in adults; free orbital margin.ch. 20-Longitudinal lines. / 2: A white narrow stripe extending parallel to the base of the anal-fin pterygiophores, and then posteriorly along the lateral midline.ch. 27-Branchial opening. / 0: Branchial opening extends along entire posterior margin of opercle, from isthmus to pectoral fin insertion.ch. 63-Mesethmoid, tip shape. / 0: Portion of mesethmoid anterior to ventral ethmoid horizontal; its dorsal surface anterior and posterior to ventral ethmoid approximately parallel; its ventral surface parallel with dorsal surface.ch. 68-Median septum of ventral ethmoid. / 1: Ossified median septum of ventral ethmoid elongate in mature specimens, longer than deep, extending posterior to posterior margin of lateral process.ch. 125-Mesopterygoid dentition. / 0: Numerous small teeth distributed in an irregular field on anterior portion of ventral surface of endopterygoid.ch. 163-Posttemporal. / 0: Posttemporal independent from supracleithrum in mature specimens.ch. 189-Anal-fin rays unbranched. / 3: all anal-fin rays unbranched.ch. 216-Main EO electrocyte morphology. / 0: Electrocytes cigar shaped, elongate; longitudinal axis parallel with neuraxis.Node 265: Eigenmanninae cladech. 4-Snout length short. / 1: Snout short, preorbital length less than one-third total head length.ch. 26-Body translucence. / 1: Body translucent in living specimens, yellow or pink hue in living specimens, yellow or hyaline in formalin-fixed specimens, melanophores sparse or absent on lateral body surface.ch. 83-Sphenotic process. / 1: Dorsolateral margin of sphenotic bearing a transversely oriented crest or process exposed on dorsolateral edge, anterior margin not underlying frontal.ch. 146-Epibranchial 4. / 0: Posterior margin of fourth epibranchial flat.ch. 148-Shape of 4th epibranchial. / 0: Fourth epibranchial with short ascending process.ch. 164-Scapular foramen. / 1: Unossified region of scapulocoracoid cartilage included entirely within the scapula, forming a large foramen.ch. 167-Proximal pectoral radials. / 1: Proximal radials three and four co-ossified in adult specimens.ch. 175-Shape anterior intermusculars. / 1: Intermusculars highly branched.ch. 180-DHS 1 proximal surface. / 0: Proximal surface of first DHS narrower than descending blade.ch. 188-Number anal-fin rays. / 2: 160\u2013199 rays.ch. 197-Body cavity short. / 2: Body cavity very short; associated with 11 or fewer precaudal vertebrae.ch. 199-Number of pleural ribs. / 1: Seven or fewer pairs of pleural ribs.ch. 200-Length of anterior ribs. / 1: Length of anterior two ribs greater than 80% body depth at pectoral girdle.Rhabdolichops cladeNode 266: ch. 30-Scales on middorsum. / 1: Scales absent from head, anterior portion of dorsal midline, and area dorsal to pectoral fins.ch. 43-Anterior maxillary process. / 0: Anterior process of maxilla absent.ch. 46-Maxilla descending blade. / 1: Descending blade of maxilla broad, connective tissue membrane along its anteroventral margin ossified to form a thin shelf; anterior portion of maxilla rhomboid in lateral view.ch. 86-Parasphenoid dorsal margin. / 1: Dorsal margin narrow.ch. 87-Parasphenoid process. / 1: Parasphenoid with anteroventral process.ch. 182-Number posterior DHS. / 1: A single DHS posterior to large anterior spine.ch. 190-Anal-fin pterygiophore (AFP) length. / 1: Anal-fin pterygiophores longer than hemal spines at midbody; more than one-third total body depth .Distocyclus, Archolaemus, Japigny, Eigenmannia) cladeNode 270: Eigenmannini length. / 1: Anal-fin pterygiophores longer than hemal spines at midbody; more than one-third total body depth .ch. 202-Posterior parapophyses. / 1: Parapophyses of posterior precaudal vertebra longer than wide, their ventral margins parallel with long axis of body, abutting at midline.ch. 203-Shape last precaudal parapophyses. / 1: Parapophyses of last precaudal vertebra slender and sinuous, their tips pointed.ch. 210-Tail length. / 1: Tail short, 0\u201316% total length.ch. 211-Elongate caudal rod. / 0: Caudal fin present with hypural plate and segmented rays.ch. 213-Caudal fin. / 0: Caudal fin present.ch. 221-Neural EO. / 1: Main electric organ of mature specimens derived from electromotor neurons which innervate larval hypaxial organ.Orthosternarchus, Sternarchorhamphus) cladeNode 280: Sternarchorhamphinae .ch. 123-Endopterygoid ascending process. / 1: Small ascending process of endopterygoid in juveniles obliterated by growth along dorsal margin of bone; no endopterygoid process in adults.ch. 128-Metapterygoid posterior wing. / 1: Metapterygoid elongate and narrow, longer than wide at its midlength.ch. 165-Mesocoracoid. / 0: Mesocoracoid ossified within scapulocoracoid cartilage, forming a bridge between medial surface of coracoid and cleithrum.ch. 184-Dorsal organ length. / 1: Dorsal organ extends along entire dorsal margin of body, from nape to caudal peduncle.ch. 187-Anal fin origin. / 2: Anal fin origin near branchial isthmus.ch. 189-Anal-fin rays unbranched. / 3: all anal-fin rays unbranched.ch. 191-Shape of AFP blades. / 1: Descending blades of anal-fin pterygiophores broad, anterior and posterior margins extending into ventral median septum in cross section.ch. 192-Shape of AFP tips. / 1: Tips of pterygiophores shaped like an arrow-head; axial series of pterygiophores providing the ventral margin of the anal-fin base a scalloped appearance.Node 281: Apteronotinae cladech. 67-Ventral ethmoid lateral process shape. / 1: Lateral process of ventral ethmoid robust, posterior surface forming articulation with lateral ethmoid cartilage broad and rounded, covered by a cartilage cap.ch. 153-Hypobranchial 1. / 1: First hypobranchial triangular in dorsal view. 2: First hypobranchial rounded or pentagonal in dorsal view; anterior margin interrupted by a sharp angle.ch. 188-Number anal-fin rays. / 1: Anal fin long, extending along majority of ventral body margin; 100\u2013159 rays.ch. 201-Size of anterior ribs. / 1: Anterior ribs broad, breadth two to three times width.ch. 223-EOD monophasic in adults. / 0: EOD of mature specimens with two (sometimes three or four) phases; EOD characterized by both head-positive and head-negative depolarizations.Adontosternarchus cladeNode 282: ch. 4-Snout length short. / 1: Snout short, preorbital length less than one-third total head length.ch. 8-Oral opening in adults. / 0: Upper and lower jaws of equal length, oral aperture terminal.ch. 36-Adult dentition. / 1: Oral teeth present in juveniles, lost and not replaced during development.ch. 40-Snout length short. / 1: Snout short, preorbital length less than one-third total head length.ch. 47-Maxilla descending blade. / 2: Anteroventral margin of descending blade not ossified; distal half of blade extending as a narrow process with a sharp point at its distal tip.ch. 50-Dentary gracile. / 1: Dentary gracile, posterodorsal process tapering to a point (except in Adontosternarchus sachsi), ventral margin concave.ch. 71-Ethmoid cartilage. / 1: Ethmoid cartilage deeper than long; antorbital region of snout about as deep as long.ch. 81-Orbitosphenoid shape. / 1: Anterior margin of orbitosphenoid not ossified, orbitosphenoid narrow, its ventral margin about as long or shorter than its dorsal margin.ch. 85-Parasphenoid ventral margin. / 1: Ventral margin of parasphenoid flexed sharply on either side of the basicranial region; ventral margin of sphenoid region oblique relative to long axis of neurocranium.ch. 115-Lateral line afferents. / 0: Lateral line afferents from electrosensory periphery intermingled as they course into the electrosensory lateral line lobe (ELL); fibers from different lateral line nerves not segregated.ch. 138-Opercular dorsal margin. / 0: Dorsal margin of opercle convex. 1: Dorsal margin of opercle straight.ch. 139-Branchiostegal rays. / 1: 5\u20136 rays.ch. 161-Urohyal blade. / 1: Posterior blade of urohyal unossified, anterior head of urohyal positioned ventral to second basibranchial.ch. 202-Posterior parapophyses. / 0: Parapophyses of posterior precaudal vertebra small, their ventral margins oblique to long axis of body, not contacting one another along midline.Node 286: Apteronotini+Sternarchorhynchini+Navajini cladech. 64-Mesethmoid, tip size. / 1: Portion of mesethmoid anterior to ventral ethmoid flexed ventrally in mature specimens; its dorsal surface anterior and posterior to ventral ethmoid at an oblique angle; its ventral surface oblique to dorsal surface. terminal.ch. 92-Infraorbital subnasal extension. / 0: Anterior portion of infraorbital canal extending anterior from first infraorbital ventral to nasal capsule; anterior canal pore of infraorbital canal situated anterior to first infraorbital.ch. 203-Shape last precaudal parapophyses. / 0: Parapophyses of last precaudal vertebra broad and triangular, their tips rounded.Parapteronotus, Megadontognathus, Apteronotus) cladeNode 287: Apteronotini length. / 0: Anal-fin pterygiophores shorter than hemal spines at midbody; less than one-third total body depth .Megadontognathus+Apteronotus cladeNode 289: ch. 21-Pigment contrast. / 1: High contrast dark brown or black and white pigments on body surface.ch. 58-Anterior limb anguloarticular. / 1: Anterior limb of anguloarticular shorter than posterior limb.ch. 90-Cranial skeleton texture. / 0: Surface of endochondral and dermal ossifications of cranial skeleton composed of lamellar or cancellous bone.ch. 197-Body cavity short. / 0: Body cavity associated with 16\u201319 vertebrae.ch. 210-Tail length. / 0: Length of tail posterior to anal-fin 17\u201345% total length.Apteronotus cladeNode 290: ch. 19-Caudal Peduncle Spot. / 1: Pale spot present at base of caudal region. Newly coded herein.ch. 23-White mid-sagittal pigments. / 1: Mid-sagittal region of dorsal and mental surfaces bright white.ch. 154-Hypobranchial 2. / 1: Anterior tip of second hypobranchial with a large medially oriented process, contacting contralateral third hypobranchial across midline by means of a cartilaginous bridge.Apteronotus magdalenensis+A. leptorhynchus cladeNode 291: ch. 5-Snout long. / 1: Snout elongate, frontal, vomer and anterior portion of parasphenoid elongate; preorbital length longer than one-third total head length or greater in mature specimens.ch. 80-Sphenoid region. / 1: Sphenoid region of neurocranium more than one-third total head length, combined axial length of the orbitosphenoid and pterosphenoid bones greater than preorbital region.Apteronotus magdalenensis cladeNode 292: ch. 6-Gape large. / 0: Rictus of mouth extends ventral to nasal capsule, gape forming less than one-third total head length.ch. 25-Pigment distribution. / 1: Black and white pigments distributed unevenly over body surface, darker and paler areas grading into one another; integument with a marbled or mottled appearance.ch. 122-Endopterygoid ascending process. / 0: Lateral surface of endopterygoid smooth; no ascending process ossified in pterygocranial ligament (connecting endopterygoid with neurocranium).ch. 123-Endopterygoid ascending process. / 1: Small ascending process of endopterygoid in juveniles obliterated by growth along dorsal margin of bone; no endopterygoid process in adults.Apteronotus leptorhynchus cladeNode 293: ch. 55-Dentary teeth size. / 1: Teeth on posterior half of dentary twice the size of anterior teeth.Apteronotus albifrons cladeNode 294: ch. 22-White posterior bars. / 1: White or pale bars present on caudal region as observed in members of the Apteronotus albifrons species group. Newly coded herein.ch. 197-Body cavity short. / 1: Body cavity short; associated with 12\u201315 precaudal vertebrae.Node 298: Sternarchorhynchini+Navajini cladeNo known diagnostic character.Platyurosternarchus, Sternarchorhynchus) cladeNode 299: Sternarchorhynchini .ch. 123-Endopterygoid ascending process. / 1: Small ascending process of endopterygoid in juveniles obliterated by growth along dorsal margin of bone; no endopterygoid process in adults.ch. 128-Metapterygoid posterior wing. / 1: Metapterygoid elongate and narrow, longer than wide at its midlength.ch. 153-Hypobranchial 1. / 0: First hypobranchial rectangular in dorsal view; anterior margin straight.ch. 187-Anal fin origin. / 2: Anal fin origin near branchial isthmus.ch. 188-Number anal-fin rays. / 2: 160\u2013199 rays.ch. 191-Shape of AFP blades. / 1: Descending blades of anal-fin pterygiophores broad, anterior and posterior margins extending into ventral median septum in cross section.ch. 202-Posterior parapophyses. / 0: Parapophyses of posterior precaudal vertebra small, their ventral margins oblique to long axis of body, not contacting one another along midline.Platyurosternarchus cladeNode 300: ch. 6-Gape large. / 1: Rictus extends posterior to a vertical through eye, gape forming more than one-third total head length.ch. 7-Gape short. / 0: Rictus extends ventral to nasal capsule, gape more than three times eye diameter, oriented parallel with long axis of head.ch. 125-Mesopterygoid dentition. / 0: Numerous small teeth distributed in an irregular field on anterior portion of ventral surface of endopterygoid.ch. 182-Number posterior DHS. / 0: Two or three DHSs posterior to large anterior spine.ch. 188-Number anal-fin rays. / 3: 200\u2013299 rays. Taxa coded by modal number of anal-fin rays.Sternarchorhynchus cladeNode 301: ch. 8-Oral opening in adults. / 0: Upper and lower jaws of equal length, oral aperture terminal.ch. 9-Position of nasal capsule. / 1: Anterior position of nasal capsule; located closer to tip of snout than to eye; posterior nares closer to anterior nares than to anterior margin of eye.ch. 23-White mid-sagittal pigments. / 1: Mid-sagittal region of dorsal and mental surfaces bright white.ch. 45-Anterior maxillary shelf. / 0: Anterior process of maxilla extending as a shelf of bone less than one-third the length of the descending blade.ch. 53-Dentary filamentous. / 1: Dentary elongate and filamentous, more than four times as long as deep.ch. 81-Orbitosphenoid shape. / 1: Anterior margin of orbitosphenoid not ossified, orbitosphenoid narrow, its ventral margin about as long or shorter than its dorsal margin.ch. 94-Antorbital. / 0: Infraorbital canal not extending onto antorbital.ch. 139-Branchiostegal rays. / 1: 5\u20136 rays.ch. 146-Epibranchial 4. / 0: Posterior margin of fourth epibranchial flat.Node 312: Navajini cladech. 26-Body translucence. / 1: Body translucent in living specimens, yellow or pink hue in living specimens, yellow or hyaline in formalin-fixed specimens, melanophores sparse or absent on lateral body surface.ch. 30-Scales on middorsum. / 2: Scales absent along entire middorsum.ch. 31-Scale shape. / 1: Scales dorsal to lateral line rhomboid, their long axis oriented oblique to long axis of body, their dorsoventral axes longer than their longitudinal axes.ch. 49-Rows of dentary teeth. / 1: Teeth on dentary arranged in two to three rows at its midlength.ch. 101-Mandibular canal ossicles. / 1: Canal bearing bones of mandibular laterosensory canal ossified as short, broad, dumbbell-shaped ossicles.Pariosternarchus, Sternarchella, Magosternarchus) cladeNode 313: Sternarchellini nerve, with four margins; anterolateral process contacting ventral ethmoid, posteromedial process contacting parasphenoid, dorsomedial margin contacting frontal, and anteromedial margin contacting mesethmoid.ch. 101-Mandibular canal ossicles. / 0: Canal bearing bones of mandibular laterosensory canal long and slender tubes.ch. 155-Hypobranchial teeth. / 1: Seven or fewer teeth present on sixth hypobranchial.Sternarchogiton, Compsaraia, Porotergus, \"Apteronotus\" bonapartiiNode 316: clade comprised of ch. 19-Caudal Peduncle Spot. / 1: Pale spot present at base of caudal region. Newly coded herein.ch. 46-Maxilla descending blade. / 1: Descending blade of maxilla broad, connective tissue membrane along its anteroventral margin ossified to form a thin shelf; anterior portion of maxilla rhomboid in lateral view.ch. 161-Urohyal blade. / 1: Posterior blade of urohyal unossified, anterior head of urohyal positioned ventral to second basibranchial.ch. 168-Pectoral fin. / 1: Pectoral fin, less than 43% head length.ch. 210-Tail length. / 0: Length of tail posterior to anal-fin 17\u201345% total length.Sternarchogiton cladeNode 317: ch. 4-Snout length short. / 1: Snout short, preorbital length less than one-third total head length .ch. 40-Snout length short. / 1: Snout short, preorbital length less than one-third total head length .ch. 47-Maxilla descending blade. / 1: Ventral margin of descending blade with a sharp angle about two-thirds distance to its tip; ventral margin posterior to this angle relatively straight.ch. 49-Rows of dentary teeth. / 0: A single row of teeth on dentary.ch. 50-Dentary gracile. / 1: Dentary gracile, posterodorsal process tapering to a point (except in Adontosternarchus sachsi), ventral margin concave.ch. 74-Base lateral ethmoid. / 1: Lateral ethmoid narrow or tubular; length of its base less than one-third length of its anterior margin.ch. 81-Orbitosphenoid shape. / 1: Anterior margin of orbitosphenoid not ossified, orbitosphenoid narrow, its ventral margin about as long or shorter than its dorsal margin.ch. 85-Parasphenoid ventral margin. / 1: Ventral margin of parasphenoid flexed sharply on either side of the basicranial region; ventral margin of sphenoid region oblique relative to long axis of neurocranium.ch. 93-Infraorbital\u2013supraorbital prenasal commissure. / 1: Infraorbital\u2013supraorbital prenasal commissure present.ch. 163-Posttemporal. / 0: Posttemporal independent from supracleithrum in mature specimens.ch. 191-Shape of AFP blades. / 1: Descending blades of anal-fin pterygiophores broad, anterior and posterior margins extending into ventral median septum in cross section.ch. 197-Body cavity short. / 2: Body cavity very short; associated with 11 or fewer precaudal vertebrae.Compsaraia+Porotergus gimbeli+\"Apteronotus\" bonapartii species group cladeNode 322: ch. 6-Gape large. / 1: Rictus extends posterior to a vertical through eye, gape forming more than one-third total head length.ch. 7-Gape short. / 0: Rictus extends ventral to nasal capsule, gape more than three times eye diameter, oriented parallel with long axis of head.ch. 9-Position of nasal capsule. / 1: Anterior position of nasal capsule; located closer to tip of snout than to eye; posterior nares closer to anterior nares than to anterior margin of eye.ch. 154-Hypobranchial 2. / 1: Anterior tip of second hypobranchial with a large medially oriented process, contacting contralateral third hypobranchial across midline by means of a cartilaginous bridge.ch. 159-Basibranchial one. / 1: First basibranchial foreshortened and broad, hourglass shaped, breadth at midlength narrower than at either end.Compsaraia cladeNode 323: ch. 5-Snout long. / 1: Snout elongate, frontal, vomer and anterior portion of parasphenoid elongate; preorbital length longer than one-third total head length or greater in mature specimens.ch. 24-Antorbital stripe. / 1: Melanophores absent from narrow band passing lateral to nares.ch. 78-Dorsal margin of frontals. / 1: Portion of frontal anterior to orbit concave in lateral profile.ch. 102-Supratemporal lateralis canal. / 1: Supratemporal laterosensory canal curved at a sharp angle on surface of parietal, extending posterior onto epaxial surface of body; terminal canal pore oriented posteriorly; epidermis overlying supratemporal canal depigmented, forming a pale inverted L shaped patch.ch. 133-Mandibular canal size. / 1: Mandibular canal ossicles dumbbell-shaped.Porotergus+\u201cApteronotus\u201d bonapartii cladeNode 324: ch. 161-Urohyal blade. 0: Posterior blade of urohyal ossified, extending posterior to fourth basibranchial.Apteronotus\u201d bonapartii cladeNode 325: \u201cch. 30-Scales on middorsum. / 1: Scales absent from head, anterior portion of dorsal midline, and area dorsal to pectoral fins.ch. 47-Maxilla descending blade. / 1: Ventral margin of descending blade with a sharp angle about two-thirds distance to its tip; ventral margin posterior to this angle relatively straight.ch. 84-Parasphenoid lateral process. / 0: Lateral margins of parasphenoid extending as broad dorsolateral processes anterior to prootic, extending to a horizontal with trigeminal foramen.ch. 168-Pectoral fin. / 0: Pectoral fin large, more than 43% head length."} +{"text": "Sensors [http://www.mdpi.com/1424-8220/15/5/11769.The authors wish to update the Acknowledgments in their paper published in Sensors , doi:10."} +{"text": "There are errors in the Author Contributions. The correct contributions are: Conceived and designed the experiments: LM MD MES BP ES BS WDH. Performed the experiments: LM MD MES ESC TD KTW. Analyzed the data: LM MD MES ESC BP ES BS TD KTW. Contributed reagents/materials/analysis tools: KTW TD. Wrote the paper: LM WDH. Computational analysis: CL RRR."} +{"text": "From 2001 to 2011, the percentage of children aged <18 years who were receiving special educational or early intervention services increased overall and among Hispanic and non-Hispanic white children, no change was observed among non-Hispanic black children. In 2001 and 2011, Hispanic children were less likely than non-Hispanic white and non-Hispanic black children to receive these services.Sources: Barnes PM, Adams PF, Schiller JS. Summary health statistics for the U.S. population: National Health Interview Survey, 2001. Vital Health Stat 2003;10(217). Available at http://www.cdc.gov/nchs/data/series/sr_10/sr10_217.pdf.http://www.cdc.gov/nchs/data/series/sr_10/sr10_255.pdf.Adams PF, Kirzinger WK, Martinez ME. Summary health statistics for the U.S. population: National Health Interview Survey, 2011. Vital Health Stat 2012;10(255). Available at"} +{"text": "The name of the fourth author was spelled incorrectly. The correct name is: Patrice M. Amb\u00fchl. The correct citation is: Tomonaga Y, Risch L, Szucs TD, Amb\u00fchl PM (2013) The Prevalence of Chronic Kidney Disease in a Primary Care Setting: A Swiss Cross-Sectional Study. PLoS ONE 8(7): e67848. doi:10.1371/journal.pone.0067848."} +{"text": "Nesomyrmex sikorai species-group was assessed via hypothesis-free nest-centroid-clustering combined with recursive partitioning to estimate the number of morphological clusters and determine the most probable boundaries between them. This combination of methods provides a highly automated and objective species delineation protocol based on continuous morphometric data. Delimitations of clusters recognized by these exploratory analyses were tested via confirmatory Linear Discriminant Analysis (LDA) and Multivariate Ratio Analysis (MRA). The final species hypotheses are corroborated by many qualitative characters, and the recognized species exhibit different spatial distributions and occupy different ecological regions. We describe and redescribe eight morphologically distinct species including six new species: Nesomyrmex excelsiorsp. n., N. modestussp. n., N. reticulatussp. n., N. retusispinosus , N. rugosussp. n., N. sikorai , N. striatussp. n., and N. tamatavensissp. n. An identification key for their worker castes using morphometric data is provided.Madagascar is one of the world\u2019s greatest biodiversity hotspots, meriting special attention from biodiversity scientists. It is an excellent testing ground for novel techniques in taxonomy that aim to increase classification objectivity and yield greater taxonomic resolving power. Here we reveal the diversity of a unique and largely unexplored fragment of the Malagasy ant fauna using an advanced combination of exploratory analyses on quantitative morphological data allowing for increased objectivity in taxonomic workflow. The diversity of the The main objective of taxonomy is to document the diversity of flora and fauna. It provides fundamental information for endeavors dealing with biodiversity by addressing a crucial question: \u201chow many species are there?\u201d Unfortunately, the rapidly accelerating rate of biodiversity loss we face today , 2 posesIn recent years a number of promising new algorithmic approaches has been developed and introduced in insect taxonomy for the purpose of recognizing complex patterns in continuous morphometric data \u20137. TheseNesomyrmex sikorai group is inferred via a highly automated protocol involving a fusion of two algorithms, Nest Centroid clustering .CW: Maximum width of the head including compound eyes, [R = 0.999] .CWb: Maximum width of head capsule without the compound eyes. Measured just posterior to the eyes, [R = 0.998] .PoOC: Postocular distance. Use a cross-scaled ocular micrometer and adjust the head to the measuring position of CL. Caudal measuring point: median occipital margin; anterior measuring point: median head at the level of the posterior eye margin, [R = 0.997] .SL: Scape length. Maximum straight line scape length excluding the basal neck and the articular condyle, [R = 0.998] .CS: Absolute cephalic size. The arithmetic mean of CL and CWb.EL: Maximum diameter of the compound eye, [R = 0.929].FRS: Frontal carina distance. Distance of the frontal carinae immediately caudal of the posterior intersection points between frontal carinae and torular lamellae. If these dorsal lamellae do not laterally surpass the frontal carinae, the deepest point of scape corner pits may be taken as the reference line. These pits take up the inner corner of the scape base when the scape is directed fully caudally and produces a dark, triangular shadow in the lateral frontal lobes immediately posterior to the dorsal lamellae of the scape joint capsule, [R = 0.982] .MW: Mesosoma width. In workers MW is defined as the longest width of the pronotum in dorsal view excluding the pronotal spines, [R = 0.998] .PSTI: Apical distance of pronotal spines in dorsal view; if spine tips are rounded or thick take the centers of spine tips as reference points, [R = 0.994] .PEW: Maximum width of petiole in dorsal view. Nodal spines are not considered, [R = 0.996] .PPW: Postpetiole width. Maximum width of postpetiole in dorsal view, [R = 0.994] .SPBA: Minimum propodeal spine distance. The smallest distance of the lateral margins of the propodeal spines at their base. This should be measured in antero-dorsal view, since the wider parts of the ventral propodeum do not interfere with the measurement in this position. If the lateral margins of propodeal spines diverge continuously from the tip to the base, a smallest distance at base is not defined. In this case, SPBA is measured at the level of the bottom of the interspinal meniscus, [R = 0.993] .SPTI: Apical propodeal spine distance. The distance of propodeal spine tips in dorsal view; if spine tips are rounded or truncated, the centers of spine tips are taken as reference points, [R = 0.994] .ML (Weber\u2019s length): Mesosoma length from caudalmost point of propodeal lobe to transition point between anterior pronotal slope and anterior pronotal shield , [R = 0.998] .MPST: Maximum distance from the center of the propodeal spiracle to the posteroventral corner of the ventrolateral margin of the metapleuron, [R = 0.989] .NOH: Maximum height of the petiolar node, measured in lateral view from the uppermost point of the petiolar node perpendicular to a reference line set from the petiolar spiracle to the imaginary midpoint of the transition between the dorso-caudal slope and dorsal profile of caudal cylinder of the petiole, [R = 0.958] .NOL: Length of the petiolar node. Measured in lateral view from the center of the petiolar spiracle to the dorso-caudal corner of caudal cylinder. Do not erroneously take as the reference point the dorso-caudal corner of the helcium, which is sometimes visible, [R = 0.981] .PPL: Postpetiole length. The longest anatomical line that is perpendicular to the posterior margin of the postpetiole and is between the posterior postpetiolar margin and the anterior postpetiolar margin, [R = 0.975] .SPST: Propodeal spine length. Distance between the center of the propodeal spiracle and spine tip. The spiracle center refers to the midpoint defined by the outer cuticular ring but not to the center of the actual spiracle opening, which may be positioned eccentrically, [R = 0.994] .PEH: Maximum petiole height. The longest distance measured from the ventral petiolar profile at node level (perpendicular to the chord length of the petiolar sternum) to the distalmost point of the dorsal profile of the petiolar node, [R = 0.989] .PEL: Diagonal petiolar length in lateral view; measured from anterior corner of subpetiolar process to dorso-caudal corner of caudal cylinder, [R = 0.991] .PPH: Maximum height of the postpetiole in lateral view measured perpendicularly to a line defined by the linear section of the segment border between postpetiolar tergite and sternite, [R = 0.991] .http://purl.org/NET/mx-database). Taxonomic history and descriptions of taxonomic treatments were rendered from this software. Hymenoptera-specific terminology of morphological statements used in descriptions, the identification key, and diagnoses are mapped to classes in phenotype-relevant ontologies (Hymenoptera Anatomy Ontology (HAO) we we12] weNest-centroid clustering (NC-clustering), and linear discriminant analysis (LDA) do not require special data preparation (e.g. standardization), hence raw data were applied for each of the statistical analyses. Data, however, are standardized for the multivariate ratio analysis (MRA) to prevent variables that are larger from dominating the analysis . Variablcluster , petiolar node longer: NOH/CS 0.3 > \u2026 2Scapes very short: SL/CS < 0.73 , petiolar node very short: NOL/CS < 0.3 \u2026 N. reticulatus sp. n.-Propodeal spines triangular, acute, and shorter: SPST/CS < 0.36 \u2026 4Propodeal spines curving downward, blunt, and very long: SPST/CS > 0.36 \u2026 3N. retusispinosus -N. tamatavensis sp. n.Vertex smooth, main sculpture absent, ground sculpture inconspicuously areolate, shiny. In full-face view, compound eyes positioned in the middle of the longitudinal axis of head or closer to the posterior occipital border: PoOC/CL < 0.455 \u2026 Vertex rugose, ground sculpture areolate, dull. In full-face view, compound eyes positioned forward and closer to the anterior clypeal border: PoOC/CL > 0.455 \u2026 -Workers yellow to light brown. Eyes non-protuberant in full-face view: CW/ML: < 0.73 , \u2026 7Black species. Eyes protuberant in full-face view: CW/ML: > 0.73 , \u2026 5N. modestus sp. n.-Main sculpture on vertex and on the dorsum of mesosoma present and coarse areolate or costate, ground sculpture inconspicuously areolate or absent. Eyes strongly protuberant in full-face view: CL/CW < 1.09 \u2026 6Main sculpture on vertex and on the dorsum of mesosoma inconspicuously areolate or absent partly, ground sculpture smooth and shiny. Eyes moderately protuberant in full-face view: CL/CW > 1.09 \u2026 N. sikorai -N. striatus sp. n.Main sculpture on vertex longitudinal costate, ground sculpture inconspicuously areolate. Dorsum of petiolar node smooth and shiny. The best morphometric ratio PSTI/SPTI > 2.5 \u2026 Main sculpture on vertex coarse areolate, ground sculpture inconspicuously areolate. Dorsum of petiolar node rugoso-reticulate, dull. The best morphometric ratio PSTI/SPTI \u2264 2.5 \u2026 N. excelsior sp. n.-N. rugosus sp. n.Postpetiole shorter, anterolateral angles of mesosoma widely distant: PSTI/PPW 1.84 , . Occurs on lower elevations < 500 m. \u2026 Postpetiole longer, anterolateral angles of mesosoma less distant: PSTI/PPW > 1.84 , . Occurs on higher ground > 500 m. \u2026 N. sikorai species-group are described including biogeographic information. The basic statistics of body size ratios are given in In this section, eight species of the urn:lsid:zoobank.org:act:540D306B-7EAC-4AD9-B072-842AC26F91F7Figs .Holotype worker: MADGAGASCAR: Prov. Toliara, For\u00eat Class\u00e9e d'Analavelona, 29.2 km 343\u00b0 NNW Mahaboboka, Madagascar, 1100 m, -22.675 N, 44.19 E, 18.ii.2003, collection code: BLF07826; CASENT0498265, Fisher et al., ex dead twig above ground montane rainforest ;Paratypes: 8 workers and a single gyne with the same label data with the holotype under CASENT codes: CASENT0763558, BLF07826, ; CASENT0498263, BLF07826, ; CASENT0498264, BLF07826, ; CASENT0498266, BLF07826, ;The list of 83 worker individuals belonging to 58 nest samples morphometrically investigated is given in See in key.Body color: yellow; brown. Body color pattern: Body concolorous. Absolute cephalic size: 726 \u03bcm . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.281 . Postocular distance vs. cephalic length (PoOc/CL): 0.457 . Postocular sides of cranium contour, anterior view orientation: converging posteriorly. Postocular sides of cranium contour, anterior view shape: convex. Vertex contour line in anterior view shape: straight; feebly convex. Vertex sculpture: main sculpture rugoso-reticulate, ground sculpture smooth. Gena contour line in anterior view shape: convex. Genae contour from anterior view orientation: strongly converging. Gena sculpture: rugoso-reticulate with feeble areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.233 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.289 . Longitudinal carinae on median region of frons count: present. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.822 . Median clypeal notch count: present. Median carina of clypeus count: absent. Spine length vs. absolute cephalic size (SPST/CS): 0.275 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.225 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.252 . Propodeal spine shape: straight; triangular, blunt. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.607 . Metanotal depression count: present. Dorsal region of mesosoma sculpture: rugose with smooth ground sculpture. Lateral region of pronotum sculpture: inconspicuously areolate ground sculpture, main sculpture rugoso-reticulate. Mesopleuron sculpture: smooth ground sculpture, superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.233 . Dorsal region of petiole sculpture: ground sculpture inconspicuously areolate, main sculpture rugoso-reticulate. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.354 . Dorsal region of postpetiole sculpture: ground sculpture smooth, main sculpture dispersed rugose.The name refers to the typically montane occurrence of this species.This species is known to occur in smaller, isolated montane rainforests between elevation of 520 m and 1325 m (mean: 1007 m) in the western and central part of Madagascar .urn:lsid:zoobank.org:act:A8B6069A-2FD3-4CEA-AC0D-DB36127037B0Figs .Holotype worker: MADGAGASCAR: Prov. Toliara, Anosy Region, Anosyenne Mts, 29.33 km NW Manantenina, Madagascar, 540 m, -24.13993 N, 47.07418 E, 21.ii.2015, collection code: BLF36218; CASENT0393175, B.L.Fisher, F.A.Esteves et al. ;Paratypes: 13 workers with the same label data with the holotype under CASENT codes: CASENT0393167, BLF36224, ; CASENT0393168, BLF36224, ; CASENT0393169, BLF36224, ; CASENT0393170, BLF36224, ; CASENT0393171, BLF36224, ; CASENT0393172, BLF36224, ; CASENT0393173, BLF36224, ; CASENT0393176, BLF36218, ; CASENT0393177, BLF36218, ; CASENT0393178, BLF36218, ; CASENT0393179, BLF36218, ; CASENT0393180, BLF36218, ; CASENT0393181, BLF36218, ;The list of 18 worker individuals belonging to 18 nest samples morphometrically investigated is given in See in key.Body color: black. Body color pattern: concolorous. Absolute cephalic size: 796 \u03bcm . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.216 . Postocular distance vs. cephalic length (PoOc/CL): 0.453 . Postocular sides of cranium contour, anterior view orientation: converging posteriorly. Postocular sides of cranium contour, anterior view shape: convex. Vertex contour line in anterior view shape: straight; feebly convex. Vertex sculpture: main sculpture areolate, ground sculpture smooth. Gena contour line in anterior view shape: convex. Gena contour from anterior view orientation: strongly converging. Gena sculpture: rugoso-reticulate with feeble areolate ground sculpture; rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: present. Eye length vs. absolute cephalic size (EL/CS): 0.234 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.295 . Longitudinal carinae on median region of frons count: present. Smooth median region on frons count: present. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.771 . Median clypeal notch: present. Median carina of clypeus count: absent. Spine length vs. absolute cephalic size (SPST/CS): 0.236 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.236 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.251 . Propodeal spine shape: triangular, blunt. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.657 . Metanotal depression count: present. Dorsal region of mesosoma sculpture: smooth ground sculpture superimposed by feeble areolate main sculpture. Lateral region of pronotum sculpture: inconspicuous areolate ground sculpture, main sculpture dispersed costate. Mesopleuron sculpture: smooth ground sculpture, superimposed by dispersed rugulae. Metapleuron sculpture: fine areolate ground sculpture, superimposed by dispersed rugulae. Petiole width vs. absolute cephalic size (PEW/CS): 0.223 . Dorsal region of petiole sculpture: ground sculpture smooth, main sculpture absent. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.341 . Dorsal region of postpetiole sculpture: ground sculpture smooth, main sculpture absent.The name (modestus = moderate) refers to the moderately coarse surface sculpturing of this species.This species is known to occur in rainforests between elevation of 520 m and 1325 m (mean: 514 m) in the southwestern part of Madagascar .urn:lsid:zoobank.org:act:3E5E26A6-6795-45AA-A051-2AFDAF57E6E5Figs .Holotype worker: MADGAGASCAR: Prov. Toliara, For\u00eat de Kirindy, 15.5 km 64\u00b0 ENE Marofandilia, 100 m, -20.045 N, 44.66222 E, 28.xi.2001, collection code: BLF04604; CASENT0418497, Fisher-Griswold Arthropod Team, beating low vegetation tropical dry forest ;Paratypes: 3 workers and a gyne with the same label data with the holotype under CASENT codes: CASENT0473935, BLF04605, ; CASENT0418443, BLF04604, ; CASENT0418447, BLF04604, ; CASENT0418482, BLF04604, ;The list of 7 worker individuals belonging to 7 nest samples morphometrically investigated is given in See in key.Body color: yellow; brown. Body color pattern: concolorous. Absolute cephalic size: 511 \u03bcm . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.151 . Postocular distance vs. cephalic length (PoOc/CL): 0.462 . Postocular sides of cranium contour, anterior view orientation: converging posteriorly. Postocular sides of cranium contour, anterior view shape: strongly convex. Vertex contour line in anterior view shape: slightly concave. Vertex sculpture: main sculpture homogeneously forked costate, ground sculpture areolate. Gena contour line in anterior view shape: convex. Gena contour from anterior view orientation: strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: present. Eye length vs. absolute cephalic size (EL/CS): 0.238 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.322 . Longitudinal carinae on median region of frons: absent. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.704 . Median clypeal notch: present. Median carina of clypeus: absent. Spine length vs. absolute cephalic size (SPST/CS): 0.236 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.254 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.267 . Propodeal spine shape: straight; triangular, blunt. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.629 . Metanotal depression: present. Dorsal region of mesosoma sculpture: areolate ground sculpture, superimposed by dispersed rugae. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture dispersed costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Petiole width vs. absolute cephalic size (PEW/CS): 0.235 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture absent. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.37 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture absent.The name refers to the fine, micro-reticulate body sculpturing.This species is known to occur in dry forests in lowlands between 50 m and 130 m (mean: 94 m) of the western and southern coasts of Madagascar .Figs .Holotype worker: \u201cL. retusispinosus, type, Forel, For d' andrangoloaca , Madagascar (Sikora)\u201d, ;The list of 9 worker individuals belonging to 9 nest samples morphometrically investigated is given in See in key.Body color: yellow; brown. Body color pattern: concolorous. Absolute cephalic size: 812 \u03bcm . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.17 . Postocular distance vs. cephalic length (PoOc/CL): 0.483 . Postocular sides of cranium contour, anterior view orientation: converging posteriorly. Postocular sides of cranium contour, anterior view shape: convex. Vertex contour line in anterior view shape: straight; feebly convex. Vertex sculpture: main sculpture rugoso-reticulate, ground sculpture areolate. Gena contour line in anterior view shape: convex. Gena contour from anterior view orientation: strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: present. Eye length vs. absolute cephalic size (EL/CS): 0.237 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.294 . Longitudinal carinae on median region of frons: present. Smooth median region on frons: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.838 . Median clypeal notch: present. Median carina of clypeus: present or absent. Spine length vs. absolute cephalic size (SPST/CS): 0.43 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.24 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.322 . Propodeal spine shape: slightly or strongly bent. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.659 . Metanotal depression: present. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture rugoso-reticulate. Mesopleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Metapleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.24 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.39 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.This species is known to occur in rain forests and montane forests in lowlands between 918 m and 1080 m in central Madagascar .urn:lsid:zoobank.org:act:C39EB560-EC60-4720-B6D8-95CE88C75A4DFigs .Holotype worker: MADGAGASCAR: Prov. Antsisarana, Masoala National Park, 250 m, -15.33058 N, 50.30279 E, 13.iii.2014, collection code: BLF33157; CASENT0374465, Fisher et al. ;Paratypes: 2 workers, a gyne and a male with the same label data with the holotype under CASENT codes: CASENT763560, BLF33157, , CASENT0374466, BLF33157, ; CASENT0377059, BLF32874, ;The list of 24 worker individuals belonging to 23 nest samples morphometrically investigated is given in See in key.Body color: yellow to brown. Body color pattern: concolorous. Absolute cephalic size: 747 \u03bcm . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.239 . Postocular distance vs. cephalic length (PoOc/CL): 0.467 . Postocular sides of cranium contour, anterior view orientation: converging posteriorly. Postocular sides of cranium contour, anterior view shape: convex. Vertex contour line in anterior view shape: straight or feebly convex. Vertex sculpture: main sculpture rugoso-reticulate, ground sculpture smooth. Gena contour line in anterior view shape: convex. Genae contour from anterior view orientation: strongly converging. Gena sculpture: rugoso-reticulate with feeble areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: present. Eye length vs. absolute cephalic size (EL/CS): 0.226 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.292 . Longitudinal carinae on median region of frons: present. Smooth median region on frons: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.816 . Median clypeal notch count: present. Median carina of clypeus: absent. Spine length vs. absolute cephalic size (SPST/CS): 0.288 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.221 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.257 . Propodeal spine shape: straight. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.635 . Metanotal depression: present. Dorsal region of mesosoma sculpture: rugose with smooth ground sculpture. Lateral region of pronotum sculpture: inconspicuously areolate ground sculpture, main sculpture rugoso-reticulate. Mesopleuron sculpture: smooth ground sculpture, superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.231 . Dorsal region of petiole sculpture: ground sculpture inconspicuously areolate, main sculpture rogoso-reticulate. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.345 . Dorsal region of postpetiole sculpture: ground sculpture smooth, main sculpture dispersed rugose.The name refers to the coarse, rugose body sculpturing.This species is known to occur in rainforests between elevation of 200 m and 520 m (mean: 390 m) of the eastern and southern coasts of Madagascar .Figs .Holotype worker: \u201cLeptothorax sikorai n. sp., Imerina , Sikora\u201d TYPUS . Note: the type was not available for morphometric investigation; high-quality AntWeb images were used for comparison.The list of 36 worker individuals belonging to 28 nest samples morphometrically investigated is given in See in key.Body color: black. Body color pattern: concolorous. Absolute cephalic size: 822 \u03bcm . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.169 . Postocular distance vs. cephalic length (PoOc/CL): 0.488 . Postocular sides of cranium contour, anterior view orientation: converging posteriorly. Postocular sides of cranium contour, anterior view shape: strongly convex. Vertex contour line in anterior view shape: straight to feebly convex. Vertex sculpture: main sculpture areolate, ground sculpture areolate. Gena contour line in anterior view shape: convex. Gena contour from anterior view orientation: strongly converging. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: present. Eye length vs. absolute cephalic size (EL/CS): 0.212 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.292 . Longitudinal carinae on median region of frons: present. Smooth median region on frons: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.828 . Median clypeal notch: present. Median carina of clypeus: present or absent. Spine length vs. absolute cephalic size (SPST/CS): 0.269 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.253 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.299 . Propodeal spine shape: straight; triangular, blunt. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.665 . Metanotal depression: present. Dorsal region of mesosoma sculpture: areolate main sculpture, interstices areolate. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture areolate. Mesopleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Metapleuron sculpture: areolate ground sculpture, superimposed by coarse rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.269 . Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture rugoso-reticulate. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.37 . Dorsal region of postpetiole sculpture: ground sculpture smooth, main sculpture absent.This species is known to occur in montane rainforests between elevations of 200 m and 520 m in central Madagascar .urn:lsid:zoobank.org:act:538BA1CB-262A-492E-86E5-035513668BE9Figs .Holotype worker: MADGAGASCAR: Prov. Toliara, Anosy Region, Anosyenne Mts., 31.2 km NW Manantenina, 1125 m, -24.13894 N, 47.06804 E, 26.ii.2015, collection code: BLF36533; CASENT0393121, Fisher et al. ;Paratypes: 5 workers and a male with the same label data with the holotype under CASENT codes: CASENT0393116, BLF36533, ; CASENT0393117, BLF36533, ; CASENT0393118, BLF36533, ; CASENT0393119, BLF36533, ; CASENT0393120, BLF36533, ;The list of 9 worker individuals belonging to 9 nest samples morphometrically investigated is given in See in key.Body color: black. Body color pattern: concolorous. Absolute cephalic size: 842 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.155 . Postocular distance vs. cephalic length (PoOc/CL): 0.460 . Postocular sides of cranium contour, anterior view orientation: converging posteriorly. Postocular sides of cranium contour, anterior view shape: convex. Vertex contour line in anterior view shape: straight to feebly convex. Vertex sculpture: main sculpture parallel costate, ground sculpture smooth. Gena contour line in anterior view shape: convex. Gena contour from anterior view orientation: strongly converging. Gena sculpture: rugoso-reticulate with feeble areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: present. Eye length vs. absolute cephalic size (EL/CS): 0.219 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.288 . Longitudinal carinae on median region of frons: present. Smooth median region on frons: present. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.794 . Median clypeal notch count: present. Median carina of clypeus: absent. Spine length vs. absolute cephalic size (SPST/CS): 0.241 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.242 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.244 . Propodeal spine shape: curving upward. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.666 . Metanotal depression: present. Dorsal region of mesosoma sculpture: costate with smooth ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture areolate. Mesopleuron sculpture: smooth ground sculpture, superimposed by dispersed rugulae. Metapleuron sculpture: fine areolate ground sculpture, superimposed by coarse rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.243 . Dorsal region of petiole sculpture: ground sculpture smooth, main sculpture absent. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.362 . Dorsal region of postpetiole sculpture: ground sculpture smooth, main sculpture absent.The name refers to the coarse, striate/costate surface sculpturing on the head and mesosoma.This species is known to occur in montane rainforests between elevations of 900 m and 1125 m (mean: 1054 m) of the southeastern part of Madagascar .urn:lsid:zoobank.org:act:A67F58B6-502A-4798-B638-6FB35C78BD27Figs .Holotype worker: MADGAGASCAR: Prov. Toamasina, Montagne d'Akirindro 7.6 km 341\u00b0 NNW Ambinanitelo, 600 m, -15.28833 N, 49.54833 E, 17.iii.2003, collection code: BLF8390; CASENT0496292, Fisher et al., ex dead twig above ground, rainforest ;Paratypes: 5 workers and a male with the same label data with the holotype with CASENT codes: CASENT0763559, BLF8390, ; CASENT0496293, BLF8390, ; CASENT0496294, BLF8390, ; CASENT0496295, BLF8390, ;The list of 41 worker individuals belonging to 32 nest samples morphometrically investigated is given in See in key.Body color: yellow to brown. Body color pattern: concolorous. Absolute cephalic size: 699 \u03bcm . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.155 . Postocular distance vs. cephalic length (PoOc/CL): 0.433 . Postocular sides of cranium contour, anterior view orientation: converging posteriorly. Postocular sides of cranium contour, anterior view shape: convex. Vertex contour line in anterior view shape: straight to feebly convex. Vertex sculpture: main sculpture inconspicuous, ground sculpture smooth. Gena contour line in anterior view shape: convex. Gena contour from anterior view orientation: strongly converging. Gena sculpture: rugoso-reticulate with feeble areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen: present. Eye length vs. absolute cephalic size (EL/CS): 0.258 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.329 . Longitudinal carinae on median region of frons: present or absent. Smooth median region on frons: present. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.798 . Median clypeal notch: present. Median carina of clypeus: absent. Spine length vs. absolute cephalic size (SPST/CS): 0.388 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.244 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.303 . Propodeal spine shape: strongly bent. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.739 . Metanotal depression: present. Dorsal region of mesosoma sculpture: rugose with smooth ground sculpture. Lateral region of pronotum sculpture: inconspicuously areolate ground sculpture, main sculpture rugoso-reticulate. Mesopleuron sculpture: smooth ground sculpture, superimposed by dispersed rugulae. Metapleuron sculpture: fine areolate ground sculpture, superimposed by dispersed rugulae. Petiole width vs. absolute cephalic size (PEW/CS): 0.243 . Dorsal region of petiole sculpture: ground sculpture inconspicuously areolate, main sculpture rugoso-reticulate. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.363 . Dorsal region of postpetiole sculpture: ground sculpture smooth, main sculpture dispersed rugose.Tamatave is the French name of Toamasina, the capital of the Atsinanana region, where this species is abundant.This species is known to occur in rainforests from sea level to 1100 m predominantly in the northeastern part of Madagascar . The onlS1 Appendix(NEX)Click here for additional data file.S2 AppendixMark dendrogram function mapping the results of partitioning algorithm PART on the dendrogram is also added.(TXT)Click here for additional data file.S1 TableUnique CASENT number for pinned samples, locality, geographic coordinates in decimal format, altitude (ALT) in meters a.s.l., collector\u2019s name, date and number of specimens investigated bearing the given CASENT number are provided. HT = Holotype, PT = paratype(s). All samples collected in Toliara administrative region, Madagascar, and deposited at the California Academy of Sciences (CAS).(XLSX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 TableCASENT code (casent), final species hypothesis (species), geographic coordinates and the name format as samples appear on the dendrogram (dendro-name) are also provided in the table. HT = Holotype, PT = paratype(s).(CSV)Click here for additional data file."} +{"text": "Caenorhabditis elegans Fast Bioassay. PLoS ONE 10(6): e0128898. doi:10.1371/journal.pone.0128898. The publisher apologizes for this error.The third author\u2019s name is spelled incorrectly. The correct name is: Lucila Ines Buzzi. The correct citation is: Bianchi JI, Stockert JC, Buzzi LI, Bl\u00e1zquez-Castro A, Simonetta SH (2015) Reliable Screening of Dye Phototoxicity by Using a"} +{"text": "Aging (Albany NY) 2015; 8(3): 521-538.PMCID: PMC 833143 PMID: 26946062http://www.impactaging.com/papers/v8/n3/pdf/100913.pdf. The correct corresponding authors are provided here:In this Article, the information on corresponding author is wrong: wdzhangy@hotmail.com; shanleicn@126.comCorrespondence: Weidong Zhang, PhD; Lei Shan, PhD; E-mail:"} +{"text": "Service, imaging, open source, archive, image sharing, clinical trials.http://www.xnat.org) ; documentation of RSNA CTP [http://mircwiki.rsna.org/index.php?title=CTP_Articles].Documentation and links to both XNAT instances and Puppet installation scripts ["} +{"text": "Drosophila\u00a0Microbiota Modulates Host Metabolic Gene Expression via IMD/NF-\u03baB Signaling. PLoS ONE 9(4): e94729. doi:10.1371/journal.pone.0094729The abbreviation of the first author\u2019s name is incorrect in the citation. The correct abbreviation is: Erkosar B. The correct citation is: Erkosar B, Defaye A, Bozonnet N, Puthier D, Royet J, et al. (2014)"} +{"text": "Eucalyptus Subgenus Eucalyptus (Myrtaceae) Are a Rich Source of Flavonoids and Related Non-Volatile Constituents. PLoS ONE 11(3): e0151432. doi:10.1371/journal.pone.0151432The second author\u2019s last name is spelled incorrectly. The correct name is: Samiddhi L. Senaratne. The correct citation is: Goodger JQD, Senaratne SL, Nicolle D, Woodrow IE (2016) Foliar Essential Oil Glands of"} +{"text": "Nature Communications7: Article number: 11311 10.1038/ncomms11311 (2016); Published: 04182016; Updated: 06032016The author Yueh-Hsiung Kuo was incorrectly omitted from the list of corresponding authors in this Article. The correct information for correspondence is: \u2018Correspondence and requests for materials should be addressed to Y.-H.K. kuoyh@mail.cmu.edu.tw) or to N.S.Y. nsyang@gate.sinica.edu.tw)'."} +{"text": "Nature Communications7: Article number: 1211210.1038/ncomms12112 (2016); Published: 06292016; Updated: 08032016.The present address for Ioana C. G\u00e2rlea is incorrect in this Article. The correct present address for this author is given below:Faculty of Physics, University of Vienna, Boltzmanngasse 5, A-1090 Vienna, Austria.The name of this author is also incorrect in the \u2018How to cite this article' section. This section should read:et al. Finite particle size drives defect-mediated domain structures in strongly confined colloidal liquid crystals. Nat. Commun. 7:12112 doi: 10.1038/ncomms12112 (2016).G\u00e2rlea, I. C."} +{"text": "Determining the evolutionary relationships among the major lineages of extant birds has been one of the biggest challenges in systematic biology. To address this challenge, we assembled or collected the genomes of 48 avian species spanning most orders of birds, including all Neognathae and two of the five Palaeognathae orders. We used these genomes to construct a genome-scale avian phylogenetic tree and perform comparative genomic analyses.Here we present the datasets associated with the phylogenomic analyses, which include sequence alignment files consisting of nucleotides, amino acids, indels, and transposable elements, as well as tree files containing gene trees and species trees. Inferring an accurate phylogeny required generating: 1) A well annotated data set across species based on genome synteny; 2) Alignments with unaligned or incorrectly overaligned sequences filtered out; and 3) Diverse data sets, including genes and their inferred trees, indels, and transposable elements. Our total evidence nucleotide tree (TENT) data set gave what we consider our most reliable species tree when using the concatenation-based ExaML algorithm or when using statistical binning with the coalescence-based MP-EST algorithm (which we refer to as MP-EST*). Other data sets, such as the coding sequence of some exons, revealed other properties of genome evolution, namely convergence.The Avian Phylogenomics Project is the largest vertebrate phylogenomics project to date that we are aware of. The sequence, alignment, and tree data are expected to accelerate analyses in phylogenomics and other related areas.The online version of this article (doi:10.1186/s13742-014-0038-1) contains supplementary material, which is available to authorized users. Here we present FASTA files of loci, sequence alignments, indels, transposable elements, and Newick files of gene trees and species trees used in the Avian Phylogenomics Project -4. We alHere we describe each locus data set in brief. Additional details are provided in Jarvis et al. .This is an exon-coding sequence data set of 8295 genes based on synteny-defined orthologs we identified and selected from the assembled genomes of chicken and zebra finch [These are alignments of the translated peptide sequences for the 8295 protein-coding gene data set.This is an orthologous subset of introns from the 8295 protein-coding genes among 52 species (includes outgroups). Introns with conserved annotated exon-intron boundaries between chicken and another species (\u00b11 codon) were chosen. We filtered out introns with length\u2009<\u200950\u00a0bp or intron length ratio\u2009>\u20091.5 between chicken and another species or another species and chicken. This filtering resulted in a conservative subset of introns that could be reliably identified and aligned.This is the ultraconserved element (UCE) data set with 1000\u00a0bp flanking sequence at the 3\u2032 and 5\u2032 ends. The UCE dataset was filtered to remove overlap with the above exon and intron data sets, other exons and introns in the chicken genome assembly version 3, and overlapping sequences among the UCEs. The source UCE sequences used to search the genomes were determined from sequence capture probes aligned These four data sets represent the 10% subsets of the 8295 exons and their associated introns when available (i.e. from the same genes) that had the highest and lowest variance in GC3 (third codon position) content across species. To calculate GC3 variance, we first calculated GC3 for each ortholog in each species, and then we used the correlation coefficient R to calculate variance in GC3 for each species. Orthologs were ranked by their GC3 variance and we selected the top and bottom 10% for analyses.These are the concatenated sets of loci from various partitions of the TENT dataset , brought together using the statistical binning approach. The statistical binning approach put together sets of loci that were deemed \u201ccombinable\u201d. Two genes were considered combinable if their respective gene trees had no pairs of incompatible branches that had bootstrap support above a 50% threshold. Alignments of genes in the same bin were concatenated to form supergenes, but boundaries of genes were kept so that a gene-partitioned phylogenetic analysis could be performed on each supergene.http://www.bx.psu.edu/miller_lab/) across all 48 bird species and outgroups using individual chromosomes of the chicken genome as the reference . They were filtered for segments with fewer than 42 avian species (>5 missing bird species) and aberrant sequence alignments. The individual remaining segments of the MULTIZ alignment were realigned with MAFFT. We did not use SAT\u00e9\u2009+\u2009MAFFT due to computational challenges (too much input/output was required).Whole genome alignments were first created by a LASTZ\u2009+\u2009MULTIZ alignment ,14 (http5.7 million insertions and deletions (indels) were scored as binary characters locus by locus from the same intron, exon, and UCE alignments as used in the TENT data set on the principle of simple indel coding using 2Xread ,16 and tThese are 61 manually curated presence/absence loci of transposable elements (TEs) present in the Barn Owl genome that exhibit presence at orthologous positions in one or more of the other avian species. The TE markers were identified by eye after a computational screening of 3,671 TguLTR5d retroposon insertions from the Barn Owl. For each TguLTR5d locus, we conducted BLASTn searches of TE-flanking sequences (1\u00a0kb per flank) against the remaining avian species and generated multispecies sequence alignments using MAFFT . RedundaWe provide the above loci data sets as FASTA files of both unfiltered and filtered sequence alignments. The alignments were filtered for aberrant over- and under-aligned sequences, and for the presence of the loci in 42 of the 48 avian species. All multiple sequence alignments were performed in two rounds. The first round was used to find contiguous portions of sequences that we identified as aberrant, and the second round was used to realign the filtered sequences. We used SAT\u00e9 ,23 combiThese are filtered alignments of exons from 8295 genes. Of these 8295, there were 42 genes that were identified to have annotation issues and we removed them from the phylogenetic analyses . Two more genes were removed because a gene tree could not be estimated for them. The first round of alignment was performed using SAT\u00e9\u2009+\u2009PRANK, and the second round was performed using SAT\u00e9\u2009+\u2009MAFFT. Before alignment, the nucleotide sequences were converted to amino acid sequences, and then reverted back to nucleotide sequences afterwards.42-exon-genes-removed.txt: list of 42 genes removed due to various issuespep2cds-filtered-sate-alignments-noout.tar.gz: DNA alignments without outgroupspep2cds-filtered-sate-alignments-original.zip: DNA alignments with outgroups included8295 Exonspep-filtered-sate-alignments-noout.tar.gz: Amino acid alignments with outgroups removedpep-filtered-sate-alignments-original.zip: Amino acid alignments with outgroups included8295 Amino AcidsThese are filtered alignments of introns from 2516 genes. Both rounds of alignment were performed using SAT\u00e9\u2009+\u2009MAFFT, because SAT\u00e9\u2009+\u2009PRANK was too computationally expensive on long introns.introns-filtered-sate-alignments-with-and-without-outgroups.tar.gz: Includes both alignments with and without outgroups2516 IntronsThese are alignments of UCEs and their surrounding 1000\u00a0bp from 3769 loci after filtering. Both rounds of alignment were performed using SAT\u00e9\u2009+\u2009MAFFT.uce-probes-used.fasta.gz: Probes targeting UCE loci shared among vertebrate taxa.uce-raw-genome-slices-of-probe-matches.tar: Probe\u2009+\u2009flank slices around locations matching probes targeting UCE loci.uce-raw-lastz-results-of-probe-matches.tar: LASTZ results of mapping probes onto genome assemblies.uce-assembled-loci-from-probe-matches.tar: UCE loci assembled from probe\u2009+\u2009flank slices from each genome.uce-filtered-alignments-w-gator.tar.gz: UCE individual alignments without outgroupsuce-filtered-alignments-without-gator.tar.gz: UCE individual alignments with outgroups3769 UCE\u2009+\u20091000 flanking bpsupergene-alignments.tar.bz2: supergene alignments with partition files showing genes put in each bin and their boundaries in the concatenated alignmentThese are concatenated alignments for each of our 2022 supergene alignments. We note that although supergenes are concatenated loci, we estimated supergene trees using partitioned analyses where each gene was put in a different partition. Thus, we also provide the boundaries between genes in text files (these can be directly used as partition input files to RAxML).These are individual loci alignments of the above data sets, before filtering.pep-unfiltered-alignments-original.zip: unfiltered SAT\u00e9\u2009+\u2009Prank alignments used for the filtering stepAmino.Acid.unfilteredpep2cds-unfiltered-alignemtns-original.zip: unfiltered SAT\u00e9\u2009+\u2009Prank alignments used for the filtering stepExon.c123.unfiltered:introns-unfiltered-alignments-original.zip: intron SAT\u00e9 alignments before filtering with outgroups includedintrons-unfiltered-alignments-noout.zip: intron SAT\u00e9 alignments before filtering with outgroups includedIntron.unfiltereduce-unfiltered-alignments-w-gator.tar.gz: UCE alignments before filtering with alligator outgroupUCE.unfilteredhttps://github.com/gigascience/paper-zhang2014.These are uploaded as part of the comparative genomics paper data notWGT.unfilteredWe provide FASTA files of concatenated sequence alignments of the above filtered loci datasets. These are concatenated alignments that were used in the ExaML and RAxML analyses .Exon.AminoAcid.ExaML.partitionedExon.c123. ExaML.partitionedExon.c123. ExaML.unpartitionedExon.c1.ExaML.unpartitionedExon.c2.ExaML.unpartitionedExon.c12.ExaML.unpartitionedExon.c123-RY.ExaML.unpartitionedExon.c3.ExaML.unpartitionedIntronTEIT.RAxMLTENT\u2009+\u2009c3.ExaMLTENT\u2009+\u2009outgroup.ExaMLTENT.ExaML.100%TENT.ExaML.25%TENT.ExaML.50%TENT.ExaML.75%WGT.ExaMLuce-filtered-alignments-w-gator-concatenated.phylip.gzuce-filtered-alignments-without-gator-concatenated.phylip.gzUCE concatenated alignments with and without the alligatorc12.DNA.alignment.1156.clocklike.zipc12.DNA.alignment.1156.clocklike.txtc12.DNA.alignment.clocklike.readme.txtc12.DNA.alignment.clocklike.txt.zipConcatenated c12 (1st\u2009+\u20092nd codons) DNA sequence alignments from the 1156 clocklike genes were used for the dating analyses. These are alignments of the first and second codon positions of clock-like genes among the 8295 exon orthologs:High variance exons:Exon.heterogeneous.c123Exon.heterogenous.c12Low variance exons:Exon.homogeneous.c123.Exon.homogenous.c12High variance introns: These are heterogenous intronsconcatIntronNooutMSAlow.fasta.gzLow variance introns: These are homogenous intronsconcatIntronNooutMSAhigh.fasta.gzThis is a concatenated alignment of indels from exons, introns, and UCEs. A README file describes the content.owl_TE_marker_Table.txtSpecies trees (Newick format) were generated with either RAxML, an improved ExaML version for handling large alignments, or MP-EST* . We depoExon.AminoAcid.ExaML.partitioned.treExon.c123.ExaML.partitioned.treExon.c123.ExaML.unpartititoned.treExon.c123-RY.ExaML.unpartitioned.treExon.c12.ExaML.partitioned.treExon.c12.ExaML.unpartitioned.treExon.c1.ExaML.unpartitioned.treExon.c2.ExaML.unpartitioned.treExon.c3.ExaML.unpartitioned.treExon.RAxML.heterogenous.c123.treExon.RAxML.heterogenous.c12.treExon.RAxML.homogenous.c123.treExon.RAxML.homogenous.c12.treIntron.RAxML.heterogenous.tre.txtIntron.RAxML.homogenous.tre.txtIntron.RAxML.partitioned.treIntron.RAxML.unpartitioned.treIntron.MP-EST.binned.treIntron.MP-EST.unbinned.treTEIT.RAxML.treTENT\u2009+\u2009c3.ExaML.treTENT\u2009+\u2009outgroup.ExaML.treTENT.ExaML.100%.treTENT.ExaML.25%.treTENT.ExaML.50%.treTENT.ExaML.75%.treUCE.RAxML.unpartitioned.treWGT.ExaML.alternative.treWGT.ExaML.best.treeChronogram01.TENT.ExAML.treChronogram02.TENT.ExAML.max865.treChronogram03.TENT.ExAML.Allig247.treChronogram04.TENT.ExAML.no-outgroup.treChronogram05.TENT.ExAML.no-outgroup.max865.treChronogram06.TENT.MP-EST.treChronogram07.WGT.ExAML.alternative.treChronogram08.WGT.ExAML.best.treChronogram09.Intron.ExAML.unpartitioned.treChronogram10.UCE.RAxML.treChronogram11.Exon.c123.RaXML.partitioned.treML (bestML) gene treesBootstrap replicates of ML gene treesML (bestML) supergene trees used in MP-EST analysesBootstrap replicates of supergene trees used in MP-EST analysesPartition files showing which loci make up which bins for MP-EST analysesScript for filtering amino acid alignmentsScript for filtering nucleotide sequence alignmentsScript for mapping names from 5-letter codes to full namesScripts related to indel analysesWe also deposit the key scripts used in this project in GigaDB, which include:We provide readme files in the script directories describing the usage of the scripts.Project name: Avian Phylogenomic Project scriptshttps://github.com/gigascience/paper-jarvis2014; also see companion paper home page for related data https://github.com/gigascience/paper-zhang2014Project home page: Operating system: UnixProgramming language: R, Perl, pythonLicense: GNU GPL v3.Any restrictions to use by non-academics: noneGigaScience repository, GigaDB [http://avianbase.narf.ac.uk/index.html UCSC: CoGe: (https://genomevolution.org/wiki/index.php/Bird_CoGe).Other data files presented in this data note for the majority of genomes are available in the , GigaDB (Table\u00a01"} +{"text": "Temnothorax nylanderi species-group of myrmicine ants is characterized. Eighteen species belonging to this group in the Ponto-Mediterranean region are described or redefined based on an integrative approach that combines exploratory analyses of morphometric data and of a 658bp fragment of the mitochondrial gene for the cytochrome c oxidase subunit I (CO I). The species group is subdivided into five species complexes: T. angustifrons complex, T. lichtensteini complex, T. nylanderi complex, T. parvulus complex, T. sordidulus complex, and two species, T. angulinodissp. n. and T. flavicornis form their own lineages. We describe seven new species , raise T. tergestinus stat.n. to species level, and propose a new junior synonymy for T. saxonicus syn.n. (junior synonym of T. tergestinus). We describe the worker caste and provide high quality images and distributional maps for all eighteen species. Furthermore, we provide a decision tree as an alternative identification key that visually gives an overview of this species-group. We make the first application to Formicidae of the Semantic Phenotype approach that has been used in previous taxonomic revisions.In the current revisionary work, the Temnothorax (formerly synonymized with Leptothorax) is one of the most speciose ant genera on earth, with 380 valid species and 47 subspecies worldwide (http://www.antcat.org). It is especially well represented in the European fauna. Members of the genus occur in a great variety of habitats and display a considerable range of social behavior , ne, neTemnon making , 10], d, dTemnotn making , the regn making , 13], , , , , , , 25] th th25] thic areas , 27], . T. TTemnothodology , 31], . I. ITemnoter group , 35]..TemnothoT. nylanderi species-group in the Ponto-Mediterranean region is coupled with a large number of cryptic species , w, wT. nyl species , using mIn our study we initially recognized morphological patterns using the exploratory data analysis tool NC-clustering , which aTemnothorax and help to better understand the biogeographic patterns of this genus in the Ponto-Mediterranean region.Our data reveal cryptic diversity in this species-group of the genus type material investigated. Non-type material that was morphometrically examined in this revision is listed in GRE:Levidi-10S-20000427-123) generated from the original label information as follows:In the present study, we recorded 22 continuous morphometric traits in 1693 worker individuals belonging to 526 nest samples. Specimens for the present study were borrowed from public and private collections, see the list of institutions below. Our study did not involve endangered or protected species. Label information of type material is given for each taxon in the section A three-letter country code , the nearest settlement given in the label (separated from the following parts by hyphens), the distance and direction from it, the sampling date in alpha-numeric format and a final unique field sample identifier.http://www.antweb.org) and can be uniquely identified by their specimen-level codes affixed to each pin. Information about the taxonomic history of the taxa redefined below is based on B. Bolton in AntCat (http://www.antcat.org). Distribution maps were generated by QGIS 2.4.0 software http://prHereby we propose two new template EQ expression as an expansion of the semantic statement types described by Balhoff et al. and Mik\u00f3Angle between anatomical linesAngles between anatomical lines are often used in taxonomic treatments of Formicidae:NL: Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 72\u201382\u00b0\u201dWe expressed angle value between anatomical lines using PATO\u2019s angle and UO\u2019s degree with the combination of our previously proposed relative and absolute measurement phenotypes:SP: \u2018has part' some and ))) and (towards some 'anterior profile of petiolar node contour line'))))ColorationWe present color hue and intensity as separated entity attributes:NL: Body color: brownSP: 'has part' some (body and ('bearer of' some brown))andNL: Body color pattern: mesosoma, antenna and legs excluding femora, waist and anterior region of 1st gastral tergite lighter than head, femora and posterior region of gaster.SP: ('has part' some (cranium and ('bearer of' some ('color brightness' and (similar_in_magnitude_relative_to some ('color brightness' and ('inheres in' some femur))))))) and ('has part' some (cranium and ('bearer of' some ('color brightness' and (similar_in_magnitude_relative_to some ('color brightness' and ('inheres in' some ('posterior region' and ('part of' some gaster))))))))) and ('has part' some (mesosoma and ('bearer of' some ('color brightness' and (increased_in_magnitude_relative_to some ('color brightness' and ('inheres in' some cranium))))))) and )))))) and ('has part' some (mesosoma and ('bearer of' some ('color brightness' and (similar_in_magnitude_relative_to some('color brightness' and ('inheres in' some antenna))))))) and ('has part' some (mesosoma and ('bearer of' some ('color brightness' and (similar_in_magnitude_relative_to some ('color brightness' and ('inheres in' some ('anterior region' and ('part of' some gaster))))))))) and ('has part' some ((not (femur)) and ('part of' some (leg and ('bearer of' some ('color brightness' and (similar_in_magnitude_relative_to some ('color brightness' and ('inheres in' some mesosoma)))))))))Temnothorax nylanderi species-group required a complex work flow for morphometric pattern recognition. Combined stepwise analyses were done to achieve the best results. Iterated confirmatory analyses (Steps 2 and 3) helped to support the position of various samples predicted by prior steps (Step 1).The complexity of the Temnothorax nylanderi species-group. A hierarchical, agglomerative nesting method, Nest Centroid Clustering (NC clustering) was employed to reveal complex patterns in the complete dataset . \u2026Temnothorax lichtensteini - Spine length vs. absolute cephalic size (SPST/CS) < 0.384 . \u2026Temnothorax subtilis sp. n.4. Spine length vs. absolute cephalic size (SPST/CS) < 0.219 . \u2026- Spine length vs. absolute cephalic size (SPST/CS) > 0.219 \u202655. Postocular distance vs. Maximum height of the petiolar node (PoOC/NOH) > 2.678 \u20266- Postocular distance vs. Maximum height of the petiolar node (PoOC/NOH) < 2.678 \u202696. Spine length vs. absolute cephalic size (SPST/CS) < 0.283 . Turkey)\u2026- Spine length vs. absolute cephalic size (SPST/CS) > 0.283 . \u20267Temnothorax angustifrons sp. n.7. Frontal carina distance vs. absolute cephalic size (FRS/CS) < 0.334 . \u2026- Frontal carina distance vs. absolute cephalic size (FRS/CS) < 0.334 \u20268Temnothorax similis sp. n.8. Mesosoma longer ML/CS > 1.228 . Brown species. Head dorsum smooth and shiny. Ground sculpture inconspicuously areolate, smooth and shiny superimposed by feeble costulae only South Anatolia, Turkey\u2026Temnothorax helenae sp. n.- Mesosoma longer ML/CS < 1.228 . Yellow to light brown species. Whole head dorsum areolate superimposed be feeble costulae, occasionally narrow median strip inconspicuously areolate, shiny. Greek mainland, Crete and a single record known from North Anatolia, Turkey\u2026T. helenae sp. n. from T. ariadnae sp. n. may be difficult if only a single diagnostic feature is considered, more accurate option for separation is given under given in differential diagnosis under T. ariadnae sp. n.Note: Perfect separation of 9. Propodeal spines longer: SPST/CL > 0.294 \u202610- Propodeal spines shorter: SPST/CL < 0.294 \u202611Temnothorax angulinodis sp. n.10. Petiole higher: PEH/CS > 0.402 . In lateral view frontal profile of petiolar node meeting dorso-caudal plate in an acute angle (72\u201382\u00b0) with a sharp ridge. Endemic to Peloponnese peninsula, Greece\u2026Temnothorax crassispinus .- Petiole lower: PEH/CS < 0.380 . Petiolar node in lateral view with a concave frontal profile meeting truncate dorsum in an obtuse angle (100\u2013115\u00b0) with a narrowly rounded transition, without a conspicuous sharp fronto-dorsal ridge on the petiolar node. Central-East Europe and the Balkans\u2026T. crassispinus from T. crasecundus may be difficult based on a single diagnostic feature, more accurate option for their separation is given in differential diagnosis of T. crassispinus.Note: Separation of 11. Head considerably longer than broad: CL/CWb > 1.179 with straight sides\u202612- Head shorter: CL/CWb < 1.179 sides remarkably convex\u20261612. Known from mainland of Europe only\u202613- Known from Anatolian Turkey and Crete Island\u202614Temnothorax sordidulus (see note under T. tergestinus)13. Petiole longer: PL/CS > 0.422 . Black species. Main sculpture on head dorsum coarse, longitudinally rugulose and/or carinulate, ground sculpture conspicuously areolate, always dull. Endemic to Dinaran Alps\u2026Temnothorax tergestinus stat.n.- Petiole shorter: PL/CS < 0.422 . Dark brown to black species. Main sculpture on head dorsum coarse, longitudinally rugulose and/or carinulate, ground sculpture conspicuously areolate, always dull. Central and South Europe excluding Dinaran Alps\u2026T sordidulus may be difficult if only a single diagnostic feature is considered. A more accurate way for separation is suggested in\u201ddifferential diagnosis\u201d under T sordidulus.Note: Perfect separation of this species from 14. From dorsal view base of propodeal spines less distant: SPBA/CWb < 0.30 .Temnothorax lucidus sp. n.Yellow species. Head dorsum smooth and shiny. Ground sculpture inconspicuously areolate, smooth and shiny superimposed by feeble costulae only. Anatolia, Turkey\u2026- From dorsal view base of propodeal spines more distant: SPBA/CWb > 0.30 \u202615Temnothorax artvinensis Seifert, 200615. Petiolar node longer: NOL/CS > 0.251 . Black species. Main sculpture on head dorsum coarse, longitudinally rugulose and/or carinulate, ground sculpture conspicuously areolate, always dull. North Anatolia, Turkey\u2026 Temnothorax ariadnae sp. n. (see note under T. helenae sp. n.)-Petiolar node shorter: NOL/CS < 0.251 . Brown to dark brown species. Whole head dorsum uniformly areolate, narrow median strip occasionally inconspicuously areolate, shiny. Endemic to Crete\u202616. From dorsal view base of propodeal spines less distant: SPBA/CS < 0.283 \u202617- From dorsal view base of propodeal spines more distant: SPBA/CS > 0.283 \u202618Temnothorax nylanderi 17. Propodeal spines longer: SPST/CS > 0.262 . Yellow to brown species. Head dorsum with areolate ground sculpture always superimposed by parallel costulate main sculpture, dull. Central and West Europe: Italy, Austria, Germany and westward\u2026Temnothorax lucidus sp. n.- Propodeal spines shorter: SPST/CS < 0.262 . Yellow species. Head dorsum smooth and shiny. Ground sculpture inconspicuously areolate, smooth and shiny superimposed by feeble costulae only. Anatolia, Turkey\u2026Temnothorax schoedli Seifert, 200618. Postocular area shorter: PoOC/CL < 0.375 . Brown species. Main sculpture of head dorsum heterogeneous: on sides areolate ground sculpture always superimposed by parallel costulate main sculpture, dull, median part of head dorsum shiny. South-Central Anatolia, Turkey\u2026- Postocular area longer: PoOC/CL > 0.375 \u202619Temnothorax crasecundus Seifert & Cs\u0151sz, 201419. Known from East Europe, the Balkans, Turkey and the Caucasus. Yellow to brown species. Whole head dorsum areolate superimposed by feeble costulae, occasionally narrow median strip inconspicuously areolate, shiny\u2026Temnothorax nylanderi - Known from Central and West Europe: Italy, Austria, Germany and further west. Yellow to brown species. Head dorsum with areolate ground sculpture always superimposed by parallel costulate main sculpture, dull\u2026Temnothorax nylanderi species-group can be found following the diagnoses of species complexes to which they belong. Species complexes are defined based on morphological similarities, but not fully supported by molecular (mtDNA) data. Patterns formerly recognized by NC-clustering were tested by confirmatory Linear Discriminant Analysis (Steps 2 and 3). The results of complex-wise analyses (Step 3) are displayed in scatterplots for each species-complex defined below.The description and redefinition of species belonging to the Ponto-Mediterranean Temnothorax taxa , ;Paratypes: Greece_27, Peloponnesus, 22 km NNW Tripolis, Menalo Oros, 37.6539N, 22.2676E, 1500-1700mH, 29.IV.2011, leg. A. Schulz \u201e256\u201d, , [GRE:Tripolis-22NNW-20110429-256]; Greece, Peloponnesus, Melanos Oros, 20 km NNW. Tripolis, 5 km SW. Levidion, 37.6607N, 22.2546E, 1700mH, 03.06.1994, leg. A. Schulz \u201e1380\u201d, (3## HNHM), [GRE:Levidion-5SW-19940603-1380]; Greece, Peloponnesus, Melanos Oros, 20 km NNW. Tripolis, 5 km SW. Levidion, 37.6607N, 22.2546E, 1700mH, 03.06.1994, leg. A. Schulz \u201e1383\u201d (2## HNHM), [GRE:Levidion-5SW-19940603-1383];The list of 15 non-type individuals belonging to 5 nest samples of other material investigated is given in Body color: brown. Body color pattern: mesosoma, antenna and legs excluding femora, waist and anterior region of 1st gastral tergite lighter than head, femora and posterior region of gaster. Absolute cephalic size: 594\u2013657 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.171\u20131.222 (mean = 1.203). Postocular distance vs. cephalic length (PoOc/CL): 0.356\u20130.378 (mean = 0.368). Eye length vs. absolute cephalic size (EL/CS): 0.257\u20130.274 (mean = 0.265). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.366\u20130.388 (mean = 0.374). Median region of antennal rim vs. frontal carina in frontal view: not fully overlapped by frontal carina. Concentric carinae lateral to antennal foramen count: present. Carinae on medial region of frons shape: branched. Smooth median region on frons count: absent. Longitudinal carinae on median region of frons count: present. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Sculpture of submedian area of clypeus: smooth. Gena sculpture: rugoso-reticulate with feeble areolate ground sculpture. Gena frontal view shape: feebly convex. Genae contour from anterior view orientation: converging. Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular side of cranium shape: feebly convex. Vertex sculpture: main sculpture homogenously forked costate, ground sculpture areolate; main sculpture dispersed forked costate sculpture, ground sculpture areolate. Posterior margin of vertex in frontal view shape: straight. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.797\u20130.816 (mean = 0.808). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.610\u20130.636 (mean = 0.621). Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Metanotal depression count: present. Metanotal depression shape: shallow. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Spine length vs. absolute cephalic size (SPST/CS): 0.332\u20130.369 (mean = 355). Median anatomical line of propodeal spine vs. to Weber length angle value in lateral view: 32\u201338\u00b0. Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.366\u20130.398 (mean = 0.386). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.390\u20130.419 (mean = 0.409). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.262\u20130.304 (mean = 0.285). Anterodorsal rim of petiole count: present. Anterodorsal edge of petiole dorsal view shape: semicircular. Truncate dorsum of petiolar node count: absent. Truncate dorsum dorsal side contour lateral view: absent. Frontal profile of petiolar node in lateral view shape: straight. Anterodorsal edge of petiole count: present. Anterodorsal edge of petiole angle value: 72\u201382\u00b0. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed costulate. Posterodorsal edge of petiole count: absent. Caudal petiolar profile shape: straight; convex. Caudal petiolar profile angle value to ventral margin of petiole: more than 80\u00b0. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed costulate.Due to the unique combination of the long spine, high petiole, and sharp transversal crest on the dorsum of the petiolar node this species is easily distinguishable from related taxa even by simple visual inspection.This species is known only from the Peloponnese peninsula.the Temnothorax angustifrons species-complex can be distinguished from those of other complexes by the combination of the following salient features: light yellow to light brown color; moderately longer than broad head , sculpture of head dorsum shiny, with inconspicuously areolate ground sculpture combined with feeble costulate main sculpture; short to moderately long propodeal spines , deviating from longitudinal axis of mesosoma by 47\u201352\u00b0; petiolar node in lateral view with a weakly concave frontal profile meeting dorso-caudal plate in an obtuse angle (95\u2013105\u00b0) with a moderately sharp ridge, in dorsal view appearing as a visible anterior-lateral rim.Workers of Temnothorax angustifrons sp. n., T. lucidus sp. n., T. similis sp. n., T. subtilis sp. n.) within this complex, which was confirmed by LDA relative to other Temnothorax species treated in this revision.The species name refers to the frons, which is narrow , [TUR:Edremit-Kalkun-20030510-072];Paratypes: Turkey, Road Edremit to Kalkun, Kaz Da\u011fi Mountain, 39.411 N, 27.093 E, 752mH, 10.05.2003, leg. A. Schulz, \u201e072\u201d (1# HNHM), [TUR:Edremit-Kalkun-20030510-072]; Turkey, Road Edremit to Kalkun, Kaz Da\u011fi Mountain, 39.411 N, 27.093 E, 752mH, 10.05.2003, leg. A. Schulz, \u201e076\u201d (2## HNHM), [TUR:Edremit-Kalkun-20030510-076];The list of 59 non-type individuals belonging to 17 nest samples of other material investigated is given in Body color: yellow. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head dorsum and posterior region of gaster. Antennomere count: 12. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 500\u2013590 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.193\u20131.254 (mean = 1.224). Postocular distance vs. cephalic length (PoOc/CL): 0.377\u20130.401 (mean = 0.389). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture dispersed forked costate, ground sculpture inconspicuous areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with feeble areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.247\u20130.263 (mean = 0.254). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.320\u20130.349 (mean = 0.328). Longitudinal carinae on median region of frons count: present; absent. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: present. Scape length vs. absolute cephalic size (SL/CS): 0.807\u20130.832 (mean = 0.821). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 50\u201355\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.200\u20130.264 (mean = 0.231). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.242\u20130.275 (mean = 0.259). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.282\u20130.319 (mean = 0.298). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.265\u20130.298 (mean = 0.284). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.588\u20130.620 (mean = 0.601). Metanotal depression count: present. Metanotal depression shape: deep. Dorsal region of mesosoma sculpture: areolate ground sculpture, superimposed by dispersed rugae. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture dispersed costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: straight; concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 90\u2013100\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.Temnothorax angustifrons sp. n. strikingly resembles other members of this complex in Turkey. Therefore, the above specified characters may slightly overlap, but a simple ratio of their combination (FRS/SL) provides an excellent tool to separate this species from its relatives with an error rate of less than 5% for single individuals:This species has the longest scape (SL/CS) and the narrowest frons (FRS/CS) of all species treated in this revision . TemnothT. angustifrons sp.n. (n 67) = 0.398 , T. lucidus sp.n., T. similis sp.n. and T. subtilis sp.n. (n 284) = 0.461 , Combined pool of T. angustifrons sp. n. from other species belonging to other species complexes in Turkey (Simple ratios of the above-mentioned morphometric traits (SL/CS and FRS/CS) also separate n Turkey .This species is known from Western Anatolia, Turkey . Its occurn:lsid:zoobank.org:act:3C96E879-7FB4-4D94-8AC7-805CE9EF187Fm) yellow surface sculpturing of the worker caste.The species epithet \u201clucidus\u201d refers to the shiny light , [TUR:Arslank\u00f6y-3W-20111106-492];Paratypes: TUR:492 Turkey, Taurus Mt., 3 km W. Arslank\u00f6y, 37.0024 N, 34.2151 E, 1900mH, 06.11.2011, leg. A. Schulz, , [TUR:Arslank\u00f6y-3W-20111106-492]; TUR:493 Turkey, Taurus Mt., 3 km W. Arslank\u00f6y, 37.0024 N, 34.2151 E, 1900mH, 06.11.2011, leg. A. Schulz, , [TUR:Arslank\u00f6y-3W-20111106-493];The list of 65 non-type individuals belonging to 22 nest samples of other material investigated is given in Body color: yellow. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head dorsum and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 560\u2013670 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.139\u20131.221 (mean = 1.173). Postocular distance vs. cephalic length (PoOc/CL): 0.368\u20130.400 (mean = 0.380). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture dispersed forked costate, ground sculpture inconspicuous areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with feeble areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.247\u20130.276 (mean = 0.262). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.356\u20130.389 (mean = 0.370). Longitudinal carinae on median region of frons count: present; absent. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: present. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.767\u20130.832 (mean = 0.796). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 45\u201350\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.190\u20130.259 (mean = 0.236). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.241\u20130.280 (mean = 0.258). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.272\u20130.339 (mean = 0.304). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.261\u20130.318 (mean = 0.290). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.599\u20130.636 (mean = 0.621). Metanotal depression count: present. Metanotal depression shape: deep. Dorsal region of mesosoma sculpture: fine areolate ground sculpture, superimposed by dispersed rugae. Lateral region of pronotum sculpture: inconspicuous areolate ground sculpture, main sculpture dispersed costate. Mesopleuron sculpture: fine areolate ground sculpture, superimposed by dispersed rugulae. Metapleuron sculpture: fine areolate ground sculpture, superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: straight; concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 95\u2013100\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.T. angustifrons complex: T. angustifrons sp. n., T. subtilis sp. n. and T. similis sp. n. Nest samples of T. lucidus sp. n. can be separated from those of T. angustifrons sp. n. by their shorter scape (SL/CS) and wider frons (FRS/CS). Their simple ratio (FRS/SL) provides an excellent tool to separate workers with less than 5% of error rate . Ratios of NOH/CS and PEH/CS help to distinguish this species from T. similis sp. n. on the level of nest samples. A discriminant (D4) function, including discriminant scores for separating single individuals with acceptably low error rate, is given in the differential diagnosis of the latter.This species differs from members of other species complexes treated in this revision by its smooth and shiny head dorsum and the relatively short propodeal spines. This character combination is shared with other species belonging to T. lucidus sp. n. from those of T. subtilis sp. n., but in single workers this character may broadly overlap between the two species. A discriminant function with reduced character number (D4) yields 98.6% classification success rate .The spine length ratio is recommended to separate nest samples of these specie. The same function yields 97.9% classification success rate between single individuals of these species.D3 scores for single individuals:T. helenae sp. n. (n 169) = -1.982 , T. lucidus sp. n. (n 70) = +1.982 , This species is known from South and Central Anatolia, Turkey, and Crete .urn:lsid:zoobank.org:act:3BED5669-17A6-479B-9236-DA4FA22E2180m, f = similar) refers to the superficial similarity of this species to T. schoedli.Species epithet , [TUR:Antakya-22NW-20111107-512];Paratypes: TUR:512 Turkey, Nur Da\u011flari, 22 km NW. Antakya, 36.3050 N, 36.0098 E, 1600-1800mH, 07.11.2011, leg. A. Schulz, (2## HNHM), [TUR:Antakya-22NW-20111107-512]; TUR:515 Turkey, Nur Da\u011flari, 22 km NW. Antakya, 36.3050 N, 36.0098 E, 1600-1800mH, 07.11.2011, leg. A. Schulz, (4## HNHM), [TUR:Antakya-22NW-20111107-515]; TUR:520 Turkey, Nur Da\u011flari, 22 km NW. Antakya, 36.3050 N, 36.0098 E, 1600-1800mH, 07.11.2011, leg. A. Schulz, , [TUR:Antakya-22NW-20111107-520]; TUR:547 Turkey, Nur Da\u011flari, 10 km NW. Hassa, 36.8459 N, 36.4330 E, 1500mH, 08.11.2011, leg. A. Schulz, (4## HNHM), [TUR:Hassa-10NW-20111108-547]; TUR:552 Turkey, Nur Da\u011flari, 10 km NW. Hassa, 36.8459 N, 36.4330 E, 1500mH, 08.11.2011, leg. A. Schulz, , [TUR:Hassa-10NW-20111108-552];The list of 23 non-type individuals belonging to 8 nest samples of other material investigated is given in Body color: yellow; brown. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head dorsum and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 505\u2013635 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.145\u20131.233 (m = 1.183). Postocular distance vs. cephalic length (PoOc/CL): 0.371\u20130.398 (m = 0.386). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture dispersed forked costate, ground sculpture inconspicuous areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with feeble areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.245\u20130.270 (m = 0.263). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.352\u20130.369 (m = 0.361). Longitudinal carinae on median region of frons count: present; absent. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: present. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.775\u20130.830 (m = 0.802). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 10\u201315\u00b0; 35\u201340\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 47\u201352\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.220\u20130.267 (m = 0.245). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.261\u20130.290 (m = 0.277). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.299\u20130.347 (m = 0.321). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.291\u20130.326 (m = 0.321). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.606\u20130.641 (m = 0.624). Metanotal depression count: present. Metanotal depression shape: deep. Dorsal region of mesosoma sculpture: fine areolate ground sculpture, superimposed by dispersed rugae. Lateral region of pronotum sculpture: inconspicuous areolate ground sculpture, main sculpture dispersed costate. Mesopleuron sculpture: fine areolate ground sculpture, superimposed by dispersed rugulae. Metapleuron sculpture: fine areolate ground sculpture, superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 95\u2013105\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.Temnothorax similis sp. n. differs from the superficially similar T. schoedli by non-overlapping ranges of PEH/CS and NOH/CS ratios on the level of nest sample means.This species can be distinguished from other species belonging to different species complexes by its inconspicuously sculptured or smooth head and/or its shorter propodeal spines (SPST/CS). T. similis sp. n. differ from those of other species in this complex by having a wider frons (FRS/CS) than T. angustifrons sp. n., a lower petiole (NOH/CS) than T. lucidus sp. n., and longer propodeal spines (SPST/CS) than T. subtilis sp. n. is provided, that yields 98.6% classification success rate for single individuals.Though workers of . n. see , nest saD4 scores for single individuals:T. angustifrons sp.n. (n 67) = -0.732 , T. lucidus sp. n. (n 70) = -1.442, T. subtilis sp. n. (n 176) = -1.658 , T. similis sp. n. (n 38) = +1.597 , This species is known from South and Central Anatolia, Turkey . Accordiurn:lsid:zoobank.org:act:20A78840-7999-41A0-A525-CE36346A5E14The species epithet \u201esubtilis\u201d refers to the fine, tiny appearance of this species.Holotype worker labelled: TUR:431 Turkey, Taurus Mt., 5 km SW. Akseki, 37,0257 N, 31,7518 E, 950 mH, 02.11.2011, leg. A. Schulz, , [TUR:Akseki-5SW-20111102-431];Paratypes: TUR:431 Turkey, Taurus Mt., 5 km SW. Akseki, 37,0257 N, 31,7518 E, 950 mH, 02.11.2011, leg. A. Schulz, (1# HNHM), [TUR:Akseki-5SW-20111102-431]; TUR:430 Turkey, Taurus Mt., 5 km SW. Akseki, 37,0257 N, 31,7518 E, 950mH, 02.11.2011, leg. A. Schulz, (3## HNHM), [TUR:Akseki-5SW-20111102-430]; TUR:438 Turkey, Taurus Mt., 5 km SW. Akseki, 37,0257 N, 31,7518 E, 950 mH, 02.11.2011, leg. A. Schulz, (2## HNHM), [TUR:Akseki-5SW-20111102-438]; TUR:441 Turkey, Taurus Mt., 5 km SW. Akseki, 37,0257 N, 31,7518 E, 950 mH, 02.11.2011, leg. A. Schulz, (2## HNHM), [TUR:Akseki-5SW-20111102-441]; Turkey_08 Antalya, 2 km N. Imrasan Ge\u00e7idi, 12 km N. Akseki, 37,0924 N, 31,803 E, 1400mH, 03.05.1997. leg. A. Schulz, K. Vock, M. Sanetra, , [TUR:Imrasan-Ge\u00e7idi-2N-19970503-117];The list of 158 non-type individuals belonging to 50 nest samples of other material investigated is given in Body color: yellow. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head dorsum and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 499\u2013628 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.135\u20131.238 (mean = 1.189). Postocular distance vs. cephalic length (PoOc/CL): 0.374\u20130.408 (mean = 0.388). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture dispersed forked costate, ground sculpture inconspicuous areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with feeble areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.228\u20130.268 (mean = 0.249). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.335\u20130.375 (mean = 0.360). Longitudinal carinae on median region of frons count: present; absent. Smooth median region on frons count: present. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.735\u20130.810 (mean = 0.782). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 47\u201352\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.159\u20130.230 (mean = 0.192). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.247\u20130.300 (mean = 0.272). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.266\u20130.336 (mean = 0.282). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.256\u20130.322 (mean = 0.282). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.592\u20130.648 (mean = 0.623). Metanotal depression count: present. Metanotal depression shape: deep. Dorsal region of mesosoma sculpture: fine areolate ground sculpture, superimposed by dispersed rugae. Lateral region of pronotum sculpture: inconspicuous areolate ground sculpture, main sculpture dispersed costate. Mesopleuron sculpture: fine areolate ground sculpture, superimposed by dispersed rugulae. Metapleuron sculpture: fine areolate ground sculpture, superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 95\u2013105\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.T. angustifrons complex yielding 94.5% success in distinguishing nest samples .This species has the shortest propodeal spines of all species treated in this revision, and therefore can be separated from other species complexes by the non-overlapping SPST/CS ratio and its smooth and shiny head. Spine length ratio slightly overlaps with that of other species belonging to samples from T. T. subtilis sp. n. and T. similis sp. n. .A discriminant function with reduced character number (D4) arrives at 98.6% classification success between single individuals and complete success for nest sample means of T. subtilis sp. n. from those of T. lucidus sp. n., this character may broadly overlap in single individuals of these two species. In order to determine single workers with high success, a discriminant function with reduced character number (D4 = +0.0717*EL +0.0778*NOH +0.0404*SPST -0.0824*SPBA -10.321) yielding 98.6% classification success rate can be used.Though the SPST/CS ratio see providesD4 scores for single individuals:T. subtilis sp. n. (n 176) = -2.056 , T. lucidus sp. n. (n 70) = +2.098, Temnothorax subtilis sp. n. can be easily separated from two additional species of the T. parvulus complex that occur in Crete, T. ariadnae sp. n. and T. helenae sp. n., based on the shiny surface of the head dorsum. In exceptional cases or if dust cover obstructs a clear view of the surface sculpture, several ratios help to separate T. subtilis sp. n. from T. ariadnae sp. n.: it has a longer head (CL/CWb), larger eyes (EL/CS) and longer propodeal spines (SPST/CS), and a discriminant function with two characters (D2 = -0.0928*SPST +0.0215*ML -2.811) separates workers of T. subtilis sp. n. from T. helenae sp. n. if surface characteristics are not sufficient.D2 scores for single individuals:T. helenae sp. n. (n 169) = -1.760 , T. subtilis sp. n. (n 176) = 1.690 , This species is known from South Anatolia, Turkey, and Crete .Temnothorax flavicornis species-complex can be distinguished from those of other complexes treated in this revision by the combination of the following salient features: antennae 11 segmented, yellow to light brown color, head rectangular, significantly longer than broad head , sculpture of head dorsum dull: with smooth, or inconspicuously areolate ground sculpture combined with longitudinally rugulose or reticulate main sculpture; long to very long propodeal spines , deviating from longitudinal axis of mesosoma by 40\u201345\u00b0; petiolar node in lateral view with a concave frontal profile meeting conspicuously developed truncate dorsum in an obtuse angle (105\u2013115\u00b0) with a narrowly rounded transition, without a conspicuous sharp fronto-dorsal ridge on the petiolar node. This peculiar species-complex consists of a single species, Temnothorax flavicornis Emery, 1870. Separation of this species is revealed by NC-clustering corroborated by LDA ITALY.Temnothorax: Bolton, 2003: 271.Combination in Lectotype worker: \u201eLeptothorax flavicornis Em.\u201d, \u201ePortrei\u201d [\u2011] Lectotype Leptothorax flavicornis Emery, 1870 \u201eTop specimen\u201d, det. A. Schulz & M. Verhaagh 1999 (CASENT0904761) (MSNG);The list of 44 non-type individuals belonging to 12 nest samples of other material investigated is given in Body color: yellow. Body color pattern: head, mesosoma, antenna and legs excluding femora, waist and anterior region of 1st gastral tergite lighter than femora and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 465\u2013516 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.226\u20131.299 (mean = 1.266). Postocular distance vs. cephalic length (PoOc/CL): 0.370\u20130.394 (mean = 0.384). Postocular sides of cranium contour frontal view orientation: parallel. Postocular sides of cranium contour frontal view shape: straight. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture homogenously forked costate, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture; rugoso-reticulate with feeble areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.244\u20130.271 (mean = 0.262). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.351\u20130.373 (mean = 0.364). Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: absent. Antennomere count: 11. Scape length vs. absolute cephalic size (SL/CS): 0.784\u20130.817 (mean = 0.803). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 40\u201345\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.303\u20130.420 (mean = 0.358). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.322\u20130.358 (mean = 0.338). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.402\u20130.506 (mean = 0.458). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.373\u20130.482 (mean = 0.432). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.622\u20130.653 (mean = 0.637). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 105\u2013115\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: concave. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.T. flavicornis might be confused with other long-spined species, i.e., T. laconicus, T. lichtensteini and T. parvulus, but the coarse rugulose or rugulo-reticulate main sculpture on the head dorsum combined with a shiny ground sculpture help to distinguish T. flavicornis from related species by simple visual inspection. In case specimens are covered by dust, a simple ratio (SPBA/CWb) provides perfect separation of T. flavicornis and similar species at the level of nest sample means , sculpture of head dorsum dull: with areolate ground sculpture combined with longitudinally rugulose or ruguloso-reticulate main sculpture; long to very long propodeal spines , deviating from longitudinal axis of mesosoma by 20\u201325\u00b0; petiolar node in lateral view with a concave frontal profile meeting truncate dorsum in a right angle to an obtuse angle (110\u2013120\u00b0) with a narrowly rounded transition, without a conspicuous sharp fronto-dorsal ridge on the petiolar node.Workers of the Temnothorax laconicus Cs\u0151sz & al., 2014 and T. lichtensteini Bondroit, 1918) within this complex GREECE.Holotype worker: Taygetos Oros, Street to Profitis Ilias \u201eno. GRE:S_342\u201d, 36.968 N, 22.404 E, 800 mH, 01.05.2011, leg. A. Schulz (HNHM), [GRE:Profitis-Ilias-20110501-342];Paratype workers: Taygetos Oros, Street to Profitis Ilias \u201eno. GRE:S_342\u201d, 36.968 N, 22.404 E, 800 mH, 01.05.2011, leg. A. Schulz , [GRE:Profitis-Ilias-20110501-342]; W Taygetos Oros, Pigadia Canyon \u201eno. GRE:S_358\u201d, 36.984 N, 22.262 E, 700\u2013800 mH, 01.05.2011, leg. A. Schulz (2## HNHM), [GRE:Pigadia-Canyon-20110501-358]; W Taygetos Oros, Pigadia Canyon \u201eno. GRE:2011:0356\u201d 36.984 N, 22.262 E, 700\u2013800 mH, 01.05.2011, leg. A. Schulz (3## HNHM), [GRE:Pigadia-Canyon-20110501-356]; Taygetos Oros, Street to Profitis Ilias \u201eno. GRE:2011:0336\u201d, 36.968 N, 22.404 E, 800 mH, 01.05.2011, leg. A. Schulz , [GRE:Profitis-Ilias-20110501-336]; Taygetos Oros, Street to Profitis Ilias \u201eno. GRE:2011:0345\u201d 36.968 N, 22.404 E, 800 mH, 01.05.2011, leg. A. Schulz , [GRE:Profitis-Ilias-20110501-345];The list of 41 non-type individuals belonging to 10 nest samples of other material investigated is given in Body color: brown. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 500\u2013590 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.199\u20131.258 (mean = 1.228). Postocular distance vs. cephalic length (PoOc/CL): 0.383\u20130.403 (mean = 0.396). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: strongly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture homogenously forked costate, ground sculpture areolate; main sculpture absent, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.229\u20130.265 (mean = 0.244). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.329\u20130.360 (mean = 0.342). Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.766\u20130.804 (mean = 0.785). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0; 35\u201345\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 20\u201325\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.391\u20130.429 (mean = 0.411). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.257\u20130.311 (mean = 0.283). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.401\u20130.485 (mean = 0.438). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.381\u20130.462 (mean = 0.413). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.587\u20130.629 (mean = 0.610). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 110\u2013120\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.Temnothorax laconicus can be distinguished easily from other species by its very long propodeal spines and its low deviation (20\u201325\u00b0) from the mesosomal axis.T. lichtensteini. The simple ratio SPST/CS does not overlap between the two species at the level of nest sample means : -0.656 , T. laconicus sp.n. (n 64) +3.085 , This species is known to occur in the Peloponnese peninsula and Kerkira .Leptothorax lichtensteini Bondroit, 1918: 123 (w.q.m.) FRANCE.Temnothorax: Bolton, 2003: 271.Combination in Lectotype and paralectotypes: 4 workers labeled \"Montpellier Jean Lichtenstein\", \"Leptoth. lichtensteini Type Bondr.\" and \"Lectotype Leptothorax lichtensteini Bondroit, 1918 Top specimen det. A.Schulz & M.Verhaagh 1999\"; IRSNB Bruxelles; lectotype with CS 546.6. 4 workers labeled \"Menton de Dalmas\" and \"Leptoth. lichtensteini Type Bondr.\"; IRSNB Bruxelles. (4## IRSNB), [FRA:lichtensteini-type:montpellier];The list of 298 non-type individuals belonging to 84 nest samples of other material investigated is given in Body color: brown. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 474\u2013585 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.181\u20131.261 (mean = 1.225). Postocular distance vs. cephalic length (PoOc/CL): 0.386\u20130.418 (mean = 0.401). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: strongly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture homogenously forked costate, ground sculpture areolate; main sculpture absent, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.232\u20130.270 (mean = 0.248). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.336\u20130.380 (mean = 0.356). Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.763\u20130.809 (mean = 0.787). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 35\u201345\u00b0; 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 20\u201325\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.324\u20130377 (mean = 0.346). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.250\u20130.302 (mean = 0.272). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.335\u20130.428 (mean = 0.391). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.318\u20130.411 (mean = 0.371). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.570\u20130.631 (mean = 0.608). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 110\u2013120\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: concave. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.Temnothorax lichtensteini complex (T. lichtensteini and T. laconicus) can be easily distinguished from other species treated in this revision by the very long propodeal spines and their low deviation (20\u201325\u00b0) from the mesosomal axis. Other species with long spines have more erect propodeal spines deviating from the mesosomal axis by >35\u00b0. How T. lichtensteini and T. laconicus can be separated is described under the latter (see above).Members of the Temnothorax lichtensteini is distributed throughout the Northern coast of the Mediterranean basin from Spain to Turkey and \u201cWest Mediterranean cluster\u201d ;The list of 167 non-type individuals belonging to 56 nest samples of other material investigated is given in Body color: yellow; brown. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head dorsum and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 539\u2013719 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.121\u20131.196 (mean = 1.155). Postocular distance vs. cephalic length (PoOc/CL): 0.378\u20130.405 (mean = 0.374). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: strongly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture dispersed forked costate sculpture, ground sculpture areolate; main sculpture parallel costate, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex; convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.236\u20130.280 (mean = 0.255). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.359\u20130.397 (mean = 0.374). Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: not forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.763\u20130.836 (mean = 0.791). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 32\u201335\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.253\u20130.322 (mean = 0.289). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.279\u20130.321 (mean = 0.298). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.311\u20130.392 (mean = 0.350). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.300\u20130.369 (mean = 0.331). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.612\u20130.662 (MEAN = 0.631). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 88\u2013104\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: slightly convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.T. crassispinus. The D4 function that separates these two species is given under the latter.This species is easily confused with its parapatric relative, Temnothorax crasecundus shares most of its surface sculpturing and general shape characteristics with T. helenae sp. n., despite the fact that they belong to different species complexes based on molecular phylogeny and morphometrics. In addition, the distribution of these species broadly overlaps in Bulgaria, Greece, and Turkey. A simple ratio (PoOC/NOH) yields a rather reliable discrimination with minor overlap between them (T. crasecundus is larger (CS), has a wider frons (FRS/CS), higher petiolar node (NOH/CS), and longer propodeal spines (SPST/SC) than T. helenae sp. n. UKRAINE.Temnothorax: Bolton, 2003: 271.Combination in Temnothorax slavonicus: Radchenko, 2000: 44.Senior synonym of Leptothorax nylanderi var. crassispina Karavajev, 1926:Syntype workers of Golossev near Kiev, Leg. Karawajew, \"No. 3057 Col. Karawajew\" [UKR:Golossev-crassispinus-TYPE];Leptothorax nylanderi slavonicus Seifert, 1995:Paratype workers of Germany, Kr. G\u00f6rlitz, Hutberg Sch\u00f6nau-Berzdorf, 19.03.1993. Seifert (4## SMNG) [GER:Hutberg-slavonicus-TYPE];The list of 119 non-type individuals belonging to 40 nest samples of other material investigated is given in Body color: brown; yellow. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 544\u2013688 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.100\u20131.180 (mean = 1.140). Postocular distance vs. cephalic length (PoOc/CL): 0.379\u20130.403 (mean = 0.390). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: strongly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture parallel costate, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.247\u20130.268 (mean = 0.256). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.362\u20130.399 (mean = 0.377). Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: not forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.756\u20130.811 (mean = 0.784). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 32\u201342\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.288\u20130.356 (mean = 0.329). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.282\u20130.339 (mean = 0.312). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.362\u20130.421 (mean = 0.389). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.342\u20130.397 (mean = 0.366). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.509\u20130.662 (mean = 0.626). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 100\u2013115\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: slightly convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.Temnothorax crassispinus may be confused with other long-spined species treated in this revision: T. angulinodis sp.n., T. laconicus, T. lichtensteini and T. parvulus. Temnothorax angulinodis sp.n. clearly differs from T. crassispinus by its sharply angulate petiolar node in lateral view (72\u201382\u00b0). In T. crassispinus, the frontal profile and the truncate dorsum of the petiole meet in an obtuse angle (100\u2013115\u00b0). The deviation of the propodeal spines from longitudinal mesosomal axis helps to separate T. crassispinus (32\u201342\u00b0) from T. laconicus and T. lichtensteini (20\u201325\u00b0). Temnothorax parvulus differs from T. crassispinus in the surface sculpturing on the head dorsum. If the samples are dust-covered, other measures can also help: T. crassispinus is considerably larger (CS), has a higher petiolar node (SPST/CS), and a wider head (CL/CWb) than T. parvulus helps to separate nest samples of T. crassispinus from those of T. nylanderi without error, but the same character overlaps between nest sample means of T. crassispinus and T. crasecundus. The shortest discriminant formula (D4) that separates T. crassispinus from T. crasecundus with a classification success rate 95% in single individuals and 97% in nest sample means is D4 = +0.0392*SL -0.0746*SPST +0.0933*SPL -0.0295*SPWI -7.2179.D4 scores for single individuals:T. crassispinus (n 139) = -1.458 , T. crasecundus (n 161) = +1.458 , D4 scores for nest sample means:T. crassispinus (n 45) = -1.440 , T. crasecundus (n 54) = +1.479 , Temnothorax crassispinus lays between the ranges of its two parapatric relatives, T. nylanderi in the West and T. crasecundus in the East.This species is distributed from the Balkans to Central Europe . The disMyrmica nylanderi Foerster, 1850: 53 (m.) GERMANY.Temnothorax: Bolton, 2003: 271.Combination in Leptothorax nylanderi Foerster, 1852Type material of is not available and most probably lost. Altogether 6 workers belonging to two nest series from the type locality \u201eAachen\u201d were investigated: Germany, vic. Aachen-Brand, 5km SE Aachen, 50.7506 N, 6.1202 E, 200 mH, 21.06.1999. leg. A. Schulz, , [GER:Aachen-5SE-19990621-1]; and , [GER:Aachen-5SE-19990621-2] with the same label data;The list of 60 non-type individuals belonging to 20 nest samples of other material investigated is given in Body color: brown; yellow. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head dorsum and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 587\u2013678 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.121\u20131.160 (mean = 1.140). Postocular distance vs. cephalic length (PoOc/CL): 0.379\u20130.402 (mean = 0.391). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: strongly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture parallel costate, ground sculpture areolate; main sculpture absent, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.245\u20130.268 (mean = 0.254). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.354\u20130.383 (mean = 0.373). Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: not forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.757\u20130.798 (mean = 0.777). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 35\u201342\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.265\u20130.297 (mean = 0.280). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.263\u20130.294 (mean = 0.280). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.321\u20130.362 (mean = 0.343). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.298\u20130.339 (mean = 0.320). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.610\u20130.646 (mean = 0.624). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 100\u2013115\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: slightly convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.Temnothorax nylanderi has moderately long spines (SPST/CS) and therefore cannot be confused with long-spined T. lichtensteini. Non-overlapping SPBA/CS ratios help to distinguish it from T. flavicornis reliably separate these species on the level of nest sample means , sculpture of head dorsum dull: with uniformly areolate ground sculpture combined with inconspicuous (or the lack of) main sculpture; short to long propodeal spines , deviating from longitudinal axis of mesosoma by 38\u201342\u00b0; petiolar node in lateral view with a concave frontal profile meeting truncate dorsum in a right angle to an obtuse angle (100\u2013110\u00b0) with a narrowly rounded transition, without a conspicuous sharp fronto-dorsal ridge on the petiolar node.Workers of the Temnothorax ariadnae sp. n., T. helenae sp. n., and T. parvulus , which was corroborated by confirmatory analyses , [GRE:Crete-Ano-Vianos-5N-20110417-024];Paratypes: GER:024 Greece, Crete, 5 km N Ano Vianos, Vic. Katofigi, 35.0922 N, 25.4165 E, 60 mH, 17.04.2011, leg. A. Schulz, , [GRE:Crete-Ano-Vianos-5N-20110417-024]; GRE:092 Greece, Crete, 3 km E Ag. Vasilios, 25 km S Rethimnon, 35.2408 N, 24.4652 E, 300mH, 24.04.2011, leg. A. Schulz, (5## HNHM), [GRE:Crete-Ag-Vasilios-3E-20110424-092]; GRE:093 Greece, Crete, 3 km E Ag. Vasilios, 25 km S Rethimnon, 35.2408 N, 24.4652 E, 300mH, 24.04.2011, leg. A. Schulz, , [GRE:Crete-Ag-Vasilios-3E-20110424-093]; GRE:037 Greece, Crete, Lassithi Plateau, 16 km S Malia, 35.1623 N, 25.4560 E, 1000mH, 18.04.2011, leg.A. Schulz, , [GRE:Crete-Malia16S-20110418-037];Locality data of the single non-type sample with 3 individuals investigated is given in Body color: brown; yellow. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head dorsum and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 533\u2013557 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.196\u20131.293 (mean = 1.223). Postocular distance vs. cephalic length (PoOc/CL): 0.373\u20130.393 (mean = 0.386). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: strongly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture absent, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.260\u20130.266 (mean = 0.263). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.346\u20130.369 (mean = 0.356). Longitudinal carinae on median region of frons count: absent. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.758\u20130.795 (mean = 0.775). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0; 35\u201345\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 45\u201350\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.220\u20130.249 (mean = 0.237). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.248\u20130.287 (mean = 0.278). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.303\u20130.336 (mean = 0.314). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.289\u20130.322 (mean = 0.300). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.602\u20130.618 (mean = 0.611). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: strongly convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.T. helenae sp. n. and T. parvulus. Values of the propodeal spine length ratio (SPST/CL) for nest sample means perfectly separate T. parvulus and T. ariadnae sp. n. . The geographical range of T. ariadnae sp. n. and T. parvulus function is the shortest formula that yields reliable separation for single individuals (99.4%) and nest sample means (100%) of T. ariadnae sp. n. and T. helenae sp. n.The separation of D4 scores for single individuals:T. ariadnae sp. n. (n 15) = +1.607 , T. helenae sp. n. (n 169) = -1.607 , D4 scores for nest sample means:T. ariadnae sp.n. (n 5) = +1.607 T. helenae sp.n. (n 53) = -1.678 Temnothorax ariadnae sp. n. can be easily separated from two additional species whose distributional range expands to Crete, T. lucidus sp. n. and T. subtilis sp. n., based on the shiny surface of the head dorsum of the two latter. In exceptional cases, or if dust cover obstructs a clear view of the surface sculpture, body ratios help to distinguish T. ariadnae sp. n. from T. subtilis sp. n. by the longer head (CL/CWb), the larger eyes (EL) and the longer propodeal spines (SPST/CS) and from T. lucidus sp. n. by the non-overlapping NodL/CS, lower NOL/CS ratio and the longer head , [GRE:Taigetos-Oros-20110501-344];Paratypes: GRE:344 Greece, Peloponnesus, Taygetos Oros, Street to Profitis Elias, 36.968 N, 22.404 E, 800mH, 01.05.2011, leg. A. Schulz, (6## HNHM), [GRE:Taigetos-Oros-20110501-344]; GRE:267 Greece, Peloponnesus, Taygetos Oros, Trail to Profitis Elias, 36.960 N, 22.396 E, 1000-1200mH, 30.04.2011, leg. A. Schulz, , [GRE:Taigetos-Oros-20110430-267]; GRE:280 Greece, Peloponnesus, Taygetos Oros, Trail to Profitis Elias, 36.960 N, 22.396 E, 1000-1200mH, 30.04.2011, leg. A. Schulz, (5## HNHM), [GRE:Taigetos-Oros-20110430-280]; GRE:287 Greece, Peloponnesus, Taygetos Oros, Trail to Profitis Elias, 36.948 N, 22.377 E, 1400-1600mH, 30.04.2011, leg. A. Schulz, , [GRE:Taigetos-Oros-20110430-287]; GRE:348 Greece, Peloponnesus, Taygetos Oros, Street to Profitis Elias, 36.968 N, 22.404 E, 800mH, 01.05.2011, leg. A. Schulz, , [GRE:Taigetos-Oros-20110501-348]; GRE:350 Greece, Peloponnesus, Taygetos Oros, Street to Profitis Elias, 36.968 N, 22.404 E, 800mH, 01.05.2011, leg. A. Schulz, (5## HNHM), [GRE:Taigetos-Oros-20110501-350];The list of 154 individuals belonging to 43 nest samples of other material investigated is given in Body color: brown; yellow. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head dorsum and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 510\u2013627 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.152\u20131.242 (mean = 1.197). Postocular distance vs. cephalic length (PoOc/CL): 0.384\u20130.424 (mean = 0.400). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: feebly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture absent, ground sculpture areolate; main sculpture dispersed forked costate sculpture, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.238\u20130.270 (mean = 0.251). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.335\u20130.373 (mean = 0.357). Longitudinal carinae on median region of frons count: absent; present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: present; absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.758\u20130.808 (mean = 0.783). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 45\u201350\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.205\u20130.299 (mean = 0.255). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.255\u20130.319 (mean = 0.281). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.290\u20130.387 (mean = 0.334). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.277\u20130.370 (mean = 0.320). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.580\u20130.634 (mean = 0.611). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 100\u2013110\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: slightly convex to conspicuously rounded. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.Temnothorax helenae sp. n. shares characters with T. ariadnae sp. n. and T. parvulus, differential diagnoses between these species are given under these taxa.T. helenae sp. n. from T. crasecundus can be difficult if only sculpture and shape characteristics are considered, but a single ratio (PoOC/NOH) yields >95% classification success for nest sample means function yields 97% classification success for individuals and 100% for nest sample means.The geographical distribution of D5 scores for nest sample means:T. helenae sp. n. (n = 53) = -2.209 T. lucidus sp. n. (n = 23) = +1.410 T. subtilis sp. n. (n = 55) = +1.579 This species mostly occurs in Greek mainland, but a few localities are also known in Southern Bulgaria, Western Turkey and Crete .Myrmica parvula Schenck, 1852: 103 (w.) GERMANY. PalearcticTemnothorax: Bolton, 2003: 271.Combination in Lectotype: \"L. parvulus # Sch\" and \"Lectotype Leptothorax parvulus Schenck, 1852 det. A. Schulz & M. Verhaagh 1999\" (1# / SMF), [GER:parvulus-TYPE];The list of 103 individuals belonging to 27 nest samples of other material investigated is given in Body color: brown; yellow. Body color pattern: mesosoma, antenna and legs, waist and anterior region of 1st gastral tergite lighter than head dorsum and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 488\u2013586 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.147\u20131.214 (mean = 1.184). Postocular distance vs. cephalic length (PoOc/CL): 0.392\u20130.413 (mean = 0.405). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: strongly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture absent, ground sculpture areolate; main sculpture dispersed forked costate sculpture, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.237\u20130.262 (mean = 0.250). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.353\u20130.376 (mean = 0.361). Longitudinal carinae on median region of frons count: absent. Smooth median region on frons count: present. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.763\u20130.796 (mean = 0.778). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 38\u201342\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.278\u20130.331 (mean = 0.306). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.273\u20130.312 (mean = 0.292). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.353\u20130.415 (mean = 0.384). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.332\u20130.395 (mean = 0.364). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.599\u20130.636 (mean = 0.618). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 105\u2013110\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: slightly convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.Temnothorax parvulus can be separated easily from members of other species-complexes by its homogenously areolate microsculpture on the head dorsum. The head sculpture of other species complexes may vary from smooth to coarsely rugulo-reticulate but is never homogenously areolate. In exceptional cases, long-spined species of the T. lichtensteini complex might exhibit a homogenous areolate sculpture on an extended area of the head dorsum, but T. parvulus is distinguished from T. lichtensteini and T. laconicus by its more erect propodeal spines (38\u201342\u00b0 vs. ca. 20\u201325\u00b0). Temnothorax parvulus can also be safely separated from weakly sculptured, lightly colored T. tergestinus samples using slightly overlapping NOH/CS, SPTI/CS and SPWI/CS ratios helps distinguish Temnothorax parvulus from T. ariadnae sp. n. and T. helenae sp. n.T. parvulus ; and T. ariadnae sp. n. provide a prefect means of separation. As 5% of nest samples of T. parvulus and T. helenae sp. n. formula, which separates 97.1% of single individuals and all nest samples of T. parvulus and T. helenae sp. n.Non-overlapping values of SPST/CL for nest sample means of D5 scores for single individuals:T. parvulus (n 107) = -1.624 T. helenae sp.n. (n 169) = +1.647 D5 scores for nest sample means:T. parvulus (n 29) = -1.723 T. helenae sp.n. (n 53) = +1.684 The known distribution of this species ranges from Western Europe to the Black see coast and Turkey and from Italy and the Balkans to Central Europe .Temnothorax sordidulus species-complex can be distinguished from those of other complexes treated in this revision by the combination of the following salient features: brown to black color; slightly longer than broad head ], sculpture of head dorsum dull: with areolate ground sculpture combined with conspicuous parallel costulate or irregular reticulate main sculpture; moderately long to long propodeal spines , deviating from longitudinal axis of mesosoma by 40\u201350\u00b0; petiolar node in lateral view with a concave frontal profile meeting occasionally indistinct truncate dorsum in an obtuse angle (110\u2013120\u00b0) with a narrowly rounded transition, without a conspicuous sharp fronto-dorsal ridge on the petiolar node. Four species consist of this complex: Temnothorax artvinensis Seifert, 2006, T. schoedli Seifert, 2006, T. sordidulus and T. tergestinus stat.n.Workers of the Temnothorax angustifrons sp. n., T. lucidus sp. n., T. similis sp. n., T. subtilis sp. n.) within this complex, which was confirmed by LDA TURKEY.Paratypes: 9 paratype workers of 3 nest samples were investigated from the type locality: Turkey, Artvin, 5 km SW. Artvin, 41.1445 N, 41.8537 E, 1000 mH, 27.06.1993, leg. A. Schulz \u201eno. 1162\u201d (3## ASPC) [TUR:Artvin-5SW-19930627-1162]; \u201eno. 1164\u201d (3## ASPC) [TUR:Artvin-5SW-19930627-1164]; \u201eno. 1165\u201d (3## ASPC) [TUR:Artvin-5SW-19930627-1165];The list of 39 individuals belonging to 12 nest samples of other material investigated is given in Body color: brown; black. Body color pattern: head, mesosoma, waist and anterior region of 1st gastral tergite lighter than antenna and legs and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 540\u2013634 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.206\u20131.263 (mean = 1.240). Postocular distance vs. cephalic length (PoOc/CL): 0.375\u20130.389 (0.384). Postocular sides of cranium contour frontal view orientation: parallel; converging posteriorly. Postocular sides of cranium contour frontal view shape: feebly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture homogenously forked costate, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.239\u20130.260 (mean = 0.247). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.357\u20130.375 (mean = 0.367). Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.776\u20130.813 (mean = 0.794). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: carinate. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 42\u201348\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.227\u20130.288 (mean = 0.265). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.265\u20130.301 (mean = 0.278). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.296\u20130.350 (mean = 0.331). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.280\u20130.337 (mean = 0.313). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.598\u20130.626 (mean = 0.612). Metanotal depression count: present. Metanotal depression shape: deep. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 110\u2013120\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: slightly convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: concave; straight. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.T. artvinensis prevents confusion of this species with any other Turkish Temnothorax treated in this revision.The very dark color of T. artvinensis is geographically well isolated from other dark brown or black species of the T. sordidulus complex, it is unlikely that this this species is confused with its siblings. However, for doubtful cases, or if anthropochory is suspected, a discriminant function after character reduction (D4 = +0.114*EL -0.124*PEW +0.098*NOH -0.068*PPL +3.69209) separates 97.9% of T. artvinensis workers and each nest sample from that of Turkish T. schoedli and European brownish-black T. sordidulus and T. tergestinus.Due to the fact that D4 scores for nest sample means:T. artvinensis (n 15) = -1.620 T. schoedli (n 11) = +2.044 T. tergestinus (n 49) = +1.575 T. sordidulus (n 15) = +2.218 This species occurs in a restricted area of North-East Turkish high mountains .Temnothorax schoedli Seifert, 2006: 8 (w.q.) TURKEY.Paratypes: 20 paratype workers of 7 nest samples were investigated from the type locality: Turkey, Antakya, Nur Da\u011flari, 14 rkm W. Hassa 36.8414 N, 36.4309 E, 1600 mH, 11.05.1997, leg. A. Schulz, K. Vock, M. Sanetra: \u201eno. 287\u201d (2## SMNK) [TUR:Hassa-14W-19970511-287]; \u201eno. 289\u201d (3## SMNK) [TUR:Hassa-14W-19970511-289]; \u201eno. 290\u201d (3## SMNK) [TUR:Hassa-14W-19970511-290]; \u201eno. 292\u201d (3## ASPC) [TUR:Hassa-14W-19970511-292]; \u201eno. 293\u201d (3## ASPC) [TUR:Hassa-14W-19970511-293]; \u201eno. 295\u201d (3## ASPC) [TUR:Hassa-14W-19970511-295]; \u201eno. 299\u201d (3## ASPC) [TUR:Hassa-14W-19970511-299];The list of 12 non-type individuals belonging to 4 nest samples of other material investigated is given in Body color: brown. Body color pattern: mesosoma, antenna and legs excluding femora, waist and anterior region of 1st gastral tergite lighter than head, femora and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 572\u2013696 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.143\u20131.196 (mean = 1.172). Postocular distance vs. cephalic length (PoOc/CL): 0.360\u20130.385 (mean = 0.368). Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: broadly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture dispersed forked costate, ground sculpture inconspicuous areolate; main sculpture dispersed forked costate sculpture, ground sculpture areolate; main sculpture homogenously forked costate, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.255\u20130.282 (mean = 0.268). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.363\u20130.379 (mean = 0.372). Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.764\u20130.805 (mean = 0.784). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 40\u201345\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.232\u20130.282 (mean = 0.261). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.282\u20130.298 (mean = 0.288). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.305\u20130.342 (mean = 0.327). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.293\u20130.328 (mean = 0.310). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.613\u20130.650 (mean = 0.632). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: areolate ground sculpture, superimposed by dispersed rugae. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture dispersed costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: widely rounded or slightly angulate area. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.T. sordidulus species-complex, Temnothorax schoedli has the lightest color. In addition to color it can be separated safely from other members of the complex by its very restricted geographical occurrence in the southern ranges of Taurus Mountains in Turkey. Temnothorax schoedli can be separated from T. artvinensis, which in geographical terms is the most closely situated relative of this species, by the coarse dull head sculpture and the very dark brown to black color of the latter and its significantly shorter head. The non-overlapping CL/CWb ratio ; from T. similis sp. n. by PEH/CS and NOH/CS ratios; from T. lucidus sp. n. by SPBA/CS; and from T. angustifrons sp. n. by the FRS/CS ratio.In its relatively lighter color and occasionally smooth head dorsum this species may superficially resemble species of g ratios : T. schoThis species occurs in a restricted area of South-Eastern Turkish lowlands .Leptothorax sordidulus M\u00fcller, 1923: 96 (w.) ITALY.Temnothorax: Bolton, 2003: 271.Combination in Temnothorax carinthiacus: Schulz, 1991: 121.Senior synonym of Leptothorax sordidulus M\u00fcller, 1923:Type material of not investigated, most probably lost, see also Seifert (2006).Leptothorax carinthiacus Bernard, 1957:Lectotype and Paralectotype of Carinthia Viktring H\u00f6lzler leg. [on reverse side: \u201eV/55\u201d], Lectotype Leptothorax carinthiacus Bernard 1957 desig. B. Seifert 2006 (2# LMK), [AUT:Viktring-carinthiacus-TYPE] (CASENT0913637);The list of 43 individuals belonging to 14 nest samples of other material investigated is given in Body color: brown; black. Body color pattern: head, mesosoma, waist and anterior region of 1st gastral tergite lighter than antenna and legs and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 518\u2013607\u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.192\u20131.278 (mean = 1.238). Postocular distance vs. cephalic length (PoOc/CL): 0.363\u20130.390 (mean = 0.374). Postocular sides of cranium contour frontal view orientation: parallel; converging posteriorly. Postocular sides of cranium contour frontal view shape: feebly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture homogenously forked costate, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.258\u20130.280 (mean = 0.266). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.353\u20130.378 (mean = 0.366). Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.763\u20130.809 (mean = 0.785). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0. Ground sculpture of submedian area of clypeus: smooth; carinate. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 42\u201348\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.227\u20130.281 (mean = 0.258). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.257\u20130.291 (mean = 0.273). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.280\u20130.331 (mean = 0.306). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.266\u20130.312 (mean = 0.288). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.599\u20130.640 (mean = 0.615). Metanotal depression count: present. Metanotal depression shape: deep. Dorsal region of mesosoma sculpture: rugulose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture forked costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 110\u2013120\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: slightly convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.T. tergestinus. The other black species belonging to this complex, T. artvinensis is endemic to Turkey. Though T. sordidulus seems endemic to the Dinaran Alps, its distribution slightly overlaps with that of T. tergestinus in Slovenia and Austria. The darker, often black color of T. sordidulus in contrast to the brown color of Central European populations of T. tergestinus, its shorter spines (SPST/CS), shorter scapes (SL/CS), longer petioles (PL/CS) and petiolar nodes (NOL/CS) may help to separate T. sordidulus from Central European populations of T. tergestinus. If these ratios overlap and do not clearly separate between the two taxa, or if T. sordidulus shall be separated from Bulgarian or Greek populations of T. tergestinus, a discriminant function (D7 = +0.0323*SPTI -0.0594*NOL +0.0859*SL -0.0490*ML +0.0867*CWb -0.1167*EL -0.0414*CL -1.2164) yields 99% classification success rate in single individuals and 100% in nest sample means.Due to its dark brown to black color this species can only be confused with its European sibling, D7 scores for single individuals:T. sordidulus (n 47) = -1.872 , T. tergestinus (n 139) = +1.872 , D7 scores for nest sample means:T. sordidulus (n 16) = -1.885 T. tergestinus (n 49) = +1.868 T. sordidulus cannot be confused with any other species treated in this revision.In its restricted distributional area This species has a relatively restricted distributional area and is occurs in the Dinaran Alps of Austria, Croatia and Slovenia .Leptothorax sordidulus var. tergestina Finzi, 1928: 129 (w) ITALY.Temnothorax: Bolton, 2003: 271.Combination in Temnothorax saxonicus : [synonymy proposed hereby]Senior synonym of Leptothorax sordidulus var. tergestina Finzi, 1928:Syntype workers of \u201eS.Croze\u201d Ven. Giulia [Trieste] B. Finzi \u201e6.27\u201d, FinziColl. purch 1950, M. C. Z. \u201eco\u201dtype \u201e28840\u201d, \u201eSyntypus Leptothorax sordidulus var tergestinus Finzi\u201d, [on the reverse side: \u201eSP Cover 98\u201d], MCZ Museum of Comparative Zoology (4## MCZ), [ITA:Ven-Giulia-tergestinus-TYPE];Leptothorax sordidulus saxonixus Seifert, 1995:Nest sample of the holotype of GER: 51.0895 N, 14.6927 E L\u00f6bauer Berg 415m Basaltklippen mit Quercus, B. Seifert 1983.07.28\u2013473 aus Holotypus-Nest (3## HNHM), [GER:Lobauer-Berg-saxonicus-TYPE];The list of 132 non-type individuals belonging to 46 nest samples of other material investigated is given in Body color: brown. Body color pattern: head, mesosoma, waist and anterior region of 1st gastral tergite lighter antenna and legs except femora lighter than femora and posterior region of gaster. Antenna color pattern: clava concolorous funicle. Absolute cephalic size: 523\u2013665 \u03bcm . Cephalic length vs. Maximum width of head capsule (CL/CWb): 1.169\u20131.253 (mean = 1.206). Postocular distance vs. cephalic length (PoOc/CL): 0.373\u20130.400 (mean = 0.384). Postocular sides of cranium contour frontal view orientation: parallel; converging posteriorly. Postocular sides of cranium contour frontal view shape: broadly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture homogenously forked costate, ground sculpture areolate. Genae contour from anterior view orientation: converging. Gena contour line in frontal view shape: feebly convex. Gena sculpture: rugoso-reticulate with areolate ground sculpture. Median region of antennal rim vs. frontal carina in frontal view structure: not fully overlapped by frontal carina. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.237\u20130.280 (mean = 0.258). Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.347\u20130.380 (mean = 0.361). Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.781\u20130.824 (mean = 0.799). Facial area of the scape absolute setal angle: 0\u201315\u00b0. External area of the scape absolute setal angle: 30\u00b0; 35\u201345\u00b0. Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 45\u201350\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.220\u20130.335 (mean = 0.276). Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.248\u20130.295 (mean = 0.273). Maximum spine distance vs. absolute cephalic size (SPWI/CS): 0.283\u20130.372 (mean = 0.333). Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.270\u20130.346 (mean = 0.312). Maximum mesosoma width vs. absolute cephalic size (MW/CS): 0.588\u20130.652 (mean = 0.620). Metanotal depression count: present. Metanotal depression shape: shallow. Dorsal region of mesosoma sculpture: areolate ground sculpture, superimposed by dispersed rugae. Lateral region of pronotum sculpture: areolate ground sculpture, main sculpture dispersed costate. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Metapleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae. Frontal profile of petiolar node contour line in lateral view shape: concave. Dorsal profile of petiolar node contour line angle value to frontal profile of petiole contour line in lateral view: 110\u2013120\u00b0. Anterodorsal rim of petiole count: absent medially. Dorsal profile of petiolar node contour line in lateral view shape: slightly convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Dorso-caudal petiolar profile contour line in lateral view shape: straight; concave. Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent.T. parvulus and T. nylanderi, particularly if the body surface is covered by diffuse dust.This species can be separated from members of other species complexes by the ruguloreticulate main sculpture on head dorsum that turns irregular on the vertex and the sides of the head. In some Western and Central European populations, the surface sculpturing might be less conspicuous, which may lead to possible confusion with T. tergestinus can be safely separated from T. nylanderi using PoOC/CL and non-overlapping SPWI/CS ratios . A discriminant function (D7) that helps separating T. tergestinus from T. sordidulus is given in differential diagnosis under the latter.on range and by dT. sordidulus and T. saxonicus . Our results, however, differ from the earlier [T. sordidulus, but grouped in a cluster that was formerly referred to as T. saxonicus is nested in the T. saxonicus fell in the same cluster with a posterior probability of p = 0.99. Therefore, we propose a new junior synonymy for Temnothorax saxonicus with Temnothorax tergestinus stat. n.A multivariate analyses of 22 continuous characters in 186 individuals of 64 nest samples and 29 c earlier view in Seifert ); b) in Seifert ) clusterT. tergestinus and T. sordidulus , in contrast to the 18 characters that was used to require earlier [The newly outlined species boundaries allow a considerably easier separation of lus Figs and 29, earlier .T. tergestinus spreads from France to Bulgaria and Greece .All samples can be uniquely distinguished by sample-specific abbreviations Click here for additional data file.S2 TableTemnothorax nylanderi species-complex.GenBank accession number of CO I sequences of ants of (FAS)Click here for additional data file.S3 TableTemnothorax nylanderi species-complex. Data are given in \u03bcm.Morphometric characters of worker individuals of (CSV)Click here for additional data file.S4 TableTemnothorax nylanderi species-complex treated in this revision. Mean of indices, \u00b1SD are provided in the upper row, minimum and maximum values are given in parentheses in the lower row.Nest sample means of continuous morphometric data are calculated for the eighteen species of (XLSX)Click here for additional data file.S5 TableTemnothorax nylanderi species-complex investigated in this revisionary work is shown.Identification matrix calculated by Linear Discriminant Analysis and Leave One Out Cross Validation for 1693 workers of (XLSX)Click here for additional data file.S6 TableTemnothorax nylanderi species-group is provided with their semantic representations in Manchester Syntax format.The list of natural language phenotypes used in the species descriptions of (XLSX)Click here for additional data file."} +{"text": "Sensors [http://www.mdpi.com/1424-8220/15/3/5251.The authors wish to update the Acknowledgments in their paper published in Sensors , doi:10."} +{"text": "The second author\u2019s last name is misspelled. The correct name is: Alexis C. Frazier-Wood. The correct citation is given below.Cheung CHM, Frazier-Wood AC, Asherson P, Rijsdijk F, Kuntsi J (2014) Shared Cognitive Impairments and Aetiology in ADHD Symptoms and Reading Difficulties. PLoS ONE 9(6): e98590. doi:10.1371/journal.pone.0098590"} +{"text": "PLoS ONE 9(3): e91694. doi:10.1371/journal.pone.0091694The second author\u2019s name is incorrect. The correct name is Robin Fortt. The correct citation is: Witney TH, Fortt R, Aboagye EO (2014) Preclinical Assessment of Carboplatin Treatment Efficacy in Lung Cancer by"} +{"text": "The fourth author\u2019s name is spelled incorrectly. The correct name is: Alex Lu. The correct citation is: Debruin EJ, Hughes MR, Sina C, Lu A, Cait J, et al. (2014) Podocalyxin Regulates Murine Lung Vascular Permeability by Altering Endothelial Cell Adhesion. PLoS ONE 9(10): e108881. doi:10.1371/journal.pone.0108881"} +{"text": "Dr. Yusuke Naito is the third author and also a corresponding author for this article. His affiliation is:2Sanofi-aventis K.K., Tokyo, Japan. His contributions are as follows: analyzed the data and wrote the paper.The correct citation and contact information of the corresponding authors are:Kadowaki T, Ohtani T, Naito Y, Odawara M (2012) Potential Formula for the Calculation of Starting and Incremental Insulin Glargine Doses: ALOHA Subanalysis. PLoS ONE 7(8): e41358. doi:10.1371/journal.pone.0041358Corresponding Authorskadowaki-3im@h.u-tokyo.ac.jp)Takashi Kadowaki (yusuke.naito@sanofi.com)Yusuke Naito ("} +{"text": "There are errors in the Author Contributions. The correct contributions are: Conceived and designed the experiments: AC AJW A. Mondino AAM PRQ. Performed the experiments: AC GC ER VB MV A. Monno. Analyzed the data: AC GF ER AJW A. Mondino AAM PRQ. Contributed reagents/materials/analysis tools: AJW A. Mondino AAM PRQ. Wrote the paper: AC GF AEA A. Mondino AJW AAM PRQ.The following information is missing from the Funding section: AIRC\u2014Associazione Italiana per la Ricerca sul Cancro (IG 11970/2011 to A.M.).The publisher apologizes for these errors."} +{"text": "The third author\u2019s name is spelled incorrectly. The correct name is Eric Vatikiotis-Bateson. The correct citation is: Latif N, Barbosa AV, Vatikiotis-Bateson E, Castelhano MS, Munhall KG (2014) Movement Coordination during Conversation. PLoS ONE 9(8): e105036. doi:10.1371/journal.pone.0105036"} +{"text": "Opisthorchis felineus Acquired in Italy , author Wouter Rozemeijer\u2019s name was spelled incorrectly. The article has been corrected online (http://wwwnc.cdc.gov/eid/article/20/1/13-0476_article).In the article Foodborne Trematodiasis and"} +{"text": "The fifth author\u2019s name is spelled incorrectly. The correct name is: Justin I. Odegaard. The correct citation is: Richardson ML, Fu C-L, Pennington LF, Honeycutt JD, Odegaard JI, et al. (2014) A New Mouse Model for Female Genital Schistosomiasis. PLoS Negl Trop Dis 8(5): e2825. doi:10.1371/journal.pntd.0002825"} +{"text": "Recent tastases . Preclintastases -5. As a tastases to a protastases . Paralletastases . In humatastases . Dranofflearance . Decreaslearance . Since Rlearance biopsies"} +{"text": "Arabidopsis thaliana Using TAL-Effector-Nucleases. PLoS ONE 10(3): e0121056. doi:10.1371/journal.pone.0121056The fourth author\u2019s name is spelled incorrectly. The correct name is: Pablo A. Manavella. The correct citation is: Forner J, Pfeiffer A, Langenecker T, Manavella PA, Lohmann JU (2015) Germline-Transmitted Genome Editing in"} +{"text": "The second author\u2019s name is spelled incorrectly. The correct name is: \u015e. Utku Yavuz. The correct citation is: Negro F, Yavuz \u015eU, Farina D (2014) Limitations of the Spike-Triggered Averaging for Estimating Motor Unit Twitch Force: A Theoretical Analysis. PLoS ONE 9(3): e92390. doi:10.1371/journal.pone.0092390"} +{"text": "P. aureofaciens LMG 2145 should be P. aureofaciens 1245. P. amygdali LMG1384 should be P. amygdali 13184. P. resinovorans LMG 2774 should be P. resinovorans LMG 2274. P. mandelii LMG 2210 should be P. mandelii LMG 21607. P. luteola LMG 21607 should be P. luteola LMG 7041. Please see the corrected Figs There are errors in the names of type strains appearing in Figs S1 FiggyrB, rpoD and rpoB genes from 451 sequenced genomes and 107 type strains (bold), ML method and Tamura-Nei model. C. japonicus Ueda 107 was used as outgroup. Only bootstrap values above 75% over 1000 replicates are shown. Bold and T indicates type strain.MLSA based on partial sequences of 16S rDNA, (PDF)Click here for additional data file."} +{"text": "Aging (Albany NY) 2015; 7(2): 97-109.PMCID: PMC4359692 PMID: 25701668http://www.impactaging.com/papers/v7/n2/pdf/100718.pdf.In this Article, the error was found in Figure 5: The correct Figure is provided here."} +{"text": "Cryptococcus gattii Populations, Vancouver Island, Canada, 2002\u20132004 . The article has been corrected online (http://wwwnc.cdc.gov/eid/article/21/11/14-1161_article).Cryptococcus was misspelled in the title of Climatic Influences on"} +{"text": "There are errors in the Author Contributions. The correct contributions are: Conceived and designed the experiments: HO VT SS RL MNP LK JA. Performed the experiments: HO VT MW. Analyzed the data: HO VT SS RL MNP LK JA. Contributed reagents/materials/analysis tools: RL MNP LK JA. Wrote the paper: HO VT."} +{"text": "Following the publication of the original article , it was \u201820. Freire SM, Sundvall E, Karlsson D, Lambrix P. Performance of XML Databases for Epidemiological Queries in Archetype-Based EHRs. 2012:51\u201357.\u2019However, it should instead read as:http://www.ep.liu.se/ecp/070/009/ecp1270009.pdf\u2019\u201820. Freire SM, Sundvall E, Karlsson D, Lambrix P. Performance of XML Databases for Epidemiological Queries in Archetype-Based EHRs. Scandinavian Conference on Health Informatics 2012:51\u201357."} +{"text": "The matrix for FTIR spectra is available under the DOI: http://dx.doi.org/10.6084/m9.figshare.1300167. The report of S/G ratio in thirteen Cactaceae species studie by HPLC is available under the DOI: http://dx.doi.org/10.6084/m9.figshare.1300168. Supporting Information of the S/G ratio in thirteen Cactaceae species studied by HPLC is available under the DOI: http://dx.doi.org/10.6084/m9.figshare.1300165. Supporting Information of the percentage of syringyl-size plant graphic is available under the DOI: http://dx.doi.org/10.6084/m9.figshare.1300166.The Data Availability statement for this paper is incorrect. The correct statement is: All relevant data are available from Figshare. The complete dataset of chemical composition in Cactaceae wood is available under the DOI:"} +{"text": "Nucl. Acids Res. 43 (12): e80. doi: 10.1093/nar/gkv242Johan den Dunnen's middle initial was omitted. The author's full name should be Johan T. den Dunnen.The publisher apologises to the authors for this error."} +{"text": "Aurelia Species: Understanding Jellyfish Ecology Using Genetics and Morphometrics. PLoS ONE 11(6): e0156588. doi:10.1371/journal.pone.0156588The second author's name is spelled incorrectly. The correct name is: Keith M. Bayha. The correct citation is: Chiaverano LM, Bayha KM, Graham WM (2016) Local versus Generalized Phenotypes in Two Sympatric"} +{"text": "AbstractNesomyrmex is reviewed and (1) subdivided into four major groups based on salient morphological features corroborated by numeric morphology: angulatus-, hafahafa-, madecassus- and sikorai-groups, and (2) the hafahafa species-group endemic to Madagascar is revised. Diversity within hafahafa species-group was assessed via hypothesis-free nest-centroid-clustering combined with gap statistic to assess the number of clusters and to determine the most probable boundaries between them. This combination of methods provides a highly automatized, objective species delineation protocol based on continuous morphometric data. Delimitations of clusters recognized by these exploratory analyses were tested via confirmatory Linear Discriminant Analysis. These results suggest the existence of four morphologically distinct species, Nesomyrmexcapricornissp. n., Nesomyrmexhafahafasp. n., Nesomyrmexmedusussp. n. and Nesomyrmexspinosussp. n.; all are described and an identification key for their worker castes using morphometric data is provided. Two members of the newly outlined hafahafa species-group, Nesomyrmexhafahafasp. n., Nesomyrmexmedusussp. n., are distributed along the southeastern coast Madagascar and occupy rather large ranges, but two other species, Nesomyrmexcapricornissp. n. and Nesomyrmexspinosussp. n., are only known to occur in small and isolated forest, highlighting the importance of small forest patches for conserving arthropod diversity.Madagascar and its surrounding islands are among the world\u2019s greatest biodiversity hotspots, harboring predominantly endemic and threatened communities meriting special attention from biodiversity scientists. Building on the considerable efforts in recent years to inventory the Malagasy ant fauna, the myrmicine genus The Malagasy zoogeographical region, i.e. Madagascar and surrounding islands , is consNesomyrmex have never been the subject of focused research. To date, only four valid Nesomyrmex species have been recorded to occur in Madagascar makes the appearance of this group extremely unique; no similar species group has been found either in the Malagasy region or on the African continent. Multivariate evaluation of morphological data has revealed that the unique-looking Nesomyrmexhafahafa species-group comprises four well-outlined clusters, or species, that are endemic to Madagascar. The four new species outlined, Nesomyrmexcapricornis sp. n., Nesomyrmexhafahafa sp. n., Nesomyrmexmedusus sp. n., and Nesomyrmexspinosus sp. n., are described here based on worker caste, and both a key that includes both a numeric identification tool that helps readers to resolve the most problematic cases and a traditional character based key. Distribution maps are also provided. Our research PageBreakhas also revealed that two of the four species, Nesomyrmexcapricornis sp. n. and Nesomyrmexspinosus sp. n., occur in small, highly isolated forests, leaving them at a high risk of extinction from continuing environmental destruction or climatic changes.In the present paper, the Malagasy California Academy of Sciences (CAS), San Francisco, USA. The full list of non-type material morphometrically examined in this revision is listed in Table CASENT0460666). Designation of type material with detailed label information is given in relevant sections type material investigated for each taxon. All images and specimens used in this study are available online on AntWeb (http://www.antweb.org). Images are linked to their specimens via their unique specimen code affixed to each pin (CASENT0002660). Online specimen identifiers follow this format: http://www.antweb.org/specimen/CASENT0002660.In the present study, 21 continuous morphometric traits were recorded in 177 worker individuals belonging to 100 nest samples collected in the Malagasy region , or a Leica DFC 425 camera in combination with the Leica Application Suite software (version 3.8). Distribution maps were generated by using QGIS 2.4.0 software .The measurements were taken with a Leica MZ 12.5 stereomicroscope equipped with an ocular micrometer at a magnification of 100\u00d7. Measurements and indices are presented as arithmetic means with minimum and maximum values in parentheses. Body size dimensions are expressed in \u00b5m. Due to the abundance of worker individuals in contrast to the limited number of queen and male specimens available the present revision is based on worker caste only. Worker-based revision is further facilitated by the fact that name-bearing type specimens of the vast majority of existing ant taxa were designated from worker caste. All measurements were made by the first author. For the definition of morphometric characters, earlier protocols were con Maximum cephalic length in median lineCL. The head must be carefully tilted to the position providing the true maximum. Excavations of hind vertex and/or clypeus reduce CL Mesosoma length from caudalmost point of propodeal lobe to transition point between anterior pronotal slope and anterior pronotal shieldML . In gynes: length from caudalmost point of propodeal lobe to the most distant point of steep anterior pronotal face phenotypes were compiled in mx (http://purl.org/NET/mx-database). Taxonomic history and descriptions of taxonomic treatments were rendered from this software. Hymenoptera-specific terminology of morphological statements used in descriptions, identification key, and diagnoses are mapped to classes in phenotype-relevant ontologies (Hymenoptera Anatomy Ontology (HAO) via a URgy (HAO) ; see SelIn verbal descriptions of taxa based on external morphological traits, recent taxonomic papers were conHypothesis formation by exploratory analyses. Our hypothesis of the number of clusters and classification of samples was formulated by an exploratory data analysis technique, NC-clustering ( (Unweighted Pair Group Method with Arithmetic Mean)UPGMA distance method. This method is able to tackle large datasets with high dimensionality is computed by multiscale bootstrap resampling, and the raw Bootstrap Probabilities (BP) that is calculated before statistical adjustments by normal bootstrap resampling.ustering using coionality , providi were given to samples that were incongruently classified by the two methods. The confirmative LDA was run as an iterative process to achieve the lowest number of characters necessary to achieve the desired level (>97%) of classification success angulatus ilgii = latinodis = angulatus concolor = hafahafa groupcapricornis Cs\u0151sz & Fisher,sp. n.hafahafa Cs\u0151sz & Fisher,sp. n.medusus Cs\u0151sz & Fisher,sp. n.spinosus Cs\u0151sz & Fisher,sp. n.PageBreakmadecassus groupgibber madecassus sikorai groupretusispinosus sikorai Absolute cephalic size (CS): 591 \u00b5m . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.218 . Postocular distance vs. cephalic length (PoOc/CL): 0.40 . Scape length vs. absolute cephalic size (SL/CS): 0.676 . Eye length vs. absolute cephalic size (EL/CS): 0.260 . Petiole width vs. absolute cephalic size (PEW/CS): 0.431 . Postpetiole width vs. absolute cephalic size (PPW/CS): 0.496 . Petiolar node height vs. absolute cephalic size (PEW/CS): 0.250 . Nesomyrmexangulatus and ca. four undescribed species belong to this group in the Malagasy zoogeographical region.Pronotal spines present or absent. Anterodorsal spines on petiolar node absent. Propodeal spines short to long and acute. Vertex ground sculpture areolate. Main sculpture on vertex not defined. Metanotal depression present or absent. Median clypeal notch present or absent. Median clypeal notch shape/depth: 0\u201323 \u00b5m. Antennomere count: 12. PageBreakPageBreakCS): 1059 \u00b5m . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.074 . Postocular distance vs. cephalic length (PoOc/CL): 0.378 . Scape length vs. absolute cephalic size (SL/CS): 0.890 . Eye length vs. absolute cephalic size (EL/CS): 0.232 . Petiole width vs. absolute cephalic size (PEW/CS): 0.267 . Postpetiole width vs. absolute cephalic size (PPW/CS): 0.523 . Petiolar node height vs. absolute cephalic size (PEW/CS): 0.142 . Four species, Nesomyrmexcapricornis sp. n., Nesomyrmexhafahafa sp. n., Nesomyrmexmedusus sp. n. and Nesomyrmexspinosus sp. n. are known to constitute this species group in Madagascar.Pronotal spines present. Anterodorsal spines on petiolar node present. Propodeal spines long and acute. Vertex ground sculpture areolate. Vertex main sculpture rugulose. metanotal depression absent. Median clypeal notch present. Median clypeal notch shape/depth: 15\u201331 \u00b5m. Antennomere count: 12. Absolute cephalic size (CS): 571 \u00b5m . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.231 . Postocular distance vs. cephalic length (PoOc/CL): 0.479 . Scape length vs. absolute cephalic size (SL/CS): 0.718 . Eye length vs. absolute cephalic size (EL/CS): 0.249 . Petiole width vs. absolute cephalic size (PEW/CS): 0.217 . Postpetiole width vs. absolute cephalic size (PPW/CS): 0.331 . Petiolar node height vs. absolute cephalic size (PEW/CS): 0.122 . Nesomyrmexmadecassus and ca. seven other taxa from the Malagasy zoogeographical region will be revised in the forthcoming revisionary work.Pronotal spines absent. Anterodorsal spines on petiolar node absent. Propodeal spines short, lamelliform to absent. Vertex ground sculpture smooth. Vertex main sculpture not defined. Metanotal depression present. Median clypeal notch present or absent. Median clypeal notch shape/depth 0\u201315 \u00b5m. Antennomere count: 12. Absolute cephalic size (CS): 750 \u00b5m . Cephalic length vs. maximum width of head capsule (CL/CWb): 1.218 . Postocular distance vs. cephalic length (PoOc/CL): 0.461 . Scape length vs. absolute cephalic size (SL/CS): PageBreak0.816 . Eye length vs. absolute cephalic size (EL/CS): 0.232 . Petiole width vs. absolute cephalic size (PEW/CS): 0.243 . Postpetiole width vs. absolute cephalic size (PPW/CS): 0.359 . Petiolar node height vs. absolute cephalic size (PEW/CS): 0.175 . Nesomyrmexsikorai , Nesomyrmexretusispinosus plus ca. ten more Malagasy species will be revised in a forthcoming revisionary work.Pronotal spines present or absent. Anterodorsal spines on petiolar node absent. Propodeal spines short to long and acute. Vertex ground sculpture not defined. Vertex main sculpture areolate. Metanotal depression present. Median clypeal notch present or absent. Median clypeal notch shape/depth 0\u201315 \u00b5m. Antennomere count: 12. Absolute cephalic size +(0.0121\u00d7SPST)-(0.0023\u00d7PSTI)+(0.0281\u00d7NSTI) +1.6LD1= -(0.0324\u00d7PEL)+(0.0258\u00d7SPST)-(0.0328\u00d7PSTI)+(0.0049\u00d7NSTI)-2.9LD2= + obtained here can either be compared to the values given in Table Though all species defined in this revisionary work proved to be highly separable via descriptive morphology, or by using simple indices, the application of classification functions LD1 and LD2 provides a foolproof, numeric morphology-based identification tool when decisions based on conventional diagnostic traits fail.PageBreakNesomyrmexhafahafa species-group are described, and a key to these species is provided. Diagnoses are given in the key, the basic statistics of body size ratios are given in Table hafahafa group is detailed in the discussion. The diagnoses and a key to the four Malagasy Nesomyrmex species groups defined here are followed by the descriptions of species belonging to the hafahafa group.In this section, four new species of the PageBreakNesomyrmexhafahafa group differ in body ratios. The following dichotomous identification key for the worker caste was generated based on ratios of morphological features that allow quick identification. Minimum and maximum values for each character is given in parentheses. The reliability of all characters has been tested and calculated classification success was always higher than 95% for each node. Where classification error was detected (i.e. the range of a given trait overlaps between two species) a percentile range 5\u201395% was also provided in brackets.The species of the Taxon classificationAnimaliaHymenopteraFormicidaeCs\u0151sz & Fishersp. n.http://zoobank.org/EC84BA51-2D96-4084-AB2B-8B19AF1DEEDCHolotype worker.CASENT0452741, collection code: BLF05245; MADGAGASCAR: Prov. Toliara, For\u00eat Mahavelo, Isantoria Riv., 5.2 km 44\u00b0NE Ifotaka, 24\u00b046'S, 46\u00b009'E , 110 m, 28.iii.2002 Fisher et al. (CAS);Paratypes. Ten workers, a single gyne and two males with the same label data with the holotype under CASENT codes: CASENT0452715, \u201c5245\u201d, ; CASENT0452716, \u201c5245\u201d, ; CASENT0452717, \u201c5245\u201d, ; CASENT0452720, BLF05245, ; CASENT0452721, BLF05245, ; CASENT0452722, BLF05245, ; CASENT0452725, BLF05245, ; CASENT0452726, BLF05245, ; CASENT0452726, BLF05245, ; CASENT0452727, BLF05245, ; CASENT0452728, BLF05245, ; CASENT0452729, BLF05245, ; CASENT0452730, BLF05245, ; CASENT0452731, BLF05245, ; CASENT0452732, BLF05245, ; CASENT0452733, BLF05245, ; CASENT0452734, BLF05245, ; CASENT0452735, BLF05245, ; CASENT0452736, BLF05245, ; CASENT0452737, BLF05245, ; CASENT0452738, BLF05245, ; CASENT0452739, BLF05245, ; CASENT0452742, BLF05245, ; CASENT0452743, BLF05245, ; CASENT0452744, BLF05245, ; CASENT0452745, BLF05245, ; CASENT0452746, BLF05245, ; CASENT0452747, BLF05245, ; CASENT0452748, BLF05245, ; CASENT0452750, BLF05245, ; CASENT0452751, BLF05245, ; CASENT0452752, BLF05245, ; CASENT0452753, BLF05245, ;The list of 21 non-type individuals belonging to 14 nest samples of other material investigated is given in Table In key.CL/CWb): 1.079 . Postocular distance vs. cephalic length (PoOc/CL): 0.390 . Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: broadly convex. Vertex contour line in frontal view shape: straight. Vertex sculpture: main sculpture rugose, ground sculpture areolate. Gena contour line in frontal view shape: convex. Genae contour from anterior view orientation: converging. Gena sculpture: rugo-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen count: absent; present. Eye length vs. absolute cephalic size (EL/CS): 0.241 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.315 . Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.927 . Facial area of the scape absolute setal angle: setae absent, pubescence only. Median clypeal notch count: present. Median clypeal notch depth vs. absolute cephalic size (Cdep/CS): 0.023 . Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 65\u201370\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.397 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.260 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.455 . Propodeal spine shape: straight; slightly bent. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.658 . Metanotal depression count: absent. Dorsal region of mesosoma sculpture: areolate ground sculpture, superimposed by dispersed rugae. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Metapleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.265 . Anterodorsal spines on petiolar node angle of deviation from each other: 60\u00b0. Apical distance of anterodorsal spines on petiolar node vs. absolute cephalic size (NSTI/CS): 0.265 . Frontal profile of petiolar node contour line in lateral view shape: straight; concave. Dorso-caudal petiolar profile contour line in lateral view shape: strongly convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.558 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture absent; ground sculpture areolate, main sculpture dispersed rugose.Body color: yellow. Body color pattern: Body concolorous, only clava darker. Absolute cephalic size: 1024 \u00b5m (n=27). Cephalic length vs. maximum width of head capsule in the southern part of Madagascar Fig. .Taxon classificationAnimaliaHymenopteraFormicidaeCs\u0151sz & Fishersp. n.http://zoobank.org/C2249F7A-0FFE-4C76-A2E8-905A4B1EA754This Malagasy word \u201chafahafa\u201d means weird, and refers to the unusual morphology of this species.Holotype worker.CASENT0460666, collection code: BLF06010; MADG\u2019R: Prov. Toliara, For\u00eat de Tsinjoriaky, 6.2 km 84\u00b0 E Tsifota, 22\u00b048'S, 43\u00b025'E , 70 m, 6\u201310.iii.2002 Fisher et al. (CAS)Paratypes. Ten workers, a single gyne and two males with the same label data as the holotype under CASENT codes: CASENT0746771, BLF06010, ; CASENT0460667, BLF06010, ; CASENT0460668, BLF06010, ; CASENT0460669, BLF06010, ; CASENT0451364, \u201c6019\u201d, ; CASENT0451364, \u201c6019\u201d, ;The list of 44 non-type individuals belonging to 25 nest samples of other material investigated is given in Table In key.CL/CWb): 1.224 [1.193-1.254]. Postocular distance vs. cephalic length (PoOc/CL): 0.388 . Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: broadly convex. Vertex contour line in frontal view shape: straight; slightly concave. Vertex sculpture: main sculpture PageBreakrugose, ground sculpture areolate. Gena contour line in frontal view shape: feebly convex. Genae contour from anterior view orientation: converging. Gena sculpture: rugo-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.230 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.316 . Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.895 . Facial area of the scape absolute setal angle: setae absent, pubescence only. Median clypeal notch count: present. Median clypeal notch depth vs. absolute cephalic size (Cdep/CS): 0.022 . Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine anPageBreakgle value to Weber length in lateral view: 55\u201360\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.398 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.287 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.543 . Propodeal spine shape: strongly bent. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.724 . Metanotal depression count: absent. Dorsal region of mesosoma sculpture: areolate ground sculpture, superimposed by dispersed rugae. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture superimposed by dispersed rugulae; areolate ground sculpture, superimposed by dispersed rugae. Metapleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.307 . Anterodorsal spines on petiolar node angle of deviation from each other: 80\u00b0. Apical distance of anterodorsal spines on petiolar node vs. absolute cephalic size (NSTI/CS): 0.514 . Frontal profile of petiolar node contour line in lateral view shape: convex. Dorso-caudal petiolar profile contour line in lateral view shape: convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.538 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture dispersed rugose.Body color: yellow; brown. Body color pattern: body concolorous, only clava darker. Absolute cephalic size: 1062 \u00b5m (n = 48). Cephalic length vs. maximum width of head capsule ;Paratypes. Ten workers, a single gyne and two males with the same label data as the holotype under CASENT codes: CASENT0746770, BLF06201, ; CASENT0455429, BLF06201, ; CASENT0455430, BLF06201, ; CASENT0455431, BLF06201, ; CASENT0455432, BLF06201, ; CASENT0455433, BLF06201, ; CASENT0455434, BLF06201, ; CASENT0455435, BLF06201, ; CASENT0455437, BLF06201, ; CASENT0455438, BLF06201, ; CASENT0455439, BLF06201, ; CASENT0455440, BLF06201, ;The list of 54 non-type individuals belonging to 28 nest samples of other material investigated is given in Table In key.CL/CWb): 1.046 . Postocular distance vs. cephalic length (PoOc/CL): 0.391 . Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: broadly convex. Vertex contour line in frontal view shape: straight; slightly concave. Vertex sculpture: main sculpture rugose, ground sculpture areolate. Gena contour line in frontal view shape: feebly convex. Genae contour from anterior view orientation: converging. Gena sculpture: rugo-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.232 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.313 . Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.907 . Facial area of the scape absolute setal angle: setae absent, pubescence only. Median clypeal notch count: present. Median clypeal notch depth vs. absolute cephalic size (Cdep/CS): 0.022 . Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 65\u201372\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.385 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.266 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.443 . Propodeal spine shape: straight; slightly bent. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.757 . Metanotal depression count: absent. Dorsal region of mesosoma sculpture: areolate ground sculpture, superimposed by dispersed rugae. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Metapleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.268 . Anterodorsal spines on petiolar node angle of deviation from each other: 70\u00b0. Apical distance of anterodorsal spines on petiolar node vs. absolute cephalic size (NSTI/CS): 0.354 . Frontal profile of petiolar node contour line in lateral view shape: straight. Dorso-caudal petiolar profile contour line in lateral view shape: straight; convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture dispersed rugose; ground sculpture areolate, main sculpture absent. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.543 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture absent; ground sculpture areolate, main sculpture dispersed rugose.Body color: brown. Body color pattern: body concolorous, only clava darker. Absolute cephalic size: 1069 \u00b5m (n=56). Cephalic length vs. maximum width of head capsule of Madagascar Fig. between Taxon classificationAnimaliaHymenopteraFormicidaeCs\u0151sz & Fishersp. n.http://zoobank.org/D3643DB1-75EB-415A-9220-9F255A5FCB21Name \u201cspinosus\u201d refers to the short, strong antero-dorsal spines on the petiolar node.Holotype worker.CASENT0443515, BLF05489; MADGAGASCAR: Prov. Toliara, R\u00e9serve Priv\u00e9 Berenty, For\u00eat d\u2019Anjapolo, 21.4 km 325\u00b0 NW Amboasary, 24\u00b056'S, 46\u00b013'E , 65 m, 7.iii.2002 Fisher et al. (CASCASENT0443515);Paratypes. 24 workers and three males with the same label data with the holotype under CASENT codes: CASENT0443515, BLF05489, ; PageBreakCASENT0443516, BLF05489, ; CASENT0443517, BLF05489, ; CASENT0443518, BLF05489, ; CASENT0443519, BLF05489, ; CASENT0443520, BLF05489, ; CASENT0443521, BLF05489, ; CASENT0443522, BLF05489, ; CASENT0443523, BLF05489, ; CASENT0443524, BLF05489, ; CASENT0443525, BLF05489, ; CASENT0443526, BLF05489, ; CASENT0443527, BLF05489, ; CASENT0443530, BLF05489, ; CASENT0443531, BLF05489, ; CASENT0443532, BLF05489, ; CASENT0443533, BLF05489, ; CASENT0443534, BLF05489, ; CASENT0443535, BLF05489, ; CASENT0443536, BLF05489, ; CASENT0443537, BLF05489, ;The list of 44 non-type individuals belonging to 26 nest samples of other material investigated is given in Table In key.PageBreakCL/CWb): 1.056 . Postocular distance vs. cephalic length (PoOc/CL): 0.374 . Postocular sides of cranium contour frontal view orientation: converging posteriorly. Postocular sides of cranium contour frontal view shape: broadly convex. Vertex contour line in frontal view shape: slightly concave. Vertex sculpture: main sculpture rugose, ground sculpture areolate. Gena contour line in frontal view shape: feebly convex. Genae contour from anterior view orientation: converging. Gena sculpture: rugo-reticulate with areolate ground sculpture. Concentric carinae laterally surrounding antennal foramen count: present. Eye length vs. absolute cephalic size (EL/CS): 0.239 . Frontal carina distance vs. absolute cephalic size (FRS/CS): 0.315 . Longitudinal carinae on median region of frons count: present. Longitudinal carinae on medial region of frons shape: forked. Smooth median region on frons count: absent. Antennomere count: 12. Scape length vs. absolute cephalic size (SL/CS): 0.880 . Facial area of the scape absolute setal angle: setae absent, pubescence only. Median clypeal notch count: present. Median clypeal notch depth vs. absolute cephalic size (Cdep/CS): 0.021 . Ground sculpture of submedian area of clypeus: smooth. Median carina of clypeus count: present. Lateral carinae of clypeus count: present. Median anatomical line of propodeal spine angle value to Weber length in lateral view: 65\u00b0. Spine length vs. absolute cephalic size (SPST/CS): 0.300 . Minimum spine distance vs. absolute cephalic size (SPBA/CS): 0.212 . Apical spine distance vs. absolute cephalic size (SPTI/CS): 0.307 . Propodeal spine shape: straight; slightly bent. Apical distance of pronotal spines vs. absolute cephalic size (PSTI/CS): 0.677 . Metanotal depression count: absent. Dorsal region of mesosoma sculpture: rugose with areolate ground sculpture. Lateral region of pronotum sculpture: areolate ground sculpture, superimposed by dispersed rugae. Mesopleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Metapleuron sculpture: areolate ground sculpture, superimposed by dispersed rugae. Petiole width vs. absolute cephalic size (PEW/CS): 0.237 . Anterodorsal spines on petiolar node angle of deviation from each other: 60\u00b0. Apical distance of anterodorsal spines on petiolar node vs. absolute cephalic size (NSTI/CS): 0.216 . Frontal profile of petiolar node contour line in lateral view shape: straight. Dorso-caudal petiolar profile contour line in lateral view shape: convex. Dorsal region of petiole sculpture: ground sculpture areolate, main sculpture absent; ground sculpture areolate, main sculpture dispersed rugose. Postpetiole width vs. absolute cephalic size (PPW/CS): 0.491 . Dorsal region of postpetiole sculpture: ground sculpture areolate, main sculpture absent; ground sculpture areolate, main sculpture dispersed rugose.Body color: brown. Body color pattern: body concolorous, only clava darker. Absolute cephalic size: 1021 \u00b5m (n=46). Cephalic length vs. maximum width of head capsule in the southern part of Madagascar Fig. .PageBreakNesomyrmex fauna into four species-groups delimited based on morphological features corroborated by morphometric data (see definition and diagnoses of groups). The within-group diversity of one of these new groups, Nesomyrmexhafahafa group, was revealed by an enhanced hypothesis-free approach. The exploratory NC-clustering implemented in the package clusterGenomics (part\u2019) assigned observations into partitions. Gap statistic is a global method, determines the number of clusters based on gap criterion described by A gap statistic algorithm and the known biogeographic patterns (Fig. Our research demonstrates that combination of NC-clustering with gap statistics and recursive partitioning algorithms performs well in distinguishing partitions in the present data based on morphological distances among nest sample means. Four-cluster hypothesis was returned by both gap statistic Fig. and recurns Fig. .Nesomyrmex.We highlight the importance and advantages of the combination of NC-clustering with algorithms to statistically infer gaps and create array of clusters. This protocol also has the potential at accelerate and improve taxonomic decision making process considerably by enabling taxonomists to objectively interpret results based on quantitative morphometric data even in a largely underexplored or poorly understood group such as the Malagasy genus Combination of these approaches allows researchers to recognize cryptic species, but also prevent users from inferring overly diverse pattern in the data. A taxonomist without long-term training in a given group can evaluate new specimens and potential new species by repeating the analysis with measurements from new specimens. This method is best included with an integrated approach that includes conventional morphological characters, biogeography, ecology or molecular data.PageBreak"} +{"text": "The third author\u2019s name is spelled incorrectly. The correct name is: Robert Peu\u00df. The correct citation is: Milutinovi\u0107 B, Stolpe C, Peu\u00df R, Armitage SAO, Kurtz J (2013) The Red Flour Beetle as a Model for Bacterial Oral Infections. PLoS ONE 8(5): e64638. doi:10.1371/journal.pone.0064638"} +{"text": "Inorg. DOI: 10.1107/S1600536814013336/hb7216Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S1600536814013336/hb7216Isup3.cdxSupporting information file. DOI: 1007160CCDC reference: crystallographic information; 3D view; checkCIF reportAdditional supporting information:"} +{"text": "Tel: +254 20 418 32 39www.kcco.net or contact Genes Mng'anga at genes@kcco.netVisit Contact Anita Shah admin@cehjournal.orgweb@cehjournal.org or visit www.cehjournal.org/subscribewww.cehjournal.orgwww.facebook.com/CEHJournal/https://twitter.com/CEHJournalVisit us online:"} +{"text": "The second author\u2019s name is incorrect. The correct name is Willscott Edward Naugler. The correct citation is: Ellis ECS, Naugler WE, Parini P, M\u00f6rk L-M, Jorns C, et al. (2013) Mice with Chimeric Livers Are an Improved Model for Human Lipoprotein Metabolism. PLoS ONE 8(11): e78550. doi:10.1371/journal.pone.0078550"} +{"text": "From 2001 to 2011, the percentage of uninsured persons aged <65 years for whom cost was a reason for not having health insurance coverage decreased among uninsured Hispanic, non-Hispanic white, and non-Hispanic black persons. In 2001 and 2011, uninsured Hispanic persons aged <65 years were more likely than uninsured non-Hispanic white and non-Hispanic black persons to lack health insurance coverage because of cost.Sources: Barnes PM, Adams PF, Schiller JS. Summary health statistics for the U.S. population: National Health Interview Survey, 2001. Vital Health Stat 2003;10(217). Available at http://www.cdc.gov/nchs/data/series/sr_10/sr10_217.pdf.http://www.cdc.gov/nchs/data/series/sr_10/sr10_255.pdf.Adams PF, Kirzinger WK, Martinez ME. Summary health statistics for the U.S. population: National Health Interview Survey, 2011. Vital Health Stat 2012;10(255). Available at"} +{"text": "The first author\u2019s name is spelled incorrectly. The correct name is: Julian Klein. The correct citation is: Klein J, Darvin ME, M\u00fcller KE, Lademann J (2012) Serial Non-Invasive Measurements of Dermal Carotenoid Concentrations in Dairy Cows following Recovery from Abomasal Displacement. PLoS ONE 7(10): e47706. doi:10.1371/journal.pone.0047706"} +{"text": "Phytophthora plurivora. PLoS ONE 9(1): e85368. doi:10.1371/journal.pone.0085368The third author\u2019s name is spelled incorrectly. The correct name is: Niklaus J. Gr\u00fcnwald. The correct citation is: Schoebel CN, Stewart J, Gr\u00fcnwald NJ, Rigling D, Prospero S (2014) Population History and Pathways of Spread of the Plant Pathogen"} +{"text": "Phil. Trans. R. Soc. A\u2009380, 20210299. (Published online 15 August 2022). (https://doi.org/10.1098/rsta.2021.0299)In the original version of this article, references 113\u2013120, 123\u2013140 and 143 were incorrectly numbered.This has been corrected on the publisher's website."} +{"text": "This article has been corrected: In 37154-37163. https://doi.org/10.18632/oncotarget.16209Original article: Oncotarget. 2017; 8:37154\u201337163."} +{"text": "Correction: Psicologia: Reflex\u00e3o e Cr\u00edtica 35, 35 (2022)https://doi.org/10.1186/s41155-022-00237-9BLINDED\u201d in the citation should be changed to the Kinkead et al., 2021.Following publication of this article . Couples\u2019 Extrinsic Emotion Regulation Questionnaire: Psychometric Validation in a Chilean Population. PLoS ONE2. 16(6): e0252329. https://doi.org/10.1371/journal.pone.0252329The original article (Kinkead & Riquelme,"} +{"text": "J Clin Invest. 2022;132(8):e149160. https://doi.org/10.1172/JCI149160Original citation: J Clin Invest. 2023;133(4):e169139. https://doi.org/10.1172/JCI169139Citation for this corrigendum: During the preparation of the manuscript, a histogram for The authors regret the error."} +{"text": "Journal of Biomedical Researchhttps://doi.org/10.7555/JBR.27.20120114,published on 30 September 2013.Correction to:We apologize for the misused images in"} +{"text": "Nature Communications 10.1038/s41467-022-34269-7, published online 15 November 2022Correction to: In this article the author name M. W. Krone was incorrectly written as W. M. Krone. The original article has been corrected."} +{"text": "Correction: Plant Methods (2021) 17:103 https://doi.org/10.1186/s13007-021-00802-wIn the Funding information section, the Grant Number was incorrectly given as \u2018No. 2019FYD1000900\u2019 and should have read \u2018No. 2019YFD1000900\u2019. The original article ["} +{"text": "Adv. Sci. 2019, 6, 190128010.1002/advs.201901280DOI: In the originally published article there is an error in Figure"} +{"text": "This article has been corrected: In 11964-11976. https://doi.org/10.18632/oncotarget.22600Original article: Oncotarget. 2018; 9:11964\u201311976."} +{"text": "There is an error in reference 11. The correct reference is:Tu K, Lu K, Zhang Q, Huang W, Xie D. Accurate single-cell genotyping utilizing information from the local genome territory. Nucleic Acids Res. 2021 Jun 4;49(10):e57. doi: 10.1093/nar/gkab106. PMID: 33619552."} +{"text": "Staphylococcus aureus are presented. They are closely related to prophages that were previously sequenced as part of S. aureus genomes.The annotated whole-genome sequences of five cultured phietaviruses infecting Staphylococcus aureus is a human commensal bacterium that has the potential to cause life-threatening infection (http://cab.spbu.ru/software/spades) (http://gravity.cvr.gla.ac.uk) related to SAP26 (GenBank accession number GU477322 [arbitrarily linearized]). The genomes were reoriented to reflect the termini of Staphylococcus prophages from a closely related genus . Genome annotation was performed as described previously in Galaxy (https://phrogs.lmge.uca.fr) (http://www.sbg.bio.ic.ac.uk/~phyre2/html/page.cgi?id=index) (http://trna.ucsc.edu/tRNAscan-SE) (http://rssf.i2bc.paris-saclay.fr/toolbox/arnold) (https://rfam.xfam.org/search#tabview=tab1) (http://genome2d.molgenrug.nl/g2d_pepper_promoters.php) (Paired-end 2 \u00d7 150-bp) sequencing using the Illumina DNA library preparation kit was performed on the NextSeq 2000 system at the Microbial Genome Sequencing Center (MiGS), which provided quality-controlled and trimmed reads. These reads were analyzed using the CPT Galaxy Phage Genome Assembler v2021.01 Workflow (axy-pub) with SPA/spades) , which pa.ac.uk) , which seviously , 11; open Galaxy and furt.uca.fr) and Phyrd=index) , and non0-bp sequ/arnold) , noncodiew=tab1) , and proers.php) , were iders.php) and SAMtS. aureus genomes .The five SAP genomes are ~43 kb , and porPRJNA857681) (Genomes are available in GenBank (see A857681) . The pha"} +{"text": "Nature Communications 10.1038/s41467-022-35384-1, published online 13 December 2022Correction to: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE196673 instead of https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE193673. This has been corrected in the PDF and HTML version of the article.The original version of this article contained an error in the \u201cData availability\u201d Statement. The hyperlink provided for the accession number GSE193673 was incorrectly given as"} +{"text": "This article has been corrected: In 58394-58404. https://doi.org/10.18632/oncotarget.16880Original article: Oncotarget. 2017; 8:58394\u201358404."} +{"text": "NORTH AMERICA28\u201031 July 2022American Podiatric Medical Association (APMA) Annual Scientific MeetingOrlando, Florida, USAwww.apma.org/events/thenational.cfm17\u201020 August 2022Annual Amputation Prevention Symposium (AMP)Chicago, Illinois, USAwww.amptheclimeeting.com/amp\u2010202226\u201028 August 2022Modern Wound Care ManagementStevenson, Wyoming USAmodernwound.comwww. 29 September \u2013 1 October 2022SVM 33rd Annual Scientific SessionsGrand Hyatt Denver, Denver, Colorado USAwww.woundsource.com/resource/svm\u201033rd\u2010annual\u2010scientific\u2010sessionsEUROPE19\u201020 July 2022International Conference on Novel Biomaterials in Drug Delivery and Wound Care ICNBDDWCCopenhagen, Denmarkwww.waset.org/novel-biomaterials-in-drug-delivery-and-wound-care-conference-in-july-2022-in-copenhagen15\u201018 August 2022International Surgical Week 2022 (ISW 2022)Vienna, Austriawww.isw2022.org7\u201010 September 2022International Continence Society Annual MeetingVienna, Austriawww.woundsource.com/resource/international\u2010continence\u2010society\u2010annual\u2010meeting7\u201310 September 202231st EADV CongressMilan, Italy & ONLINEwww.eadvcongress2022.org14\u201316 September 2022EPUAP 2022 Prague, Czech Republicwww.epuap2022.org14\u201317 September 2022Wounds Australia National Conference 2022Sydney, Australiawww.woundsaustralia.com.au/Web/Events/Web/Events/Event_List.aspx16\u201318 September 202218th Meeting of the Diabetic Foot Study Group (DFSG)Bratislava, Slovakiawww.dfsg.org/1/dfsg\u20102022"} +{"text": "Correction: International Archives of Occupational and Environmental Health 10.1007/s00420-022-01922-zhttps://legislation.wa.gov.au/.Work Health and Safety (Mines) Regulations 2022, Available online: https://www.business.qld.gov.au/industries/mining-energy-water/resources/safety-health/mining/hazards/dust/exposure-limits.Exposure limits for dust in mineral mines, Available online: Unfortunately the reference was wrongly published. The correct reference is given below."} +{"text": "Scientific Data10:89; 10.1038/s41597-023-01983-w, published online 11 February 2023.Correction to: In this article the author name Matthew R.V. Ross was incorrectly written as Matthew R. Ross. The original article has been corrected."} +{"text": "This article has been corrected: In 101224-101243. https://doi.org/10.18632/oncotarget.20642Original article: Oncotarget. 2017; 8:101224\u2013101243."} +{"text": "J Clin Invest. 2019;129(8):3214\u20133223. https://doi.org/10.1172/JCI125915Original citation: J Clin Invest. 2023;133(4):169317. https://doi.org/10.1172/JCI169317Citation for this corrigendum: \u2013/\u2013Pparb Opg-Fc sample. The correct figure is shown below.The authors recently became aware that an incorrect image was shown in The authors regret the error."} +{"text": "This article has been corrected: Due to errors during figure assembly, an incorrect blot was used for the p53 panel in 8270-8283. https://doi.org/10.18632/oncotarget.2013Original article: Oncotarget. 2014; 5:8270\u20138283."} +{"text": "Correction: Implement Sci 17, 42 (2022)https://doi.org/10.1186/s13012-022-01211-wFollowing publication of the original article , it was Additional file 1: Appendix A. School Professional Assessment Survey. Appendix B. School Professional Characteristics and Background. Appendix C. Re-Analysis Focusing on CBT Delivery Trends. Appendix D. Missing Data and Imputation."} +{"text": "Correction: BMC Neurol 22, 412 (2022)https://doi.org/10.1186/s12883-022-02926-5Following publication of the original article , the autAli M. AlamJian P. K. ChenGreta K. WoodBethany FacerManeesh BhojakKumar DasSylviane DefresAnthony MarsonJulia GranerodDavid BrownRhys H. ThomasSimon S. KellerTom SolomonBenedict D. MichaelThe original article has been"} +{"text": "J Clin Invest. 2022;132(11):e157549. https://doi.org/10.1172/JCI157549Original citation: J Clin Invest. 2023;133(2):e167843. https://doi.org/10.1172/JCI167843Citation for this corrigendum: In The authors regret the error."} +{"text": "This article has an addendum: The patient provided written informed consent.2137-2140. https://doi.org/10.18632/oncotarget.27598Original article: Oncotarget. 2020; 11:2137\u20132140."} +{"text": "J Clin Invest. 2022;132(8):e152187. https://doi.org/10.1172/JCI152187Original citation: J Clin Invest. 2022;132(10):e161559. https://doi.org/10.1172/JCI161559Citation for this corrigendum: Following the publication of this article, the authors became aware that an incorrect panel was used for The authors regret the error."} +{"text": "Consent was added to this article: The study, protocol # HRI-0029 is an IRB approved clinical study, WIRB # 20182804, and all patients signed informed consent to participate.1836-1847. https://doi.org/10.18632/oncotarget.28046Original article: Oncotarget. 2021; 12:1836\u20131847."} +{"text": "This article has been corrected: In 30992-31002. https://doi.org/10.18632/oncotarget.16047Original article: Oncotarget. 2017; 8:30992\u201331002."} +{"text": "Nature Communications 10.1038/s41467-022-29714-6, published online 19 April 2022.Correction to: In this article the author name Aurel B. Leuchtmann was incorrectly written as Aurel B. Leuchtman. The original article has been corrected."} +{"text": "This article has an addendum: The authors obtained written informed consent from the patient to publish information and images.1946-1952. https://doi.org/10.18632/oncotarget.28062Original article: Oncotarget. 2021; 12:1946\u20131952."} +{"text": "Catula gettyi gen. et sp. nov. (Lauraceae) from the Kaiparowits Formation . PLOS ONE 17(1): e0261397. https://doi.org/10.1371/journal.pone.0261397.The fourth author\u2019s name is spelled incorrectly. The correct name is: Joseph J.W. Sertich. The correct citation is: Maccracken SA, Miller IM, Johnson KR, Sertich JJW, Labandeira CC (2022) Insect herbivory on"} +{"text": "Consent was added to this article: All patients signed informed consent.4457-4462. https://doi.org/10.18632/oncotarget.27807Original article: Oncotarget. 2020; 11:4457\u20134462."} +{"text": "Scientific Reports 10.1038/s41598-022-27095-w, published online 27 December 2022Correction to: https://www.microsof.com/zh-cn/\u201d instead of \u201chttps://www.microsoft.com/zh-cn/\u201d.The original version of this Article contained an error in the legends of Figures 1, 4, 7 and 8, where there provided link was incorrectly given as \u201cAdditionally, the provided link in the legend of Figure 3 was incorrect. The legend,https://support.esri.com/en/Products/Desktop/arcgis-desktop/arcmap\u201d\u201cTypes of land use change trajectory. Software: PowerPoint 2021. URL: now reads:https://www.microsoft.com/zh-cn/.\u201d\u201cTypes of land use change trajectory. Software: PowerPoint 2021. URL: Lastly, this Article contained an error in the legend of Figure\u00a012, where the provided date was incorrect.https://support.esri.com/en/Products/Desktop/arcgis-desktop/arcmap.\u201d\u201cThe Moran scatter and LISA cluster graph of CSDR in Xinjiang from 1990 to 2015. Map generated with ArcGIS 10.8 (ESRI). URL: now reads:https://support.esri.com/en/Products/Desktop/arcgis-desktop/arcmap.\u201d\u201cThe Moran scatter and LISA cluster graph of CSDR in Xinjiang from 1990 to 2020. Map generated with ArcGIS 10.8 (ESRI). URL: The original Article has been corrected."} +{"text": "Correction: BMC Genomics 23, 709 (2022)https://doi.org/10.1186/s12864-022-08930-wFollowing publication of the original article , the autTherefore, the section should read as follows:Availability of data and materialshttps://www.ebi.ac.uk/ena/browser/view/PRJEB40421). fastq files for 21 domestic cat individuals can be found under ENA project PRJNA343389 (https://www.ebi.ac.uk/ena/browser/view/PRJNA343389).Newly generated whole genome individual sequencing data is available at European Nucleotide Archive (ENA) project number: PRJEB40421 (The original article has been"} +{"text": "J Clin Invest. 2017;127(1):169\u2013182. https://doi.org/10.1172/JCI89429Original citation: J Clin Invest. 2023;133(2):e168068. https://doi.org/10.1172/JCI168068Citation for this corrigendum: The authors recently became aware that representative illustrations presented in The updated"} +{"text": "This article has been corrected: In 86648-86659. https://doi.org/10.18632/oncotarget.13429Original article: Oncotarget. 2016; 7:86648\u201386659."} +{"text": "Diaporthe ilicicola is a newly described fungal species that is associated with latent fruit rot in deciduous holly. This announcement provides a whole-genome assembly and annotation for this plant pathogen, which will inform research on its parasitism and identification of gene clusters involved in the production of bioactive metabolites. Diaporthe species are ascomycete fungi found as saprophytes, endophytes, and plant pathogens . Data will inform research to understand how plant-pathogenic Diaporthe species differ from those that do not cause disease and will aid in the identification of fungal gene clusters involved in bioactive metabolite production.athogens and are athogens . This reDiaporthe ilicicola holotype strain FPH2015-502 (CBS 144318) was isolated as described by Lin et al. (Diaporthe ilicicola FPH2015-502 was grown in potato dextrose broth for 2\u2009days at 28\u00b0C and 150\u2009rpm. DNA was extracted with the DNeasy Plant minikit (Qiagen) according to the manufacturer\u2019s recommendations and sequenced with both short- and long-read sequencing. Illumina library preparation and sequencing were conducted at Novogene, Inc., using a NovaSeq 6000 system with paired-end 150-bp reads. The long-read sequencing library was prepared using the Oxford Nanopore Technologies SQK-LSK108 kit under standard preparation conditions with g-TUBE fragmentation. Long reads were sequenced using a Nanopore MinION Mk1B flow cell with R9.4.1 chemistry.2 CBS 14418 was isN50,\u20097,501\u2009bp) (Diaporthe helianthi strain 7/96 (GenBank accession number MAVT00000000) (D. helianthi 7/96 strain (GenBank accession number MAVT00000000) (Cytospora leucostoma (BioSample accession number SAMN04099706), Valsa mali (BioSample accession number SAMN03203459), Valsa malicola (BioSample accession number SAMN04099704), Valsa mali var. pyri (BioSample accession number SAMN03203462), and Valsa sordida (BioSample accession number SAMN04099705). The annotation was finalized using a modified version of OrthoFiller v1.2xonq (https://gitlab.com/xonq/orthofiller) referencing 15 Diaporthales species, namely, Coniella lustricola (https://mycocosm.jgi.doe.gov/Pilidi1/Pilidi1.home.html), Cryphonectria parasitica (BioSample accession number SAMN02744051), Cryptodiaporthe sp. (https://mycocosm.jgi.doe.gov/Crypto1/Crypto1.home.html), Cryptodiaporthe populea (https://mycocosm.jgi.doe.gov/Crypo1/Crypo1.home.html), Cytospora chrysosperma (https://mycocosm.jgi.doe.gov/Cytch1/Cytch1.home.html), Cytospora leucostoma (BioSample accession number SAMN04099706), Diaporthe ampelina (https://mycocosm.jgi.doe.gov/Diaam1/Diaam1.home.html) (Diaporthe citri (BioSample accession number SAMN15772025), Diaporthe helianthi (https://mycocosm.jgi.doe.gov/Diahe1/Diahe1.home.html) (Diaporthaceae sp. (https://mycocosm.jgi.doe.gov/DiaPMI573_1/DiaPMI573_1.home.html), Lollipopaia minuta (https://mycocosm.jgi.doe.gov/Lolmi1/Lolmi1.home.html), Melanconium sp. (https://mycocosm.jgi.doe.gov/Melsp1/Melsp1.home.html), Valsa mali , and Valsa sordida (BioSample accession number SAMN04099705).For genome assembly and annotation, default parameters were used for all software unless specified otherwise. Illumina read quality was checked using FastQC v0.11.8, and reads were trimmed using Trimmomatic v0.36 (options: HEADCROP:10 CROP:145 SLIDINGWINDOW:50:25 MINLEN:100) . Hybrid 0000000) with the0000000) , GeneMar0000000) , BUSCO v0000000) , and AUG0000000) . Protein0000000) , D. heliv1.2xonq (https:/me.html) , DiaportJALPVH000000000 .The genome was deposited in DDBJ/ENA/GenBank under the accession number"} +{"text": "This article has been corrected: In 1920-1936. https://doi.org/10.18632/oncotarget.28068Original article: Oncotarget. 2021; 12:1920\u20131936."} +{"text": "Open. Biol.11, 210199. (Published online 1 September 2021). (https://doi.org/10.1098/rsob.210199)Myoviridae in The"} +{"text": "This article has been corrected: In 2141-2159. https://doi.org/10.18632/oncotarget.27329Original article: Oncotarget. 2020; 11:2141\u20132159."} +{"text": "First published: April 13, 2022https://onlinelibrary.wiley.com/doi/10.1111/jvim.16412Volume 36, Issue 3Correction to authorship information as follows.The Journal of Veterinary Internal Medicine does not allow 2 corresponding authors or 2 first authors. Dr. Anna R. Gelzer is the corresponding author, not Dr. Cory M. Taschabrunn. Drs. Alexandra V. Crooks and Weihow Hsue contributed equally in their author roles."} +{"text": "J Clin Invest. 2022;132(19):1\u201318. https://doi.org/10.1172/JCI157399Original citation: J Clin Invest. 2023;133(3):e168771. https://doi.org/10.1172/JCI168771Citation for this corrigendum: After the publication of this article, the authors became aware that the \u03b2-actin blot shown in The authors regret the error."} +{"text": "This article has been corrected:3681-3693. https://doi.org/10.18632/oncotarget.26978Original article: Oncotarget. 2019; 10:3681\u20133693."} +{"text": "This article has been corrected: In 26527-26542. https://doi.org/10.18632/oncotarget.25480Original article: Oncotarget. 2018; 9:26527\u201326542."} +{"text": "J Clin Invest. 2023;133(1):e153733. https://doi.org/10.1172/JCI153733Original citation: J Clin Invest. 2023;133(4):e169299. https://doi.org/10.1172/JCI169299Citation for this corrigendum: The title of this paper was incorrect. The correct title is above. The HTML and PDF versions have been updated online. The authors regret the error."} +{"text": "Phil. Trans. R. Soc. B377, 20210178. (Published 21 September 2022) (https://doi.org/10.1098/rstb.2021.0178)One of the authors' names was spelt incorrectly as Addsion Billing instead of Addison Billing.This has now been corrected on the publisher's website."} +{"text": "Correction: Inflamm Regen 42, 35 (2022)https://doi.org/10.1186/s41232-022-00221-xFollowing publication of the original article , the authttps://www.genome.jp/kegg/. Accessed 27 Jul 2022.48. Kyoto Encyclopedia of Genes and Genomes. The correct version is:48. Kanehisa M, Goto S. KEGG: Kyoto Encyclopedia of Genes and Genomes. Nucleic Acids Res. 2000;28:27\u201330.The original article has been"} +{"text": "This article has been corrected: In 19172-19191. https://doi.org/10.18632/oncotarget.13432Original article: Oncotarget. 2017; 8:19172\u201319191."} +{"text": "Community Eye Health Journal will be on Neuro-ophthalmology. It will include articles such as \u2018Understanding vision and the brain\u2019 and \u2018Assessing the neuro-ophthalmology patient\u2019. This issue will not be produced in paper format because of increasing costs in publication and distribution. It will be available online at www.cehjournal.com If you wish to receive an email with a link to download the PDF copy, please send your email address to web@cehjournal.org.The next issue of the Continued Professional Development. Thank you for your understanding.The next paper issue, planned for the end of March 2017, will be on IAPB's General Assembly (10GA), the premier global event discussing public health issues related to blindness and visual impairment, brought together 1,150 eye care professionals from 100 countries in Durban in October 2016.Over the course of three days, there were over 60 sessions with 200 speakers, and over 250 poster presentations. If you couldn't be there, you can catch up on what you've missed:www.iapb.org/10ga-presentationsView and download PowerPoint files of all the talks and presentations from http://atlas.iapb.orgAccess IAPB's Vision Atlas, which was launched at the 10GA. It allows access to the latest data and evidence related to avoidable blindness and sight loss: http://photocomp.iapb.orgEnjoy the photographs entered into the #StrongerTogether Photo Competition. The winners were announced at 10GA and all entries can be viewed at vtc@gju.edu.joEmail: www.health.uct.ac.za or email chervon.vanderross@uct.ac.zaWrite to the Training Coordinator, Lions Medical Training Centre, Lions SightFirst Eye Hospital, PO Box 66576-00800, Nairobi, Kenya.Tel: +254 20 418 32 39www.kcco.net or contact Genes Mng'anga at genes@kcco.netVisit www.cehjournal.org/subscribeadmin@cehjournal.orgFor paper copies, email Anita Shah: web@cehjournal.orgTo receive an alert when a new issue is published, email www.cehjournal.orgVisit us online: www.facebook.com/CEHJournal/https://twitter.com/CEHJournal Lindsley K, Matsumura S, Hatef E, Akpek EK. Interventions for chronic blepharitis. Cochrane Database Syst Rev. 2012, 5: CD005556. doi: 10.1002/14651858. CD005556.pub2Clearfield E, Muthappan V, Wang X, Kuo IC. Conjunctival autograft for pterygium. Cochrane Database Syst Rev. 2016, 2: CD011349. doi: 10.1002/14651858. CD011349.pub2.Ervin AM, Wojciechowski R, Schein O. Punctal occlusion for dry eye syndrome. Cochrane Database Syst Rev. 2010, 9: CD006775. doi: 10.1002/14651858. CD006775.pub2.Wilhelmus KR. Antiviral treatment and other therapeutic interventions for herpes simplex virus epithelial keratitis. Cochrane Database Syst Rev. 2015, 1:CD002898. doi: 10.1002/14651858.CD002898.pub5.Gichuhi S, Irlam JH. Interventions for squamous cell carcinoma of the conjunctiva in HIV-infected individuals. Cochrane Database Syst Rev. 2013 28; 2:CD005643. doi: 10.1002/14651858. CD005643.pub3.Gipson IK. The ocular surface: the challenge to enable and protect vision: the Friedenwald lecture. Invest Opht Vis Sci 2007;48(10):4390; 4391-4398.https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2886589/Available online: www.tearfilm.org/dewsreport/Methodologies to diagnose and monitor dry eye disease: report of the Diagnostic Methodology Subcommittee of the International Dry Eye Workshop (2007). Ocul Surf. 2007;5(2):108-52. Available online: http://tinyurl.com/dry-eye-circleBaudouin C, Messmer EM, et al. Revisiting the vicious circle of dry eye disease: a focus on the pathophysiology of meibomian gland dysfunction. BJO Online First, published on January 18, 2016. Available online:"} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-022-26760-4, published online 05 January 2023Correction to: The original version of this Article contained an error in Reference 37, which was incorrectly given as:et al. Monogalactosyldiacylglycerol deficiency in tobacco inhibits the cytochrome b6f-mediated intersystem electron transport process and affects the photostability of the photosystem II apparatus. Biochim Biophys. Acta709\u2013722, 2013. https://doi.org/10.1016/j.bbabio.2013.02.013\u00a0(1827).Wu, W. The correct reference is listed below:et al. Monogalactosyldiacylglycerol deficiency in tobacco inhibits the cytochrome b6f-mediated intersystem electron transport process and affects the photostability of the photosystem II apparatus. Biochim. Biophys. Acta1827, 709\u2013722. https://doi.org/10.1016/j.bbabio.2013.02.013 (2013).Wu, W. The original Article has been corrected."} +{"text": "Correction to: BMC Bioinformatics (2022) 23:448https://doi.org/10.1186/s12859-022-04983-6Following publication of the original article , the autThe original article has beenAdditional file 1. supp file 1: filt3r-supp.pdf.Additional file 2. supp file 2: results_sra.xlsx.Additional file 3. supp file 3: accessions.csv.Additional file 4. supp file 4: results-different-k.xlsx.Additional file 5. supp file 5: results-small-k.xlsx.Additional file 6. supp file 6: results-different-perc.xlsx.Additional file 7. supp file 7: results-subsamples.xlsx.Additional file 8. supp file 8: genescan_SRR15006459.pdf.Additional file 9. supp file 9: genescan_SRR15006372.pdf. Additional file 10. supp file 10: accessions-CCLE.csv.Additional file 11. supp file 11: results_simulated_150bp.xlsx.Additional file 12. supp file 12: results_simulated_250bp_hq.xlsx. Additional file 13. supp file 13: results_simulated_250bp_lc.xlsx.Additional file 14. supp file 14: results_simulated_250bp_lq.xlsx.Additional file 15. supp file 15: results_simulated_250bp.xls."} +{"text": "This article has been corrected: In 23020-23032. https://doi.org/10.18632/oncotarget.15479Original article: Oncotarget. 2017; 8:23020\u201323032."} +{"text": "In the published article, there was an error with a reference as follows:Exp Toxicol Pathol. (2007) 58:367\u201374. doi: 10.1016/j.etp.2006.11.006.21. Eoi: 10.1007/BF02686024dy of the effects of cadmium and copper on fefails to reduce cadmium-induced oxidative damage in rat liver. The correct reference is below:Exp Toxicol Pathol. (2007) 58:367\u201374. doi: 10.1016/j.etp.2006.11.006.21. E\u015frefoglu M, G\u00fcl M, Dogru MI, Dogru A, Y\u00fcrekli M. Adrenomedullin fails to reduce cadmium-induced oxidative damage in rat liver. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "This article has an addendum: All human tissues were sourced from the IUCPQ Biobank, which includes only patients that have signed a consent form for research.209-220. https://doi.org/10.18632/oncotarget.27874Original article: Oncotarget. 2021; 12:209\u2013220."} +{"text": "This article has been corrected: In 1894-1910. https://doi.org/10.18632/oncotarget.27585Original article: Oncotarget. 2020; 11:1894\u20131910."} +{"text": "This article has been corrected:183-197. https://doi.org/10.18632/oncotarget.28179Original article: Oncotarget. 2022; 13:183\u2013197."} +{"text": "Psychoradiology were mistakenly published into Precision Clinical Medicine Volume 1, Issue 1, June 2018.In April 2022, three articles intended for the journal The articles were:https://doi.org/10.1093/psyrad/kkaa001https://doi.org/10.1093/psyrad/kkab001https://doi.org/10.1093/psyrad/kkaa002Psychoradiology, Volume 1, Issue 1, March 2021. The publisher apologizes for this error.These articles have now been moved to"} +{"text": "Correction to: BMC Women's Health (2022) 22:427 10.1186/s12905-022-02004-5Following publication of the original article , the aut66. World Medical Association. World Medical Association Declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA. 2013;310(20):2191\u20132194. 10.1001/jama.2013.281053.Also, the in text citation has been changed in the original article.The original article has been corrected."} +{"text": "This article has an addendum: Informed patient consent was obtained.4836-4844. https://doi.org/10.18632/oncotarget.27848Original article: Oncotarget. 2020; 11:4836\u20134844."} +{"text": "This article has an addendum: Patient consent was obtained from the patient. This study was approved by the ethical review board of Centre L\u00e9on-B\u00e9rard.1618-1628. https://doi.org/10.18632/oncotarget.27563Original article: Oncotarget. 2020; 11:1618\u20131628."} +{"text": "JCI Insight. 2022;7(13):e136678. https://doi.org/10.1172/jci.insight.136678Original citation: JCI Insight. 2022;7(15):e163757. https://doi.org/10.1172/jci.insight.163757Citation for this erratum: During the preparation of this manuscript, JCI regrets the error.The"} +{"text": "Phil. Trans. R. Soc. B376, 20200086. (Published 17 May 2021). (doi:10.1098/rstb.2020.0086)Some funding information was inadvertently omitted from the funding statement.This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Sk\u0142odowska-Curie grant agreement no. 841127."} +{"text": "This article has been corrected: In 131-147. https://doi.org/10.18632/oncotarget.27413Original article: Oncotarget. 2020; 11:131\u2013147."} +{"text": "First published: 10 June 2021https://onlinelibrary.wiley.com/doi/10.1111/jvim.16197Volume 35, Issue 4Correction to the grant/award number for the funder as follows.Funding information: Rural Development Administration, Grant/Award Number: PJ01404502."} +{"text": "J Clin Invest. 2018;128(2):861\u2013875. https://doi.org/10.1172/JCI96218Original citation: J Clin Invest. 2022;132(16):e163716. https://doi.org/10.1172/JCI163716Citation for this corrigendum: The authors recently became aware of errors in The authors regret the errors."} +{"text": "Senior editors, editors, reviewers, authors, and the American Society for Microbiology (ASM) staff all play important, complementary roles in the scientific publishing process. In 2022, we found ourselves in somewhat uncertain times as the ASM Journals program continued its transition to fully open access. Nonetheless, we are confident that mSphere is well positioned, having been open access since our inception, and with our impact continuing to grow each year. We are encouraged that authors have also voted with their submit buttons, so to speak, continuing to publish their excellent work with us.We have a great end-of-year tradition at Raquel AbadAzar AbadiCelina Monteiro AbreuMichael C. AbtDenise M. AkobKathirvel AlagesanBassem AllamRey Custer AllenRichard A. AlmChristopher AlteriJohn AlverdyAlfred Amambua-NgwaFrancisco AmaroMatthew Zack AndersonLaura Maria Andrade de OliveiraAlex AndrianopoulosLinda S. ArchambaultRobert A. ArkowitzChelsie Elizabeth ArmbrusterPaul M. ArnaboldiGustavo ArrizabalagaSassan AsgariNick AshboltJennifer M. AuchtungMatthew B. AvisonSteffen BackertAnjana BadrinarayananJuhi BagaitkarLauren O. BakaletzJonathon L. BakerScott BalibanJimmy D. BallardMarcela Passos Galluzzi BaltazarJonathan M. BaraschRoman Alfredo BarcoBrianne BarkerSamuel BarnettJavad BaroueiJeremy J. BarrStephen D. BarrLuther A. BarteltChristine Marie BassisAndrea BattistoniClifford J. BeallJessica A. BelserBenjamin Ross BelvinMar BenavidesMaureen BergMegan BergkesselDebra E. BessenKyle BibbyKevin BickerPablo BifaniBlake BillmyreCraig BingmanBrian BirdLogan BlancettJill R. BlankenshipJoseph M. BlissPatricia Pringle BloomKarl W. BoehmeLydia M. BogomolnayaJon BohlinM\u00e9lanie BonhiversSelina BoppCristian BottaMichael John BotteryTeun BousemaKate BowermanAndrew S. BowmanAllison BradyJonathan BraunMichela BrazzoliBenjamin BrennanDavid J. BrennerChristopher B. BrookeJeremy C. BrownlieEmily A. BruceHarry BrumerCarmen BuchrieserJerry M. BuysseMatthew T. CabeenEdward M. CampbellShengbo CaoGeorge CarnellBerit CarowJaime CarrascoJuan Manuel CarrenoMark CarringtonSarah R. CarterFred CasselsMarco CassoneRoberto CattaneoRodrigo Cay\u00f4Miguel Angel CevallosBenjamin J. ChadwickChrispin ChaguzaGeorgios ChamilosClara S. ChanJeffrey ChandlerYung-Fu ChangMuhammad Tausif ChaudhryHuirong ChenLiang ChenYe ChenHae Suk CheongMichaelle ChojnackiRebecca C. ChristoffersonAlexander T. CiotaStephen A. ClarkDavid J. ClarkeChristine ClaytonBryan CobbShira Milo CochaviDarrell CockburnJames CollinsSean ConlanBede ConstantinidesLaura CookLaura C. CookIsabelle CoppensTeresa CoqueAngelina CordoneCynthia Nau CornelissenJohn R. CortDoug CossarGeorgina CoxRebecca Jane CoxKathryn CoyneRobert A. CramerNeil CrickmoreAlan CrossLiwang CuiPaul J. CullenKathleen CusickMichael CynamonWei DaiStephen DanielsHeran DarwinSiddhartha DasChandravanu DashRaymond James DattwylerMary E. DaveyMark R. DaviesApril Dawn DavisZeger DebyserMaxime DeforetElizabeth De GaspariChristopher Luis De GraffenriedYe DengNicole J. De NiscoJigar V. DesaiGirmay DesalegnSantosh DhakalVijaykrishna DhanasekaranThomas DickAntonio DiGiandomenicoFrancisco DionisioRichard D. DixYohei DoiMichael S. DonnenbergCurtis DonskeyCharles J. DormanGoncalo dos Santos CorreiaZhicheng DouDavid J. DowlingTimothy B. DoyleJuan DuEdward G. DudleyBreck A. DuerkopJoseph Alexander DuncanJulie C. Dunning HotoppFrank EbelLeo EberlAdrianne N. EdwardsAndrew EdwardsMelissa EllermannJeremy R. EllermeierMostafa S. ElshahedIuliana V. EneOthmar G. EngelhardtMelinda Anne EngevikMarkus EngstlerEeva Liisa Eronen-RasimusJavier Antonio Escobar-PerezJos\u00e9 A. EscuderoVanessa EspinosaMorgan EvansChristina S. FahertyRefath FarzanaYingang FengYoujun FengKenneth A. FieldsD. Clark FilesMelanie J. FiliatraultLaura FilkinsStefan FinkeDerek J. FisherFabrizio FoieniEdward M. FoxMichael T. FranceBettina C. FriesFriedrich FrischknechtErnesto J. FuentesIwona GabrielStarrett GabrielMichaela Ulrike GackAttila GacserJennifer GaddyRaj GajiEmily GallichotteKetaki GantiDar\u00edo Garc\u00eda De ViedmaMeritxell Garcia-QuintanillaGregory GavelisMakda GebreJennifer Geddes-McalisterLee GehrkeLorenzo GiacaniTim GilbergerSteven R. GillJoseph James GillespieSusan S. GoldenGustavo H. GoldmanVijay GondilTobias GorisThomas GorochowskiRia GoswamiRevathi GovindLone GramTimothy J. GreenElisabeth GrohmannTrudy H. GrossmanSebastian GuentherPascale GuitonArthur G\u00fcnzlShashank GuptaEyal GurKiran GurungYitzhak HadarMaria HadjifrangiskouAndrea HahnMohamed A. HakimiRoy A. HallParis S. HammJun HangGeoffrey D. HanniganKyle HappelMd. Manjurul HaqueJustin HardickChristian M. HardingReuben S. HarrisBen M. HauseCynthia Y. HeJianzhong HeNicholas S. HeatonTory A. HendrySteven HennigarFrancisca Hernandez-HernandezLena J. HeungMichael HolickPeiying HongAlexander R. HorswillDaniel K. HoweFupin HuJulian G. HurdleJillian H. HurstBonnie L. HurwitzKevin HybiskeAshraf IbrahimKaoru IkumaIliyan IlievRalph R. IsbergHideomi ItohAngela IvaskGulnaz T. JavanVicki JeffersKelsea A. JewellZhilong JiaBaoming JiangCheng JinJames R. JohnsonWilliam JohnsonClinton J. JonesSheryl JusticeBj\u00f6rn F. C. KafsackBarbara C. KahlJeremy Phillip KamilJames B. KaperThomas E. Kehl-FieBrendan KellyVolkhard A. J. KempfLily KhadempourM. Firoze KhanShahid M. KhanAnagha KhandekarMegan R. KiedrowskiNicolas KiefferHye Kwon KimPeter E. KimaJohn H. KimbroughSamantha Jane KingStephen KisslerTomoe KitaoEllen KneuferShintaro KobayashiTheresa M. KoehlerMichael H. KogutJames B. KonopkaAnna KonovalovaNicole Marie KoropatkinIoly Kotta-LoizouLukasz KozubowskiFlorian KrammerBarry N. KreiswirthInna V. KriegerErik KristianssonJames W. KronstadCarol A. KumamotoAnand KumarAnuj KumarMukesh KumarAshlan J. Kunz CoyneKyohei KurodaHiroyuki KusadaManish KushwahaJoseph KusiOlaf KutschMamuka KvaratskheliaAbigail L. LabellaBorden LacyMonique LafonGyanu LamichhaneTilman LamparterChung-Yu LanKristin LaneStephanie N. LangelDenis LeclercAllen LeeVincent T. LeeFrancois Le MauffJose M. Lemme DumitSebastian LeptihnCammie F. LesserMichael LetkoJiasui LiLing LiYong LiGeorge LiechtiEfrem S. LimJean LimChi-Hung LinYongxin LinDaniel LindnerNing LingVincenzo LionettiRosalia LiraBinbin LiuJia LiuOlga LomovskayaJason S. LongS. Wesley LongAnja L\u00fchrmannBrian Michael LunaLiang MaIain MacarthurAntonio MachadoMatthias P. MachnerMarisa MadridFinlay MaguireAndrea MaisnerMichael B. MajorChrister MalmbergAlexander MankinSam MannaShannon D. ManningNicholas J. MantisKevin MaringerClaudia N. H. MarquesFumito MaruyamaSeverine MatheusRosario MatoRobin C. MayJennifer A. MaynardJere W. McBrideShonna M. McBrideBeth A. McCormickElizabeth A. McDanielPatrick McGannLesley McGeeJoy A. McKennaJoseph B. McPheeWim G. MeijerJason MercerTimothy C. MeredithAlita A. MillerKathryn C. Milligan-McClellanCarmen MirabelliDominique MissiakasHeungyun MoonIain M. MorganJames C. MorrisKarl M\u00fcngerAlison E. MurrayMario E. MuscarellaEleftherios MylonakisJoe S. MymrykMiki NagaoRyosuke NakaiYu NakajimaJadranka NappiFernando Navarro-GarciaEsther NdungoDavid R. NelsonMartha I. NelsonMinh Hong NguyenWright W. NicholsHelge NiemannLeonardo NimrichterLaura Mary NolanSteven J. NorrisFernanda NovaisDennis NurjadiJoshua J. ObarAllyson F. O'DonnellFrank OechslinKazuhiro OgaiToru OkamotoYusuke OkazakiDaniel Groban OlsonCarlos J. OrihuelaEva Ortega-RetuertaKenji OtaHong-Yu OuBrian PalenikGlen E. PalmerRobert J. PalmerJohn C. PanepintoHuili PangHee-Soo ParkKathryn A. PatrasSara F. PaverJ. S. Malik S. M. PeirisDaniel R. PerezVincent PerretenAndreas PeschelHadrien PeyretCatherine Ann PfisterJody PhelanGorben Peter PijlmanVittal Prakash PonrajMegan PovelonesAditi PrabhakarJacob PriceLance B. PriceBali PulendranDohun PyeonLeimin QianAlison J. QuayleMaxime Qu\u00e9batteRobert Andrew QuinnTara M. RandisTara M. RandisGauri G. RaoJayne RaperChad A. RappleyeJeremy RatcliffRaveen RathnasingheChristina R. RathwellMichael L. ReeseAaron ReinkeJustin RemaisNilton RennoPeter A. RiceDave RichardJuergen A. RichtJames RiddellDouglas RisserAmariliz RiveraDaniel D. RockeyG. Marcela RodriguezJose-Manuel Rodriguez-MartinezSandra Romero-SteinerMartin RottmanClaudia R\u00fcckertElizabeth Ann RucksYang RuiThomas A. RussoKathrin RychliRobert SabatiniJaiprasath SachithanandhamMarat R. SadykovXavier SaelensMohammad Mohseni SajadiDavinia Salvach\u00faa RodriguezDerrick SamuelsonJohn C. SamuelsonNicholas D. SandersonSarah E. SansomPanagiotis SapountzisLarry S. SchlesingerThomas M. SchmidtDirk SchnappingerMichael SchuitStefan SchwarzPhillip ScottKelly E. SeatonPatrick R. SecorAnna Maria SeekatzWilliam SelfAnna SelmeckiChetan SeshadriRebecca S. ShapiroAnupam SharmaRoss ShawLilach SheinerAimee ShenTrevor ShoemakerAnton M. SholukhErika ShorL. David SibleyJeffrey SiegelBaneshwar SinghCiaran SkerryJoseph SmithWiep Klaas SmitsSandro Gomes SoaresStephanie D. SongJoseph A. SorgShanmuga SozhamannanJennifer K. SpinlerStanley M. SpinolaShiranee SriskandanRichard StantonGabriel J. StarrettChad SteeleDavid S. StephensWolfgang R. StreitMikael Lenz StrubeXin-Zhuan SuKarthik SubramanianYo SugawaraDaigo SumiNobuhiro SuzukiYasuhiro SuzukiMary Hannah SwaneyJason B. SylvanClifford C. TaggartAkifumi Takaori-KondoMichal Caspi TalRita TamayoSatoshi TamazawaYing TaurSam R. TelfordLesly A. TemesvariSharon Mei TennantBenno Herman Ter KuileRita TewariElitza S. TheelKevin R. TheisTorsten ThomasCecilia ThompsonYun TianMaria TomasDieter M. TourlousseMasanori ToyofukuKatrina E. TraberJohn S. TregoningAndrea TrevisanDavid R. TribbleDavid TurraJane F. TurtonYuki UeharaSvetlana Ugarcina PerovicElizabeth R. UngerJane UsherFrancisco UzalKrystal VailChristiaan van OoijJan-Peter van PijkerenCaroline Elisabeth VisserRomain VolleJay VornhagenSarah VreugdeWillem WaegemanSeth T. WalkDavid H. WalkerDavid A. WalshShanquan WangChongzhen WangLuxin WangTian WangTiffany WeinkopffDavid S. WeissLouis M. WeissVolkmar WeissigStephen Robert WelchKristen E. WendtEric WenzlerEdze WestraDawn WetzelAnnegret WildeMichael R. WileyMary E. WilsonSteven S. WitkinKenneth H. WolfeManuel WoltersPatrick C. Y. WooR. Mark WootenFloyd L. Wormley, Jr.Karen L. WozniakRachel A. F. WozniakXianfu WuXin XuChaoyang XueTetsuo YamaguchiKyosuke YamamotoDong YangYiling YangOzlem YilmazHyunah YoonMikaeel YoungJae-Hyuk YuYunsong YuJoseph P. ZackularMar\u00eda Mercedes ZambranoMerve S. ZedenJin ZengBing ZhaiKai ZhangShengda ZhangWei ZhangWeiping ZhangZhaolei ZhangJiangchao ZhaoXian-Liang ZhaoXilin ZhaoChunfu ZhengHaixue ZhengQingfei ZhengXiaoxian ZhongPanpan ZhouErwin G. ZoetendalThe breadth of our scope, encompassing the microbial sciences writ large, means that we depend on a large and diverse pool of reviewers to guide our decisions about manuscripts. Those individuals who helped us this year are listed here and have our sincere gratitude."} +{"text": "An updated species checklist for all the Pentatomoidea species for Greece is provided. Eight species are recorded from Greece for the first time. The checklist is supported with distributional data notes for all the Pentatomoidea species of Greece.Aelia germari K\u00fcster 1852, Eurygaster hottentotta (Fabricius 1775), Eysarcoris aeneus (Scopoli 1763), Neottiglossa lineolata (Herrich-Schaeffer 1830), Neottiglossa pusilla , Picromerus bidens (Linnaeus 1758), Podops (Podops) inunctus (Fabricius 1775) and Tarisa pallescens (Jakovlev 1871). A complete updated species checklist with distributional data notes for all the new species for Greece are provided.Eight species of the superfamily Pentatomoidea are recorded from Greece for the first time: Halyomorpha halys (St\u00e5l) and the southern green stink bug Nezara viridula (L.).The superfamily Pentatomoidea constitutes one of the most important insect groups of the suborder Heteroptera. It includes 1080 genera and 5907 species belonging to 16 families of which the Cydnidae, Pentatomidae, Scutelleridae and Tessaratomidae are the most important; 94% of the species belong to these four families ,3. PentaHalyomorpha halys is a polyphagous stink bug native to China, Korea, Japan and Taiwan , Olympos Mt. [Prionia (21.V.1981)]. Total: 3 specimens.Elasmucha St\u00e5l 1864 (Genus t\u00e5l 1864 2.Elasmucha grisea grisea (Linnaeus 1758)Doris [Skaloula (2.V.1986)], Ioannina (24.VI.1984), Rodopi Mt. (10.VIII.1985). Total: 5 specimens.Family CYDNIDAESubfamily CydninaeCydnus Fabricius 1803 (Genus ius 1803 3.Cydnus aterrimus (Forster 1771)Santorini Island [Kamarion (5.V.1982)]. Total: 1 specimen.Macroscytus Fieber 1860 (Genus ber 1860 4.Macroscytus brunneus (Fabricius 1803)Attiki [Kifissia ], Doris [Agios Nikolaos (2.VII.1986)]. Total: 4 specimens.Subfamily SehirinaeOchetostethus Fieber 1860 Doris , Olympos Mt. . Total: 5 specimens.Tritomegas Amyot et Serville 1843 (Genus lle 1843 6.Tritomegas bicolor (Linnaeus 1758)Epirus [Vryssochori (28.V.1981)], Voeotia [Arachova (17.III.1981)]. Total: 2 specimens.7.Tritomegas sexmaculatus (Rambur 1839)Olympos Mt. [Stavros (20.V.1981)]. Total: 1 specimen.Family PENTATOMIDAESubfamily AsopinaeArma Hahn 1832 Genus 8.Arma insperata (Horv\u00e1th 1899)Florina [Vernon Mt-Kalo Nero (21.VIII.1986)]. Total: 1 specimen.Jalla Hahn 1832 Genus 9.Jalla dumosa (Linnaeus 1758)Doris [Skaloula (1.VI.1985)], Fokis [Elaion (22.VII.1983)], Kozani [Vourinos Mt. 18.VIII.1983)]. Total: 3 specimens.Picromerus Amyot et Serville 1843 Genus 10.Picromerus bidens* (Linnaeus 1758) (us 1758) Florina [Vernon Mt-Kalo Nero (30.VII.1982)]. Total: 3 specimens.11.Picromerus conformis (Herrich-Schaeffer 1841)Doris [Skaloula (27.IX.1983)], Florina [Psarades (18.VIII.1986)]. Total: 2 specimens.Troilus St\u00e5l 1867 (Genus t\u00e5l 1867 12.Troilus luridus (Fabricius 1775)Olympos Mt. [Stavros (20.V.1981)]. Total: 1 specimen.Zicrona Amyot et Serville 1843 (Genus lle 1843 13.Zicrona caerulea (Linnaeus 1758)Attiki [Avlon (11.V.1980)], Doris [Monastirakion (18.VIII.1981)]. Total: 2 specimens.Subfamily PentatominaeAcrosternum Fieber 1860 Genus 14.Acrosternum heegeri (Fieber 1861)Doris [Eratini (14.VII.1978)], Zakynthos [Volimes (28.VI.1988)], Athos [Karyes (5.VIII.1986)]. Total: 7 specimens.15.Acrosternum millierei (Mulsant et Rey 1866)Attiki [Marathon (11.VII.1984)], Chios [Chora (5.IV.1979)], Doris [Agios Nikolaos (20.VII.1986)], Kynouria [Astros (31.VIII.1984)]. Total: 9 specimens.Aelia Fabricius 1803 Genus 16.Aelia acuminata (Linnaeus 1758)Aetoloakarnania [Messolonghion (17.V.1990)], Amphilochia [Anixiatiko (20.VIII.1985)], Arkadia [Menalon Mt. (20.VI.1986)], Attiki , Chalkidiki [Valti (8.VIII.1985)], Chios [Chora ], Doris , Epirus [Vryssochori ], Evia [Oreoi (20.IV.1980)], Evrytania [Megalo Chorio (10.VIII.1986)], Drama [Falakron Mt. (13.VI.1982)], Florina , Fokis [Galaxidi (15.IV.1980)], Fthiotis , Grevena [Anoixis ], Ikaria , Ioannina , Karditsa [Artessiano (1.X.1981)], Konitsa [Aoos Riv. (29.V.1981)], Lakonia [Mystras (30.IV.1985)], Lesvos [Sykamnia (16.VI.1987)], Messinia , Naxos [Apeiranthos (16.VI.1981)], Olympos Mt. (13.VIII.1979), Parnassos Mt. [National Park (13.VII.1985)], Paros [Paroikia (18.VI.1981)], Pella [Vryta (14.VIII.1979)], Pieria [Poroi ], Pindos Mt. , Rodopi Mt. [Elatia ], Rhodos [Dimilia (31.V.1990)], Strymonas Riv. (1.VI.1982), Thessaloniki , Trikala [Orthovounion (30.IX.1981)], Vegoritis Lake (14.VIII.1979), Xanthi [Porto Lagos ], Zakynthos [Keri (29.VI.1988)]. Total: 119 specimens.17.Aelia albovittata (Fieber 1868)Chios [Armolia (20.VI.1987)], Samos [Pythagorion (23.VI.1987)]. Total: 3 specimens.18.Aelia germari* (K\u00fcster 1852) (er 1852) Evros [Metaxades (2.VI.1982)]. Total: 1 specimen.19.Aelia klugii (Hahn 1831)Florina [Vernon Mt. Kalo Nero ], Rodopi [Vathyrhemma (12.VI.1982)]. Total: 5 specimens.20.Aelia rostrata (Boheman 1852)Attiki , Doris [Ghiona ], Drama [Potami (12.VI.1982)], Epirus [Vryssochori (28.V.1981)], Evros , Florina ), Fthiotis [Anthili (12.VIII.1980)], Konitsa [Aoos Riv. (30.V.1981)], Kozani [Vourinos Mt. (18.VIII.1985)], Messinia , Oiti Mt. ], Olympos Mt. [Stavros (20.V.1981)], Parnon Mt. (18.VI.1986), Prionia (30.V.1982), Rhodos [Plymiri (29.VI.1987)], Rodopi Mt. [Elatia (19.VII.1983)]. Total: 38 specimens.21.Aelia virgata (Klug 1841)Evros [Metaxades (2.VI.1982)], Trikala [Aghiofilo (22.VII.1989)]. Total: 2 specimens.Antheminia Mulsant et Rey 1866 (Genus Rey 1866 22.Antheminia lunulata (Goeze 1778)Fthiotis [Kalamakion (12.VII.1979)], Pella [Vryta (14.VIII.1979)], Vegoritis Lake (14.VIII.1979). Total: 3 specimens.Apodiphus Saunders 1877 Attiki , Doris [Skaloula (5.X.1980)]. Total: 7 specimens.Brachynema Mulsant et Rey 1852 (Genus Rey 1852 24.Brachynema cinctum (Fabricius 1775)Attiki [Schinias (1.VII.1982)]. Total: 1 specimen.Carpocoris Kolenati 1846 (Genus ati 1846 25.Carpocoris (Carpocoris) fuscispinus (Boheman 1851)Attiki , Doris [Skaloula ], Epirus [Vryssochori (27.V.1981)], Evia , Kozani [Vourinos Mt. ], Parnon Mt. , Messinia [Kazarma (4.VII.1984)], Olympos Mt. [Prionia (13.VIII.1979)], Paros [Paroikia (18.VI.1981)], Rhodos , Samos , Oiti Mt. (9.VIII.1986), Thessaloniki [Cedron Hills (14.XI.1982)], Vegoritis Lake (14.VIII.1979). Total: 38 specimens.26.Carpocoris (Carpocoris) mediterraneus mediterraneus (Tamanini 1958)Evros Riv. [Delta (7.VI.1982)], Naxos [Moutsouna (15.III.1982)]. Total: 4 specimens.27.Carpocoris (Carpocoris) pudicus (Poda 1761)Attiki , Doris [Skaloula ], Florina [Vevi (24.VIII.1983)], Rodopi Mt. [Betoula Forest (22.V.1983)], Konitsa [Aoos Riv. (30.V.1981)], Kozani [Vourinos Mt. (17.VI.1985)], Vegoritis Lake (14.VIII.1979). Total: 10 specimens.Chlorochroa St\u00e5l 1872 (Genus t\u00e5l 1872 28.Chlorochroa sp. St\u00e5l 1872Olympos Mt. [Stavros (20.V.1981)]. Total: 1 specimen.Codophila Mulsant et Rey 1866 (Genus Rey 1866 29.Codophila varia (Fabricius 1787)Aetoloakarnania , Arkadia , Attiki , Avlis [Vahty (16.VI.1979)], Doris [Skaloula ], Evia [Agios Georgios (21.VI.1980)], Fokis [Eleon (17.VIII.1979)], Ikaria [Gyaliskari (15.VII.1981)], Ilia [Krestena (16.V.1985)], Kephallinia [Aenos Mt. (23.VI.1988)], Kynouria [Astros (5.VII.1984)], Messinia [Kazarma (4.VII.1984)], Parnassos Mt. (13.VII.1985), Rhodos , Samos , Aoos Riv. (30.V.1981), Giona (23.VII.1982), Oiti Mt. (30.IX.1984), Parnon Mt. (18.VI.1986), Vegoritis Lake (14.VII.1979). Total: 43 specimens.Dolycoris Mulsant et Rey 1866 (Genus Rey 1866 30.Dolycoris baccarum (Linnaeus 1758)Achaia [Rion (9.VI.1980)], Aetoloakarnania [Babini 23.VI.1986)], Attiki , Doris [Skaloula ], Epirus [Vryssochori (26.V.1981)], Evia [Edipsos (23.VI.1980)], Evros [Metaxades (2.VI.1982)], Florina (15.VIII.1979)], Fokis , Grevena [Anoixis (31.VII.1984)], Ilia [Kyllini (21.VII.1982)], Kephallinia [Aenos Mt. (23.VI.1988)], Konitsa [Aoos Riv. (29.V.1981)], Lakonia [Taygetos Mt. (30.IV.1985)], Messinia [Kazarma (4.VII.1984)], Kozani [Vourinos Mt. (28.V.1982)], Naxos [Moutsouna (16.VI.1981)], Oiti Mt. (30.IX.1984), Olympos Mt. , Paros [Monastirion (17.VI.1981)], Pindos , Rhodos , Rodopi Mt. , Samos [Pyrgos (14.VI.1987)], Parnon Mt. (18.VI.1986), Vourinos (28.V.1982)]. Total: 57 specimens.Dyroderes Spinola 1837 (Genus ola 1837 31.Dyroderes umbraculatus (Fabricius 1775)Doris [Skaloula (28.IV.1981)], Epirus [Vryssochori (26.V.1981)], Messinia [Dorion (1.V.1985)], Pindos [Greveniti (6.IX.1980)]. Total: 9 specimens.Eurydema Laporte 1833 (Genus rte 1833 32.Eurydema (Eurydema) eckerleini Syros [near Poseidonia (10.VI.2005)], Attiki , Tinos [near Triantaros (29.VII.2016)]. Total: 100 specimens.33.Eurydema (Horvatheurydema) fieberi, (Schummel 1837)Attiki [Kifissia (15.IV.1974)], Doris , Parnon Mt. (30.VI.1989), Kozani [Vourinos Mt. (22.VII.1989)]. Total: 5 specimens.34.Eurydema (Eurydema) oleracea (Linnaeus 1758)Creta [Rethymnon-Myloi (8.VIII.1985)], Epirus [Vryssohori (26.V.1981)], Florina , Kozani [Vourinos Mt. ], Messinia [Artemissia (29.IV.1985)], Olympos Mt. , Pindos , Rodopi Mt. , Oiti Mt. (30.IX.1984). Total: 40 specimens.35.Eurydema (Eurydema) ornata (Linnaeus 1758)Achaia [Bouboukas (21.VI.1986)], Aetoloakarnania [Mytikas-Astakos (23.VI.1986)], Arkadia [Tripolis (19.VI.1986)], Attiki , Avlis [Vathy (3.VII.1978)], Doris , Epirus [Vryssochori (27.V.1981)], Evia [Agios Georgios (21.VI.1980)], Florina [Kotas (15.VIII.1979)], Fthiotis [Malessina (21.III.1979)], Grevena [Anoixis (18.VIII.1983)], Kozani [Vourinos Mt. ], Lesvos [Sykaminea (16.VI.1987)], Parnassos Mt. [National Park (13.VII.1985)], Pieria , Rhodos , Samos [Agios Konstantinos (30.VI.1987)], Trikala [Mourgani (22.VII.1989)], Voeotia [Agia Paraskevi (8.VI.1980)], Ioannina (29.V.1981), Oiti Mt. , Parnon Mt. . Total: 75 specimens.36.Eurydema (Horvatheurydema) rugulosa (Dohrn 1861)Lesvos [Sykaminea (16.VI.1987)], Samos . Total: 6 specimens.37.Eurydema spectabilis (Stichel 1960)Crete [Agios Vassilios-Heracleon (10.VII.1985)]. Total: 12 specimens.38.Eurydema ventralis (Kolenati 1846)Doris [Marathias 1.V.1979)]. Total: 24 specimens.Eysarcoris Hahn 1834 Genus 39.Eysarcoris aeneus* (Scopoli 1763) (li 1763) Rodopi Mt. [Sidironero (23.V.1983)]. Total: 1 specimen.40.Eysarcoris ventralis Aetoloakarnania [Loutrakion-Vonitsa (22.VIII.1985)], Attiki , Crete [Rethymnon-Myloi (8.VII.1985)], Delta Acheloou (21.VII.1980), Delta Aliakmonos (19.VIII.1983), Doris , Fokis [Amfissa (19.IX.1978)], Fthiotis , Ikaria [Gyaliskari (15.VII.1981)], Ioannina , Karpathos [Arkassa (3.VI.1990)], Korinthia [Kokoni (3.VIII.1977)], Messinia [Artemissia (11.VI.1985)], Olympos Mt. [Stavros (20.V.1981)], Paros [Paroikia (18.VI.1981)], Pindos , Rhodos [Salakos (30.V.1990)], Samos , Xanthi [Porto Lagos (9.VI.1982)], Loutra Kaiafa (3.VII.1984). Total: 49 specimens.Halyomorpha Mayr 1864 Attiki , Crete [Chania (18.X.2015)], Kastellorizo [around village (15.VIII.2015)]. Total: 25 specimens.Holcogaster Fieber 1860 (Genus ber 1860 42.Holcogaster fibulata (Germar 1831)Attiki [Marathon (11.VII.1984)], Doris [Agios Nikolaos (20.VII.1986)]. Total: 2 specimens.Holcostethus Fieber 1860 Kynouria [Astros (31.VII.1984)]. Total: 1 specimen.44.Holcostethus sphacelatus (Fabricius 1794)Epirus [Vryssochori (26.V.1981)], Konitsa [Aoos Riv. 29.V.1981)], Olympos Mt. [Prionia (21.V.1981)], Rodopi Mt. [Virgin Wood 23.V.1983)]. Total: 7 specimens.Mustha Amyot et Serville 1843 (Genus lle 1843 45.Mustha spinulosa (Lefebvre 1831)Attiki , Doris [Skaloula (18.V.1989)], Trikala [Mourgani (19.VI.1985)]. Total: 6 specimens.Neostrachia Dallas 1851 Genus 46.Neostrachia bisignata Doris [Monastirakion (29.VII.1982)], Kynouria [Astros (24.VIII.1988)], Volos [Amaliapolis (1.IX.1990)]. Total: 3 specimens.Neottiglossa Kirby 1837 Genus 47.Neottiglossa bifida (Costa 1847)Achaia [Bouboukas (21.VI.1986)], Aetoloakarnania , Doris , Messinia , Konitsa [Aoos-Monastiri (30.V.1981)], Lakonia [Mystras (30.IV.1985)], Naxos [Apeiranthos (16.VI.1981)], Pieria [Poroi (14.VIII.1980)], Thessaloniki [Plagiarion (23.V.1981)]. Total: 22 specimens.48.Neottiglossa flavomarginata (Lucas 1849)Kozani [Vourinos Mt. (29.VI.1984)]. Total: 6 specimens.49.Neottiglossa leporina (Herrich-Schaeffer 1830)Doris , Drama [Potami (12.VI.1982)], Epirus [Vryssochori (26.V.1981)], Florina , Konitsa [Aoos-Monastiri (10.V.1981)], Kozani [Vourinos Mt. (8.VI.1984)], Pieria [Poroi (14.VIII.1980)], Rodopi Mt. , Oiti Mt. ]. Total: 22 specimens.50.Neottiglossa lineolata* (Mulsant et Rey 1852) (ey 1852) Doris [Skaloula (18.VII.1978)], Evia , Konitsa [Aoos-Monastiri (30.V.1981)], Lesvos [Antissa (17.VI.1987)], Xanthi [Porto Lagos (9.VI.1982)]. Total: 8 specimens.51.Neottiglossa pusilla* (en 1789) Rodopi Mt. [Vathyrhemma (26.VII.1982)]. Total: 2 specimens.Nezara Amyot et Serville 1843 (Genus lle 1843 52.Nezara viridula (Linnaeus 1758)Attiki . Crete [Heracleon-Agios Vassilios (10.VII.1985)], Paros [Paroikia (18.IX.1981)]. Total: 13 specimens.Palomena Mulsant et Rey 1866 Kozani [Vourinos Mt. (28.V.1982)], Oiti Mt. (30.IX.1984), Parnon Mt. [Kastanitsa ], Pindos [Miliotades (29.IX.1980)], Rodopi Mt. [Betula Forest (22.V.1983)], Tripolis [Tripolis (19.VI.1986)]. Total: 10 specimens.Pentatoma Olivier 1789 (Genus ier 1789 54.Pentatoma (Pentatoma) rufipes (Linnaeus 1758)Florina , Rodopi Mt. . Total: 10 specimens.Peribalus Mulsant et Rey, 1866 Attiki , Avlis [Vathy (3.VII.1978)], Crete [Agios Vassilios-Heracleon (25.IV.1975)], Doris [Skaloula ], Florina [Kotas (15.VIII.1979)], Fthiotis [Malessina (21.III.1979)], Parnassos Mt. [National Park (13.VII.1985)], Rhodos [Salakos (30.V.1990)], Samos [Agios Konstantinos (27.VI.1987)]. Total: 16 specimens.Piezodorus Fieber 1861 (Genus ber 1861 56.Piezodorus lituratus (Fabricius 1794)Attiki [Parnis-Mola (12.VII.1985)], Schinias [(16.V.1984)], Avlis [Vathy 13.IV.1978)], Doris , Epirus [Vryssochori (26.V.1981)], Evia [Agios Georgios (21.VI.1980)], Evrytania [Megalo Chorio (10.VIII.1986)], Florina [Vernon Mt.-Kalo Nero (30.VII.1982)], Messinia [Menina (30.IV.1985)], Ilia [Killini Mt. (21.VII.1982)], Kerkyra [Tritsi (3.IV.1985)], Oaks [Rodopi (12.VIII.1985)], Parnon Mt., [Kastanitsa ], Ikaria [Raches (14.VII.1981)], Rhodos . Total: 31 specimens.Rhaphigaster Laporte 1833 (Genus rte 1833 57.Rhaphigaster nebulosa (Poda 1761)Acheloos Riv. [Delta Acheloou (21.VII.1990)], Attiki , Avlis [Vathy (5.V.1981)], Crete [Heracleon-Agios Vassilios (10.VII.1985)], Doris , Evros Riv. [Delta Evrou (7.VI.1982)], Aetoloakarnania [Nafpaktos (4.IX.1979)], Rhodos [Petaloudes (29.V.1990)]. Total: 17 specimens.Sciocoris Fallen 1829 (Genus len 1829 58.Sciocoris (Sciocoris) cursitans cursitans (Fabricius 1794)Doris [Skaloula (12.IV.1985)], Lesvos [Andissa (17.VI.1987)]. Total: 2 specimens.59.Sciocoris (Sciocoris) deltocephalus (Fieber 1861)Kephallinia [Aenos Mt. (23.VI.1988)], Paros [Paroikia (18.VI.1981)]. Total: 2 specimens.60.Sciocoris (Sciocoris) helferii (Fieber 1851)Aetoloakarnania [Mytikas-Astakos (23.VI.1986)], Arkadia [Xeropigado (9.V.1985)], Attiki [Parnis Mt.-Mola (12.VII.1985)], Ikaria [Gialiskari (15.VII.1981)], Kephallinia [Aenos Mt. (23.VI.1988)], Konitsa [Aoos-Monastiri (30.V.1981)], Kozani [Vourinos Mt. (29.VI.1984)], Aoos Riv. (14.VIII.1986)], Lesvos [Kalloni (18.VI.1987)], Pieria [Varikon (23.VII.1982)]. Total: 11 specimens.61.Sciocoris (Aposciocoris) macrocephalus (Fieber 1851)Arkadia [Xeropigado (9.V.1985)], Attiki [Avlon Marathon ], Doris [Skaloula (1.VI.1985)], Fokis [Amfissa (2.V.1979)], Kephallinia [K. Katelios (22.VI.1988)], Kerkyra [Liapades (2.IV.1985)]. Total: 16 specimens.62.Sciocoris (Neosciocoris) maculatus (Fieber 1851)Kephallinia [Aenos Mt. (23.VI.1988)], Konitsa , Olympos Mt. [Kryovryssi (30.V.1982)], Samothraki [Loutra (4.VI.1982)], Oiti Mt. (9.VIII.1986). Total: 8 specimens.63.Sciocoris (Sciocoris) sulcatus (Fieber 1851)Aetoloakarnania [Messolongion-Tourlida (20.VII.1990)], Attiki , Kephallinia [Aenos Mt. (23.VI.1986)], Korinthia [Derveni (15.V.1985)], Kozani [Vourinos Mt. (29.VI.1984)], Kynouria [Kastanitsa (20.V.1982)], Messinia [Dorion (1.V.1985)], Olympos Mt. [Prionia (21.V.1981)], Pindos [Milies (20.VIII.1985)], Rhodos , Rodopi Mt. [Elatia (25.VII.1982)], Voeotia [Tsoukalades (12.V.1988)]. Total: 19 specimens.Stagonomus Gorski 1852 (Genus ski 1852 64.Stagonomus (Stagonomus) amoenus (Brulle 1832)Crete [Heracleon-Agios Vassilios (25.IV.1985)], Konitsa , Paros [Monastirion (17.VI.1981)], Rhodos [Petaloudes (29.V.1990)]. Total: 15 specimens.65.Stagonomus bipunctatus (Linnaeus 1758)Evia [Agios Georgios (21.VI.1980)], Florina [Vermon Mt.-Kalo Nero (30.VII.1982)], Kozani [Vourinos Mt. (18.VIII.1985)]. Total: 7 specimens.Staria Dohrn 1860 (Genus hrn 1860 66.Staria lunata (Hahn 1835)Arkadia [Xeropigado (9.V.1985)], Attiki [Marathon (11.VII.1984)], Doris [Skaloula ], Evros [Metaxades (2.VI.1982)], Fokida [Agia Euthymia (12.V.1988)], Konitsa [Aoos (27.V.1981)], Korinthia [Akrokorinthos (31.VII.1979)], Kozani [Vourinos Mt. (28.VI.1984)] Menalon Mt. (20.VI.1986), Parnassos Mt. , Rodopi Mt. [Vathyrhemma (26.VII.1982)]. Total: 36 specimens.Subfamily PodopinaeAncyrosoma Amyot et Serville 1843 (Genus lle 1843 67.Ancyrosoma leucogrammes (Gmelin 1790)Aetoloakarnania , Arkadia [Xeropigado (9.V.1985)], Arta [Louros Riv. (28.IX.1981)], Attiki , Avlis [Vathy (16.VI.1979)], Doris , Evia [Agios Georgios (21.VI.1980)], Fokis (Agia Euthymia (12.V.1988)], Ioannina (29.V.1981), Karditsa [Artessiano (1.X.1981)], Konitsa [Aoos Riv. ], Korinthia , Kynouria [Astros ], Messinia [Kazarma (4.VII.1984)], Olympos Mt. [Prionia (13.VIII.1979)], Pieria [Poroi (14.VIII.1980)], Platanos [Parnon Mt. (1.VII.1985)], Thessaloniki [Cedron Hills (23.V.1981)]. Total: 57 specimens.Derula Mulsant et Rey 1856 (Genus Rey 1856 68.Derula flavoguttata (Mulsant et Rey 1856)Attiki [Schinias (23.VI.1986)], Doris , Epirus [Vryssochori (26.V.1981)], Evia , Florina , Fokis [Agia Euthymia (12.V.1988)], Korinthia [Derveni (15.V.1985)], Pindos [Vouchorina (13.V.1983)], Rhodos [Salakos (30.V.1990)], Samos [Pythagorion (23.VI.1987)], Trikala [Mourgani (18.VI.1985)]. Total: 23 specimens.Graphosoma Laporte 1833 Aetoloakarnania [Aetolikon (20.VII.1990)], Attiki , Crete [Rethymnon-Myloi (8.VII.1985)], Doris [Skaloula ], Evrytania [Megalo Chorio (10.VIII.1986)], Fokis [Amfissa (8.VI.1980)] Ioannina [Vikos Gorge (13.VIII.1986)], Konitsa [Aoos-Monastiri (3.V.1981)], Korinthia [Derveni (15.V.1985)], Kozani , Lakonia [Mystras (30.IV.1986)], Lesvos [Sykaminea (16.VI.1987)], Olympos Mt. [Stavros (13.VIII.1980)], Pieria [Litochoron (13.VIII.1979)], Rhodos [Apolakkia (30.VI.1987)], Voeotia [Aliartos (12.V.1988)], Oiti Mt. . Total: 28 specimens.70.Graphosoma semipunctatum (Fabricius 1775)Attiki , Crete [Rethymnon-Agioi Apostoloi (30.VI.1993)], Doris [Skaloula (18.VI.1978)], Kephallinia [Myrtos (25.VI.1988)], Lakonia [Mystras (11.VI.1985)], Oidi Mt. (27.VI.1993), Samos [Neochorion (14.VI.1987)]. Total: 23 specimens.Leprosoma Baerensprung 1859 (Genus ung 1859 71.Leprosoma inconspicuum (Baerensprung 1859)Doris [Kalion (5.V.1975)]. Total: 1 specimen.Podops Laporte 1833 Genus 72.Podops (Opocrates) curvidens (Costa 1843)Aetoloakarnania , Doris , Pieria [Varikon (23.VII.1982)], Pindos . Total: 14 specimens.73.Podops (Podops) inunctus* (Fabricius 1775) (us 1775) Doris [Kalion (15.VII.1978)], Samothraki [Kamariotissa (5.VI.1982)]. Total: 4 specimens.74.Podops (Opocrates) rectidens Horvath 1883Achaia [Bouboukas (9.VI.1980)], Aetoloakarnania [Mytikas-Astakos (23.VI.1986)], Arkadia [Kynouria-Astros ], Attiki [Schinias (1.VII.1982)], Doris , Ikaria [Gialiskari (15.VII.1981)], Ilia [Pyrgos-Zacharo (3.VII.1984)], Samothraki [Kamariotissa (5.VI.1982)], Zakynthos [L. Keri (29.VI.1988)]. Total: 23 specimens.Tarisa Amyot et Serville 1843 Genus 75.Tarisa pallescens* (Jakovlev 1871) (ev 1871) Olympos Mt. [Kryovryssi (30.V.1982)]. Total: 1 specimen.Tholagmus St\u00e5l 1860 (Genus t\u00e5l 1860 76.Tholagmus flavolineatus (Fabricius 1798)Arkadia [Charadros (17.VI.1986)], Attiki [Avlon ], Avlis [Vathy (16.VI.1979)], Evia [Agios Georgios (21.VI.1980)], Fthiotis [Kalamakion (18.VIII.1979)], Kephallinia , Aetoloakarnania [Kapsorachi-Trichonis (22.VI.1986)], Menalon Mt. (20.VI.1986), Parnassos Mt. [National Park (13.VII.1985)]. Total: 18 specimens.Ventocoris Hahn 1843 (Genus ahn 1843 77.Ventocoris (Selenodera) achivus (Horvath 1889)Avlis [Vathy (16.VI.1979)]. Total: 2 specimens.78.Ventocoris (Ventocoris) rusticus (Fabricius 1781)Achaia [Kato Klitoria (20.VI.1986)], Attiki [Marathon (14.V.1985)], Avlis [Vathy ], Evia [Agios Georgios (21.VI.1980)], Fokis [Amphissa (8.VI.1980)], Kephallinia [Assos (25.VI.1988)], Messinia [Kazarma (4.VII.1984)], Santorini Island [Kamarion (5.V.1982)], Voeotia [Agia Paraskevi (23.VII.1980)]. Total: 23 specimens.Family PLATASPIDAESubfamily PlataspinaeCoptosoma Laporte 1833 (Genus rte 1833 79.Coptosoma scutellatum (Geoffroy 1785)Achaia [Bouboukas (21.VI.1986)], Doris , Florina [Vernon Mt.-Kalo Nero (21.VII.1983)], Ioannina [Voutsaras (24.VI.1984)], Kozani [Vourinos Mt. (18.VIII.1985)], Olympos Mt. [Prionia (13.VIII.1980)], Parnassos Mt. [National Park (13.VII.1985)], Pindos [Korydallos (25.V.1981)], Prespa [Bela Voda (13.X.1986)], Rodopi [Vathyrhemma (26.VII.1982)], Aoos Riv. (27.V.1981). Total: 32 specimens.Family SCUTELLERIDAESubfamily EurygastrinaeEurygaster Laporte 1833 Genus 80.Eurygaster austriaca (Schrank 1778)Avlis [Vathy ], Fokis [Galaxidi (15.IV.1980)], Oiti Mt. (1.VII.1984). Total: 4 specimens.81.Eurygaster dilaticollis (Dohrn 1860)Rodopi Mt. (19.VII.1983), Oiti Mt. , Vernon Mt. [Kalo Nero (30.VI.1982)]. Total: 6 specimens.82.Eurygaster hottentotta* (Fabricius 1775) (us 1775) Kynouria [Kastanitsa (20.VII.1982)]. Total: 1 specimen.83.Eurygaster integriceps (Puton 1881)Arkadia [Xeropigado (9.V.1985)], Attiki [Marathon (14.V.1978)], Avlis [Vathy (13.IV.1978)], Chios , Doris [Skaloula ], Epirus [Vryssochori (26.V.1981)], Grevena , Ikaria [Raches (14.VII.1981)], Ioannina (29.V.1981), Kozani [Vourinos Mt. (29.VI.1984)], Kynouria [Astros (20.V.1982)], Naxos [Apeiranthos (16.VI.1981)], Paros [Paroikia (18.VI.1981)], Samos . Total: 31 specimens.84.Eurygaster maura (Linnaeus 1758)Doris , Epirus [Vryssochori (27.V.1981)], Evia [Agios Georgios (21.VI.1980)], Florina , Kastoria (13.V.1983), Kynouria [Astros ], Messinia [Kazarma (4.VII.1984)], Pindos [Korydallos (25.V.1981)], Rhodos , Rodopi Mt. , Xanthi [Porto Lagos (8.V.1982)], Zakynthos [Keri (29.VI.1988)]. Total: 21 specimens.85.Eurygaster testudinaria testudinaria (Geoffroy 1785)Kastoria (13.V.1983), Rodopi Mt. [Silli (23.VIII.1983)]. Total: 3 specimens.Psacasta Germar 1839 (Genus mar 1839 86.Psacasta (Psacasta) exanthematica exanthematica (Scopoli 1763)Aetoloakarnania [Nafpaktos (22.VII.1978)]. Total: 1 specimen.Subfamily OdontoscelinaeIrochrotus Amyot et Serville 1843 (Genus lle 1843 87.Irochrotus maculiventris (Germar 1839)Achaia [Bouboukas (21.VI.1986)], Arkadia [Tripolis (19.VI.1986)], Attiki [Avlon (26.V.1978)], Doris [Skaloula (1.VI.1985)], Evia , Ioannina [Voutsaras (24.VI.1984)]. Total: 8 specimens.Odontoscelis Laporte 1833 (Genus rte 1833 88.Odontoscelis (Odontoscelis) fuliginosa (Linnaeus 1761)Attiki , Karpathos [Messochori (4.VI.1990)], Kephallinia [Aenos Mt. (23.VI.1988)]. Total: 7 specimens.89.Odontoscelis (Odontoscelis) lineola (Rambur 1839)Doris [Skaloula (17.IX.1978)] Arkadia , Vegoritis Lake (14.VIII.1979). Total: 4 specimens.Subfamily OdontotarsinaeOdontotarsus Laporte 1833 (Genus rte 1833 90.Odontotarsus purpureolineatus (Rossi 1790)Achaia [Bouboukas (21.VI.1986)], Aetoloakarnania , Arkadia [Charadros (17.VI.1986)], Attiki , Chios [Armolia (20.VI.1987)], Doris [Skaloula (20.VII.1986)], Epirus [Vryssochori (26.V.1981)], Evros [Metaxades (2.VI.1982)], Grevena [Anoixis (31.VII.1984)], Ilia [Kyllini Mt. (21.VII.1982)], Ioannina [Voutsaras (24.VI.1984)], Konitsa [Aoos Riv.-Monastiri (29.V.1981)], Kozani , Lesvos [Petra (17.VI.1987)], Messinia , Olympos Mt. [Prionia (21.V.1981)], Pilion Mt. [Chania 16.VIII.1980)], Rhodos , Samos , Trikala [Mourgani (19.VI.1985)], Voeotia [Tsoukalades (12.V.1988)], Parnon Mt. (11.VI.1985). Total: 36 specimens.91.Odontotarsus robustus (Jakovlev 1883)Attiki , Doris [Skaloula ], Pindos . Total: 16 specimens.92.Odontotarsus rufescens (Fieber 1861)Amphilochia [Anoixiatiko (20.VIII.1985)], Attiki [Avlon (20.V.1987)], Evia [Agios Georgios (21.VI.1980)], Ilia [Krestena (16.V.1985)], Kynouria [Astros (13.VII.1982)], Messinia [Kazarma (4.VII.1984)]. Total: 7 specimens.In Greece, until the present work, very few studies have been conducted on the Pentatomoidea fauna, in which 150 Pentatomoidea species are referred. Except two overall studies performed by Drosopoulos and Rams"} +{"text": "J Clin Invest. 2022;132(8):e149160. https://doi.org/10.1172/JCI149160Original citation: J Clin Invest. 2023;133(3):e168441. https://doi.org/10.1172/JCI168441Citation for this corrigendum: The left panel of The authors regret the error."} +{"text": "Phil. Trans. R. Soc. B377, 20210159. (Published online 2 May 2022). (https://doi.org/10.1098/rstb.2021.0159)In the original version of this article, author Pengjuan Zu's name was spelled incorrectly.This has now been corrected on the publisher's website."} +{"text": "Consent was added to this article: We have informed consent statements from all patients.125-130. https://doi.org/10.18632/oncotarget.27854Original article: Oncotarget. 2021; 12:125\u2013130."} +{"text": "Investig. Ophthalmol. Vis. Sci.25, 463 (1984).Screening for color blindness using optokinetic nystagmusThe correct reference is listed below:Investigative ophthalmology & visual science\u00a025, 463\u2013466 (1984).Cavanagh, P., Anstis, S. & Mather, G. Screening for color blindness using optokinetic nystagmus. The original Article has been corrected."} +{"text": "This article has been corrected: In 61703-61715. https://doi.org/10.18632/oncotarget.11225Original article: Oncotarget. 2016; 7:61703\u201361715."} +{"text": "The erratum corrects an error in Fig. 4 of the published article. J. Biomed. Opt.27(7), 074705 (2022) doi: https://doi.org/10.1117/1.JBO.27.7.074705 was originally published on 28 October 2022 with an error in Fig. 4. The axes were reversed in the original version:This article [Corrected version:The error was corrected on 9 July 2022."} +{"text": "JCI Insight. 2020;5(19):e139932. https://doi.org/10.1172/jci.insight.139932Original citation: JCI Insight. 2022;7(20):e165502. https://doi.org/10.1172/jci.insight.165502Citation for this corrigendum: The authors recently became award of inconsistencies in The authors regret the errors."} +{"text": "First published: 03 November 2022https://doi.org/10.1111/jvim.16554Volume 36, Issue 6Correction to reference number 13 as follows. The last name of Jelinicic does not use diacritics.Inst Safe Medicat Pract. 2004; 15(1): 10\u201011.13. Koczmara C, Hyland S, Jelincic V. High alert: preventing insulin errors."} +{"text": "JCI Insight. 2021;6(4):e140180. https://doi.org/10.1172/jci.insight.140180Original citation: JCI Insight. 2022;7(23):e167011. https://doi.org/10.1172/jci.insight.167011Citation for this corrigendum: Following the publication of this article, the authors became aware of similarities between the CYP2E1 blot in The authors regret the error."} +{"text": "Consent was added to this article: The patient mentioned in the paper signed the informed consent statement on 13/10/2020.1116-1121. https://doi.org/10.18632/oncotarget.27966Original article: Oncotarget. 2021; 12:1116\u20131121."} +{"text": "This article has been corrected: In 2543-2559. https://doi.org/10.18632/oncotarget.27650Original article: Oncotarget. 2020; 11:2543\u20132559."} +{"text": "J Clin Invest. 2020;130(7):3684\u20133698. https://doi.org/10.1172/JCI136908Original citation: J Clin Invest. 2023;133(5):e169500. https://doi.org/10.1172/JCI169500Citation for this corrigendum: In the original version of The authors regret the error."} +{"text": "Computational simulation of flow-induced arterial remodeling of the pancreaticoduodenal arcade associated with celiac artery stenosis. Journal of Biomechanics. 2019;92:146-154.https://doi.org/10.1016/j.jbiomech.2019.05.043"} +{"text": "JCI Insight. 2020;5(17):e133225. https://doi.org/10.1172/jci.insight.133225Original citation: JCI Insight. 2022;7(20):e165688. https://doi.org/10.1172/jci.insight.165688Citation for this corrigendum: The authors recently became aware that one of the p-TBK1 blot images presented in The authors regret the error."} +{"text": "In the originally published version of the below-listed manuscripts, the html version incorrectly lists the article editor as an author.These errors have been corrected. The publisher apologizes for the errors.https://doi.org/10.1093/jmcb/mjab076https://doi.org/10.1093/jmcb/mjab078https://doi.org/10.1093/jmcb/mjab079https://doi.org/10.1093/jmcb/mjab081https://doi.org/10.1093/jmcb/mjab082https://doi.org/10.1093/jmcb/mjac002"} +{"text": "J. R. Soc. Interface14, 20170247. (Published online 31 May 2022) (https://doi.org/10.1098/rsif.2017.0247)This corrigendum corrects the record and includes Mahesh M. Bandi and Madhusudhan Venkadesan as corresponding authors for the article.bandi@oist.jpm.venkadesan@yale.edushreyas.mandre@warwick.ac.uk"} +{"text": "Correction:\u00a0International Journal of Implant Dentistry\u00a0(2022) 8:43https://doi.org/10.1186/s40729-022-00445-zIncorrect: C. B. SpiesCorrect: B. C. SpiesThe original publication of this article containe"} +{"text": "This article has an addendum: Patient consent was obtained from the patient.81969-81971. https://doi.org/10.18632/oncotarget.13046Original article: Oncotarget. 2016; 7:81969\u201381971."} +{"text": "Following the publication of the original article , we wereOriginally published name: Raghda R. S. RoshdyCorrect name: Raghda R. S. HusseinThe original article has been corrected."} +{"text": "JCI Insight. 2022;7(15):e160989. https://doi.org/10.1172/jci.insight.160989Original citation: JCI Insight. 2022;7(18):e164813. https://doi.org/10.1172/jci.insight.164813Citation for this corrigendum: For clarity, the authors have updated The authors regret the errors."} +{"text": "Correction: BMC Psychiatry. 20, 229 (2020)https://doi.org/10.1186/s12888-020-02596-yFollowing the publication of the original article , the autThe incorrect code of the study is: IR.KUMS.REC.1397.187The correct code of the study is: IR.KUMS.REC.1397.866The original article has been"} +{"text": "In the originally published version of the below-listed manuscripts the data availability statement was accidentally omitted. Data availability statements have now been added to the following papers.https://doi.org/10.1093/ehjdh/ztaa012https://doi.org/10.1093/ehjdh/ztaa018https://doi.org/10.1093/ehjdh/ztab009https://doi.org/10.1093/ehjdh/ztab011https://doi.org/10.1093/ehjdh/ztab032https://doi.org/10.1093/ehjdh/ztab034https://doi.org/10.1093/ehjdh/ztab038https://doi.org/10.1093/ehjdh/ztab051https://doi.org/10.1093/ehjdh/ztab082https://doi.org/10.1093/ehjdh/ztab108"} +{"text": "J Clin Invest. 2016;126(5):1783\u20131800. https://doi.org/10.1172/JCI83669Original citation: J Clin Invest. 2023;133(4):e169304. https://doi.org/10.1172/JCI169304Citation for this corrigendum: 2O and WT/DSS pictures were inadvertently switched. In \u2013/\u2013Ptpn22/DSS sample. The correct figure is shown below.The authors recently became aware of errors in The authors regret the errors."} +{"text": "This article has an addendum: The patient has previously consented to publication of de-identified findings.4195-4200. https://doi.org/10.18632/oncotarget.27793Original article: Oncotarget. 2020; 11:4195\u20134200."} +{"text": "Oncogene 10.1038/onc.2013.594, published online 27 January 2014Correction to: Supplementary figure The CDC42 blot in Suppl. fig The corrected figure is given below.Supplementary Figure 1"} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-022-24228-z, published online 16 November 2022Correction to: The original version of this Article contained an error in Reference 10, which was incorrectly given as:635, 413833. (2022).Das, S. Particle scattering by harmonically trapped quantum gases in an artificial magnetic field. Phys. B Condens. Matter The correct reference is listed below:Phys. B Condens. Matter635, 413833. https://doi.org/10.1016/j.physb.2022.413833 (2022).Das, S. Particle scattering by harmonically trapped quantum gases in an artificial magnetic field. The original Article has been corrected."} +{"text": "PNAS Nexus was opened, it was incorrectly labeled March 2023. The month has been corrected, and citation details of the following list of articles were affected by this error:When Volume 2, Issue 1, January 2023 of https://doi.org/10.1093/pnasnexus/pgac306Cystic fibrosis rabbits develop spontaneous hepatobiliary lesions and CF-associated liver disease (CFLD)-like phenotypes. https://doi.org/10.1093/pnasnexus/pgac298High-resolution cryo-EM structure of the junction region of the native cardiac thin filament in relaxed state. https://doi.org/10.1093/pnasnexus/pgac297Topical SCD-153, a 4-methyl itaconate prodrug, for the treatment of alopecia areata. https://doi.org/10.1093/pnasnexus/pgac300Development of an in-vitro high-throughput screening system to identify modulators of genitalia development. https://doi.org/10.1093/pnasnexus/pgac2902,3,7,8-Tetrachlorodibenzo-p-dioxin induces multigenerational alterations in the expression of microRNA in the thymus through epigenetic modifications. Wolbachia. https://doi.org/10.1093/pnasnexus/pgac293Cell-based analysis reveals that sex-determining gene signals in Ostrinia are pivotally changed by male-killing https://doi.org/10.1093/pnasnexus/pgac286Information maximization explains state-dependent synaptic plasticity and memory reorganization during non-rapid eye movement sleep. https://doi.org/10.1093/pnasnexus/pgac288Pharmacological potentiators of the calcium signaling cascade identified by high-throughput screening. https://doi.org/10.1093/pnasnexus/pgac285Leaded aviation gasoline exposure risk and child blood lead levels. https://doi.org/10.1093/pnasnexus/pgac283On the interplay of hierarchies, conflicts, and cooperation: An experimental approach. https://doi.org/10.1093/pnasnexus/pgac270Senescent stroma induces nuclear deformations in cancer cells via the inhibition of RhoA/ROCK/myosin II-based cytoskeletal tension. https://doi.org/10.1093/pnasnexus/pgac292On shape forming by contractile filaments in the surface of growing tissues. https://doi.org/10.1093/pnasnexus/pgac291Customizable, reconfigurable, and anatomically coordinated large-area, high-density electromyography from drawn-on-skin electrode arrays. https://doi.org/10.1093/pnasnexus/pgac295Watershed memory amplified the Oroville rain-on-snow flood of February 2017. https://doi.org/10.1093/pnasnexus/pgac275A deep-learning model of prescient ideas demonstrates that they emerge from the periphery. https://doi.org/10.1093/pnasnexus/pgac289Risky cascading transitions in international relationships. https://doi.org/10.1093/pnasnexus/pgac281Moralized language predicts hate speech on social media. The citation details for each of the above articles have been updated to reflect the correct month of this issue. The Publisher regrets this error."} +{"text": "Correction to: bord personal disord emot dysregul 8, 28 (2021).https://doi.org/10.1186/s40479-021-00169-3Following publication of this article , it is rMaria Lidia Gerra.Martina Ardizzi.Silvia Martorana.Veronica Leoni.Paolo Riva.Emanuele Preti.Barbara Francesca Marta Marino.Paolo Ossola.Carlo Marchesi.Vittorio Gallese.Chiara De Panfilis.The original article has been updated."} +{"text": "This article has been corrected: Due to errors during figure assembly, the GAPDH blots in 8822-8838. https://doi.org/10.18632/oncotarget.3558Original article: Oncotarget. 2015; 6:8822\u20138838."} +{"text": "This article has been corrected: Due to errors during the assembly of 23401-23413. https://doi.org/10.18632/oncotarget.15581Original article: Oncotarget. 2017; 8:23401\u201323413."} +{"text": "There is an error in reference 24. The correct reference is: Diamond Project Group. Incidence and trends of childhood Type 1 diabetes worldwide 1990\u20131999. Diabet Med. 2006;23(8):857\u201366. Epub 2006/08/17. pmid:16911623."} +{"text": "Retraction Note to: Gene Therapy10.1038/gt.2011.82.The editor has retracted this article followin"} +{"text": "JCI Insight. 2017;2(16):e93076. https://doi.org/10.1172/jci.insight.93076Original citation: JCI Insight. 2021;6(7):e149895. https://doi.org/10.1172/jci.insight.149895Citation for this corrigendum: NHBA Control). An institutional review committee concluded that the error in The Editors previously posted an Expression of Concern for this article regarding images in The authors regret the error."} +{"text": "This article has been corrected: In 62425-62438. https://doi.org/10.18632/oncotarget.11515Original article: Oncotarget. 2016; 7:62425\u201362438."} +{"text": "Correction to:Mucosal Immunology (2017); advance online publication, 23 August 2017; doi:10.1038/mi.2017.68Panel f of The publisher regrets the error."} +{"text": "This article has been corrected: The bioinformatics analysis of TCGA data in 31785-31801. https://doi.org/10.18632/oncotarget.15991Original article: Oncotarget. 2017; 8:31785\u201331801."} +{"text": "JCI Insight. 2021;6(6):e135753. https://doi.org/10.1172/jci.insight.135753Original citation: JCI Insight. 2021;6(8):e150137. https://doi.org/10.1172/jci.insight.150137Citation for this corrigendum: During the preparation of this manuscript, the center panel of the bottom row of Figure 2D was inadvertently duplicated from the left panel of the bottom row of Figure 2D. The figure has been corrected.The authors regret the error."} +{"text": "This article has been corrected: During reorganization of 1606-1624. https://doi.org/10.18632/oncotarget.26600Original article: Oncotarget. 2019; 10:1606\u20131624."} +{"text": "JAC-Antimicrobial Resistance 2021; https://doi.org/10.1093/jacamr/dlab051JAC-Antimicrobial Resistance 2021; https://doi.org/10.1093/jacamr/dlab052JAC-Antimicrobial Resistance 2021; https://doi.org/10.1093/jacamr/dlab053JAC-Antimicrobial Resistance 2021; https://doi.org/10.1093/jacamr/dlab054JAC-Antimicrobial Resistance 2021; https://doi.org/10.1093/jacamr/dlab055JAC-Antimicrobial Resistance 2021; https://doi.org/10.1093/jacamr/dlab056In the original published versions of these articles, the link to the prescribing information for cefiderocol was missing. This has now been corrected in the published versions of the articles. The authors apologize for this error."} +{"text": "Correction to: Trials 18, 528 (2017)https://doi.org/10.1186/s13063-017-2270-3Originally published name: Rosanna W. L. LamCorrected name: Rosanna W. L. LauFollowing the publication of the original article , we wereThe original article has been corrected."} +{"text": "Global Medical Geneticswishes to recognize those who contributed as an expert peer-reviewer of submitted scientific papers in 2021.Global Medical Genetics.Thank you for your contributions toIwona \u017burTomer Ziv-BaranAndrea ZiniQian-Hao ZhuJiang ZhouYingmei ZhangXinjiang Zhang\u00c1ngel Zarain-HerzbergSarah E. ZandersHakimeh ZaliRaymond YungYonggui YuanJin-qing YuanHao YuChao YuPierre YouinouYeliz YolVa yoT. Ylisaukko-ojaHiroyuki YasudaBing YaoJunichi YamaguchiSuowen XuClaudia WylezichYah-Huei Wu-ChouWei-Te WuJianghong WuJiande WuKris C. WoodHye Ryun WooJongkonnee WongpiyabovornLoo Keat WeiJill WegrzynMilind WatveZehua WangXiaojuan WangWeiping WangIoannis A. VoutsadakisVandana VinayakRafael Velazquez-CruzAB VedamurthySarinnapha M. VasunilashornSirisha VammiZehra Oya UygunerAsta TvarijonaviciuteDilsad TurkdoganP\u0131nar TulayAgeliki TsagaratouAnthony TrewavasVan Khanh TranAnn-Marie TorregrossaHirofumi TomitaLucia TombolanPascale TomasiniMaria TomasPavel TomancakA. TirellaPilar S. TestillanoLeroy ten DamDuygu TekinEleni TaniYuki TamuraMurtaza M. TambuwalaGerasimos P. SykiotisZ. Renee SungLi-Peng SunAihua SunTakaaki SugikiIrna SufiawatiJose Suazowenling SUChun-Li SuMaja StojiljkovicAgnieszka StembalskaElizabeth K. SpeliotesKayal DM SmitaRobert SmigielMichael K. SkinnerSanjay M. SisodiyaDavid SimarByoung-Soo ShinJin-Yuan ShihTakahiko Shibaharapei ShiBrian M. ShewchukLisha ShenPil Joon SeoAida Semic-JusufagicBarry ScottRobert J. SchmitzMari SandellMaria, de la Paz SanchezEmine Turkmen SamdanciLynn Y. SakaiOzlem SaglamReza SafaralizadehEnrique SaezAmets SaenzAlexandre RouenVeronique RossiH. Llewelyn RoderickKate E. RileyBergmann M. RibeiroSilvia R. A. ReisTheodore P. RasmussenPragna RaoAna Lucia Carrinho Ayroza RangelBhama RamkhelawonElizabeth J. RadfordLarysa Y. PylypAlkes L. PriceShikha PrasadPranav Kumar PrabhakarN. Pop-JordanovaLydia PooleS. R. PhadkePiero PeruccaA. M. PersicoPascale PerrinIsrael Perez-TorresA. Pena-RosadoA. PellicerAles PecinkaSmaranika PattnaikAlireza PasdarAlireza PasdarSeppo ParkkilaHumberto, Jr. ParadaPier Francesco PalamaraBulent OzpolatJuan M. OrdunaM. OliveAnna B. OhlssonB. NowickaParisa NorouzitallabZahra NoormohammadiTakahiro NobuzaneMasha Y. NivDuc Hinh NguyenYun Fong NgeowPraveen Kumar NeelaHashem NayeriSatoshi NarumiMasataka Narukawajuan NanGerson NakazatoA. J. MunozMridul MukherjiSrabani MukherjeeHirohito MiuraMilica MiljkovicChiara MilaniL. MiglioreJos\u00e9 Luis MicolJames J. MezhirPeter MeyerLisa MethvenJan A. MennigenShaban MelI.M. Medina-D\u00edazKatsuhiro MasagoEfr\u00e9n Mart\u00ednez-QuintanaMarcella MartinelliLucia MargariChao MaoGaia Chiara ManninoLicinio MancoJos\u00e9 E. ManautouKristiina M\u00e4kinenAbdolkarim MahroozMagdy M. MahfouzAbbas Ali MahdiAsri MaharaniPaul S. MaddoxMike MacknessRenato Assis MachadoMalou A. LugthartVito LongoHanns LochmullerXian-zhi LiuShoufeng LIUCharlotte LingJinpiao LinAnatoly V. LichtensteinMarc LibaultWilson LiaoEvi LianidouXiaomin;Xiaowan Li;Lixiaomin;juan li;heXiajun LiAnna-Liisa LevonenTomoko LeeSeung Jin LeeSeong-Wook LeeJefferson LEEM. A. LebedevaDouglas A. LauffenburgerJunko KusumiSetor K. KunutsorAlexei P. KudinKsenia V. KrasilevaClaudia K\u00f6hlerDo-Hyung KimAndre Salim KhayatNaim A. KhanDr. Mahamad Irfanulla KhanGolsa KetabchiMohammad Amin KerachianKathleen L. KellerFuminori KawabataFuminori KawabataUshang V. KateJason KaramchandaniInes Kapferer-SeebacherEmine KandemisRasime KalkanPerla KalimanYoun Joung ChoBryan W. JonesFeras J. JirjeesEmilio JirilloChutima JirapinyoJunhyun JeonMusharraf JelaniAjit JaiswalMayumi IwasakiShinsuke ItoShumpei IshikawaRachel Helen HortonNele HoremansKi Ho HongGeorges HerbeinJohn E. HayesRuilian HanAgnieszka HalasKevin M. HaigisSanna HagmanZsuzsanna GurdanPramod Kumar GuptaA. GugliucciAlicja E. GrzegorzewskaDaniel GrimanelliStanislas Grassin-DelyleStephanie GraserIsabel Gon\u00e7alvesRicardo Santiago GomezJ. GohlkeShamila GinigeTapash Chandra GhoshRomain GherardiMeenu GhaiDavid P. GavinBrandon S. GautAmparo Garcia-TejedorPhilippe GallusciAlexandre GagnonCarlo GaetanoAttila FrigyesiJennifer L. FreemanMolly FoxNatalia ForgacovaAntonino ForaboscoEugenia Flores-AlfaroAlberto Fernandez-Jaenhongjuan FangLigia Carla Faccin-GalhardiWilliam EvansWon Sik EumEbru ErzurumluogluThomas EggermannDebasree DuttaBurak DurmazJorge Dom\u00ednguez-Andr\u00e9sKorcan DemirClelia De-la-Pe\u00f1aFrank de VriesMaria Corazon A. De UngriaDelcides Ferreira De Paula JuniorDaniel A. de LuisRaul de LucasJeremy J. DayAparup DasDr Amita CoutinhoRicardo D. ColettaAmander T. ClarkStephan ClaesThomas A. CiullaFabrizio CitarellaLuisa CimminoEr-Chieh ChoMaria Sole ChimentiZ. Jeffrey ChenZ. Jeffrey ChenPao-Yang ChenFuxue ChenPrashen ChelikaniPattama ChailertvanitkulCarlo CervellatiClarissa CataleValeria CarolaInmaculada Campos-GalindoBert CallewaertMatteo BusconiRussell J. BuonoSteven BuechlerRegina Maria Bringel MartinsJason H. BricknerJ. BrezinovaAlison C. BrewerAn BoudewynsYvonne B\u00f6ttcherG. BorgonovoPenelope E. BonnenLuca Reggiani BonettiDragana BogicevicPeyman Bj\u00f6rklundAndriy BilichakMaitree BhattacharyyaIwona Ben-SkowronekMoussa BenhamedAkeila Bellahc\u00e8neMaik BehrensMohamed A. BedaiwyDiane M. BecklesClaude BeckerDaniela Sanchez Bass\u00e8resManish BansalIstvan BaloghAlma BalestrazziAdayabalam S. BalajeeBahman BahramnejadMelinda R. BaerwaldFabian BaertlingAnwar BabanDiana A. Averill-BatesD. M. Avdjieva-TzavellaKapil K. AvasthiEliandro Reis TavaresEmine Ikbal Atl\u0131Naureen AslamS. AounzouGregory S. AntonarakisNorma Almaraz-AbarcaWatfa Al-MamariAhad AlizadehMohammad Yousef AlikhaniMercedes Alfonso-Prieto\u0130brahim AkalinAnnabel Ahuriri-DriscollGholamreza AhmadianWasim AhmadSarita AgarwalOladele Vincent AdeniyiRavinder AbrolFrancisco Abad-SantosNurul Syakima Ab Mutalib"} +{"text": "Following the publication of the original article , the autThe original article has beenAdditional file 3: Macro. 2step-TemplateMatching.ijm, available on the GitHub repository at https://github.com/multi-template-matching/MultiTemplateMatching-Fiji/blob/master/Fiji/scripts/Plugins/Template_Matching/2step-TemplateMatching.ijm. See Supplementary Material.pdf and Supplementary Figures.pdf for further information."} +{"text": "This article has been corrected: Due to errors during figure assembly, 10803-10815. https://doi.org/10.18632/oncotarget.2506Original article: Oncotarget. 2014; 5:10803\u201310815."} +{"text": "This article has been corrected: In 8162-8172. https://doi.org/10.18632/oncotarget.14131Original article: Oncotarget. 2017; 8:8162\u20138172."} +{"text": "This article has been corrected: In 1020-1030. https://doi.org/10.18632/oncotarget.2741Original article: Oncotarget. 2015; 6:1020\u20131030."} +{"text": "Correction to: Mol Cancer 9, 112 (2010)https://doi.org/10.1186/1476-4598-9-112Fig. Fig. Fig. Fig. Fig. Following publication of the original article , minor eThe corrected figure is given below. The correction does not have any effect on the results or conclusions of the paper. The original article has been corrected."} +{"text": "This article has been corrected: Due to errors during figure assembly, the fluorescence images in 6737-6748. https://doi.org/10.18632/oncotarget.3253Original article: Oncotarget. 2015; 6:6737\u20136748."} +{"text": "The authors would like to express their apologies and regret for the errors in the original published version of the abovementioned article. The corrected article follows.DOIshttps://doi.org/10.18332/tid/132833Original Article: https://doi.org/10.18332/tid/141989Correction: https://doi.org/10.18332/tid/142579Corrected Article:"} +{"text": "I would like to begin with a tremendous round of applause for all the authors, reviewers, and members of our editorial team who have continued to advance our knowledge of the microbial sciences during our second year of the COVID-19 pandemic.It\u2019s that time of year, as we head into the end-of-year holidays, to reflect on how the past 12\u2009months have played out at I alluded to the following in last year\u2019s message but would like to drive it home again, with the advantage of some hindsight. In early 2020, very few people would have predicted that by the end of the year we would have had an effective vaccine against SARS-CoV-2, much less multiple versions. Yet, we accomplished it, in no small part due to years of fundamental research on how the immune system recognizes and responds to pathogens, how mRNAs are synthesized and translated, and how to effectively deliver macromolecules into cells. Much of this progress was anchored in the microbial sciences: studies of viral immunology, landmark work on mRNA metabolism in bacteria and virally infected cells, understanding how pathogens bind to and enter cells, development of techniques that were originally used to introduce viral oncogenes into mammalian cells in culture, and the use of viral vectors for gene therapy all set the stage for the COVID-19 vaccines. I would be remiss if I failed to note that this research was performed all around the world.mSphere, we are committed to continuing to publish important studies that advance our knowledge of the plethora of microbes and how they interact with their hosts. I have no doubt that this research will pay enormous benefits in the future.At Our submission numbers remain high and, despite a small increase in our time to first decision late last year and early this year, we have turned that curve around and aspire to get our decision-making speed back to where it was in the \u201cBefore Times.\u201dI want to give special mention to the staff at ASM who have worked tirelessly to keep the gears of publication well oiled. In particular, Nikki Glenn, Amanda Donaldson, and Noel Lin have made our jobs as editors easier and delightful.ad hoc reviewers in 2021. They have our gratitude. I hope that you are able to enjoy some time over the next few weeks connecting or reconnecting with your families and loved ones, this year in person. Humans are social animals, and it\u2019s great to be able to resume these activities.Kjersti AagaardMohamed M. H. AbdelbarySabrina AbsalonMichael C. AbtMark D. AdamsJosephine Azikuru AfemaKayode Olayinka AfolabiSurya D. AggarwalHector Aguilar-CarrenoChristian Paul AhearnBrian M. M. AhmerMustafa AkkoyunluMd. Tauqeer AlamAshraf Al AshhabM. John AlbertAnoop AlexCaroline AlfieriHolly M. Scott AlgoodJonathan AllenEmma Allen-VercoeJuan C. AlonsoFrancis AlonzoChristopher AlteriJohn AlverdyChristopher S. AndersonMatthew Zack AndersonDavid R. AndesLaura Maria Andrade De OliveiraMarco AndreolliDiego O. AndreyAlberto AntonelliYoshiteru AoiCristian ApetreiChelsie Elizabeth ArmbrusterSandra K. ArmstrongJennifer M. AuchtungTatjana Av\u0161i\u010d-\u017dupancDomenico Azarnia TehranSophie Bachellier-BassiMichael A. BachmanSteffen BackertMatthew BaidemeCamden R. BairJonathon L. BakerKatherine H. BakerScott BalibanJimmy D. BallardGuilia BandiniFernando BaqueroNoa Barak-GavishJoseph T. BarbieriBrianne BarkerJason C. BartzMartine BassilanaChristine Marie BassisTilman BaumstarkMarco BecherelliSara BeierDaniel P. BeitingGeorgios N. BelibasakisAeriel D. BelkSamantha L. BellJessica A. BelserJorge L. BenachJose A. BengoecheaPeter BergholzTeresa M. BergholzTanja Beri\u0107David BernsteinStefan BertilssonRalph BertramSanchita BhadraDipankar BhattacharyyaBijit BhowmikFadil A. BidmosClaire H. BirkenheuerJacob P. BitounDaniel Blanco-MeloJon S. BlevinsJoseph M. BlissPatricia Pringle BloomAntje BlumenthalKasun H. BodawattaPierre BogaertsGregory BonitoS\u00e9bastien Bontemps-GalloAngela BordinJens BosseAnna BothTravis BourretKate BowermanEric BoydEthna Fidelma BoydTodd BradleyRita BrancoKyndall BraumullerLinda BreedenMathieu BrochetNichole A. BroderickChristopher B. BrookeGrayson BrownJeremy S. BrownKevin M. BrownMichael G. BrownHarry BrumerDonald A. BryantAlison BuchanLori L. BurrowsKaren BushAndrea CabibbeLaty A. CahoonYi CaiEloiza Helena CampanaEdgar I. Campos-MaduenoEric CaragataAlessandra CarattoliFranck Gael CarboneroMiguel Carda Di\u00e9guezJeffrey CareyRyan B. CarnegieJaime CarrascoVern B. CarruthersLeslie S. CaseyIrene CastanoSantiago Castillo-Ram\u00edrezClayton C. CaswellRodrigo Cay\u00f4Daniel CazaresBrandi N. CeliaNuno CercaMiguel Angel CevallosDipshikha ChakravorttyDouglas L. ChalkerThomas M. ChambersJosephine R. ChandlerMichael ChandlerRobert L. CharleboisSujata S. ChaudhariNeeraj ChauhanDamien ChaussabelMichael S. ChausseeFrancisco P. ChavezLiang ChenChiuping ChengRachel A. ChengLaurent Roberto ChiarelliAlex W. H. ChinMichaelle ChojnackiStephen A. ClarkErika C. ClaudDavid W. ClearySara ClohiseyShira Milo CochaviDarrell CockburnAshley CohenSean ConlanLaura CookGretchen CooleyBrendan CormackPierre CornelisCaitlin CossaboomSiobhan C. CowleyRobert A. CramerMax CravenerAlison K. CrissKarissa L. CrossRobert W. CrossLiwang CuiPaul J. CullenNatacha CuotoCameron R. CurrieTodd Andrew CuttsDennis G. CvitkovitchF. Heath DamronAjai A. DandekarStephen DanielsBiswadip DasBryan W. DaviesCharles R. DeanJean-Winoc DecousserElizabeth N. De GaspariMiranda De GraafKirk W. DeitschHarry P. De KoningFrank R. DeLeoThomas G. DenesDavid W. DenningRajendar DeoraCynthia Ann DerdeynSteven C. DerrickJigar V. DesaiLalitagauri DeshpandeSanjay Kumar DeyVijaykrishna DhanasekaranRishu DheerRobert P. DicksonDiego G. DielBeatriz Diez MorenoStephen P. DiggleJoseph P. DillardSiyuan DingMarc S. DionneAlan Angelo DispiritoDirk P. DittmerEunsoo DoCarlota Doba\u00f1o LazaroYohei DoiJanet DonaldsonCaihong DongMatthew J. DormanLaurent DortetBeno\u00eet DoubletCharles M. DozoisJan Felix DrexlerYuchun DuElves DuarteEdward G. DudleyBreck A. DuerkopAnne K. DunnSanjucta DuttaKathryn EatonLeo EberlKathryn M. EdenboroughTom EdlindElizabeth A. EdwardsMaren EggersSabine EhrtPatrick EichenbergerWaldir P. EliasJeremy R. EllermeierRoland EllingNajib M. El-SayedMostafa S. ElshahedJoanne B. EmersonVirve Irene EnneEeva Liisa Eronen-RasimusAlice L. ErwinJavier Antonio Escobar-PerezMatthew J. EvansFranziska FaberRobert FaganChristina S. FahertyLinda FalgenhauerS\u00e9amus FanningMauricio J. FarfanMatthew L. FaronAmy K. FeehanMario F. FeldmanJinrong FengJ. Christopher FennoDavid J. FergusonIsabel Fern\u00e1ndez EscapaAstrid FerrerRichard L. FerreroKenneth A. FieldsJoshua FiererSergio R. FilipeMaria F. FillatScott G. FillerDouglas K. FischerCarlos FloresStephanie FlowersFabrizio FoieniSteven L. FoleyLaura FordJarrod R. FortwendelMichael T. FranceKristi L. FrankNatalia FreundGeorg FritzInga Fr\u00f6dingTakasuke FukuharaMarta M. GagliaHannah GaimsterRaj GajiJames E. GalenMarkus GanterMichael G. GanzleErin C. GarciaSarahi L. GarciaAmy Shirley GargisKathleen G\u00e4rtnerCaroline Attardo GencoNoel GeraldCarmen GherasimLorenzo GiacaniHeather L. GlasgowOleg GlebovErin S. GloagMarek GniadkowskiRichard V. GoeringGustavo H. GoldmanJonathan Louis GolobBenjamin GolombLaura G\u00f3mez-ConsarnauAngela Gomez-SimmondsYanhai GongJesus Gonzalo-AsensioSteven D. GoodmanTobias GorisMorgan GorrisRia GoswamiMatthias GotteRevathi GovindManish GoyalAndreas GrabruckerLisa GralinskiLuke R. GreenAlexander L. 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HoweGongzheng HuKe HuLinden T. HuStephen S. H. HuangEili HuhtamoLewis HunJason F. HuntleyJillian H. HurstBonnie L. HurwitzWilhelmina HustonJustin HutchisonAlbina IbrayevaMelissa IngalaThomas J. InzanaWilliam W. JaMary Ann Jabra-RizkCody B. JacksonAnna C. JacobsWilliam R. JacobsGuilhem JanbonIngmar JanseMichael A. JarvisVicki JeffersNiuniu JiDong-Yan JinWilliam JohnsonSusan JosephLok R. JoshiYuan JunBarbara C. KahlMaria KalamvokiSuzanne R. KalbJeremy Phillip KamilManabu KannoFathi KarouiaAnbu Kumar KaruppannanFatah KashanchiJoseph KeaneDaniel B. KearnsScott P. KeelyEliisa Kek\u00e4l\u00e4inenBrendan KellyVolkhard A. J. KempfArnaud Kengmo TchoupaNemat O. KeyhaniShabaana A. KhaderArifa S. KhanM. Nadeem KhanNiharika KhannaKrystyn KiblerMegan R. KiedrowskiNicolas KiefferPeter E. KimaSamantha Jane KingDavid L. KirchmanThomas KislingerKimberly A. KlineMatthew D. KociAndrew Young KohTakahiko KoizumiJames B. KonopkaKaren A. KormuthDaniel KornitzerAnna D. 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MiuraTim MiyashiroKazufumi MochizukiMoniruzzaman MohammadIgor MokrousovChristopher MontgomeryCatherine MooreGonzalo MoratorioKaren MoreauHiroshi MoriKoji MoriKouichi MoritaMinoru MoriyamaThomas E. MorrisonJohn P. MorrisseyJulie MorrisseyKathryn MorrisseyJoachim Morschh\u00e4userKenichiro MotomuraW. Scott Moye-RowleyElke M\u00fchlbergerConrad W. MullineauxMatthew A. MulveyCarol A. MunroPeter J. MylerAnusha NaganathanMoon H. NahmRyosuke NakaiTeruaki NakatsujiSham NambulliVinay Kumar NandicooriCassandra E. NelsonUjjwal NeogiJeniel E. NettIrene L. G. NewtonRyan J. NewtonStuart T. NicholWright W. NicholsTracy L. NicholsonWilliam NicholsonPaula NiewoldAngela M. NiliusMatthew L. NillesYosuke NishimuraAngela Helen NobbsAna Rita NogueiraSteven J. NorrisJoshua D. NosanchukTakuro NunouraKlaus N\u00fcssleinAustin S. NuxollTorsten OchsenreiterNkechi Martina OdogwuKyle L. O'DonnellKazuhiro OgaiKnut OhlsenYusuke OkazakiFaten A. OkdaAndrew J. OliveAntonio OliverMartin OlivierMichael OlsonTeresa R. O'MearaAndres Opazo-CapurroRobert C. OrchardJohn Osei SekyereAndrei L. OstermanMartin PabstChristopher PaddockMalcolm G. P. PageJohn V. PaiettaGlen E. PalmerTimothy PalzkillJohn C. PanepintoMarcus PanningSneh Lata PanwarCostas C. PapagiannitsisDaniel Paredes-SabjaHyunsook ParkDane ParkerGabriel I. ParraColin R. ParrishSally R. PartridgeSamir N. PatelMark E. PeeplesJos\u00e9 PenadesXinxia PengDaniel PensingerMari PentJakob PernthalerAndreas PeschelKarin E. PetersonMichael PetridisMelinda M. PettigrewRenata Cristina Pic\u00e3oAndrzej PiekarowiczSeth H. PincusAmeet J. PintoJohann PitoutPaul J. PlanetMateusz PlucinskiMircea PodarShawn W. PolsonStephen J. PolyakVineel Kumar Pothi ReddyThomas PottageArjun PrasadVibhu PrasadGregory P. PriebeFirdausi QadriShangshang QinJianming QiuMaxime Qu\u00e9batteCheryl L. QuinnRobert Andrew QuinnBrent RaceGireesh RajashekaraStuart A. RalphSrinivasan RamakrishnanKumaran S. RamamurthiSasirekha RamaniMaria Soledad RamirezJoshua P. RamsayTara M. RandisLeslie Anne RankXiancai RaoChad A. RappleyePhilip N. RatherMaria Angelica ReaMichael D. ReedJose A. Regla-NavaJonathan S. ReichnerAaron ReinkeJyothi RengarajanNilton RennoPeter ReutherKelly C. RiceDave RichardAmariliz RiveraCarmen Alicia Rivera P\u00e9rezIan S. RobertsMarilyn C. RobertsD. Ashley RobinsonLibia Zulema Rodriguez AnayaFrancisco Rodriguez-ValeraAdriana E. RosatoDavid A. RosenHelene F. RosenbergIsabelle Rosinski-ChupinBenjamin RossRaymond R. RowlandClaudia R\u00fcckertThomas A. RussoJenna RychertDavid A. SackXavier SaelensChayan Kumar SahaPaula S. SalgadoFrancisco SalinasAnastasios SamarasJames E. SamuelOrjan SamuelsenDerrick SamuelsonJohn C. SamuelsonSarah E. SansomAna Carolina Carolina de Mello SantosManuela SantosChristian Santos-Medell\u00ednSunil D. SarojMichael Joseph SatlinYuya SatoTimothy J. SavageCharles A. ScangaConnie SchmaljohnThomas M. SchmidtMirco SchmolkeRaymond SchuchStacey Schultz-CherryWilliam R. SchwanJulia A. SchwartzmanAlison J. ScottIngrid L. ScullyAnna Maria SeekatzEinat SegevJulie A. SegreRyan F. SeipkeRanjan SenKarol SestakWilliam M. ShaferRebecca S. ShapiroNikki W. ShariatAmit SharmaAshu SharmaVijay K. SharmaThomas J. SharptonRoss ShawTimothy P. SheahanAlaullah SheikhSamuel A. ShelburneAimee ShenRobert C. ShieldsShin-Ru ShihTakaaki ShimohataNaoya ShinzatoAndrey N. ShkoporovNahum Y. ShpigelJoshua D. ShroutDavid SibleyLynn L. SilverMarie SimoninAnthony P. SinaiAmit SinghHerman SintimBeate M. SlabyLeyla SlamtiAmy SmithWiep Klaas SmitsEvan S. SnitkinFernando C. SonciniStephanie D. SongHarini SooryanarainDaniel O. SordelliJoseph A. SorgPaul SpearmanFabio M. SquinaJustina StanislawMaja StanojevicJack T. StapletonMichael N. StarnbachJacob L. SteenwykStefania StefaniWilliam J. SteinbachSilke StertzDavid Cole StevensMarc J. A. StevensGeorge C. StewartAlison Elizabeth StoutDaniel StraumeJorg StulkeDerek SullivanWei SunYingjie SunJoyce SutcliffeTroy C. SuttonYasuhiro SuzukiMary Hannah SwaneyToru TakeshitaToru TakimotoMichal Caspi TalSatoshi TamazawaMichelle D. TateTallita C. L. TavaresVeronika TchesnokovaRobin TeconSam R. TelfordBen TempertonFred C. TenoverAkihiko TeradaAartjan J. W. te VelthuisRajagowthamee ThangavelKevin R. TheisJustin A. ThorntonYun TianCaroline T. TiemessenAnna D. TischlerKelvin K. W. ToRichard B. ToddMasanori TohnoHirokazu TojuMark Alexander TolemanAndrew TomarasJavier TorresDieter M. TourlousseMaria Carolina TouzJulian TrachselAndrej TraunerPankaj TrivediBilly TsaiAthanassios TsakrisBoo Shan TsengClaire Elizabeth TurnerJane F. TurtonJessie UehlingJuan Esteban UgaldeGottfried UndenSyun-Ichi UrayamaConstantin F. UrbanBlake UshijimaDavid W. UsseryPeter Valentin-WeigandMark van der GiezenPeter Van der VoortNicole N. van der WelPatrick Van DijckGiel G. van DoorenDavid van DuinDaria Van TyneArvind VarsaniJos\u00e9 Antonio V\u00e1zquez-BolandKasthuri VenkateswaranElisa M. VeselyJorge E. VidalRafael VignoliMarius VitalJay VornhagenMartin I. VoskuilDaniel E. VothSlavena VylkovaDavid M. WagnerSeth T. WalkEdward WalkerKimberly A. WalkerMark J. WalkerGemma WaltonChengming WangKai WangShanquan WangTony WangXiaoxue WangYang WangDavid Morgan WardEmma WearMary WeberNa WeiTiffany WeinkopffBrian C. WeinrickBrian WeissTao WenGuido WernerLukas WeseslindtnerDave J. WestenbergEdze WestraDawn WetzelNathan John WeyandRobert T. WheelerJudith M. WhiteShannon WhitmerGregory WiedmanKrista Rule WiggintonMichael R. WileyBrian J. WilkinsonJulia WillettSimon WilliamsMary E. WilsonJeffrey H. WitheyGeorge WittemyerChristiane E. WobusBrian Allan WolffAdam C. N. WongThomas WoodStephen M. WoodcockMichael H. WoodworthAniela WozniakRachel A. F. WozniakXianfu WuLihua XiaoHang XieChenggang XuJianping XuJin-Rong XuKageto YamadaKyosuke YamamotoShinji YamasakiS. Steve YanTao YanHee-Jeong YangZhaomin YangHui-Ling YenJae-Hyuk YuYunsong YuJoseph P. ZackularRaffaele ZarrilliEgija ZauraAhmed ZayedGuoquan ZhangLianhui ZhangRui ZhangWanjiang ZhangWen ZhangXilin ZhaoYaofeng ZhaoBeiwen ZhengYong ZhengXiaohui ZhouDuolong ZhuGuoqiang ZhuJoseph M. ZiegelbauerStefan ZimmermannFinally, we list below the many individuals who served as"} +{"text": "This article has been corrected: Due to errors during figure assembly, an accidental duplicate image of panel 4 in 4814-4825. https://doi.org/10.18632/oncotarget.13978Original article: Oncotarget. 2017; 8:4814\u20134825."} +{"text": "LeukemiaCorrection to: 10.1038/leu.2008.89The authors have retracted this article . An inve"} +{"text": "This article has been corrected: The image in 66922-66934. https://doi.org/10.18632/oncotarget.11877Original article: Oncotarget. 2016; 7:66922\u201366934."} +{"text": "Cerebral organoids are three-dimensional cell-culture systems that represent a unique experimentalmodel reconstructing early events of human neurogenesis in vitro in health and various pathologies. The mostcommonly used approach to studying the morphological parameters of organoids is immunohistochemicalanalysis; therefore, the three-dimensional cytoarchitecture of organoids, such as neural networks or asymmetricinternal organization, is difficult to reconstruct using routine approaches. Immunohistochemical analysis of biologicalobjectsis a universal method in biological research. One of the key stages of this method is the productionof cryo- or paraffin serial sections of samples, which is a very laborious and time-consuming process. In addition,slices representonly a tiny part of the object under study; three-dimensional reconstruction from the obtained serialimages is an extremely complex process and often requires expensive special programs for image processing.Unfortunately, staining and microscopic examination of samples are difficult due to their low permeability and ahigh level of autofluorescence. Tissue cleaning technologies combined with Light-Sheet microscopy allows thesechallenges to be overcome. CLARITY is one of the tissue preparation techniques that makes it possible to obtainopaque biological objects transparent while maintaining the integrity of their internal structures. This method isbased on a special sample preparation, during which lipids are removed from cells and replaced with hydrogelcompounds such as acrylamide, while proteins and nucleic acids remain intact. CLARITY provides researchers witha unique opportunity to study three-dimensional biological structures while preserving their internal organization,including whole animals or embryos, individual organs and artificially grown organoids, in particular cerebralorganoids. This protocol summarizes an optimization of CLARITY conditions for human brain organoids and thepreparation of Light-Sheet microscopy samples. Biological tissues and organs present a complex three-dimensionalstructure. Due to their opacity and high level ofautofluorescence, three-dimensional reconstruction of suchobjects is an extremely laborious, but necessary process.To date, a number of protocols (more than a dozen) havebeen developed for making tissue transparent: SeeDB , ScaleA2 , uDISCO , CLARITY , CUBIC and others. In general, all protocols canbe divided into three groups, depending on the chemicalsused for tissue clearance: organic solvents (hydrophobicreagent)-based protocols ,hydrophilic reagent-based protocols and hydrogel-tissuechemistry-based protocol . Some of themhave different advantages like quality and speed of clearingor simplicity of the procedure. But on the other hand, some ofthe protocols involve using toxic and corrosive chemicals thatrequire special objectives to avoid damage to the microscopeor require other special equipment. Most of these protocolshave been developed to clarify entire organs or their bigfragmentsRecently, a new method of artificial mini-organ or organoidsgeneration from induced pluripotent stem cells (iPSC) wasdeveloped and now many differenttypes of organoids have already been produced . Organoids arewidely used both to recreate the three-dimensional architectureand functional activity of the original organs during normalembryonic development and at various disorders and to test thebiological activity of various drugs, chemical and biologicalagents. Usually, organoids are opaque, which makes investigatingthem rather difficult. For this purpose, it is advisableto use the combination of tissue clearing and 3D imagingtechnologies. However, it is important to select the clarifyingtechnology that would match organoids size and fragility asmuch as possible and would produce sufficient resolution forinvestigation of tiny structures.Various techniques have been used for organoid tissueclearing and several studies have compared different clarifyingmethods which could be applied to mini-organs . Some techniques, such as the hydrophilicclearing protocols are most acceptablefor clearing small spheroids such as neurospheres or cancer cell spheroids . Others, such as RapiClear, Fructoseglycerol andFUnGI, also using hydrophilic components, are designed andoptimized for handling small and fragile, predominantly holloworganoid structures such as intestinal organoids. It shouldbe noted that these protocols are very convenient and take onlythree days without application of harmful chemicals . For complex and densebrain organoids, stronger clearing protocols including delipidationprocedure are usually used .Applying organic solvent-based methods like 2Eci (2nd generationEthyl cinnamate-based clearing method) or BABB methodfor midbrain organoids can get a relativelyquick (within a few days) result. However, most of theorganic components used in these protocols are quite toxic ).The use of hydrogel-tissue chemistry sometimes providesmore opportunities for preserving the structure of organoidsand increasing the optical resolution of tiny objects. That isdue to the tissue hydrogel scaffold preparation by cross-linkinghydrogel monomers to native biomolecules . The creation of such a polymer frame in the brainorganoids allows combining these protocols with additionalprocedures with sodium dodecyl sulfate and physical electrophoresis,as well as with high-resolution imaging of ExpansionMicroscopy with a general microscopy setup .Thus, it is quite important to choose the most optimal andeffective tissue clearing technique for samples, especially forsuch complex objects as cerebral organoids.One of the most convenient and lab-friendly techniques isCLARITY (Clear Lipid-exchanged Acrylamide-hybridizedRigid Imaging/Immunostaining/In situ hybridization-compatibleTissue-hYdrogel). CLARITY was developed in 2013for obtaining high-resolution information from complex3D structures, such as the whole mouse brain . Application of this technique enabled to obtainintact-tissue imaging of long-range projections, local circuitwiring, cellular relationships, subcellular structures, proteincomplexes, and neurotransmitters. CLARITY protocol includesreplacing lipids with hydrophilic polymers (acrylamideand bis-acrylamide), which help to stabilize tissue but make itoptically transparent and permeable. It is very important thatmolecules like nucleic acids and proteins stuck in the hydrogelkeep their structures and locations. Thus, CLARITY allowscombining tissue clearing techniques with immunostainingand in situ hybridization and explores the internal structureof large three-dimensional objects without damaging theirintegrity. There is only one article in which CLARITY techniquewas used for cerebral organoid clarifying , but without a detailed description. Thus, theaim of our work was optimization of CLARITY protocol inapplication to cerebral organoids and detailed description ofsamples preparation for Light-Sheet microscopy.Reagents1. Acrylamide .2. Agarose D1, low EEO .3. Bisacrylamide .4. Boric acid .5. ddH2O.6. Glue .7. Parafilm M .8. Paraformaldehyde (PFA) .9. Phosphate buffer saline (PBS) .10. Sodium azide .11. Sodium dodecyl sulfate .12. Triton X-100 .13. VA044 .14. Serological pipets 5, 10, 25 ml .15. 1-ml syringe .16. 2 ml tube .17. 5 ml tube .18. Glass bottle 100 and 500 ml .19. Syringe filter, 0.22 \u03bcm .20. 4\u2032,6-diamidino-2-phenylindole (DAPI) .21. Antibodies (Table 1).Equipment1. Light-Sheet Z1 microscope (Zeiss).2. Orbital shaker .3. Roller shaker .4. pH meter .5. Magnetic stirrer .6. Standard microwave.7. Thermometer.8. Forceps.9. Chemical spoons.10. Fume hood.11. Icebox.Software1. ImageJ 2. ZEN ProcedureFixation of human cerebral organoidNote: For any manipulation with organoids, use cut 1 ml tipsor wide orifice 1 ml tips to protect samples from damage.1. Transfer cerebral organoids in 5-ml tubes and wash with1X PBS solution 2 times.2. Replace 1X PBS solution with freshly prepared 4 % PFAsolution.3. Place the tubes on the roller/orbital shaker and incubate atroom temperature for 2\u20133 h.4. Wash samples 3 times with 1X PBS solution for 30 min.Note: At this step, cerebral organoids can be kept at +4 \u00b0C in1X PBS solution. For keeping more than 1 week, we recommendadding sodium azide to a final concentration of 0.01 %to prevent sample contamination with bacteria and fungi.Hydrogel embedding1. Precool all solutions, equipment, and samples on ice toprevent premature polymerization of the hydrogel solution.Note: If you use a frozen aliquot of hydrogel solutions, thawthe vial on ice in a fridge overnight. After thawing, gently mixand check for the absence of precipitation.2. Fill the 2-ml tube with the hydrogel solution and transfercerebral organoids in the tube having previously gentlyremoved leftovers of the PBS with a paper towel.Note: 2-ml tube format is acceptable for 1\u20133 organoids. Fora large number of organoids, we recommend using a biggertube.3. Incubate the samples in the hydrogel solution at +4 \u00b0C atthe lowest speed of roller/orbital shaker for 24 h.4. Refill the tube with fresh hydrogel solution and incubateat +37 \u00b0C for 4 h.Note: Fill the tube with hydrogel solution completely. Oxygeninhibits hydrogel polymerization, thus all bubbles should beremoved. Additionally, we recommend covering the tube withParafilm to prevent air access.5. Very gently extract the samples from the polymerizedhydrogel by carefully rolling samples on a paper towel.6. Transfer the samples into the 5-ml tube and wash withClearing Solution 4 times at room temperature for 24 h.Passive clearing1. Change Clearing Solution every 2 days and incubate at+37 \u00b0C with agitation. Continue clearing until samplesbecome transparent.Note: We strongly recommend using +37 \u00b0C for lipid removal.Room temperature slows this process down to several months!Note: The time of tissue clearing depends on the size of organoids.Cerebral organoids \u22640.5 cm become transparent during~2 weeks, for organoids \u22650.5 cm it can take up to 3 weeks.2. Wash samples in PBST for 48 h. Change solution 2\u20133 timesper day.Staining1. Incubate samples with primary antibodies in PBST at roomtemperature on a shaker for 3 days.2. Wash samples with PBST for 2 days, changing PBSTevery 4 h.3. Incubate with secondary antibodies and DAPI at roomtemperature for 2 days.4. Wash samples with PBSR for 2 days, changing PBSTevery 4 h.Note: Antibody consumption for staining of CLARITY samplesis very high. We recommend reducing the volume to the minimumat which the samples in the tube are completely coveredwith the staining buffer with constant stirring on an orbitalor roller shaker.Note: For larger organoids, we recommend extending eachstaining step by at least 1 daySample preparation for Light-Sheet microscopyOrganoid sizes can vary greatly. Therefore, we recommendusing a different fixation method for Light-Sheet microscopydepending on the size.Agarose embedding samples Note: Use the agarose with a low melting point temperatureonly.Note: The percentage of agarose solutions depends on thesize of the organoid. For larger organoids, use 1.5 % agarosesolution.1. Prepare the 1-ml syringe by cutting off the top .2. Weigh the required amount of agarose (at the rate of 1 g per100 ml) and dissolve in 1X PBS or ddH2O. Prepare agarosesolution by melting in the microwave. Usually, for a 1-mlsyringe, 1.5 ml of agarose is enough.3. Pour the hot agarose solution into a 12-well plate or anyother laboratory glassware or plasticware. When agarosesolution cools down to +40 \u00b0C, transfer samples and gentlymix. Put the samples in agarose solution into the 1-mlsyringe.4. Assemble the 1-ml syringe with a sample holder .5. Proceed to Light-Sheet microscopy.Note: Fill the microscope chamber with ddH2O or 1X PBS.No great differences were observed between the two solutions.Agarose-free or hanging samples (for bigger samples)1. Glue the sample to the bar . The area ofadhesion can be increased by attaching a small piece of filterpaper. Keep samples in ddH2O or 1X PBS before placingthem into the microscope chamber.2. Proceed to Light-Sheet microscopy.Note: It is imperative to check and rinse the rod and sampleholder for glue residues. If there are any, we strongly recommendthat you soak in soapy water and mechanically removeany glue residue.RecipesNote: Most solutions and reagents from this protocol are toxicand biohazardous. Do not forget about your safety and workin protective laboratory clothing and only under a fume hood!10X PBS solutionTo prepare a 10X stock solution, dissolve 10 tablets of PBSin 100 ml of ddH2O.PFA solutions\u2022 16 % PFA stock solutionTo prepare stock solution, dissolve 16 g of PFA in 80 mlof 1X PBS using a magnetic stirrer. Adjust pH to 7.4\u20137.5and add 1X PBS up to 100 ml. Filter the solution througha 0.40 \u03bcm filter and aliquote into 5 ml tubes. Keep stocksolution at +4 \u00b0C for short storage (up to 2 weeks) or at\u201320 \u00b0C for long storage.\u2022 4 % PFA working solutionTo prepare 4 % PFA working solution, dilute stock solutionwith 1X PBS.Hydrogel solutionNote: All solutions and equipment have to be pre-cooledto prevent premature polymerization of hydrogel solution.1. Mix all components on ice according to Table 2.2. Aliquote hydrogel solution and keep at \u201320 \u00b0C for longstorage or use freshly prepared solution.Clearing solution1. Mix all components on ice according to Table 3.2. Keep the solution in a glass bottle at room temperature.PBST1. Add Triton-X100 to the final concentration of 0.1 % usinga magnetic stirrer.2. Keep the solution in a glass bottle at room temperature.Human cerebral organoids were generated according to a protocolfrom Lancaster et al. (2013) with small modifications.2- and 3-month-old cerebral organoids were used for tissueclearing protocol. At this stage, there are dense spheres morethan 2 mm in diameter . We noted that the time oftissue clearing depends on the size of organoids. Cerebralorganoids \u2264 0.5 cm become transparent during ~2 weeks,for organoids \u2265 0.5 cm up to 3 weeks. This time may varyfrom sample to sample, however, continue cleaning until thesamples become transparent .For immunostaining we chose two proteins with differentsubcellular localisation such as nuclear CTIP2 andcytoplasmic bTubb3 . We did not find a significantdifference between penetration of antibodies into different cellular compartments. In both cases, we observed specificstaining throughout the entire thickness of the organoid .Cerebral organoids are a unique novel technology that allowsthe reconstruction of early human neurogenesis. Theoutstanding feature of this in vitro system is the reproductionof the three-dimensional organization of the human embryonicbrain. Standard histological methods of analysis do notallow reconstructing the internal structure of the cerebralorganoids and result in information loss. The tissue clearingtechnique helps to overcome these limitations, allowing torecreate a three-dimensional model of cerebral organoidsand explore their fine organization without internal structuredestruction. This is especially important for the investigationof brain organoids since they contain a dense network of longprocesses of nerve cells, which is very difficult to study byserial sections .Based on the various tissue clearance techniques analysis,we settled on the use of hydrogel-tissue chemistry as a clearingagent. Generally hydrophobic and hydrophilic reagent-basedprotocols are applied to the investigation of spheroids or holloworganoids such as intestinal organoids , while hydrogel reagents are used for the clarifying ofhuman iPSC-derived retinal organoids and iPSC-derived cerebral organoids . Hydrogel-tissuechemistry-based protocols maximize the preservation of theinternal structure of organoids and allow to achieve high opticalresolution and low background at fluorescent microscopyCurrently, there are at least three known hydrogel-tissuechemistry-based methods that use different delipidation anddehydration chemicals: SWITCH Diatrizoic acid N-methyl-D-glucamineIodixanol (dehydration)), SHIELD (Polyepoxy cross-linking(Delipidation), Diatrizoic acid N-methyl-D-glucamine Iodixanol(dehydration)) and CLARITY (Hydrogel embedding(Delipidation), HistodenzTM Glycerol (dehydration)) . Therefore in our choice ofa suitable technique, we also focused on the availability ofthe appropriate reagents, the simplicity of the protocol andthe lack of need for special equipmentOf course, a significant disadvantage of CLARITY techniqueis the relatively long tissue clearance procedure , butthis obstacle is compensated by a quite simple protocol. 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PLoS ONE 16(3): e0247913."} +{"text": "In the original article published online one of the references is incorrect, which seems to be preventing it from being indexed.It reads:https://doi.org/10.1016/j.futures.2021.102756.Jacy Reese, Anthis Eze, Paez (2021) Moral circle expansion: A promising strategy to impact the far future. Futures 130102756\u2013But it should read:https://doi.org/10.1016/j.futures.2021.102756Anthis, J.R., & Paez, E. (2021). Moral circle expansion: A promising strategy to impact the far future. Futures 130102756. The original article has been corrected."} +{"text": "This article has been corrected: In 6870-6878. https://doi.org/10.18632/oncotarget.27326Original article: Oncotarget. 2019; 10:6870\u20136878."} +{"text": "Correction to: Trials 17:809 (2021).https://doi.org/10.1186/s13063-021-05761-0Following the publication of the original article , we wereOriginally published name: Anne Tsay.Corrected name: Annie Tsay.The original article has been corrected."} +{"text": "Correction to: Oncogene;10.1038/onc.2015.355; Published online 21 Sept 2015In Fig."} +{"text": "Scientific Reports 10.1038/s41598-020-72133-0, published online 14 September 2020Correction to: The original version of this Article contained an error in the Acknowledgments section.\u201cThis work was funded by Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia through projects UID/QUI/00100/2019 and UIDB/00100/2020 to Centro de Qu\u00edmica Estrutural, through research grants SAICTPAC/0019/2015 and PTDC/QUI-QAN/32242/2017, and through PhD grants SFRH/BD/110945/2015 (P.F.P.) and SFRH/BD/143128/2019 (C.F.M.). K.S.B. thanks EMBO for providing the short-term fellowship (EMBO ref. 8107). M.M.A. and G.C.J. are IST-ID employees under contracts IST-ID/154/2018 and IST-ID/090/2018, respectively, in agreement with D.L. 57/2017. The authors would also like to thank Ana Charas for providing the ITO/PET substrate.\u201dnow reads:\u201cThis work was funded by Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia through projects UID/QUI/00100/2019 and UIDB/00100/2020 to Centro de Qu\u00edmica Estrutural, through research grants LISBOA-01-0145-FEDER-016405 \u2013 SAICTPAC/0019/2015 and PTDC/QUI-QAN/32242/2017, and through PhD grants SFRH/BD/110945/2015 (P.F.P.) and SFRH/BD/143128/2019 (C.F.M.). K.S.B. thanks EMBO for providing the short-term fellowship (EMBO ref. 8107). M.M.A. and G.C.J. are IST-ID employees under contracts IST-ID/154/2018 and IST-ID/090/2018, respectively, in agreement with D.L. 57/2017. The authors would also like to thank Ana Charas for providing the ITO/PET substrate.\u201dThe original Article has been corrected."} +{"text": "Scientific Reportshttps://doi.org/10.1038/srep27911, published online 15 June 2016Correction to: This Article contains errors.For Figure"} +{"text": "The original version of this article unfortunately contained a mistake. The following author names were incorrectly structured.Tobias C. WoodGareth J. BarkerJuan A. Hernandez-TamamesEsther A. H. WarnertThe original article has been corrected."} +{"text": "Scientific Reports 10.1038/s41598-020-75298-w, published online 27 October 2020Correction to: This Article contains an error. A Data Availability section was originally not included\u2014it should appear as below:Data AvailabilityData underlying this study can be accessed through the Cranfield University repository at 10.17862/cranfield.rd.13186745.v1."} +{"text": "Emergomyces orientalis Emergomycosis Diagnosed by Metagenomic Next-Generation Sequencing . The article has been corrected online (https://wwwnc.cdc.gov/eid/article/27/10/21-0769_article).The name of author Xiaohui Wang was misspelled in"} +{"text": "This article has been corrected: In 39907-39915. https://doi.org/10.18632/oncotarget.9530Original article: Oncotarget. 2016; 7:39907\u201339915."} +{"text": "Correction to: BMC Palliat Care 21, 13 (2022)https://doi.org/10.1186/s12904-021-00881-5Following the publication of the original article , the lasJony Francisco Dos Santos Silva. Identifikasi Kebutuhan Palliative Care pada Pasien Penyakit Kronis di Ruang Rawat Inap Dewasa di RSUP Dr. Sardjito Yogyakarta. Thesis. 2020. Yogyakarta. UnpublishedThe original article has been updated."} +{"text": "This article has been corrected: Due to accidental placement, some of the images in 86488-86502. https://doi.org/10.18632/oncotarget.21212Original article: Oncotarget. 2017; 8:86488\u201386502."} +{"text": "J Clin Invest. 2007;117(11):3211\u20133223. https://doi.org/10.1172/JCI31757Original citation: J Clin Invest. 2022;132(1):e157373. https://doi.org/10.1172/JCI157373Citation for this corrigendum: The authors recently became aware that the image for the HA (Foxo3a) blot in The authors regret the error."} +{"text": "This article has been corrected: In 16531-16552. https://doi.org/10.18632/oncotarget.14972Original article: Oncotarget. 2017; 8:16531\u201316552."} +{"text": "This article has been corrected: The 23018-23028. https://doi.org/10.18632/oncotarget.25195Original article: Oncotarget. 2018; 9:23018\u201323028."} +{"text": "Open Biol.12, 210206. (Published online 16 February 2022). (doi:10.1098/rsob.210206)The originally published version of this paper showed the incorrect figure legends for figures"} +{"text": "JCI Insight. 2016;1(17):e87754. https://doi.org/10.1172/jci.insight.87754Original citation: JCI Insight. 2021;6(7):e149896. https://doi.org/10.1172/jci.insight.149896Citation for this corrigendum: The Editors previously posted an Expression of Concern for this article regarding images in Supplemental 4B that appeared similar (Endo40 and Endo42 samples) and images in The authors regret the error."} +{"text": "Correction to: BMC Pediatr 21, 311 (2021)https://doi.org/10.1186/s12887-021-02517-2After publication of this supplement article , it was The references have since been corrected in the original article and may be found detailed below:https://www.nice.org.uk/guidance/ph31[35] NICE. Unintentional injuries on the road: interventions for under 15s [Internet]. 2010 [cited 2019 Oct 21]. p. 1\u201346. Available from: https://www.nice.org.uk/guidance/ph29[43] NICE. Unintentional injuries: prevention strategies for under 15s [Internet]. 2010 [cited 2019 Oct 21]. p. 1\u201388. Available from: [67] Watson M, Kendrick D, Coupland C, Woods A, Futers D, Robinson J. Providing child safety equipment to prevent injuries: randomised controlled trial. BMJ. 2005;330:178\u201382.The author apologizes for any inconvenience caused."} +{"text": "Neovison vison), Poland . The article has been corrected online (https://wwwnc.cdc.gov/eid/article/27/9/21-0286_article).The acknowledgments and funding information were inaccurate in Severe Acute Respiratory Syndrome Coronavirus 2 in Farmed Mink ("} +{"text": "This article has been corrected: In 2546-2560. https://doi.org/10.18632/oncotarget.26817Original article: Oncotarget. 2019; 10:2546\u20132560."} +{"text": "Diptera: Scenopinidae), Scenopinusjereisp. nov., with characteristic bicoloured legs and completely black halteres, is described from Finland. To exclude potential previously named species, a survey of the relevant type specimens as well as original descriptions of the Palearctic and Nearctic Scenopinus species has been conducted, including old Scenopinusfenestralis (Linnaeus) synonyms. Scenopinusjereisp. nov. is likely to be an overlooked, boreal forest specialist living in the nests of cavity-nesting birds. An identification key to the European species is provided.A new species of window fly ( Diptera: Scenopinidae) are a small family of primitive flies belonging to the therevoid clade of the Asiloidea superfamily . The family has a cosmopolitan distribution, with more than 420 described species in 25 genera. Scenopinidae consists of three subfamilies, Caenotinae (1 genus), Proratinae (6 genera), and Scenopininae (18 genera) , S.bouvieri , S.glabrifrons Meigen, 1824, S.gobiensis Kelsey, 1981, S.griseus , S.halteralis Frey, 1936, S.lesinensis Strobl, 1902, S.niger , S.oldenbergi (Kr\u00f6ber) , S.verrucosus Carles-Tolra, 2001, and S.vitripennis Meigen, 1824. In addition, S.phaidimos Kelsey, 1969 is present in Turkey and might be expected to occur in the eastern Mediterranean.Window flies ( genera) , with on genera) : CaenoneScenopinidae is in the arid regions of the world directly to vials and killed by freezing, ethyl acetate or potassium cyanide prior to mounting them on entomological pins.Apart for the two old museum specimens of Label data of newly collected specimens are given verbatim using the following symbols: / end of a line and beginning of the next; // end of label and beginning of the next (from top to bottom on the same pin). The specimens are deposited in the following collections and are indicated with the given acronym in the text:AHC Private collection of Antti Haarto, Myn\u00e4m\u00e4ki, FinlandJPC Private collection of Jaakko Pohjoism\u00e4ki, Joensuu, FinlandMIZMuseum and Institute of Zoology, Polish Academy of Sciences, Warszawa, PolandMZHFinnish Museum of Natural History, Zoological Museum, University of Helsinki, Helsinki, FinlandMZTZoological Museum of the University of Turku, Turku, FinlandSMNSStaatliches Museum f\u00fcr Naturkunde Stuttgart, Germanyhttps://laji.fi/theme/emk for more details) and coordinates (NNNN:EEEE) on the labels are mostly given in the old national Finnish map grid coordinate system 1\u2642(dissected): Fennia Kb [Karelia borealis \u2013 north Karelia]: Liperi/ Kontkala 6950:3616 / 19.7.2014/ Ali Karhu leg. [JPC]; 1\u2640: Same collection data ; 1\u2640: Finland, Kb: Ilomantsi/ Kelovaara 70008:36825 / 25.6.2016/ J. Pohjoism\u00e4ki leg. [JPC]; 1\u2640: Finland, Sa [Savonia australis \u2013 south Savo]: Taipalsaari/ Riihilahti 6778:3564 / 21.7.2015/ J. Pohjoism\u00e4ki leg. ; 1\u2642: Finland, Ta [Tavastia australis \u2013 south H\u00e4me]: Orivesi/ Siitama 6835:3354 / 11.7.2009/ J. Pohjoism\u00e4ki leg. [JPC]; 1\u2640: Finland, Ok [Ostrobottnia kajanensis \u2013 Kainuu region]: Sotkamo/ Laukkala, 7114:3565 / 1.7.2005/ J. Pohjoism\u00e4ki leg. [JPC]; 1\u2640: Finland, Ab [regio Aboensis \u2013 Turku region]: Myn\u00e4m\u00e4ki/ Perkko 6733:3222 / 22.7.2011/ A. Haarto leg. . 1\u2642: Finland, Ab: Myn\u00e4m\u00e4ki/ Perkko 6733:3222 / 13.6.2009/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ fenestralis (L.)/ det. A. Haarto 2009/ AHa09\u2013000593 [MZT]; 1\u2640: Finland, Ab: Mietoinen/ Perkko 6733:[3]222 / 17.7.2003/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ fenestralis (L.)/ det. A. Haarto 2008/ AHa08\u2013001324 [AHC]; 1\u2640: Finland, Ab: Mietoinen/ Perkko 6733:[3]222 / 17.7.2003/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ fenestralis (L.)/ det. A. Haarto 2008/ AHa08\u2013001324 [AHC]; 1\u2642: Finland, Ab: Mietoinen/ Perkko 6733: [3]222 / / 16.5.2004/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ fenestralis (L.)/ det. A. Haarto 2021/ AHa21\u2013000589 [AHC]; 1\u2642: Finland, Ab: Mietoinen/ Perkko 6733: [3]222 / 5.6.2004/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ fenestralis (L.)/ det. A. Haarto 2021/ AHa21\u2013000590 [MZT]; 1\u2642: Finland, EP [Etel\u00e4-Pohjanmaa]: Isokyr\u00f6/ Orisberg 6983: [3]265 / 7.7.1999/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ fenestralis (L.)/ det. A. Haarto 1999 [AHC]; 1\u2640: Same collection and determination data [AHC];Germany: 2\u2640\u2640: Germany/ Hessen, Friedberg/ Ockstadt 50.3319, 8.7208 [Geographic coordinate]/ 13.6.2010/ J. Pohjoism\u00e4ki leg. [JPC]Greece: 2\u2640\u2640: GR CRETE Chania/ Thymia 35.4106, 24.0440 [Geographic coordinate]/ 5.-6.vi.2019/ J. Pohjoism\u00e4ki leg. [JPC]Finland: 6\u2642\u2642, 4\u2640\u2640. See the type material below for details.Finland: 2\u2640\u2640: Finland, Sa: Kouvola, 674\u2013679:347\u2013350 / e.l. 2018 ex Strixaluco nest box. / M. Mutanen leg. [JPC]; 1\u2642: Finland, Sa: Taipalsaari/ Riihilahti 6778:3564 / 21.7.2015/ J. Pohjoism\u00e4ki leg. ; 1\u2640: Finland, Ta: Tampere/ Rantaperki\u00f6 6822:3327 / 26.6.2009/ J. Pohjoism\u00e4ki leg. [JPC]; 1\u2640: Finland, Ab: Myn\u00e4m\u00e4ki/ Perkko 6733:3222 / 12.6.2011/ A. Haarto leg. ; 1\u2640: Finland, V [Varsinais-Suomi]: Turku Hirvensalo/ Rauhala 6707:[3]233/ 22.5.1996/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ niger (DeGeer)/ det. A. Haarto [AHC];] ; 1\u2642: Finland, V: Turku Hirvensalo/ Rauhala 6707: [3]233 / 7.6.1996/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ niger (DeGeer)/ det. A. Haarto [AHC]; 1\u26421\u2640: Finland, V: Turku Hirvensalo/ Rauhala 6707: [3]233 / 7.6.1996/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ niger (DeGeer)/ det. A. Haarto [MZT]; 1\u2642: Finland, Ab: Myn\u00e4m\u00e4ki/ Perkko 67333:32223 / 22.5.2017/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ niger (De Geer)/ det. A. Haarto 2017/ AHa17\u2013001063 [AHC]; 1\u2640: Finland, V: Kaarina/ Kuusisto R\u00f6varholm/ 9.7.1998/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ niger (De Geer)/ det. A. Haarto 2008/ AHa08\u2013001326 [AHC].Germany: 1\u2642: Scen. glabrifrons/ W\u00fcrttbg Meig. ?/ v.Roser 1872 [handwritten]// Scenopinus/ vitripennis Meig./ det. L.P. Kelsey 1964 [SMNS]. Examined from high resolution photographs. See the discussion regarding the identity of this specimen.Poland: 1\u2640: Warszawa [barely visible]/ 14.vii.1953 r./ leg. R. Trojan// Omphrale \u2640/ vitripennis (Meig)/ P. Trojan det. 1954. [MIZ]. Examined from high resolution photographs.The classification follows Herting and Dely-Draskovits (1993). The morphological terminology used in this study follows The images were taken with a Leica Z6APO stereomicroscope and a Leica DFC450c (5MPix) camera, MSV266 motorised focus and using the Leica Application Suite 4.6.0 software for Z-axis stacking. Images were cropped, colour- and contrast-enhanced but not manipulated otherwise.COI) DNA barcoding was performed as a part of the Tachinidae project of Finnish Barcode of Life initiative (FinBoL). The 5\u00b4-terminal part of COI was amplified using the routine barcoding primers LepF1 and LepR1 (BOLD) are given for each barcoded specimen.Cytochrome oxidase subunit 1 : Finland, Sa: Kouvola, 674\u2013679:347\u2013350 / e.l. 2018 ex Strixaluco nest box. / M. Mutanen leg. // Scenopinusjerei sp. nov. Pohjoism\u00e4ki & Haarto 2021 / (Diptera: Scenopinidae) / J. Pohjoism\u00e4ki det. // HOLOTYPE [red label] [MZH] Paratypes: 1\u2642 , 1\u2640, same collection data; // Scenopinusjerei sp. nov. Pohjoism\u00e4ki & Haarto 2021 / (Diptera: Scenopinidae) / J. Pohjoism\u00e4ki det. // PARATYPE [yellow label] [MZH]; 1\u2642: Finland, EP: Isokyr\u00f6/ Orisberg 6983:[3]265 / 7.7.1999/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ vitripennis Meig./ det. A. Haarto 1999// PARATYPE/ Diptera: Scenopinidae/ Scenopinusjerei/ Pohjoism\u00e4ki & Haarto 2021 [red label] [AHC]; 1\u2642: Finland, ES [Etel\u00e4-Savo]: Rantasalmi/ Korhola 68720:[3]5802 / 27.6.2006/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ sp./ det. A. Haarto 2006// PARATYPE/ Diptera: Scenopinidae/ Scenopinusjerei/ Pohjoism\u00e4ki & Haarto 2021 [red label] [AHC]; 2\u2642: Finland, Kb: Ilomantsi/ Kelovaara 70008:36826 / 24.7.2021/ J. Pohjoism\u00e4ki leg. // PARATYPE/ Diptera: Scenopinidae/ Scenopinusjerei/ Pohjoism\u00e4ki & Haarto 2021 [yellow label] [JPC]; 1\u2640: Finland, ES: Rantasalmi/ Korhola 68720:5802 / 29.6.2006/ A. Haarto leg.// SCENOPINIDAE/ Scenopinus/ sp./ det. A. Haarto 2006// PARATYPE/ Diptera: Scenopinidae/ Scenopinusjerei/ Pohjoism\u00e4ki & Haarto 2021 [red label] [AHC]; 1\u2640: Finland, Kb: Liperi/ Viinij\u00e4rvi 6951:3615 / e larva 2013/ Ali Karhu leg.// linnunp\u00f6ntt\u00f6 [nest box]// SCENOPINIDAE/ Scenopinus/ sp./ det. A. Haarto 2014/ AHa14\u2013000891// PARATYPE/ Diptera: Scenopinidae/ Scenopinusjerei/ Pohjoism\u00e4ki & Haarto 2021 [red label] [AHC]; 1\u2640: Finland, Kb: Liperi/ K\u00e4s\u00e4m\u00e4 suo 6950:3619 / 26.-28.6.2013/ Ali Karhu leg.// SCENOPINIDAE/ Scenopinus/ sp./ det. A. Haarto 2020/ AHa20\u2013000473// PARATYPE/ Diptera: Scenopinidae/ Scenopinusjerei/ Pohjoism\u00e4ki & Haarto 2021 [red label] [MZT].Scenopinusjerei sp. nov. belongs to the S.fenestralis group and is easily recognisable from the other species in this group based on the contrasting colour differences between the femora and the yellow to orange tibiae. The coxae as well as the knob of the halteres are always uniformly black or dark brown, similar to the colour of the thorax.Male . ead Fig. . Frons brax Fig. . Very wemen Fig. . DorsallScenopinus spp. are poorly covered in the DNA barcode databases, such as Barcode of Life Database or GenBank. It is noteworthy that all S.fenestralis specimens in the databases from Europe to North America have almost identical COI sequences and represent the same barcode index number (BIN). The DNA barcode of Scenopinusjerei sp. nov. differs markedly from the other northern European species, its closest match being Scenopinusfenestralis from which it is separated by 12.48% sequence difference , especially Monopislaevigella (Denis & Schifferm\u00fcller), but also other Monopis spp., Niditineastriolella (Matsumura), and Tinea spp. Other insects observed from the same nest boxes included Ceratophyllus fleas, various beetles and flies . Apart for two male specimens found dead on a windowsill in an attic of an old house in Kelovaara on July 24 (see type specimens), most observations are from third week of June. According to the observations of Jere Kahanp\u00e4\u00e4 (pers. comm.), Scenopinusjerei sp. nov. hibernates as full-grown larvae and the adults emerge in a couple of weeks in room temperature rearing conditions. Based on the collection locations, it is likely that Scenopinusjerei sp. nov. is a boreal forest specialist.on the biology and distribution of Scenopinus spp., Scenopinusjerei sp. nov. is not very active flier, does not visit flowers and therefore is rarely collected by active netting or traps. Judging from the few Finnish observations, the species appears widespread in the southern and central parts of the country. We are certain that Scenopinusjerei sp. nov. can also be found in boreal forest biotopes in the other Nordic countries and Russia but has been until now overlooked.Like other This species is named after Mr. Jere Kahanp\u00e4\u00e4, Helsinki, who was to first to discover that the taxon is new to science and kindly agreed with the current arrangement for its formal description.Scenopinidae are outdated or difficult to obtain, we felt necessary to provide a key for the known European species of Scenopinidae. We must emphasise that we have been only able to examine the species with specimens listed in this paper, for which the identification key should work well. For the remainder, our approach was to go through the written species descriptions and pick features which we judged, by our collective species identification experience, to be useful for determination. To us this approach was better justified than reproducing the keys given in earlier literature, which are often difficult to follow or focus on limited number of poorly defined features. The diagnostic features for the key have been obtained from the descriptions in Because the existing literature on the European species of Scenopinusjerei sp. nov. was originally confused with Scenopinusvitripennis developing in cool and humid conditions are typically darker than the ones developed in warm and dry conditions . Legs entirely olive brown.Scenopinus pallipes Say, 1823 \u2013 White halteres, yellow legs.Scenopinus domesticus Meigen, 1824 \u2013 Legs yellow-red, head white from below.Scenopinus sulcicollis Meigen, 1824 \u2013 Legs yellow-red, head white from below, halteres white.Scenopinus scutellatus Macquart, 1843 \u2013 Halteres white, scutellum yellow.Scenopinus furcinervis Zetterstedt, 1844 \u2013 Legs fully yellow.Scenopinus fuscinervis Schiner, 1860 \u2013 The name is not mentioned in Schiner (1860).However, Kelsey attributes the synonymy to Schiner (1862), where S. fuscinervis Zetterstedt is given as a synonym of S. fenestralis. It is obvious that in this context, S. fuscinervis is a misspelling of S. furcinervis. In fact, the spelling is later corrected in Schiner (1864).Scenopinus graminicola Zetterstedt, 1859 \u2013 Halteres white.Scenopinus nigroscutellatus Frey, 1945 \u2013 From Azores, halteres white.Scenopinusjerei sp. nov. is not among the accepted species nor hidden among the synonyms of S.fenestralis. We hope that the species discovery reported here, together with the provisional indentification key we have provided, will encourage more research towards this exciting but poorly known family of flies.Based on this survey, we are confident that the species presented here as"} +{"text": "R. Soc. Open Sci.8, 201441. (Published online 21 April 2021) (doi:10.1098/rsos.201441)This correction refers to the caption for figure 2. An incorrect reference was given; the correct reference is [39]. This has now been corrected."} +{"text": "In this study, we critically revised and updated the checklist of native vascular plants of Mongolia. The checklist comprises 3,041 native vascular plant taxa from 653 genera and 111 families, including 7 lycophytes, 41 ferns, 21 gymnosperms, and 2,972 angiosperms. In the angiosperms, we identified the 14 families with the greatest species richness, ranging from 50 to 456 taxa. Species endemism is also noted here; 102 taxa are endemic to Mongolia, and 275 taxa are sub-endemic that co-occur in adjacent countries. Since 2014, a total of 14 taxa have been described new to science based on morphological evidences. Moreover, five genera and 74 taxa were newly added to the flora of Mongolia. Based on our critical revisions, names of three families, 21 genera, and 230 species have been changed in comparison to the previous checklist, \u201cConspectus of the vascular plants of Mongolia\u201d (2014). The flora of Mongolia is comprised of native species of different origins including boreal, steppe, desert, and mountainous elements of vegetation (Mongolia is located in the mid-latitude (between 41\u00b035'N\u201352\u00b009'N and 87\u00b044'E\u2013119\u00b056'E), between Russia and China, covering approximately 1.6 million kmgetation . The cougetation , namely,getation . Mongoligetation , which cgetation . Overallgetation . Howevergetation , which hCyperaceae (Apiaceae to Cornaceae (Huperziaceae to Ephedraceae (Asteraceae (Ceratophyllaceae to Zygophyllaceae (Amaranthaceae s.l. (incl. Chenopodiacceae) , Nymphaedelaceae .Brassicaceae family, the fifth-largest family in the country, was provided by Aquilegia L., Stipa L., and Primula L. were compiled by Geraniaceae in Mongolia. Orchidaceae, which included notes on their species richness and conservation status. The families Menyanthaceae and Nymphaeaceae were also revised by Additionally, several families and genera have been revised in recent years. For example, a new checklist of the Matthiola W.T.Aiton, Brassicaceae , for example, had two species that have been proven absent in the country due to the inaccurate location written on the herbarium specimens was found in northern Mongolia by Since sicaceae , Onocleacleaceae , Aldrovaseraceae , Hydrillritaceae , and Arcteraceae . Additiopecimens . On the In 2016, the orders and families of flowering plants were updated by the https://www.gbif.org/). We also compiled the phytogeographical regional distribution of all species, because species distribution is important information for species identification. The main herbaria for Mongolian flora , and all literature data for the species\u2019 regional distribution, have been checked and studied. The regional distribution of the taxa mostly follows The systematic order and taxonomic circumscription of the families is based on the following classifications: Ferns and Fern Allies by The current checklist comprises 3,042 native vascular plant taxa , belonging to 653 genera and 111 families (Table Asteraceae (85 genera and 456 taxa), Fabaceae (24 and 328), Poaceae (58 and 229), Rosaceae (28 and 168), Ranunculaceae (20 and 156), Brassicaceae (51 and 138), Cyperaceae (10 and 130), Lamiaceae (22 and 103), Amaranthaceae (30 and 94), Caryophyllaceae (20 and 97), Boraginaceae (24 and 78), Apiaceae (36 and 66), Polygonaceae (11 and 63), and Orobanchaceae (9 and 57) , Artemisia L. (103), Carex L. (99) Oxytropis DC. (97), Potentilla L. (75), Saussurea DC. (55), Taraxacum F.H.Wigg. (53), Allium L. (50), Salix L. (42), Ranunculus L. (41), Pedicularis L. (36), Poa L. (28), Viola L. (27), and Silene L. (24) which is shown in Fig. There are 14 families with a high species richness (\u2265 9 genera and \u2265 57 taxa): Fabaceae (74 taxa) show the highest number of sub-endemic taxa along with Asteraceae (60 taxa), Brassicaceae (23 taxa), Poaceae (18 taxa), and Amaranthaceae (9 taxa). The highest number of sub-endemic taxa were found in the Mongolian Altai (114 taxa) followed by Khangai (87 taxa), Gobi-Altai (76 taxa), Khovd (68 taxa), Khuvsgul (63 taxa), and the Depression of Great Lakes (60 taxa). The remaining ten regions have between 24 and 58 sub-endemic taxa were identified: the Mongolian Altai , Khangai , Khentei 1,236 taxa), Mongolian Dauria , Khuvsgul , and Khovd . The remaining ten regions have between 262 and 952 taxa and Portulacaceae (Portulacaoleracea L.). Several taxa were found to be archeophytes, which were introduced in \u201cancient\u201d times and became naturalized as part of the native flora and Portulacaceae (Portulacaoleracea L.) and 57 other taxa . Moreover, approximately 19,300 images of 1,780 taxa have been observed as part of citizen science contributions to the \u201cFlora of Mongolia\u201d project on the iNaturalist platform , which was established on January 2019.Both of the online databases are allowing researchers to collaborate and revise Mongolian taxa more readily and will continue to improve the documentation of Mongolia\u2019s flora. To date, a total of 2,249 taxa (ca. 73% of the flora) have been deposited in the database of In this study, we checked more than 70 works published since 2013 that have revised the flora of Mongolia, and provided respective references for each species in our checklist. We reviewed the species status of all vascular flora of Mongolia and made critical changes by adding, synonymizing, and excluding taxa; this work resulted in 265 fewer taxa compared to SE].The families in the checklist are alphabetically ordered and, within them, the genera, species, and subspecies are alphabetically listed. The currently accepted names are highlighted in bold italics. The most common synonyms (previously used in I LycophytesLycopodiaceae P.Beauv. (3 genera and 5 species)1. Diphasiastrumalpinum (L.) Holub Diphasiastrumcomplanatum (L.) Holub [1]Huperziaselago (L.) Bernh. Lycopodiumannotinum L. Lycopodiumclavatum L. [2]Selaginellaceae2. Willk. (1 genus and 2 species)Selaginellaborealis (Kaulf.) Spring Selaginellasanguinolenta (L.) Spring II Ferns and fern alliesAspleniaceae3. Newman (1 genus and 5 species)Aspleniumaltajense (Kom.) Grubov Aspleniumruprechtii Sa.Kurata Aspleniumruta-muraria L. Aspleniumseptentrionale (L.) Hoffm. Aspleniumyunnanense Franch. Athyriaceae4. Alston (2 genera and 4 species)Athyriumfilix-femina (L.) Roth [1\u20135]Athyriummonomachi Kom. Athyriumsinense Rupr. Diplaziumsibiricum (Turcz.) Sa.Kurata [1\u20135]Cystopteridaceae5. Shmakov (2 genera and 4 species)Cystopterisfragilis (L.) Bernh. Cystopterissudetica A.Braun & Milde [2]Gymnocarpiumdryopteris Newman Gymnocarpiumjessoense (Koidz.) Koidz. Dennstaedtiaceae6. Losty (1 genus and 1 species)Pteridiumaquilinum (L.) Kuhn Dryopteridaceae7. Herter (1 genus and 3 species)Dryopterisdilatata (Hoffm.) A.Gray Dryopterisexpansa (C.Presl) Fraser-Jenk. & Jermy Dryopterisfragrans (L.) Schott Equisetaceae8. Michx. (1 genus and 9 species)Equisetumarvense L. Equisetumfluviatile L. Equisetumhyemale L. Equisetumpalustre L. Equisetumpratense Ehrh. Equisetumramosissimum Desf. [14]Equisetumscirpoides Michx. Equisetumsylvaticum L. Equisetumvariegatum Schleich. Onocleaceae9. Pic.Serm. (2 genera and 2 species)Matteucciastruthiopteris (L.) Tod. Onocleasensibilis L. [2]Ophioglossaceae10. Martinov (1 genus and 2 species)Botrychiumlanceolatum (Gmel.) \u00c5ngstr. [3]Botrychiumlunaria (L.) Sw. Polypodiaceae11. J.Presl. & C.Presl (2 genera and 2 species)Lepisorusclathratus Ching [13]Polypodiumvirginianum L. Pteridaceae12. E.D.M.Kirchn. (2 genera and 2 species)Cheilanthesargentea (S.G.Gmel.) Kunze Cryptogrammastelleri (S.G.Gmel.) Prantl [1]Thelypteridaceae13. Ching (1 genus and 1 species)Phegopterisconnectilis (Michx.) Watt Woodsiaceae14. Herter (1 genus and 6 species)Woodsiacalcarea (Fomin) Shmakov Woodsiaglabella R.Br. Woodsiaheterophylla (Turcz.) Shmakov [1]Woodsiailvensis (L.) R.Br. [= Woodsiaacuminata (Fomin) Sipliv.] Woodsiapseudopolystichoides (Fomin) Kiselev & Shmakov [5]Woodsiasubcordata Turcz. III GymnospermsCupressaceae15. Gray (1 genus and 4 taxa)Juniperuscommunis L. Juniperuspseudosabina Fisch. & C.A.Mey. Juniperussabinavar.davurica Farjon [= Juniperusdavurica Pall.] [2]JuniperussabinaL.var.sabina Ephedraceae16. Dumort. (1 genus and 9 species)Ephedradahurica Turcz. [= Ephedrasinicasubsp.dahurica (Turcz.) Galanin] Ephedraequisetina Bunge Ephedrafedtschenkoi Paulsen Ephedraglauca Regel Ephedraintermedia Schrenk & C.A.Mey. Ephedralomatolepis Schrenk Ephedramonosperma J.G.Gmel. Ephedraprzewalskii Stapf Ephedrasinica Stapf Pinaceae17. Spreng. (4 genera and 8 species)Abiessibirica Ledeb. Larixczekanowskii Szafer [4]Larixgmelinii (Rupr.) Kuzen. [\u2261 Abiesgmelinii Rupr.] Larixsibirica Ledeb. Piceaobovata Ledeb. Pinuspumila Regel [2]Pinussibirica Du Tour Pinussylvestris L. IV AngiospermsAcoraceae18. Martinov (1 genus and 1 species)Acoruscalamus L. Adoxaceae19. E.Mey. [including Viburnaceae Raf.] (3 genera and 6 species)Adoxamoschatellina L. Sambucussibirica Nakai Sambucuswilliamsii Hance [= Sambucusmanshurica Kitag.] Viburnumburejaeticum Regel & Herder [5]Viburnummongolicum Rehder [= Loniceramongolica Pall.] Viburnumsargentii Koehne [5]Alismataceae20. Vent. (2 genera and 4 species)Alismagramineum Lej. Alismaplantago-aquatica L. Sagittarianatans Pall. Sagittariatrifolia L. Amaranthaceae21. Juss. [including Chenopodiaceae Vent.] (34 genera and 94 taxa)Agriophyllumpungens (Vahl) Link [= Agriophyllumsquarrosum Moq.] [6\u201316]Anabasisaphylla L. Anabasisbrevifolia C.A.Mey. Anabasiselatior (C.A.Mey.) Schischk. [14]Anabasiseriopoda Paulsen [14]Anabasispelliotii Danguy [14]Anabasissalsa Paulsen [14]Anabasistruncata Bunge Atriplexaltaica Sukhor. [7]Atriplexcana C.A.Mey. [14]Atriplexfera (L.) Bunge Atriplexlaevis C.A.Mey. Atriplexsibirica L. Atriplextatarica L. Axyrisamaranthoides L. Axyrishybrida L. Axyrisprostrata L. Bassiahyssopifolia Kuntze Bassiaprostrata (L.) Beck [\u2261 Kochiaprostrata (L.) Schrad.] [1\u201315]Bassiascoparia (L.) A.J.Scott [\u2261 Kochiascoparia (L.) Schrad.] Blitumvirgatum L. [\u2261 Chenopodiumfoliosum Asch.] Camphorosmamonspeliacasubsp.lessingii (Litv.) Aellen Caroxylongemmascens Tzvelev [\u2261 Salsolagemmascens Pall.] [10]Caroxylonpasserinum (Bunge) Akhani & Roalson [\u2261 Salsolapasserina Bunge] Ceratocarpusarenarius L. Chenopodiastrumhybridum (L.) S.Fuentes, Uotila & Borsch [\u2261 Chenopodiumhybridum L.] Chenopodiumacuminatum Willd. Chenopodiumalbum L. [1\u201316]Chenopodiumficifolium Sm. Chenopodiumfrutescens C.A.Mey. Chenopodiumiljinii Golosk. Chenopodiumkaroi Aellen [1\u201315]Chenopodiumnovopokrovskyanum (Aellen) Uotila [\u2261 Chenopodiumalbumsubsp.novopokrovskyanum (Aellen) Uotila] [7]Chenopodiumstrictum Roth Chenopodiumvulvaria L. Climacopteraaffinis (C.A.Mey.) Botsch. [\u2261 Pyankoviaaffinis (C.A.Mey.) Mosyakin & Roalson] [14]Climacopterasubcrassa (Popov) Botsch. [14]Corispermumchinganicum Iljin [1\u201312]Corispermumdeclinatum Steph. ex Iljin Corispermumelongatum Bunge [\u2261 Corispermumstauntoniisubsp.elongatum (Bunge) Vorosch.] Corispermummongolicum Iljin SECorispermumpatelliforme Iljin SECorispermumtylocarpum Hance [= Corispermumgmelinii Bunge] [12]Dysphaniabotrys (L.) Mosyakin & Clemants Gruboviadasyphylla (Fisch. & C.A.Mey.) Freitag & G.Kadereit [3\u201316]Gruboviakrylovii (Litv.) Freitag & G.Kadereit [\u2261 Kochiakrylovii Litv.] Gruboviamelanoptera (Bunge) Freitag & G.Kadereit [\u2261 Kochiamelanoptera Bunge] Halocnemumstrobilaceum M.Bieb. [14]Halogetonglomeratus (M.Bieb.) C.A.Mey. Halostachyscaspica C.A.Mey. Haloxylonammodendron (C.A.Mey.) Bunge Iljiniaregelii (Bunge) Korovin Kalidiumcaspicum (L.) Ung.-Sternb. Kalidiumcuspidatum (Ung.-Sternb.) Grubov Kalidiumfoliatum Moq. SEKalidiumgracile Fenzl Krascheninnikoviaceratoides (L.) Gueldenst. [= Krascheninnikoviaewersmanniana (Stschegl.) Grubov] Micropeplisarachnoidea (Moq.) Bunge [\u2261 Halogetonarachnoides Moq.] SENanophytongrubovii U.P.Pratov [10]SENanophytonmongolicum U.P.Pratov Oreosalsolaabrotanoides (Bunge) Akhani [\u2261 Salsolaabrotanoides Bunge] [6\u201313]Oxybasischenopodioides (L.) S.Fuentes, Uotila & Borsch [\u2261 Chenopodiumchenopodioides (L.) Aellen] Oxybasisglauca (L.) S.Fuentes, Uotila & Borsch [\u2261 Chenopodiumglaucum L.] [2\u201316]Oxybasisgubanovii (Sukhor.) Sukhor. & Uotila [\u2261 Chenopodiumgubanovii Sukhor.] Oxybasisrubra (L.) S.Fuentes, Uotila & Borsch [\u2261 Chenopodiumrubrum L.] Oxybasisurbica (L.) S.Fuentes, Uotila & Borsch [\u2261 Chenopodiumurbicum L.] Petrosimonialitvinowii Korsh. [10]Petrosimoniasibirica Bunge [14]SESalicorniaaltaica Lomon. [\u2261 Salicorniaperennanssubsp.altaica (Lomon.) G.Kadereit & Piirainen] [7]Salsolacollina Pall. [\u2261 Kalicollinum Akhani & Roalson] [2\u201315]SESalsolaikonnikovii Iljin [\u2261 Kaliikonnikovii (Iljin) Akhani & Roalson] Salsolajacquemontii Moq [\u2261 Kalijacquemontii (Moq.) Akhani & Roalson] Salsolalaricifolia Litv. Salsolamonoptera Bunge [\u2261 Kalimonopterum (Bunge) Lomon.] Salsolapaulsenii Litv. [\u2261 Kalipaulsenii (Litv.) Akhani & Roalson] Salsolarosacea L. Salsolatragus L. Sodafoliosa (L.) Akhani [\u2261 Salsolafoliosa L. \u2261 Neocaspiafoliosa (L.) Tzvelev] [14]Suaedaacuminata (C.A.Mey). Moq. Suaedacorniculata(C.A.Mey.)Bungesubsp.corniculata Suaedacorniculatasubsp.mongolica Lomon. & Freitag Suaedaglauca (C.A.Mey.) Bunge Suaedaheterophylla (Kar. & Kir.) Bunge [10\u201315]Suaedakossinskyi Iljin [\u2261 Bienertiakossinskyi (Iljin) Tzvelev] Suaedalinifolia Pall. Suaedaprostrata Pall. [= Suaedamaritima auct. non L.] Suaedaprzewalskii Bunge [\u2261 Bienertiaprzewalskii (Bunge) G.L.Chu] [10\u201313]Suaedasalsa (L.) Pall. Suaedasibirica Lomon. & Freitag SESuaedatschujensis Lomon. & Freitag SESuaedatuvinica Lomon. & Freitag Sympegmaregelii Bunge Teloxysaristata (L.) Moq. [\u2261 Chenopodiumaristatum L. \u2261 Dysphaniaaristata (L.) Mosyakin & Clemants] Xylosalsolaarbuscula Tzvelev [\u2261 Salsolaarbuscula Pall.] Amaryllidaceae22. J.St.-Hil. (1 genus and 50 taxa)Alliumspirale is absent in Mongolia and A.subangulatum was found in southern Gobi by Note: According to Alliumaltaicum Pall. Alliumamphibolum Ledeb. Alliumanisopodium Ledeb. [2\u201313]SEAlliumaustrosibiricum N.Friesen Alliumbaicalense Willd. [= Alliumsenescenssubsp.glaucum (Schrader) Dost\u00e1l] Alliumbidentatum Fisch. Alliumburjaticum N.Friesen Alliumcarolinianum Redout\u00e9 [14]Alliumchamarense M.M.Ivanova Alliumclathratum Ledeb. Alliumcondensatum Turcz. Alliumeduardi Stearn Alliumflavidum Ledeb. Alliumgalanthum Kar. & Kir. Alliumhymenorrhizum Ledeb. Alliumkarelinii Poljakov Alliumledebourianum Schult. & Schult.f. [7]Alliumleucocephalum Turcz. Alliummacrostemon Bunge Alliummalyschevii N.Friesen Alliummaximowiczii Regel Alliummicrodictyon Prokh. Alliummonadelphum Turcz. Alliummongolicum Regel Alliumneriniflorum G.Don Alliumobliquum L. [7]Alliumoliganthum Kar. & Kir. Alliumpallasii Murray [14]AlliumplatyspathumSchrenksubsp.platyspathum Alliumplatyspathumsubsp.amblyophyllum (Kar. & Kir.) N.Friesen Alliumpolyrhizum Turcz. Alliumprostratum Trev. [1\u201313]SEAlliumpumilum Vved. Alliumramosum L. [1\u201313]Alliumrubens Schrad. Alliumschischkinii Sobolevsk. Alliumschoenoprasum L. Alliumschrenkii Regel [= Alliumbogdoicola Regel] Alliumsenescens L. Alliumsplendens Willd. Alliumspurium G.Don Alliumstellerianum Willd. Alliumstrictum Schrad. SEAlliumsubangulatum Regel [16]Alliumsubtilissimum Ledeb. Alliumtenuissimum L. Alliumtuvinicum (N.Friesen) N.Friesen [\u2261 Alliumstellerianumsubsp.tuvinicum N.Friesen] SEAlliumtytthocephalum Schult.f. SEAlliumubsicola Regel Alliumvodopjanovae N.Friesen Apiaceae23. Lindl. (36 genera and 66 taxa)Aegopodiumalpestre Ledeb. Angelicaczernaevia (Fisch. & C.A.Mey.) Kitag. Angelicadahurica (Hoffm.) Benth. & Hook.f. [\u2261 dahurica Hoffm.] Angelicasaxatilis Turcz. [\u2261 Physolophiumsaxatile (Turcz.) Turcz.] [2]Angelicasylvestris L. Anthriscussylvestris (L.) Hoffm. [1\u201310]Archangelicadecurrens Ledeb. [\u2261 Angelicaarchangelicasubsp.decurrens (Ledeb.) Kuvaev] Aulacospermumanomalum Ledeb. Bupleurumaureum Fisch. [7]Bupleurumbicaule Helm [= Bupleurumpusillum Krylov] Bupleurumdensiflorum Rupr. [= Bupleurummongolicum V.M.Vinogr.] Bupleurumkrylovianum Schischk. Bupleurummultinerve DC. [= Bupleurumlongeinvolucratum Krylov] Bupleurumscorzonerifolium Willd. Bupleurumsibiricum Vest Carumburiaticum Turcz. Carumcarvi L. Cenolophiumdenudatum (Hornem.) Tutin Cicutavirosa L. [1\u201315]Cnidiumdauricum (Jacq.) Turcz. [\u2261 Laserpitiumdauricum Jacq.] [2\u201310]Cnidiummonnieri Cusson SEConioselinumlongifolium Turcz. Conioselinumtataricum Hoffm. [= Conioselinumvaginatum (Spreng.) Thell.] Elwendiasetacea (Schrenk) Pimenov & Kljuykov SEFerulabungeana Kitag. Ferulacaspica M.Bieb. Feruladissecta Ledeb. Feruladshaudshamyr Korovin [= Feruladubjanskyi Korovin] Ferulaferulioides (Steud.) Korovin [7]Ferulapotaninii Korovin [14]Ferulasoongarica Pall. [= Ferulamongolica (V.M.Vinogr. & Kamelin) V.M.Vinogr. & Kamelin] SEFerulopsishystrix (Bunge) Pimenov [\u2261 Peucedanumhystrix Bunge] SEHaloselinumfalcaria (Turcz.) Pimenov [\u2261 Peucedanumfalcaria Turcz.] Hanseniamongholica Turcz. [\u2261 Ligusticummongholicum (Turcz.) Krylov] Heracleumdissectum Ledeb. Heracleumsibiricum L. Kadeniasalina (Turcz.) Lavrova & V.N.Tikhom. [\u2261 Cnidiumsalinum Liou] Kitagawiabaicalensis (Redow.) Pimenov [\u2261 Peucedanumbaicalense (Redow.) Koch] Kitagawiaterebinthacea (Fisch.) Pimenov [\u2261 Peucedanumterebinthaceum (Fisch.) Ledeb.] SELithosciadiumkamelinii (V.M.Vinogr.) Pimenov [\u2261 Cnidiumkamelinii V.M.Vinogr.] [7]SELithosciadiummulticaule Turcz. Neogayasimplex Meisn. [= Pachypleurumalpinum Ledeb.] [10]Oenantheaquatica (L.) Poir. [= Peucedanumsalinum Pall.] Ostericumtenuifolium Y.C.Chu [= Pachypleurumalpinum Ledeb.] Paraligusticumdiscolor (Ledeb.) V.N.Tikhom. [7]SEPeucedanumpuberulum Turcz. Peucedanumvaginatum Ledeb. Phlojodicarpussibiricus Koso-Pol. Phlojodicarpusvillosus Turcz. Pimpinellathellungiana H.Wolff Pleurospermumuralense Hoffm. Prangosledebourii Herrnst. & Heyn Sajanellamonstrosa (Willd.) Soj\u00e1k Saposhnikoviadivaricata (Turcz.) Schischk. Schulziacrinita Spreng. Seseliabolinii (Korovin) Schischk. [\u2261 Libanotisabolinii (Korovin) Korovin] Seselibuchtormense W.D.J.Koch [\u2261 Libanotisbuchtormensis (Fisch.) DC.] Seselicondensatum Rchb.f. [\u2261 Libanotiscondensata (L.) Fisch.] Seselieriocarpum B.Fedtsch. [\u2261 Libanotiseriocarpa Schrenk] Seseliglabratum Willd. [= Libanotistenuifolia DC.] [7]SESeseligrubovii V.M.Vinogr. & Sanchir [\u2261 Libanotisgrubovii (V.M.Vinogr. & Sanchir) M.L.Sheh & M.F.Watson] Seselimucronatum (Schrenk) Pimenov & Sdobnina [14]Seseliseseloides (Fisch. & C.A.Mey.) M.Hiroe [\u2261 Libanotisseseloides (Fisch. & C.A.Mey.) Turcz.] Siumsuave Walter Sphallerocarpusgracilis Koso-Pol. Stenocoeliumathamantoides Ledeb. [\u2261 Seseliathamantoides (M.Bieb.) Beck] Apocynaceae24. Juss. (3 genera and 10 taxa)Apocynumpictum Schrenk [= Apocynumhendersonii Hook.f.] Apocynumvenetum L. [\u2261 Poacynumvenetum (L.) Mavrodiev] [14]Cynanchumacutumsubsp.sibiricum (Willd.) Rech.f. [10\u201316]Cynanchumbungei Decne. [9]Cynanchumchinense R.Br. SECynanchumgobicum Grubov [= Vincetoxicumlanceolatum (Grubov) Grubov] [12\u201316]Cynanchummongolicum Hemsl. [16]Cynanchumpurpureum K.Schum. Vincetoxicummukdenense Kitag. [= Cynanchumpaniculatum (Bunge) Kitag.] Vincetoxicumsibiricum (L.) Decne. [= Cynanchumthesioides K.Schum.] [2\u201316]Araceae25. Juss. (2 genera and 4 species)Lemnaminor L. Lemnatrisulca L. Lemnaturionifera Landolt Spirodelapolyrhiza (L.) Schleid. Asparagaceae26. Juss. (5 genera and 19 species)Anemarrhenaasphodeloides Bunge Asparagusbrachyphyllus Turcz. [9]SEAsparagusburjaticus Peschkova [4]Asparagusdauricus Fisch. SEAsparagusgobicus Ivanova [7\u201316]Asparagusneglectus Kar. & Kir. [14]Asparagusoligoclonos Maxim. [5]Asparaguspallasii Miscz. Asparagusschoberioides Kunth [5]Asparagustamariscinus Ivanova Asparagustrichophyllus Bunge Convallariakeiskei Miq. Maianthemumbifolium (L.) F.W.Schmidt Maianthemumdilatatum (Alph.Wood) A.Nelson & J.F.Macbr. Maianthemum\u00d7intermedium Vorosch. [5]Maianthemumtrifolium (L.) Sloboda [2]Polygonatumhumile Fisch. Polygonatumodoratum (Mill.) Druce Polygonatumsibiricum Redout\u00e9 Asphodelaceae27. Juss. [including Xanthorrhoeaceae Dumort.] (1 genus and 2 taxa)HemerocallislilioasphodelusL.var.lillioasphodelus Hemerocallislilioasphodelusvar.minor (Mill.) M.N.Tamura [\u2261 Hemerocallisminor Mill.] Asteraceae28. Bercht. & J.Presl (85 genera and 456 taxa)Asteraceae have changed after extnesive molecular investigations. For example, species of Scorzonera L. were split into several genera, and three of them are present in Mongolia: Lipschitzia Zaika, Sukhor. & N.Kilian, Takhtajaniantha Nazarova, and Scorzonera L. s.str. by Scorzoneracurvata, S.grubovii, and S.sinensis is not resolved yet.Note: Some classifications of some genera of Achilleaacuminata Sch.Bip. Achilleaalpina L. Achilleaasiatica Serg. Achilleaimpatiens L. Achillealedebourii Heimerl Achilleamillefolium L. Achilleaptarmicoides Maxim. Achilleasergievskiana Shaulo & Shmakov [7]SEAjaniaachilleoides Poljakov Ajaniafruticulosa (Ledeb.) Poljakov Ajaniagrubovii Muldashev [\u2261 Chrysanthemumgrubovii (Muldashev) H.Ohashi & Yonek.] E Ajaniatrifida (Turcz.) Muldashev [\u2261 Hippolytiatrifida (Turcz.) Poljakov] Allardiatridactylites Sch.Bip. [\u2261 Waldheimiatridactylites Kar. & Kir.] Ancathiaigniaria DC. Antennariadioica (L.) Gaertn. Arctiumtomentosum Mill. [4]Arctogerongramineum (L.) DC. Arnicaangustifoliasubsp.iljinii (Maguire) I.K.Ferguson [7]Artemisiaadamsii Besser SEArtemisiaaksaiensis Y.R.Ling Artemisiaamoena Poljakov Artemisiaanethifolia Weber Artemisiaanethoides Mattf. [8\u201316]Artemisiaannua L. Artemisiaargyi H.L\u00e9v. & Vaniot Artemisiaargyrophylla Ledeb. Artemisiaassurgens Filatova [\u2261 Seriphidiumassurgens (Filatova) K.Bremer & Humphries] E Artemisiaaurata Kom. Artemisiabargusinensis Spreng. SEArtemisiablepharolepis Bunge Artemisiaborealis Pall. Artemisiaborotalensis Poljakov [\u2261 Seriphidiumborotalense (Poljakov) Ling & Y.R.Ling] SEArtemisiabrachyloba Franch. Artemisiabrachyphylla Kitam. [5]Artemisiacaespitosa Ledeb. Artemisiacapillaris Thunb. Artemisiacompacta Fisch. Artemisiadahurica (Turcz.) Poljakov [4]Artemisiadavazamczii Darijma & Kamelin E Artemisiademissa Krasch. Artemisiadepauperata Krasch. ArtemisiadesertorumSpreng.subsp.desertorum Artemisiadesertorumsubsp.pseudojaponica Darijma & Kamelin [5]E SEArtemisiadisjuncta Krasch. SEArtemisiadolosa Krasch. SEArtemisiadracunculusvar.changaica (Krasch.) Y.R.Ling [\u2261 Artemisiachangaica Krasch.] ArtemisiadracunculusL.var.dracunculus [1\u201315]Artemisiaeriopoda Bunge [16]Artemisiafeddeisubsp.arschantinica (Darijma) Gubanov & Kamelin [\u2261 Artemisiaarschantinica Darijma] [16]ArtemisiafeddeiH.L\u00e9v. & Vaniotsubsp.feddei Artemisiafreyniana (Pamp.) Krasch. Artemisiafrigida Willd. [1\u201316]SEArtemisiagiraldii Pamp. [4]Artemisiaglauca Pall. SEArtemisiaglobosa Krasch. SEArtemisiaglobosoides Ling & Y.R.Ling ArtemisiagmeliniiWeb.var.gmelinii [2\u201313]Artemisiagmeliniivar.messerschmidiana (Besser) Poljakov Artemisiagracilescens Krasch. & Iljin Artemisiahalodendron Turcz. Artemisiaheptapotamica Poljakov [\u2261 Seriphidiumheptapotamicum (Poljakov) Ling & Y.R.Ling] Artemisiaimplicata T.G.Leonova [16]Artemisiaintegrifolia L. Artemisiaklementzae Krasch. [= Artemisiaxylorhiza Krasch.] Artemisialaciniata Willd. SEArtemisialagocephalaFisch.var.lithophila (Turcz.) Y.R.Ling [1]Artemisialatifolia Ledeb. Artemisiamacilenta (Maxim.) Krasch. Artemisiamacrantha Ledeb. Artemisiamacrocephala Jacquem. [1\u201316]Artemisiamanshurica (Kom.) Kom. Artemisiamarschalliana Spreng. [7]Artemisiamaximovicziana Krasch. Artemisiamedioxima Krasch. Artemisiamongolica (Fisch.) Nakai [\u2261 Artemisiavulgarisvar.mongolica Fisch.] [1\u201315]Artemisiamongolorumsubsp.gobicum Krasch. [\u2261 Artemisiagobica (Krasch.) Grubov] ArtemisiamongolorumKrasch.subsp.mongolorum [\u2261 Seriphidiummongolorum (Krasch.) Ling & Y.R.Ling] Artemisianitrosa Weber Artemisiaobtusilobasubsp.altaiensis (Krasch.) Krasnob. [\u2261 Artemisiaaltaiensis Krasch.] ArtemisiaobtusilobaLedeb.subsp.obtusiloba Artemisiaobtusilobavar.glabra Ledeb. [= Artemisiaglabella Kar. & Kir.] SEArtemisiaordosica Krasch. SEArtemisiaoxycephala Kitag. Artemisiapalustris L. [1\u201313]Artemisiapamirica C.Winkl. Artemisiaphaeolepis Krasch. Artemisiapubescens Ledeb. [= Artemisiacommutata Besser] Artemisiapycnorrhiza Ledeb. Artemisiarubripes Nakai Artemisiarupestris L. Artemisiarutifoliavar.altaica (Krylov) Krasch. [7]Artemisiasacrorumvar.messerschmidtiana (Besser) Y.R.Ling Artemisiasaissanica (Krasch.) Filatova Artemisiasantolinifolia Turcz. [= Artemisiasantolinifoliasubsp.stepposa Darijma] Artemisiaschischkinii Krasch. Artemisiaschrenkiana Ledeb. Artemisiascoparia Waldst. & Kit. [2\u201312]Artemisiaselengensis Turcz. Artemisiasericea Weber Artemisiasieversiana Ehrh. [1\u201316]SEArtemisiasphaerocephala Krasch. Artemisiastolonifera (Maxim.) Kom. SEArtemisiasubchrysolepis Filatova [\u2261 Seriphidiumsubchrysolepis (Filatova) K.Bremer & Humphries] Artemisiasubdigitata Mattf. [\u2261 Artemisiadubiavar.subdigitata (Mattf.) Y.R.Ling] Artemisiasublessingiana Krasch. [= Seriphidiumgorjaevii (Poljak.) Y.R.Ling] [14]Artemisiasubulata Nakai Artemisiasucculenta Ledeb. [7]Artemisiasylvatica Maxim. Artemisiatanacetifolia L. Artemisiaterrae-albae Krasch. Artemisiatomentella Trautv. Artemisiatournefortiana Rchb. SEArtemisiatransbaicalensis T.G.Leonova Artemisiaumbrosa (Besser) Turcz. Artemisiavestita Wall. [13]Artemisiaviridis Willd. Artemisiavulgarissubsp.vulgaris L. Artemisiavulgarissubsp.inundata Darijma [= Artemisiasuperba Pamp.] E Artemisiawudanica Liou & W.Wang SEArtemisiaxanthochloa Krasch. [3\u201316]Artemisiaxerophytica Krasch. Askelliaflexuosa (Ledeb.) W.A.Weber Askelliapygmaea (Ledeb.) Sennikov Asteralpinus L. Asterhispidus Thunb. Asterlingii G.J.Zhang & T.G.Gao [= Rhinactinidialimoniifolia Novopokr.] [7]Astermaackii Regel [5]Astersanczirii Kamelin & Gubanov [5]E Astertataricus L.f. SEAsterothamnusalyssoides (Turcz.) Novopokr. [= Asteralyssoides Turcz.] SEAsterothamnuscentraliasiaticusvar.potaninii (Novopokr.) Y.Ling & Y.L.Chen [\u2261 Asterothamnuspotaninii Novopokr.] SEAsterothamnusheteropappoides Novopokr. Asterothamnusmolliusculus Novopokr. Asterothamnuspoliifolius Novopokr. Bidenscernua L. Bidensparviflora Willd. Bidensradiata Thuill. Bidenstripartita L. SEBrachanthemumgobicum Krasch. SEBrachanthemummongolicum Krasch. Brachanthemummongolorum Grubov [9]E Cancriniadiscoidea (Ledeb.) Poljakov SECancriniakrasnoborovii Khanm. [10]Carduuscrispus L. Carduusnutans L. Centaureaadpressa Ledeb. [6]Centaureaglastifoliasubsp.intermedia (Boiss.) L.Martins [= Centaureachartolepis Greuter] Centaureapulchella Ledeb. [= Hyaleapulchella (Ledeb.) K.Koch] Chondrillalejosperma Kar. & Kir. Chrysanthemumchalchingolicum Grubov E Chrysanthemummongolicum Ling [\u2261 Chrysanthemumzawadzkiivar.mongolicum (Ling) Gubanov] Chrysanthemumnaktongense Nakai [9]SEChrysanthemumsinuatum Ledeb. [Tanacetumsinuatum Sch.Bip.] Chrysanthemumtrilobatum (Poljakov) H.Ohashi & Yonek. [\u2261 Ajaniatrilobata Poljakov] Chrysanthemumzawadzkii Herbich Cicerbitaazurea (Ledeb.) Beaverd Cirsiumarvense (L.) Scop. [\u2261 Serratulaarvensis L.] Cirsiumesculentum C.A.Mey. Cirsiumglabrifolium O.Fedtsch. & B.Fedtsch. [7]Cirsiumhelenioides (L.) Hill [= Carduushelenioides L.] [2]Cirsiumpendulum Fisch. Cirsiumserratuloides Hill Cirsiumsetosum (Willd.) M.Bieb. [\u2261 Serratulasetosa Willd.] Cirsiumsieversii (Fisch. & C.A.Mey.) Petr. [= Cirsiumpolyacanthum Kar. & Kir.] [7]Cirsiumvlassovianum Fisch. Cousiniaaffinis Schrenk [14]Crepidiastrumakagii (Kitag.) J.W.Zhang & N.Kilian [= Youngiatenuicaulis (Babc. & Stebbins) Czerep.] Crepidiastrumsonchifolium (Bunge) Pak & Kawano [5]Crepidiastrumtenuifolium (Willd.) Sennikov [\u2261 Crepistenuifolia Willd. \u2261 Youngiatenuifolia (Willd.) Babc. & Stebbins] Crepisbungei Ledeb. Crepischrysantha Froel. Crepiscrocea(Lam.)Babc.var.crocea SECrepiscroceavar.czuensis (Serg.) Tzvelev [\u2261 Crepisczuensis Serg.] Crepislomonosovae Tzvelev E Crepislyrata (L.) Froel. Crepismulticaulis Ledeb. Crepispolytricha Turcz. Crepispraemorsa (L.) Tausch [\u2261 Hieraciumpraemorsum L.] Crepissibirica L. Crepistectorum L. Doronicumaltaicum Pall. [1]Doronicumoblongifolium DC. Doronicumturkestanicum Cavill. Echinopsdavuricus Fisch. [= Echinopslatifolius Tausch] Echinopsgmelinii Turcz. Echinopshumilis M.Bieb. Echinopsintegrifolius Kar. & Kir. Echinopsnanus Bunge Echinopsritro L. Erigeronacris L. Erigeronaltaicus Popov SEErigeronbaicalensis Botsch. [1]Erigeroneriocalyx (Ledeb.) Vierh. Erigeronkrylovii Serg. Erigeronlonchophyllus Hook. Erigeronoreades Fisch. & C.A.Mey. Erigeronpetiolaris Vierh. Erigeronpolitus Fr. Erigeronpseudoeriocephalus Popov [3]Filagoarvensis L. Filifoliumsibiricum (L.) Kitam. [= Tanacetumsibiricum L.] Galatellaaltaica Tzvelev Galatellaangustissima (Tausch) Novopokr. [1]Galatelladahurica DC. [= Galatellamacrosciadia Gand. = Galatellasongorica Novopokr.] Galatellahauptii Lindl. [7]Gnaphaliumuliginosum L. [= Gnaphaliumbaicalense Kirp. & Kuprian.] Helichrysumarenarium Moench [7]Heteropappusaltaicus Novopokrov. [\u2261 Asteraltaicus Willd.] Heteropappusbiennis (Ledeb.) Tamamsch. SEHeteropappusmedius (Krylov) Tamamsch. SEHieraciumczadanense Tupitz. Hieraciumkorshinskyi Zahn Hieraciumnarymense Schischk. & Serg. Hieraciumrobustum Fr. Hieraciumsershukense \u00dcksip [7]Hieraciumsubramosum Lonnr. Hieraciumumbellatum L. Hieraciumvirosum Pall. Hololeionmaximowiczii Kitam. [9]Hypochaerismaculata L. [\u2261 Trommsdorffiamaculata (L.) Bernh.] Inulajaponica Thunb. [2]Inulalinariifolia Turcz. Inulasalsoloides Ostenf. [\u2261 Limbardasalsoloides Ikonn.] Ixerischinensis(Thunb.)Kitagawasubsp.chinensis s.l. Jacobaeaambracea (Turcz.) B.Nord. [= Senecioambraceus Turcz.] Jacobaeacannabifolia (Less.) E.Wiebe [\u2261 Seneciocannabifolius Less.] Jacobaeaerucifoliasubsp.argunensis (Turcz.) Veldkamp [\u2261 Senecioargunensis Turcz.] Jacobaeaerucifolia (L.) G.Gaertn., B.Mey. & Scherb. subsp.erucifolia [= Senecioerucifolius L.] Jacobaeavulgaris Gaertn. [\u2261 Seneciojacobaea L.] Jurineachaetocarpa (Ledeb.) Ledeb. Jurineamargalensis Iljin SEJurineamongolica Maxim. [= Jurineapotaninii Ilijn] [10\u201314]Jurineamultiflora B.Fedtsch. Kareliniacaspia Less. Kaschgariakomarovii (Krasch. & Rubtzov) Poljakov [\u2261 Tanacetumkomarovii Krasch. & Rubtzov] Klaseacardunculus Holub [\u2261 Serratulacardunculus Schischk.] Klaseacentauroides (L.) Cass. [\u2261 Serratulacentauroides L.] Klaseamarginata (Tausch) Kitag. [\u2261 Serratulamarginata Tausch] Klaseasogdiana (Bunge) L.Martins [6]Lactucaserriola L. [= Lactucasativasubsp.serriola (L.) Frietema] Lactucasibirica Benth. Lactucatatarica C.A.Mey. Lactucaundulata Ledeb. Leibnitziaanandria (L.) Turcz. Leontopodiumcampestre Hand.-Mazz. Leontopodiumconglobatum Hand.-Mazz. Leontopodiumleontopodioides (Willd.) Beauverd Leontopodiumnanum (Hook.f. & Thomson) Hand.-Mazz. [16]Leontopodiumochroleucum Beauverd Leontopodiumpalibinianum Beauverd Leuzeacarthamoides DC. [= Rhaponticumcarthamoides (Willd.) Iljin] [7]Leuzearepens (L.) D.J.N.Hind, [\u2261 Rhaponticumrepens (L.) Hidalgo \u2261 Acroptilonrepens (L.) DC.] Leuzeauniflora (L.) Holub [= Rhaponticumuniflorum (L.) DC.] Ligulariaaltaica DC. Ligulariafischerii (Ledeb.) Turcz. Ligulariaglauca (L.) O.Hoffm. [7]Ligulariahodgsonii Hook.f. Ligulariamongolica DC. Ligulariaprzewalskii Diels Ligulariasagitta (Maxim.) Mattf. [\u2261 Seneciosagitta Maxim.] Ligulariasibirica Cass. Lipschitziadivaricata (Turcz.) Zaika, Sukhor. & N.Kilian [\u2261 Scorzoneradivaricata Turcz.] Matricariachamomilla L. [= Matricariarecutita L.] [2]Neopallasiapectinata Poljakov SEOlgaealeucophylla (Turcz.) Iljin SEOlgaealomonossowii (Trautv.) Iljin [9]Omalothecasupina (L.) DC. [= Gnaphaliumsupinum L.] Packeracymbalaria (Pursh) W.A.Weber & \u00c1.L\u00f6ve [\u2261 Seneciocymbalaria Pursh] Paraseneciohastatus (L.) H.Koyama [\u2261 Cacaliahastata L.] Pentanemaasperum (Poir.) G.V.Boiko & Korniy. [\u2261 Inulaaspera Poir.] Pentanemabritannica (L.) D.Gut.Larr. [\u2261 Inulabritannica L.] Pentanemasalicinum (L.) D.Gut.Larr. [\u2261 Inulasalicina L.] Petasitesfrigidus (L.) Fr. [1]Petasitesradiatus (J.F.Gmel.) Toman [1]Petasitesrubellus (J.F.Gmel.) Toman Phalacrachenacalva (Ledeb.) Iljin [10]Picrisdavurica Fisch. Picrishieracioides L. Picrisjaponica Thunb. Piloselladublitzkii (B.Fedtsch. & Nevski) Sennikov [\u2261 Hieraciumdublitzkii B.Fedtsch. & Nevski] [7]Pilosellaechioides (L.) F.W.Schultz & Sch.Bip. [\u2261 Hieraciumechioides L.] Pulicariavulgaris Gaertn. [10]Rhinactinidiaeremophila (Bunge) Novopokr. [= Rhinactinidiaeremophilasubsp.grubovii Botsch.] Richteriapyrethroides Kar. & Kir. [\u2261 Pyrethrumpyrethroides (Kar. & Kir.) B.Fedtsch.] [7]Saussureaacuminata Turcz. SESaussureaalaschanica Maxim. Saussureaalata DC. Saussureaalpina (L.) DC. Saussureaamara (L.) DC. SESaussureaarctecapitulata Lipsch. Saussureabaicalensis B.L.Rob. SESaussureabogedaensis Yu J.Wang & J.Chen [14]SESaussureacatharinae Lipsch. [15]SESaussureaceterachifolia Lipsch. Saussureacongesta Turcz. [1]Saussureacontroversa DC. Saussureacoronata Schrenk [= Saussureadshungarica Iljin] [7]Saussureadaurica Adams SESaussureadorogostaiskii Palib. Saussureaelata Ledeb. [7]Saussureaelegans Ledeb. [= Saussureaamoena Kar. & Kir.] SESaussureaelongata DC. Saussureafoliosa Ledeb. Saussureaglacialis Herder SESaussureagrubovii Lipsch. Saussureagubanovii Kamelin [15]E Saussureainvolucrata (Kar. & Kit.) Sch.Bip. Saussureajaponica (Thunb.) DC. [9]Saussureaklementzii Lipsch. [7]SESaussureakrasnoborovii S.V.Smirn. [1]Saussureakrylovii Schischk. & Serg. [7]Saussurealaciniata Ledeb. Saussurealatifolia Ledeb. Saussurealeucophylla Schrenk SESaussurealipschitzii Filatova Saussureamongolica (Franch.) Franch. [5]Saussureaneoserrata Nakai Saussureaodontolepis Sch.Bip. [5]Saussureaodorata E.Pjak [7]E Saussureaorgaadayi Khanm. & Krasnob. Saussureaparviflora (Poir.) DC. SESaussureapopovii Lipsch. [14]Saussureapricei N.D.Simpson Saussureapseudoalpina N.D.Simpson Saussureapseudosalsa Lipsch. Saussureapulchella Fisch. SESaussureapurpurata (Fisch.) Lipsch. Saussurearamosa Lipsch. E Saussurearecurvata (Maxim.) Lipsch. Saussurearuncinata DC. Saussureasaichanensis Kom. E Saussureasalicifolia DC. [2\u20139]Saussureasalsa Spreng. Saussureaschanginiana (Wydler) Fisch. SESaussureasquarrosa Turcz. [1]Saussureastubendorffii Herder Saussureasubacaulis (Ledeb.) Serg. SESaussureasukaczevii Lipsch. Saussureaussuriensis Maxim. [5]Scorzoneraalbicaulis Bunge Scorzoneracurvata (Popl.) Lipsch. Scorzoneragrubovii Lipsch. E Scorzoneraparviflora Jacq. [14]Scorzoneraradiata Fisch. Scorzonerasinensis (Lipsch. & Krasch.) Nakai [9]Seneciodubitabilis C.Jeffrey & Y.L.Chen [\u2261 Seneciodubius Ledeb. nom. illegit. non Beck] Seneciokenteicus Grubov [2]E Senecionemorensis L. Seneciosubdentatus Ledeb. Seneciovulgaris L. Serratulacoronata L. Serratulakirghisorum Iljin [7]Solidagodahurica (Kitag.) Kitag. Solidagovirgaurea L. [7]Sonchelladentata (Ledeb.) Sennikov [\u2261 Sonchusdentatus Ledeb.] Sonchellastenoma (Turcz.) Sennikov [\u2261 Crepisstenoma Turcz.] [8\u201315]Sonchusarvensis L. Sonchusbrachyotus DC. Sonchusuliginosus M.Bieb. Stilpnolepisintricata (Franch.) C.Shih Symphyotrichumciliatum (Ledeb.) G.L.Nesom Synurusdeltoides (Aiton) Nakai Takhtajanianthaaustriaca (Willd.) Zaika, Sukhor. & N.Kilian [\u2261 Scorzoneraaustriaca Willd.] Takhtajanianthacapito (Maxim.) Zaika, Sukhor. & N.Kilian [\u2261 Scorzoneracapito Maxim.] Takhtajanianthaikonnikovii (Krasch. & Lipsch.) Zaika, Sukhor. & N.Kilian [\u2261 Scorzoneraikonnikovii Lipsch. & Krasch.] Takhtajanianthamongolica (Maxim.) Zaika, Sukhor. & N.Kilian [\u2261 Scorzoneramongolica Maxim.] [10\u201316]Takhtajanianthapseudodivaricata (Lipsch.) Zaika, Sukhor. & N.Kilian [\u2261 Scorzonerapseudodivaricata Lipsch] Takhtajanianthapusilla Nazarova [\u2261 Scorzonerapusilla Pall.] Takhtajanianthasubacaulis (Regel) Zaika, Sukhor. & N.Kilian [= Scorzonerasubacaulis (Regel) Lipsch.] [6]Tanacetumalatavicum Herder [\u2261 Pyrethrumalatavicum O.Fedtsch. & B.Fedtsch.] [7]Tanacetumchangaicum (Krasch.) K.Bremer & Humphries [\u2261 Pyrethrumchangaicum Krasch.] E Tanacetumcrassipes (Stschegl.) Tzvelev [7]Tanacetumkrylovianum (Krasch.) K.Bremer & Humphries [\u2261 Pyrethrumkrylovianum Krasch.] [7]Tanacetumlanuginosum Sch. [\u2261 Pyrethrumlanuginosum (Sch.Bip. & Herder) Tzvelev] SETanacetumpulchellum Sch. [\u2261 Pyrethrumpulchellum Turcz.] [7]Tanacetumpulchrum Sch. [= Pyrethrumpulchrum Ledeb.] Tanacetumtanacetoides (DC.) Tzvelev Tanacetumvulgare L. [= Tanacetumboreale Fisch. & DC.] Taraxacumarmeriifolium Soest Taraxacumasiaticum Dahlst. Taraxacumatrans Schischk. Taraxacumbessarabicum (Hornem.) Hand.-Mazz. Taraxacumbicorne Dahlst. SETaraxacumbornuurense R.Doll Taraxacumbrevirostre Hand.-Mazz. Taraxacumceratophorum (Ledeb.) DC. [= Taraxacumaltaicum Schischk.] Taraxacumcollinum DC. Taraxacumdealbatum Hand.-Mazz. Taraxacumdissectum Ledeb. Taraxacumeriopodum DC. Taraxacumerythrospermum Andrz. [3]Taraxacumglabrum DC. Taraxacumglaucanthum Nakai Taraxacumgoloskokovii Schischk. Taraxacuminimitabile Kirschner & \u0160t\u011bp\u00e1nek [13]E Taraxacumjunatovii Tzvelev E Taraxacumkok-saghyz Rodin SETaraxacumkrasnoborovii Krasnikov [7]SETaraxacumkrylovii Krasnikov & Khanm. [7]Taraxacumleucanthum Ledeb. Taraxacumlinczevskyi Schischk. [7]SETaraxacumlongicorne Dahlst. Taraxacumluridum G.E.Haglund Taraxacumlyratum (Ledeb.) DC. Taraxacummacilentum Dahlst. Taraxacummicrospermum Schischk. [= Taraxacumcompactum Schischk.] Taraxacumminutilobum Popov [7]Taraxacummongolicum Hand.-Mazz. Taraxacummongoliforme R.Doll Taraxacummonochlamydeum Hand.-Mazz. Taraxacummujense Petrochenko Taraxacummultisectum Kitag. [9]Taraxacumofficinale F.H.Wigg. Taraxacumparvulum DC. [14]Taraxacumpawlodarskum R.Doll [= Taraxacumustamenum R.Doll] [7]Taraxacumpingue Schischk. Taraxacumpseudoatratum Orazova [6]SETaraxacumpseudonivale Malyschev [1]Taraxacumpuberulum G.E.Haglund [14]SETaraxacumsangilense Krasnob. & Khanm. Taraxacumscariosum (Tausch) Kirschner & \u0160t\u011bp\u00e1nek Taraxacumselengensis Tzvelev [3]E Taraxacumsinicum Kitag. [= Taraxacumborealisinense Kitam.] [3\u201316]SETaraxacumsmirnovii M.S.Ivanova [7]SETaraxacumsongoricum Schischk. Taraxacumstanjukoviczii Schischk. Taraxacumsubmacilentum Tzvelev [7]E Taraxacumsumneviczii Schischk. Taraxacumtibetanum Hand.-Mazz. Taraxacumturgaicum Schischk. SETaraxacumtuvense Krasnob. & Krasnikov [1]Tephroserisflammea (DC.) Holub [\u2261 Senecioflammeus DC.] [5]Tephroserisintegrifoliasubsp.atropurpurea (Ledeb.) B.Nord. [1]Tephroserisintegrifolia(L.)Holubsubsp.integrifolia [= Seneciocampestris (Retz.) DC.] Tephroseriskirilowii (DC.) Holub [5]Tephroserispalustris (L.) Rchb. [\u2261 Seneciopalustris (L.) Hook.] SETephroserisporphyrantha (Schischk.) Holub [= Senecioporphyranthus Schischk.] Tephroserispraticola (Sisk. & Serg.) Holub [= Senecioasiatica Schischk. & Serg.] Tephroserispricei (N.D.Simpson) Holub [\u2261 Seneciopricei N.D.Simpson] SETephroserissukaczevii (Schischk.) Holub [\u2261 Seneciosukaczevii Schischk.] Tephroseristurczaninovii (DC.) Holub [\u2261Seneciosumneviczii Schischk. & Serg.] Tephroserisvereszczaginii (Schischk. & Serg.) Holub [\u2261 Senecioveresczaginii Schischk. & Serg.] [7]Tibetiodesflaccida (Bunge) G.L.Nesom [\u2261 Erigeronflaccidus (Bunge) Botsch.] Tragopogonkasahstanicus S.A.Nikitin [7]Tragopogonorientalis L. Tragopogonruber S.G.Gmel. Tragopogonsongoricus S.A.Nikitin SETragopogontrachycarpus S.A.Nikitin Tripleurospermumambiguum (Ledeb.) Franch. & Sav. [\u2261 Matricariaambigua (Ledeb.) Krylov] Tripoliumpannonicum (Jacq.) Dobrocz. [\u2261 Tripoliumpannonicum Jacq.] Trommsdorffiaciliata (Thunb.) Soj\u00e1k [\u2261 Hypochaerisciliata (Thunb.) Makino] [5]SETugarinoviamongolica Iljin Turczaninoviafastigiata (Fisch.) DC. [\u2261Asterfastigiatus Fisch.] Vickifunkiasongarica (Fisch.) C.Ren [\u2261 Ligulariasongarica (Fisch.) Y.Ling] [14]Vickifunkiathomsonii (C.B.Clarke) C.Ren [\u2261 Ligulariathomsonii (C.B.Clarke) Pojark.] [14]Vickifunkiathyrsoidea (Ledeb.) C.Ren [\u2261 Ligulariathyrsoidea (Ledeb.) DC.] Balsaminaceae29. A.Rich. (1 genus and 2 species)Impatiensnoli-tangere L. Impatiensparviflora DC. [7]Berberidaceae30. Juss. (1 genus and 2 species)Berberisamurensis Rupr. [5]Berberissibirica Pall. Betulaceae31. Gray (2 genera and 9 taxa)Alnusalnobetulasubsp.fruticosa (Rupr.) Raus Betulafruticosa Pall. Betulamandshuricasubsp.tauschii (Regel) Kamelin Betulamicrophylla Bunge Betulananasubsp.exilis (Sukachev) Hult\u00e9n Betulananasubsp.rotundifolia (Spach) Malyschev Betulaovalifolia Rupr. Betulapendulasubsp.mandshurica (Regel) Ashburner & McAll. BetulapendulaRothsubsp.pendula Biebersteiniaceae32. Schnizl. (1 genus and 1 species)Biebersteiniaodora Stephan Bignoniaceae33. Juss. (1 genus and 1 species)SEIncarvilleapotaninii Batalin Boraginaceae34. Juss. (24 genera and 78 taxa)Craniospermum Lehm. have been described from Mongolia by Ovczinnikova and Korolyuk (2016) and Arnebiatibetica previously known as a synonym of A.guttata, differs from A.guttata based on floral morphology and plastid genome characteristics discovered by Note: Since Amblynotusrupestris Popov [\u2261 Eritrichiumrupestre (Georgi) Bunge] Anchusaarvensis (L.) M.Bieb. SEAnoplocaryumcompressum Ledeb. [\u2261 Echinospermumcompressum (Ledeb.) Turcz.] Anoplocaryumtenellum A.L.Ebel & Rudaya [\u2261 Microulatenella (A.L.Ebel & Rudaya)] [7]E SEAnoplocaryumturczaninovii Krasnob. Arnebiadecumbens Coss. & Kralik Arnebiafimbriata Maxim. Arnebiaguttata Bunge Arnebiatibetana Kurz [7]Asperugoprocumbens L. Asperulagobicola Grubov [= Asperulasaxicola Grubov] E SECraniospermumcanescens DC. Craniospermumdesertorum Ovczinnikova & A.Korolyuk [7]E Craniospermumgubanovii Ovczinnikova [14]E Craniospermumkamelinii Ovczinnikova [7]E SECraniospermummongolicum I.M.Johnst. Craniospermummontanostepposum Ovczinnikova [7]E Craniospermumpseudotuvinicum Ovczinnikova & A.Korolyuk [10]E SECraniospermumtuvinicum Ovczinnikova Craniospermumvolkovae Ovczinnikova [10]E Cynoglossumdivaricatum Steph. SEEritrichiumalpinum Ovczinnikova [6]Eritrichiumpauciflorum DC. Eritrichiumpectinatum DC. [3]SEEritrichiumpulviniforme Popov SEEritrichiumsajanense Sipliv. [1]Eritrichiumthymifolium (DC.) Y.S.Lian & J.Q.Wang Eritrichiumtianschanicum Iljin [6]Eritrichiumvillosum (Ledeb.) Bunge [\u2261 Myosotisvillosa Ledeb.] Hackeliadeflexa (Wahlenb.) Opiz [\u2261 Myosotisdeflexa Wahlenb.] Heliotropiumellipticum Ledeb. Lappulabalchaschensis Popov Lappulabrachycentroides Popov [3]Lappulaconsanguinea G\u00fcrke Lappulacoronifera Popov [3]Lappuladuplicicarpa Pavlov SELappulagranulata (Krylov) Popov Lappulaheteracantha (Ledeb.) G\u00fcrke [7]Lappulaintermedia (Ledeb.) Popov Lappulakrylovii Ovczinnikova, Pjak & A.L.Ebel [7]Lappulamacrantha (Ledeb.) G\u00fcrke Lappulamicrocarpa G\u00fcrke Lappulamyosotis Wolf Lappulapatula Asch. Lappularedowskii (Hornem.) Greene Lappulasemiglabra (Ledeb.) G\u00fcrke Lappulastricta (Ledeb.) G\u00fcrke Lappulatadshikorum Popov [7]Lappulatenuis G\u00fcrke Lappulatianschanica Popov & Zakirov [7]Lappulatuvinica Ovczinnikova [6]Lindelofiastylosa (Kar. & Kir.) Brand [\u2261 Cynoglossumstylosum Kar. & Kir.] Mertensiadavurica (Sims) G.Don [= Mertensiaochroleuca Ikonn.-Gal.] Mertensiapallasii G.Don [7]Mertensiastylosa DC. Mertensiatarbagataica B.Fedtsch. [7]SEMicroulatibeticavar.pratensis (Maxim.) W.T.Wang [\u2261 Tretocaryapratensis Maxim.] Myosotisalpestris F.W.Schmidt Myosotisaustrosibirica O.D.Nikif. Myosotisbaltica Sam. Myosotiscaespitosa Schultz Myosotiskrylovii Serg. Myosotisscorpioides L. [2]Myosotisstricta Link [7]Noneacaspica G.Don Noneapulla DC. Nonearossica Steven [3]Onosmafuyunensis Y.He & Q.R.Liu [7]Onosmagmelinii Ledeb. OnosmasetosaLedeb.subsp.setosa [7]Onosmasetosasubsp.transrhymnensis (Klokov) Kamelin Pseudolappulaoccultata (Popov) Q.R.Liu & D.H.Liu [\u2261 Lappulaoccultata Popov] [14]Pulmonariadacica (Simonk.) Simonk. [= Pulmonariamollissima A.Kern.] Rinderatetraspis Pall. [14]Rocheliabungei Trautv. Rochelialeiocarpa Ledeb. Stenosoleniumsaxatile Turcz. [\u2261 Anchusasaxatilis Pall.] Tournefortiasibirica L. [= Messerschmidiasibirica (L.) L.] Brassicaceae35. Burnett (51 genera and 138 taxa)Brassicaceae was recently revised by Lepidiumgobicum V.I.Dorof. was newly described from Mongolia and China by Lepidiumapetalum Willd. Heynh. Arabisborealis Andrz. Barbareaorthoceras Ledeb. Barbareavulgaris W.T.Aiton Brayahumilis (C.A.Mey.) B.L.Rob. Brayarosea Bunge Brayasiliquosa Bunge [1]Camelinamicrocarpa Andrz. Capsellabursa-pastoris (L.) Medik. Capsellaorientalis Klokov [\u2261 Capsellabursa-pastorissubsp.orientalis (Klokov) Tzvelev] Cardaminebellidifolia L. Cardamineimpatiens L. [6]Cardamineleucantha (Tausch) O.E.Schulz [5]Cardaminemacrophylla Willd. Cardamineparviflora L. Cardaminepratensis L. Cardamineprorepens Fisch. [5]Cardaminetrifida (Lam.) B.M.G.Jones [5]Catolobuspendulus (L.) Al-Shehbaz [= Arabispendula L.] Chorisporabungeana Fisch. & C.A.Mey. [7]Chorisporasibirica (L.) DC. Chorisporatenella DC. Clausiaaprica Trotzky Clausiatrichosepala (Turcz.) F.Dvo\u0159\u00e1k [4]Crucihimalayamollissima (C.A.Mey.) Al-Shehbaz [\u2261 Sisymbriummollissimum C.A.Mey.] SECrucihimalayarupicola (Krylov) A.L.Ebel & D.A.German [\u2261 Arabisrupicola Krylov] Dendroarabisfruticulosa (C.A.Mey.) D.A.German & Al-Shehbaz [\u2261 Arabisfruticulosa C.A.Mey.] Descurainiasophia (L.) Webb SEDontostemoncrassifolius (Bunge) Maxim. Dontostemondentatus Ledeb. [5]SEDontostemonelegans Maxim. Dontostemongubanovii (D.A.German) D.A.German [\u2261 Dontostemonsenilissubsp.gubanovii D.A.German] E Dontostemonintegrifolius (L.) Ledeb. Dontostemonmicranthus C.A.Mey. SEDontostemonperennis C.A.Mey. Dontostemonpinnatifidus (Willd.) Al-Shehbaz & H.Ohba [\u2261 Cheiranthuspinnatifidus Willd.] SEDontostemonsenilis Maxim. Drabaalpina L. [6]Drabaaltaica (C.A.Mey.) Bunge SEDrababaicalensis Tolm. Drabaeriopoda Turcz. Drabafladnizensis Wulfen Drabahirta L. Drabakusnetzovii (Turcz.) Hayek Drabalanceolata Royle Drabamongolica Turcz. Drabanemorosa L. Drabaochroleuca Bunge Drabaoreades Schrenk SEDrabapygmaea Turcz. Drabasibirica Thell. Drabastenocarpa Hook.f. & Thomson [7]Drabasubamplexicaulis C.A.Mey. Drabaturczaninowii Pohle Erysimumandrzejowskianum Bess. [7]Erysimumcheiranthoidessubsp.altum Ahti [without indication of regions]ErysimumcheiranthoidesL.subsp.cheiranthoides Erysimumcheiranthoidessubsp.transiliense (Popov) D.A.German [\u2261 Erysimumtransiliense Popov] [7]Erysimumflavum(Georgi)Bobrovsubsp.flavum Erysimumflavumsubsp.altaicum (C.A.Mey.) Polozhij SEErysimumkotuchovii D.A.German [7]Erysimumledebourii D.A.German [7]Erysimummarschallianum Andrz. SEErysimummongolicum D.A.German Erysimumsisymbrioides C.A.Mey. Eutremaedwardsiisubsp.compactum (O.E.Schulz) A.L.Ebel [\u2261 Eutremacompactum O.E.Schulz] [7]EutremaedwardsiiR.Br.subsp.edwardsii Eutremasalsugineum Al-Shehbaz & Warwick [\u2261 Sisymbriumsalsugineum Pall.] Galitzkyamacrocarpa Botsch. [\u2261 Berteroamacrocarpa Ikonn.-Gal.] E SEGalitzkyapotaninii (Maxim.)Botsch. SEGoldbachiaikonnikovii Vassilcz. GoldbachiapendulaBotsch. Hesperissibirica L. Hornungiaprocumbens Hayek Iljinskaeaplanisiliqua (Fisch. & C.A.Mey.) Al-Shehbaz [\u2261 Conringiaplanisiliqua Fisch. & C.A.Mey.] Isatiscostata C.A.Mey. Isatisgymnocarpa (Fisch.) Al-Shehbaz, Moazzeni & Mumm. [\u2261 Tauscheriagymnocarpa Fisch.] [14]Isatismulticaulis (Kar. & Kir.) Jafri [14]Isatisoblongata DC. Leiocarpaeacochlearioides (Murray) D.A.German & Al-Shehbaz [\u2261 Buniascochlearioides Murray] [1]Leiosporaexscapa (C.A.Mey.) F.Dvo\u0159\u00e1k [\u2261 Parryaexscapa C.A.Mey.] Lepidiumaffine Ledeb. [\u2261 Lepidiumlatifoliumsubsp.affine (Ledeb.) Kitag.] Lepidiumamplexicaule Willd. Lepidiumapetalum Willd. Lepidiumappelianum Al-Shehbaz Lepidiumcartilagineum Thell. Lepidiumcordatum Willd. Lepidiumlacerum C.A.Mey. [= Lepidiumsongaricum Schrenk] Lepidiumobtusum Basiner Litwinowiatenuissima Woronow [14]Macropodiumnivale R.Br. Matthiolasuperba Conti [14]Megacarpaeamegalocarpa Schischk. [14]Meniocuslinifolius (Willd.) DC. [\u2261 Alyssumlinifolium Willd.] SEMicrostigmabrachycarpum Botsch. SEMicrostigmadeflexum (Bunge) Juz. Neotorulariabrevipes (Kar. & Kir.) Hedge & J.L\u00e9onard [\u2261 Sisymbriumbrevipes F.Muell.] Noccaeaferganensis (N.Busch) Czerep. [\u2261 Thlaspiferganense N.Busch] [7]Noccaeathlaspidioides F.K.Mey. [\u2261 Lepidiumthlaspidioides Pall. = Thlaspicochleariforme DC.] Odontarrhenaobovata C.A.Mey. [\u2261 Alyssumobovatum (C.A.Mey.) Turcz.] [1\u201310]Olimarabidopsispumila (Stephan) Al-Shehbaz [\u2261 Sisymbriumpumilum Stephan] [14]SEPachyneurumgrandiflorum Bunge Pugioniumdolabratum Maxim. SEPugioniumpterocarpum Kom. [10]Rhammatophyllumerysimoides (Kar. & Kir.) Al-Shehbaz & O.Appel [\u2261 Arabiserysimoides Kar. & Kir.] Rorippabarbareifolia (DC.) Kitag. [2]Rorippadogadovae Tzvelev Rorippapalustris Besser Sisymbriumbrassiciforme C.A.Mey. Sisymbriumheteromallum C.A.Mey. Sisymbriumloeselii L. Sisymbriumpolymorphum (Murr.) Roth Sisymbriumsubspinescens Bunge [14]Smelowskiaalba B.Fedtsch. SESmelowskiaaltaica (Pobed.) Botsch. Smelowskiabifurcata (Ledeb.) Botsch. Smelowskiacalycina (Stephan) C.A.Mey. [= Lepidiumcalycinum Steph.] SESmelowskiacalycinasubsp.pectinata (Bunge) D.A.German [= Hutchinsiapectinata Bunge] Smelowskiamongolica Kom. [3]E SESterigmostemumviolaceum (Botsch.) H.L.Yang Steveniaalyssoides Adams & Fisch. SESteveniaalyssoidessubsp.zinaidae Kamelin [\u2261 Steveniazinaidae Malyschev] Steveniacanescens (DC.) D.A.German [\u2261 Alyssumcanescens DC. \u2261 Ptilotrichumcanescens (DC.) C.A.Mey.] SESteveniacheiranthoidesDC.subsp.cheiranthoides Steveniacheiranthoidessubsp.incarnata (Kamelin) D.A.German SESteveniadahurica (Peschkova) D.A.German & Al-Shehbaz SESteveniasergievskajae (Krasnob.) Kamelin & Gubanov [\u2261 Alyssumsergievskajae Krasnob.] [3]SESteveniatenuifolia (Stephan) D.A.German [\u2261 Alyssumtenuifolium Steph.] Strigosellaafricana (L.) Botsch. Strigosellabrevipes (Bunge) Botsch. [14]Subulariaaquatica L. Tetracmequadricornis (Steph.) Bunge Thlaspiarvense L. Thlaspiceratocarpum Murray [\u2261 Carpocerasceratocarpum N. Busch] Turritisglabra L. [7]Butomaceae36. Mirb. (1 genus and 2 species)Butomusjunceus Turcz. Butomusumbellatus L. Campanulaceae37. Juss. (4 genera and 18 taxa)Adenophorachangaica Gubanov & Kamelin [3]E Adenophoragmelinii Fisch. Adenophoralamarkii Fisch. [\u2261 Campanulalamarckii D.Dietr.] Adenophoraliliifolia (L.) A.DC. [\u2261 Campanulaliliifolia L.] Adenophorapereskiifolia (Fisch.) G.Don Adenophorastenanthina (Ledeb.) Kitagawa [= Adenophoracrispata Turcz.] Adenophoratricuspidata A.DC. Adenophoratriphylla (Thunb.) A.DC. [\u2261 Campanulatriphylla Thunb.] Campanulacervicaria L. [2]Campanuladasyantha M.Bieb. Campanulaglomerata L. Campanulapunctata Lam. [9]Campanularotundifolia L. [6]Campanulasteveniisubsp.altaica (Ledeb.) Fed. [\u2261 Campanulaaltaica Ledeb.] [7]Campanulasteveniisubsp.turczaninovii (Fed.) Victorov [\u2261 Campanulaturczaninovii Fed.] Campanulasteveniisubsp.wolgensis (P.A.Smirn.) Fed. [\u2261 Campanulawolgensis P.A.Smirn.] [7]Codonopsisclematidea C.B.Clarke [7]Platycodongrandiflorus A.DC. [5]Caprifoliaceae38. Juss. (5 genera and 24 taxa)Linnaeaborealis L. Loniceracaeruleasubsp.altaica Gladkova [\u2261 Loniceraaltaica Pall.] LoniceracaeruleaL.subsp.caerulea [6]Loniceracaeruleavar.venulosa (Maxim.) Vorosch. [\u2261 Loniceravenulosa Maxim.] [5]Lonicerachrysantha Turcz. [5]Lonicerahispida Pall. Loniceramicrophylla Willd. [\u2261 Caprifoliummicrophyllum (Willd.) Kuntze] Loniceratatarica L. [4]Patriniaheterophylla Bunge [9]Patriniaintermedia Roem. & Schult. Patriniarupestris Dufr. [\u2261 Valerianarupestris Pall.] Patriniascabiosifolia Fisch. Patriniasibirica (L.) Juss. [1\u20137]Scabiosacomosa Fisch. Scabiosaochroleuca L. Valerianaaltaica Sumnev. Valerianaalternifolia Ledeb. [= Valerianadahurica Sumnev.] Valerianacapitata Pall. [1]Valerianadubia Bunge Valerianamartjanovi Krylov [= Valerianasaichanensis Kom.] [13]Valerianaofficinalis L. Valerianapetrophila Bunge SEValerianatangutica Batalin [16]SEValerianatransjenisensis Kreyer Caryophyllaceae39. Juss. (20 genera and 97 taxa)Acanthophyllumpungens Boiss. Arenarialeptoclados Guss. [7]Arenariaserpyllifolia L. Cerastiumalpinum L. [6]Cerastiumarvense L. Cerastiumcerastoides (L.) Britton [\u2261 Dichodoncerastoides (L.) Rchb. \u2261 Stellariacerastoides L.] Cerastiumdavuricum Fisch. Cerastiumfalcatum (Gren.) Bunge [\u2261 Stellariafalcata Ser.] [14]Cerastiumholosteoides Fr. [\u2261 Cerastiumfontanumf.holosteoides (Fr.) M.B.Wyse Jacks.] [2]Cerastiumlithospermifolium Fisch. Cerastiummaximum L. [13]Cerastiumpauciflorum Steven Cerastiumpusillum Ser. Cherleriaarctica (Steven) A.J.Moore & Dillenb. [\u2261 Minuartiaarctica (Steven) Graebn.] Cherleriabiflora (L.) A.J.Moore & Dillenb. [\u2261 Minuartiabiflora (L.) Schinz & Thell.] Dianthuschinensis L. [= Dianthusversicolor Fisch.] Dianthuscrinitussubsp.soongoricus (Schischk.) Kozhevn. [\u2261 Dianthussoongoricus Schischk.] Dianthusramosissimus Pall. [10]Dianthusrepens Willd. [\u2261 Dianthuschinensissubsp.repens (Willd.) Vorosch.] Dianthussuperbus L. Eremogoneandrosacea (Grubov) Ikonn. [\u2261 Arenariaandrosacea Grubov] [13]Eremogoneasiatica (Schischk.) Ikonn. [\u2261 Arenariaasiatica Schischk.] [7]Eremogonecapillaris (Poir.) Fenzl [\u2261 Arenariacapillaris Poir.] Eremogonejuncea (M.Bieb.) Fenzl [\u2261 Arenariajuncea M.Bieb.] Eremogonemeyeri (Fenzl) Ikonn. [\u2261 Arenariameyeri Fenzl] SEEremogonemongolica (Schischk.) Ikonn. [\u2261 Arenariamongolica Schischk.] [7]SEGymnocarposprzewalskii Maxim. [\u2261 Paronychiaprzewalskii (Bunge) Rohweder & Urmi-K\u00f6nig] Gypsophilaaltissima L. [7]Gypsophilacapituliflora Rupr. Gypsophilacephalotes (Schrenk) F.N.Williams Gypsophiladavurica Fenzl [\u2261 Gypsophilapatriniisubsp.davurica (Fenzl) Kozhevn.] Gypsophilapaniculata L. Gypsophilapatrinii Ser. Gypsophilaperfoliata L. [10]Gypsophilasericea (Ser.) Krylov [\u2261 Arenariasericea Ser.] [7]Herniariacaucasica Rupr. [7]Herniariaglabra L. [7]Heterochroadesertorum Bunge [\u2261 Gypsophiladesertorum Fenzl] Lepyrodiclisholosteoides (C.A.Mey.) Fenzl [\u2261 Gouffeiaholosteoides C.A.Mey.] Moehringialateriflora (L.) Fenzl [\u2261 Arenarialateriflora L.] Moehringiaumbrosa (Bunge) Fenzl [\u2261 Arenariaumbrosa Bunge] Pseudocherlerialaricina (L.) Dillenb. & Kadereit [\u2261 Minuartialaricina Mattf.] Pseudostellariarupestris (Turcz.) Pax Sabulinaregeliana (Trautv.) Dillenb. & Kadereit [= Minuartiaregeliana (Trautv.) Mattf.] [3]Sabulinastricta (Sw.) Rchb. [\u2261 Minuartiastricta (Sw.) Hiern] Sabulinaverna Rchb. [\u2261 Minuartiaverna (L.) Hiern] Saginasaginoides (L.) H.Karst. [7]Saponariafloribunda (Kar. & Kir.) Boiss. [\u2261 Psammophiliellafloribunda (Kar. & Kir.) Ikonn.] [14]Silenealexandrae B.Keller [14]Silenealtaica Pers. Sileneaprica Turcz. [\u2261 Ussuriaaprica (Turcz.) Tzvelev] Sileneborysthenica (Gruner) Walters Silenebungei Bocquet Silenechamarensis Turcz. [\u2261 Silenetenuissubsp.chamarensis (Turcz.) Kozhevn.] Sileneconoidea L. [7]Silenefoliosa Maxim. Silenegraminifolia Otth [= Silenesobolevskajae Czerep.] Silenegubanovii Lazkov SESileneintramongolica Lazkov Silenejeniseensis Willd. [= Sileneiche-bogdo Grubov] Silenelatifoliasubsp.alba (Mill.) Greuter & Burdet [\u2261 Lychnisalba Mill.] [7]Silenemongolica Maxim. E Silenequadriloba Turcz. Silenerepens Patrin Silenesamojedorum (Sambuk) Oxelman [\u2261 Lychnissibiricasubsp.samojedorum Sambuk.] Silenesibirica Pers. [14]Silenesongarica Bocquet [= Gastrolychnisbrachypetala (Hornem.) Tolm. & Kozhanczikov] Silenesuaveolens Kar. & Kir. Sileneuralensis (Rupr.) Bocquet Sileneviolascens (Tolm.) V.V.Petrovsky & Elven [\u2261 Gastrolychnisviolascens Tolm.] [7]Sileneviscosa Schleich. Silenevulgaris (Moench) Garcke Spergulariamarina (L.) Besser [\u2261 Arenariarubravar.marina L.] Spergulariasegetalis G.Don [14]Stellariaalsinoides Boiss. & Buhse Stellariaamblyosepala Schrenk Stellariabrachypetala Bunge Stellariabungeana Fenzl [\u2261 Hylebiabungeana (Fenzl) Tzvelev] Stellariacherleriae (Fisch.) F.N.Williams [\u2261 Arenariacherleriae Fisch.] Stellariacrassifolia Ehrh. Stellariadavurica Willd. Stellariadepressa Schmid [7]Stellariadichotoma L. [1\u201314]Stellariadichotomavar.lanceolata Bunge [= Stellariagypsophiloides Fenzl] Stellariadiscolor Turcz. Stellariafilicaulis Makino Stellariaimbricata Bunge Stellariairrigua Bunge Stellarialongifolia Muhl. Stellarialongipes Goldie [= Stellularialongipes (Goldie) MacMill.] Stellariamartjanovii Krylov [= Mesostemmamartjanovii (Krylov) Ikonn.] [7]Stellariamedia (L.) Vill. [= Alsinemedia L.] Stellariapalustris Ehrh. Stellariapetraea Bunge Stellariapulvinata Grubov Stellariaradians L. Stellariazolotuchinii A.L.Ebel [\u2261 Stellariaglandulifera N.Zolot. nom. illegit.] Celastraceae40. R.Br. (2 genera and 3 species)Euonymusmaackii Rupr. Parnassialaxmannii Pall. Parnassiapalustris L. Ceratophyllaceae41. Gray (1 genus and 2 taxa)Ceratophyllumdemersum L. Ceratophyllumplatyacanthumsubsp.oryzetorum (Kom.) Les [10]Cleomaceae42. Bercht. & J.Presl (1 genus and 1 species)Cleomegobica Grubov [15]E Convolvulaceae43. Juss. (4 genera and 15 species)Calystegiahederacea Wall. Calystegiapellita G.Don [= Calystegiadahurica Herb.] Calystegiasepium (L.) R.Br. [\u2261 Convolvulussepium L.] [14]Calystegiasubvolubilis G.Don Convolvulusammannii Desr. Convolvulusarvensis L. Convolvulusfruticosus Pall. Convolvulusgortschakovii Schrenk Convolvulustragacanthoides Turcz. Cuscutaaustralis R.Br. [9]Cuscutachinensis Lam. Cuscutaeuropaea L. Cuscutalupuliformis Krock. Cuscutamonogyna Vahl [\u2261 Monogynellamonogyna (Vahl) Hada\u010d] Merremiasibirica (L.) Hallier f. [3]Cornaceae44. Bercht. & J.Presl (1 genus and 1 species)Cornusalba L. Crassulaceae45. J.St.-Hil. (6 genera and 17 taxa)Crassulaaquatica (L.) Sch\u00f6nland Hylotelephiumewersii (Ledeb.) H.Ohba [\u2261 Sedumewersii Ledeb.] Hylotelephiumpallescens (Freyn) H.Ohba Hylotelephiumtelephium (L.) H.Ohba [\u2261 Sedumtelephium L.] [1\u201310]Orostachysfimbriata (Turcz.) A.Berger [\u2261 Cotyledonfimbriata Turcz.] Orostachysmalacophylla Fisch. [\u2261 Cotyledonmalacophylla Pall.] Orostachysspinosa (L.) Sweet [\u2261 Cotyledonspinosa L.] Orostachysthyrsiflora Fisch. Phedimusaizoon (L.) \u2018t Hart [\u2261 Sedumaizoon L.] [1\u201314]Phedimushybridus (L.) \u2018t Hart [\u2261 Sedumhybridium L.] Pseudosedumlievenii A.Berger Rhodiolaalgida (Ledeb.) Fisch. & C.A.Mey. Rhodiolacoccinea (Royle) Boriss. [7]Rhodiolalitwinowii Boriss. Rhodiolaquadrifida Fisch. & C.A.Mey. Rhodiolarosea L. [\u2261 Sedumroseum (L.) Scop.] Rhodiolastephani (Cham.) Trautv. & C.A.Mey. [= Rhodiolakrylovii Polozhij & Revjakina = Rhodiolapinnatifida Boriss. = Rhodiolasubpinnata (Krasnob.) Krasnob.] Cynomoriaceae46. Endl. (1 genus and 1 species)Cynomoriumsongaricum Rupr. [\u2261 Cynomoriumcoccineumsubsp.songaricum (Rupr.) J.L\u00e9onard] [10\u201316]Cyperaceae47. Juss. (10 genera and 131 taxa)Kobresia Willd. have been recorded in Mongolia Kukkonen Blysmusrufus Link Bolboschoenusmaritimus (L.) Palla [\u2261 Scirpusmaritimus L.] Bolboschoenusmaritimussubsp.affinis (Roth) T.Koyama [= Bolboschoenuspopovii T.V.Egorova] [10\u201316]Bolboschoenusplaniculmis (F.Schmidt) T.V.Egorova Carexaccrescens Ohwi [= Carexpallida C.A.Mey.] [2]Carexacuta L. Carexalatauensis S.R.Zhang [= Kobresiahumilis (C.A.Mey.) Serg.] Carexalba Scop. Carexaltaica (Gorodkov) V.I.Krecz. [\u2261 Carexorbicularissubsp.altaica (Gorodkov) T.V.Egorova] [3]Carexamgunensis F.Schmidt [= Carexchloroleuca Meinsh.] Carexappendiculata K\u00fck. Carexargunensis Turcz. [\u2261 Carexrupestrissubsp.argunensis (Turcz.) Vorosch.] Carexarnellii Christ Carexaterrima Hoppe Carexatherodes Spreng. Carexatrofusca Schkuhr Carexbigelowiisubsp.ensifolia (Gorodkov) Holub Carexbigelowiisubsp.rigidioides (Gorodkov) T.V.Egorova Carexbistaminata (W.Z.Di & M.J.Zhong) S.R.Zhang [\u2261 Kobresiabistaminata W.Z.Di & M.J.Zhong = Kobresiamyosuroides (Vill.) Fiori] Carexbohemica Schreb. [4]Carexborealipolaris S.R.Zhang [= Kobresiasibirica (Turcz.) Boeck. = Kobresiasmirnovii Ivanova] Carexbrunnescens (Pers.) Poir. [\u2261 Carexcurtavar.brunnescens Pers.] Carexcanescens L. Carexcapillifolia (Decne.) S.R.Zhang [= Kobresiacapilliformis Ivanova] Carexcapitata L. Carexcapricornis Meinsh. [10]Carexcaryophyllea Latourr. Carexcespitosa L. Carexchordorrhiza L.f. [2]Carexcoriophora Fisch. & C.A.Mey. Carexcuraica Kunth Carexdahurica K\u00fck. Carexdelicata C.B.Clarke Carexdiandra Schrank Carexdiluta M.Bieb. [10]Carexdistanssubsp.aspratilis (V.I.Krecz.) T.V.Egorova Carexduriuscula C.A.Mey. Carexeleusinoides Turcz. Carexenervis C.A.Mey. Carexeremopyroides V.I.Krecz. Carexericetorum Pollich [4]Carexglobularis L. Carexgotoi Ohwi [\u2261 Carexsongoricasubsp.gotoi (Ohwi) Popov] Carexhancockiana Maxim. Carexheterolepis Bunge [5]SECarexiljinii V.I.Krecz. Carexkaroi Freyn [= Carexselengensis N.A.Ivanova] Carexkorshinskii Kom. Carexlachenalii Schkuhr Carexlanceolata Boott [2]Carexlasiocarpa Ehrh. [2]Carexlaxa Wahlenb. Carexledebouriana C.A.Mey. [\u2261 Carexcapillarissubsp.ledebouriana (C.A.Mey.) Vorosch.] Carexleporina L. Carexlimosa L. [2]Carexlithophila Turcz. [\u2261 Carexdistichasubsp.lithophila (Turcz.) D.H\u00e4met-Ahti] Carexloliacea L. Carexmacrogyna Turcz. Carexmacroprophylla (Y.C.Yang) S.R.Zhang [\u2261 Kobresiafilifolia (Turcz.) C.B.Clarke \u2261 Kobresiafilifolia(Turcz.)C.B.Clarkevar.macroprophylla Y.C.Yang] CarexmagellanicaLam.subsp.irrigua (Wahlenb.) Hiitonen [\u2261 CarexlimosaL.var.irrigua Wahlenb.] Carexmedia R.Br. Carexmelanantha C.A.Mey. Carexmelanocephala Turcz. Carexmeyeriana Kunth Carexmicroglochin Wahlenb. Carexnigrasubsp.juncea (Fr.) So\u00f3 [= Carexjuncella T.M.Fries] Carexnorvegica Retz. Carexobtusata Lilj. Carexorbicularis Boott Carexpamirica(O.Fedtsch.)B.Fedtsch.subsp.dichroa (Freyn) T.V.Egorova [\u2261 CarexpullaGooden.subsp.dichroa Freyn] CarexparallelaLaest.subsp.redowskiana (C.A.Mey.) T.V.Egorova [\u2261 Carexredowskiana C.A.Mey.] Carexparva Nees [3]Carexpediformisvar.macroura (Meinsh.) K\u00fck. [\u2261 Carexmacroura Meinsh.] Carexpediformissubsp.pediformis Carexpraecox Schreb. Carexpseudofoetida K\u00fck. Carexpycnostachya Kar. & Kir. [\u2261 Carexcuraicasubsp.pycnostachya (Kar. & Kir.) T.V.Egorova] Carexraddei K\u00fck. [2]Carexrelaxa V.I.Krecz. Carexreptabunda (Trautv.) V.I.Krecz. Carexrhynchophysa Fisch. Carexrostrata Stokes Carexrupestris All. Carexsabulosa Turcz. Carexsabynensis Less. SECarexsajanensis V.I.Krecz. Carexsargentiana (Hemsl.) S.R.Zhang [\u2261 Kobresiasargentiana Hemsl. = Kobresiarobusta Maxim.] [3]Carexsaxatilis L. [= Carexsaxatilissubsp.laxa (Trautv.) Kalela] Carexschmidtii Meinsh. Carexsedakowii C.A.Mey. Carexsimpliciuscula Wahlenb. [= Kobresiasimpliciusculasubsp.subholarctica T.V.Egorova] Carexsongorica Kar. & Kir. Carexsordida Van Heurck & M\u00fcll.Arg. Carexstenophyllasubsp.stenophylloides (V.I.Krecz.) T.V.Egorova Carexsupermascula V.I.Krecz. Carextenuiflora Wahlenb. Carextomentosa L. Carextristissubsp.stenocarpa (Turcz.) T.V.Egorova Carexvaginatavar.petersii (C.A.Mey.) Akiyama [= Carexfalcata Turcz.] CarexvaginataTauschvar.vaginata Carexvesicata Meinsh. Carexwilliamsii Britton [2]Carexyamatsutana Ohwi [= Carexdiplasiocarpa V.I.Krecz.] Cyperusfuscus L. Cyperushamulosus M.Bieb. [\u2261 Mariscushamulosus (M.Bieb.) S.S.Hooper] [10]Cyperusmichelianus (L.) Delile [\u2261 Scirpusmichelianus L.] [4]Cyperuspannonicus Jacq. [\u2261 Juncelluspannonicus (Jacq.) C.B.Clarke] Eleocharisacicularis (L.) Roem. & Schult. Eleocharismamillata (H.Lindb.) H.Lindb. [10]Eleocharismitracarpa Steud. Eleocharispalustris (L.) Roem. & Schult. [\u2261 Scirpuspalustris L.] [1\u201316]Eleocharisquinqueflora (Hartmann) O.Schwarz [\u2261 Scirpusquinqueflorus Hartmann] Eleocharisuniglumis Schult. [= Eleocharisklingei (Meinsh.) B.Fedtsch.] Eleocharisyokoscensis (Franch. & Sav.) Tang & F.T.Wang [\u2261 Eleocharisacicularissubsp.yokoscensis (Franch. & Sav.) T.V.Egorova] Eriophorumaltaicum Meinsh. Eriophorumangustifolium Honck. Eriophorumangustifoliumsubsp.komarovii (V.N.Vassil.) M.S.Novos. [1\u201310]Eriophorumbrachyantherum Trautv. & C.A.Mey. Eriophorumcallitrix C.A.Mey. [1]Eriophorumchamissonis C.A.Mey. [= Eriophorummandshuricum Meinsh.] Eriophorumgracile W.D.J.Koch [= Eriophorumgracilesubsp.asiaticum (V.N.Vassil.) M.S.Novos.] Eriophorumhumile Turcz. Eriophorumvaginatum L. [3]Schoenoplectiellasupina (L.) Lye [= Schoenoplectussupinus (L.) Pall.] [10]Schoenoplectuslacustrissubsp.hippolytii (V.I.Krecz.) Kukkonen [\u2261 Scirpushippolyti V.I.Krecz.] Schoenoplectustabernaemontani (C.C.Gmel.) Pall. Schoenoplectustriqueter (L.) Palla [\u2261 Scirpustriqueter L.] [9]Scirpusorientalis Ohwi [\u2261 Scirpussylvaticussubsp.orientalis (Ohwi) Vorosch.] Scirpusradicans Schkuhr Trichophorumpumilum (Vahl) Schinz & Thell. Droseraceae48. Salisb. (2 genera and 3 species)Aldrovandavesiculosa L. [10]Droseraanglica Huds. [2]Droserarotundifolia L. [2]Elaeagnaceae49. Juss. (2 genera and 3 taxa)Elaeagnusangustifolia L. HippophaerhamnoidesSt.-Lag.subsp.mongolica Rousi [\u2261 Hippophaemongolica (Rousi) Tzvelev] HippophaerhamnoidesSt.-Lag.subsp.turkestanica Rousi [\u2261 Hippophaeturkestanica (Rousi) Tzvelev] [14]Ericaceae50. Durande (12 genera and 27 taxa)Arctostaphylosuva-ursi (L.) Spreng. [2]Arctousalpina (L.) Nied. [= Arbutusalpina L.] Cassiopeericoides D.Don [4]Chamaedaphnecalyculata (L.) Moench EmpetrumnigrumL.subsp.nigrum Empetrumnigrumsubsp.sibiricum (V.N.Vassil.) Kuvaev [\u2261 Empetrumsibiricum V.N.Vassil.] Monesesuniflora A.Gray Monotropahypopitys L. Orthiliaobtusata (Turcz.) H.Hara [\u2261 Pyrolasecundavar.obtusata Turcz.] Orthiliasecunda (L.) House [\u2261 Pyrolasecunda L] Phyllodocecaerulea (L.) Bab. [\u2261 Andromedacaerulea L.] [1]Pyrolaasarifoliasubsp.incarnata (DC.) A.E.Murray [\u2261 Pyrolarotundifoliavar.incarnata (DC.) A.P.Khokhr.] Pyrolachlorantha Sw. Pyroladaurica Kom. Pyrolamedia Sw. Pyrolaminor L. [2]Pyrolarotundifolia L. Rhododendronadamsii Rehder Rhododendronaureum Georgi Rhododendrondauricum L. Rhododendronlapponicum (L.) Wahlenb. Rhododendronledebourii Pojark. [\u2261 Rhododendrondauricumsubsp.ledebourii (Pojark.) Alexandrova & P.A.Schmidt] Rhododendrontomentosum Harmaja Vacciniummicrocarpum (Turcz.) Schmalh. [\u2261 Oxycoccusmicrocarpus Turcz.] Vacciniummyrtillus L. [\u2261 Vitis-idaeamyrtillus (L.) Moench] Vacciniumuliginosum L. Vacciniumvitis-idaea L. [\u2261 Vitis-idaeavitis-idaea (L.) Britton] Euphorbiaceae51. Juss. (1 genus and 15 species)Euphorbiaalpina C.A.Mey. [7]Euphorbiacaesia Kar. & Kir. [7]Euphorbiaesula L. [= Euphorbiadiscolor Ledeb.] Euphorbiafischeriana Steud. Euphorbiahumifusa Willd. Euphorbiakozlovii Prokh. Euphorbiamacrorhiza Ledeb. Euphorbiamongolica Prokh. Euphorbiapachyrhiza Kar. & Kir. [7]Euphorbiapilosa L. [7]Euphorbiapotaninii Prokh. Euphorbiasoongarica Boiss. [\u2261 Galarhoeussoongaricus (Boiss.) Prokh.] [7]Euphorbiasubcordata C.A.Mey. Euphorbiatshuiensis (Prokh.) Serg. Euphorbiavirgata Waldst. & Kit. [\u2261 Galarhoeusvirgatus Prokh.] Fabaceae52. Lindl. (24 genera and 328 taxa)Alhagimaurorum Medik. Alhagipseudalhagisubsp.kirghisorum (Schrenk) Yakovl. [= Alhagisparsifolia Shap.] SEAmmopiptanthusmongolicus (Maxim.) S.H.Cheng SEAstragalusadmirabilus Pjak & E.Pjak [7]Astragalusadsurgens Pall. [1\u201313]Astragalusagrestis Douglas ex G.Don Astragalusaksaicus Schischk. [7]SEAstragalusalaschanus Bunge [13]Astragalusalberti Bunge Astragalusalbicans Bong. [14]Astragalusalpinus L. Astragalusaltaicola Podlech Astragalusammodytes Pall. Astragalusankylotus Fisch. & C.A.Mey. Astragalusarcuatus Kar. & Kir. [14]Astragalusargutensis Bunge Astragalusarkalycensis Bunge Astragalusaustro-sibiricus Schischk. SEAstragalusbaitagensis Sanchir [14]Astragalusbeketowii (Krassn.) B.Fedtsch. [6]Astragalusborodinii Krasnob. Astragalusbrachybotrys Bunge Astragalusbrevifolius Ledeb. SEAstragalusburtschumensis Sumnev. Astragaluscandidissimus Ledeb. Astragaluschamonobrychis Podlech [7]E Astragaluschangaicus Sanchir E Astragaluschinensis L.f. SEAstragaluschorinensis Bunge [= Astragaluspseudochorinensis N.Ulziykh.] Astragaluschubsugulicus Gontsch. ex N.Ulziykh. [1]E Astragaluscompressus Ledeb. [7]Astragalusconfertus Benth. Astragalusconsanguineus Bong. & C.A.Mey. [10]Astragaluscontortuplicatus L. [14]Astragalusdahuricus Patrin Astragalusdanicus Retz. Astragalusdepauperatus Ledeb. Astragalusdilutus Bunge Astragalusdschimensis Gontsch. Astragalusellipsoideus Ledeb. Astragalusfiliformis (DC.) Poir. [\u2261 Oxytropisfiliformis DC.] Astragalusfollicularis Pall. [3]Astragalusfrigidus A.Gray Astragalusfruticosus Pall. Astragalusgalactites Pall. Astragalusglomeratus Ledeb. Astragalusgobicus Hanelt & Davaz. E Astragalusgranitovii Sanchir E SEAstragalusgregorii B. Fedtsch. & Basil. [7]SEAstragalusgrubovii Sanchir [= Astragalusalaschanensis H.C.Fu] SEAstragalusgrum-grshimailoi Palib. [7]Astragalusgubanovii N.Ulziykh. E SEAstragalushabaheensis Y.X.Liou [14]SEAstragalushamiensis S.B.Ho [= Astragalusbanzragczii N.Ulziykh.] [14]SEAstragalushsinbaticus P.Y.Fu & Y.A.Chen [= Astragalusquasitesticulatus Barratte & Z.Y.Chu] [9]Astragalushypogaeus Ledeb. Astragalusinopinatus Boriss. SEAstragalusjunatovii Sanchir Astragaluskasachstanicus Golosk. [7]Astragaluskaufmannii Krylov Astragaluskenteicus N.Ulziykh. [2]E Astragalusklementzii N.Ulziykh. [3]Astragaluskoslovii B.Fedtsch. & N.Basil. [13]E Astragaluskurtschumensis Bunge Astragaluslaguroides Pall. [= Astragalusgobi-altaicus N.Ulziykh.] Astragaluslasiopetalus Bunge Astragaluslaxmannii Jacq. Astragaluslepsensis Bunge [7]Astragalusleptostachys Pall. [= Astragalusmacropterus DC. = Astragalusmulticaulis Ledeb.] SEAstragaluslupulinus Pall. SEAstragalusluxurians Bunge [7]Astragalusmacrolobus M.Bieb.[= Astragalusmacrocerus C.A.Mey.] Astragalusmacrotrichus E.Peter Astragalusmajevskianus Krylov [7]Astragalusmegalanthus DC. Astragalusmelilotoides Pall. Astragalusminiatus Bunge Astragalusmongholicus Bunge [= Astragalusmembranaceus Fisch. = Astragaluspropinquus Schischk.] Astragalusmonophyllus Bunge [6\u201316]Astragalusnorvegicus Weber SEAstragalusochrias Bunge Astragalusonobrychis L. [10]Astragalusortholobus Bunge [7]Astragalusoxyglottis Steven Astragaluspallasii Spreng. [= Astragaluslasiophyllus Ledeb.] [14]SEAstragaluspavlovii B.Fedtsch. & Basil. [13\u201316]Astragaluspeterae Tsai & Yu Astragalusphysocarpus Ledeb. [7]SEAstragaluspolitovii Krylov [7]SEAstragaluspolozhiae Timokhina SEAstragaluspseudoborodinii S.B.Ho [= Astragalusbaischinticus N.Ulziykh.] [14]Astragaluspseudobrachytropis Gontsch. [6]Astragaluspseudotesticulatus Sanchir [7]E Astragaluspseudovulpinus Sanchir [14]E Astragaluspuberulus Ledeb. [\u2261 Craccinapuberula (Ledeb.) Steven] Astragalusroseus Ledeb. Astragalusrudolffii N.Ulziykh. Astragalusrytidocarpus Ledeb. Astragalussabuletorum Ledeb. Astragalussaichanensis Sanchir E Astragalussanczirii N.Ulziykh. E SEAstragalussaralensis Gontsch. [1]Astragalusscaberrimus Bunge Astragalusscabrisetus Bong. [15]Astragalusschanginianus Pall. [7]Astragalusschrenkianus Fisch. & C.A.Mey. [7]Astragalusscleropodius Ledeb. [7]Astragalussecundus DC. [\u2261 Astragalusfrigidussubsp.secundus (DC.) Vorosch.] Astragalussphaerocystis Bunge [7]Astragalusstenoceras C.A.Mey. [10]Astragalussuffruticosus DC. Astragalussulcatus L. Astragalustamiricus N.Ulziykh. [3]E Astragalustenuis Turcz. [\u2261 AstragalusmelilotoidesPall.var.tenuis (Turcz.) Ledeb.] Astragalustephrolobus Bunge [7]Astragalustibetanus Benth. Astragalustschujensis Bunge [7]Astragalustulinovii B.Fedtsch. [7]SEAstragalustuvinicus Timokhina Astragalusuliginosus L. Astragalusulziykhutagii Sytin [= Astragalusalexandrii N.Ulziykh.] [7]E Astragalusurunguensis N.Ulziykh. [14]SEAstragalusvallestris Kamelin Astragalusvariabilis Bunge Astragalusversicolor Pall. [= Astragalusalexandrii N.Ulziykh. nom. illegit.] Astragalusviridiflavus N.Ulziykh. E Astragalusxanthotrichos Ledeb. [7]SEAstragalusyumenensis S.B.Ho SEAstragaluszacharensis Bunge [9]Astragaluszaissanensis Sumnev. [7]Caraganaarborescens Lam. Caraganabrachypoda Pojark. Caraganabungei Ledeb. SECaraganadavazamcii Sanchir [\u2261 Caraganakorshinskiivar.davazamcii (Sanchir) Yakovlev] Caraganagobica Sanczir E Caraganahalodendron Dum.Cours. [\u2261 Halimodendronhalodendron Voss.] Caraganajubata Poir. SECaraganakorshinskii Kom. Caraganaleucophloea Pojark. Caraganamicrophylla Lam. Caraganapygmaea (L.) DC. [\u2261 Robiniapygmaea L.] [1\u201314]Caraganaspinosa (L.) Vahl Caraganastenophylla Pojark. SECaraganatibetica Kom. Chesneyaferganensis Korsh. [\u2261 Chesniellaferganensis (Korsh.) Boriss.] [15]SEChesneyamongolica Maxim. SEChesniellamacrantha (W.C.Cheng) L.Duan, J.Wen & Zhao Y.Chang [= Spongiocarpellagrubovii (N.Ulziykh.) Yakovlev] Cicersongaricum Steph. [7]Corethrodendronfruticosum B.H.Choi & H.Ohashi [\u2261= Hedysarumfruticosum Pall.] Corethrodendronscoparium (Fisch. & C.A.Mey.) Fisch. & Basiner [\u2261 Hedysarumscoparium Fisch. & C.A.Mey. = Hedysarumarbuscula Maxim.] Glycyrrhizaaspera Pall. Glycyrrhizaglabra L. [= Glycyrrhizaalaschanica Grankina] SEGlycyrrhizainflata Batalin Glycyrrhizapallidiflora Maxim. [9]SEGlycyrrhizasquamulosa Franch. Glycyrrhizauralensis Fisch. [= Glycyrrhizagobica Grankina = Glycyrrhizasoongorica Grankina] Gueldenstaedtiamonophylla Fisch. Gueldenstaedtiaverna (Georgi) Boriss. [= Gueldenstaedtiastenophylla Bunge] Hedysarumalpinum L. [\u2261 Echinolobiumalpinum (L.) Desv.] Hedysarumaustrosibiricum B.Fedtsch. [\u2261 Hedysarumhedysaroidessubsp.austrosibiricum (B.Fedtsch.) Jurtzev] Hedysarumbrachypterum Bunge SEHedysarumchalchorum N.Ulziykh. Hedysarumconsanguineum DC. [7]Hedysarumdahuricum Turcz. [\u2261 Hedysarumgmeliniivar.dahuricum (Turcz.) R.Sha] Hedysarumferganense Korsh. Hedysarumgmelinii Ledeb. Hedysarumhedysaroidessubsp.arcticum (B.Fedtsch.) P.W.Ball [\u2261 Hedysarumarcticum B.Fedtsch.] Hedysarumiliense B.Fedtsch. [7]Hedysaruminundatum Turcz. Hedysarumkamelinii N.Ulziykh. [7]Hedysarumkrylovii Sumn. [7]Hedysarumlintschevskyi Bajtenov Hedysarumneglectum Ledeb. Hedysarumroseum Sims Hedysarumsajanicum N.Ulziykh. [1]SEHedysarumsangilense Krasnob. & Timokhina Hedysarumsetigerum Turcz. Hedysarumtheinum Krasnob. [7]Lathyrushumilis (Ser.) Fisch. [\u2261 Orobushumilis Ser.] Lathyrusledebourii Trautv. [7]LathyruspalustrisL.subsp.pilosus (Cham.) Hult\u00e9n Lathyruspisiformis L. Lathyruspratensis L. Lathyrusquinquenervius (Miq.) Litv. Lespedezabicolor Turcz. [5]Lespedezadaurica (Laxm.) Schindl. [\u2261 Trifoliumdauricum Laxm.] Lespedezajuncea (L.f.) Pers. Lespedezatomentosa Siebold Lotuskrylovii Schischk. & Serg. Medicagofalcata L. Medicagolupulina L. Medicagoplatycarpa (L.) Trautv. Medicagoruthenica Trautv. Melilotusdentatus Pers. Melilotusofficinalis (L.) Lam. Melilotussuaveolens Ledeb. [\u2261 Trigonellasuaveolens (Ledeb.) Coulot & Rabaute] Melilotuswolgicus Poir. Onobrychisarenaria(Kit.)DC.subsparenaria Onobrychisarenariasubsp.sibirica (Turcz.) P.W.Ball [\u2261 Onobrychissibirica (Sirj.) Turcz.] SEOxytropisacanthacea Jurtzev Oxytropisaciphylla Ledeb. SEOxytropisalpestris Schischk. [7]Oxytropisalpicola Turcz. [2]Oxytropisalpina Bunge Oxytropisaltaica Pers. Oxytropisambigua DC. Oxytropisampullata Pers. SEOxytropisbaicalia Pers. SEOxytropisbicolor Bunge [9]Oxytropisbrachycarpa Vassilcz. [7]Oxytropisbungei Kom. E Oxytropiscaerulea DC. Oxytropiscaespitosa Pers. Oxytropiscampanulata Vassilcz. Oxytropischionophylla Schrenk Oxytropisdeflexa DC. Oxytropisdiantha Bunge [= Oxytropischangaica B.Fedtsch. & Basil.] SEOxytropisdubia Turcz. [2]SEOxytropiseriocarpa Bunge Oxytropisfalcata Bunge Oxytropisfragilifolia N.Ulziykh. E SEOxytropisgebleri Fisch. Oxytropisglabra DC. Oxytropisglandulosa Turcz. Oxytropisglareosa Vassilcz. Oxytropisgorbunovii Boriss. Oxytropisgrandiflora DC. Oxytropishailarensis Kitag. SEOxytropisheterophylla Bunge Oxytropishirta Bunge [5]SEOxytropisintermedia Bunge Oxytropisjunatovii Sanchir [13]E SEOxytropisjurtzevii Malyschev [1]Oxytropisklementzii N.Ulziykh. E SEOxytropiskomarovii Vassilcz. SEOxytropiskossinskyi B.Fedtsch. & Basil. Oxytropiskrylovii Schipcz. [7]SEOxytropiskusnetzovii Kryl. & Steinb. Oxytropisladyginii Krylov [7]Oxytropislanata DC. SEOxytropislanuginosa Kom. Oxytropislapponica Gaudin Oxytropislasiopoda Bunge SEOxytropislatibracteata Jurtzev [3]Oxytropislavrenkoi N.Ulziykh. [12]E Oxytropisleptophylla DC. SEOxytropisleucotricha Turcz. Oxytropislongirostra DC. Oxytropismacrosema Bunge SEOxytropismartjanovii Krylov Oxytropismicrantha Bunge E Oxytropismicrophylla DC. SEOxytropismixotriche Bunge SEOxytropismongolica Kom. SEOxytropismonophylla Grubov Oxytropismuricata DC. Oxytropismyriophylla DC. SEOxytropisnitens Turcz. SEOxytropisochrantha Turcz. [9]Oxytropisoligantha Bunge Oxytropisoxyphylla DC. Oxytropispauciflora Bunge Oxytropispavlovii B.Fedtsch. & Basil. E SEOxytropisphysocarpa Ledeb. [7]Oxytropispotaninii Bunge E SEOxytropisprostrata DC. SEOxytropispseudoglandulosa Gontsch. Oxytropispuberula Boriss. Oxytropispumila Fisch. Oxytropisracemosa Turcz. [= Oxytropisgracillima Bunge] Oxytropisrecognita Bunge SEOxytropisreverdattoi Jurtzev SEOxytropisrhizantha Palib. Oxytropisrhynchophysa Schrenk SEOxytropissacciformis H.C.Fu [12]SEOxytropissajanensis Jurtzev Oxytropissaposhnikovii Krylov SEOxytropisselengensis Bunge SEOxytropissetosa DC. Oxytropissongorica DC. [7]Oxytropissordida (Willd.) Pers. [1]Oxytropissquammulosa DC. SEOxytropisstenophylla Bunge Oxytropisstrobilacea Bunge SEOxytropisstukovii Palib. Oxytropissulphurea Ledeb. [7]Oxytropissutaica N.Ulziykh. E Oxytropistenuis Palib. E Oxytropisteres DC. [7]Oxytropistragacanthoides Fisch. Oxytropistrichophysa Bunge SEOxytropistschujae Bunge SEOxytropisturczaninovii Jurtzev Oxytropisulzijchutagii Sanchir [7]E SEOxytropisvarlakovii Serg. Oxytropisviridiflava Kom. E Sophoraalopecuroides L. [12\u201316]Sophoraflavescens Aiton Sphaerophysasalsula DC. Thermopsisalpina Ledeb. SEThermopsisdahurica Czefr. Thermopsislanceolata R.Br. [= Thermopsislanceolatavar.glabra (Czefr.) Yakovlev] Thermopsislongicarpa N.Ulziykh. E Thermopsismongolica Czefr. [\u2261 Thermopsislanceolatavar.mongolica (Czefr.) Q.R.Wang & X.Y.Zhu] SEThermopsisprzewalskii Czefr. Trifoliumeximium Steph. Trifoliumlupinaster L. [1\u20139]Trifoliumpratense L. Trifoliumrepens L. Trigonellaarcuata C.A.Mey. Trigonellacancellata Desf. Viciaamoena Fisch. [= Viciaamoenasubsp.sericea (Kitag.) Kamelin & Gubanov] Viciaamurensis Oett. Viciacostata Ledeb. Viciacracca L. Viciageminiflora Trautv. Viciajaponica A.Gray Viciamacrantha Jurtzev [= Viciamacranthasubsp.olchonensis Peschkova] Viciamegalotropis Ledeb.. Viciamulticaulis Ledeb. [= Vicianervata Sipliv.] SEViciaolchonensis (Peschkova) O.D.Nikif. [\u2261 Viciamacranthasubsp.olchonensis Peschkova] Viciapseudorobus Fisch. & C.A.Mey. Viciaramuliflora (Maxim.) Ohwi [= Viciabaicalensis (Turcz.) B.Fedtsch.] Viciasemenovii B.Fedtsch. Viciatenuifolia Roth SEViciatsydenii Malyshev [4]Viciaunijuga A.Braun [1\u20138]Viciavenosa Maxim. Frankeniaceae53. Desv. (1 genus and 2 species)Frankeniapulverulenta L. [10]SEFrankeniatuvinica Lomon. [10]Gentianaceae54. Juss. (8 genera and 32 taxa)Centauriumpulchellumsubsp.meyeri (Bunge) Tzvelev Centauriumpulchellum(Sw.)Hayeksubsp.pulchellum Comastomafalcatum (Turcz.) Toyokuni Comastomamalyschevii (Zuev) Zuev [\u2261 Gentianellamalyschevii Zuev] Comastomapulmonarium (Turcz.) Toyokuni Comastomatenellum (Rottb.) Toyok. [\u2261 Gentianatenella Rottb.] Gentianaalgida Pall. GentianaaquaticaL.subsp.aquatica Gentianaaquaticavar.pseudoaquatica (Kusn.) S.Agrawal [\u2261 Gentianapseudoaquatica Kusnezow] Gentianadahurica Fisch. [\u2261 Dasystephanadahurica (Fisch.) Zuev] Gentianadecumbens L.f. [\u2261 Dasystephanadecumbens (L.f.) Zuev] Gentianagrandiflora Laxm. Gentianaleucomelaena Maxim. [\u2261 Ciminalisleucomelaena (Maxim.) Zuev] Gentianamacrophylla Pall. Gentianaprostrata Haenke Gentianakarelinii Griseb. [\u2261 Gentianaprostratavar.karelinii (Griseb.) Kusn.] [7]Gentianariparia Kar. & Kir. Gentianasquarrosa Ledeb. [\u2261 Ciminalissquarrosa (Ledeb.) Zuev] [1\u201311]Gentianatriflora Pall. Gentianauniflora Georgi GentianellaamarellaL.subsp.acuta (Michx.) J.M.Gillett [\u2261 Gentianaacuta Michx.] Gentianellaatrata (Bunge) Holub [5]Gentianellaaurea (L.) Harry Sm. Gentianellaturkestanorum (Gand.) Holub Gentianopsisbarbata (Froel.) Ma Haleniacorniculata (L.) Cornaz [\u2261 Swertiacorniculata L.] Lomatogoniumcarinthiacum (Wulfen) Rchb. [\u2261 Swertiacarinthiaca Wulfen] Lomatogoniumrotatum Fr. Swertiabanzragczii Sanchir Swertiadichotoma L. [\u2261 Anagallidiumdichotomum (L.) Griseb.] Swertiamarginata Schrenk [= Swertiakomarovii Pissjauk.] Swertiaobtusa Ledeb. Geraniaceae55. Juss. (2 genera and 19 taxa)Erodiumcicutarium (L.) L\u2019H\u00e9r. Erodiumstephanianum Willd. Erodiumtibetanum Edgew. & Hook.f. Geraniumaffine Ledeb. Geraniumalbiflorum Ledeb. Geraniumamurense Tsyren. Geraniumcollinum Stephan Geraniumdahuricum DC. Geraniumkrylovii Tzvelev Geraniumlaetum Ledeb. Geraniumpamiricum Ikonn. [14]Geraniumplatyanthum Duthie Geraniumpratense L. Geraniumpseudosibiricum J.Mayer Geraniumsaxatile Kar. & Kir. Geraniumsibiricum L. Geraniumtransbaicalicum Serg. Geraniumtransbaicalicumsubsp.turczaninovii (Serg.) Peschkova Geraniumwlassovianum Fisch. Grossulariaceae56. DC. (1 genus and 12 taxa)Ribesaciculare Sm. [\u2261 Grossulariaacicularis (Sm.) Spach] Ribesdiacanthum Pall. [=Ribesdiacanthum f. weichangense J.X.Huang & J.Z.Wang] Ribesfragrans Pall. Ribesgraveolens Bunge Ribesheterotrichum C.A.Mey. Ribesmeyeri Maxim. Ribesnigrum L. Ribespetraeum Wulfen [= Ribesaltissimum Turcz.] Ribesprocumbens Pall. Ribespulchellum Turcz. Ribesrubrum L. Ribesspicatum E.Robson Haloragaceae57. R.Br. (1 genus and 3 species)Myriophyllumsibiricum Kom. [5]Myriophyllumspicatum L. Myriophyllumverticillatum L. Hydrocharitaceae58. Juss. (2 genera and 5 species)Hydrillaverticillata (L.f.) Royle [\u2261 Serpiculaverticillata L.f.] [10]Najasflexilis (Willd.) Rostk. & W.L.E.Schmidt [\u2261 Cauliniaflexilis Willd.] [10]Najasmarina L. Najasminor All. [10]Najastenuissima (A.Braun) Magnus [\u2261 Cauliniatenuissima (A.Braun) Tzvelev] [10]Hypericaceae59. Juss. (1 genus and 4 taxa)HypericumascyronL.subsp.ascyron Hypericumascyronsubsp.gebleri (Ledeb.) N.Robson [\u2261 Hypericumgebleri Ledeb.] [2]Hypericumattenuatum Choisy Hypericumperforatum L. Iridaceae60. Juss. (1 genus and 21 taxa)SEIrisbungei Maxim. [\u2261 Cryptobasisbungei (Maxim.) M.B.Crespo] Irisdichotoma Pall. Irisglaucescens Bunge [6]Irishalophila Pall. [\u2261 Chamaeirishalophila M.B.Crespo] Irishumilis Georgi [= Irisflavissima Pall.] SEIrisivanovae Doronkin SEIriskamelinii Alexeeva Irislactea Pall. [\u2261 Eremirislactea Rodion.] Irisloczyi Kanitz Irisludwigii Maxim. [\u2261 Xyridionludwigii (Maxim.) Rodion.] [7]Irispotaninii Maxim. Irispsammocola Y.T.Zhao [10]SEIrispseudothoroldii Galanin [4]Irisruthenicasubsp.brevituba (Maxim.) Doronkin [\u2261 Irisruthenicavar.brevituba Maxim.] IrisruthenicaKer Gawl.subsp.ruthenica Irisschmakovii Alexeeva [1]E Irissibirica L. [= Irissanguinea Donn] Iristenuifolia Pall. [7\u201315]Iristigridia Bunge Irisuniflora Pall. [\u2261 Jonirisuniflora M.B.Crespo] Irisventricosa Pall. [\u2261 Cryptobasisventricosa M.B.Crespo] Juncaceae61. Juss. (2 genera and 32 taxa)Juncusalpinoarticulatussubsp.fischerianus (V.I.Krecz.) H\u00e4met-Ahti SEJuncusarcticussubsp.grubovii (Novikov) Novikov [\u2261 Juncusgrubovii Novikov] JuncusarticulatusL.subsp.articulatus Juncusarticulatussubsp.limosus (Vorosch.) Vorosch. [\u2261 Juncuslimosus Vorosch.] Juncusbiglumis L. Juncusbufonius L. [1\u201316]Juncuscastaneussubsp.leucochlamys (V.I.Krecz.) Hult\u00e9n [\u2261 Juncusleucochlamys V.I.Krecz.] Juncuscastaneussubsp.triceps (Rostk.) Novikov Juncuscompressus Jacq. Juncusfiliformis L. [7]Juncusgerardi Loisel Juncusgracillimus (Buchenau) V.I.Krecz. & Gontsch. Juncushybridus Brot. [= Juncusbufoniussubsp.ambiguus (Guss.) Schinz & Thell.] Juncusorchonicus Novikov Juncuspersicussubsp.libanoticus (J.Thi\u00e9baut) Novikov & Snogerup [= Juncuslibanoticus J.Thi\u00e9baut] Juncusranarius Songeon & E.P.Perrier [= Juncusbufoniussubsp.nastanthus (V.I.Krecz. & Gontsch.) So\u00f3] Juncussalsuginosus Turcz. Juncussoranthus Schrenk Juncustriglumis L. Juncusturkestanicus V.I.Krecz. & Gontsch. [\u2261 Juncusbufoniussubsp.turkestanicus (V.I.Krecz. & Gontsch.) Novikov] Juncusvirens Buchenau [\u2261 Juncuspapillosusvar.virens (Buchenau) Vorosch.] [4]Luzulaconfusa Lindeb. Luzulamultiflora (Ehrh.) Lej. [3]Luzulamultiflorasubsp.frigida (Buchenau) V.I.Krecz. [7]Luzulamultiflorasubsp.sibirica V.I.Krecz. [\u2261 Luzulasibirica (V.I.Krecz.) V.I.Krecz.] Luzulanivalis (Laest.) Spreng. [\u2261 Luzulacampestrisvar.nivalis Laest.] [1]Luzulapallescens Sw. Luzulaparviflora Desv. Luzulapilosa (L.) Willd. [\u2261 Juncuspilosus L.] [2]Luzularufescensvar.macrocarpa Buchenau [= Luzulachangaica Novikov] [3]Luzulaspicatasubsp.mongolica Novikov LuzularufescensFisch.var.rufescens Juncaginaceae62. Juss. (1 genus and 2 species)Triglochinmaritima L. [1\u201316]Triglochinpalustris L. Lamiaceae63. Martinov (22 genera and 103 taxa)Phlomisoreophila in Mongolia was identified as Phlmoideschinghoensis by Note: The herbarium records of Amethysteacaerulea L. Caryopterismongholica Bunge Dracocephalumargunense Fisch. [5]Dracocephalumdiscolor Bunge Dracocephalumfoetidum Bunge Dracocephalumfragile Turcz. Dracocephalumfruticulosum Stephan Dracocephalumgrandiflorum L. Dracocephalumheterophyllumsubsp.heterophyllum Benth. [3]Dracocephalumheterophyllumsubsp.ovalifolium A.L.Budantzev [\u2261 Dracocephalumovalifolium (A.L.Budantzev) Doronkin] [3]Dracocephalumimberbe Bunge Dracocephalumintegrifolium Bunge [\u2261 Ruyschianaintegrifolia (Bunge) House] Dracocephalumjunatovii A.L.Budantzev Dracocephalummoldavicum C.Morren Dracocephalumnodulosum Rupr. [14]Dracocephalumnutans L. Dracocephalumolchonense Peschkova [4]DracocephalumoriganoidesSteph.subsp.origanoides Dracocephalumoriganoidessubsp.bungeanum Hyl. Elsholtziadensa Benth. Galeopsisbifida Boenn. [\u2261 Galeopsistetrahitvar.bifida (Boenn.) Lej. & Courtois] Hyssopusambiguus (Trautv.) Iljin [= Hyssopusofficinalisvar.ambiguus Trautv.] [7]Hyssopuscuspidatus Boriss. Lagochilusbungei Benth. Lagochilusdiacanthophyllus Benth. Lagochilusilicifolius Bunge Lagopsisdarwiniana Pjak [7]E Lagopsiseriostachya (Benth.) Ikonn.-Gal. Lagopsisflava Kar. & Kir. [7]Lagopsismarrubiastrum (Steph.) Ikonn.-Gal. Lagopsissupina (Steph.) Ikonn.-Gal. Lamiumalbum L. Leonurusdeminutus V.I.Krecz. [\u2261 Leonurusglaucescensvar.deminutus (V.I.Krecz.) Karav.] Leonurusglaucescens Bunge Leonurusmongolicus V.I.Krecz. & Kuprian. Leonuruspseudopanzerioides Krestovsk. [= Leonuruscardiacasubsp.turkestanicus (V.I.Krecz. & Kuprian.) Rech.f.] Leonurussibiricus L. Leonurusturkestanicus V.I.Krecz. & Kuprian. [7]Lophanthuschinensis Benth. Lophanthuskrylovii Lipsky [7]Lycopuslucidus Turcz. [9]Menthaaquatica L. [4]Menthaarvensis L. Menthacanadensis L. [2]Nepetaannua Pall. [\u2261 Schizonepetaannua Schischk.] Nepetadensiflora Kar. & Kir. Nepetamicrantha Bunge Nepetamultifida L. Nepetanuda L. [6]Nepetapungens Benth. [14]Nepetasibirica L. Origanumvulgare L. Panzerinacanescens (Bunge) Soj\u00e1k Panzerinalanata (L.) Soj\u00e1k [\u2261 Ballotalanata Willd.] Phlomoidesagraria (Bunge) Adylov [\u2261 Phlomisagraria Bunge] Phlomoidesalpina Adylov [\u2261 Phlomisalpina Pall.] [7]SEPhlomoideschinghoensis (C.Y.Wu) Kamelin & Makhm. [\u2261 Phlomischinghoensis C.Y.Wu] Phlomoidesmolucelloides (Bunge) Salmaki [\u2261 Eremostachysmolucelloides Bunge] Phlomoidesmongolica (Turcz.) Kamelin & A.L.Budantzev [\u2261 Phlomismongolica Turcz.] Phlomoidespratensis (Kar. & Kir.) Adylov [\u2261 Phlomispratensis Kar. & Kir.] [6]Phlomoidestuberosa Moench [\u2261 Phlomistuberosa L.] [2\u20139]Phlomoidestuvinica (A.Schroet.) Kamelin [\u2261 Phlomistuvinica A.Schroet.] Salviaabrotanoides (Kar.) Sytsma [\u2261 Perovskiaabrotanoides Kar.] [6]Salviadeserta Schangin [6]Scutellariaaltaica Ledeb. [7]Scutellariabaicalensis Georgi Scutellariadependens Maxim. Scutellariagalericulata L. ScutellariagrandifloraSimssubsp.grandiflora Scutellariagrandiflorasubsp.gymnosperma Kamelin & Gubanov E Scutellariakrasevii Kom. & I.Schischk. [3]Scutellariapaulsenii Briq. [7]Scutellariaregelianavar.ikonnikovii (Juz.) C.Y.Wu & H.W.Li Scutellariascordiifolia Fisch. [1\u20139]Scutellariasieversii Bunge Scutellariasupina L. [\u2261 Scutellariaalpinasubsp.supina (L.) I.Richardson] Scutellariatuvensis Juz. [\u2261 Scutellariagrandiflorasubsp.tuvensis (Juz.) Kamelin & Gubanov] [10]Scutellariaviscidula Bunge [9]Stachysasperasubsp.baicalensis (Fisch.) Krestovsk. [\u2261 Stachysbaicalensis Fisch.] Stachyspalustris L. Thymusaltaicus Klokov & Des.-Shost. Thymusbaicalensis Serg. Thymusbituminosus Klokov [1]Thymusdahuricus Serg. Thymusgobi-altaicus (N.Ulziykh.) Kamelin & A.L.Budantzev [13]E Thymusgobicus Tscherneva Thymuskomarovii Serg. [9]Thymusmichaelis Kamelin & A.L.Budantzev Thymusminussinensis Serg. [10]Thymusmongolicus (Ronniger) Ronniger Thymusnarymensis Serg. [7]Thymuspavlovii Serg. Thymusroseus Schipcz. [7]Thymussibiricus Klokov & Des.-Shost. [4]Thymusturczaninovii Serg. [9]ZiziphoraclinopodioidesLam.subsp.clinopodioides Ziziphoraclinopodioidessubsp.bungeana (Juz.) Rech.f. [\u2261 Ziziphorabungeana Juz.] Ziziphorapamiroalaica Juz. Lentibulariaceae64. Rich. (2 genera and 7 species)Pinguiculaalpina L. [1]Pinguiculavulgaris L. [1]Utriculariaaustralis R.Br. Utriculariaintermedia Hayne [\u2261 Lentibulariaintermedia (Hayne) Nieuwl. & Lunell] Utricularia\u00d7japonica Makino [10]Utriculariaminor L. Utriculariavulgaris L. [\u2261 Lentibulariavulgaris (L.) Moench] Liliaceae65. Juss. (5 genera and 15 taxa)Erythroniumsibiricum (Fisch. & C.A.Mey.) Krylov [7]SEFritillariadagana Turcz. Gageabrevistolonifera Levichev [7]Gageafiliformis Merckl. [7]Gageagranulosa Turcz. [7]Gageahiensis Pasch. [= Gageaterraccianoana Pasch.] SEGageakuraiensis Levichev [7]Gageafragifera (Vill.) Ehr.Bayer & G.L\u00f3pez [= Gagealiotardii (Sternb.) Schult. & Schult. f.] [7]Gageapauciflora Turcz. Gageaserotina (L.) Ker Gawl. [\u2261 Lloydiaserotina (L.) Salisb.] Liliumconcolorvar.partheneion (Siebold & de Vriese) Baker [= Liliumbuschianum G.Lodd.] [5]Liliummartagon L. [1\u20137]Liliumpensylvanicum Ker Gawl. [= Liliumdauricum Ker Gawl.] Liliumpumilum Redout\u00e9 [= Liliumpotaninii Vrishcz] Tulipauniflora (L.) Besser Linaceae66. DC. (1 genus and 5 species)Linumaltaicum Ledeb. Linumbaicalense Juz. Linumpallescens Bunge Linumperenne L. Linumviolascens Bunge [7]Lythraceae67. J.St.-Hil. (1 genus and 3 species)Lythrumsalicaria L. [4]Lythrumvirgatum L. Lythrumborysthenicum (Schrank) Litv. [\u2261 Peplisborysthenica Schrank] [10]Malvaceae68. Juss. (2 genera and 5 species)Abutilontheophrasti Medik. [15]Malvaneglecta Wallr.. Malvapusilla Sm. Malvasylvestris L. Malvaverticillata L. Mazaceae69. Reveal (3 genera and 3 species)Mazaceae was separated from Phrymaceae according to Note: Dodartiaorientalis L. Lanceatibetica Hook.f. & Thomson Mazusstachydifolius Maxim. [5]Melanthiaceae70. Batsch (3 genera and 5 species)Anticleasibirica(L.) Kunth [\u2261 Zigadenussibiricus (L.) A.Gray] Parisquadrifolia L. Parisverticillata M.Bieb. Veratrumlobelianum Bernh. Veratrumnigrum L. Menispermaceae71. Juss. (1 genus and 1 species)Menispermumdauricum DC. Menyanthaceae72. Dumort. (2 genera and 2 species)Nymphoidespeltata (S.G.Gmel.) Kuntze Menyanthestrifoliata L. Molluginaceae73. Bartl. (1 genus and 1 species)Hyperteliscerviana (L.) Thulin [\u2261 Mollugocerviana (L.) Ser.] Montiaceae74. Raf. (1 genus and 1 species)Claytoniajoanneana Roem. & Schult. Nitrariaceae75. Lindl. (2 genera and 5 species)Nitrariaroborowskii Kom. [\u2261 Nitrariaschoberivar.roborowskii (Kom.) Grubov] Nitrariasibirica Poir. Nitrariasphaerocarpa Maxim. Peganumharmala L. [= Peganummultisectum (Maxim.) Bobrov] Peganumnigellastrum Bunge Nymphaeaceae76. Salisb. (2 genera and 2 species)Nymphaeatetragona Georgi is not recorded in Mongolia, according to Note: Recently, this family was revised based on field observations and extensive herbarium specimens in Mongolia . NymphaeNupharpumila (Timm) DC. Nymphaeacandida J.Presl. Onagraceae77. Juss. (2 genera and 12 taxa)CircaeaalpinaL.subsp.alpina Circaeaalpinasubsp.caulescens (Kom.) Tatew. [3]Epilobiumanagallidifolium Lam. [= Epilobiumalpinum L.] [7]Epilobiumangustifolium L. [\u2261 Chamaenerionangustifolium (L.) Schur] Epilobiumciliatum Raf. [2]Epilobiumdavuricum Fisch. Epilobiumfastigiato-ramosum Nakai [= Epilobiumbaicalense Popov] Epilobiumhirsutum L. Epilobiumlatifolium L. [= Chamaenerionlatifolium (L.) Sweet] Epilobiumminutiflorum Hausskn. Epilobiumnervosum Boiss. & Buhse [7]Epilobiumpalustre L. Orchidaceae78. Juss. (14 genera and 26 taxa)Note: Orchids of Mongolia were recently revised by Calypso bulbosa (L.) Oakes [= Cypripediumbulbosum L.] Corallorhizatrifida Ch\u00e2tel. Cypripediumcalceolus L.. Cypripediumguttatum Sw. Cypripediummacranthos Sw. Cypripediumxventricosum Sw. [2]Dactylorhizafuchsii (Druce) So\u00f3 Dactylorhizaincarnata (L.) So\u00f3 Dactylorhizaincarnatasubsp.cruenta (O.F.M\u00fcll.) P.D.Sell [3]Dactylorhizasalina (Turcz.) So\u00f3 Dactylorhizaumbrosa (Kar. & Kir.) Nevski Dactylorhizaviridis (L.) R.M.Bateman [= Coeloglossumviride (L.) Hartm.] [1\u20137]Epipogiumaphyllum Sw. Goodyerarepens (L.) R.Br. Gymnadeniaconopsea (L.) R.Br. [\u2261 Orchisconopsea L.] [1\u20135]Herminiumalaschanicum Maxim. [\u2261 Peristylusalaschanicus (Maxim.) N.Pearce & P.J.Cribb] [16]Herminiummonorchis R.Br. Malaxismonophyllos (L.) Sw. [\u2261 Ophrysmonophyllos L.] Neottiacamtschatea Sprengel Neottiapuberula (Maxim.) Szlach. [\u2261 Listerapuberula Maxim.] [5]Orchismilitaris L. Platantherabifolia (L.) Rich. Platantherafuscescens Kraenzl. Platantheraoligantha Turcz. Ponerorchiscucullata (L.) X.H.Jin [\u2261 Neottianthecucullata (L.) Schltr.] Spiranthesaustralis Lindl. Orobanchaceae79. Vent. (9 genera and 57 taxa)Boschniakiarossica (Cham. & Schltdl.) B.Fedtsch. [2]pallida (L.) Spreng. Cistanchedeserticola Ma Cistanchefeddeana K.S.Hao Cistanchelanzhouensis Zhi Y.Zhang [12]Cistanchesalsa (C.A.Mey.) Beck [12\u201316]Cymbariadaurica L. Euphrasiaaltaica Serg. [7]Euphrasiahirtella Jord. Euphrasiamaximowiczii Wettst. Euphrasiapectinata Ten. Euphrasiaschischkinii Serg. [7]Euphrasiasyreitschikovii Govor. Odontitesvulgaris Moench Orobancheamoena C.A.Mey. Orobanchecaesia Rchb. [= Phelypaealanuginosa C.A.Mey. \u2261 Orobanchelanuginosa (C.A.Mey.) Beck] Orobanchecernua Loefl. Orobanchecoerulescens Steph. [\u2261 Orobanchellacoerulescens (Steph.) Piwow.] Orobanchecoerulescensvar.albiflora Kuntze [= Orobanchekorshinskyi Novopokr.] [1\u201315]Orobanchepycnostachya Hance [5]Pedicularisabrotanifolia M.Bieb. Pedicularisachilleifolia Steph. Pedicularisaltaica Steph. Pedicularisamoena Adams Pedicularisanthemifolia Fisch. Pediculariscompacta Steph. Pedicularisdolichorrhiza Schrenk Pediculariselata Willd. SEPedicularisfetisowii Regel [14]Pedicularisfissa Turcz. Pedicularisflava Pall. SEPedicularisincarnata L. [1]Pedicularislabradorica Wirsing Pedicularislapponica L. [1]Pedicularislasiostachys Bunge Pedicularislongiflora Rudolph Pedicularismoschata Maxim. Pedicularismyriophylla Pall. Pedicularisoederi Vahl PedicularispalustrisL.subsp.karoi (Freyn) P.C.Tsoong [\u2261 Pediculariskaroi Freyn] Pedicularisphysocalyx Bunge [7]Pedicularisproboscidea Steven [7]Pedicularisresupinata L. Pedicularisrhinanthoides Schrenk [7]Pedicularisrubens Steph. Pedicularissceptrum-carolinum L. Pedicularissibirica Vved. Pedicularisspicata Pall. Pedicularisstriata Pall. Pedicularissudetica Willd. Pedicularistristis L Pedicularisuliginosa Bunge Pedicularisvenusta Schangin Pedicularisverticillata L. Pediculariswlassoviana Steven [2]Rhinanthusserotinus Oborny [2]Rhinanthussongaricus (Sterneck) B.Fedtsch. [\u2261 Rhinanthusborbasii(D\u00f6rf.)So\u00f3subsp.songaricus (Sterneck) So\u00f3] Oxalidaceae80. R.Br. (1 genus and 1 species)Oxalisacetosella L. Paeoniaceae81. Raf. (1 genus and 3 species)Paeoniaanomala L. Paeoniaintermedia C.A.Mey. Paeonialactiflora Pall. Papaveraceae82. Juss. (6 genera and 30 species)Chelidoniummajus L. Corydalisadunca Maxim. Corydaliscapnoides Pers. Corydalisgrubovii Mikhailova Corydalisimpatiens Fisch. Corydalisinconspicua Bunge Corydalispauciflora Pers. Corydalissajanensis Peschkova [\u2261 Corydalispauciflorasubsp.sajanensis (Peschkova) Mikhailova] [1]Corydalisschanginii B.Fedtsch. Corydalissibirica Pers. Corydalisstricta Steph. [= Corydalisgrubovii Mikhailova] Fumariaofficinalis L. [7]Fumariaschleicheri Soy.-Will. Glauciumelegans Fisch. & C.A.Mey. [14]Glauciumsquamigerum Kar. & Kir. Hypecoumerectum L. Hypecoumlactiflorum (Kar. & Kir.) Pazii Hypecoumleptocarpum Hook.f. & Thomson [3]Papaverbaitagense Kamelin & Gubanov E Papavercanescens Tolm. Papaverchakassicum Peschkova Papaverlapponicum (Tolm.) Nordh. [7]Papavernudicaule L. Papaverpseudocanescens Popov Papaverpseudotenellum Grubov Papaverrefractum (DC.) K.-F.G\u00fcnther [\u2261 Roemeriarefracta DC.] Papaverrubroaurantiacum (Fisch.) C.E.Lundstr. SEPapaversaichanense Grubov [\u2261 Papaverrubroaurantiacumsubsp.saichanense (Grubov) Kamelin & Gubanov] Papaversmirnovii Peschkova [\u2261 Papaverrubroaurantiacumsubsp.smirnovii (Peschkova) Kamelin & Gubanov] Papaversetosum (Tolm.) Peschkova [\u2261 Papaverrubroaurantiacumsubsp.setosum Tolm.] [4]Phyllanthaceae83. Martinov (1 genus and 1 species)Flueggeasuffruticosa Baill. Plantaginaceae84. Juss. (7 genera and 47 species)Callitrichehermaphroditica L. Callitrichepalustris L Hippurisvulgaris L. Lagotisintegrifolia (Willd.) Schischk. Linariaacutiloba Fisch. [\u2261 LinariavulgarisMill.subsp.acutiloba (Fisch.) D.Y.Hong] Linariaaltaica Fisch. Linariaburiatica Turcz. Linariadebilis Kuprian. Linariahepatica Bunge Linariaincompleta Kuprian. [7]Linariamelampyroides Kuprian. Linariapedicellata Kuprian. Plantagoarachnoidea Schrenk [= Plantagolorata (J.Z.Liu) Shipunov \u2261 Plantagoarachnoideavar.lorata J.Z.Liu] [14]Plantagocornuti Gouan Plantagodepressa Willd. Plantagokomarovii Pavlov Plantagomajor L. [2\u201314]Plantagomaritimasubsp.ciliata Printz [= Plantagosalsa Pall.] Plantagominuta Pall. Plantagopolysperma Kar. & Kir. Plantagourvillei Opiz [2]Veronicaanagallis-aquatica L. Veronicaanagalloides Guss. [10]Veronicaarenosa (Serg.) Boriss. [= Veronicalaeta auct. non Kar. & Kir.] Veronicabeccabunga L. Veronicabiloba Schreb. Veronicaciliata Fisch. Veronicadaurica Steven Veronicadensiflora Ledeb. Veronicaferganica Popov Veronicahispidula Boiss. & Huet [= Veronicapusilla Hohen. & Boiss.] [7]Veronicaincana L. Veronicakrylovii Schischkin [9]Veronicalinariifolia Link Veronicalongifolia L. Veronicamacrostemon Bunge Veronicaoxycarpa Boiss. [\u2261 Veronicaanagallis-aquaticasubsp.oxycarpa (Boiss.) A. Jelen] [7]Veronicapinnatasubsp.nana Polozhij [7]VeronicapinnataL.subsp.pinnata Veronicaporphyriana Pavlov Veronicasajanensis Printz [7]Veronicasapozhnikovii Kosachev E Veronicascutellata L. [7]Veronica\u00d7schmakovii Kosachev [7]Veronica\u00d7smirnovii Kosachev & D.A.German [7]Veronicastrumsibiricum (L.) Pennell Veronicastrumtubiflorum (Fisch. & C.A.Mey.) Soj\u00e1k [\u2261 Veronicatubiflora Fisch. & C.A.Mey.] [4]Plumbaginaceae85. Juss. (4 genera and 20 taxa)Armeriamaritimasubsp.sibirica (Turcz.) Nyman [\u2261 Armeriasibirica Turcz.] Goniolimoncallicomum Boiss. Goniolimoneximium Boiss. [7]Goniolimonkrylovii A.V.Grebenjuk Goniolimonspeciosum Boiss. Limoniumaureum (L.) Hill Limoniumbicolor Kuntze Limoniumchrysocomum (Kar. & Kir.) Kuntze Limoniumchrysocomumsubsp.semenovii (Herder) Kamelin Limoniumcongestum Kuntze Limoniumcoralloides (Tausch) Lincz. Limoniumflexuosum Kuntze Limoniumgmelinii Kuntze Limoniumgobicum Ikonn.-Gal. [12]E Limoniumgrubovii Lincz. [9]E Limoniumklementzii Ikonn.-Gal. E Limoniummyrianthum Kuntze [14]Limoniumsuffruticosum Kuntze [14]Limoniumtenellum Kuntze Plumbagellamicrantha (Ledeb.) Spach Poaceae86. Barnhart (58 genera and 229 taxa)Stipa L. was recently revised which included a taxonomic key and species synopsis by Note: The genus Achnatherumcaragana (Trin.) Nevski [= Stipaconferta Poir.] [7]Achnatherumconfusum (Litv.) Tzvelev [\u2261 Stipaconfusa Litv.] SEAchnatheruminebrians (Hance) Keng [\u2261 Stipainebrians Hance] SEAchnatherumpelliotii (Danguy) R\u00f6ser & Hamasha [\u2261 Stipapelliotii Danguy] Achnatherumsibiricum (L.) Keng [\u2261 Stipasibirica (L.) Lam.] [1\u201313]Aeluropuslittoralis (Gouan) Parl. Agropyroncristatum (L.) Gaertn. [\u2261 Bromuscristatus L.] Agropyrondesertorum Schult. Agropyronfragile (Roth) P.Candargy Agropyronkrylovianum Schischk. Agropyronmichnoi Roshev. [\u2261 Agropyroncristatum(L.)Gaertn.subsp.michnoi (Roshev.) \u00c1.L\u00f6ve] Agropyronpumilum (Steud.) P.Candargy Agrostisclavata Trin. [\u2261 Agrostisexaratasubsp.clavata (Trin.) T.Koyama] Agrostisdivaricatissima Mez [= Agrostismongolica Roshev.] Agrostisgigantea Roth Agrostisstolonifera L. SEAgrostistuvinica Peschkova Agrostisvinealis Schreb. [= Agrostistrinii Turcz.] Alopecurusaequalis Sobol. Alopecurusarundinaceus Poir. Alopecurusbrachystachyus M.Bieb. Alopecuruspratensis L. SEAlopecurusturczaninovii O.D.Nikif. Anthoxanthumglabrum (Trin.) Veldkamp [\u2261 Hierochloeglabra Trin.] [1\u201310]Anthoxanthummonticola (Bigelow) Veldkamp [\u2261 Holcusmonticola Bigelow] Anthoxanthumnitens (Weber) Y.Schouten & VeldkampPoanitens Weber = Hierochloeodorata (L.) P.Beauv.] Arctagrostislatifolia Griseb. Arctopoaschischkinii (Tzvelev) Prob. [\u2261 Poaschischkinii Tzvelev] Arctopoatibetica (Munro) Prob. [\u2261 Poatibetica Munro] Aristidaadscensionis L. Arundinellahirta (Thunb.) Tanaka [\u2261 Poahirta Thunb.] Beckmanniasyzigachne Fernald Brachypodiumpinnatum (L.) P.Beauv. [\u2261 Bromuspinnatus L.] Bromusinermis Leyss. Bromusjaponicus Thunb. Bromusoxyodon Schrenk Bromuspumpellianus Scribn. [= Bromuskorotkiji Drobow] Bromusscoparius L. [14]Bromussquarrosus L. [10]Bromustectorum L. Calamagrostisangustifolia Kom. [= Calamagrostisangustifoliasubsp.tenuis (V.N.Vassil.) Tzvelev] Calamagrostisepigejos (L.) Roth [= Calamagrostisepigejossubsp.glomerata (Boiss. & Buhse) Tzvelev] Calamagrostisinexpansa A.Gray [= Calamagrostisinexpansasubsp.micrantha (Kearney) Stebbins] Calamagrostiskorotkyi Litv. [\u2261 Deyeuxiakorotkyi (Litv.) S.M.Phillips & W.L.Chen] SECalamagrostis\u00d7kuznetzovii Tzvelev [4]Calamagrostislapponica (Wahlenb.) Hartm. [\u2261 Arundolapponica Wahlenb.] Calamagrostismacilenta Litv. Calamagrostismacrolepis Litv. [\u2261 Calamagrostisepigejossubsp.macrolepis (Litv.) Tzvelev] Calamagrostisobtusata Trin. Calamagrostispavlovii (Roshev.) Roshev. Calamagrostispseudophragmites Koeler [\u2261 Arundopseudophragmites Haller f.] Calamagrostispurpurea (Trin.) Trin. [\u2261 Arundopurpurea Trin.] SECalamagrostissajanensis Malyschev Calamagrostissalina Tzvelev Calamagrostisstricta (Timm) Koeler [\u2261 Arundostricta Timm] [2]Catabrosaaquatica (L.) P.Beauv. Cenchrusflaccidus (Griseb.) Morrone [\u2261 Pennisetumflaccidum Griseb.] Cinnalatifolia (Trevir.) Griseb. [\u2261 Agrostislatifolia Trevir.] SECleistogenescaespitosa Keng [12]SECleistogenesfestucacea Honda [= Cleistogenesfoliosa Keng] Cleistogeneskitagawae Honda [\u2261 Kengiakitagawae (Honda) Packer] Cleistogenessongorica (Roshev.) Ohwi Cleistogenessquarrosa (Trin.) Keng [2\u201313]Colpodiumaltaicum Trin. DeschampsiacaespitosaP.Beauv.subsp.caespitosa Deschampsiacaespitosasubsp.orientalis Hult\u00e9n Deschampsiacaespitosasubsp.pamirica (Roshev.) Tzvelev [\u2261 Deschampsiapamirica Roshev.] Deschampsiakoelerioides Regel Echinochloacrus-galli (L.) P.Beauv. Elymusbungeanus (Trin.) Melderis Elymusconfusus (Roshev.) Tzvelev [\u2261 Roegneriaconfusa (Roshev.) Nevski] Elymusdahuricus Turcz. Elymusfedtschenkoi Tzvelev [\u2261 Roegneriafedtschenkoi (Tzvelev) J.L.Yang & C.Yen] [7]Elymusgmelinii (Ledeb.) Tzvelev SEElymuskarakabinicus Kotukhov [7]Elymusmacrourus (Turcz.) Tzvelev Elymusmutabilis (Drobow) Tzvelev [= Elymustransbaicalensis (Nevski) Tzvelev = Elymuspraecaespitosus (Nevski) Tzvelev] Elymusnutans Griseb. Elymuspendulinus (Nevski) Tzvelev [= Agropyronvernicosum Nevski = Elymusbrachypodioides (Nevski) Peschkova] Elymusreflexiaristatus (Nevski) Melderis [= Elymusaegilopoides (Drobow) Vorosch.] Elymusrepens (L.) Gould [= Elytrigiarepens (L.) Nevski] Elymusschrenkianus (Fisch. & C.A.Mey.) Tzvelev [= Elymuspamiricus Tzvelev] Elymussibiricus L. Elymusuralensis (Nevski) Tzvelev [= Elymusuralensissubsp.komarovii (Nevski) Tzvelev] Elymusvarius (Keng) Tzvelev [4]Enneapogondesvauxii P.Beauv. Eragrostiscilianensis (All.) Vignolo [10]Eragrostisminor Host Eragrostispilosa (L.) P.Beauv. [= Eragrostispilosasubsp.imberbis (Franch.) Tzvelev] Eremopyrumdistans (K.Koch) Nevski [14]Festucaaltaica Trin. Festucabrachyphylla Schult. & Schult.f. Festucadahurica V.I.Krecz. & Bobr. Festucaextremiorientalis Ohwi [2]SEFestucahubsugulica Krivot. [= Festucasumneviczii Serg.] [1]Festucajacutica Drobow Festucakomarovii Krivot. Festucakryloviana Reverd. Festucakurtschumica E.B.Alexeev [7]Festucalenensis Drobow Festucalitvinovii (Tzvelev) E.B.Alexeev [9]Festucaoreophila Markgr.-Dann. [= Festucavalesiacasubsp.hypsophila (St.-Yves) Tzvelev] Festucaovina L. [= Festucaovinasubsp.sphagnicola (B.Keller) Tzvelev] SEFestucapseudosulcata Drobow [4]Festucarubra L. Festucasibirica Hack. Festucatristis Krylov & Ivanitzk. SEFestucatschujensis Reverd. Festucavalesiaca Schleich. Festucavenusta St.-Yves Glyceriaarundinacea Kunth Glycerialithuanica (Gorski) Gorski Glyceriaspiculosa Roshev. [= Glycerialongiglumis Hand.-Mazz.] Helictochloadahurica (Kom.) Romero Zarco [\u2261 Helictotrichondahuricum (Kom.) Kitag.] Helictochloahookeri (Scribn.) Romero Zarco [= Helictotrichonschellianum (Hack.) Kitag.] Helictotrichondesertorum (Less.) Pilg. [= Helictotrichonaltaicum Tzvelev] Helictotrichonmongolicum (Roshev.) Henrard [\u2261 Avenastrummongolicum (Roshev.) Roshev.] Helictotrichonpubescens (Huds.) Pilg. [= Hordeumbrevisubulatumsubsp.turkestanicum (Nevski) Tzvelev] Hordeumbogdanii Wilensky Hordeumbrevisubulatum Link [= Hordeumbrevisubulatumsubsp.turkestanicum (Nevski) Tzvelev] [1\u201316]Hordeumroshevitzii Bowden [\u2261 Critesionroshevitzii (Bowden) Tzvelev] Koeleriaaltaica (Domin) Krylov Koeleriaasiatica Domin Koeleriaglauca DC. [4]Koelerialitvinowii Domin Koeleriamacrantha (Ledeb.) Schult. Koeleriaspicatasubsp.mongolica (Hult\u00e9n) Barber\u00e1, Quintanar, Soreng, & P.M.Peterson [\u2261 Trisetumspicatumsubsp.mongolicum Hult\u00e9n] SEKoeleriathonii Domin [5]Leymusangustus (Trin.) Pilg. [\u2261 Elymusangustus Trin.] Leymuschinensis (Trin.) Tzvelev SELeymusordensis Peschkova [15]Leymuspaboanus (Claus) Pilg. Leymusracemosus (Lam.) Tzvelev Leymusramosus (K.Richt.) Tzvelev [\u2261 Agropyronramosum K.Richt.] Leymussecalinus (Georgi) Tzvelev [= Leymussecalinusvar.mongolicus (Meld.) Tzvelev = Leymusovatus (Trin.) Tzvelev] Melicanutans L. Melicatranssilvanica Schur [7]Melicaturczaninowiana Ohwi Melicavirgata Turcz. Miliumeffusum L. [2]Nardusstricta L. [3]Neotriniasplendens (Trin.) M.Nobis [\u2261 Achnatherumsplendens (Trin.) Nevski \u2261 Stipasplendens Trin.] Phalarisarundinacea L. Phleumalpinum L. [7]Phleumphleoides (L.) H.Karst. Phragmitesaustralis (Cav.) Steud. [1\u201316]Piptatherumsongaricum (Trin. & Rupr.) Roshev. Poaalpina L. Poaalta Hitchc. [= Poamongolica (Rendle) Keng] Poaaltaica Trin. [\u2261 Poaglaucasubsp.altaica (Trin.) Olonova & G.H.Zhu] Poaangustifolia L. [\u2261 Poapratensissubsp.angustifolia (L.) Dumort.] Poaannua L. [\u2261 Ochlopoaannua (L.) H.Scholz] Poaargunensis Roshev. PoaattenuataTrin.subsp.attenuata . Poaattenuatasubsp.botryoides (Trin.) Tzvelev Poaattenuatasubsp.dahurica (Trin.) Gubanov [\u2261 Poadahurica Trin.] Poaattenuatasubsp.tshuensis (Serg.) Olonova [\u2261 Poaargunensisf.tshuensis Serg.] Poaglauca Vahl Poaircutica Roshev. [4]SEPoakenteica Ivanova Poakrylovii Reverd. [\u2261 Poaurssulensissubsp.krylovii (Reverd.) Olonova] Poanemoralis L. Poapalustris L. Poapratensis L. [= Poapratensissubsp.sabulosa (Turcz.) Tzvelev] Poaraduliformis Prob. Poasibirica Roshevitz Poasmirnowii Roshev. Poasubfastigiata Trin. Poasupina Schrad. Poatianschanica Hack. Poatrivialis L. [1]Poaurssulensis Trin. Poaveresczaginii Tzvelev [7]Poaversicolorsubsp.reverdattoi (Roshev.) Olonova & G.H.Zhu [\u2261 Poareverdattoi Roshev.] PoaversicolorBessersubsp.versicolor [= Poaversicolorsubsp.stepposa (Krylov) Tzvelev] Polypogonmaritimus Willd. Polypogonmonspeliensis (L.) Desf. SEPsammochloavillosa (Trin.) Bor Psathyrostachysjuncea (Fisch.) Nevski Psathyrostachyslanuginosus (Trin.) Nevski Ptilagrostismongholica (Turcz.) Griseb. [\u2261 Stipamongholica Turcz.] Puccinelliaaltaica Tzvelev [14]Puccinelliadistans (Jacq.) Parl. SEPuccinelliafilifolia (Trin.) Tzvelev Puccinelliahackeliana (V.I.Krecz.) V.I.Krecz. [7]Puccinelliahauptiana (V.I.Krecz.) Kitag. Puccinelliamacranthera V.I.Krecz. Puccinelliamanchuriensis Ohwi [3]Puccinellianudiflora (Hack.) Tzvelev SEPuccinelliaprzewalskii Tzvelev [10]Puccinelliaschischkinii Tzvelev Puccinelliatenuiflora Scribn. & Merr. [= Puccinelliakreczetoviczii Bubnova] [1\u201316]Schismusarabicus Nees Schizachnepurpurascenssubsp.callosa (Turcz.) T.Koyama & Kawano Scolochloafestucacea Link [4]Sibirotrisetumsibiricum (Rupr.) Barber\u00e1 [\u2261 Trisetumsibiricum Rupr.] Spodiopogonsibiricus Trin. Sporobolusaculeatus (L.) P.M.Peterson [\u2261 Crypsisaculeata (L.) Aiton] Sporobolusschoenoides (L.) P.M.Peterson [\u2261 Crypsisschoenoides Lam.] Stipaaustromongolica M.Nobis [10]E Stipabaicalensis Roshev. Stipabreviflora Griseb. Stipacapillata L. StipacaucasicaSchmalh.subsp.caucasica Stipacaucasicasubsp.desertorum (Roshev.) Tzvelev [10]Stipaconsanguinea Trin. & Rupr. Stipaglareosaf.pubescens P.A.Smirn. [4\u201312]Stipaglareosa P.A.Smirn. [\u2261 Stipacaucasicasubsp.glareosa (P.A.Smirn.) Tzvelev] Stipagobica Roshev. Stipagrandis P.A.Smirn. Stipakhovdensis L.Q.Zhao E Stipakirghisorum P.A.Smirn. Stipaklemenzii Roshev. Stipakrylovii Roshev. [1\u201314]Stipamongolorum Tzvelev Stipaorientalis Trin. StipapennataL. subsp. pennata Stipapennatasubsp.sabulosa (Pacz.) Tzvelev [10]Stipasareptana A. Beck. Stipasczerbakovii Kotuch. [7]Stipatianschanica Roshev. [= Stipatianschanicasubsp.gobica (Roshev.) D.F. Cui = Stipatianschanicavar.klemenzii (Roshev.) Norl.] Stipazalesskii Wilensky Timouriasaposhnikowii Roshev. [= Stipasaposhnikowii (Roshev.) Kitag.] Tragusmongolorum Ohwi Tripogonchinensis Hack. Tripogonpurpurascens Duthie Trisetumaltaicum Roshev. Zizanialatifolia Turcz. [9]Polemoniaceae87. Juss. (2 genera and 4 species)Phloxsibirica L. Polemoniumboreale Adams Polemoniumchinense Brand Polemoniumpulchellum Bunge Polygalaceae88. Hoffmanns. & Link (1 genus and 3 species)Polygalacomosa Schkuhr [= Polygalahybrida DC.] Polygalasibirica L. Polygalatenuifolia Willd. Polygonaceae89. Juss. (11 genera and 63 taxa)Atraphaxisbracteata Losinsk. Atraphaxiscompacta Ledeb. Atraphaxisfrutescens (L.) K.Koch Atraphaxiskamelinii Yurtseva [14]E Atraphaxispungens Jaub. & Spach [2\u201316]Atraphaxisspinosa L. Atraphaxisvirgata (Regel) Krassn. Bistortaelliptica (Willd.) V.V.Petrovsky SECalligonumebinuricum Ivanova Calligonumjunceum (Fisch. & C.A.Mey.) Litv. Calligonumlitwinowi Drobow [= Calligonumgobicum Losinsk.] Calligonummongolicum Turcz. Fallopiaconvolvulus (L.) \u00c1.L\u00f6ve [\u2261 Polygonumconvolvulus L.] Fallopiadumetorum (L.) Holub [5]Knorringiasibirica(Laxm.)Tzvelevsubsp.sibirica [1\u201316]Knorringiasibiricasubsp.ubsunurica Tzvelev [10]Koenigiaislandica L. Oxyriadigyna Hill Persicariaalpina Gross. Persicariaamphibia (L.) Delarbre Persicariabistorta Samp. [2]Persicariabungeana Nakai Persicariahydropiper (L.) Delarbre [\u2261 Polygonumhydropiper L.] Persicarialapathifolia (L.) Delarbre [\u2261 Polygonumochreatum L.] [1\u201316]Persicaria longiseta var. rotundata (A.J.Li) B.Li [\u2261 Polygonumlongisetumvar.rotundatum A.J.Li] Persicariaminor (Huds.) Opiz Persicariasagittata (L.) H.Gross Persicariavivipara (L.) Ronse Decr. Polygonumabbreviatum Kom. Polygonumalopecuroides Turcz. Polygonumangustifolium Pall. Polygonumarenastrum Boreau Polygonumargyrocoleon Steud. Polygonumaviculare L. Polygonumcognatum Meisn. Polygonumdivaricatum L. Polygonumellipticum Willd. Polygonumhumifusum C.Merck [3]Polygonumintramongolicum Borodina Polygonumnovoascanicum Klokov [14]Polygonumpatulum M.Bieb. Polygonumpolycnemoides Jaub. & Spach Polygonumsericeum Pall. Polygonumtenuissimum A.I.Baranov & Skvortsov [9]Polygonumvalerii A.K.Skvortsov Polygonumvolchovense Tzvelev [7]Rheumcompactum L. Rheumnanum Siev. Rheumrhabarbarum L. [= Rheumundulatum L.] SERheumuninerve Maxim. [13]Rumexacetosa L. Rumexacetosella L. Rumexaquaticus L. Rumexcrispus L. Rumexgmelinii Turcz. Rumexmaritimus L. Rumexmarschallianus Rchb. Rumexpatientia L. Rumexpopovii Pachom. Rumexpseudonatronatus (Borb\u00e1s) Murb. Rumexsimilans Rech.f. Rumexstenophyllus Ledeb. Rumexthyrsiflorus Fingerh. [1\u201314]Potamogetonaceae90. Bercht. & J.Presl (3 genera and 18 taxa)Potamogetonangustifolius Bercht. & J.Presl [10]Potamogetonalpinussubsp.tenuifolius (Raf.) Hulten Potamogetonberchtoldii Fieber Potamogetoncompressus L. Potamogetoncrispus L. Potamogetonfriesii Rupr. Potamogetongramineus L. Potamogetonlucens L. Potamogetonmandschuriensis A.Benn. [5]Potamogetonnatans L. Potamogetonobtusifolius Mert. & W.D.J.Koch Potamogetonperfoliatus L. Potamogetonpraelongus F.Muell. Potamogetonpusillus L. Stuckeniafiliformis (Pers.) B\u00f6rner [\u2261 Potamogetonfiliformis Pers.] Stuckeniapectinata (L.) B\u00f6rner [\u2261 Potamogetonpectinatus L.] Stuckeniavaginata Holub Zannichelliapalustris L. [= Zannichelliapalustrissubsp.pedicellata (Ros\u00e9n & Wahlenb.) Hook.f.] Primulaceae91. Batsch (3 genera and 28 taxa)Primula L. was recently revised by Note: The genus Androsacefedtschenkoi Ovcz. Androsacefiliformis Retz. Androsacegmelinii Gaertn. Androsaceincana Lam. Androsacelactiflora Fisch. [= Androsaceamurensis Prob.] Androsacelehmanniana Spreng. [= Androsacebungeana Schischk. & Bobrov] Androsacelongifolia Turcz. Androsacemaxima L. Androsaceovczinnikovii Schischk. & Bobrov Androsaceseptentrionalis L. AndrosacevillosaL.var.dasyphylla (Bunge) Kar. & Kir. [\u2261 Androsacedasyphylla Bunge] Lysimachiadavurica Ledeb. Lysimachiaeuropaea (L.) U.Manns & Anderb. [\u2261 Trientaliseuropaea L.] Lysimachiamaritima (L.) Galasso [\u2261 Glauxmaritima L.] [1\u201316]Lysimachiathyrsiflora L. [\u2261 Naumburgiathyrsiflora (L.) Rchb.] Primulaalgida Adams Primulabukukunica Kovt. Primulacortusoides L. [3]Primulafarinosa L. Primulalongiscapa Ledeb. Primulamatthiolisubsp.altaica (Losinsk.) Kovt. [\u2261 Cortusaaltaica Losinsk.] Primulamatthiolisubsp.brotheri (Pax) Kovt. [\u2261 Cortusamatthiolif.brotheri Pax] [7]Primulamaximowiczii Regel [5]Primulanivalissubsp.nivalis Pall. Primulanivalissubsp.turkestanica (J.N.Haage & E.Schmidt) Kovt. Primulanivalissubsp.xanthobasis (Fed.) Halda Primulanutans Georgi Primulaserrata Georgi Ranunculaceae92. Juss. (20 genera and 156 taxa)Actea and Cimcifuga, we follow Anemone is considered here according to Pulsatilla at generic level Vorosch.] Aconitumbarbatum Patr. Aconitumbiflorum Fisch. Aconitumcoreanum (H.L\u00e9v.) Rapaics [5]Aconitumdecipiens Vorosch. & Anfalov Aconitumglandulosum Rapaics [=Aconitumaltaicum Steinb. = Aconitumsmirnovii Steinb.] Aconitumgubanovii Luferov & Vorosch. E Aconitumkamelinii A.A.Solovjev [= Aconitumchasmanthum Stapf] E SEAconitumkhanminthunii A.A.Solovjev & Shmakov Aconitumkusnezoffii Rchb. [= Aconitumbirobidshanicum Vorosch.] Aconitumleucostomum Vorosch. Aconitummacrorhynchum Turcz. [4]SEAconitumpaskoi Vorosch. Aconitumranunculoides Turcz. [4]SEAconitumrubicundum (Ser.) Fisch. [\u2261 Aconitumseptentrionalesubsp.rubicundum (Ser.) Vorosch.] [1]Aconitumseptentrionale Koelle SEAconitumturczaninowii Vorosch. Aconitumvolubile Koelle Actaeacimicifuga L. [\u2261 Cimicifugafoetida L.] Actaeadahurica (Turcz.) Franch. [\u2261 Cimicifugadahurica (Turcz.) Maxim.] Actaeaerythrocarpa (Fisch.) Kom. Actaeasimplex Prantl [5]Adonisapennina L. [= Adonissibirica Patrin] Adonismongolica Simonovich E Anemonastrumcrinitum (Juz.) Holub [\u2261 Anemonenarcissiflorasubsp.crinita (Juz.) Kitag.] Anemonastrumdichotomum (L.) Mosyakin [\u2261 Anemonedichotoma L.] Anemonastrumobtusilobum (D.Don) Mosyakin [\u2261 Anemoneobtusiloba Lindl.] [3]Anemonastrumsibiricum (L.) Holub [\u2261 Anemonesibirica L.] Anemonereflexa Steph. Anemonesylvestris L. [\u2261 Anemonoidessylvestris (L.) Galasso] Aquilegiaamurensis Kom. [2]Aquilegiaaradanica Shaulo & Erst [4]Aquilegiadaingolica Erst & Shaulo [7]E Aquilegiaganboldii Kamelin & Gubanov [5]Aquilegiaglandulosa Fisch. Aquilegiagrubovii Erst E Aquilegiajucunda Fisch. & Lallem. [1]Aquilegiasibirica Lam. Aquilegiaviridiflora Pall. Aquilegiaxinjiangensis Erst [7]Callianthemumangustifolium Witasek [7]Callianthemumisopyroides Witasek Callianthemumsajanense Witasek Calthamembranacea (Turcz.) Schipcz. [5]Calthanatans Deyl & Sojak Calthapalustris L. Ceratocephalatesticulata (Crantz) Besser [7]Clematisaethusifolia Turcz. [9]Clematisbrevicaudata DC. SEClematisfruticosa Turcz. Clematisglauca Willd. Clematishexapetala Pall. Clematisintricata Bunge Clematismacropetala Ledeb. [9]Clematisochotensis Poir. & Lam. [4]Clematisorientalis L. [15]Clematissibirica (L.) Mill. Clematissongarica Siev. SEClematistanguticasubsp.mongolica Grey-Wilson [2]ClematistanguticaKorsh.subsp.tangutica Delphiniumaltaicum Nevski Delphiniumbarlykense Lomon. & Khanm. Delphiniumchangaicum N.Friesen E Delphiniumcheilanthum Fisch. Delphiniumcrassifolium Schrad. Delphiniumdictyocarpum DC. [7]SEDelphiniumdissectum Huth Delphiniumelatum L. Delphiniumgrandiflorum L. Delphiniumgubanovii N.Friesen [7]E Delphiniumiliense Huth [14]DelphiniuminconspicuumSerg.subsp.inconspicuum Delphiniuminconspicuumsubsp.mongolicum A.L.Ebel [7]E SEDelphiniummalyschevii N.Friesen [1]Delphiniummirabile Serg. SEDelphiniumsajanense Jurtzev [1]Delphiniumtriste Fisch. Delphiniumukokense Serg. Halerpestessalsuginosa Greene Halerpestessarmentosa (Adams) Kom. & Klob.-Alis Isopyrumanemonoides Kar. & Kir. [7]Leptopyrumfumarioides Rchb. Oxygraphisglacialis (Fisch.) Bunge Paraquilegiaanemonoides Ulbr. Pulsatillaambigua Turcz. SEPulsatillabungeana C.A.Mey. [= Pulsatillabungeanavar.astragalifolia (Pobed.) Grubov] Pulsatillacampanella Fisch. Pulsatilladahurica (Fisch.) Spreng. Pulsatillamultifida (G.Pritz.) Juz. [\u2261 Pulsatillapatenssubsp.multifida (G.Pritz.) Z\u00e4melis] Pulsatillapatens(L.)Mill.subsp.flavescens (Zucc.) Z\u00e4melis [\u2261 Pulsatillaflavescens (Zucc.) Juz.] Pulsatillatenuiloba (Hayek) Juz. Pulsatillaturczaninovii Krylov & Serg. Ranunculusacris L. Ranunculusaltaicus Laxm. Ranunculusaquatilis L. Ranunculusarschantynicus Kamelin, Shmakov & S.V.Smirn. E Ranunculuschinensis Bunge Ranunculuscircinatus Sibth. Ranunculusconfervoides (Fr.) Fr. [= Ranunculustrichophyllussubsp.eradicatus (Laest.) C.D.K.Cook] Ranunculusgmelinii DC. Ranunculusgobicus Maxim. [13]Ranunculusgrandifolius C.A.Mey. [3]Ranunculuskauffmannii Clerc [\u2261 Batrachiumkauffmannii (Clerc) Krecz.] Ranunculuslapponicus L. SERanunculuslasiocarpus C.A.Mey. Ranunculuslingua L. [14]Ranunculuslongicaulis C.A.Mey. Ranunculusmongolicus (Krylov) Serg. [\u2261 Batrachiummongolicum Serg.] Ranunculusmonophyllus Ovcz. [1\u20137]Ranunculusnatans C.A.Mey. Ranunculuspedatifidus Sm. [= Ranunculusrigescens Turcz.] Ranunculuspolyanthemos L. [6]RanunculuspropinquusC.A.Mey.subsp.propinquus Ranunculuspropinquusvar.subborealis (Tzvel.) Luferov Ranunculuspseudohirculus Schrenk SERanunculuspseudomonophyllus Timokhina Ranunculuspulchellus C.A.Mey. Ranunculusradicans C.A.Mey. Ranunculusrepens L. [2\u20139]Ranunculusreptans L. Ranunculussapozhnikovii Schegol. [7]E Ranunculussceleratus L. SERanunculusschmakovii Erst [7]Ranunculussmirnovii Ovcz. [2]Ranunculussulphureussubsp.exaltatus Erst [7]Ranunculustanguticus (Maxim) Ovcz. Ranunculustrautvetterianus Regel [7]Ranunculustrichophyllus Chaix [\u2261 Batrachiumtrichophyllum (Chaix) Bosch = Batrachiumdivaricatum (Schrank) Schur] SERanunculusturczaninovii (Luferov) Vorosch. [2]SERanunculustuvinicus Erst [7]Thalictrumalpinum L. Thalictrumbaicalense Turcz. Thalictrumcontortum L. [= Thalictrumaquilegiifoliumvar.sibiricum Regel & Tiling] Thalictrumfoetidum L. Thalictrumisopyroides C.A.Mey. SEThalictrumminussubsp.appendiculatum (C.A.Mey.) Gubanov [\u2261 Thalictrumappendiculatum C.A.Mey.] [3]Thalictrumminussubsp.elatum (Jacq.) Stoj. & Stef. [= Thalictrumminussubsp.kemense (Fries) Cajander] [3]ThalictrumminusL.subsp.minus Thalictrumpetaloideum L. SEThalictrumschischkinii N.Friesen [= Thalictrumaltaicum (Schischk.) Serg.] [7]Thalictrumsimplex L. Thalictrumsquarrosum Steph. Trolliusaltaicus C.A.Mey. Trolliusasiaticus L. Trolliusaustrosibiricus Erst & Luferov [7]Trolliuschinensis Bunge [7]Trolliusdschungaricus Regel [14]Trolliusledebourii Rchb. Trolliuslilacinus Bunge Trolliussajanensis Sipliv. [1]Trolliussibiricus Schipcz. [5]Trolliusvicarius Sipliv. [5]Rhamnaceae93. Juss. (1 genus and 5 species)Rhamnusdavurica Pall. Rhamnuserythroxylon Pall. Rhamnusmaximovicziana J.J.Vassil. Rhamnusparvifolia Bunge Rhamnusutilis Decne. Rosaceae94. Juss. (28 genera and 168 taxa)Agrimoniapilosa Ledeb. Alchemillaargutiserrata H.Lindb. [7]Alchemillachangaica V.N.Tikhom. E Alchemillacircularis Juz. [7]Alchemillacyrtopleura Juz. Alchemillaflavescens Buser [3]Alchemillagracilis Pax [3]Alchemillagubanovii V.N.Tikhom. Alchemillahebescens Juz. Alchemillakrylovii Juz. [7]Alchemillamurbeckiana Buser [7]SEAlchemillapavlovii Juz. anserina (L.) Rydb. [\u2261 Potentillaanserina L.] Aruncussylvester Kostel. [5]Chamaerhodosaltaica Bunge SEChamaerhodoscorymbosa Murav. Chamaerhodoserecta (L.) Bunge [1\u201313]Chamaerhodosgrandiflora Ledeb. [5]Chamaerhodossabulosa Bunge Chamaerhodostrifida Ledeb. Coluriageoides Ledeb. [\u2261 Dryasgeoides Pall.] Comarumpalustre L. Cotoneastermegalocarpus Popov [7]Cotoneastermelanocarpus Lodd. Cotoneastermongolicus Pojark. Cotoneasterneopopovii Czerep. [4]Cotoneasteruniflorus Bunge Crataegusdahurica Koehne Crataegusmaximowiczii C.K.Schneid. [5]Crataegussanguinea Pall. Dasiphorafruticosa (L.) Rydb. [\u2261 Potentillafruticosa L.] Dasiphoraparvifolia (Fisch.) Juz. [\u2261 Potentillaparvifolia Fisch.] Dryasgrandis Juz. Dryasincisa Juz. [1]Dryasoxyodonta Juz. Dryaspunctata Juz. SEDryassumneviczii Serg. [1]Farinopsissalesoviana (Steph.) Chrtek & Soj\u00e1k [\u2261 Comarumsalesovianum (Steph.) Ledeb] Filipendulaangustiloba Maxim. Filipendulapalmata Maxim. Filipendulaulmaria (L.) Maxim. Fragariaorientalis Losinsk. Fragariaviridis Weston [2]Geumaleppicum Jacq. Geumrivale L. [7]Malusbaccata (L.) Borkh. SEPotaniniamongolica Maxim. Potentillaacaulis L. Potentillaacervata Soj\u00e1k [= Potentillachenteica Soj\u00e1k] Potentillaagrimonioides M.Bieb. [= Potentillalydiae Kurbatski] Potentillaaltaica Bunge [= PotentillaniveaL.var.pinnatifida Lehm.] [7]Potentillaangustiloba T.T.Yu & C.L.Li Potentillaaphanes Soj\u00e1k Potentillaarenosa (Turcz.) Juz. [\u2261 Potentillaniveavar.arenosa Turcz.] Potentillaasiatica (Th.Wolf) Juz. [7]Potentillaastragalifolia Bunge SEPotentilla\u00d7burjatica Soj\u00e1k [2]Potentillachalchorum Soj\u00e1k SEPotentilla\u00d7chamaeleo Soj\u00e1k Potentillachinensis Ser. Potentillachionea Soj\u00e1k Potentillachrysantha Trevir. Potentillaconferta Bunge Potentillacoriacea Soj\u00e1k [3]E Potentillacrantzii (Crantz) Fritsch [7]Potentillacrebridens Juz. [= Potentillaniveavar.elongata Th.Wolf] Potentilladesertorum Bunge SEPotentilla\u00d7drymeja Soj\u00e1k Potentillaekaterinae Kamelin ex Kechaykin [13]E Potentillaelegans Cham. & Schltdl. [1]Potentillaelegantissima Polozhij [3]Potentillaevestita Th.Wolf Potentillaexuta Soj\u00e1k Potentillaflagellaris D.F.K.Schltdl. Potentillafragarioides L. Potentillagelida C.A.Mey. Potentillagobica Soj\u00e1k [14]E SEPotentillagracillima Kamelin Potentillahilbigii Soj\u00e1k [3]E Potentillahubsugulica Soj\u00e1k [1]E Potentillaikonnikovii Juz. E Potentillainopinata Soj\u00e1k E Potentillajenissejensis Polozhij & W.Smirnova [= PotentillaagrimonioidesM.Bieb.var.kobdoensis Soj\u00e1k] Potentillakryloviana Th.Wolf Potentillalaevipes Soj\u00e1k [7]E Potentillalaevissima Kamelin [7]E Potentillaleucophylla Pall. [= Potentillabetonicifolia Poir.] Potentillalongifolia D.F.K.Schltdl. [1\u201313]Potentillamongolica Krasch. E Potentillamulticaulis Bunge Potentillamultifida L. [= Potentillatenella Turcz.] [1\u201314]Potentillanivea L. Potentillanorvegica L. [= Potentillamonspeliensis L.] Potentillanudicaulis D.F.K.Schltdl. [= Potentillastrigosa Pall.] [12]SEPotentilla\u00d7olchonensis Peschkova [6]Potentillaornithopoda Tausch SEPotentillaozjorensis Peschkova Potentillapamirica Th.Wolf Potentillapamiroalaica Juz. [14]Potentillapensylvanica L. [\u2261 Pentaphyllumpennsylvanicum (L.) Lunell] Potentillaregeliana Th.Wolf [6]SEPotentilla\u00d7rhipidophylla Soj\u00e1k [3]SEPotentillarigidula Th.Wolf Potentillasanguisorba D.F.K.Schltdl. Potentillaschmakovii Kechaykin E SEPotentillasergievskajae Peschkova Potentillasericea L. SEPotentillaserrata Soj\u00e1k [3]SEPotentillasischanensis Bunge Potentillasongorica Bunge SEPotentillastepposa Soj\u00e1k Potentillasubdigitata T.T.Yu & C.L.Li [= Potentillajunatovii Rudaya & A.L.Ebel] [7]Potentillasupina L. Potentillatanacetifolia D.F.K.Schltdl. Potentillatergemina Soj\u00e1k SEPotentillatericholica Sobolevsk. Potentillatetrandra (Bunge) Hook.f. [\u2261 Sibbaldiatetrandra Bunge] Potentillaturczaninowiana Stschegl. Potentillaturkestanica Soj\u00e1k Potentillatytthantha (Soj\u00e1k) Kechaykin E Potentilla\u00d7vanzhilii Gundegmaa & Kechaykin [3]E Potentillaverticillaris Stephan Potentillavirgata Lehm. Prunusmongolica Maxim. [\u2261 Amygdalusmongolica (Maxim.) Ricker] Prunuspadus L. Prunuspedunculata Maxim. [\u2261 Amygdaluspedunculata Pall.] Prunussibirica L. [\u2261 Armeniacasibirica (L.) Lam.] Rosaacicularis Lindl. Rosaalbertii Regel [7]Rosabaitagensis Kamelin & Gubanov [14]E Rosabeggeriana Schrenk [14]Rosadavurica Pall. Rosakokanica (Regel) Regel [7]Rosalaxavar.kaschgarica (Rupr.) Y.L.Han [= Rosakaschgarica Rupr.] RosalaxaLindl.var.laxa Rosaoxyacantha M.Bieb. Rosaplatyacantha Schrenk [14]Rosaspinosissima L. Rosaxanthina Lindl. [9]Rubusarcticus L. Rubuschamaemorus L. [2]Rubushumilifolius C.A.Mey. Rubussachalinensis H.L\u00e9v. Rubussaxatilis L. Sanguisorbaalpina Bunge Sanguisorbaofficinalis L. [1\u201311]Sanguisorbaparviflora (Maxim.) Takeda [9]Sanguisorbatenuifolia Fisch. Sibbaldiaprocumbens L. Sibbaldiantheadpressa (Bunge) Juz. [\u2261 Sibbaldiaadpressa Bunge] Sibbaldianthebifurca (L.) Kurtto & T.Erikss. [\u2261 Potentillabifurca L.] [1\u201314]Sibbaldiantheimbricata (Kar. & Kir.) Mosyakin & Shiyan [\u2261 Potentillaimbricata Kar. & Kir.] Sibbaldiantheorientalis (Soj\u00e1k) Mosyakin & Shiyan [= Potentillabifurcavar.major Ledeb.] Sibbaldianthesemiglabra (Soj\u00e1k) Mosyakin & Shiyan [\u2261 Potentillasemiglabra Juz.] SESibbaldianthesericea Grubov Sibiraealaevigata (L.) Maxim. [7]Sorbariasorbifolia (L.) A.Braun SorbusaucupariaL.subsp.glabrata (Wimm. & Grab.) Hedl. [= Sorbussibirica Hedl.] Spiraeaalpina Pall. Spiraeaaquilegiifolia Pall. Spiraeachamaedryfolia L. [5]Spiraeadahurica (Rupr.) Maxim. Spiraeaelegans Pojark. [4]Spiraeaflexuosa Fisch. Spiraeahypericifolia L. SpiraeamediaF.Schmidtsubsp.media [= Spiraeasericea Turcz.] Spiraeapubescens Turcz. Spiraeasalicifolia L. Rubiaceae95. Juss. (3 genera and 13 taxa)Asperulagobicola Grubov [= Asperulasaxicola Grubov] E Galiumamblyophyllum Schrenk Galiumboreale L. Galiumdahuricum Turcz. [2]Galiumdensiflorum Ledeb. Galiumhumifusum M.Bieb. Galiumsongaricum Schrenk Galiumspurium L. Galiumtrifidum L. Galiumuliginosum L. GaliumverumL.subsp.verum [= Galiumdensiflorum Ledeb.] Galiumverumsubsp.wirtgenii (F.W.Schultz) Oborny [7]Rubiacordifolia L. [\u2261 Galiumcordifolium (L.) Kuntze] Ruppiaceae96. Horan. (1 genus and 1 species)Ruppiamaritima L. [10]Rutaceae97. Juss. (2 genera and 2 species)Haplophyllumdauricum (L.) G.Don Dictamnusalbus L. Salicaceae98. Mirb. (2 genera and 47 species)Populuseuphratica Olivier [12\u201316]Populuslaurifolia Ledeb. [= Populuspilosa Rehder] Populussimonii Carri\u00e8re [9]Populussuaveolens Fisch. Populustremula L. Salixabscondita Laksch. Salixalatavica Kar. Salixarctica Pall. Salixbebbiana Sarg. Salixberberifolia Pall. Salixbrachypoda (Trautv. & C.A.Mey.) Kom. Salixcaesia Vill. Salixdivaricata Pall. Salixglauca L. Salixgmelinii Pall. [= Salixdasyclados Wimmer] Salixgordejevii Y.L.Chang & Skvortsov Salixhastata L. Salixjenisseensis (F.Schmidt) Flod. Salixkochiana Trautv. Salixledebouriana Trautv. Salixmicrostachya Turcz. Salixmiyabeana Seemen Salixmyrtilloides L. SESalixnasarovii A.K.Skvortsov [1]Salixnipponica Franch. & Sav. [9]Salixnummularia Andersson Salixpolaris Wahlenb. [1]Salixpseudopentandra (Flod.) Flod. [= Salixpentandravar.intermedia Nakai] Salixpyrolifolia Ledeb. Salixrectijulis Ledeb. Salixrecurvigemmata A.K.Skvortsov [= Salixrecurvigemmis A.K.Skvortsov] Salixreticulata L. Salixrhamnifolia Pall. Salixrorida Laksch. Salixrosmarinifolia L. Salixsajanensis Nasarow Salixsaposhnikovii A.K.Skvortsov Salixsaxatilis Turcz. Salixschwerinii E.L.Wolf Salixtaraikensis Kimura Salixtenuijulis Ledeb. Salixtriandra L. Salixturanica Nasarow Salixturczaninowii Laksch. Salixudensis Trautv. & C.A.Mey. [9]Salixvestita Pursh Salixviminalis L. Santalaceae99. R.Br. (1 genus and 6 species)Thesiumchinense Turcz. [9]Thesiumlongifolium Turcz. Thesiumrefractum C.A.Mey. Thesiumrepens Ledeb. SEThesiumsaxatile Turcz. SEThesiumtuvense Krasnob. Saxifragaceae100. Juss. (5 genera and 21 taxa)Bergeniacrassifolia (L.) Fritsch Chrysospleniumnudicaule Bunge [6]SEChrysospleniumpeltatum Turcz. SEChrysospleniumsedakowii Turcz. Chrysospleniumserreanum Hand.-Mazz. [= Chrysospleniumalternifoliumsubsp.sibiricum (Ser.) Hult\u00e9n] Micranthesdavurica (Willd.) Small [\u2261 Saxifragadavurica Willd.] [2]Micranthesfoliolosa (R.Br.) Gornall [\u2261 Saxifragafoliolosa R.Br.] Micrantheshieraciifolia Haw. [\u2261 Saxifragahieraciifolia Waldst. & Kit.] Micranthesmelaleuca (Fisch.) Losinsk. [\u2261 Saxifragamelaleuca Fisch.] Micranthesnelsonianasubsp.aestivalis (Fisch. & C.A.Mey.) Elven & D.F.Murray [\u2261 Saxifragaaestivalis Fisch. & C.A.Mey.] Micranthesnivalis (L.) Small [\u2261 Saxifraganivalis L.] [1]Mitellanuda L. Saxifragabronchialis L. Saxifragacernua L. Saxifragahirculus L. Saxifragamacrocalyx Tolm. [= Saxifragaflagellaris Willd.] SaxifragaoppositifoliaL.subsp.oppositifolia [= Saxifragaasiatica Hayek] Saxifragasetigera Pursh Saxifragasibirica L. Saxifragaterektensis Bunge Scheuchzeriaceae101. F.Rudolphi (1 genus and 1 species)Scheuchzeriapalustris L. [2]Scrophulariaceae102. Juss. (3 genera and 6 species)Limosellaaquatica L. Scrophulariaaltaica Murray Scrophulariacanescens Bong. [= Scrophulariahilbigii J\u00e4ger] Scrophulariaincisa Weinm. Scrophulariaumbrosa Dumort. [10]Verbascumthapsus L. [4]Solanaceae103. Juss. (4 genera and 9 taxa)Hyoscyamusniger L. Hyoscyamuspusillus L. Lyciumchinensevar.potaninii (Pojark.) A.M.Lu [\u2261 Lyciumpotaninii Pojark.] [16]Lyciumruthenicum Murray Lyciumtruncatum Y.C.Wang Physochlainaalbiflora Grubov E Physochlainaphysaloides (L.) G.Don Solanumkitagawae Sch\u00f6nb.-Tem. Solanumseptemlobum Bunge Tamaricaceae104. Link (3 genera and 13 taxa)Myricariabracteata Royle Myricarialongifolia Ehrenb. Reaumuriasoongarica Maxim. Tamarixarceuthoides Bunge Tamarixelongata Ledeb. Tamarixgracilis Willd. Tamarixhispida Willd. [13]Tamarix\u00d7karelinii Bunge Tamarixkasahorum Gorschk. Tamarixlaxa Willd. Tamarixleptostachya Bunge Tamarixramosissima Ledeb. Tamarixsmyrnensis Bunge Thymelaeaceae105. Juss. (2 genera and 3 species)Diarthronaltaicum (Thi\u00e9b.-Bern.) Kit Tan [\u2261 Stelleraaltaica Thi\u00e9b.-Bern.] [7]Diarthronlinifolium Turcz. Stellerachamaejasme L. Tofieldiaceae106. Takht. (1 genus and 1 species)Tofieldiacoccinea Richardson [1]Typhaceae107. Juss. (2 genera and 12 species)Sparganiumemersum Rehmann Sparganiumglomeratum (Laest.) Beurl. Sparganiumnatans L. Sparganiumstoloniferum (Graebn.) Buch.-Ham. Typhaangustifolia L. Typhadomingensis Pers. Typhajoannis Mavrodiev [9]Typhalatifolia L. Typhalaxmannii Lepech. Typhaminima Funck Typhaorientalis C.Presl [5]Typhatzvelevii Mavrodiev [4]Ulmaceae108. Mirb. (1 genus and 3 taxa)Ulmusdavidianavar.japonica (Rehder) Nakai Ulmusmacrocarpa Hance Ulmuspumila L. Urticaceae109. Juss. (2 genera and 4 taxa)Parietariadebilis G.Forst. Urticaangustifolia Fisch. Urticacannabina L. UrticadioicaL.subsp.sondenii (Simm.) Hyl. [\u2261 Urticasondenii (Simm.) Avror] Violaceae110. Batsch. (1 genus and 27 taxa)Viola L in the Russian Far East and adjacent territories. In this study, accepted species and nomenclature mostly follow Note: Recently, Violaacuminata Ledeb. SEViolaalexandrowiana (W.Becker) Juz. [4]Violaaltaica Ker Gawl. Violaarvensis Murray [4]Violabiflora L. Violabrachyceras Turcz. [2]Violacollina Besser Violadactyloides Schult. Violadisiuncta W.Becker [7]Violadissecta Ledeb. Violaepipsiloides \u00c1.L\u00f6ve & D.L\u00f6ve Violagmeliniana Schult. Violaincisa Turcz. SEViolaircutiana Turcz. [2]Violamacroceras Bunge [7]Violamauritii Teplouchow Violamirabilis L. Violanemoralis Kuetz. [2]Violapatrinii Ging. Violarudolfii Vl.V.Nikitin Violarupestris F.W.Schmidt Violasacchalinensis H.Boissieu Viola\u00d7schauloi Vl.V.Nikitin Violaselkirkii Pursh [2]Violatenuicornissubsp.trichosepala W.Becker [4]Violauniflora L. Violavariegata Fisch. Zygophyllaceae111. R.Br. (2 genera and 13 taxa)Tribulusterrestris L. Zygophyllumbrachypterum Kar. & Kir. Zygophyllumgobicum Maxim. Zygophyllumkaschgaricum Boriss. [\u2261 Sarcozygiumkaschgaricum (Boriss.) Y.X.Liou] [12\u201316]Zygophyllummacropterum C.A.Mey. [= Zygophyllumpinnatum Cham. & Schltdl.] SEZygophyllummelongena Bunge SEZygophyllummucronatum Maxim. Zygophyllumneglectum Grubov E Zygophyllumpotaninii Maxim. Zygophyllumpterocarpum Bunge Zygophyllumrosowiivar.latifolium (Schrenk) Popov [13\u201316]ZygophyllumrosowiiBungevar.rosowii Zygophyllumxanthoxylon (Bunge) Maxim."} +{"text": "Streptococcus episodes was misstated in Invasive Group B Streptococcus Infections in Adults, England, 2015\u20132016 . The correct rate is 4.09/10,000 live births. The article has been corrected online (https://wwwnc.cdc.gov/eid/article/26/6/19-1141_article).The rate of pregnancy-related invasive group B"} +{"text": "In the originally published version of the below listed manuscripts, acknowledgement of NIH/NIMH funding was omitted in error. These errors have been corrected.https://doi.org/10.1093/psyrad/kkab009https://doi.org/10.1093/psyrad/kkab017"} +{"text": "Some of these LDs establish close contact sites with the outer mitochondrial membrane (OMM) (S. cerevisiae . C,5. CS. crevisiae . Recent revisiae . Ectopic stimuli , thereby stimuli . Therefo"} +{"text": "This article has been corrected: In 14359-14373. https://doi.org/10.18632/oncotarget.8736Original article: Oncotarget. 2017; 8:14359\u201314373."} +{"text": "Redistributing deaths by ill-defined and unspecified causes on cancer mortality in Brazil\u201d, DOI https://doi.org/10.11606/s1518-8787.2021055003319, published on the Revista de Sa\u00fade P\u00fablica. 2021;55:106, on page 1, where it reads:In the article \u201cDESCRIPTORS: Chemical Waste. Hazardous Waste. Product Labeling. Substances, Products and Materials Transportation.It should read as follows:DESCRIPTORS: Neoplasms, mortality. Data Accuracy. Vital Statistics. Cause of Death."} +{"text": "This article has been corrected: In 115803-115816. https://doi.org/10.18632/oncotarget.23253Original article: Oncotarget. 2017; 8:115803\u2013115816."} +{"text": "This article has been corrected: In 1143-1156. https://doi.org/10.18632/oncotarget.2732Original article: Oncotarget. 2015; 6:1143\u20131156."} +{"text": "The first and third authors\u2019 names are spelled incorrectly. The correct names are Md. Alamgir Sarder and Md. Maniruzzaman.https://doi.org/10.1371/journal.pone.0245923Sarder MA, Islam SMS, Maniruzzaman M, Talukder A, Ahammed B (2021) Prevalence of unintended pregnancy and its associated factors: Evidence from six south Asian countries. PLoS ONE 16(2): e0245923."} +{"text": "Correction to: BMC Health Serv Res 21, 474 (2021)https://doi.org/10.1186/s12913-021-06459-4Following publication of the original article , the autThe incorrect author name is: L. E. U. N. G. Ling YanThe correct author name is: Ling Yan LEUNGThe author group has been updated above and the original article has been"} +{"text": "J Clin Invest. 2012;122(11):3955\u20133959. https://doi.org/10.1172/JCI63113Original citation: J Clin Invest. 2022;132(1):e157161. https://doi.org/10.1172/JCI157161Citation for this corrigendum: GrnKO was incorrect. The correct sequence is below.The sense primer 1 listed for genotyping 5\u2032-AGTGGGGCTGGCCATCCTCThe authors regret the error."} +{"text": "This article has been corrected: In 28523-28539. https://doi.org/10.18632/oncotarget.8660Original article: Oncotarget. 2016; 7:28523\u201328539."} +{"text": "It is with a great pleasure to announce the top three most-cited AVD articles on Web of Science.Review ArticleVisceral Artery Aneurysms and Pseudoaneurysms? Should They All be Managed by Endovascular Techniques?Alfredo C. Cordova and Bauer E. SumpioCitation: 59Published: 2013 (Vol. 6 No. 4: 687-693)DOI: 10.3400/avd.ra.13-00045Review ArticleCompression Therapy: Clinical and Experimental EvidenceHugo PartschCitation: 56Published: 2012 (Vol. 5 No. 4: 416-422)DOI: 10.3400/avd.ra.12.00068Review ArticleThe Relationship between Vascular Function and the Autonomic Nervous SystemEisuke Amiya, Masafumi Watanabe, and Issei KomuroCitation: 46Published: 2014 (Vol. 7 No. 2: 109-119)DOI: 10.3400/avd.ra.14-00048"} +{"text": "This article has been corrected: In 32917-32928. https://doi.org/10.18632/oncotarget.25952Original article: Oncotarget. 2018; 9:32917\u201332928."} +{"text": "JCI Insight. 2021;6(9):e144260. https://doi.org/10.1172/jci.insight.144260Original citation: JCI Insight. 2022;7(6):e159640. https://doi.org/10.1172/jci.insight.e159640Citation for this corrigendum: Information on data availability was omitted from Methods. The correct information is below.Data availabilityRaw data were deposited in the National Genomics Data Center\u2019s Genome Sequence Archive .The authors regret the error."} +{"text": "AbdiLisa Abernathy-CloseHamada A. AboubakrJ\u00f4natas S. Abrah\u00e3oZaky AdamBeth M. AdamowiczAmeeta K. AgarwalSurya D. AggarwalDaniel Aguirre de CarcerMir Alvee AhmedCaio Augusto Martins AiresSamuel L. AitkenAshok A. AiyarKola AjuwonRobert A. AkinsMd. Tauqeer AlamBegum AlaybeyogluLuis D. AlcarazGajender AletiGladys AlexandreNahid AliShaukat AliDaniela S. Aliaga GoltsmanJoannie M. AllaireJonathan AllenRey Custer AllenAlexandre AlmeidaHassan M. Al-TameemiEva AlvarezStefano AmalfitanoAman Aman KhanKatherine R. AmatoAlongkorn AmnuaykanjanasinAmitesh AnandCheryl P. AndamAnnette Carola AndersonMichael AndersonRika AndersonSimon C. AndrewsS. Andreas AngermayrNana Y. D. AnkrahAlessio AprileElizabeth ArchieFarnoosh ArfaeeH\u00e9ctor ArguelloJos\u00e9 M. Arg\u00fcelloGunjan AroraMelinda Marie AshcroftAlexander AskenovContreras AsuncionHaruyuki AtomiJennifer M. AuchtungSelcan AydinFrank O. AylwardTaj AzarianReha O. AzizogluPaul BabitzkeGiovanni BacciVarsha Dave BadalBrian D. BadgleyJin-Woo BaeJake BaileyBrett J. BakerRoberto Balbont\u00ednRegina Lucia BaldiniDavid A. BaltrusMarcy J. BalunasHoria Leonard BanciuSreejata BandopadhyayNoam BarDaniel BarkanFrancisco Barona-G\u00f3mezRodolphe BarrangouDouglas H. BartlettNick BartsMarcelo Bueno BatistaSilvia Beatriz BatistaDavid L. V. BauerAndreas J. B\u00e4umlerJoseph BaurMatt BawnGwyn A. BeattieWilliam N. BeaversTiago BeitesGeorgios N. BelibasakisAeriel D. BelkAmanda BendiaJose A. BengoecheaMaureen BergTeresa M. BergholzKathryn BernardAude BernheimAnthony BertagnolliMatthew BertinElizabeth N. BessSharon BewickOliver K. I. BezuidtMohini BhattacharyaGiancarlo A. BiaginiLewis BingleJordan E. BisanzPradeep BistPinaki BiswasEran BlacherJonathan M. BlackburnChristopher BlackwoodJorge BlancoRan BlekhmanMark BlennerJoseph M. BlissLouis-Marie BobayThomas A. BobikJames BoedickerAlexandria B. BoehmNicholas Andrew BokulichAndrea BonettiJames L. BonoBatbileg BorErik BorchertAndrew M. BormanJames BornemanThomas C. G. BoschJoseph M. BosilevacMichael J. BouchardYann F. BoucherPatrick H. BradleyEefjan BreukinkAmanda J. BrinkworthShaun R. BrinsmadeJuliana Delatorre BronzatoJulie BrothwellTitus BrownLinda BrubakerSpencer A. BruceSebastian BruchmannBryan D. BrysonGregory A. BuckAngus BucklingSilvia BulgheresiLorinda BullingtonLisa BurdetteLiana T. BurghardtMark J. ButtnerL\u00edlian CaesarFeng CaiIsabelle CaldelariPhilip CalderCarole CamarasaShawn R. CampagnaDanielle Elizabeth CampbellJuliana Coutinho CamposAhmet Can BerkyurekHailong CaoJialan CaoYueqing CaoDelphine CapelaMauricio Caraballo RodriguezValerie Jean CarabettaAlessandra CarattoliJuan Pablo C\u00e1rdenasAllison F. CareyRoss P. CarlsonRonan K. CarrollSantiago Castillo-Ram\u00edrezJuan Castro-SeverynJorge CervantesKok Gan ChanSiu H. J. ChanTrevor C. CharlesBenoit ChassaingSom S. ChatterjeeAdit ChaudharyMax Chavarr\u00edaCongying ChenDing-Qiang ChenFeng ChenGao ChenLiang ChenLianmin ChenShaohua ChenStanley H. ChenYin ChenYongliang ChenCaroline Ch\u00e9nardShu ChengAlexandra ChiaveriniJungil ChoiLon M. ChubizNico J. ClaassensJan ClaesenDennis ClaessenGerard ClarkeThomas ClavelSarah A. ClockMara L. C. CloutierShannon R. ColemanJustine CollierEric CollinsCharles ColuzziAndre M. ComeauFabio CominelliLaurie E. ComstockTerrance G. CooperReilly O. CooperFernando CorralesDoug CossarLauren CowleyKatharine CoyteSean CrossonElena CrottiDavid E. CrowleyNyssa CullinChris CurtinRoy CurtissLeah CuthbertsonWeronika CzabanStefania DaghinoYang DaiBenjamin DainatAlex DajkovicJennifer L. DaleGautam DantasSophie E. DarchPromi DasRaunak Kumar DasSudip DasMark R. DaviesEllen DecaesteckerHidde de JongRonnie de JongeJohn P. DekkerRosa del CampoDiego de MendozaXianding DengXiangyu DengXin DengYijie DengLaura De NiesNicole J. De NiscoDaniel P. DepledgeDhwani DesaiLes DethlefsenTravis J. De WolfeEric D\u00e9zielAvantika DhabariaNeha DhasmanaJuan Diaz-ColungaFabrizio Di CaprioGregory J. DickJoseph DiDonatoChristian DienerStephen P. DiggleKimberly Anne Dill-McFarlandTatiana DimitriuRay DixonUlrich DobrindtYohei DoiStephen K. DolanSara DominguesJustin J. DonatoSharon DonovanCharles J. DormanGavin M. DouglasMarc DroletChao DuYabing DuanBreck A. DuerkopSe\u00e1na DugganJack DunkleAnne DuplouyMatt DurrantSanjucta DuttaRachel J. DuttonM\u00e1ria D\u017eunkov\u00e1William D. EatonTom EdlindCollin EdwardsMartinique Lefevre EdwardsMarie A. ElliotJoshua ElmoreEmiley Ansley Eloe-FadroshMelinda Anne EngevikChristoph EnglTobias EnglBrendan EpsteinAna V. Espinel-IngroffCarmen Espinosa-GongoraAndreia B. EstrelaYanhua FanRongxiang FangWeiguo FangYufeng FangS\u00e9amus FanningSara FedericiVictor FedorenkoConor FeehilyFengqin FengHong FengPinghui FengGabriel da Rocha FernandesJuan Carlos Gutierrez FernandezMarine FeyereisenNoah FiererReinhard FischerRoss FitzgeraldKyle FloydFatima FoflonkerMarco FondiHerrison FontanaGad FrankelPatrick FrantomPaul FraserAlessandra FrauSteven FreseMaximilian FreyMichael W. FriedrichVille FrimanJulia FrunzkeYunhe FuChikara FurusawaGiovanni GalloMichael G. GanzleCheng GaoXiaofeng GaoDavid Garcia-CallejasMelanie GareauJunkal GarmendiaMatthew J. GebertJennifer Geddes-McalisterJohn A. GerltGisa GeroldMonica L. GerthLandon John GetzRohit GhaiMurad GhanimRaad GharaibehPartho GhoshScott Michael GiffordConcha GilMicaela GiorginiMathieu GissotStefanie P. GlaeserAnum GlasgowLaura GlendinningErin S. GloagGregory B. GloorAlain P. GobertMatthew Robert GoddardDaryl GohlJoanna B. GoldbergJoao Carlos Gomes-NetoLaura G\u00f3mez-ConsarnauAngela Gomez-SimmondsYanhai GongCesar Raul Gonzalez-EsquerUri GophnaTobias GorisAndrew GorringeJeffrey M. GrabowskiDavid E. GrahamTodd M. GrecoChris GreeningGwen G. GreletJoel S. GriffittsIn\u00eas Ramos GriloGhjuvan GrimaudTed GrunbergAlexander Gr\u00fcnbergerDanxia GuHaiwei GuSteinn GudmundssonRoberto M. C. GuedesCaitriona M. GuinaneXianwu GuoTony GutierrezAndras GyorgyMaria HadjifrangiskouTatsuro HagiAndrea HahnVanessa L. HaleAlex HallRuth M. HallKambiz HamadaniTrinity L. HamiltonThomas H. HamptonMei-Ling HanNabil HannaGeoffrey D. HanniganPamela HansonWilliam R. HarcombeJessica HardinChristopher HarmerXavier HarrisonMark E. HartChristiane Hassenr\u00fcckGraham F. HatfullRoland HatzenpichlerAlyse HawleySusu HeYan HeAziz HeddiJussi HeinonsaloKlaas J. HellingwerfChris HenryMichael A. HensonMelissa M. Herbst-KralovetzDaniel Philipp Ralf HerlemannMark T. HernandezAnder Hern\u00e1ndez Pl\u00e1garoRobert L. HettichPaul G. 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McHughMichael J. McInerneyJames B. McKinlayMichael J. McLarenJoel McManusRainer U. MeckenstockGregory L. MedlockMaliheh MehrshadLuis C. MejiaMelissa MelbySilvia MelgarBrett L. MellbyeAlessio MengoniGuillaume M\u00e9ricNoha M. MesbahSt\u00e9phane MesnageElla M. MeumannGeorge MichailFiras S. MidaniPeter E. MidfordJoseph R. MihaljevicJeremiah J. MinichBiswapriya Biswavas MisraRajeev MisraRicha MisraDouglas A. MitchellMegan C. MladinichJennifer M. MobberleyLuke A. MoeAmin MohamedYassene MohammedOmkar Satyavan MohiteMashkoor MohsinWendy MokL\u00e1zaro MolinaDenise MonackIsha MongaJonathan M. MonkDouglas MonteiroOlimpio MonteroMatthew D. MooreJames J. MoranChristopher E. MorganMegan MorrisMark MorrisonJamie MortonLuis Mota-BravoJochen A. MuellerRyan Sean MuellerMandy MullerCatherine Mulli\u00e9Alise MuokGemma G. MurrayMario E. MuscarellaJillian MyersCarey D. NadellNiranjan NagarajanDrew R. NanniniAdrienne B. NarrowePayman NasrStephen NayfachDaniel NaylorBrittany NeedhamJeniel E. NettMeina Neumann-SchaalJoshua Patrick Mark NewsonChristopher NicchittaGraeme William NicolPierre NicolasVincent NietoPablo Ivan NikelBen NiuQiuhong NiuFranklin NobregaSamuel-Philip NobsTrent NorthenSpencer V. NyholmAnna O'BrienConor P. O'ByrneKazuhiro OgaiDele OgunremiYasuo OhnishiShigeaki OhnoNaoko OhtaniAnil OjhaAndrew OleinikovAngela OliverioChristine OlsonMarc OngenaMei OoiTodd OsmundsonOrla O'SullivanHiroshi OtaniRonan F. O'TooleMichael OttoHong-Yu OuMarc OuelletteJeremy George OwenEgon Anderson OzerMartin PabstCara T. PagerMelinda PaholcsekSepideh PakpourMagnus PalmbladRobert J. PalmerBernhard O. PalssonMeichen PanTansol ParkJohn ParkinsonJulio Parra-FloresAnutthaman ParthasarathyLaila Pamela Partida-MartinezSally R. PartridgeAdrian PaskeyEdoardo PasolliAlessandro PasseraKiran PatilMichael PatnodeAndrew C. PawlowskiJennifer L. PechalItsik Pe'erDesheng PeiRudy PelicaenDale A. PelletierXuanxian PengJakob PernthalerJames PetersScott PetersonVanessa V. PhelanBrett E. PickettGiovanni PilloniJarone PinhassiAmeet J. PintoJohann PitoutPaul J. PlummerMircea PodarGeorg PohnertLaurent PoirelSuchawan PornsukaromAnais PotronDominic Poulin-LapradeAkbar Adjie PratamaSusan PrescottErin P. PriceMorgan N. PriceSambhawa PriyaAlexander J. ProbstDaniele ProvenzanoQiang PuAlexandra E. PurdyFahd QadirXiaopeng QiZhe-Xue QuanAlisha QuandtKenjiro Wake QuidesRalf RabusMark RadosevichMalini RajanMirjana Rajili\u0107-Stojanovi\u0107Gordon RamageKelly RamirezDipak RamjiJayasimha RaoDavid RaskoSimon RasmussenDaniel RathThomas RatteiAlison RavenscraftKasie RaymannTimothy D. ReadShawna ReedPierre RenaultJose M. RequenaHenry ReyerAlejandro ReyesPhilip J. RichardsJason M. RidlonDaniel Rios GarzaHenrik Munch RoagerSteve RobbinsLeah RobertsSerina L. RobinsonAmalia RocaJennifer RoccaGuillermo RodrigoMarcio RodriguesLuis RodriguezEduardo Rodr\u00edguez-Rom\u00e1nDavid RomeroMarilyn J. RoossinckJason W. RoschFederico RosconiBenjamin RossDaniel E. RossOmar RossiHannes L. RostChristopher RotaDaniel R\u00f6thSimon RouxDenis RoySyamal RoyDaniel RozenPeter RubbensJeffrey RudolfMarc RuitenbergSteffen RuppFracesco RussoSangryeol RyuZakee L. SabreeRajib SahaJason W. SahlSakuntala SaijaiElisa SalvettiNadia SampaioJohn C. SamuelsonNicholas SandovalRobert A. SanfordDominique SanglardAlvaro San-Mil\u00e1nAlyson E. SantoroChristian Santos-Medell\u00ednGuillaume SarrabayrouseJimmy H. SawTomoo SawabeGary SawersMatthew J. ScarboroughJoy ScariaJoseph SchachererLennart Schada von BorzyskowskiIsabelle SchalkDirk-Jan ScheffersBernhard H. SchinkGuy SchleyerPatrick D. SchlossMarian L. SchmidtThomas M. SchmidtDirkjan SchokkerMichelle SchornBenjamin Luke SchulzEgbert SchwartzHannah Doris SchweitzerVera SchwierzeckPeter SeboLeopoldo SegalJana SeifertRyan F. SeipkeYuji SekiguchiCharlie SetoLuca SettanniEmmanuel SeveriMohammad R. SeyedsayamdostLauren Marie SeylerMichael ShafferMaulin P. ShahYatrik ShahFergus ShanahanXihui ShenPaul SheridanJunling ShiAndrey N. ShkoporovAmanda ShoreNatalia ShulzhenkoSeyed Davar SiadatNikolai SiemensSusanne SieversCynthia B. SilveiraAmit SinghAnoop SinghBaneshwar SinghRamandeep SinghNaresh SinghalSonia SinghalEvan SkowronskiTimofey SkvortsovLloyd M. SmithMelvyn SmithPaul SmithThomas E. SmithScott D. SobyChristian SohlenkampBokai SongHyun-Seob SongJiuzhou SongJustin L. SonnenburgUtkarsh SoodLauren SpeareStephen SpiroSanjeeva SrivastavaLee F. StanishFabian StaubachB\u00e4rbel StecherTodd R. SteckBruno StefanonEike J. SteinigUlrich StelzlBo Maxwell StevensBradley S. StevensonAdrie J. SteynPaul StodghillSarah StraussWolfgang R. StreitJian Qiang SuXiaoquan SuWoo Jun SulSnorre SulheimChaomin SunHongzhe SunHui-Zeng SunQiang SunShan SunWenxian SunXiaolun SunPer SunnerhagenMaxim SvetlovAustin D. SwaffordBryan SwingleJason B. SylvanYinjie J. TangWindy TannerGerald W. TannockXuanyu TaoAzuma TaokaIlma TapioCormac Thomas TaylorJan TebbenHerve TettelinChristoph A. ThaissAndrew Maltez ThomasJulie A. ThomasSunnie R. ThompsonCasper ThorupGuo-Bao TianXiaojun TianYun TianMarius Belmondo TinchoKara A. TinkerDirk TischlerAndrzej TkaczJeffery K. TomberlinParizad Torabi-PariziDieter M. TourlousseMaxime TourteSachia Jo TravingGena D. TribbleDenis TrubitsynGareth TrublStephen Kwok-Wing TsuiJennifer Anne TullmanBenjamin John TullyChristine Y. TurennePeter J. TurnbaughCaroline TurnerElhanan TzipilevichDaniel UdwaryJuan A. UgaldeRaphael ValdiviaYo-Ann Valez JustinianoEthan Van ArnamFran\u00e7oise Van BambekeJan Roelof van der MeerKaren Jane VanderwolfJames L. Van EttenRob Van HoudtJolanda van LeeuwenWillem Van SchaikSuzan Pantaroto VasconcellosDelyana Peteva VasilevaNic M. VegaSandeep VenkataramVittorio VenturiN. VerdileAjit VikramValeska Villegas-EscobarAntony T. VincentUwe VoelkerAngelina VolkovaAurele VuilleminJoseph Thomas WadeMaggie R. WagnerIrene Wagner-DoblerLevi WaldronDavid Ian WalkerWolfgang WanekChengshu WangHaixia WangJinfeng WangJoyce WangJunjun WangPandeng WangQiyao WangWeilan WangXifeng WangXueshan WangYue WangYongbao WangIndu WarrierAlex D. WashburneTilmann WeberZhaojun WeiVirginia WeisAlexandra J. WeisbergJake WeissmanRoy D. WelchJia WenI. WheeldonNicole WheelerRichard Allen White IIISivaramesh WigneshwerarajRoland Conrad WilhelmMichael WilkinsJulia WillettAmy D. WillisTomasz WilmanskiJennifer R. WilsonJennifer H. Wilson-WelderAngela WitteAlan J. WolfeSul Woo JulJonathan WrenGerard D. WrightRobyn J. WrightHao WuRuonan WuYu-Wei WuSander WuytsClaudia WylezichKristine M. WyliePeng XingJinbo XiongWei XiongLibin XuPing XuZhenjiang Zech XuKatherine XueTakuji YamadaMasayuki YamamotoQingyun YanTao YanXianghua YanBaowei YangChunfu YangJinkui YangJun YangZhaomin YangYu-Feng YaoM.-N. Frances YapRobert YarchoanEmily Yates-DoerrJung-Yong YehHuabing YinShibu YoosephLoubna YoussarEizadora T. YuGuangtao YuJing YuanMengting Maggie YuanCarlos R. Z\u00e1rate-Blad\u00e9sRaz ZarivachAhmed ZayedDavid ZeeviAnna C. ZemkeLauren A. ZenewiczKarsten ZenglerAngela ZhangGuangliang ZhangJiachao ZhangJian ZhangMingzi ZhangPei ZhangRong ZhangRuifu ZhangXiao-Hua ZhangYongjun ZhangZhengguang ZhangKun ZhaoYingming ZhaoJun ZhengJusheng ZhengShijun J. ZhengYong ZhengHaokui ZhouJianhua ZhouZhi-Gang ZhouBaoli ZhuWenhan ZhuYang ZhuYong-Zhang ZhuRyan M. ZielsNadine ZiemertMichael ZimmermannZhiyong ZongJackie ZorzJasenka ZubcevicMartin ZwanzigLast year, I hoped that we were moving toward the end of the pandemic. I was wrong. Despite the availability of free vaccines that have demonstrated remarkable ability to reduce transmission and serious illness, vaccine rates are still woefully low. It seems that many of our fellow global citizens (that have access to vaccines) have rejected the evidence presented, preferring to put themselves, family members, and the wider community at risk of serious illness and death. However, this apparent rejection of science should not reduce our resolve to provide rigorous review of our peers\u2019 work. Scientific discovery works only because we as a community are willing to assess its accuracy and validity. Providing this service free of charge is an important part of reducing perceived bias. That being said, it is a major time commitment, and every one of us that gives up our limited time to support peer review does so with the understanding that our assessments are important in ensuring that unfounded, nonrigorous results and conclusions are not disseminated widely. The suggestion that we should abandon peer review and let postpublication review sort through the rigor of new science does not account for the new world in which we live. People will use \u201cpublished\u201d research to justify their perspectives, and so the propagation of poorly designed studies and inaccurate findings will further the erosion of the public\u2019s \u201cfaith\u201d in science. Peer review has never been more important than it is now, and the"} +{"text": "PLoS Medicine, volume 2, issue 11: DOI:10.1371/journal.pmed.0020405Inhttp://unstats.un.org/unsd/mi/techgroup/January2005/Series%20update%20status%20query_FC.xls. Accessed 13 October 2005] was publicly accessible at the time of publication of Amir Attaran's article, but is no longer publicly accessible. This file can now be found asReference 3 [United Nations (2005) Data availability analysis. New York: United Nations. Available:Dataset S1(41.5 KB XLS).Click here for additional data file."} +{"text": "PLoS Medicine, volume 2, issue 10: DOI:10.1371/journal.pmed.0020318Inhttp://unstats.un.org/unsd/mi/techgroup/Sept2004/message_to_inter_agency_mdg.pdf. Accessed 1 August 2005] was publicly accessible at the time of publication of Amir Attaran's article, but is no longer publicly accessible. This file can now be found asReference 3 [Deputy Secretary General United Nations (2004) Message to the inter-agency and expert meeting on MDG indicators Geneva 29 September\u20131 October 2004. New York: United Nations. Available:Text S1(44.5 KB DOC).Click here for additional data file."} +{"text": "PLoS Pathogens, volume 3, issue 1: doi: 10.1371/journal.ppat.0030001In The third author's name was incorrectly listed as Claire J. Hoving. The correct name is J. Claire Hoving."} +{"text": "PLoS Computational Biology, volume 3, issue 3: doi: 10.1371/journal.pcbi.0030053In An author's name was misspelled as Fransizka Michor and should read:Franziska Michor"} +{"text": "PLoS Biology, volume 5, issue 5: doi: 10.1371/journal.pbio.0050097In The ninth author's name was incorrectly given as Janet Thonton; it should be Janet Thornton."} +{"text": "PLoS Computational Biology, volume 1, issue:DOI: 10.1371/journal.pcbi.0010079In in was omitted from Equation 6. The correct equation is as shown below:An"} +{"text": "PLoS Medicine, volume 3, issue 3: DOI:10.1371/journal.pmed.0030167Inhttp://ottawagroup.ohri.ca/signatories.html.The URL provided for the Ottawa Group was incorrect. It should be"} +{"text": "Environ Health Perspect 115:1087\u20131093 (2007)], the corresponding author\u2019s address is incorrect. The correct address is S.G. O\u2019Neil, 1071 Blue Hill Ave., Milton, MA 02186. Telephone: 617-333-0500. E-mail:soneil0905@curry.eduIn the article by O\u2019Neil [EHP regrets the error."} +{"text": "PLoS Genetics, volume 2, issue 12: doi:10.1371/journal.pgen.0020205In http://bioinfo.lifl.fr/yass/yass.php.In the Materials and Methods, the URL to the Yass online program was incorrectly listed. The correct URL is"} +{"text": "PLoS Computational Biology, volume 3, issue 6: 10.1371/journal.pcbi.0030107In In the subsection \"Evolutionary conservation of modules and proteins\" of the Materials and Methods section, a link was incomplete. The correct link reads:http://rd.plos.org/10.1371_journal.pcbi.0030107_01_0"} +{"text": "PLoS Genetics, volume 3, issue 5: 10.1371/journal.pgen.0030085In Mr. Eric L. Van Nostrand's name was incorrectly listed in the online citation as Nostrand ELV. The correct citation is:10.1371/journal.pgen.0030085.Yeo GW, Van Nostrand EL, Liang TY (2007) Discovery and Analysis of Evolutionarily Conserved Intronic Splicing Regulatory Elements. PLoS Genet 3(5): e85 doi:"} +{"text": "PLoS Pathogens, volume 3, issue 3: doi: 10.1371/journal.ppat.0030034In The seventh author's name was incorrectly listed as Gilean A. V. McVean. The correct name is Gil McVean."} +{"text": "DOI: 10.1371/journal.pcbi.0020013PLoS Computational Biology, volume 2, issue 2:In http://cssb.biology.gatech.edu/skolnick/files/gpcr/gpcr.html.The URL provided for the GPCR model database in the published article is no longer active. The database is now located at"} +{"text": "E. coli in humans in the Netherlands. PLoS ONE 13(3): e0193834. https://doi.org/10.1371/journal.pone.0193834The fifth author\u2019s name is incorrect. The correct name is: Cornelia C. H. Wielders. The correct citation is: Teunis PFM, Evers EG, Hengeveld PD, Dierikx CM, Wielders CCH, van Duijkeren E (2018) Time to acquire and lose carriership of ESBL/pAmpC producing"} +{"text": "Present: Due to an error made during the assembly of Figure Corrected: Correct Figure 11709-22. doi: 10.18632/oncotarget.2606.Original article: Oncotarget. 2014; 5(22):"} +{"text": "R. Soc. open sci.4, 171085. (Published 6 December 2017). (doi:10.1098/rsos.171085)http://rsos.royalsocietypublishing.org/content/4/12/171085.An author name was given wrongly in reference [17] of The author's name should be Hooper R and not Holder R, as presently written.The corrected reference in full is:p-value depends only weakly on statistical power in realistic situations. J. Clin. Epidemiol. 62, e1242\u2013e1247. (doi:10.1016/j.jclinepi.2009.02.004)Hooper R. 2009 The Bayesian interpretation of a"} +{"text": "PRESENT: Due to an error during production, Figures .Correct: Figures 830-43. doi: 10.18632/oncotarget.971.Original article: Oncotarget. 2013; 4(6):"} +{"text": "Richard Wiseman, Adrian M. Owen. Turning the Other Lobe: Directional Biases in Brain Diagrams. i-Perception. 8(3):1\u20134.The authors regret that the labels on"} +{"text": "The Data Availability statement for this paper is incorrect. The correct statement is: All relevant data are available at the following locations:https://doi.org/10.6084/m9.figshare.4730053.v1Golgi staining: https://doi.org/10.6084/m9.figshare.4730089.v1Hippocampal primary cultures (spine density): https://doi.org/10.6084/m9.figshare.4730038.v1qPCR Tspan6 KO mice: https://doi.org/10.6084/m9.figshare.4730026.v1RNA scope images: https://doi.org/10.6084/m9.figshare.4730146.v1Electrophysiological recordings: https://doi.org/10.6084/m9.figshare.4730149.v1Behavioral results: https://doi.org/10.6084/m9.figshare.4730164.v1Surface GluA1 expression in hippocampal primary neurons: https://doi.org/10.6084/m9.figshare.4730182.v1Western blot hippocampal synaptosomes: https://doi.org/10.6084/m9.figshare.4730194.v1Western blot hippocampal homogenates:"} +{"text": "The Chief Editors and the Editorial Board thank the reviewers' listed below for their contribution and time consuming efforts in refereeing papers submitted to 3 Biotech in 2015. Their timely response and critical comments have not only assisted us in the selection of high-quality scientific work, but their critique is also appreciated by authors as it frequently assists them to improve the style and content of their publications.Nayera A.M. AbdelwahedMar\u00eda Abad-GrauNaglaa AbdallahNadia AbdEl-NasserTapan K. AdhyaDeepti AgrawalBasir AhmadAta AkcilMohd. AkhtarAyodele AlaiyaSahal Al-HajojFatimah AlhamlanRabih Al-KaysiSulaiman Al-MayoufGundi Vijay AnandProf. Reshetilov AnatolyAnees AnsariMohammad AnsariRaheel AnwarAbdolreza ArdeshirylajimiEhtasham ArifMohammad AsadollahiUtku AvciMd. Rabiul AwualYuxiang BaiChunyu BaiN.K. BainsalaM.D. BalakumaranH.S. BalyanIndrani BanerjeeDebdulal BanerjeeHussaina BanuAndrew BattleP.D. BelurSubhash BhardwajPankaj M. BhattDurga BhattChanghao BiS. BinduRajib BiswasYannick BombleAbhijeet BoroleCraig BuntSiddhardha Busi SiddhardhaTao CaiZhen CaiMingfeng CaoAur\u00e9lie C\u00e9bronSonia ChadhaSusanta ChakrabortyAnindya ChandaNidhee ChaudharyS.F. ChenJinchun ChenHairong ChengJoshua ChouKathrine ChristensenHanna DahmP.K. DasAshis DasSurajit DasShailesh DaveAchlesh DavereyJan De RiekApurba DeyD. DhanasekaranNicolas DoucetJoao DuarteKheireddine El-BoubbouNadia El-GamalAhmed ElsharkawySebnem ErenlerLeticia EstevinhoThaddeus EzejiAnbin EzhilanE. Fernandez-GarciaDaniela GaborShowkat GanieYuhua GaoDifeng GaoRomain GautierK. Gawel-BebenXumeng GeSveta GerdesJan GeunsM. GhaediParvatam GiridharAnanthan GnanakkanVenkateswara GogineniT. GopalakrishnaSubramaniam GopalakrishnanBhattiprolu Govinda RaoShipra GuptaDorin GuptaVandana GuptaVijai Kumar GuptaVijai GuptaBaskar GurunathanAshok HadapadSubhashish HaldarYiejun HanZhangying HaoVinayaka HegdeKhan HekmatyarCornelia HooperShankar HosmaniZhiyong HuangK. Hari KrishnanSundeep JaglanRachna JainHasnain JavedR. JayamadhuriVenkatesh JelliSujee JeyapalinaJean JoseChaitanya G. JoshiTarek KabilC.C.N. KhobragadeD. KaladharRajwant KaliaChristos KannasPrasad KaparajuB. KapdnisAfsin KayaKumaraswamy KenchappaR. KeerthiDerek KennedyRavi KesariNeda KeyhaninejadRafeiza KhanSardar KhanThida Win KhinMoon-Soo KimSoo_Ki KimUday KishoreTimo KorhonenAnil KotasthaneAnkita KothariFatemeh KouhkanAnagha KrishnanLakshminarasimhan KrishnaswamyRamesh KuhadPankaj KumarSuresh KumarDr. Narendra KumarSaravanah KumarAaron Alfred KwaasiPark KyeungJinwook LeeSlawomir LewickiS.N. LewisDemao LiZhongyi LiJinshan LiY. LiZhenglong LiYuping LinS.J. LinLifeng LiuTao LiuJin LiuJun LiuXiaowei LiuH. LiuLong LiuJos\u00e9 Pedro Lopes FariaXavier LouisS. LutsenkoThomas LutzHongwu MaDatta MadamwarRajat MahajanS. Dharne MaheshHamid MalekiAnushree MalikK. MallikarjunaNazim MamedovJose ManchenoMaegala ManiyamAshwini MathurSayaji MehetreBoris MinaevPankaj MishraGiovanni MitaM.K. RajeshBiplob ModakReda MoghaiebSanaa MohamedMaysa MoharamThanaa MohdalyDejana MokranjacHossain MondalSukanta MondalPoulomi MukherjeeArup Kumar MukherjeeSuprabhat MukherjeeSuparna MukherjiAkhtar NadhmanAmbarish NagManjunath NaikJyothisha NairSavithri NambeesanBala NambisanRenu NandakumarIsali NantesPradeep NegiHelena NevalainenSujogya PandaJanmejay PandeyPranay PankajEmmanuel PapamichaelM.V. ParakhiaRamakrishnan ParthasarathiTrupti PatelRanjana PathaniaKishore PatilChristina PayneI. PeinadoYanfeng PengAngelo PeraltaCarlo PiermarocchiOnruthai PinyakongVimal PrajapatiA.P. PratapBharath PrithivirajT. PullaiahHong QingBabak RabieiB. Rajasekhar ReddyK. RaghavendraHamidur RahamanSudhir RaiNishant RaiNaushad RaisS. RajagopalAlex Selvanayagam RajangamBernard Rajeev SWVrinda RamakrishnanB.V. RamanV. RangaswamyA.R. RaoRino RappuoliMamoon RashidArthi RathinasabapathyGourav RathoreRajesh RautPrasun RayManju RayMehdi Razzaghi-abyanehGopal ReddyK. RekhaSatyanarayana RentalaKedar RokadeMainak RoySeunghyun RyuA. SabuJitendra SainiS.S. SandhuMaddirevulla SateeshP. Suresh kumarS. SatheeshKannusamy SathyaseelanTulasi SatyanarayanaIndu SawantWalter SchmidtIvan SchusterSamuel SeaverG. Seghal KiranFlavio SeixasJoseph SelvinAn Seong Soo AHanan ShaabanShafinaz ShahirSitansh SharmaBirinchi SharmaNitya SharmaR. ShashidharIng-Lung ShihP. ShilpkarWenqing ShuiPratyoosh ShuklaSwapna SimonRandhir SinghAbhishek SinghKusum SolankiAndong SongCunjiang SongHui SongMaria SotoValentina SpanicJanet SprentBharati SrinivasanS. SriramManoj SrivastavaAlexander Steinb\u00fcchelM. Subhosh ChandraMayavan SubramanianKondeti SubramanyamRajeev SukumaranFaheem Sultan MohammadGeeta SumbaliYuanxia SunP.N. SunilkumarLaura A. SvetazTamer TamerLi TanGeok Hun TanShuangyan TangFei TaoHamsa TayebValentino Te\u2019oChao-guang TianAdeline TIngNathan TintleJoe TiralongoMontserrat TobajasM. TohidfarIrena Trbojevi\u0107 Akma\u010di\u0107Minh TriTimir TripathyRan TuP. Uma Maheswari DeviNaryana UpadhyayaS.V.N. VijayendraO.P. VermaSalwa WahshDongping WangZhen WangYi WangHaiyan WangChonglong WangThomas WestQingyu WuYi-Rui WuQiaqing WuHaixia XiaoJianmin XingZhenghong XuZhuofei XuS. XuVinod YadavJohn YarbroughG\u00f6ksungur YektaDr. Sailu YellaboinaRachel YesudasanDoctor Xiaoxing YinYanbin YinBo YuXiaochen YuYongbo YuanYouxi YuanHaggag ZainLila ZareiY.L. ZhangYanping ZhangGuimin ZhangDongyuan ZhangLing ZhangM.W. ZhangY. ZhangYifeng ZhangDawei ZhangDong ZhangQ. ZhangYingying ZhengChe Zhen-mingCheng ZhouKun Zhu"} +{"text": "The Editor has retracted this article due to sNanoscale Res. Lett. 11:462; first published 18 October 2016. Huang Z-Optics Commun. 383:1-5; first published 30 August 2016. Huang Z-"} +{"text": "Present: A reference for this paper was accidentally omitted from the article.Correct: The missing reference appears below.15. Al-haidari AA, Syk I, Jirstr\u00f6m K, Thorlacius H. CCR4 mediates CCL17(TARC)-induced migration of human colon cancer cells via RhoA/Rho-kinase signaling. Int J Colorectal Dis. 2013; 28:1479-1487.10.18632/oncotarget.10256Original article: Oncotarget. 2016; 7:47637-47649. doi:"} +{"text": "This article has been corrected: Dr. Serban San-Maria was added to the author list.The authors sincerely apologize for this oversight.41363-41379. https://doi.org/10.18632/oncotarget.9133Original article: Oncotarget. 2016; 7:"} +{"text": "The original version of this article unfortunThe corrected Reference 19 is given below.https://www.blacknote.com/wp-content/uploads/2017/12/LDDIStatement-2.pdf.19. Institute for Children\u2019s Environmental Health."} +{"text": "This article has been corrected: The online version of figure 3 has been corrected:9766-9775. https://doi.org/10.18632/oncotarget.23919Original article: Oncotarget. 2018; 9:"} +{"text": "In the original publication have twoIncorrect:Zoe Z. MarshmanPhilip P. BensonCorrect:Zoe MarshmanPhilip Benson"} +{"text": "Present: The funding acknowledgements were omitted from the original paper.Correct: The proper funding acknowledgements are given below.doi: 10.18632/oncotarget.8862Original article: Oncotarget. 2016; 7:45094-45111."} +{"text": "Present: There is a duplication of images within Figure 4BCorrect: The proper figure images are shown below. The authors sincerely apologize for this error.37966-37978. doi: 10.18632/oncotarget.9274Original article: Oncotarget. 2016; 7:"} +{"text": "In the original publication two Goog3. Sherstyuk VP, Sarapulova OO, Shvalagin VV. Luminescent hybrid nanocomposites and prospects of molecular and nanophotonic systems in modern packaging and printing. Repino, St. Peterburg, Russia: Book of Abstracts of the 3-International Symposium \u201cMolecular Photonics\u201d; 2012. p. 79.https://scholar.google.com.ua/scholar?hl=uk&q=Luminescent+hybrid+nanocomposites+and+prospects+of+molecular+and+nanophotonic+systems+in+modern+packaging+and+printing&btnG=http://onlinereg.ru/mph2012/Molecular_photonics2012_Abstracts.pdf#page=7913. Sarapulova O Kyrychok T, Sherstiuk V, Orlov A. Modern printing technologies for micro- and nanoelectronics. Proceedings of IEEE XXXIII International Scientific Conference Electronics and Nanotechnology (ELNANO) April 16\u201319, 2013 Kyiv, Ukraine 2013: 151\u2013155.https://scholar.google.com.ua/scholar?q=Sarapulova+O+Kyrychok+T%2C+Sherstiuk+V%2C+Orlov+A.+Modern+printing+technologies+for+micro-+and+nanoelectronics.&btnG=&hl=uk&as_sdt=0%2C5"} +{"text": "Correction to: Translational Psychiatry (2017) 7, e1011; doi:10.1038/tp.2016.281; published online 24 January 2017The 14th author's name was presented incorrectly. The correct listing is C Pantelis."} +{"text": "In the original publication of this article , the accThe correct details of the NCBI accession numbers can be found below:Availability of data and materialsThe genomic sequencing data and assembled and annotated genomes are deposited at NCBI under bioproject accession numbers PRJNA304627 (K. pastoris), PRJNA304977 (K. phaffii wildtype), and PRJNA304976 (K. phaffii GS115). RNA-seq data are deposited at NCBI under the bioproject accession numbers PRJNA311606.In addition to this, please find the direct links to the data below:Transcriptome study:http://www.ncbi.nlm.nih.gov/bioproject/PRJNA311606http://www.ncbi.nlm.nih.gov/sra?linkname=bioproject_sra_all&from_uid=311606Genome assemblies:http://www.ncbi.nlm.nih.gov/bioproject/PRJNA304627http://www.ncbi.nlm.nih.gov/bioproject/PRJNA304977http://www.ncbi.nlm.nih.gov/bioproject/PRJNA304976"} +{"text": "This article has been corrected: Dr. Fu and Dr. Xu citations have been changed to affiliation 1 and affiliation 5 is now deleted.https://doi.org/10.18632/oncotarget.19382Original article: Oncotarget. 2017; 8:89364-89374."} +{"text": "Phil. Trans. R. Soc. B371, 20150360 (doi:10.1098/rstb.2015.0360)After publication, a mistake was identified in equation (2.7); the denominators in the equation should have a minus sign rather than a plus. The corrected equation is provided here."} +{"text": "This article has been corrected: Dr. Mei Li is now listed as the first correspondence author instead of Dr. Jian Yu in the correspondence section.9280-9292. https://doi.org/10.18632/oncotarget.14063Original article: Oncotarget. 2017; 8:"} +{"text": "Present: Due to an error during figure assembly, the center panel of Figure Corrected: The corrected Figure 9135-49. doi: 10.18632/oncotarget.7035.Original article: Oncotarget. 2016; 7(8):"} +{"text": "Present: The grant information is incomplete.Correct: Additional grant information is shown below.18736-18749. https://doi.org/10.18632/oncotarget.7702Original article: Oncotarget. 2016; 7:GRANT SUPPORTNational Natural Science Foundation of China ."} +{"text": "Jackie E. MaharJackie E. Maharet al.). The article has been corrected online (https://wwwnc.cdc.gov/eid/article/24/1/17-0412_article)."} +{"text": "Present: The currently listed corresponding author, John C. Herr, is deceased. Oncotarget wishes to express our condolences to his family and colleagues.Corrected: The new correspondence author information is as follows.30194-211. doi: 10.18632/oncotarget.4734.Original article: Oncotarget. 2015; 6(30):Correspondence to:Eusebio S. Pires,email:eusebiopires@gmail.com"} +{"text": "There is an error in reference 71. The correct reference is:Drosophila Mushroom Body Neurons. PLoS ONE 4(12): e8392. https://doi.org/10.1371/journal.pone.000839271. Lin S, Huang Y, Lee T (2009) Nuclear Receptor Unfulfilled Regulates Axonal Guidance and Cell Identity of"} +{"text": "This article has been corrected: Dr. Meiqing Lou is included as a co-corresponding author.706-717. https://doi.org/10.18632/oncotarget.23091Original article: Oncotarget. 2018; 9:"} +{"text": "C. elegans. The correct citation is: Xie M, Roy R (2015) The Causative Gene in Chanarin-Dorfman Syndrome Regulates Lipid Droplet Homeostasis in C. elegans. PLoS Genet 11(6): e1005284. doi:10.1371/journal.pgen.1005284In the title of this article, the words \u201cChanarian Dorfman\u201d should read \u201cChanarin-Dorfman\u201d. The correct title is: The Causative Gene in Chanarin-Dorfman Syndrome Regulates Lipid Droplet Homeostasis in"} +{"text": "Journal of Experimental Botany, Vol. 68, No. 15 pp. 4219\u20134232, 2017 doi: 10.1093/jxb/erx233The original published version of this article spelled an author\u2019s name incorrectly. The correct name is as follows: Kresanth Varatharajah."} +{"text": "Present: Due to a proofreading error, one of the author\u2019s names was misspelled as: Nilofer Z. Azad.Correct: The proper spelling is as follows: Nilofer S. Azad. The publishers sincerely apologize for this oversight.25950-61. doi: 10.18632/oncotarget.7436.Original article: Oncotarget. 2016; 7(18):"} +{"text": "Scientific Data 3:160047 doi: 10.1038/sdata.2016.47 (2016); Published 5 July 2016; Updated 13 September 2016Since publication, the authors have additionally deposited these data to figshare to further ensure that the data are easily accessible to all researchers:figsharehttps://dx.doi.org/10.6084/m9.figshare.c.3291368 (2016).Wang, L. & Chen, L."} +{"text": "This article has been corrected: the 5th affiliation citation has been included to Dr. Jiun-Jie Wang along with affiliation 6th.https://doi.org/10.18632/oncotarget.15904Original article: Oncotarget. 2017; 8:62606-62621."} +{"text": "This article has been corrected: Dr. Jian Chen was added to the author list.The authors sincerely apologize for this oversight.640-649. https://doi.org/10.18632/genesandcancer.151.Original article: Genes&Cancer. 2017;8:PMID: 28966725; PMCID: PMC5620009"} +{"text": "Legionella pneumonia 2015EPI-NEWS 42/43, 201626 October 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2043%20-%202016.aspxEnhanced Surveillance of Mycobacterial Infections (ESMI) in Scotland: 2016 tuberculosis annual report for ScotlandHPS Weekly Report, 2016;50(43)25 October 2016, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2016/1643.pdfMycoplasma pneumoniae epidemicEPI-NEWS 41, 201612 October 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2041%20-%202016.aspxMiddle East respiratory coronavirus (MERS-CoV). Monitoring of suspected cases in France, October 2012\u2013 December 2015Bulletin \u00e9pid\u00e9miologique hebdomadaire, 32\u20133311 October 2016, Francehttp://invs.santepubliquefrance.fr/beh/2016/32-33/pdf/2016_32-33.pdfInfluenza activity in mainland France, season 2015\u20132016Bulletin \u00e9pid\u00e9miologique hebdomadaire, 32\u20133311 October 2016, Francehttp://invs.santepubliquefrance.fr/beh/2016/32-33/pdf/2016_32-33.pdfVTEC in Scotland 2015: enhanced surveillance, reference laboratory and clinical reporting dataHPS Weekly Report, 2016;50(42)18 October 2016, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2016/1642.pdfSalmonella trends in humans, farm animals and foodInfectieziekten Bulletin 2016; 27(8)October 2016, the Netherlandshttp://www.rivm.nl/Documenten_en_publicaties/Algemeen_Actueel/Uitgaven/Infectieziekten_Bulletin/Jaargang_27_2016/Oktober_2016/Inhoud_oktober_2016/Trends_in_Salmonella_bij_de_mens_landbouwhuisdieren_en_in_voedselTrichinellosis outbreak after eating wild boar meat in Limburg and Antwerp restaurants in December 2014Vlaams Infectieziektebulletin; 3/2016October 2016, Belgiumhttps://www.zorg-en-gezondheid.be/sites/default/files/atoms/files/2016-3-Trichinellose-outbreak-%20A.forier.pdfInvasive meningococcal disease in Germany 2012 \u2013 2015Epidemiologisches Bulletin 43, 201631 October 2016ehttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2016/Ausgaben/43_16.pdf?__blob=publicationFilThe epidemiology of human papilloma virus in women aged 20\u201365 years living in remote villages in French Guiana: Adapting interventions to the territoryBulletin \u00e9pid\u00e9miologique hebdomadaire, 3418 October 2016, Francehttp://invs.santepubliquefrance.fr/beh/2016/34/pdf/2016_34.pdfCases of pertussis on the rise this autumnFolkh\u00e4lsomyndigheten website12 October 2016, Swedenhttps://www.folkhalsomyndigheten.se/nyheter-och-press/nyhetsarkiv/2016/oktober/fler-fall-av-kikhosta-i-host/Rubella screening of pregnant women by midwivesInfectieziekten Bulletin 2016; 27(8)October 2016, the Netherlandshttp://www.rivm.nl/Documenten_en_publicaties/Algemeen_Actueel/Uitgaven/Infectieziekten_Bulletin/Jaargang_27_2016/Oktober_2016/Inhoud_oktober_2016/Rubellascreening_bij_zwangere_vrouwen_door_verloskundigenHIV infection and AIDS: Quarterly report to 30 June 2016HPS Weekly Report, 2016;50(41)11 October 2016, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2016/1641.pdfZika virus infection in a traveller returning from VietnamEpidemiologisches Bulletin 42, 201624 October 2016https://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2016/Ausgaben/42_16.pdf?__blob=publicationFileInitial Danish experiences with Zika virus DiagnosticsEPI-NEWS 41, 201612 October 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2041%20-%202016.aspxChikungunya and dengue surveillance in mainland France, 2015Bulletin \u00e9pid\u00e9miologique hebdomadaire, 32\u20133311 October 2016, Francehttp://invs.santepubliquefrance.fr/beh/2016/32-33/pdf/2016_32-33.pdfRabies in Belgium, a potential danger? One caseVlaams Infectieziektebulletin; 3/2016October 2016, Belgiumhttps://www.zorg-en-gezondheid.be/sites/default/files/atoms/files/2016-3-Rabies-B-casus-F.Hoefkens.pdfNo urban myth: reception of monkeys with herpes B virus infectionInfectieziekten Bulletin 2016; 27(8)October 2016, the Netherlandshttp://www.rivm.nl/Documenten_en_publicaties/Algemeen_Actueel/Uitgaven/Infectieziekten_Bulletin/Jaargang_27_2016/Oktober_2016/Inhoud_oktober_2016/Geen_broodjeaapverhaal_opvang_van_apen_met_herpes_B_virusinfectie"} +{"text": "Fackler SW, Alexandrakis V, K\u00f6nig D, et al. Combinatorial study of Fe-Co-V hard magnetic thin films. Sci Technol Adv Mater. 2017;18:231\u2014238.http://dx.doi.org/10.1080/14686996.2017.1287520When the above article was first published online, an incorrect version of Figure 5(a) was inadvertently included. The correct figure is shown below.Taylor & Francis apologises for this error."} +{"text": "Present: Due to an error in proofreading, Figures Correct: The proper figures are given below. The author sincerely apologizes for this error.36774-88. doi: 10.18632/oncotarget.4908.Original article: Oncotarget. 2015; 6(34):"} +{"text": "Caenorhabditis elegans. PLoS Genet 12(8): e1006227. https://doi.org/10.1371/journal.pgen.1006227The second author's name is spelled incorrectly. The correct name is: Andreas Rechtsteiner. The correct citation is: Ahn JH, Rechtsteiner A, Strome S, Kelly WG (2016) A Conserved Nuclear Cyclophilin Is Required for Both RNA Polymerase II Elongation and Co-transcriptional Splicing in"} +{"text": "Amphisorus kudakajimensis Using Calcein Acetoxymethyl Ester. PLoS ONE 11(11): e0165844. https://doi.org/10.1371/journal.pone.0165844The last author's name is spelled incorrectly. The correct name is: Takashi Nakamura. The correct citation is: Ohno Y, Fujita K, Toyofuku T, Nakamura T (2016) Cytological Observations of the Large Symbiotic Foraminifer http://www.jst.go.jp/global/english/about.html. The publisher apologizes for the errors.The following information is missing from the Funding section: This work was supported by JST/JICA SATREPS, URL:"} +{"text": "R. Soc. open sci. 4, 160606. (Published 11 January 2017). (doi:10.1098/rsos.160606)This correction is to expand upon on the electronic supplementary material presented in the manuscript.https://dx.doi.org/10.6084/m9.figshare.c.3653213).In the corresponding electronic supplementary material file, additional information about acoustic calculations and data collection has been provided (http://dx.doi.org/10.5061/dryad.s6f4q.Additionally, 10-min sample recordings of both sample sites have been made available via Dryad:"} +{"text": "This article has been corrected: Dr. John Yim was added to the author list.The authors sincerely apologize for this oversight.https://doi.org/10.18632/oncotarget.14476Original article: Oncotarget. 2017; 8:26414-26423."} +{"text": "National Black HIV/AIDS Awareness Day is observed each year on February 7 to emphasize the continuing disproportionate impact of human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) on the U.S. black/African American (black) population.https://www.cdc.gov/hiv/pdf/library/reports/surveillance/cdc-hiv-surveillance-report-2016-vol-28.pdf).In 2014, non-Hispanic blacks represented 12% of the U.S. population (https://www.cdc.gov/features/BlackHIVAIDSAwareness.CDC supports a range of efforts to reduce the risk for acquiring or transmitting HIV infection among blacks. Additional information is available at"} +{"text": "Present: Author contributions were not correctly attributed..Correct: The proper author contributions are shown below.Huang Jian and Shen Fangrong have contributed to this work and are co-first authors.13116-13125. doi: 10.18632/oncotarget.14471Original article: Oncotarget. 2017; 8:"} +{"text": "Present: The images displayed in Figure Correct: The proper figure 84839-84850. doi: 10.18632/oncotarget.13321Original article: Oncotarget. 2016; 7:"} +{"text": "Present: Due to an error in proofreading, a research project number was accidentally omitted from the Acknowledgments list.Correct: The correct numbers are provided below. The authors sincerely apologize for this oversight.33111-24. doi: 10.18632/oncotarget.8880.Original article: Oncotarget. 2016; 7(22):"} +{"text": "Present: The author name \u2018Sung Min Kim\u2019 is spelled incorrectly.Correct: The proper spelling is \u2018Seong Min Kim\u2019.https://doi.org/10.18632/oncotarget.14818Original article: Oncotarget. 2017; 8:16912-16924."} +{"text": "Vol. 112(11): 769-774, 2017.p. 769Phillip Noel Suffysshould read:Philip Noel Suffys"} +{"text": "Present: The grant funding number is incorrect.Correct: The proper number is CMRPD891183. The authors sincerely apologize for this error.https://doi.org/10.18632/oncotarget.10057Original article: Oncotarget. 2016; 7:44047-44061."} +{"text": "EditorThe following manuscript has been retracted from May-June 2015 issue on the request of theauthors who stated that \u201cafter publication, their group found that it was difficultto repeat the results. We believe that there may be some flaws or operational loopholes,hence we would like to retract this paper.\u201d- Retraction in: Pak J Med Sci 2015;31(3):672-677. doi:http://dx.doi.org/10.12669/pjms.313.7170 Link:http://pjms.com.pk/index.php/pjms/article/view/7170"} +{"text": "Streptococcus suis was incorrectly described in the text of Streptococcus suis Serotype 2 Capsule In Vivo . It is a gram-positive bacterium. The article has been corrected online (https://wwwnc.cdc.gov/eid/article/22/10/15-1640_article)."} +{"text": "R. Soc. open sci.4, 170975. (Published 11 October 2017). (doi:10.1098/rsos.170975)Table 1 was presented incorrectly in the published paper. The corrected table is shown below."} +{"text": "This article has been corrected: Dr. Xu citation has been changed to affiliation 1 and affiliation 3 is now deleted.https://doi.org/10.18632/oncotarget.17946Original article: Oncotarget. 2017; 8:68795-68808."} +{"text": "This article has been corrected: Dr. Gabriele Grunig was added to the author list.The authors sincerely apologize for this oversight.35609-35618. https://doi.org/10.18632/oncotarget.16011Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: The correct author name is given below:Jinsong Bianhttps://doi.org/10.18632/oncotarget.23356Original article: Oncotarget. 2018; 9:11414-11426."} +{"text": "Present: The Acknowledgment information is incomplete.Correct: The proper funding acknowledgments are given below.10.18632/oncotarget.2748Original article: Oncotarget. 2015; 6:1605-1617. doi:"} +{"text": "The initials DW appear incorrectly in the Author Contributions. The correct contributions are: Conceptualization: AMBG WFH MAH. Data curation: MBK MAH. Formal analysis: MBK MAH. Investigation: MBK MAH. Methodology: AMBG WFH MAH. Resources: MBK AMBG WFH MAH. Software: MAH. Validation: MBK AMBG WFH MAH. Visualization: MBK. Writing\u2014original draft: MBK. Writing\u2014review & editing: MBK AMBG WFH MAH. The publisher apologizes for the error."} +{"text": "R. Soc. open sci. 171717. (Published Online 14 February 2018). (doi:10.1098/rsos.171717)This correction concerns equation 2.17 and consequently figs."} +{"text": "Present: The originally supplied Figure Correct: The proper Figure 10.18632/oncotarget.5548Original article: Oncotarget. 2015; 6:40053-67. doi:"} +{"text": "There was an error in the Data Summary in the published article. PacBio assembly numbers beginning with CP0119XX should be CP0179XX, as shown below.https://www.ncbi.nlm.nih.gov/nuccore/CP017928; https://www.ncbi.nlm.nih.gov/nuccore/CP017929; https://www.ncbi.nlm.nih.gov/nuccore/CP017930; https://www.ncbi.nlm.nih.gov/nuccore/CP017931; https://www.ncbi.nlm.nih.gov/nuccore/CP017932; https://www.ncbi.nlm.nih.gov/nuccore/CP017933).CAV1015 CP017928\u2013CP017933 .CAV1016 CP017934\u2013CP017937 (The author apologizes for any inconvenience."} +{"text": "Healthcare-associated infections and antibiotic resistanceProperties, frequency and distribution of vancomycin-resistant enterococci (VRE) in Germany - Update 2015/2016Epidemiologisches Bulletin 46, 201716 November 2017, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2017/Ausgaben/46_17.pdf?__blob=publicationFile\u00a0Food- and waterborne diseasesCampylobacter enteritis - risk factors and sources of infection in GermanyEpidemiologisches Bulletin 44, 20172 November 2017, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2017/Ausgaben/44_17.pdf?__blob=publicationFile\u00a0Listeria monocytogenes in the Netherlands, 2016Surveillance of Infectieziekten Bulletin 2017; 28(8)17 October 2017, the Netherlandshttp://www.rivm.nl/Documenten_en_publicaties/Algemeen_Actueel/Uitgaven/Infectieziekten_Bulletin/Jaargang_28_2017/Oktober_2017/Inhoud_oktober_2017/Surveillance_van_Listeria_monocytogenes_in_Nederland_2016\u00a0HepatitidesHepatitis A 2015-2016EPI-NEWS 47, 201722 November 2017, Denmarkhttps://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2047%20-%202017.aspx\u00a0Vaccine-preventable diseasesMeningococcal disease 2016EPI-NEWS 49, 20176 December 2017, Denmarkhttps://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2049%20-%202017.aspx\u00a0Measles outbreak 2017Epi-Insight 2017;18(12)December 2017, Irelandhttp://ndsc.newsweaver.ie/epiinsight/5731j3gwddpimcmkeer4wk?a=2&p=52679956&t=17517804\u00a0A polio-free world?EPI-NEWS 48, 201729 November 2017, Denmarkhttps://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2048%20-%202017.aspx\u00a0Laboratory confirmed cases of measles, mumps and rubella, England: July to September 2017Health Protection Report; 11(42)24 November 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/662499/hpr4217_mmr.pdf\u00a0Special issue: Vaccination of\u00a0young children: Data for\u00a0a\u00a0better understanding of\u00a0public actionBulletin \u00e9pid\u00e9miologique hebdomadaire19 October 2017, Francehttp://invs.santepubliquefrance.fr/Publications-et-outils/BEH-Bulletin-epidemiologique-hebdomadaire/Archives/2017/BEH-hors-serie-Vaccination-des-jeunes-enfants-des-donnees-pour-mieux-comprendre-l-action-publique\u00a0Invasive pneumococcal disease and adherence to pneumococcal vaccination in the childhood vaccination programme 2016EPI-NEWS 41, 201711 October 2017, Denmarkhttps://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2041%20-%202017.aspx\u00a0Respiratory diseases\u00a0Legionnaire\u2019s disease 2016EPI-NEWS 45, 20178 November 2017, Denmarkhttps://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2045%20-%202017.aspx\u00a0Enhanced Surveillance of Mycobacterial Infections (ESMI) in Scotland: 2017 tuberculosis annual report for ScotlandHPS Weekly Report 2017; 51(43)31 October 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1743.pdf\u00a0Influenza activity in France, season 2016-17Bulletin \u00e9pid\u00e9miologique hebdomadaire, 2210 October 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/22/pdf/2017_22.pdf\u00a0Hygiene and prevention practices of respiratory infections during the winter months: results from the 2016 Health Barometer, FranceBulletin \u00e9pid\u00e9miologique hebdomadaire, 2210 October 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/22/pdf/2017_22.pdf\u00a0Perceptions and behaviors of people aged 65 to 75 towards seasonal flu vaccination in France, 2016Bulletin \u00e9pid\u00e9miologique hebdomadaire, 2210 October 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/22/pdf/2017_22.pdf\u00a0Method to increase vaccination coverage of healthcare workers for seasonal influenza in health care institutionsVlaams Infectieziektebulletin; 2/2017October 2017, Belgiumhttps://www.zorg-en-gezondheid.be/sites/default/files/atoms/files/VIB%202017-2_HRES_0.pdf\u00a0Sexually transmitted diseasesSpecial edition: World AIDS Day, December\u00a01, 2017Bulletin \u00e9pid\u00e9miologique hebdomadaire, 29-3028 November 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/29-30/pdf/2017_29-30.pdf\u00a0HIV infection and AIDS: Quarterly report to 30 September 2017HPS Weekly Report 2017; 51(47)28 November 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1747.pdf\u00a0Estimation of the number of new HIV infections and the total number of people living with HIV in GermanyEpidemiologisches Bulletin 47, 201723 November 2017, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2017/Ausgaben/47_17.pdf?__blob=publicationFile\u00a0Shigella notified amongst MSM, June 2017Cluster of cases of multi-drug resistant Epi-Insight 2017;18(11)November 2017, Irelandhttp://ndsc.newsweaver.ie/epiinsight/le4wjue46ls?a=2&p=52547623&t=17517804\u00a0HSE recommends antiretroviral therapy for all people living with HIV attending HIV services in IrelandEpi-Insight 2017;18(10)October 2017, Irelandhttp://ndsc.newsweaver.ie/epiinsight/t8q985ffg1610gkzp9yxn5?a=2&p=52412028&t=17517804\u00a0HIV in Ireland 2016Epi-Insight 2017;18(10)October 2017, Irelandhttp://ndsc.newsweaver.ie/epiinsight/t8q985ffg1610gkzp9yxn5?a=2&p=52412028&t=17517804\u00a0Genital chlamydia and gonorrhoea infection in Scotland: laboratory diagnoses 2007 \u2013 2016HPS Weekly Report 2017; 51(38)26 September 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1738.pdf\u00a0Gonococcal antibiotic surveillance in Scotland (GASS): prevalence, patterns and trends in 2016HPS Weekly Report 2017; 51(38)26 September 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1738.pdf\u00a0Zoonoses and vector-borne diseases Surveillance of human hantavirus infections in metropolitan France, 2012-2016Bulletin \u00e9pid\u00e9miologique hebdomadaire, 2324 October 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/23/pdf/2017_23.pdf\u00a0Zoonotic disease in ScotlandHPS Weekly Report 2017; 51(42)24 October 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1742.pdf\u00a0Infection with TBE virus in Denmark 2013-2016EPI-NEWS 40, 20174 October 2017, Denmarkhttps://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2040%20-%202017.aspx\u00a0Other\u00a0Special edition: Mayotte: epidemiological data for the assessment and prevention of health risksBulletin \u00e9pid\u00e9miologique hebdomadaire, 24-2531 October 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/24-25/pdf/2017_24-25.pdf\u00a0Reportable diseases in the summer of 2017Epi-Ice 10(4), 2017October 2017, Icelandhttps://www.landlaeknir.is/servlet/file/store93/item33182/EPI-ICE_October_2017.pdf\u00a0Outbreaks in the summer of 2017Epi-Ice 10(4), 2017October 2017, Icelandhttps://www.landlaeknir.is/servlet/file/store93/item33182/EPI-ICE_October_2017.pdf"} +{"text": "Present: Figure Correct: The proper figure appears below.10.18632/oncotarget.8111Original article: Oncotarget. 2016; 7:23425-23438. doi:"} +{"text": "This article has been corrected: The 2nd affiliation is added to the author given below:1,2Bou-Yue Peng97153-97164. https://doi.org/10.18632/oncotarget.21315Original article: Oncotarget. 2017; 8:"} +{"text": "PLoS ONE 12(5): e0176821. https://doi.org/10.1371/journal.pone.0176821The second author\u2019s name is incorrect. The correct name is: Channa N. Jayasena. The correct citation is: d\u2019Anglemont de Tassigny X, Jayasena CN, Murphy KG, Dhillo WS, Colledge WH (2017) Mechanistic insights into the more potent effect of KP-54 compared to KP-10"} +{"text": "Volume 8, Issue 5, pages 884\u2010893. First published: 18 March 2014.https://doi.org/10.1016/j.molonc.2014.03.003The Dub3 siRNA oligonucleotide sequence mentioned in this article is incorrect. The correct sequence of the used Dub3 siRNA oligo is GGAGAUCCAAAGGGAAGAAdTdT.The authors apologise for any inconvenience caused by this error."} +{"text": "This article has been corrected: Dr. Sinha has been added in the author list with an equal contribution note along with Dr. Ryan and Dr. Bogan.103182-103206. https://doi.org/10.18632/oncotarget.20892Original article: Oncotarget. 2017; 8:"} +{"text": "Present: Due to a technical error during image processing, an incorrect high resolution version of Figure Corrected: Correct Figure 12156-73. doi: 10.18632/oncotarget.3495.Original article: Oncotarget. 2015 May 20; 6(14):"} +{"text": "The correct name is: Karimollah Hajian-Tilaki. The correct citation is: Omidvar S, Faramarzi M, Hajian-Tilaki K, Nasiri Amiri F (2018) Associations of psychosocial factors with pregnancy healthy life styles. PLoS ONE 13(1): e0191723. https://doi.org/10.2224/sbp.6703. In press.There is an error in reference 30. The correct reference is: Faramarzi M, Pasha H. Psychometric properties of Persian version of Prenatal Distress Questionnaire. Soc Behav Pers. 2018, 45."} +{"text": "Sorghum bicolor (L.) Moench) and related model species. PLoS ONE 13(3): e0192678. https://doi.org/10.1371/journal.pone.0192678The third author's name is spelled incorrectly. The correct name is: Junaid Gamieldien. The correct citation is: Woldesemayat AA, Modise DM, Gamieldien J, Ndimba BK, Christoffels A (2018) Cross-species multiple environmental stress responses: An integrated approach to identify candidate genes for multiple stress tolerance in sorghum ("} +{"text": "This article has been corrected: the title has been corrected.https://doi.org/10.18632/oncotarget.17051Original article: Oncotarget. 2017; 8:41078-41090."} +{"text": "Danio rerio hars gene encodes both cytoplasmic and mitochondrial histidyl-tRNA synthetases. PLoS ONE12(9): e0185317. https://doi.org/10.1371/journal.pone.0185317The third author\u2019s name is spelled incorrectly. The correct name is: Christopher S Francklyn. The correct citation is: Waldron AL, Cahan SH, Francklyn CS, Ebert AM (2017) A single"} +{"text": "While not members of the Board of Editors, invited editors serve an important role in the review process. Invited editors are experts in their fields of research who add an additional level of quality to the review process. An editor may assign a paper to an invited editor when he/she would like to have an additional expert opinion of the reviews or when the subject area falls outside the editor\u2019s primary area of expertise.On behalf of the editors of Dan I. AnderssonMartin Fabian BachmannShweta BansalFernando BaqueroChristopher F. BaslerJoseph Bondy-DenomyArpita BoseMichael J. BrennanJoseph BreseeRoland BroschC. Titus BrownRut Carballido-L\u00f3pezByron CaugheyJean CelliSwaine L. ChenPeter ChienJeff A. ColeVaughn S. CooperBryan R. CullenChristina A. CuomoAndrew J. DarwinRajendar DeoraFloyd E. DewhirstAlain FillouxKlas FlardhKevin FosterVance G. FowlerPhilippe GerardDeanna GibsonN. Louise GlassFIlipa Godoy-VitorinoVernita GordonJeffrey A. GralnickJean GruenbergKaren GuilleminJohn S. GunnDavid L. GutnickLynn E. HancockNancy D. HansonRobert L. HarrisonJeffrey P. HendersonBetsy HeroldKelly T. HughesDavid A. HunstadAlexander H\u00fcttenhoferMichael IbbaAkiko IwasakiJorgen JohanssonAndreas KapplerKrystyna M. KazmierczakCorby KistlerDavid M. KnipeLaura J. KnollArash KomeiliRichard J. KuhnMamuka KvaratskheliaNathaniel Roy LandauStanley M. LemonKim LewisDouglas R. LowyRebecca Marie LynchAndrew J. MacphersonWilliam MargolinEric MartensRachel M. McLoughlinDennis W. MetzgerJoachim Morschh\u00e4userJoseph MougousChristian MunzKevin MylesJulie OverbaughAndreas PeschelEverett C. PesciThomas PietschmannAnn M. PowersRichard A. ProctorMatthew Mark RamseyTimothy D. ReadAnthony R. RichardsonGloria Marcela RodriguezForest RohwerAntonis RokasTony RomeoSusan M. RosenbergPhilip C. RosenstielGian Maria RossoliniMonica J. RothCraig R. RoyRuth M. RuprechtMichael J. SadowskyJohn-Demian SauerChrista M. SchleperViviana SimonAbraham L. SonensheinJustin SonnenburgManjula SritharanGurol SuelNobuhiro SuzukiDaniel J. ThornhillRaphael ValdiviaSusana T. ValenteMarjan W. van der WoudeStefanie N. VogelMark J. WalkerJens WalterDavid S. WeissRoy D. WelchAngus C. WilsonAlan J. WolfeOtto O. YangFitnat H. YildizVincent B. YoungJae-Hyuk YuGongyi ZhangXinning ZhangThe time and effort of the following experts in handling articles have been essential to ensuring the high quality of our publications, and their help is greatly appreciated."} +{"text": "Present: The 4th author's name, \u2018Ding-wei Wu,\u2019 is spelled incorrectly.Correct: The proper spelling is \u2018Wei-ding Wu\u2019.https://doi.org/10.18632/oncotarget.15513Original article: Oncotarget. 2017; 8:23155-23166."} +{"text": "Present: The current funding information is incomplete.Correct: The complete funding information is given below.https://doi.org/10.18632/oncotarget.13056Original article: Oncotarget. 2017; 8:15943-15951."} +{"text": "Present: The original figure Correct: The proper spelling appears in the figure below. The authors sincerely apologize for this error.10.18632/oncotarget.9251Original article: Oncotarget. 2016; 7:41217-41232. doi:"} +{"text": "Baylisascaris procyonis\u2013Associated Meningoencephalitis in a Previously Healthy Adult, California, USA . The article has been corrected online (http://wwwnc.cdc.gov/eid/article/22/8/15-1939_article).The name of author Natalie Witek was misspelled in"} +{"text": "Present: The acknowledgement information is incomplete.Correct: Additional acknowledgements are shown below.ACKNOWLEDGEMENTSThis work was co-financed by European Development Regional Fund (ERDF).10604-16. doi: 10.18632/oncotarget.3550Original article: Oncotarget. 2015; 6:"} +{"text": "This article has been corrected: The correct Figure https://doi.org/10.18632/oncotarget.23046Original article: Oncotarget. 2017; 8:115164-115178."} +{"text": "Present: Due to a technical error during image processing, the same picture set of control cells were used in TEM figures for both HCT116 wt and p53\u2212/\u2212 cells. Figure Corrected: Correct Figure 43679-97. doi: 10.18632/oncotarget.6030.Original article: Oncotarget. 2015; 6(41):"} +{"text": "R. Soc. open sci.4, 170543. (Published 20 December 2017). (doi:10.1098/rsos.170543)The first sentence of \u00a74.2 currently reads \u2018Our pilot study (which was pre-registered with Open Science Framework: - osf.io),\u2026\u2019.https://osf.io/qbp7h/),\u2026\u2019.It should read: \u2018Our pilot study (which was pre-registered with Open Science Framework:"} +{"text": "There are errors in the Author Contributions. The correct contributions are: Conceptualization: LDP FARZ JDC. Data curation: LDP. Formal analysis: LDP FARZ. Funding acquisition: FARZ JDC MCGR. Investigation: LDP FARZ JDC MO ALCM ERJ APRT. Methodology: LDP FARZ JDC. Project administration: FARZ JDC MCGR. Resources: FARZ JDC MCGR. Software: LDP FARZ. Writing \u2013 original draft: LDP FARZ. Writing \u2013 review & editing: LDP FARZ JDC MO ALCM ERJ APRT."} +{"text": "Lactic acid is shown to derive from sugar derived from corn or sugar beets.https://www.natureworksllc.comNatureWorks is a company making polylactic acid from lactic acid that has been produced by a microbial fermentation process.https://link.springer.com/chapter/10.1007/978-3-662-45209-7_74 platform chemistry.Succinic acid is emerging as an important bio\u2010product, due to its value in the food industry and also for the\u00a0chemical industry as a potential replacement for the current maleic anhydride Chttps://www1.eere.energy.gov/bioenergy/pdfs/ibr_arra_myriant.pdfThis page describes a biorefinery for producing bio\u2010succinic acid, located in the Port of Lake Providence, Louisiana, USA. The production capacity is 30 million pounds annually.https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3572532/Aspergillus species.This paper deals with metabolic reactions producing itaconic acid. Major microbial produces are https://www.omicsonline.org/open-access/fermentative-itaconic-acid-production-2376-0214.1000119.php?aid=28089This review covers itaconic acid, a platform chemical that is important in textile and pharmaceutical industries.https://en.wikipedia.org/wiki/Malolactic_fermentationThe malolactic acid fermentation is most important in the wine industry. During the process, the tart\u2010flavoured malic acid is transformed into the softer\u2010tasting product, lactic acid.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89131999000300001Aspergillus niger and Candida sp.This review article is on citric acid production. Citric acid is the largest organic acid fermentation product, and it is produced mainly by https://ccj.springeropen.com/articles/10.1186/s13065-017-0251-yThis review article brings out the important point that citric acid is produced in 4 billion ton quantities by biomass fermentation processes.https://genome.jgi.doe.gov/Aspni5/Aspni5.home.htmlAspergillus niger.This is the genome portal page for a major citric acid producing microorganism, https://www.omicsonline.org/scholarly/acetic-acid-fermentation-journals-articles-ppts-list.phpThis page on fermentation technology is focused on acetic acid fermentation. Some relevant links are provided to articles and conference proceedings.http://www.mdpi.com/2311-5637/3/2/21Propionic acid is produced through microbial fermentation. It has applications in the manufacture of cosmetics and pharmaceuticals.https://www.sciencedirect.com/topics/agricultural-and-biological-sciences/alpha-linolenic-acidAlpha\u2010linoleic acid is considered an essential fatty acid for humans; and so, it is sought after by the food industry.http://www.sciencedirect.com/science/article/pii/S1369703X16302789Yarrowia lipolytica.Gamma\u2010linolenic acid is valued for pharmaceutical and nutraceutical applications. The present paper discusses its production using the yeast"} +{"text": "Biol. Lett.12, 20160586. (Published online 18 October 2016) (doi:10.1098/rsbl.2016.0586)The caption for"} +{"text": "Present: Due to an error in proofreading, figure Corrected: The proper figure 30831-30849. doi: 10.18632/oncotarget.5036.Original article: Oncotarget. 2015; 6(31):"} +{"text": "Phil. Trans. R. Soc. B372, 20160274 (Published Online 11 September 2017) (doi:10.1098/rstb.2016.0274)The name of the first author, Marc Ringelhan, was spelt incorrectly as \u2018Marc Ringehan\u2019. Therefore, the author list should appear as follows:Marc Ringelhan, Jane A. McKeating and Ulrike Protzer"} +{"text": "Nucl. Acids Res. 42 (12): 7489\u20137527. doi: 10.1093/nar/gku447The authors wish to correct reference 228 from:228. Naito T., Kobayashi I. Cell death programmed by selfish genes\u2013or, why are there restriction enzymes. Jikken Igaku 1995;13:1444\u20131447.To228. Naito T., Kusano K., Kobayashi I. Selfish behavior of restriction-modification systems. Science 1995;267:897\u2013899."} +{"text": "Present section: Due to an error made during submission of the revised files, acknowledgements were omitted from this manuscript.Correct section: Corrected acknowledgements information is provided below. Authors sincerely apologize for this oversight.10433-47. doi: 10.18632/oncotarget.7197.Original article: Oncotarget. 2016; 7(9):"} +{"text": "This article has been corrected: Dr. Shanbao Cai was moved to the first position in the author list.The authors sincerely apologize for this oversight.28420-28434. https://doi.org/10.18632/oncotarget.8546Original article: Oncotarget. 2016; 7:"} +{"text": "Nature Communications8: Article number: 15653; DOI: 10.1038/ncomms15653 (2017); Published: 06152017; Updated: 083020172-weighted phantom images in reverse order, and Supplementary Fig. 4d is a duplicate of Fig. 3d. The corrected version of Supplementary Fig. 4 is shown below as Supplementary Fig. 4 of this Article contains errors. Supplementary Fig. 4c shows the T"} +{"text": "Unfortunately both the HTML and the PDF versions of this article on the SChanges to in-text citations in HTML VersionDeletion of hyperlinks in HTML version to tables in other documentsCorrection of URLs in HTML versionhttp://leg2.state.va.us/DLS/h&sdocs.nsf/Search+All/?SearchView&SearchOrder=4&query=38.2-3419.1In Table 1, URL should be: http://leg2.state.va.us/DLS/h&sdocs.nsf/Search+All/?SearchView&earchOrder=4&query=38.2-3419.1In Footnote 4, URL should be: Changes to in-text citations in PDF VersionCorrection of URLs in PDF versionhttp://leg2.state.va.us/DLS/h&sdocs.nsf/Search+All/?SearchView&SearchOrder=4&query=38.2-3419.1In Table 1, URL should be: http://leg2.state.va.us/DLS/h&sdocs.nsf/Search+All/?SearchView&SearchOrder=4&query=38.2-3419.1In Endnote 2, URL should be: http://www.heritage.org/research/testimony/2013/06/health-care-consolidation-and-competition-after-ppaca. Accessed on September 13, 2016.In Endnote 11, URL should be:"} +{"text": "Scientific Reports 10.1038/s41598-018-21892-y, published online 27 February 2018Correction to: In this Article, the Data availability section was omitted from the Additional Information section:http://www.ncbi.nlm.nih.gov/bioproject/396973.Data availability: The raw sequences archive has been deposited to GenBank: BioProject ID PRJNA396973"} +{"text": "In the original publication the nameIncorrect version:Jeremy M. Aymar.Correct version:Jeremy M. Aymard."} +{"text": "Present: Due to an error in grant support.Correct: The updated grant support are given below. The author sincerely apologizes for this error.10.18632/oncotarget.7897.Original article: Oncotarget. 2016; 7(15):20902-18. doi:"} +{"text": "R. Soc. open sci.4, 160759 (Published online 22 March 2017). (doi:10.1098/rsos.160759)1.b and the resulting Rankine reciprocal is incorrect in fig. 9c. The correct arrangements are shown below.This correction concerns fig. 9 of"} +{"text": "This erratum corrects article: \u2033 Antithyroid drug induced agranulocytosis: what still we need to learn \u2033 The Pan African Medical Journal. 2016;23:27. doi:10.11604/pamj.2016.23.27.8365 The original version of this article containe"} +{"text": "Biol. Lett.11, 20150303. (Published online 8 July 2015) (doi:10.1098/rsbl.2015.0303)et al. [http://dx.doi.org/10.5061/dryad.pr318 [The data associated with Rutledge et al. are now ad.pr318 ."} +{"text": "Present: Due to an error made during the assembly of Figure Correct: Correct Figure 14179-90. doi: 10.18632/oncotarget.3651.Original article: Oncotarget. 2015; 6(16):"} +{"text": "Mycobacterium tuberculosis Infection among Asian Elephants in Captivity . The article has been corrected online (https://wwwnc.cdc.gov/eid/article/23/3/16-0726_article).The name of author Apurva Narechania was misspelled in"} +{"text": "Adv. Sci.2015, 2, 1400006Vtot vs. fluorosurfactant concentration was inadvertently missed. Figure The authors wish to correct Figure"} +{"text": "This article has an addendum: The below information has been added to this article. https://crln.acrl.org/index.php/crlnews/article/view/16837/18434https://doi.org/10.18632/oncotarget.24272Original article: Oncotarget. 2018; 9:5515-5516."} +{"text": "The correct reference is: Posavac HD, Posavac SS, Posavac EJ. Exposure to media images of female attractiveness and concern with body weight among young women. Sex Roles. 1998; 38: 187\u2013201. doi:10.1002/eat.20663.Reference 15 is incorrect. The correct reference is: Glauert R, Rhodes G, Fink B, Grammer K. Body dissatisfaction and attentional bias to thin bodies. Int J Eat Disord. 2009; 43: 42\u201349. doi:10.1002/eat.20703.Reference 19 is incorrect. The correct reference is: Smith E, Rieger E. An investigation of the effect of body dissatisfaction on selective attention toward negative shape and weight-related information. Int J Eat Disord. 2010; 43: 358\u2013364. doi:10.1023/A:1023910315561.Refence 27 is incorrect. The correct reference is: Treynor W. Rumination reconsidered: A psychometric analysis. Cognit Ther Res. 2003; 27: 247\u2013259. doi:There is an error in"} +{"text": "Nature Communications6: Article number: 8029; DOI: 10.1038/ncomms9029 (2015); Published 08202015; Updated 02072017Professors SonBinh T. Nguyen and Jiaxing Huang were inadvertently omitted from the list of corresponding authors in this Article. The correct information for correspondence is: \u2018Correspondence and requests for materials should be addressed to H.D.E. espinosa@northwestern.edu) or to J.H. jiaxing-huang@northwestern.edu) or to S.T.N. stn@northwestern.edu)."} +{"text": "Approximately 24 million U.S. homes contain deteriorated lead-based paint and lead-contaminated house dust (https://www.cdc.gov/nceh/lead/.Additional information about childhood lead poisoning prevention is available at"} +{"text": "Open Biol. 6, 160240. (Published online 7 December 2016). (doi:10.1098/rsob.160240)A correction is required for the manuscript detailed below: in Table 2, the significance levels for variable (16) should read: \u22120.010 and \u22120.460**."} +{"text": "Assessing the relationship between lung cancer and metabolic conditions is challenging because of the confounding effect of tobacco. Mendelian randomization (MR), or the use of genetic instrumental variables to assess causality, may help to identify the metabolic drivers of lung cancer.2]), but not for adenocarcinoma (OR [95%CI] = 0.93 [0.79\u20131.08]) (Pheterogeneity= 4.3x10-3). Additional analysis using a genetic instrument for BMI showed that each SD increase in BMI increased cigarette consumption by 1.27 cigarettes per day (P = 2.1x10-3), providing novel evidence that a genetic susceptibility to obesity influences smoking patterns. There was also evidence that low-density lipoprotein cholesterol was inversely associated with lung cancer overall risk (OR [95%CI] = 0.90 [0.84\u20130.97] per SD of 38 mg/dl), while fasting insulin was positively associated (OR [95%CI] = 1.63 [1.25\u20132.13] per SD of 44.4 pmol/l). Sensitivity analyses including a weighted-median approach and MR-Egger test did not detect other pleiotropic effects biasing the main results.We identified genetic instruments for potential metabolic risk factors and evaluated these in relation to risk using 29,266 lung cancer cases and 56,450 controls. The MR risk analysis suggested a causal effect of body mass index (BMI) on lung cancer risk for two of the three major histological subtypes, with evidence of a risk increase for squamous cell carcinoma (odds ratio (OR) [95% confidence interval (CI)] = 1.20 [1.01\u20131.43] and for small cell lung cancer (OR [95%CI] = 1.52 [1.15\u20132.00]) for each standard deviation (SD) increase in BMI [4.6 kg/mOur results are consistent with a causal role of fasting insulin and low-density lipoprotein cholesterol in lung cancer etiology, as well as for BMI in squamous cell and small cell carcinoma. The latter relation may be mediated by a previously unrecognized effect of obesity on smoking behavior. Lung cancer is the leading cause of cancer mortality . Most luMendelian randomization (MR) is an analytical approach based on genetic markers of an exposure that is less sensitive to reverse causation and confounding than traditional regression analyses in observational studies . InheritThe goal of the current study was to use genetic variations associated with a range of metabolic factors, including obesity, body shape, dyslipidemia and hyperglycemia, to further investigate the causal relationship between these metabolic exposures and lung cancer. The method applied in this study is called two-sample MR, which combines summary statistics on genetic variant-exposure and genetic variant-outcome associations from different samples ,21. Furt2 < 0.01 in European 1000 Genomes Phase3 samples = 0.93 [0.79\u20131.08]), but an increased risk for squamous cell carcinoma (OR [95%CI] = 1.20 [1.01\u20131.43]) and small cell lung cancer (OR [95%CI] = 1.52 [1.15\u20132.00]) (Pheterogeneity = 0.10) . Analyse = 0.10) . Weighte = 0.10) . The MR- = 0.10) , with MR = 0.10) . Funnel = 0.10) or histo = 0.10) . Sensiti = 0.10) Tables. There was no consistent evidence of a causal effect of HDL on lung cancer overall , nor anyPheterogeneity= 0.16) . Stratif= 0.16) , with a = 0.16) .Fasting glucose showed little evidence for an association with overall lung cancer risk , nor for-3; MR likelihood-based approach), while the other metabolic genetic instruments were not associated with cigarette smoking (MR likelihood-based estimate per SD increase [95%CI] = -0.35 [-0.92:0.22] for LDL and 1.21 [-0.67:3.08] for fasting insulin). Additionally, each SD increase in LDL was inversely associated with BMI , while each SD increase in fasting was not associated with BMI ).Using MR-Base, we also evaluated the association between cigarette smoking for each genetic instrument associated with lung cancer risk, including BMI, LDL, and fasting insulin. Each SD increase in BMI increased cigarette consumption by 1.27 cigarettes Click here for additional data file.S2 Fig2: between-strata heterogeneity. PHet: P value of between-strata heterogeneity.95%CI: 95% Confidence Interval; P: P value. I(PDF)Click here for additional data file.S3 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S4 Fig2: between-strata heterogeneity. PHet: P value of between-strata heterogeneity.95%CI: 95% Confidence Interval; P: P value. I(PDF)Click here for additional data file.S5 Fig2: between-strata heterogeneity. PHet: P value of between-strata heterogeneity.95%CI: 95% Confidence Interval; P: P value. I(PDF)Click here for additional data file.S6 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S7 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S8 Fig2: between-strata heterogeneity. PHet: P value of between-strata heterogeneity.95%CI: 95% Confidence Interval; P: P value. I(PDF)Click here for additional data file.S9 Fig2: between-strata heterogeneity. PHet: P value of between-strata heterogeneity.95%CI: 95% Confidence Interval; P: P value. I(PDF)Click here for additional data file.S10 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S11 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S12 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S13 Fig2: between-strata heterogeneity. PHet: P value of between-strata heterogeneity.95%CI: 95% Confidence Interval; P: P value. I(PDF)Click here for additional data file.S14 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S15 Fig2: between-strata heterogeneity. PHet: P value of between-strata heterogeneity.95%CI: 95% Confidence Interval; P: P value. I(PDF)Click here for additional data file.S16 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S17 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S18 Fig2: between-strata heterogeneity. PHet: P value of between-strata heterogeneity.95%CI: 95% Confidence Interval; P: P value. I(PDF)Click here for additional data file.S19 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S20 Fig2: between-strata heterogeneity. PHet: P value of between-strata heterogeneity.95%CI: 95% Confidence Interval; P: P value. I(PDF)Click here for additional data file.S21 FigOR: Odds ratio; Int: Intercept; P: P value.(PDF)Click here for additional data file.S1 TableCHR: Chromosome. BP: Base pair. SE: Standard error. BMI: Body mass index. HDL: High-density lipoprotein, LDL: Low-density lipoprotein.(PDF)Click here for additional data file.S2 TableHDL: High-density lipoprotein, LDL: Low-density lipoprotein. Chol: Cholesterol. OR: Odds ratio. LCI: Lower confidence interval. UCI: Upper confidence interval. P: P value.(PDF)Click here for additional data file.S3 TableInt: Intercept. HDL: High-density lipoprotein, LDL: Low-density lipoprotein. Chol: Cholesterol. OR: Odds ratio. Est: Estimate. LCI: Lower confidence interval. UCI: Upper confidence interval. P: P value.(PDF)Click here for additional data file."} +{"text": "Scientific Reports 10.1038/s41598-017-04773-8, published online 10 July 2017Correction to: The original PDF version of this Article contained an error in the order of corresponding authors.Correspondence and requests for materials should be addressed to S.Y. (email: shufen_yang@126.com) or J.Z. (email: jin_zhouxy@163.com) or J.S. now reads:Correspondence and requests for materials should be addressed to J.S. or J.Z. (email: jin_zhouxy@163.com) or S.Y. (email: shufen_yang@126.com)This has now been corrected in the PDF version of this Article."} +{"text": "Present: The grant support section is incomplete.Correct: The complete grant support information appears below.4373-4386. https://doi.org/10.18632/oncotarget.13875Original article: Oncotarget. 2017; 8:FUNDINGFunds from Fondo Europeo de Desarrollo Regional (FEDER) and by grants from the Ministry of Economy and Competitiveness of Spain and Comunidad Aut\u00f3noma de Madrid grant S2010/BMD- 2470 (Oncocycle Program) to J.M. Paramio."} +{"text": "Present: The images of cell migration were mistakenly duplicated at Fig.Correct: The corrected images appear below. The authors apologize for any confusion this error may have caused.28540-55. doi: 10.18632/oncotarget.8677Original article: Oncotarget. 2016; 7:"} +{"text": "Present: Due to an error during figure assembly, Figure Correct: The proper Figure 10.18632/oncotarget.3397Original article: Oncotarget. 2015; 6:11434-46. doi:"} +{"text": "VOLUME 105105(3) July, page 242http://dx.doi.org/10.5195/jmla.2017.98.Kokol P. Trend analysis of journal metrics: a new academic library service? J Med Libr Assoc. 2017 Jul;105(3):240\u20132. DOI:"} +{"text": "DOI: 10.1093/sysbio/syx080.Garba M.K., Nye T.M.W., Boys R.J. 2017. Probabilistic Distances Between Trees. Sys. Biol. The following sentence was incorrect:dTV.\u201d\u201cThe total variation metric between distributions is defined by The correct sentence is below:\u201cThe total variation metric between distributions is defined by This has now been corrected in the original article."} +{"text": "Nature Communications7: Article number: 12976 doi: ; DOI: 10.1038/ncomms12976 (2016); Published: 11232016; Updated: 11232016Drosophila larva in Fig. 3a is reproduced from the website http://www.prokop.co.uk/Research/Drosi-Info/nerve-cords.html with the kind permission of Dr Andreas Prokop.In Fig. 3 of this Article, an image attribution was inadvertently omitted. The drawing of a"} +{"text": "Seasonal flu vaccine uptake in persons aged 65 years and olderEpi-Insight 2016;17(10)October 2016, Irelandhttp://ndsc.newsweaver.ie/epiinsight/pnpyh4q4puin0ykqgxo0gg?a=1&p=50883926&t=17517774Food- and waterborne diseasesSharp increase in campylobacter infectionFolkh\u00e4lsomyndigheten website3 October 2016, Swedenhttps://www.folkhalsomyndigheten.se/nyheter-och-press/nyhetsarkiv/2016/oktober/kraftig-okning-av-infektion-med-campylobacter/An oyster-associated norovirus outbreak following a corporate dinner \u2013 cohort study, Toulouse (France), January 2015Bulletin \u00e9pid\u00e9miologique hebdomadaire, 26\u2013276 September 2016, Francehttp://invs.santepubliquefrance.fr/beh/2016/26-27/pdf/2016_26-27.pdfHealthcare-associated infections and antibiotic resistanceCandidaemia in England, Wales and Northern Ireland: 2015Surveillance of Health Protection Report; 10(32)23 September 2016, UKhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/555332/hpr3216_cnddm.pdfPseudomonas spp. and Stenotrophomonas spp. in England: 2008\u20132015Health Protection Report; 10(32)Voluntary surveillance of bacteraemia caused by 23 September 2016, UKhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/555333/hpr3216_psdmns.pdfImmunisation \u2013 measles, mumps rubella, whooping cough and vaccine uptakeHPS Weekly Report, 2016;50(40)4 October 2016, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2016/1640.pdfQuarterly vaccination coverage statistics for children aged up to five years in the UK (COVER programme): April to June 2016Health Protection Report; 10(33)30 September 2016, UKhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/556895/HPR_COVER_Q1617_1_April_to_June_2016_v4.pdfLaboratory confirmed reports of invasive meningococcal disease in England: April to June 2016Health Protection Report; 10(33)30 September 2016, UKhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/557329/hpr3316_IMD.pdfPreliminary vaccine coverage estimates for the meningococcal B (MenB) immunisation programme for England, update to the end of August 2016Health Protection Report; 10(32)23 September 2016, UKhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/555057/hpr3216_menB.pdfNational rotavirus immunisation programme update: preliminary vaccine coverage for England, February 2016 to July 2016Health Protection Report; 10(32)23 September 2016, UKhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/555048/hpr3216_rtvrs_VC.pdfVaccine coverage estimate for the GP based catch-up meningococcal ACWY (MenACWY) immunisation programme for school leavers (becoming 18 before 31 August 2016) in England, cumulative data to end-August 2016Health Protection Report; 10(32)19 September 2016, UKhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/553736/hpr3216_menACWY_VCa.pdfMeningococcal disease in 2015EPI-NEWS 37, 201614 September 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2037%20-%202016.aspxMeasles cases in the Copenhagen areaEPI-NEWS 36, 20167 September 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2036%20-%202016.aspxSexually transmitted diseasesSyphilis in Scotland 2015: updateHPS Weekly Report, 2016;50(39)27 September 2016, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2016/1639.pdfChlamydia trachomatis and Neisseria gonorrhoeae infection in Scotland: laboratory diagnoses 2006 \u2013 2015HPS Weekly Report, 2016;50(39)27 September 2016, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2016/1639.pdfGonococcal antibiotic surveillance in Scotland (GASS): prevalence, patterns and trends in 2015HPS Weekly Report, 2016;50(39)27 September 2016, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2016/1639.pdfGonorrhoea 2015EPI-NEWS 38, 201621 September 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2038%20-%202016.aspxSyphilis 2015EPI-NEWS 36, 20167 September 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2036%20-%202016.aspxZoonoses and vector-borne diseasesIncidence of protozoal infection reported to HPS in 2015HPS Weekly Report, 2016;50(37)13 September 2016, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2016/1637.pdfGeneral bacterial and protozoal outbreaks of infectious intestinal disease reported to HPS in 2015HPS Weekly Report, 2016;50(36)6 September 2016, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2016/1636.pdfEscherichia coli (STEC) in the Netherlands, 2015Surveillance of Shiga-toxin producing Infectieziekten Bulletin 2016; 27(7)September 2016, the Netherlandshttp://www.rivm.nl/dsresource?objectid=rivmp:321901&type=org&disposition=inline&ns_nc=1HepatitidesHepatitis E outbreak associated with the consumption of a spit-roasted piglet, Brittany (France), 2013Bulletin \u00e9pid\u00e9miologique hebdomadaire, 26\u2013276 September 2016, Francehttp://invs.santepubliquefrance.fr/beh/2016/26-27/pdf/2016_26-27.pdfHepatitis B vaccination for injecting drug users: from vaccination programme to individual careInfectieziekten Bulletin 2016; 27(7)September 2016, the Netherlandshttp://www.rivm.nl/dsresource?objectid=rivmp:321901&type=org&disposition=inline&ns_nc=1OtherPseudomonas infections associated with ear-piercing after-care productsHealth Protection Report; 10(31)16 September 2016, UKhttps://www.gov.uk/government/publications/health-protection-report-volume-10-2016/hpr-volume-10-issue-31-news-16-september"} +{"text": "Present: Two images of the same scene with different brightness were accidentally included in Figure Correct: The proper Figure https://doi.org/10.18632/oncotarget.10017Original article: Oncotarget. 2016; 7:46042-46055."} +{"text": "In 2011, life expectancy at birth was 78.7 years for the total U.S. population, 76.3 years for males, and 81.1 years for females. Life expectancy was highest for Hispanics for both males and females. In each racial/ethnic group, females had higher life expectancies than males. Life expectancy ranged from 71.7 years for non-Hispanic black males to 83.7 years for Hispanic females.Source: National Center for Health Statistics. Deaths: final data for 2011. Available at http://www.cdc.gov/nchs/data/nvsr/nvsr63/nvsr63_03.pdf.Reported by: Arialdi Minino, aminino@cdc.gov, 301-458-4376."} +{"text": "References 1, 2 and 3 are incomplete. The correct references should read as follows:1. Forsburg SL, Nurse P. Cell Cycle Regulation in the Yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. Annual Review of Cell Biology. 1991; 7(1):227\u201356. PMID: 18093482. Murray AW. Recycling the Cell Cycle: Cyclins Revisited. Cell. 2004; 116(2):221\u201334. PMID: 1474443310.1016/j.cub.2004.11.027 PMID: 155891393. Jorgensen P, Tyers M. How Cells Coordinate Growth and Division. Current Biology. 2004; 14(23):R1014-27. doi:The publisher apologizes for this error."} +{"text": "This article has been corrected: The correct author name is given below:Juan R. Vi\u00f1a9100-9113. https://doi.org/10.18632/oncotarget.23888Original article: Oncotarget. 2018; 9:"} +{"text": "Present: Due to an error made during the assembly of Figure Correct: Correct Figure 12326-39. doi: 10.18632/oncotarget.3619.Original article: Oncotarget. 2015; 6(14):"} +{"text": "This article has been corrected: the institution information was updated to include the postal code.The authors sincerely apologize for this oversight.23360-23375. https://doi.org/10.18632/oncotarget.15579Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: The institutional affiliation information for Dr. Gennaro Ciliberto was updated to include affiliation #17.The authors sincerely apologize for this oversight.https://doi.org/10.18632/oncotarget.16725Original article: Oncotarget. 2017; 8:30606-30616."} +{"text": "Present: Due to an error in the production process, figures Corrected: The proper figure legends are provided below. The publisher sincerely apologizes for this oversight.20410-24. doi: 10.18632/oncotarget.7804.Original article: Oncotarget. 2016; 7(15):"} +{"text": "This article has been corrected: the 1st affiliation has been revised.https://doi.org/10.18632/oncotarget.21365Original article: Oncotarget. 2017; 8:84417-84425."} +{"text": "Healthcare-associated infections and antibiotic resistanceLaboratory surveillance of candidaemia in England, Wales and Northern Ireland: 2016Health Protection Report; 11(32)15 September 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/645312/hpr3217_cnddmia2016.pdfStaphylococcus aureus bacteraemia in England, Wales and Northern Ireland: 2016Laboratory surveillance of Health Protection Report; 11(29)18 August 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/638891/hpr2917_stph-aurs.pdfPseudomonas spp. and Stenotrophomonas spp. bacteraemia in England, Wales and Northern Ireland: 2016Laboratory surveillance of Health Protection Report; 11(25)21 July 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/631374/hpr2517_psdmns.pdfFood- and waterborne diseasesSalmonella Kottbus eventsEpidemiological and microbiological investigation of temporally parallel food-related Epidemiologisches Bulletin 38, 201721 September 2017, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2017/Ausgaben/38_17.pdf?__blob=publicationFileHepatitis A outbreak linked to the contamination of products from a bakery, Herault (France), 2014Bulletin \u00e9pid\u00e9miologique hebdomadaire, 2119 September 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/21/pdf/2017_21.pdfIncidence of protozoal infection reported to HPS in 2016HPS Weekly Report 2017; 51(37)19 September 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1737.pdfSalmonella in the Netherlands in 2016. In humans, farm animals and in foodTrends in Infectieziekten Bulletin 2017; 28(7)14 September 2017, the Netherlandshttp://www.rivm.nl/Documenten_en_publicaties/Algemeen_Actueel/Uitgaven/Infectieziekten_Bulletin/Jaargang_28_2017/Infectieziekten_Bulletin_jaargang_28_nummer_7_september_2017/Inhoud_september_2017/Trends_in_Salmonella_in_Nederland_in_2016_Bij_de_mens_landbouwhuisdieren_en_in_voedselEscherichia coli (STEC) in the Netherlands, 2016Surveillance of shigatoxin-producing Infectieziekten Bulletin 2017; 28(7)14 September 2017, the Netherlandshttp://www.rivm.nl/Documenten_en_publicaties/Algemeen_Actueel/Uitgaven/Infectieziekten_Bulletin/Jaargang_28_2017/Infectieziekten_Bulletin_jaargang_28_nummer_7_september_2017/Inhoud_september_2017/Surveillance_van_shigatoxine_producerende_Escherichia_coli_STEC_in_Nederland_2016STEC in Scotland 2016: enhanced surveillance and reference laboratory dataHPS Weekly Report 2017; 51(32)15 August 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1732.pdfHepatitidesAcute hepatitis B (England): annual report for 2016Health Protection Report; 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11(30)25 August 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/640460/hpr3017_mmr.pdfIncrease of invasive infections with group A streptococci in January\u2013April 2017EPI-NEWS 27a+b, 20175 July 2017, Denmarkhttps://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2027a%20-%202017.aspxInvasive meningococcal disease (laboratory reports in England): January to March 2017Health Protection Report; 11(23)30 June 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/624164/hpr2317_IMD.pdfLaboratory confirmed cases of Pertussis (England): January to March 2017Health Protection Report; 11(23)30 June 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/624162/hpr2317_prtsss.pdfQuarterly vaccination coverage statistics for children aged up to five years in the UK (COVER programme): January to March 2017Health Protection Report; 11(23)30 June 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/623860/hpr2317_COVER.pdfPneumococcal Polysaccharide Vaccine (PPV) coverage report, England, April 2016 to March 2017Health Protection Report; 11(23)30 June 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/624133/hpr2317_PPV.pdfRespiratory diseasesLegionellosis in Scotland 2015\u20132016HPS Weekly Report 2017; 51(34)29 August 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1734.pdfSexually transmitted diseasesHIV infection and AIDS: Quarterly report to 30 June 2017HPS Weekly Report 2017; 51(36)12 September 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1736.pdfHIV 2016EPI-NEWS 36, 20176 September 2017, Denmarkhttps://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2036%20-%202017.aspxScreening of pregnant women for hepatitis B, HIV and syphilis, 2016EPI-NEWS 35, 201730 August 2017, Denmarkhttps://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2035%20-%202017.aspxChlamydia 2016EPI-NEWS 34, 201723 August 2017, Denmarkhttps://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2034%20-%202017.aspxSyphilis in Scotland 2016: updateHPS Weekly Report 2017; 51(33)22 August 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1733.pdfSexually transmitted infections: updating data of two sentinel surveillance systems active in Italy on 31 December 2015Not Ist Super Sanit\u00e0 2017;30(7-8)July\u2212August 2017, Italyhttp://www.iss.it/binary/publ/cont/ONLINE_lug_ago_2017.pdfSentinel surveillance of sexually transmitted infections based on clinical microbiology laboratoriesNot Ist Super Sanit\u00e0 2017;30(7-8)July\u2212August 2017, Italyhttp://www.iss.it/binary/publ/cont/ONLINE_lug_ago_2017.pdfAnnual report from the sentinel surveillance study of blood borne virus testing in England: data for January to December 2016Health Protection Report; 11(26)28 July 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/633797/hpr2617_bbv-ss.pdfHIV prevalence estimate among men who have sex with men attending gay venues in five French cities \u2013 PREVAGAY 2015Bulletin \u00e9pid\u00e9miologique hebdomadaire, 1818 July 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/18/pdf/2017_18.pdfOutbreak of hepatitis A among men who have sex with men, Rouen (France), December 2016 - April 2017Bulletin \u00e9pid\u00e9miologique hebdomadaire, 1818 July 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/18/pdf/2017_18.pdfZoonoses and vector-borne diseases Common animal associated infections quarterly report : second quarter 2017Health Protection Report; 11(28)11 August 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/637401/hpr2817_zoos.pdfTravel health: Malaria reported in Scotland 2016HPS Weekly Report 2017; 51(28)18 July 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1728.pdfOtherInfectious disease surveillance of migrant populations in Calais and Grande-Synthe, France, November 2015 \u2013 October 2016Bulletin \u00e9pid\u00e9miologique hebdomadaire, 19-205 September 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/19-20/pdf/2017_19-20.pdfCreutzfeldt-Jakob disease (CJD) biannual update (August 2017)Health Protection Report; 11(29)18 August 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/640136/hpr2917_CJD.pdfSeroprevalence of HIV infections, syphilis, hepatitis B, hepatitis C and hepatitis A in asylum seekers in SaxonyEpidemiologisches Bulletin 29, 201720 July 2017, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2017/Ausgaben/29_17.pdf?__blob=publicationFile"} +{"text": "Carex rostrata var. borealis and C. stenolepis, Two Problematic Taxa in Carex Section Vesicariae (Cyperaceae). PLoS ONE 11(10): e0165430. doi:10.1371/journal.pone.0165430.The first author\u2019s surname is incorrectly preceded by an initial. The correct citation is: Pedersen ATM, Nowak MD, Brysting AK, Elven R, Bjor\u00e5 CS (2016) Hybrid Origins of"} +{"text": "Colistin resistance in Gram-negative bacteria - the situation in GermanyEpidemiologisches Bulletin 46, 201621 November 2016, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2016/Ausgaben/46_16.pdf?__blob=publicationFileOnline survey on influenza vaccination among hospital staffEpidemiologisches Bulletin 47, 201628 November 2016, Germanyhttp://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2016/Ausgaben/47_16.pdf?__blob=publicationFileStreptococcus pneumoniae causing bacteraemia in England, Wales and Northern Ireland: 2015Surveillance of Health Protection Report; 10(40)18 November 2016, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/571420/hpr4016.pdfEpidemic occurrence of whooping coughEPI-NEWS 46, 201616 November 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2046%20-%202016.aspxThe first year with statutory registration of vaccinations in the Danish Vaccination RegisterEPI-NEWS 45, 20169 November 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2045%20-%202016.aspxSpecial edition: World AIDS Day, December 1, 2016Bulletin \u00e9pid\u00e9miologique hebdomadaire, 41-4229 November 2016, Francehttp://invs.santepubliquefrance.fr/beh/2016/41-42/pdf/2016_41-42.pdfHIV infection and AIDS: Quarterly report to 30 September 2016HPS Weekly Report, 2016;50(48)28 November 2016, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2016/1648.pdfEstimated number of new HIV infections and estimated total number of people with HIV in GermanyEpidemiologisches Bulletin 45, 201614 November 2016, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2016/Ausgaben/45_16.pdf?__blob=publicationFileSelected parasite infections detected using the PCR methodEPI-NEWS 44, 20162 November 2016, Denmarkhttp://www.ssi.dk/English/News/EPI-NEWS/2016/No%2044%20-%202016.aspxSpecial edition: Asylum seekers and infectious diseasesInfectieziekten Bulletin 2016; 27(9)November 2016, the Netherlandshttp://www.rivm.nl/Documenten_en_publicaties/Algemeen_Actueel/Uitgaven/Infectieziekten_Bulletin/Jaargang_27_2016/November_2016/Inhoud_november_2016"} +{"text": "Present: Due to an error during manuscript preparation, loading control (GAPDH) was omitted from Figure Corrected: Correct Figure 38166-80. doi: 10.18632/oncotarget.6241.Original article: Oncotarget. 2015; 6(35):"} +{"text": "This article has been corrected: the authors first and last names order was updated.74910-74916. https://doi.org/10.18632/oncotarget.20456Original article: Oncotarget. 2017; 8:"} +{"text": "The fourth author\u2019s name is spelled incorrectly. The correct name is: Joanna L. Elson.https://doi.org/10.1371/journal.pone.0186802The corrected citation is: Tomas C, Brown A, Strassheim V, Elson JL, Newton J, Manning P (2017) Cellular bioenergetics is impaired in patients with chronic fatigue syndrome. PLoS ONE 12(10): e0186802."} +{"text": "Biochemia Medica 2017;27(2):279-84. DOI: https://doi.org/10.11613/BM.2017.030. This is a correction for This Erratum corrects the section Potential conflict of interest, which was originally published. The publisher apologizes for this error."} +{"text": "During 2009\u20132011, higher death rates for COPD among persons aged \u226555 years were associated with more rural localities, with rates increasing steadily from the least to the most rural county. For males, the age-adjusted COPD death rate in rural counties was 59% higher than in large central metropolitan counties . For females, the age-adjusted COPD death rate in rural counties was 39% higher than in large central metropolitan counties . COPD death rates for males were 21% to 47% higher than for females, with the largest differentials observed in nonmetropolitan counties .Sources: National Vital Statistics System. County-level mortality file. Available at http://www.cdc.gov/nchs/deaths.htm and http://wonder.cdc.gov/mortsql.html.http://www.cdc.gov/nchs/data/series/sr_02/sr02_166.pdf.Ingram DD, Franco SJ. 2013 NCHS urban-rural classification scheme for counties. Vital Health Stat 2014;2(166). Available at Reported by: Deborah D. Ingram, PhD, ddingram@cdc.gov, 301-458-4733."} +{"text": "Present: Due to an error made during the final figure assembly, blots for U266 were mistakenly used in lieu of blots for JJN3 in Figure Correct: Corrected Figure 9296-308. doi: 10.18632/oncotarget.6974.Original article: Oncotarget. 2016; 7(8):"} +{"text": "An error in the author byline has been corrected. The new citation is: Baluch F, Itti L (2010) Training Top-Down Attention Improves Performance on a Triple-Conjunction Search Task. PLoS ONE 5(2): e9127. doi:10.1371/journal.pone.0009127."} +{"text": "The first author's name is spelled incorrectly. The correct name is Alexei V. Samsonovich. The citation should read: Samsonovich AV, Ascoli GA (2010) Principal Semantic Components of Language and the Measurement of Meaning. PLoS ONE 5(6): e10921. doi:10.1371/journal.pone.0010921"} +{"text": "British Journal of Cancer (2002) 87, 1339\u20131339. doi:10.1038/sj.bjc.6600561www.bjcancer.com\u00a9 2002 Cancer Research UKCorrection to:British Journal of Cancer (2001) 84, 625\u2013929. doi:10.1054/sj.bjc.2001.1975 Unfortunately due to an error The correct version is printed below."} +{"text": "Correction for:Caenorhabditis elegans. PLoS Biol 5(10): e259. doi:10.1371/journal.pbio.0050259Rea SL, Ventura N, Johnson TE (2007) Relationship between mitochondrial electron transport chain dysfunction, development, and life extension in reas3@uthscsa.edu.The corresponding author's e-mail is incorrect. It should be:"} +{"text": "Michal Lapidot's name was incorrectly listed as Lapidot Michal. The correct citation is: Lapidot M, Mizrahi-Man O, Pilpel Y (2008) Functional Characterization of Variations on Regulatory Motifs. PLoS Genet 4(3): e1000018. doi:10.1371/journal.pgen.1000018"} +{"text": "Correction to:British Journal of Cancer (2005) 92, 2225\u20132232. doi:10.1038/sj.bjc.6602632Owing to an author error,"} +{"text": "Correction to:British Journal of Cancer (2001) 85, 1211\u20131218. doi:10.1054/bjoc.2001.2053"} +{"text": "British Journal of Cancer (2002) 87, 1340\u20131340. doi:10.1038/sj.bjc.6600571www.bjcancer.com\u00a9 2002 Cancer Research UKCorrection to:British Journal of Cancer (2002) 86, 1479\u20131486. doi:10.1038/sj.bjc.6600297 In the above paper an error occurred in the title.The correct title is printed below:c release in breast cancer cellsThe bisphosphonate zoledronic acid impairs Ras membrane localisation and induces cytochrome"} +{"text": "Correction to:British Journal of Cancer (1999) 80, 1359\u20131365. doi:10.1038/sj.bjc.6690529"} +{"text": "In the following article, references were misnumbered. Reference number 4 should be treated as reference number 3, and vice-versa.ViewpointKaur M., Develop advocacy for public health 2008; 33(2):71-72Correct references should be as follows:http://www.healthpromotionjournal.com/publications/global/1999-09/1999-09.htm3. Weinstein Y, Kaluski N. Country profile: Nutrition and education in Israel, Health Promotion: Global Perspectives, 1999 Vol. 2 No.4 Available from: 4. Chapman S. Public health advocacy: A primer. J Epidemiol Comm Health 2004;58:361-5"} +{"text": "The second author's name contained an error. Sim Xueling should be Xueling Sim. The correct citation is: Ikram MK, Sim X, Jensen RA, Cotch MF, Hewitt AW, et al. (2010) Four Novel Loci Influence the Microcirculation In Vivo. PLoS Genet 6(10): e1001184. doi:10.1371/journal.pgen.1001184."} +{"text": "British Journal of Cancer (2002) 86, 1981\u20131985. DOI: 10.1038/sj.bjc.6600444www.bjcancer.comCancer Research UK\u00a9 2002"} +{"text": "British Journal of Cancer (2002) 87, 1479\u20131479. doi:10.1038/sj.bjc.6600636www.bjcancer.com\u00a9 2002 Cancer Research UK Correction to:British Journal of Cancer (2002) 86, 917\u2013923. doi: 10.1038/sj.bjc.6600156We have reviewed our data about the cloning of the novel putative SIR-like gene and we accept Dr Frye's assertions . We are"} +{"text": "British Journal of Cancer (2002) 86, 1973\u20131980. DOI: 10.1038/sj.bjc.6600443www.bjcancer.comCancer Research UK\u00a9 2002"} +{"text": "The fourth author's name was spelled incorrectly. The correct name is: Ivan A. Adzhubei. The correct citation is: Schmidt S, Gerasimova A, Kondrashov FA, Adzhubei IA, Kondrashov AS, et al. (2008) Hypermutable Non-Synonymous Sites Are under Stronger Negative Selection. PLoS Genet 4(11): e1000281. doi:10.1371/journal.pgen.1000281"} +{"text": "Correction for:10.1371/journal.pbio.0050300Kelsch W, Mosley CP, Lin CW, Lois C (2007) Distinct mammalian precursors are committed to generate neurons with defined dendritic projection patterns. PLoS Biol 5(11): e300. doi:"} +{"text": "Our survey paper containIn , page 7,In , page 10http://www.diabetesnet.com/diabetes_technology/insulin_pump_models.php;[50] http://www.childrenwithdiabetes.com/continuous.htm.[51] In , page 15"} +{"text": "Correction to:British Journal of Cancer (2000) 83, 1664\u20131673. doi:10.1054/bjoc.2000.1501"} +{"text": "British Journal of Cancer (2002) 87, 1480\u20131480. doi:10.1038/sj.bjc.6600652www.bjcancer.com\u00a9 2002 Cancer Research UKCorrection to:British Journal of Cancer 2001; 85 (Suppl 2): 1\u20135. doi:10.1054/bjoc.2001.1981 An error has been noted within Figure 1"} +{"text": "British Journal of Cancer (2002) 86, 658. DOI: 10.1038/sj/bjc/6600055www.bjcancer.comCancer Research UK\u00a9 2002 With a research interest in angiogenesis and antiangiogenesis , I have Recently,"} +{"text": "The first author's name is not spelled correctly. It should be: Dale J. Hedges. The correct citation is: Hedges DJ, Burges D, Powell E, Almonte C, Huang J, et al. (2009) Exome Sequencing of a Multigenerational Human Pedigree. PLoS ONE 4(12): e8232. doi:10.1371/journal.pone.0008232"} +{"text": "The fourth author's name appears incorrectly. It should be: Hong Y. Choi. The correct citation should read: Ohazama A, Johnson EB, Ota MS, Choi HY, Porntaveetus T, et al. (2008) Lrp4 Modulates Extracellular Integration of Cell Signaling Pathways in Development. PLoS ONE 3(12): e4092. doi:10.1371/journal.pone.0004092"} +{"text": "This URL is missing a tilde. The correct URL reads: http://www.biostat.wisc.edu/~aasmith/catcode/"} +{"text": "British Journal of Cancer (2002) 86, 1666\u20131666. DOI: 10.1038/sj/bjc/6600308www.bjcancer.comCancer Research UK\u00a9 2002 Correction to:British Journal of Cancer (2001) 85, 493-496. DOI: 10.1054/bjoc.2001.1979The authors would like to acknowledge the Research into Childhood Cancer charity (RICC) for funding for the above project."} +{"text": "Salmonella typhi. Ann Saudi Med [serial online] 2010 [cited 2010 Sep 5];30:313-6. Available from:http://www.saudiannals.net/text.asp?2010/30/4/313/65267Somily AM. An imported enteric fever caused by a quinolone-resistant Salmonella terminology was applied. The term Salmonella Typhi (and abbreviated form S Typhi) should have been used in the title and elsewhere, according to current nomenclature .Incorrect"} +{"text": "Ageing: a global perspective. Volume 12, Issue 29, 1999.Keeffe J. Vision assessment and prescription of low vision devices. Comm Eye Health J 2004;17(49): 3\u20134.Minto H and Awan H. Establishing low vision services at secondary level. Comm Eye Health J 2004;17(49): 5.Watkinson S. Visual impairment in older people: the nurse's role. Nursing Standard 2005;19(17): 45\u201352. (Available to dowload free of charge from www.nursing-standard.co.uk)McNaughton et al. Low Vision Assessment. Butterworth-Heinemann Ltd, 2000. UK \u00a320 (plus postage and packing). Available from Waterstones: 71\u201374 North Street, Brighton, East Sussex BN1 1ZA, UK.manager@brighton.waterstones.co.ukEmail: Helpage International: www.helpage.orgLow Vision Online: www.lowvisiononline.unimelb.edu.auWHO: ageing and life course: www.who.int/ageing/en/Low Vision Kit. Includes information, E-charts and various materials to test vision and learn more about low vision. US $30 (incl. delivery). Available from the Centre for Eye Research Australia, 32 Gisborne Street, East Melbourne 3002, Victoria, Australia. Email: lowvisiononline-info@unimelb.edu.au"} +{"text": "The author contributions are incorrect. The correct author contributions should read as follows: Conceived and designed the experiments: XY RGS QY. Performed the experiments: XY MA, FS. Analyzed the data: XY RKMK PT. Contributed reagents/materials/analysis tools: RKMK FS KY LDM PT. Wrote the paper: XY QY."} +{"text": "Although the Concise Guide represents approximately 400 pages, the material presented is substantially reduced compared to information and links presented on the website. It provides a permanent, citable, point\u2010in\u2010time record that will survive database updates. The full contents of this section can be found at http://onlinelibrary.wiley.com/doi/10.1111/bph.14748. G protein\u2010coupled receptors are one of the six major pharmacological targets into which the Guide is divided, with the others being: ion channels, nuclear hormone receptors, catalytic receptors, enzymes and transporters. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading. The landscape format of the Concise Guide is designed to facilitate comparison of related targets from material contemporary to mid\u20102019, and supersedes data presented in the 2017/18, 2015/16 and 2013/14 Concise Guides and previous Guides to Receptors and Channels. It is produced in close conjunction with the International Union of Basic and Clinical Pharmacology Committee on Receptor Nomenclature and Drug Classification (NC\u2010IUPHAR), therefore, providing official IUPHAR classification and nomenclature for human drug targets, where appropriate.The Concise Guide to PHARMACOLOGY 2019/20 is the fourth in this series of biennial publications. The Concise Guide provides concise overviews of the key properties of nearly 1800 human drug targets with an emphasis on selective pharmacology (where available), plus links to the open access knowledgebase source of drug targets and their ligands ( The remaining 350 non\u2010sensory GPCRs mediate signalling by ligands that range in size from small molecules to peptides to large proteins; they are the targets for the majority of drugs in clinical usage , although only a minority of these receptors are exploited therapeutically. The first classification scheme to be proposed for GPCRs [http://www.ncbi.nlm.nih.gov/pubmed/8081729?dopt=AbstractPlus] divided them, on the basic of sequence homology, into six classes. These classes and their prototype members were as follows: Class A (rhodopsin\u2010like), Class B (secretin receptor family), Class C (metabotropic glutamate), Class D , Class E (cyclic AMP receptors) and Class F (frizzled/smoothened). Of these, classes D and E are not found in vertebrates. An alternative classification scheme \"GRAFS\" [http://www.ncbi.nlm.nih.gov/pubmed/15862553?dopt=AbstractPlus] divides vertebrate GPCRs into five classes, overlapping with the A\u2010F nomenclature, viz:G protein\u2010coupled receptors (GPCRs) are the largest class of membrane proteins in the human genome. The term \"7TM receptor\" is commonly used interchangeably with \"GPCR\", although there are some receptors with seven transmembrane domains that do not signal through G proteins. GPCRs share a common architecture, each consisting of a single polypeptide with an extracellular N\u2010terminus, an intracellular C\u2010terminus and seven hydrophobic transmembrane domains (TM1\u2010TM7) linked by three extracellular loops (ECL1\u2010ECL3) and three intracellular loops (ICL1\u2010ICL3). About 800 GPCRs have been identified in man, of which about half have sensory functions, mediating olfaction (\u02dc400), taste (33), light perception (10) and pheromone signalling (5) .Rhodopsin family (http://www.guidetopharmacology.org/GRAC/GPCRListForward?class=A), which includes receptors for a wide variety of small molecules, neurotransmitters, peptides and hormones, together with olfactory receptors, visual pigments, taste type 2 receptors and five pheromone receptors (V1 receptors).http://www.guidetopharmacology.org/GRAC/GPCRListForward?class=Adhesion GPCRs are phylogenetically related to class B receptors, from which they differ by possessing large extracellular N\u2010termini that are autoproteolytically cleaved from their 7TM domains at a conserved \"GPCR proteolysis site\" (GPS) which lies within a much larger (320 residue) \"GPCR autoproteolysis\u2010inducing\" (GAIN) domain, an evolutionary ancient mofif also found in polycystic kidney disease 1 (PKD1)\u2010like proteins, which has been suggested to be both required and sufficient for autoproteolysis [http://www.ncbi.nlm.nih.gov/pubmed/23850273?dopt=AbstractPlus].http://www.guidetopharmacology.org/GRAC/GPCRListForward?class=Frizzled consists of 10 Frizzled proteins (FZD(1\u201010)) and Smoothened (SMO). The FZDs are activated by secreted lipoglycoproteins of the WNT family, whereas SMO is indirectly activated by the Hedgehog (HH) family of proteins acting on the transmembrane protein Patched (PTCH).Secretin family, encoded by 15 genes in humans. The ligands for receptors in this family are polypeptide hormones of 27\u2010141 amino acid residues; nine of the mammalian receptors respond to ligands that are structurally related to one another , glucose\u2010dependent insulinotropic polypeptide (GIP), secretin, vasoactive intestinal peptide (VIP), pituitary adenylate cyclase\u2010activating polypeptide (PACAP) and growth\u2010hormone\u2010releasing hormone (GHRH)) [http://www.ncbi.nlm.nih.gov/pubmed/11790261?dopt=AbstractPlus].aNumbers in brackets refer to orphan receptors for which an endogenous ligand has been proposed in at least one publication, see [http://www.ncbi.nlm.nih.gov/pubmed/23686350?dopt=AbstractPlus]; b[http://www.ncbi.nlm.nih.gov/pubmed/19129093?dopt=AbstractPlus]; c[http://www.ncbi.nlm.nih.gov/pubmed/15034552?dopt=AbstractPlus]; d[http://www.ncbi.nlm.nih.gov/pubmed/15774036?dopt=AbstractPlus].2 adrenoceptor, the latter culminating in the award of the 2012 Nobel Prize in chemistry to Robert Lefkowitz and Brian Kobilka .Much of our current understanding of the structure and function of GPCRs is the result of pioneering work on the visual pigment rhodopsin and on the \u03b2Below is a curated list of pseudogenes that in humans are non\u2010coding for receptor protein. In some cases these have a shared ancestry with genes that encode functional receptors in rats and mice.https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:19240, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:16341, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:4529, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:16291, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:7959, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:4513, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:31924, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:31925, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:19103, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:19106, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:19107, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:20615, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:20640, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:20641, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:20642, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:43898, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:43905, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:20616. A more detailed listing containg further information can be viewed http://www.guidetopharmacology.org/GRAC//DATA/GPCR_pseudogenes.xlsx.https://www.genenames.org/cgi\u2010bin/genefamilies/set/141, and curated by https://genome.weizmann.ac.il/horde/ and https://senselab.med.yale.edu/ordb/.Olfactory receptors are also seven\u2010transmembrane spanning G protein\u2010coupled receptors, responsible for the detection of odorants. These are not currently included as they are not yet associated with extensive pharmacological data but are curated in the following databases: The gene list of olfactory receptors at Mol. Pharmacol.93: 266\u2010269 [https://www.ncbi.nlm.nih.gov/pubmed/29348268?dopt=AbstractPlus]Kenakin T. (2018) Is the Quest for Signaling Bias Worth the Effort? et al. (2018) Biased Agonism in Drug Discovery\u2010Is It Too Soon to Choose a Path? Mol. Pharmacol.93: 259\u2010265 [https://www.ncbi.nlm.nih.gov/pubmed/29326242?dopt=AbstractPlus]Michel MC et al. (2017) Discovery of new GPCR ligands to illuminate new biology. Nat. Chem. Biol.13: 1143\u20101151 [https://www.ncbi.nlm.nih.gov/pubmed/29045379?dopt=AbstractPlus]Roth BL et al. (2018) G Protein\u2010Coupled Receptors as Targets for Approved Drugs: How Many Targets and How Many Drugs? Mol. Pharmacol.93: 251\u2010258 [https://www.ncbi.nlm.nih.gov/pubmed/29298813?dopt=AbstractPlus]Sriram K http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=706\u2013 http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=975\u2013This set contains \u2019orphan\u2019 G protein coupled receptors where the endogenous ligand(s) is not known, and other 7TM receptors.NC\u2010IUPHAR[http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus], for which preliminary evidence for an endogenous ligand has been published, or for which there exists a potential link to a disease, or disorder. These GPCRs have recently been reviewed in detail [http://www.ncbi.nlm.nih.gov/pubmed/23686350?dopt=AbstractPlus]. The GPCRs in Table 1 are all Class A, rhodopsin\u2010like GPCRs. Class A orphan GPCRs not listed in Table 1 are putative GPCRs with as\u2010yet unidentified endogenous ligands.Table 1 lists a number of putative GPCRs identified by Class A orphan GPCRs with putative endogenous ligandshttp://www.guidetopharmacology.org/GRAC/ObjectDisplayForward?objectId=89, http://www.guidetopharmacology.org/GRAC/ObjectDisplayForward?objectId=109 and http://www.guidetopharmacology.org/GRAC/ObjectDisplayForward?objectId=126 which are reported to respond to endogenous agents analogous to the endogenous cannabinoid ligands have been grouped together (http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=114).In addition the orphan receptors et al. (2015) Identification of a mast\u2010cell\u2010specific receptor crucial for pseudo\u2010allergic drug reactions. Nature519: 237\u201341 [https://www.ncbi.nlm.nih.gov/pubmed/25517090?dopt=AbstractPlus]McNeil BD This set contains class C \u2019orphan\u2019 G protein coupled receptors where the endogenous ligand(s) is not known.et al. (2017) Structural insight to mutation effects uncover a common allosteric site in class C GPCRs. Bioinformatics33: 1116\u20131120 [https://www.ncbi.nlm.nih.gov/pubmed/28011766?dopt=AbstractPlus]Harpse K http://www.ncbi.nlm.nih.gov/pubmed/12581520?dopt=AbstractPlus], TRPM5 [http://www.ncbi.nlm.nih.gov/pubmed/12581520?dopt=AbstractPlus] and IP3 [http://www.ncbi.nlm.nih.gov/pubmed/17925404?dopt=AbstractPlus] receptors in postreceptor signalling of taste receptors. Although predominantly associated with the oral cavity, taste receptors are also located elsewhere, including further down the gastrointestinal system, in the lungs and in the brain.Whilst the taste of acid and salty foods appear to be sensed by regulation of ion channel activity, bitter, sweet and umami tastes are sensed by specialised GPCR. Two classes of taste GPCR have been identified, T1R and T2R, which are similar in sequence and structure to Class C and Class A GPCR, respectively. Activation of taste receptors appears to involve gustducin\u2010 (G\u03b1t3) and G\u03b114\u2010mediated signalling, although the precise mechanisms remain obscure. Gene disruption studies suggest the involvement of PLC\u03b22 . T1R2/T1R3 heterodimers respond to sugars, such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5411, and artificial sweeteners, such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5432 [http://www.ncbi.nlm.nih.gov/pubmed/11509186?dopt=AbstractPlus].T1R3 acts as an obligate partner in T1R1/T1R3 and T1R2/T1R3 heterodimers, which sense umami or sweet, respectively. T1R1/T1R3 heterodimers respond to Positive allosteric modulators of T1R2/T1R3 have been reported [2363]. Such compounds enhance the sweet taste of sucrose mediated by these receptors, but are tasteless on their own.Pharmacol. Rev.71: 20\u201348 [https://www.ncbi.nlm.nih.gov/pubmed/30559245?dopt=AbstractPlus]Palmer RK. (2019) A Pharmacological Perspective on the Study of Taste. G protein\u2010coupled receptors \u2192 Orphan and other 7TM receptors \u2192 Taste 2 receptorshttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5433, but not 10 other bitter compounds [http://www.ncbi.nlm.nih.gov/pubmed/10761935?dopt=AbstractPlus], while T2R14 responded to at least eight different bitter tastants, including (\u2010)\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5344\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5344 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2291 [http://www.ncbi.nlm.nih.gov/pubmed/15178431?dopt=AbstractPlus].The composition and stoichiometry of bitter taste receptors is not yet established. Bitter receptors appear to separate into two groups, with very restricted ligand specificity or much broader responsiveness. For example, T2R5 responded to http://bitterdb.agri.huji.ac.il/dbbitter.php contains additional information on bitter compounds and receptors [http://www.ncbi.nlm.nih.gov/pubmed/21940398?dopt=AbstractPlus].Specialist database Pharmacol. Rev.71: 20\u201348 [https://www.ncbi.nlm.nih.gov/pubmed/30559245?dopt=AbstractPlus]Palmer RK. (2019) A Pharmacological Perspective on the Study of Taste. et al. (2013) International Union of Basic and Clinical Pharmacology. LXXXVIII. G protein\u2010coupled receptor list: recommendations for new pairings with cognate ligands. Pharmacol. Rev.65: 967\u201386 [https://www.ncbi.nlm.nih.gov/pubmed/23686350?dopt=AbstractPlus]Davenport AP et al. (2016) Insight into SUCNR1 (GPR91) structure and function. Pharmacol. Ther.159: 56\u201365 [https://www.ncbi.nlm.nih.gov/pubmed/26808164?dopt=AbstractPlus]Gilissen J et al. (2015) G Protein\u2010Coupled Receptor (GPCR) Expression in Native Cells: \u201cNovel\u201d endoGPCRs as Physiologic Regulators and Therapeutic Targets. Mol. Pharmacol.88: 181\u20107 [https://www.ncbi.nlm.nih.gov/pubmed/25737495?dopt=AbstractPlus]Insel PA et al. (2017) Neuro\u2010psychopharmacological perspective of Orphan receptors of Rhodopsin (class A) family of G protein\u2010coupled receptors. Psychopharmacology (Berl.)234: 1181\u20101207 [https://www.ncbi.nlm.nih.gov/pubmed/28289782?dopt=AbstractPlus]Khan MZ et al. (2017) The emerging pharmacology and function of GPR35 in the nervous system. Neuropharmacology113: 661\u2013671 [https://www.ncbi.nlm.nih.gov/pubmed/26232640?dopt=AbstractPlus]Mackenzie AE et al. (2016) Identifying ligands at orphan GPCRs: current status using structure\u2010based approaches. Br. J. Pharmacol.173: 2934\u201051 [https://www.ncbi.nlm.nih.gov/pubmed/26837045?dopt=AbstractPlus]Ngo T NC\u2010IUPHAR Subcommittee on 5\u2010HT receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/7938165?dopt=AbstractPlus] and subsequently revised [http://www.ncbi.nlm.nih.gov/pubmed/8936345?dopt=AbstractPlus]) are, with the exception of the ionotropic 5\u2010HT3 class, GPCRs where the endogenous agonist is http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5. The diversity of metabotropic 5\u2010HT receptors is increased by alternative splicing that produces isoforms of the 5\u2010HT2A , 5\u2010HT2C , 5\u2010HT4, 5\u2010HT6 and 5\u2010HT7 receptors. Unique amongst the GPCRs, RNA editing produces 5\u2010HT2C receptor isoforms that differ in function, such as efficiency and specificity of coupling to Gq/11 and also pharmacology . Most 5\u2010HT receptors (except 5\u2010ht1e and 5\u2010ht5b) play specific roles mediating functional responses in different tissues .5\u2010HT receptors displays a different pharmacology to the rodent forms of the receptor due to Thr335 of the human sequence being replaced by Asn in rodent receptors [http://www.ncbi.nlm.nih.gov/pubmed/18571247?dopt=AbstractPlus]. Wang et al. (2013) report X\u2010ray structures which reveal the binding modality of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=149 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=121 (DHE) to the 5\u2010HT1B receptor in comparison with the structure of the 5HT2B receptor [http://www.ncbi.nlm.nih.gov/pubmed/23519210?dopt=AbstractPlus]; some of these drugs adopt rather different conformations depending on the target receptor [http://www.ncbi.nlm.nih.gov/pubmed/29398112?dopt=AbstractPlus]. Various 5\u2010HT receptors have multiple partners in addition to G proteins, which may affect function and pharmacology [http://www.ncbi.nlm.nih.gov/pubmed/21777185?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3226 is a selective antagonist of the rodent 5\u2010HT1B receptor. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5434 (LSD) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=88 bind with high affinity to dopamine D4 and histamine H1 receptors respectively, and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=88 is a potent \u03b11 adrenoceptor antagonist, in addition to blocking 5\u2010HT2A receptors. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=139 (LSD) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=149 show a strong preference for arrestin recruitment over G protein coupling at the 5\u2010HT2B receptor, with no such preference evident at 5\u2010HT1B receptors, and they also antagonise 5\u2010HT7A receptors [http://www.ncbi.nlm.nih.gov/pubmed/23519215?dopt=AbstractPlus]. DHE (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=282), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=48 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=37 also show significant preference for arrestin recruitment over G protein coupling at 5\u2010HT2B receptors [http://www.ncbi.nlm.nih.gov/pubmed/23519215?dopt=AbstractPlus]. The 5\u2010HT2B (and other 5\u2010HT) receptors interact with immunocompetent cells [http://www.ncbi.nlm.nih.gov/pubmed/28265714?dopt=AbstractPlus]. The serotonin antagonist http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=206 was key to the discovery of the 5HT2C receptor [http://www.ncbi.nlm.nih.gov/pubmed/6519175?dopt=AbstractPlus], initially known as 5\u2010HT1C [http://www.ncbi.nlm.nih.gov/pubmed/28806488?dopt=AbstractPlus]. The human 5\u2010HT5A receptor may couple to several signal transduction pathways when stably expressed in C6 glioma cells [http://www.ncbi.nlm.nih.gov/pubmed/12558985?dopt=AbstractPlus] and rodent prefrontal cortex [http://www.ncbi.nlm.nih.gov/pubmed/22539842?dopt=AbstractPlus]. The human orthologue of the mouse 5\u2010ht5b receptor is non\u2010functional (stop codons); the 5\u2010ht1e receptor has not been cloned from mouse, or rat, impeding definition of its function [http://www.ncbi.nlm.nih.gov/pubmed/18571247?dopt=AbstractPlus]. In addition to accepted receptors, an \u2019orphan\u2019 receptor, unofficially termed5\u2010HT1P, has been described [http://www.ncbi.nlm.nih.gov/pubmed/10429737?dopt=AbstractPlus].Tabulated et al. (2011) 5\u2010HT(4) receptors, a place in the sun: act two. Curr Opin Pharmacol11: 87\u201093 [https://www.ncbi.nlm.nih.gov/pubmed/21342787?dopt=AbstractPlus]Bockaert J et al. (2011) 5\u2010HT receptors and reward\u2010related behaviour: a review. Neurosci Biobehav Rev35: 1419\u201049 [https://www.ncbi.nlm.nih.gov/pubmed/21402098?dopt=AbstractPlus]Hayes DJ et al. (1994) International Union of Pharmacology classification of receptors for 5hydroxytryptamine (Serotonin). Pharmacol. Rev.46: 157\u2010203 [https://www.ncbi.nlm.nih.gov/pubmed/7938165?dopt=AbstractPlus]Hoyer D et al. (2011)Serotonin 5\u2010HT7 receptor agents: Structure\u2010activity relationships and potential therapeutic applications in central nervous system disorders. Pharmacol. Ther.129: 120\u201048 [https://www.ncbi.nlm.nih.gov/pubmed/20923682?dopt=AbstractPlus]Leopoldo M et al. (2011) The role of serotonin receptors in the action of atypical antipsychotic drugs. Curr Opin Pharmacol11: 59\u201067 [https://www.ncbi.nlm.nih.gov/pubmed/21420906?dopt=AbstractPlus]Meltzer HY et al. (2012) The 5\u2010HT(7) receptor in learning and memory. Hippocampus22: 762\u201071 [https://www.ncbi.nlm.nih.gov/pubmed/21484935?dopt=AbstractPlus]Roberts AJ NC\u2010IUPHAR Subcommittee on Muscarinic Acetylcholine Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/9647869?dopt=AbstractPlus]) are GPCRs of the Class A, rhodopsin\u2010like family where the endogenous agonist is http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=294. In addition to the agents listed in the table, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=289, its structural analogues http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=334 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3271, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=333, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3258 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5435 have been described as functionally selective agonists of the M1 receptor subtype via binding in a mode distinct from that utilized by non\u2010selective agonists . There are two pharmacologically characterised allosteric sites on muscarinic receptors, one defined by it binding http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=356, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=347 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=342, and the other defined by the binding of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=337, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=340, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=339 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=346 .Muscarinic acetylcholine receptors and M4 receptors (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3262) . The allosteric site for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=356 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=347 on M2 receptors can be labelled by [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=364 [http://www.ncbi.nlm.nih.gov/pubmed/12815174?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=290 is a functionally selective partial agonist that appears to interact in a bitopic mode with both the orthosteric and an allosteric site on the M2 muscarinic receptor [http://www.ncbi.nlm.nih.gov/pubmed/18723515?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3255, hybrid 1 and hybrid 2, are multivalent (bitopic) ligands that also achieve selectivity for M2 receptors by binding both to the orthosteric and a nearby allosteric site .The crystal structures of the Me.g.http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=307, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=321, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=328) in order to identify the involvement of particular subtypes. It should be noted that the measured affinities of antagonists (and agonists) in radioligand binding studies are sensitive to ionic strength and can increase over 10\u2010fold at low ionic strength compared to their values at physiological ionic strengths [http://www.ncbi.nlm.nih.gov/pubmed/497538?dopt=AbstractPlus].Although numerous ligands for muscarinic acetylcholine receptors have been described, relatively few selective antagonists have been described, so it is common to assess the rank order of affinity of a number of antagonists of limited selectivity Toward an understanding of the structural basis of allostery in muscarinic acetylcholine receptors. J Gen Physiol150: 1360\u20101372 [https://www.ncbi.nlm.nih.gov/pubmed/30190312?dopt=AbstractPlus]Burger WAC et al. (1998) International Union of Pharmacology. XVII. Classification of muscarinic acetylcholine receptors. Pharmacol Rev50: 279\u2010290 [https://www.ncbi.nlm.nih.gov/pubmed/9647869?dopt=AbstractPlus]Caulfield MP Handb Exp Pharmacol 3\u201028 [https://www.ncbi.nlm.nih.gov/pubmed/22222692?dopt=AbstractPlus]Eglen RM. (2012) Overview of muscarinic receptor subtypes. et al. (2014) Muscarinic acetylcholine receptors: novel opportunities for drug development. Nat Rev Drug Discov13: 549\u201060 [https://www.ncbi.nlm.nih.gov/pubmed/24903776?dopt=AbstractPlus]Kruse AC et al. (2012) Structure\u2010function studies of muscarinic acetylcholine receptors. Handb Exp Pharmacol 29\u201048 [https://www.ncbi.nlm.nih.gov/pubmed/22222693?dopt=AbstractPlus]Leach K et al. (2012) The best of both worlds? Bitopic orthosteric/allosteric ligands of g proteincoupled receptors. Annu Rev Pharmacol Toxicol52: 153\u201078 [https://www.ncbi.nlm.nih.gov/pubmed/21910627?dopt=AbstractPlus]Valant C NC\u2010IUPHAR Subcommittee on Adenosine Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/11734617?dopt=AbstractPlus]) are activated by the endogenous ligand http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2844 . Crystal structures for the antagonist\u2010bound , agonist\u2010bound and G protein\u2010bound A2A adenosine receptors [http://www.ncbi.nlm.nih.gov/pubmed/27462812?dopt=AbstractPlus] have been described. The structures of an antagonist\u2010bound A1 receptor [http://www.ncbi.nlm.nih.gov/pubmed/28235198?dopt=AbstractPlus] and an adenosine\u2010bound A1 receptor\u2010Gi complex [http://www.ncbi.nlm.nih.gov/pubmed/29925945?dopt=AbstractPlus] have been resolved by cryo\u2010electronmicroscopy. Another structure of an antagonist\u2010bound A1 receptor obtained with X\u2010ray crystallography has also been reported [http://www.ncbi.nlm.nih.gov/pubmed/28712806?dopt=AbstractPlus].Adenosine receptors (2B adenosine receptor (https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:265) with 79% identity to the A2B adenosine receptor cDNA coding sequence, but which is unable to encode a functional receptor [http://www.ncbi.nlm.nih.gov/pubmed/7558011?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=386 also exhibits antagonism at A2B receptors . Antagonists at A3 receptors exhibit marked species differences, such that only http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=474 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=470 are selective at the rat A3 receptor. In the absence of other adenosine receptors, [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=406 and [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=455 can also be used to label A2B receptors (KD ca. 30 and 60 nM respectively). [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=436 also binds to A1 receptors [http://www.ncbi.nlm.nih.gov/pubmed/9459566?dopt=AbstractPlus]. . . http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3279 has been described as a fluorescent antagonist for labelling A1 adenosine receptors in living cells, although activity at other adenosine receptors was not examined [http://www.ncbi.nlm.nih.gov/pubmed/15070776?dopt=AbstractPlus].Adenosine inhibits many intracellular ATP\u2010utilising enzymes, including adenylyl cyclase (P\u2010site). A pseudogene exists for the Aet al. (2015) The A3 adenosine receptor: history and perspectives. Pharmacol Rev67: 74\u2010102 [https://www.ncbi.nlm.nih.gov/pubmed/25387804?dopt=AbstractPlus]Borea PA et al. (2017) Adenosine and adenosine receptors in the pathogenesis and treatment of rheumatic diseases. Nat Rev Rheumatol13: 41\u201051 [https://www.ncbi.nlm.nih.gov/pubmed/27829671?dopt=AbstractPlus]Cronstein BN et al. (2011) International Union of Basic and Clinical Pharmacology. LXXXI. Nomenclature and classification of adenosine receptors\u2013an update. Pharmacol Rev63: 1\u201034 [https://www.ncbi.nlm.nih.gov/pubmed/21303899?dopt=AbstractPlus]Fredholm BB et al. (2017) Kinetic Aspects of the Interaction between Ligand and G Protein\u2010Coupled Receptor: The Case of the Adenosine Receptors. Chem Rev117: 38\u201066 [https://www.ncbi.nlm.nih.gov/pubmed/27088232?dopt=AbstractPlus]Guo D et al. (2011) Allosteric modulation of adenosine receptors. Biochim Biophys Acta1808: 1309\u201018 [https://www.ncbi.nlm.nih.gov/pubmed/20599682?dopt=AbstractPlus]G\u00f6bly\u00f6s A Biochim Biophys Acta1808: 1284\u20109 [https://www.ncbi.nlm.nih.gov/pubmed/20888790?dopt=AbstractPlus]Lasley RD. (2011) Adenosine receptors and membrane microdomains. et al. (2011) Adenosine receptor desensitization and trafficking. Biochim Biophys Acta1808: 1319\u201028 [https://www.ncbi.nlm.nih.gov/pubmed/20550943?dopt=AbstractPlus]Mundell S et al. (2018) New paradigms in adenosine receptor pharmacology: allostery, oligomerization and biased agonism. Br J Pharmacol175: 4036\u20104046 [https://www.ncbi.nlm.nih.gov/pubmed/29679502?dopt=AbstractPlus]Vecchio EA etal. (2011)Normaland abnormalfunctions ofadenosine receptors inthecentral nervous system revealed by genetic knockout studies. Biochim Biophys Acta1808: 1358\u201079[https://www.ncbi.nlm.nih.gov/pubmed/21185258?dopt=AbstractPlus]Wei CJ http://www.ncbi.nlm.nih.gov/pubmed/22333914?dopt=AbstractPlus] containing a GPCR proteolytic site. The N\u2010terminus often shares structural homology with adhesive domains facilitating inter\u2010 and matricellular interactions and leading to the term adhesion GPCR . Several receptors have been suggested to function as mechanosensors . The nomenclature of these receptors was revised in 2015 as recommended byNC\u2010IUPHARand theAdhesion GPCR Consortium [http://www.ncbi.nlm.nih.gov/pubmed/25713288?dopt=AbstractPlus].Adhesion GPCRs are structurally identified on the basis of a large extracellular region, similar to the Class B GPCR, but which is linked to the 7TM region by a GPCR autoproteolysisinducing (GAIN) domain [Pharmacol Rev67: 338\u201067 [https://www.ncbi.nlm.nih.gov/pubmed/25713288?dopt=AbstractPlus]Hamann J et al. (2015) International Union of Basic and Clinical Pharmacology. XCIV. Adhesion G protein\u2010coupled receptors. Sci Signal6: re3 [https://www.ncbi.nlm.nih.gov/pubmed/23695165?dopt=AbstractPlus]Langenhan T et al. (2013) Sticky signaling\u2013adhesion class G protein\u2010coupled receptors take the stage. Handb Exp Pharmacol234: 111\u2010125 [https://www.ncbi.nlm.nih.gov/pubmed/27832486?dopt=AbstractPlus]Liebscher I et al. (2016) Tethered Agonism: A Common Activation Mechanism of Adhesion GPCRs. Mol Pharmacol88: 617\u201023 [https://www.ncbi.nlm.nih.gov/pubmed/25956432?dopt=AbstractPlus]Monk KR et al. (2015) Adhesion G Protein\u2010Coupled Receptors: From In Vitro Pharmacology to In Vivo Mechanisms. Annu Rev Pharmacol Toxicol58: 429\u2010449 [https://www.ncbi.nlm.nih.gov/pubmed/28968187?dopt=AbstractPlus]Purcell RH et al. (2018) Adhesion G Protein\u2010Coupled Receptors as Drug Targets. The nomenclature of the Adrenoceptors has been agreed by theNC\u2010IUPHAR Subcommittee on Adrenoceptors [http://www.ncbi.nlm.nih.gov/pubmed/7938162?dopt=AbstractPlus], see also [http://www.ncbi.nlm.nih.gov/pubmed/7568329?dopt=AbstractPlus].1\u2010Adrenoceptors are activated by the endogenous agonists (\u2010)\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=479 and (\u2010)\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=505. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=485, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=483 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=515 are agonists and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=503 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=515 antagonists considered selective for \u03b11\u2010 relative to \u03b12\u2010adrenoceptors. . Some tissues possess \u03b11A\u2010adrenoceptors that display relatively low affinity in functional and binding assays for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=503 indicative of different receptor states or locations. \u03b11A\u2010adrenoceptor C\u2010terminal splice variants form homo\u2010 and heterodimers, but fail to generate a functional \u03b11L\u2010adrenoceptor [http://www.ncbi.nlm.nih.gov/pubmed/15266013?dopt=AbstractPlus]. \u03b11D\u2010Adrenoceptors form heterodimers with \u03b11B\u2010 or \u03b22\u2010adrenoceptors that show increased cell\u2010surface expression [http://www.ncbi.nlm.nih.gov/pubmed/15615865?dopt=AbstractPlus]. Recombinant \u03b11D\u2010adrenoceptors have been shown in some heterologous systems to be mainly located intracellularly but cell\u2010surface localization is encouraged by truncation of the Nterminus, or by co\u2010expression of \u03b11B\u2010 or \u03b22\u2010adrenoceptors . In blood vessels all three \u03b11\u2010adrenoceptor subtypes are located on the surface and intracellularly . Signalling is predominantly via Gq/11 but\u03b11\u2010adrenoceptors also couple toGi/o, Gs and G12/13. Several \u03b11A\u2010adrenoceptor agonists display ligand directed signalling bias relative to noradrenaline [http://www.ncbi.nlm.nih.gov/pubmed/20978120?dopt=AbstractPlus]. There are also differences between subtypes in coupling efficiency to different pathways. In vascular smooth muscle, the potency of agonists is related to the predominant subtype, \u03b11D\u2010 conveying greater agonist sensitivity than \u03b11A\u2010adrenoceptors [http://www.ncbi.nlm.nih.gov/pubmed/23373597?dopt=AbstractPlus].The \u03b12\u2010Adrenoceptors are activated by (\u2010)\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=479 and with lower potency by (\u2010)\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=505. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=520 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5442 are agonists and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=136 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=102 antagonists selective for \u03b12\u2010 relative to \u03b11\u2010adrenoceptors. ; catecholamines have a low affinity, while rilmenidine and moxonidine are selective ligands evoking hypotensive effects in vivo. I1\u2010imidazoline receptors cause central inhibition of sympathetic tone, I2\u2010imidazoline receptors are an allosteric binding site on monoamine oxidase B, and I3\u2010imidazoline receptors regulate insulin secretion from pancreatic \u03b2\u2010cells. \u03b12A\u2010adrenoceptor stimulation reduces insulin secretion from \u03b2\u2010islets [http://www.ncbi.nlm.nih.gov/pubmed/22645144?dopt=AbstractPlus], with a polymorphism in the 5\u2019\u2010UTR of the ADRA2A gene being associated with increased receptor expression in \u03b2\u2010islets and heightened susceptibility to diabetes [http://www.ncbi.nlm.nih.gov/pubmed/19965390?dopt=AbstractPlus]. \u03b12A\u2010 and \u03b12C\u2010adrenoceptors form homodimers [http://www.ncbi.nlm.nih.gov/pubmed/16605244?dopt=AbstractPlus]. Heterodimers between \u03b12A\u2010 and either the \u03b12C\u2010adrenoceptor or \u03bc opioid peptide receptor exhibit altered signalling and trafficking properties compared to the individual receptors . Signalling by \u03b12\u2010adrenoceptors is primarily via Gi/o, although the \u03b12A\u2010adrenoceptor also couples to Gs [http://www.ncbi.nlm.nih.gov/pubmed/7559592?dopt=AbstractPlus]. Imidazoline compounds display bias relative to each other at the \u03b12A\u2010adrenoceptor [http://www.ncbi.nlm.nih.gov/pubmed/12649300?dopt=AbstractPlus]. The noradrenaline reuptake inhibitor desipramine acts directly on the \u03b12A\u2010adrenoceptor to promote internalisation via recruitment of arrestin [http://www.ncbi.nlm.nih.gov/pubmed/21859713?dopt=AbstractPlus].http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=479 and (\u2010)\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=505. Isoprenaline is selective for \u03b2\u2010adrenoceptors relative to \u03b11\u2010 and \u03b12\u2010adrenoceptors, while http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=564 (pKi 8.2\u20109.2) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=132 (pKi 10.011.0) are relatively \u03b21 and \u03b22 adrenoceptor\u2010selective antagonists. (\u2010)\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=505, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=538 and (\u2010)\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5571 show selectivity for \u03b21\u2010 relative to \u03b22\u2010adrenoceptors. Pharmacological differences exist between human and mouse \u03b23\u2010adrenoceptors, and the \u2019rodent selective\u2019 agonists http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=567 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3462 have low efficacy at the human \u03b23\u2010adrenoceptor whereas http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=532 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3931 activate human \u03b23\u2010adrenoceptors [88]. \u03b23\u2010Adrenoceptors are resistant to blockade by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=564, but can be blocked by high concentrations of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=550. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=547 has reasonably high affinity at \u03b23\u2010adrenoceptors, but does not discriminate well between the three \u03b2\u2010 subtypes whereas http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3931 is more selective. [125I]\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=132, [125I]\u2010hydroxy benzylpindolol and [3H]\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=563 are high affinity radioligands that label \u03b21\u2010 and \u03b22\u2010 adrenoceptors and \u03b23\u2010adrenoceptors can be labelled with higher concentrations (nM) of [125I]\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=132 together with \u03b21\u2010 and \u03b22\u2010adrenoceptor antagonists. [3H]\u2010L748337 is a \u03b23\u2010selective radioligand [http://www.ncbi.nlm.nih.gov/pubmed/24183974?dopt=AbstractPlus]. Fluorescent ligands such as BODIPY\u2010TMR\u2010CGP12177 can be used to track \u03b2adrenoceptors at the cellular level [8]. Somewhat selective \u03b21adrenoceptor agonists are used short term to treat cardiogenic shock but, chronically, reduce survival. \u03b21\u2010Adrenoceptor\u2010preferring antagonists areused totreathypertension , cardiac arrhythmias and cardiac failure . Cardiac failure is also treated with carvedilol that blocks \u03b21\u2010 and \u03b22\u2010adrenoceptors, as well as \u03b11\u2010adrenoceptors. Short and long acting \u03b22\u2010adrenoceptor\u2010selective agonists are powerful bronchodilators used to treat respiratory disorders. Many first generation \u03b2\u2010adrenoceptor antagonists (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=564) block both \u03b21\u2010 and \u03b22\u2010adrenoceptors and there are no \u03b22adrenoceptor\u2010selective antagonists used therapeutically. The \u03b23\u2010adrenoceptor agonist http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7445 is used to control overactive bladder syndrome.\u03b2\u2010Adrenoceptors are activated by the endogenous agonists (\u2010)\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=562 can be used to define \u03b21\u2010 or \u03b22adrenoceptors when conducted in the presence of a \u03b21\u2010 or \u03b22adrenoceptor\u2010selective antagonist. A fluorescent analogue of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=532 can be used to study \u03b22\u2010adrenoceptors in living cells [http://www.ncbi.nlm.nih.gov/pubmed/12770928?dopt=AbstractPlus]. , where the isoforms display different signalling characteristics [http://www.ncbi.nlm.nih.gov/pubmed/11959793?dopt=AbstractPlus]. There are 3 \u03b2\u2010adrenoceptors in turkey that have a pharmacology that differs from the human \u03b2\u2010adrenoceptors [http://www.ncbi.nlm.nih.gov/pubmed/21152092?dopt=AbstractPlus]. Numerous polymorphisms have been described for the \u03b2\u2010adrenoceptors; some are associated with signalling and trafficking, altered susceptibility to disease and/or altered responses to pharmacotherapy [http://www.ncbi.nlm.nih.gov/pubmed/15090197?dopt=AbstractPlus]. All \u03b2\u2010adrenoceptors couple to Gs (activating adenylyl cyclase and elevating cAMP levels), but also activate Gi and \u03b2\u2010arrestin\u2010mediated signalling. Many \u03b21\u2010 and \u03b22\u2010adrenoceptor antagonists are agonists at \u03b23adrenoceptors (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3462 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=569). Many \u2018antagonists\u2019 of cAMP accumulation, for example http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=551 and bucindolol, weakly activate MAP kinase pathways and thus display \u2019protean agonism\u2019. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=550 acts as a neutral antagonist in most systems so far examined. Agonists also display biased signalling at the \u03b22\u2010adrenoceptor via Gs or arrestins [http://www.ncbi.nlm.nih.gov/pubmed/18086673?dopt=AbstractPlus]. X\u2010ray crystal structures have been described of the agonist bound [http://www.ncbi.nlm.nih.gov/pubmed/21228877?dopt=AbstractPlus] and antagonist bound forms of the \u03b21[http://www.ncbi.nlm.nih.gov/pubmed/18594507?dopt=AbstractPlus], agonist\u2010bound [http://www.ncbi.nlm.nih.gov/pubmed/17962520?dopt=AbstractPlus] and antagonist\u2010bound forms of the \u03b22\u2010adrenoceptor , as well as a fully active agonistbound, Gs protein\u2010coupled \u03b22\u2010adrenoceptor [http://www.ncbi.nlm.nih.gov/pubmed/21772288?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=551 and bucindolol bind to a site on the \u03b21\u2010adrenoceptor involving contacts in TM2, 3, and 7 and extracellular loop 2 that may facilitate coupling to arrestins [http://www.ncbi.nlm.nih.gov/pubmed/18594507?dopt=AbstractPlus]. Compounds displaying arrestinbiased signalling at the \u03b22\u2010adrenoceptor have a greater effect on the conformation of TM7, whereas full agonists for Gs coupling promote movement of TM5 and TM6 [http://www.ncbi.nlm.nih.gov/pubmed/22267580?dopt=AbstractPlus]. Recent studies using NMR spectroscopy demonstrate significant conformational flexibility in the \u03b22\u2010adrenoceptor that is stabilized by both agonist and G proteins highlighting the dynamic nature of interactions with both ligand and downstreamsignalling partners . Such flexibility likely has consequences for our understanding of biased agonism, and for the future therapeutic exploitation of this phenomenon.[Trends Pharmacol. Sci.32: 227\u201034 [https://www.ncbi.nlm.nih.gov/pubmed/21429598?dopt=AbstractPlus]Baker JG et al. (2011) Evolution of \u03b2\u2010blockers: from anti\u2010anginal drugs to ligand\u2010directed signalling. Pharmacol. Rev.46: 121\u2010136 [https://www.ncbi.nlm.nih.gov/pubmed/7938162?dopt=AbstractPlus]Bylund DB et al. (1994) International Union of Pharmacology nomenclature of adrenoceptors. Br. J. Pharmacol.159: 1022\u201038 [https://www.ncbi.nlm.nih.gov/pubmed/20132209?dopt=AbstractPlus]Evans BA et al. (2010) Ligand\u2010directed signalling at beta\u2010adrenoceptors. J. Mol. Cell. Cardiol.51: 518\u201028 [https://www.ncbi.nlm.nih.gov/pubmed/21118696?dopt=AbstractPlus]Jensen BC et al. (2011) Alpha\u20101\u2010adrenergic receptors: targets for agonist drugs to treat heart failure. Trends Pharmacol. Sci.32: 213\u20108 [https://www.ncbi.nlm.nih.gov/pubmed/21414670?dopt=AbstractPlus]Kobilka BK. (2011) Structural insights into adrenergic receptor function and pharmacology. Trends Pharmacol. Sci.36: 196\u2010202 [https://www.ncbi.nlm.nih.gov/pubmed/25771972?dopt=AbstractPlus]Langer SZ. (2015) a2\u2010Adrenoceptors in the treatment of major neuropsychiatric disorders. Am. J. Physiol., Cell Physiol.308: C505\u201020 [https://www.ncbi.nlm.nih.gov/pubmed/25631871?dopt=AbstractPlus]Michel MC et al. (2015) Selectivity of pharmacological tools: implications for use in cell physiology. A review in the theme: Cell signaling: proteins, pathways and mechanisms. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2504 (Ang II) are mediated by AT1 and AT2 receptors , which have around 30% sequence similarity. The decapeptide http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=583, http://www.uniprot.org/uniprot/P01019), the octapeptide http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2504, http://www.uniprot.org/uniprot/P01019) and the heptapeptide http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=585, http://www.uniprot.org/uniprot/P01019) are endogenous ligands. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=590, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=587, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=592, etc. are clinically used AT1 receptor blockers.The actions of 1 receptors are predominantly coupled to Gq/11, however they are also linked to arrestin recruitment and stimulate G protein\u2010independent arrestin signalling [http://www.ncbi.nlm.nih.gov/pubmed/20427692?dopt=AbstractPlus]. Most species express a single AGTR1 gene, but two related agtr1a and agtr1b receptor genes are expressed in rodents. The AT2 receptor counteracts several of the growth responses initiated by the AT1 receptors. The AT2 receptor is much less abundant than the AT1 receptor in adult tissues and is upregulated in pathological conditions. AT1 receptor antagonists bearing substituted 4\u2010phenylquinoline moieties have been synthesized, which bind to AT1 receptors with nanomolar affinity and are slightly more potent than http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=590 in functional studies [http://www.ncbi.nlm.nih.gov/pubmed/15115399?dopt=AbstractPlus]. The antagonist activity of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3944 at the AT2 receptor has also been reported [http://www.ncbi.nlm.nih.gov/pubmed/3071214?dopt=AbstractPlus]. The AT1 and bradykinin B2 receptors have been proposed to form a heterodimeric complex [http://www.ncbi.nlm.nih.gov/pubmed/10993080?dopt=AbstractPlus]. \u03b2\u2010Arrestin1 prevents AT1\u2010B2 receptor heteromerization[http://www.ncbi.nlm.nih.gov/pubmed/30503206?dopt=AbstractPlus]. There is also evidence for an AT4 receptor that specifically binds http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5368 and is located in the brain and kidney. An additional putative endogenous ligand for the AT4 receptor has been described , a globin decapeptide) [http://www.ncbi.nlm.nih.gov/pubmed/9166749?dopt=AbstractPlus].ATet al. (2018) Crystal structure of the human angiotensin II type 2 receptor bound to an angiotensin II analog. Nat. Struct. Mol. Biol.25: 570\u2010576 [https://www.ncbi.nlm.nih.gov/pubmed/29967536?dopt=AbstractPlus]Asada H et al. (2015) International Union of Basic and Clinical Pharmacology. XCIX. Angiotensin Receptors: Interpreters of Pathophysiological Angiotensinergic Stimuli [corrected]. Pharmacol. Rev.67: 754\u2010819 [https://www.ncbi.nlm.nih.gov/pubmed/26315714?dopt=AbstractPlus]Karnik SS et al. (2019) Mechanism of Hormone Peptide Activation of a GPCR: Angiotensin II Activated State of AT_1R Initiated by van der Waals Attraction. J Chem Inf Model59: 373\u2010385 [https://www.ncbi.nlm.nih.gov/pubmed/30608150?dopt=AbstractPlus]Singh KD et al. (2019) Angiotensin Analogs with Divergent Bias Stabilize Distinct Receptor Conformations. Cell176: 468\u2010478.e11 [https://www.ncbi.nlm.nih.gov/pubmed/30639099?dopt=AbstractPlus]Wingler LM et al. (2019) Distinctive Activation Mechanism for Angiotensin Receptor Revealed by a Synthetic Nanobody. Cell176: 479\u2010490.e12 [https://www.ncbi.nlm.nih.gov/pubmed/30639100?dopt=AbstractPlus]Wingler LM et al. (2015) Structure of the Angiotensin receptor revealed by serial femtosecond crystallography. Cell161: 833\u201044 [https://www.ncbi.nlm.nih.gov/pubmed/25913193?dopt=AbstractPlus]Zhang H NC\u2010IUPHAR Subcommittee on the apelin receptornomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/20605969?dopt=AbstractPlus]) responds to apelin, a 36 amino\u2010acid peptide derived initially from bovine stomach. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=606 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=605, http://www.uniprot.org/uniprot/Q9ULZ1) and [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=599]http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=599 are the predominant endogenous ligands which are cleaved from a 77 amino\u2010acid precursor peptide (https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:16665http://www.uniprot.org/uniprot/Q9ULZ1) by a so far unidentified enzymatic pathway [http://www.ncbi.nlm.nih.gov/pubmed/9792798?dopt=AbstractPlus]. A second family of peptides discovered independently and named Elabela [http://www.ncbi.nlm.nih.gov/pubmed/24316148?dopt=AbstractPlus] or Toddler, that has little sequence similarity to apelin, is present, and functional at the apelin receptor in the adult cardiovascular system . Structure\u2010activity relationship Elabela analogues have been described [http://www.ncbi.nlm.nih.gov/pubmed/26986036?dopt=AbstractPlus].The apelin receptor . The apelin receptor may also act as a co\u2010receptor with CD4 for isolates of human immunodeficiency virus, with apelin blocking this function [http://www.ncbi.nlm.nih.gov/pubmed/11090199?dopt=AbstractPlus]. A modified apelin\u201013 peptide, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5354) was reported to block the hypotensive response to apelin in rat in vivo [http://www.ncbi.nlm.nih.gov/pubmed/15486224?dopt=AbstractPlus], however, this peptide exhibits agonist activity in HEK293 cells stably expressing the recombinant apelin receptor [http://www.ncbi.nlm.nih.gov/pubmed/12939143?dopt=AbstractPlus]. The apelin receptor antagonist, MM54, was reported to suppress tumour growth and increase survival in an intracranial xenograft mouse model of glioblastoma [http://www.ncbi.nlm.nih.gov/pubmed/29053791?dopt=AbstractPlus].Potency order determined for heterologously expressed human apelin receptor Neuroprotection of apelin and its signaling pathway. Peptides37: 171\u20103 [https://www.ncbi.nlm.nih.gov/pubmed/22820556?dopt=AbstractPlus]Cheng B et al. (2009) Structural insight into G\u2010protein coupled receptor binding by apelin. Biochemistry48: 537\u201048 [https://www.ncbi.nlm.nih.gov/pubmed/19123778?dopt=AbstractPlus]Langelaan DN et al. (2018) Vascular effects of apelin: Mechanisms and therapeutic potential. Pharmacol. Ther.190: 139\u2010147 [https://www.ncbi.nlm.nih.gov/pubmed/29807055?dopt=AbstractPlus]Mughal A et al. (2013) The apelin receptor APJ: journey from an orphan to a multifaceted regulator of homeostasis. J. Endocrinol.219: R13\u201035 [https://www.ncbi.nlm.nih.gov/pubmed/23943882?dopt=AbstractPlus]O\u2019Carroll AM et al. (2010) International Union of Basic and Clinical Pharmacology. LXXIV. Apelin receptor nomenclature, distribution, pharmacology, and function. Pharmacol. Rev.62: 331\u201042 [https://www.ncbi.nlm.nih.gov/pubmed/20605969?dopt=AbstractPlus]Pitkin SL et al. (2015) Apelin, Elabela/Toddler, and biased agonists as novel therapeutic agents in the cardiovascular system. Trends Pharmacol. Sci.36: 560\u20107 [https://www.ncbi.nlm.nih.gov/pubmed/26143239?dopt=AbstractPlus]Yang P http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2718. Selective agonists are promising drugs for the treatment of metabolic disorders, such as type II diabetes, obesity and atherosclerosis.The bile acid receptor (GPBA) responds to bile acids produced during the liver metabolism of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3945 has also been reported to inhibit inflammatory signalling through the NF\u03baB pathway [http://www.ncbi.nlm.nih.gov/pubmed/12960358?dopt=AbstractPlus]. Disruption of GPBA expression is reported to protect from cholesterol gallstone formation [http://www.ncbi.nlm.nih.gov/pubmed/16724960?dopt=AbstractPlus]. A new series of 5\u2010phenoxy\u20101,3\u2010dimethyl\u20101H\u2010pyrazole\u20104\u2010carboxamides have been reported as highly potent agonists [http://www.ncbi.nlm.nih.gov/pubmed/23337601?dopt=AbstractPlus].The triterpenoid natural product et al. (2014) The bile acid TGR5 membrane receptor: from basic research to clinical application. Dig Liver Dis46: 302\u201012 [https://www.ncbi.nlm.nih.gov/pubmed/24411485?dopt=AbstractPlus]Duboc H et al. (2014) GPBA: a GPCR for bile acids and an emerging therapeutic target for disorders of digestion and sensation. Br. J. Pharmacol.171: 1156\u201066 [https://www.ncbi.nlm.nih.gov/pubmed/24111923?dopt=AbstractPlus]Lieu T et al. (2009) Role of bile acids and bile acid receptors in metabolic regulation. Physiol. Rev.89: 147\u201091 [https://www.ncbi.nlm.nih.gov/pubmed/19126757?dopt=AbstractPlus]Lefebvre P et al. (2017) Clinical relevance of the bile acid receptor TGR5 in metabolism. Lancet Diabetes Endocrinol5: 224\u2010233 [https://www.ncbi.nlm.nih.gov/pubmed/27639537?dopt=AbstractPlus]van Nierop FS 1, BB2, BB3 . BB1 and BB2 are activated by the endogenous ligands http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=612 (GRP), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=613 (NMB) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3582) . http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=616 is a tetradecapeptide, originally derived from amphibians. The three Bn receptor subtypes couple primarily to the Gq/11 and G12/13 family of G proteins [http://www.ncbi.nlm.nih.gov/pubmed/18055507?dopt=AbstractPlus]. Each of these receptors is widely distributed in the CNS and peripheral tissues . Activation of BB1 and BB2 receptors causes a wide range of physiological/pathophysiogical actions, including the stimulation of normal and neoplastic tissue growth, smoothmuscle contraction, feeding behavior, secretion and many central nervous system effects including regulation of circadian rhythm and mediation of pruritus . A physiological role for the BB3 receptor has yet to be fully defined although recently studies suggest an important role in glucose and insulin regulation, metabolic homeostasis, feeding, regulation of body temperature, obesity, diabetes mellitus and growth of normal/neoplastic tissues .Mammalian bombesin (Bn) receptors comprise 3 subtypes: BBhttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=632,http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=632\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=632,http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=632,http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=632]http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=632) [http://www.ncbi.nlm.nih.gov/pubmed/9325344?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3946) has more than 200\u2010fold selectivity for BB3 receptors over BB1 and BB2 .All three human subtypes may be activated by [et al. (2015) Bombesin receptor subtype 3 as a potential target for obesity and diabetes. Expert Opin. Ther. Targets19: 1153\u201070 [https://www.ncbi.nlm.nih.gov/pubmed/26066663?dopt=AbstractPlus]Gonz\u00e1lez N et al. (2008) International Union of Pharmacology. LXVIII. Mammalian bombesin receptors: nomenclature, distribution, pharmacology, signaling, and functions in normal and disease states. Pharmacol. Rev.60: 1\u201042 [https://www.ncbi.nlm.nih.gov/pubmed/18055507?dopt=AbstractPlus]Jensen RT et al. (2017) Theranostic Prospects of Gastrin\u2010Releasing Peptide Receptor\u2010Radioantagonists in Oncology. PET Clin12: 297\u2010309 [https://www.ncbi.nlm.nih.gov/pubmed/28576168?dopt=AbstractPlus]Maina T et al. (2016) Bombesin related peptides/receptors and their promising therapeutic roles in cancer imaging, targeting and treatment. Expert Opin. Ther. Targets20: 1055\u201073 [https://www.ncbi.nlm.nih.gov/pubmed/26981612?dopt=AbstractPlus]Moreno P et al. (2018) Recent insights into biological functions of mammalian bombesin\u2010like peptides and their receptors. Curr Opin Endocrinol Diabetes Obes25: 36\u201041 [https://www.ncbi.nlm.nih.gov/pubmed/29120926?dopt=AbstractPlus]Qu X et al. (2015) Insights into bombesin receptors and ligands: Highlighting recent advances. Peptides72: 128\u201044 [https://www.ncbi.nlm.nih.gov/pubmed/25976083?dopt=AbstractPlus]Ramos\u2010\u00c1lvarez I NC\u2010IUPHAR subcommittee on Bradykinin (kinin) Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/15734727?dopt=AbstractPlus]) are activated by the endogenous peptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=649 (BK), [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=646]http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=646, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:6383), Lys\u2010BK ), [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=644]http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=644, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:6383), [Phospho\u2010Ser6]\u2010Bradykinin, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=639, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:6383) (Ile\u2010SerBK), [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3578]http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3578, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:6383) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3580]\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3580, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:6383). Variation in pharmacology and activity of B1 and B2 receptor antagonists at species orthologs has been documented. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=667 is approved in North America and Europe for the treatment of acute attacks of hereditary angioedema.Bradykinin (or kinin) receptors Non\u2010peptide antagonists for kinin B1 receptors: new insights into their therapeutic potential for the management of inflammation and pain. Trends Pharmacol. Sci.27: 646\u201051 [https://www.ncbi.nlm.nih.gov/pubmed/17056130?dopt=AbstractPlus]Campos MM et al. (2009) The kinin B(1) receptor and inflammation: new therapeutic target for cardiovascular disease. Curr Opin Pharmacol9: 125\u201031 [https://www.ncbi.nlm.nih.gov/pubmed/19124274?dopt=AbstractPlus]Duchene J et al. (2004) Bradykinin receptor ligands: therapeutic perspectives. Nat Rev Drug Discov3: 845\u201052 [https://www.ncbi.nlm.nih.gov/pubmed/15459675?dopt=AbstractPlus]Marceau F et al. (1999) Pharmacological characterization of the bradykinin B2 receptor: inter\u2010species variability and dissociation between binding and functional responses. Br. J. Pharmacol.126: 1083\u201090 [https://www.ncbi.nlm.nih.gov/pubmed/10204994?dopt=AbstractPlus]Paquet JL et al. (2010) Kinin receptor antagonists as potential neuroprotective agents in central nervous system injury. Molecules15: 6598\u2010618 [https://www.ncbi.nlm.nih.gov/pubmed/20877247?dopt=AbstractPlus]Thornton E et al. (2012) Discovery and therapeutic potential of kinin receptor antagonists. Expert Opin Drug Discov7: 1129\u201048 [https://www.ncbi.nlm.nih.gov/pubmed/23095011?dopt=AbstractPlus]Whalley ET NC\u2010IUPHAR Subcommittee on CGRP, AM, AMY, and CT receptorsnomenclature as agreed by the ) are generated by the genes https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:1440 (which codes for the CT receptor) and https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:16709 . Their function and pharmacology are altered in the presence of RAMPs (receptor activity\u2010modifying proteins), which are single TM domain proteins of ca. 130 amino acids, identified as a family of three members; RAMP1, RAMP2 and RAMP3. There are splice variants of the CT receptor; these in turn produce variants of the AMY receptor [http://www.ncbi.nlm.nih.gov/pubmed/12037140?dopt=AbstractPlus], some of which can be potently activated by CGRP. The endogenous agonists are the peptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=685 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=681\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=681, http://www.uniprot.org/uniprot/P06881) (formerly known as CGRP\u2010I), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=682 (formerly known as CGRP\u2010II), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=687 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=683 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=684 . There are species differences in peptide sequences, particularly for the CTs. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5369 {Pig} (CRSP) is another member of the family with selectivity for the CT receptor but it is not expressed in humans [http://www.ncbi.nlm.nih.gov/pubmed/12556539?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=702 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=703 are the most selective antagonists available, showing selectivity for CGRP receptors, with a particular preference for those of primate origin. CLR by itself binds no known endogenous ligand, but in the presence of RAMPs it gives receptors for CGRP, adrenomedullin and adrenomedullin 2/intermedin.This receptor family comprises a group of receptors for the calcitonin/CGRP family of peptides. The calcitonin (CT), amylin (AMY), calcitonin gene\u2010related peptide (CGRP) and adrenomedullin (AM) receptors has low affinity for 125I\u2010AMY binding sites, cells transfected with CTR and RAMPs can display potent CT functional responses. Transfection of human CTR with any RAMP can generate receptors with a high affinity for both salmon CT and AMY and varying affinity for different antagonists . The major human CTR splice variant (hCT(a), which does not contain an insert) with RAMP1 (i.e. the AMY1(a) receptor) has a high affinity for CGRP [http://www.ncbi.nlm.nih.gov/pubmed/26125036?dopt=AbstractPlus], unlike hCT(a)\u2010RAMP3 (i.e. AMY3(a) receptor) . However, the AMY receptor phenotype is RAMP\u2010type, splice variant and cell\u2010line\u2010dependent . Emerging data suggests that AMY1 could be a second CGRP receptor [http://www.ncbi.nlm.nih.gov/pubmed/29797087?dopt=AbstractPlus].It is important to note that a complication with the interpretation of pharmacological studies with AMY receptors in transfected cells is that most of this work has likely used a mixed population of receptors, encompassing RAMP\u2010coupled CTR as well as CTR alone. This means that although in binding assays human 2 receptors. Adrenomedullin 2/intermedin also has high affinity for the AM2 receptor [http://www.ncbi.nlm.nih.gov/pubmed/21658025?dopt=AbstractPlus]. CGRP\u2010(8\u201037) acts as an antagonist of CGRP (pKi 8) and inhibits some AM and AMY responses (pKi 6\u20107). It is weak at CT receptors. HumanAM\u2010(22\u201052)has some selectivity towardsAM receptors, but with modest potency (pKi 7), limiting its use [http://www.ncbi.nlm.nih.gov/pubmed/12970090?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=702 shows the greatest selectivity between receptors but still has significant affinity for AMY1 receptors [http://www.ncbi.nlm.nih.gov/pubmed/26125036?dopt=AbstractPlus].The ligands described have limited selectivity. Adrenomedullin has appreciable affinity for CGRP receptors. CGRP can show significant cross\u2010reactivity at AMY receptors and AMe.g. Ca2+, ERK, Akt), and G proteins can be activated [http://www.ncbi.nlm.nih.gov/pubmed/20633935?dopt=AbstractPlus]. There is evidence that CGRP\u2010RCP is important for the coupling of CLR to adenylyl cyclase [http://www.ncbi.nlm.nih.gov/pubmed/10903324?dopt=AbstractPlus].Gs is a prominent route for effector coupling for CLR and CTR but other pathways Update on the pharmacology of calcitonin/CGRP family of peptides: IUPHAR Review 25. Br. J. Pharmacol.175: 3\u201017 [https://www.ncbi.nlm.nih.gov/pubmed/29059473?dopt=AbstractPlus]Hay DL et al. (2014) Calcitonin gene\u2010related peptide: physiology and pathophysiology. Physiol. Rev.94: 1099\u2010142 [https://www.ncbi.nlm.nih.gov/pubmed/25287861?dopt=AbstractPlus]Russell FA et al. (2016) Receptor Activity\u2010Modifying Proteins (RAMPs): New Insights and Roles. Annu. Rev. Pharmacol. Toxicol.56: 469\u201087 [https://www.ncbi.nlm.nih.gov/pubmed/26514202?dopt=AbstractPlus]Hay DL Annu. Rev. Pharmacol. Toxicol.55: 533\u201052 [https://www.ncbi.nlm.nih.gov/pubmed/25340934?dopt=AbstractPlus]Russo AF. (2015) Calcitonin gene\u2010related peptide (CGRP): a new target for migraine. et al. (2015) Bench\u2010to\u2010bedside pharmacology of adrenomedullin. Eur. J. Pharmacol.764: 140\u20108 [https://www.ncbi.nlm.nih.gov/pubmed/26144371?dopt=AbstractPlus]Kato J NC\u2010IUPHARprovisional nomenclature as recommended by [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) responds to multiple endogenous ligands, including extracellular calcium and other divalent/trivalent cations, polyamines and polycationic peptides, L\u2010amino acids (particularly L\u2010Trp and L\u2010Phe), glutathione and various peptide analogues, ionic strength and extracellular pH (reviewed in [http://www.ncbi.nlm.nih.gov/pubmed/24111791?dopt=AbstractPlus]). While divalent/trivalent cations, polyamines and polycations are CaS receptor agonists , L\u2010amino acids, glutamyl peptides, ionic strength and pH are allosteric modulators of agonist function . Indeed, L\u2010amino acids have been identified as \"co\u2010agonists\", with both concomitant calcium and L\u2010amino acid binding required for full receptor activation . The sensitivity of the CaS receptor to primary agonists is increased by elevated extracellular pH [http://www.ncbi.nlm.nih.gov/pubmed/25556167?dopt=AbstractPlus] or decreased extracellular ionic strength [http://www.ncbi.nlm.nih.gov/pubmed/9677383?dopt=AbstractPlus]. This receptor bears no sequence or structural relation to the plant calcium receptor, also called CaS.The calcium\u2010sensing receptor or neonatal severe hyperparathyroidism [http://www.ncbi.nlm.nih.gov/pubmed/27647839?dopt=AbstractPlus] and in Casr null mice , which exhibit similar increases in PTH secretion and blood calcium levels. Gain\u2010of\u2010function CaS mutations are associated with autosomal dominant hypocalcaemia and Bartter syndrome type V [http://www.ncbi.nlm.nih.gov/pubmed/27647839?dopt=AbstractPlus].The CaS receptor has a number of physiological functions, but it is best known for its central role in parathyroid and renal regulation of extracellular calcium homeostasis , but in some cell types can couple to Gs [http://www.ncbi.nlm.nih.gov/pubmed/20032198?dopt=AbstractPlus]. However, the CaS receptor can form heteromers with Class C GABAB and mGlu1/5 receptors [http://www.ncbi.nlm.nih.gov/pubmed/11489900?dopt=AbstractPlus], which may introduce further complexity in its signalling capabilities.The CaS receptor primarily couples to Ghttp://www.ncbi.nlm.nih.gov/pubmed/21406038?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/24050279?dopt=AbstractPlus]. Further, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=8375 is a novel peptide positive allosteric modulator of the receptor [http://www.ncbi.nlm.nih.gov/pubmed/23674604?dopt=AbstractPlus]. Agonists and positive allosteric modulators of the CaS receptor are termed Type I and II calcimimetics, respectively, and can suppress parathyroid hormone ) secretion [http://www.ncbi.nlm.nih.gov/pubmed/9520489?dopt=AbstractPlus]. Negative allosteric modulators are called calcilytics and can act to increase http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1785, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:9606) secretion [http://www.ncbi.nlm.nih.gov/pubmed/11561095?dopt=AbstractPlus].Multiple other small molecule chemotypes are positive and negative allosteric modulators of the CaS receptor Brown EM. (2013) Role of the calcium\u2010sensing receptor in extracellular calcium homeostasis. et al. (2018) The calcium\u2010sensing receptor in physiology and in calcitropic and noncal citropic diseases. Nat Rev Endocrinol15: 33\u201051 [https://www.ncbi.nlm.nih.gov/pubmed/30443043?dopt=AbstractPlus]Hannan FM et al. (2013) Calcium\u2010sensing receptor (CaSR): pharmacological properties and signaling pathways. Best Pract. Res. Clin. Endocrinol. Metab.27: 315\u201031 [https://www.ncbi.nlm.nih.gov/pubmed/23856262?dopt=AbstractPlus]Conigrave AD et al. (2018) Discovery and Development of Calcimimetic and Calcilytic Compounds. Prog Med Chem57: 1\u201086 [https://www.ncbi.nlm.nih.gov/pubmed/29680147?dopt=AbstractPlus]Nemeth EF NC\u2010IUPHAR Subcommittee on Cannabinoid Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/21079038?dopt=AbstractPlus]) are activated by endogenous ligands that include N\u2010arachidonoylethanolamine (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2364), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5444\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5444, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5445 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=729. Potency determinations of endogenous agonists at these receptors are complicated by the possibility of differential susceptibility of endogenous ligands to enzymatic conversion [http://www.ncbi.nlm.nih.gov/pubmed/17876303?dopt=AbstractPlus].Cannabinoid receptors (1 and CB2 receptors [http://www.ncbi.nlm.nih.gov/pubmed/23108552?dopt=AbstractPlus]. Two of these medicines were developed to suppress nausea and vomiting produced by chemotherapy. These are http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=9071 (Cesamet\u00ae), a synthetic CB1/CB2 receptor agonist, and synthetic http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2424 , which can also be used as an appetite stimulant. The third medicine, Sativex\u00ae, contains mainly http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2424 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4150, both extracted from cannabis, and is used to treat multiple sclerosis and cancer pain.There are currently three licenced cannabinoid medicines each of which contains a compound that can activate CB1 and CB2 receptors may be labelled with . There is evidence for an allosteric site on the CB1 receptor [http://www.ncbi.nlm.nih.gov/pubmed/16113085?dopt=AbstractPlus]. All of the compounds listed as antagonists behave as inverse agonists in some bioassay systems [http://www.ncbi.nlm.nih.gov/pubmed/21079038?dopt=AbstractPlus]. For some cannabinoid receptor ligands, additional pharmacological targets that include GPR55 and GPR119 have been identified [http://www.ncbi.nlm.nih.gov/pubmed/21079038?dopt=AbstractPlus]. Moreover, http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=114 although showing little structural similarity to CB1 and CB2 receptors, respond to endogenous agents that are structurally similar to the endogenous cannabinoid ligands [http://www.ncbi.nlm.nih.gov/pubmed/21079038?dopt=AbstractPlus].Both CBet al. (2002) International Union of Pharmacology. XXVII. Classification of cannabinoid receptors. Pharmacol. Rev.54: 161\u2010202 [https://www.ncbi.nlm.nih.gov/pubmed/12037135?dopt=AbstractPlus]Howlett AC et al. (2010) International Union of Basic and Clinical Pharmacology. LXXIX. Cannabinoid receptors and their ligands: beyond CB_1 and CB_2. Pharmacol. Rev.62: 588\u2010631 [https://www.ncbi.nlm.nih.gov/pubmed/21079038?dopt=AbstractPlus]Pertwee RG Curr. Med. Chem.17: 1360\u201081 [https://www.ncbi.nlm.nih.gov/pubmed/20166927?dopt=AbstractPlus]Pertwee RG. (2010) Receptors and channels targeted by synthetic cannabinoid receptor agonists and antagonists. NC\u2010IUPHARrecommended by ). The chemoattractant protein and adipokine, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2945 , has been shown to be the endogenous ligand for both chemerin family receptors. Chemerin1 was the founding family member, and when GPR1 was de\u2010orphanised it was re\u2010named Chermerin2 [http://www.ncbi.nlm.nih.gov/pubmed/29279348?dopt=AbstractPlus]. Chemerin1 is also activated by the lipid\u2010derived, antiinflammatory ligand http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3333 (RvE1), which is formed via the sequential metabolism of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3362 by aspirin\u2010modified cyclooxygenase and lipoxygenase . In addition, two GPCRs for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3934 (RvD1) have been identified: FPR2/ALX, the lipoxin A4 receptor, and GPR32, an orphan receptor [http://www.ncbi.nlm.nih.gov/pubmed/20080636?dopt=AbstractPlus].Nomenclature for the chemerin receptors is presented as Ki=9.2 [http://www.ncbi.nlm.nih.gov/pubmed/27742615?dopt=AbstractPlus].CCX832 (structure not disclosed) is a selective antagonist, pet al. (2018) International Union of Basic and Clinical Pharmacology CIII: Chemerin Receptors CMKLR1 (Chemerin1) and GPR1 (Chemerin2) Nomenclature, Pharmacology, and Function. Pharmacol. Rev.70: 174\u2010196 [https://www.ncbi.nlm.nih.gov/pubmed/29279348?dopt=AbstractPlus]Kennedy AJ et al. (2018) Mechanisms and Functions of Chemerin in Cancer: Potential Roles in Therapeutic Intervention. Front Immunol9: 2772 [https://www.ncbi.nlm.nih.gov/pubmed/30555465?dopt=AbstractPlus]Shin WJ NC\u2010IUPHAR Subcommittee on Chemokine Receptorsnomenclature as agreed by the ) comprise a large subfamily of 7TM proteins that bind one or more chemokines, a large family of small cytokines typically possessing chemotactic activity for leukocytes. Additional hematopoietic and non\u2010hematopoietic roles have been identified for many chemokines in the areas of embryonic development, immune cell proliferation, activation and death, viral infection, and as antibiotics, among others. Chemokine receptors can be divided by function into two main groups: G protein\u2010coupled chemokine receptors, which mediate leukocyte trafficking, and \"Atypical chemokine receptors\", which may signal through non\u2010G protein\u2010coupled mechanisms and act as chemokine scavengers to downregulate inflammation or shape chemokine gradients [http://www.ncbi.nlm.nih.gov/pubmed/24218476?dopt=AbstractPlus].Chemokine receptors (n= 28), CXC and CX3C (n= 1) chemokines all have four conserved cysteines, with zero, one and three amino acids separating the first two cysteines respectively. C chemokines (n= 2) have only the second and fourth cysteines found in other chemokines. Chemokines can also be classified by function into homeostatic and inflammatory subgroups. Most chemokine receptors are able to bind multiple high\u2010affinity chemokine ligands, but the ligands for a given receptor are almost always restricted to the same structural subclass. Most chemokines bind to more than one receptor subtype. Receptors for inflammatory chemokines are typically highly promiscuous with regard to ligand specificity, and may lack a selective endogenous ligand. G protein\u2010coupled chemokine receptors are named acccording to the class of chemokines bound, whereas ACKR is the root acronym for atypical chemokine receptors [http://www.ncbi.nlm.nih.gov/pubmed/25958743?dopt=AbstractPlus]. There can be substantial cross\u2010species differences in the sequences of both chemokines and chemokine receptors, and in the pharmacology and biology of chemokine receptors. Endogenous and microbial non\u2010chemokine ligands have also been identified for chemokine receptors. Many chemokine receptors function as HIV co\u2010receptors, but CCR5 is the only one demonstrated to play an essential role in HIV/AIDS pathogenesis. The tables include bothstandard chemokine receptor names [http://www.ncbi.nlm.nih.gov/pubmed/10714678?dopt=AbstractPlus] and aliases.Chemokines in turn can be divided by structure into four subclasses by the number and arrangement of conserved cysteines. CC , but their role in viral life cycles is not established. Viruses can exploit or subvert the chemokine system by producing chemokine antagonists and scavengers. Three chemokine receptor antagonists have now been approved by the FDA: 1) the CCR5 antagonist http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=806 (Pfizer) for treatment of HIV/AIDS in patients with CCR5\u2010using strains; and 2) the CXCR4 antagonist http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=844 (Sanofi) for hematopoietic stem cell mobilization with http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4934 in patients undergoing transplantation in the context of chemotherapy for Hodgkins\u2019 Disease and multiple myeloma; and 3) the CCR4 blocking antibody Poteligeo for mycosis fungoides or Sezary syndrome.Specific chemokine receptors facilitate cell entry by microbes, such as ACKR1 for et al. (2015) An atypical addition to the chemokine receptor nomenclature: IUPHAR Review 15. Br. J. Pharmacol.172: 3945\u20109 [https://www.ncbi.nlm.nih.gov/pubmed/25958743?dopt=AbstractPlus]Bachelerie F et al. (2000) International Union of Pharmacology. XXII. Nomenclature for chemokine receptors. Pharmacol. Rev.52: 145\u2010176 [https://www.ncbi.nlm.nih.gov/pubmed/10699158?dopt=AbstractPlus]Murphy PM et al. (2012) Targeting chemokine receptors in chronic inflammatory diseases: an extensive review. Pharmacol. Ther.133: 1\u201018 [https://www.ncbi.nlm.nih.gov/pubmed/21839114?dopt=AbstractPlus]Koelink PJ et al. (2012) Pharmacological modulation of chemokine receptor function. Br. J. Pharmacol.165: 1617\u201043 [https://www.ncbi.nlm.nih.gov/pubmed/21699506?dopt=AbstractPlus]Scholten DJ Pharmacol. Rev.54: 227\u20109 [https://www.ncbi.nlm.nih.gov/pubmed/12037138?dopt=AbstractPlus]Murphy PM. (2002) International Union of Pharmacology. XXX. Update on chemokine receptor nomenclature. NC\u2010IUPHAR Subcommittee on CCK receptorsnomenclature as agreed by the , with some alternatively spliced forms most often identified in neoplastic cells. The CCK receptor subtypes are distinguished by their peptide selectivity, with the CCK1 receptor requiring the carboxyl\u2010terminal heptapeptide\u2010amide that includes a sulfated tyrosine for high affinity and potency, while the CCK2 receptor requires only the carboxyl\u2010terminal tetrapeptide shared by each CCK and gastrin peptides. These receptors have characteristic and distinct distributions, with both present in both the central nervous system and peripheral tissues.Cholecystokinin receptors region, has been described to be present particularly in certain neoplasms where mRNA mis\u2010splicing has been commonly observed [http://www.ncbi.nlm.nih.gov/pubmed/12429993?dopt=AbstractPlus], but it is not clear that this receptor splice form plays a special role in carcinogenesis. Another alternative splicing event for the CCK2 receptor was reported [http://www.ncbi.nlm.nih.gov/pubmed/8415658?dopt=AbstractPlus], with alternative donor sites in exon 4 resulting in long (452 amino acids) and short (447 amino acids) forms of the receptor differing by five residues in ICL3, however, no clear functional differences have been observed.While a cancer\u2010specific CCK receptor has been postulated to exist, which also might be responsive to incompletely processed forms of CCK (Gly\u2010extended forms), this has never been isolated. An alternatively spliced form of the CCKRev Neurosci28: 573\u2010585 [https://www.ncbi.nlm.nih.gov/pubmed/28343167?dopt=AbstractPlus]Ballaz S. (2017) The unappreciated roles of the cholecystokinin receptor CCK(1) in brain functioning. Regul. Pept.155: 6\u201010 [https://www.ncbi.nlm.nih.gov/pubmed/19345244?dopt=AbstractPlus]Dockray GJ. (2009) Cholecystokinin and gut\u2010brain signalling. et al. (2010) Therapeutic potential for novel drugs targeting the type 1 cholecystokinin receptor. Br. J. Pharmacol.159: 1009\u201021 [https://www.ncbi.nlm.nih.gov/pubmed/19922535?dopt=AbstractPlus]Cawston EE et al. (2006) Cholecystokinin and gastrin receptors. Physiol. Rev.86: 805\u201047 [https://www.ncbi.nlm.nih.gov/pubmed/16816139?dopt=AbstractPlus]Dufresne M NC\u2010IUPHAR subcommittee on the Class Frizzled GPCRsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/21079039?dopt=AbstractPlus]), are GPCRs originally identified in Drosophila [http://www.ncbi.nlm.nih.gov/pubmed/1334084?dopt=AbstractPlus], which are highly conserved across species. While SMO shows structural resemblance to the 10 FZDs, it is functionally separated as it mediates effects in the Hedgehog signaling pathway [http://www.ncbi.nlm.nih.gov/pubmed/21079039?dopt=AbstractPlus]. FZDs are activated by WNTs, which are cysteine\u2010rich lipoglycoproteins withfundamentalfunctions inontogeny and tissue homeostasis. FZD signalling was initially divided into two pathways, being either dependent on the accumulation of the transcription regulator http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5371 or being \u03b2\u2010catenin\u2010independent . WNT stimulation of FZDs can, in cooperation with the low density lipoprotein receptors https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:6697 (http://www.uniprot.org/uniprot/O75197) and https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:6698 (http://www.uniprot.org/uniprot/O75581), lead to the inhibition of a constitutively active destruction complex, which results in the accumulation of \u03b2\u2010catenin and subsequently its translocation to the nucleus. \u03b2\u2010Catenin, in turn, modifies gene transcription by interacting with TCF/LEF transcription factors. \u03b2Catenin\u2010independent FZD signalling is far more complex with regard to the diversity of the activated pathways. WNT/FZD signalling can lead to the activation of heterotrimeric G proteins , the elevation of intracellular calcium [http://www.ncbi.nlm.nih.gov/pubmed/9389482?dopt=AbstractPlus], activation of cGMP\u2010specific PDE6 [http://www.ncbi.nlm.nih.gov/pubmed/12471263?dopt=AbstractPlus] and elevation of cAMP as well as RAC\u20101, JNK, Rho and Rho kinase signalling [http://www.ncbi.nlm.nih.gov/pubmed/19651774?dopt=AbstractPlus]. Novel resonance energy transfer\u2010based tools have allowed the study of the GPCR\u2010like nature of FZDs in greater detail. Upon ligand stimulation, FZDs undergo conformational changes and signal via heterotrimeric G proteins . Furthermore, the phosphoprotein Dishevelled constitutes a key player in WNT/FZD signalling. Importantly, FZDs exist in at least two distinct conformational states that regulate the pathway selection [http://www.ncbi.nlm.nih.gov/pubmed/30737406?dopt=AbstractPlus]. As with other GPCRs, members of the Frizzled family are functionally dependent on the arrestin scaffolding protein for internalization [http://www.ncbi.nlm.nih.gov/pubmed/12958365?dopt=AbstractPlus], as well as for \u03b2\u2010catenin\u2010dependent [http://www.ncbi.nlm.nih.gov/pubmed/17426148?dopt=AbstractPlus] and \u2010independent signalling. The pattern of cell signalling is complicated by the presence of additional ligands, which can enhance or inhibit FZD signalling (secreted Frizzled\u2010related proteins (sFRP), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5372 (WIF), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3704 or Dickkopf (DKK)), as well as modulatory (co)\u2010receptors with http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=304#Type XV RTKs: RYK, http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=304#Type VIII RTKs: ROR1, http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=304#Type VIII RTKs: ROR2 and Kremen, which may also function as independent signalling proteins.Receptors of the Class Frizzled , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3673 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3674 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3675 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3549 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3547 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3548 (pEC50 7.7\u20108.9 [http://www.ncbi.nlm.nih.gov/pubmed/30514810?dopt=AbstractPlus]), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3676 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3678 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3679 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3681 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3682 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3683 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3684 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3686 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3687, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3688 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3689 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3690 .Extracellular proteins that interact with FZDs:http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1063 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3700 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3691 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3692 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3693 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3694 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3695 .Extracellular proteins that interact with WNTs or LRPs:http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3701 , https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:18081 (http://www.uniprot.org/uniprot/Q9Y5W5), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3704 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3702 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3703 Small exogenous ligands: Foxy\u20105 [http://www.ncbi.nlm.nih.gov/pubmed/18927296?dopt=AbstractPlus], Box\u20105 [http://www.ncbi.nlm.nih.gov/pubmed/19901340?dopt=AbstractPlus], UM206 [http://www.ncbi.nlm.nih.gov/pubmed/21931076?dopt=AbstractPlus], and XWnt8 (http://www.uniprot.org/uniprot/P28026) also known as mini\u2010Wnt8.Ligands associated with SMO signalling:http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2718, oxysterols .et al. (2009) Proximal events in Wnt signal transduction. Nat. Rev. Mol. Cell Biol.10: [https://www.ncbi.nlm.nih.gov/pubmed/22935904?dopt=AbstractPlus]Angers S Cold Spring Harb Perspect Biol4: 468\u201077 [https://www.ncbi.nlm.nih.gov/pubmed/19536106?dopt=AbstractPlus]van Amerongen R. (2012) Alternative Wnt pathways and receptors. https://www.ncbi.nlm.nih.gov/pubmed/26969975?dopt=AbstractPlus]Schulte G.(2015)Frizzleds and WNT/\u03b2\u2010cateninsignaling\u2013The black boxof ligand\u2010receptor selectivity, 113\u201039 [et al. (2016) Frizzled Receptors in Development and Disease. Curr. Top. Dev. Biol.117: complex stoichiometry and activation kinetics. Eur. J. Pharmacol.763: 191\u20105 [https://www.ncbi.nlm.nih.gov/pubmed/26003275?dopt=AbstractPlus]Wang Y et al. (2018) Frizzleds as GPCRs \u2010 More Conventional Than We Thought! Trends Pharmacol. Sci.39: 828\u2010842 [https://www.ncbi.nlm.nih.gov/pubmed/30049420?dopt=AbstractPlus]Schulte G NC\u2010IUPHAR subcommittee on Complement peptide receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/23383423?dopt=AbstractPlus]) are activated by the endogenous 75 amino\u2010acid anaphylatoxin polypeptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3640 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=573 , generated upon stimulation of the complement cascade. C3a and C5a exert their functions through binding to their receptors (C3aR and C5aR), causing cell activation and triggering cellular degranulation that contributes to the local inflammation.Complement peptide receptors binds [125I]C5a with no clear signalling function, but has a putative role opposing inflammatory responses . Binding to this site may be displaced with the rank order http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=574 (https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:1331)> http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=573 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5367 to compete . C5a2 appears to lack G protein signalling and has been termed a decoy receptor [http://www.ncbi.nlm.nih.gov/pubmed/19100624?dopt=AbstractPlus]. However, C5a2 does recruit arrestin after ligand binding, which might provide a signaling pathway for this receptor , and forms heteromers with C5a1. C5a, but not C5a\u2010des Arg, induces upregulation of heteromer formation between complement C5a receptors C5a1 and C5a2 [http://www.ncbi.nlm.nih.gov/pubmed/24060963?dopt=AbstractPlus]. There are also reports of pro\u2010inflammatory activity of C5a2, mediated by HMGB1, but the signaling pathway that underlies this is currently unclear (reviewed in [http://www.ncbi.nlm.nih.gov/pubmed/23239822?dopt=AbstractPlus]). More recently, work in T cells has shown that C5a1 and C5a2 act in opposition to each other and that altering the equilibrium between the two receptors, by differential expression or production of C5a\u2010des Arg (which favours C5a2), can affect the final cellular response [http://www.ncbi.nlm.nih.gov/pubmed/27313051?dopt=AbstractPlus].et al. (2016) A novel \"complement\u2010metabolism\u2010inflammasome axis\" as a key regulator of immune cell effector function. Eur. J. Immunol.46: 1563\u201073 [https://www.ncbi.nlm.nih.gov/pubmed/27184294?dopt=AbstractPlus]Arbore G et al. (2018) Complement C3a receptor modulates embryonic neural progenitor cell proliferation and cognitive performance. Mol. Immunol.101: 176\u2010181 [https://www.ncbi.nlm.nih.gov/pubmed/30449309?dopt=AbstractPlus]Coulthard LG et al. (2017) Novel insights into the expression pattern of anaphylatoxin receptors in mice and men. Mol. Immunol.89: 44\u201058 [https://www.ncbi.nlm.nih.gov/pubmed/28600003?dopt=AbstractPlus]Laumonnier Y et al. (2013) C5L2: a controversial receptor of complement anaphylatoxin, C5a. FASEB J.27: 855\u201064 [https://www.ncbi.nlm.nih.gov/pubmed/23239822?dopt=AbstractPlus]Li R et al. (2007) Function, structure and therapeutic potential of complement C5a receptors. Br. J. Pharmacol.152: 429\u201048 [https://www.ncbi.nlm.nih.gov/pubmed/17603557?dopt=AbstractPlus]Monk PN et al. (2018) Intracellular complement activation\u2010An alarm raising mechanism? Semin. Immunol.38: 54\u201062 [https://www.ncbi.nlm.nih.gov/pubmed/29631809?dopt=AbstractPlus]Reichhardt MP NC\u2010IUPHAR subcommittee on Corticotropin\u2010releasing Factor Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/12615952?dopt=AbstractPlus]) receptors are activated by the endogenous peptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=912 , a 41 aminoacid peptide, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=919 , 40 amino\u2010acids, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=921 , 38 amino\u2010acids and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=928 , 38 amino\u2010acids. CRF1 and CRF2 receptors are activated non\u2010selectively by CRH and UCN. CRF2 receptors are selectively activated by UCN2 and UCN3. Binding to CRF receptors can be conducted using radioligands [125I]Tyr0\u2010CRF or .A CRF binding protein has been identified The Corticotropin\u2010Releasing Factor Family: Physiology of the Stress Response. Physiol. Rev.98: 2225\u20102286 [https://www.ncbi.nlm.nih.gov/pubmed/30109816?dopt=AbstractPlus]Deussing JM et al. (2003) International Union of Pharmacology. XXXVI. Current status of the nomenclature for receptors for corticotropin\u2010releasing factor and their ligands. Pharmacol Rev.55: 21\u201026 [https://www.ncbi.nlm.nih.gov/pubmed/12615952?dopt=AbstractPlus]Hauger RL Br. J. Pharmacol.166: 85\u201097 [https://www.ncbi.nlm.nih.gov/pubmed/21883143?dopt=AbstractPlus]Grammatopoulos DK. (2012) Insights into mechanisms of corticotropin\u2010releasing hormone receptor signal transduction. et al. (2011) Members of CRF family and their receptors: from past to future. Curr. Med. Chem.18: 2583\u2010600 [https://www.ncbi.nlm.nih.gov/pubmed/21568890?dopt=AbstractPlus]Liapakis G et al. (2016) Corticotropin\u2010Releasing Factor Receptors and Their Interacting Proteins: Functional Consequences. Mol. Pharmacol.90: 627\u2010632 [https://www.ncbi.nlm.nih.gov/pubmed/27612874?dopt=AbstractPlus]Slater PG et al. (2017) Structures of the First Extracellular Domain of CRF Receptors. Curr Mol Pharmacol10: 318\u2010324 [https://www.ncbi.nlm.nih.gov/pubmed/28103782?dopt=AbstractPlus]Zelenay V NC\u2010IUPHAR Subcommittee on Dopamine Receptorsnomenclature as agreed by the Beaulieu JM et al. (2011) The physiology, signaling, and pharmacology of dopamine receptors. Pharmacol. Rev.63: 182\u2010217 [https://www.ncbi.nlm.nih.gov/pubmed/21303898?dopt=AbstractPlus]Beaulieu JM Synapse65: 892\u2010909 [https://www.ncbi.nlm.nih.gov/pubmed/21308799?dopt=AbstractPlus]Cumming P. (2011) Absolute abundances and affinity states of dopamine receptors in mammalian brain: A review. et al. (2010) Dopamine D2\u2010D3 receptor heteromers: pharmacological properties and therapeutic significance. Curr Opin Pharmacol10: 100\u20107 [https://www.ncbi.nlm.nih.gov/pubmed/19896900?dopt=AbstractPlus]Maggio R et al. (2011) Dopamine D4 receptor gene DRD4 and its association with psychiatric disorders. Med. Sci. Monit.17: RA215\u201020 [https://www.ncbi.nlm.nih.gov/pubmed/21873960?dopt=AbstractPlus]Pt\u00e1cek R et al. (1998) Dopamine Receptors. In The IUPHAR Compendium of Receptor Characterization and Classification Edited by Girdlestone D: IUPHAR Media: 141\u2010151Schwartz J\u2010C Pharmacol. Ther.128: 37\u201060 [https://www.ncbi.nlm.nih.gov/pubmed/20547182?dopt=AbstractPlus]Undieh AS. (2010) Pharmacology of signaling induced by dopamine D(1)\u2010like receptor activation. NC\u2010IUPHAR Subcommittee on Endothelin Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/12037137?dopt=AbstractPlus]) are activated by the endogenous 21 amino\u2010acid peptides endothelins 1\u20103 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=990 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1004 ).Endothelin receptors and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3886 [http://www.ncbi.nlm.nih.gov/pubmed/30413709?dopt=AbstractPlus] have been reported.Splice variants of the ETet al. (2013) Endothelin receptor antagonists. Handb Exp Pharmacol218: 199\u2010227 https://www.ncbi.nlm.nih.gov/pubmed/24092342?dopt=AbstractPlusClozel M Pharmacol. Rev.54: 219\u201026 https://www.ncbi.nlm.nih.gov/pubmed/12037137?dopt=AbstractPlusDavenport AP. (2002) International Union of Pharmacology. XXIX. Update on endothelin receptor nomenclature. et al. (2016) Endothelin. Pharmacol. Rev.68: 357\u2010418 https://www.ncbi.nlm.nih.gov/pubmed/26956245?dopt=AbstractPlusDavenport AP et al. (2018) New drugs and emerging therapeutic targets in the endothelin signaling pathway and prospects for personalized precision medicine. Physiol Res67: S37\u2010S54 https://www.ncbi.nlm.nih.gov/pubmed/29947527?dopt=AbstractPlusDavenport AP et al. (2014) Endothelin@25 \u2010 new agonists, antagonists, inhibitors and emerging research frontiers: IUPHAR Review 12. Br. J. Pharmacol.171: 5555\u201072 https://www.ncbi.nlm.nih.gov/pubmed/25131455?dopt=AbstractPlusMaguire JJ NC\u2010IUPHAR Subcommittee on the G protein\u2010coupled estrogen receptornomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/26023144?dopt=AbstractPlus]) was identified following observations of estrogen\u2010evoked http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2352 signalling in breast cancer cells [http://www.ncbi.nlm.nih.gov/pubmed/8078914?dopt=AbstractPlus], which mirrored the differential expression of an orphan 7\u2010transmembrane receptor GPR30 [http://www.ncbi.nlm.nih.gov/pubmed/9367686?dopt=AbstractPlus]. There are observations of both cell\u2010surface and intracellular expression of the GPER receptor . Selective agonist/ antagonists for GPER have been characterized [http://www.ncbi.nlm.nih.gov/pubmed/26023144?dopt=AbstractPlus]. Antagonists of the nuclear estrogen receptor, such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1015 [http://www.ncbi.nlm.nih.gov/pubmed/11043579?dopt=AbstractPlus], http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1016 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2820 [http://www.ncbi.nlm.nih.gov/pubmed/24379833?dopt=AbstractPlus], as well as the flavonoid \u2019phytoestrogens\u2019 http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2826 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5346 [http://www.ncbi.nlm.nih.gov/pubmed/15090535?dopt=AbstractPlus], are agonists of GPER. A complete review of GPER pharmacology has been recently published [http://www.ncbi.nlm.nih.gov/pubmed/26023144?dopt=AbstractPlus]. The roles of GPER in physiological systems throughout the body and in cancer have also been reviewed .The G protein\u2010coupled estrogen receptor Twenty years of the G protein\u2010coupled estrogen receptor GPER: Historical and personal perspectives. J. Steroid Biochem. Mol. Biol.176: 4\u201015 https://www.ncbi.nlm.nih.gov/pubmed/28347854?dopt=AbstractPlusBarton M et al. (2015) The G\u2010protein coupled estrogen receptor, GPER: The inside and inside\u2010out story. Mol. Cell. Endocrinol.418 Pt 3: 207\u201019 https://www.ncbi.nlm.nih.gov/pubmed/26190834?dopt=AbstractPlusGaudet HM et al. (2015) International Union of Basic and Clinical Pharmacology. XCVII. G Protein\u2010Coupled Estrogen Receptor and Its Pharmacologic Modulators. Pharmacol. Rev.67: 505\u201040 https://www.ncbi.nlm.nih.gov/pubmed/26023144?dopt=AbstractPlusProssnitz ER et al. (2015) What have we learned about GPER function in physiology and disease from knockout mice? J. Steroid Biochem. Mol. Biol.153: 114\u201026 https://www.ncbi.nlm.nih.gov/pubmed/26189910?dopt=AbstractPlusProssnitz ER http://www.ensembl.org/Homo_sapiens/Gene/Family/Genes?family=ENSFM00510000502765 (NC\u2010IUPHAR Subcommittee on the formylpeptide receptor familynomenclature agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/19498085?dopt=AbstractPlus]) respond to exogenous ligands such as the bacterial product http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1022 (fMLP) and endogenous ligands such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1031 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3570 , amyloid \u03b242, serum amyloid A and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1026, derived from http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5370 .The http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=35 receptor page.Note that the data for FPR2/ALX are also reproduced on the et al. (2015) The Role of Formylated Peptides and Formyl Peptide Receptor 1 in Governing Neutrophil Function during Acute Inflammation. Am. J. Pathol.185: 1172\u20101184 https://www.ncbi.nlm.nih.gov/pubmed/25791526?dopt=AbstractPlusDorward DA et al. (2010) Therapeutic anti\u2010inflammatory potential of formyl\u2010peptide receptor agonists. Pharmacol. Ther.127: 175\u201088 https://www.ncbi.nlm.nih.gov/pubmed/20546777?dopt=AbstractPlusDufton N et al. (2012) G protein\u2010coupled receptor FPR1 as a pharmacologic target in inflammation and human glioblastoma. Int. Immunopharmacol.14: 283\u20108 https://www.ncbi.nlm.nih.gov/pubmed/22863814?dopt=AbstractPlusLiu M et al. (2011) N\u2010formyl peptide receptor 3 (FPR3) departs from the homologous FPR2/ALX receptor with regard to the major processes governing chemoattractant receptor regulation, expression at the cell surface, and phosphorylation. J. Biol. Chem.286: 26718\u201031 https://www.ncbi.nlm.nih.gov/pubmed/21543323?dopt=AbstractPlusRabiet MJ et al. (2012) Anti\u2010inflammatory drugs, eicosanoids and the annexin A1/FPR2 anti\u2010inflammatory system. Prostaglandins Other Lipid Mediat.98: 94\u2010100 https://www.ncbi.nlm.nih.gov/pubmed/22123264?dopt=AbstractPlusYazid S et al. (2009) International Union of Basic and Clinical Pharmacology. LXXIII. Nomenclature for the formyl peptide receptor (FPR) family. Pharmacol. Rev.61: 119\u201061 https://www.ncbi.nlm.nih.gov/pubmed/19498085?dopt=AbstractPlusYe RD NC\u2010IUPHAR Subcommittee on free fatty acid receptorsnomenclature as agreed by the ) are activated by free fatty acids. Long\u2010chain saturated and unsaturated fatty acids (including C14.0 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2806), C16:0 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1055), C18:1 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1054), C18:2 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1052), C18:3, (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1049), C20:4 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2391), C20:5,n\u20103 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3362) and C22:6,n\u20103 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1051)) activate FFA1 and FFA4 receptors , while short chain fatty acids (C2 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1058), C3 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1062), C4 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1059) and C5 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1061)) activate FFA2 and FFA3 receptors. The crystal structure for agonist bound FFA1 has been described [http://www.ncbi.nlm.nih.gov/pubmed/25043059?dopt=AbstractPlus].Free fatty acid receptors and long (377 amino acids) splice variants of human FFA4 have been reported . https://www.genenames.org/data/gene\u2010symbol\u2010report/%23!/hgnc_id/HGNC:4535 is a structurally\u2010unrelated G protein\u2010coupled receptor which has been found to respond to medium chain fatty acids [http://www.ncbi.nlm.nih.gov/pubmed/16966319?dopt=AbstractPlus].et al. (2016) The Pharmacology and Function of Receptors for Short\u2010Chain Fatty Acids. Mol. Pharmacol.89: 388\u201098 https://www.ncbi.nlm.nih.gov/pubmed/26719580?dopt=AbstractPlusBolognini D et al. (2013) The fatty acid receptor FFA1/GPR40 a decade later: how much do we know? Trends Endocrinol. Metab.24: 398\u2010407 https://www.ncbi.nlm.nih.gov/pubmed/23631851?dopt=AbstractPlusMancini AD et al. (2017) Complex Pharmacology of Free Fatty Acid Receptors. Chem. Rev.117: 67\u2010110 https://www.ncbi.nlm.nih.gov/pubmed/27299848?dopt=AbstractPlusMilligan G Biochem. Pharmacol.110\u2010111: 1\u201015 https://www.ncbi.nlm.nih.gov/pubmed/26827942?dopt=AbstractPlusMoniri NH. (2016) Free\u2010fatty acid receptor\u20104 (GPR120): Cellular and molecular function and its role in metabolic disorders. et al. (2008) International Union of Pharmacology. LXXI. Free fatty acid receptors FFA1, \u20102, and \u20103: pharmacology and pathophysiological functions. Pharmacol. Rev.60: 405\u201017 [https://www.ncbi.nlm.nih.gov/pubmed/19047536?dopt=AbstractPlus]Stoddart LA et al. (2014) Treatment of type 2 diabetes by free Fatty Acid receptor agonists. Front Endocrinol (Lausanne)5: 137 [https://www.ncbi.nlm.nih.gov/pubmed/25221541?dopt=AbstractPlus]Watterson KR B receptors are formed from the heterodimerization of two similar 7TM subunits termed GABAB1 and GABAB2 . GABAB receptors are widespread in the CNS and regulate both pre\u2010 and postsynaptic activity. The GABAB1 subunit, when expressed alone, binds both antagonists and agonists, but the affinity of the latter is generally 10\u2010100fold less than for the native receptor. Co\u2010expression of GABAB1 and GABAB2 subunits allows transport of GABAB1 to the cell surface and generates a functional receptor that can couple to signal transduction pathways such as high\u2010voltage\u2010activated Ca2+ channels , or inwardly rectifying potassium channels (Kir3) . The GABAB1 subunit harbours the GABA (orthosteric)\u2010binding site within an extracellular domain (ECD) venus flytrap module (VTM), whereas the GABAB2 subunit mediates G protein\u2010coupled signalling . The two subunits interact by direct allosteric coupling [http://www.ncbi.nlm.nih.gov/pubmed/21063387?dopt=AbstractPlus], such that GABAB2 increases the affinity of GABAB1 for agonists and reciprocally GABAB1 facilitates the coupling of GABAB2 to G proteins . GABAB1 and GABAB2 subunits assemble in a 1:1 stoichiometry by means of a coiled\u2010coil interaction between \u03b1\u2010helices within their carboxy\u2010termini that masks an endoplasmic reticulum retention motif (RXRR) within the GABAB1 subunit but other domains of the proteins also contribute to their heteromerization . Recent evidence indicates that higher order assemblies of GABAB receptor comprising dimers of heterodimers occur in recombinant expression systems and in vivo and that such complexes exhibit negative functional cooperativity between heterodimers . Adding further complexity, KCTD (potassium channel tetramerization proteins) 8, 12, 12b and 16 associate as tetramers with the carboxy terminus of the GABAB2 subunit to impart altered signalling kinetics and agonist potency to the receptor complex and are reviewed by [http://www.ncbi.nlm.nih.gov/pubmed/20655485?dopt=AbstractPlus]. The molecular complexity of GABAB receptors is further increased through association with trafficking and effector proteins and reviewed by [http://www.ncbi.nlm.nih.gov/pubmed/27905440?dopt=AbstractPlus]. Four isoforms of the human GABAB1 subunit have been cloned. The predominant GABAB1a and GABAB1b isoforms, which are most prevalent in neonatal and adult brain tissue respectively, differ in their ECD sequences as a result of the use of alternative transcription initiation sites. GABAB1a\u2010containing heterodimers localise todistal axonsand mediate inhibition of glutamate release in the CA3\u2010CA1 terminals, and GABA release onto the layer 5 pyramidal neurons, whereas GABAB1b\u2010containing receptors occur within dendritic spines and mediate slow postsynaptic inhibition . Only the 1a and 1b variants are identified as components of native receptors [http://www.ncbi.nlm.nih.gov/pubmed/12037141?dopt=AbstractPlus]. Additional GABAB1 subunit isoforms have been described in rodents and humans [http://www.ncbi.nlm.nih.gov/pubmed/21124972?dopt=AbstractPlus] and reviewed by [http://www.ncbi.nlm.nih.gov/pubmed/15269338?dopt=AbstractPlus].Functional GABASubunits50 values for the inhibition of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5381 binding to rat cerebral cortex membranes, are from . CGP27492 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1081), CGP35024 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1080) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1082 act as antagonists at human GABAA\u03c11 receptors, with potencies in the low micromolar range [http://www.ncbi.nlm.nih.gov/pubmed/21428811?dopt=AbstractPlus]. In addition to the ligands listed in the table, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=707 binds to the VTM of the GABAB1 subunit to act as a positive allosteric modulator of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1067 [http://www.ncbi.nlm.nih.gov/pubmed/10692480?dopt=AbstractPlus]. Synthetic positive allosteric modulators with low, or no, intrinsic activity include http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1079, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5446, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5503 [http://www.ncbi.nlm.nih.gov/pubmed/21181127?dopt=AbstractPlus] and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5504 . The site of action of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1079 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5446 appears to be on the heptahelical domain of the GABAB2 subunit . In the presence of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1079 or http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5446, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1069 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1068 behave as partial agonists [http://www.ncbi.nlm.nih.gov/pubmed/21428811?dopt=AbstractPlus]. A negative allosteric modulator of GABAB activity has been reported [http://www.ncbi.nlm.nih.gov/pubmed/25050158?dopt=AbstractPlus]. Knock\u2010out of the GABAB1 subunit in C57B mice causes the development of severe tonic\u2010clonic convulsions that prove fatal within a month of birth, whereas GABAB1\u2010/\u2010 BALB/c mice, although also displaying spontaneous epileptiform activity, are viable. The phenotype of the latter animals additionally includes hyperalgesia, hyperlocomotion , hyperdopaminergia, memory impairment and behaviours indicative of anxiety . A similar phenotype has been found for GABAB2\u2010/\u2010 BALB/c mice [http://www.ncbi.nlm.nih.gov/pubmed/15240800?dopt=AbstractPlus].Potencies of agonists and antagonists listed in the table, quantified as ICet al. (2002) International Union of Pharmacology. XXXIII. Mammalian gammaaminobutyricacid(B) receptors: structure and function. Pharmacol Rev.54: 247\u2010264 [https://www.ncbi.nlm.nih.gov/pubmed/12037141?dopt=AbstractPlus]Bowery NG Future Med Chem3: 163\u201075 [https://www.ncbi.nlm.nih.gov/pubmed/21428811?dopt=AbstractPlus]Froestl W. (2011) An historical perspective on GABAergic drugs. et al. (2012) Regulation of neuronal GABA(B) receptor functions by subunit composition. Nat. Rev. Neurosci.13: 380\u201094 [https://www.ncbi.nlm.nih.gov/pubmed/22595784?dopt=AbstractPlus]Gassmann M et al. (2016) Organization and functions of mGlu and GABAB receptor complexes. Nature540: 60\u201068 [https://www.ncbi.nlm.nih.gov/pubmed/27905440?dopt=AbstractPlus]Pin JP NC\u2010IUPHARprovisional nomenclature as recommended by [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) are activated by the endogenous peptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3592 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3594 . Human http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3592 is a 30 amino\u2010acid non\u2010amidated peptide [http://www.ncbi.nlm.nih.gov/pubmed/1714839?dopt=AbstractPlus]; in other species, it is 29 amino acids long and C\u2010terminally amidated. Amino acids 1\u201314 of galanin are highly conserved in mammals, birds, reptiles, amphibia and fish. Shorter peptide species and N\u2010terminally extended forms have been reported.Galanin receptors (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5356) is a high\u2010affinity agonist at GAL1/GAL2 (pKi 9), and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5357) is selective for GAL2 and GAL3 compared with GAL1 [http://www.ncbi.nlm.nih.gov/pubmed/15944007?dopt=AbstractPlus]. [125I]\u2010[Tyr26]galanin binds to all three subtypes with Kd values generally reported to range from 0.05 to 1 nM, depending on the assay conditions used . Porcine galanin\u2010(3\u201029) does not bind to cloned GAL1, GAL2 or GAL3 receptors, but a receptor that is functionally activated by porcine galanin\u2010(3\u201329) has been reported in pituitary and gastric smooth muscle cells . Additional galanin receptor subtypes are also suggested from studies with chimeric peptides , which act as antagonists in functional assays in the cardiovascular system [http://www.ncbi.nlm.nih.gov/pubmed/7693918?dopt=AbstractPlus], spinal cord [http://www.ncbi.nlm.nih.gov/pubmed/1373497?dopt=AbstractPlus], locus coeruleus, hippocampus [http://www.ncbi.nlm.nih.gov/pubmed/1720557?dopt=AbstractPlus] and hypothalamus , but exhibit agonist activity at some peripheral sites . The chimeric peptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3896, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3897, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3898, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3899 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3861 are agonists at GAL1 receptors expressed endogenously in Bowes human melanoma cells [http://www.ncbi.nlm.nih.gov/pubmed/10601261?dopt=AbstractPlus], and at heterologously expressed recombinant GAL1, GAL2 and GAL3 receptors . Recent studies have described the synthesis of a series of novel, systemically\u2010active, galanin analogues, with modest preferential binding at the GAL2 receptor. Specific chemical modifications to the galanin backbone increased brain levels of these peptides after i.v. injection and several of these peptides exerted a potent antidepressant\u2010like effect in mouse models of depression [http://www.ncbi.nlm.nih.gov/pubmed/23600864?dopt=AbstractPlus].et al. (2005) International Union of Pharmacology. XLVI. G protein\u2010coupled receptor list. Pharmacol Rev57: 279\u2010288 [https://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]Foord SM et al. (2015) Physiology, signaling, and pharmacology of galanin peptides and receptors: three decades of emerging diversity. Pharmacol. Rev.67: 118\u201075 [https://www.ncbi.nlm.nih.gov/pubmed/25428932?dopt=AbstractPlus]Lang R et al. (2011) The galanin peptide family in inflammation. Neuropeptides45: 1\u20108 [https://www.ncbi.nlm.nih.gov/pubmed/21087790?dopt=AbstractPlus]Lang R et al. (2011) Galanin\u2010like peptide (GALP) is a hypothalamic regulator of energy homeostasis and reproduction. Front Neuroendocrinol32: 1\u20109 [https://www.ncbi.nlm.nih.gov/pubmed/20558195?dopt=AbstractPlus]Lawrence C et al. (2012) Galanin receptors and ligands. Front Endocrinol (Lausanne)3: 146 [https://www.ncbi.nlm.nih.gov/pubmed/23233848?dopt=AbstractPlus]Webling KE NC\u2010IUPHAR Subcommittee for the Ghrelin receptornomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/16382107?dopt=AbstractPlus]) is activated by a 28 amino\u2010acid peptide originally isolated from rat stomach, where it is cleaved from a 117 aminoacid precursor . The human gene encoding the precursor peptide has 83% sequence homology to rat preproghrelin, although the mature peptides from rat and human differ by only two amino acids [http://www.ncbi.nlm.nih.gov/pubmed/11549267?dopt=AbstractPlus]. Alternative splicing results in the formation of a second peptide, [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3600 with equipotent biological activity [http://www.ncbi.nlm.nih.gov/pubmed/10801861?dopt=AbstractPlus]. A unique post\u2010translational modification [http://www.ncbi.nlm.nih.gov/pubmed/18267071?dopt=AbstractPlus] occurs in both peptides, essential for full activity in binding to ghrelin receptors in the hypothalamus and pituitary, and for the release of growth hormone from the pituitary [http://www.ncbi.nlm.nih.gov/pubmed/10604470?dopt=AbstractPlus]. Structure activity studies showed the first five N\u2010terminal amino acids to be the minimum required for binding [http://www.ncbi.nlm.nih.gov/pubmed/11087562?dopt=AbstractPlus], and receptor mutagenesis has indicated overlap of the ghrelin binding site with those for small molecule agonists and allosteric modulators of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1099 function [http://www.ncbi.nlm.nih.gov/pubmed/18923064?dopt=AbstractPlus]. In cell systems, the ghrelin receptor is constitutively active [http://www.ncbi.nlm.nih.gov/pubmed/15383539?dopt=AbstractPlus], but this is abolished by a naturally occurring mutation (A204E) that results in decreased cell surface receptor expression and is associated with familial short stature [http://www.ncbi.nlm.nih.gov/pubmed/16511605?dopt=AbstractPlus].The ghrelin receptor has been shown to bind (as [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3810) and have effects in the cardiovascular system [http://www.ncbi.nlm.nih.gov/pubmed/12969753?dopt=AbstractPlus], which raises the possible existence of different receptor subtypes in peripheral tissues and the central nervous system. A potent inverse agonist has been identified . http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3535, described as a ghrelin receptor agonist (pKi 7.8 and pD2 7.5 at human recombinant ghrelin receptors), has been shown to stimulate ghrelin receptor mediated food intake and gastric emptying but not elicit release of growth hormone, or modify ghrelin stimulated growth hormone release, thus pharmacologically discriminating the orexigenic and gastrointestinal actions of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1099 from the release of growth hormone [http://www.ncbi.nlm.nih.gov/pubmed/18719021?dopt=AbstractPlus]. A number of selective antagonists have been reported, including peptidomimetic [http://www.ncbi.nlm.nih.gov/pubmed/22798076?dopt=AbstractPlus] and non\u2010peptide small molecules including http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5870 .[Trends Neurosci.34: 31\u201040 [https://www.ncbi.nlm.nih.gov/pubmed/21035199?dopt=AbstractPlus]Andrews ZB. (2011) The extra\u2010hypothalamic actions of ghrelin on neuronal function. et al. (2010) Current and potential roles of ghrelin in clinical practice. J. Endocrinol. Invest.33: 823\u201038 [https://www.ncbi.nlm.nih.gov/pubmed/21293171?dopt=AbstractPlus]Angelidis G et al. (2011) Metabolic status regulates ghrelin function on energy homeostasis. Neuroendocrinology93: 48\u201057 [https://www.ncbi.nlm.nih.gov/pubmed/21124019?dopt=AbstractPlus]Briggs DI et al. (2014) Novel and conventional receptors for ghrelin, desacyl\u2010ghrelin, and pharmacologically related compounds. Pharmacol. Rev.66: 984\u20101001 [https://www.ncbi.nlm.nih.gov/pubmed/25107984?dopt=AbstractPlus]Callaghan B et al. (2005) International Union of Pharmacology. LVI. Ghrelin receptor nomenclature, distribution, and function. Pharmacol. Rev.57: 541\u20106 [https://www.ncbi.nlm.nih.gov/pubmed/16382107?dopt=AbstractPlus]Davenport AP NC\u2010IUPHAR Subcommittee on the Glucagon receptor familynomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/12615957?dopt=AbstractPlus]) are activated by the endogenous peptide (27\u201044 aa) hormones http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1136 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5194 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1140 , glucose\u2010dependent insulinotropic polypeptide ), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2270 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3643 . One common precursor (https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:4191) generates http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1136 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5194 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1140 peptides [http://www.ncbi.nlm.nih.gov/pubmed/11179772?dopt=AbstractPlus]. For a recent review on review the current understanding of the structures of GLP\u20101 and GLP\u20101R, the molecular basis of their interaction, and the signaling events associated with it, see de Graaf et al., 2016 [http://www.ncbi.nlm.nih.gov/pubmed/27630114?dopt=AbstractPlus].The glucagon family of receptors , specifically J Diabetes Investig10: 196\u2010201 [https://www.ncbi.nlm.nih.gov/pubmed/30099845?dopt=AbstractPlus]Ahr\u00e9n B. (2019) Glucagon\u2010like peptide\u20101 receptor agonists for type 2 diabetes: A rational drug development. et al. (2018) Glucagon\u2010like peptide 1 in health and disease. Nat Rev Endocrinol14: 390\u2010403 [https://www.ncbi.nlm.nih.gov/pubmed/29728598?dopt=AbstractPlus]Andersen A et al. (2019) Glucagon\u2010like peptide\u20101 receptor agonists in type 2 diabetes treatment: are they all the same? Diabetes Metab. Res. Rev.35: e3070 [https://www.ncbi.nlm.nih.gov/pubmed/30156747?dopt=AbstractPlus]Gentilella R et al. (2016) Glucagon\u2010Like Peptide\u20101 and Its Class B G Protein\u2010Coupled Receptors: A Long March to Therapeutic Successes. Pharmacol. Rev.68: 954\u20101013 [https://www.ncbi.nlm.nih.gov/pubmed/27630114?dopt=AbstractPlus]Graaf Cd et al. (2019) A Review of Practical Issues on the Use of Glucagon\u2010Like Peptide\u20101 Receptor Agonists for the Management of Type 2 Diabetes. Diabetes Ther10: 5\u201019 [https://www.ncbi.nlm.nih.gov/pubmed/30506340?dopt=AbstractPlus]Romera I et al. (2014) GLP\u20101 receptor agonists for type 2 diabetes mellitus: recent developments and emerging agents. Pharmacotherapy34: 1174\u201086 [https://www.ncbi.nlm.nih.gov/pubmed/25382096?dopt=AbstractPlus]Trujillo JM et al. (2017) Cryo\u2010EM structure of the activated GLP\u20101 receptor in complex with a G protein. Nature546: 248\u2010253 [https://www.ncbi.nlm.nih.gov/pubmed/28538729?dopt=AbstractPlus]Zhang Y provisional nomenclature [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) are activated by a non\u2010covalent heterodimeric glycoprotein made up of a common \u03b1\u00a0chainGlycoprotein hormone receptors https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:1885, http://www.uniprot.org/uniprot/P01215), with a unique \u03b2\u00a0chain that confers the biological specificity to http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1157 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1159 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1160 or http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3920 . There is binding cross\u2010reactivity across the endogenous agonists for each of the glycoprotein hormone receptors. The deglycosylated hormones appear to exhibit reduced efficacy at these receptors [http://www.ncbi.nlm.nih.gov/pubmed/2542111?dopt=AbstractPlus]. Structure of follicle\u2010stimulating hormone in complex with the entire ectodomain of its receptor. Proc. Natl. Acad. Sci. U.S.A.109: 12491\u20106 https://www.ncbi.nlm.nih.gov/pubmed/22802634?dopt=AbstractPlusJiang X et al. TSH receptor mutations and disease. http://www.thyroidmanager.org/chapter/tsh\u2010receptor\u2010mutations\u2010and\u2010diseases/. Accessed on 2017\u201002\u201023.Kleinau G et al. (2009) Follicle stimulating hormone receptor mutations and reproductive disorders. Prog Mol Biol Transl Sci89: 115\u201031 https://www.ncbi.nlm.nih.gov/pubmed/20374735?dopt=AbstractPlusTao YX et al. (2013) Structural and functional plasticity of the luteinizing hormone/choriogonadotrophin receptor. Hum. Reprod. Update19: 583\u2010602 https://www.ncbi.nlm.nih.gov/pubmed/23686864?dopt=AbstractPlusTroppmann B 1 and GnRH2 receptors have been cloned from numerous species, most of which express two or three types of GnRH receptor . http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1162 (p\u2010Glu\u2010His\u2010Trp\u2010Ser\u2010Tyr\u2010Gly\u2010LeuArg\u2010Pro\u2010Gly\u2010NH2) is a hypothalamic decapeptide also known as luteinizing hormone\u2010releasing hormone, gonadoliberin, luliberin, gonadorelin or simply as GnRH. It is a member of a family of similar peptides found in many species including http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1164 . Receptors for three forms of GnRH exist in some species but only GnRH I and GnRH II and their cognate receptors have been found in mammals . GnRH1 receptors are expressed by pituitary gonadotrophs, where they mediate the effects of GnRH on gonadotropin hormone synthesis and secretion that underpin central control of mammalian reproduction. GnRH analogues are used in assisted reproduction and to treat steroid hormone\u2010dependent conditions [http://www.ncbi.nlm.nih.gov/pubmed/12072036?dopt=AbstractPlus]. Notably, agonists cause desensitization of GnRH\u2010stimulated gonadotropin secretion and the consequent reduction in circulating sex steroids is exploited to treat hormone\u2010dependent cancers of the breast, ovary and prostate [http://www.ncbi.nlm.nih.gov/pubmed/12072036?dopt=AbstractPlus]. GnRH1 receptors are selectively activated by GnRH I and all lack the COOH\u2010terminal tails found in other GPCRs. GnRH2 receptors do have COOH\u2010terminal tails and (where tested) are selective for GnRH II over GnRH I. GnRH2 receptors are expressed by some primates but not by humans [http://www.ncbi.nlm.nih.gov/pubmed/12538601?dopt=AbstractPlus]. Phylogenetic classifications divide GnRH receptors into three [http://www.ncbi.nlm.nih.gov/pubmed/15082521?dopt=AbstractPlus] or five groups [http://www.ncbi.nlm.nih.gov/pubmed/25344287?dopt=AbstractPlus] and highlight examples of gene loss through evolution, with humans retaining only one ancient gene.GnRH1 and GnRH2 receptors couple primarily to Gq/11 [http://www.ncbi.nlm.nih.gov/pubmed/10734055?dopt=AbstractPlus] but coupling to Gs and Gi is evident in some systems . GnRH2 receptors may also mediate (heterotrimeric) G protein\u2010independent signalling to protein kinases [http://www.ncbi.nlm.nih.gov/pubmed/15059960?dopt=AbstractPlus]. There is increasing evidence for expression of GnRH receptors on hormone\u2010dependent cancer cells where they can exert antiproliferative and/or proapoptotic effects and mediate effects of cytotoxins conjugated to GnRH analogues . In some human cancer cell models http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1164 is more potent than http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1162 , implying mediation by GnRH2 receptors [http://www.ncbi.nlm.nih.gov/pubmed/12237622?dopt=AbstractPlus], but GnRH2 receptors are not expressed by humans because the human GNRHR2 gene contains a frame shift and internal stop codon [http://www.ncbi.nlm.nih.gov/pubmed/12538601?dopt=AbstractPlus]. The possibility remains that this gene generates GnRH2 receptor\u2010related proteins (other than the full\u2010length receptor) that mediate responses to http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1164 (see [http://www.ncbi.nlm.nih.gov/pubmed/11861490?dopt=AbstractPlus]). Alternatively, evidence for multiple active GnRH receptor conformations raises the possibility that GnRH1 receptor\u2010mediated proliferation inhibition in hormone\u2010dependent cancer cells is dependent upon a conformation that couples to Gi rather than Gq/11 proteins as in pituitary cells . Loss\u2010of\u2010function mutations in the GnRH1 receptor and deficiency of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1162 are associated with hypogonadotropic hypogonadism although some \u2019loss of function\u2019 mutations may actually prevent trafficking of \u2019functional\u2019 GnRH1 receptors to the cell surface, as evidenced by recovery of function by nonpeptide antagonists [http://www.ncbi.nlm.nih.gov/pubmed/12843188?dopt=AbstractPlus]. Human GnRH1 receptors are poorly expressed at the cell surface because of failure to meet structural quality control criteria for endoplasmic reticulum exit , and this increases susceptibility to point mutations that further impair trafficking . GnRH receptor signalling may require receptor oligomerisation .GnRHet al. (2017) Expression and Role of Gonadotropin\u2010Releasing Hormone 2 and Its Receptor in Mammals. Front Endocrinol (Lausanne)8: 269 https://www.ncbi.nlm.nih.gov/pubmed/29312140?dopt=AbstractPlusDesaulniers AT et al. (2012) GnRH receptors in cancer: from cell biology to novel targeted therapeutic strategies. Endocr. Rev.33: 784\u2010811 https://www.ncbi.nlm.nih.gov/pubmed/22778172?dopt=AbstractPlusLimonta P In Knobil and Neill's Physiology of Reproduction (4th edition). Edited by Plant TM and Zeleznik AJ.: Elsevier Inc.: [ISBN: 9780123971753]McArdle CA and Roberson MS.. (2015) Gonadotropes and gonadotropin\u2010releasing hormone signaling. et al. (2004) Gonadotropin\u2010releasing hormone receptors. Endocr Rev25: 235\u2010275 https://www.ncbi.nlm.nih.gov/pubmed/15082521?dopt=AbstractPlusMillar RP et al. (2014) Chaperoning G protein\u2010coupled receptors: from cell biology to therapeutics. Endocr. Rev.35: 602\u201047 https://www.ncbi.nlm.nih.gov/pubmed/24661201?dopt=AbstractPlusTao YX provisional nomenclature), although showing little structural similarity to CB1 and CB2 cannabinoid receptors, respond to endogenous agents analogous to the endogenous cannabinoid ligands, as well as some natural/synthetic cannabinoid receptor ligands [http://www.ncbi.nlm.nih.gov/pubmed/21079038?dopt=AbstractPlus]. Although there are multiple reports to indicate that GPR18, GPR55 and GPR119 can be activated in vitro by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3635, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4028 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2661, respectively, there is a lack of evidence for activation by these lipid messengers in vivo. As such, therefore, these receptors retain their orphan status.GPR18, GPR55 and GPR119 International Union of Basic and Clinical Pharmacology. LXXXVIII. G protein\u2010coupled receptor list: recommendations for new pairings with cognate ligands. Pharmacol. Rev.65: 967\u201086 https://www.ncbi.nlm.nih.gov/pubmed/23686350?dopt=AbstractPlusDavenport AP et al. (2016) Biased signaling of lipids and allosteric actions of synthetic molecules for GPR119. Biochem. Pharmacol.119: 66\u201075 https://www.ncbi.nlm.nih.gov/pubmed/27569424?dopt=AbstractPlusHassing HA et al. (2017) Cannabinoid Receptor\u2010Related Orphan G Protein\u2010Coupled Receptors. Adv Pharmacol80: 223\u2010247 https://www.ncbi.nlm.nih.gov/pubmed/28826536Irving A et al. (2015) GPR55: from orphan to metabolic regulator? Pharmacol. Ther.145: 35\u201042 https://www.ncbi.nlm.nih.gov/pubmed/24972076?dopt=AbstractPlusLiu B et al. (2010) International Union of Basic and Clinical Pharmacology. LXXIX. Cannabinoid receptors and their ligands: beyond CB_1 and CB_2. Pharmacol. Rev.62: 588\u2010631 https://www.ncbi.nlm.nih.gov/pubmed/21079038?dopt=AbstractPlusPertwee RG NC\u2010IUPHAR Subcommittee on Histamine Receptorsnomenclature as agreed by the ) are activated by the endogenous ligand http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1204. Marked species differences exist between histamine receptor orthologues [http://www.ncbi.nlm.nih.gov/pubmed/9311023?dopt=AbstractPlus]. The human and rat H3 receptor genes are subject to significant splice variance [http://www.ncbi.nlm.nih.gov/pubmed/16415177?dopt=AbstractPlus]. The potency order of histamine at histamine receptor subtypes is H3 = H4 > H2 > H1 [http://www.ncbi.nlm.nih.gov/pubmed/26084539?dopt=AbstractPlus]. Some agonists at the human H3 receptor display significant ligand bias [http://www.ncbi.nlm.nih.gov/pubmed/27864425?dopt=AbstractPlus]. Antagonists of all 4 histamine receptors have clinical uses: H1 antagonists for allergies (e.g.http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1222), H2 antagonists for acid\u2010reflux diseases (e.g.http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1234), H3 antagonists for narcolepsy and H4 antagonists for atopic dermatitis [http://www.ncbi.nlm.nih.gov/pubmed/26084539?dopt=AbstractPlus] and vestibular neuritis (AUV) , entered and completed vestibular neuritis (AUV) Phase IIa efficacy and safety trials, respectively) .Histamine receptors The histamine autoreceptor is a short isoform of the H_3 receptor. Br. J.Pharmacol.166: 1860\u201071 https://www.ncbi.nlm.nih.gov/pubmed/22356432?dopt=AbstractPlusGbahou F et al. (2016) The Histamine H3 Receptor: Structure, Pharmacology, and Function. Mol. Pharmacol.90: 649\u2010673 https://www.ncbi.nlm.nih.gov/pubmed/27563055?dopt=AbstractPlusNieto\u2010Alamilla G et al. (2015) International Union of Basic and Clinical Pharmacology. XCVIII. Histamine Receptors. Pharmacol. Rev.67: 601\u201055 https://www.ncbi.nlm.nih.gov/pubmed/26084539?dopt=AbstractPlusPanula P et al. (2008) Cloning and characterization of dominant negative splice variants of the human histamine H4 receptor. Biochem. J.414: 121\u201031 [https://www.ncbi.nlm.nih.gov/pubmed/18452403?dopt=AbstractPlus]van Rijn RM http://www.ensembl.org/Homo_sapiens/Gene/Family/Genes?family=ENSFM00500000271913, nomenclature as agreed by theNC\u2010IUPHARSubcommittee on Hydroxycarboxylic acid receptors ) respond to organic acids, including the endogenous hydroxy carboxylic acids 3\u2010hydroxy butyric acid and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2932, as well as the lipid lowering agents http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1588 (niacin), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1596 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1595 . These receptors were provisionally described as nicotinic acid receptors, although nicotinic acid shows submicromolar potency at HCA2 receptors only and is unlikely to be the natural ligand .The hydroxycarboxylic acid family of receptors and https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:4486 (http://www.uniprot.org/uniprot/O00270). Lactate activates HCA1 on adipocytes in an autocrine manner. It inhibits lipolysis and thereby promotes anabolic effects. HCA2 and HCA3 regulate adipocyte lipolysis and immune functions under conditions of increased FFA formation through lipolysis . HCA2 agonists acting mainly through the receptor on immune cells exert antiatherogenic and anti\u2010inflammatory effects. HCA2 is also a receptor for butyrate and mediates some of the beneficial effects of short\u2010chain fatty acids produced by gut microbiota. HCA3 has been shown to be activated by aromatic D\u2010amino acids.Further closely\u2010related GPCRs include the et al. (2008) Nicotinic acid receptor agonists. J. Med. Chem.51: 7653\u201062 https://www.ncbi.nlm.nih.gov/pubmed/18983141?dopt=AbstractPlusBoatman PD et al. (2016) Anti\u2010inflammatory effects of the hydroxycarboxylic acid receptor 2. Metab. Clin. Exp.65: 102\u201013 https://www.ncbi.nlm.nih.gov/pubmed/26773933?dopt=AbstractPlusGraff EC et al. (2013) Recent advances in niacin and lipid metabolism. Curr. Opin. Lipidol.24: 239\u201045 [https://www.ncbi.nlm.nih.gov/pubmed/23619367?dopt=AbstractPlus]Kamanna VS Trends Endocrinol.Metab.28: 227\u2010236 https://www.ncbi.nlm.nih.gov/pubmed/28087125?dopt=AbstractPlusOffermanns S. (2017) Hydroxy\u2010Carboxylic Acid Receptor Actions in Metabolism. et al. (2011) International Union of Basic and Clinical Pharmacology. LXXXII: Nomenclature and Classification of Hydroxy\u2010carboxylic Acid Receptors . Pharmacol. Rev.63: 269\u201090 https://www.ncbi.nlm.nih.gov/pubmed/21454438?dopt=AbstractPlusOffermanns S et al. (2015) Nutritional or pharmacological activation of HCA(2) ameliorates neuroinflammation. Trends Mol Med21: 245\u201055 https://www.ncbi.nlm.nih.gov/pubmed/25766751?dopt=AbstractPlusOffermanns S NC\u2010IUPHAR Subcommittee on the kisspeptin receptornomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/21079036?dopt=AbstractPlus]), like neuropeptide FF (NPFF), prolactin\u2010releasing peptide (PrP) and QRFP receptors responds to endogenous peptides with an arginine\u2010phenylalanine amide (RFamide) motif. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1288 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1284 (KP13) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1283 (https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:6341) (KP10) are biologically\u2010active peptides cleaved from the https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:6341 (http://www.uniprot.org/uniprot/Q15726) gene product. Kisspeptins have roles in, for example, cancer metastasis, fertility/puberty regula tion and glucose homeostasis.The kisspeptin receptor The role of kisspeptin neurons in reproduction and metabolism. J. Endocrinol.238: R173\u2010R183 https://www.ncbi.nlm.nih.gov/pubmed/30042117?dopt=AbstractPlusHarter CJL et al. (2013) Structure, synthesis, and phylogeny of kisspeptin and its receptor. Adv. Exp. Med. Biol.784: 9\u201026 https://www.ncbi.nlm.nih.gov/pubmed/23550000?dopt=AbstractPlusKanda S et al. (2010) International Union of Basic and Clinical Pharmacology. LXXVII. Kisspeptin receptor nomenclature, distribution, and function. Pharmacol. Rev.62: 565\u201078 https://www.ncbi.nlm.nih.gov/pubmed/21079036?dopt=AbstractPlusKirby HR et al. (2009) Kisspeptin signaling in the brain. Endocr. Rev.30: 713\u201043 https://www.ncbi.nlm.nih.gov/pubmed/19770291?dopt=AbstractPlusOakley AE et al. (2014) Molecular evolution of GPCRs: Kisspeptin/kisspeptin receptors. J. Mol. Endocrinol.52: T101\u201017 https://www.ncbi.nlm.nih.gov/pubmed/24577719?dopt=AbstractPlusPasquier J NC\u2010IUPHAR subcommittee on Leukotriene Receptorsnomenclature as agreed by the ) are activated by the endogenous ligands leukotrienes (LT), synthesized from lipoxygenase metabolism of arachidonic acid. The human BLT1 receptor is the high affinity LTB4 receptor whereas the BLT2 receptor in addition to being a low\u2010affinity LTB4 receptor also binds several other lipoxygenase\u2010products, such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3404, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2481, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3401, and the thromboxane synthase product http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6159. The BLT receptors mediate chemotaxis and immunomodulation in several leukocyte populations and are in addition expressed on non\u2010myeloid cells, such as vascular smooth muscle and endothelial cells. In addition to BLT receptors, LTB4 has been reported to bind to the peroxisome proliferator activated receptor (PPAR) \u03b1 [http://www.ncbi.nlm.nih.gov/pubmed/9890897?dopt=AbstractPlus] and the vanilloid TRPV1 ligand\u2010gated nonselective cation channel [http://www.ncbi.nlm.nih.gov/pubmed/16207832?dopt=AbstractPlus]. The receptors for the cysteinyl\u2010leukotrienes aretermed CysLT1 and CysLT2 and exhibit distinct expression patterns in human tissues, mediating for example smooth muscle cell contraction, regulation of vascular permeability, and leukocyte activation. There is also evidence in the literature for additional CysLT receptor subtypes, derived from functional in vitro studies, radioligand binding and in mice lacking both CysLT1 and CysLT2 receptors [http://www.ncbi.nlm.nih.gov/pubmed/24588652?dopt=AbstractPlus]. Cysteinyl\u2010leukotrienes have also been suggested to signal through the P2Y12 receptor , GPR17 [http://www.ncbi.nlm.nih.gov/pubmed/16990797?dopt=AbstractPlus] and GPR99 [http://www.ncbi.nlm.nih.gov/pubmed/23504326?dopt=AbstractPlus].The leukotriene receptors (NC\u2010IUPHAR subcommittee on Leukotriene and Lipoxin Receptorsnomenclatureas agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/24588652?dopt=AbstractPlus]) is activated by the endogenous lipidderived, anti\u2010inflammatory ligands lipoxin A4 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1034) and 15\u2010epiLXA4 . The FPR2/ALX receptor also interacts with endogenous peptide and protein ligands, such as MHC binding peptide [http://www.ncbi.nlm.nih.gov/pubmed/10748237?dopt=AbstractPlus] as well as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1031 (ANXA1) and its N\u2010terminal peptides . In addition, a soluble hydrolytic product of protease action on the urokinase\u2010type plasminogen activator receptor has been reported to activate the FPR2/ALX receptor [http://www.ncbi.nlm.nih.gov/pubmed/11818541?dopt=AbstractPlus]. Furthermore, FPR2/ALX has been suggested to act as a receptor mediating the proinflammatory actions of the acute\u2010phase reactant, serum amyloid A . The agonist activity of the lipid mediators described has been questioned , which may derive from batchto\u2010batch differences, partial agonism or biased agonism. Results from Cooray et al. (2013) [http://www.ncbi.nlm.nih.gov/pubmed/24108355?dopt=AbstractPlus] have addressed this issue and the role of homodimers and heterodimers in intracellular signaling. A receptor selective for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5216 has been suggested from functional studies . Note that the data for FPR2/ALX are also reproduced on the http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=23.The FPR2/ALX receptor (nomenclature agreed by theNC\u2010IUPHAR subcommittee on Oxoeicosanoid Receptors [http://www.ncbi.nlm.nih.gov/pubmed/15001665?dopt=AbstractPlus]) are activated by endogenous chemotactic eicosanoid ligands oxidised at the C\u20105 position, with http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3391 the most potent agonist identified for this receptor. Initial characterization of the heterologously expressed OXE receptor suggested that polyunsaturated fatty acids, such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1051 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3362, acted as receptor antagonists [http://www.ncbi.nlm.nih.gov/pubmed/12065583?dopt=AbstractPlus].Oxoeicosanoid receptors International Union of Pharmacology XLIV. Nomenclature for the Oxoeicosanoid Receptor. Pharmacol. Rev.56: 149\u2010157 https://www.ncbi.nlm.nih.gov/pubmed/15001665?dopt=AbstractPlusBrink C et al. (2003) International Union of Pharmacology XXXVII. Nomenclature for leukotriene and lipoxin receptors. Pharmacol. Rev.55: 195\u2010227 https://www.ncbi.nlm.nih.gov/pubmed/12615958?dopt=AbstractPlusBrink C et al. (2011) International Union of Basic and Clinical Pharmacology. LXXXIV: leukotriene receptor nomenclature, distribution, and pathophysiological functions. Pharmacol. Rev.63: 539\u201084 https://www.ncbi.nlm.nih.gov/pubmed/21771892?dopt=AbstractPlusB\u00e4ck M et al. (2014) Update on leukotriene, lipoxin and oxoeicosanoid receptors: IUPHAR Review 7. Br. J. Pharmacol.171: 3551\u201074 https://www.ncbi.nlm.nih.gov/pubmed/24588652?dopt=AbstractPlusB\u00e4ck M et al. (2012) Cysteinyl leukotriene receptors, old and new; implications for asthma. Clin. Exp. Allergy42: 1313\u201020 https://www.ncbi.nlm.nih.gov/pubmed/22925317?dopt=AbstractPlusLaidlaw TM NC\u2010IUPHAR Subcommittee on Lysophospholipid (LPA) receptors Lysophospholipid Receptorsnomenclature as agreed by the ) are activated by the endogenous phospholipid http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2906. The first receptor, LPA1, was identified as ventricular zone gene\u20101 (vzg\u20101) [http://www.ncbi.nlm.nih.gov/pubmed/8922387?dopt=AbstractPlus], leading to deorphanisation of members of the endothelial differentiation gene (edg) family as other LPA receptors along with sphingosine 1phosphate (S1P) receptors. Additional LPA receptor GPCRs were later identified. Gene names have been codified as LPAR1, etc. to reflect the receptor function of proteins. The crystal structure of LPA1 was solved and demonstrates extracellular LPA access to the binding pocket, consistent with proposed delivery via autotaxin [http://www.ncbi.nlm.nih.gov/pubmed/26091040?dopt=AbstractPlus]. These studies have also implicated cross\u2010talk with endocannabinoids via phosphorylated intermediates that can also activate these receptors. The identified receptors can account for most, although not all, LPA\u2010induced phenomena in the literature, indicating that a majority of LPA\u2010dependent phenomena are receptor\u2010mediated. Binding affinities of unlabeled, natural LPA and AEApto LPA1 weremeasured using backscattering interferometry (pKd = 9) [http://www.ncbi.nlm.nih.gov/pubmed/30463988?dopt=AbstractPlus]. Binding affinities were 77\u2010fold lower than than values obtained using radioactivity [http://www.ncbi.nlm.nih.gov/pubmed/19386608?dopt=AbstractPlus]. Targeted deletion of LPA receptors has clarified signalling pathways and identified physiological and pathophysiological roles. Independent validation by multiple groups has been reported in the peer\u2010reviewed literature for all six LPA receptors described in the tables, including further validation using a distinct read\u2010out via a novel TGF\u03b1 \u201cshedding\u201d assay [http://www.ncbi.nlm.nih.gov/pubmed/22983457?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2906 has also been described as an agonist for the transient receptor potential (Trp) ion channel TRPV1 [http://www.ncbi.nlm.nih.gov/pubmed/22101604?dopt=AbstractPlus] and TRPA1 [http://www.ncbi.nlm.nih.gov/pubmed/28176353?dopt=AbstractPlus]. LPA was originally proposed to be a ligand for GPCR35, but data show that in fact it is a receptor for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6479 [http://www.ncbi.nlm.nih.gov/pubmed/25411203?dopt=AbstractPlus]. All of these proposed non\u2010GPCR receptor identities require confirmation and are not currently recognized as bona fide LPA receptors.Lysophosphatidic acid (LPA) receptors International Union of Basic and Clinical Pharmacology. LXXVIII. Lysophospholipid receptor nomenclature. Pharmacol. Rev.62: 579\u201087 https://www.ncbi.nlm.nih.gov/pubmed/21079037?dopt=AbstractPlusChun J et al. (2014) Lysophospholipid receptor nomenclature review: IUPHAR Review 8. Br. J. Pharmacol.171: 3575\u201094 https://www.ncbi.nlm.nih.gov/pubmed/24602016?dopt=AbstractPlusKihara Y et al. (2015) Lysophosphatidic Acid signaling in the nervous system. Neuron85: 669\u201082 https://www.ncbi.nlm.nih.gov/pubmed/25695267?dopt=AbstractPlusYung YC NC\u2010IUPHAR Subcommittee on Lysophospholipid receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/24602016?dopt=AbstractPlus]) are activated by the endogenous lipid http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=911 (S1P). Originally cloned as orphan members of the endothelial differentiation gene (edg) family, current gene names have been designated as S1P1R through S1P5R [http://www.ncbi.nlm.nih.gov/pubmed/2160972?dopt=AbstractPlus]. S1PRs, particularly S1P1, are expressed throughout all mammalian organ systems. Ligand delivery occurs via two known carriers (or \u201cchaperones\u201d): albumin and HDLbound apolipoprotein M (ApoM), the latter of which elicits biased agonist signaling by S1P1 in multiple cell types . The five S1PRs, two chaperones, and active cellular metabolism have complicated analyses of receptor ligand binding in native systems. Signaling pathways and physiological roles have been characterized through radioligand binding in heterologous expression systems, targeted deletion of the different S1PRs, and most recently, mouse models that report in vivo S1P1R activation . A crystal structure of an S1P1\u2010T4 fusion protein confirmed aspects and binding, specificity, and receptor activation determined previously through biochemical and genetic studies . http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2407 (FTY720), the first drug to target any of the lysophospholipid receptors, binds to four of the five S1PRs, and was the first oral therapy for multiple sclerosis [http://www.ncbi.nlm.nih.gov/pubmed/30625282?dopt=AbstractPlus]. The mechanisms of action of fingolimod and other S1PR modulating drugs in development include binding S1PRs in multiple organ systems, e.g., immune and nervous systems, although the precise nature of their receptor interactions requires clarification .Sphingosine 1\u2010phosphate (S1P) receptors (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2407 (FTY720) is phosphorylated in vivo [http://www.ncbi.nlm.nih.gov/pubmed/16078855?dopt=AbstractPlus] to generatean agonist with activity at S1P1, S1P3, S1P4 and S1P5 receptors . Many of the physiological consequences of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2924 administration, as well as those of other currently described S1P1 agonists, may involve functional antagonism via ubiquitination and subsequent degradation of S1P1 [http://www.ncbi.nlm.nih.gov/pubmed/17237497?dopt=AbstractPlus]. Additionally, receptor specificities of the different compounds may depend on the functional assay system utilized and from which species the receptor sequence originated.The FDA\u2010approved immunomodulator et al. (2016) To fingolimod and beyond: The rich pipeline of drug candidates that target S1P signaling. Pharmacol. Res.113: 521\u2010532 https://www.ncbi.nlm.nih.gov/pubmed/27663260?dopt=AbstractPlusChew WS et al. (2010) International Union of Basic and Clinical Pharmacology. LXXVIII. Lysophospholipid receptor nomenclature. Pharmacol. Rev.62: 579\u201087 https://www.ncbi.nlm.nih.gov/pubmed/21079037?dopt=AbstractPlusChun J et al. (2012) Sphingosine\u20101\u2010phosphate and lymphocyte egress from lymphoid organs. Annu. Rev. Immunol.30: 69\u201094 https://www.ncbi.nlm.nih.gov/pubmed/22149932?dopt=AbstractPlusCyster JG et al. (2017) Sphingosine 1\u2010Phosphate Receptor 1 Signaling in Mammalian Cells. Molecules22: https://www.ncbi.nlm.nih.gov/pubmed/28241498?dopt=AbstractPlusPyne NJ et al. (2013) Sphingosine\u20101\u2010phosphate and its receptors: structure, signaling, and influence. Annu. Rev. Biochem.82: 637\u201062 https://www.ncbi.nlm.nih.gov/pubmed/23527695?dopt=AbstractPlusRosen H et al. (2017) Vascular and Immunobiology of the Circulatory Sphingosine 1\u2010Phosphate Gradient. Annu. Rev. Physiol.79: 67\u201091 https://www.ncbi.nlm.nih.gov/pubmed/27813829?dopt=AbstractPlusYanagida K provisional nomenclature as recommended byNC\u2010IUPHAR [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) are activated by an endogenous nonadecameric cyclic peptide identical in humans and rats generated from a precursor , which also produces http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5374 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5375 .Melanin\u2010concentrating hormone (MCH) receptors Recent updates on the melanin\u2010concentrating hormone (MCH) and its receptor system: lessons from MCH1R antagonists. J. Mol. Neurosci.43: 115\u201021 [https://www.ncbi.nlm.nih.gov/pubmed/20582487?dopt=AbstractPlus]Chung S et al. (2010) Cellular models for the study of the pharmacology and signaling of melaninconcentrating hormone receptors. J. Recept. Signal Transduct. Res.30: 385\u2010402 [https://www.ncbi.nlm.nih.gov/pubmed/21083507?dopt=AbstractPlus]Eberle AN et al. (2005) International Union of Pharmacology. XLVI. G protein\u2010coupled receptor list. Pharmacol Rev57: 279\u2010288 [https://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]Foord SM et al. (2014) Meta\u2010analysis of melanin\u2010concentrating hormone signaling\u2010deficient mice on behavioral and metabolic phenotypes. PLoS ONE9: e99961 [https://www.ncbi.nlm.nih.gov/pubmed/24924345?dopt=AbstractPlus]Takase K provisional nomenclature as recommended byNC\u2010IUPHAR [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) are activated by members of the melanocortin family , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3606\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3606, http://www.uniprot.org/uniprot/P01189) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1333\u2010http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1333, http://www.uniprot.org/uniprot/P01189) forms; \u03b4 form is not found in mammals) and adrenocorticotrophin ). Endogenous antagonists include http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3609 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1335 . ACTH(1\u201024) was approved by the US FDA as a diagnostic agent for adrenal function test, whilst NDP\u2010MSH was approved by EMA for the treatment of erythropoietic protoporphyria. Several synthetic melanocortin receptor agonists are under clinical development.Melanocortin receptors Synaptic changes induced by melanocortin signalling. Nat. Rev. Neurosci.15: 98\u2010110 [https://www.ncbi.nlm.nih.gov/pubmed/24588018?dopt=AbstractPlus]Caruso V et al. (2005) International Union of Pharmacology. XLVI. G protein\u2010coupled receptor list. Pharmacol Rev57: 279\u2010288 [https://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]Foord SM et al. (2011) Physiological roles of the melanocortin MC_3 receptor. Eur. J. Pharmacol.660: 13\u201020 [https://www.ncbi.nlm.nih.gov/pubmed/21211527?dopt=AbstractPlus]Renquist BJ NC\u2010IUPHAR Subcommittee on Melatonin Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/20605968?dopt=AbstractPlus]) are activated by the endogenous ligands http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=224 and clinically used drugs like http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1356, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=198 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7393.Melatonin receptors \u2010AMMTC displays an \u02dc400\u2010fold greater agonist potency than (+)\u2010AMMTC at rat MT1 receptors (see http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3385 for structure) [http://www.ncbi.nlm.nih.gov/pubmed/10433507?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1363 is an MT1/MT2 non\u2010selective competitive melatonin receptor antagonist with about 15\u201025 fold selectivity for the MT2 receptor [http://www.ncbi.nlm.nih.gov/pubmed/9737724?dopt=AbstractPlus]. MT1/MT2 heterodimers present differentpharmacological profiles from MT1 and MT2 receptors [http://www.ncbi.nlm.nih.gov/pubmed/15266022?dopt=AbstractPlus]. The MT3 binding site of hamster brain and peripheral tissues such as kidney and testis, also termed the ML2 receptor, binds selectively http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5396http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5396 [http://www.ncbi.nlm.nih.gov/pubmed/8773460?dopt=AbstractPlus]. Pharmacological investigations of 3MT binding sites have primarily been conducted in hamster tissues. At this site, The endogenous ligand http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5451 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3393 [http://www.ncbi.nlm.nih.gov/pubmed/7798906?dopt=AbstractPlus] appear to function as agonists, while http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=503 [http://www.ncbi.nlm.nih.gov/pubmed/9283717?dopt=AbstractPlus] functions as an antagonist. The 3MT binding site of hamster kidney was also identified as the hamster homologue of human quinone reductase 2 . The 3MT binding site activated by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3393 in eye ciliary body is positively coupled to adenylyl cyclase and regulates chloride secretion [http://www.ncbi.nlm.nih.gov/pubmed/25344385?dopt=AbstractPlus]. Xenopus melanophores and chick brain express a distinct receptor coupled to the Gi/o family of G proteins, for which GPR50 has recently been suggested to be a mammalian counterpart [http://www.ncbi.nlm.nih.gov/pubmed/18400093?dopt=AbstractPlus] although http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=224 does not bind to GPR50 receptors. Several variants of the MTNR1B gene have been associated with increased type 2 diabetes risk [http://www.ncbi.nlm.nih.gov/pubmed/30531911?dopt=AbstractPlus].et al. (2018) Melatonin receptors: molecular pharmacology and signalling in the context of system bias. Br. J. Pharmacol.175: 3263\u20103280 [https://www.ncbi.nlm.nih.gov/pubmed/28707298?dopt=AbstractPlus]Cecon E et al. (2010) International Union of Basic and Clinical Pharmacology. LXXV. Nomenclature, classification, and pharmacology of G protein\u2010coupled melatonin receptors. Pharmacol. Rev.62: 343\u201080 [https://www.ncbi.nlm.nih.gov/pubmed/20605968?dopt=AbstractPlus]Dubocovich ML et al. (2016) Update on melatonin receptors: IUPHAR Review 20. Br. J. Pharmacol.173: 2702\u201025 [https://www.ncbi.nlm.nih.gov/pubmed/27314810?dopt=AbstractPlus]Jockers R et al. (2019) Melatonin in type 2 diabetes mellitus and obesity. Nat Rev Endocrinol15: 105\u2010125 [https://www.ncbi.nlm.nih.gov/pubmed/30531911?dopt=AbstractPlus]Karamitri A et al. (2016) MT1 and MT2 Melatonin Receptors: A Therapeutic Perspective. Annu. Rev. Pharmacol. Toxicol.56: 361\u201083 [https://www.ncbi.nlm.nih.gov/pubmed/26514204?dopt=AbstractPlus]Liu J et al. (2014) MT1 and MT2 melatonin receptors: ligands, models, oligomers, and therapeutic potential. J. Med. Chem.57: 3161\u201085 [https://www.ncbi.nlm.nih.gov/pubmed/24228714?dopt=AbstractPlus]Zlotos DP NC\u2010IUPHAR Subcommittee on Metabotropic Glutamate Receptors[1899]nomenclature as agreed by the ) area family of G protein\u2010coupled receptors activated by the neurotransmitter glutamate. The mGlu family is composed of eight members (named mGlu1 to mGlu8) which are divided in three groups based on similarities of agonist pharmacology, primary sequence and G protein coupling to effector: Group\u2010I (mGlu1 and mGlu5), Group\u2010II (mGlu2 and mGlu3) and Group\u2010III (see Further reading).Metabotropic glutamate (mGlu) receptors domains of both mGlu1 and mGlu5 have been solved, and confirm a general helical organization similar to that of other GPCRs, although the helices appear more compacted . mGlu form constitutive dimers crosslinked by a disulfide bridge. Recent studies revealed the possible formation of heterodimers between either group\u2010I receptors, or within and between group\u2010II and \u2010III receptors [http://www.ncbi.nlm.nih.gov/pubmed/20826542?dopt=AbstractPlus]. Although well characterized in transfected cells, co\u2010localization and specific pharmacological properties also suggest the existence of such heterodimers in the brain .Structurally, mGlu are composed of three juxtaposed domains: a core G protein\u2010activating seven\u2010transmembrane domain (TM), common to all GPCRs, is linked via a rigid cysteine\u2010rich domain (CRD) to the Venus Flytrap domain (VFTD), a large bi\u2010lobed extracellular domain where glutamate binds. The structures of the VFTD of mGluhttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1369, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1411, N\u2010acetylaspartylglutamate (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1405) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5447. Group\u2010I mGlu receptors may be activated by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1367 and (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1366)http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1366 [http://www.ncbi.nlm.nih.gov/pubmed/8532171?dopt=AbstractPlus] and antagonized by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1392 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3350 . Group\u2010III mGlu receptors may be activated by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1410 and (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1406)http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1406 [http://www.ncbi.nlm.nih.gov/pubmed/10336568?dopt=AbstractPlus]. An example of an antagonist selective for mGlu receptors is http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1378, which blocks mGlu2 and mGlu3 at low nanomolar concentrations, mGlu8 at high nanomolar concentrations, and mGlu4, mGlu5, and mGlu7 in the micromolar range [http://www.ncbi.nlm.nih.gov/pubmed/9680254?dopt=AbstractPlus]. In addition to orthosteric ligands that directly interact with the glutamate recognition site, allosteric modulators that bind within the TM domain have been described. Negative allosteric modulators are listed separately. The positive allosteric modulators most often act as \u2018potentiators\u2019 of an orthosteric agonist response, without significantly activating the receptor in the absence of agonist.The endogenous ligands of mGlu are http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1405 as an agonist at mGlu3 receptors was questioned on the basis of contamination with glutamate , but this has been refuted [http://www.ncbi.nlm.nih.gov/pubmed/21740441?dopt=AbstractPlus].The activity of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1391http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1391 [http://www.ncbi.nlm.nih.gov/pubmed/12695537?dopt=AbstractPlus] and [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5392http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5392 [http://www.ncbi.nlm.nih.gov/pubmed/15976016?dopt=AbstractPlus] at mGlu1 receptors and [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1425http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1425 [http://www.ncbi.nlm.nih.gov/pubmed/11814808?dopt=AbstractPlus] and [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5394http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5394 [http://www.ncbi.nlm.nih.gov/pubmed/12438526?dopt=AbstractPlus] at mGlu5 receptors; ). Although a number of radioligands have been used to examine binding in native tissues, correlation with individual subtypes is limited. Many pharmacological agents have not been fully tested across all known subtypes of mGlu receptors and may have unappreciated biased or neutral activity at other subtypes [http://www.ncbi.nlm.nih.gov/pubmed/29514854?dopt=AbstractPlus]. Potential differences linked to the species (e.g. human versus rat or mouse) of the receptors and the receptor splice variants are generally not known. The influence of receptor expression level on pharmacology and selectivity has not been controlled for in most studies, particularly those involving functional assays of receptor coupling.Radioligand binding using a variety of radioligands has been conducted on recombinant receptors at mGlu5 receptors. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1377 also exhibits agonist activity at NMDA glutamate receptors [http://www.ncbi.nlm.nih.gov/pubmed/9106476?dopt=AbstractPlus], and is an antagonist at all Group\u2010III mGluRs with an IC50 of 30\u03bcM. A potential novel metabotropic glutamate receptor coupled to phosphoinositide turnover has been observed in rat brain; it is activated by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5449 (ineffective as an agonist at recombinant Group I metabotropic glutamate receptors), but is resistant to http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1378 [http://www.ncbi.nlm.nih.gov/pubmed/9353394?dopt=AbstractPlus]. There are also reports of a distinct metabotropic glutamatereceptor coupled tophospholipase D in ratbrain, which does not readily fit into the current classification Pharmacology and functions of metabotropic glutamate receptors. Annu. Rev. Pharmacol. Toxicol.37: 205\u2010237 [https://www.ncbi.nlm.nih.gov/pubmed/9131252?dopt=AbstractPlus]Conn PJ et al. (2006) Metabotropic glutamate receptors. Cell Tissue Res.326: 483\u2010504 [https://www.ncbi.nlm.nih.gov/pubmed/16847639?dopt=AbstractPlus]Ferraguti F et al. (2011) Metabotropic glutamate receptors: from the workbench to the bedside. Neuropharmacology60: 1017\u201041 [https://www.ncbi.nlm.nih.gov/pubmed/21036182?dopt=AbstractPlus]Nicoletti F et al. (2010) Metabotropic glutamate receptors: physiology, pharmacology, and disease. Annu. Rev. Pharmacol. Toxicol.50: 295\u2010322 [https://www.ncbi.nlm.nih.gov/pubmed/20055706?dopt=AbstractPlus]Niswender CM et al. (2016) Organization and functions of mGlu and GABAB receptor complexes. Nature540: 60\u201068 [https://www.ncbi.nlm.nih.gov/pubmed/27905440?dopt=AbstractPlus]Pin JP et al. (2011) The complexity of their activation mechanism opens new possibilities for the modulation of mGlu and GABAB class C G protein\u2010coupled receptors. Neuropharmacology60: 82\u201092 [https://www.ncbi.nlm.nih.gov/pubmed/20713070?dopt=AbstractPlus]Rondard P provisional nomenclature) are activated by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1458 , a 22 amino\u2010acid peptide derived from a precursor , which may also generate a http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5376 . These receptors promote gastrointestinal motility and are suggested to be responsible for the gastrointestinal prokinetic effects of certain macrolide antibiotics , although for many of these molecules the evidence is sparse.Motilin receptors are not usually detected in rodents, although brain and other responses to motilin and the macrolide http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1444 have been reported and the mechanism of these actions is obscure . Notably, in some non\u2010laboratory rodents (e.g. the North American kangaroo rat (Dipodomys) and mouse (Microdipodops) a functional form of motilin may exist but the motilin receptor is non\u2010functional [http://www.ncbi.nlm.nih.gov/pubmed/15027861?dopt=AbstractPlus]. Marked differences in ligand affinities for the motilin receptor in dogs and humans may be explained by significant differences in receptor structure [http://www.ncbi.nlm.nih.gov/pubmed/23189978?dopt=AbstractPlus]. Note that for the complex macrolide structures, selectivity of action has often not been rigorously examined and other actions are possible . Small molecule motilin receptor agonists are now described . The motilin receptor does not appear to have constitutive activity [http://www.ncbi.nlm.nih.gov/pubmed/12907757?dopt=AbstractPlus]. Although not proven, the existence of biased agonism at the receptor has been suggested . A truncated 5\u2010transmembrane structure has been identified but this is without activity when transfected into a host cell [http://www.ncbi.nlm.nih.gov/pubmed/10381885?dopt=AbstractPlus]. Receptor dimerisation has not been reported.In terms of structure, the motilin receptor has closest homology with the ghrelin receptor. Thus, the human motilin receptor shares 52% overall amino acid identity with the ghrelin receptor and 86% in the transmembrane regions [et al. (2009) Motilin and ghrelin as prokinetic drug targets. Pharmacol. Ther.123: 207\u201023 [https://www.ncbi.nlm.nih.gov/pubmed/19427331?dopt=AbstractPlus]De Smet B etal. (2018) A randomized, double\u2010blind, placebo\u2010controlled trial of camicinal in Parkinson's disease. Mov. Disord.33: 329\u2010332 [https://www.ncbi.nlm.nih.gov/pubmed/29278279?dopt=AbstractPlus]Marrinan SL et al. (2016) Ghrelin and motilin receptors as drug targets for gastrointestinal disorders. Nat Rev Gastroenterol Hepatol13: 38\u201048 [https://www.ncbi.nlm.nih.gov/pubmed/26392067?dopt=AbstractPlus]Sanger GJ NC\u2010IUPHARprovisional nomenclature as recommended by [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) are activated by the endogenous 25 amino acid peptide neuromedin U , NmU\u201025), a peptide originally isolated from pig spinal cord [http://www.ncbi.nlm.nih.gov/pubmed/3839674?dopt=AbstractPlus]. In humans, NmU\u201025 appears to be the sole product of a precursor gene showing a broad tissue distribution, but which is expressed at highest levels in the upper gastrointestinal tract, CNS, bone marrow and fetal liver. Much shorter versions of NmU are found in some species, but not in human, and are derived at least in some instances from the proteolytic cleavage of the longer NmU. Despite species differences in NmU structure, the C\u2010terminal region is highly conserved and contains biological activity. Neuromedin S ) has also been identified as an endogenous agonist [http://www.ncbi.nlm.nih.gov/pubmed/15635449?dopt=AbstractPlus]. NmS33 is, as its name suggests, a 33 amino\u2010acid product of a precursor protein derived from a single gene and contains an amidated Cterminal heptapeptide identical to NmU. NmS\u201033 appears to activate NMU receptors with equivalent potency to NmU\u201025.Neuromedin U receptors , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3858 or http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3863 [http://www.ncbi.nlm.nih.gov/pubmed/15331768?dopt=AbstractPlus]. A range of radiolabelled ( 125I\u2010), fluorescently labelled and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4787 labelled versions of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1470 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1468 are now commercially available.NMU1 and NMU2 couple predominantly to Get al. (2004) Neuromedin U and its receptors: structure, function, and physiological roles. Pharmacol. Rev.56: 231\u201048 [https://www.ncbi.nlm.nih.gov/pubmed/15169928?dopt=AbstractPlus]Brighton PJ et al. (2009) Neuromedin U: physiology, pharmacology and therapeutic potential. Fundam Clin Pharmacol23: 149\u201057 [https://www.ncbi.nlm.nih.gov/pubmed/19645813?dopt=AbstractPlus]Budhiraja S et al. (2009) Emerging pharmacology and physiology of neuromedin U and the structurally related peptide neuromedin S. Br. J. Pharmacol.158: 87\u2010103 [https://www.ncbi.nlm.nih.gov/pubmed/19519756?dopt=AbstractPlus]Mitchell JD Endocrinology150: 2985\u20107 [https://www.ncbi.nlm.nih.gov/pubmed/19549882?dopt=AbstractPlus]Novak CM. (2009) Neuromedin S and U. provisional nomenclature [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]), which exhibit high affinities for neuropeptide FF and RFamide related peptides . NPFF1 is broadly distributed in the central nervous system with the highest levels found in the limbic system and the hypothalamus. NPFF2 is present in high density in the superficial layers of the mammalian spinal cord where it is involved in nociception and modulation of opioid functions.The Neuropeptide FF receptor family contains two subtypes, NPFF1 and NPFF2 (https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:4523 (http://www.uniprot.org/uniprot/Q9NYM4) shows sequence similarities with NPFF1, NPFF2, PrRP and QRFP receptors. The antagonist http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1486 is selective for NPFF receptors, but does not distinguish between the NPFF1 and NPFF2 subtypes .An orphan receptor et al. (2010) Opioid\u2010modulating properties of the neuropeptide FF system. Biofactors36: 423\u20109 [https://www.ncbi.nlm.nih.gov/pubmed/20803521?dopt=AbstractPlus]Moul\u00e9dous L et al. (2006) Structure\u2010activity relationships of neuropeptide FF and related peptidic and non\u2010peptidic derivatives. Peptides27: 990\u20106 [https://www.ncbi.nlm.nih.gov/pubmed/16490282?dopt=AbstractPlus]Vyas N et al. (2008) Modulatory role of neuropeptide FF system in nociception and opiate analgesia. Neuropeptides42: 1\u201018 [https://www.ncbi.nlm.nih.gov/pubmed/17854890?dopt=AbstractPlus]Yang HY provisional nomenclature [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) responds to the 20 amino\u2010acid peptide neuropeptide S derived from a precursor .The neuropeptide S receptor . The human NPS receptor Asn107Ile displayed similar binding affinity buthigher NPSpotency (by approx. 10\u2010fold) than human NPS receptor Asn107 [http://www.ncbi.nlm.nih.gov/pubmed/16144971?dopt=AbstractPlus]. Several epidemiological studies reported an association between Asn107Ile receptor variant and susceptibility to panic disorders . The SNP Asn107Ile has also been linked to sleep behavior [http://www.ncbi.nlm.nih.gov/pubmed/17903308?dopt=AbstractPlus], inflammatory bowel disease [http://www.ncbi.nlm.nih.gov/pubmed/17854592?dopt=AbstractPlus], schizophrenia [http://www.ncbi.nlm.nih.gov/pubmed/22078257?dopt=AbstractPlus], increased impulsivity and ADHD symptoms [http://www.ncbi.nlm.nih.gov/pubmed/23325374?dopt=AbstractPlus]. Interestingly, a carboxy\u2010terminal splice variant of human NPS receptor was found to be overexpressed in asthmatic patients [http://www.ncbi.nlm.nih.gov/pubmed/15073379?dopt=AbstractPlus].Multiple single\u2010nucleotide polymorphisms (SNP) and several splice variants have been identified in the human NPS receptor. The most interesting of these is an Asn\u2010Ile exchange at position 107 Brain neuropeptide S: via GPCR activation to a powerful neuromodulator of socio\u2010emotional behaviors. Cell Tissue Res.375: 123\u2010132 [https://www.ncbi.nlm.nih.gov/pubmed/30112573?dopt=AbstractPlus]Grund T et al. (2010) Neurobiology, pharmacology, and medicinal chemistry of neuropeptide S and its receptor. Med Res Rev30: 751\u201077 [https://www.ncbi.nlm.nih.gov/pubmed/19824051?dopt=AbstractPlus]Guerrini R et al. (2017) Neuropeptide S receptor ligands: a patent review (2005\u20102016). Expert Opin Ther Pat27: 347\u2010362 [https://www.ncbi.nlm.nih.gov/pubmed/27788040?dopt=AbstractPlus]Ruzza C et al. (2004) Neuropeptide S: a neuropeptide promoting arousal and anxiolytic\u2010like effects. Neuron43: 487\u2010497 [https://www.ncbi.nlm.nih.gov/pubmed/15312648?dopt=AbstractPlus]Xu YL provisional nomenclature [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) is activated by two 23\u2010amino\u2010acid peptides, neuropeptide W ) and neuropeptide B ) . C\u2010terminally extended forms of the peptides and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1502 ) also activate NPBW1[http://www.ncbi.nlm.nih.gov/pubmed/12401809?dopt=AbstractPlus]. Unique to both forms of neuropeptide B is the N\u2010terminal bromination of the first tryptophan residue, and it is from this post\u2010translational modification that the nomenclature NPB is derived. These peptides were first identified from bovine hypothalamus and therefore are classed as neuropeptides. Endogenous variants of the peptides without the N\u2010terminal bromination, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1499 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1500 , were not found to be major components of bovine hypothalamic tissue extracts. The NPBW2 receptor is activated by the short and C\u2010terminal extended forms of neuropeptide W and neuropeptide B [http://www.ncbi.nlm.nih.gov/pubmed/12401809?dopt=AbstractPlus].The neuropeptide BW receptor 1 and 0.98\u201021 nM (NPBW2).\u2010/\u2010 mice show changes in social behavior, suggesting that the NPBW1pathway may have animportant role in theemotional responses of social interaction [http://www.ncbi.nlm.nih.gov/pubmed/21390312?dopt=AbstractPlus].NPBW1http://www.ncbi.nlm.nih.gov/pubmed/26136644?dopt=AbstractPlus]. It has been reported that neuropeptide W may have a key role in the gating of stressful stimuli when mice are exposed to novel environments [http://www.ncbi.nlm.nih.gov/pubmed/27140610?dopt=AbstractPlus].For a review of the contribution of neuropeptide B/W to social dominance, see Watanabe and Yamamoto, 2015 . Computational insights into the binding of antagonists to this receptor have also been described [http://www.ncbi.nlm.nih.gov/pubmed/24938207?dopt=AbstractPlus].Two antagonists have been discovered and reported to have affinity for NPBW1, ML181 and ML250, the latter exhibiting improved selectivity (100 fold) for NPBW1 compared to MCH1 receptors [Front Endocrinol (Lausanne)4: 23 [https://www.ncbi.nlm.nih.gov/pubmed/23515889?dopt=AbstractPlus]Sakurai T. (2013) NPBWR1 and NPBWR2: Implications in Energy Homeostasis, Pain, and Emotion. et al. (2006) Neuropeptide B and W: neurotransmitters in an emerging G\u2010protein\u2010coupled receptor system. Br. J. Pharmacol.148: 1033\u201041 [https://www.ncbi.nlm.nih.gov/pubmed/16847439?dopt=AbstractPlus]Singh G NC\u2010IUPHAR Subcommittee on Neuropeptide Y Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/9549761?dopt=AbstractPlus]) are activated by the endogenous peptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1504 , neuropeptide Y\u2010(3\u201036), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1514 , PYY\u2010(3\u201036) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1512 (PP). The receptor originally identified as the Y3 receptor has been identified as the http://www.guidetopharmacology.org/GRAC/ObjectDisplayForward?objectId=71 . The y6 receptor is a functional gene product in mouse, absent in rat, but contains a frame\u2010shift mutation in primates producing a truncated non\u2010functional gene [http://www.ncbi.nlm.nih.gov/pubmed/8641440?dopt=AbstractPlus]. Many of the agonists exhibit differing degrees of selectivity dependent on the species examined. For example, the potency of PP is greater at the rat Y4 receptor than at the human receptor [http://www.ncbi.nlm.nih.gov/pubmed/9802391?dopt=AbstractPlus]. In addition, many agonists lack selectivity for individual subtypes, but can exhibit comparable potency against pairs of NPY receptor subtypes, or have not been examined for activity at all subtypes. [125I]\u2010PYY or [125I]\u2010NPY can be used to label Y1, Y2, Y5 and y6 subtypes non\u2010selectively, while Bowers ME et al. (1998) XVI. International Union of Pharmacology recommendations for the nomenclature of neuropeptide Y, peptide YY and pancreatic polypeptide receptors. Pharmacol. Rev.50: 143\u2010150 [https://www.ncbi.nlm.nih.gov/pubmed/9549761?dopt=AbstractPlus]Michel MC et al. (2013) Neuropeptide Y receptors: how to get subtype selectivity. Front En docrinol (Lausanne)4: 5 [https://www.ncbi.nlm.nih.gov/pubmed/23382728?dopt=AbstractPlus]Pedragosa\u2010Badia X et al. (2011) The neuropeptide Y system: pathophysiological and therapeutic implications in obesity and cancer. Pharmacol. Ther.131: 91\u2010113 [https://www.ncbi.nlm.nih.gov/pubmed/21439311?dopt=AbstractPlus]Zhang L nomenclature as recommended byNC\u2010IUPHAR [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) are activated by the endogenous tridecapeptide neurotensin (pGlu\u2010Leu\u2010Tyr\u2010Glu\u2010Asn\u2010Lys\u2010Pro\u2010Arg\u2010Arg\u2010Pro\u2010Tyr\u2010Ile\u2010Leu) derived from a precursor , which also generates neuromedin N, an agonist at the NTS2 receptor. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3830) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1574) may be used to label NTS1 and NTS2 receptors at 0.1\u20100.3 and 3\u20105 nM concentrations respectively.Neurotensin receptors appears to be a lowefficacy agonist at the NTS2 receptor [http://www.ncbi.nlm.nih.gov/pubmed/9851594?dopt=AbstractPlus], while the NTS1 receptor antagonist http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1582 is an agonist at NTS2 receptors [http://www.ncbi.nlm.nih.gov/pubmed/9851594?dopt=AbstractPlus]. An additional protein, provisionally termed NTS3 , has been suggested to bind lipoprotein lipase and mediate its degradation [http://www.ncbi.nlm.nih.gov/pubmed/10085125?dopt=AbstractPlus]. It has been reported to interact with the NTS1 receptor [http://www.ncbi.nlm.nih.gov/pubmed/12360476?dopt=AbstractPlus] and the NTS2 receptor [http://www.ncbi.nlm.nih.gov/pubmed/19891061?dopt=AbstractPlus], and has beenimplicated in hormone trafficking and/or neurotensin uptake. A splice variant of the NTS2 receptor bearing 5 transmembrane domains has been identified in mouse [http://www.ncbi.nlm.nih.gov/pubmed/9001400?dopt=AbstractPlus] and later in rat [http://www.ncbi.nlm.nih.gov/pubmed/15637074?dopt=AbstractPlus].et al. (2013) Diverse roles of neurotensin agonists in the central nervous system. Front Endocrinol (Lausanne)4: 36 https://www.ncbi.nlm.nih.gov/pubmed/23526754?dopt=AbstractPlusBoules M et al. (2012) Neurotensin and its receptors in the control of glucose homeostasis. Front Endocrinol (Lausanne)3: 143 https://www.ncbi.nlm.nih.gov/pubmed/23230428?dopt=AbstractPlusMazella J et al. (2009) Cancer, chemistry, and the cell: molecules that interact with the neurotensin receptors. ACS Chem. Biol.4: 503\u201025 https://www.ncbi.nlm.nih.gov/pubmed/19462983?dopt=AbstractPlusMyers RM et al. (2017) Oncogenic role of neurotensin and neurotensin receptors in various cancers. Clin. Exp. Pharmacol. Physiol.44: 841\u2010846 https://www.ncbi.nlm.nih.gov/pubmed/28556374?dopt=AbstractPlusOuyang Q http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1614 (met), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1613 (leu), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1643 (\u03b2\u2010end), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3737 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1620 (dynA), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1622 (dynB), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3669 (Big dyn), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1681 (N/OFQ); http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1623 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3668 are also potential endogenous peptides. The Greek letter nomenclature for the opioid receptors, \u03bc, \u03b4 and \u03ba, is well established, and NC\u2010IUPHAR considers this nomenclature appropriate, along with the symbols spelled out , and the acronyms, MOP, DOP, and KOP. . The human N/OFQ receptor, NOP, is considered \u2019opioid\u2010related\u2019 rather than opioid because, while it exhibits a high degree of structural homology with the conventional opioid receptors [http://www.ncbi.nlm.nih.gov/pubmed/8137918?dopt=AbstractPlus], it displays a distinct pharmacology. Currently there are numerous clinically used drugs, such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1627 and many other opioid analgesics, as well as antagonists such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1638, however only for the \u03bc receptor.Opioid and opioid\u2010like receptors are activated by a variety of endogenous peptides including [\u03bc\u2010receptor in particular may be subject to extensive alternative splicing [http://www.ncbi.nlm.nih.gov/pubmed/24076545?dopt=AbstractPlus], these putative isoforms have not been correlated with any of the subtypesof receptor proposed inyears past. Opioid receptors may heterodimerize with each other or with other 7TM receptors [http://www.ncbi.nlm.nih.gov/pubmed/10385123?dopt=AbstractPlus], and give rise to complexes with a unique pharmacology, however, evidence for such heterodimers in native cells is equivocal and the consequences of this heterodimerization for signalling remains largely unknown. For \u03bc\u2010opioid receptors at least, dimerization does not seem to be required for signalling [http://www.ncbi.nlm.nih.gov/pubmed/19542234?dopt=AbstractPlus]. A distinct met\u2010enkephalin receptor lacking structural resemblance to the opioid receptors listed has been identified and termed an opioid growth factor receptor [http://www.ncbi.nlm.nih.gov/pubmed/11890982?dopt=AbstractPlus].Three naloxone\u2010sensitive opioid receptor genes havebeen identified in humans, and while the http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1623 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3668 have been identified as highly selective, putative endogenous agonists for the \u03bc\u2010opioid receptor. At present, however, the mechanisms for endomorphin synthesis in vivo have not been established, and there is no gene identified that encodes for either. Thus, the status of these peptides as endogenous ligands remains unproven.\u03bc\u2010receptors [http://www.ncbi.nlm.nih.gov/pubmed/19116204?dopt=AbstractPlus] and the identification of biased signalling by opioid receptor ligands, in particular, compounds previously characterized as antagonists [http://www.ncbi.nlm.nih.gov/pubmed/17702750?dopt=AbstractPlus]. Pathway bias for agonists makes general rank orders of potency and efficacy somewhat obsolete, so these do not appear in the table. As ever, the mechanisms underlying the acute and long term regulation of opiod receptor function are the subject of intense investigation and debate.Two areas of increasing importance in defining opioid receptor function are the presence of functionally relevant single nucleotide polymorphisms in human \u03bc and \u03b4 receptors, notably thepositive allosteric modulators and silent allosteric \u201cantagonists\u201d outlined in . Negative allosteric modulation of opioid receptors has been previously suggested [http://www.ncbi.nlm.nih.gov/pubmed/16489449?dopt=AbstractPlus], whether all compounds are acting at a similar site remains to be established.The richness of opioid receptor pharmacology has been enhanced with the recent discovery of allosteric modulators of et al. (2012) \u03ba\u2010opioid receptor/dynorphin system: genetic and pharmacotherapeutic implications for addiction. Trends Neurosci.35: 587\u201096 https://www.ncbi.nlm.nih.gov/pubmed/22709632?dopt=AbstractPlusButelman ER et al. (2015) Challenges for opioid receptor nomenclature: IUPHAR Review 9. Br. J. Pharmacol.172: 317\u201023 https://www.ncbi.nlm.nih.gov/pubmed/24528283?dopt=AbstractPlusCox BM et al. (2011) The delta opioid receptor: an evolving target for the treatment of brain disorders. Trends Pharmacol. Sci.32: 581\u201090 https://www.ncbi.nlm.nih.gov/pubmed/21925742?dopt=AbstractPlusPradhan AA et al. (2013) Regulation of \u03bc\u2010opioid receptors: desensitization, phosphorylation, internalization, and tolerance. Pharmacol. Rev.65: 223\u201054 https://www.ncbi.nlm.nih.gov/pubmed/23321159?dopt=AbstractPlusWilliams JT NC\u2010IUPHAR Subcommittee on Orexin receptors(nomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) are activated by the endogenous polypeptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1697 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1699 derived from a common precursor, https://www.genenames.org/data/gene\u2010symbol\u2010report/%23!/hgnc_id/HGNC:4847, by proteolytic cleavage and some typical peptide modifications [http://www.ncbi.nlm.nih.gov/pubmed/9491897?dopt=AbstractPlus]. Currently the only orexin receptor ligand in clinical use is http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2890,whichisused as a hypnotic. Orexin receptor crystal structures have been solved .Orexin receptors q/11 proteins is rather speculative and based on the strong activation of phospholipase C, though recent studies in recombinant cells also stress the importance of Gq/11 [http://www.ncbi.nlm.nih.gov/pubmed/27237973?dopt=AbstractPlus]. Coupling of both receptors to Gi/o and Gs has also been reported . For most native cellular responses observed, the G protein pathway is unknown. The relative potency order of endogenous ligands depends on the cellular signal transduction machinery [http://www.ncbi.nlm.nih.gov/pubmed/23034387?dopt=AbstractPlus]. Similarly, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1700, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=9305 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=10277 may show variable selectivity for OX2 receptors and are also likely to activate OX1 receptors . Many antagonists and radioligands are not well\u2010characterized, and thus the affinities are uncertain. Among radioligands, \u2010TCS 1102 [http://www.ncbi.nlm.nih.gov/pubmed/24376396?dopt=AbstractPlus] and Rhodamine Green\u2010orexinA [http://www.ncbi.nlm.nih.gov/pubmed/11266181?dopt=AbstractPlus] are also useful radioligand tools. Orexin receptors have been reported to be able to form complexes with each other and some other GPCRs as well as \u03c31 receptors, which might affect the signaling and pharmacology . Loss\u2010of\u2010function mutations in the gene encoding the OX2 receptor underlie canine hereditary narcolepsy [http://www.ncbi.nlm.nih.gov/pubmed/10458611?dopt=AbstractPlus]. Antagonists of the orexin receptors are the focus of major drug discovery efforts for their potential to treat insomnia and other disorders of wakefulness [http://www.ncbi.nlm.nih.gov/pubmed/26317591?dopt=AbstractPlus], while agonists would likely be useful in human narcolepsy.The primary coupling of orexin receptors to Get al. (2015) Orexin/hypocretin role in reward: implications for opioid and other addictions. Br. J. Pharmacol.172: 334\u201048 [https://www.ncbi.nlm.nih.gov/pubmed/24641197?dopt=AbstractPlus]Baimel C Neuropharmacology [https://www.ncbi.nlm.nih.gov/pubmed/30347195?dopt=AbstractPlus]Burdakov D. (2018) Reactive and predictive homeostasis: Roles of orexin/hypocretin neurons. Am. J. Physiol., Cell Physiol.304: C2\u201032 [https://www.ncbi.nlm.nih.gov/pubmed/23034387?dopt=AbstractPlus]Kukkonen JP. (2013) Physiology of the orexinergic/hypocretinergic system: a revisit in 2012. et al. (2016) Hypocretins, Neural Systems, Physiology, and Psychiatric Disorders. Curr Psychiatry Rep18: 7 [https://www.ncbi.nlm.nih.gov/pubmed/26733323?dopt=AbstractPlus]Li SB et al. (2014) Motivational activation: a unifying hypothesis of orexin/hypocretin function. Nat. Neurosci.17: 1298\u2010303 [https://www.ncbi.nlm.nih.gov/pubmed/25254979?dopt=AbstractPlus]Mahler SV NC\u2010IUPHARNomenclature as recommended by [http://www.ncbi.nlm.nih.gov/pubmed/23686350?dopt=AbstractPlus].et al. (2013) International Union of Basic and Clinical Pharmacology. LXXXVIII. G protein\u2010coupled receptor list: recommendations for new pairings with cognate ligands. Pharmacol. Rev.65: 967\u201086 [https://www.ncbi.nlm.nih.gov/pubmed/23686350?dopt=AbstractPlus]Davenport AP Oxgr1. Curr. Opin. Nephrol. Hypertens.26: 426\u2010433 [https://www.ncbi.nlm.nih.gov/pubmed/28771454]Grimm PR and Welling PA (2017) alpha\u2010Ketoglutarate drives electroneutral NaCl reabsorption in intercalated cells by activating a G\u2010protein coupled receptor. NC\u2010IUPHAR Subcommittee on P2Y Receptorsnomenclature as agreed by the ) are activated by the endogenous ligands http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1713, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1712, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1734, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1749 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1783. The relationship of many of the cloned receptors to endogenously expressed receptors is not yet established and so it might be appropriate to use wording such as \u2019http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1734\u2010preferring P2Y receptor\u2019 or \u2019P2Y1\u2010like\u2019, etc., until further, as yet undefined, corroborative criteria can be applied . Clinically used drugs acting on these receptors include the dinucleoside polyphosphate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1736, agonist of the P2Y2 receptor subtype, approved in Japan for the management of dry eye disease [http://www.ncbi.nlm.nih.gov/pubmed/24511227?dopt=AbstractPlus], and the P2Y12 receptor antagonists http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7562, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1765 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1776, all approved as antiplatelet drugs .P2Y receptors was also shown to be a biased agonist at P2Y11, this is still under debate . A group of single nucleotide polymorphisms in the P2Y12 gene, forming the so called P2Y12 H2 haplotype, has been associated with increased platelet responsiveness to ADP, increased risk of peripheral arterial disease and with coronary artery disease [http://www.ncbi.nlm.nih.gov/pubmed/17803810?dopt=AbstractPlus]. The platelet\u2010type bleeding disorder due to P2Y12 receptor defects is an autosomal recessive condition characterized by mild to moderate mucocutaneous bleeding and excessive bleeding after surgery or trauma. The defect is due to the inability of ADP to induce platelet aggregation [http://www.ncbi.nlm.nih.gov/pubmed/12578987?dopt=AbstractPlus]. The P2Y13 receptor Met\u2010158\u2010Thr polymorphism, which is in linkage disequilibrium with the P2Y12 locus, is not associated with acute myocardial infarction, diabetes mellitus or related risk factors [http://www.ncbi.nlm.nih.gov/pubmed/18213371?dopt=AbstractPlus]. The P2Y14 receptor was previously considered to exclusively bind sugar nucleotides such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1783 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1782 [http://www.ncbi.nlm.nih.gov/pubmed/10753868?dopt=AbstractPlus]. However, more recent evidence with several cell lines has demonstrated that http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1749 (UDP) is 5\u2010fold more potent than http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1783 [http://www.ncbi.nlm.nih.gov/pubmed/19759354?dopt=AbstractPlus]. UDP was also shown to competitively antagonise the UDP\u2010glucose response at the human recombinant P2Y14 receptor [http://www.ncbi.nlm.nih.gov/pubmed/18252808?dopt=AbstractPlus].Single nucleotide polymorphisms of the P2YRet al. (2006) International Union of Pharmacology LVIII: update on the P2Y G protein\u2010coupled nucleotide receptors: frommolecular mechanisms andpathophysiology totherapy. Pharmacol. Rev.58: 281\u2010341 [https://www.ncbi.nlm.nih.gov/pubmed/16968944?dopt=AbstractPlus]Abbracchio MP et al. (2015) Nucleotides Acting at P2Y Receptors: Connecting Structure and Function. Mol. Pharmacol.88: 220\u201030 [https://www.ncbi.nlm.nih.gov/pubmed/25837834?dopt=AbstractPlus]Jacobson KA et al. (2016) Pharmacology and structure of P2Y receptors. Neuropharmacology104: 50\u201061 [https://www.ncbi.nlm.nih.gov/pubmed/26519900?dopt=AbstractPlus]von K\u00fcgelgen I NC\u2010IUPHAR Subcommittee on Parathyroid Hormone Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/25713287?dopt=AbstractPlus]) are family B G protein\u2010coupled receptors. The parathyroid hormone (PTH)/parathyroid hormone\u2010related peptide (PTHrP) receptor (PTH1 receptor) is activated by precursor\u2010derived peptides: http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1785 (84 amino acids), and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3738 (141 amino\u2010acids) and related peptides (PTH\u2010(1\u201034), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1790, http://www.uniprot.org/uniprot/P12272)). The parathyroid hormone 2 receptor (PTH2 receptor) is activated by the precursor\u2010derived peptide http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1815 (39 amino acids). [125I]PTH may be used to label both PTH1 and PTH2 receptors.The parathyroid hormone receptors (The parathyroid hormone type 1 receptor (PTHR) is the canonical GPCR for PTH and PTHrP. It is coupled to Gs and Gq and regulates the development of bone, heart, mammary glands and other tissues in response to PTHrP, and blood concentrations of calcium and phosphate ions, as well as vitamin D, in response to PTH. Another important action of the PTH/PTHR system is to stimulate bone formation when the hormone is intermittently administrated (daily injection).http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1785 is an agonist at human PTH2 receptors, it fails to activate the rodent orthologues. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1815 is a weak antagonist at PTH1 receptors [http://www.ncbi.nlm.nih.gov/pubmed/11159842?dopt=AbstractPlus].Although et al. (2015) PTH receptor\u20101 signalling\u2010mechanistic insights and therapeutic prospects. Nat Rev Endocrinol11: 712\u201024 [https://www.ncbi.nlm.nih.gov/pubmed/26303600?dopt=AbstractPlus]Cheloha RW et al. (2015) International Union of Basic and Clinical Pharmacology. XCIII. The Parathyroid Hormone Receptors\u2010Family B G Protein\u2010Coupled Receptors. Pharmacol. Rev.67: 310\u201037 [https://www.ncbi.nlm.nih.gov/pubmed/25713287?dopt=AbstractPlus]Gardella TJ et al. (2014) Endosomal generation of cAMP in GPCR signaling. Nat. Chem. Biol.10: 700\u20106 [https://www.ncbi.nlm.nih.gov/pubmed/25271346?dopt=AbstractPlus]Vilardaga JP http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1831, 1\u2010O\u2010alkyl\u20102\u2010acetyl\u2010sn\u2010glycero\u20103\u2010phosphocholine) is an ether phospholipid mediator associated with platelet coagulation, but also subserves inflammatory roles. The PAF receptor is activated by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1831 and other suggested endogenous ligands are oxidized phosphatidylcholine [http://www.ncbi.nlm.nih.gov/pubmed/10497200?dopt=AbstractPlus] and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2508 [http://www.ncbi.nlm.nih.gov/pubmed/9038918?dopt=AbstractPlus]. It may also be activated by bacterial lipopolysaccharide [http://www.ncbi.nlm.nih.gov/pubmed/1333988?dopt=AbstractPlus].Platelet\u2010activating factor (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1859 Kd 44.6 nM) is currently unavailable.Note that a previously recommended radioligand International Union of Pharmacology. XLVI. G protein\u2010coupled receptor list. Pharmacol Rev57: 279\u2010288 [https://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]Foord SM et al. (2000) Platelet\u2010activating factor (PAF) receptor and genetically engineered PAF receptor mutant mice. Prog. Lipid Res.39: 41\u201082 [https://www.ncbi.nlm.nih.gov/pubmed/10729607?dopt=AbstractPlus]Ishii S et al. (2000) Platelet\u2010activating factor and related lipid mediators. Annu. Rev. Biochem.69: 419\u201045 [https://www.ncbi.nlm.nih.gov/pubmed/10966465?dopt=AbstractPlus]Prescott SM 1 and PKR2 respond to the cysteine\u2010rich 81\u201086 amino\u2010acid peptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1866 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1867 (protein Bv8 homologue). An orthologue of PROK1 from black mamba (Dendroaspis polylepis) venom, mamba intestinal toxin 1 is a potent, nonselective agonist at prokineticin receptors [http://www.ncbi.nlm.nih.gov/pubmed/12054613?dopt=AbstractPlus], while http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5362, an orthologue of PROK2 from amphibians , is equipotent at recombinant PKR1 and PKR2 [http://www.ncbi.nlm.nih.gov/pubmed/16113687?dopt=AbstractPlus], and has high potency in macrophagechemotaxis assays, which arelost in PKR1null mice.Prokineticin receptors, PKRPROKR1 are associated with Hirschsprung's disease [http://www.ncbi.nlm.nih.gov/pubmed/21858136?dopt=AbstractPlus], while genetic mutations in PROKR2 are associated with hypogonadotropic hypogonadism with anosmia [http://www.ncbi.nlm.nih.gov/pubmed/23596439?dopt=AbstractPlus], hypopituitarism with pituitary stalk interruption [http://www.ncbi.nlm.nih.gov/pubmed/22466334?dopt=AbstractPlus] and Hirschsprung's disease [http://www.ncbi.nlm.nih.gov/pubmed/21858136?dopt=AbstractPlus]. PKR2 has been recently identified as a receptor for T. cruzi natural infection [http://www.ncbi.nlm.nih.gov/pubmed/25324134?dopt=AbstractPlus].Genetic mutations in et al. (2011) Prokineticin receptor 1 (PKR1) signalling in cardiovascular and kidney functions. Cardiovasc. Res.92: 191\u20108 [https://www.ncbi.nlm.nih.gov/pubmed/21856786?dopt=AbstractPlus]Boulberdaa M et al. (2018) The Prokineticins: Neuromodulators and Mediators of Inflammation and Myeloid Cell\u2010Dependent Angiogenesis. Physiol. Rev.98: 1055\u20101082 [https://www.ncbi.nlm.nih.gov/pubmed/29537336?dopt=AbstractPlus]Negri L et al. (2012) Bv8/PK2 and prokineticin receptors: a druggable pronociceptive system. Curr Opin Pharmacol12: 62\u20106 [https://www.ncbi.nlm.nih.gov/pubmed/22136937?dopt=AbstractPlus]Negri L et al. (2007) Bv8/Prokineticin proteins and their receptors. Life Sci.81: 1103\u201016 [https://www.ncbi.nlm.nih.gov/pubmed/17881008?dopt=AbstractPlus]Negri L et al. (2008) Prokineticin\u2010signaling pathway. Int. J. Biochem. Cell Biol.40: 1679\u201084 [https://www.ncbi.nlm.nih.gov/pubmed/18440852?dopt=AbstractPlus]Ngan ES https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:17945, http://www.uniprot.org/uniprot/P81277) for PrRP generates 31 and 20\u2010amino\u2010acid versions. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3665 is a 43 amino acid peptide derived from https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:29982 (http://www.uniprot.org/uniprot/P83859) and is also known as P518 or 26RFa. RFRP is an RF amide\u2010related peptide [http://www.ncbi.nlm.nih.gov/pubmed/11025660?dopt=AbstractPlus] derived from a FMRFamide\u2010related peptide precursor , which is cleaved to generate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3736 , neuropeptide http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5340 , neuropeptide http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5373 and neuropeptide http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4016 (neuropeptide NPVF).The precursor (https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:4523 (http://www.uniprot.org/uniprot/Q9NYM4) shows sequence similarities with NPFF1, NPFF2, PrRP and QRFP receptors.The orphan receptor et al. (2006) Prolactin releasing peptide (PrRP): an endogenous regulator of cell growth. Peptides27: 1099\u2010103 [https://www.ncbi.nlm.nih.gov/pubmed/16500730?dopt=AbstractPlus]Samson WK et al. (2010) Roles of prolactin\u2010releasing peptide and RFamide related peptides in the control of stress and food intake. FEBS J.277: 4998\u20105005 [https://www.ncbi.nlm.nih.gov/pubmed/21126313?dopt=AbstractPlus]Takayanagi Y NC\u2010IUPHAR Subcommittee on Prostanoid Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/21752876?dopt=AbstractPlus]) are activated by the endogenous ligands prostaglandins http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1881, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1882, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1883, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1884, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4483, prostacyclin [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1915] and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4482. Measurement of the potency of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1915 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4482 is hampered by their instability in physiological salt solution; they are often replaced by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1917 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1888, respectively, in receptor characterization studies.Prostanoid receptors coupling . The number of EP3 receptor (protein) variants are variable depending on species, with five in human, three in rat and three in mouse. Putative receptor(s) for prostamide F (which as yet lack molecular correlates) and which preferentially recognize http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5456 and its analogues (e.g. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1958) have been identified, together with moderate\u2010potency antagonists (e.g.http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5455) [http://www.ncbi.nlm.nih.gov/pubmed/18700152?dopt=AbstractPlus].The EPhttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3386, used in in vitro studies, has a EC50 of 15nM which is the concentration of the compound giving half\u2010maximal stimulation of inositol phosphate turnover in HEK\u2010293 cells expressing the human FP receptor [http://www.ncbi.nlm.nih.gov/pubmed/17076623?dopt=AbstractPlus].The free acid form of AL\u201012182, http://www.ncbi.nlm.nih.gov/pubmed/1830308?dopt=AbstractPlus] and STA2 [http://www.ncbi.nlm.nih.gov/pubmed/8242228?dopt=AbstractPlus] use human platelets as the source of TP receptors for competition radio\u2010ligand binding assays to determine the indicated activity values.References given alongside the TP receptor agonists I\u2010BOP and in the BEAS\u20102B human airway epithelial cell line [http://www.ncbi.nlm.nih.gov/pubmed/21173040?dopt=AbstractPlus] is available. This receptor is selectively activated by 15R\u201017,18,19,20\u2010tetranor\u201016\u2010m\u2010tolylisocarbacyclin (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5864) and 15R\u2010Deoxy 17,18,19,20\u2010tetranor\u201016m\u2010tolyl\u2010isocarbacyclin (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5865). However, molecular biological evidence for an IP2 subtype is currently lacking.Pharmacological evidence for a second IP receptor, denoted IPet al. (2011) International union of basic and clinical pharmacology. LXXXIII: classification of prostanoid receptors, updating 15 years of progress. Pharmacol. Rev.63: 471\u2010538 [https://www.ncbi.nlm.nih.gov/pubmed/21752876?dopt=AbstractPlus]Woodward DF NC\u2010IUPHAR Subcommittee on Proteinase\u2010activated Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/12037136?dopt=AbstractPlus]) are unique members of the GPCR superfamily activated by proteolytic cleavage of their amino terminal exodomains. Agonist proteinase\u2010induced hydrolysis unmasks a tethered ligand (TL) at the exposed amino terminus, which acts intramolecularly at the binding site in the body of the receptor to effect transmembrane signalling. TL sequences at human PAR1\u20104 are http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5361, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3740, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5360 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3739, respectively. With the exception of PAR3, synthetic peptides with these sequences are able to act as agonists at their respective receptors. Several proteinases, including neutrophil elastase, cathepsin G and chymotrypsin can have inhibitory effects at PAR1 and PAR2 such that they cleave the exodomain of the receptor without inducing activation of G\u03b1q\u2010coupled calcium signalling, thereby preventing activation by activating proteinases but not by agonist peptides. Neutrophil elastase (NE) cleavage of PAR1 and PAR2 can however activate MAP kinase signaling by exposing a TL that is different from the one revealed by trypsin [http://www.ncbi.nlm.nih.gov/pubmed/22212680?dopt=AbstractPlus]. PAR2 ectivation by NE regulates inflammation and pain responses and triggers mucin secretion from airway epithelial cells [http://www.ncbi.nlm.nih.gov/pubmed/23392769?dopt=AbstractPlus].Proteinase\u2010activated receptors , generated by the action of Factor X on liver\u2010derived prothrombin ; trypsin, generated by the action of enterokinase on pancreatic\u2010derived trypsinogen ; tryptase, a family of enzymes secreted from mast cells; cathepsin G generated from leukocytes; liver\u2010derived protein C generated in plasma by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4453 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6655 .Endogenous serine proteases (EC 3.4.21.) active at the proteinase\u2010activated receptors include: et al. (2011) Structure, function and pathophysiology of protease activated receptors. Pharmacol. Ther.130: 248\u201082 https://www.ncbi.nlm.nih.gov/pubmed/21277892?dopt=AbstractPlusAdams MN et al. (2012) Allosteric modulation of protease\u2010activated receptor signaling. Mini Rev Med Chem12: 804\u201011 https://www.ncbi.nlm.nih.gov/pubmed/22681248?dopt=AbstractPlusCanto I et al. (2010) The role of thrombin and protease\u2010activated receptors in pain mechanisms. Thromb. Haemost.103: 1145\u201051 https://www.ncbi.nlm.nih.gov/pubmed/20431855?dopt=AbstractPlusGarc\u00eda PS et al. (2002) International Union of Pharmacology. XXVIII. Proteinase\u2010activated receptors. Pharmacol. Rev.54: 203\u201017 https://www.ncbi.nlm.nih.gov/pubmed/12037136?dopt=AbstractPlusHollenberg MD et al. (2012) Targeting proteinase\u2010activated receptors: therapeutic potential and challenges. Nat Rev Drug Discov11: 69\u201086 https://www.ncbi.nlm.nih.gov/pubmed/22212680?dopt=AbstractPlusRamachandran R et al. (2010) Signal transduction by protease\u2010activated receptors. Br. J. Pharmacol.160: 191\u2010203 https://www.ncbi.nlm.nih.gov/pubmed/20423334?dopt=AbstractPlusSoh UJ NC\u2010IUPHAR Subcommittee on the QRFP receptornomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/28613414?dopt=AbstractPlus]; QRFPR, formerly known as the Peptide P518 receptor), previously designated as an orphan GPCR receptor was identified in 2001 by Lee et al. from a hypothalamus cDNA library [http://www.ncbi.nlm.nih.gov/pubmed/11574155?dopt=AbstractPlus]. However, the reported cDNA (AF411117) is a chimera with bases 1\u2010127 derived from chromosome 1 and bases 155\u20101368 derived from chromosome 4. When corrected, QRFPR encodes a 431 amino acid protein that shares sequence similarities in the transmembrane spanning regions with other peptide receptors. These include neuropeptide FF2 (38%), neuropeptide Y2 (37%) and galanin Gal1 (35%) receptors.The human gene encoding the QRFP receptor (http://www.guidetopharmacology.org/GRAC/ (http://www.uniprot.org/uniprot/9NYM4) shows sequence similarities with the QRFP receptor, as well as with the NPFF1, NPFF2, and PrRP receptors.The orphan receptor et al. (2011) The RFamide neuropeptide 26RFa and its role in the control of neuroendocrine functions. Front Neuroendocrinol32: 387\u201097 https://www.ncbi.nlm.nih.gov/pubmed/21530572?dopt=AbstractPlusChartrel N et al. (2006) Recent advances in mammalian RFamide peptides: the discovery and functional analyses of PrRP, RFRPs and QRFP. Peptides27: 1073\u201086 https://www.ncbi.nlm.nih.gov/pubmed/16500002?dopt=AbstractPlusFukusumi S et al. (2017) The Arg\u2010Phe\u2010amide peptide 26RFa/glutamine RF\u2010amide peptide and its receptor: IUPHAR Review 24. Br. J. Pharmacol.174: 3573\u20103607 https://www.ncbi.nlm.nih.gov/pubmed/28613414?dopt=AbstractPlusLeprince J NC\u2010IUPHAR Subcommittee on Relaxin family peptide receptorsnomenclature as agreed by the ) may be divided into two pairs, RXFP1/2 and RXFP3/4. Endogenous agonists at these receptors are heterodimeric peptide hormones structurally related to insulin: http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1988 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1989 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1990 , insulin\u2010like peptide 3 ) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2000 . Species homologues of relaxin have distinct pharmacology and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1989 interacts with RXFP1, RXFP2 and RXFP3, whereas mouse and rat relaxin selectively bind to and activate RXFP1 [http://www.ncbi.nlm.nih.gov/pubmed/15956680?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1990 is the ligand for RXFP3 but it also binds to RXFP1 and RXFP4 and has differential affinity for RXFP2 between species [http://www.ncbi.nlm.nih.gov/pubmed/15956681?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2000 is the ligand for RXFP4 but is a weak antagonist of RXFP3. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1989 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1995 have multiple complex binding interactions with RXFP1 [http://www.ncbi.nlm.nih.gov/pubmed/27088579?dopt=AbstractPlus] and RXFP2 [http://www.ncbi.nlm.nih.gov/pubmed/30594862?dopt=AbstractPlus] which direct the N\u2010terminal LDLa modules of the receptors together with a linker domain to act as a tethered ligand to direct receptor signaling [http://www.ncbi.nlm.nih.gov/pubmed/16963451?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2000 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1990 interact with their receptors using distinct residues in their B\u2010chains for binding, and activation, respectively .Relaxin family peptide receptors is the cognate peptide ligand for RXFP1 and is in extended Phase III clinical trials for the treatment of acute heart failure [http://www.ncbi.nlm.nih.gov/pubmed/23273292?dopt=AbstractPlus]. Relaxin has vasodilatory, anti\u2010fibrotic, angiogenic, anti\u2010apoptotic and anti\u2010inflammatory effects. A small molecule allosteric agonists http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=8322 has been developed , and a relaxin B\u2010chain mimetic peptide B7\u201033 has been developed which that has cell specific signaling properties [http://www.ncbi.nlm.nih.gov/pubmed/30155023?dopt=AbstractPlus]. The antifibrotic actions of relaxin are dependent on the angiotensin receptor AT2 [http://www.ncbi.nlm.nih.gov/pubmed/24429402?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1995 is the cognate peptide for RXFP2 and is a circulating hormone that in males is essential for testicular descent in utero [http://www.ncbi.nlm.nih.gov/pubmed/10391220?dopt=AbstractPlus] and in females has important roles in ovarian follicle function [http://www.ncbi.nlm.nih.gov/pubmed/30204868?dopt=AbstractPlus]. In adults, INSL3 has potential roles in testicular function [http://www.ncbi.nlm.nih.gov/pubmed/20952422?dopt=AbstractPlus] and the musculoskeletal system [http://www.ncbi.nlm.nih.gov/pubmed/30625346?dopt=AbstractPlus]. RXFP2 is also present in brain, associated with cortico\u2010thalamic motor circuits [http://www.ncbi.nlm.nih.gov/pubmed/18706979?dopt=AbstractPlus]. cAMP elevation is the major signalling pathway for both RXFP1 and RXFP2 , but RXFP1 also activates MAP kinases, nitric oxide signalling, and tyrosine kinase phosphorylation; and relaxin can interact with glucocorticoid receptors [http://www.ncbi.nlm.nih.gov/pubmed/17293890?dopt=AbstractPlus]. Receptor expression profiles suggest that RXFP3 is a brain neuropeptide receptor and RXFP4 a gut hormone receptor. The brain relaxin\u20103/RXFP3 system modulates feeding via effects in hypothalamus , anxiety , reward and motivated, goal\u2010directed behaviours , and spatial and social memory . Of the other relaxin peptides, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1990 is anagonist atRXFP3 and RXFP4 whereashttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2000 is an agonist at RXFP4 and a weak antagonist at RXFP3. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2000 is secreted from enteroendocrine L cells and the INSL5/RXFP4 system affects food intake [http://www.ncbi.nlm.nih.gov/pubmed/25028498?dopt=AbstractPlus] and glucose homeostasis [http://www.ncbi.nlm.nih.gov/pubmed/25514935?dopt=AbstractPlus]. RXFP3 and RXFP4 couple to Gi/o and inhibit adenylyl cyclase , and also cause Erk1/2 phosphorylation [http://www.ncbi.nlm.nih.gov/pubmed/20159943?dopt=AbstractPlus]. RXFP4 also causes phosphorylation of p38MAPK, Akt and S6RP [http://www.ncbi.nlm.nih.gov/pubmed/27243554?dopt=AbstractPlus] and GLP\u20101 secretion in vitro [http://www.ncbi.nlm.nih.gov/pubmed/29535183?dopt=AbstractPlus]. There is evidence that at RXFP3, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1989 is a biased ligand compared to the cognate ligand http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1990 [http://www.ncbi.nlm.nih.gov/pubmed/20159943?dopt=AbstractPlus].et al. (2013) Relaxin family peptides and their receptors. Physiol. Rev.93: 405\u201080 https://www.ncbi.nlm.nih.gov/pubmed/23303914?dopt=AbstractPlusBathgate RA et al. (2010) Cardiovascular effects of relaxin: from basic science to clinical therapy. Nat Rev Cardiol7: 48\u201058 https://www.ncbi.nlm.nih.gov/pubmed/19935741?dopt=AbstractPlusDu XJ et al. (2015) International Union of Basic and Clinical Pharmacology. XCV. Recent advances in the understanding of the pharmacology and biological roles of relaxin family peptide receptors 1\u20104, the receptors for relaxin family peptides. Pharmacol. Rev.67: 389\u2010440 https://www.ncbi.nlm.nih.gov/pubmed/25761609?dopt=AbstractPlusHalls ML etal. (2011) Relaxin family peptides in the male reproductive system\u2010a critical appraisal. Mol. Hum. Reprod.17: 71\u201084 https://www.ncbi.nlm.nih.gov/pubmed/20952422?dopt=AbstractPlusIvell R 1\u2010SST5; NC\u2010IUPHAR Subcommittee on Somatostatin Receptorsnomenclature as agreed by the [http://www.ncbi.nlm.nih.gov/pubmed/30232095?dopt=AbstractPlus]). Activation of these receptors produces a wide range of physiological effects throughout the body including the inhibition of secretion of many hormones. Endogenous ligands for these receptors are somatostatin\u201014 ) and somatostatin\u201028 ). http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2007 {Mouse, Rat} has also been suggested to be an endogenous ligand for somatostatin receptors [http://www.ncbi.nlm.nih.gov/pubmed/8622767?dopt=AbstractPlus].Somatostatin (somatotropin release inhibiting factor) is an abundant neuropeptide, which acts on five subtypes of somatostatin receptor , has affinity for SST2, SST4 and SST5 receptors and is a potent inhibitor of GH secretion .. The Succinate receptor has been identified as being activated by physiological levels of the Kreb's cycle intermediate succinate and other dicarboxylic acids such as maleate in 2004. Since its pairing with its endogenous ligand, the receptor has been the focus of intensive research and its role has been evidenced in various (patho)physiological processes such as regulation of renin production, retinal angiogenesis, inflammation or immune response.SUCNR1, one giving a protein of 330 amino acids (AA) and the other one 334\u2010AA. Wittenberger et al. [http://www.ncbi.nlm.nih.gov/pubmed/11273702?dopt=AbstractPlus] noted that the 330\u2010AA protein was more likely to be expressed given the Kozak sequence surrounding the second ATG. Some databases report SUCNR1 as being 334\u2010AA long.In humans, there is the possibility of two open\u2010reading frames (ORFs) for et al. (2012) The succinate receptor as a novel therapeutic target for oxidative and metabolic stress\u2010related conditions. Front Endocrinol (Lausanne)3:22 [https://www.ncbi.nlm.nih.gov/pubmed/22649411?dopt=AbstractPlus]Ariza AC et al. (2016) GPR91: expanding the frontiers of Krebs cycle intermediates. Cell Commun. Signal14:3[https://www.ncbi.nlm.nih.gov/pubmed/26759054?dopt=AbstractPlus]de Castro Fonseca M et al. (2016) Insight into SUCNR1 (GPR91) structure and function. Pharmacol. Ther.159: 56\u201065[https://www.ncbi.nlm.nih.gov/pubmed/26808164?dopt=AbstractPlus]Gilissen J et al. (2018) Multiple faces of succinate beyond metabolism in blood. Haematologica103: 1586\u20101592[https://www.ncbi.nlm.nih.gov/pubmed/29954939?dopt=AbstractPlus]Grimolizzi F NC\u2010IUPHARprovisional nomenclature as recommended by [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) are activated by the endogenous peptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2098 (SP), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2089) , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2090 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2091 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3667 (https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:11517) . The neurokinins (A and B) are mammalian members of the tachykinin family, which includes peptides of mammalian and nonmammalian origin containing the consensus sequence: Phe\u2010x\u2010Gly\u2010LeuMet. Marked species differences in in vitro pharmacology exist for all three receptors, in the context of nonpeptide ligands. Antagonists such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3490 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7623 were approved by FDA and EMA, in combination with other antiemetic agents, for the prevention of nausea and vomiting associated with emetogenic cancer chemotherapy.Tachykinin receptors on the NK1 receptor. There are additional subtypes of tachykinin receptor;an orphan receptor (Swis\u2010sProt http://www.ncbi.nlm.nih.gov/protein/266702/) with structural similarities to the NK3 receptor was found to respond to NKB when expressed in Xenopus oocytes or Chinese hamster ovary cells .The NKet al. (2011) Neurokinin\u20101 receptor: functional significance in the immune system in reference to selected infections and inflammation. Ann. N. Y. Acad. Sci.1217: 83\u201095[https://www.ncbi.nlm.nih.gov/pubmed/21091716?dopt=AbstractPlus]Douglas SD et al. (2005) International Union of Pharmacology. XLVI. G protein\u2010coupled receptor list. Pharmacol Rev57: 279\u2010288[https://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]Foord SM et al. (2008) The neurokinin 1 receptor: a potential new target for anti\u2010platelet therapy? Curr Opin Pharmacol8: 114\u20109[https://www.ncbi.nlm.nih.gov/pubmed/18296119?dopt=AbstractPlus]Jones S et al. (2014) Tachykinins and their receptors: contributions to physiological control and the mechanisms of disease. Physiol. Rev.94: 265\u2010301[https://www.ncbi.nlm.nih.gov/pubmed/24382888?dopt=AbstractPlus]Steinhoff MS et al. (2018) Crystal structure of the human NK_1 tachykinin receptor. Proc. Natl. Acad. Sci. U.S.A.115: 13264\u201013269[https://www.ncbi.nlm.nih.gov/pubmed/30538204?dopt=AbstractPlus]Yin J NC\u2010IUPHARprovisional nomenclature as recommended by [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) are activated by the endogenous tripeptide http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2139 (pGlu\u2010His\u2010ProNH2). http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2139 and TRH analogues fail to distinguish TRH1 and TRH2 receptors [http://www.ncbi.nlm.nih.gov/pubmed/12683933?dopt=AbstractPlus]. [http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3836) is able to label both TRH1 and TRH2 receptors with Kd values of 13 and 9 nM respectively. Synthesis and biology of ring\u2010modified L\u2010Histidine containing TRH analogues has been reported [http://www.ncbi.nlm.nih.gov/pubmed/26854379?dopt=AbstractPlus].Thyrotropin\u2010releasing hormone (TRH) receptors TRH\u2010like peptides. Physiol Res60: 207\u201015[https://www.ncbi.nlm.nih.gov/pubmed/21114375?dopt=AbstractPlus]B\u00edlek R et al. (2005) International Union of Pharmacology. XLVI. G protein\u2010coupled receptor list. Pharmacol Rev57: 279\u2010288[https://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]Foord SM Front Neuroendocrinol31: 134\u201056[https://www.ncbi.nlm.nih.gov/pubmed/20074584?dopt=AbstractPlus]Nillni EA. (2010) Regulation of the hypothalamic thyrotropin releasing hormone (TRH) neuron by neuronal and peripheral inputs. http://www.ncbi.nlm.nih.gov/pubmed/11459929?dopt=AbstractPlus], where 15 mammalian orthologues were identified and divided into two families. The TA1 receptor (nomenclature as agreed by theNC\u2010IUPHAR Subcommittee for the Trace amine receptor [http://www.ncbi.nlm.nih.gov/pubmed/19325074?dopt=AbstractPlus]) has affinity for the endogenous trace amines http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2150, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2144 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2149 in addition to the classical amine http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=940 [http://www.ncbi.nlm.nih.gov/pubmed/11459929?dopt=AbstractPlus]. Emerging evidence suggests that TA1 is a modulator of monoaminergic activity in the brain [http://www.ncbi.nlm.nih.gov/pubmed/19482011?dopt=AbstractPlus] with TA1 and dopamine D2 receptors shown to form constitutive heterodimers when co\u2010expressed [http://www.ncbi.nlm.nih.gov/pubmed/21670104?dopt=AbstractPlus]. In addition to trace amines, receptors can be activated by amphetamine\u2010like psychostimulants, and endogenous thyronamines.Trace amine\u2010associated receptors were discovered from a search for novel 5\u2010HT receptors for detailed discussion. The product of the gene TAAR2 appears to respond to http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2144 > http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2150 and to couple through Gs [http://www.ncbi.nlm.nih.gov/pubmed/11459929?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=115#show_object_168, in some individuals, and http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=115#show_object_169 are pseudogenes in man, although functional in rodents. The signalling characteristics and pharmacology of http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=115#show_object_170 , http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=115#show_object_171http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=115#show_object_171 , http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=115#show_object_172 and http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=115#show_object_173http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=115#show_object_173 are lacking. The thyronamines, endogenous derivatives of thyroid hormone, have affinity for rodent cloned trace amine receptors, including TA1 [http://www.ncbi.nlm.nih.gov/pubmed/15146179?dopt=AbstractPlus]. An antagonist http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5457 has recently been described with a pKi of 9.1 at the mouse TA1 but >5.3 for human TA1 [http://www.ncbi.nlm.nih.gov/pubmed/21237643?dopt=AbstractPlus].In addition to TAet al. (2009) International Union of Pharmacology. LXXII. Recommendations for trace amine receptor nomenclature. Pharmacol. Rev.61: 1\u20108 [https://www.ncbi.nlm.nih.gov/pubmed/19325074?dopt=AbstractPlus]Maguire JJ et al. (2016) Trace Amines and the Trace Amine\u2010Associated Receptor 1: Pharmacology, Neurochemistry, and Clinical Implications. Front Neurosci10: 148 [https://www.ncbi.nlm.nih.gov/pubmed/27092049?dopt=AbstractPlus]Pei Y NC\u2010IUPHAR Subcommittee on the Urotensin receptornomenclature as agreed by the ) is activated by the endogenous dodecapeptide http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2153 , originally isolated from the urophysis, the endocrine organ of the caudal neurosecretory system of teleost fish . Several structural forms of U\u2010II exist in fish and amphibians. The goby orthologue was used to identify U\u2010II as the cognate ligand for the predicted receptor encoded by the rat gene gpr14 . Human http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2153), an 11\u2010amino\u2010acid peptide [http://www.ncbi.nlm.nih.gov/pubmed/9861051?dopt=AbstractPlus], retains the cyclohexapeptide sequence of goby U\u2010II that is thought to beimportant in ligand binding . This sequence is also conserved in the deduced amino\u2010acid sequence of rat http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2155 {Rat} (14 amino\u2010acids) and mouse http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2154 {Mouse} (14 amino\u2010acids), although the N\u2010terminal is more divergent from the human sequence [http://www.ncbi.nlm.nih.gov/pubmed/10486557?dopt=AbstractPlus]. A second endogenous ligand for the UT has been discovered in rat [http://www.ncbi.nlm.nih.gov/pubmed/17628210?dopt=AbstractPlus]. This is the http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2156 , an octapeptide that is derived from a different gene, but shares the C\u2010terminal sequence (CFWKYCV) common to U\u2010II from other species. Identical sequences to rat http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2156) are predicted for the mature mouse and human peptides [http://www.ncbi.nlm.nih.gov/pubmed/18710417?dopt=AbstractPlus]. UT exhibits relatively high sequence identity with somatostatin, opioid and galanin receptors [http://www.ncbi.nlm.nih.gov/pubmed/25535277?dopt=AbstractPlus].The urotensin\u2010II (U\u2010II) receptor elicits both vasoconstrictor and vasodilator responses.In the human vasculature, human et al. (2005) International Union of Pharmacology. XLVI. G protein\u2010coupled receptor list. Pharmacol Rev57: 279\u2010288[https://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]Foord SM et al. (2010) A rat brain atlas of urotensin\u2010II receptor expression and a review of central urotensin\u2010II effects. Naunyn Schmiedebergs Arch. Pharmacol.382:1\u201031[https://www.ncbi.nlm.nih.gov/pubmed/20422157?dopt=AbstractPlus]Hunt BD et al. (2010) Urotensin\u2010II receptor modulators as potential drugs. J. Med. Chem.53: 2695\u2010708[https://www.ncbi.nlm.nih.gov/pubmed/20043680?dopt=AbstractPlus]Maryanoff BE et al. (2010) Role of urotensin II in health and disease. Am.J.Physiol.Regul.Integr.Comp. Physiol.298: R1156\u201072[https://www.ncbi.nlm.nih.gov/pubmed/20421634?dopt=AbstractPlus]Ross B et al. (2015) International Union of Basic and Clinical Pharmacology. XCII. Urotensin II, urotensin II\u2010related peptide, and their receptor: from structure to function. Pharmacol. Rev.67: 214\u201058[https://www.ncbi.nlm.nih.gov/pubmed/25535277?dopt=AbstractPlus]Vaudry H NC\u2010IUPHARnomenclature as recommended by [http://www.ncbi.nlm.nih.gov/pubmed/15914470?dopt=AbstractPlus]) are activated by the endogenous cyclic nonapeptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2168 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2174 . These peptides are derived from precursors which also produce neurophysins . Vasopressin and oxytocin differ at only 2 amino acids (positions 3 and 8). There are metabolites of these neuropeptides that may be biologically active [http://www.ncbi.nlm.nih.gov/pubmed/8258377?dopt=AbstractPlus].Vasopressin (AVP) and oxytocin (OT) receptors exhibit low affinity at human V2 receptors [http://www.ncbi.nlm.nih.gov/pubmed/9773787?dopt=AbstractPlus]. Similarly, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2170 is more V2 selective in the rat than in the human [http://www.ncbi.nlm.nih.gov/pubmed/9264324?dopt=AbstractPlus]. The gene encoding the V2 receptor is polymorphic in man, underlying nephrogenic diabetes insipidus [http://www.ncbi.nlm.nih.gov/pubmed/9756088?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2187 is selective only for the human and bovine V1B receptors [http://www.ncbi.nlm.nih.gov/pubmed/12446593?dopt=AbstractPlus], while http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2191 has high affinity for the rat V1B receptor [http://www.ncbi.nlm.nih.gov/pubmed/17300166?dopt=AbstractPlus]. Knockouts of vasopressin and oxytocin receptors have systemspecific defects which include behavioural deficits .Vasopressin and oxytocin receptors have a characteristic and sometimes overlapping distribution in a number of tissues including brain. There are phylogenetic, ontogenetic and sex\u2010specific differences in the levels and distribution of these receptors, particularly in the brain. The VAm. J. Physiol., Cell Physiol.303: C1115\u201024[https://www.ncbi.nlm.nih.gov/pubmed/22932685?dopt=AbstractPlus]Knepper MA. (2012) Systems biology in physiology: the vasopressin signaling network in kidney. et al. (2012) Vasopressin V1a and V1b receptors: from molecules to physiological systems. Physiol. Rev.92: 1813\u201064[https://www.ncbi.nlm.nih.gov/pubmed/23073632?dopt=AbstractPlus]Koshimizu TA et al. (2012) Oxytocin and vasopressin agonists and antagonists as research tools and potential therapeutics. J. Neuroendocrinol.24: 609\u201028[https://www.ncbi.nlm.nih.gov/pubmed/22375852?dopt=AbstractPlus]Manning M et al. (2011) Oxytocin and vasopressin in the human brain: social neuropeptides for translational medicine. Nat. Rev. Neurosci.12: 524\u201038[https://www.ncbi.nlm.nih.gov/pubmed/21852800?dopt=AbstractPlus]Meyer\u2010Lindenberg A et al. (2012) Balance of brain oxytocin and vasopressin: implications for anxiety, depression, and social behaviors. Trends Neurosci.35: 649\u201059[https://www.ncbi.nlm.nih.gov/pubmed/22974560?dopt=AbstractPlus]Neumann ID NC\u2010IUPHAR Subcommittee on Vasoactive Intestinal Peptide Receptorsnomenclature as agreed by the ) are activated by the endogenous peptides http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1152 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2258 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2257), peptide histidine isoleucineamide , peptide histidine methionineamide (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2274)) and peptide histidine valine (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3706)). VPAC1 and VPAC2 receptors display comparable affinity for the PACAP peptides, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2257) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2258), and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1152 , whereas http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2257 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2258) are >100 fold more potent than http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1152) as agonists of most isoforms of the PAC1 receptor. However, one splice variant of the human PAC1 receptor has been reported to respond to http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2258), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2257) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1152) with comparable affinity [http://www.ncbi.nlm.nih.gov/pubmed/10583729?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2272 [http://www.ncbi.nlm.nih.gov/pubmed/11068102?dopt=AbstractPlus] has been used as a selective VPAC2 receptor antagonist in a number of physiological studies, but has been reported to have significant activity at VPAC1 and PAC1 receptors [http://www.ncbi.nlm.nih.gov/pubmed/16930633?dopt=AbstractPlus]. The selective PAC1 receptor agonist http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2264, was extracted from the salivary glands of sand flies and has no sequence homology to http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1152) or the PACAP peptides [http://www.ncbi.nlm.nih.gov/pubmed/8995389?dopt=AbstractPlus]. Two deletion variants of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2264, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3305 [http://www.ncbi.nlm.nih.gov/pubmed/9928019?dopt=AbstractPlus] and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2265 [http://www.ncbi.nlm.nih.gov/pubmed/10438479?dopt=AbstractPlus] have been reported to be PAC1 receptor antagonists, but these peptides have not been extensively characterised.Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase\u2010activating peptide (PACAP) receptors and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2258); these might result from differences in G protein coupling and second messenger mechanisms [http://www.ncbi.nlm.nih.gov/pubmed/8967982?dopt=AbstractPlus], or from alternative splicing of PAC1 receptor mRNA [http://www.ncbi.nlm.nih.gov/pubmed/8396727?dopt=AbstractPlus].Subtypes of PACet al. (1998) International Union of Pharmacology. XVIII. Nomenclature of receptors for vasoactive intestinal peptide and pituitary adenylate cyclase\u2010activating polypeptide. Pharmacol Rev50: 265\u2010270[https://www.ncbi.nlm.nih.gov/pubmed/9647867?dopt=AbstractPlus]Harmar AJ et al. (2012) Pharmacology and functions of receptors for vasoactive intestinal peptide and pituitary adenylate cyclase\u2010activating polypeptide: IUPHAR review 1. Br. J. Pharmacol.166: 4\u201017[https://www.ncbi.nlm.nih.gov/pubmed/22289055?dopt=AbstractPlus]Harmar AJ et al. (2012) Effects of pituitary adenylate cyclase activating polypeptide in the urinary system, with special emphasis on its protective effects in the kidney. Neuropeptides46: 61\u201070[https://www.ncbi.nlm.nih.gov/pubmed/21621841?dopt=AbstractPlus]Reglodi D et al. (2012) Is PACAP the major neurotransmitter for stress transduction at the adrenomedullary synapse? J. Mol. Neurosci.48: 403\u201012[https://www.ncbi.nlm.nih.gov/pubmed/22610912?dopt=AbstractPlus]Smith CB"} +{"text": "This article has been corrected: The correct author information is given below:Satdarshan P. Monga1475-1490. https://doi.org/10.18632/oncotarget.26668Original article: Oncotarget. 2019; 10:1475\u20131490."} +{"text": "Although the Concise Guide represents approximately 400 pages, the material presented is substantially reduced compared to information and links presented on the website. It provides a permanent, citable, point\u2010in\u2010time record that will survive database updates. The full contents of this section can be found at http://onlinelibrary.wiley.com/doi/10.1111/bph.14752. Enzymes are one of the six major pharmacological targets into which the Guide is divided, with the others being: G protein\u2010coupled receptors, ion channels, nuclear hormone receptors, catalytic receptors and transporters. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading. The landscape format of the Concise Guide is designed to facilitate comparison of related targets from material contemporary to mid\u20102019, and supersedes data presented in the 2017/18, 2015/16 and 2013/14 Concise Guides and previous Guides to Receptors and Channels. It is produced in close conjunction with the International Union of Basic and Clinical Pharmacology Committee on Receptor Nomenclature and Drug Classification (NC\u2010IUPHAR), therefore, providing official IUPHAR classification and nomenclature for human drug targets, where appropriate.The Concise Guide to PHARMACOLOGY 2019/20 is the fourth in this series of biennial publications. The Concise Guide provides concise overviews of the key properties of nearly 1800 human drug targets with an emphasis on selective pharmacology (where available), plus links to the open access knowledgebase source of drug targets and their ligands ( The Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (NC\u2010IUBMB) classifies enzymes into families, using a four number code, on the basis of the reactions they catalyse. There are six main families:EC 1.\u2010.\u2010.\u2010 Oxidoreductases;EC 2.\u2010.\u2010.\u2010 Transferases;EC 3.\u2010.\u2010.\u2010 Hydrolases;EC 4.\u2010.\u2010.\u2010 Lyases;EC 5.\u2010.\u2010.\u2010 Isomerases;EC 6.\u2010.\u2010.\u2010 Ligases.http://www.ncbi.nlm.nih.gov/pubmed/17139284?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/24016212?dopt=AbstractPlus], which is not to say that they are of modest importance.Although there are many more enzymes than receptors in biology, and many drugs that target prokaryotic enzymes are effective medicines, overall the number of enzyme drug targets is relatively small .A number of organophosphorus compounds inhibit acetylcholinesterase and cholinesterase irreversibly, including pesticides such as chlorpyrifos\u2010oxon, and nerve agents such as tabun, soman and sarin. AChE is unusual in its exceptionally high turnover rate which has been calculated at 740 000/min/molecule , which is insensitive to http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5179 [http://www.ncbi.nlm.nih.gov/pubmed/20147294?dopt=AbstractPlus]. Other forms of adenosine deaminase act on ribonucleic acids and may be divided into two families: https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:228 (http://www.uniprot.org/uniprot/Q9BUB4) deaminates transfer RNA; https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:225 ; https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:226 andhttps://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:227 act on double\u2010stranded RNA. Particular polymorphisms of the ADA gene result in loss\u2010of\u2010function and severe combined immunodeficiency syndrome. Adenosine deaminase is able to complex with dipeptidyl peptidase IV to form a cell\u2010surface activity [http://www.ncbi.nlm.nih.gov/pubmed/8101391?dopt=AbstractPlus].An extracellular adenosine deaminase activity, termed ADA2 or adenosine deaminase growth factor Adenosinergic signaling in epilepsy. et al. (2015) Moonlighting adenosine deaminase: a target protein for drug development. Med Res Rev35: 85\u2013125 https://www.ncbi.nlm.nih.gov/pubmed/24933472?dopt=AbstractPlusCort\u00e9s A Nat Rev Mol Cell Biol17: 83\u201396 https://www.ncbi.nlm.nih.gov/pubmed/26648264?dopt=AbstractPlusNishikura K. (2016) A\u2010to\u2010I editing of coding and non\u2010coding RNAs by ADARs. Neuroscience338: 1\u201318 https://www.ncbi.nlm.nih.gov/pubmed/26500181?dopt=AbstractPlusSawynok J. (2016) Adenosine receptor targets for pain. et al. (2015) Role of S\u2010adenosylhomocysteine in cardiovascular disease and its potential epi\u2010genetic mechanism. Int. J. Biochem. Cell Biol. 67: 158\u201366 https://www.ncbi.nlm.nih.gov/pubmed/26117455?dopt=AbstractPlusXiao Y http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5276 as co\u2010substrate and co\u2010factor, respectively. In humans, as well as in other mammals, there are two distinct L\u2010Tryptophan hydroxylase 2 genes. In humans, these genes are located on chromosomes 11 and 12 and encode two different homologous enzymes, TPH1 and TPH2.The amino acid hydroxylases (monooxygenases), EC.1.14.16.\u2010, are iron\u2010containing enzymes which utilise molecular oxygen and et al. (2015) Serotonergic gene variation in substance use pharmacotherapy: a systematic review. Pharmacogenomics16: 1307\u201314 https://www.ncbi.nlm.nih.gov/pubmed/26265436?dopt=AbstractPlusBauer IE et al. (2011) Tyrosine hydroxylase and regulation of dopamine synthesis. Arch Biochem Biophys508: 1\u201312 https://www.ncbi.nlm.nih.gov/pubmed/21176768?dopt=AbstractPlusDaubner SC et al. (2013) Phenylalanine hydroxylase: function, structure, and regulation. IUBMB Life65: 341\u20139 https://www.ncbi.nlm.nih.gov/pubmed/23457044?dopt=AbstractPlusFlydal MI et al. (2013) Mechanisms of tryptophan and tyrosine hydroxylase. IUBMB Life65: 350\u20137 https://www.ncbi.nlm.nih.gov/pubmed/23441081?dopt=AbstractPlusRoberts KM et al. (2014) Complex molecular regulation of tyrosine hydroxylase. J Neural Transm121: 1451\u201381 https://www.ncbi.nlm.nih.gov/pubmed/24866693?dopt=AbstractPlusTekin I et al. (2017) Tyrosine and tryptophan hydroxylases as therapeutic targets in human disease. Expert Opin Ther Targets21: 167\u2013180 https://www.ncbi.nlm.nih.gov/pubmed/27973928?dopt=AbstractPlusWalen K http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=721 is a basic amino acid with a guanidino sidechain. As an amino acid, metabolism of L\u2010arginine to form http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=725, catalysed by arginase, forms the last step of the urea production cycle. L\u2010Ornithine may be utilised as a precursor of polyamines (see http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=240) or recycled via http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5324 to L\u2010arginine. L\u2010Arginine http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=240#Decarboxylases to form http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4127, although the prominence of this pathway in human tissues is uncertain. L\u2010Arginine may be used as a precursor for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5325 formation in the http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4496 synthesis pathway under the influence of arginine:glycine amidinotransferase with L\u2010ornithine as a byproduct. Nitric oxide synthase uses L\u2010arginine to generate nitric oxide, with http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=722 also as a byproduct. L\u2010Arginine in proteins may be subject to post\u2010translational modification through methylation, catalysed by protein arginine methyltransferases. Subsequent proteolysis can liberate asymmetric http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5229 (ADMA), which is an endogenous inhibitor of nitric oxide synthase activities. ADMA is hydrolysed by dimethylarginine dimethylhydrolase activities to generate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=722 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5177.N\u2010methyltransferases encompass histone arginine N\u2010methyltransferases and myelin basic protein N\u2010methyltransferases . They are dimeric or tetrameric enzymes which use http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4786 as a methyl donor, generating http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5265 as a by\u2010product. They generate both mono\u2010methylated and di\u2010methylated products; these may be symmetric (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5271) or asymmetric (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5229) versions, where both guanidine nitrogens are monomethylated or one of the two is dimethylated, respectively.Protein arginine http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=254.Information on members of this family may be found in the http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=3.5.3.1) are manganese\u2010containing isoforms, which appear to show differential distribution, where the ARG1 isoform predominates in the liver and erythrocytes, while ARG2 is associated more with the kidney.Arginase are cytoplasmic enzymes which hydrolyse http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5229 to form http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5177 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=722.Dimethylarginine dimethylaminohydrolases are a family of oxidoreductases that synthesize nitric oxide (NO.) via the NADPH and oxygen\u2010dependent consumption of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=721 with the resultant by\u2010product, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=722. There are 3 NOS isoforms and they are related by their capacity to produce NO, highly conserved organization of functional domains and significant homology at the amino acid level. NOS isoforms are functionally distinguished by the cell type where they are expressed, intracellular targeting and transcriptional and post\u2010translation mechanisms regulating enzyme activity. The nomenclature suggested by NC\u2010IUPHAR of NOS I, II and III [http://www.ncbi.nlm.nih.gov/pubmed/9228663?dopt=AbstractPlus] has not gained wide acceptance, and the 3 isoforms are more commonly referred to as neuronal NOS (nNOS), inducible NOS (iNOS) and endothelial NOS (eNOS) which reflect the location of expression (nNOS and eNOS) and inducible expression (iNOS). All are dimeric enzymes that shuttle electrons from NADPH, which binds to a C\u2010terminal reductase domain, through the flavins FAD and FMN to the oxygenase domain of the other monomer to enable the BH4\u2010dependent reduction of heme bound oxygen for insertion into the substrate, L\u2010arginine. Electron flow from reductase to oxygenase domain is controlled by calmodulin binding to canonical calmodulin binding motif located between these domains. eNOS and nNOS isoforms are activated at concentrations of calcium greater than 100 nM, while iNOS shows higher affinity for Ca2+/http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2351 with great avidity and is essentially calcium\u2010independent and constitutively active. Efficient stimulus\u2010dependent coupling of nNOS and eNOS is achieved via subcellular targeting through respective N\u2010terminal PDZ and fatty acid acylation domains whereas iNOS is largely cytosolic and function is independent of intracellular location. nNOS is primarily expressed in the brain and neuronal tissue, iNOS in immune cells such as macrophages and eNOS in the endothelial layer of the vasculature although exceptions in other cells have been documented. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5213 and related modified arginine analogues are inhibitors of all three isoforms, with IC50 values in the micromolar range.Nitric oxide synthases Nitric Oxide Signaling in T Cell\u2010Mediated Immunity et al. (2015) Strategies to increase nitric oxide signalling in cardiovascular disease. Nat Rev Drug Discov14: 623\u201341 https://www.ncbi.nlm.nih.gov/pubmed/26265312?dopt=AbstractPlusLundberg JO et al. (2016) Endothelial nitric oxide synthase: From biochemistry and gene structure to clinical implications of NOS3 polymorphisms. Gene575: 584\u201399 https://www.ncbi.nlm.nih.gov/pubmed/26428312?dopt=AbstractPlusOliveira\u2010Paula GH BrJPharmacol176: 177\u2013188 https://www.ncbi.nlm.nih.gov/pubmed/30402946Stuehr DJ and Haque MM (2019) Nitric oxide synthase enzymology in the 20 years after the Nobel Prize. Br J Pharmacol176: 147\u2013154 https://www.ncbi.nlm.nih.gov/pubmed/30357812Wallace JL (2019) Nitric oxide in the gastrointestinal tract: opportunities for drug development. et al. (2016) Modulating DDAH/NOS Pathway to Discover Vasoprotective Insulin Sensitizers. J Diabetes Res2016: 1982096 https://www.ncbi.nlm.nih.gov/pubmed/26770984?dopt=AbstractPlusLai L et al. (1997) International Union of Pharmacology Nomenclature in Nitric Oxide Re\u2010search. Pharmacol. Rev. 49: 137\u201342 https://www.ncbi.nlm.nih.gov/pubmed/9228663?dopt=AbstractPlusMoncada S et al. (2015) The crucial role of l\u2010arginine in macrophage activation: What you need to know about it. Life Sci. 137: 44\u20138 https://www.ncbi.nlm.nih.gov/pubmed/26188591?dopt=AbstractPlusPekarova M et al. (2017) Arginase Inhibitors: A Rational Approach Over One Century. Med Res Rev37: 475\u2013513 https://www.ncbi.nlm.nih.gov/pubmed/27862081?dopt=AbstractPlusPudlo M et al. (2016) Benefits of L\u2010Arginine on Cardiovascular System. Mini Rev Med Chem16: 94\u2013103 https://www.ncbi.nlm.nih.gov/pubmed/26471966?dopt=AbstractPlusSudar\u2010Milovanovic E Carbonic anhydrases facilitate the interconversion of water and carbon dioxide with bicarbonate ions and protons (EC 4.2.1.1), with over a dozen gene products identified in man. The enzymes function in acid\u2010base balance and the movement of carbon dioxide and water. They are targetted for therapeutic gain by particular antiglaucoma agents and diuretics.Bioorg Med Chem21: 1570\u201370 https://www.ncbi.nlm.nih.gov/pubmed/22607884Imtaiyaz Hassan M, Shajee B, Waheed A, Ahmad F and Sly WS. (2013) Structure, function and applications of carbonic anhydrase isozymes. Expert Opin Drug Discov12: 61\u201388 https://www.ncbi.nlm.nih.gov/pubmed/27783541Supuran CT (2017) Advances in structure\u2010based drug discovery of carbonic anhydrase inhibitors. Future Med Chem10: 561\u2013573 https://www.ncbi.nlm.nih.gov/pubmed/29478330Supuran CT (2018) Carbonic anhydrase activators. 3\u2010 or CO2 into target molecules. Two groups of carboxylase activities, some of which are bidirectional, can be defined on the basis of the cofactor requirement, making use of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4787 (EC 6.4.1.\u2010) or http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5286 (EC 4.1.1.\u2010).The carboxylases allow the production of new carbon\u2010carbon bonds by introducing HCOhttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2478 are able to activate ACC1/ACC2 activity allosterically. PCC is able to function in forward and reverse modes as a ligase (carboxylase) or lyase (decarboxylase) activity, respectively. Loss\u2010of\u2010function mutations in GGCX are associated with clotting disorders.Dicarboxylic acids including 2 and the indicated products from acidic substrates, requiring http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5249 or http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4809 as a co\u2010factor.The decarboxylases generate COet al. (2010) Structural biology of S\u2010adenosylmethionine decarboxylase. Amino Acids38: 451\u201360 . Complexes involving SPT3 appeared more broad in substrate selectivity, with incorporation of myristoylCoA prominent for SPT1/SPT3/ssSPTa complexes, while SP1/SPT3/ssSPTb complexes had similar activity with C16, C18 and C20 acylCoAs [http://www.ncbi.nlm.nih.gov/pubmed/19416851?dopt=AbstractPlus].The functional enzyme is a heterodimer of SPT1 (LCB1) with either SPT2 (LCB2) or SPT3 (LCB2B); the small subunits of SPT (ssSPTa or ssSPTb) bind to the heterodimer to enhance enzymatic activity. The complexes of SPT1/SPT2/ssSPTa and SPT1/SPT2/ssSPTb were most active with palmitoylCoA as substrate, with the latter complex also showing some activity with stearoylCoA .DEGS1 activity is inhibited by a number of natural products, including Following translocation from the ER to the Golgi under the influence of the ceramide transfer protein, sphingomyelin synthases allow the formation of sphingomyelin by the transfer of phosphocholine from the phospholipid phosphatidylcholine. Sphingomyelin synthase\u2010related protein 1 is structurally related but lacks sphingomyelin synthase activity.Also known as sphingomyelinase.http://www.ncbi.nlm.nih.gov/pubmed/8808629?dopt=AbstractPlus] and polycomb protein EED [http://www.ncbi.nlm.nih.gov/pubmed/20080539?dopt=AbstractPlus] allow coupling between TNF receptors and neutral sphingomyelinase phosphodiesterases.Protein FAN . In normal cells, tissues and organs, precise co\u2010ordination of these proteins ensures expression of only those genes required to specify phenotype or which are required at specific times, for specific functions. Chromatin modifications allow DNA modifications not coded by the DNA sequence to be passed on through the genome and underlies heritable phenomena such as X chromosome inactivation, aging, heterochromatin formation, reprogramming, and gene silencing (epigenetic control).Chromatin modifying enzymes, and other chromatin\u2010modifying proteins, fall into three broad categories: http://www.ncbi.nlm.nih.gov/pubmed/17320507?dopt=AbstractPlus].To date at least eight distinct types of modifications are found on histones. These include small covalent modifications such as acetylation, methylation, and phosphorylation, the attachment of larger modifiers such as ubiquitination or sumoylation, and ADP ribosylation, proline isomerization and deimination. Chromatin modifications and the functions they regulate in cells are reviewed by Kouzarides (2007) , where awide variety of cellular and protein abberations are known to perturb chromatin structure, gene transcription and ultimately cellular pathways . Due to the reversible nature of epigenetic modifications, chromatin regulators are very tractable targets for drug discovery and the development of novel therapeutics. Indeed, small molecule inhibitors of writers and erasers are already being used in the clinic. The search for the next generation of compounds with improved specificity against chromatin\u2010associated proteins is an area of intense basic and clinical research [http://www.ncbi.nlm.nih.gov/pubmed/25974248?dopt=AbstractPlus]. Current progress in this field is reviewed by Sim\u00f3\u2010Riudalbas and Esteller (2015) [http://www.ncbi.nlm.nih.gov/pubmed/25039449?dopt=AbstractPlus].Dysregulated epigenetic control can be associated with human diseases such as cancer .Classes I, II and IV use Znhttp://www.ncbi.nlm.nih.gov/pubmed/19608861?dopt=AbstractPlus] such as microtubules [http://www.ncbi.nlm.nih.gov/pubmed/12024216?dopt=AbstractPlus], the hsp90 chaperone [http://www.ncbi.nlm.nih.gov/pubmed/15916966?dopt=AbstractPlus] and the tumour suppressor p53 [http://www.ncbi.nlm.nih.gov/pubmed/11099047?dopt=AbstractPlus].HDACs have more general protein deacetylase activity, being able to deacetylate lysine residues in non\u2010histone proteins , making HDACs attractive molecular targets in the search for novel mechanisms to treat cancer [http://www.ncbi.nlm.nih.gov/pubmed/24382387?dopt=AbstractPlus]. Several small molecule HDAC inhibitors are already approved for clinical use: http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7006, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7496, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6852, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7489, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7496, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7009 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=8305. HDACs and HDAC inhibitors currently in development as potential anti\u2010cancer therapeutics are reviewed by Sim\u00f3\u2010Riudalbas and Esteller (2015) [http://www.ncbi.nlm.nih.gov/pubmed/25039449?dopt=AbstractPlus].Dysregulated HDACactivity has been identified in cancer cells and tumour tissues . Four families of membranous adenylyl cyclase are distinguishable: http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2351 \u2010stimulated , Ca2+\u2010 and G\u03b2\u03b3\u2010inhibitable , G\u03b2\u03b3\u2010stimulated and Ca2+\u2010insensitive , and forskolin\u2010insensitive (AC9) forms. A soluble adenylyl cyclase (AC10) lacks membrane spanning regions and is insensitive to G proteins.It functions as a cytoplasmic bicarbonate (pH\u2010insensitive) sensor [http://www.ncbi.nlm.nih.gov/pubmed/10915626?dopt=AbstractPlus].Adenylyl cyclase, http://www.ncbi.nlm.nih.gov/pubmed/24006339?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/24008337?dopt=AbstractPlus]. AC3 shows only modest in vitro activation by Ca2+/CaM.Many of the activators and inhibitors listed are only somewhat selective or have not been tested against all AC isoforms . Once activated, Epacs induce an enhanced activity of the monomeric G proteins, Rap1 and Rap2 by facilitating binding of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1742 in place of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2410, leading to activation of http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=244 [http://www.ncbi.nlm.nih.gov/pubmed/11715024?dopt=AbstractPlus].Epacs are members of a family of guanine nucleotide exchange factors The role of Epac in the heart. Cell. Mol. Life Sci. 74: 591\u2010606 https://www.ncbi.nlm.nih.gov/pubmed/27549789?dopt=AbstractPlusFujita T Physiol Rev98: 919\u20101053 https://www.ncbi.nlm.nih.gov/pubmed/29537337Robichaux WG and Cheng X. (2018) Intracellular cAMP Sensor EPAC: Physiology, Pathophysiology, and Therapeutics Development. et al. (2017) Exchange proteins directly activated by cAMP (EPACs): Emerging therapeutic targets. Bioorg. Med. Chem. Lett. 27: 1633\u20101639 https://www.ncbi.nlm.nih.gov/pubmed/28283242?dopt=AbstractPlusWang P http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=3.1.4.17, catalyse the hydrolysis of a 3\u2019,5\u2019\u2010cyclic nucleotide . http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=388 is a nonselective inhibitor with an IC50 value in the millimolar range for all isoforms except PDE 8A, 8B and 9A. A 2\u2019,3\u2019\u2010cyclic nucleotide 3\u2019\u2010phosphodiesterase (http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=3.1.4.37 CNPase) activity is associated with myelin formation in the development of the CNS.3\u2019,5\u2019\u2010Cyclic nucleotide phosphodiesterases , PDE1A, 1B and 1C appear to act as soluble homodimers, while PDE2A is a membrane\u2010bound homodimer. PDE3A and PDE3B are membrane\u2010bound.http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2352 specific. The potency of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5292 at other members of the PDE4 family has not been reported. PDE4B\u2010D long forms are inhibited by extracellular signal\u2010regulated kinase (ERK)\u2010mediated phosphorylation . PDE4A\u2010D splice variants can be membrane\u2010bound or cytosolic [http://www.ncbi.nlm.nih.gov/pubmed/12444918?dopt=AbstractPlus]. PDE4 isoforms may be labelled with http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5313.PDE4 isoforms are essentially http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2347 specific and is activated by the a\u2010subunit of transducin (Gat) and inhibited by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4743, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2919 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4807 with potencies lower than those observed for PDE5A. Defects in PDE6B are a cause of retinitis pigmentosa and congenital stationary night blindness.PDE6 is a membrane\u2010bound tetramer composed of two catalytic chains (PDE6A or PDE6C and PDE6B), an inhibitory chain (PDE6G or PDE6H) and the PDE6D chain. The enzyme is essentially Cell. Signal. 28: 713\u20108 https://www.ncbi.nlm.nih.gov/pubmed/26498857?dopt=AbstractPlusKlussmann E. (2016) Protein\u2010protein interactions of PDE4 family members \u2010 Functions, interactions and therapeutic value. et al. (2018) Targeting phosphodiesterase 5 as a therapeutic option against myocardial ischaemia/reperfusion injury and for treating heart failure. Br. J. Pharmacol. 175: 223\u2010231 https://www.ncbi.nlm.nih.gov/pubmed/28213937?dopt=AbstractPlusKorkmaz\u2010Ic\u00f6z S et al. (2018) Phosphodiesterase\u20104 Inhibitors for the Treatment of Inflammatory Diseases. Front Pharmacol9: 1048 https://www.ncbi.nlm.nih.gov/pubmed/30386231Li H Life Sci230: 150\u2010161 https://www.ncbi.nlm.nih.gov/pubmed/31125564Mehta A and Patel BM. (2019) Therapeutic opportunities in colon cancer: Focus on phosphodiesterase inhibitors. et al. (2019) Experimental and investigational phosphodiesterase inhibitors in development for asthma. Expert Opin Investig Drugs28: 261\u2010266 https://www.ncbi.nlm.nih.gov/pubmed/30678501Ntontsi P J Cereb Blood Flow Metab38: 189\u2010203 https://www.ncbi.nlm.nih.gov/pubmed/29256324Pauls MM. (2018) The effect of phosphodiesterase\u20105 inhibitors on cerebral blood flow in humans: A systematic review. et al. (2018) Inhibitors of phosphodiesterase as cancer therapeutics. Eur J Med Chem150: 742\u2010756 https://www.ncbi.nlm.nih.gov/pubmed/29574203Peng T et al. (2018) Fragment\u2010Based Drug Discovery of Phosphodiesterase Inhibitors. J Med Chem61: 1415\u20131424 https://www.ncbi.nlm.nih.gov/pubmed/28800229Svensson F et al. (2018) Role of cAMP and phosphodiesterase signaling in liver health and disease. Cell Signal49: 105\u2010115 https://www.ncbi.nlm.nih.gov/pubmed/29902522Wahlang B et al. (2018) Phosphodiesterase 10 Inhibitors \u2010 Novel Perspectives for Psychiatric and Neurodegenerative Drug Discovery. Curr Med Chem25: 3455\u20103481 https://www.ncbi.nlm.nih.gov/pubmed/29521210Zagorska A The cytochrome P450 enzyme family (CYP450), E.C. 1.14.\u2010.\u2010, were originally defined by their strong absorbance at 450 nm due to the reduced carbon monoxide\u2010complexed haem component of the cytochromes. They are an extensive family of haemcontaining monooxygenases with a huge range of both endogenous and exogenous substrates. These include sterols, fat\u2010soluble vitamins, pesticides and carcinogens as well as drugs. The substrates of some orphan CYP are not known. Listed below are the human enzymes; their relationship with rodent CYP450 enzyme activities is obscure in that the species orthologuemay not catalyse the metabolism of the same substrates. Although the majority of CYP450 enzyme activities are concentrated in the liver, the extrahepatic enzyme activities also contribute to patho/physiological processes. Genetic variation of CYP450 isoforms is widespread and likely underlies a significant proportion of the individual variation to drug administration.et al. (2016) Role of Cytochrome P450 2C8 in Drug Metabolism and Interactions. Pharmacol Rev68: 168\u2010241 https://www.ncbi.nlm.nih.gov/pubmed/26721703?dopt=AbstractPlusBackman JT et al. (2017) Cytochrome P450 eicosanoids in cerebrovascular function and disease. Pharmacol Ther179: 31\u201046 https://www.ncbi.nlm.nih.gov/pubmed/28527918?dopt=AbstractPlusDavis CM et al. (2016) Recent Progress in the Discovery of Next Generation Inhibitors of Aromatase from the Structure\u2010Function Perspective. J Med Chem. 59: 5131\u201048 https://www.ncbi.nlm.nih.gov/pubmed/26689671?dopt=AbstractPlusGhosh D et al. (2015) Cytochrome P450 1 family and cancers. J Steroid Biochem Mol Biol. 147: 24\u201030 https://www.ncbi.nlm.nih.gov/pubmed/25448748?dopt=AbstractPlusGo RE et al. (2016) Recent Structural Insights into Cytochrome P450 Function. Trends Pharmacol Sci37: 625\u2010640 https://www.ncbi.nlm.nih.gov/pubmed/27267697?dopt=AbstractPlusGuengerich FP Pharmacol Ther192: 1\u201019 https://www.ncbi.nlm.nih.gov/pubmed/29964123?dopt=AbstractPlusImig JD. (2018) Prospective for cytochrome P450 epoxygenase cardiovascular and renal therapeutics. et al. (2017) Pharmacogenomics of the cytochrome P450 2C family: impacts of amino acid variations on drug metabolism. Drug Discov Today22: 366\u2010376 https://www.ncbi.nlm.nih.gov/pubmed/27693711?dopt=AbstractPlusIsvoran A et al. (2017) Cytochrome P450\u2010derived eicosanoids and heart function. Pharmacol Ther179: 47\u201383 https://www.ncbi.nlm.nih.gov/pubmed/28551025?dopt=AbstractPlusJamieson KL et al. (2018) Spectroscopic studies of the cytochrome P450 reaction mechanisms. Biochim Biophys Acta1866: 178\u2010204 https://www.ncbi.nlm.nih.gov/pubmed/28668640?dopt=AbstractPlusMak PJ et al. (2016) Cholesterol 24\u2010hydroxylase: Brain cholesterol metabolism and beyond. Biochim Biophys Acta1861: 1911\u20101920 https://www.ncbi.nlm.nih.gov/pubmed/27663182?dopt=AbstractPlusMoutinho M et al. (2018) Keeping the spotlight on cytochrome P450. Biochim Biophys Acta1866: 80\u201087 https://www.ncbi.nlm.nih.gov/pubmed/28599858?dopt=AbstractPlusShalan H DNA topoisomerases regulate the supercoiling of nuclear DNA to influence the capacity for replication or transcription. The enzymatic function of this series of enzymes involves cutting the DNA to allow unwinding, followed by re\u2010attachment to reseal the backbone. Members of the family are targetted in anti\u2010cancer chemotherapy.et al. (2017) Topoisomerases: Resistance versus Sensitivity, How Far We Can Go? Med Res Rev37: 404\u2010438 https://www.ncbi.nlm.nih.gov/pubmed/27687257?dopt=AbstractPlusBansal S et al. (2017) Type I DNA Topoisomerases. J. Med. Chem. 60: 2169\u20102192 https://www.ncbi.nlm.nih.gov/pubmed/28072526?dopt=AbstractPlusCapranico G et al. (2017) DNA topoisomerase I and DNA gyrase as targets for TB therapy. Drug Discov. Today22: 510\u2010518 https://www.ncbi.nlm.nih.gov/pubmed/27856347?dopt=AbstractPlusNagaraja V et al. (2016) Roles of eukaryotic topoisomerases in transcription, replication and genomic stability. Nat. Rev. Mol. Cell Biol. 17: 703\u2010721 https://www.ncbi.nlm.nih.gov/pubmed/27649880?dopt=AbstractPlusPommier Y et al. (2016) The dynamic interplay between DNA topoisomerases and DNA topology. Biophys Rev8: 101\u2010111 https://www.ncbi.nlm.nih.gov/pubmed/28510219?dopt=AbstractPlusSeol Y http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=729 (2\u2010AG), and N\u2010acylethanolamines, such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2364 . The glycerol esters and ethanolamides are synthesised and hydrolysed by parallel, independent pathways. Mechanisms for release and re\u2010uptake of endocannabinoids are unclear, although potent and selective inhibitors of facilitated diffusion of endocannabinoids across cell membranes have been developed [http://www.ncbi.nlm.nih.gov/pubmed/29531087?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/ObjectDisplayForward?objectId=2535 (https://www.uniprot.org/uniprot/Q01469) has been suggested to act as a canonical intracellular endocannabinoid transporter in vivo [http://www.ncbi.nlm.nih.gov/pubmed/28584105?dopt=AbstractPlus]. For the generation of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=729, the key enzyme involved is diacylglycerol lipase (DAGL), whilst several routes for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2364 synthesis have been described, the best characterized of which involves N\u2010acylphosphatidylethanolamine\u2010phospholipase D . A transacylation enzyme which forms N\u2010acylphosphatidylethanolamines has been identified as a cytosolic enzyme, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:24791 (http://www.uniprot.org/uniprot/Q3MJ16) [http://www.ncbi.nlm.nih.gov/pubmed/27399000?dopt=AbstractPlus]. In vitro experiments indicate that the endocannabinoids are also substrates for oxidative metabolism via cyclooxygenase, lipoxygenase and cytochrome P450 enzyme activities .The principle endocannabinoids are 2\u2010acylglycerol esters, such as N\u2010acylethanolamine biosynthesis other than through NAPE\u2010PLD activity have been identified [http://www.ncbi.nlm.nih.gov/pubmed/23394527?dopt=AbstractPlus].Routes for http://www.ncbi.nlm.nih.gov/pubmed/22969151?dopt=AbstractPlus], but may also regulate lysophosphatidylserine levels [http://www.ncbi.nlm.nih.gov/pubmed/25580854?dopt=AbstractPlus]. Loss\u2010of\u2010function mutations in ABHD12 are associated with a disorder known as PHARC [http://www.ncbi.nlm.nih.gov/pubmed/20797687?dopt=AbstractPlus].ABHD12 is a 398\u2010aa protein, with serine hydrolase activity. It has a molecular weight of 45 kDa. A single TM is predicted at 75\u201095, with an extracellular catalytic domain. ABHD12 is a monoacylglycerol hydrolase [http://www.ncbi.nlm.nih.gov/pubmed/17015445?dopt=AbstractPlus] and a limited range of inhibitors have been assessed at this enzyme activity. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=729 has been reported to be hydrolysed by multiple enzyme activities from neural preparations [http://www.ncbi.nlm.nih.gov/pubmed/29751000?dopt=AbstractPlus], including https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:21398 (http://www.uniprot.org/uniprot/Q9BV23) [http://www.ncbi.nlm.nih.gov/pubmed/26989199?dopt=AbstractPlus] and carboxylesterase 1 . https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:21398 (http://www.uniprot.org/uniprot/Q9BV23) has also been described as a triacylglycerol lipase and ester hydrolase [http://www.ncbi.nlm.nih.gov/pubmed/27247428?dopt=AbstractPlus], while https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:15868 (http://www.uniprot.org/uniprot/Q8N2K0) is also able to hydrolyse lysophosphatidylserine [http://www.ncbi.nlm.nih.gov/pubmed/2397193?dopt=AbstractPlus]. https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:15868 (http://www.uniprot.org/uniprot/Q8N2K0) has been described to be inhibited selectively by pentacyclic triterpenoids, such as oleanolic acid [http://www.ncbi.nlm.nih.gov/pubmed/24879289?dopt=AbstractPlus].Many of the compounds described as inhibitors are irreversible and so potency estimates will vary with incubation time. FAAH2 is not found in rodents .http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2391 as substrate, these products are hydroperoxyeicosatetraenoic acids (HPETEs). In humans there are five lipoxygenases, the 5S\u2010(arachidonate : oxygen 5\u2010oxidoreductase), 12R\u2010, 12S\u2010(arachidonate : oxygen 12\u2010oxidoreductase), and two distinct 15S\u2010(arachidonate : oxygen 15\u2010oxidoreductase) LOXs that oxygenate arachidonic acid in different positions along the carbon chain and form the corresponding 5S\u2010, 12S\u2010, 12R\u2010, or 15S\u2010hydroperoxides, respectively.The lipoxygenases (LOXs) are a structurally related family of non\u2010heme iron dioxygenases that function in the production, and in some cases metabolism, of fatty acid hydroperoxides. For http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=1.13.11.40, arachidonate:oxygen 8\u2010oxidoreductase) may be the mouse orthologue of 15\u2010LOX\u20102 [http://www.ncbi.nlm.nih.gov/pubmed/12432921?dopt=AbstractPlus]. Some general LOX inhibitors are http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4265 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5180. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5297 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5155 are used as 5\u2010lipoxygenase inhibitors, while http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5144 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5162 are 12\u2010lipoxygenase inhibitors. The specificity of these inhibitors has not been rigorously assessed with all LOX forms: http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5144, along with other flavonoids, such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5182 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5215, also inhibits 15\u2010LOX\u20101 [http://www.ncbi.nlm.nih.gov/pubmed/12628491?dopt=AbstractPlus].An 8\u2010LOX(4 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5214), produced by 5\u2010LOX activity, and lipoxins may be subject to further oxidative metabolism; \u03c9\u2010hydroxylation is mediated by http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=242#show_object_1344 and http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=242#show_object_1345, while \u00df\u2010oxidation in mitochondria and peroxisomes proceeds in a manner dependent on coenzyme A conjugation. Conjugation of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5214 at the 6 position with reduced glutathione to generate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3354 occurs under the influence of leukotriene C4 synthase, with the subsequent formation of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3353 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3352, all three of which are agonists at http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=35. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3353 formation is catalysed by \u03b3\u2010glutamyltransferase, and subsequently dipeptidase 2 removes the terminal http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=727 from http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3353 to generate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3352. Leukotriene A4 hydrolase converts the 5,6\u2010epoxide LTA4 to the 5\u2010hydroxylated LTB4, an agonist for http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=35. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5214 is also acted upon by 12S\u2010LOX to produce the trihydroxyeicosatetraenoic acids lipoxins http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1034 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5216. Treatment with a LTA4 hydrolase inhibitor in a murine model of allergic airway inflammation increased LXA4 levels, in addition to reducing http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2487, in lung lavage fluid [http://www.ncbi.nlm.nih.gov/pubmed/20110560?dopt=AbstractPlus]. LTA4 hydrolase is also involved in biosynthesis of http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=134. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4139 has been reported to increase endogenous formation of 18S\u2010hydroxyeicosapentaenoate (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5105) compared with http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3400, a resolvin precursor. Both enantiomers may be metabolised by human recombinant 5\u2010LOX; recombinant LTA4 hydrolase converted chiral 5S(6)\u2010epoxide\u2010containing intermediates to http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3333 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5106 [http://www.ncbi.nlm.nih.gov/pubmed/21206090?dopt=AbstractPlus].Leukotriene Ahttp://www.ensembl.org/Homo_sapiens/Gene/Family/Genes?family=ENSFM00250000001675), which also includes aminopeptidase B and aminopeptidase B\u2010like 1 . Dipeptidase 1 and 2 are members of a family of membrane dipeptidases, which also includes for which http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3353 appears not to be a substrate.LTA4H is a member of a family of arginyl aminopeptidases The double\u2010edged role of 12/15\u2010lipoxygenase during inflammation and immunity. Biochim. Biophys. Acta1862: 371\u2013381 https://www.ncbi.nlm.nih.gov/pubmed/27480217?dopt=AbstractPlusAckermann JA et al. (2017) The Cardiovascular Pharmacology of Nonsteroidal Anti\u2010Inflammatory Drugs. Trends Pharmacol. Sci. 38: 733\u2013748 https://www.ncbi.nlm.nih.gov/pubmed/28651847?dopt=AbstractPlusGrosser T J Clin Invest128: 2680\u20132690 https://www.ncbi.nlm.nih.gov/pubmed/30108195Haeggstrom JZ. (2018) Leukotriene biosynthetic enzymes as therapeutic targets. et al. (2019) Beyond leukotriene formation\u2010The noncanonical functions of 5\u2010lipoxygenase. Prostaglandins Other Lipid Mediat142: 24\u201332 https://www.ncbi.nlm.nih.gov/pubmed/30930090Hafner AK Br J Pharmacol176: 1038\u20131050 https://www.ncbi.nlm.nih.gov/pubmed/29468666Mitchell JA and Kirkby NS. (2019) Eicosanoids, prostacyclin and cyclooxygenase in the cardiovascular system. et al. (2015) Perspective of microsomal prostaglandin E2 synthase\u20101 as drug target in inflammation\u2010related disorders. Biochem. Pharmacol. 98: 1\u201315 https://www.ncbi.nlm.nih.gov/pubmed/26123522?dopt=AbstractPlusKoeberle A et al. (2015) Mammalian lipoxygenases and their biological relevance. Biochim. Biophys. Acta1851: 308\u201330 https://www.ncbi.nlm.nih.gov/pubmed/25316652?dopt=AbstractPlusKuhn H et al. (2015) Cyclooxygenase inhibitors: From pharmacology to clinical read\u2010outs. Biochim. Biophys. Acta1851: 422\u201332 https://www.ncbi.nlm.nih.gov/pubmed/25263946?dopt=AbstractPlusPatrignani P et al. (2015) 5\u2010Lipoxygenase, a key enzyme for leukotriene biosynthesis in health and disease. Biochim. Biophys. Acta1851: 331\u20139 https://www.ncbi.nlm.nih.gov/pubmed/25152163?dopt=AbstractPlusR\u00e5dmark O et al. (2017) Role of prostacyclin synthase in carcinogenesis. Prostaglandins Other Lipid Mediat. 133: 49\u201352 https://www.ncbi.nlm.nih.gov/pubmed/28506876?dopt=AbstractPlusSasaki Y et al. (2017) Prostaglandin synthases: Molecular characterization and involvement in prostaglandin biosynthesis. Prog. Lipid Res. 66: 50\u201368 https://www.ncbi.nlm.nih.gov/pubmed/28392405?dopt=AbstractPlusSeo MJ et al. (2016) COX\u20101 Inhibitors: Beyond Structure Toward Therapy. Med Res Rev36: 641\u201371 https://www.ncbi.nlm.nih.gov/pubmed/27111555?dopt=AbstractPlusVitale P http://www.ncbi.nlm.nih.gov/pubmed/8126575?dopt=AbstractPlus] where GABA is principally accumulated in vesicles through the action of the vesicular inhibitory amino acid transporter http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=219. The role of \u03b3\u2010aminobutyraldehyde dehydrogenase (ALDH9A1) in neurotransmitter GABA synthesis is less clear. Following release from neurons, GABA may interact with either GABAA or GABAB receptors and may be accumulated in neurones and glia through the action of members of the http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=144. Successive metabolism through GABA transaminase and succinate semialdehyde dehydrogenase generates succinic acid, which may be further metabolized in the mitochondria in the tricarboxylic acid cycle.The inhibitory neurotransmitter \u03b3\u2010aminobutyrate is generated in neurones by glutamic acid decarboxylase. GAD1 and GAD2 are differentially expressed during development, whereGAD2 is thought to subserve a trophic role in early life and is distributed throughout the cytoplasm. GAD1 is expressed in later life and is more associated with nerve terminals , although this mechanism of action has been questioned [http://www.ncbi.nlm.nih.gov/pubmed/15302681?dopt=AbstractPlus]. The aminosteroid http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5283 has been described as an inhibitor of phosphoinositide\u2010specific PLC [http://www.ncbi.nlm.nih.gov/pubmed/2338654?dopt=AbstractPlus], although its selectivity among the isoforms is untested and it has been reported to occupy the H1 histamine receptor [http://www.ncbi.nlm.nih.gov/pubmed/11138848?dopt=AbstractPlus].Phosphoinositide\u2010specific phospholipase C , catalyses the hydrolysis of https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:9063, http://www.uniprot.org/uniprot/Q15111; https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:9064, http://www.uniprot.org/uniprot/Q9UPR0 and https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:29185, http://www.uniprot.org/uniprot/Q4KWH8) form a family with PLC\u03b4 and PLC\u03b61 isoforms, but appear to lack catalytic activity. PLC\u2010\u03b42 has been cloned from bovine sources [http://www.ncbi.nlm.nih.gov/pubmed/1848183?dopt=AbstractPlus].A series of PLC\u2010like proteins Phosphoinositide\u2010specific phospholipase C in health and disease. J. Lipid Res.56: 1853\u201360 https://www.ncbi.nlm.nih.gov/pubmed/25821234?dopt=AbstractPlusCocco L et al. (2016) Topological organisation of the phosphatidylinositol 4,5\u2010bisphosphatephospholipase C resynthesis cycle: PITPs bridge the ER\u2010PM gap. Biochem. J. 473: 4289\u20134310 https://www.ncbi.nlm.nih.gov/pubmed/27888240?dopt=AbstractPlusCockcroft S Life Sci. 137: 116\u201324 https://www.ncbi.nlm.nih.gov/pubmed/26239437?dopt=AbstractPlusLitosch I. (2015) Regulating G protein activity by lipase\u2010independent functions of phospholipase C. et al. (2017) Regulation and physiological functions of mammalian phospholipase C. J. Biochem. 161: 315\u2013321 https://www.ncbi.nlm.nih.gov/pubmed/28130414?dopt=AbstractPlusNakamura Y et al. (2016) The sperm phospholipase C\u2010\u03b6 and Ca2+ signalling at fertilization in mammals. Biochem. Soc. Trans. 44: 267\u201372 https://www.ncbi.nlm.nih.gov/pubmed/26862214?dopt=AbstractPlusSwann K 2 cleaves the sn\u20102 fatty acid of phospholipids, primarily phosphatidylcholine, to generate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2508 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2391. Most commonly\u2010used inhibitors are either non\u2010selective within the family of phospholipase A2 enzymes or have activity against other eicosanoid\u2010metabolising enzymes.Phospholipase A2\u20101B, sPLA2\u20102A, sPLA2\u20102D, sPLA2\u20102E, sPLA2\u20102F, sPLA2\u20103, sPLA2\u201010 and sPLA2\u201012AsPLA2\u20104A, cPLA2\u20104B, cPLA2\u20104C, cPLA2\u20104D, cPLA2\u20104E and cPLA2\u20104FcPLA2\u2010G5, iPLA2\u2010G6, PLA2\u2010G7 and PAFAH2 (platelet\u2010activating factor acetylhydrolase 2)PLA2\u20102C suggests a lack of catalytic activity, while PLA2\u201012B appears to be catalytically inactive [http://www.ncbi.nlm.nih.gov/pubmed/14516201?dopt=AbstractPlus]. A further fragment has been identified with sequence similarities to Group II PLA2 members. Otoconin 90 (OC90) shows sequence homology to PLA2\u2010G10.The sequence of PLA2 has been identified which shows modest selectivity for sPLA2\u20101B over sPLA2\u20102A, and also binds snake toxin phospholipase A2 [http://www.ncbi.nlm.nih.gov/pubmed/7548076?dopt=AbstractPlus]. The binding protein appears to have clearance function for circulating secretory phospholipase A2, as well as signalling functions, and is a candidate antigen for idiopathic membraneous nephropathy [http://www.ncbi.nlm.nih.gov/pubmed/19571279?dopt=AbstractPlus].A binding protein for secretory phospholipase A2\u2010G7 and PAFAH2 also express platelet\u2010activating factor acetylhydrolase activity (http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=3.1.1.47).PLABiochim Biophys Acta Mol Cell Biol Lipids1864: 772\u2013783 https://www.ncbi.nlm.nih.gov/pubmed/30010011Astudillo AM. (2019) Selectivity of phospholipid hydrolysis by phospholipase A2 enzymes in activated cells leading to polyunsaturated fatty acid mobilization. etal. (2019) Cytosolic phospholipase A2 and lysophospholipid acyltransferases. Biochim Biophys Acta Mol Cell Biol Lipids1864: 838\u2013845 https://www.ncbi.nlm.nih.gov/pubmed/30905348Kita Y Biochim Biophys Acta Mol Cell Biol Lipids1864: 766\u2013771 https://www.ncbi.nlm.nih.gov/pubmed/30905345Mouchlis VD and Dennis EA. (2019) Phospholipase A2 catalysis and lipid. mediator lipidomics. et al. (2019) Group IID, IIE, IIF and III secreted phospholipase A2s. Biochim Biophys Acta Mol Cell Biol Lipids. 1864: 803\u2013818 https://www.ncbi.nlm.nih.gov/pubmed/30905347Murakami M Biochim Biophys Acta Mol Cell Biol Lipids1864: 819\u2013826 https://www.ncbi.nlm.nih.gov/pubmed/30308324Samuchiwal SK and Balestrieri B. (2019) Harmful and protective roles of group V phospholipase A2: Current perspectives and future directions. Biochim Biophys Acta Mol Cell Biol Lipids. 1864: 932\u2013940 https://www.ncbi.nlm.nih.gov/pubmed/30077006Shayman JA and Tesmer JJG. (2019) Lysosomal phospholipase A2. http://www.ncbi.nlm.nih.gov/pubmed/2186929?dopt=AbstractPlus].Phosphatidylcholine\u2010specific phospholipase D catalyses the formation of phosphatidic acid from phosphatidylcholine. In addition, the enzyme can make use of alcohols, such as butanol in a transphosphatidylation reaction . This enzyme activity appears to be enhanced by polyamines in the physiological range [http://www.ncbi.nlm.nih.gov/pubmed/12047899?dopt=AbstractPlus] and fails to transphosphatidylate with alcohols [http://www.ncbi.nlm.nih.gov/pubmed/10428468?dopt=AbstractPlus].A lysophospholipase D activity , PLD4 and PLD5 . PLD3 has been reported to be involved in myogenesis [http://www.ncbi.nlm.nih.gov/pubmed/22428023?dopt=AbstractPlus]. PLD4 is described not to have phospholipase D catalytic activity [http://www.ncbi.nlm.nih.gov/pubmed/21085684?dopt=AbstractPlus], but has been associated with inflammatory disorders . Sequence analysis suggests that PLD5 is catalytically inactive.Three further, less well\u2010characterised isoforms are PLD3 Targeting phospholipase D in cancer, infection and neurodegenerative disorders. Nat Rev Drug Discov16: 351\u2010367 https://www.ncbi.nlm.nih.gov/pubmed/28209987?dopt=AbstractPlusBrown HA Trends Pharmacol. Sci. 36: 137\u201044 https://www.ncbi.nlm.nih.gov/pubmed/25661257?dopt=AbstractPlusFrohman MA. (2015) The phospholipase D superfamily as therapeutic targets. et al. (2015) Physiological and pathophysiological roles for phospholipase D. J. Lipid Res. 56: 2229\u201037 https://www.ncbi.nlm.nih.gov/pubmed/25926691?dopt=AbstractPlusNelson RK Lipid phosphate phosphatases, divided into phosphatidic acid phosphatases or lipins catalyse the dephosphorylation of phosphatidic acid (and other phosphorylated lipid derivatives) to generate inorganic phosphate and diacylglycerol. PTEN, a phosphatase and tensin homolog is a phosphatidylinositol 3,4,5\u2010trisphosphate 3\u2010phosphatase which acts as a tumour suppressor by reducing cellular levels of PI 3,4,5\u2010P, thereby toning down activity of PDK1 and PKB. Loss\u2010of\u2010function mutations are frequently identified as somatic mutations in cancers.Trends Neurosci40: 83\u201091 https://www.ncbi.nlm.nih.gov/pubmed/28081942Knafo S and Esteban JA. (2017) PTEN: Local and Global Modulation of Neuronal Function in Health and Disease. et al. (2018) The functions and regulation of the PTEN tumour suppressor: new modes and prospects. Nat Rev Mol Cell Biol19: 547\u2010562 https://www.ncbi.nlm.nih.gov/pubmed/29858604Lee YR et al. (2019) PTEN\u2010opathies: from biological insights to evidence\u2010based precision medicine. J Clin Invest129: 452\u2010464 https://www.ncbi.nlm.nih.gov/pubmed/30614812Yehia L Phosphatidylinositol may be phosphorylated at either 3\u2010 or 4\u2010 positions on the inositol ring by PI 3\u2010kinases or PI 4\u2010kinases, respectively.2). There is evidence that PI3K can also phosphorylate serine/threonine residues on proteins. In addition to the classes described below, further serine/threonine protein kinases, including https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:795 (http://www.uniprot.org/uniprot/Q13315) and https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:3942 (http://www.uniprot.org/uniprot/P42345), have been described to phosphorylate phosphatidylinositol and have been termed PI3Krelated kinases. Structurally, PI3Ks have common motifs of at least one C2, calcium\u2010binding domain and helical domains, alongside structurally\u2010conserved catalytic domains. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6060 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6004 are widely\u2010used inhibitors of PI3K activities. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6060 is irreversible and shows modest selectivity between Class I and Class II PI3K, while LY294002 is reversible and selective for Class I compared to Class II PI3K.Phosphatidylinositol 3\u2010kinases catalyse the introduction of a phosphate into the 3\u2010position of phosphatidylinositol (PI), phosphatidylinositol 4\u2010phosphate (PIP) or phosphatidylinositol 4,5\u2010bisphosphate (PIPClass I PI3Ks (EC 2.7.1.153) phosphorylate phosphatidylinositol 4,5\u2010bisphosphate to generate phosphatidylinositol 3,4,5\u2010trisphosphate and are heterodimeric, matching catalytic and regulatory subunits. Class IA PI3Ks include p110\u03b1, p110\u00df and p110\u03b4 catalytic subunits, with predominantly p85 and p55 regulatory subunits. The single catalytic subunit that forms Class IB PI3K is p110\u03b3. Class IA PI3Ks are more associated with receptor tyrosine kinase pathways, while the Class IB PI3K is linkedmore with GPCR signalling.Class II PI3Ks (EC 2.7.1.154) phosphorylate phosphatidylinositol to generate phosphatidylinositol 3\u2010phosphate . Three monomeric members exist, PI3KC2\u03b1, \u00df and \u00df, and include Ras\u2010binding, Phox homology and two C2domains.class III PI3K isoform (EC 2.7.1.137) is a heterodimer formed of a catalytic subunit (VPS34) and regulatory subunit (VPS15).The only Phosphatidylinositol 4\u2010kinasesPhosphatidylinositol 4\u2010kinases (EC 2.7.1.67) generate phosphatidylinositol 4\u2010phosphate and may be divided into higher molecular weight type III and lower molecular weight type II forms.et al. (2017) [http://www.ncbi.nlm.nih.gov/pubmed/28802037?dopt=AbstractPlus].PI3K activation is one of the most important signal transduction pathways used to transmit signals from cell\u2010surface receptors to regulate intracellular processes . PI3K catalytic (and regulatory) subunits play vital roles in normal cell function and in disease. Progress made in developing PI3K\u2010targeted agents as potential therapeutics for treating cancer and other diseases is reviewed by Fruman http://www.ncbi.nlm.nih.gov/pubmed/9367159?dopt=AbstractPlus]. This enzyme family is also known as type I PIP(5)Ks.Type I PIP kinases are required for the production of the second messenger phosphatidylinositol 4,5\u2010bisphosphate P2) by phosphorylating PtdIns(4)P [http://www.ncbi.nlm.nih.gov/pubmed/9367159?dopt=AbstractPlus]. This enzyme family is also known as type II PIP(5)Ks.Type II PIP kinases are essential for the production of the second messenger phosphatidylinositol 4,5\u2010bisphosphate P2) by phosphorylating PtdIns(5)P ) and induces proteasomal degradation of SK1 [http://www.ncbi.nlm.nih.gov/pubmed/26934645?dopt=AbstractPlus]. ABC294640 is in clinical trials for advanced cholangiocarcinoma, advanced hepatocellular carcinoma and refractory/relapsed multiple myeloma .et al. (2016) Sphingosine Kinases: Emerging Structure\u2010Function Insights. Trends Biochem. Sci. 41: 395\u2010409 https://www.ncbi.nlm.nih.gov/pubmed/27021309?dopt=AbstractPlusAdams DR et al. (2016) Sphingosine kinase inhibitors: a review of patent literature (2006\u20102015). Expert Opin Ther Pat26: 1409\u20101416 https://www.ncbi.nlm.nih.gov/pubmed/27539678?dopt=AbstractPlusLynch KR et al. (2016) Recent advances in the development of sphingosine kinase inhibitors. Cell. Signal. 28: 1349\u201063 https://www.ncbi.nlm.nih.gov/pubmed/27297359?dopt=AbstractPlusPitman MR et al. (2018) An intrinsic lipid\u2010binding interface controls sphingosine kinase 1 function. J. Lipid Res. 59: 462\u2010474 https://www.ncbi.nlm.nih.gov/pubmed/29326159?dopt=AbstractPlusPulkoski\u2010Gross MJ et al. (2017) Sphingosine Kinase 2 in Autoimmune/Inflammatory Disease and the Development of Sphingosine Kinase 2 Inhibitors. Trends Pharmacol. Sci. 38: 581\u2010591 https://www.ncbi.nlm.nih.gov/pubmed/28606480?dopt=AbstractPlusPyne NJ et al. (2018) Sphingosine Kinases as Druggable Targets. Handb Exp Pharmacolhttps://www.ncbi.nlm.nih.gov/pubmed/29460151?dopt=AbstractPlusPyne S http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6060 also inhibits type III phosphatidylinositol 4\u2010kinases and polo\u2010like kinase [http://www.ncbi.nlm.nih.gov/pubmed/15664519?dopt=AbstractPlus]. PIK93 also inhibits PI 3\u2010kinases [http://www.ncbi.nlm.nih.gov/pubmed/16647110?dopt=AbstractPlus]. Adenosine activates http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=3.et al. (2018) Phosphoinositide 3\u2010kinase inhibitors in advanced breast cancer: A systematic review and meta\u2010analysis. Eur J Cancer91: 38\u201046 https://www.ncbi.nlm.nih.gov/pubmed/29331750Raphael J et al. (2019) Upstream regulators of phosphoinositide 3\u2010kinase and their role in diseases. J Cell Physiol. https://www.ncbi.nlm.nih.gov/pubmed/30710358Wang D et al. (2018) Phosphatidylinositol 3\u2010Kinase, Growth Disorders, and Cancer. N Engl J Med379: 2052\u20102062 https://www.ncbi.nlm.nih.gov/pubmed/30462943Goncalves MD 2 is generated by phosphorylation of PI 4\u2010phosphate or PI 5\u2010phosphate by type I PI 4\u2010phosphate 5\u2010kinases or type II PI 5\u2010phosphate 4\u2010kinases.PIPet al. (2015) Phosphoinositides: Lipids with informative heads and mastermind functions in cell division. Biochim. Biophys. Acta1851: 832\u201043 https://www.ncbi.nlm.nih.gov/pubmed/25449648?dopt=AbstractPlusCauvin C J. Lipid Res. 57: 1987\u20101994 https://www.ncbi.nlm.nih.gov/pubmed/27623846?dopt=AbstractPlusIrvine RF. (2016) A short history of inositol lipids. et al. (2016) Nuclear Phosphatidylinositol Signaling: Focus on Phosphatidylinositol Phosphate Kinases and Phospholipases C. J. Cell. Physiol. 231: 1645\u201055 https://www.ncbi.nlm.nih.gov/pubmed/26626942?dopt=AbstractPlusPoli A http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=1.14.99.3, converts http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4349 into http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5153 and carbonmonoxide, utilizing http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3041 as cofactor.Haem oxygenase ), http://www.ncbi.nlm.nih.gov/pubmed/15246535?dopt=AbstractPlus]. The chemical http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5279 acts as a haem oxygenase inhibitor in rat liver with an IC50 value of 11 nM [http://www.ncbi.nlm.nih.gov/pubmed/6947237?dopt=AbstractPlus].The existence of a third non\u2010catalytic version of haem oxygenase, HO3, has been proposed, although this has been suggested to be a pseudogene .Hydrogen sulfide is a gasotransmitter, with similarities to nitric oxide and carbon monoxide. Although the enzymes indicated below have multiple enzymatic activities, the focus here is the generation of hydrogen sulphide Selectivity of commonly used pharmacological inhibitors for cystathionine \u03b2 synthase (CBS) and cystathionine \u03b3 lyase (CSE). Br J Pharmacol. 169: 922\u201032 https://www.ncbi.nlm.nih.gov/pubmed/23488457?dopt=AbstractPlusAsimakopoulou A et al. (2017) International Union of Basic and Clinical Pharmacology. CII: Pharmacological Modulation of H2S Levels: H2S Donors and H2S Biosynthesis Inhibitors. Pharmacol. Rev. 69: 497\u2010564 https://www.ncbi.nlm.nih.gov/pubmed/28978633?dopt=AbstractPlusSzabo C http://www.uniprot.org/uniprot/P23141), but predominates in the liver, where it is responsible for the hydrolysis of many aliphatic, aromatic and steroid esters. Hormone\u2010sensitive lipase is also a relatively non\u2010selective esterase associated with steroid ester hydrolysis and triglyceride metabolism, particularly in adipose tissue. Endothelial lipase is secreted from endothelial cells and regulates circulating cholesterol in high density lipoproteins.Listed in this section are hydrolases not accumulated in other parts of the Concise Guide, such as monoacylglycerol lipase and acetylcholinesterase. Pancreatic lipase is the predominant mechanism of fat digestion in the alimentary system; its inhibition is associated with decreased fat absorption. CES1 is present at lower levels in the gut than CES2 The ectonucleotidases CD39 and CD73: Novel checkpoint inhibitor targets. Immunol Rev. 276: 121\u2010144 https://www.ncbi.nlm.nih.gov/pubmed/28258700Allard B et al. (2018) CD39\u2010adenosinergic axis in renal pathophysiology and therapeutics. Purinergic Signal14: 109\u2010120 https://www.ncbi.nlm.nih.gov/pubmed/29332180Kishore BK et al. (2018) Carboxylesterase 1 genes: systematic review and evaluation of existing genotyping procedures. Drug Metab Pers Ther33: 3\u201014 https://www.ncbi.nlm.nih.gov/pubmed/29427553Rasmussen HB et al. (2018) Carboxylesterase Inhibitors: An Update. Curr Med Chem. 25: 1627\u20101649 https://www.ncbi.nlm.nih.gov/pubmed/29210644Zou LW myo\u2010inositol is a component of the http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=244, where the principal second messenger is inositol 1,4,5\u2010trisphosphate, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4222, which acts at intracellular ligand\u2010gated ion channels, http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=123 to elevate intracellular calcium. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4222 is recycled to inositol by phosphatases or phosphorylated to form other active inositol polyphosphates. Inositol produced from dephosphorylation of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4222 is recycled into membrane phospholipid under the influence of phosphatidyinositol synthase activity (CDP\u2010diacylglycerol\u2010inositol 3\u2010phosphatidyltransferase [http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=2.7.8.11]).The sugar alcohol D\u2010http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=2.7.1.127, http://www.ensembl.org/Homo_sapiens/Gene/Family/Genes?family=ENSFM00250000001260) catalyse the generation of inositol 1,3,4,5\u2010tetrakisphosphate (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5202) from http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4222. IP3 kinase activity is enhanced in the presence of calcium/http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2351 [http://www.ncbi.nlm.nih.gov/pubmed/2559811?dopt=AbstractPlus].Inositol 1,4,5\u2010trisphosphate 3\u2010kinases generates http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5119, 4\u2010phosphatases generate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5099 and 5\u2010phosphatases (http://www.genome.jp/dbget\u2010bin/www_bget?ec:3.1.3.36 or http://www.genome.jp/dbget\u2010bin/www_bget?ec:3.1.3.56) generate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5098.Members of this family exhibit phosphatase activity towards IPhttp://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=281.Information on members of this family may be found in the In vitro analysis suggested http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4222 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5202 were poor substrates for SKIP, synaptojanin 1 and synaptojanin 2, but suggested that http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2387 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2353 were more efficiently hydrolysed [http://www.ncbi.nlm.nih.gov/pubmed/15474001?dopt=AbstractPlus].http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=3.1.3.25, IMPase, myo\u2010inositol\u20101(or 4)\u2010phosphate phosphohydrolase) is a magnesium\u2010dependent homodimer which hydrolyses myoinositol monophosphate to generate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4495 and phosphate. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5195 may be a physiological phosphate acceptor. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5212 is a nonselective un\u2010competitive inhibitor more potent at IMPase 1 than IMPase 2 . IMPase activity may be inhibited competitively by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5208 , although the enzyme selectivity is not yet established.Inositol monophosphatase A tale of two inositol trisphosphates. et al. (2016) Phosphate, inositol and polyphosphates. Biochem. Soc. Trans. 44: 253\u20109 https://www.ncbi.nlm.nih.gov/pubmed/26862212?dopt=AbstractPlusLivermore TM et al. (2017) Probes for manipulating and monitoring IP_3. Cell Calcium64: 57\u201064 https://www.ncbi.nlm.nih.gov/pubmed/27887748?dopt=AbstractPlusMiyamoto A et al. (2017) Inositol\u20101,4,5\u2010trisphosphate 3\u2010kinase\u2010A (ITPKA) is frequently over\u2010expressed and functions as an oncogene in several tumor types. Biochem. Pharmacol. 137: 1\u20109 https://www.ncbi.nlm.nih.gov/pubmed/28377279?dopt=AbstractPlusWindhorst S http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=2.7.11.1) use the co\u2010substrate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1713 to phosphorylate serine and/or threonine residues on target proteins. Analysis of the human genome suggests the presence of 518 protein kinases in man (divided into 15 subfamilies), with over 100 protein kinase\u2010like pseudogenes [http://www.ncbi.nlm.nih.gov/pubmed/12471243?dopt=AbstractPlus]. It is beyond the scope of the Concise Guide to list all these protein kinase activities, but full listings are available on the \u2019Detailed page\u2019 provided for each enzyme.Protein kinases , which in turn may be activated by G\u03b112/13 subunits [http://www.ncbi.nlm.nih.gov/pubmed/9641915?dopt=AbstractPlus].Rho kinase is activated by members of the Rho small G protein family, which are activated by GTP exchange factors, such as et al. (2016) Rho Kinase (ROCK) Inhibitors and Their Therapeutic Potential. J. Med. Chem. 59: 2269\u2010300 https://www.ncbi.nlm.nih.gov/pubmed/26486225?dopt=AbstractPlusFeng Y et al. (2015) Developing novel methods to search for substrates of protein kinases such as Rho\u2010kinase. Biochim. Biophys. Acta1854: 1663\u20106 https://www.ncbi.nlm.nih.gov/pubmed/25770685?dopt=AbstractPlusNishioka T et al. (2016) RhoA/Rho\u2010Kinase in the Cardiovascular System. Circ. Res. 118: 352\u201066 https://www.ncbi.nlm.nih.gov/pubmed/26838319?dopt=AbstractPlusShimokawa H http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2341).Protein kinase C is the target for the tumour\u2010promoting phorbol esters, such as tetradecanoyl\u2010\u03b2\u2010phorbol acetate Dynamics and Membrane Interactions of Protein Kinase C. et al. (2017) Reversing the Paradigm: Protein Kinase C as a Tumor Suppressor. Trends Pharmacol. Sci. 38: 438\u2010447 https://www.ncbi.nlm.nih.gov/pubmed/28283201?dopt=AbstractPlusNewton AC et al. (2016) Protein Kinase C \u03b4: a Gatekeeper of Immune Homeostasis. J. Clin. Immunol. 36: 631\u201040 https://www.ncbi.nlm.nih.gov/pubmed/27541826?dopt=AbstractPlusSalzer E et al. (2016) The cytotoxic T cell proteome and its shaping by the kinase mTOR. Nat. Immunol. 17: 104\u201012 https://www.ncbi.nlm.nih.gov/pubmed/26551880?dopt=AbstractPlusHukelmann JL et al. (2017) mTOR Signaling in Growth, Metabolism, and Disease. Cell169: 361\u2010371 https://www.ncbi.nlm.nih.gov/pubmed/28388417?dopt=AbstractPlusSaxton RA http://www.ncbi.nlm.nih.gov/pubmed/24879308?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/22512864?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/25561469?dopt=AbstractPlus].Five of the cyclin\u2010dependent kinases are involved in the phosphorylation of serine residues in the C\u2010terminal domain of RNA polymerase II, the enzyme that is responsible for the transcription of protein\u2010coding genes into mRNA in eukaryotes. Phosphorylation of RNA polymerase II at Ser5 is essential for transcriptional initiation, and phosphorylation of Ser 2 contributes to transcriptional elongation and termination. All five of the C\u2010terminal domain kinases can phosphorylate Ser5, but only CDK9, CDK12, and CDK13 can phosphorylate at Ser2 [http://www.ncbi.nlm.nih.gov/pubmed/26115571?dopt=AbstractPlus], with detailed content covering CDK4 and CDK6 inhibitors that are under clinical evaluation. Data produced by Jorda et al. (2018) highlights the caution that must be used when deploying commercially available CDK inhibitors as pharmacological probes [http://www.ncbi.nlm.nih.gov/pubmed/30234987?dopt=AbstractPlus], as most of them are more promiscuous in their selectivity than indicated. To make their findings easily accessible the Jorda data is hosted on the http://rustreg.upol.cz/CDKiDB/.The development of CDK inhibitors as anticancer drugs is reviewed in .Since it is beyond the scope of the Guide to list all peptidase and proteinase activities, this summary focuses on selected enzymes of significant pharmacological interest that have ligands directed against them. For those interested in detailed background we recommend the MEROPS database (with whhttp://www.ncbi.nlm.nih.gov/pubmed/2881207?dopt=AbstractPlus] in the generation of amyloid beta (A\u03b2) . Given that the accumulation and aggregation of A\u03b2 in the brain is pivotal in the development of Alzheimer's disease (AD), inhibition of PS activity is one mechanism being investigated as a therapeutic option for AD [http://www.ncbi.nlm.nih.gov/pubmed/11378516?dopt=AbstractPlus]. Several small molecule inhibitors of PS\u20101 have been investigated, with some reaching early clinical trials, but none have been formally approved. Dewji et al. (2015) have reported that small peptide fragments of human PS\u20101 can significantly inhibit A\u03b2 production both in vitro and when infused in to the brains of APP transgenic mice [http://www.ncbi.nlm.nih.gov/pubmed/25923432?dopt=AbstractPlus]. The most active small peptides in this report were http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=8344 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=8345, from the amino\u2010terminal domain of PS\u20101.Presenilin (PS)\u20101 or \u20102 act as the catalytic component/ essential co\u2010factor of the \u03b3\u2010secretase complex responsible for the final carboxy\u2010terminal cleavage of amyloid precursor protein (APP) ) on functional and structural bases into gelatinases, collagenases, stromyelinases and matrilysins, as well as membrane type\u2010MMP (MT\u2010MMP).Matrix metalloproteinases (MMP) are calcium\u2010 and zinc\u2010dependent proteinases regulating the extracellular matrix and are often divided proteins are endogenous inhibitors acting to chelate MMP proteins: http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5309 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5310 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5311 , http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5312 ADAM metalloproteinases cleave cell\u2010surface or transmembrane proteins to generate soluble and membrane\u2010limited products.ADAMTS (with thrombospondin motifs) metalloproteinases cleave cell\u2010surface or transmembrane proteins to generate soluble and membrane\u2010limited products.http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=739.Information on members of this family may be found in the http://www.ensembl.org/Homo_sapiens/Gene/Summary?g=ENSG00000235812;r=14:70712470\u201070714518), AC136428.3\u20102 (ENSG00000185520) and ADAMDEC1 .Additional ADAM family members include AC123767.2 , AL160191.3 , AC139425.3\u20101 (ENSG00000225577), and AC126339.6\u20101 (ENSG00000225734).http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6878 has been used for imaging purposes.Folate hydrolase is also known as NAALADase as it is responsible for the hydrolysis of N\u2010acetaspartylglutamate to form N\u2010acetylaspartate and L\u2010glutamate. In the gut, the enzyme assists in the assimilation of folate by hydrolysing dietary poly\u2010gamma\u2010glutamylfolate. The enzyme is highly expressed in the prostate, and its expression is up\u2010regulated in cancerous tissue. A tagged version of the antibody http://www.ncbi.nlm.nih.gov/pubmed/16142822?dopt=AbstractPlus]. This catalytic core enables the degradation of peptides with Arg, Phe, Tyr, Leu, and Glu adjacent to the cleavage site. The \u03b25 subunit is the principal target of the approved drug proteasome inhibitor http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6391.The T1 macropain beta subunits form the catalytic proteinase core of the 20S proteasome complex . Therapeutics which inhibit PCSK9 are viewed as potentially lucrative replacements for statins, upon statin patent expiry. Several monoclonal antibodies including http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6744, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7343, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7730, RG\u20107652 and LY3015014 are under development. One RNAi therapeutic, code named ALN\u2010PCS02, is also in development .One member of this family has garnered intense interest as a clinical drug target. As liver PCSK9 acts to maintain cholesterol homeostasis, it has become a target of intense interest for clinical drug development. Inhibition of PCSK9 can lower low\u2010density cholesterol (LDL\u2010C) by clearing LDLR\u2010bound LDL particles, thereby lowering circulating cholesterol levels. It is hypothesised that this action may improve outcomes in patients with atherosclerotic cardiovascular disease ) and induces proteasomal degradation of SK1 [http://www.ncbi.nlm.nih.gov/pubmed/26934645?dopt=AbstractPlus]. ABC294640 is in clinical trials for advanced cholangiocarcinoma, advanced hepatocellular carcinoma and refractory/relapsed multiple myeloma .et al. (2016) Sphingosine Kinases: Emerging Structure\u2010Function Insights. Trends Biochem. Sci. 41: 395\u2010409 https://www.ncbi.nlm.nih.gov/pubmed/27021309?dopt=AbstractPlusAdams DR et al. (2016) Sphingosine kinase inhibitors: a review of patent literature (2006\u20102015). Expert Opin Ther Pat26: 1409\u20101416 https://www.ncbi.nlm.nih.gov/pubmed/27539678?dopt=AbstractPlusLynch KR et al. (2016) Recent advances in the development of sphingosine kinase inhibitors. Cell. Signal. 28: 1349\u201063 https://www.ncbi.nlm.nih.gov/pubmed/27297359?dopt=AbstractPlusPitman MR et al. (2018) An intrinsic lipid\u2010binding interface controls sphingosine kinase 1 function. J. Lipid Res. 59: 462\u2010474 https://www.ncbi.nlm.nih.gov/pubmed/29326159?dopt=AbstractPlusPulkoski\u2010Gross MJ et al. (2017) Sphingosine Kinase 2 in Autoimmune/Inflammatory Disease and the Development of Sphingosine Kinase 2 Inhibitors. Trends Pharmacol. Sci. 38: 581\u2010591 https://www.ncbi.nlm.nih.gov/pubmed/28606480?dopt=AbstractPlusPyne NJ et al. (2018) Sphingosine Kinases as Druggable Targets. Handb Exp Pharmacolhttps://www.ncbi.nlm.nih.gov/pubmed/29460151?dopt=AbstractPlusPyne S http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=911. The phenotype of Sgpp1(\u2010/\u2010) mice differ with genetic background. Sgpp2(\u2010/\u2010) mice are also available. No specific SGPP inhibitors available [http://www.ncbi.nlm.nih.gov/pubmed/22052905?dopt=AbstractPlus].SGPP1 and SGPP2 are non\u2010redundant endoplasmic reticulum enzymes that dephosphorylate intracellular et al. (2013) Sphingosine\u20101\u2010phosphate phosphatase 1 regulates keratinocyte differentiation and epidermal homeostasis. J. Biol. Chem. 288: 18381\u201091 https://www.ncbi.nlm.nih.gov/pubmed/23637227?dopt=AbstractPlusAllende ML et al. (2016) Sphingosine\u20101\u2010phosphate phosphatase 2 promotes disruption of mucosal integrity, and contributes to ulcerative colitis in mice and humans. FASEB J. 30: 2945\u201058 https://www.ncbi.nlm.nih.gov/pubmed/27130484?dopt=AbstractPlusHuang WC et al. (2011) Sphingosine\u20101\u2010phosphate phosphohydrolase\u20101 regulates ER stress\u2010induced autophagy. Cell Death Differ. 18: 350\u201061 https://www.ncbi.nlm.nih.gov/pubmed/20798685?dopt=AbstractPlusL\u00e9pine S et al. (2000) Molecular cloning and characterization of a lipid phosphohydrolase that degrades sphingosine\u20101\u2010 phosphate and induces cell death. Proc. Natl. Acad. Sci. U.S.A. 97: 7859\u201064 https://www.ncbi.nlm.nih.gov/pubmed/10859351?dopt=AbstractPlusMandala SM et al. (2016) Sphingosine\u20101\u2010phosphate Phosphatase 2 Regulates Pancreatic Islet \u03b2\u2010Cell Endoplasmic Reticulum Stress and Proliferation. J. Biol. Chem. 291: 12029\u201038 https://www.ncbi.nlm.nih.gov/pubmed/27059959?dopt=AbstractPlusTaguchi Y http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6626 (2\u2010Acetyl\u20105\u2010tetrahydroxybutyl imidazole) inhibits the enzyme activity in intact cell preparations [http://www.ncbi.nlm.nih.gov/pubmed/16151014?dopt=AbstractPlus]. Recessive mutations in the S1P lyase (SGPL1) gene underlie a recently identified sphingolipidosis: SPL Insufficiency Syndrome (SPLIS) [http://www.ncbi.nlm.nih.gov/pubmed/30274713?dopt=AbstractPlus]. A Phase 2 clinical trial of LX3305 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=9851) for rheumatoid arthritis has been completed .et al. (2018) A novel mutation in sphingosine\u20101\u2010phosphate lyase causing congenital brain malformation. Brain Dev. 40: 480\u2010483 https://www.ncbi.nlm.nih.gov/pubmed/29501407?dopt=AbstractPlusBamborschke D et al. (2019) Sphingosine phosphate lyase insufficiency syndrome (SPLIS): A novel inborn error of sphingolipid metabolism. Adv Biol Regul71: 128\u2010140 https://www.ncbi.nlm.nih.gov/pubmed/30274713Choi YJ et al. (2017) Mutations in sphingosine\u20101\u2010phosphate lyase cause nephrosis with ichthyosis and adrenal insufficiency. J. Clin. Invest. 127: 912\u2010928 https://www.ncbi.nlm.nih.gov/pubmed/28165339?dopt=AbstractPlusLovric S et al. (2017) Sphingosine\u20101\u2010phosphate lyase mutations cause primary adrenal insufficiency and steroid\u2010resistant nephrotic syndrome. J. Clin. Invest. 127: 942\u2010953 https://www.ncbi.nlm.nih.gov/pubmed/28165343?dopt=AbstractPlusPrasad R http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2634 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2635, respectively, are synthesized in the thyroid gland by sequential metabolism of tyrosine residues in the glycosylated homodimeric protein thyroglobulin under the influence of the haem\u2010containing protein iodide peroxidase. Iodide peroxidase/TPO is a haem\u2010containing enzyme, from the same structural family as eosinophil peroxidase , lactoperoxidase and myeloperoxidase . Circulating thyroid hormone is bound to thyroxine\u2010binding globulin .The thyroid hormones triiodothyronine and thyroxine, usually abbreviated as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2635 to generate http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2634 or http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2636 . DIO1 is also able to deiodinate RT3 to form 3,3\u2032\u2010diidothyronine (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6648). Iodotyrosine deiodinase is a 1TM homodimeric enzyme.These are 1 TM selenoproteins that remove an iodine from et al. (2015) Intracellular thyroid hormone metabolism as a local regulator of nuclear thyroid hormone receptor\u2010mediated impact on vertebrate development. Biochim. Biophys. Acta1849: 130\u201041 https://www.ncbi.nlm.nih.gov/pubmed/24844179?dopt=AbstractPlusDarras VM et al. (2015) Scope and limitations of iodothyronine deiodinases in hypothyroidism. Nat Rev Endocrinol11: 642\u2010652 https://www.ncbi.nlm.nih.gov/pubmed/26416219?dopt=AbstractPlusGereben B et al. (2017) Novel thyroid hormone analogues, enzyme inhibitors andmimetics, and their action. Mol. Cell. Endocrinol. 458: 91\u2010104 https://www.ncbi.nlm.nih.gov/pubmed/28408161?dopt=AbstractPlusMondal S et al. (2015) New insights into the structure and mechanism of iodothyronine deiodinases. J. Mol. Endocrinol. 55: R37\u201052 https://www.ncbi.nlm.nih.gov/pubmed/26390881?dopt=AbstractPlusSchweizer U et al. (2017) Thyroid hormone metabolism in innate immune cells. J. Endocrinol. 232: R67\u2010R81 https://www.ncbi.nlm.nih.gov/pubmed/27852725?dopt=AbstractPlusvan der Spek AH et al. (2015) Challenges and Opportunities in the Discovery of New Therapeutics Targeting the Kynurenine Pathway. J. Med. Chem. 58: 8762\u201082 https://www.ncbi.nlm.nih.gov/pubmed/26207924?dopt=AbstractPlusDounay AB et al. (2017) The kynurenine pathway in schizophrenia and bipolar disorder. Neuropharmacology112: 297\u2010306 https://www.ncbi.nlm.nih.gov/pubmed/27245499?dopt=AbstractPlusErhardt S et al. (2017) L\u2010Tryptophan\u2010kynurenine pathway enzymes are therapeutic target for neuropsychiatric diseases: Focus on cell type differences. Neuropharmacology112: 264\u2010274 https://www.ncbi.nlm.nih.gov/pubmed/26767951?dopt=AbstractPlusFujigaki H et al. (2016) Kynurenine\u20103\u2010monooxygenase: a review of structure, mechanism, and inhibitors. Drug Discov. Today21: 315\u201024 https://www.ncbi.nlm.nih.gov/pubmed/26589832?dopt=AbstractPlusSmith JR et al. (2017) Abnormal kynurenine pathway of tryptophan catabolism in cardiovascular diseases. Cell. Mol. Life Sci. 74: 2899\u20102916 https://www.ncbi.nlm.nih.gov/pubmed/28314892?dopt=AbstractPlusSong P http://www.ncbi.nlm.nih.gov/pubmed/8621375?dopt=AbstractPlus]. Protein farnesyltransferase catalyses the post\u2010translational formation of a thioether linkage between the C\u20101 of an isoprenyl group and a cysteine residue fourth from the C\u2010terminus of a protein [http://www.ncbi.nlm.nih.gov/pubmed/7756316?dopt=AbstractPlus]. Farnesyltransferase is a dimer, composed of an alpha and beta subunit and requires Mg2+ and Zn2+ ions as cofactors. The active site is located between the subunits. Prenylation creates a hydrophobic domain on protein tails which acts as a membrane anchor.Farnesyltransferase is a member of the prenyltransferases family which also includes geranylgeranyltransferase types I (EC 2.5.1.59) and II (EC 2.5.1.60) .Classes I, II and IV use Znhttp://www.ncbi.nlm.nih.gov/pubmed/19608861?dopt=AbstractPlus] such as microtubules [http://www.ncbi.nlm.nih.gov/pubmed/12024216?dopt=AbstractPlus], the hsp90 chaperone [http://www.ncbi.nlm.nih.gov/pubmed/15916966?dopt=AbstractPlus] and the tumour suppressor p53 [http://www.ncbi.nlm.nih.gov/pubmed/11099047?dopt=AbstractPlus].HDACs have more general protein deacetylase activity, being able to deacetylate lysine residues in non\u2010histone proteins , making HDACs attractive molecular targets in the search for novel mechanisms to treat cancer [http://www.ncbi.nlm.nih.gov/pubmed/24382387?dopt=AbstractPlus]. Several small molecule HDAC inhibitors are already approved for clinical use: http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7006, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7496, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6852, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7489, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7496, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=7009 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=8305. HDACs and HDAC inhibitors currently in development as potential anti\u2010cancer therapeutics are reviewed by Sim\u00f3\u2010Riudalbas and Esteller (2015) [http://www.ncbi.nlm.nih.gov/pubmed/25039449?dopt=AbstractPlus].Dysregulated HDACactivity has been identified in cancer cells and tumour tissues . Overexpression and/or increased PADI activity is observed in several diseases, including rheumatoid arthritis, Alzheimer's disease, multiple sclerosis, lupus, Parkinson's disease, and cancer [http://www.ncbi.nlm.nih.gov/pubmed/23175390?dopt=AbstractPlus]. Pharmacological PADI inhibition reverses protein\u2010hypercitrullination and disease in mouse models of multiple sclerosis [http://www.ncbi.nlm.nih.gov/pubmed/23118341?dopt=AbstractPlus].In humans, the peptidyl arginine deiminases PAD4: pathophysiology, current therapeutics and future perspective in rheumatoid arthritis. Expert Opin. Ther. Targets21: 433\u2010447 https://www.ncbi.nlm.nih.gov/pubmed/28281906?dopt=AbstractPlusKoushik S et al. (2016) Peptidyl Arginine Deiminases and Neurodegenerative Diseases. Curr. Med. Chem. 23: 104\u201014 https://www.ncbi.nlm.nih.gov/pubmed/26577926?dopt=AbstractPlusTu R Neurochem. Int. 67: 23\u201031 https://www.ncbi.nlm.nih.gov/pubmed/24508404?dopt=AbstractPlusWhiteley CG. (2014) Arginine metabolising enzymes as targets against Alzheimers\u2019 disease. small G\u2010proteins, are a family of hydrolase enzymes that can bind and hydrolyze guanosine triphosphate (GTP). They are a type of G\u2010protein found in the cytosol that are homologous to the alpha subunit of heterotrimeric G\u2010proteins, but unlike the alpha subunit of G proteins, a small GTPase can function independently as a hydrolase enzyme to bind to and hydrolyze a guanosine triphosphate (GTP) to form guanosine diphosphate (GDP). The best\u2010known members are the Ras GTPases and hence they are sometimes called Ras subfamily GTPases.The RAS proteins are small membrane\u2010localised G protein\u2010likemolecules of 21 kd. They act as an on/off switch linking receptor and non\u2010receptor tyrosine kinase activation to downstream cytoplasmic or nuclear events. Binding of GTP activates the switch, and hydrolysis of the GTP to GDP inactivates the switch.http://www.ncbi.nlm.nih.gov/pubmed/7900159?dopt=AbstractPlus], which leads to increased cell proliferation and decreased apoptosis [http://www.ncbi.nlm.nih.gov/pubmed/17721087?dopt=AbstractPlus]. Because of their importance in oncogenic transformation these proteins have become the targets of intense drug discovery effort [http://www.ncbi.nlm.nih.gov/pubmed/22004085?dopt=AbstractPlus].The RAS proto\u2010oncogenes are the most frequently mutated class of proteins in human cancers. Common mutations compromise the GTP\u2010hydrolysing ability of the proteins causing constitutive activation [http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=897.Information on members of this family may be found in the et al. (2017) Deciphering the RAS/ERK pathway in vivo. Biochem. Soc. Trans. 45: 27\u201036 https://www.ncbi.nlm.nih.gov/pubmed/28202657?dopt=AbstractPlusDorard C et al. (2017) The RAS\u2010Effector Interaction as a Drug Target. Cancer Res. 77: 221\u2010226 https://www.ncbi.nlm.nih.gov/pubmed/28062402?dopt=AbstractPlusKeeton AB et al. (2016) Ras Conformational Ensembles, Allostery, and Signaling. Chem. Rev. 116: 6607\u201065 https://www.ncbi.nlm.nih.gov/pubmed/26815308?dopt=AbstractPlusLu S et al. (2016) Direct small\u2010molecule inhibitors of KRAS: from structural insights to mechanism\u2010based design. Nat Rev Drug Discov15: 771\u2010785 https://www.ncbi.nlm.nih.gov/pubmed/27469033?dopt=AbstractPlusOstrem JM et al. (2017) Drugging RAS: Know the enemy. Science355: 1158\u20101163 https://www.ncbi.nlm.nih.gov/pubmed/28302824?dopt=AbstractPlusPapke B et al. (2016) Pharmacological modulation of oncogenic Ras by natural products and their derivatives: Renewed hope in the discovery of novel anti\u2010Ras drugs. Pharmacol. Ther. 162: 35\u201057 https://www.ncbi.nlm.nih.gov/pubmed/27016467?dopt=AbstractPlusQuah SY et al. (2017) RAS Proteins and Their Regulators in Human Disease. Cell170: 17\u201033 https://www.ncbi.nlm.nih.gov/pubmed/28666118?dopt=AbstractPlusSimanshu DK http://www.genenames.org/cgi\u2010bin/genefamilies/set/388, http://www.genenames.org/cgi\u2010bin/genefamilies/set/388 ).The Rab family of proteins is a member of the Ras superfamily of monomeric G proteins. Rab GTPases regulate many steps of membrane traffic, including vesicle formation, vesicle movement along actin and tubulin networks, and membrane fusion. These processes make up the route through which cell surface proteins are trafficked from the Golgi to the plasma membrane and are recycled. Surface protein recycling returns proteins to the surface whose function involves carrying another protein or substance inside the cell, such as the transferrin receptor, or serves as a means of regulating the number of a certain type of protein molecules on the surface ( see http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=938.Information on members of this family may be found in the"} +{"text": "NORDLAND study. PLoS ONE 14(6): e0218521. https://doi.org/10.1371/journal.pone.0218521The third author\u2019s name is incorrect. The correct name is: Knut-Tore Lappegard. The correct citation is: Catalan-Matamoros D, Lopez-Villegas A, Tore-Lappegard K, Lopez-Liria R (2019) Patients' experiences of remote communication after pacemaker implant: The"} +{"text": "This article has been corrected: The correct author affiliation information is given below:1,2Hong Zhao84102-84111. https://doi.org/10.18632/oncotarget.21106Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: The authors recently became aware that the antibody used for 5545-5561. https://doi.org/10.18632/oncotarget.23798 Original article: Oncotarget. 2018; 9:5545\u20135561."} +{"text": "This article has been corrected: The correct author name is given below:Sepideh Saadatmand24335-24346. https://doi.org/10.18632/oncotarget.25262Original article: Oncotarget. 2018; 9:"} +{"text": "Correction to: Mol Med 22:224\u2013232, 2016DOI 10.2119/molmed.2016.00003Following publication of the original article (Sonkar et al."} +{"text": "This article has been corrected: The corrected author name is given below:Christina Gavegnano94040-94053. https://doi.org/10.18632/oncotarget.21541Original article: Oncotarget. 2017; 8:"} +{"text": "Correction to: BMC Health Services Research (2018) 18:575https://doi.org/10.1186/s12913-018-3368-3Following publication of the original article , the autTable Table Incorrect values:1.088(0.621\u20131.906)Single vs. Non-Users:1.067(0.721\u20131.581)Multiple vs. Single:Correct values:1.061(0.721\u20131.563)Single vs. Non-Users:1.047(0.815\u20131.345)Multiple vs. Single:"} +{"text": "There are errors in the Author Contributions. The correct contributions are: Conceptualization: HZ RWM FGG. Data curation: HZ RG KM FGG. Formal analysis: HZ RG MW FGG. Funding acquisition: HZ YPZ. Investigation: HZ MW FGG. Methodology: HZ MW FGG. Project administration: HZ YPZ. Resources: HZ FGG. Software: HZ FGG. Supervision: HZ RWM YPZ FGG. Validation: HZ RWM JCA RG PRMF GGM ABQ NLO MW YPZ FGG. Visualization: HZ JCA RG PRMF KM GGM ABQ NLO MW YPZ FGG. Writing\u2013original draft: HZ JCA RG PRMF GGM ABQ MW FGG. Writing\u2013review & editing: HZ RWM JCA RG PRMF KM GGM ABQ NLO MW YPZ FGG."} +{"text": "This article has been corrected: The correct author name is given below:Takuro Uchida25075-25088. https://doi.org/10.18632/oncotarget.25308Original article: Oncotarget. 2018; 9:"} +{"text": "There is an error in the Correction published on July 5, 2019. The third author\u2019s name is incorrect in the citation. The publisher apologizes for this error.NORDLAND study. PLoS ONE 14(6): e0218521. https://doi.org/10.1371/journal.pone.0218521.The correct citation is: Catalan-Matamoros D, Lopez-Villegas A, Lappegard KT, Lopez-Liria R (2019) Patients' experiences of remote communication after pacemaker implant: The"} +{"text": "Correction to: BMC Microbiology (2018) 18:191.https://doi.org/10.1186/s12866-018-1329-yFollowing publication of the original article , we haveftp://ftp.ncbi.nih.gov/genomes/genbank/fungi/Stemphylium_lycopersici/latest_assembly_versions/GCA_001191545.1_ASM119154v1/GCA_001191545.1_ASM119154v1_protein.faa.gz) is:The correct citation for the data presented in section \u201cMaterials and Methods - Sequence data and preparation\u201d ."} +{"text": "This article has been corrected: The corrected author name is given below:Ruiqi Chenhttps://doi.org/10.18632/oncotarget.19386Original article: Oncotarget. 2017; 8:84863-84876."} +{"text": "This article has been corrected: The correct Author name is given below:Stephen D. Robinsonhttps://doi.org/10.18632/oncotarget.23378Original article: Oncotarget. 2018; 9:3815-3829."} +{"text": "VOLUME 107107(3) July, page 302http://dx.doi.org/10.5195/jmla.2019.712.Martin ER. Social justice and the medical librarian. J Med Libr Assoc. 2019 Jul;107(3):291\u2013303. DOI: Page 302: The quote by Martin Luther King Jr. should be:\u2014Injustice anywhere is a threat to justice everywhere. Martin Luther King [41]"} +{"text": "This article has been corrected: During production, the author name for this article was given incorrectly. The proper author name is shown below.N. Andres Parrahttps://doi.org/10.18632/oncotarget.26437Original article: Oncotarget. 2018; 9:37125-37136."} +{"text": "Although the Concise Guide represents approximately 400 pages, the material presented is substantially reduced compared to information and links presented on the website. It provides a permanent, citable, point\u2010in\u2010time record that will survive database updates. The full contents of this section can be found at http://onlinelibrary.wiley.com/doi/10.1111/bph.14753. Transporters are one of the six major pharmacological targets into which the Guide is divided, with the others being: G protein\u2010coupled receptors, ion channels, nuclear hormone receptors, catalytic receptors and enzymes. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading. The landscape format of the Concise Guide is designed to facilitate comparison of related targets from material contemporary to mid\u20102019, and supersedes data presented in the 2017/18, 2015/16 and 2013/14 Concise Guides and previous Guides to Receptors and Channels. It is produced in close conjunction with the International Union of Basic and Clinical Pharmacology Committee on Receptor Nomenclature and Drug Classification (NC\u2010IUPHAR), therefore, providing official IUPHAR classification and nomenclature for human drug targets, where appropriate.The Concise Guide to PHARMACOLOGY 2019/20 is the fourth in this series of biennial publications. The Concise Guide provides concise overviews of the key properties of nearly 1800 human drug targets with an emphasis on selective pharmacology (where available), plus links to the open access knowledgebase source of drug targets and their ligands ( Cellular membranes are hydrophobic and, therefore, effective barriers to separate them allowing the formation of gradients, which can be exploited, for example, in the generation of energy. Membrane transporters carry solutes across cell membranes, which would otherwise be impermeable to them. The energy required for active transport processes is obtained from ATP turnover or by exploiting ion gradients.ATP\u2010driven transporters can be divided into three major classes: P\u2010type ATPases; F\u2010type or V\u2010type ATPases and ATP\u2010binding cassette transporters. The first of these, P\u2010type ATPases, are multimeric proteins, which transport (primarily) inorganic cations. The second, F\u2010type or V\u2010type ATPases, are proton\u2010coupled motors, which can function either as transporters or as motors. Last, are ATP\u2010binding cassette transporters, heavily involved in drug disposition as well as transporting endogenous solutes.The second largest family of membrane proteins in the human genome, after the G protein\u2010coupled receptors, are the SLC solute carrier family. Within the solute carrier family, there are a great variety of solutes transported, from simple inorganic ions to amino acids and sugars to relatively complex organic molecules like haem. The solute carrier family includes 65 families of almost 400 members. Many of these overlap in terms of the solutes that they carry. For example, amino acids accumulation is mediated by members of the SLC1, SLC3/7, SLC6, SLC15, SLC16, SLC17, SLC32, SLC36, SLC38 and SLC43 families. Further members of the SLC superfamily regulate ion fluxes at the plasma membrane, or solute transport into and out of cellular organelles. Some SLC family members remain orphan transporters, in as much as a physiological function has yet to be dtermined. Within the SLC super\u2010family, there is an abundance in diversity of structure. Two families (SLC3 and SLC7) only generate functional transporters as heteromeric partners, where one partner is a single TM domain protein. Membrane topology predictions for other families suggest 3,4,6,7,8,9,10,11,12,13 or 14 TM domains. The SLC transporters include members which function as antiports, where solute movement in one direction is balanced by a solute moving in the reverse direction. Symports allow concentration gradients of one solute to allow co\u2010transport of a second solute across a membrane. A third, relatively small group are equilibrative transporters, which allow solutes to travel across membranes down their concentration gradients. A more complex family of transporters, the SLC27 fatty acid transporters also express enzymatic function. Many of the transporters also express electrogenic properties of ion channels.http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=200\u2013 ATP\u2010binding cassette transporters are ubiquitous membrane proteins characterized by active ATP\u2010dependent movement of a range of substrates, including ions, lipids, peptides, steroids. Individual subunits are typically made up of two groups of 6TM\u2010spanning domains, with two nucleotide\u2010binding domains (NBD). The majority of eukaryotic ABC transporters are \u2018full\u2019 transporters incorporating both TM and NBD entities. Some ABCs, notably the ABCD and ABCG families are half\u2010transporters with only a single membrane spanning domain and one NBD, and are only functional as homo\u2010 or heterodimers. Eukaryotic ABC transporters convey substrates from the cytoplasm, either out of the cell or into intracellular organelles. Their role in the efflux of exogenous compounds, notably chemotherapeutic agents, has led to considerable interest.http://www.ncbi.nlm.nih.gov/pubmed/16586097?dopt=AbstractPlus].To date, 12 members of the human ABCA subfamily are identified. They share a high degree of sequence conservation and have been mostly related with lipid trafficking in a wide range of body locations. Mutations in some of these genes have been described to cause severe hereditary diseases related with lipid transport, such as fatal surfactant deficiency or harlequin ichthyosis. In addition, most of them are hypothesized to participate in the subcellular sequestration of drugs, thereby being responsible for the resistance of several carcinoma cell lines against drug treatment .ABCD4 ; https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:70 ; https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:71 and https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:72 (http://www.uniprot.org/uniprot/Q9NUQ8).A further group of ABC transporter\u2010like proteins have been identified to lack membrane spanning regions and are not believed to be functional transporters, but appear to have a role in protein translation [et al. (2015) Peroxisomal ABC transporters: functions and mechanism. Biochem. Soc. Trans. 43: 959\u201065 https://www.ncbi.nlm.nih.gov/pubmed/26517910?dopt=AbstractPlusBaker A Biochem. Soc. Trans. 43: 889\u201093 https://www.ncbi.nlm.nih.gov/pubmed/26517899?dopt=AbstractPlusBeis K. (2015) Structural basis for the mechanism of ABC transporters. et al. (2016) Mammalian drug efflux transporters of the ATP binding cassette (ABC) family in multidrug resistance: A review of the past decade. Cancer Lett. 370: 153\u201064 https://www.ncbi.nlm.nih.gov/pubmed/26499806?dopt=AbstractPlusChen Z et al. (2011) Mammalian peroxisomal ABC transporters: from endogenous substrates to pathology and clinical significance. Br. J. Pharmacol. 164: 1753\u201066 https://www.ncbi.nlm.nih.gov/pubmed/21488864?dopt=AbstractPlusKemp S et al. (2011) The ABCG family of membrane\u2010associated transporters: you don't have to be big to be mighty. Br. J. Pharmacol. 164: 1767\u201079 https://www.ncbi.nlm.nih.gov/pubmed/21175590?dopt=AbstractPlusKerr ID et al. (2017) The Role of Oxysterols in Human Cancer. Trends Endocrinol. Metab. 28: 485\u2010496 https://www.ncbi.nlm.nih.gov/pubmed/28410994?dopt=AbstractPlusKloudova A et al. (2015) Structure and mechanism of ATP\u2010dependent phospholipid transporters. Biochim. Biophys. Acta1850: 461\u2010475 https://www.ncbi.nlm.nih.gov/pubmed/24746984?dopt=AbstractPlusL\u00f3pez\u2010Marqu\u00e9s RL etal. (2016) Impact of Membrane Drug Transporters on Resistance to Small\u2010Molecule Tyrosine Kinase Inhibitors. Trends Pharmacol. Sci. 37: 904\u2010932 https://www.ncbi.nlm.nih.gov/pubmed/27659854?dopt=AbstractPlusNeul C et al. (2017) ABCG2/BCRP: Specific and Nonspecific Modulators. Med Res Rev37: 987\u20101050 https://www.ncbi.nlm.nih.gov/pubmed/28005280?dopt=AbstractPlusPe\u00f1a\u2010Sol\u00f3rzano D et al. (2018) Revisiting the role of ABC transporters in multidrug\u2010resistant cancer. Nat Rev Cancer18: 452\u2010464 [https://www.ncbi.nlm.nih.gov/pubmed/29643473]Robey RW et al. (2017) Targeted pharmacotherapies for defective ABC transporters. Biochem. Pharmacol. 136: 1\u201011 https://www.ncbi.nlm.nih.gov/pubmed/28245962?dopt=AbstractPlusVauthier V 1 or V1) and a membrane complex (Fo or Vo). Within each ATPase complex, the two individual sectors appear to function as connected opposing rotary motors, coupling catalysis of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1713 synthesis or hydrolysis to proton transport. Both the F\u2010type and V\u2010type ATPases have been assigned enzyme commission number http://www.genome.jp/kegg\u2010bin/search_brite?option=\u2010a&search_string=3.6.3.14The F\u2010type (ATP synthase) and the V\u2010type (vacuolar or vesicular proton pump) ATPases, although having distinct sub\u2010cellular locations and roles, exhibit marked similarities in subunit structure and mechanism. They are both composed of a \u2018soluble\u2019 complex (termed F+\u2010transporting), is a mitochondrial membrane\u2010associated multimeric complex consisting of two domains, an F0 channel domain in the membrane and an F1 domain extending into the lumen. Proton transport across the inner mitochondrial membrane is used to drive the synthesis of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1713, although it is also possible for the enzyme to function as an AT\u2010Pase. The ATP5O subunit ), acts as a connector between F1 and F0 motors.The F\u2010type ATPase, also known as ATP synthase or ATP phosphohydrolase .Thehttp://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=157.Information on members of this family may be found in the et al. (2015) Hybrid rotors in F1F(o) ATP synthases: subunit composition, distribution, and physiological significance. Biol. Chem. 396: 1031\u201342 https://www.ncbi.nlm.nih.gov/pubmed/25838297?dopt=AbstractPlusBrandt K Prog. Biophys. Mol. Biol. 119: 94\u2013102 https://www.ncbi.nlm.nih.gov/pubmed/26140992?dopt=AbstractPlusKrah A. (2015) Linking structural features from mitochondrial and bacterial F\u2010type ATP synthases to their distinct mechanisms of ATPase inhibition. et al. (2014) Eukaryotic V\u2010ATPase: novel structural findings and functional insights. Biochim. Biophys. Acta1837: 857\u201379 https://www.ncbi.nlm.nih.gov/pubmed/24508215?dopt=AbstractPlusMarshansky V et al. (2017) Catalytic robustness and torque generation of the F_1\u2010ATPase. Biophys Rev9: 103\u2013118 https://www.ncbi.nlm.nih.gov/pubmed/28424741?dopt=AbstractPlusNoji H et al. (2013) Single\u2010molecule analysis of the rotation of F_1\u2010ATPase under high hydrostatic pressure. Biophys. J. 105: 1635\u201342 https://www.ncbi.nlm.nih.gov/pubmed/24094404?dopt=AbstractPlusOkuno D Phosphorylation\u2010type ATPases (EC 3.6.3.\u2010) are associated with membranes and the transport of ions or phospholipids. Characteristics of the family are the transient phosphorylation of the transporters at an aspartate residue and the interconversion between E1 and E2 conformations in the activity cycle of the transporters, taken to represent \u2018half\u2010channels\u2019 facing the cytoplasm and extracellular/luminal side of the membrane, respectively.Sequence analysis across multiple species allows the definition of five subfamilies, P1\u2010P5. The P1 subfamily includes heavy metal pumps, such as the copper ATPases. The P2 subfamily includes calcium, sodium/potassium and proton/potassium pumps. The P4 and P5 subfamilies include putative phospholipid flippases.+/K+\u2010ATPase is an integral membrane protein which regulates the membrane potential of the cell by maintaining gradients of Na+ and K+ ions across the plasma membrane, also making a small, direct contribution to membrane potential, particularly in cardiac cells. For every molecule of ATP hydrolysed, the Na+/K+\u2010ATPase extrudes three Na+ ions and imports two K+ ions. The active transporter is a heteromultimer with incompletely defined stoichiometry, possibly as tetramers of heterodimers, each consisting of one of four large, ten TM domain catalytic \u03b1 subunits and one of three smaller, single TM domain glycoprotein \u03b2\u2010subunits. Additional protein partners known as FXYD proteins appear to associate with and regulate the activity of the pump.The cell\u2010surface Nahttp://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=158.Information on members of this family may be found in the +/K+\u2010ATPases are inhibited by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4826 and cardiac glycosides, such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4726, as well as potentially endogenous cardiotonic steroids [http://www.ncbi.nlm.nih.gov/pubmed/19325075?dopt=AbstractPlus].Na2+\u2010ATPase (SERCA) is an intracellular membrane\u2010associated pump for sequestering calcium from the cytosol into intracellular organelles, usually associated with the recovery phase following excitation of muscle and nerves.The sarcoplasmic/endoplasmic reticulum Ca2+\u2010ATPase (PMCA) is a cell\u2010surface pump for extruding calcium from the cytosol, usually associated with the recovery phase following excitation of cells. The active pump is a homodimer, each subunit of which is made up of ten TM segments, with cytosolic C\u2010 and N\u2010termini and two large intracellular loops.The plasma membrane Ca2+\u2010ATPases (SPCA) allow accumulation of calcium and manganese in the Golgi apparatus.Secretory pathway Cahttp://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=159.Information on members of this family may be found in the http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4672 has been described to activate SERCA in kidney microsomes [http://www.ncbi.nlm.nih.gov/pubmed/1417961?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5350 [http://www.ncbi.nlm.nih.gov/pubmed/2530215?dopt=AbstractPlus], http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5351 [http://www.ncbi.nlm.nih.gov/pubmed/1832668?dopt=AbstractPlus] and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5486are widely employed to block SERCA. Thapsigargin has also been described to block the TRPV1 vanilloid receptor [http://www.ncbi.nlm.nih.gov/pubmed/12054538?dopt=AbstractPlus].The fungal toxin 2+ while SERCA transports 2 Ca2+.The stoichiometry of flux through the PMCA differs from SERCA, with the PMCA transporting 1 Cahttp://www.ncbi.nlm.nih.gov/pubmed/10615129?dopt=AbstractPlus].Loss\u2010of\u2010function mutations in SPCA1 appear to underlie Hailey\u2010Hailey disease .The Hhttp://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=160.Information on members of this family may be found in the +/K+\u2010ATPase is inhibited by proton pump inhibitors used for treating excessive gastric acid secretion, including http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5487 and a metabolite of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5488.The gastric He.g. https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:798, http://www.uniprot.org/uniprot/O00244).Copper\u2010transporting ATPases convey copper ions across cell\u2010surface and intracellular membranes. They consist of eight TM domains and associate with multiple copper chaperone proteins , https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:30213 (http://www.uniprot.org/uniprot/Q9NQ11), https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:24113 (http://www.uniprot.org/uniprot/Q9H7F0), https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:25422 (http://www.uniprot.org/uniprot/Q4VNC1) and https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:31789 (http://www.uniprot.org/uniprot/Q4VNC0).A further series of structurally\u2010related proteins have been identified in the human genome, with as yet undefined function, including et al. (2016) Na+\u2010K+\u2010ATPase, a new class of plasma membrane receptors. Am. J. Physiol., Cell Physiol. 310: C491\u20135 https://www.ncbi.nlm.nih.gov/pubmed/26791490?dopt=AbstractPlusAperia A et al. (2017) The plasma membrane calcium pumps: focus on the role in (neuro)pathology. Biochem. Biophys. Res. Commun. 483: 1116\u20131124 https://www.ncbi.nlm.nih.gov/pubmed/27480928?dopt=AbstractPlusBrini M Cell Calcium69: 28\u201336 https://www.ncbi.nlm.nih.gov/pubmed/27553475?dopt=AbstractPlusBruce JIE. (2018) Metabolic regulation of the PMCA: Role in cell death and survival. et al. (2017) Cardiac glycosides: From molecular targets to immunogenic cell death. Biochem. Pharmacol. 125: 1\u201311 https://www.ncbi.nlm.nih.gov/pubmed/27553475?dopt=AbstractPlusDiederich M et al. (2016) The calcium\u2010signaling toolkit: Updates needed. Biochim. Biophys. Acta1863: 1337\u201343 https://www.ncbi.nlm.nih.gov/pubmed/26658643?dopt=AbstractPlusDubois C Biochim. Biophys. Acta1853: 2018\u201324 https://www.ncbi.nlm.nih.gov/pubmed/25535949?dopt=AbstractPlusKrebs J. (2015) The plethora of PMCA isoforms: Alternative splicing and differential expression. et al. (2016) Plasma membrane calcium ATPases (PMCAs) as potential targets for the treatment of essential hypertension. Pharmacol. Ther. 159: 23\u201334 https://www.ncbi.nlm.nih.gov/pubmed/26820758?dopt=AbstractPlusLittle R et al. (2015) Structure and mechanism of ATP\u2010dependent phospholipid transporters. Biochim. Biophys. Acta1850: 461\u2013475 https://www.ncbi.nlm.nih.gov/pubmed/24746984?dopt=AbstractPlusL\u00f3pez\u2010Marqu\u00e9s RL IUBMB Life67: 737\u201345 https://www.ncbi.nlm.nih.gov/pubmed/26422816?dopt=AbstractPlusMigocka M. (2015) Copper\u2010transporting ATPases: The evolutionarily conserved machineries for balancing copper in living systems. et al. (2016) Multifaceted plasmamembrane Ca(2+) pumps: From structure to intracellular Ca(2+) handling and cancer. Biochim. Biophys. Acta1863: 1351\u201363 https://www.ncbi.nlm.nih.gov/pubmed/26707182?dopt=AbstractPlusPad\u00e1nyi R et al. (2016) Lipid somersaults: Uncovering the mechanisms of protein\u2010mediated lipid flipping. Prog. Lipid Res. 64: 69\u201384 https://www.ncbi.nlm.nih.gov/pubmed/27528189?dopt=AbstractPlusPomorski TG et al. (2016) P2C\u2010Type ATPases and Their Regulation. Mol. Neurobiol. 53: 1343\u201354 https://www.ncbi.nlm.nih.gov/pubmed/25631710?dopt=AbstractPlusRetamales\u2010Ortega R et al. (2017) Mechanisms of charge transfer in human copper ATPases ATP7A and ATP7B. IUBMB Life69: 218\u2013225 https://www.ncbi.nlm.nih.gov/pubmed/28164426?dopt=AbstractPlusTadini\u2010Buoninsegni F The SLC superfamily of solute carriers is the second largest family of membrane proteins after G protein\u2010coupled receptors, but with a great deal fewer therapeutic drugs that exploit them. As with the ABC transporters, however, they play a major role in drug disposition and so can be hugely influential in determining the clinical efficacy of particular drugs.48 families are identified on the basis of sequence similarities, but many of them overlap in terms of the solutes that they carry. For example, amino acid accumulation is mediated by members of the SLC1, SLC3/7, SLC6, SLC15, SLC16, SLC17, SLC32, SLC36, SLC38 and SLC43. Further members of the SLC superfamily regulate ion fluxes at the plasma membrane, or solute transport into and out of cellular organelles.Within the SLC superfamily, there is an abundance in diversity of structure. Two families (SLC3 and SLC7) only generate functional transporters as heteromeric partners, where one partner is a single TMdomain protein. Membrane topology predictions for other families suggest 3, 4 6, 7, 8, 9, 10, 11, 12, 13, or 14 TM domains.Functionally, members may be divided into those dependent on gradients of ions , exchange of solutes or simple equilibrative gating. For many members, the stoichiometry of transport is not yet established. Furthermore, one family of transporters also possess enzymatic activity (SLC27), while many members function as ion channels (e.g. SLC1A7/EAAT5), which increases the complexity of function of the SLC superfamily.http://www.ncbi.nlm.nih.gov/pubmed/8103691?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/17088867?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/14530974?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/14612154?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/9790568?dopt=AbstractPlus].The SLC1 family of sodium dependent transporters includes the plasma membrane located glutamate transporters and the neutral amino acid transporters ASCT1 and ASCT2 [http://www.ncbi.nlm.nih.gov/pubmed/10734120?dopt=AbstractPlus]. The crystal structure of a glutamate transporter homologue (GltPh) from Pyrococcus horikoshii supports this topology and indicates that the transporter assembles as a trimer, where each monomer is a functional unit capable of substrate permeation reviewed by [http://www.ncbi.nlm.nih.gov/pubmed/20708631?dopt=AbstractPlus]). This structural data is in agreementwith the proposed quaternary structure for EAAT2 [http://www.ncbi.nlm.nih.gov/pubmed/15265858?dopt=AbstractPlus] and several functional studies that propose the monomer is the functional unit . Recent evidence suggests that EAAT3 and EAAT4 may assemble as heterotrimers [http://www.ncbi.nlm.nih.gov/pubmed/21127051?dopt=AbstractPlus]. The activity of glutamate transporters located upon both neurones and glia (predominantly EAAT 1 and 2) serves, dependent upon their location, to regulate excitatory neurotransmission, maintain low ambient extracellular concentrations of glutamate (protecting against excitotoxicity) and provide glutamate for metabolism including the glutamate\u2010glutamine cycle. The http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=138 that maintains the ion gradients that drive transport has been demonstrated to co\u2010assemble with EAAT1 and EAAT2 [http://www.ncbi.nlm.nih.gov/pubmed/19553454?dopt=AbstractPlus]. Recent evidence supports altered glutamate transport and novel roles in brain for splice variants of EAAT1 and EAAT2 . Three patients with dicarboxylic aminoaciduria (DA) were recently found to have loss\u2010of\u2010function mutations in EAAT3 [http://www.ncbi.nlm.nih.gov/pubmed/21123949?dopt=AbstractPlus]. DA is characterized by excessive excretion of the acidic amino acids glutamate and aspartate and EAAT3 is the predominant glutamate/aspartate transporter in the kidney. Enhanced expression of EAAT2 resulting fromadministration of \u00df\u2010lactam antibiotics (e.g. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5326) is neuroprotective and occurs through NF\u2010?B\u2010mediated EAAT2 promoter activation reviewed by [http://www.ncbi.nlm.nih.gov/pubmed/21792905?dopt=AbstractPlus]). http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=86 activation (e.g. by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1056) also leads to enhanced expression of EAAT though promoter activation [http://www.ncbi.nlm.nih.gov/pubmed/17213861?dopt=AbstractPlus]. In addition, several translational activators of EAAT2 have recently been described [http://www.ncbi.nlm.nih.gov/pubmed/20508255?dopt=AbstractPlus] along with treatments that increase the surface expression of EAAT2 , or prevent its down\u2010regulation (e.g. [http://www.ncbi.nlm.nih.gov/pubmed/21730107?dopt=AbstractPlus]). A thermodynamically uncoupled Cl\u2010 flux, activated by Na+ and glutamate (Na+ and aspartate in the case of GltPh [http://www.ncbi.nlm.nih.gov/pubmed/21730107?dopt=AbstractPlus]), is sufficiently large, in the instances of EAAT4 and EAAT5, to influence neuronal excitability . Indeed, it has recently been suggested that the primary function of EAAT5 is as a slow anion channel gated by glutamate, rather than a glutamate transporter [http://www.ncbi.nlm.nih.gov/pubmed/21641307?dopt=AbstractPlus].Glutamate transporters present the unusual structural motif of 8TM segments and 2 re\u2010entrant loops . KB (or Ki) values derived in uptake assays are generally higher (e.g. [http://www.ncbi.nlm.nih.gov/pubmed/9463476?dopt=AbstractPlus]). In addition to acting as a poorly transportable inhibitor of EAAT2, \u20104\u2010methylglutamate, also known as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4317, is a competitive substrate for EAAT1 and additionally is a potent kainate receptor agonist [http://www.ncbi.nlm.nih.gov/pubmed/8996224?dopt=AbstractPlus] which renders the compound unsuitable for autoradiographic localisation of EAATs [http://www.ncbi.nlm.nih.gov/pubmed/17590480?dopt=AbstractPlus]. Similarly, at concentrations that inhibit EAAT2, dihydrokainate binds to kainate receptors [http://www.ncbi.nlm.nih.gov/pubmed/9463476?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5327 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4531 are both non\u2010substrate inhibitors with a preference for EAAT2 over EAAT3 and EAAT1 . http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4626 is a non\u2010substrate inhibitor with modest selectivity for EAAT3 over EAAT1 (>10\u2010fold) and EAAT2 (5\u2010fold) . Analogously, L\u2010\u03b2\u2010threo\u2010benzyl\u2010aspartate (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4625) is a competitive nonsubstrate inhibitor that preferentially blocks EAAT3 versus EAAT1, or EAAT2 [http://www.ncbi.nlm.nih.gov/pubmed/16183084?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4075 demonstrates low affinity binding (KD\u22456.0 \u03bcM) to EAAT1 and EAAT2 in rat brain homogenates [http://www.ncbi.nlm.nih.gov/pubmed/11389172?dopt=AbstractPlus] and EAAT1 in murine astrocyte membranes [http://www.ncbi.nlm.nih.gov/pubmed/14994336?dopt=AbstractPlus], whereas http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4492 binds with high affinity to all EAATs other than EAAT3 [http://www.ncbi.nlm.nih.gov/pubmed/17047096?dopt=AbstractPlus]. The novel isoxazole derivative http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5328 may interact at the same site as TBOA and preferentially inhibit reverse transport of glutamate [http://www.ncbi.nlm.nih.gov/pubmed/18451317?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4573 induces substrate\u2010like currents at EAAT4, but does not elicit heteroexchange of [3H]\u2010aspartate in synaptosome preparations, inconsistentwith the behaviour of a substrate inhibitor [http://www.ncbi.nlm.nih.gov/pubmed/11299317?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5352, a compound isolated from the venom from the spider Parawixia bistriata is a selective enhancer of the glutamate uptake through EAAT2 but not through EAAT1 or EAAT3 . In addition to the agents listed in the table, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4497 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4516 act as non\u2010selective competitive substrate inhibitors of all EAATs. Zn2+ and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2391 are putative endogenous modulators of EAATs with actions that differ across transporter subtypes (reviewed by [http://www.ncbi.nlm.nih.gov/pubmed/15324920?dopt=AbstractPlus]).The K+\u2010dependent exchange of small neutral amino acids such as Ala, Ser, Cys and Thr and their structure is predicted to be similar to that of the glutamate transporters . ASCT1 and ASCT2 also exhibit thermodynamically uncoupled chloride channel activity associated with substrate transport . Whereas EAATs counter\u2010transport K+ (see above) ASCTs do not and their function is independent of the intracellular concentration of K+ [http://www.ncbi.nlm.nih.gov/pubmed/8910405?dopt=AbstractPlus].ASC transporters mediate Nahttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3314 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4704 [http://www.ncbi.nlm.nih.gov/pubmed/14502423?dopt=AbstractPlus]. At low pH (5.5) both ASCT1 and ASCT2 are able to exchange acidic amino acids such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5329 and glutamate . In addition to the inhibitors tabulated above, HgCl2, methylmercury and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5331, at low micromolar concentrations, non\u2010competitively inhibit ASCT2 by covalent modificiation of cysteine residues [http://www.ncbi.nlm.nih.gov/pubmed/20599776?dopt=AbstractPlus].The substrate specificity of ASCT1 may extend to et al. (2007) Transporters for L\u2010glutamate: an update on their molecular pharmacology and pathological involvement. Br. J. Pharmacol. 150: 5\u201317 https://www.ncbi.nlm.nih.gov/pubmed/17088867?dopt=AbstractPlusBeart PM et al. (2016) The importance of the excitatory amino acid transporter 3 (EAAT3). Neurochem. Int. 98: 4\u201318 https://www.ncbi.nlm.nih.gov/pubmed/27233497?dopt=AbstractPlusBjrn\u2010Yoshimoto WE et al. (2016) Molecular physiology of EAAT anion channels. Pflugers Arch. 468: 491\u2013502 https://www.ncbi.nlm.nih.gov/pubmed/26687113?dopt=AbstractPlusFahlke C J. Neurochem. 134: 982\u20131007 https://www.ncbi.nlm.nih.gov/pubmed/26096891?dopt=AbstractPlusFontana AC. (2015) Current approaches to enhance glutamate transporter function and expression. et al. (2014) SLC1 glutamate transporters. Pflugers Arch. 466: 3\u201324 https://www.ncbi.nlm.nih.gov/pubmed/24240778?dopt=AbstractPlusGrewer C et al. (2015) Excitatory amino acid transporters: recent insights into molecular mechanisms, novel modes of modulation and new therapeutic possibilities. Curr Opin Pharmacol20: 116\u201323 https://www.ncbi.nlm.nih.gov/pubmed/25466154?dopt=AbstractPlusJensen AA et al. (2013) The SLC1 high\u2010affinity glutamate and neutral amino acid transporter family. Mol. Aspects Med. 34: 108\u201320 https://www.ncbi.nlm.nih.gov/pubmed/23506861?dopt=AbstractPlusKanai Y et al. (2015) Glutamate transporter EAAT2: regulation, function, and potential as a therapeutic target for neurological and psychiatric disease. Cell. Mol. Life Sci. 72: 3489\u2013506 https://www.ncbi.nlm.nih.gov/pubmed/26033496?dopt=AbstractPlusTakahashi K http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4536, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4654, inositol (e.g. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4495) and related hexoses. Three classes of glucose transporter can be identified, separating GLUT1\u20104 and 14, GLUT6, 8, 10 and 12; and GLUT5, 7, 9 and 11. Modelling suggests a 12 TM membrane topology, with intracellular termini, with functional transporters acting as homodimers or homotetramers.The SLC2 family transports http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4536, but not http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4654, in the direction of the concentration gradient and may be inhibited non\u2010selectively by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4285 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5334. GLUT1 is the major glucose transporter in brain, placenta and erythrocytes, GLUT2 is found in the pancreas, liver and kidneys, GLUT3 is neuronal and placental, while GLUT4 is the insulin\u2010responsive transporter found in skeletal muscle, heart and adipose tissue. GLUT14 appears to result from gene duplication of GLUT3 and is expressed in the testes [http://www.ncbi.nlm.nih.gov/pubmed/12504846?dopt=AbstractPlus].Class I transporters are able to transport http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4654 and appear to be insensitive to http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5334. Class II transporters appear to be predominantly intracellularly located.Class II transporters transport http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4285 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5334 [http://www.ncbi.nlm.nih.gov/pubmed/12135767?dopt=AbstractPlus].Proton\u2010coupled inositol transporters are expressed predominantly in the brain and can be inhibited by IUBMB Life62: 315\u201333 https://www.ncbi.nlm.nih.gov/pubmed/20209635?dopt=AbstractPlusAugustin R. (2010) The protein family of glucose transport facilitators: It's not only about glucose after all. et al. (2014) Signal transduction meets vesicle traffic: the software and hardware of GLUT4 translocation. Am. J. Physiol., Cell Physiol. 306: C879\u201386 https://www.ncbi.nlm.nih.gov/pubmed/24598362?dopt=AbstractPlusKlip A et al. (2009) Themolecular basis of insulin\u2010stimulated glucose uptake: signalling, trafficking and potential drug targets. J. Endocrinol. 203: 1\u201318 https://www.ncbi.nlm.nih.gov/pubmed/19389739?dopt=AbstractPlusLeney SE et al. (2013) The SLC2 (GLUT) family of membrane transporters. Mol. Aspects Med. 34: 121\u201038 https://www.ncbi.nlm.nih.gov/pubmed/23506862?dopt=AbstractPlusMueckler M et al. (2004) The SLC2 family of facilitated hexose and polyol transporters. Pflugers Arch. 447: 480\u20109 https://www.ncbi.nlm.nih.gov/pubmed/12750891?dopt=AbstractPlusUldry M The SLC3 and SLC7 families combine to generate functional transporters, where the subunit composition is a disulphide\u2010linked combination of a heavy chain (SLC3 family) with a light chain (SLC7 family).SLC3 family members are single TM proteins with extensive glycosylation of the exterior C\u2010terminus, which heterodimerize with SLC7 family members in the endoplasmic reticulum and assist in the plasma membrane localization of the transporter.http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=168.Information on members of this family may be found in the \u2010 and sodium\u2010independent transport of cationic amino acids (system y+), apparently as an exchange mechanism. These transporters are sensitive to inhibition by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5335.SLC7 family members may be divided into two major groups: cationic amino acid transporters (CATs) and glycoprotein\u2010associated amino acid transporters (gpaATs). Cationic amino acid transporters are 14 TM proteins, which mediate pHhttp://www.ncbi.nlm.nih.gov/pubmed/12049641?dopt=AbstractPlus], while SLC7A14 has yet to be characterized. Glycoprotein\u2010associated amino acid transporters are 12 TM proteins, which heterodimerize with members of the SLC3 family to act as cell\u2010surface amino acid exchangers.CAT4 appears to be non\u2010functional in heterologous expression Majumdar D et al. (2013) The divergence, actions, roles, and relatives of sodium\u2010coupled bicarbonate transporters. Physiol. Rev. 93: 803\u2010959 [https://www.ncbi.nlm.nih.gov/pubmed/23589833?dopt=AbstractPlus]Parker MD et al. (2016) Band 3, the human red cell chloride/bicarbonate anion exchanger , in a structural context. Biochim. Biophys. Acta1858: 1507\u201032 [https://www.ncbi.nlm.nih.gov/pubmed/27058983?dopt=AbstractPlus]Reithmeier RA et al. (2013) The SLC4 family of bicarbonate (HCO_3\u2010) transporters. Mol. Aspects Med. 34: 159\u201082 [https://www.ncbi.nlm.nih.gov/pubmed/23506864?dopt=AbstractPlus]Romero MF et al. (2015) Regulators of Slc4 bicarbonate transporter activity. Front Physiol6: 166 [https://www.ncbi.nlm.nih.gov/pubmed/26124722?dopt=AbstractPlus]Thornell IM +/substrate co\u2010transporters for glucose (e.g. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4551), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4536, monocarboxylates, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4495 and I\u2010 . Members of the SLC5 and SLC6 families, along with other unrelated Na+ cotransporters (i.e. Mhp1 and BetP), share a common structural core that contains an inverted repeat of 5TM \u03b1\u2010helical domains [http://www.ncbi.nlm.nih.gov/pubmed/19631523?dopt=AbstractPlus].The SLC5 family of sodium\u2010dependent glucose transporters includes, in mammals, the Nahttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4757, a natural dihydrocholine glucoside, that exhibits modest selectivity towards SGLT2 (see [http://www.ncbi.nlm.nih.gov/pubmed/21527736?dopt=AbstractPlus] for an extensive review). SGLT1 is predominantly expressed in the small intestine, mediating the absorption of glucose (e.g. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4536), but also occurs in the brain, heart and in the late proximal straight tubule of the kidney. The expression of SGLT2 is almost exclusively restricted to the early proximal convoluted tubule of the kidney, where it is largely responsible for the renal reabsorption of glucose. SGLT3 is not a transporter but instead acts as a glucosensor generating an inwardly directed flux of Na+ that causes membrane depolarization [http://www.ncbi.nlm.nih.gov/pubmed/13130073?dopt=AbstractPlus].Detailed characterisation of members of the hexose transporter family is limited to SGLT1, 2 and 3, which are all inhibited in a competitive manner by http://www.ncbi.nlm.nih.gov/pubmed/21527736?dopt=AbstractPlus] for a detailed quantification). Although SGLT1 and SGLT2 have been described as high\u2010 and lowaffinity sodium glucose co\u2010transporters, respectively, recent work suggests that they have a similar affinity for glucose under physiological conditions [http://www.ncbi.nlm.nih.gov/pubmed/20980548?dopt=AbstractPlus]. Selective blockers of SGLT2, and thus blocking 50% of renal glucose reabsorption, are in development for the treatment of diabetes (e.g. [http://www.ncbi.nlm.nih.gov/pubmed/20508640?dopt=AbstractPlus]).Recognition and transport of substrate by SGLTs requires that the sugar is a pyranose. De\u2010oxyglucose derivatives have reduced affinity for SGLT1, but the replacement of the sugar equatorial hydroxyl group by fluorine at some positions, excepting C2 and C3, is tolerated proteins, (which are members of the SLC44 family) with weak Na+ dependence have been described [http://www.ncbi.nlm.nih.gov/pubmed/15715662?dopt=AbstractPlus].The high affinity, hemicholinium\u20103\u2010sensitive, choline transporter (CHT) is expressed mainly in cholinergic neurones on nerve cell terminals and synaptic vesicles . In autonomic neurones, expression of CHT requires an activity\u2010dependent retrograde signal from postsynaptic neurones [i and KD values for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4494 listed in the table are for human CHT expressed in Xenopus laevis oocytes [http://www.ncbi.nlm.nih.gov/pubmed/11068039?dopt=AbstractPlus], or COS\u20107 cells [http://www.ncbi.nlm.nih.gov/pubmed/11027560?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5502 is a substrate for CHT that causes covalent modification and irreversible inactivation of the transporter. Several exogenous substances (e.g. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4760) that are substrates for CHT act as precursors to cholinergic false transmitters.K\u2010 within thyrocytes. Transport of I\u2010 by NIS from the blood across the basolateral membrane followed by apical efflux into the colloidal lumen, mediated at least in part by pendrin (SLC22A4), and most likely not SMCT1 (SLC5A8) as once thought, provides the I\u2010 required for the synthesis of the thyroid hormones triiodothyronine (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2634) and thyroxine (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2635) [http://www.ncbi.nlm.nih.gov/pubmed/19196800?dopt=AbstractPlus]. NIS is also expressed in the salivary glands, gastric mucosa, intestinal enterocytes and lactating breast. NIS mediates I\u2010 absorption in the intestine and I\u2010 secretion into the milk. SMVT is expressed on the apical membrane of intestinal enterocytes and colonocytes and is the main system responsible for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4787 (vitamin H) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4668 (vitamin B5) uptake in humans [http://www.ncbi.nlm.nih.gov/pubmed/19056639?dopt=AbstractPlus]. SMVT located in kidney proximal tubule epithelial cells mediates the reabsorption of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4787 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4668. SMCT1 (SLC5A8), which transports a wide range of monocarboxylates, is expressed in the apical membrane of epithelia of the small intestine, colon, kidney, brain neurones and the retinal pigment epithelium [http://www.ncbi.nlm.nih.gov/pubmed/18446519?dopt=AbstractPlus]. SMCT2 (SLC5A12) also localises to the apical membrane of kidney, intestine, and colon, but in the brain and retina is restricted to astrocytes and M\u00fcller cells, respectively [http://www.ncbi.nlm.nih.gov/pubmed/18446519?dopt=AbstractPlus]. SMCT1 is a high\u2010affinity transporter whereas SMCT2 is a lowaffinity transporter. The physiological substrates for SMCT1 and SMCT2 are lactate (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2932 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2934), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4809, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1062, and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1588 in non\u2010colonic tissues such as the kidney. SMCT1 is also likely to be the principal transporter for the absorption of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1588 (vitamin B3) in the intestine and kidney [http://www.ncbi.nlm.nih.gov/pubmed/15651982?dopt=AbstractPlus]. In the small intestine and colon, the physiological substrates for these transporters are http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1588 and the shortchain fatty acids http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1058, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1062, and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1059 that are produced by bacterial fermentation of dietary fiber [http://www.ncbi.nlm.nih.gov/pubmed/14966140?dopt=AbstractPlus]. In the kidney, SMCT2 is responsible for the bulk absorption of lactate because of its low\u2010affinity/high\u2010capacity nature. Absence of both transporters in the kidney leads to massive excretion of lactate in urine and consequently drastic decrease in the circulating levels of lactate in blood [http://www.ncbi.nlm.nih.gov/pubmed/16873376?dopt=AbstractPlus]. SMCT1 also functions as a tumour suppressor in the colon as well as in various other non\u2010colonic tissues [http://www.ncbi.nlm.nih.gov/pubmed/18992769?dopt=AbstractPlus]. The tumour\u2010suppressive function of SMCT1 is based on its ability to transport http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4809, an inhibitor of histone deacetylases, into cells in non\u2010colonic tissues [http://www.ncbi.nlm.nih.gov/pubmed/17178845?dopt=AbstractPlus]; in the colon, the ability of SMCT1 to transport http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1059 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1062, also inhibitors of histone deacetylases, underlies the tumour\u2010suppressive function of this transporter . The ability of SMCT1 to promote histone acetylase inhibition through accumulation of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1059 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1062 in immune cells is also responsible for suppression of dendritic cell development in the colon [http://www.ncbi.nlm.nih.gov/pubmed/20601425?dopt=AbstractPlus].The sodium\u2010iodide symporter (NIS) is an iodide transporter found principally in the thyroid gland where it mediates the accumulation of I\u2010, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4524, thiocyanate and NO3\u2010 are competitive substrate inhibitors of NIS [http://www.ncbi.nlm.nih.gov/pubmed/18077370?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4822 appears to act as a competitive substrate inhibitor of SMVT [http://www.ncbi.nlm.nih.gov/pubmed/10329687?dopt=AbstractPlus] and the anticonvulsant drugs http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5338 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5339 competitively block the transport of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4787 by brush border vesicles prepared from human intestine [http://www.ncbi.nlm.nih.gov/pubmed/2911998?dopt=AbstractPlus].I+\u2010coupled SMIT1 and SMIT2 tabulated below and the third is proton\u2010coupled HMIT (SLC2A13). SMIT1 and SMIT2 have a widespread and overlapping tissue location but in polarized cells, such as the Madin\u2010 Darby canine kidney cell line, they segregate to the basolateral and apical membranes, respectively [http://www.ncbi.nlm.nih.gov/pubmed/15181167?dopt=AbstractPlus]. In the nephron, SMIT1 mediates http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4495 uptake as a \u2018compatible osmolyte\u2019 when inner medullary tubules are exposed to increases in extracellular osmolality, whilst SMIT2 mediates the reabsorption of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4495 from the filtrate. In some species apically located SMIT2 is responsible for the uptake of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4495 from the intestinal lumen [http://www.ncbi.nlm.nih.gov/pubmed/17932225?dopt=AbstractPlus].Three different mammalian myo\u2010inositol cotransporters are currently known; two are the Nahttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4645, but SMIT1 does not. In addition, whereas SMIT1 transports both http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4724 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4720 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4722 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4721, SMIT2 transports only the D\u2010isomers of these sugars . Thus the substrate specificities of SMIT1 (for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4721) and SMIT2 (for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4645) allow discrimination between the two SMITs. Human SMIT2 appears not to transport glucose [http://www.ncbi.nlm.nih.gov/pubmed/19032932?dopt=AbstractPlus].The data tabulated are those for dog SMIT1 and rabbit SMIT2. SMIT2 transports et al. (2017) Renal, metabolic and cardiovascular considerations of SGLT2 inhibition. Nat Rev Nephrol13: 11\u201026 https://www.ncbi.nlm.nih.gov/pubmed/27941935?dopt=AbstractPlusDeFronzo RA +\u2010D\u2010glucose cotransporters SGLT1 and SGLT2 are targets for the treatment of diabetes and cancer. Pharmacol. Ther. 170: 148\u2010165 https://www.ncbi.nlm.nih.gov/pubmed/27773781?dopt=AbstractPlusKoepsell H. (2017) The Naet al. (2016) Intestinal SGLT1 inmetabolic health and disease. Am. J. Physiol. Gastrointest. Liver Physiol. 310: G887\u201098 https://www.ncbi.nlm.nih.gov/pubmed/27012770?dopt=AbstractPlusLehmann A Mol. Aspects Med. 34: 183\u201096 https://www.ncbi.nlm.nih.gov/pubmed/23506865?dopt=AbstractPlusWright EM. (2013) Glucose transport families SLC5 and SLC50. et al. (2011) Biology of human sodium glucose transporters. Physiol. Rev. 91: 733\u201094 https://www.ncbi.nlm.nih.gov/pubmed/21527736?dopt=AbstractPlusWright EM http://www.ncbi.nlm.nih.gov/pubmed/16540203?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/12719981?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/21752877?dopt=AbstractPlus] are primarily plasma membrane located and may be divided into four subfamilies that transport monoamines, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1067, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=727 and neutral amino acids, plus the related bacterial NSS transporters [http://www.ncbi.nlm.nih.gov/pubmed/19022853?dopt=AbstractPlus]. The members of this superfamily share a structural motif of 10 TM segments that has been observed in crystal structures of the NSS bacterial homolog LeuTAa, a Na+\u2010dependent amino acid transporter from Aquiflex aeolicus [http://www.ncbi.nlm.nih.gov/pubmed/16041361?dopt=AbstractPlus] and in several other transporter families structurally related to LeuT [http://www.ncbi.nlm.nih.gov/pubmed/19996368?dopt=AbstractPlus].Members of the solute carrier family 6 (SLC6) of sodium\u2010 and (sometimes chloride\u2010) dependent neurotransmitter transporters .http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1067 are sufficient to sustain tonic inhibition mediated by high affinity GABAA receptors in certain neuronal populations [http://www.ncbi.nlm.nih.gov/pubmed/15111008?dopt=AbstractPlus]. GAT1 is the predominant GABA transporter in the brain and occurs primarily upon the terminals of presynaptic neurones and to a much lesser extent upon distal astocytic processes that are in proximity to axons terminals. GAT3 resides predominantly on distal astrocytic terminals that are close to the GABAergic synapse. By contrast, BGT1 occupies an extrasynaptic location possibly along with GAT2 which has limited expression in the brain [http://www.ncbi.nlm.nih.gov/pubmed/20026354?dopt=AbstractPlus]. TauT is a high affinity taurine transporter involved in osmotic balance that occurs in the brain and non\u2010neuronal tissues, such as the kidney, brush border membrane of the intestine and blood brain barrier . CT1, which transports http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4496, has a ubiquitous expression pattern, often co\u2010localizing with creatine kinase [http://www.ncbi.nlm.nih.gov/pubmed/12719981?dopt=AbstractPlus].The activity of GABA\u2010transporters located predominantly upon neurones (GAT\u20101), glia (GAT\u20103) or both serves to terminate phasic GABA\u2010ergic transmission, maintain low ambient extracellular concentrations of GABA, and recycle GABA for reuse by neurones. Nonetheless, ambient concentrations of 50 values for the human orthologue [http://www.ncbi.nlm.nih.gov/pubmed/19275529?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4677 is only weakly selective for GAT 2 and GAT3, with IC50 values in the range 22 to >30 \u03bcM at GAT1 and BGT1, whereas http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4610 has at least an order of magnitude selectivity for BGT1 for reviews]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5489 is a recently described compound that displays 20\u2010fold selectivity for GAT3 over GAT1 [http://www.ncbi.nlm.nih.gov/pubmed/16766089?dopt=AbstractPlus]. In addition to the inhibitors listed, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5490 is a moderately potent, though non\u2010selective, inhibitor of all cloned GABA transporters . Diaryloxime and diarylvinyl ether derivatives of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4564 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4691 that potently inhibit the uptake of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5410 into rat synaptosomes have been described [http://www.ncbi.nlm.nih.gov/pubmed/10479278?dopt=AbstractPlus]. Several derivatives of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5418 (e.g.http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5419 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5420) demonstrate selectivity as blockers of astroglial, versus neuronal, uptake of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1067 for reviews]. GAT3 is inhibited by physiologically relevant concentrations of Zn2+ [http://www.ncbi.nlm.nih.gov/pubmed/15829583?dopt=AbstractPlus]. Taut transports http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1067, but with low affinity, but CT1 does not, although it can be engineered to do so by mutagenesis guided by LeuT as a structural template [http://www.ncbi.nlm.nih.gov/pubmed/17400549?dopt=AbstractPlus]. Although inhibitors of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4496 transport by CT1 are known (e.g. [http://www.ncbi.nlm.nih.gov/pubmed/9882430?dopt=AbstractPlus]) they insufficiently characterized to be included in the table.The IC50 values for GAT1\u20104 reported in the table reflect the range reported in the literature from studies of both human and mouse transporters. There is a tendency towards lower IChttp://www.ncbi.nlm.nih.gov/pubmed/16417482?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/15950877?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/16722246?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/12354619?dopt=AbstractPlus] for reviews). GlyT1 transporter isoforms expressed in glia surrounding glutamatergic synapses regulate synaptic http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=727 concentrations influencing NMDA receptor\u2010mediated neurotransmission , but also are important, in early neonatal life, for regulating glycine concentrations at inhibitory glycinergic synapses [http://www.ncbi.nlm.nih.gov/pubmed/14622582?dopt=AbstractPlus]. Homozygous mice engineered to totally lack GlyT1 exhibit severe respiratory and motor deficiencies due to hyperactive glycinergic signalling and die within the first postnatal day . Disruption of GlyT1 restricted to forebrain neurones is associated with enhancement of EPSCs mediated by NMDA receptors and behaviours that are suggestive of a promnesic action [http://www.ncbi.nlm.nih.gov/pubmed/16554468?dopt=AbstractPlus]. GlyT2 transporters localised on the axons and boutons of glycinergic neurones appear crucial for efficient transmitter loading of synaptic vesicles but may not be essential for the termination of inhibitory neurotransmission . Mice in which GlyT2 has been deleted develop a fatal hyperekplexia phenotype during the second postnatal week [http://www.ncbi.nlm.nih.gov/pubmed/14622583?dopt=AbstractPlus] and mutations in the human gene encoding GlyT2 (SLC6A5) have been identified in patients with hyperekplexia (reviewed by [http://www.ncbi.nlm.nih.gov/pubmed/18707791?dopt=AbstractPlus]). ATB0+ (SLC6A14) is a transporter for numerous dipolar and cationic amino acids and thus has a much broader substrate specificity than the glycine transporters alongside which it is grouped on the basis of structural similarity [http://www.ncbi.nlm.nih.gov/pubmed/12719981?dopt=AbstractPlus]. ATB0+ is expressed in various peripheral tissues [http://www.ncbi.nlm.nih.gov/pubmed/12719981?dopt=AbstractPlus]. By contrast PROT (SLC6A7), which is expressed only in brain in association with a subset of excitatory nerve terminals, shows specificity for the transport of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3314.Two gene products, GlyT1 and GlyT2, are known that give rise to transporters that are predominantly located on glia and neurones, respectively. Five variants of GlyT1 differing in their N\u2010 and C\u2010termini are generated by alternative promoter usage and splicing, and three splice variants of GlyT2 have also been identified [http://www.ncbi.nlm.nih.gov/pubmed/17383967?dopt=AbstractPlus]. Inhibition of GLYT1 by the sarcosine derivatives http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4620, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4601 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4600 is non\u2010competitive . IC50 values for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4600 reported in the literature vary, most likely due to differences in assay conditions . The tricyclic antidepressant http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=201 weakly inhibits GlyT2 (IC50 92 \u03bcM) with approximately 10\u2010fold selectivity over GlyT1 [http://www.ncbi.nlm.nih.gov/pubmed/10694221?dopt=AbstractPlus]. The endogenous lipids http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2391 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2364 exert opposing effects upon GlyT1a, inhibiting (IC50 2 \u03bcM) and potentiating (EC50 13 \u03bcM) transport currents, respectively [http://www.ncbi.nlm.nih.gov/pubmed/12558979?dopt=AbstractPlus]. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5493, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5494 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5495 have been described as endogenous non\u2010competitive inhibitors of GlyT2a, but not GlyT1b . Protons [http://www.ncbi.nlm.nih.gov/pubmed/10860934?dopt=AbstractPlus] and Zn2+ [http://www.ncbi.nlm.nih.gov/pubmed/15031290?dopt=AbstractPlus] act as non\u2010competitive inhibitors of GlyT1b, with IC50 values of 100 nM and 10 \u03bcM respectively, but neither ion affects GlyT2 (reviewed by [http://www.ncbi.nlm.nih.gov/pubmed/15324920?dopt=AbstractPlus]). Glycine transport by GLYT1 is inhibited by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5212, whereas GLYT2 transport is stimulated (both in the presence of Na+) [http://www.ncbi.nlm.nih.gov/pubmed/21574997?dopt=AbstractPlus].0AT1 (SLC6A19), SLC6A18, SLC6A20). Others may function as transporters for neurotransmitters or their precursors [http://www.ncbi.nlm.nih.gov/pubmed/18400692?dopt=AbstractPlus]. B0AT1 has been proposed as a drug target to treat phenylketonuria [http://www.ncbi.nlm.nih.gov/pubmed/30046012?dopt=AbstractPlus].Certain members of neutral amino acid transport family are expressed upon the apical surface of epithelial cells and are important for the absorption of amino acids from the duodenum, jejunum and ileum and their reabsorption within the proximal tubule of the nephron Kinase\u2010dependent Regulation of Monoamine Neurotransmitter Transporters. Pharmacol. Rev. 68: 888\u2010953 [https://www.ncbi.nlm.nih.gov/pubmed/27591044?dopt=AbstractPlus]Bermingham DP et al. (2012) The solute carrier 6 family of transporters. Br. J. Pharmacol. 167: 256\u201078 [https://www.ncbi.nlm.nih.gov/pubmed/22519513?dopt=AbstractPlus]Br\u00f6er S et al. (2015) Creatine biosynthesis and transport in health and disease. Biochimie119: 146\u201065 [https://www.ncbi.nlm.nih.gov/pubmed/26542286?dopt=AbstractPlus]Joncquel\u2010Chevalier Curt M et al. (2017) Membrane transporters as mediators of synaptic dopamine dynamics: implications for disease. Eur. J. Neurosci. 45: 20\u201033 [https://www.ncbi.nlm.nih.gov/pubmed/27520881?dopt=AbstractPlus]Lohr KM 2+\u2010ATPase (http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=138#159_overview) and sarcoplasmic/endoplasmic reticulum Ca2+\u2010ATPase (http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=138), as well as the sodium/potassium/calcium exchangers , NCX allow recovery of intracellular calcium back to basal levels after cellular stimulation. When intracellular sodium ion levels rise, for example, following depolarisation, these transporters can operate in the reverse direction to allow calcium influx and sodium efflux, as an electrogenic mechanism. Structural modelling suggests the presence of 9 TM segments, with a large intracellular loop between the fifth and sixth TM segments.The sodium/calcium exchangers (NCX) use the extracellular sodium concentration to facilitate the extrusion of calcium out of the cell. Alongside the plasma membrane Cahttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4597 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4593 act as non\u2010selective NCX inhibitors, while http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4617, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4232, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4666, and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6481 [http://www.ncbi.nlm.nih.gov/pubmed/23647096?dopt=AbstractPlus] act to inhibit NCX function with varying degrees of selectivity. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=8438 is a selective NCX3 inhibitor [http://www.ncbi.nlm.nih.gov/pubmed/25942323?dopt=AbstractPlus] and and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=9484 inhibits NCX3 preferentially over other isoforms .Although subtype\u2010selective inhibitors of NCX function are not widely available, et al. (2016) Structure\u2010Functional Basis of Ion Transport in Sodium\u2010Calcium Exchanger (NCX) Proteins. Int J Mol Sci17: [https://www.ncbi.nlm.nih.gov/pubmed/27879668?dopt=AbstractPlus]Giladi M Mol. Aspects Med. 34: 220\u201035 [https://www.ncbi.nlm.nih.gov/pubmed/23506867?dopt=AbstractPlus]Khananshvili D. (2013) The SLC8 gene family of sodium\u2010calcium exchangers (NCX) \u2010 structure, function, and regulation in health and disease. Biochem. Biophys. Res. Commun. 460: 50\u20102 [https://www.ncbi.nlm.nih.gov/pubmed/25998733?dopt=AbstractPlus]Sekler I. (2015) Standing of giants shoulders the story of the mitochondrial Na(+)Ca(2+) exchanger. + (in) : 1 H+ (out). Several isoforms, NHE6, NHE7, NHE8 and NHE9 appear to locate on intracellularmembranes . Li+ and NH4+, but not K+, ions may also be transported by some isoforms. Modelling of the topology of these transporters indicates 12 TM regions with an extended intracellular C\u2010terminus containing multiple regulatory sites. NHE1 is considered to be a ubiquitously\u2010expressed \u2018housekeeping\u2019 transporter. NHE3 is highly expressed in the intestine and kidneys and regulate sodium movements in those tissues. NHE10 is present in sperm [http://www.ncbi.nlm.nih.gov/pubmed/14634667?dopt=AbstractPlus] and osteoclasts [http://www.ncbi.nlm.nih.gov/pubmed/18269914?dopt=AbstractPlus]; gene disruption results in infertile male mice [http://www.ncbi.nlm.nih.gov/pubmed/14634667?dopt=AbstractPlus].Sodium/hydrogen exchangers or sodium/proton antiports are a family of transporters that maintain cellular pH by utilising the sodium gradient across the plasma membrane to extrude protons produced by metabolism, in a stoichiometry of 1 Nahttp://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=181.Information on members of this family may be found in the + binding site. The more selective amiloride analogues http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4595 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4186 exhibit a rank order of affinity of inhibition of NHE1 > NHE2 > NHE3 .Analogues of the non\u2010selective cation transport inhibitor amiloride appear to inhibit NHE function through competitive inhibition of the extracellular Naet al. (2013) SLC9/NHE gene family, a plasma membrane and organellar family of Na+/H+ exchangers. Mol. Aspects Med. 34: 236\u201051 [https://www.ncbi.nlm.nih.gov/pubmed/23506868?dopt=AbstractPlus]Donowitz M et al. (2011) Regulation of electroneutral NaCl absorption by the small intestine. Annu. Rev. Physiol. 73: 261\u201081 [https://www.ncbi.nlm.nih.gov/pubmed/21054167?dopt=AbstractPlus]Kato A et al. (2011) Organellar Na+/H+ exchangers: novel players in organelle pH regulation and their emerging functions. Biochemistry50: 443\u201050 [https://www.ncbi.nlm.nih.gov/pubmed/21171650?dopt=AbstractPlus]Ohgaki R et al. (2015) Na+\u2010H+ exchanger\u20101 (NHE1) regulation in kidney proximal tubule. Cell. Mol. Life Sci. 72: 2061\u201074 [https://www.ncbi.nlm.nih.gov/pubmed/25680790?dopt=AbstractPlus]Parker MD et al. (2014) Intracellular pH regulation by acid\u2010base transporters in mammalian neurons. Front Physiol5: 43 [https://www.ncbi.nlm.nih.gov/pubmed/24592239?dopt=AbstractPlus]Ruffin VA http://www.ncbi.nlm.nih.gov/pubmed/19498215?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/20103563?dopt=AbstractPlus]. SLC10A6 (SOAT) functions as a sodium\u2010dependent transporter of sulphated solutes including sulfphated steroids and bile acids . Transport function has not yet been demonstrated for the 4 remaining members of the SLC10 family, SLC10A3 (P3), SLC10A4 (P4), SLC10A5 (P5), and SLC10A7 (P7), and the identity of their endogenous substrates remain unknown . Members of the SLC10 family are predicted to have seven transmembrane domains with an extracellular N\u2010terminus and cytoplasmic C\u2010terminus .The SLC10 family transport bile acids, sulphated solutes, and other xenobiotics in a sodium\u2010dependent manner. The founding members, SLC10A1 (NTCP) and SLC10A2 (ASBT) function, along with members of the ABC transporter family and the organic solute transporter obligate heterodimer OSTa:OST\u00df (SLC51), to maintain the enterohepatic circulation of bile acids [http://www.ncbi.nlm.nih.gov/pubmed/18355966?dopt=AbstractPlus] or SLC10A7 [http://www.ncbi.nlm.nih.gov/pubmed/17628207?dopt=AbstractPlus] failed to exhibit significant transport of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4547, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4290, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4528 or http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4551. SLC10A4 has recently been suggested to associate with neuronal vesicles [http://www.ncbi.nlm.nih.gov/pubmed/21742018?dopt=AbstractPlus].Heterologously expressed SLC10A4 [et al. (2014) Sodium\u2010dependent bile salt transporters of the SLC10A transporter family: more than solute transporters. Pflugers Arch. 466: 77\u201089 [https://www.ncbi.nlm.nih.gov/pubmed/24196564?dopt=AbstractPlus]Anwer MS et al. (2013) The solute carrier family 10 (SLC10): beyond bile acid transport. Mol. Aspects Med. 34: 252\u201069 [https://www.ncbi.nlm.nih.gov/pubmed/23506869?dopt=AbstractPlus]Claro da Silva T Dig Dis 35: 261\u2010266 [https://www.ncbi.nlm.nih.gov/pubmed/28249269?dopt=AbstractPlus]Dawson PA. (2017) Roles of Ileal ASBT and OSTa\u2010OST\u00df in Regulating Bile Acid Signaling. et al. (2013) Transport and biological activities of bile acids. Int. J. Biochem. Cell Biol. 45: 1389\u201098 [https://www.ncbi.nlm.nih.gov/pubmed/23603607?dopt=AbstractPlus]Zwicker BL The family of proton\u2010coupled metal ion transporters are responsible for movements of divalent cations, particularly ferrous and manganese ions, across the cell membrane (SLC11A2/DMT1) and across endosomal (SLC11A2/DMT1) or lysosomal/phagosomal membranes (SLC11A1/NRAMP1), dependent on proton transport. Both proteins appear to have 12 TM regions and cytoplasmic N\u2010 and C\u2010 termini. NRAMP1 is involved in antimicrobial action in macrophages, although its precise mechanism is undefined. Facilitated diffusion of divalent cations into phagosomes may increase intravesicular free radicals to damage the pathogen. Alternatively, export of divalent cations from the phagosome may deprive the pathogen of essential enzyme cofactors. SLC11A2/DMT1 is more widely expressed and appears to assist in divalent cation assimilation from the diet, as well as in phagocytotic cells.http://omim.org/entry/607948). Loss\u2010of\u2010function mutations in DMT1 are associated with microcytic anemia (http://omim.org/entry/206100).Loss\u2010of\u2010function mutations in NRAMP1 are associated with increased susceptibility to microbial infection Iron entry in neurons and astrocytes: a link with synaptic activity. Front Mol Neurosci8: 18 [https://www.ncbi.nlm.nih.gov/pubmed/26089776?dopt=AbstractPlus]Codazzi F et al. (2013) Mammalian iron transporters: families SLC11 and SLC40. Mol. Aspects Med. 34: 270\u201087 [https://www.ncbi.nlm.nih.gov/pubmed/23506870?dopt=AbstractPlus]Montalbetti N J. Biol. Chem. 290: 18984\u201090 [https://www.ncbi.nlm.nih.gov/pubmed/26055722?dopt=AbstractPlus]Wessling\u2010Resnick M. (2015) Nramp1 and Other Transporters Involved in Metal Withholding during Infection. et al. (2012) Regulation of brain iron and copper homeostasis by brain barrier systems: implication in neurodegenerative diseases. Pharmacol. Ther. 133: 177\u201088 [https://www.ncbi.nlm.nih.gov/pubmed/22115751?dopt=AbstractPlus]Zheng W The SLC12 family of chloride transporters contribute to ion fluxes across a variety of tissues, particularly in the kidney and choroid plexus of the brain. Within this family, further subfamilies are identifiable: NKCC1, NKCC2 and NCC constitute a group of therapeutically\u2010relevant transporters, targets for loop and thiazide diuretics. These 12 TM proteins exhibit cytoplasmic termini and an extended extracellular loop at TM7/8 and are kidneyspecific (NKCC2 and NCC) or show a more widespread distribution (NKCC1). A second family, the K\u2010Cl co\u2010transporters are also 12 TM domain proteins with cytoplasmic termini, but with an extended extracellular loop at TM 5/6. CCC6 exhibits structural similarities with the K\u2010Cl co\u2010transporters, while CCC9 is divergent, with 11 TM domains and a cytoplasmic N\u2010terminus and extracellular C\u2010terminus.http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4589 is able to differentiate KCC isoforms from NKCC and NCC transporters, but also inhibits CFTR [http://www.ncbi.nlm.nih.gov/pubmed/11527541?dopt=AbstractPlus].et al. (2013) The SLC12 family of electroneutral cation\u2010coupled chloride cotransporters. Mol. Aspects Med. 34: 288\u201098 [https://www.ncbi.nlm.nih.gov/pubmed/23506871?dopt=AbstractPlus]Arroyo JP et al. (2017) Regulation of renal Na\u2010(K)\u2010Cl cotransporters by vasopressin. Pflugers Arch. 469: 889\u2010897 [https://www.ncbi.nlm.nih.gov/pubmed/28577072?dopt=AbstractPlus]Bachmann S et al. (2016) Physiological role of SLC12 family members in the kidney. Am. J. Physiol. Renal Physiol. 311: F131\u201044 [https://www.ncbi.nlm.nih.gov/pubmed/27097893?dopt=AbstractPlus]Baz\u00faa\u2010Valenti S et al. (2016) Everything we always wanted to know about furosemide but were afraid to ask. Am. J. Physiol. Renal Physiol. 310: F958\u201071 [https://www.ncbi.nlm.nih.gov/pubmed/26911852?dopt=AbstractPlus]Huang X et al. (2015) K\u2010Cl cotransporters, cell volume homeostasis, and neurological disease. Trends Mol Med21: 513\u201023 [https://www.ncbi.nlm.nih.gov/pubmed/26142773?dopt=AbstractPlus]Kahle KT et al. (2017) Chloride co\u2010transporters as possible therapeutic targets for stroke. J. Neurochem. 140: 195\u2010209 [https://www.ncbi.nlm.nih.gov/pubmed/27861901?dopt=AbstractPlus]Mart\u00edn\u2010Arag\u00f3n Baudel MA Within the SLC13 family, two groups of transporters may be differentiated on the basis of the substrates transported: NaS1 and NaS2 convey sulphate, while NaC1\u20103 transport carboxylates. NaS1 and NaS2 transporters are made up of 13 TM domains, with an intracellular N terminus and are electrogenic with physiological roles in the intestine, kidney and placenta. NaC1, NaC2 and NaC3 are made up of 11 TM domains with an intracellular N terminus and are electrogenic, with physiological roles in the kidney and liver.et al. (2013) SLC13 family of Na+\u2010coupled di\u2010 and tri\u2010carboxylate/sulfate transporters. Mol. Aspects Med. 34: 299\u2010312 [https://www.ncbi.nlm.nih.gov/pubmed/23506872?dopt=AbstractPlus]Bergeron MJ +\u2010sulfate cotransporter SLC13A1. Pflugers Arch. 466: 131\u20107 [https://www.ncbi.nlm.nih.gov/pubmed/24193406?dopt=AbstractPlus]Markovich D. (2014) NaPflugers Arch. 466: 119\u201030 [https://www.ncbi.nlm.nih.gov/pubmed/24114175?dopt=AbstractPlus]Pajor AM. (2014) Sodium\u2010coupled dicarboxylate and citrate transporters from the SLC13 family. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4539 movement down its concentration gradient. Multiple splice variants of these transporters have been identified; for UT\u2010A transporters, in particular, there is evidence for cell\u2010specific expression of these variants with functional impact [http://www.ncbi.nlm.nih.gov/pubmed/21449978?dopt=AbstractPlus]. Topographical modelling suggests that the majority of the variants of SLC14 transporters have 10 TM domains, with a glycosylated extracellular loop at TM5/6, and intracellular C\u2010 and N\u2010termini. The UT\u2010A1 splice variant, exceptionally, has 20 TMdomains, equivalent to a combination of theUT\u2010A2 and UT\u2010A3 splice variants.As a product of protein catabolism, urea is moved around the body and through the kidneys for excretion. Although there is experimental evidence for concentrative urea transporters, these have not been defined at the molecular level. The SLC14 family are facilitative transporters, allowing et al. (2015) Urea transporter proteins as targets for small\u2010molecule diuretics. Nat Rev Nephrol11: 113\u201023 [https://www.ncbi.nlm.nih.gov/pubmed/25488859?dopt=AbstractPlus]Esteva\u2010Font C et al. (2015) Evolution of urea transporters in vertebrates: adaptation to urea's multiple roles and metabolic sources. J. Exp. Biol. 218: 1936\u20101945 [https://www.ncbi.nlm.nih.gov/pubmed/26085670?dopt=AbstractPlus]LeMoine CM Am. J. Physiol. Regul. Integr. Comp. Physiol. 304: R488\u2010503 [https://www.ncbi.nlm.nih.gov/pubmed/23364530?dopt=AbstractPlus]Pannabecker TL. (2013) Comparative physiology and architecture associated with the mammalian urine concentrating mechanism: role of inner medullary water and urea transport pathways in the rodent medulla. et al. (2013) The urea transporter family (SLC14): physiological, pathological and structural aspects. Mol. Aspects Med. 34: 313\u201022 [https://www.ncbi.nlm.nih.gov/pubmed/23506873?dopt=AbstractPlus]Shayakul C Br. J. Pharmacol. 164: 1780\u201092 [https://www.ncbi.nlm.nih.gov/pubmed/21449978?dopt=AbstractPlus]Stewart G. (2011) The emerging physiological roles of the SLC14A family of urea transporters. +\u2010coupled oligopeptide cotransporter family, is a group of membrane transporters known for their key role in the cellular uptake of di\u2010 and tripeptides (di/tripeptides). Of its members, SLC15A1 (PEPT1) chiefly mediates intestinal absorption of luminal di/tripeptides from overall dietary protein digestion, SLC15A2 (PEPT2) mainly allows renal tubular reuptake of di/tripeptides from ultrafiltration and brain\u2010to\u2010blood efflux of di/tripeptides in the choroid plexus, SLC15A3 (PHT2) and SLC15A4 (PHT1) interact with both di/tripeptides and histidine, e.g. in certain immune cells, and SLC15A5 has unknown physiological function. In addition, the SLC15 family of peptide transporters variably interacts with a very large number of peptidomimetics and peptide\u2010like drugs. It is conceivable, based on the currently acknowledged structural and functional differences, to divide the SLC15 family of peptide transporters into two subfamilies.The Solute Carrier 15 (SLC15) family of peptide transporters, alias Hhttp://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4831, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4824, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4784, L\u2010Dopa prodrugs, gemcitabine prodrugs, floxuridine prodrugs, Maillard reaction products, JBP485, zanamivir, oseltamivir prodrugs, doxorubicin prodrugs, polymyxins, and didanosine prodrugs. Frequently used pharmaceutical excipients such as Tween\u00ae20, Tween\u00ae80, Solutol \u00aeHS 15 and Cremophor EL\u00aestrongly inhibit cellular uptake of Gly\u2010Sar by SLC15A1 and/or SLC15A2 [http://www.ncbi.nlm.nih.gov/pubmed/27903454?dopt=AbstractPlus]. There is evidence to suggest the existence of a fifth member of this transporter family, https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:33455 , but to date there is no established biological function or reported pharmacology for this protein [http://www.ncbi.nlm.nih.gov/pubmed/21044875?dopt=AbstractPlus].The members of the SLC15 family of peptide transporters are particularly promiscuous in the transport of di/tripeptides, and D\u2010amino acid containing peptides are also transported. While SLC15A3 and SLC15A4 transport histidine, none of them transport tetrapeptides. In addition, many molecules, among which beta\u2010lactam antibiotics, angiotensinconverting enzyme inhibitors and sartans, variably interact with the SLC15 family transporters. Known substrates include et al. (2010) Hijacking solute carriers for proton\u2010coupled drug transport. Physiology (Bethesda)25: 364\u201077 [https://www.ncbi.nlm.nih.gov/pubmed/21186281?dopt=AbstractPlus]Anderson CM Curr Opin Pharmacol13: 881\u20107 [https://www.ncbi.nlm.nih.gov/pubmed/24007794?dopt=AbstractPlus]Brandsch M. (2013) Drug transport via the intestinal peptide transporter PepT1. Expert Opin Drug Metab Toxicol5: 887\u2010905 [https://www.ncbi.nlm.nih.gov/pubmed/19519280?dopt=AbstractPlus]Brandsch M. (2009) Transport of drugs by proton\u2010coupled peptide transporters: pearls and pitfalls. et al. (2008) The solute carrier (SLC) complement of the human genome: phylogenetic classification reveals four major families. FEBS Lett. 582: 3811\u20106 [https://www.ncbi.nlm.nih.gov/pubmed/18948099?dopt=AbstractPlus]Fredriksson R Biochim. Biophys. Acta1850: 488\u2010499 [https://www.ncbi.nlm.nih.gov/pubmed/24859687?dopt=AbstractPlus]Newstead S. (2015) Molecular insights into proton coupled peptide transport in the PTR family of oligopeptide transporters. Curr. Opin. Struct. Biol. 45: 17\u201024 [https://www.ncbi.nlm.nih.gov/pubmed/27865112?dopt=AbstractPlus]Newstead S. (2017) Recent advances in understanding proton coupled peptide transport via the POT family. Biochem. Soc. Trans. 39: 1353\u20108 [https://www.ncbi.nlm.nih.gov/pubmed/21936814?dopt=AbstractPlus]Newstead S. (2011) Towards a structural understanding of drug and peptide transport within the proton\u2010dependent oligopeptide transporter (POT) family. et al. (2013) Proton\u2010coupled oligopeptide transporter family SLC15: physiological, pharmacological and pathological implications. Mol. Aspects Med. 34: 323\u201036 [https://www.ncbi.nlm.nih.gov/pubmed/23506874?dopt=AbstractPlus]Smith DE et al. (2007) H+\u2010coupled nutrient, micronutrient and drug transporters in the mammalian small intestine. Exp. Physiol. 92: 603\u201019 [https://www.ncbi.nlm.nih.gov/pubmed/17468205?dopt=AbstractPlus]Thwaites DT e.g. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2932), http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4809 and ketone bodies, as well as aromatic amino acids. Topology modelling suggests 12 TM domains, with intracellular termini and an extended loop at TM 6/7.Members of the SLC16 family may be divided into subfamilies on the basis of substrate selectivities, particularly lactate (e.g. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2932) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4809.The proton\u2010coupledmonocarboxylate transporters allowtransport of the products of cellularmetabolism, principally lactate Thyroid hormone transporters\u2010functions and clinical implications. Nat Rev Endocrinol11: 406\u2010417 https://www.ncbi.nlm.nih.gov/pubmed/25942657?dopt=AbstractPlusBernal J Mol. Aspects Med. 34: 337\u201049 https://www.ncbi.nlm.nih.gov/pubmed/23506875?dopt=AbstractPlusHalestrap AP. (2013) The SLC16 gene family \u2010 structure, role and regulation in health and disease. et al. (2016) Monocarboxylate Transporters: Therapeutic Targets and Prognostic Factors in Disease. Clin. Pharmacol. Ther. 100: 454\u2010463 https://www.ncbi.nlm.nih.gov/pubmed/27351344?dopt=AbstractPlusJones RS http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=221) and Type III (http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=195) transporters. Within the SLC17 family, however, further subgroups of organic anion transporters may be defined, allowing the accumulation of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4644 in the endoplasmic reticulum and glutamate (e.g. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1369) or nucleotides in synaptic and secretory vesicles. Topology modelling suggests 12 TM domains.The SLC17 family are sometimes referred to as Type I sodium\u2010phosphate co\u2010transporters, alongside Type II , an autosomal recessive neurodegenerative disorder associated with sialic acid storage disease [http://www.ncbi.nlm.nih.gov/pubmed/10581036?dopt=AbstractPlus].Loss\u2010of\u2010function mutations in sialin are associated with Salla disease (http://www.ncbi.nlm.nih.gov/pubmed/10938000?dopt=AbstractPlus].Vesicular glutamate transporters (VGLUTs) allow accumulation of glutamate into synaptic vesicles, as well as secretory vesicles in endocrine tissues. The roles of VGLUTs in kidney and liver are unclear. These transporters appear to utilize the proton gradient and also express a chloride conductance [http://www.ncbi.nlm.nih.gov/pubmed/20920794?dopt=AbstractPlus].Endogenous ketoacids produced during fasting have been proposed to regulate VGLUT function through blocking chloride ion\u2010mediated allosteric enhancement of transporter function .The vesicular nucleotide transporter is the most recent member of the SLC17 family to have an assigned function. Uptake of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4177 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4579.VGLUTs and VNUT can be inhibited by et al. (2017) Vesicular nucleotide transporter (VNUT): appearance of an actress on the stage of purinergic signaling. Purinergic Signal. 13: 387\u2010404 https://www.ncbi.nlm.nih.gov/pubmed/28616712?dopt=AbstractPlusMoriyama Y et al. (2016) Structure, Function, and Drug Interactions of Neurotransmitter Transporters in the Postgenomic Era. Annu. Rev. Pharmacol. Toxicol. 56: 385\u2010402 https://www.ncbi.nlm.nih.gov/pubmed/26514205?dopt=AbstractPlusOmote H Mol. Aspects Med. 34: 350\u20109 https://www.ncbi.nlm.nih.gov/pubmed/23506876?dopt=AbstractPlusReimer RJ. (2013) SLC17: a functionally diverse family of organic anion transporters. Pflugers Arch. 468: 513\u20108 https://www.ncbi.nlm.nih.gov/pubmed/26577586?dopt=AbstractPlusTakamori S. (2016) Vesicular glutamate transporters as anion channels? http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=137#V\u2010typeATPase that acidifies secretory vesicles (reviewed by [http://www.ncbi.nlm.nih.gov/pubmed/12827358?dopt=AbstractPlus]). The vesicular acetylcholine transporter localizes to cholinergic neurons, but non\u2010neuronal expression has also been claimed [http://www.ncbi.nlm.nih.gov/pubmed/21482687?dopt=AbstractPlus]. Vesicular monoamine transporter 1 is mainly expressed in peripheral neuroendocrine cells, but most likely not in the CNS, whereas VMAT2 [http://www.ncbi.nlm.nih.gov/pubmed/8643547?dopt=AbstractPlus] distributes between both central and peripheral sympathetic monoaminergic neurones [http://www.ncbi.nlm.nih.gov/pubmed/21272013?dopt=AbstractPlus]. The vescular polyamine transporter (VPAT) is highly expressed in the lungs and placenta, with moderate expression in brain and testis, and with low expression in heart and skeletal muscle [http://www.ncbi.nlm.nih.gov/pubmed/25355561?dopt=AbstractPlus]. VPAT mediates vesicular accumulation of polyamines in mast cells [http://www.ncbi.nlm.nih.gov/pubmed/28082679?dopt=AbstractPlus].The vesicular amine transporters (VATs) are putative 12 TM domain proteins that function to transport singly positively charged amine neurotransmitters and hormones from the cytoplasm and concentrate them within secretory vesicles. They function as amine/proton antiporters driven by secondary active transport utilizing the proton gradient established by a multi\u2010subunit pKi values for endogenous and synthetic substrate inhibitors of human VMAT1 and VMAT2 are for inhibition of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3248 uptake in transfected and permeabilised CV\u20101 cells as detailed by [http://www.ncbi.nlm.nih.gov/pubmed/8643547?dopt=AbstractPlus]. In addition to the monoamines listed in the table, the trace amines http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2150 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2144 are probable substrates for VMAT2 [http://www.ncbi.nlm.nih.gov/pubmed/21272013?dopt=AbstractPlus]. Probes listed in the table are those currently employed; additional agents have been synthesized (e.g. [http://www.ncbi.nlm.nih.gov/pubmed/19632829?dopt=AbstractPlus]).et al. (2015) Regulation of the Dopamine and Vesicular Monoamine Transporters: Pharmacological Targets and Implications for Disease. Pharmacol. Rev. 67: 1005\u201024 https://www.ncbi.nlm.nih.gov/pubmed/26408528?dopt=AbstractPlusGerman CL et al. (2017) Membrane transporters as mediators of synaptic dopamine dynamics: implications for disease. Eur. J. Neurosci. 45: 20\u201033 https://www.ncbi.nlm.nih.gov/pubmed/27520881?dopt=AbstractPlusLohr KM et al. (2016) Structure, Function, and Drug Interactions of Neurotransmitter Transporters in the Postgenomic Era. Annu. Rev. Pharmacol. Toxicol. 56: 385\u2010402 https://www.ncbi.nlm.nih.gov/pubmed/26514205?dopt=AbstractPlusOmote H et al. (2015) Amphetamines, new psychoactive drugs and the monoamine transporter cycle. Trends Pharmacol. Sci. 36: 41\u201050 https://www.ncbi.nlm.nih.gov/pubmed/25542076?dopt=AbstractPlusSitte HH Med Res Rev31: 483\u2010519 https://www.ncbi.nlm.nih.gov/pubmed/20135628?dopt=AbstractPlusWimalasena K. (2011) Vesicular monoamine transporters: structure\u2010function, pharmacology, and medicinal chemistry. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4563 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4629 are transported across the cell membrane, particularly in the intestine, kidneys and placenta, using pH differences as driving forces. Topological modelling suggests the transporters have 12 TM domains.The B vitamins http://www.ncbi.nlm.nih.gov/pubmed/10391223?dopt=AbstractPlus].Loss\u2010of\u2010function mutations in ThTr1 underlie thiamine\u2010responsive megaloblastic anemia syndrome .Mutations in the SLC22A5 gene lead to primary carnitine deficiency .URAT1, a member of the OAT (organic anion transporter) family, is an anion\u2010exchanging uptake transporter localized to the apical (brush border) membrane of renal proximal tubular cells. It is an anion exchanger that specifically reabsorbs uric acid from the proximal tubule in exchange for monovalent anions such as lactate, nicotinoate, acetoacetate, and hydroxybutyrate . The atypical SLCs share sequence similarities and phylogenetic ancestry with other SLCs, and they have historically been classified in to subfamilies ) based on phylogenetic, sequence and structural analyses [http://www.ncbi.nlm.nih.gov/pubmed/28878041?dopt=AbstractPlus].This family of transporters has previously been classified as part of the atypical major facilitator superfamily (MSF) protein superfamily and brivaracetam [http://www.ncbi.nlm.nih.gov/pubmed/26663401?dopt=AbstractPlus].There are three human synaptic vesicle glycoprotein 2 family members, SV2A, SV2B and SV2C. They have transmembrane transporter activity and can be classified in to the SLC superfamily of solute carriers in subfamily SLC22, as SCL22B1, B2 and B3 respectively. SV2A (SCL22B1) has been identified as the brain binding\u2010site for the antiepileptic drugs levetiracetam . Loss\u2010of\u2010function mutations in these genes are associated with hyperornithinemia\u2010hyperammonemia\u2010homocitrullinuria.Both ornithine transporters are inhibited by the polyamine http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1713 production.Mitochondrial phosphate transporters allow the import of inorganic phosphate for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1712 for mitochondrial http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1713. Further members of the mitochondrial nucleotide transporter subfamily convey diverse substrates including CoA, although not all members have had substrates identified.Mitochondrial nucleotide transporters, defined by structural similarlities, include the adenine nucleotide translocator family , which under conditions of aerobic metabolism, allow coupling between mitochondrial oxidative phosphorylation and cytosolic energy consumption by exchanging cytosolic Mitochondrial uncoupling proteins allow dissipation of the mitochondrial proton gradient associated with thermogenesis and regulation of radical formation.Many of the transporters identified below have yet to be assigned functions and are currently regarded as orphans.http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=209.Information on members of this family may be found in the Biochim. Biophys. Acta1858: 655\u2010664 https://www.ncbi.nlm.nih.gov/pubmed/28088333?dopt=AbstractPlusBaffy G. (2017) Mitochondrial uncoupling in cancer cells: Liabilities and opportunities. et al. (2017) UCP1: A transporter for H+ and fatty acid anions. Biochimie134: 28\u201034 https://www.ncbi.nlm.nih.gov/pubmed/27984203?dopt=AbstractPlusBertholet AM et al. (2013) The mitochondrial ADP/ATP carrier (SLC25 family): pathological implications of its dysfunction. Mol. Aspects Med. 34: 485\u201093 https://www.ncbi.nlm.nih.gov/pubmed/23506884?dopt=AbstractPlusCl\u00e9men\u00e7on B Mol. Aspects Med. 34: 465\u201084 https://www.ncbi.nlm.nih.gov/pubmed/23266187?dopt=AbstractPlusPalmieri F. (2013) The mitochondrial transporter family SLC25: identification, properties and physiopathology. et al. (2015) The mitochondrial phosphate carrier: Role in oxidative metabolism, calcium handling and mitochondrial disease. Biochem. Biophys. Res. Commun. 464: 369\u201075 https://www.ncbi.nlm.nih.gov/pubmed/26091567?dopt=AbstractPlusSeifert EL Trends Cell Biol. 27: 633\u2010644 https://www.ncbi.nlm.nih.gov/pubmed/28522206?dopt=AbstractPlusTaylor EB. (2017) Functional Properties of the Mitochondrial Carrier System. Along with the SLC4 family, the SLC26 family acts to allow movement of monovalent and divalent anions across cell membranes. The predicted topology is of 10\u201014 TM domains with intracellular C\u2010 and N\u2010termini, probably existing as dimers. Within the family, subgroups may be identified on the basis of functional differences, which appear to function as anion exchangers and anion channels (SLC26A7 and SLC26A9).et al. (2013) The SLC26 gene family of anion transporters and channels. Mol. Aspects Med. 34: 494\u2010515 https://www.ncbi.nlm.nih.gov/pubmed/23506885?dopt=AbstractPlusAlper SL et al. (2011) Regulation of electroneutral NaCl absorption by the small intestine. Annu. Rev. Physiol. 73: 261\u201081 https://www.ncbi.nlm.nih.gov/pubmed/21054167?dopt=AbstractPlusKato A et al. (2011) Pendrin function in airway epithelia. Cell. Physiol. Biochem. 28: 571\u20108 https://www.ncbi.nlm.nih.gov/pubmed/22116372?dopt=AbstractPlusNofziger C Kidney Int. 84: 657\u201066 https://www.ncbi.nlm.nih.gov/pubmed/23636174?dopt=AbstractPlusSoleimani M. (2013) SLC26 Cl\u2010/HCO3\u2010 exchangers in the kidney: roles in health and disease. http://www.ncbi.nlm.nih.gov/pubmed/11470793?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/7954810?dopt=AbstractPlus], transmembrane segments, and are predicted on the basis of structural similarities to form dimers. SLC27 members have several structural domains: integral membrane associated domain, peripheral membrane associated domain, FATP signature, intracellular AMP binding motif, dimerization domain, lipocalin motif, and an ER localization domain (identified in FATP4 only) . These transporters are unusual in that they appear to express intrinsic very longchain acyl\u2010CoA synthetase enzyme activity. Within the cell, these transporters may associate with plasma and peroxisomal membranes. FATP1\u20104 and \u20106 transport long\u2010 and very long\u2010chain fatty acids, while FATP5 transports long\u2010chain fatty acids as well as bile acids .Fatty acid transporter proteins (FATPs) are a family (SLC27) of six transporters (FATP1\u20106). They have at least one, and possibly six and FATP4 , as well as bile acid inhibitors of FATP5 [http://www.ncbi.nlm.nih.gov/pubmed/20448275?dopt=AbstractPlus], have been described; analysis of the mechanism of action of some of these inhibitors suggests that transport may be selectively inhibited without altering enzymatic activity of the FATP.Although the stoichiometry of fatty acid transport is unclear, it has been proposed to be facilitated by the coupling of fatty acid transport to conjugation with http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5496 accumulation has been used as a non\u2010selective index of fatty acid transporter activity.FATP2 has two variants: Variant 1 encodes the full\u2010length protein, while Variant 2 encodes a shorter isoform missing an internal protein segment. FATP6 also has two variants: Variant 2 encodes the same protein as Variant 1 but has an additional segment in the 5\u2032 UTR.et al. (2013) SLC27 fatty acid transport proteins. Mol. Aspects Med. 34: 516\u201328 https://www.ncbi.nlm.nih.gov/pubmed/23506886?dopt=AbstractPlusAnderson CM et al. (2015) Fatty acid transport proteins in disease: New insights from invertebrate models. Prog. Lipid Res. 60: 30\u201340 https://www.ncbi.nlm.nih.gov/pubmed/26416577?dopt=AbstractPlusDourlen P et al. (2010) Fatty acid transport across the cell membrane: regulation by fatty acid transporters. Prostaglandins Leukot. Essent. Fatty Acids82: 149\u201354 https://www.ncbi.nlm.nih.gov/pubmed/20206486?dopt=AbstractPlusSchwenk RW Nucleoside transporters are divided into two families, the sodium\u2010dependent, concentrative solute carrier family 28 (SLC28) and the equilibrative, solute carrier family 29 (SLC29). The endogenous substrates are typically nucleosides, although some family members can also transport nucleobases and organic cations.SLC28 family membersappear to have 13 TM segments with cytoplasmic N\u2010termini and extracellular C\u2010termini, and function as concentrative nucleoside transporters.et al. (2014) Structural basis of nucleoside and nucleoside drug selectivity by concentrative nucleoside transporters. Elife3: e03604 https://www.ncbi.nlm.nih.gov/pubmed/25082345?dopt=AbstractPlusJohnson ZL et al. (2008) SLC28 genes and concentrative nucleoside transporter (CNT) proteins. Xenobiotica38: 972\u201394 https://www.ncbi.nlm.nih.gov/pubmed/18668436?dopt=AbstractPlusPastor\u2010Anglada M et al. (2015) Nucleoside transporter proteins as biomarkers of drug responsiveness and drug targets. Front Pharmacol6: 13 https://www.ncbi.nlm.nih.gov/pubmed/25713533?dopt=AbstractPlusPastor\u2010Anglada M et al. (2018) Who Is Who in Adenosine Transport. Front Pharmacol9: 627 https://www.ncbi.nlm.nih.gov/pubmed/29962948?dopt=AbstractPlusPastor\u2010Anglada M et al. (2013) The human concentrative and equilibrative nucleoside transporter families, SLC28 and SLC29. Mol. Aspects Med. 34: 529\u201347 https://www.ncbi.nlm.nih.gov/pubmed/23506887?dopt=AbstractPlusYoung JD http://www.ncbi.nlm.nih.gov/pubmed/15701636?dopt=AbstractPlus]. ENT1\u20103 are described as broad\u2010spectrum equilibrative nucleoside transporters, while ENT4 is primarily a polyspecific organic cation transporter at neutral pH [http://www.ncbi.nlm.nih.gov/pubmed/21816955?dopt=AbstractPlus].SLC29 family members appear to be composed of 11 TM segments with cytoplasmic N\u2010termini and extracellular C\u2010termini. ENT1, ENT2 and ENT4 are cell\u2010surface transporters, while ENT3 is intracellular, possibly lysosomal , with subunits having six TM domains, and both termini being cytoplasmic. Dityrosine covalent linking has been suggested as a mechanism for dimerisation, particularly for ZnT3 [http://www.ncbi.nlm.nih.gov/pubmed/19521526?dopt=AbstractPlus]. The mechanism for zinc transport is unknown.Along with the http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=217.Information on members of this family may be found in the ZnT8/SLC30A8 is described as a type 1 diabetes susceptibility gene.Zinc fluxes may be monitored through the use of radioisotopic Zn\u201065 or the fluorescent dye FluoZin 3.et al. (2014) Contribution of calcium\u2010conducting channels to the transport of zinc ions. Pflugers Arch. 466: 381\u20137 https://www.ncbi.nlm.nih.gov/pubmed/23719866?dopt=AbstractPlusBouron A et al. (2016) Zinc transporters and signaling in physiology and pathogenesis. Arch. Biochem. Biophys. 611: 43\u201350 https://www.ncbi.nlm.nih.gov/pubmed/27394923?dopt=AbstractPlusHojyo S et al. (2013) The SLC30 family of zinc transporters \u2010 a review of current understanding of their biological and pathophysiological roles. Mol. Aspects Med. 34: 548\u201060 https://www.ncbi.nlm.nih.gov/pubmed/23506888?dopt=AbstractPlusHuang L et al. (2014) Current understanding of ZIP and ZnT zinc transporters in human health and diseases. Cell. Mol. Life Sci. 71: 3281\u201395 https://www.ncbi.nlm.nih.gov/pubmed/24710731?dopt=AbstractPlusKambe T et al. (2015) The Physiological, Biochemical, and Molecular Roles of Zinc Transporters in Zinc Homeostasis and Metabolism. Physiol. Rev. 95: 749\u2013784 https://www.ncbi.nlm.nih.gov/pubmed/26084690?dopt=AbstractPlusKambe T http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=138#Cu2+\u2010ATPase are involved in the regulation of cellular copper levels. The CTR1 transporter is a cell\u2010surface transporter to allow monovalent copper accumulation into cells, while CTR2 appears to be a vacuolar/vesicular transporter [http://www.ncbi.nlm.nih.gov/pubmed/15494390?dopt=AbstractPlus]. Functional copper transporters appear to be trimeric with each subunit having three TM regions and an extracellular N\u2010terminus. CTR1 is considered to be a higher affinity copper transporter compared to CTR2. The stoichiometry of copper accumulation is unclear, but appears to be energy\u2010independent [http://www.ncbi.nlm.nih.gov/pubmed/11734551?dopt=AbstractPlus].SLC31 family members, alongside the http://www.ncbi.nlm.nih.gov/pubmed/11734551?dopt=AbstractPlus].Copper accumulation through CTR1 is sensitive to silver ions, but not divalent cations , and is a member of the structurally\u2010defined amino acid\u2010polyamineorganocation/ APC clan composed of SLC32, SLC36 and SLC38 transporter families (see [http://www.ncbi.nlm.nih.gov/pubmed/23506890?dopt=AbstractPlus]). VIAAT was originally suggested to be composed of 10 TM segments with cytoplasmic N\u2010 and C\u2010termini [http://www.ncbi.nlm.nih.gov/pubmed/9349821?dopt=AbstractPlus]. However, an alternative 9TM structure with the N terminus facing the cytoplasm and the C terminus residing in the synaptic vesicle lumen has subsequently been reported [http://www.ncbi.nlm.nih.gov/pubmed/19052203?dopt=AbstractPlus]. VI\u2010AAT acts as an antiporter for inhibitory amino acids and protons. The accumulation ofGABA and glycine within vesicles is driven by both the chemical (\u0394pH) and electrical (\u0394\u03c8) components of the proton electrochemical gradient (\u0394\u03bcH+) established by a vacuolar H+\u2010ATPase [http://www.ncbi.nlm.nih.gov/pubmed/9349821?dopt=AbstractPlus]. However, one study, [http://www.ncbi.nlm.nih.gov/pubmed/19843525?dopt=AbstractPlus], presented evidence that VIAAT is instead a Cl\u2010/GABA co\u2010transporter. VIAAT co\u2010exists with http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=145#show_object_1007 (SLC17A7), or http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=145#show_object_1008 (SLC17A6), in the synaptic vesicles of selected nerve terminals . VIAAT knock out mice die between embryonic day 18.5 and birth [http://www.ncbi.nlm.nih.gov/pubmed/16701208?dopt=AbstractPlus]. In cultures of spinal cord neurones established from earlier embryos, the corelease of of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1067 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=727 from synaptic vesicles is drastically reduced, providing direct evidence for the role of VIAAT in the sequestration of both transmitters .The vesicular inhibitory amino acid transporter, VIAAT , which is the sole representative of the SLC32 family, transports et al. (2014) Vesicular neurotransmitter transporters: mechanistic aspects. Curr Top Membr73: 149\u201074 https://www.ncbi.nlm.nih.gov/pubmed/24745982?dopt=AbstractPlusAnne C et al. (2013) Evolutionary origin of amino acid transporter families SLC32, SLC36 and SLC38 and physiological, pathological and therapeutic aspects. Mol. Aspects Med. 34: 571\u201085 https://www.ncbi.nlm.nih.gov/pubmed/23506890?dopt=AbstractPlusSchi\u00f6th HB http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3038. SLC33A1/AT1 is a putative 11 TM transporter present on the endoplasmic reticulum, expressed in all tissues, but particularly abundant in the pancreas [http://www.ncbi.nlm.nih.gov/pubmed/9096318?dopt=AbstractPlus], which imports cytosolic http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3038 into these intracellular organelles.Acetylation of proteins is a post\u2010translational modification mediated by specific acetyltransferases, using the donor http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3038 transport through AT1 was inhibited by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3044, but not http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1058, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1713 or http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1782[http://www.ncbi.nlm.nih.gov/pubmed/20826464?dopt=AbstractPlus]. A loss\u2010of\u2010function mutation in ACATN1/SLC33A1 has been associated with spastic paraplegia , although this observation could not be replicated in a subsequent study [http://www.ncbi.nlm.nih.gov/pubmed/20461110?dopt=AbstractPlus].In heterologous expression studies, et al. (2004) The acetyl\u2010CoA transporter family SLC33. Pflugers Arch. 447: 760\u20102 https://www.ncbi.nlm.nih.gov/pubmed/12739170?dopt=AbstractPlusHirabayashi Y et al. (2013) The acetyl\u2010CoA transporter family SLC33. Mol. Aspects Med. 34: 586\u20109 https://www.ncbi.nlm.nih.gov/pubmed/23506891?dopt=AbstractPlusHirabayashi Y http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=145) and Type III (http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=195) transporters. Topological modelling suggests eight TM domains with C\u2010 and N\u2010 termini in the cytoplasm, and a re\u2010entrant loop at TM7/8. SLC34 family members are expressed on the apical surfaces of epithelia in the intestine and kidneys to regulate body phosphate levels, principally NaPi\u2010IIa and NaPi\u2010IIb, respectively. NaPi\u2010IIa and NaPi\u2010IIb are electrogenic, while NaPiIIc is electroneutral [http://www.ncbi.nlm.nih.gov/pubmed/18989094?dopt=AbstractPlus].The SLC34 family are sometimes referred to as Type II sodium\u2010phosphate co\u2010transporters, alongside Type I Phosphate transporters and their function. Annu. Rev. Physiol. 75: 535\u201050 https://www.ncbi.nlm.nih.gov/pubmed/23398154?dopt=AbstractPlusBiber J et al. (2013) Phosphate transporters of the SLC20 and SLC34 families. Mol. Aspects Med. 34: 386\u201095 https://www.ncbi.nlm.nih.gov/pubmed/23506879?dopt=AbstractPlusForster IC et al. (2014) Phosphate: an old bone molecule but new cardiovascular risk factor. Br J Clin Pharmacol77: 39\u201054 https://www.ncbi.nlm.nih.gov/pubmed/23506202?dopt=AbstractPlusShobeiri N et al. (2014) The SLC34 family of sodium\u2010dependent phosphate transporters. Pflugers Arch. 466: 139\u201053 https://www.ncbi.nlm.nih.gov/pubmed/24352629?dopt=AbstractPlusWagner CA Glycoprotein formation in the Golgi and endoplasmic reticulum relies on the accumulation of nucleotide\u2010conjugated sugars via the SLC35 family of transporters. These transporters have a predicted topology of 10 TM domains, with cytoplasmic termini, and function as exchangers, swopping nucleoside monophosphates for the corresponding nucleoside diphosphate conjugated sugar. Five subfamilies of transporters have been identified on the basis of sequence similarity, namely SLC35A1, SLC35A2, SLC35A3, SLC35A4 and SLC35A5; SLC35B1, SLC35B2, SLC35B3 and SLC35B4; SLC35C1 and SLC35C2; SLC35D1, SL35D1, SLC35D2 and SLC35D3, and the subfamily of orphan SLC35 transporters, SLC35E1\u20104 and SLC35F1\u20105.et al. (2004) Molecular physiology and pathology of the nucleotide sugar transporter family (SLC35). Pflugers Arch. 447: 768\u201075 https://www.ncbi.nlm.nih.gov/pubmed/12759756?dopt=AbstractPlusIshida N et al. (2016) Overview of Nucleotide Sugar Transporter Gene Family Functions Across Multiple Species. J. Mol. Biol. 428: 3150\u20103165 https://www.ncbi.nlm.nih.gov/pubmed/27261257?dopt=AbstractPlusOrellana A Mol. Aspects Med. 34: 590\u2010600 https://www.ncbi.nlm.nih.gov/pubmed/23506892?dopt=AbstractPlusSong Z. (2013) Roles of the nucleotide sugar transporters (SLC35 family) in health and disease. http://www.ncbi.nlm.nih.gov/pubmed/12761825?dopt=AbstractPlus, http://www.ncbi.nlm.nih.gov/pubmed/11390972?dopt=AbstractPlus]. PAT2 is expressed at the apical membrane of the renal proximal tubule [http://www.ncbi.nlm.nih.gov/pubmed/19033659?dopt=AbstractPlus] and at the plasma\u2010membrane in brown/beige adipocytes [http://www.ncbi.nlm.nih.gov/pubmed/25080478?dopt=AbstractPlus]. PAT1 and PAT4 are involved in regulation of the mTORC1 pathway [http://www.ncbi.nlm.nih.gov/pubmed/29971004?dopt=AbstractPlus]. More comprehensive lists of substrates can be found within the reviews under Further Reading and in the references.Members of the SLC36 family of proton\u2010coupled amino acid transporters are involved in membrane transport of amino acids and derivatives. The four transporters show variable tissue expression patterns and are expressed in various cell types at the plasma\u2010membrane and in intracellular organelles. PAT1 is expressed at the luminal surface of the small intestine and absorbs amino acids and derivatives [3]. In lysosomes, PAT1 functions as an effluxmechanism for amino acids produced during intralysosomal proteolysis .The SLC38 family of transporters appears to be responsible for the functionally\u2010defined system A and system N mechanisms of amino acid transport and are mostly expressed in the CNS. Two distinct subfamilies are identifiable within the SLC38 transporters. SNAT1, SNAT2 and SNAT4 appear to resemble system A transporters in accumulating neutral amino acids under the influence of the sodium gradient. SNAT3 and SNAT5 appear to resemble system N transporters in utilizing proton co\u2010transport to accumulate amino acids. The predicted membrane topology is of 11 TM domains with an extracellular C\u2010terminus and intracellular N\u2010terminus .Along with the http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4177 has been reported to inhibit cation accumulation through ZIP14 [http://www.ncbi.nlm.nih.gov/pubmed/18270315?dopt=AbstractPlus].Zinc fluxes may be monitored through the use of radioisotopic Zn\u201065 or the fluorescent dye FluoZin 3. The bicarbonate transport inhibitor et al. (2016) Zinc transporters and signaling in physiology and pathogenesis. Arch. Biochem. Biophys. 611: 43\u201350 https://www.ncbi.nlm.nih.gov/pubmed/27394923?dopt=AbstractPlusHojyo S et al. (2013) The SLC39 family of zinc transporters. Mol. Aspects Med. 34: 612\u20139 https://www.ncbi.nlm.nih.gov/pubmed/23506894?dopt=AbstractPlusJeong J et al. (2014) Current understanding of ZIP and ZnT zinc transporters in human health and diseases. Cell. Mol. Life Sci. 71: 3281\u201395 https://www.ncbi.nlm.nih.gov/pubmed/24710731?dopt=AbstractPlusKambe T et al. (2015) The Physiological, Biochemical, and Molecular Roles of Zinc Transporters in Zinc Homeostasis and Metabolism. Physiol. Rev. 95: 749\u2013784 https://www.ncbi.nlm.nih.gov/pubmed/26084690?dopt=AbstractPlusKambe T et al. (2014) Zinc: an underappreciated modulatory factor of brain function. Biochem. Pharmacol. 91: 426\u201335 https://www.ncbi.nlm.nih.gov/pubmed/25130547?dopt=AbstractPlusMarger L http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=183 of proton\u2010coupled metal transporters, ferroportin allows the accumulation of iron from the diet. Whilst SLC11A2 functions on the apical membrane, ferroportin acts on the basolateral side of the enterocyte, as well as regulating macrophage and placental iron levels. The predicted topology is of 12 TM domains, with intracellular termini [http://www.ncbi.nlm.nih.gov/pubmed/19150361?dopt=AbstractPlus], with the functional transporter potentially a dimeric arrangement . Ferroportin is essential for iron homeostasis [http://www.ncbi.nlm.nih.gov/pubmed/16054062?dopt=AbstractPlus]. Ferroportin is expressed on the surface of cells that store and transport iron, such as duodenal enterocytes, hepatocytes, adipocytes and reticuloendothelial macrophages. Levels of ferroportin are regulated by its association with (binding to) hepcidin, a 25 amino acid hormone responsive to circulating iron levels . Hepcidin binding targets ferroportin for internalisation and degradation, lowering the levels of iron export to the blood. Novel therapeutic agents which stabilise ferroportin or protect it from hepcidin\u2010induced degradation are being developed as antianemia agents. Anti\u2010ferroportin monoclonal antibodies are such an agent.Alongside the https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:15598, http://www.uniprot.org/uniprot/P81172), cleaved into http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5378 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5379 , is a small protein that increases upon inflammation, binds to ferroportin to regulate its cellular distribution and degradation. Gene disruption in mice results in embryonic lethality [http://www.ncbi.nlm.nih.gov/pubmed/16054062?dopt=AbstractPlus], while loss\u2010of\u2010function mutations in man are associated with haemochromatosis [http://www.ncbi.nlm.nih.gov/pubmed/15956209?dopt=AbstractPlus].Hepcidin The SLC40 basolateral iron transporter family (IREG1/ferroportin/MTP1). Pflugers Arch. 447: 801\u20136 https://www.ncbi.nlm.nih.gov/pubmed/12836025?dopt=AbstractPlusMcKie AT et al. (2013) Mammalian iron transporters: families SLC11 and SLC40. Mol. Aspects Med. 34: 270\u201387 https://www.ncbi.nlm.nih.gov/pubmed/23506870?dopt=AbstractPlusMontalbetti N 2+ efflux [http://www.ncbi.nlm.nih.gov/pubmed/22031603?dopt=AbstractPlus], possibly as a result of co\u2010expression of particular protein partners (see [http://www.ncbi.nlm.nih.gov/pubmed/23506895?dopt=AbstractPlus]). Topological modelling suggests 10 TM domains with cytoplasmic C\u2010 and N\u2010 termini.By analogy with bacterial orthologues, this family is probably magnesium transporters. The prokaryote orthologue, MgtE, is responsible for uptake of divalent cations, while the heterologous expression studies of mammalian proteins suggest Mget al. (2013) The structure and regulation of magnesium selective ion channels. Biochim. Biophys. Acta1828: 2778\u201092 https://www.ncbi.nlm.nih.gov/pubmed/23954807?dopt=AbstractPlusPayandeh J et al. (2013) The SLC41 family of MgtE\u2010like magnesium transporters. Mol. Aspects Med. 34: 620\u20108 https://www.ncbi.nlm.nih.gov/pubmed/23506895?dopt=AbstractPlusSahni J et al. (2014) SLC41 transporters\u2013molecular identification and functional role. Curr Top Membr73: 383\u2010410 https://www.ncbi.nlm.nih.gov/pubmed/24745990?dopt=AbstractPlusSchweigel\u2010R\u00f6ntgen M https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:10008 (http://www.uniprot.org/uniprot/P18577) and https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:10009 (http://www.uniprot.org/uniprot/Q02161), expressed on the surface of erythrocytes. On erythrocytes, RhAG associates with these antigens and functions as an ammonium transporter. RhBG and RhBG are non\u2010erythroid related sequences associated with epithelia. Topological modelling suggests the presence of 12TM with cytoplasmic N\u2010 and C\u2010 termini. The majority of information on these transporters derives from orthologues in yeast, plants and bacteria. More recent evidence points to family members being permeable to carbon dioxide, leading to the term gas channels.Rhesus is commonly defined as a \u2019factor\u2019 that determines, in part, blood type, and whether neonates suffer from haemolytic disease of the newborn. These glycoprotein antigens derive from two genes, et al. (2013) Characteristics of mammalian Rh glycoproteins (SLC42 transporters) and their role in acid\u2010base transport. Mol. Aspects Med. 34: 629\u201037 https://www.ncbi.nlm.nih.gov/pubmed/23506896?dopt=AbstractPlusNakhoul NL et al. (2011) Role of NH3 and NH4+ transporters in renal acid\u2010base transport. Am. J. Physiol. Renal Physiol. 300: F11\u201023 https://www.ncbi.nlm.nih.gov/pubmed/21048022?dopt=AbstractPlusWeiner ID et al. (2014) Ammonia transport in the kidney by Rhesus glycoproteins. Am. J. Physiol. Renal Physiol. 306: F1107\u201020 https://www.ncbi.nlm.nih.gov/pubmed/24647713?dopt=AbstractPlusWeiner ID http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=141#SLC7 family. LAT3 and LAT4 contain 12 put.ative TM domains with both N and C termini located intracellularly. They transport neutral amino acids in a manner independent of Na+ and Cl\u2010 and with two kinetic components . LAT3/SLC43A1 is expressed in human tissues at high levels in the pancreas, liver, skeletal muscle and fetal liver [http://www.ncbi.nlm.nih.gov/pubmed/12930836?dopt=AbstractPlus] whereas LAT4/SLC43A2 is primarily expressed in the placenta, kidney and peripheral blood leukocytes [http://www.ncbi.nlm.nih.gov/pubmed/15659399?dopt=AbstractPlus]. SLC43A3 is expressed in vascular endothelial cells [http://www.ncbi.nlm.nih.gov/pubmed/18483404?dopt=AbstractPlus] but remains to be characterised.LAT3 (SLC43A1) and LAT4 (SLC43A2) are transporters with system L amino acid transporter activity, along with the structurally and functionally distinct transporters LAT1 and LAT2 that are members of the http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5335 inhibits its function [http://www.ncbi.nlm.nih.gov/pubmed/12930836?dopt=AbstractPlus] and at LAT4 inhibits the low\u2010, but not high\u2010affinity component of transport [http://www.ncbi.nlm.nih.gov/pubmed/15659399?dopt=AbstractPlus].Covalent modification of LAT3 by et al. (2013) The small SLC43 family: facilitator system l amino acid transporters and the orphan EEG1. Mol. Aspects Med. 34: 638\u201045 https://www.ncbi.nlm.nih.gov/pubmed/23268354?dopt=AbstractPlusBodoy S http://www.ncbi.nlm.nih.gov/pubmed/15715662?dopt=AbstractPlus]. CTL family members are putative 10TM domain proteins with extracellular termini that mediate Na+\u2010independent transport of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4551 with an affinity that is intermediate to that of the high affinity choline transporter CHT1 (http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=143#show_object_914) and the low affinity organiccation transporters [OCT1 (http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=146#show_object_1019) andOCT2 (http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=146#show_object_1020)] [http://www.ncbi.nlm.nih.gov/pubmed/16636297?dopt=AbstractPlus]. CLT1 is expressed almost ubiquitously in human tissues [http://www.ncbi.nlm.nih.gov/pubmed/11698453?dopt=AbstractPlus] and mediates http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4551 transport across the plasma and mitochondrial membranes [http://www.ncbi.nlm.nih.gov/pubmed/19357133?dopt=AbstractPlus]. Transport of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4551 by CTL2, which in rodents is expressed as two isoforms in lung, colon, inner ear and spleen and to a lesser extent in brain, tongue, liver, and kidney, has only recently been demonstrated . CTL3\u20105 remain to be characterized functionally.Members of the choline transporter\u2010like family are encoded by five genes (CTL1\u2010CTL5) with further diversity occurring through alternative splicing of CTL1, 4 and 5 [http://www.ncbi.nlm.nih.gov/pubmed/16000150?dopt=AbstractPlus]; rat renal tubule epithelial cells [http://www.ncbi.nlm.nih.gov/pubmed/19236841?dopt=AbstractPlus]; human colon carcinoma cells [http://www.ncbi.nlm.nih.gov/pubmed/19135976?dopt=AbstractPlus]; human keratinocytes [http://www.ncbi.nlm.nih.gov/pubmed/19122366?dopt=AbstractPlus] and human neuroblastoma cells [http://www.ncbi.nlm.nih.gov/pubmed/21185344?dopt=AbstractPlus]. Choline uptake by CLT1 is inhibited by numerous organic cations . In the guinea\u2010pig, CTL2 is a target for antibody\u2010induced hearing loss [http://www.ncbi.nlm.nih.gov/pubmed/14973250?dopt=AbstractPlus] and in man, a polymorphism in CTL2 constitutes the human neutrophil alloantigen\u20103a .Data tabulated are features observed for CLT1 endogenous to: rat astrocytes [Biopharm Drug Dispos35: 431\u201049 https://www.ncbi.nlm.nih.gov/pubmed/24532461?dopt=AbstractPlusInazu M. (2014) Choline transporter\u2010like proteins CTLs/SLC44 family as a novel molecular target for cancer therapy. et al. (2013) The choline transporter\u2010like family SLC44: properties and roles in human diseases. Mol. Aspects Med. 34: 646\u201054 https://www.ncbi.nlm.nih.gov/pubmed/23506897?dopt=AbstractPlusTraiffort E http://www.ncbi.nlm.nih.gov/pubmed/12417639?dopt=AbstractPlus]. The protein is predicted to have 12TM domains, with intracellular termini. The SLC45A2 gene is thought to encode a transporter protein that mediates http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5415 synthesis. Mutations in SLC45A2 are a cause of oculocutaneous albinism type 4 (e.g. [http://www.ncbi.nlm.nih.gov/pubmed/11574907?dopt=AbstractPlus]), and polymorphisms in this gene are associated with variations in skin and hair color (e.g. [http://www.ncbi.nlm.nih.gov/pubmed/15714523?dopt=AbstractPlus]).Members of the SLC45 family remain to be fully characterised. SLC45A1 was initially identified in the rat brain, particularly predominant in the hindbrain, as a proton\u2010associated sugar transport, induced by hypercapnia [et al. (2014) Proton\u2010associated sucrose transport of mammalian solute carrier family 45: an analysis in Saccharomyces cerevisiae. Biochem. J. 464: 193\u2010201 https://www.ncbi.nlm.nih.gov/pubmed/25164149?dopt=AbstractPlusBart\u00f6lke R et al. (2013) The SLC45 gene family of putative sugar transporters. Mol. Aspects Med. 34: 655\u201060 https://www.ncbi.nlm.nih.gov/pubmed/23506898?dopt=AbstractPlusVitavska O http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4563 transporter [http://www.ncbi.nlm.nih.gov/pubmed/17129779?dopt=AbstractPlus], with lower affinity for http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4349. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4563 accumulation is independent of Na+ or K+ ion concentrations, but driven by extracellular protons with an as yet undefined stoichiometry.Based on the proptypicalmember of this family, PCFT, this family includes proton\u2010driven transporters with 11 TMsegments. SLC46A1 has been described to act as an intestinal proton\u2010coupled high\u2010affinity et al. (2014) Biology of the major facilitative folate transporters SLC19A1 and SLC46A1. Curr Top Membr73: 175\u2010204 https://www.ncbi.nlm.nih.gov/pubmed/24745983?dopt=AbstractPlusHou Z et al. (2014) The major facilitative folate transporters solute carrier 19A1 and solute carrier 46A1: biology and role in antifolate chemotherapy of cancer. Drug Metab. Dispos. 42: 632\u201049 https://www.ncbi.nlm.nih.gov/pubmed/24396145?dopt=AbstractPlusMatherly LH et al. (2015) Structural determinants of human proton\u2010coupled folate transporter oligomerization: role of GXXXG motifs and identification of oligomeric interfaces at transmembrane domains 3 and 6. Biochem. J. 469: 33\u201044 https://www.ncbi.nlm.nih.gov/pubmed/25877470?dopt=AbstractPlusWilson MR et al. (2011) Mechanisms of membrane transport of folates into cells and across epithelia. Annu. Rev. Nutr. 31: 177\u2010201 https://www.ncbi.nlm.nih.gov/pubmed/21568705?dopt=AbstractPlusZhao R et al. (2013) Folate and thiamine transporters mediated by facilitative carriers (SLC19A1\u20103 and SLC46A1) and folate receptors. Mol. Aspects Med. 34: 373\u201085 https://www.ncbi.nlm.nih.gov/pubmed/23506878?dopt=AbstractPlusZhao R http://www.ncbi.nlm.nih.gov/pubmed/19515813?dopt=AbstractPlus] and are suggested to be responsible for excretion of many drugs in the liver and kidneys.These proton:organic cation exchangers are predicted to have 13 TM segments [http://www.ncbi.nlm.nih.gov/pubmed/20047987?dopt=AbstractPlus]. MATE2 and MATE2\u2010B are inactive splice variants of MATE2\u2010K [http://www.ncbi.nlm.nih.gov/pubmed/16807400?dopt=AbstractPlus].DAPI has been used to allow quantification of MATE1 and MATE2\u2010mediated transport activity . In addition, evidence suggests this 4TM\u2010containing protein associates with the http://www.guidetopharmacology.org/GRAC/FamilyDisplayForward?familyId=137#V\u2010type ATPase in lysosomes [http://www.ncbi.nlm.nih.gov/pubmed/19875448?dopt=AbstractPlus]. Recent studies confirm its lysosomal location and demonstrate that it has an important physiological function in macrophages ingesting senescent red blood cells (erythrophagocytosis), recycling heme (released from the red cell hemoglobin) from the phagolysosome into the cytosol, where the heme is subsequently catabolized to recycle the iron [http://www.ncbi.nlm.nih.gov/pubmed/23395172?dopt=AbstractPlus].HRG1 has been identified as a cell surface and lysosomal heme transporter [et al. (2013) Heme and FLVCR\u2010related transporter families SLC48 and SLC49. Mol. Aspects Med. 34: 669\u201082 https://www.ncbi.nlm.nih.gov/pubmed/23506900?dopt=AbstractPlusKhan AA http://www.ncbi.nlm.nih.gov/pubmed/10400745?dopt=AbstractPlus], and later identified as a cell surface accumulation which exports heme from the cytosol [http://www.ncbi.nlm.nih.gov/pubmed/15369674?dopt=AbstractPlus]. A recent study indicates that an isoform of FLVCR1 is located in the mitochondria, the site of the final steps of heme synthesis, and appears to transport heme into the cytosol [http://www.ncbi.nlm.nih.gov/pubmed/23187127?dopt=AbstractPlus]. FLVCR\u2010mediated heme transport is essential for erythropoiesis. Flvcr1 gene mutations have been identified as the cause of PCARP (http://www.omim.org/entry/609033?search=609033&highlight=609033 (PCARP) [http://www.ncbi.nlm.nih.gov/pubmed/21070897?dopt=AbstractPlus].There are three paralogs of FLVCR1 in the human genome.FLVCR1 was initially identified as a cell\u2010surface attachment site for feline leukemia virus subgroup C [http://www.ncbi.nlm.nih.gov/pubmed/11943475?dopt=AbstractPlus], has been reported to function as a heme importer [http://www.ncbi.nlm.nih.gov/pubmed/20823265?dopt=AbstractPlus]. In addition, a congenital syndrome of proliferative vasculopathy and hydranencephaly, also known as Fowler's syndrome, is associated with a loss\u2010of\u2010function mutation in FLVCR2 [http://www.ncbi.nlm.nih.gov/pubmed/20206334?dopt=AbstractPlus].FLVCR2, most similar to FLVCR1 [http://www.ncbi.nlm.nih.gov/pubmed/11912179?dopt=AbstractPlus].The functions of the other two members of the SLC49 family, MFSD7 and DIRC2, are unknown, although DIRC2 has been implicated in hereditary renal carcinomas [http://omim.org/entry/105650 [http://www.ncbi.nlm.nih.gov/pubmed/18815190?dopt=AbstractPlus].Non\u2010functional splice alternatives of FLVCR1 have been implicated as a cause of a congenital red cell aplasia, et al. (2013) Heme and FLVCR\u2010related transporter families SLC48 and SLC49. Mol. Aspects Med. 34: 669\u201082 https://www.ncbi.nlm.nih.gov/pubmed/23506900?dopt=AbstractPlusKhan AA et al. (2011) Control of intracellular heme levels: heme transporters and heme oxygenases. Biochim. Biophys. Acta1813: 668\u201082 https://www.ncbi.nlm.nih.gov/pubmed/21238504?dopt=AbstractPlusKhan AA http://www.ncbi.nlm.nih.gov/pubmed/8630032?dopt=AbstractPlus], later termed Rag1\u2010activating protein 1, with a sequence homology predictive of a 4TM topology. The plant orthologues, termed SWEETs, appear to be 7 TM proteins, with extracellular N\u2010termini, and the capacity for bidirectional flux of http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4536 [http://www.ncbi.nlm.nih.gov/pubmed/21107422?dopt=AbstractPlus]. Expression of mouse SWEET in the mammary gland was suggestive of a role in Golgi lactose synthesis [http://www.ncbi.nlm.nih.gov/pubmed/21107422?dopt=AbstractPlus].A mouse stromal cell cDNA library was used to clone C2.3 . OST\u03b1/OST\u03b2 is also expressed in steroidogenic cells of the brain and adrenal gland, where it may contribute to steroid movement [http://www.ncbi.nlm.nih.gov/pubmed/20649839?dopt=AbstractPlus]. Bile acid transport is suggested to be facilitative and independent of sodium, potassium, chloride ions or protons . OST\u03b1/OST\u03b2 heterodimers have been shown to transport http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4546, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5577, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=4748, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=6504 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5577 . OST\u03b1/OST\u03b2\u2010mediated transport of bile salts is inhibited by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=9184 [http://www.ncbi.nlm.nih.gov/pubmed/29675448?dopt=AbstractPlus]. OST\u03b1 is suggested to be a seven TM protein, while OST\u03b2 is a single TM \u2018ancillary\u2019 protein, both of which are thought to have intracellular C\u2010termini [http://www.ncbi.nlm.nih.gov/pubmed/17650074?dopt=AbstractPlus]. Both proteins function in solute transport and bimolecular fluorescence complementation studies suggest the possibility of OST\u03b1 homooligomers, as well as OST\u03b1/OST\u03b2 hetero\u2010oligomers . An inherited mutation in OST\u03b2 is associated with congenital diarrhea in children [http://www.ncbi.nlm.nih.gov/pubmed/28898457?dopt=AbstractPlus].The SLC51 organic solute transporter family of transporters is a pair of heterodimeric proteins which regulate bile salt movements in the small intestine, bile duct, and liver, as part of the enterohepatic circulation Yonezawa A SLC54 family transporters appear to function as mechanisms for accumulating pyruvate into mitochondria to link glycolysis with oxidative phosphorylation.The family of SLC55 mitochondrial transporters appear to regulate ion fluxes and to maintain tubular networks.These are a family of incompletely\u2010characterised mitochondrial transporters.The SLC63 family of transporters has roles inside the cell (SLC63A1/SPNS1) or on the cell surface (SLC63A2/SPNS2) in sphingolipid transport.https://www.uniprot.org/uniprot/P61916), allows the accumulation into the cytosol of cholesterol acquired from low density lipoproteins.The SLC65 family of intracellular cholesterol transporters are 13TM membrane proteins. NPC1/SLC65A1 is an intracellular cholesterol transporter, which together with NPC2 (Uniprot ID The SLCO superfamily is comprised of the organic anion transporting polypeptides (OATPs). The 11 human OATPs are divided into 6 families and ten subfamilies based on amino acid identity. These proteins are located on the plasma membrane of cells throughout the body. They have 12 TM domains and intracellular termini, with multiple putative glycosylation sites. OATPs mediate the sodium\u2010independent uptake of a wide range of amphiphilic substrates, including many drugs and toxins. Due to the multispecificity of these proteins, this guide lists classes of substrates and inhibitors for each family member. More comprehensive lists of substrates, inhibitors, and their relative affinities may be found in the review articles listed below.et al. (2013) The SLCO (former SLC21) superfamily of transporters. Mol. Aspects Med. 34: 396\u2010412 https://www.ncbi.nlm.nih.gov/pubmed/23506880?dopt=AbstractPlusHagenbuch B et al. (2013) Emerging transporters of clinical importance: an update from the International Transporter Consortium. Clin. Pharmacol. Ther. 94: 52\u201063 https://www.ncbi.nlm.nih.gov/pubmed/23588305?dopt=AbstractPlusHillgren KM et al. (2010) Membrane transporters in drug development. Nat Rev Drug Discov9: 215\u201036 https://www.ncbi.nlm.nih.gov/pubmed/20190787?dopt=AbstractPlusInternational Transporter Consortium et al. (2017) Interindividual and interethnic variability in drug disposition: polymorphisms in organic anion transporting polypeptide 1B1 . Br J Clin Pharmacol83: 1176\u20101184 https://www.ncbi.nlm.nih.gov/pubmed/27936281?dopt=AbstractPlusLee HH et al. (2012) OATPs, OATs and OCTs: the organic anion and cation transporters of the SLCO and SLC22A gene superfamilies. Br. J. Pharmacol. 165: 1260\u201087 https://www.ncbi.nlm.nih.gov/pubmed/22013971?dopt=AbstractPlusRoth M et al. (2018) Transporters in Drug Development: 2018 ITC Recommendations for Transporters of Emerging Clinical Importance. Clin. Pharmacol. Ther. 104: 890\u2010899 https://www.ncbi.nlm.nih.gov/pubmed/30091177?dopt=AbstractPlusZamek\u2010Gliszczynski MJ et al. (2016) SLC transporters as a novel class of tumour suppressors: identity, function and molecular mechanisms. Biochem. J. 473: 1113\u201024 https://www.ncbi.nlm.nih.gov/pubmed/27118869?dopt=AbstractPlusBhutia YD et al. (2016) SLC Transporters: Structure, Function, and Drug Discovery. Medchemcomm7: 1069\u20101081 https://www.ncbi.nlm.nih.gov/pubmed/27672436?dopt=AbstractPlusColas C et al. (2015) A Call for Systematic Research on Solute Carriers. Cell162: 478\u201087 https://www.ncbi.nlm.nih.gov/pubmed/26232220?dopt=AbstractPlusC\u00e9sar\u2010Razquin A et al. (2015) SLC transporters as therapeutic targets: emerging opportunities. Nat Rev Drug Discov14: 543\u201060 https://www.ncbi.nlm.nih.gov/pubmed/26111766?dopt=AbstractPlusLin L a|cz KA. (2017) Solute Carriers in the Blood\u2010Brain Barier: Safety in Abundance. Neurochem. Res. 42: 795\u2010809 https://www.ncbi.nlm.nih.gov/pubmed/27503090?dopt=AbstractPlusNaet al. (2016) Impact of Membrane Drug Transporters on Resistance to Small\u2010Molecule Tyrosine Kinase Inhibitors. Trends Pharmacol. Sci. 37: 904\u2010932 https://www.ncbi.nlm.nih.gov/pubmed/27659854?dopt=AbstractPlusNeul C Nat Rev Drug Discov14: 29\u201044 https://www.ncbi.nlm.nih.gov/pubmed/25475361?dopt=AbstractPlusNigam SK. (2015) What do drug transporters really do? et al. (2016) Glycosylation of solute carriers: mechanisms and functional consequences. Pflugers Arch. 468: 159\u201076 https://www.ncbi.nlm.nih.gov/pubmed/26383868?dopt=AbstractPlusPedersen NB et al. (2017) Classification Systems of Secondary Active Transporters. Trends Pharmacol. Sci. 38: 305\u2010315 https://www.ncbi.nlm.nih.gov/pubmed/27939446?dopt=AbstractPlusPerland E et al. (2017) Potentiating SLC transporter activity: Emerging drug discovery opportunities. Biochem. Pharmacol. 135: 1\u201011 https://www.ncbi.nlm.nih.gov/pubmed/28214518?dopt=AbstractPlusRives ML"} +{"text": "Without reviewers dedicating their expertise and time to supporting the timely peer review of these studies, we would not be successful in creating a venue that publishes the highest-quality scientific literature. Solid peer review creates trust that the work published in mSystems is reliable. However, it is also fun to remember that like pedestrians, cyclists, and motorists, who can all switch places at some point and yet fail to act with sufficient empathy toward the other two, reviewers, authors, and editors of society journals often need to remember that the service that they provide will reflect on the service that they receive. So thank you to all the reviewers for doing unto others as you would have done unto thyself! Thank you also for supporting open access publishing, and we look forward to exploring the power of open peer review in 2020.Frank M. AarestrupSophie Saphia AbbyMark D. AdamsJoshua N. AdkinsAnton AebischerAmeeta K. AgarwalMasato AkibaAlexander AksenovLuis D. AlcarazNezar Al-HebshiCaitilyn AllenJacob AllenEmma Allen-VercoeJo\u00e3o M. G. C. F. AlmeidaHasan Ali Al-TalhiStefano AmalfitanoKatherine R. AmatoNamasivayam AmbalavananAmnon AmirAlongkorn AmnuaykanjanasinKarthik AnantharamanTessa AndermanBrooke AndersonChristopher Joseph AndersonRika AndersonAndr\u00e9s Andrade-Dom\u00ednguezHaike AntelmannJosefa Ant\u00f3nRafael AraosAna Paula ArezHector ArguelloAnna Ar\u00edsE. Virginia ArmbrustKristine ArnvigMario Arrieta-OrtizBernard P. ArulanandamAlexander AskenovMarina Elisabeth AspholmLouis AtesFrank O. AylwardRamy K. AzizGiovanni BacciBrett J. BakerKatherine BarbeauSonia L. BardyMohammed BarigouTyler BarnumLars BarquistDariusz BartosikJose M. BautistaBarbara BayerMichael BaymWendy BedaleVivian BellofattoRonen Ben-AmiSarah Ben MaamarRichard J. BennettKathryn BernardHans C. BernsteinClaire BertelliStefan BertilssonCora BetsingerRodrigo Carvalho BicalhoSteven BillerEmanuele G. BiondiWilbert BitterLinda BlackallJeffrey BlanchardJill R. BlankenshipLouis-Marie BobayRich BodenNicholas Andrew BokulichJennifer M. BombergerElizaveta A. Bonch-OsmolovskayaJoe Bondy-DenomyRafaella C. Bonugli-SantosEmanuele BosiGerm\u00e1n BouSebastien BoutinRobert BowersJohn Dallas BoyceNanette R. BoyleJoel BozueGerrit BrandisKristoffer BrandvoldAsker Daniel BrejnrodPatrizia BrigidiIlana Lauren BritoRobert A. BrittonNichole A. BroderickDennis BrownPamela J. B. BrownSilvio D. BruggerJennifer BrumDonald A. BryantSamuel BrysonDongbo BuAlison BuchanSilvia BulgheresiLorinda BullingtonJoy BuongiornoMariana X. ByndlossL\u00e9a CabrolMichelle C. CalleganAntonio CamargoValerie Jean CarabettaKyle CardMaureen A. CareyErin CarlsonAlex CarrStephanie A. CarrV\u00edctor J. CarrionEric CascalesGrayson L. ChadwickBaofeng ChaiJosephine R. ChandlerYanjie ChaoTrevor CharlesAleksandra ChecinskaCasey ChenLiang ChenTingtao ChenWei ChenXingqun ChengYin-Ru ChiangSiok-Fong ChinBarbara ChirulloLudmila ChistoserdovaByung-Kwan ChoHana Cipcic PaljetakJan ClaesenC\u00e9cile ClavaudDavid W. ClearyLuis Pedro CoelhoMaureen ColemanJames CollinsFergus CollinsAndre M. ComeauLydia M. ContrerasBrian K. CoombesKevin M. CoombsVaughn S. CooperTeresa M. CoqueJacques CorbeilMelissa A. CreggerJohn CryanChristina A. CuomoTom CurtisWashington da SilvaJos\u00e9 Freire da Silva NetoEmily DavenportAlan R. DavidsonThomas DawsonSvetlana N. DedyshClaire de La SerreManuel Delgado-BaquerizoLaurence DelhaesElaine Cristina De MartinisAlexandre de MenezesMarjan De MeyHaiteng DengMahesh DesaiMatsapume DetcharoenSuzanne DevkotaWillem M. de VosBert DevriendtFloyd E. DewhirstGeorge C. DicenzoChristian DienerC\u00e9sar D\u00edez-Villase\u00f1orPatricia A. Digiuseppe ChampionJoseph P. DillardCecilia Di RubertoDaniel DistelDirk P. DittmerYohei DoiTao G. DongAnn DonoghueRodolpho Martin do PradoTobias D\u00f6rrGavin M. DouglasSimon L. DoveTheo W. DreherAdam DriksAlexandre DrouinEdward G. DudleyJohn DunbarSylvia DuncanClaire DuvalletMohammed DwidarJoseph EdwardsTimothy Ejike EgboPatrick EichenbergerMark A. EitemanNazira El-HageFadi Elias El-RamiBert ElyPhilipp EngelWhitney Eileen EnglandCarmen EspejoA. V. Espinel-IngroffMehrbod EstakiAlessandra S. EustaquioEdouard EvangelistiJoseph O. FalkinhamKaroline FaustConor FeehilyGabriel da Rocha FernandesCelio Fernando Figueiredo AngoliniScott G. FillerSteven E. FinkelMarco FondiAdriana ForeroLeonard J. FosterFarnaz FouladiMichael FranceKurt FredrickMarcelo FreireSteven FreseMichael W. FriedrichBettina C. FriesJulia FukuyamaOhad Gal-MorAlexander GamischMichael G. GanzleFrancisco Garc\u00eda-Del PortilloNeha GargRoger GarrettH. R. GaskinsJosep M. GasolKristi GdanetzMikhail S. GelfandRaad GharaibehScott Michael GiffordDouglas Paul GladueLaura GlendinningGregory B. GloorErica M. GossBenjamin GoudeyRevathi GovindDavid C. GraingerLeigh GreathouseTodd M. GrecoKacy GreenhalghAnn GregoryHans J. GriesserRobert GriffinsMaureen GroerMathieu GroussinHarald R. Gruber-VodickaCalin GuetBrian HaasElaine M. HaaseMike HadfieldLive H. HagenMatthias HahnJarrad Hampton-MarcellKim M. HandleyCara H. HaneyWilliam R. HarcombeDennis J. Hartigan-O\u2019ConnorMohamed-Amine HassaniStijn HawinkelRichard HayesAnna Heintz-BuschartBernard HenrissatChris HenryMarkus J. Herrg\u00e5rdDeborah M. HintonKendal HirschiThomas HitchMengfei HoLucas R. HoffmanDeborah A. HoganGeorgina Louise HoldNicola HoldenHsin-Ho HuangJean J. HuangShi HuangYi-Xin HuoRobert HutkinsMuhammad ImranIkbal Agah InceMarie-Agn\u00e8s JacquesPratik JagtapLaura R. JarboeMichael JewettLin JiangShuo JiaoJay-Hyun JoRheinallt M. JonesSusan JosephSean P. JungbluthFatah KashanchiPurna Chandra KashyapJustin R. KasparKevin T. KavanaghDaniel B. KearnsScott P. KeelyScott T. KelleyEduard J. KerkhovenMegan R. KiedrowskiMinsuk KimPan-Jun KimSeon-Won KimBenjamin KingKuniki KinoNichole R. KlattVanja Klepac-CerajMartin G. KlotzClaudia KniefDan KnightsMatthew D. KociUma KoduruArash KomeiliHeidi KongKonstantinos T. KonstantinidisOmry KorenPanagiotis KougiasElizabeth B. KujawinskiPrashant KumarRoshan KumarRanjith KumavathBenoit KunathThomas KuyperLeo LahtiChad R. LaingCalvin Ho-Fung LauAdi LavySarah LebeerSarah L. LebeisKyongbum LeeGabriela G. S. LeiteVanessa LeoneElisabeth LetellierMark Alexander LeverRoger C. LevesqueDavid Levy-BoothRuth E. LeyHuiying LiXian-Zhi LiXin-Di LiaoXiaoxia LinMary LiptonJohn J. LipumaJinhua LiuJinxin LiuYubing LiuKenneth J. LoceyFrank E. L\u00f6fflerTalita Louren\u00e7oJames LoweTiffany Marie Lowe-PowerSebastian LueckerTillmann LuedersDiana LuisePeter Adrian LundAntoni LuqueCourtney LuterbachMichael LyuWenjun MaFrank Michael MaixnerThulani P. MakhalanyaneAntoine MalabiradeAntonino Malacrin\u00f3Anthony MalanoskiRex R. MalmstromAshutosh K. MangalamaSam MannaFei MaoMaria L. MarcoChristopher W. MarshallPhilippe MarteauWillm Martens-HabbenaJose-Luis Martinez-GonzalezFumito MaruyamaNorman MauderMeghan MayAndrew J. McbainCameron McBrideMark J. McBrideJessica R. McCannRyan McClureDiane McDougaldAndrew McDowellElizabeth A. McGrawK. Kai McKinstrySandra L. McLellanKatherine McMahonMarnix H. MedemaCarlos MedinaPeter MeinickeAlessio MengoniIlhem MessaoudiJulie MeyerKate MichelYusuke MinatoAaron P. MitchellMasatoshi MiyakoshiJennifer M. MobberleyAndrew MoellerJonathan MonkJames J. MoranJacob Moran GiladRobert M. MorrisDouglas MorrisonRafal MostowyVladimir L. MotinPatricia MoweryMaitreyee MukherjeeEmilie E. L. MullerDelia MunteanMarc MussmannJay NadeuNikhil NairYuji NaitoTeru NakatsujiFernando Navarro-GarciaDipti D. NayakJulia NeilsonTiffanie NelsonJeniel E. NettMarkus NettAnthony P. NeumannGraeme William NicolHenrik NilssonAleksandra Nita-LazarJustin R. NodwellTeresa NogueiraJ. Staffan NormarkSpencer V. NyholmPaul Alexander O\u2019BrienMary O\u2019Connell-MotherwayPierre OffreAndrew OgramNiall O\u2019LearyMatthew R. OlmRandall J. OlsenMichelle Ann O\u2019MalleyDana OpulenteAnne-Marie OverstreetEgon Anderson OzerRobert J. PalmerGianni PanagiotouDane ParkerRaghuveer ParthasarathyAlessandro PasseraRob PatroSushmita PatwardhanTalima PearsonPhilip E. PellettBeatriz PenalverHong-Juan PengRita C. PessottiMarie-Agnes PetitDaniel PetrasJennifer Pett-RidgeJody PhelanVanessa PhelanHarold PimentelAzul Pinochet-BarrosConstanze PinskeLuc\u00eda PitaThomas G. PlattPatrick Pl\u00e9siatPrzemyslaw PlocinskiGeorg PohnertArgyris PolitisAlicia Ponte-SucreHannah Beth PooleyPhillip B. PopeSteven L. PorterSambhawa PriyaNan QinWensheng QinZhe-Xue QuanRobert Andrew QuinnKorneel RabaeyMirjana Rajili\u0107-Stojanovi\u0107Thandavarayan RamamurthyKelly RamirezGiordano RampioniMatthew Mark RamseyLutgarde RaskinPhilip N. RatherThomas RatteiGregor ReidOlaya Rendueles GarciaFederico ReyPeter RichardMiles RichardsonLionel Rigottier-GoisMichael Scott RobesonJennifer RoccaJeremy RockKyle H. RohdeCaroline RoperDavid A. RosenAshley A. RossSimon RouxMaxim Rubin BlumLuis A. RubioKelly V. RugglesThomas A. RussoMaurizio RuzziSabrina SaboZakee L. SabreeJon SandersHaley SandersonTodd SandrinAlyson SantoroGuillaume SarrabayrouseBrandon SatinskyJimmy SawLizbeth SayavedraJoy ScariaMark A. SchembriKurt SchesserJoshua P. SchimelAmy K. SchmidAnthony P. SchmittLars SchreiberMatthew O. SchrenkH. Steven SeifertAdnane SellamAditi SenguptaRushina ShahRobert M. Q. ShanksLori R. ShapiroAshok Kumar SharmaXinlei ShengLiat ShenhavFrancesca L. ShortMark W. SilbyJustin D. SilvermanAndrew SimpsonMitchell SingerSteven SingerRohita SinhaTimofey SkvortsovBeate M. SlabyBarth F. SmetsDaniel P. SmithDavid G. E. SmithKim SneppenLindsay SoldenDeguang SongJiangning SongEva C. SonnenscheinUtkarsh SoodJorg SoppaAnja SpangStephen SpiroJoseph E. SprakerJason E. StajichLaura SteindlerGeorge C. StewartFrancesco StratiReed StuddendieckXiaoquan SuDavid SueHikaru SuenagaZheng SunMehul SutharYasuhiro SuzukiWesley D. SwingleyJason B. SylvanIlias TagkopoulosGuylaine TalbotYinjie J. TangGerald William TannockShengce TaoW. Andy TaoJaclyn N. TaroniBradford P. TaylorBen TempertonBenno Herman Ter KuileLuke R. ThompsonYun TianLaura TiptonMehdi ToghiMaya TopfTamas TorokGloria Torres-CortesCarolina TropiniBenjamin John TullySilvia TurroniCarles UbedaBeatrix M. UeberheideSabah Ul-HasanAlex van BelkumPetra Van DammeCarin K. VanderpoolTom Van de WieleWillem van SchaikDouwe van SinderenLuciana Vasquez-PintoTommi VatanenYoshiki Vazquez-BaezaSiddarth VenkateshMarie-Joelle VirolleChris VoigtRobin Voigt-ZuwalaJoseph Thomas WadeIrene Wagner-DoblerAlan W. WalkerMatthew WallensteinGuanghua WangJerry WangJoyce WangWei WangXuya WangYaqiang WangZeneng WangZhang WangDoyle WardWei WeiMargaret WeinrothMartin WelkerNicole WheelerSean P. J. WhelanChristopher WhidbeyMarvin WhiteleyAnisha WijeyesekeraRoland Conrad WilhelmRoli WilhelmAmy D. WillisTomasz WilmanskiJennifer R. WilsonJesse WilsonJeffrey H. WitheyBenjamin WoolstonErik S. WrightRosanna C. T. WrightFuqing WuHao WuMichael WuMin WuYu-Wei WuTodd N. WylieKelly WyresGuoxiang XieJiatao XieChunlan XuPeng XuXuewei XuZhenjiang Zech XuChing-Hong YangFiliz Yarimcam SaglamAlyson L. YeeV. Laxmi R. YeruvaYibing YinYasuo YoshidaJun YuXiaoqian YuZhongtang YuMengting YuanYuanchao ZhanChangyi ZhangFaming ZhangJiachao ZhangKai ZhangLimin ZhangQuan-Guo ZhangWeipeng ZhangZhengguang ZhangZiding ZhangFangqing ZhaoRui ZhaoShijie ZhaoXin-Qing ZhaoDongsheng ZhouWenhao ZhouLiangquan ZhuYan ZhuJun ZouIn 2019, we have seen"} +{"text": "Correction to: Trialshttp://dx.doi.org/10.1186/s13063-019-3304-9Originally published name:Galymgan EleuovCorrected name:Galymzhan YeleuovThe original article has been corrected.Following publication of the original article , the aut"} +{"text": "Healthcare-associated infections and antibiotic resistance\u00a0Clostridium difficile infection in Ireland, 2017Epi-Insight 2019;20(1)January 2019, Irelandhttp://ndsc.newsweaver.ie/epiinsight/1scooyoz373n0ykqgxo0gg?a=1&p=54394726&t=17517774\u00a0Food- and waterborne diseases\u00a0Routine reports of gastrointestinal infections in humans, England and Wales: January and February, 2019Health Protection Report; 13(9)15 March 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/786477/hpr0919_GIs.pdf\u00a0Hepatitides\u00a0Laboratory reports of hepatitis A and C in England and Wales: July to September 2018Health Protection Report; 13(3)25 January 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/773602/hpr0319_hepAC.pdf\u00a0Vaccine-preventable diseases\u00a0Diphtheria in England: 2018Health Protection Report; 13(10)22 March 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/788746/hpr1019_dphthr.pdf\u00a0Tetanus in England: 2018Health Protection Report; 13(10)22 March 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/788744/hpr1019_ttns.pdf\u00a0Pertussis - vaccinating mum is good for baby \u2013 update on antenatal pertussis vaccinationEpi-Insight 2019;20(3)March 2019, Irelandhttp://ndsc.newsweaver.ie/epiinsight/zvpnkwflilyn0ykqgxo0gg?a=1&p=54630557&t=17517774\u00a0The need to achieve high vaccine coverage to sustain rubella elimination in IrelandEpi-Insight 2019;20(3)March 2019, Irelandhttp://ndsc.newsweaver.ie/epiinsight/dgoyhvql9ntn0ykqgxo0gg?a=1&p=54630561&t=17517774\u00a0Laboratory-confirmed cases of measles, rubella and mumps, England: October to December 2018Health Protection Report; 13(8)28 February 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/782191/hpr0819_mmr.pdf\u00a0Laboratory confirmed cases of invasive meningococcal infection (England): October to December 2018Health Protection Report; 13(7)22 February 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/780984/hpr0719_imd.pdf\u00a0Haemophilus influenzae by age group and serotype (England): 2018Laboratory reports of Health Protection Report; 13(7)22 February 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/780976/hpr0719_hmphls-nflnz.pdf\u00a0Pertussis vaccination programme for pregnant women update: vaccine coverage in England, July to September 2018Health Protection Report; 13(7)22 February 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/780991/hpr0719_prntl-prtsss-vc.pdf\u00a0Trends in invasive meningococcal disease in Ireland, 1999-2019Epi-Insight 2019;20(2)February 2019, Irelandhttp://ndsc.newsweaver.ie/epiinsight/tyzbecfnxf6n0ykqgxo0gg?email=true&a=1&p=54506595&t=17517774\u00a0Respiratory diseases\u00a0Outbreak of tuberculosis in a school in Dresden, 2017\u22122018Epidemiologisches Bulletin 11-12/201914 March 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/11_12_19.pdf?__blob=publicationFile\u00a0Legionella pneumophilaProbable case of reinfection with Epidemiologisches Bulletin 1/20193 January 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/01_19.pdf?__blob=publicationFile\u00a0Zoonoses and vector-borne diseases\u00a0Common animal associated infections quarterly report : fourth quarter 2018Health Protection Report; 13(9)15 March 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/786463/hpr0919_zoos.pdf\u00a0Case of chronic Q feverVlaams infectieziektebulletin 2019(1)March 2019, Belgiumhttps://www.zorg-en-gezondheid.be/sites/default/files/atoms/files/VIB%202019-1%20-%20DEF.pdf\u00a0Historical aspects of Q feverVlaams infectieziektebulletin 2019(1)March 2019, Belgiumhttps://www.zorg-en-gezondheid.be/sites/default/files/atoms/files/VIB%202019-1%20-%20DEF.pdf\u00a0Creutzfeldt-Jakob disease (CJD) biannual update (February 2019)Health Protection Report; 13(6)15 February 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/779110/hpr0619_cjd.pdf\u00a0TBE: risk areas in Germany (as of January 2019)Epidemiologisches Bulletin 7/201914 February 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/07_19.pdf?__blob=publicationFile\u00a0Leptospirosis 2017-2018EPI-NEWS 4/5, 20196 February 2019, Denmarkhttps://en.ssi.dk/news/epi-news/2019/no-4-5---2019\u00a0Other\u00a0Pregnancy and infectious diseases in IrelandEpi-Insight 2019;20(3)March 2019, Irelandhttp://ndsc.newsweaver.ie/epiinsight/d95m2dq9kepn0ykqgxo0gg?email=true&a=2&p=54630552&t=17517804\u00a0Group A streptococcal infections: first report of seasonal activity, 2018/19Health Protection Report; 13(8)28 February 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/782182/hpr0819_sf-gas.pdf"} +{"text": "This article has been corrected: The correct author name is given below:Laura Mezquitahttps://doi.org/10.18632/oncotarget.26446Original article: Oncotarget. 2018; 9:37393-37406."} +{"text": "This article has been corrected: In Figure https://doi.org/10.18632/oncotarget.9315Original article: Oncotarget. 2016; 7:51096-51106."} +{"text": "Healthcare-associated infections and antibiotic resistance\u00a0Did we get his message right? A short history of hand hygiene on the occasion of the 200th anniversary of Ignaz Semmelweis\u2019 birthEpidemiologisches Bulletin 18/20183 May 2018, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2018/Ausgaben/18_18.pdf?__blob=publicationFile\u00a0Clostridium difficile outbreak investigationsEpidemiologisches Bulletin 14/20185 April 2018, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2018/Ausgaben/14_18.pdf?__blob=publicationFile\u00a0Food- and waterborne diseases\u00a0Campylobacter infectionsRKI guidance on Epidemiologisches Bulletin 23/20187 June 2018, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2018/Ausgaben/23_18.pdf?__blob=publicationFile\u00a0Shigellosis outbreak at a student accommodation siteInfectieziekten Bulletin 2018; 29(6)June 2018, the Netherlandshttps://magazines.rivm.nl/2018/06/infectieziekten-bulletin/uitbraak-van-shigellose-bij-een-studentenvereniging\u00a0Salmonella and Campylobacter infections 2016-17EPI-NEWS 15/201811 April 2018, Denmarkhttps://www.ssi.dk/English/News/EPI-NEWS/2018/No%2015%20-%202018.aspx\u00a0Giardiasis in a nurseryVlaams Infectieziektebulletin, 4/20172017, Belgiumhttps://www.zorg-en-gezondheid.be/sites/default/files/atoms/files/VIB%202017-4_DEF.pdf\u00a0Hepatitides\u00a0Special edition: National day against viral hepatitisBulletin \u00e9pid\u00e9miologique hebdomadaire, 11/201815 May 2018, Francehttp://invs.santepubliquefrance.fr/beh/2018/11/pdf/2018_11.pdf\u00a0Hepatitis A outbreak in a care homeInfectieziekten Bulletin 2018; 29(4)April 2018, the Netherlandshttps://magazines.rivm.nl/2018/04/infectieziekten-bulletin/hepatitis-uitbraak-op-een-zorgboerderij\u00a0Vaccine-preventable diseases\u00a0Laboratory confirmed cases of measles, mumps and rubella, England: January to March 2018Health Protection Report; 12(19)1 June 2018, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/711941/1918_AA_mmr.pdf\u00a0Vaccine coverage for the GP based catch-up meningococcal ACWY (MenACWY) immunisation programme in England to the end of March 2018Health Protection Report; 12(18)25 May 2018, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/711444/hpr1818_menACWY-vc.pdf\u00a0Preliminary vaccine coverage estimates for the meningococcal B (MenB) immunisation programme for England, update from January to March 2018Health Protection Report; 12(15)27 April 2018, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/703471/hpr1518_menbVC.pdf\u00a0Vaccination rates at school entry examination in Germany, 2016Epidemiologisches Bulletin 19/201819 April 2018, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2018/Ausgaben/16_18.pdf?__blob=publicationFile\u00a0Laboratory-confirmed whooping cough 2017EPI-NEWS 16/201818 April 2018, Denmarkhttps://www.ssi.dk/English/News/EPI-NEWS/2018/No%2016%20-%202018.aspx\u00a0Free measles vaccination for non-immune adults EPI-NEWS 13-14/20184 April 2018, Denmarkhttps://www.ssi.dk/English/News/EPI-NEWS/2018/No%2014%20-%202018.aspx\u00a0Measles in Ireland \u2013 weeks 1-12, 2018Epi-Insight 2018;19(4)April 2018, Irelandhttp://ndsc.newsweaver.ie/epiinsight/1qv34ih6ub810gkzp9yxn5?a=1&p=53187118&t=17517774\u00a0Respiratory diseases\u00a0The 2017/2018 influenza seasonEPI-NEWS 23-24/201813 June 2018, Denmarkhttps://www.ssi.dk/English/News/EPI-NEWS/2018/No%2023-24%20-%202018.aspx\u00a0Spotlight on Human Parvovirus B19Epi-Insight 2018;19(6)June 2018, Irelandhttp://ndsc.newsweaver.ie/epiinsight/tflodsptta910gkzp9yxn5?a=2&p=53465132&t=17517804\u00a0Fatal case in a child related to an influenza outbreak in a kindergartenEpidemiologisches Bulletin 22/201831 May 2018, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2018/Ausgaben/22_18.pdf?__blob=publicationFile\u00a0Sexually transmitted diseases\u00a0Syphilis 2017EPI-NEWS 22, 201830 May 2018, Denmarkhttps://www.ssi.dk/English/News/EPI-NEWS/2018/No%2022%20-%202018.aspx\u00a0Zoonoses and vector-borne diseases\u00a0Special edition: Lyme borreliosis and\u00a0other\u00a0tick-borne diseasesBulletin \u00e9pid\u00e9miologique hebdomadaire, 19-20/201819 June 2018, Francehttp://invs.santepubliquefrance.fr/beh/2018/19-20/pdf/2018_19-20.pdf\u00a0TBE: risk areas in Germany (April 2018)Epidemiologisches Bulletin 17/201826 April 2018, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2018/Ausgaben/17_18.pdf?__blob=publicationFile\u00a0Hantavirus infections in Germany: a review of the 2017 epidemic yearEpidemiologisches Bulletin 15/201812 April 2018, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2018/Ausgaben/15_18.pdf?__blob=publicationFile\u00a0Human case of leptospirosis transmitted from a dogInfectieziekten Bulletin 2018; 29(4)April 2018, the Netherlandshttps://magazines.rivm.nl/2018/04/infectieziekten-bulletin/humane-leptospirose-een-puppy\u00a0Leptospirosis among participants in an obstacle run, Nijlen 2015Vlaams Infectieziektebulletin, 4/20172017, Belgiumhttps://www.zorg-en-gezondheid.be/sites/default/files/atoms/files/VIB%202017-4_DEF.pdf\u00a0Historical aspects of leptospirosisVlaams Infectieziektebulletin, 4/20172017, Belgiumhttps://www.zorg-en-gezondheid.be/sites/default/files/atoms/files/VIB%202017-4_DEF.pdf"} +{"text": "The correct name is: Maria Jose Fuster-RuizdeApodaca.The 11th and August 8th of 2017, showing the following messages: \u201cAre you PrEPared for the pill that prevents HIV? / Help us to improve the health of LGBT community by answering some questions / Are you PrEPared for World Pride?\u201dIn the Design of the questionnaire subsection of the Materials and methods, there is an error in the fourth sentence. The correct sentence is: The survey was advertised through gay-oriented dating apps and the social media of HIV Non-Governmental Organizations (NGO) between June 26In https://www.msssi.gob.es/ciudadanos/enfLesiones/enfTransmisibles/sida/docs/PlanEstrategico2013_2016.pdfReference 2 is spelled incorrectly. The correct reference is: Ministerio de Sanidad, Servicios Sociales e Igualdad. Plan Estrat\u00e9gico de Prevenci\u00f3n y Control de la infecci\u00f3n por el VIH y otras infecciones de transmisi\u00f3n sexual [Internet]. 2013. Available: https://ecdc.europa.eu/sites/portal/files/media/en/publications/Publications/EMIS-2010-european-men-who-have-sex-with-men-survey.pdfThe URL in reference 3 is incorrect. The correct URL is: https://cima.aemps.es/cima/pdfs/ft/04305001/FT_04305001.pdfThe URL in reference 13 is incorrect. The correct URL is: http://www.mscbs.gob.es/ciudadanos/enfLesiones/enfTransmisibles/sida/docs/PROFILAXIS_PREEXPOSICION_VIH.pdfThe URL in reference 17 is incorrect. The correct URL is: Ferrer L, Folch C, Fernandez-Davila P, Garcia A, Morales A, Belda J, et al. was incorrectly included as reference 26. As a result, all subsequent references are misnumbered. References 27\u201329 should be references 26\u201328."} +{"text": "This article has been corrected: Due to errors in image processing, there was an accidental duplication of data in 2250-2262. https://doi.org/10.18632/oncotarget.2948Original article: Oncotarget. 2015; 6:2250\u20132262."} +{"text": "R. Soc. open sci.5, 171529. (Published 27 June 2018). (doi:10.1098/rsos.171529)This correction refers to tables"} +{"text": "Correction to: BMC Bioinformatics (2016) 17:185https://doi.org/10.1186/s12859-016-1013-xFollowing publication of the original article , the autThe links that needs to be updated can be find at the Conclusions section of Abstract and other under Project home page at the bottom.Old link (s):http://metagenomics.atc.tcs.com/compnet/ orhttp://121.241.184.233/compnet/Updated new link:https://web.rniapps.net/compnet/"} +{"text": "This article has been corrected: The corrected author name is given below:Wataru Ichikawa18811-18820. https://doi.org/10.18632/oncotarget.24702Original article: Oncotarget. 2018; 9:"} +{"text": "This article has been corrected: The correct Author name is given below:Eugenia Johnsonhttps://doi.org/10.18632/oncotarget.26047Original article: Oncotarget. 2018; 9:34259-34278."} +{"text": "This article has been corrected: In the supplementary file on page 4, the corrected author name is given below:M. Tischkowitz1127-1133. https://doi.org/10.18632/oncotarget.385Original article: Oncotarget. 2011; 2:"} +{"text": "This article has been corrected: The correct author names are given below:Yueqiang Jiang and Yanling Ma11989-11998. https://doi.org/10.18632/oncotarget.22857Original article: Oncotarget. 2018; 9:"} +{"text": "Nucleic Acids Research, gky768, https://doi.org/10.1093/nar/gky768In Figures"} +{"text": "This article has been corrected: The #2 affiliation information has been updated. The proper institution name is shown below:2Bioinformatics, Miltenyi Biotec GmbH, Bergisch Gladbach, 51429, Germany3183-3197. https://doi.org/10.18632/oncotarget.26900Original article: Oncotarget. 2019; 10:3183\u20133197."} +{"text": "This article has been corrected: During production, the page numbers for this article were listed incorrectly. The page numbers have now been adjusted to show the proper pagination.552-562. https://doi.org/10.18632/oncotarget.190Original article: Oncotarget. 2010; 1:"} +{"text": "Vectors of arboviruses in the state of S\u00e3o Paulo: 30 years of Aedes aegypti and Aedes albopictus\u201d Rev. Saude Publica [online]. 2019, vol.53:84, ISSN 1518-8787, http://dx.doi.org/10.11606/s1518-8787.2019053001264, the RSP corrects the author\u2019s last name.In \u201cWhere you read:Marcia Moreira HolcmamYou should read:Marcia Moreira Holcman Vetores de arboviroses no estado de S\u00e3o Paulo: 30 anos de Aedes aegypti e Aedes albopictus\u201d Rev. Saude Publica [online]. 2019, vol.53:84, ISSN 1518-8787, http://dx.doi.org/10.11606/s1518-8787.2019053001264, a RSP corrige o sobrenome do autor.No artigo. \u201cOnde se lia:Marcia Moreira HolcmamLeia-se:Marcia Moreira Holcman"} +{"text": "This article has been corrected: The corrected author name is given below:Alessandro Sanduzzi19356-19367. https://doi.org/10.18632/oncotarget.25049Original article: Oncotarget. 2018; 9:"} +{"text": "This article has been corrected: In the Materials and Methods section, the data access link is incorrect. In addition, reference 37 was also listed incorrectly. The proper information is given below:http://www.ncbi.nlm.nih.gov/sra/SRP102906).Materials and Methods: The data discussed in this publication have been deposited in the NCBI Sequence Read Archive (SRA) [37] and are accessible through : D19-D21.106948-106961. https://doi.org/10.18632/oncotarget.22157Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: The correct author name is given below:Rosa Blancohttps://doi.org/10.18632/oncotarget.11425Original article: Oncotarget. 2017; 8:29679-29698."} +{"text": "R. Soc. open sci.6, 190418. (Published 10 April 2019). (doi:10.1098/rsos.190418)This correction refers to an error in the abstract. Transcriptome measurements were accidentally changed to \u2018transcriptase measurements\u2019. The correct text appears below.Integrated studies via kinetic metabolic modelling, transcriptome measurements and metabolic profiling were performed on this strain."} +{"text": "This article has been corrected: Due to errors in figure preparation, the data for 13023-13035. https://doi.org/10.18632/oncotarget.24310Original article: Oncotarget. 2018; 9:13029\u201313035."} +{"text": "This article has been corrected: During production, the page numbers for this article were listed incorrectly. The page numbers have now been adjusted to show the proper pagination.563-577. https://doi.org/10.18632/oncotarget.191Original article: Oncotarget. 2010; 1:"} +{"text": "Correction to: BMC Medhttps://doi.org/10.1186/s12916-019-1257-1Author, Chee H. Ng\u2019s middle initial was omitted;The Competing Interests declaration omitted some information regarding author, Chee H. Ng.The original article containeThese two errors have since been corrected."} +{"text": "R. Soc. open sci.4, 170351. (Published 18 October 2017). (doi:10.1098/rsos.170351)SRP041222, PRJNA398316 and PRJNA398198.The third accession number in the data accessibility section of the published paper is incorrect. The correct accession numbers are"} +{"text": "This article has been corrected: The corrected author name is given below:Barbieri Eveline20323-20338. https://doi.org/10.18632/oncotarget.24859Original article: Oncotarget. 2018; 9:"} +{"text": "This article has been corrected: The correct author name is given below:Patricia S.P. Thong19902-19913. https://doi.org/10.18632/oncotarget.15720Original article: Oncotarget. 2017; 8:"} +{"text": "TETFund (Tertiary Education Trust Fund): Institution based TETFund grant. [TETFUND/DESS/UNI/NSUKKA/2017/RP/VOL.I].The original article mistaken"} +{"text": "This article has been corrected: During the assembly of the Figure https://doi.org/10.18632/oncotarget.24815Original article: Oncotarget. 2018; 9:17631-17644."} +{"text": "Amblyomma mixtum ticks were misidentified as A. sculptum in Rickettsia africae and Novel Rickettsial Strain in Amblyomma spp. Ticks, Nicaragua, 2013 . The article has been corrected online (https://wwwnc.cdc.gov/eid/article/24/2/16-1901_article)."} +{"text": "The Pan African Medical Journal. 2014;19:50. doi:10.11604/pamj.2014.19.50.4683.This corrigendum corrects article The author affiliation in the published article \u201cService"} +{"text": "The co-first author\u2019s name is spelled incorrectly. The publisher apologizes for the error. The correct name is: Julia TCW. The correct citation is:https://doi.org/10.1371/journal.pone.0213374.Bowles KR, TCW J, Qian L, Jadow BM, Goate AM (2019) Reduced variability of neural progenitor cells and improved purity of neuronal cultures using magnetic activated cell sorting. PLoS ONE 14(3): e0213374."} +{"text": "This article has been corrected: The correct figure https://doi.org/10.18632/oncotarget.3218Original article: Oncotarget. 2015; 6:9820-9833."} +{"text": "This article has been corrected: The corrected author name is given below:Lei Shi26711-26723. https://doi.org/10.18632/oncotarget.25475Original article: Oncotarget. 2018; 9:"} +{"text": "R. Soc. open sci.5, 171430. (Published online 14 March 2018). (doi:10.1098/rsos.171430)In the published paper, the last author's name is presented incorrectly. The last author's name is Mahmood M. S. Abdullah."} +{"text": "That would not have been possible had it not been for the scientists who have dedicated their time to supporting the fair and unbiased review of the science presented in mSystems. We are indebted to the many people who have contributed to article reviews over the last 3 years. What you all have sacrificed in time has been repaid in furthering the pursuit of knowledge in a format that is freely accessible to the whole world.Stephen T. AbedonJonathan Miles AdamsMark D. AdamsSarah AdesPamela R. F. AdkinsEvelien M. AdriaenssensVinayak AgarwalBrian M. M. AhmerAlexander AksenovMd Tauqeer AlamMads AlbertsenPietro AlifanoRobert AllakeRaul A. AlmeidaEric AltermannKatherine R. AmatoAnthony AmendAmnon AmirKarthik AnantharamanJamila Anba-MondoloniCheryl P. AndamHeidi M. AndersenRika AndersonRaul AndinoLargus T. AngenentKym S. AntonationAyako Aoki-YoshidaCristian ApetreiMichael A. ApicellaHector Arguello-RodriguezMasanori AritaJean ArmengaudMagnus \u00d8. ArntzenMario Arrieta-OrtizGraeme Trevor AttwoodJennifer AuchtungJulian Avila PachecoMartin Iain BahlAbhay BajajBrett J. BakerMatthew G. BakkerChandra Shekhar BakshiDavid A. BaltrusCarlos BarajasBridget BarkerStephen BarnesFrancisco Barona-GomezJeremy J. BarrJeffrey E. BarrickAmanda BarryDouglas H. BartlettAnke BeckerSylvia Becker-DrepsEric Daniel BecraftPeter Trip BeerninkMarcel A. BehrGordon Morse BennettIsabelle BenoitGabriele BergIvan BergHans C. BernsteinClaire BertelliStefan BertilssonErin BertrandWolfgang BeyerShantanu BhattKyle BibbyElisabeth M. BikSteven BillerFabrice Vincent BiotJohannes Bj\u00f6rkJohanna Bj\u00f6rkrothJill R. BlankenshipSteve BlazewiczRan BlekhmanRobert M. BlumenthalGregory BokinskyNicholas Andrew BokulichTobias BollenbachGregory BonitoRichard BonneauErika BonsagliaRafaella C. Bonugli-SantosAnna BothBrian BothnerDavid G. BourneJohn P. BowmanSean F. BradyAxel A. BrakhageJonathan BraunAsker Daniel BrejnrodOrianna BretschgerTess Elizabeth BrewerLazzaro BrianDavid S. BriskeyBrandon BrooksPaul BrooksC. Titus BrownDaniel R. BrownJeremy C. BrownlieJames E. BruceDonald A. BryantSamuel Joseph BrysonNicole R. BuanAlison BuchanMichael J. BuchmeierCarmen BuchrieserSara Ashley BumgardnerAdam R. BurnsBrendan Paul BurnsFrederic D. BushmanKyle C. CadyBen CallahanBenjamin J. CallahanStephen J. CallisterAndres Mauricio Caraballo RodriguezValerie Jean CarabettaPaul CariniErin CarlsonRoss P. CarlsonGilda CarvalhoMagali Chab\u00e9Woo-Suk ChangMichael S. ChausseeJun ChenTingtao ChenLei ChengLeonid ChindelevitchPeter ChiversSidharth ChopraJoseph Alexander Christie-OlezaGeremy C. ClairTom ClavelLuca CocolinDavid A. CoilAndre M. ComeauJohn CommonLaurie E. ComstockJonathan ConwayTyrrell ConwayVaughn S. CooperOtto X. CorderoElizabeth K. CostelloNatacha CoutoDon A. CowanMichael CoxKatharine CoyteJohn CryanCatalina Cuellar-GempelerJulia CuiChristina CuomoDennis G. CvitkovitchChristiane DahlColin DaleRemus DameJeffrey L. DanglF. DarfeuilleJulian E. DaviesJames DavisStacy L. DeblasioVal\u00e9rie De Cr\u00e9cy-LagardMichael DeemCarlotta De FilippoNicholas R. De LayCeline Delbes-PausEdward F. DelongYe DengMuriel DerrienPetra DerschMahesh DesaiAdam M. DeutschbauerKen DewarGregory J. DickChristian DienerDirk P. DittmerJeremy A. DodsworthClaudio DonatiYiran DongMohamed S. DoniaIan DonohueAngela E. DouglasGavin M. DouglasJaquelin P. DudleyTim J. DumonceauxJohn DunbarKatherine R. DuncanTaylor DunivinAshlee M. EarlBrian Joseph EddieLaura EmeBruce D. EricksonCassandra Lane EttingerAmandine EverardVanessa EzenwaAndrew James FabichLiam J. FanningKaroline FaustGabriel da Rocha FernandesLucia FernandezNoah FiererOmri M. FinkelNahuel FittipaldiErik K. FlemingtonHarry J. FlintLucile M. Floeter-WinterYuriy FofanovEdan FoleyMarco FondiJoseph Che ForbiAdriana ForeroKristoffer ForslundSamuel Charles ForsterKonrad Ulrich F\u00f6rstnerPaul ForsytheJamie S. FosterZachary FosterFiona FouhyMiguel FradaPedro FradePilar FrancinoEelco FranzSebastian FrauneStephen J. FreeKatie FreelPengcheng FuJed FuhrmanTamas GaalMihaela GadjevaFrancesca GaggiaMarco GalardiniJinshan GaoJos\u00e9 Luis Garc\u00edaJeffrey G. GardnerDaniel GarridoCaroline Attardo GencoC. R. Robert GeorgeLaurel J. GershwinAndrew GewirtzRaad GharaibehPaolo GhensiTravis GibsonLaura GlendinningGregory B. GloorJon D. GoguenShan GohSusan S. GoldenMaria Adelaida GomezDavid J. GonzalezAndrew GoodmanUri GophnaStephen V. GordonDavid C. GraingerJeffrey A. GralnickLeigh GreathouseStefan J. GreenJulian GriffinChristopher A. GulvikMichelle Gwinn-GiglioElaine M. HaaseMarkus HaberMurray HackettMartin W. HahnMehrdad HajibabaeiLindsay J. HallNafiz HamidRobert E. W. HancockKim M. HandleyMatthew J. HarkeJoe HarrisonBernhard HauboldKeith HazeltonIan HeadBrian P. HedlundMatthias HeinemannMichael HensonUte HentschelMalte HeroldAnat A. HerskovitsRobert L. HettichThomas HitchWilliam E. HolbenGeorgina Louise HoldJohannes HolertEmily B. HollisterMatthias HornJianzhong HuSarah K. HuShi HuangJudith M. H\u00fcbschenLaura A. HugEili HuhtamoBruce A. HungateDana E. HuntC. Neil HunterDaniel HusonEmbriette R. HydeTadayuki IwaseJacques IzardJonathan L. JacobsCrystal JaingMichael JanechIan B. JefferyPoul Erik JensenPatricio JeraldoDing Jun JinSeth G. JohnKay O. JohswichIan JointKathryn M. JonesKirsten JungNadeem O. KaakoushKrishna (Rani) KalariVipin Chandra KaliaMarina G. KalyuzhnayaTaro KamagakiDae Wook KangWei-Chun KaoWojciech KarlowskiPurna Chandra KashyapAnne-Kristin KasterElena KazamiaSean KearneyScott T. KelleyColleen KelloggScott KenneySaeed KhanKhashayarsha KhazaieMary L. KillianMinsuk KimPan-Jun KimBenjamin C. KirkupTodd KittenJonathan KlassenSilke KleeVanja Klepac-CerajDan KnightsKevin KohlKonstantinos T. KonstantinidisEugene V. KooninOmry KorenNicole Marie KoropatkinDavid KoslickiAleksandar KosticPetia KovatchevaJens H. KuhnPrashant KumarRoshan KumarDeepak KumaresanElizabeth M. KutterBrandon LaBumbardDouglas J. LaCountIsabelle Laforest-LapointeLeo LahtiMichael LammersAmy LaneJenna M. LangPascal LapierreAdi LavyVladimir LazarevicSarah LebeerYoann LeBretonFredrick J. LeeKyongbum LeePhilippe Claude LefebvreJose LemosVanessa LeoneSabine LeroyHuiying LiTami LiebermanGipsi Lima MendezKai-Shu LingPushpinder LittNan LiuRichard LongneckerCedric LoodStilianos LoucaPetra LouisClaude LoverdoKun LuMichael LynchBoris MacekBarbara J. MacGregorJohn MacSharryJuliette MadanStefania MagnusdottirFinlay MaguireFr\u00e9d\u00e9ric Mah\u00e9Vladimir MakarenkovThulani P. MakhalanyaneMeenakshi MalikElizabeth MallottOhad ManorCostas D. MaranasMaria L. MarcoAngela MarcobalJeffrey MarlowNatalie MarshallRebeca MartinAdam MartinyBernd MasepohlAndrea MasottiJessica R. McCannRyan McClureBeth McCormickJohn McCutcheonJason E. McDermottDaniel McDonaldAndrew McDowellMichael J. McInerneyLuke McKayJeffrey Scott McLeanKatherine McMahonAlan McNallyPatrick J. McNamaraJames F. MeadowMarnix H. MedemaMonica MedinaConor MeehanJay MelliesAlexey V. MelnikBodo MelnikAkram MendezAlessio MengoniShiri MeshnerIlhem MessaoudiJessica MetcalfThomas MetzKim MilferstedtAndrew D. MillardShujiro MinamiTim MiyashiroJennifer M. MobberleyAndrew MoellerDebasisa MohantyHosein MohimaniJonathan MonkChristophe MonnetLisa MooreNathaniel John MoormanCharles P. MoranMary Ann MoranMaria Aparecida Scatamburlo MoreiraXochitl C. MorganJenna Morgan-LangAndrey MorgunCindy E. MorrisJ. Jeffrey MorrisRobert M. MorrisElke M\u00fchlbergerMaitreyee MukherjeeMartha H. MulksAlvaro MunozVivek MutalikHiroaki NakaGerard J. NauLama NazzalJulia NeilsonWilliam C. NelsonCamilla Lothe NesboMarkus NettTerry Fei Fan NgNancy N. NicholsJens NielsenJeppe Lund NielsenOlivia D. NigroHiroshi NikaidoAleksandra Nita-LazarKrishna NiyogiCecilia NoeckerKenneth M. NollTorbj\u00f6rn Nor\u00e9nHoward OchmanMary O'Connell-MotherwayTadgh O'CroininMarco Rinaldo OggioniStephen G. OliverHuub J. M. Op Den CampAharon OrenHiroyuki OshiumiElizabeth A. OttesenAbdullah OzerOlga OzolineKeith PaarpornOleg PaliyBrent PalmerRobert J. Palmer, Jr.Chongle PanLaura ParfreyJohn ParkinsonKiran R. PatilVaishnavi PattabiramanAndrew D. PattersonJoseph N. PaulsonChristopher PeacockShyamal D. PeddadaBeatriz PenalverJohn PendersBingyin PengCharles Pepe-RanneyMatthew PerisinJakob PernthalerHannes PeterJason PetersStefan PfeifferVanessa PhelanJo PhilipsCaroline C. PhilpottPascal PineauAmeet J. PintoJames M. PipasGerman PlataAngela C. PoolePhillip B. PopeAlexa A. PragmanOm PrakashMichael B. PrenticeLance B. PriceMorgan N. PriceNathan PriceSladjana PrisicCatherine PutontiMeng QiJianming QiuYuanyuan QuZhe-Xue QuanRobert Andrew QuinnElisabeth A. RaleighSrinivasan RamachandranMatthew Mark RamseyChristopher RaoPhilip N. RatherFrederic RaymondGregor ReidPierre RenaultFrancis RepoilaScott RiceJason M. RidlonLionel Rigottier-GoisDaina L. RingusJudith RisseAdam RiversMichael Scott RobesonDmitry A. RodionovGeraint B. RogersDavid A. RosenIda RosenkrandsJohannes RouskSimon RouxTaniya Roy ChowdhuryBen RubinSteven RutherfordRatul SaikiaNina R. SalamaHassan SameeAlvaro SanchezJon Sanders\u00c1lvaro San MillanBeng San YeohPauline Deirdre ScanlanPatrick D. SchlossThomas Mitchell SchmidtLynn SchrimlThomas SchwederHerbert P. SchweizerHenning SeedorfAnna Maria SeekatzAlecia SepterReed S. ShabmanWilliam M. ShaferRushina ShahJason W. ShapiroOm Prakash SharmaLaura Jayne SherrardRafael Silva-RochaJustin D. SilvermanPallavi SinghYogendra SinghRohita SinhaLindsey M. SoldenAbraham L. SonensheinSe Jin SongJustin SonnenburgJ\u00f6rg SoppaDiana SousaVanessa SperandioDaan R. SpethJoseph Evan SprakerStefan SpringEric V. StabbPierre StallforthLisa Y. SteinRalf SteuerChris StewartO. Colin StineUlrich StinglDaniel StoebelXiaoquan SuGarret SuenMatthew SullivanFengzhu SunJun SunMing SunShinichi SunagawaMichael G. SuretteMichiko E. TagaIlias TagkopoulosGuylaine TalbotR\u00e9gine TalonHideyuki TamakiMing TanPatrick TangGerald William TannockSuvi TaponenMike TaylorRobert ThackerCasey TheriotNicholas R. ThomsonTamas TorokSusannah Green TringePankaj TrivediTommy Tsan-Yuk LamTamir TullerBenjamin John TullyJenny TungKieran TuohlyHanne L. P. TytgatMario UchimiyaHidetoshi UrakawaTadasu UrashimaSergio UzzauJuan Jos\u00e9 Valdez-Alarc\u00f3nJairam K. P. VanamalaLia van der HoekJustin van der HooftDaniel van der LelieDavid van DuinGeertje van KeulenTricia A. Van LaarGary VanzinPatricia Sampaio Tavares VerasKevin VerstrepenEmily VogtmannWilliam G. WadeIrene Wagner-DoblerMatthew K. WaldorLevi WaldronAlan W. WalkerMatthew WallensteinWilliam Anton WaltersChao-Min WangGuanghua WangPauline W. WangChristopher WardDoyle WardAlex D. WashburneKenneth WasmundMick WatsonGrzegorz WegrzynPamela WeisenhornHartmut WekerleGeorge F. WellsMichiel WelsJeffrey WernerLaura WeyrichWilliam B. WhitmanShannon WhitmerGiovanni WidmerSivaramesh WigneshwerarajAnnegret WildeMichael James WilkinsBenjamin P. WillingAmy WillisJennifer R. WilsonKathryn WingleeBenjamin E. WolfeBen J. WoodcroftJeffrey A. WoodsColin WorbyAaron T. WrightErik S. WrightGerard D. WrightKelly WrightonHao WuMichael WuJoao XavierZhenjiang Zech XuLuying XunJane Y. YangStephanie A. YarwoodAlyson YeePelin YilmazYanbin YinSang Sun YoonMengting YuanJoseph P. ZackularElena ZaikovaJin ZengKarsten ZenglerKai ZhangLixin ZhangMeiling ZhangWenjun ZhangHaitao ZhaoLiang ZhaoYingming ZhaoHuaijun ZhouJizhong ZhouLuchang ZhuErik R. ZinserGang ZouPeter ZuberWe are coming to the end of 2018, which has seen"} +{"text": "This article has been corrected: The author names below were unintentionally reversed in the listing. The correct author names are as follows:Alshaimaa AbdelmoezVeronika Maria MetzlerDaniel DejacoHerbert RiechelmannJozef DudasIra-Ida Skvortsova3641-3652. https://doi.org/10.18632/oncotarget.23248Original article: Oncotarget. 2018; 9:3641\u20133652."} +{"text": "This article has been corrected: The corrected author name is given below:Youping Deng13407-13422. https://doi.org/10.18632/oncotarget.24388Original article: Oncotarget. 2018; 9:"} +{"text": "There are errors in the Author Contributions. The correct contributions are: Conceptualization: OF MA GM DA. Data curation: OF MA DA AO. Formal analysis: OF MA GM. Investigation: OF MA GM. Methodology: OF MA. Software: OF MA. Supervision: MA GM. Writing\u2014original draft: OF MA GM DA AO. Writing\u2014review & editing: OF MA GM DA.There are errors in the Funding statement. The correct Funding statement is as follows: The authors received no specific funding for this work.S1 File(DOCX)Click here for additional data file."} +{"text": "This article has been corrected: During the assembly of Figure 164-178. https://doi.org/10.18632/oncotarget.10516Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: Figures 7489-7501. https://doi.org/10.18632/oncotarget.9841Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: The correct author name is given below:E. Kate Kemsleyhttps://doi.org/10.18632/oncotarget.26022Original article: Oncotarget. 2018; 9:33278-33289."} +{"text": "This article has been corrected: The new affiliation no.14 is given below:Chiang-Wen Lee1414Department of Rehabilitation, Chang Gung Memorial Hospital, Chia-Yi 61363, Taiwan28342-28358.https://doi.org/10.18632/oncotarget.16058Original article: Oncotarget. 2017; 8:28342\u201328358."} +{"text": "Phil. Trans. R. Soc. A376, 20170449. (Published online 29 October 2018). (doi:10.1098/rsta.2017.0449)In figure 6, there is an error with one of the waveforms. The correct version of figure 6 is below."} +{"text": "J Radiat Res 2019; 60:159\u2013160. https://doi.org/10.1093/jrr/rry082.Corrigendum to: Tsukasaki A, Taira Y, Orita M, et al. Seven years post-Fukushima: long-term measurement of exposure doses in Tomioka Town. The author name Akira Tsukazaki was incorrect. It should have been Akira Tsukasaki. This has been corrected online."} +{"text": "This article has been corrected: The correct name of the 4th author is as follows:Lisbet Rosenkrantz H\u00f6lmich27062-27074. https://doi.org/10.18632/oncotarget.16003Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: Authors Anil K. Rustgi and Erica L. Carpenter contributed equally as corresponding authors. The footnote to the author list indicating both individuals as co-corresponding authors was unintentionally omitted. The proper footnote has now been added as shown below:** Co-corresponding authors3592-3604.https://doi.org/10.18632/oncotarget.26911Original article: Oncotarget. 2019; 10:3592\u20133604."} +{"text": "R. Soc. open sci.5, 181457. (Published 1 November 2018). (doi:10.1098/rsos.181457)b) was missing. The corrected figure and caption appear below.This correction refers to an error in figure 6. The key to symbols for ("} +{"text": "This article has been corrected: Due to errors in image assembly, the flow cytometry profiles depicting the isotype matched control (IMC) for CD49f on both scramble (scr) and miR-100 reported in 2315-2330 . https://doi.org/10.18632/oncotarget.2962Original article: Oncotarget. 2015; 6:2315\u20132330."} +{"text": "The Pan African Medical Journal. 2015;20:437. doi:10.11604/pamj.2015.20.437.5660.Ce Corrigendum modifie l\u2019article original La version originale de cet article contientLes noms des auteurs: Gbekley Efui Holaly change pour Holaly Gbekley Efui. Karou Damintoti Simplice change pour Damintoti Simplice Karou. Gnoula Charlemagne change pour Charlemagne Gnoula. Agbodeka Kodjovi change pour Kodjovi Agbodeka. Anani Kokou change pour Kokou Anani. Tchacondo Tchadjobo change pour Tchadjobo Tchacondo. Agbonon Amegnona change pour Amegnona Agbonon. Batawila Komlan change pour Komlan Batawila. Simpore Jacques change pour Jacques Simpore."} +{"text": "Xanthomonas genus includes many Gram-negative plant-associated bacteria. Here, we report a virulent Xanthomonas siphophage called Samson. A siphophage isolated from sewage, Samson contains a 43,314-bp genome with 58 predicted genes. Samson has high nucleotide identity with Pseudomonas phage PaMx42.The Xanthomonas genus includes many Gram-negative plant-associated bacteria. Here, we report a virulent Xanthomonas siphophage called Samson. A siphophage isolated from sewage, Samson contains a 43,314-bp genome with 58 predicted genes. Samson has high nucleotide identity with Pseudomonas phage PaMx42.The Xanthomonadaceae are a diverse family of plant-associated Gram-negative bacteria , as described by Ahern et al. (https://www.bioinformatics.babraham.ac.uk/projects/fastqc/). After trimming with the FastX Toolkit v0.0.14 (http://hannonlab.cshl.edu/fastx_toolkit/), the phage genome was assembled into a single contig with SPAdes v3.5.0 (using default parameters) at a coverage of 182.7-fold wastewater samples collected in College Station, Texas. Samson was propagated by the soft-agar overlay method of Adams on a ricn et al. . Morpholn et al. . Genomicn et al. . An Illu2.7-fold \u201311. Puta.7-fold \u2013. Gene fuATCC PTA-101, as d7-fold \u2013\u2013. Potenti7-fold \u2013\u2013. Structu7-fold \u2013\u2013. The gen7-fold \u2013\u2013. All tooaxy-pub/ , 21.Pseudomonas phage PaMx42 (GenBank accession no. JQ067092), with which it shares 56 similar proteins.Samson is a 43,314-bp siphophage with a G+C content of 54.47%. The 94.94% coding density derives from 58 predicted protein-coding genes. PhageTerm predicts that Samson uses a headful-type packaging mechanism. The most closely related phage to Samson, with 95.9% nucleotide identity, is MN062187, BioProject no. PRJNA222858, SRA no. SRR8892199, and BioSample no. SAMN11411460.The genome sequence and associated data for phage Samson were deposited under GenBank accession no."} +{"text": "This article has been corrected: While analyzing the same patient cohort for different markers, the authors realized a mistake in 69976-69990. https://doi.org/10.18632/oncotarget.12099Original article: Oncotarget. 2016; 7:69976\u201369990."} +{"text": "Chelydra serpentina) and alligator (Macrochelys temminckii) snapping turtles. PLoS ONE 14(6): e0217626. https://doi.org/10.1371/journal.pone.0217626.The third author\u2019s name is spelled incorrectly. The correct name is: Lancia Darville. The correct citation is: Baker S, Kessler E, Darville L, Merchant M (2019) Different mechanisms of serum complement activation in the plasma of common ("} +{"text": "Healthcare-associated infections and antibiotic resistance\u00a0Mycobacterium chimaera infection associated with the use of heater-cooler equipment is a novel and rare, but important differential diagnosisProlonged non-specific disease in cardiac valve-operated people: EPI-NEWS 40, 201925 October 2019, Denmarkhttps://en.ssi.dk/news/epi-news/2019/no-40---2019\u00a0Frequency, characteristics and distribution of MRSA in Germany: situation for 2017\u20132018Epidemiologisches Bulletin 42/201917 October 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/42_19.pdf?__blob=publicationFile\u00a0Suspected and laboratory confirmed reported norovirus outbreaks in hospitals (England) and norovirus laboratory reports : weeks 36 to 39, 2019Health Protection Report; 13(36)11 October 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/838807/hpr3619_noro.pdf\u00a0Klebsiella pneumoniae with OXA-48 and NDM-1 carbapenemases and colistin resistance in Mecklenburg-VorpommernInformation about the occurrence of Epidemiologisches Bulletin 40/20192 October 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/40_19.pdf?__blob=publicationFile\u00a0Food- and waterborne diseases\u00a0Listeria monocytogenes outbreak resolvedProlonged local EPI-NEWS 45, 201920 November 2019, Denmarkhttps://en.ssi.dk/news/epi-news/2019/no-45---2019\u00a0Routine reports of gastrointestinal infections in humans, England and Wales: September and October 2019Health Protection Report; 13(40)8 November 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/845643/hpr4019_GIs.pdf\u00a0Listeria monocytogenes sequence cluster type 2521 (Sigma1) in GermanyListeriosis outbreak with Epidemiologisches Bulletin 41/201910 October 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/41_19.pdf?__blob=publicationFile\u00a0Increase in typhoid notifications in travellers returning from PakistanEpi-Insight 2019;20(10)October 2019, Irelandhttp://ndsc.newsweaver.ie/epiinsight/1npbbpsg5wm?a=1&p=55721182&t=17517774\u00a0Hepatitides\u00a0Acute hepatitis B: national enhanced surveillance report January to June 2019Health Protection Report; 13(38)25 October 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/841889/hpr3819_ct-hepB.pdf\u00a0Special edition: National day against viral hepatitis, 2019Bulletin \u00e9pid\u00e9miologique hebdomadaire, 24-25/201924 September2019, Francehttp://beh.santepubliquefrance.fr/beh/2019/24-25/index.html\u00a0Vaccine-preventable diseases\u00a0Pertussis vaccination programme for pregnant women update: vaccine coverage in England, April to September 2019Health Protection Report; 13(41)2 December 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/849023/hpr4119_AA_prntl-prtsss-vc.pdf\u00a0Invasive meningococcal disease in England: annual laboratory confirmed reports for epidemiological year 2018 to 2019Health Protection Report; 13(38)29 October 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/842368/hpr3819_IMD-ann.pdf\u00a0Meningococcal disease 2018EPI-NEWS 40, 201925 October 2019, Denmarkhttps://en.ssi.dk/news/epi-news/2019/no-40---2019\u00a0Nationwide whooping cough epidemicEPI-NEWS 38, 20198 October 2019, Denmarkhttps://en.ssi.dk/news/epi-news/2019/no-38---2019\u00a0Laboratory confirmed cases of pertussis (England): April to June 2019Health Protection Report; 13(34)30 September 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/835104/hpr3419_prtsss.pdf\u00a0Investigation of the Bavarian State Office for Health and Food Safety on tuberculosis among asylum seekers in BavariaEpidemiologisches Bulletin 39/201926 September 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/39_19.pdf?__blob=publicationFile\u00a0Respiratory diseases\u00a0Legionellosis in the spotlightEpi-Insight 2019;20(11)November 2019, Irelandhttp://ndsc.newsweaver.ie/epiinsight/16vaazyleppimcmkeer4wk?a=1&p=55896984&t=17517774\u00a0Focused issue: InfluenzaBulletin \u00e9pid\u00e9miologique hebdomadaire, 28/201921 October 2019, Francehttp://beh.santepubliquefrance.fr/beh/2019/28/index.html\u00a0Sexually transmitted diseases\u00a0HIV studies and projects at the Robert Koch InstituteEpidemiologisches Bulletin 49/20195 December 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/49_19.pdf?__blob=publicationFile\u00a0Special edition: Epidemiological situation and screening for HIV and other STIsBulletin \u00e9pid\u00e9miologique hebdomadaire, 31-32/201926 November 2019, Francehttp://beh.santepubliquefrance.fr/beh/2019/31-32/index.html\u00a0HIV 2018EPI-NEWS 44, 201920 November 2019, Denmarkhttps://en.ssi.dk/news/epi-news/2019/no-44---2019\u00a0Estimated number of new HIV infections and the total number people living with HIV in Germany, status at the end of 2018Epidemiologisches Bulletin 46/201914 November 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/46_19.pdf?__blob=publicationFile\u00a0Monitoring community HIV testing in IrelandEpi-Insight 2019;20(11)November 2019, Irelandhttp://ndsc.newsweaver.ie/epiinsight/12tu3c2wui1imcmkeer4wk?a=1&p=55896983&t=17517774\u00a0Chlamydia 2018EPI-NEWS 38, 20198 October 2019, Denmarkhttps://en.ssi.dk/news/epi-news/2019/no-38---2019\u00a0Zoonoses and vector-borne diseases\u00a0New autochthonous case of West Nile virus infection has been confirmedEpidemiologisches Bulletin 44/201931 October 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/44_19.pdf?__blob=publicationFileFirst case of West Nile virus infection transmitted by mosquitoes in GermanyEpidemiologisches Bulletin 40/20192 October 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/40_19.pdf?__blob=publicationFile\u00a0Other\u00a0Travel-associated diseases 2018Epidemiologisches Bulletin 48/201928 November 2019, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2019/Ausgaben/48_19.pdf?__blob=publicationFile\u00a0Laboratory surveillance of pyogenic and non-pyogenic streptococcal bacteraemia in England, Wales and Northern Ireland: 2018Health Protection Report; 13(41)20 November 2019, United Kingdomhttps://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/847147/hpr4119_pnp-strptccc.pdf\u00a0Occurrence of enterovirus D68EPI-NEWS 39, 201910 October 2019, Denmarkhttps://en.ssi.dk/news/epi-news/2019/no-39---2019"} +{"text": "This article has been corrected: The correct 10 th reference is given below:https:doi.org/10.1002/cncr.31791.10. Eyre TA, et al. Cancer. 2019; 125:99-108. https://doi.org/10.18632/oncotarget.26460Oncotarget. 2018; 9:37280-37281."} +{"text": "This article has been corrected: The correct author name is given below:Shaojin Zhanghttps://doi.org/10.18632/oncotarget.15841Original article: Oncotarget. 2017; 8:27892-27903."} +{"text": "This article has been corrected: During the assembly of 69351-69361. https://doi.org/10.18632/oncotarget.14540Original article: Oncotarget. 2017; 8:69351\u201369361."} +{"text": "J. R. Soc. Interface15, 20170871 (Published online 16 May 2018) (doi:10.1098/rsif.2017.0871)The funding statement and reference [10] should be revised as follows:"} +{"text": "There are errors in the Author Contributions. The correct contributions are: Conceptualization: AMA MC. Formal analysis: MSM AMA JAH MC LB AGL. Funding acquisition: MC AMA. Investigation: MSM JAD AE MTB LJF AMA MC. Methodology: MSM AMA JAH MC. Project administration: MC AMA. Supervision: AMA MC. Writing \u2013 original draft: MSM AMA. Writing \u2013 review and editing: MSM AMA MC."} +{"text": "R. Soc. open sci.5 171029. (Published Online 10 January 2018) (doi:10.1098/rsos.171029)https://doi.org/10.5061/dryad.t2ch6.2.The Dryad link given in the data accessibility statement of this article is incorrect. The correct link is as follows:"} +{"text": "Correction to: Genome Biol (2005) 6:R102https://doi.org/10.1186/gb-2005-6-12-r102Following publication of the original article , the folThe actin control panel in Fig.\u00a03 of this paper is reproduced from Fig.\u00a07 of Tour\u00e9 et al., 2004 [This correction article also provides alternate correspondence email addresses: aminata.toure@inserm.fr; P.J.I.Ellis@kent.ac.uk"} +{"text": "This article has been corrected: In 8635-8647. https://doi.org/10.18632/oncotarget.3249Original article: Oncotarget. 2015; 6:8635\u20138647."} +{"text": "This article has been corrected: The corrected author name is given below:Sravya Gourishetti11803-11816. https://doi.org/10.18632/oncotarget.7730Original article: Oncotarget. 2016; 7:"} +{"text": "This article has been corrected: The correct author information is given below:Amanda M. Saratsis37112-37124. https://doi.org/10.18632/oncotarget.26430Original article: Oncotarget. 2018; 9:"} +{"text": "This article has been corrected: The image for #4-3 in Figure https://doi.org/10.18632/oncotarget.16598Original article: Oncotarget. 2017; 8:39087-39100."} +{"text": "This article has been corrected: The correct Author name is given below:Vicky L. Fretwell27104-27116. https://doi.org/10.18632/oncotarget.25497Original article: Oncotarget. 2018; 9:"} +{"text": "This article has been corrected: The last name of the 4th author was spelled incorrectly. The proper spelling is given below:1Heather Lillemoe584-594. https://doi.org/10.18632/oncotarget.26549Original article: Oncotarget. 2019; 10:584\u2013594."} +{"text": "This article has been corrected: Due to a production error, the figures in this paper were mislabeled. The correct figure legends are given below:https://doi.org/10.18632/oncotarget.17634Original article: Oncotarget. 2017; 8:58709-58727."} +{"text": "The Scientific World Journal has retracted the article titled \u201cTargeting Mitochondria as Therapeutic Strategy for Metabolic Disorders\u201d [sorders\u201d . The arthttps://www.liebertpub.com/doi/full/10.1089/ars.2009.2531 [\u201cMitochondrial Dysfunction in Diabetes: From Molecular Mechanisms to Functional Significance and Therapeutic Opportunities,\u201d by William I. Sivitz and Mark A. Yorek published in Antioxidants & Redox Signaling, vol. 12, no. 4, 537\u2013577 pages, DOI: 10.1089/ars.2009.2531,"} +{"text": "This article has been corrected: The correct author name is given below:Mariano Bizzarrihttps://doi.org/10.18632/oncotarget.25867Original article: Oncotarget. 2018; 9:31842-31860."} +{"text": "Chlamydia trachomatis PgP3 antibody correlates with time since infection and number of previous infections. PLoS ONE 13(12): e0208652. https://doi.org/10.1371/journal.pone.0208652The second author\u2019s name is spelled incorrectly. The correct name is: Stephanie J. Migchelsen. The correct citation is: Blomquist PB, Migchelsen SJ, Wills G, McClure E, Ades AE, Kounali D, et al. (2018) Sera selected from national STI surveillance system shows"} +{"text": "Although the Concise Guide represents approximately 400 pages, the material presented is substantially reduced compared to information and links presented on the website. It provides a permanent, citable, point\u2010in\u2010time record that will survive database updates. The full contents of this section can be found at http://onlinelibrary.wiley.com/doi/10.1111/bph.14750. Nuclear hormone receptors are one of the six major pharmacological targets into which the Guide is divided, with the others being: G protein\u2010coupled receptors, catalytic receptors, enzymes and transporters. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading. The landscape format of the Concise Guide is designed to facilitate comparison of related targets from material contemporary to mid\u20102019, and supersedes data presented in the 2017/18, 2015/16 and 2013/14 Concise Guides and previous Guides to Receptors and Channels. It is produced in close conjunction with the International Union of Basic and Clinical Pharmacology Committee on Receptor Nomenclature and Drug Classification (NC\u2010IUPHAR), therefore, providing official IUPHAR classification and nomenclature for human drug targets, where appropriate.The Concise Guide to PHARMACOLOGY 2019/20 is the fourth in this series of biennial publications. The Concise Guide provides concise overviews of the key properties of nearly 1800 human drug targets with an emphasis on selective pharmacology (where available), plus links to the open access knowledgebase source of drug targets and their ligands ( They regulate (either promoting or repressing) transcription of these target genes in response to a variety of endogenous ligands. Endogenous agonists are hydrophobic entities which, when bound to the receptor promote conformational changes in the receptor to allow recruitment (or dissociation) of protein partners, generating a large multiprotein complex.Two major subclasses of nuclear receptors with identified endogenous agonists can be identified: steroid and non\u2010steroid hormone receptors. Steroid hormone receptors function typically as dimeric entities and are thought to be resident outside the nucleus in the unliganded state in a complex with chaperone proteins, which are liberated upon agonist binding. Migration to the nucleus and interaction with other regulators of gene transcription, including RNA polymerase, acetyltransferases and deacetylases, allows gene transcription to be regulated. Non\u2010steroid hormone receptors typically exhibit a greater distribution in the nucleus in the unliganded state and interact with other nuclear receptors to form heterodimers, as well as with other regulators of gene transcription, leading to changes in gene transcription upon agonist binding.Selectivity of gene regulation is brought about through interaction of nuclear receptors with particular consensus sequences of DNA, which are arranged typically as repeats or inverted palindromes to allow accumulation of multiple transcription factors in the promoter regions of genes.NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors ) are nuclear hormone receptors of the NR1A family, with diverse roles regulating macronutrient metabolism, cognition and cardiovascular homeostasis. TRs are activated by thyroxine (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2635) and thyroid hormone (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2634). Once activated by a ligand, the receptor acts as a transcription factor either as a monomer, homodimer or heterodimer with members of the retinoid X receptor family. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2633 has been described as an antagonist at TRs with modest selectivity for TR\u03b2 [http://www.ncbi.nlm.nih.gov/pubmed/12109914?dopt=AbstractPlus].Thyroid hormone receptors Thyroid Hormone Mimetics: the Past, Current Status and Future Challenges. Curr Atheroscler Rep18: 14 https://www.ncbi.nlm.nih.gov/pubmed/26886134?dopt=AbstractPlusElbers LP et al. (2006) International Union of Pharmacology. LIX. The pharmacology and classification of the nuclear receptor superfamily: thyroid hormone receptors. Pharmacol. Rev. 58: 705\u201011 https://www.ncbi.nlm.nih.gov/pubmed/17132849?dopt=AbstractPlusFlamant F et al. (2017) New insights into thyroid hormone action. Pharmacol. Ther. 173: 135\u2010145 https://www.ncbi.nlm.nih.gov/pubmed/28174093?dopt=AbstractPlusMendoza A NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [nomenclature as agreed by the http://www.ncbi.nlm.nih.gov/pubmed/17132850?dopt=AbstractPlus]) are nuclear hormone receptors of the NR1B family activated by the vitamin A\u2010derived agonists http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2644 (ATRA) and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2645, and the RAR\u2010selective synthetic agonists http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2646 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5429. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2641 is a family\u2010selective antagonist [http://www.ncbi.nlm.nih.gov/pubmed/19477412?dopt=AbstractPlus].Retinoic acid receptors The molecular physiology of nuclear retinoic acid receptors. From health to disease. Biochim. Biophys. Acta1812: 1023\u201031 [https://www.ncbi.nlm.nih.gov/pubmed/20970498?dopt=AbstractPlus]Duong V et al. (2006) International Union of Pharmacology. LX. Retinoic acid receptors. Pharmacol. Rev. 58: 712\u201025 https://www.ncbi.nlm.nih.gov/pubmed/17132850?dopt=AbstractPlusGermain P et al. (2016) Retinoic Acid and Retinoic Acid Receptors as Pleiotropic Modulators of the Immune System. Annu. Rev. Immunol. 34: 369\u201094 https://www.ncbi.nlm.nih.gov/pubmed/27168242?dopt=AbstractPlusLarange A et al. (2017) The interrelationship between bile acid and vitamin A homeostasis. Biochim. Biophys. Acta1862: 496\u2010512 https://www.ncbi.nlm.nih.gov/pubmed/28111285?dopt=AbstractPlusSaeed A PPARs, nomenclature as agreed by theNC\u2010IUPHARSubcommittee on Nuclear Hormone Receptors [http://www.ncbi.nlm.nih.gov/pubmed/17132851?dopt=AbstractPlus]) are nuclear hormone receptors of the NR1C family, with diverse roles regulating lipid homeostasis, cellular differentiation, proliferation and the immune response. PPARs have many potential endogenous agonists , including http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1877, prostacyclin (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1915), many fatty acids and their oxidation products, lysophosphatidic acid (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2906) [http://www.ncbi.nlm.nih.gov/pubmed/12502787?dopt=AbstractPlus], http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5426, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3401, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5427, http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5428 and leukotriene B4 (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2487). http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2668 acts as a non\u2010selective agonist for the PPAR family [http://www.ncbi.nlm.nih.gov/pubmed/10691680?dopt=AbstractPlus]. These receptors also bind hypolipidaemic drugs (PPAR\u03b1) and anti\u2010diabetic thiazolidinediones (PPAR\u03b3), as well as many non\u2010steroidal anti\u2010inflammatory drugs, such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5425 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1909. Once activated by a ligand, the receptor forms a heterodimer with members of the retinoid X receptor family and can act as a transcription factor. Although radioligand binding assays have been described for all three receptors, the radioligands are not commercially available. Commonly, receptor occupancy studies are conducted using fluorescent ligands and truncated forms of the receptor limited to the ligand binding domain.Peroxisome proliferator\u2010activated receptors . Agonists with mixed activity at PPAR\u03b1 and PPAR\u03b3 have also been described .As with the estrogen receptor antagonists, many agents show tissue\u2010selective efficacy The peroxisome proliferator\u2010activated receptors in cardiovascular diseases: experimental benefits and clinical challenges. Br. J. Pharmacol. 172: 5512\u201022 [https://www.ncbi.nlm.nih.gov/pubmed/25438608?dopt=AbstractPlus]Cheang WS et al. (2017) PPARs in obesity\u2010induced T2DM, dyslipidaemia and NAFLD. Nat Rev Endocrinol13: 36\u201049 [https://www.ncbi.nlm.nih.gov/pubmed/27636730?dopt=AbstractPlus]Gross B et al. (2016) The elusive endogenous adipogenic PPAR\u03b3 agonists: Lining up the suspects. Prog. Lipid Res. 61: 149\u201062 [https://www.ncbi.nlm.nih.gov/pubmed/26703188?dopt=AbstractPlus]Hallenborg P et al. (2006) International Union of Pharmacology. LXI. Peroxisome proliferator\u2010activated receptors. Pharmacol. Rev. 58: 726\u201041 [https://www.ncbi.nlm.nih.gov/pubmed/17132851?dopt=AbstractPlus]Michalik L Trends Pharmacol. Sci. 36: 688\u2010704 [https://www.ncbi.nlm.nih.gov/pubmed/26435213?dopt=AbstractPlus]Sauer S. (2015) Ligands for the Nuclear Peroxisome Proliferator\u2010Activated Receptor Gamma. nomenclature as agreed by theNC\u2010IUPHARSubcommittee on Nuclear Hormone Receptors [http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]) have yet to be officially paired with an endogenous ligand, but are thought to be activated by heme.Rev\u2010erb receptors International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 [https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]Benoit G et al. (2015) Nuclear receptors in acute and chronic cholestasis. Dig Dis33: 357\u201066 [https://www.ncbi.nlm.nih.gov/pubmed/26045270?dopt=AbstractPlus]Gonzalez\u2010Sanchez E et al. (2015) Emerging models for the molecular basis of mammalian circadian timing. Biochemistry54: 134\u201049 [https://www.ncbi.nlm.nih.gov/pubmed/25303119?dopt=AbstractPlus]Gustafson CL et al. (2017) Drug discovery targeting heme\u2010based sensors and their coupled activities. J. Inorg. Biochem. 167: 12\u201020 [https://www.ncbi.nlm.nih.gov/pubmed/27893989?dopt=AbstractPlus]Sousa EH nomenclature as agreed by theNC\u2010IUPHARSubcommittee on Nuclear Hormone Receptors [http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]) have yet to be assigned a definitive endogenous ligand, although ROR\u03b1 may be synthesized with a \u2018captured\u2019 agonist such as http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2718 .Retinoic acid receptor\u2010related orphan receptors International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 [https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]Benoit G et al. (2016) Recent progress on nuclear receptor ROR\u03b3 modulators. Bioorg. Med. Chem. Lett. 26: 4387\u20104393 [https://www.ncbi.nlm.nih.gov/pubmed/27542308?dopt=AbstractPlus]Cyr P et al. (2006) Overview of nomenclature of nuclear receptors. Pharmacol. Rev. 58: 685\u2010704 [https://www.ncbi.nlm.nih.gov/pubmed/17132848?dopt=AbstractPlus]Germain P et al. (2016) Oxysterols in Metabolic Syndrome: From Bystander Molecules to Bioactive Lipids. Trends Mol Med22: 594\u2010614 [https://www.ncbi.nlm.nih.gov/pubmed/27286741?dopt=AbstractPlus]Guillemot\u2010Legris O et al. (2016) Oxysterols: From cholesterol metabolites to key mediators. Prog. Lipid Res. 64: 152\u2010169 [https://www.ncbi.nlm.nih.gov/pubmed/27687912?dopt=AbstractPlus]Mutemberezi V nomenclature as agreed by theNC\u2010IUPHARSubcommittee on Nuclear Hormone Receptors [http://www.ncbi.nlm.nih.gov/pubmed/17132852?dopt=AbstractPlus]) are members of a steroid analogue\u2010activated nuclear receptor subfamily, which form heterodimers with members of the retinoid X receptor family. Endogenous ligands for LXRs include hydroxycholesterols (OHC), while FXRs appear to be activated by bile acids. In humans and primates, NR1H5P is a pseudogene. However, in other mammals, it encodes a functional nuclear hormone receptor that appears to be involved in cholesterol biosynthesis [http://www.ncbi.nlm.nih.gov/pubmed/12529392?dopt=AbstractPlus].Liver X and farnesoid X receptors LXR Regulation of Brain Cholesterol: From Development to Disease. Trends Endocrinol. Metab. 27: 404\u2010414 [https://www.ncbi.nlm.nih.gov/pubmed/27113081?dopt=AbstractPlus]Courtney R et al. (2018) Recent Advances in the Medicinal Chemistry of Liver X Receptors. J. Med. Chem. 61: 10935\u201010956 [https://www.ncbi.nlm.nih.gov/pubmed/30004226?dopt=AbstractPlus]El\u2010Gendy BEM et al. (2010) Bile acids and their nuclear receptor FXR: Relevance for hepatobiliary and gastrointestinal disease. Biochim. Biophys. Acta1801: 683\u201092 [https://www.ncbi.nlm.nih.gov/pubmed/20399894?dopt=AbstractPlus]Gadaleta RM et al. (2017) Bile acids and their receptors during liver regeneration: \u201cDangerous protectors\u201d. Mol. Aspects Med. 56: 25\u201033 [https://www.ncbi.nlm.nih.gov/pubmed/28302491?dopt=AbstractPlus]Merlen G et al. (2006) International Union of Pharmacology. LXII. The NR1H and NR1I receptors: constitutive androstane receptor, pregnene X receptor, farnesoid X receptor alpha, farnesoid X receptor beta, liver X receptor alpha, liver X receptor beta, and vitamin D receptor. Pharmacol. Rev. 58: 742\u201059 [https://www.ncbi.nlm.nih.gov/pubmed/17132852?dopt=AbstractPlus]Moore DD et al. (2016) Liver X receptors: from cholesterol regulation to neuroprotection\u2010a new barrier against neurodegeneration in amyotrophic lateral sclerosis? Cell. Mol. Life Sci. 73: 3801\u20108 [https://www.ncbi.nlm.nih.gov/pubmed/27510420?dopt=AbstractPlus]Mouzat K FEBS Lett. 591: 2978\u20102991 [https://www.ncbi.nlm.nih.gov/pubmed/28555747?dopt=AbstractPlus]Schulman IG. (2017) Liver X receptors link lipid metabolism and inflammation. NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [nomenclature as agreed by the http://www.ncbi.nlm.nih.gov/pubmed/17132852?dopt=AbstractPlus]) are members of the NR1I family of nuclear receptors, which form heterodimers with members of the retinoid X receptor family. PXR and CAR are activated by a range of exogenous compounds, with no established endogenous physiological agonists, although high concentrations of bile acids and bile pigments activate PXR and CAR [http://www.ncbi.nlm.nih.gov/pubmed/17132852?dopt=AbstractPlus].Vitamin D (VDR), Pregnane X (PXR) and Constitutive Androstane (CAR) receptors International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlusBenoit G et al. (2015) Vitamin D receptor and RXR in the post\u2010genomic era. J. Cell. Physiol. 230: 758\u201066 https://www.ncbi.nlm.nih.gov/pubmed/25335912?dopt=AbstractPlusLong MD et al. (2006) International Union of Pharmacology. LXII. The NR1H and NR1I receptors: constitutive androstane receptor, pregnene X receptor, farnesoid X receptor alpha, farnesoid X receptor beta, liver X receptor alpha, liver X receptor beta, and vitamin D receptor. Pharmacol. Rev. 58: 742\u201059 https://www.ncbi.nlm.nih.gov/pubmed/17132852?dopt=AbstractPlusMoore DD NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]. While linoleic acid has been identified as the endogenous ligand for HNF4\u03b1 its function remains ambiguous [http://www.ncbi.nlm.nih.gov/pubmed/19440305?dopt=AbstractPlus]. HNF4\u03b3 has yet to be paired with an endogenous ligand.The nomenclature of hepatocyte nuclear factor\u20104 receptors is agreed by the et al. (2006) International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlusBenoit G et al. (2016) Lipid\u2010sensors, enigmatic\u2010orphan and orphan nuclear receptors as therapeutic targets in breast\u2010cancer. Oncotarget7: 42661\u201042682 https://www.ncbi.nlm.nih.gov/pubmed/26894976?dopt=AbstractPlusGarattini E et al. (2006) Overview of nomenclature of nuclear receptors. Pharmacol. Rev. 58: 685\u2010704 https://www.ncbi.nlm.nih.gov/pubmed/17132848?dopt=AbstractPlusGermain P Acta Pharm Sin B6: 393\u2010408 https://www.ncbi.nlm.nih.gov/pubmed/27709008?dopt=AbstractPlusLu H. (2016) Crosstalk of HNF4\u03b1 with extracellular and intracellular signaling pathways in the regulation of hepatic metabolism of drugs and lipids. et al. (2015) Role of hepatocyte nuclear factor 4\u03b1 (HNF4\u03b1) in cell proliferation and cancer. Gene Expr. 16: 101\u20108 https://www.ncbi.nlm.nih.gov/pubmed/25700366?dopt=AbstractPlusWalesky C NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [nomenclature as agreed by the http://www.ncbi.nlm.nih.gov/pubmed/17132853?dopt=AbstractPlus]) are NR2B family members activated by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2645 and the RXR\u2010selective agonists http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2807 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2808, sometimes referred to as rexinoids. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2816 [http://www.ncbi.nlm.nih.gov/pubmed/17947383?dopt=AbstractPlus] and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=8079 [http://www.ncbi.nlm.nih.gov/pubmed/10748721?dopt=AbstractPlus] have been described as a pan\u2010RXR antagonists. These receptors form RXR\u2010RAR heterodimers and RXR\u2010RXR homodimers .Retinoid X receptors International Union of Pharmacology. LXIII. Retinoid X receptors. Pharmacol. Rev. 58: 760\u201072 https://www.ncbi.nlm.nih.gov/pubmed/17132853?dopt=AbstractPlusGermain P et al. (2015) Vitamin D receptor and RXR in the post\u2010genomic era. J. Cell. Physiol. 230: 758\u201066 https://www.ncbi.nlm.nih.gov/pubmed/25335912?dopt=AbstractPlusLong MD et al. (2017) The multi\u2010faceted role of retinoid X receptor in bone remodeling. Cell. Mol. Life Sci. 74: 2135\u20102149 https://www.ncbi.nlm.nih.gov/pubmed/28105491?dopt=AbstractPlusMen\u00e9ndez\u2010Guti\u00e9rrez MP NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [nomenclature as agreed by the http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]) have yet to be officially paired with an endogenous ligand, although testicular receptor 4 has been reported to respond to retinoids.Testicular receptors International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlusBenoit G et al. (2006) Overview of nomenclature of nuclear receptors. Pharmacol. Rev. 58: 685\u2010704 https://www.ncbi.nlm.nih.gov/pubmed/17132848?dopt=AbstractPlusGermain P et al. (2014) Minireview: role of orphan nuclear receptors in cancer and potential as drug targets. Mol. Endocrinol. 28: 157\u201072 https://www.ncbi.nlm.nih.gov/pubmed/24295738?dopt=AbstractPlusSafe S et al. (2016) The emerging roles of orphan nuclear receptors in prostate cancer. Biochim. Biophys. Acta1866: 23\u201036 https://www.ncbi.nlm.nih.gov/pubmed/27264242?dopt=AbstractPlusWu D NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [nomenclature as agreed by the http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]) have yet to be officially paired with an endogenous ligand.Tailless\u2010like receptors TLX: An elusive receptor. J. Steroid Biochem. Mol. Biol. 157: 41\u20107 https://www.ncbi.nlm.nih.gov/pubmed/26554934?dopt=AbstractPlusBenod C et al. (2006) International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlusBenoit G et al. (2006) Overview of nomenclature of nuclear receptors. Pharmacol. Rev. 58: 685\u2010704 https://www.ncbi.nlm.nih.gov/pubmed/17132848?dopt=AbstractPlusGermain P et al. (2018) Regulation of behaviour by the nuclear receptor TLX. Genes Brain Behav. 17: e12357 https://www.ncbi.nlm.nih.gov/pubmed/27790850?dopt=AbstractPlusO\u2019Leary JD NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [nomenclature as agreed by the http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]) have yet to be officially paired with an endogenous ligand.COUP\u2010TF\u2010like receptors International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlusBenoit G et al. (2006) Overview of nomenclature of nuclear receptors. Pharmacol. Rev. 58: 685\u2010704 https://www.ncbi.nlm.nih.gov/pubmed/17132848?dopt=AbstractPlusGermain P et al. (2016) The emerging roles of orphan nuclear receptors in prostate cancer. Biochim. Biophys. Acta1866: 23\u201036 https://www.ncbi.nlm.nih.gov/pubmed/27264242?dopt=AbstractPlusWu D etal. (2016) Choose your destiny: Make a cell fate decision with COUP\u2010TFII. J. Steroid Biochem. Mol. Biol. 157: 7\u201012 https://www.ncbi.nlm.nih.gov/pubmed/26658017?dopt=AbstractPlusWu SP NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [nomenclature as agreed by the http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]) have yet to be officially paired with an endogenous ligand.Estrogen\u2010related receptors International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlusBenoit G et al. (2016) Estrogen\u2010related receptor \u03b2 (ERR\u03b2) \u2010 renaissance receptor or receptor renaissance? Nucl Recept Signal14: e002 https://www.ncbi.nlm.nih.gov/pubmed/27507929?dopt=AbstractPlusDivekar SD et al. (2006) Overview of nomenclature of nuclear receptors. Pharmacol. Rev. 58: 685\u2010704 https://www.ncbi.nlm.nih.gov/pubmed/17132848?dopt=AbstractPlusGermain P et al. (2016) There and back again: The journey of the estrogen\u2010related receptors in the cancer realm. J. Steroid Biochem. Mol. Biol. 157: 13\u20109 https://www.ncbi.nlm.nih.gov/pubmed/26151739?dopt=AbstractPlusTam IS et al. (2016) The emerging roles of orphan nuclear receptors in prostate cancer. Biochim. Biophys. Acta1866: 23\u201036 https://www.ncbi.nlm.nih.gov/pubmed/27264242?dopt=AbstractPlusWu D NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [nomenclature as agreed by the http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]) have yet to be officially paired with an endogenous ligand.Nerve growth factor IB\u2010like receptors International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlusBenoit G etal. (2006) Overview of nomenclature of nuclear receptors. Pharmacol. Rev. 58: 685\u2010704 https://www.ncbi.nlm.nih.gov/pubmed/17132848?dopt=AbstractPlusGermain PJ. Recept. Signal Transduct. Res. 35: 184\u20108 https://www.ncbi.nlm.nih.gov/pubmed/25089663?dopt=AbstractPlusRanhotra HS. (2015) The NR4A orphan nuclear receptors: mediators in metabolism and diseases. et al. (2017) The NR4A subfamily of nuclear receptors: potential new therapeutic targets for the treatment of inflammatory diseases. Expert Opin. Ther. Targets21: 291\u2010304 https://www.ncbi.nlm.nih.gov/pubmed/28055275?dopt=AbstractPlusRodr\u00edguez\u2010Calvo R et al. (2016) Nuclear receptor 4A (NR4A) family \u2010 orphans no more. J. Steroid Biochem. Mol. Biol. 157: 48\u201060 https://www.ncbi.nlm.nih.gov/pubmed/25917081?dopt=AbstractPlusSafe S NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [nomenclature as agreed by the http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]) have yet to be officially paired with an endogenous ligand.Fushi tarazu F1\u2010like receptors International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlusBenoit G et al. (2016) Lipid\u2010sensors, enigmatic\u2010orphan and orphan nuclear receptors as therapeutic targets in breast\u2010cancer. Oncotarget7: 42661\u201042682 https://www.ncbi.nlm.nih.gov/pubmed/26894976?dopt=AbstractPlusGarattini E et al. (2006) Overview of nomenclature of nuclear receptors. Pharmacol. Rev. 58: 685\u2010704 https://www.ncbi.nlm.nih.gov/pubmed/17132848?dopt=AbstractPlusGermain P et al. (2016) Structures and regulation of non\u2010X orphan nuclear receptors: A retinoid hypothesis. J. Steroid Biochem. Mol. Biol. 157: 27\u201040 https://www.ncbi.nlm.nih.gov/pubmed/26159912?dopt=AbstractPlusZhi X et al. (2015) Nuclear receptor variants in liver disease. Dig Dis33: 415\u20109 https://www.ncbi.nlm.nih.gov/pubmed/26045277?dopt=AbstractPlusZimmer V NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors [nomenclature as agreed by the http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]) have yet to be officially paired with an endogenous ligand.Germ cell nuclear factor receptors International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlusBenoit G et al. (2016) Lipid\u2010sensors, enigmatic\u2010orphan and orphan nuclear receptors as therapeutic in breast\u2010cancer. Oncotarget7: 42661\u201042682 https://www.ncbi.nlm.nih.gov/pubmed/26894976?dopt=AbstractPlusGarattini E et al. (2006) Overview of nomenclature of nuclear receptors. Pharmacol. Rev. 58: 685\u2010704 https://www.ncbi.nlm.nih.gov/pubmed/17132848?dopt=AbstractPlusGermain P et al. (2014) Minireview: role of orphan nuclear receptors in cancer and potential as drug Mol. Endocrinol. 28: 157\u201072 https://www.ncbi.nlm.nih.gov/pubmed/24295738?dopt=AbstractPlusSafe S et al. (2016) Structures and regulation of non\u2010X orphan nuclear receptors: A retinoid hypothesis. J. Steroid Biochem. Mol. Biol. 157: 27\u201040 https://www.ncbi.nlm.nih.gov/pubmed/26159912?dopt=AbstractPlusZhi X nomenclature as agreed by theNC\u2010IUPHARSubcommittee on Nuclear Hormone Receptors [http://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlus]) have yet to be officially paired with an endogenous ligand.Dax\u2010like receptors International Union of Pharmacology. LXVI. Orphan nuclear receptors. Pharmacol. Rev. 58: 798\u2010836 https://www.ncbi.nlm.nih.gov/pubmed/17132856?dopt=AbstractPlusBenoit G et al. (2016) Lipid\u2010sensors, enigmatic\u2010orphan and orphan nuclear receptors as therapeutic targets in breast\u2010cancer. Oncotarget7: 42661\u201042682 https://www.ncbi.nlm.nih.gov/pubmed/26894976?dopt=AbstractPlusGarattini E et al. (2006) Overview of nomenclature of nuclear receptors. Pharmacol. Rev. 58: 685\u2010704 https://www.ncbi.nlm.nih.gov/pubmed/17132848?dopt=AbstractPlusGermain P et al. (2014) Minireview: role of orphan nuclear receptors in cancer and potential as drug targets. Mol. Endocrinol. 28: 157\u201072 https://www.ncbi.nlm.nih.gov/pubmed/24295738?dopt=AbstractPlusSafe S et al. (2016) The emerging roles of orphan nuclear receptors in prostate cancer. Biochim. Biophys. Acta1866: 23\u201036 https://www.ncbi.nlm.nih.gov/pubmed/27264242?dopt=AbstractPlusWu D NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors ) are nuclear hormone receptors of the NR3 class, with endogenous agonists that may be divided into 3\u2010hydroxysteroids (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2818 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1013) and 3\u2010ketosteroids . These receptors exist as dimers coupled with chaperone molecules and immunophilin FKBP52:https://www.genenames.org/data/gene\u2010symbol\u2010report/#!/hgnc_id/HGNC:3720, http://www.uniprot.org/uniprot/Q02790), which are shed on binding the steroid hormone. Although rapid signalling phenomena are observed , the principal signalling cascade appears to involve binding of the activated receptors to nuclear hormone response elements of the genome, with a 15\u2010nucleotide consensus sequence AGAACAnnnTGTTCT as homo\u2010 or heterodimers. They also affect transcription by protein\u2010protein interactions with other transcription factors, such as activator protein 1 (AP\u20101) and nuclear factor \u03baB (NF\u2010\u03baB). Splice variants of each of these receptors can form functional or non\u2010functional monomers that can dimerize to form functional or non\u2010functional receptors. For example, alternative splicing of PR mRNA produces A and B monomers that combine to produce functional AA, AB and BB receptors with distinct characteristics [http://www.ncbi.nlm.nih.gov/pubmed/8264658?dopt=AbstractPlus].Steroid hormone receptors has been described. Human orthologues of 7TM \u2019membrane progestin receptors\u2019 , initially discovered in fish , appear to localize to intracellular membranes and respond to \u2019non\u2010genomic\u2019 progesterone analogues independently of G proteins [http://www.ncbi.nlm.nih.gov/pubmed/18603275?dopt=AbstractPlus].A 7TM receptor responsive to estrogen activity regulates diverse physiological processes http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2822 exhibits partial agonist activity at ER\u03b1 . Estrogen receptors may be blocked non\u2010selectively by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1016 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2820 and labelled by http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1012 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5384. Many agents thought initially to be antagonists at estrogen receptors appear to have tissue\u2010specific efficacy , hence the descriptor SERM (selective estrogen receptor modulator) or SnuRM (selective nuclear receptor modulator). http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=5430 has been suggested to be an ER\u03b1\u2010selective estrogen receptor modulator [http://www.ncbi.nlm.nih.gov/pubmed/17115070?dopt=AbstractPlus].et al. (2017) DNA Sequence Constraints Define Functionally Active Steroid Nuclear Receptor Binding Sites in Chromatin. Endocrinology158: 3212\u20103234 https://www.ncbi.nlm.nih.gov/pubmed/28977594?dopt=AbstractPlusCoons LA et al. (2006) International Union of Pharmacology. LXIV. Estrogen receptors. Pharmacol. Rev. 58: 773\u201081 https://www.ncbi.nlm.nih.gov/pubmed/17132854?dopt=AbstractPlusDahlman\u2010Wright K et al. (2015) Nuclear receptors in acute and chronic cholestasis. Dig Dis33: 357\u201066 https://www.ncbi.nlm.nih.gov/pubmed/26045270?dopt=AbstractPlusGonzalez\u2010Sanchez E et al. (2016) What\u2019s new in estrogen receptor action in the female reproductive tract. J. Mol. Endocrinol. 56: R55\u201071 https://www.ncbi.nlm.nih.gov/pubmed/26826253?dopt=AbstractPlusHewitt SC et al. (2017) Estrogen receptor agonists/antagonists in breast cancer therapy: A critical review. Bioorg. Chem. 71: 257\u2010274 https://www.ncbi.nlm.nih.gov/pubmed/28274582?dopt=AbstractPlusJameera Begam A et al. (2017) Estrogen Receptor \u03b2 as a Pharmaceutical Target. Trends Pharmacol. Sci. 38: 92\u201099 https://www.ncbi.nlm.nih.gov/pubmed/27979317?dopt=AbstractPlusWarner M NC\u2010IUPHAR Subcommittee on Nuclear Hormone Receptors ) are nuclear hormone receptors of the NR3 class, with endogenous agonists that may be divided into 3\u2010hydroxysteroids (http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=2818 and http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=1013) and 3\u2010ketosteroids .Steroid hormone receptors and Type II (e.g. http://www.guidetopharmacology.org/GRAC/LigandDisplayForward?ligandId=3448) groups. These groups appear to promote binding of PR to DNA with different efficacies and evoke distinct conformational changes in the receptor, leading to a transcription\u2010neutral complex . Mutations in AR underlie testicular feminization and androgen insensitivity syndromes, spinal and bulbar muscular atrophy (Kennedy\u2019s disease).et al. (2017) 30 YEARS OF THE MINERALOCORTICOID RECEPTOR: Evolution of the mineralocorticoid receptor: sequence, structure and function. J. Endocrinol. 234: T1\u2010T16 https://www.ncbi.nlm.nih.gov/pubmed/28468932?dopt=AbstractPlusBaker ME et al. (2017) Deciphering the divergent roles of progestogens in breast cancer. Nat. Rev. Cancer17: 54\u201064 https://www.ncbi.nlm.nih.gov/pubmed/27885264?dopt=AbstractPlusCarroll JS et al. (2017) Nuclear Receptor Function through Genomics: Lessons from the Glucocorticoid Receptor. Trends Endocrinol. Metab. 28: 531\u2010540 https://www.ncbi.nlm.nih.gov/pubmed/28495406?dopt=AbstractPlusCohen DM et al. (2017) Brain mineralocorticoid receptor function in control of salt balance and stress\u2010adaptation. Physiol. Behav. 178: 13\u201020 https://www.ncbi.nlm.nih.gov/pubmed/28089704?dopt=AbstractPlusde Kloet ER et al. (2017) Progesterone\u2010Mediated Non\u2010Classical Signaling. Trends Endocrinol. Metab. 28: 656\u2010668 https://www.ncbi.nlm.nih.gov/pubmed/28651856?dopt=AbstractPlusGarg D et al. (2006) International Union of Pharmacology. LXV. The pharmacology and classification of the nuclear receptor superfamily: glucocorticoid, mineralocorticoid, progesterone, and androgen receptors. Pharmacol. Rev. 58: 782\u201097 https://www.ncbi.nlm.nih.gov/pubmed/17132855?dopt=AbstractPlusLu NZ et al. (2017) Genomic and non\u2010genomic effects of androgens in the cardiovascular system: clinical implications. Clin. Sci. 131: 1405\u20101418 https://www.ncbi.nlm.nih.gov/pubmed/28645930?dopt=AbstractPlusLucas\u2010Herald AK et al. (2017) Androgen receptor splice variants and prostate cancer: From bench to bedside. Oncotarget8: 18550\u201018576 https://www.ncbi.nlm.nih.gov/pubmed/28077788?dopt=AbstractPlusWadosky KM et al. (2017) Glucocorticoid receptor control of transcription: precision and plasticity via allostery. Nat. Rev. Mol. Cell Biol. 18: 159\u2010174 https://www.ncbi.nlm.nih.gov/pubmed/28053348?dopt=AbstractPlusWeikum ER"} +{"text": "The Pan African Medical Journal. 2016;24:335. doi:10.11604/pamj.2016.24.335.9171.This erratum corrects article: The original version of this article had the"} +{"text": "This article has been corrected: Due to errors in image placement, the representative image of comet assay in group HT29 shG9a for 2917-2927. https://doi.org/10.18632/oncotarget.2784Original article: Oncotarget. 2015; 6:2917\u20132927."} +{"text": "This article has been corrected: The correct reference 5 is given below:5. Berg JK, et al. Circulation Heart Failure. 2017; 10.106165-106166. https://doi.org/10.18632/oncotarget.22579Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: The correct author name is given below:Michela Piezzo31877-31887. https://doi.org/10.18632/oncotarget.25874Original article: Oncotarget. 2018; 9:"} +{"text": "This article has been corrected: Due to an accidental duplication, the images for Grade I MMP-2 staining and Normal Cartilage Resistin staining in Figure https://doi.org/10.18632/oncotarget.2724Original article: Oncotarget. 2015; 6:258-270."} +{"text": "Clostridioides was misspelled in Risk for Clostridioides difficile Infection among Older Adults with Cancer . The article has been corrected online (https://wwwnc.cdc.gov/eid/article/25/9/18-1142_article)."} +{"text": "Correction of \u201cSteven, L. L., M.F. Jennifer, and P.U. Johua. 2019. Assessing the Potential for Interaction in Insecticidal Activity Between MON 87751 \u00d7 MON 87701 Produced by Conventional Breeding. Environ. Entomol.\u201d10.1093/ee/nvz082Doi: This article incorrectly displayed an author name online. The author\u2019s correct name is Joshua P. Uffman. This has been corrected online and in print. The authors regret this error."} +{"text": "This article has been corrected: Due to errors in figure preparation, the Western ink point p-Akt protein was accidentally used in the wrong picture in 58915-58930. https://doi.org/10.18632/oncotarget.10410Original article: Oncotarget. 2016; 7:58915\u201358930."} +{"text": "This article has been corrected: The correct spelling of author name is given below:Shaojin Zhang77942-77956. https://doi.org/10.18632/oncotarget.18549Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: The correct author name is given below:Claudio Di Cristofano30624-30634. https://doi.org/10.18632/oncotarget.25755Original article: Oncotarget. 2018; 9:"} +{"text": "This article has been corrected: The correct author name is given below:Jung Ki Johttps://doi.org/10.18632/oncotarget.18298Original article: Oncotarget. 2017; 8:96893-96902."} +{"text": "This article has been corrected: The correct funding information is given below:FUNDINGThis work was in part supported by German Cancer Foundation (Grant no. 70112372), German Research Foundation (SCHU 2681/1-1), HERA Women's Cancer Foundation OSB1 Grant (C.S.) and the National Institutes of Health , TEDCO/Maryland Innovation Initiative and Neximmune, Inc. MD Biotech Center (J.P.S.).68503-68512. https://doi.org/10.18632/oncotarget.11785Original article: Oncotarget. 2016; 7:"} +{"text": "This article has been corrected: During production, the ending page number for this article was listed incorrectly. The page count has now been adjusted to show the proper pagination.362-372. https://doi.org/10.18632/oncotarget.901Original article: Oncotarget. 2012; 4:"} +{"text": "This article has been corrected: The updated acknowledgements are given below:ACKNOWLEDGMENTSWe thank Dr. Raffaella Sordella from Cold Spring Harbor Laboratory for kindly providing the valuable cell lines used in the current study. We thank Mrs. Juan Du for taking immunofluoresence images. The work in Zhao Lab was supported by AFOSR grant no. FA9550-16- 1-0052.95741-95754. https://doi.org/10.18632/oncotarget.21306Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: The correct author name is given below:Cheryl A. London22693-22702. https://doi.org/10.18632/oncotarget.25209Original article: Oncotarget. 2018; 9:"} +{"text": "The Pan African Medical Journal. 2017;28:307. doi:10.11604/pamj.2017.28.307.14507.This corrigendum corrects article The original version of this article had the"} +{"text": "Healthcare-associated infections and antibiotic resistanceMeticillin-resistant Staphylococcus aureus 2016EPI-NEWS 23, 20177 June 2017, Denmarkhttp://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2023%20-%202017.aspxKlebsiella spp. bacteraemia in England, Wales and Northern Ireland: 2016Laboratory surveillance of Health Protection Report; 11(18)22 May 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/615375/hpr1817_klbsll.pdfEscherichia coli bacteraemia in England, Wales and Northern Ireland: 2016 Health Protection Report; 11(18)Laboratory surveillance of 22 May 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/615340/hpr1817_ecoli.pdfEnterococcus spp. bacteraemia in England, Wales and Northern Ireland: 2016 Health Protection Report; 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11(18)12 May 2017, United Kingdomhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/613906/hpr1717_SF-GAS.pdfUnraveling a tuberculosis clusterVlaams Infectieziektebulletin; 1/2017April 2017, Belgiumhttps://www.zorg-en-gezondheid.be/sites/default/files/atoms/files/tb-ontrafeling-C.dehollogne.pdfContact tracing after the death of a patient with XDR-tuberculosis on an airplane, Germany 2013Epidemiologisches Bulletin 13, 201730 March 2017, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2017/Ausgaben/13_17.pdf?__blob=publicationFileSexually transmitted diseasesSyphilis 2016EPI-NEWS 21/22, 201731 May 2017, Denmarkhttp://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2021-22%20-%202017.aspxGonorrhoea 2016EPI-NEWS 18, 20173 May 2017, Denmarkhttp://www.ssi.dk/Aktuelt/Nyhedsbreve/EPI-NYT/2017/Uge%2018%20-%202017.aspxSexually transmitted diseasesEpi-Ice 2017;10(2)April 2017, Icelandhttp://www.landlaeknir.is/servlet/file/store93/item32291/EPI-ICE_April_2017.pdfSexually transmitted infections and HIV in Ireland, 2016: provisional dataEpi-Insight 2017;18(4)April 2017, Irelandhttp://ndsc.newsweaver.ie/epiinsight/s9b8h9tdfmi10gkzp9yxn5?a=2&p=51673455&t=17517804Zoonoses and vector-borne diseases Surveillance of chikungunya, dengue and Zika virus infection in mainland France, 2016Bulletin \u00e9pid\u00e9miologique hebdomadaire, 1230 May 2017, Francehttp://invs.santepubliquefrance.fr/beh/2017/12/pdf/2017_12.pdfZoonotic infections with Mycobacterium tuberculosis in German livestockEpidemiologisches Bulletin 20, 201718 May 2017, Germanyhttps://www.rki.de/DE/Content/Infekt/EpidBull/Archiv/2017/Ausgaben/20_17.pdf?__blob=publicationFileCommon animal associated infections quarterly report \u2013 first quarter 2017 Health Protection Report; 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51(18)9 May 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1718.pdfTRAVAX Outbreak Index 2016: Diseases of Public and Travel Health RiskHPS Weekly Report 2017; 51(17)2 May 2017, Scotlandhttp://www.hps.scot.nhs.uk/documents/ewr/pdf2017/1717.pdf"} +{"text": "This article has been corrected: The correct author name is given below:Alfredo Minguelahttps://doi.org/10.18632/oncotarget.16657Original article: Oncotarget. 2017; 8:31959-31976."} +{"text": "The uro<\u20090.001) . An expl<\u20090.001) . In the <\u20090.001) . Continu<\u20090.001) . New hig<\u20090.001) . This ca<\u20090.001) . To date"} +{"text": "This article has been corrected: The correct author affiliation information is given below:1,2Hong Zhao93813-93824. https://doi.org/10.18632/oncotarget.21217Original article: Oncotarget. 2017; 8:"} +{"text": "This article has been corrected: Due to errors during processing, 1231-1248. https://doi.org/10.18632/oncotarget.2840Original article: Oncotarget. 2015; 6:1231\u20131248."} +{"text": "Psychologica Belgica. 57(1), pp.32\u201342. DOI: https://doi.org/10.5334/pb.348This article details a correction to the article: Verschueren, M., et al., . Identit The correctFigure The original Figure 2017"} +{"text": "In the Funding section, the grant number from the funder Wellcome Trust/MRC is listed incorrectly. The correct grant number is: 089703/Z/09/Z, 099133/Z/12/Z."} +{"text": "This article has been corrected: Due to errors during figure preparation, the images for 3800-3812. https://doi.org/10.18632/oncotarget.1998Original article: Oncotarget. 2014; 5:3800\u20133812."} +{"text": "Correction to: BMC Med Educ (2019) 19:402https://doi.org/10.1186/s12909-019-1841-2Following publication of the original article , due to Page 1 Correspondence.Incorrect: * Correspondence: Zaid.Imam@beaumont.org; Mitchell.Cappell@beaumont.eduCorrect:*Correspondence: Mitchell.Cappell@beaumont.eduPage 2 Methods (paragraph 1).Incorrect: for endoscopy retrograde.Correct: for endoscopic retrograde.Page 3 Statistical analysis (paragraph 1).Incorrect: years of (instead.Correct: years (instead.Incorrect: year) interviews.Correct: year) of interviewees.Page 3 Results (Significantly varying parameters with time).Incorrect: SimplePara>Fig. 1a illustrates.Correct: Fig. 1a illustrates.Page 4 Table 1.Incorrect: Is missing a row in the middle of the table which should be in the table as a second header.Correct: The corrected Table Page 5 Bottom, right columnIncorrect: 2009 and 2011 vs.2016\u20132018.Correct: 2009\u20132011 vs. 2016\u20132018.Page 6 Table 3.Incorrect: It should have 3 columns and not 1 column.Correct: The corrected Table Page 6 Under Table 3 (paragraph 4)Incorrect: are.Correct: were.Incorrect: ten-year-study.Correct: ten-year-study-period.Page 7 Author\u2019s contributions.Incorrect: as mentor for Dr. ZI. Both authors read and approved the final manuscript.Correct: as mentor for Dr. ZI. Both authors are equal and primary authors. Both authors read and approved the final manuscript.Page 7 Competing interests.Incorrect: Gastroenterology.Correct: Gastrointestinal."} +{"text": "Nat. Commun. 10.1038/s41467-018-06036-0; published online 6 Sep 2018.Correction to: https://www.ams.ethz.ch/research.html.\u2019 The correct version states \u2018http://www.ams.ethz.ch/research/published-data.html\u2019 in place of \u2018https://www.ams.ethz.ch/research.html\u2019. This has been corrected in both the PDF and HTML versions of the Article.The original version of this Article contained an error in the Data Availability section, which incorrectly read \u2018All data will be freely available via"} +{"text": "Correction to: J Biomed Scihttps://doi.org/10.1186/s12929-018-0481-xWNT1 p.Gly324Cys should be c.970G>T instead of c.1168G>T.In the original publication of this article , there a"} +{"text": "BJC would like to extend our sincere thanks to the referees for contributing their expertise and time.In this issue, we publish the names of those who reviewed the manuscripts for us in 2017. The Editor-in-Chief, Subject Editors and everyone involved in publishing Philippe A. CassierLauri A. AaltonenMohamed Abdel-RahmanEric O. AboagyeThomas AbramsAlessio AccardiMarc G. AchenAnya AdairPeter AdamsKasper AdelborgNabil AdraPrasad S. AdusumilliIlir AgalliuLars Agr\u00e9usM. Teresa Agull\u00f3-Ortu\u00f1oThomas AhernNihal AhmadAtique U. AhmedTim AitmanJaffer A. AjaniR. A. Al-NaggarTaleb H. Al-TelCostantine AlbanySimak AliEmma H. AllottMaria AlsinaPeter AltevogtRosario AmatoSrikanth R. AmbatiPeter F. AmbrosMaria R. AmbrosioBilly AmzalSudarshan AnandCarey K. AndersAnnie AndersonKristin E. AndersonFabrice Andr\u00e9Nicolas Andr\u00e9Gabriella AndreottiZhiwei AngRoberto AngioliAndrea AnichiniHendrik Jan AnkersmitChristina M. AnnunziataAlan AnthoneyMichael H. AntoniStefan AntonowiczAnnika AntonssonMikko AnttonenFawzi AoudjitThomas AparicioGiuseppe AprileRami I. AqeilanD. J. ArgyleHendrik-Tobias ArkenauGregory T. ArmstrongRudolf ArnoldSidsel ArnsprangGrazia ArpinoNuri ArslanBanu ArunJohan AsklingIgor AstsaturovKyrgidis AthanassiosM Hammad AtherChloe AtreyaFederico N. AucejoFran\u00e7ois AudenetLeonard AugenlichtMichael J. AusserlechnerAdnan AydinerS. C. AzouryReinhard B\u00fcttnerHideo BabaJean-Baptiste BachetRajendra BadweCatherine BagotDavid BajorShairaz BakshGustavo BaldassarreClaudia R. BallCynthia BamdadUdai BanerjiShamila A. 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WhiteArmin WiegeringJoseph WiemelsHans WildiersAnna Clare WilkinsHolger WillenbergWalter WillettJohn A. WilliamsAnna-Lise WilliamsonStuart Charles WilliamsonWilliam R. WilsonD. M. WinnDes WinterKerri M. Winters-StonePenella J. WollC. C. WongStephen Q. WongCameron WrightWoodring WrightQijun WuXiaohua WuYihua WuW. WulaningsihWahyu WulaningsihLynda WyldWei XiaoShao-An XueLusine YaghjyanKrishna YallaTesshi YamadaMasayuki YamamotoMotohisa YamamotoNoboru YamamotoKatsuhiko YanagaChih-Hsin YangTimothy A. YapLinda S. YasuiLucy YatesJohn YaxleyTsz Lun YeungMin YiHoward YimZhang YingjianDesmond YipKazuhiro YoshidaCaroline YoungPaul W. YoungGeorge M. YousefKenneth H. YuYing YuanMathew YurgelunNadia ZaffaroniNousheen ZaidiF. ZalazarJohn ZalcbergKhalil ZamanRita ZamarchiXingxing ZangA. G. ZeimetYoh ZenJincheng ZengShan ZengDonna ZhangJinfeng ZhangLi ZhangShawn (Xiang) ZhangShutian ZhangXiaodong ZhangXiaowen ZhangXinhua ZhangYi ZhangZheng-Yun ZhangZhiqian ZhangZuo-Feng ZhangPeiming ZhengDawang ZhouGuanglei ZhuangAlessandra ZingoniZhao Zuowei"} +{"text": "This article has been corrected: The correct Author name is given below:Soren Hayrabedyanhttps://doi.org/10.18632/oncotarget.16028Original article: Oncotarget. 2017; 8:32419-32432."} +{"text": "While not members of the Board of Editors, invited editors serve an important role in the review process. Invited editors are experts in their fields of research who add an additional level of quality to the review process. An editor may assign a paper to an invited editor when he/she would like to have an additional expert opinion of the reviews or when the subject area falls outside the editor\u2019s primary area of expertise.On behalf of the editors of John M. ArchibaldMartin Fabian BachmannFabio BagnoliVanessa BaileyFernando BaqueroSonia L. BardyFrederic J. BarrasThomas BernhardtElisabeth M. BikSteven R. BlankeJesse D. BloomDaniel R. BondJoe Bondy-DenomyIvo Gomperts BonecaJohn Dallas BoycePatricia A. BradfordAxel A. BrakhageStephane BretagneWilliam J. BrittMichael A. BrockhurstPierre A. BuffetSwaine L. ChenJohn M. CoffinStephen ColemanVaughn S. CooperThelka CordesFevzi DaldalJames B. DaleBlossom DamaniaDenise DearingTanneke den BlaauwenEric Y. DenkersLars E. P. DietrichStephen P. DiggleDaniel DiMaioMaxwell DowNisha DuggalLothar EllingJorge C. Escalante-SemerenaAriberto FassatiEdward J. FeilAlain FillouxWoodward W. FischerSuzanne M. J. FleiszigClaire M. FraserClay FuquaJorge E. GalanRobert F. GarryMaria Laura GennaroMimi GhoshDeanna L. GibsonSteven R. GillBenjamin GlickVernita GordonSabina GorskaHeinrich Georg GottlingerJean GruenbergAngelika Gr\u00fcndlingHenk P. HaagsmanMaria HadjifrangiskouSusan HafensteinNeal D. HammerDavid HarrichRasika HarsheyThomas HawnGerald L. HazelbauerSophie HelaineAndrew J. HendersonHubert HilbiDeborah A. HoganKerwyn Casey HuangKelly T. HughesDavid A. HunstadUrs JenalDavid KadoshGanjam V. KalpanaFatah KashanchiDaniel B. KearnsBrian KelsallBruce S. KleinManuel KleinerTheresa M. KoehlerJulia Ruth K\u00f6hlerEugene V. KooninPeter KraiczyOscar P. KuipersKim LewisShan-Lu LiuJoseph LutkenhausFrank MaldarelliHarmit S. MalikDavid M. MargolisAdam MartinyLeonard MindichHarry L. T. MobleyKarl M\u00fcngerKenneth H. NealsonJames D. OliverBeth OrcuttEric OswaldMelanie OttBernhard O. PalssonLaila Partida-MartinezThomas PietschmannAlexander PlossPaul B. RaineyTimothy D. ReadDavid A. RelmanJyothi RengarajanKyu Young RheeAndrew RiceJason W. RoschGian Maria RossoliniMonica J. RothManish SagarJason W. SahlLinda J. Saif\u00c1lvaro San MillanSaumendra N. SarkarKarla J. F. SatchellJeffrey W. SchertzerBarbara ShacklettWilliam M. ShaferLilach SheinerDavid R. ShermanChiaho ShihRobert F. SilicianoViviana SimonEric P. SkaarDavid SkurnikMark S. SmeltzerDavid G. ThanassiAlfredo G. TorresAna TravenM. Stephen TrentRodney K. TwetenSusana T. ValenteHenny C. van der MeiWillem Van SchaikDavid WangScott C. WeaverRobert T. WheelerAlex WilsonHui WuJae-Hyuk YuEgija ZauraZhi-Ming ZhengJingen ZhuThe time and effort of the following experts in handling articles have been essential to ensuring the high quality of our publications, and their help is greatly appreciated."} +{"text": "This article has been corrected: The correct email for correspondence author is given below:Correspondence to: Eduardo Nagore,email:eduardo.nagore@ucv.eshttps://doi.org/10.18632/oncotarget.22016Original article: Oncotarget. 2017; 8:98876-98886."} +{"text": "P\u2009<\u2009.0001) [We read the study by Thomas-R\u00fcddel et al. with great interest . Authors<\u2009.0001) . As PCT <\u2009.0001) , the con<\u2009.0001) . Also, u<\u2009.0001) . Accordi<\u2009.0001) ."} +{"text": "This article has been corrected: Due to errors in image preparation, the separate red/green images presented for vehicle control and valproate in 90262-90277 . https://doi.org/10.18632/oncotarget.21660Original article: Oncotarget. 2017; 8:90262\u201390277."} +{"text": "Leber congenital amaurosis/early-onset severe retinal dystrophy: clinical features, molecular genetics and therapeutic interventions. Br J Ophthalmol 2017;101:1147\u201354. doi: 10.1136/bjophthalmol-2016-309975.Kumaran N, Moore AT, Weleber RG, The authors wish to correct the legend of table 1. It reads: *Genes associated with EOSRD. \u2020Genes more frequently associated with LCA. However it should read: \u2020Genes associated with EOSRD. *Genes more frequently associated with LCA."} +{"text": "This article has been corrected: Due to errors in image processing, an IHC staining slide of ER in 87194-87208. https://doi.org/10.18632/oncotarget.19909 Original article: Oncotarget. 2017; 8:87194\u201387208."} +{"text": "This article has been corrected: The new Correspondence details are given below:1Bingjin LiCorrespondence to: Bingjin Li, email:libingjin@jlu.edu.cn78225-78233. https://doi.org/10.18632/oncotarget.20606Original article: Oncotarget. 2017; 8:78225\u201378233."} +{"text": "Rev. Saude Publica [online]. 2019, vol.53:52, ISSN 1518-8787, http://dx.doi.org/10.11606/s1518-8787.2019053000909, the RSP corrects the author\u2019s last name.In Where you read:Daniela Saes SarotelliYou should read:Daniela Saes Sartorelli \u201cPadr\u00f5es alimentares de gestantes, excesso de peso materno e diabetes gestacional\u201d. Rev. Saude Publica [online]. 2019, vol.53:52, ISSN 1518-8787, http://dx.doi.org/10.11606/s1518-8787.2019053000909, a RSP corrige o sobrenome do autor.No artigo Onde se l\u00ea:Daniela Saes SarotelliLeia-se:Daniela Saes Sartorelli"} +{"text": "Nature Communications8: Article number: 15857; DOI: 10.1038/ncomms15857 (2017); Published 07042017; Updated 11192018.This Article contains an error in reference 1. The correct reference is as follows:Nat. Cell Biol.18, 711\u2013717 (2016).S\u00e1nchez, I. & Dynlacht, B. D. Cilium assembly and disassembly."} +{"text": "Correction to: Trials (2019) 20:49.https://doi.org/10.1186/s13063-018-3169-3Incorrect name tagging:Last Name: Pasqua.First Name: Oscar Della.Correct author name tagging:Last Name: Della Pasqua.First Name: Oscar.Following publication of the original article , we have"} +{"text": "Trypanosoma cruzi-specific IFN-\u03b3-producing cells in chronic Chagas disease associate with a functional IL-7/IL-7R axis. PLoS Negl Trop Dis 12(12): e0006998. https://doi.org/10.1371/journal.pntd.0006998The second author's name is spelled incorrectly. The correct name is: Gonzalo Cesar. The correct citation is: Natale MA, Cesar G, Alvarez MG, Castro Eiro MD, Lococo B, Bertocchi G, et al. 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Peptides are key in curbing virus transmission, and cyclic peptides have a greater survival time than their linear peptides. To date, the accurate prediction of cyclic peptide-protein receptors binding conformations still is challenging because of high peptide flexibility. Here, a useful approach combined the global peptide docking, Gaussian accelerated molecular dynamics (GaMD), two-dimensional (2D) potential of mean force (PMF), normal molecular dynamics (cMD), and solvated interaction energy (SIE) techniques. Then we used this approach to investigate the binding conformations of UEV domain proteins with three cyclic peptides inhibitors. We reported the possible cyclic peptide-UEV domain protein binding conformations via 2D PMF free energy profiles and SIE free energy calculations. The residues Trp145, Tyr147, and Trp148 of the native cyclic peptide (CP1) indeed play essential roles in the cyclic peptides-UEV domain proteins interactions. Our findings might increase the accuracy of cyclic peptide-protein conformational prediction, which may facilitate cyclic peptide inhibitor design. Our approach is expected to further aid in addressing the challenges in cyclic peptide inhibitor design. N-terminal ubiquitin E2 variant (UEV) domain of Tsg101 proteins can bind to the PT/SAP motifs on HIV Gag p6 region or Ebola Vp40 proteins [Human Tsg101 proteins are important receptor proteins required for budding of the enveloped viruses ,4; thus,proteins ,8, whereEsomeprazole and tenatoprazole are reported that the two small molecules can inhibit the p6/UEV interactions may be applicable in HIV and Ebola virus infection therap . Howeverenzymatic degradation within minutes. From the structural and pharmacological standpoint, cyclic peptides show higher life time and better biological activity compared with linear form peptides [A linear peptide with a low affinity of 54 uM, the wild-type synthetic peptide (amino sequence: PEPTAPPEE), was reported as an inhibitor of UEV for HIV and Ebola virus infection therapy . Howeverpeptides . Cyclic peptides was repoPeptide\u2013protein docking remains a great challenge for most computational docking programs because of the peptide flexibility ,14. SmalStep 1: Building and optimizing the cyclic peptides by using the EVGAZZ software.Step 2: Docking the cyclic peptides to UEV domain protein receptor by using ClusPro software.Step 3: Selecting possible cyclic peptide-UEV domain protein receptor complex structures from step 2.Step 4: Performing energy minimizations, NTV (1ns), and NPT (1ns) equilibrium simulations by using pmemd.cuda (Amber18).Step 5: Performing 20-ns GaMD equilibrium simulations by using pmemd.cuda (Amber18).Step 6: Performing 300-ns GaMD production simulation four times by using pmemd.cuda (Amber18).Step 7: Performing analysis of reaction coordinates by using AmberTools cpptraj software to obtain 2D potential of mean force profiles (2D PMF) with PyReweighting toolkit.Step 8: The complex structures with the lowest potential of mean force (PMF) values were performed in 10-ns cMD simulations for SIE free energy calculations and binding mode analysis .We followed Miao et al.\u2019s approach, and the ClusPro docking, normal MD, GaMD, and binding free energy calculations were performed in the cyclic peptide-UEV domain protein receptor calculations . This stThe three cyclic peptides were buiThe ClusPro Dock web-server were applied in the cyclic peptide-UEV domain receptors docking simulations ,23. Next3). These initial structures were chosen as refence structures for analyzing the C.M. distances and backbone RMSDs in our PMF profiles. These initial cyclic peptide-UEV domain protein structures were then simulated using the AMBER 18 software with the all-hydrogen amino acid AMBER FF99SB force field [From the ClusPro peptide docking simulations, the complex structures with the lowest potential of mean force (PMF) values were selected and then inserted into TIP3P water molecules box is shown asIf V1 < V2, V1* < V2*. By replacing V* with Equation (2), the relationship is expressed asIf V1 > V2. By replacing V* with Equation (2), the relationship is expressed asCombing Equations (3) and (4) is expressed asFrom Equation (5), we can deriveK0 is the magnitude of the applied boost potential. Standard deviation (SD) of \u0394V must be sufficiently small to ensure accurate reweighting .(8)\u03c3\u0394V=If E = Vmax, we can derive Equation (5) and the result is expressed asAccording to Equations (6) and (7), K0 can be defined asIf E = Vmin + 1/k, we can derive Equation (8) and the result is expressed asThe GaMD boost potential (\u0394V) is given as follows:For our GaMD simulations, the magnitude K0 = 1.0, Vmax and Vmin were obtained through normal MD (cMD) simulations. cE and vdwE are the intermolecular electrostatic and van der Waals interaction energies, respectively. The binding free energies of the UEV domain protein with the three cyclic peptide inhibitors were calculated for 10 ns cMD simulations. The SIE functionequation ,34. \u0394MSAequation . Additioequation .These cyclic peptides were docked into the UEV domain protein structure. The initial cyclic peptide-UEV domain proteins complex structures were selected after sorting the weighted score values. Our cyclic peptide docking results were shown in GaMD simulations has been prove to refine the cyclic peptide-protein receptor complex structures. The C.M. distances and backbone RMSDs of the four times 300 ns-GaMD simulation trajectories were analyzed using AmberTools cpptraj software. The GaMD simulations were reweighted using the PyReweighting toolkitto calculate the 2D PMF profiles Because The predicting and experimental binding free energies (\u0394G) of the three cyclic peptides with UEV domain protein are presented in Crystallographic structures revealed the binding modes of the UEV domain proteins with the native linear peptide, and this binding mode is shown in challenge for most computational docking programs because of the peptide flexibility. Even though these docking methods are fast, these methods still lack thermodynamic refinements and cannot provide accurate peptide binding conformational predictions. These peptide docking methods can\u2019t perform docking simulations with cyclic peptides and can only perform related simulations with linear peptides [Our validation of the Tsg101 UEV protein and the linear peptide (amino sequence: PEPTAPPEE and PDB ID: 3obu) is reasonable and shown in the supporting information. We are confident about our predictions. Cyclic peptides are a yet underexploited candidates for drug discovery. Compared to their linear counterparts, cyclic peptides have enzymatic resistance and lower 3D conformational freedom, which allow them to bind to protein receptors with higher binding affinity and specificity . For spepeptides ,38,39. Tpeptides . Comparepeptides . TherefoGaMD refinement simulations revealed the 2D PMF profiles , and we Since the single point residue mutations of peptides might have different 3D conformations and affect the binding affinities with protein receptors ,43. The For speeding up the cyclic peptides drug discovery, an approach, which combins global docking, normal MD simulations, GaMD simulations, 2D free energy profiles (2D PMFs), and SIE free energy techniques, was applied in studying the interactions between the three cyclic peptides and UEV domain proteins. From the GaMD refinement simulations, our 2D PMF profiles found out the most possible cyclic peptide-UEV domain protein complex structures. Then our SIE binding free energy predictions were close to the experimental binding free energies, and these results confirmed that our cyclic peptide binding conformational predictions seemed reasonable and possible. Next, we analyzed the binding modes of the cyclic peptides with UEV domain protein from 10-ns cMD simulations each cyclic peptide. The hot spot residues Trp145, Tyr147, and Trp148 of the three cyclic peptides can influence UEV domain protein binding affinities. Our study not only can increase the accuracy of cyclic peptide\u2013protein 3D conformational prediction, but this study also can help future cyclic peptide drug discovery. Advancements in computational methods and computing power are expected to further aid in addressing the challenges in cyclic peptide drug design."} +{"text": "NURF binds to histone modifications that decorate the Drosophila polytene male X chromosome and is required to maintain correct organisation of this chromosome. NURF mutants exhibit distorted and decondensed polytene male X chromosomes dependent on the presence of the male-specific lethal (MSL) complex. Here we tested whether mitotic chromosomes similarly require NURF to maintain correct morphology. Surprisingly, although the MSL complex remains associated with mitotic male X chromosomes, NURF is not required to maintain morphology. While the ISWI subunit of NURF is known to remain associated with mitotic chromosomes we show that the NURF specificity subunit Nurf301/BPTF dissociates from chromatin during both Drosophila and human mitosis, further illuminating that NURF is dispensable for mitotic chromosome organisation.The nucleosome remodelling factor (NURF) is an ISWI-class ATP-dependent chromatin remodeling enzyme required both for gene expression and higher order chromatin organisation Drosophila NURF is an ISWI-containing chromatin remodeling complex that catalyzes ATP-dependent nucleosome sliding. By sliding nucleosomes, NURF can alter chromatin dynamics to control both transcription and genome organisation. NURF is composed of four subunits including the catalytic subunit Iswi and a large, highly conserved, NURF-specific scaffold subunit. In Drosophila this large subunit is Nurf301 ) or CG32346, FlyBase ID:FBgn0000541) in humans bromodomain and PHD finger transcription factor (BPTF)) . Nurf301/BPTF contains domains that have the potential to recognize specific histone modifications including a C-terminal bromodomain that recognizes histone H4 acetylated at lysine position 16 (H4K16Ac) . H4K16Ac is enriched on the Drosophila male X chromosome as a result of the actions of the male-specific lethal (MSL) complex and in addition to being bound by Nurf301 can influence the activity of the Iswi subunit of NURF .et al., 1995) that display features of interphase chromosome organisation including topologically associated domains . Previous work has shown that the NURF mutant male polytene X chromosome phenotype is dependent on the presence of the MSL complex and that NURF functions on the male polytene X chromosome by antagonising the MSL complex .Mutations in NURF subunits exhibit striking disruption of male polytene X chromosomes. This includes a characteristic decondensation or \u201cbloating\u201d of male polytene X chromosomes, showing that NURF functions to organise extensive regions of the male polytene X chromosome. Polytene chromosomes are the large chromosomes of the giant nuclei of salivary glands (reviewed in Zhimulev and Koryakov (2009)) that result from as many as ten rounds of endoreplication, where DNA is replicated without intervening mitosis . The large number (1024C) of resultant sister chromatids and homologous chromosomes are held together generating single large chromosomes (Urata Drosophila larvae from both control larvae (1118w) and Nurf301 or Iswi mutant larvae. Mitotic chromosomes were prepared from both male and female larvae. As shown in Fig. 1A no differences in male X chromosome morphology relative to wild-type were detected in either male Nurf301 or Iswi mutant animals. As control the characteristic distorted polytene male X chromosome phenotype could be observed in salivary glands from the same mutant animals used for preparation of neuroblast squashes .Here we investigated whether this requirement is a general feature of all chromosome states. In particular we sought to establish whether NURF is also required for organisation of mitotic male X chromosomes. We examined morphology of mitotic chromosomes in neuroblasts of third instar et al., 2007; Corona et al., 2002). However, immunofluorescence microscopy of mitotic chromosomes using anti-Msl-2 antibodies detected continued presence of Msl-2 on mitotic chromosomes . Thus, even though components of the MSL complex remain associated with mitotic chromosomes, NURF is dispensable for correct mitotic male X chromosome morphology.The lack of a mitotic male X chromosome requirement for NURF could be simply due to the dissociation of the MSL complex from mitotic chromosomes. It was shown previously that the bloated NURF mutant male polytene X chromosome is dependent on the presence of the MSL complex. MSL complex mutants or deletion of high affinity MSL complex binding sites are able to suppress the NURF mutant male polytene X chromosome phenotype . In addition, although mammalian ISWI complexes have been implicated in cohesin loading and maintenance of chromatid cohesion , our analysis showed no discernible defects in sister chromatid cohesion in either male or female Iswi mutants . The lack of observable mitotic chromosome phenotypes in NURF mutants was interesting given previous analysis showing that Iswi remains associated with mitotic chromosomes . However Iswi is the catalytic subunit of at least seven remodeling complexes . To establish whether NURF remains associated with mitotic chromatin we therefore examined chromosome association of the NURF-specific Nurf301 subunit in larval neuroblasts.Further inspection of neuroblast squashes indicated no discernible defects in organisation of mitotic autosomes in male or female et al., 2010). We observed that acid treatment caused either the loss or masking of signal from Nurf301 and certain histone modifications and therefore modified chromosome preparation methods to omit acid treatment, deploying instead an acid-free fixation protocol developed for polytene chromosomes . Interphase nuclei and mitotic chromosomes from larval neuroblasts of control third instar Drosophila larvae were double-immunostained with antibodies against Nurf301 as well as antibodies against histone modifications that are enriched on mitotic chromosomes: anti-histone H3 phosphorylated at threonine position 3 (H3T3p) as well as the phosphomethyl mark (H3T3pK4me3). While we could readily detect Nurf301 in interphase neuroblast nuclei, Nurf301 signal was absent from mitotic chromosomes . This was not due to inability to stain chromatin proteins on mitotic chromosomes as both the H3T3pK4me3 and the H3T3p mark were detected.Initial anti-Nurf301 antibody staining of neuroblast squashes prepared using standard methodologies failed to detect Nurf301 at any stage of the cell cycle. However, these protocols employed acetic acid treatment to assist chromosome spreading . During metaphase Nurf301 was delocalized from chromosomes, re-associating with chromatin from anaphase. In a similar manner human BPTF was detected on interphase HeLa cell chromatin but was not localised to chromosomes during mitosis .These data suggest that NURF is ejected from mitotic chromatin in neuroblasts. To confirm whether this is generally observed in other tissues or organisms we performed immunostaining of Nurf301 mutant male polytene X chromosomes are distorted and decondensed, Nurf301 mutant male mitotic chromosomes are unaffected. Strikingly, while targeting of the MSL complex to polytene chromosomes is sufficient to trigger aberrant chromosome morphology in Nurf301mutants ), the MSL complex is unable to do so on when present on mitotic chromosomes. It is tempting to speculate that this reflects underlying differences in the ability of the MSL complex to promote transcription on polytene versus mitotic chromosomes. Even though recent data indicate that mitotic chromosomes are not transcriptionally silent as has previously been believed, levels of transcription are generally lower .Cumulatively these data indicate that NURF is ejected from mitotic chromatin and is not required for proper mitotic chromosome condensation or cohesion. Moreover, although Drosophila neuroblast mitotic chromosomesPreparation of 2Nurf301 strain and isogenic 1118w control strain are as described). 1Iswi and 2Iswi were obtained from John Tamkun and are as described ). All strains were raised at 22\u00baC. Mitotic chromosomes were prepared as described by Pimpinelli et al. (2010). In brief 3rd instar larval brains were dissected in saline (0.7% NaCl), washed briefly in saline and transferred to hypotonic solution (0.5% Sodium Citrate) for 10 minutes. Brains were cleared in 45% acetic acid on a siliconized coverslip, fragmented using a tungsten needle and squashed onto slides between blotting paper. Slides were frozen in liquid nitrogen, coverslips removed and slides mounted in Vectashield with DAPI (Vector Laboratories). For anti-Msl-2 staining, the same protocol was followed except after freezing samples were fixed in 1% paraformaldehyde for 10 minutes prior to two washes in PBTw buffer . Primary antibody incubation was performed in blocking buffer containing anti-Msl-2 antibodies (1:1000) overnight. Slides were washed three times in PBTw buffer, followed by secondary antibody incubation in PBTw containing Cy3-conjugated anti-Rabbit IgG (H+L) for 2 hours. Slides were washed in PBTw as above, mounted in Vectashield with DAPI (Vector Laboratories) and imaged using a Zeiss LSM780 confocal microscope.The et al. (2006). Briefly, brains were fixed in Brower\u2019s fixation buffer containing 2% formaldehyde ) for 3 minutes, washed in PBT for 2.5 minutes, transferred to 50% Glycerol for 5 minutes, fragmented and squashed onto Poly-L-lysine coated slides. Slides were frozen in liquid nitrogen then processed for double-immunostaining. Slides were incubated with anti-Nurf301 antibodies (1:200 in blocking buffer) overnight at 4\u00b0C then washed with PBTw three times for 10 minutes. Secondary antibody incubation was conducted using Cy3-conjugated AffiniPure Fab Fragment Goat Anti-Rabbit IgG in PBTw for two hours at room temperature, slides were then washed with PBTw three times for 10 minutes and fixed in 1% formaldehyde for 10 minutes. Samples were washed in PBTw as above and incubated with unconjugated AffiniPure Fab Fragment Goat Anti-Rabbit IgG in PBTw for two hours at room temperature to block the first rabbit IgG. Samples were washed with PBTw as above and the second primary antibody reaction was performed in blocking buffer overnight at 4\u00b0C. Rabbit anti-phospho-trimethyl Histone H3 (Thr3/Lys4 & Thr22/Lys23) antibody or rabbit anti-phospho-Histone H3 (Thr3) antibody were used at 1:1000 and 1:1500 respectively. Slides were washed as above and the second secondary antibody reaction performed using FITC-conjugated AffiniPure Fab Fragment Goat Anti-Rabbit IgG for two hours at room temperature in the dark. Samples were washed as above, then mounted and imaged by confocal microscopy as described above.Immunostaining of mitotic chromosomes for Nurf301 and histone modifications followed the same procedure with the exception that after hypotonic treatment brains were processed using an acid-free protocol as described by DiMario Drosophila embryosFixation and permeabilization of Drosophila embryos were collected on apple agar collection plates and rinsed with water onto a Nitex mesh sieve to remove residual yeast. Embryos were dechorionated in 5% sodium hypochlorite solution for three minutes. The embryos were transferred to a 1.5ml Eppendorf tube containing 50% n-heptane (Sigma) and 50% PEM-formaldehyde solution (0.1 M PIPES (pH 6.95), 2 mM EGTA, 1 mM MgSO4, 4% formaldehyde (Pierce)). Embryos were fixed for 25 minutes at room temperature on a rotator. The aqueous layer was removed, 1 volume of methanol was added, and the tube was vortexed for one minute to remove the vitelline membrane. The heptane/methanol phases were allowed to separate and upper phases were removed. Two washes were performed with 1 ml methanol and then with 1:1 dilution of methanol and PBTw (0.1% Tween 20 in 1XPBS). Embryos were washed with 1ml PBTw three times for five minutes on a rotator and immunostaining performed as described above.Huma cell culture and immunofluorescence microscopy2 and cultured in Dulbecco\u2019s Modified Eagle\u2019s Medium (DMEM) containing 10% FBS, 100U Penicillin-Streptomycin (Sigma), 1mM Glutamine (GIBCO) and 1x non-essential amino acids (Sigma). Slides were prepared for immunofluorescence microscopy as described in Kwon et al. (2020), blocked overnight at 4\u00baC in blocking buffer . Primary antibody incubation using anti-BPTF/FALZ antibodies (1:200) was performed in blocking buffer for 2 hours followed by three washes in PBTw buffer . Secondary antibody incubation was performed in PBTw at room temperature for 1 hours using Cy3-conjugated AffiniPure goat anti-rabbit antibody (Jackson ImmunoResearch Cat. 111-167-003) at 1:400 dilution. Slides were washed as above, mounted using Vectashield with DAPI (Vector Laboratories) and imaged using a Zeiss LSM780 confocal microscope.HeLa cells were obtained from the ATTC and STR profile verified . HeLa Cells were incubated at 37\u00baC with 5% CO"} +{"text": "Decisions about asymptomatic COVID-19 patients are always critical, either during initial screening or during recovery. Spread of infection will be inevitable if those patients were left non-isolated. This study aimed not only to survey spectrum of HRCT findings of COVID-19 among asymptomatic and recovered patients but also to record unexpected results and document their impact upon the clinical decision.P < 0.0001); PCR had to be repeated with home isolation (43.3%). Infected health care providers had to stop their duty immediately (20%). Isolated hospitalization replaced routine ward admission (25%). Cautious surgical interference was performed using full personal protective equipment (PPE) (8.3%). Among asymptomatic recovered COVID-19 patients, unexpected large lesions (> 3\u2009cm) were found (70%). Near 50% of lung volume was persistently affected (10%). Secondary fibrosis was striking (33%). Encysted hydro-pneumothorax persisted for a whole month (1.7%). \u201cNo-isolation\u201d decision remained unchanged because of clinical and laboratory stability; however, steroids were prescribed to speed lung recovery.The study was retrospectively conducted, during June and July 2020, on 120 patients proved with COVID-19, during initial HRCT screening or delayed following announcement of recovery. All patients were completely asymptomatic. They included 72 males and 48 females (60%:40%). Their age ranged from 10 to 58\u2009years (mean 35.95 \u00b1 12.25 SD). HRCT was analyzed by three expert consultant radiologists in consensus. Among asymptomatic initially screened COVID-19 patients, additional to GGOs, bilateral consolidative changes were unexpectedly found together with secondary fibrosis (23.3% and 10%). HRCT results significantly impacted the clinical decision (HRCT findings among asymptomatic and recovered COVID-19 patients can be unexpected and can definitely impact the clinical decision. COVID-19 is a highly infectious disease that started in Wuhan in December 2019 and rapidly spread all over the world to be announced as a pandemic by the WHO in March 2020. Infection can be transmitted through respiratory air droplets or via direct contact with contaminated surfaces .While COVID-19 is mainly manifested by fever, cough, dyspnea, and chest tightness, it was reported that around 1-5% of patients can be asymptomatic. Those asymptomatic patients are recommended to be isolated to avoid further spread of infection . PatientDecisions about asymptomatic and recovered patients are always critical. They need to be highly accurate and also rapid because infection spread will be inevitable if those patients were left non-isolated. Considering the PCR low sensitivity, time consumption, high cost as well as non-availability in some countries, HRCT screening has expanded to involve not only those persons who had contact with proved COVID-19 patients but also every patient who will be admitted to a health facility to receive any kind of medical care. This study aims to record unexpected HRCT findings among asymptomatic and recovered COVID-19 patients, also to evaluate their impact upon clinical decision.This study was retrospectively conducted, during June and July 2020, on 120 patients proved with COVID-19; 60 patients were discovered during initial HRCT screening while other 60 patients had delayed abnormal CT findings for 2 to 4\u2009weeks following announcement of recovery. All patients were completely asymptomatic. They included 72 males and 48 females (60%:40%). Their age ranged from 10 to 58\u2009years (mean age was 35.95 \u00b1 12.25 SD).The study was approved by The Ethics Committee of our University Hospital. Patient consent was waived by the Research Ethics Board with assurance of respect of confidentiality of the patients and medical records.Inclusion criteria were completely asymptomatic patients with positive HRCT results during one of the following scenarios: (1) Initial HRCT screening for asymptomatic patients with recent contact history with proved COVID-19 patients or asymptomatic patient with other irrelevant medical condition necessitating hospital admission (both proved positive for COVID-19 using PCR swab tests earlier or later). (2) Follow-up CT scans for asymptomatic recovered COVID-19 patients, performed 2-4\u2009weeks after two consecutive negative PCR results announcing patient recovery.Exclusion criteria were (1) degraded CT scans quality because of respiratory motion artifacts. (2) Any chest symptom relevant to COVID-19.Evaluation of the impact of HRCT results on the clinical decision was done by a single consultant pulmonologist who has 19\u2009years\u2019 experience in the field of infectious lung diseases. Additional correlation with the oxygen saturation and laboratory tests among the recovered patients was also performed.Two MDCT machines were used: Siemens SOMATOM Sensation 64 (Germany) and Toshiba Aquilion CXL/CX 128 (USA). The following CT scanning parameters were used: 1\u2009mm slice thickness, 1\u2009mm detector collimation, 0.6-0.9\u2009s tube rotation, helical mode volumetric HRCT with 100-120 kVp and 80-200\u2009mA, according to the weight of patients and clinical indication. Intra-venous contrast administration was not used.CT scans were evaluated in consensus by three consultant radiologists who were informed with the clinical data and have 15, 19, and 25\u2009years of experience in chest imaging. Multi-planner reconstruction (MPR) was used for image analysis. Post-processing maximum intensity projection (MIP) and minimum intensity projection (Min-IP) reconstructions were performed. Each CT scan was evaluated according to (1) site of lung involvement, (2) universally agreed CT findings with COVID-19 including ground-glass opacities (GGO) with or without consolidative changes in addition to special signs such as \u201cAtoll sign\u201d and \u201cCrazy paving pattern\u201d , manifesP value < 0.05 was considered statistically significant. Online calculators were used (https://www.socscistatistics.com/).The prevalence rate of HRCT findings was estimated as the percentage of patients showing any criterion. Data were compared using a chi-square test and Wide spectrum of HRCT findings were found in this study among both asymptomatic initially screened patients and asymptomatic recovered patients. They included unexpected results regarding the number, the size of the lesions, the site and extension of lung involvement, and the different morphological CT features. All HRCT findings are detailed in Table Timing of positive PCR results was variable; only 31/60 patients (51.7%) had positive PCR swab results at the first trial while 17 patients (28.3%) had positive results at the second trial and the remaining 12 patients (20%) had late positive results at the third trial.Forty-two patients had their initial PCR swab tests before HRCT screening; 26 patients among them had negative PCR results, and then proved to be positive later (2-4\u2009days) after HRCT screening. Other three patients, who were admitted for non-pulmonary medical conditions and performed HRCT screening, showed negative first PCR results then proved to be positive later.All patients were persistently asymptomatic till the PCR proof of COVID-19 infection.Bilateral lung involvement was predominant in 42/60 (70% of patients), the number of lesions exceeded three in 36/60 (60% of patients) and the size of lesions exceeded 3\u2009cm in 18/60 (30% of patients) . Atoll sign that denotes organization process of the disease and even secondary fibrosing changes were also found (each in 10% of patients) who had negative PCR swab tests\u2019 results before HRCT. Repeating PCR tests was requested for them and PCR proved positive 2-4\u2009days later. Among those 42 patients, 12 patients were health care providers and had to stop their duty immediately. Home isolation or isolated hospitalization instead of routine ward admission was the modified clinical decision for three and 15 patients respectively Fig. .Fig. 3ACautious surgical interference using full personal protective equipment (PPE) was carried out in five urgent surgeries instead of routine infection control measures. Delaying a scheduled chemotherapy cycle was decided for two patients. Delaying an elective interventional procedure was also decided for one patient. Strict infection control measures were requested for the radiology unit, which was visited by nine COVID-19 patients Fig. .Fig. 4AExpected simple GGOs were found only in 12/60 (20% of patients) while 40% of patients showed unexpected persistent curvilinear fibro-consolidative changes and also fibrosis on top of GGOs was found in 20/60 (33.3% of patients) Fig. . One patSmall-sized residual lesions (less than 3\u2009cm) were only found in 10/60 (16.7% of patients) while unexpected large lesions (> 3\u2009cm) were found in 70% of patients. Moreover, near 50% of lung involvement was found in 10% of patients , even those patients with secondary fibrotic changes (95-96% O2 Sat/RA). Consequently, the clinical decision remained unchanged with no need for re-isolation or re-hospitalization whatever the prolonged and even unexpected residual CT findings. However, steroids were prescribed for all patients. Also, long term follow-up was recommended to those patients who had persistent secondary fibrosing changes to evaluate its effect on lung function and rule out the possibility for developing secondary interstitial lung fibrosis.All recovered patients in this study had unremarkable laboratory tests and normal range of OThis study surveyed the different HRCT findings among the asymptomatic and recovered patients proved for COVID-19, also recorded the unexpected results, and documented their impact on the clinical decision.Bilateral lung involvement was predominant in this study (70%) conversely to Meng H et al. who founP value < 0.0001) especially among those 29 patients who had initial prior negative PCR swab tests. Modified or new clinical decisions have been made as previously detailed in Table These positive HRCT findings in asymptomatic initially screened patients had a significant impact on the clinical decision as statistically proved (Based on the variable timing of positive PCR results among current asymptomatic initially screened patients, this study agreed with Ai T et al. that CT During COVID-19 disease recovery and healing process, the consolidative changes in the current study were less than that found in Pan F et al. (40% comAgreeing with Sun R et al. who stat2 saturation levels that were observed among these recovered patients. Steroids were prescribed for these patients to speed lung recovery. Long-term follow-up was recommended also to those patients with persistent secondary fibrosing changes to rule out permanent impact on lung functions or possibility for developing secondary interstitial lung fibrosis.These positive HRCT findings in asymptomatic recovered patients did not change the current clinical decision whatever their grade or delayed persistence. No need for re-isolation or re-hospitalization was kept as the same decision in all recovered patients. This was explained by the fact of \u201cradiological lag,\u201d which was previously stated for pneumonia recovery by Bruns AH et al. and descThis study has the advantage for tracing the impact on clinical decision not only surveying CT features but also prolonged follow-up for recovered patient; however, it is limited by the small number of patients and lack of further knowledge about the further clinical outcome for those asymptomatic patients after the end of the isolation period.HRCT findings among asymptomatic and recovered COVID-19 patients can be unexpected and can definitely impact the clinical decision."} +{"text": "MetAP2 is a 67kDa protein which sits at the translation initiation complex and cleaves N-terminal methionine off of nascent peptides. Inhibitors of MetAP2 cause profound weight loss secondary to decreased food intake. These inhibitors also significantly extend longevity in mice in late-life intervention. However, the exact mechanism of action causing decreased food intake is not known. Here we investigated the molecular mechanism and target tissue of a MetAP2 inhibitor\u2019s (Zgn1062) anorectic effects. First we identified the target tissue by testing targeted Zgn1062 delivery to specific brain regions. Delivery to the medio-basal hypothalamus did not have a significant effect but delivery to the lateral ventricle resulted in significantly decrease food intake and body weight after 2 and 14 hours. When we delivered a neuron-targeted AAV encoding MetAP2 shRNA we saw decreased efficacy of MetAP2 confirming the required for neuronal MetAP2 for anorectic effects. To determine the molecular mechanisms we performed RNAseq of wildtype and MetAP2 KO HT1080 cells across a timecourse of Zgn1062 treatment. The main pathway activated across timepoints in MetAP2-dependent manner was P53 signaling. A main P53 target that was upregulated was the known anorectic peptide GDF15. We confirmed GDF15 increases in vivo at both mRNA (liver and intestines) and protein level (serum) in response to Zgn1062. We also found that Zgn1062 treatment reduces senescent cell burden in visceral adipose tissue in vivo and reduces SASP gene expression in fat explants ex vivo. We hypothesize that Zgn1062\u2019s potent P53 activation may play a role in clearance of senescent cells."} +{"text": "P\u2009=\u20090.07), while more significantly pronounced in copy number gains (P\u2009=\u20090.03) in NR compared to ExRs. Delineation of the distribution of CNA burden across the genome identified a greater degree of CNA burden in NR within Chr8 (P\u2009=\u20090.02) and in Chr17 (P\u2009=\u20090.06) and conferred a statistically significant benefit in overall survival. Clinical trial number: NCT01722890\u00a0[ICORG 12/09].Trastuzumab has significantly improved the overall survival of patients with HER2+ metastatic breast cancer (MBC). However, outcomes can vary, with patients progressing within 1 year of treatment or exceptional cases of complete response to trastuzumab for \u226510 years. Identification of the underlying genomic aberrations of \u201cexceptional responders (ExRs)\u201d compared to \u201crapid non-responders (NRs)\u201d increases our understanding of the mechanisms involved in MBC progression and identification of biomarkers of trastuzumab response and resistance. Whole-exome sequencing was performed on six ExRs compared to five NR. The overall fraction of genome copy number alteration (CNA) burden was higher in NR patients ( However, to date only clinical and molecular analysis of this \u201cexceptional\u201d cohort exists. Somatic copy number alterations (CNAs) alter a significant proportion of the cancerous genome. CNAs dominate the breast cancer genome, with somatic mutations in breast cancer genes at low frequencies and mainly characterised in driver genes by high-throughput targeted mutation profiling.5 However, somatic single-nucleotide variants (SNVs) and insertions/deletions in driver genes do contribute to tumour biology.6 It remains unclear as to the extent in which genomic CNA burden can act as a prognostic measure of predicting response to trastuzumab in long-term, never relapse exceptional responders (ExRs) compared to rapid non-responders (NRs). To investigate this hypothesis, we present the first study of whole-exome sequencing (WES) analysing the genome CNA burden of six ExRs compared with five NRs with HER2+ MBC patients treated with trastuzumab.Historically, HER2+ metastatic breast cancer (MBC) was designated as an incurable disease. The introduction of anti-HER2 therapies such as trastuzumab has markedly improved survival7 and duplicate reads were marked. Base recalibration was conducted with GATK.8 Variant calling was performed using Mutect2 from GATK (4.1.3). CNAs were identified by EXCAVATOR2.9 Variant statistics and tumour mutational burden (TMB) were calculated by maftools.12A retrospective, single institution review from 2000 to 2018 identified 295 HER2+ MBC who received treatment with trastuzumab in the metastatic setting. Informed consent was obtained and approved by St. Vincent\u2019s University Hospital Ethics Committee. HER2+ immunohistochemical status was confirmed by pathologist (C.Q.). Analysis of patients with never relapse and >3-year overall survival follow-up data identified 40 patients. A further refined analysis revealed 11 patients with a minimum overall survival of 10 years (range 10\u201319 years). We performed WES on 6/11 of these patients and 5 corresponding NRs \u226414 months) in NRs compared to ExRs and in chromosome 17 ; however, it does stratify patients and NR (n\u2009=\u20094) cases. Of our small cohort, we observed a higher median of SNV in ExRs (median\u2009=\u20091621) compared to NRs (median\u2009=\u2009638); albeit this large variance was mostly driven by an unusual SNV in ExR patient 1 , HSPG2 (60% of ExR and 75% NR samples), SYNE1 (40% of ExR and 75% NR samples), SYNE2 (80% of ExR and 50% NR samples) and MACF1 (40% of ExR and 75% NR samples) therapeutic response in the metastatic setting.Identification of genomic alterations associated with exceptional response and survival may improve risk assessment and treatment strategies for HER2+ MBCs. This is the first study to propose that CNA burden in HER2+ MBC ExRs may represent a novel prognostic predictor to trastuzumab response. Despite the limited sample size, we observed a trend in which the overall CNA burden was lower in ExRs compared to NRs; moreover, individual CNA analysis per chromosome revealed that specific chromosomes 8 and 17 were more altered in NR genomes compared to ExRs, and stratified analysis revealed significantly poorer overall survival. CNA burden was previously shown to be associated with overall survival and disease-specific survival in breast cancer, with chromosome 8 along with chromosomes 1 and 16 carrying the highest CNA burden, suggesting a further role of chromosome 8 in prognosis.Supplementary files"} +{"text": "The COVID 19 pandemic caused from SARS CoV 2 was declared on March 11, 2020 . HydrogeThe use of HI also against virus infection is well documented in the scientific literature. Goyal et al. studied the effectiveness of a vapour of HI versus different viruses: feline calicivirus; human adenovirus type 1; transmissible gastroenteritis coronavirus of pigs (very similar to SARS-CoV); avian influenza virus (AIV); and swine influenza virus (SwIV). Authors demonstrated that adenovirus, TGEV and AIV died at the lowest vaporized HI volume tested 25 ml). They evaluated that the total exposure time, including injection and aeration, was 2-3 hours, varying with the amount of hydrogen peroxide being vaporized ml. They. RudnickAn hydrogen peroxide spray for decontamination of surfaces in hospitals was successfully tested by Cadnum et al. They observed that a 1.4% solution of hydrogen peroxide (IHP) spray disinfectant could be very effective against Staphilococcus aureus and strains of vancomycin-resistant enterococci . In a veThe authors declare no competing interests."} +{"text": "Oryza sativa respiratory burst oxidase homologue B (OsRBOHB) during compatible and incompatible interactions between rice epidermal cells and the pathogenic fungus Pyricularia oryzae (syn. Magnaporthe oryzae). We characterized the functional role of ROS focal accumulation around invading hyphae during P. oryzae infection process using the OsRBOHB inhibitor diphenyleneiodonium (DPI) and the actin filament polymerization inhibitor cytochalasin (Cyt) A. OsRBOHB was strongly induced during incompatible rice\u2013P. oryzae interactions, and newly synthesized OsRBOHB was focally distributed at infection sites. High concentrations of ROS focally accumulated at the infection sites and suppressed effector biotrophy-associated secreted (BAS) proteins BAS4 expression and invasive hyphal growth. DPI and Cyt A abolished ROS focal accumulation and restored P. oryzae effector BAS4 expression. These results suggest that ROS focal accumulation is able to function as an effective immune mechanism that blocks some effectors including BAS4-expression during P. oryzae infection. Disruption of ROS focal accumulation around invading hyphae enables successful P. oryzae colonization of rice cells and disease development.The reactive oxygen species (ROS) burst is the most common plant immunity mechanism to prevent pathogen infection, although the exact role of ROS in plant immunity has not been fully elucidated. We investigated the expression and translocation of Plant pathogen effectors facilitate parasitism by suppressing pathogen-associated molecular pattern (PAMP) perception or modifying plant physiology to support pathogen growth and colonization ,2,3. WhiOryza sativa respiratory burst oxidase homologue (OsRBOH) proteins in the rice genome, and OsRBOH genes were differentially expressed depending on the rice tissue and environmental stresses , incubated for 5 min in the dark, and ROS was detected by luminometer under dark condition.ROS was quantified by performing a chemiluminescence assay . Rice shn = 4) inoculated with different P. oryzae spore suspensions were sampled at different time points . RNA extraction was performed using TRIzol reagent according to the manufacturer\u2019s protocol. Then, 2 \u00b5g of RNA was used to synthesize first-strand cDNA in 20 \u00b5L total reaction mixture using cDNA synthesis kit according to the manufacturer\u2019s protocol. OsRBOHB expression was determined using 1 \u00b5g cDNA and Real-Time Quantitative Reverse Transcription Polymerase Chain Reaction . The qRT-PCR assay was performed using TOPreal qPCR 2 \u00d7 PreMIX according to the manufacturer\u2019s protocol. The Ubiquitin (LOC_Os06g46770) transcript level was used to normalize the OsRbohB transcript level. Relative gene expression level was determined by comparing with the mock sample at 0 hpi. The primers used in this study are listed in Leaf sheaths of rice cv. HY (n) is given in the Figure legends. Statistical analyses were performed using Prism 7 software .All experiments were repeated independently more than three times. Each biological replicate contained independent leaf sheath samples taken from different rice plants. The number of biological replicates ("} +{"text": "Mitochondrial dysfunction causes many poorly understood diseases, such as Leigh Syndrome, that are often caused by dysfunctions in proteins involved in the electron transport chain. My lab previously reported mTOR is pathologically involved in the neurodegenerative phenotype and premature death of mice missing the Complex I subunit Ndufs4 (Ndufs4-/- mice). We discovered treatment with rapamycin extends lifespan, reduces neuroinflammation, and attenuates the neurodegenerative phenotype in these mice, although the mechanisms remain unclear. Rapamycin-treated Ndufs4-/- mice exhibited decreased activation of the mTORC1 pathway. It also deactivated the mTORC2 pathway. We observed that phosphorylation of the canonical protein kinase C (PKC) isoforms decreased more than any other kinases, leading us to hypothesize its deactivation contributes to the observed lifespan extension. To test this, we treated Ndufs4-/- mice with three different PKC inhibitors: the pan-PKC inhibitors GO6983 and GF109203X, and the PKC-\u03b2 specific inhibitor ruboxistaurin. Similar to rapamycin, all three drugs were able to significantly delay the onset of neurological symptoms (i.e. clasping) and increase survival. We also observed that PKC-\u03b2 inhibition reduced skin inflammation to suppress the hair loss phenotype displayed by Ndufs4-/- mice at weaning. We further discovered PKC-\u03b2 inhibition reduces neuroinflammation by deactivating the NF-kB inflammatory pathway. These results suggest that mTORC2 may play a critical role in the etiology of mitochondrial diseases such as Leigh Syndrome."} +{"text": "In older humans, multiple chronic diseases and increased life expectancy impose a disproportionate socioeconomic burden. Dietary interventions are valuable strategies for promoting healthy aging. Caloric restriction (CR) without malnutrition is a robust intervention able to delay disease onset and increase survival in model organisms. However, the impracticability of chronic CR outweighs the potential long-term benefits in humans. Time-restricted feeding (TRF), i.e. the limitation in the timing of food intake without necessarily reducing caloric intake, can protect against metabolic disorders through the synchronization of the circadian rhythm. This study compares whether limiting access to ad libitum (AL) food for a few hours per day mimics the beneficial effects of a CR diet. A large cohort of C57BL/6J female mice (n=250) was distributed into five feeding paradigms at midlife: AL, TRF for 8 hours, TRF for 4 hours, 20% CR and 20% CR fed twice a day (CRx2). Pathological analyses at death reveal a shift in fatal neoplasms toward an older age in TRF8 mice. AL mice had the highest prevalence of tumors (93%) and TRF4 had the lowest (77%). The highest tumor burden was observed in AL mice while CRx2 animals had the lowest number of neoplasms. Histiocytic sarcoma and lymphoma were the most represented malignancies, with CR mice exhibiting the highest rate of histiocytic sarcoma (75%) and the lowest rate of lymphoma (10%). These results indicate that time- and calorie-restricted feeding regimens can slow down malignant neoplasm progression and extend health span in female mice, even when started in adulthood."} +{"text": "SCN9A gene variants, in particular a c.1921A>T p.(Asn641Tyr) substitution, have been identified as a likely autosomal dominant cause of febrile seizures/febrile seizures plus and other monogenic seizure phenotypes indistinguishable from those associated with SCN1A, leading to inclusion of SCN9A on epilepsy gene testing panels. Here we present serendipitous findings of genetic studies that identify the SCN9A c.1921A>T p.(Asn641Tyr) variant at high frequency in the Amish community in the absence of such seizure phenotypes. Together with findings in UK Biobank these data refute an association of SCN9A with epilepsy, which has important clinical diagnostic implications.Many studies have demonstrated the clinical utility and importance of epilepsy gene panel testing to confirm the specific aetiology of disease, enable appropriate therapeutic interventions, and inform accurate family counselling. Previously, SCN9A gene and forms of epilepsy, leading to inclusion of the SCN9A gene in diagnostic testing panels. In this paper we present the results of our genetic findings and follow-up studies of SCN9A gene alterations in the Amish community and UK Biobank, alongside a fresh appraisal of previous studies investigating the role of SCN9A in epilepsy. Together our findings strongly refute an association between SCN9A and epilepsy. This work has important clinical implications and should lead to the re-evaluation of SCN9A by expert groups and its removal from genetic testing panels, preventing future genetic misdiagnosis which may have devastating and sometimes lethal consequences.Epilepsy is defined as a tendency to have seizures, affecting around 1:100 people worldwide. Some genetic types of epilepsy can be diagnosed using a test that examines genes that have previously been shown to cause epilepsy when affected by genetic alterations. Identifying the genetic cause of a patient\u2019s epilepsy can help determine which treatments are likely to be helpful, provide prognostic information and inform accurate family counselling. The genes that are included in diagnostic epilepsy gene testing panels are decided by reviewing published scientific studies. Previous publications have described an association between the SCN1A-associated seizure disorders where commonly used sodium-channel-blocking medications carbamazepine, vigabatrin and lamotrigine should be avoided because they may worsen the condition by inducing and/or prolonging seizures ) which are present at population allele frequencies inconsistent with them being causative of a monogenic seizure disorder variant burden analysis, of Caucasian epilepsy cases versus controls in UK Biobank exome data, defined no enrichment of plausibly causative rare SCN9A variants Click here for additional data file.S1 TableSCN9A variants examined are more frequent in controls than cases except p., where there is no significant difference between cases and controls (2 sided Fisher\u2019s exact test p = 0.51)Allele frequencies are calculated from Biobank exome data (SPB pipeline), available for 49,959 individuals at the time of publication. Variant frequencies are separated by presence or absence of epilepsy as defined in our methods (above). All (DOCX)Click here for additional data file.S2 Table1 gnomAD v2.1.1 non-neuro cohort.Abbreviations: AC, Allele count; aCGH, array comparative genomic hybridization AF Allele frequency; BECTS, benign partial epilepsy of childhood with centrotemporal spikes; FS, Febrile Seizures; GEFS+, generalised epilepsy with febrile seizures plus; Hom. Homozygous individuals; NGS, Next-generation sequencing; TLE, temporal lobe epilepsy. (DOCX)Click here for additional data file.S3 TableSCN9A variants predicted to have an impact on SCN9A amino acid sequence were compared between cases and controls, with a small but not significant increase observed in controls (1 sided Fisher\u2019s exact test p = 0.398).Allele frequencies of (DOCX)Click here for additional data file.S1 Figwww.gtexportal.org) for nine of the ten VGSC-\u03b1 genes. Unlike the epilepsy-related VGSC-\u03b1 genes , SCN9A is expressed primarily in peripheral nerves.Expression data reproduced from the Genotype-Tissue Expression (GTEx) portal ((DOCX)Click here for additional data file.S2 FigTop of genogram (VGSC-\u03b1 domains) shows the domain structure of VGSC-\u03b1 genes, with four ion transporter Pfam domains (blue boxes) separated by interspersed disordered regions (beige boxes). Beneath, SCN9A is shown alongside the two VGSC-\u03b1 genes that are both associated with epilepsy and have Pfam annotated ion channel domains (SCN1A and SCN3A), with the amino acid position of each domain indicated in each molecule (\u2018positions\u2019 row). The number of ClinVar pathogenic annotated variants (with at least one pathogenic / likely pathogenic annotation) in each is shown below each domain. The four domains with the highest variant density are indicated by a darker shade. For SCN1A (which has a large number of disease-associated variants described) this is also calculated per amino acid to aid interpretation of the mutation load relative to the size of each of domain. Variants in SCN9A are separated into those associated with pain syndromes, and putative variants proposed to be associated with seizures; those associated with both are recorded twice, and those with no phenotypic description have been excluded. This shows that the epilepsy-related VGSC-\u03b1 genes display significant spatial clustering of putative disease-associated variants within regions known to be crucial for molecular function, in particular the ion transporter domain regions (shown by dark blue shading) and the interlinking regions between transporter domains 3 and 4 (shown by dark beige shading). For SCN9A, the expected clustering of variants in functionally important regions is only seen for pain-associated phenotypes with variants proposed to be associated with seizure disorder phenotypes displaying no spatial clustering, consistent with a benign nature.(DOCX)Click here for additional data file."} +{"text": "Dear Editor,TP53 mutated AMLs are frequently resistant to intensive treatments and we recently showed that subclonal TP53 mutations confer an equally devastating prognosis [TP53 mutated subclones display characteristics of leukemic stem cells (LSCs) thus contributing to relapse or resistant disease. We, therefore, tested LSC properties of these subclones in a recently developed, highly sensitive humanized bone marrow (BM) ossicle xenotransplantation mouse model [Despite extensive efforts to develop novel therapies, the prognosis of patients with acute myeloid leukemia (AML) is still poor. One of the reasons is the genetic heterogeneity of AML with the majority of patients exhibiting distinct mutational subclones and diverse biological characteristics , 2. TP53se model , 5.null (NSG) mice. These MSCs underwent endochondral ossification leading to the formation of a humanized BM ossicle microenvironment. We transplanted three diagnostic, T cell depleted AML specimens into ossicle-bearing NSG mice either by tail vein or direct intraossicle injection. Two specimens showed subclonal TP53 mutations with a variant allele frequency (VAF) of <20%, one a clonal TP53 mutation serving as control from healthy donors were expanded in vitro and subcutaneously injected into four sites of immunodeficient NOD/SCID/\u03b3of <0.1% .TP53 mutations, CD33+ leukemic cells constituted the predominant cell population within the graft. Interestingly, differentiation into CD19+ B-lymphoid cells was also observed and the subclonal TP53 mutations could be detected in both, the engrafted myeloid and lymphoid compartments being in line with our previous report on clonal TP53 mutations in AML [TP53 mutations do also face an adverse prognosis since LSCs and pHSCs are considered less vulnerable to cytotoxic therapy ultimately giving rise to relapsed or resistant disease [TP53 aberrations was also shown in murine models exposed to genotoxic stress [All three AML specimens showed engraftment in humanized ossicles as well as mouse BM. However, human cells preferentially engrafted in the humanized ossicle microenvironment Fig. . For botts Figs.\u00a0,3. Theses in AML . Stem ce disease . Similarc stress . FinallyESM 1(PDF 2815\u00a0kb)"} +{"text": "Scientific Reports 10.1038/s41598-020-64931-3, published online 19 May 2020Correction to: The below paragraph contains references to articles hypothesising the same concept which has now been experimentally demonstrated; these were omitted from the Discussion section of the Article.2. Further, the possibility of combining enzymatic reagents for induction of posterior vitreous detachment (PVD), using hyaluronidase for liquefaction and plasmin for dehiscence has been successfully tested in diabetic rats3\u201d.\u201cThe concept that we experimentally demonstrated has been previously theoretically proposed"} +{"text": "Apis mellifera sahariensis (Apidae) belonging to the African lineage. The assembled circular genome has a length of 16,569\u2009bp which comprises 13 protein coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and AT rich region.We present the complete mitochondrial genome of honey bee subspecies, Apis mellifera sahariensis is a subspecies of honeybee (Apis mellifera) belonging to the African lineage and is found in the oases of the Sahara to the south of the Atlas Mountains, A\u00efn-Sefra, Bechar, Algeria and Apis mellifera syriaca (Haddad a Haddad from theApis mellifera sahariensis was sequenced using Illumina HiSeq platform (150\u2009bp paired-end chemistry) at Genotypic Technology Pvt. Ltd. . We assembled a subset of the generated raw reads from the mitochondrial genome and annotated the assembly with the MITOS webserver , T\u2009=\u20097165\u2009bp (43.24%), G\u2009=\u20091595\u2009bp (9.63%), C\u2009=\u2009924\u2009bp (5.58%) with an overall AT-rich percentage of 84.80%. A total of 37 genes which include mitochondrial protein coding genes for oxidative phosphorylation along with 22 tRNA genes as well as genes for the large and small ribosomal RNAs (rrnL and rrnS) were annotated. Homology search of Apis mellifera sahariensis against available Apis mitogenomes resulted in high homology and similar gene arrangement. Phylogenetic analysis showed high similarity of Apis mellifera sahariensis with Apis mellifera intermissa (KM458618) .Apis mellifera sahariensis has been submitted in NCBI GenBank under the accession no. MF351881.The mitochondrial genome sequence of"} +{"text": "Takotsubo cardiomyopathy (TTC) is an increasingly recognized acute cardiac syndrome (ACS), which underlying pathophysiological mechanisms are still controversially discussed. Cardiovascular magnetic resonance imaging (CMR) contributes to an understanding and differential diagnosis of this new entity by demonstrating the absence of irreversible injury (delayed enhancement) but oedema formation on T2-weighted images. However, clinical experience with CMR in this entity is still limited and is based mainly on relatively small case series.The aim of this study was therefore to evaluate CMR criteria for the diagnosis/differential diagnosis of TTC and the assessment of potential pathophysiological mechanisms as well as additional findings by using a comprehensive CMR approach.Between January 2005 and October 2008 81 consecutive patients, showing a left ventricular dysfunction with apical ballooning not explainable by the coronary artery status and initially admitted with ACS underwent CMR using a 1.5 T MRI scanner. Left ventricular function, T2-weighted spin echo sequence for oedema and delayed enhancement (DE) images after administration of Gadobutrol were assessed. Additionally, in the last 20 patients T2-weighted triple-inversion-recovery imaging to calculate the edema ratio (ER) and T1-weighted imaging before and after contrast agent administration to calculate the myocardial global relative enhancement (gRE) were performed for detection of inflammation.CMR revealed diagnosis of myocardial infarction in 18 (22.2%) patients and diagnosis of myocarditis in 9 (10.8%) patients with typical patterns of DE. In all other 54 (67.0%) patients no DE was detected, consistent with the diagnosis of TTC. Of these 20 patients (37%) showed focal oedema, 17 (31%) initial right ventricular involvement and 19 patients a pericardial effusion (PE) (35%). Of the last 20 TTC patients 9 showed elevation of both inflammatory markers (ER and gRE) with concomitant PE indicating acute inflammation. Follow-up CMR after three months showed complete normalization of left ventricular function and inflammatory parameters in the absence of DE, oedema and PE.CMR has incremental value for differential diagnosis, pathophysiological insights and detection of additional findings in TTC. Therefore CMR should be performed in all patients with suspected TTC for further differential diagnosis and guidance of medical therapy. Moreover, our results support the probable underlying cause of inflammation in TTC. If inflammation is the primary cause or secondary phenomena due to sympathetic overdrive needs further research."} +{"text": "Heterotrigona itama is a species of stingless bee recently domesticated (or reared) for honey production in a few Southeast Asian countries namely Malaysia and Indonesia. Being categorized in the clade Corbiculata together with the honeybees (Apis spp.) and bumble bees (Bombus spp.), the stingless bees are highly social in which the colony members are subjected to labor division where a queen functions as the reproductive caste. In this data article, we provide a resource encompassing a transcriptome profile (de novo assembled) from H. itama queen larva \u2013 the first report of transcriptome assembly for this species. The generated data is pivotal for the characterization of important genes and biological pathways in order to further improve our understanding on the developmental biology, behavior, social structure and ecological needs of this eusocial hymenopteran insect from the molecular aspect. The raw RNA sequencing data is available at NCBI Sequence Read Archive (SAR) under the accession number SRP230250 and the assembled reads are deposited at DDBJ/EMBL/Genbank as Transcriptome Shotgun Assembly (TSA) under the accession GIIH00000000. A queen larva at feeding stage (3rd instar) was collected from a mother colony and snap-frozen in liquid nitrogen (N2), and high quality total RNA was extracted before sent for sequencing through paired-end Illumina sequencing technology. De novo assembly was performed using Trinity and contigs were annotated against seven databases using multiple software. An overview of the data and sequencing assembly of H. itama data is presented in The dataset contains raw sequencing data obtained through the transcriptome sequencing of a female 22.1The queen larva, whose species had been validated through DNA barcode (cytochrome oxidase subunit I), was picked from a very healthy colony that had been placed at a dedicated research plot adjacent to a secondary forest for more than 6 months. Larva instar 3 (feeding stage) was selected for RNA sequencing since this stage was reported to be the \u2018deciding stage\u2019 for caste differentiation in female stingless bee larvae . Total R2.2Escherichia coli polymerase I to generate the second strand by nick-translation followed by two rounds of cDNA purification using AMPure XP beads. The cDNA was then proceeded for terminal repair, A-tailing, ligation of sequencing adapters, size selection and PCR enrichment. For library quality assessment, the cDNA library concentration was determined using Qubit 2.0 fluorometer (Life Technologies), and the insert size was checked on Agilent 2100 and quantified to greater accuracy by quantitative PCR (qPCR) (library activity >2 nM). Finally, the prepared cDNA libraries were fed into Illumina machines according to activity and expected data volume.Prior to Complementary DNA (cDNA) libraries preparation and sequencing, quality check (QC) of the total RNA was done as follows: preliminary quantitation (Nanodrop), degradation and contamination tests through agarose gel electrophoresis, and final integrity and quantitation tests (Agilent 2100). The RNA was then processed following these steps: Enrichment of mRNA using oligo(dT) beads, removal of rRNA using a specialized kit and fragmentation of mRNA. Afterward, the cDNA was synthesized from the mRNA fragments using random hexamers and reverse transcriptase. Following the first-strand synthesis, a custom second-strand synthesis buffer (Illumina) is added together with dNTPs, RNase H and 2.3The raw data from Illumina was transformed to Sequence Reads by base calling and recorded in a FASTQ file. Raw reads were cleaned/filtered as follows: (1) removing reads with adaptor contamination, 2) removing reads when uncertain nucleotides constitute more than 10% of either read (N\u00a0>\u00a010%) and 3) removing reads when low quality nucleotides constitute more than 50% of the read. De novo transcriptome reconstruction was carried out using Trinity (version r2014-04-13p1) with a minimum read length of 200 and k-mer\u00a0=\u00a025. The Trinity workflow followed the Inchworm, Chrysalis and Butterfly modules [ removing removingGene functional annotations were carried out using Diamond (v0.8.22), KAAS (r14 0224), NCBI Blast (v2.2.28+), hmmscan (HMMER 3) and blast2go (b2g4 pipe_v2.5) software. A total of seven databases including Nr, Nt, KO, Swiss-Prot, Pfam, GO and KOG"} +{"text": "Experience sampling paradigms provide new opportunity for early identification of mild cognitive impairment (MCI). We investigated two research questions: (1) is time to complete a repeatedly administered survey a reliable and valid measure of cognition? (2) does this measure distinguish MCI status? To answer these questions, we leveraged ecological momentary assessment (EMA) data from the Einstein Aging Study, where older adults (N=240) completed six daily surveys and cognitive assessments on smartphones over 14 days. Q*bert had good between-person reliability after two days (~11 EMAs) and excellent reliability from three to fourteen days. Q*bert moderately correlated with ambulatory cognitive measures of processing speed and memory binding (p\u2019s < .001) and was significantly slower in those with MCI (p < .001). We propose q*bert as a reliable, valid, and unobtrusive measure of cognition when ambulatory cognitive assessments are not feasible."} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-020-63622-3, published online 20 April 2020Correction to: This Article contains errors in the in-text citations. In the Methods section, under the subheading \u2018Climate data and future climate projections\u2019,58).\u201d\u201cTo derive future precipitation and potential evapotranspiration, we used outputs from 16 downscaled global climate models (GCMs) that were part of the Intergovernmental Panel on Climate Change\u2019s (IPCC) Fifth Coupled Model Intercomparison Project, (CMIP5should read:54).\u201d\u201cTo derive future precipitation and potential evapotranspiration, we used outputs from 16 downscaled global climate models (GCMs) that were part of the Intergovernmental Panel on Climate Change\u2019s (IPCC) Fifth Coupled Model Intercomparison Project, :\u201cFuture projected precipitation is an output from the CMIP5 models. Mean annual temperature (Tmean) and annual temperature range (Trange) were the outputs used in conjunction with solar radiation (RA) to model future potential evapotranspiration (PET) using the Hargreaves-Samani equation (Eq.\u00a0(1)57 and therefore was also used for the future projections.\u201dThe Hargreaves-Samani equation was used to calculate the modern AI used in this studyshould read:54,58\u201361):\u201cFuture projected precipitation is an output from the CMIP5 models. Mean annual temperature (Tmean) and annual temperature range (Trange) were the outputs used in conjunction with solar radiation (RA) to model future potential evapotranspiration (PET) using the Hargreaves-Samani equation (Eq.\u00a0(1)57,58 and therefore was also used for the future projections.\u201dThe Hargreaves-Samani equation was used to calculate the modern AI used in this study"} +{"text": "A 44-year old male patient was seen at the emergency department with dyspnea. The electrocardiogram (ECG) showed diffuse repolarization disorders Fig.\u00a0a. A comp. Enhancing myocardial fibres are visible as a subtle haze at the edges of the mass , T1 and T2 imaging showed hyper intensity and balanced Steady-State Free Precession (bSSFP) sequence demonstrated chemical shift artefacts around the mass Below is the link to the electronic supplementary material."} +{"text": "Loneliness is a risk factor for dementia, however it\u2019s relationship with cognitive health during midlife is unclear. We evaluated whether loneliness was associated with profiles of objective and subjective memory in younger and middle-aged adults. Participants (aged 25 to 64 years) underwent an initial loneliness assessment, followed by 14-days of momentary (5 per day) cognitive assessments (objective memory) and daily ratings of memory (subjective memory). Cluster analysis was conducted using person-level means of objective and subjective memory. Three clusters were identified: (1) highest objective and subjective memory (9%); (2) lowest subjective but not objective memory (84%); (3) lowest objective but not subjective memory (7%).There was a trend for higher levels of loneliness in Cluster 2 relative to Clusters 1 and 3. Results suggest that loneliness is more closely related with subjective than objective memory during midlife and are informative for development of interventions targeting cognitive health. Part of a symposium sponsored by the Measurement, Statistics, and Research Design Interest Group."} +{"text": "Luteovirus, family Luteoviridae) from white clover in Germany. The complete genome of the isolate (JKI ID 23556) consists of 5,858 nucleotides and displays 94.98% nucleotide identity to its most similar SbDV relative (GenBank accession number MN412736).In this study, we present the complete genome of a new isolate of soybean dwarf virus (SbDV) from white clover in Germany. The complete genome of the isolate (JKI ID 23556) consists of 5,858 nucleotides and displays 94.98% nucleotide identity to its most similar SbDV relative (GenBank accession number MN412736).In this study, we present the complete genome of a new isolate of soybean dwarf virus (SbDV) (genus Trifolium repens L., family Fabaceae) is one of the most important pasture legumes, covering a wide range of climatic regions in the world (Acyrthosiphon pisum) to faba bean plants , and transmission was confirmed by both ELISA and RT-PCR. SbDV is a single-stranded positive-sense RNA virus and belongs to the genus Luteovirus (family Luteoviridae) . In GermV. faba leaves using the innuPREP RNA minikit , and then rRNA was depleted with the RiboMinus plant kit (Invitrogen) following the manufacturers\u2019 instructions. cDNA was synthesized using random octanucleotide primers and ProtoScript II reverse transcriptase (NEB), and second-strand cDNA synthesis was performed using the NEBNext Ultra II nondirectional RNA second-strand synthesis module kit (NEB) according to the manufacturer\u2019s instructions. The libraries were prepared using the Nextera XT library kit (Illumina) and were sequenced on a MiSeq v3 platform (2\u2009\u00d7\u2009301 bp).To sequence the whole genome of SbDV JKI ID 23556, we followed the steps described previously . BrieflyNC_003056) by mapping with the reference tool; 10,303 reads were mapped, with a mean coverage of 461\u00d7. The GC content of the complete genome of SbDV isolate JKI ID 23556 is 44.2%. A direct BLASTn search of the NCBI database showed that JKI ID 23556 shared the most similar nucleotide identity of 94.98% with SbDV GenBank accession number MN412736. A BLASTp search showed that the SbDV JKI ID 23556 proteins shared between 93.6% and 99.5% amino acid identities with their homologues from SbDVs in the NCBI database were analyzed using Geneious Prime v2020.1.2 software. The reads were quality trimmed and size filtered to >50 nucleotides. The complete sequence of JKI ID 23556 was assembled according to the SbDV reference genome (GenBank accession number database . These rMT543032. The raw data were deposited in the Sequence Read Archive (SRA) under BioProject accession number PRJNA524397, BioSample accession number SAMN15056231, and SRA accession number SRX8424802.The complete genome sequence of SbDV isolate JKI ID 23556 was deposited in GenBank under accession number"} +{"text": "Embryonic stem cells have demonstrated the potential to restore the myocardium. MRI is an ideal method to evaluate myocardial cell therapy. Superparamagnetic iron oxide nanoparticle (SPIO) has been widely used to monitor stem cell therapy. However, this technique does not provide any biologic information of cell viability.2+ is known to be able to enter viable cells through the voltage gated Ca2+ channel and subsequently can shorten T1 relaxation time generating bright signal on T1 weighted sequence.This novel reporter gene (RG) is designed to express antigenic epitopes on the surface of embryonic stem cell (ESC). Employing SPIO-conjugated monoclonal antibodies, MRI signal specific to hESC viability can be generated. Mnc-myc, HA epitopes and firefly luciferase (Luc) on the cell surface. This fusion protein has been designed to be anchored on the cell surface by PDGFR transmembrane domain. Both c-myc and HA epitopes are the molecular targets for MRI viability signal using SPIO conjugated monoclonal antibodies. H9 hESC female line was tranduced with this RG using a p2K7 lentiviral vector. hESC-RG incubated with anti c-myc and HA microbeads were scanned by 3 T MRI using a high array knee coil. For MEMRI, 0.5 mL of 5 mM MnCl2 was injected intraperitoneally after transplanting mESC onto mouse hindlimbs. Mice were scanned using Spin echo sequence by 3 T MRI.MRI RG was constructed driven by EF1\u03b1 promoter to express In vitro MRI showed significant dephasing signal generated from SPIO conjugated antibody for c-myc and HA (Figure In vivo MEMRI could show significant enhancement of viable mESC on the mouse hindlimbs using T1 weighted sequence (Figure Functional expression of hESC-RG was confirmed by FACS, bioluminescence and Prussian blue staining. A Figure . MagnetiA Figure . In vivoe Figure .Figure 1In vivo molecular signal of hESC viability will be feasible using this innovative MRI RG. MEMRI enabled in vivo evaluation of viability of stem cells.The novel MRI RG enabled viable embryonic stem cells to generate significant molecular MRI signal."} +{"text": "Reduced Pilra , Nupr1 , and Kap as well as increased Ccl20, S100a8/a9 , Ifna4, and Ltb4r2 indicated that inflammation-related molecular dysregulation could be a \u201ccommon\u201d feature after IUGR of different origins. Network analyses of transcripts and predicted upstream regulators hinted at proinflammatory adaptions mainly in LIG and dysregulation of AMPK and PPAR signaling in LP pups. The latter may increase susceptibility towards obesity-associated kidney damage. Western blots of the most prominent predicted upstream regulators confirmed significant dysregulation of RICTOR in LP (PND 7) and LIG pups (PND 1), suggesting that mTOR-related processes could further modulate kidney programming in these groups of IUGR pups.This study was performed to identify transcriptional alterations in male intrauterine growth restricted (IUGR) rats during and at the end of nephrogenesis in order to generate hypotheses which molecular mechanisms contribute to adverse kidney programming. IUGR was induced by low protein (LP) diet throughout pregnancy, bilateral uterine vessel ligation (LIG), or intrauterine stress (IUS) by sham operation. Offspring of unimpaired dams served as controls. Significant acute kidney damage was ruled out by negative results for proteins indicative of ER-stress, autophagy, apoptosis, or infiltration with macrophages. Renal gene expression was examined by transcriptome microarrays, demonstrating 53 contains supplementary material, which is available to authorized users. Adverse environmental conditions during intrauterine and early postnatal life have been linked to the predisposition for non-communicable diseases in later life . Low birThe developing kidney is particularly susceptible to adverse environmental impacts since even small changes in gene or protein expression during critical time spans may lead to severely altered renal outcome . In humaVarious experimental models of IUGR have been developed to elucidate the molecular links between insufficient intrauterine conditions and programming of adverse renal outcome. The rat models of low protein (LP) diet throughout pregnancy or bilateral ligation (LIG) of the uterine vessels during terminal pregnancy have been most widely used since they were designed to represent the most common causes of IUGR in humans, namely either malnutrition or placental insufficiency , 16\u201320. This study was performed to identify molecular alterations and predict dysregulated functional networks in male rat pups during (PND 1) and at the end of nephrogenesis (PND 7) to generate hypotheses which mechanisms may contribute to adverse developmental kidney programming after IUGR of different origins. We focused on the identification of common mechanisms first, followed by the analysis of model-specific alterations. Since not only LIG and LP but also intrauterine stress (IUS) induced by sham operation results in moderate IUGR and programming, we included IUS pups as a third experimental group . All expAll procedures were conducted in accordance with the German regulations and legal requirements. The experimental protocol was approved by the Institutional and Governmental Review Boards (LANUV NRW AZ 84-02.04.2012.A316).Direct comparison of kidneys during early postnatal life from IUGR offspring after 1) low protein (LP) diet throughout pregnancy , 19, (2) low protFor transcriptome microarray analysis, we randomly selected five male pups representing five different original litters per group with a bodyweight within the range of \u00b1\u20091 standard deviation around the mean weight in the respective group.For RNA and protein isolation, the whole kidney was processed using the NucleoSpin\u00ae RNA/Protein Kit according to the manufacturer. RNA concentrations were determined spectrophotometrically, and sample quality was ensured by RNA integrity number measurements.p\u2009<\u20090.05), we performed Mann-Whitney tests as post-hoc tests for all possible group comparisons . Protein and histological data were analyzed similarly. Statistical analyses were performed using GraphPad Prism 6 . All data are shown as mean \u00b1 SEM.Weight data were tested for normal distribution and, thereafter, by Grubb\u2019s test for single outliers. Afterwards, Kruskal-Wallis tests were performed as global tests for comparison of data of all four groups. In case of significance .Whole-genome expression array (Affymetrix rat gene 2.0 ST) was performed in cooperation with the Cologne Center for Genomics . Data were processed by bioinformatics to define signal strength for each transcript in each sample and the array quality. For original gene expression lists, please see NCBI Gene Expression Omnibus (GEO) archive GSE107847. Micro-RNAs were digitally excluded from further analyses, since the RNA/Protein Kit does not extract micro-RNAs. Then, datasets of fold change and p value <\u20090.05 and a fold change \u2265\u2009|1.5| in the comparisons LP\u2013C, LIG\u2013C, and/or IUS\u2013C both on PND 1 and 7. We did not perform Bonferroni adjustment of transcript data p values, because detection of perinatal programming proceedings needs a more subtle and sophisticated approach for identification of relevant alterations compared with cancer or damage models. Volcano plots were created using RStudio (3.5.0) for an overview of transcriptional alterations. Heatmaps were generated, results of the comparisons LP\u2013LIG, LP\u2013IUS, and LIG\u2013IUS included in the heatmaps, and all relevant alterations labeled by asterisks. To identify common transcriptional alterations in the IUGR groups, we used the heatmaps to find transcripts which were relevantly altered in all three or at least two of the comparisons LP\u2013C, LIG\u2013C, IUS\u2013C. Venn diagrams visualizing the number of overlaps between the IUGR groups were also generated. To identify model-specific alterations, we confirmed that the respective transcript was relevantly altered in one group of IUGR pups compared with the control group and in direct comparison with both other groups of IUGR pups as illustrated by the heatmaps.Step 1: Principal component analyses were calculated for the whole dataset as well as for the datasets on PNDs 1 and 7 separately to evaluate whether overall transcripts differ between developmental stages and/or the groups at the same developmental stage. Then, we identified relevantly altered single protein-coding transcripts in the IUGR groups by generating lists of transcripts with a http://www.ingenuity.com) on all relevantly altered transcripts. The filter excludes transcripts that do not have known relevance for the kidney in the IPA database. Additionally, we studied the NCBI gene records (http://www.ncbi.nlm.nih.gov/gene) to confirm relevance of each transcript for the kidney.Step 2: We applied the \u201ckidney filter\u201d provided by Ingenuity Pathway Analysis (IPA) software based on all transcripts with a p value <\u20090.05 . We used this stringent z-score cut off because we wanted to identify relevantly altered predicted upstream regulators only.Step 3: We used the IPA software to identify predicted upstream regulators [Step 4: In case of more than five relevantly altered transcripts or predicted upstream regulators, STRING interaction database analysis was performed to analyze functional enrichments . We appl2 kidney area. Western blots and histology were performed using standard procedures . In order to add relevance to our data, we performed single confirmatory western blots of proteins indicated to be relevant by upstream regulator analyses . Furthermore, the first data on cellular inflammation was generated by quantification of CD68 positive cells per mmBirth weight was significantly reduced in all three groups of IUGR pups Fig.\u00a0. On PND Principal component analyses indicated differences in global transcript expression patterns between PNDs 1 and 7 in all groups were relevantly altered on PND 1 . On PND 7, we identified 134 relevantly altered transcripts and IUS pups (n\u00a0=\u20091 transcript).Looking at single transcripts, 53 out of >\u200927,000 transcripts , nine in LIG pups (including increased Ren and Ccl20), and two in IUS pups (including increased Lcn2) , ten in LIG (including reduced Nupr1), and 15 in IUS pups in all groups of IUGR pups and of the nuclear transcriptional regulator protein 1 (Nupr1) in LP and LIG pups suggest increased susceptibility to monocyte infiltration and tissue remodeling [Ccl20) and S100a8/a9 as well as inhibition of the kidney androgen-regulated protein (Kap) transcript were identified in LIG pups at PND 1. CCL20 is a chemotactic factor that may aggravate renal inflammation [S100a8/a9 and decreased Kap further suggest increased susceptibility to inflammation [Ifna4) and leukotriene B4 receptor 2 (Ltb4r2) on PND 7 further hinted at dysregulation of inflammatory processes. LTB4R2 is a cell surface receptor that can bind several arachidonic acid metabolites and has stimulatory effects on neutrophils [Indeed, there is evidence for dysregulation of inflammatory processes in all three groups of IUGR pups. On PND7, reduced transcription of the paired immunoglobin-like type 2 receptor alpha in LIG and reduction of hedgehog interacting protein (Hhip) in IUS pups, both of which are known to be essential during kidney development [Beyond inflammation, we identified further common and/or group-specific molecular dysregulations during kidney development after IUGR. Dysregulation of AMPK and PPAR signaling pathways as suggested by enrichment analysis of predicted upstream regulators in LP pups on PND 7 is of interest, since this may increase the susceptibility towards obesity-associated kidney damage . PPARs helopment , 48.Our study has some methodological limitations. First, we did not perform full Bonferroni adjustment of array data. Detection of perinatal programming proceedings, however, probably needs a more subtle and sophisticated approach for gene and network identification since the expected range of change in gene expression is less pronounced than in cancer or damage models. Second, array probe identifications and the IPA database are continuously refined. Last, we had to use total kidneys for our measurements, potentially masking tissue compartment-specific alterations.In summary, dysregulation of inflammation-related pathways could be a \u201ccommon\u201d molecular feature of renal programming after IUGR of different origins although there are group- and time point-specific differences on the single transcript level. RICTOR could be a common modulator of renal programming in LP and LIG pups. Metabolic dysregulation affecting PPAR and AMPK signaling could be an insult-specific modulator of renal programming in LP pups. Based on our data, we plan to develop specific interventions to ameliorate programmed kidney disease.Supplemental File 1(DOCX 17 kb)Supplemental File 2(DOCX 13 kb)Supplemental Table 1(DOCX 14 kb)Supplemental Table 2(DOCX 18 kb)Supplemental Table 3(DOCX 26 kb)Supplemental Table 4(DOCX 14 kb)Supplemental Table 5(DOCX 16 kb)Supplemental Table 6(DOCX 16 kb)Supplemental Table 7(DOCX 17 kb)Supplemental Table 8(DOCX 15 kb)Supplemental Table 9(DOCX 22 kb)Supplemental Table 10(DOCX 21 kb)Supplemental Table 11(DOCX 21 kb)Supplemental Figure 1(A) Principal component analysis of pups on postnatal day (PND) 1 versus PND 7. Red symbols represent PND 1 pups (bordered in light red at lower left), blue symbols represent PND 7 pups (bordered in light blue at upper right). Squares represent group C, circles group LP, triangles group LIG, diamonds group IUS. There is one outlier in group LIG on PND 7 (at bottom right). Group IUS separates from the other groups on PND 7 (in the lower right within the blue border). (B) Additional principal component analysis of PND 7 pups only. The analysis confirms that group IUS (bordered in light blue) separates from all other groups (PNG 238 kb)High resolution image (TIF 27204 kb)Supplemental Figure 2p\u2009<\u20090.05; fold change \u2265 |1.5|) transcripts in the groups LP, LIG and IUS, each compared to the control group on (A) postnatal day (PND) 1 and (B) PND 7 (PNG 112 kb)Venn diagrams showing overlaps of significantly and relevantly altered (High resolution image (TIF 26557 kb)Supplemental Figure 32) are shown at the end of each line (n\u2009=\u20095\u20137 per group). Scale bar (200\u00a0\u03bcm) is shown in the lower right of each image (PNG 479 kb)Representative images of kidney sections stained with CD68 (red) and DAPI (blue) are shown for each group on postnatal day (PND) 1 (upper row of image) and PND 7 (lower row of image). Appropriate quantitative data of CD68 positive signal per kidney area (counts/mmHigh resolution image (TIF 29357 kb)"} +{"text": "Systolic left ventricular ejection fraction (LVEF) is a strong prognosticator after acute myocardial infarction (AMI). Currently, section summation of contiguous cardiac magnetic resonance (CMR) short axis steady-state free precession (SSFP) images is the reference standard for measuring LVEF.However, the use of standard CMR LVEF measurement in the early postinfarction phase is limited by the time need of image acquisition in multiple breath-holds, which is frequently not well tolerated by critically ill patients. Additional time is needed for calculating LVEF by summation of discs by semi-automated post-processing tools.Cine imaging accelerated by Temporal Parallel Acquisition Technique (TPAT) allows for the acquisition of multiple cine images in a single breath-hold. LVEF calculation from TPAT images in short an long axis orientation can be performed by a novel software tool applying four dimensional guide point modeling ventricular function analysis (4DVF). As 4DFV is based on geometric assumptions of a normal heart, it is unclear whether 4DFV is applicable to patients with AMI.To compare 4D guide-point modeling LVEF analysis of TPAT images with standard LVEF analysis in patients with AMI.27 consecutive patients underwent CMR on a 1.5 Tesla MRI scanner within 1 to 15 (mean 4) days after AMI.The CMR protocol included SSFP cine imaging in 7\u201316 (mean 13) breath-holds and inversion recovery (IR) delayed enhancement imaging following gadolinium contrast administration.Contiguous short axis cine images were analyzed by standard left ventricular function (LVF) analysis and the disc summation method .Additionally, two long axis and four short axis cine images were acquired in a single breath-hold using a TPAT accelerated SSFP sequence . The resulting cine images were used for 4-dimensional guide point modeling LVF analysis .Both LVEF calculation methods were performed by two independent observers, and the duration of analysis was recorded.Mean infarct size was 17 \u00b1 12% of left ventricular mass. 6 \u00b1 3 (range 1 to 12) LV myocardial segments were affected by wall motion abnormalities. 11 patients had anterior, 16 inferior wall myocardial infarction. A strong correlation of EF values from 4DVF with standard LVF analysis was observed . The results for observer A are shown in Table 4DVF guide point modeling is a time saving alternative to standard LVF analysis. On the basis of images acquired in a single breath-hold, it yields results that are in good accordance with standard LVF analysis.A protocol combining TPAT accelerated single breath-hold imaging and 4DVF image processing may be potentially useful for the rapid assessment of LV function in critically ill patients following acute myocardial infarction."} +{"text": "The human umbilical cord mesenchymal stem cell (hUMSC) transplantation has been shown an effective method to improve the ovarian function in POI rat model; however, the exact mechanisms are still unclear. The purpose of this study is to investigate whether the recovery of ovarian function in POI rats is related to the inhibition of tissue fibrosis following hUMSC transplantation. Furthermore, the transforming growth factor-\u03b21 and Smad3 signals was measured by Western blot and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis.The primary ovarian insufficiency (POI) rat model was established by intraperitoneal injection of chemotherapy drug cisplatin (CDDP) for 7\u2009days. The levels of serum sex hormones were measured using enzyme-linked immunosorbent assay (ELISA). The tissue fibrosis in the ovary was examined using Masson staining and Sirius red staining. The collagen fibers in the ovarian tissues were detected by Western blot analysis. To investigate the mechanisms of ovarian function recovery following hUMSC transplantation, ovarian stromal cells were isolated from the ovarian cortex of immature rats. The expression of Cytochrome P450 17A1 (Cyp17a1) and fibrosis marker of alpha smooth muscle actin (\u03b1-SMA) in ovarian stromal cells was examined using immunofluorescence analysis. Also, the protein levels of Cyp17a1 and \u03b1-SMA in ovarian stromal cells were examined by Western blot analysis. The expression of TGF-\u03b21 and p-Smad3 protein expression was observed in hUMSC-treated POI rats. The treatment with TGF-\u03b21 inhibitor of SB431542 further confirmed this signal pathway was involved in the process.The results show that the function of the ovary in POI rats was significantly improved after hUMSC transplantation. The expression of fibrosis markers (\u03b1-SMA) and production of Collagen Type I (Collagen I) and Collagen Type III (Collagen III) in POI rats were significantly inhibited in POI rats following hUMSC transplantation. In the cultured ovarian stromal cells, the decrease of TGF-\u03b21/Smad3 signaling pathway was involved in the inhibition of ovarian tissue fibrosis, which contributed to the restoration of ovarian function in POI rats following hUMSC transplantation.Our study demonstrated that the TGF-\u03b2"} +{"text": "Ileal neuroendocrine tumors (NETs) represent the most common neoplasm of the small intestine. Although up to 50% of patients with ileal NETs are diagnosed with multifocal disease, the mechanisms by which multifocal ileal NETs arise are not yet understood. In this study, we analyzed genome\u2010wide sequencing data to examine patterns of copy number variation in 40 synchronous primary ileal NETs derived from three patients. Chromosome (chr) 18 loss of heterozygosity (LOH) was the most frequent copy number alteration identified; however, not all primary tumors from the same patient had evidence of this LOH. Our data revealed three distinct patterns of chr18 allelic loss, indicating that primary tumors from the same patient can present different LOH patterns including retention of either parental allele. In conclusion, our results are consistent with the model that multifocal ileal NETs originate independently. In addition, they suggest that there is no specific germline allele on chr18 that is the target of somatic LOH. However, up to 50% of patients with ileal NETs are diagnosed with multiple synchronous primary tumors.22.1Our sample set consisted of 40 de\u2010identified synchronous primary tumors and matched adjacent normal ileal mucosa specimens from three ileal NET patients Figure . A piece2.2Genomic DNA (gDNA) was extracted from fresh frozen tumor and adjacent normal ileum samples. Tissue specimens from patient 1 were sequenced using the 10\u00d7 Genomics linked\u2010read WGS approach , as detailed before.2.3https://github.com/gavinha/TitanCNA_10X_snakemake),For patient 1, somatic copy number changes and LOH in tumor samples were detected using a modified TITAN pipeline retaining tumor samples that have at least 1000 LOH\u2010informative SNPs or at least 100 LOH\u2010informative SNPs for further analysis; (f) assigning the retained allele of each LOH\u2010informative SNP by comparing read counts of the reference and alternative alleles (Table Detailed LOH mapping was performed for tumors showing chr18 LOH by completing the following steps: (a) identifying germline heterozygous SNPs in the normal samples using the following filters: read depth >10 and variant allele frequency between 0.4 and 0.6es Table .33.1Copy number analysis identified hemizygous loss of chr18 (log2 [copy number/2]\u2009<\u2009\u20110.1) to be the most common genomic alteration in multifocal ileal NETs Figure , consist3.2By comparing retained SNP alleles between tumor samples with chr18 LOH, we observed three different chr18 LOH patterns in multifocal ileal NETs. Individual tumors with chr18 allelic loss from the same patient can lose either (a) the same parental allele, (b) different parental alleles with consistent allelic loss across chromosomal arms, or (c) different parental alleles in the short (p) and long (q) arms of chr18 Figure . Copy nu4Ileal NETs represent the most common neoplasm of the small intestine with a high incidence of multiple synchronous primary tumors at diagnosis. Our understanding of the tumor multifocality, however, remains limited with respect to the molecular mechanisms underlying their tumorigenesis and effects on clinical management.Most large\u2010scale sequencing studies to date have concentrated on sequencing single primary tumors from ileal NET patients.The origin of multifocal ileal NETs is not well understood. Previous studies have proposed that multifocal ileal NETs may arise due to germline predisposition or as independent clones.The significance of our study is three\u2010fold. First, consistent with previous results, we show that chr18 loss occurs in many, but not all, primary ileal NETs.There are some limitations in our current study. First, our interpretation of LOH pattern assumed that the hemizygous loss of chr18 in ileal NET is more likely to occur in a single parental allele as previously reported.In conclusion, our data demonstrate that chr18 LOH is commonly found in multifocal ileal NETs. The distinct LOH patterns in individual tumors from the same patients suggest that there is not a particular germline allele on chr18 that predisposes somatic LOH.The authors declare that they have no conflicts of interest with the contents of this article. However, Dr Meyerson declares the following general conflicts of interest: research support from Bayer, Ono, Novo, and Janssen; patent licensing royalties from LabCorp; and serving as scientific advisory board chair and consultant for OrigiMed.Figure S1 Copy number profiles of all primary synchronous ileal NETsClick here for additional data file.Table S1 Patient informationClick here for additional data file.Table S2 Summary of SNP filtering for LOH mappingClick here for additional data file.Table S3 Summary table of chr18 LOH patterns for three patientsClick here for additional data file.Table S4 Retained alleles of annotated SNPs of tumors displaying chr18 LOH in patient 1Click here for additional data file.Table S5 Retained alleles of annotated SNPs of tumors displaying chr18 LOH in patient 2Click here for additional data file.Table S6 Retained alleles of annotated SNPs of tumors displaying chr18 LOH in patient 3Click here for additional data file."} +{"text": "PRNP) and account for about 15% of human prion disease cases worldwide. The proposed mechanism is that the mutation predisposes to conformational change in the expressed protein, leading to the generation of disease-related multichain PrP assemblies that propagate by seeded protein misfolding. Despite considerable experimental support for this hypothesis, to-date spontaneous formation of disease-relevant, transmissible PrP assemblies in transgenic models expressing only mutant human PrP has not been demonstrated. Here, we report findings from transgenic mice that express human PrP 117V on a mouse PrP null background (117VV Tg30 mice), which model the PRNP A117V mutation causing inherited prion disease (IPD) including Gerstmann-Str\u00e4ussler-Scheinker (GSS) disease phenotypes in humans. By studying brain samples from uninoculated groups of mice, we discovered that some mice (\u2265475 days old) spontaneously generated abnormal PrP assemblies, which after inoculation into further groups of 117VV Tg30 mice, produced a molecular and neuropathological phenotype congruent with that seen after transmission of brain isolates from IPD A117V patients to the same mice. To the best of our knowledge, the 117VV Tg30 mouse line is the first transgenic model expressing only mutant human PrP to show spontaneous generation of transmissible PrP assemblies that directly mirror those generated in an inherited prion disease in humans.Inherited prion diseases are caused by autosomal dominant coding mutations in the human prion protein (PrP) gene ( Transgenic mice expressing the human prion protein containing a mutation linked to the inherited prion disease Gerstmann-Str\u00e4ussler-Scheinker disease develop spontaneous neuropathology. This represents the first human prion protein transgenic model to show spontaneous generation of transmissible prion assemblies that directly mirror those generated in humans. Sc . ThP 27\u201330) . CollectSpont-A mouse brain).Spontaneous 117V prion isolates showed no major alteration of their transmission properties after secondary passage in 117VV Tg30 mice . A totalSc assembly [Spont-A mouse, its brain showed only focal 117V PrP plaques at a level very much lower than present in the brain of subclinically infected mice from the primary or secondary transmission series. These observations suggest that 117V PrP plaque load is dissociated from a clinical phenotype. Preferential propagation of the stable 117V PrP amyloid assembly rather than the labile 117V PrPSc assembly may therefore result in a progressive clinically silent PrP amyloidosis. Testing the validity of this hypothesis and whether efficient propagation of labile 117V PrPSc is responsible for clinical phenotype will now require the development of new methods for isolation of each distinct 117V PrP assembly.One possible explanation for this apparent lack of adaptation may relate to the relative efficiencies of replication of the 2 disease-related 117V PrP assemblies that are propagating in these transmissions. Although accurate measurement of the concentration of the stable 117V PrP amyloid assembly would be possible, the inherent instability of the labile 117V PrPassembly precludeSc or distinct PrP amyloid fibres) generated from either mutant or wild-type PrP, combined with differences in their regional distribution, abundance, and potential neurotoxicity, provides a complex molecular mechanism for generating phenotypic heterogeneity in patients with the same PRNP mutation.Variable conversion of wild-type PrP to pathogenic conformations is thought to be an important contributor to phenotypic variability in certain IPDs associated with different mutations ,29,66\u201368Sc. These data suggest that the stable 117V PrP assembly that generates the 8 kDa proteolytic fragment and forms amyloid plaques in 117VV Tg30 mice can also template the misfolding of wild-type human PrP 129V to a similar amyloid conformation. Based upon these findings, primary transmissions of A117V patient brain to mice expressing wild-type human PrP should be revisited and compared to the transmission properties of the same A117V patient isolates after passage in 117VV Tg30 mice to see if the transmission frequency of clinically silent amyloid phenotypes is similar.Accordingly, in further transmission experiments, we examined whether spontaneous 117V prion isolates passaged once in 117VV Tg30 mice could also propagate in transgenic mice expressing wild-type human PrP 129V . SurprisPRNP point mutations. We found that the mean age of clinical onset in IPD A117V patients was significantly earlier than seen in either IPD P102L or E200K patients are distinct from amyloid PrP assemblies that characterise the GSS IPD neuropathological phenotype [These findings are consistent with the highly distinct transmission properties of similar disease-associated PrP assembly states from GSS patients with the P102L mutation \u201335 and shenotype ,38.To the best of our knowledge, 117VV Tg30 mice represent the first authentic transgenic model capable of recapitulating spontaneous prion disease pathogenesis seen in IPD, and these findings significantly support the protein-only hypothesis of prion propagation. Although other models that have superimposed human mutations onto rodent PrP are also capable of spontaneous formation of infectious PrP assemblies \u201354, our The spontaneous generation of infectious 117V PrP assemblies in transgenic mice that we describe here provides important new insight into disease pathogenesis in IPD A117V patients. As our previous transgenic models of IPD P102L and E200K failed to generate spontaneous abnormal PrP assemblies, we speculated that the kinetics of spontaneous PrP misfolding associated with the A117V mutation may be more rapid, thereby enabling observation of de novo initiation of prion pathogenesis within the lifespan of a mouse. Comparison of the mean ages of clinical disease onset in patients strongly supports this idea, because IPD A117V patients have a significantly earlier age of onset than patients with IPD P102L or E200K. Moreover, the significant effect of codon 129 zygosity in modulating the age of disease onset seen in IPD A117V patients can now be reasonably explained by our finding that spontaneously generated 117V infectious PrP assemblies can convert wild-type human PrP 129V to a protease-resistant disease-related PrP assembly. However, it is important to mention that involvement of an additional mechanism for this effect cannot be excluded, because we have not yet examined whether wild-type human PrP 129M has partial dominant-negative inhibitory effects ,76,81,82In summary, we report the first model of IPD that spontaneously generates both human prions and PrP amyloid. Our demonstration that 117VV Tg30 mice faithfully recapitulate IPD A117V molecular and neuropathological phenotypes suggests that these mice may be useful for future translational studies. Comparison of treatment groups with untreated aged cohorts of 117VV Tg30 mice should have the statistical power to inform upon the early or presymptomatic pathogenesis of disease and efficacy of long-term treatments that may prevent spontaneous prion disease. Treatments have or are being developed that target normal PrP \u201385, althwww.nc3rs.org.uk/ARRIVE/).Storage and biochemical analyses of post mortem human brain samples and transmission studies to mice were performed with written informed consent from patients with capacity to give consent. When patients were unable to give informed consent, assent was obtained from their relatives in accordance with UK legislation and Codes of Practice. Samples were stored and used in accordance with the Human Tissue Authority Codes of Practice and in line with the requirements of the Human Tissue Authority licence held by UCL Institute of Neurology. This study was performed with approval from the National Hospital for Neurology and Neurosurgery and the UCL Institute of Neurology Joint Research Ethics Committee \u2014REC references: 03/N036, 03/N038 and 03/N133. Work with mice was performed under approval and licence granted by the UK Home Office Act 1986); Project Licence number 70/6454, which conformed to University College London institutional and ARRIVE guidelines designated Tg(HuPrP117V 129V+/+Prnpo/o)-30 mice (117VV Tg30) have been described previously [129V transgene array and murine PrP null alleles (Prnpo/o) designated Tg(HuPrP129V+/+Prnpo/o)-152 mice (129VV Tg152) have been described previously [Transgenic mice homozygous for a human PrPeviously ,20,69\u2013722 and intracerebrally inoculated into the right parietal lobe with 30 \u03bcl of 1% (w/v) brain homogenate prepared in Dulbecco\u2019s phosphate buffered saline lacking Ca2+ and Mg2+ ions (D-PBS). All mice were then examined daily for early indicators of clinical prion disease, including piloerection, sustained erect ears, intermittent generalised tremor, unsustained hunched posture, rigid tail, mild loss of coordination, and clasping hind legs when lifted by the tail. Definite diagnosis of clinical prion disease was reached if mice exhibited any two early indicator signs in addition to one confirmatory sign or any two confirmatory signs. The confirmatory signs included ataxia, impairment of righting reflex, dragging of hind limbs, sustained hunched posture, or significant abnormal breathing. Mice were killed (by CO2 asphyxiation) if they exhibited any signs of distress or once a diagnosis of prion disease was established. Brains from inoculated mice were removed at autopsy, divided sagittally with half frozen, and half fixed in 10% buffered formol saline. Subsequent immunohistochemical or biochemical investigations were performed blind to sample provenance.Strict bio-safety protocols were followed. Inocula were prepared, using disposable equipment for each inoculum, in a microbiological containment level 3 laboratory and inoculations performed within a class 1 microbiological safety cabinet. For primary transmission and serial passages, groups of animals comprising 117VV Tg30 and 129VV Tg152 mice were inoculated with appropriate mouse brain homogenates. Details of primary transmissions of A117V patient brain samples to 117V Tg30 mice are described in the work by Asante and colleagues [Brains were fixed in 10% buffered formalin for at least 48 hours and then immersed in 98% formic acid for 1 hour, postfixed in formalin, and then processed for paraffin wax embedding. Serial sections of 4-\u03bcm thickness were pretreated by boiling for 10 minutes in a low ionic strength buffer before exposure to 98% formic acid for 5 minutes. Abnormal PrP accumulation was examined using anti-PrP monoclonal antibodies 3F4 or ICSM Sc from 10% (w/v) brain homogenates (117VSpont-A and 117VSpont-B isolates only) was performed using established procedures [Preparation of brain homogenates (10% (w/v) in D-PBS), PK digestion , and subsequent immunoblotting using high-sensitivity chemiluminescence was performed as described previously ,63. NaPTocedures ,63. Blotocedures ,63. PrimS1 FigFixed brain samples from the cohort of aged uninoculated 117VV Tg 30 mice shown in (TIF)Click here for additional data file.S2 FigSpont-A was culled with neurological disease at 476 days of age. (B) The original uninoculated 117VV Tg30 designated 117VSpont-B was culled without neurological symptoms at 734 days of age. Both 117VSpont-A and 117VSpont-B mice had spontaneous PrP plaques in the anterior commissure of brain. In the transmission series reported, 1% (w/v) brain homogenate was used for all inoculations. Mice were observed for clinical signs of prion disease using criteria described in Materials and methods. Post mortem brain from inoculated mice was examined for evidence of abnormal PrP propagation by IB and/or IHC examination. PrP, prion protein; IB, immunoblotting; IHC, immunohistochemistry(A) The original uninoculated 117VV Tg30 mouse designated 117V(TIF)Click here for additional data file.S3 FigThe panels show schematic drawings reflecting the overall spatial distribution and intensity of the gliosis or PrP deposition within the experimental groups. They are not meant to indicate precise representations of individual brains. * Definition of values for neuronal loss: NL0 (none): No neuronal loss; NL+ (mild): Drop out of single neurones either focally or within the Ammon\u2019s horn (AH), leaving the AH continuity intact; NL++ (moderate): Focal or regional drop out, interrupting the continuity of the AH and creating a small to medium gap (up to 1/3 of the length of the AH); NL+++ (severe): Neuronal drop out leaving gaps of more than 1/3 of the AH\u2019s length. Ratios represent the proportion of samples with the corresponding neuronal loss score. Note that gliosis variability within experimental groups is considerable. PrP, prion protein(TIF)Click here for additional data file.S4 FigDetails of these transmissions are shown in (TIF)Click here for additional data file.S1 DataPrP, prion protein(XLSX)Click here for additional data file."} +{"text": "Objective: Screening for novel prognostic biomarkers and potential therapeutic targets from colorectal cancer microenvironment.Results: 372 genes were overexpressed in colorectal cancer microenvironment, five of which that had the most prognostic powers were enriched in Epithelial-Mesenchymal Transition and cell cycle pathways. For the first time, we showed that SMARCD3 was mainly expressed in CAFs and could be a novel prognostic marker and potential therapeutic target. Function analyses indicated that MSARCD3 might promote CAFs activation and colorectal cancer metastasis through SMARCD3-WNT5A/TGF-\u03b2-MAPK14-SMARCD3 positive feedback loop. Signaling map of SMARCD3 was constructed and several potential drugs that could regulate SMARCD3 were also presented.Conclusions: SMARCD3 is a novel prognostic biomarker and potential therapeutic target of colorectal cancer, which may promote cancer metastasis through activation of CAFs.Methods: Colorectal cancer microenvironment related genes were screened based on immune and stromal scores. Function enrichment analyses were performed to show the underlying mechanistic insights of these tumor microenvironment related genes. Kaplan-Meier survival analysis was used for evaluating the prognostic power. Gene-Pathway interaction network analysis and cellular heterogeneity analysis of tumor microenvironment were also performed. Gene set enrichment analysis was performed for signal gene pathway analysis. Protein data from The Cancer Genome Atlas were used for validation. Colorectal cancer is one of the most common digestive malignancies worldwide . The tumNevertheless, the molecular mechanisms underlying TME associated colorectal cancer progression have never been well elucidated. Hence, screening for novel prognostic biomarkers and potential therapeutic targets for colorectal cancer from TME is of crucial importance. This study takes advantage of publicly available datasets and powerful bioinformatics tools to screen for genes with significant prognostic value and explore potential mechanistic insights.Immune score and stromal score of each sample were computed using ESTIMATE algorithm. TME related DEGs were computed based on immune and stromal scores using Agilent microarray expression data and RNAseq data of colon cancer from TCGA. The heat maps of DEGs based on different grouping strategies were presented in Gene ontology and KEGG analyses of these 372 genes were performed using network analyst \u20131E. ProtPrognostic power of 372 commonly upregulated genes in TME were evaluated through Kaplan-Meier survival analysis using colorectal cancer data from TCGA. Five genes with most significant prognostic power were selected for further analyses. The expression levels of SMARCD3, CRIP2, PRAM1, HSPB2 and CERCAM in Agilent stromal/immune high/low groups and RNAseq stromal/immune high/low groups were demonstrated in To explore the association among these fives genes and different cell types in the tumor microenvironment, cellular heterogeneity analyses of tumor microenvironment were performed using xCell using ssGSEA method. The correlation map of the expression value of five genes and enrichment score of different cell components in TME were shown in high) > PAS (gene Xlow) indicates gene X has an activation effect, otherwise an inhibition effect. Analyses results indicated that SMARCD3, CRIP2, PRAM1, HSPB2 and CERCAM were associated with epithelial-mesenchymal transition (EMT) pathway activation of 10 cancer related pathways in 32 cancer types were computed based on RPPA protein data from TCGA. PAS , which iAs we mentioned in the above section, SMARCD3 was associated with cancer metastasis and EMT related gene signatures. Further PPI analysis indicated that SMARCD3 might promote colorectal cancer metastasis through MAPK14, MYOD1 or SMAD4 related pathways. Previously, it was reported that SMARCD3 could stimulate EMT of breast cancer cells through upregulating WNT5A expression . While WUsing TCGA colorectal cancer RNAseq and protein expression data, we demonstrated that WNT5A and TGFBIn this study, we found 372 genes that overexpressed in TME based on immune score and stromal score using TCGA COAD data from two platforms (Agilent and RNAseq). GO and KEGG pathway enrichment analyses showed that these 372 genes were enriched in immune response, cytokine production and toll-like receptor signaling pathway etc.. SMARCD3, CRIP2, PRAM1, HSPB2 and CERCAM were selected for further analyses due to their most significant prognostic powers. Cellular heterogeneity analysis indicated that PRAM1 was associated with macrophages while SMARCD3, CRIP2, HSPB2 and CERCAM were correlated with fibroblasts. Pathway analyses showed that the five genes were involved in EMT activation and cell cycle inhibition. Since EMT was an important factor for CAFs activation, the above results implicated a potential role of these genes in inducing CAFs activation.CAFs plays critical roles in tumor proliferation, invasion, angiogenesis and regulation of tumor immune microenvironment. For instance, CAFs could release exosomes, VEGF , HGF (hepatocyte growth factor) and GAS6 (growth arrest-specific gene 6) to promote cancer proliferation and invasion, and affect the function of epithelial cell and macrophages ; regulatSMARCD3 encoded protein belongs to SWI/SNF family which display helicase and ATPase activities and could regulate gene transcription by altering the chromatin. It was reported that knock down SMARCD3 expression could induce mesenchymal-epithelial transition (EMT) of breast cancer cells . Here, wBased on the above findings, we speculated that SMARCD3-WNT5A/TGF-\u03b2-MAPK14-SMARCD3 positive feedback loop might be activated in fibroblasts and play critical roles in promoting CAFs activation and cancer metastasis . Expression profiles of colon, polyp and primary colon cancer were obtained from Gene Expression Omnibus ). Expression of SMARCD3 in different clinical groups and its correlation with methylation were plotted using UALCAN based on data from TCGA [Gene-pathway interaction network analysis was performed using GSCAlite . Brieflyrom TCGA . The hearom TCGA .https://rpkgs.datanovia.com/survminer/index.html) and Kaplan-Meier analyses were performed through survival package [Risk score of each sample was computed based on expression value of the 5 genes using cox proportional hazard model. The best cutoff value of 5 gene risk score was computed using survminer package ( package in R. Al package . SignifiData sharing is not applicable to this article as no new data were created or analyzed in this study.Supplementary Table 2Supplementary Table 1Supplementary Table 6Supplementary Tables 7 and 8Supplementary Tables 3, 4 and 5Supplementary Figures"} +{"text": "Cancers launched the topic collection \u201cDrug Resistance and Novel Therapies in Cancers\u201d in the second half of 2018.Cancer is among the leading causes of mortality in the world, accounting for an estimated 9.6 million deaths in 2018. Despite significant advances in cancer therapy, drug resistance remains the principal limiting factor in achieving cures in patients with cancers. Scientific communities have been working intensively to understand the molecular mechanisms underlying drug resistance and to develop novel therapies to overcome drug resistance. Under these circumstances, In the last four months of 2018, eight cutting-edge research articles and comprehensive review papers had been published in this topic collection. One research article explored the possibility of increasing the sensitivity of triple negative breast cancer (TNBC) cells to hormonal therapy [Two research articles were related to melanoma ,3. One aAnther research aimed to indirectly target rapamycin complex 1 (mTORC1) hyperactivity in cancers by exploiting homeostatic vulnerabilities within Tuberous Sclerosis Complex 2 (TSC2)-deficient cells . Both chIonizing radiation (IR) has been an important treatment for colorectal cancer (CRC). However, IR activates nuclear factor kappa B (NF-\u03baB) that causes radio-resistance and stimulates matrix metalloproteinase (MMP)-2/-9, which promote tumor migration and invasion. The research by Sugano et al. examinedThree published reviews covered three important areas of cancer therapy. One review article covered the latest advances in a hepatocellular carcinoma therapy that is a rapidly evolving . While t"} +{"text": "Monoclonal antibodies can mediate protection against Ebola virus (EBOV) infection through direct neutralization as well as through the recruitment of innate immune effector functions. However, the antibody functional response following survival of acute EBOV disease has not been well characterized. In this study, serum antibodies from Ebola virus disease (EVD) survivors from Sierra Leone were profiled to capture variation in overall subclass/isotype abundance, neutralizing activity, and innate immune effector functions. Antibodies from EVD survivors exhibited robust innate immune effector functions, mediated primarily by IgG1 and IgA1. In conclusion, development of functional antibodies follows survival of acute EVD. The humoral immune response in human survivors of Ebola virus disease includes the development of neutralizing antibodies and polyfunctional IgG1 and IgA antibodies that can mediate effector functions against the Ebola virus glycoprotein via multiple innate immune effector cell types. The 2013\u20132016 Ebola virus disease (EVD) epidemic in West Africa and current outbreaks in the Democratic Republic of Congo indicate that highly pathogenic filoviruses remain a significant threat to human health. The ability of antibodies to provide protection from a lethal Ebola virus (EBOV) challenge has been demonstrated in the context of pre- and postexposure administration of EBOV glycoprotein (GP)\u2013specific monoclonal antibodies (mAbs) .EVD survivors develop both humoral and cellular immunity against several EBOV proteins, including GP, secreted GP (sGP), nucleoprotein (NP), and matrix protein VP40 . Early sPlasma samples were collected from individuals with documented clinical history of EVD approximately 6 months following survival , househoRecombinant GP, sGP, VP40 (IBT Bioservices), and NP (Sino Biologics) were coupled to MagPlex beads (Luminex) according to previously published protocols . Samples4 cells/well) for 1 hour (antibody-dependent phagocytosis by neutrophils [ADNP]) or adding THP-1 cells at 2.5 \u00d7 104 cells/well for 18 hours (antibody-dependent cellular phagocytosis by monocytes [ADCP]). Uptake of antibody-bead complexes by cells was determined by flow cytometry, and a phagocytic score was determined: (% fluorescein isothiocyanate [FITC]+ cells) \u00d7 (gMFI FITC+) / 10 000.Biotinylated GP was conjugated to yellow-green Neutravidin beads (Thermo Fisher) and incubated with samples for 2 hours prior to adding white blood cells from human donor peripheral blood (5 \u00d7 10Biotinylated GP-coated red Neutravidin beads (Thermo Fisher) were incubated with heat-inactivated samples. Guinea pig complement (Cedarlane Labs) diluted in veronal buffer containing calcium and magnesium (Boston Bioproducts) was incubated with antibody-bead complexes for 20 minutes, and C3 deposition onto beads was detected using an anti\u2013guinea pig C3 antibody and measured by flow cytometry.4 cells/well) for 5 hours with brefeldin A, GolgiStop, and anti-CD107a (BD Biosciences). Intracellular cytokine staining to detect interferon-\u03b3 and macrophage inflammatory protein-1\u03b2 (BD Biosciences) was performed using Fix/Perm (Life Technologies), and cells were analyzed by flow cytometry.Enzyme-linked immunosorbent assay (ELISA) plates were coated with GP antigen (300 ng/well). Wells were washed, blocked, and incubated with samples for 2 hours prior to adding NK cells enriched from the peripheral blood of human donors (5 \u00d7 104 renilla luciferase units/well of EBOV GP-pseudotyped vesicular stomatitis virus expressing luciferase (IBT Bioservices), followed by overnight incubation with VeroE6 monolayers. Cells were lysed and luciferase activity was determined using a luciferase-activating reagent (Promega). Endpoint neutralization titers were defined as the last dilution that reduced luciferase activity by 50% or 80% compared to control wells.Two-fold dilutions of samples (1:12\u20131:384) were incubated with 4 \u00d7 10Samples were diluted and incubated with CaptureSelect IgG1, IgA, or control resins (Thermo Fisher Scientific) for 1 hour. Depleted samples were collected by centrifugation using a 30\u201340 \u00b5m filter plate. Depletion was confirmed using a multiplexed human isotyping kit (MilliporeSigma).P < .05) were used to generate networks in Cytoscape (version 3.4.0).Associations between antibody features were determined using nonparametric Spearman correlation coefficient. Statistically significant associations after Bonferroni correction for multiple comparisons (adjusted Univariate analyses were performed using Prism 7 software. Kruskal\u2013Wallis with Dunn multiple correction test or Mann\u2013Whitney with Bonferroni correction was used to determine statistical significance between groups.We determined relative antibody levels of IgG subclasses and isotypes specific for EBOV GP, sGP, NP, and VP40 of EVD survivors, seronegative individuals, and household contacts. Survivors demonstrated higher antibody responses against GP, sGP, and NP compared to seronegative and household contact controls from the same region across several IgG subclasses and isotypes . We obseAs GP is exposed on virions and infected cells, we measured the ability of GP-specific antibodies to mediate neutralization and induce innate immune effector functions: ADNP, ADCP, ADCD, and antibody-dependent NK cell activation . The surTo determine if either neutralizing or effector function activity was associated with a specific antibody subclass and isotype, we performed a network analysis using significant associations between GP- and sGP-specific antibody levels and functional activity induced by the survivor samples . While nGiven the coordination of IgG1 and IgA responses and association with induction of innate immune effector functions, we hypothesized that IgG1 and IgA may mediate disparate functional activities. We depleted IgG1 and IgA from the samples using resins and evaluated depleted samples for induction of effector functions from neutrophils, monocytes, and NK cells. Depletion of IgG1 from the samples significantly reduced the ability of all samples to mediate functional activity compared to samples incubated with a control bead resin , indicatDepletion of IgA had a more heterogenous impact on functional activity. ADNP activity was reduced in half of the IgA-depleted samples, and 4 of the samples had reductions of ADNP activity of >40% . UnexpecAs IgA1 and IgA2 may have differential impact on antiviral immunity , we nextStudies in animal models indicates that protective antibodies use both neutralization and recruitment of innate immune effector functions to provide protection against a lethal EBOV infection. Here we show that the humoral immune response that develops in human survivors of EVD resembles that of protective monoclonal antibodies, marked by the development of both neutralizing and polyfunctional antibodies. Importantly, our analysis demonstrates that both IgG1 and IgA antibodies mediate effector functions.+ T-cell responses in the same EVD survivors evaluated here demonstrated a relatively low abundance of GP-specific CD8+ T cells, yet much higher levels of NP- and VP40-specific CD8+ T cells [+ T cells may indicate that the two branches of adaptive immunity are differentially shaped by distinct EBOV proteins that may complement each other to maximize immunity.We found that all survivors developed antibody responses against GP and sGP, consistent with previous reports , 11. Rec T cells . The splGiven the mucosal nature of the initial infection, the development functional IgA against GP may provide protection at mucosal sites. Macrophage and neutrophils within the mucosa express high levels of Fc\u03b1R , and thuThe Journal of Infectious Diseases online. Consisting of data provided by the authors to benefit the reader, the posted materials are not copyedited and are the sole responsibility of the authors, so questions or comments should be addressed to the corresponding author.Supplementary materials are available at jiz364_suppl_Supplementary_Figure_S1Click here for additional data file.jiz364_suppl_Supplementary_Figure_S2Click here for additional data file.jiz364_suppl_Supplementary_LegendsClick here for additional data file.jiz364_suppl_Supplementary_Table_S1Click here for additional data file."} +{"text": "Posttranscriptional modifications of anticodon loops contribute to the decoding efficiency of tRNAs by supporting codon recognition and loop stability. Consistently, strong synthetic growth defects are observed in yeast strains simultaneously lacking distinct anticodon loop modifications. These phenotypes are accompanied by translational inefficiency of certain mRNAs and disturbed protein homeostasis resulting in accumulation of protein aggregates. Different combinations of anticodon loop modification defects were shown to affect distinct tRNAs but provoke common transcriptional changes that are reminiscent of the cellular response to nutrient starvation. Multiple mechanisms may be involved in mediating inadequate starvation response upon loss of critical tRNA modifications. Recent evidence suggests protein aggregate induction to represent one such trigger. LysUUU contains mcm5s2U34 (5-methoxycarbonylmethyl-2-thiouridine at position 34) and ct6A37 (cyclic N6-threonylcarbamoyladenosine at position 37) modifications which each fulfill one or more of these tasks precursor requires TCD1 genes in different tRNA modification mutants genes in addition to the activation of different amino acid biosynthesis genes accumulating protein aggregates independent of a tRNA modification defect (Bruch et al. zuo1 mutants, the ribosome-associated chaperone system is severely compromised, leading to accumulation of protein aggregates (Bruch et al. Intriguingly, when studying the transcriptional induction of nutrient responsive genes in combined tRNA modification mutants, their expression was dampened upon overexpression of the very same tRNAs that conferred a suppression of growth defects (Bruch et al."} +{"text": "Scientific Reports 10.1038/s41598-020-58183-4, published online 29 January 2020Correction to: The original version of this Article contained a repeated typographical error where \u201cDifferential Mobility Analyzer\u201d and \u201cDifferential Mobility Analysis\u201d were given as \u201cdynamic mobility analyzer\u201d and \u201cdynamic mobility analysis\u201d respectively.In the Abstract,\u201cDynamic mobility analysis (DMA) indicates smaller particles are fabricated by discharging zinc wires in a higher magnetic field.\u201dnow reads:\u201cDifferential Mobility Analysis (DMA) indicates smaller particles are fabricated by discharging zinc wires in a higher magnetic field.\u201dIn the Results section,\u201cTo investigate nanoparticle distribution of aerosols created by sparking discharge and influence of magnetic field on median aerodynamic diameter we employed dynamic mobility analyzer (DMA), measured at different applied voltage and with and without presence of magnetic field.\u201dnow reads:\u201cTo investigate nanoparticle distribution of aerosols created by sparking discharge and influence of magnetic field on median aerodynamic diameter we employed Differential Mobility Analyzer (DMA), measured at different applied voltage and with and without presence of magnetic field.\u201dIn the Methods section,36,37.\u201d,\u201cIn Fig. 7 schematically is represented experimental setup of sparking machine connected to Dynamic Mobility Analyzer (DMA), operating principle was explained previouslynow reads:36,37.\u201d\u201cIn Fig. 7 schematically is represented experimental setup of sparking machine connected to Differential Mobility Analyzer (DMA), operating principle was explained previouslyIn the legend of Table 1, where,3.\u201d\u201cTable 1. Comparison of results obtained from dynamic mobility analyzer (DMA) by sparking Zinc wire at different conditions. Geometric mean as average particle size, concentration in number of particles per cmnow reads:3.\u201d\u201cTable 1. Comparison of results obtained from Differential Mobility Analyzer (DMA) by sparking Zinc wire at different conditions. Geometric mean as average particle size, concentration in number of particles per cmAdditionally, the first two sentences in the Particle distribution (DMA) section are a duplication of the legend of Table 1. The text,3.\u201d, has been removed from this section.\u201cTable 1. Comparison of results obtained from dynamic mobility analyzer (DMA) by sparking Zinc wire at different conditions. Geometric mean as average particle size, concentration in number of particles per cmThese errors have now been corrected in the PDF and HTML versions of the Article."} +{"text": "TP53 mutation or copy number loss was identified in all evaluable tumors with evidence of C19MC overexpression. We find that C19MC miRNAs exhibit significant negative correlation to TP53 regulatory miRNAs and K-Ras regulatory miRNAs. Using RNA-seq we identified that pathways relevant to cellular differentiation as well as mRNA translation machinery are transcriptionally enriched in UESL. In summary, utilizing a combination of next-generation sequencing and high-density arrays we identify the combination of C19MC hyperexpression via chromosomal structural event with TP53 mutation or loss as highly recurrent genomic features of UESL.Undifferentiated embryonal sarcoma of the liver (UESL) is a rare and aggressive malignancy. Though the molecular underpinnings of this cancer have been largely unexplored, recurrent chromosomal breakpoints affecting a noncoding region on chr19q13, which includes the chromosome 19 microRNA cluster (C19MC), have been reported in several cases. We performed comprehensive molecular profiling on samples from 14 patients diagnosed with UESL. Congruent with prior reports, we identified structural variants in chr19q13 in 10 of 13 evaluable tumors. From whole transcriptome sequencing, we observed striking expressional activity of the entire C19MC region. Concordantly, in 7 of 7 samples undergoing miRNAseq, we observed hyperexpression of the miRNAs within this cluster to levels >100 fold compared to matched normal tissue or a non-C19MC amplified cancer cell line. Concurrent TP53 mutation or copy number loss. Therefore, we provide for the first time a genomic landscape of UESL and describe cellular consequences of the discovered foundational genomic changes.We perform the most comprehensive molecular analysis to date of undifferentiated embryonal sarcoma of the liver, utilizing whole exome sequencing, RNA sequencing, miRNA sequencing, and SNP arrays. We find that UESL tumors harbor aberrant transcriptional start sites within C19MC region that is driven by structural rearrangement, leading to extreme overexpression of C19MC miRNAs. We further find that C19MC miRNAs negatively correlate with TP53 and K-Ras regulatory miRNAs. UESL also harbors highly recurrent Undifferentiated embryonal sarcoma of the liver (UESL) is an aggressive primitive malignancy. It occurs predominantly in children with a peak age incidence of 6\u201310 years and equally across gender , presentMALAT1 to 19q13.4 being the most recurrently reported [Though the oncogenesis of UESL remains uncertain, in some cases UESL can arise from malignant transformation of mesenchymal hamartoma of the liver (MHL) \u20137. Supporeported . Given tThe chromosome 19 miRNA cluster has recently gained interest as a potential oncomir. C19MC has been implicated in various tumors including but not limited to embryonal tumors with multilayered rosettes , hepatocTP53 mutation or loss is present in all samples that also display C19MC changes. In light of previous literature and the highly recurrent nature of these events, our study provides compelling evidence that these two genetic events are foundational and perhaps pathognomonic of this disease.The primary objective of our study was to perform a survey of the genomic landscape of UESL to determine the most common recurrent molecular alterations. Using patient samples, we perform a combination of whole exome sequencing, RNA sequencing, miRNA sequencing, and single nucleotide polymorphism (SNP) arrays to evaluate for recurrent mutations, copy number changes, and expressional events. We find that UESL display a highly aneuploid genome with recurrent structural alterations of chr19q13 that are uniformly associated with aberrantly high levels of transcriptional activity of the chromosome 19 microRNA cluster. In addition we find that Fourteen subjects with a diagnosis of undifferentiated embryonal sarcoma of the liver were identified including 5 males and 9 female with median age of 8 years at diagnosis (range 8mo\u2013 11 years). Sequencing studies were performed where quality and quantity of nucleic acids allowed and included either tumor-normal matched (7) or tumor only (7) whole exome sequencing, SNP arrays (13), RNAseq (13) and miRNAseq (7). Additional demographic and sequencing details as well as summary findings are provided in Whole exome sequencing generated a median 136 Million (M) total read pairs, resulting in 107x coverage (range 67x-200x) across the capture region after duplication removal and mapping. RNA sequencing was performed with an average yield of 82M read pairs (range 58-97M). RNAseq data from the one tumor taken from FFPE materials failed quality control and was therefore neither utilized for normalization nor for clustering analysis. miRNAseq was performed to a median of 17M total reads (range 8-22M).From SNP array analysis, all 13 analyzed tumors displayed substantial degrees of aneuploidy typified by areas of complex genomic rearrangements suggestive of chromoanagenesis events sGiven the evidence of recurrent structural alterations in the vicinity of the chromosome 19 microRNA cluster as well as evidence in another tumor type that translocations to this area can lead to significant C19MC overexpression, we manually inspected mapped whole transcriptome sequencing in this region. Similar to observations in embryonal tumors with multilayer rosettes (EMTR) affected by C19MC fusion [ZIM2/PEG3 and C19MC, Chr19:54,178,123::Chr19:57,345,748 (Hg19 genome build) . Strikine build) suggestiSeven of the UESL tumors with sufficient materials underwent small RNA sequencing as well as an adjacent normal liver tissue from one patient and a Hepatocellular carcinoma cell line for comparison. All seven of the evaluated UESL tumors displayed substantial overexpression of the C19MC microRNAs to levels 100\u201310,000 fold higher than observed in either of the comparators . This leAmong the 46 individual C19MC miRNAs, 24 had information of both mature 3p and 5p miRNA expression levels and were classified based on the dominant expression of 3p or 5p miRNAs. Eight miRNAs had dominant expression of 3p over 5p, thirteen miRNAs had dominant expression of 5p over 3p, and three miRNAs had nearly equal amount of 5p and 3p mature miRNAs . NotablyTo further understand the expression and function of C19MC miRNAs in the context of other miRNAs, we performed correlation analysis of all expressed miRNAs in the 7 sequenced UESL samples. In UESL, the majority of the C19MC microRNAs (set-1) formed a tightly correlated expression pattern, suggesting shared expressional regulation of the majority of the cluster . NotablyAdditionally, a smaller subset of C19MC miRNAs (Set-2) formed a distinct correlatively co-expressed cluster, which was negatively correlated to a separate large set of TP53 and KRAS-regulatory miRNAs (Set-2) . Micro-RIn summary, C19MC microRNAs are substantially overexpressed in UESL and exhibit a strong negative correlation to K-Ras and TP53 regulatory miRNAs.TP53, was statistically recurrently mutated with somatic mutation identified in 5 of 7 patients with matched sequencing data with both mutational and CNV data harbored either TP53 mutation or copy number loss . From the matched sequencing cohort, we noted several additional possibly pathogenic somatic mutations including singleton mutations in 3 different JAK-STAT pathway genes, though none other than TP53 that were recurrent at the gene level , we observed a median of 39 somatic coding mutations per sample (range: 21\u201377), placing UES on the low end of the mutational burden spectrum across cancer types, similar to other pediatric malignancies. Only a single gene, ing data . To furtber loss . One addne level . Given rATF4, HIST1H1E, HIFX, and JUND response in placental physiology. Concordantly, we find differentiation and viral response gene sets to be the most highly enriched in UESL. Considering the role of C19MC in antiviral response and our function. FurtherTP53 loss/mutation, further study is warranted to examine the oncologic consequences of C19MC overexpression. Given the central importance of C19MC in UESL development, UESL would be well placed as a model system for further studies on this oncomir with potentially wide applicability of findings across cancer types.Given the increasing interest in C19MC as an oncomir cluster across multiple cancer types and the notable relationship between C19MC overexpression and Tissue samples were obtained from the Children\u2019s Oncology Group (COG) rare tumor repository. Tumors were classified as an embryonal sarcoma by a sarcoma pathologist at the host institution using standing histological techniques. We initially received extracted nucleic acids or representative tissue samples from 19 patients diagnosed with UESL, including 10 matched tumor-normal pairs and 9 patients with tumor only samples. Matched normal samples were whole blood for 9 patients and adjacent liver tissue for 1 patient. Fresh frozen tissue was utilized for 18 of the tumors, with formalin fixed tissue for the remaining 1 sample. Accompanying patient demographic data and deidentified pathological reports were also reviewed. For a subset of patients with quality concerns from the materials received, we performed post-hoc secondary pathological analysis of histological images from 5 tumor samples of concern. Upon our review, images from 4 of the 5 tumors lacked evidence of any viable tumor, displaying entirely necrosis and/or normal liver. The remaining tumor displayed a mix of necrosis (approximately 30%) with viable sarcoma cells. Given these findings, we excluded these 5 samples from analysis and proceeded with the remaining 14.All specimens for sequencing were obtained from patients with appropriate consent from their local institutional review board in accordance with the Children\u2019s Oncology Group and the National Cancer Institute. DNA/RNA were extracted from qualifying tumor samples (fresh frozen/FFPE) and matched blood using either AllPrep DNA/RNA Mini Kit (Qiagen #80204) or AllPrep DNA/RNA FFPE Kit (Qiagen #80234) or miRNeasy FFPE Kit (Qiagen #217504) or miRNeasy Mini Kit (Qiagen #217004). The RNA samples were subjected to on-column RNAse free DNAse (Qiagen #79254) digestion as per manufacturer\u2019s protocol. The nucleic acids were quantified using Qubit, and subjected to quality control using Bioanalyzer before proceeding to sequencing.Molecular studies included whole exome sequencing on all tumors and matched normal where available and with nucleic acids of sufficient quantity and quality. Two hundred ng of DNA was used as input into the Agilent SureSelect XT Clinical Research Exome kit, which includes the exon targets of Agilent\u2019s v5 whole-exome kit, with increased coverage at 5000 disease-associated targets. For each DNA sample, a genomic DNA library was constructed according to the manufacturer\u2019s protocol, and the size and quality of the library was evaluated using the Agilent BioAnalyzer. An equimolar amount of library DNA was used for a whole-exome enrichment using the Agilent capture baits and after quantitative PCR library quantitation and QC analysis on the BioAnalyzer. Paired-end 76bp sequencing was performed on Illumina NextSeq 500 instrument. The Burrows-Wheeler Aligner was used to align sequence reads to the reference genome hg19. The GenoRNA samples were processed for RNA-sequencing using the NuGen Ovation Human FFPE RNA-Seq Multiplex System . Briefly, 100 ng of RNA was used to generate cDNA and a strand-specific library following the manufacturer\u2019s protocol. Quality control steps including BioAnalyzer library assessment and quantitative RT-PCR for library quantification were performed. The libraries were then sequenced the Illumina NextSeq 500 v2 sequencer with 75-base paired-end runs in order to generate approximately 85M million read pairs per sample. Raw sequence reads were mapped to reference genome (hg19) using TopHat2.0. Fusion analysis was done using TopHat 2.0 and DeFuse 0.6. Expression FPKM results were obtained at both gene and transcript level using CuffLinks 2.1. Averages were taken for all TPM normalized genes in UESL samples and the 682 genes that show average expression >50 were subjected to EnrichR analysis to identSNP arrays were performed on tumor specimens with sufficient DNA quantity and quality using the CytoScan HD array from 250 ng of DNA according to the manufacturer\u2019s protocol (Thermo Fisher). Copy number state and allelic ratio were assessed utilizing the Chromosome analysis suite v3.3.0.139 (Thermo Fisher) and confirmed by manual inspection.Targeted sequencing of the C19MC breakpoint region was performed utilizing a custom targeted DNA panel designed to target the region from chr19:54139995-chr19:54220005 using hg19 coordinates. The design was predicted to cover 96.3% of the target capture region. Tumor DNA underwent library preparation and target enrichment using single primer extension technology according to the manufacturer\u2019s protocol and then subjected to 150bp paired-end sequencing using the Illumina NextSeq 500 platform. Sequencing reads were mapped to hg19 genome using the BWA algorithm. Fusion detection tools including Breakdancer, Manta5, and InFFor a candidate fusion in tumor from patient ID: PATWXD, primers were designed to span the predicted breakpoint. The primer combination that map to C19MC (5\u2019- ATGGTCAGCCTGGGCAGGGTAGC-3\u2019) and ZIM2/PEG3 gene (5\u2019- TCCTTTGCCCGAGGGCTCATGTTG-3\u2019) were used to amplify the fusion sequence, purified using GFX columns and subjected to Sanger sequencing using the primers listed above in addition to another set of nested primers which are designed based on IGV. The nested primers are for C19MC, 5\u2019- ACCTTGGTCAATATGGCGAAACCC-3\u2019 and 5\u2019- GATGTTCCTGTTCCCACAATTCTGG-3\u2019. Using the Sanger sequence, the breakpoint was mapped based on hg19 coordinates.Fresh frozen UESL and FFPE normal samples were subjected to RNA isolation using miRNeasy mini kit (Qiagen # 217004) and the samples with quality RNAs were subjected to small-RNA-sequencing using the QIAseq miRNA Library Kit according to the manufacturer\u2019s protocol (Qiagen). An average of 17M reads per sample were generated on an Illumina NextSeq 500 instrument using 75 bp single reads, and the raw data were analyzed using the QIAseq miRNA Primary Data Analysis pipeline. Unique molecular identifiers (UMIs) were counted, and miRNA sequences were mapped and counted using the Qiagen software. miRNAs that had cumulative expression above 100 in 7 UESL samples were log transformed and subjected to correlation analysis using R using package \u2018corrplot\u2019 0.84 (was built under R version 3.4.4). The scripts used were, > cor(FileName); > mat <- cor(FileName); > col1 <- colorRampPalette); > corrplot). The positively correlated C19MC miRNA cluster and corresponding negatively correlated miRNA clusters were subjected corrplot using the scripts above with an additional script for significance: > res1 <- cor.mtest; > corrplot, p.mat = res1$p, insig = \"blank\"). Therefore, white or blank represents low or no significant correlation.UESL miRNA-seq data was used to examine 3p versus 5p mature miRNA expression by calculation of cumulative expression of individual C19MC miRNAs in the sequenced UESL samples. The cumulative 5p and 3p data for C19MC miRNAs were used to get the 5p to 3p ratio and the miRNAs were classified into 3p dominant if the ratio is \u22640.5, neutral if the ratio is close to 1 and 5p dominant if the ratio is \u22651.5.S1 FigFor each tumor, SNP array data are plotted including log2 ratio (top) and allele difference (bottom) to provide an overview of genome-wide CNV. Figures were generated using Chromosome Analysis Suite.(TIF)Click here for additional data file.S2 FigTP53 mutations are the genomic hallmarks of UESL.C19MC overexpression due to chromosomal structural event and (TIF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file."} +{"text": "Fmr1 knockout (KO) mice, a model of FXS. The amounts of mitochondrial transcription factor NRF-1, ATP synthases ATP5A and ATPB, and the mitochondrial membrane protein VDAC1 in EVs were reduced in cerebral cortex samples and astrocytes from Fmr1 KO mice. These reductions correspond to decreased mitochondrial biogenesis and transcriptional activities in Fmr1 KO brain, along with decreased mitochondrial membrane potential (MMP) with abnormal localization of vimentin intermediate filament (VIF) in Fmr1 KO astrocytes. Our results suggest that mitochondrial dysfunction in astrocytes is associated with the pathogenesis of FXS and can be monitored by depletion of components in EVs. These findings may improve the ability to diagnose developmental diseases associated with mitochondrial dysfunction, such as FXS and autism spectrum disorders (ASD).Mitochondrial dysfunction contributes to neurodegenerative diseases and developmental disorders such as Fragile X syndrome (FXS). The cross-talk between mitochondria and extracellular vesicles (EVs) suggests that EVs may transfer mitochondrial components as intermediators for intracellular communication under physiological and pathological conditions. In the present study, the ability of EVs to transfer mitochondrial components and their role in mitochondrial dysfunction in astrocytes were examined in the brains of FMR1 gene, which encodes fragile X mental retardation protein (FMRP) hydrazinylyidene) propanedinitrile) for 30 min were used as a negative control. After adding buffer B, astrocytes were imaged using an A1RSi confocal microscope system (Nikon) with a 40\u00d7 lens objective. The MMP was assessed by quantifying the ratio of the intensity of orange fluorescence (emission wavelength 570 nm) to green fluorescence (emission wavelength 530 nm).The MMPs of astrocytes (DIV27) were determined by a MITO-IDCultured primary cortical astrocytes (DIV27) were briefly washed with phosphate-buffered saline (PBS) and then fixed with 4% paraformaldehyde in PBS. The cells were permeabilized with 0.25% Triton X-100 in PBS and incubated with PBS containing 10% donkey serum for 1 h. The cells were then incubated with primary antibodies against vimentin (1:500) and GFAP (1:300) for 1 h at room temperature, followed by washing and incubation with Alexa Fluor 488- or Alexa Fluor 594- conjugated anti-IgG for 1 h. After three washes (5 min each) using PBS, cells were mounted in Vectashield mounting medium with DAPI for fluorescence analysis. The images were acquired using an A1RSi confocal microscope system equipped with a 40\u00d7 lens objective.t-test. All values are expressed as the mean \u00b1 SD. Statistical significance was evaluated by an unpaired two-tailed"} +{"text": "Bertiella studeri tapeworms infecting children in Sri Lanka. Our findings can be used to identify multiple species of Bertiella tapeworms that can infect human hosts in the Old World.We provide a detailed molecular and phylogenetic description of Bertiella, which has 29 known tapeworm species, belongs to the subfamily Anoplocephalinae of the Anoplocephalidae family .This study provides the molecular analysis of the We conducted a retrospective study using tapeworm proglottids Figure 1https://www.thermofisher.com). We amplified 2 mitochondrial markers, nicotinamide adenine dinucleotide hydrogenase subunit 1 gene (NAD1) and cytochrome c oxidase subunit 1 gene (COX1), and 3 nuclear ribosomal markers, the second internal transcribed spacer region (ITS2), 28S large subunit ribosomal region (28S), and 18S rRNA gene (18S), using the specified primers and PCR conditions . The COX1 sequence similarity search identified 95.10% match with Bertiella species (GenBank accession no. JQ771106); COX1 analysis identified 2 clades in the Bertiella monophyletic group from Callicebus oenanthe monkeys were separated from the Sri Lanka clade and 100% similarity with Bertiella species (GenBank accession no. JQ771096). All the B. studeri sequences from Asia are in 1 clade. The second clade included 3 sequences from Pan troglodytes chimpanzee in Kenya, 1 from a human infection acquired in Equatorial Guinea, and 1 from a human host in Brazil (Bertiella from humans and nonhuman primates (Bertiella species (GenBank accession no. KJ888951). Furthermore, we identified a single-nucleotide polymorphism in 28S rRNA region (T to C) between the samples from Sri Lanka that suggest genetic diversity (B. studeri obtained from Macaca fascicularis macaque formed a single clade (B. studeri (GenBank accession no. GU323706). Furthermore, 18S rRNA region in the Sri Lanka samples have a single-nucleotide polymorphism (T to C) with the Bertiella sequence from M. fascicularis and gray langur (Presbytis entellus), that inhabit this region , and 3 nuclear ribosomal markers and submitted them to GenBank (B. studeri tapeworms. Our data may also be used to assist in elucidating if multiple species of Bertiella sp. tapeworms infect human hosts in the Old World.Unavailability of molecular data for GenBank . The molBertiella studeri infection in children, Sri Lanka.Additional information about"} +{"text": "Blood samples were collected from participants in the REGARDS study on two separate occasions. No participants in the Caregiving Transitions Study were caregivers at the first blood draw, but 251 became caregivers before the second blood draw 9 years later. These caregivers were matched with 251 noncaregiving controls. Six circulating biomarkers of inflammation and a measure of cellular aging (leukocyte telomere length) were assessed at both blood draws. All biomarkers except CRP showed overall aging effects (ps < 0.001). Caregivers had a small but significantly greater increase in TNFR1 levels (p = 0.03) than controls, but no significant differential changes were found on the other 5 inflammatory biomarkers or on telomere length. Preliminary findings from latent variable models indicated good model fit and found caregivers to be 0.27 SDs lower than controls on a latent construct of inhibitory, regulatory feedback of systemic inflammation (p = 0.03)."} +{"text": "According to Temporal Self-Regulation Theory , adherence motivation can be driven by both positive and negative emotional reactions in which adherence is viewed in relation to gains versus losses. In a sample of 100 older adults (ages 64+), we explored participant-provided feedback related to intervention and game elements participants perceived would increase their adherence across five domains: (1) ability to adjust difficulty; (2) ability to change game design (from the preprogrammed American Western theme); (3) cognitive performance feedback; (4) ability to unlock extra game features ; and (5) other elements. Ranked in order of frequency of endorsement, 53% endorsed: cognitive performance feedback; 47%: ability to adjust difficulty; 28%: ability to unlock new features; 27%: the \u2018Other\u2019 option; and 20% endorsed changing the game\u2019s theme. This work has implications for models of adherence, specifically, the role that expectations of later cognitive feedback might play."} +{"text": "Candida albicans to identify 13 cellular events occurring during neutrophil extracellular trap formation (NETosis) and mouse and human blood-derived polymorphonuclear neutrophils (PMNs) stimulated with ionomycin, lipopolysaccharide (LPS), or In their letter to the editor, Liu challenges our conclusion . Liu use1)C. albicans. c) Sollberger et al. (Published papers support our findings and contradict those of Liu: a) Neubert et al. showed ar et al. showed a2)Liu\u2019s flow cytometry analysis requires cells in suspension, while ours and other studies report actin disassembly during NETosis of adherent cells , 3, 5. N3)Liu defines time 0 as the addition of PMA stimulant, while we define time 0 as the first event visible by differential interference contrast microscopy (microvesicle shedding) after addition of stimulant marking entry into NETosis. We found that the time from stimulation to microvesicle shedding was highly variable, occurring between 0 min and 211 min (mean = 15\u00b133) after ionomycin addition, and not all stimulated cells entered or completed NETosis (4)We verified actin disassembly using three different neutrophil types and two different stimulants, and actin filament depolymerization is visualized using four different actin probes [GFP-actin , F-tractThe points below support our published contention that actin disassembly is an early event in NETosis."} +{"text": "Recent landmark studies have shown a survival benefit from using genomic profiling to guide targeted personalised therapy in men with metastatic castrate refractory prostate cancer (mCRPC) . The poiPatients referred to a standard NHS diagnostic service between Jan 2019 -Dec 2021 were screened using the pre-biopsy MRI report and presenting PSA . Eligibl62 men were recruited of whom 52 (84%) had cancer samples sufficient and met quality for sequencing. Cohort details are in Focusing on the 37 men with CPG4-5/metastatic disease, all but two had combined androgen deprivation therapy (ADT) and radiotherapy or systemic therapy alone (ADT and chemotherapy). Follow up was available for a median of 26 months (range 15-35). In this CPG4-5/metastatic sub-group; overall there were 12/37 (32%) progression events; 9 new or worsening metastases and 4 who progressed and died of prostate cancer. Three others developed rapid biochemical relapse after first-line treatment. Of the 5 men with DDR pathway mutations, 2/5 progressed; one with a BRCA2 mutation died from disease and another developed new metastasis. Interestingly 1 of the men with ATM loss of uncertain significance, relapsed rapidly after surgery. Of the 6 men with any Pi3K-Akt mutation there was 1 death from prostate cancer and 1 progression despite treatment. Combining men with either a DDR or a Pi3K pathogenic pathway mutation 4/11 (36%) progressed and/or died of disease (1 man had both BRCA2 and PI3K mutation). In men without any reported DDR or PI3K mutations (pathogenic or uncertain), 5/22 (23%) progressed in the follow-up period.(i)Targeting men with high PSA and/or MRI defined high probability lesions(ii)Histology identification of tissue to be profiled at routine pathology assessment(iii)Assign prognostic/metastatic category at MDT and determine likely outcomes vs competing morbidity for case selection(iv)Sequencing of those men with a high likelihood of progression or cancer mortality \u2013: CPG4-5/metastatic diseaseIt is now clear that specific genomic signatures add value in management decisions and impact disease trajectory and survival outcomes . FallingWe have shown that these steps are possible in a routine NHS diagnostic pathway and offers a rationale way to maximise the detection of potentially actionable mutations while avoiding over-investigation. Using this method DDR and PI3K mutations were detected in 11% and 19% in CPG4-5/metastatic disease and these men seemed to be over-represented in cases that progressed over the short term .a priori knowledge of BRCA status may allow modifications in earlier stages of management and future proof for evolutions in genomic testing . BRCA2 status for instance is already an independent predictor of poor survival in the Predict Prostate algorithm are now approved by NICE (as well as the European Medicines Agency and US FDA) . It coullgorithm .In summary we report a pragmatic method to optimise selection of men for molecular profiling of prostate cancer at the point of diagnosis. Working within standard diagnostic pathways our method could be adopted by any NHS (or other) unit without much resourcing. The upfront information could be used not only for future drug selection, but also influence decision making and personalised follow-up.Supplementary File"} +{"text": "Forkhead box A1 (FOXA1) encodes a pioneer factor that induces open chromatin conformation to allow the binding of other transcription factors. Through direct interactions with the Androgen Receptor (AR), FOXA1 helps to shape AR signaling that drives the growth and survival of normal prostate and prostate cancer cells. FOXA1 also possesses an AR-independent role of regulating epithelial-to-mesenchymal transition (EMT). In prostate cancer, mutations converge onto the coding sequence and cis-regulatory elements (CREs) of FOXA1, leading to functional alterations. In addition, FOXA1 activity in prostate cancer can be modulated post-translationally through various mechanisms such as LSD1-mediated protein demethylation. In this review, we describe the latest discoveries related to the function and regulation of FOXA1 in prostate cancer, pointing to their relevance to guide future clinical interventions.Prostate cancer is the most commonly diagnosed non-cutaneous cancers in North American men. While androgen deprivation has remained as the cornerstone of prostate cancer treatment, resistance ensues leading to lethal disease. Prostate cancer is the most commonly diagnosed non-cutaneous cancer in North American men cohort revealed FOXA1 mutation in 41% of localized prostate tumours, a rate significantly higher than Western cohorts into the proximity of a FOXA1 CRE termed FOXMIND and somatic SNVs can modulate FOXA1 genomic binding and transcription regulation. Genome-Wide Association Studies (GWAS) identified more than 160 risk loci to be associated with prostate cancer susceptibility can serve as a means to track minimal residual disease since normal tissues should not exhibit FOXA1 mutations. A targeted sequencing of 72 prostate cancer driver genes in the plasma cell-free DNA of patients with mCRPC revealed 37 somatic mutations mapped to FOXA1 3\u2032 UTR in 34/290 (12%) of the patients (Fig.\u00a0FOXA1 harboured the highest 3\u2032 UTR mutation rate among all 72 genes of interest with indels being the dominant type of alteration (Annala et al. FOXA1 mutations can be readily detected in liquid biopsy and can potentially serve as a prostate cancer-specific biomarker. Recent findings have shown the differential prevalence of FOXA1 mutations across prostate cancer stages, for instance the early emergence of Forkhead Wing2 coding mutations in localized disease and the enrichment of truncation mutations and SVs in metastatic diseases (Adams et al. FOXA1 can inform disease progression. Based on the varying function of FOXA1 mutants, the prognostic value of different FOXA1 mutations should be further evaluated in future studies. Through identifying inhibitors that antagonize specific FOXA1 mutants, this approach potentiates personalized treatment. With the constant cellular tumour cellular turnover releasing cell-free DNA, liquid biopsy for FOXA1 mutations permits longitudinal tracking for residual disease and early detection for recurrence.nts Fig.\u00a0. In factFOXA1 mRNA expression that are extensively perturbed through various modes of somatic mutations. In addition, post-translational modifications such as protein demethylation mediated by LSD1 are identified as additional regulations of FOXA1 activity. Deeper understanding of the somatic mutations and regulatory mechanisms may yield novel biomarkers and therapeutic strategies complementary to our existing standard of care with the ultimate goal of improving patient outcomes. With present day high-throughput methodologies and model systems that have been lacking in the past, there is promise in translating the past and recent biological findings regarding FOXA1 into clinical utility.Recent functional characterization of FOXA1 mutants shed light on their complex modes of action in driving prostate cancer where the function may be divergent depending on the residues affected. Overlaying the prostate cancer epigenome reveals a catalog of CREs critical for"} +{"text": "The Sumatran rhinoceros is critically endangered, with fewer than 100 individuals surviving across its current range. Accurate census estimates of the remaining populations are essential for development and implementation of conservation plans. In order to enable molecular censusing, we here develop microsatellite markers with amplicon sizes of short length, appropriate for non-invasive fecal sampling.A\u2009=\u20092.4; HO\u2009=\u20090.30). These were sufficient to distinguish among individuals (PID\u2009<\u20090.0001), and to distinguish among siblings (PID(sib)\u2009<\u20090.0001). Among rhinos in Indonesia, almost all markers were established as polymorphic and effective for genotyping DNA from fecal samples. Notably, the markers amplified and displayed microsatellite polymorphisms using DNA extracted from 11 fecal samples collected non-invasively from wild Sumatran rhinoceros. These microsatellite markers provide an important resource for a census and genetic studies of wild Sumatran rhinos.Due to limited sample quantity and potential lack of genome-wide diversity, Illumina sequence reads were generated from two Sumatran rhinoceros samples. Genomic screening identified reads with short tandem repeats and loci that were polymorphic within the dataset. Twenty-nine novel polymorphic microsatellite markers were characterized (The online version contains supplementary material available at 10.1186/s13104-021-05522-x. Dicerorhinus sumatrensis) has declined by more than 50% [In the past two decades the population of Sumatran rhinoceros identification of 29 novel short amplicon polymorphic Sumatran rhinoceros microsatellite markers, (b) characterization of variability of these markers using high quality DNA, (c) optimization of amplification success and verification of polymorphisms of these markers using DNA from dung, and (d) first field results from use of these markers on 11 dung samples from free-ranging Sumatran rhinoceros in Indonesia. Our results suggest that these new tools will be of value in the conservation and management of extant Sumatran rhinoceros.n\u2009=\u20096) from the San Diego Zoo Institute for Conservation Research (ICR) and the Royal Ontario Museum (ROM); blood samples from rhinos kept at the Cincinnati Zoo (n\u2009=\u20092) or at the Sumatran Rhino Sanctuary (SRS) within Way Kambas National Park (WK) in Sumatra (n\u2009=\u20093); fecal samples from captive rhinoceros at the Cincinnati Zoo (n\u2009=\u20092) and SRS (n\u2009=\u20093); and fecal samples (n\u2009=\u200911) from an unknown number of wild rhino individuals at the Burkit Barisan Selatan National Park (BBS) in Sumatra. Across these samples, there were paired blood-fecal samples for three of the rhinos (two at the Cincinnati Zoo and one at SRS).We used several types of Sumatran rhino samples Table : previoun\u2009=\u20092) using the Qiagen DNeasy Blood and Tissue Kit. At the Eijkman Institute for Molecular Biology (EIMB), DNA was extracted from the SRS rhino blood samples (n\u2009=\u20093) using a salting out procedure [n\u2009=\u20092), SRS (n\u2009=\u20093) and BBS (n\u2009=\u200911) using the QIAmp DNA Stool Kit (Qiagen) with a modified protocol [At the University of Illinois at Urbana-Champaign (UIUC), high quality DNA was extracted from the Cincinnati Zoo blood samples (rocedure . At bothprotocol . PCR with the IPCRESS program [Whole genome sequences (Illumina MiSeq v3) were generated from two high quality DNA samples from Sumatran rhinoceros Dsu-33 and Dsu-35, both originally from Sumatra Table . Reads f program , loci shCandidate primer pairs were evaluated at UIUC , expected heterozygosity (HE), and observed heterozygosity (HO). CERVUS v3.0.7 [PID and PID(sib) values for each marker, and cumulative PID and PID(sib) across multiple markers, using the equations of Waits et al. 2001 [PID and PID(sib) were calculated by multiplying PID and PID(sib) across loci. Using GENEPOP (https://genepop.curtin.edu.au/), significant linkage disequilibrium was not detected among loci. Genome scaffolds were identified for loci from rhino Dsu-33 and 167,849 reads (2.8%) from Dsu-35 contained microsatellite motifs. Our bioinformatics method identified 861 polymorphic di-, tri-, and tetra-nucleotide repeat loci within the two sequenced Sumatran rhinos of which 229 had suitable priming regions. Loci were then excluded if they displayed\u2009<\u20096 tandem repeats, had too broad a size range, showed evidence of being in repetitive elements, or closely matched sequences of human DNA .A)\u2009=\u20092.4; number of alleles ranged from two to four; average expected heterozygosity (HE)\u2009=\u20090.45; and average observed heterozygosity (HO)\u2009=\u20090.30. Overall, FIS\u2009=\u20090.44, likely due to population structure among geographically separated populations, or to inbreeding. Using the most informative markers, i.e., those with the lowest PID values, as few as seven loci could cumulatively distinguish individual identity (cumulative PID\u2009<\u20090.0001 [PID(sib), as few as 19 optimized loci could confidently distinguish individual identity among siblings (cumulative PID(sib)\u2009<\u20090.0001 [The above resulted in 55 polymorphic candidate loci for UIUC laboratory PCR optimization using six test samples fecal samples are sometimes difficult to distinguish in the field. The specificity of primer pairs was tested using Asian tapir DNA and human DNA (a conceivable contaminant). One or both of these species amplified for 12/29 markers, although their amplicon sizes were almost always substantially different from those of rhino (Additional file To further test their utility for a census using dung collected from wild Sumatran rhinos, the 13 randomly chosen new markers were tested on 11 fecal samples collected from an unknown number of wild rhinoceros in BBS (Table After species identity was established for 11 fecal samples from wild rhinos, the 13 randomly chosen markers were tested. For nine of the 13 markers, DNA from least nine of the 11 fecal samples amplified successfully and could be scored (Additional file Recent estimates suggest that fewer than 100 Sumatran rhinoceros individuals persist across Sumatra, with few in Borneo, although current census estimates have a large degree of uncertainty . Since aAs additional fecal samples are collected from wild rhinoceros, PCR conditions and marker choice may be further optimized for some primer pairs Table . AlthougAdditional file 1. Detailed methods, supplementary figures and tables."} +{"text": "The tako tsubo cardiomyopathy (TTC) is characterized by a transient contractile dysfunction after severe physical or emotional stress. Relevant coronary artery disease is absent. Clinically it mimics acute myocardial infarction (AMI). Therefore, the aim of this study was to characterize morphological differences between TTC and AMI using different MRI parameters.22 TTC and 35 AMI patients were examined within 48 hours upon admission. Ejection fraction (EF), myocardial mass (MM) enddiastolic (EDV) and endsystolic volumes (ESV) and regional contractility scores were computed on 10 contiguous CINE SSFP short axis slices. Myocardial edema (ME) was computed on T2 weighted TSE images on short axis orientations, late enhancement (LE) was evaluated on FLASH 3D GRE . Measurements are given as mean \u00b1 SE. Differences were computed using an ANOVA model, a p value =< 0.05 was cosidered significant.All AMI patients and none of the TTC patients included had LE. TTC patients had a significantly lower EF all other comparisons were therfore controlled for EF. TTC patients had significantly smaller ventricular volumes myocardial mass and edema . In contrast regional contractility was more affected in TTC with a significantly larger number of dysfunctional segments and a significantly higher wall motion score .In contrast to AMI, TTC patients show a marked regional dysfunction but no signs of early remodeling like ventricular enlargement and increased myocardial mass."} +{"text": "Arabidopsis. OsProDH encodes a putative proline dehydrogenase and is a single copy gene in rice. However, whether OsProDH plays roles in abiotic stress in rice remains unknown.Global warming threatens rice growth and reduces yields. Proline plays important roles in plant abiotic stress tolerance. Previous research demonstrated that engineering proline metabolism-related genes can enhance tolerance to freezing and salinity in OsProDH transcript contents were relatively higher in leaf blade and root tissues and the high temperature treatment repressed expression of OsProDH. The predicted OsProDH protein localized in mitochondria. Using the Oryza sativa ssp. japonica cultivar KY131, we generated OsProDH overexpression (OE) lines and knockout mutant lines using the CRISPR/Cas9 (CRI) system. Overexpression of OsProDH decreased proline content, while mutation of OsProDH increased proline content compared with that of KY131. The CRI and OE lines were respectively more resistant and sensitive to heat stress than KY131. Heat stress induced proline accumulation and mutation of OsProDH led to proline overproduction which reduced H2O2 accumulation in the seedlings.Quantitative RT-PCR analysis revealed that OsProDH negatively regulates thermotolerance in rice. Our study provides a strategy to improve heat tolerance in rice via manipulating proline metabolism. High temperature stress reduces plant growth and crop productivity, potentially resulting in widespread risk of food insecurity (Battisti and Naylor 1-pyrroline-5-carboxylate (P5C) which is subsequently converted into glutamate by P5CDH encoding proline dehydrogenase in Arabidopsis. The predicted pre-proteins AtProDH1 and AtProDH2 share 75% identical amino acids , a single copy gene encoding the putative proline dehydrogenase in rice. We cloned the coding sequence (CDS) and genomic DNA sequence of OsProDH by PCR method using the japonica variety Kongyu131 (KY131). We compared the CDS with genomic DNA and found four exons in OsProDH genomic DNA. The predicted pre-proteins of OsProDH has 454 amino acids and with the proline dehydrogenase domain located at residues 133\u2013436 by searching the NCBI Conserved Domain Database analysis revealed that ues Fig.\u00a0a. To exa35S::OsProDH-GFP vector was introduced into rice protoplasts. As AtProDH1 and AtProDH2 were all localized in mitochondria lines and knockout mutant lines using CRISPR/Cas9 (CRI) system under KY131 background. Two OE and CRI lines were chosen and characterized in detail for this study . However, under high temperature stress conditions, CRI and OE lines were respectively more resistant and sensitive to heat stress than KY131 staining to visually evaluate H2O2 accumulation in leaves. The brown precipitate indicative of H2O2 accumulation was generally not distributed in the leaves of both KY131 and transgenic lines prior to the heat treatment derived from the CRISPR-Cas9 system.Additional file 3: Table S1. Primers used in this study were listed."} +{"text": "Restoration of normal epicardial coronary flow in acute ST segment elevation myocardial infarction does not ensure adequate perfusion at the myocardial tissue level. In the era of primary PCI, patients with acute myocardial infarction treated successfully with PCI are discharged mostly within first week. There are no studies available regarding blood flow at the tissue level in these patients during effort.Assessment of stess and rest perfusion defects early after STEMI successfully treated with pPCI.61 patients with first anterior STEMI (57 \u00b1 10 yrs. 52 M) who underwent successful pPCI have been included into the study. CMR was performed on 1.5 T scanner between 5 and 10 days after pPCI. Myocardial perfusion was assessed at rest and in stress condition during infusion of adenosine during first-pass perfusion imaging. Microvascular obstruction regions (MVO) were assessed on early enhancement images acquired 1\u20132-minutes after stress perfusion. Delayed enhancement (DE) images were acquiered 15 minutes after Gd-DTPA. Transmurality of myocardial perfusion defecits at rest and in stress condition, MVO were evaluated using 5 point scale in 16 segments. DE was also evaluated in segment 17. The sum of scores were calculated for each variable. Scar size and MVO were additionally quantitatively analyzed using MASS software. The results were given in ml and in % of LV volume.Only in 2 patients there was no evidence of at least subendocardial stress perfusion deficit. Stress perfusion sum of scores discriminated patients with normal EF (mean 59 \u00b1 1%) and LV dysfunction . Median stress perfusion sum of scores was 15 points (ranged 0 to 37). Median rest perfusion scores was significantly lower . Median MVO and DE sum of scores were 3 points (ranged 0\u201333) and 25 (3\u201342) respectively. DE measured as a % of LV volume has discriminated patients with severe vs moderate LV dysfunction with a cut of point 40%. Regression model revealed that only DE and stress perfusion sum of scores have significantly contributed to the LV EF model at discharge.Despite TIMI 3 flow in coronary artery myocardial perfusion defects at the tissue level are very frequent. Only in patients with preserved LV function pharmacological stress have not induced or intensed perfusion deficits. All patients with STEMI anterior successfully treated with pPCI, even with mild LV dysfunction should avoid effort which could induce ischemia. The pathogenesis remains unknown. Further studies are needed."} +{"text": "Mucosal malignant melanoma of the head and neck is a rare diagnosis. The safety and efficacy of salvage neck dissection following carbon\u2013ion radiotherapy with concurrent chemotherapy are not well described, and carbon\u2013ion radiation protocols have not been fully developed. A 77 year old woman with crT0N1M0 mucosal melanoma of the head and neck achieved a complete response following initial treatment with carbon\u2013ion radiotherapy and concurrent chemotherapy. She was treated with salvage neck dissection for as a cervical lymph node metastasis 16 months after initial treatment. She experienced neither Clavien-Dindo Grade 3 or 4 postoperative complications nor subsequent recurrence of disease at 3 months following salvage neck dissection. Surgical specimens may be useful for future precision oncology based on the molecular biology of recurrence melanoma with poor prognosis. Mucosal malignant melanoma of the head and neck (MMMHN) is very rare, accounting for less than 1% of all melanomas . Salvage18F-fluorodeoxyglucose positron emission tomography with CT (18FDG-PET/CT) from the head to the thighs revealed abnormal uptake of the palate without any neck lesions. The diagnosis was stage of cT3N0M0 MMMHN, based on the eighth edition of the American Joint Committee on Cancer Staging Manual and magnetic resonance imaging (MRI) revealed a 26 \u00d7 10 mm oral tumor without bone invasion or neck lymph node metastasis. g Manual . The patg Manual . ConcurrM0 MMMHN .After considering radiotherapy and systemic immunotherapy, the patient was treated by en bloc salvage ND with preservation of the accessory nerve, internal jugular vein, and sternocleidomastoid muscle, as described by the Japan Neck Dissection Study Group . CefazolThe poor survival outcomes of MMMHN highlight the need more effective treatment ,2. IndeeAdditional data on postoperative complications of salvage surgery of local recurrence following CIR are available for several types of malignant tumors ,8,10. SaA patient with stage crT0N1M0 MMMHN who underwent salvage ND after CIR with concurrent chemotherapy experienced neither grade 3\u20134 of Clavien-Dindo postoperative complications nor recurrence at 3 months of follow-up. Molecular biology from surgical specimens of recurrence MMMHN may be useful for future precision oncology."} +{"text": "Dear Editor,GABAergic interneurons (GIs) play an essential inhibitory role in regulating neural circuitry and neurological activities. Approximately 54% GABAergic interneuron population originated from medial ganglionic eminence (MGE) at E13.5 in rodents is a subtype gene of LIM homeodomain family. During early development, Lhx6 is expressed in the MGE and plays an essential role in the migration of GIs from MGE to the cortex in mice , which is reasonable that cells in the MGE mantle area could enhance SHH signaling , we constructed LHX6 knockout hESC lines and differentiated into GIs. We constructed LHX6 knockout hPSC lines by using a donor-free paired gRNA-guided CRISPR/Cas9 strategy could promote GIs migration. LHX6 conditional overexpression human pluripotent stem cell (hPSCs) lines was constructed as our previous report mouse brain slices Fig.\u00a0A. After To more systematically test the role of LHX6 in migration, we used fused forebrain organoids to determine the GI migration from ventral to dorsal Fig.\u00a0H. Cerebrin vivo, we transplanted the 7-week hPSCs-derived MGE progenitors into the ventral forebrain of severe combined immunodeficiency (SCID) neonatal mice , while in rodent studies that the GABA neuron number is similar in In conclusion, our results demonstrated that LHX6 is an essential transcription factor for the migration of human GABA interneurons by using knockout and overexpression strategies in human pluripotent stem cells. This method let us dig LHX6 function in human neurodevelopment from a new perspective. It could also be an important guide for the application of cell therapy in related diseases.Supplementary material 1 (PDF 1016 kb)Below is the link to the electronic supplementary material."} +{"text": "Scythropus yasumatsui (Coleoptera: Curculionidae) was determined by using an Illumina platform. The circular genome was 16,472\u2009bp in length and contained 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), 13 protein-coding genes (PCGs), and one control region. The nucleotide composition was significantly biased with A\u2009+\u2009T contents of 74.49%. All PCGs were initiated with standard ATN (ATG/ATT) codons. While 10 PCGs were terminated with TAA, two PCGs were terminated with TAG (cytb and nad1), and nad5 was terminated with an incomplete stop codon TA. All tRNAs were predicted to contain typical cloverleaf secondary structures except trnS1. The phylogenetic analysis of the concatenated nucleotide sequences of 13 PCGs from 12 Curculionidae species was performed by using MrBayes 3.1.2. The results indicated that S. yasumatsui was more closely related to Naupactus xanthographus than to other species.The complete mitochondrial genome sequence of Scythropus yasumatsui, is native to the Northern region of China where it has become a major pest of Chinese jujube, Ziziphus jujuba Mill., and contained 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), 13 protein-coding genes (PCGs), and 1 control region (non-coding AT-rich region). The order and orientation of the mitochondrial genes were identical to typical mtDNA of beetles , and nad5 was terminated with an incomplete stop codon TA. All of the 22 tRNAs were predicted to contain typical cloverleaf secondary structures except the gene trnS1 (AGN), whose DHU arm was replaced by a simple loop. rrnL was 1315\u2009bp in length with A\u2009+\u2009T contents of 78.86%, and rrnS was 785\u2009bp in length with A\u2009+\u2009T contents of 78.22%.The nucleotide composition was significantly biased with A\u2009+\u2009T contents of 74.49%. All PCGs were initiated with typical ATN codons (eight with ATT and five with ATG). Whereas 10 PCGs were terminated with TAA, two PCGs were terminated with TAG were used for the phylogenetic analysis. For the 12 Curculionidae species, they came from eight subfamilies or tribes, including Dryophthorinae , Cryptorhynchinae (Eucryptorrhynchus brandti and Eucryptorrhynchus chinensis), Entiminae (Naupactus xanthographus), Curculioninae (Curculio davidi), Cyclominae (Aegorhinus superciliosus), Molytinae (Hylobitelus xiaoi), Platypodinae (Platypodinae sp.), and Scolytinae (Scolytinae sp.), and their complete mitochondrial sequences were obtained from GenBank of NCBI. Bayesian analyses were performed with MrBayes 3.1.2 (Ronquist and Huelsenbeck S. yasumatsui was more closely related to Naupactus xanthographus than to other species (S. yasumatsui and species from the subfamily Entiminae.To further understand the mitogenome of species , which m"} +{"text": "Insulin resistance (IR) precedes the development of type 2 diabetes (T2D) and increases cardiovascular disease risk. Although genome wide association studies (GWAS) have uncovered new loci associated with T2D, their contribution to explain the mechanisms leading to decreased insulin sensitivity has been very limited. Thus, new approaches are necessary to explore the genetic architecture of insulin resistance. To that end, we generated an iPSC library across the spectrum of insulin sensitivity in humans. RNA-seq based analysis of 310 induced pluripotent stem cell (iPSC) clones derived from 100 individuals allowed us to identify differentially expressed genes between insulin resistant and sensitive iPSC lines. Analysis of the co-expression architecture uncovered several insulin sensitivity-relevant gene sub-networks, and predictive network modeling identified a set of key driver genes that regulate these co-expression modules. Functional validation in human adipocytes and skeletal muscle cells (SKMCs) confirmed the relevance of the key driver candidate genes for insulin responsiveness. Insulin resistance is characterized by a defective response (\u201cresistance\u201d) to normal insulin concentrations to uptake the glucose present in the blood, and is the underlying condition that leads to type 2 diabetes (T2D) and increases the risk of cardiovascular disease. It is estimated that 25\u201333% of the US population are insulin resistant enough to be at risk of serious clinical consequences. For more than a decade, large population studies have tried to discover the genes that participate in the development of insulin resistance, but without much success. It is now increasingly clear that the complex genetic nature of insulin resistance requires novel approaches centered in patient specific cellular models. To fill this gap, we have generated an induced pluripotent stem cell (iPSC) library from individuals with accurate measurements of insulin sensitivity, and performed gene expression and key driver analyses. Our work demonstrates that iPSCs can be used as a revolutionary technology to model insulin resistance and to discover key genetic drivers. Moreover, they can develop our basic knowledge of the disease, and are ultimately expected to increase the therapeutic targets to treat insulin resistance and type 2 diabetes. Insulin resistance is necessary for the development of the metabolic syndrome and type 2 diabetes (T2D), and is a major risk factor for cardiovascular disease , which tTo fill this gap, we sought to take advantage of a large library of induced pluripotent stem cells (iPSCs) derived from individuals across the spectrum of insulin sensitivity who have also undergone GWAS genotyping ,8. We haThese results prompted us to more specifically analyze the gene expression patterns and networks associated with the insulin sensitivity status of the iPSC donors. For complex conditions like insulin resistance with polygenic susceptibility, systems biology and network modeling, integrating multiscale-omics data like genetic and transcriptomic data, provide a useful context in which to interpret associations between genes and functional variation or disease states \u201313. Ther.In summary, we performed differential expression analyses between insulin resistant (IR) and insulin sensitive (IS) iPSCs, built co-expression networks to systematically organize the data into coherent modules enriched for insulin sensitivity associated functions and implemented key driver analyses to identify genes that control and regulate critical aspects of the IR and IS networks. Finally, we empirically validated the constructed networks in iPSCs and the prioritized key drivers through insulin responsiveness associated functional assays in human adipocytes and skeletal muscle cells (SKMCs)S1 Table). We generated three to seven iPSC lines from each individual, with no apparent differences in the reprogramming efficiency between IR and IS cells . The complete pipeline for iPSC generation and quality control has been previously described glucose for 10 minutes at room temperature. The cells were then washed with PBS, harvested in 300uL of M-PER lysis buffer (ThermoFisher), and then added to scintillation vials containing 4.75mL of scintillation fluid (Perk Elmer). Radioactive counts were determined with a scintillation counter (Model ID: Beckman LS6500). Excess samples were subjected to BCA assay for protein quantification and normalization of radioactive counts. All samples were represented as fold change compared to the unstimulated (no insulin) condition.Differentiated SGBS or HMCL-7304 cells cells were pretreated for 24 hours with different concentrations of the chemical inhibitors for HMGCR (atorvastatin), FDPS , and SQLE (terbinafine). After the preincubation, the cells were starved for 2 hours prior to 30 minute of 100nM insulin stimulation at 37 degrees. Following stimulation, cells were incubated with Krebs-Ringer bicarbonate-HEPES (KRBH) buffer Click here for additional data file.S2 Fig(TIF)Click here for additional data file.S3 Fig(TIF)Click here for additional data file.S4 Fig(TIF)Click here for additional data file.S5 Fig(TIF)Click here for additional data file.S6 Fig(TIF)Click here for additional data file.S7 Fig(TIF)Click here for additional data file.S8 Fig(TIF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLS)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLS)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S7 Table(XLS)Click here for additional data file.S8 Table(XLSX)Click here for additional data file."} +{"text": "Wuchereria bancrofti and Brugia spp. that istargeted for elimination by mass drug administration (MDA)1. In 2018, we published results from a clinicaltrial showing a single co-administered dose of ivermectin/diethylcarbamazine/albendazole(IDA) cleared microfilaremia in 55 of 57 individuals with bancroftian filariasis forthree years after treatment2. Thisresult was far superior to that obtained with the prior standard two-drugdiethylcarbamazine/albendazole regimen. Consequently WHO modified their guidelines torecommend IDA therapy for filariasis elimination outside of sub-Saharan Africa inregions that have not started or delivered fewer than four annual rounds ofdiethylcarbamazine/albendazole or have not met thresholds for transmissioninterruption3. Merck Inc.expanded its ivermectin donation by 100 million additional doses annually to helpfacilitate this change, with 68 million people expected to receive IDA next year.Lymphatic filariasis is a neglected tropical disease caused by the nematode parasites2, suggesting thatIDA sterilized adult worms without killing all of them. Since the estimated reproductivelifespan of filarial adult worms is five years5, it was possible that remaining worms might recover and startproducing microfilaria. To investigate whether IDA had sterilized adult worms, wereexamined 36 individuals approximately 5 years after single dose IDA treatment using the same parasitological methodsdescribed in the original study2.None of these individuals had received any subsequent treatment for lymphaticfilariasis. 35 of 36 individuals had zero microfilaria in 2 ml of venous night blood(One limitation of IDA treatment was that most individuals did not completely clearcirculating filarial parasite antigenht blood. One per"} +{"text": "Objective: Determine illness perceptions associated most frequently with measures of type-2 diabetes mellitus (T2DM) maintenance. We measured illness perceptions using the Illness Perception Questionnaire (IPQ) and variants (IPQ-Revised and Brief IPQ) Design: Review of literature from publication of IPQ to September 2020. Searched for articles utilizing IPQ but no other models of illness perception and studying T2DM Main Outcome Measures: Glucose control (measured by HbA1c levels), adherence to medications, and adherence to diet, exercise, and other lifestyle recommendations Results: Symptom attribution and fear of consequences are frequently associated with worse T2DM management and sense of control of illness progression and positive emotional valence are frequently associated with better T2DM management. Other subscales have less frequent but generally positive associations with the exceptions of recurring thoughts about T2DM duration, which had a negative association with management, and understanding the causes of T2DM, which had no associations at all. Other reviews found similar associations and highlighted a need for more general T2DM education. Conclusion: Future T2DM management interventions should promote sense of control over illness progression and positive emotional valence and provide education regarding symptoms to expect. Interventions should also consider managing patient fear, which has many associations with worse management."} +{"text": "This cohort study evaluates the outcomes associated with deferred vs expedited aortic valve replacement in patients with severe aortic stenosis during the COVID-19 pandemic. Between March 20 and April 26, 2020, the Federal Council of Switzerland banned elective procedures in all hospitals in Switzerland.Balancing the risk of deferral of treatment of symptomatic severe aortic stenosis vs expedited aortic valve replacement (AVR) during the time of health care restrictions requires careful clinical judgement. The aim of the present study was to prospectively evaluate outcomes in a cohort of patients with symptomatic severe aortic stenosis who received deferred vs expedited AVR based on prespecified criteria during the COVID-19 pandemic.2 This study was approved by Ethics Committee Bern, Switzerland. All patients provided written or oral informed consent for participation in this study. This study followed the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) reporting guideline.In this cohort study, the Aortic Stenosis Defer (AS DEFER) study, all patients with symptomatic severe aortic stenosis referred for AVR between March 20 and April 26, 2020, were consecutively included. Definitions, data collection, end point ascertainment, statistical analysis, and study flow are detailed in the eAppendix in the 2 or less, a transvalvular mean gradient of at least 60 mm Hg, cardiac decompensation during the previous 3 months, or exercise intolerance with clinical symptoms on minimal exertion were allocated to expedited AVR. Patients with an aortic valve area of 1.0 cm2 or less and greater than 0.6 cm2 and stable symptoms were allocated to deferred AVR. The primary end point was a composite of all-cause mortality, disabling and nondisabling stroke, and unplanned hospitalization for valve-related symptoms or worsening heart failure by intention to treat. A 2-sided P\u2009<\u2009.05 was considered statistically significant. Statistical analyses were performed using SAS software, version 9.4 (SAS Institute).Patients with critical aortic stenosis defined by an aortic valve area of 0.6 cmBetween March 20 and April 26, 2020, a total of 71 patients with symptomatic severe aortic stenosis were prospectively enrolled in the study. According to the prespecified algorithm, 25 patients (35.2%) were allocated to expedited AVR and 46 patients (64.8%) were allocated to deferred AVR. Baseline characteristics are summarized in the P\u2009=\u2009.08) (P\u2009=\u2009.02). Patients allocated to deferred AVR who required hospitalization for valve-related symptoms or worsening heart failure more commonly had multivalvular disease . Seven patients (15.2%) hospitalized for valve-related symptoms or worsening heart failure crossed over to expedited transcatheter AVR (n\u2009=\u20094) or surgical AVR (n\u2009=\u20093) within a mean (SD) of 17 (11) days after interdisciplinary allocation of the treatment strategy. One patient allocated to expedited transcatheter AVR experienced a periprocedural nondisabling stroke; none of the patients died. Compared with patients with no event, patients who experienced a primary outcome\u2013relevant event had a similar delay between confirmation of diagnosis and referral for AVR and comparable rates of New York Heart Association functional class of 3 or more (indicating marked symptoms during daily activity) at baseline .At a mean (SD) duration of follow-up of 31 (11) days after treatment strategy allocation, the primary composite end point occurred in 9 patients (19.6%) in the deferred treatment arm and in 1 patient (4.0%) in the expedited treatment arm (log rank P\u2009=\u2009.08) . Hospita3 Deferral of AVR in patients with symptomatic severe aortic stenosis was associated with an increased risk of hospitalization for valve-related symptoms or worsening heart failure. Patients with symptomatic severe aortic stenosis in combination with relevant multivalvular disease may particularly benefit from expedited AVR. Study limitations include the low patient number owing to the short duration of the ban of elective procedures and the locally modest numbers of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections, which did not exceed available health care resources.The algorithm used in this study to allocate the treatment strategy was similar to the triage recommendation from the American College of Cardiology and Society for Cardiovascular Angiography & Interventions consensus statement issued after initiation of patient recruitment for the study."} +{"text": "HOTAIR was proposed to regulate either HoxD cluster genes in trans or HoxC cluster genes in cis, a mechanism that remains unclear. We have identified a 32-nucleotide conserved noncoding element (CNE) as HOTAIR ancient sequence that likely originated at the root of vertebrate. The second round of whole-genome duplication resulted in one copy of the CNE within HOTAIR and another copy embedded in noncoding transcript of HOXD11. Paralogous CNEs underwent compensatory mutations, exhibit sequence complementarity with respect to transcripts directionality, and have high affinity in\u00a0vitro. The HOTAIR CNE resembled a poised enhancer in stem cells and an active enhancer in HOTAIR-expressing cells. HOTAIR expression is positively correlated with HOXC11 in cis and negatively correlated with HOXD11 in trans. We propose a dual modality of HOTAIR regulation where transcription of HOTAIR and its embedded enhancer regulates HOXC11 in cis and sequence complementarity between paralogous CNEs suggests HOXD11 regulation in trans. \u2022Two conserved noncoding elements (CNEs) overlap HOTAIR and its paralog on HOXD cluster\u2022cis and trans effects of HOTAIRParalog on HOXD cluster may resolve controversy over \u2022cis HOX genes but negatively with trans clusterCNEs are positively coregulated with \u2022Transcribed CNEs with coevolved complementarity suggest hybridization-based function Biological Sciences; Molecular Biology; Molecular Mechanism of Gene Regulation Only a ulation) . Althougon) in trans by recruiting the Polycomb Repressive Complex 2 (PRC2) in mouse showed limited impact on gene expression and H3K27me3 levels at Hoxd genes cluster genes (Hotair deletion (cis-regulation of HOTAIR is mediated via an unannotated enhancer element within its gene body or through transcription of the HOTAIR promoter remains unknown. Different regulatory mechanisms (cis versus trans) might be explained by tissue origin and changes in developmental stages in distinct genetic backgrounds . However2 (PRC2) and PRC2pression . Deletioxd genes . Specifi Hoxd13) . These oer genes . This stdeletion . Whetherkgrounds . As suchgulation . As the HOTAIR regulates HOXC cluster genes in cis (trans (cis and/or trans interactions. We have identified and characterized a 32-nucleotide conserved noncoding element (CNE) as the HOTAIR ancestral sequence, which is shared by both paralogous loci in HoxC and HoxD clusters, presenting itself in an inverted syntenic position. Strikingly, the paralogous CNEs underwent compensatory mutations during vertebrate evolution, which exhibit sequence complementarity dependent on the transcript orientation. Also, the CNEs have high interaction propensity revealed by microscale thermophoresis (MST). These observations suggest direct hybridization between the two noncoding transcripts. HOTAIR CNE represents either an active or poised enhancer in different cellular contexts. Its expression is positively correlated with HOXC11, whereas negatively correlated with HOXD11, suggesting dual modality of HOTAIR CNE in cis and trans.To address whether s in cis or HOXD s . As HOTAerlooked . To thiserlooked and idenerlooked A that iserlooked A and S1Berlooked A or HoxD cluster (HoxD11 and HoxD12 (reported target genes of HOTAIR in trans) in the HoxD cluster and between HoxC11 and HoxC12 (reported target genes of HOTAIR in cis) in the HoxC cluster , which is approximately 55 nucleotides downstream of the dominant transcription start site (denoted by the highest CAGE peak in FANTOM5 data) in human and mouse (50\u00a0= 1.56\u00a0\u00d7 10\u22129) (50\u00a0= 70.3\u00a0\u00d7 10\u22129) was sensitive to the presence of unlabeled RNA-oligo and ncHOXD11 and coding transcripts are the most negatively correlated, both of which are previously reported target genes along with simultaneous deletion of both CNEs alters HOTAIR-dependent regulation in cis and trans.Our conclusion is based on analyses of large-scale genomics data, thus future work is needed to validate the predicted models All methods can be found in the accompanying"} +{"text": "Salmonella is a gram-negative rod-shape bacterium from the family of Enterobacteriaceae that can cause a wide range of human disease such as enteric fever, gastroenteritis and bacteremia. Here we sequenced two genomes of Salmonella bacteria isolated from the Gallus gallus domesticus host. Genomic DNA of the two Salmonella isolates were extracted and subjected to whole genome sequencing using Illumina platform. The draft genome size of the two Salmonella isolates was determined to be 4,902,295\u202fbp (S18) and 4,847,310\u202fbp (S20) respectively. The percentage of GC content for both draft genomes is the same which is 52.1%. Both the whole genome shotgun project (S18 and S20) has been deposited in National Center for Biotechnology Information Sequence Read Archive under the accession number of SRR7503041 (S18) and SRR7503040 (S20). The sequenced genome (S18 and S20) were aligned with the reference genome and three other Salmonella genomes from serogroup B, D and E. The data obtained show the presence of unique DNA sequences in S18 and S20 genomes. This unique DNA sequences are from the fimbrial gene group. Specifications TableSalmonella genomeThe comparative genomic data show the presence of serogroup-specific DNA fragment in the sequenced (S18 and S20) Salmonella serogroups using molecular methodData obtained can be utilized for identification of Salmonella serogroup C typeThese data can be used for the development of genetic marker in detecting 1Salmonella isolates (S18 and S20) from serogroup C type. Raw reads from both genome were trimmed and assembled. The genome size for S18 is 4902,295\u202fbp while the genome size for S20 is 4847,310\u202fbp. The GC content for both genome are the same which is 52.1%. After genome assembly the S18 samples yielded 225 contigs while the S20 samples yielded 284 contigs. Both assembled genomes were uploaded to the Rapid Annotation using Subsystem Technology (RAST) server for annotation. Both draft genomes from S18 and S20 were then aligned with a reference genome and three other Salmonella genomes from serogroup B, D and E type using CGView Comparison Tool (CCT).The data reported here are the genome sequencing, assembly, annotation and comparative genomic of two The genome annotation using Rapid Annotation using Subsystem Technology (RAST) server classified the S18 draft genome into 403 subsystems with 5005 numbers of coding sequences and 92 numbers of RNAs genes . Meanwhi22.1Salmonella were serogroup following Jiksing et\u00a0al. (2020) (in press) Salmonella isolates (S18 and S20) were cultured overnight on the nutrient agar. These bacterial cultures were then serogrouped using the Salmonella Sero-Quick Group Kit following the manufacturer protocol. This kit comprised several type of antisera which is used to detect the antigen present on Salmonella surface using the slide agglutination method to identify the serogroup type. Two Salmonella isolates (S18 and S20) from the predominant serogroup (serogroup C) were then cultured overnight for genomes extraction. Genome extraction was conducted using Wizard\u00ae Genomic DNA Purification Kit following the manufacturer protocol. Library preparation using the New England Biolabs Next Ultra DNA Library Kit (350\u202fbp insert) was conducted prior to sequencing process. The genomes was then sequenced using HiSeq 4000 sequencer (Illumina platform) for 150 paired-end reads.Prior to DNA extraction, 2.2De novo assembly for both genomes (S18 and S20) were conducted using SPAdes software. The genome assembly quality was assessed using Quality Assessment Tool for Genome Assemblies (QUAST) software Raw reads for both genomes (S18 and S20) were trim with trimmomatic software prior to genome assembly. The authors declare that they have no known competing financial interests or personal relationships which have, or could be perceived to have, influenced the work reported in this article."} +{"text": "This Correction follows an Expression of Concern relating to this article previously published by Portland Press.The authors of the original article above article \u201cmicroRNA-613 exerts anti-angiogenic effect on nasopharyngeal carcinoma cells through inactivating the AKT signaling pathway by downregulating FN1\u201d (Biosci Rep (2019) 39(7), 10.1042/BSR20182196) would like to correct"} +{"text": "Myeloproliferative neoplasms are characterized by mutations in JAK2, MPL and CALR genes. Commonly in diagnostics and previous studies mainly sequencing and common PCR techniques under conventional detection limits are used.Splanchnic vein thromboses are rare, but often appear associated with myeloproliferative neoplasms and represent serious complications.Herein, blood from patients with abdominal vein thromboses in Mecklenburg-West Pomerania , included in an ongoing prospective prevalence study, was analyzed by next generation sequencing representing the complete protein coding regions of JAK2, MPL and CALR genes with a coverage of >\u20092000 reads, therefore an ultradeep targeting approach.JAK2 V617F mutations were detected in 11/44 patients. In four of these cases allele frequencies ranged below the conventional cut off of 2%. MPL W515R was detected in 3/44 cases in low frequencies.Very low allele frequencies of JAK2 and MPL variants in patients with abdominal vein thromboses may indicate early manifestations of myeloproliferative neoplasms.The online version contains supplementary material available at 10.1186/s40364-020-00254-9. To the editor:Splanchnic vein thromboses (SVT) are discussed as first manifestations of myeloproliferative neoplasms (MPN) . JAK2 V6Ultradeep NGS combines the advantages of high sensitivity and of simultaneously analyzing multiple gene loci. We analyzed the prevalence of JAK2, MPL and CALR variants affecting the respective protein coding sequences using an ultradeep targeted sequencing approach.A prospective prevalence study was started in Mecklenburg-West Pomerania to elucidate the prevalence of JAK2 mutations in patients with abdominal vein thromboses. The study included patients from Mecklenburg-Western Pomerania who suffered from current or previous abdominal vein thromboses. The patient\u2019s written consent and a completed clinical questionnaire was required to analyze the blood sample within the study. There were no exclusion criteria. In blood samples from 44 patients with abdominal vein thromboses we detected in thirteen cases JAK2 V617F and MPL W515R variants including seven mutations below the conventional NGS detection limit of 2%: four JAK2 V617F variants as well as three MPL W515R variants (Table 1/13 (TabMost previous studies in patients with SVT reported the prevalence of JAK2 V617F mutation, seldom informing about the sensitivity of the applied method . By ultrThe existence of low and ultralow cancer associated VAF is of interest as the lately identified ARCH may represent a pre-manifestation event in patients showing no further criteria for hematologic neoplasms . Follow-In view of highly sensitive molecular diagnostic tools conventional diagnostic criteria and definition of pathological conditions/diseases may need reconsideration.Additional file 1."} +{"text": "BRCA mutants show better responses to platinum based agents, however, such tumors can utilize the poly(adenosine diphosphate [ADP]\u2013ribose) polymerase (PARP) pathway as a salvage mechanism. Therefore, inhibition of PARP pathway could lead to tumor destruction and synthetic lethality in presence of BRCA mutation. Various PARP inhibitors have been approved for treatment of patients with germline or somatic BRCA mutant breast and ovarian cancer. This provides basis of using PARP inhibitors in patients with pancreatic cancer that harbor BRCA mutation. A recent phase III Pancreas Cancer Olaparib Ongoing (POLO) study showed impressive results with near doubling of progression free survival compared to placebo (7.4 vs 3.8 months). These results highlight the importance of germline testing for all patients with pancreatic cancer and inclusion of additional deficiencies in homologous recombination repair (ATM and PALB2) including BRCA variants of uncertain significance should be further explored.Survival rates for pancreatic cancer remain dismal. Current standard of care treatment regimens provide transient clinical benefit but eventually chemoresistance develops. Tumors deficient in deoxyribonucleic acid (DNA) damage repair mechanisms such as Given the subtle clinical presentation, most patients have advanced disease at the time of diagnosis. Hence, the 5-year-survival rate is 3% and the median overall survival (OS) is around 6-months for patients with metastatic disease.KRAS mutation is crucial for pancreatic carcinogenesis and more than 90% of pancreatic tumors express KRAS mutated protein.7 Inactivation of cyclin-dependent kinase inhibitor 2A (CDKN2A), SMAD4, TP53, and other tumor suppressor genes are also key elements implicated in this progression model.Mechanisms for chemotherapy resistance may be related to tumor microenvironment or intrinsic genetic alterations. A proposed mechanism for development of PDAC includes multiple steps where premalignant lesions called pancreatic intraepithelial neoplasia (PanIN), intraductal papillary mucinous neoplasm (IPMN), and mucinous cystic neoplasm (MCN) develop into carcinoma. 8 Importantly, studies have demonstrated the clinical benefit of PARP inhibition in carriers of BRCA1 or BRCA2 mutation.9Additionally, genes involved in deoxyribonucleic acid (DNA) damage repair (DDR) may also contribute to the pathogenesis of PDAC and deficiency in DDR mechanism leads to an increased risk of cancer. It is known that poly(adenosine diphosphate [ADP]\u2013ribose) polymerase (PARP) 1/2 detect DNA damage and promote its repair.BRCA1, BRCA2, PALB2, or ATM have been associated to have better responses to platinum based chemotherapy agents. This is explained by the fact that platinum compounds generate double-strand DNA breaks that cannot be repaired due to mutation in double-strand DNA repair genes. However, tumors that harbor mutations in homologous recombination genes, can utilize the Poly (ADP-ribose) polymerase (PARP) pathway that is involved in single strand DNA break repair as a salvage mechanism to repair DNA damage. Therefore, inhibition of PARP mediated pathway could lead to tumor destruction and synthetic lethality in presence of BRCA mutations of whom were known carriers of a BRCA 1 or BRCA 2 mutation, received olaparib at escalating doses. This study recruited patients 18-years of age or older with treatment refractory solid tumors. Dosing of olaparib was started at 10 mg daily for two of every three weeks, and was doubled every cycle of treatment, if tolerated. This trial demonstrated that the maximum tolerated dose of olaparib to be 400 mg twice daily. The most common side effects included grade 1\u20132 nausea, and fatigue in one third of patients. The most common grade 3\u20134 toxicities were lymphopenia and anemia, which occurred in up to 5% of the study population. Overall, 23 patients with BRCA mutation were treated and 19 patients were evaluated for response. 12 of 19 (63%) patients had clinical benefit defined as radiologic response based on response evaluation criteria in solid tumors (RECIST), tumor-marker responses defined as a decline in the tumor-marker level of more than 50% that was sustained for at least 4-weeks, or stable disease for a period of 4-months or more.9Phase I studies were conducted after having demonstrated synthetic lethality with PARP inhibitors using BRCA1 or BRCA2 mutations. In a study of 298 patients with pre-treated solid tumors, 23 of whom had pancreatic cancer, patients received olaparib at 400 mg twice daily. The response rate was found to be 26% in all patients and 22% in patients with pancreatic specifically.13 Another multicenter phase II study, RUCAPANC, enrolled patients with BRCA1 or BRCA2 mutations and pancreatic cancer. In this study, 19 patients were enrolled and treated with the PARP inhibitor Rucaparib at 600 mg twice a day. Clinical responses were seen in 3/19 (16%) of the patients.14 Currently there are no head to head trials comparing various PARP inhibitors comparing efficacy. A meta-analysis presented at the Society of Gynecologic Oncology (SGO) meeting 2018 showed no difference in efficacy among the three PARP inhibitors but demonstrated a more favorable safety profile for olaparib, associated with a reduced odds ratio of grade 3 or greater adverse events and treatment interruption.15Following promising phase I trial results, several phase II trials were conducted looking at PARP inhibitors in patients with germline 16 Patients with metastatic pancreatic cancer enrolled for this randomized, double-blind, placebo-controlled phase III trial must have harbored a deleterious germline mutation of BRCA1 or BRCA2 and received at least 16-weeks of continuous first-line platinum-based chemotherapy for eligibility. Patients were randomly assigned in a 3:2 ratio to receive olaparib 300 mg twice daily or placebo (PBO) as a maintenance therapy started within 4\u20138 weeks after the final dose of first-line chemotherapy. Of 3315 patients screened for eligibility, 154 underwent randomization with 92 assigned to receive olaparib and 62 to receive placebo. The majority (>80% in each group) received a variant of FOLFIRINOX with an option to hold the platinum component after 16-weeks if toxicities arised. Although the duration of first-line chemotherapy was not limited, the majority in each group received therapy ranging from 16-weeks to 6-months .The results of a recently published phase III Pancreas Cancer Olaparib Ongoing (POLO) trial were chosen for presentation at the plenary session of the American Society of Clinical Oncology (ASCO) 2019 meeting, highlighting the great excitement for the emerging roles of PARP inhibitor therapy in numerous solid tumors.vs. 3.8 mo, HR 0.53, 95% CI 0.35\u20130.82, p=0.004), generating an enthusiasm shown similar to the practice-changing SOLO1 olaparib maintenance trial for newly diagnosed advanced ovarian cancer (17 Significant responses were seen in 20 patients in the olaparib group (20%) compared to 6 in PBO (10%), with 2 complete responses seen with olaparib alone. Olaparib was very well tolerated with only 5% rate of discontinuation for toxicity with no changes in quality of life compared to PBO as measured by the EORTC QLQ-C30 score. At this interim analysis with data maturity of only 46%, there was no OS benefit yet seen . In addition to the immaturity of the data for this secondary endpoint, PARP inhibitor use in the PBO group after discontinuation of study drug and increased use of chemotherapy upon progression (49% in olaparib group vs. 74% in PBO group) may have contributed to the lack of OS benefit.The study met its primary endpoint impressively with near doubling of progression free survival (PFS) in the olaparib group compared to placebo treated with olaparib 400 mg twice daily identified mutations in DNA repair related genes including BRCA 1/2, ATM, and PALB2 in 16 out of 49 (33%).19 Subgroup analysis per altered gene identified response rates defined as radiologic response per RECIST criteria or greater than 50% reduction in PSA. Response rates of 80%, 57%, and 37% were seen in patients with BRCA 1/2, PALB2, and ATM mutations respectively.19 The highest response rate in PSA reduction was noted in patients with BRCA 1/2 and PALB2 subgroups. This study corroborates the rationale in developing PARP inhibition in DDR-defective patients beyond BRCA mutations and has implications in treatment of PDAC as well.19 Periodic re-evaluation of those found with BRCA variants of uncertain significance should also be warranted for appropriate grouping of this unclear alterations to \u201clikely benign\u201d or \u201clikely pathogenic\u201d classification. The benefits of PARP inhibitor-mediated synthetic lethality for those with germline BRCA1 or BRCA2 mutations are now undeniable and highlights the importance of germline testing for all patients with pancreatic cancer.The growing success in this space calls for further inclusion of those with additional deficiencies of homologous recombination repair, particularly"} +{"text": "Viruses have high mutation rates and generally exist as a mixture of variants in biological samples. Next-generation sequencing (NGS) approaches have surpassed Sanger for generating long viral sequences, yet how variants affect NGS de novo assembly remains largely unexplored.Our results from >\u200915,000 simulated experiments showed that presence of variants can turn an assembly of one genome into tens to thousands of contigs. This \u201cvariant interference\u201d (VI) is highly consistent and reproducible by ten commonly-used de novo assemblers, and occurs over a range of genome length, read length, and GC content. The main driver of VI is pairwise identities between viral variants. These findings were further supported by in silico simulations, where selective removal of minor variant reads from clinical datasets allow the \u201crescue\u201d of full viral genomes from fragmented contigs.These results call for careful interpretation of contigs and contig numbers from de novo assembly in viral deep sequencing. For many years, Sanger sequencing has been used to complement classical epidemiological and laboratory methods for investigating viral infections . As tech\u2212\u20093 mutations per nucleotide, per genomic replication cycle whose aArtificial genomes were constructed for four genome lengths: 2 Kb, 10 Kb, 100 Kb, and 1\u2009Mb, with varying GC content from 20 to 80%, in 5% increments was used to calculate Pearson\u2019s correlation and Spearman\u2019s \u03c1 values to compare the association between percent GC levels and the number of contigs produced at each PID level. Since there was little statistical difference when comparing the contig numbers generated at varying percent GC for each of the four genome length datasets and 494 assemblies for the poliovirus and coronavirus datasets (247 datasets per genome X 2 genomes X 1 assembler) , the finGenome variants were generated as described above (\u201cCreation of simulated variant genomes and reads\u201d) for a genome of size 100 Kb with 50% GC; this was the starting initial variant genome. In this simulation, initial and mutation variant reads at four sequencing read lengths were created using ART. A total of 538 SPAdes assemblies were generated were analyzed for this experiment, as previous sequencing analysis using Geneious indicated the presence of genome variants. The datasets were analyzed using an in-house pipeline (VPipe), which pe50% [De novo assembly for each of the four clinical samples was performed for the following binned NGS datasets: (1) total reads only (T); (2) major variants only (M); (3) major and minor variants only (Mm); and (4) major variants and background reads only (MB). This was repeated with three assembly programs: SPAdes, Cap3, and Geneious. The length of the longest contig produced from each assembly and the performance metric UG50% were calAdditional file 1. Supplemental Text: Analysis of viral GenBank records demonstrated the advent of NGS in viral sequencing in the last two decades. Supplement Figure S1. Workflow diagrams of simulated data from data creation through de novo assembly. Supplement Figure S2. Analysis of the final contig assembly graphs for a clinical sample containing enterovirus A71 (EV-A71) variants using Bandage. Supplement Figure S3. Assembly with three simulated variants. Supplement Table S1. Total counts from NCBI\u2019s GenBank non-redundant nucleotide database. Supplement Table S2. Total count of sequencing technologies for sequences >2000 nt in the NCBI GenBank non-redundant nucleotide database for years 2012\u20132019. Supplement Table S3. Total counts from NCBI\u2019s GenBank non-redundant nucleotide database with multiple sequencing technologies listed per entry. Supplement Table S4. Total counts from NCBI\u2019s GenBank non-redundant nucleotide database of all entries with three and four sequencing technologies listed. Supplement Table S5. Total count of assembly programs used to generate sequences >2000 nt in the NCBI GenBank non-redundant nucleotide database. Supplement Table S6. The 10 de novo assemblers used for analysis of the simulated data, as categorized by their underlying assembly algorithms. Additional file 2. Number of contigs generated by SPades using the simulated data of two variants differed from 75% PID to 99.6% PID, across 20%-80% GC content."} +{"text": "Correction to: Acta Neuropathol Commun (2020) 8:72https://doi.org/10.1186/s40478-020-00947-0In the original publication of this article the refeIn this correction article the reference & citations have been published to rectify this omission.Incorrect:\u201cWe observed increased expression of CSF-2 in gray matter compared to affected white matter, which may contribute to selective vulnerability of white matter in HDLS\u201dCorrect:\u201cPreviously we observed increased expression of CSF-2 in the gray matter but not the white matter of HDLS patientswhich may explain the selective vulnerability of white matter microglia in HDLS\u201d.AbstractDiscussionPage 11Incorrect:\u201cThe unique vulnerability of frontal white matter in HDLS may also be accounted for by differences in abundance of the two main microglial trophic factors between brain regions, with CSF-2 being significantly up-regulated in gray matter compared to white matter and vice versa for CSF-1 [26].\u201dCorrect:\u201cThe unique vulnerability of frontal white matter in HDLS may also be accounted for by differences in abundance of microglial trophic factors between brain regions. Compared to healthy individuals, ALSP patients significantly up-regulate the expression of with CSF-2 in gray matter but not in the white matterand this may compensate for reduced signaling via the CSF-1R in microglia.\u201dPage 12Incorrect:\u201cCombined with developmental issues highlighted in Csf1r+/\u2212 mice, where it has been suggested that aberrant energy metabolism may play a pathogenetic role, additional studies are warranted on these factors in HDLS [9].\u201dCorrect:\u201cCombined with developmental issues highlighted in Csf1r+/\u2212 mice, where it has been suggested that aberrant energy metabolism may play a pathogenetic role, additional studies are warranted on these factors in HDLS.\u201d"} +{"text": "Desulfovibrio vulgaris strain Hildenborough, and marine bacterium Desulfovibrio alaskensis strain G-20) grown on lactate across a range of metabolic rates and ambient sulfate concentrations. We use a combination of experimental sulfate oxygen isotope data and nonlinear regression fitting tools to solve for unknown kinetic, step-specific oxygen isotope effects. This approach enables identification of key isotopic reactions within the metabolic pathway, and defines a new, calibrated framework for understanding oxygen isotope variability in sulfate. This approach is then combined with porewater sulfate/sulfide concentration data and diagenetic modeling to reproduce measured 18O/16O in porewater sulfate. From here, we infer cell-specific sulfate reduction rates and predict abundance of active cells of sulfate reducing bacteria, the result of which is consistent with direct biological measurements.The majority of anaerobic biogeochemical cycling occurs within marine sediments. To understand these processes, quantifying the distribution of active cells and gross metabolic activity is essential. We present an isotope model rooted in thermodynamics to draw quantitative links between cell-specific sulfate reduction rates and active sedimentary cell abundances. This model is calibrated using data from a series of continuous culture experiments with two strains of sulfate reducing bacteria (freshwater bacterium A significant fraction of biogeochemical cycling takes place within marine sediments and is driven by microbial activity , 2. MicrIn certain cases, cell-specific metabolic rates can be inferred via the inorganic chemical signatures of that activity. One such example, and central to this study, are the laboratory calibrated stable isotope effects associated with microbial sulfate reduction (MSR) , 12. ThiWe aim to quantitatively link the MSR biogeochemistry and its oxygen isotope effect. Mathematical frameworks have been developed to establish quantitative links between cell-specific parameters chemical environment in static) experiments covering a range of growth rates and ambient sulfate concentrations using two strains of sulfate reducing Bacteria . We use these data to calibrate an isotope model for MSR 18O/16O fractionation rooted in reaction thermodynamics. This model is adapted from a similar model focused on describing MSR sulfur isotope fractionation . En. En48]. ace Fig.\u00a0. From heace Fig.\u00a0. This anace Fig.\u00a0.Fig. 6Me2S per cell per day) and incorporating the organoclastic sulfate reduction rate (\u03bcmol S per cm3 per year) from diagenetic model studies at the same site and depth [3). Aarhus Bay was specifically chosen for this comparison given that active sedimentary cell abundances were independently calculated using molecular tools [Calculating active cell abundances then simply requires csSRR to be merged with diagenetic model rate estimates. This is done via a straightforward unit conversion . Our model effectively reproduces observed sulfate fate Fig.. Here, tSupplementary MaterialSupplementary Figure 1Supplementary Figure 2Supplementary Figure 3Supplementary Figure 4Supplementary Figure 5Supplementary Figure 6Supplementary Figure 7Supplementary Figure 8Supplementary Figure 9Supplementary Figure 10Supplementary Table 1Supplementary Table 2"} +{"text": "Digalu, there is a marked increase in disease prevalence following the incursion of new rust races into Ethiopia, which indicates a pronounced boom-and-bust cycle of major gene resistance. Using spatial analyses, we identify hotspots of disease risk for all three rusts, show a linear correlation between altitude and disease prevalence, and find a pronounced north-south trend in stem rust prevalence. Temporal analyses show a sigmoidal increase in disease levels during the wheat season and strong inter-annual variations. While a simple logistic curve performs satisfactorily in predicting stem rust in some years, it cannot account for the complex outbreak patterns in other years and fails to predict the occurrence of stripe and leaf rust. The empirical insights into wheat rust epidemiology in Ethiopia presented here provide a basis for improving future surveillance and to inform the development of mechanistic models to predict disease spread.Wheat rusts are the key biological constraint to wheat production in Ethiopia\u2014one of Africa\u2019s largest wheat producing countries. The fungal diseases cause economic losses and threaten livelihoods of smallholder farmers. While it is known that wheat rust epidemics have occurred in Ethiopia, to date no systematic long-term analysis of past outbreaks has been available. We present results from one of the most comprehensive surveillance campaigns of wheat rusts in Africa. More than 13,000 fields have been surveyed during the last 13 years. Using a combination of spatial data-analysis and visualization, statistical tools, and empirical modelling, we identify trends in the distribution of wheat stem rust (Sr), stripe rust (Yr) and leaf rust (Lr). Results show very high infection levels . These recurrent rust outbreaks lead to substantial economic losses, which we estimate to be of the order of 10s of millions of US-D annually. On the widely adopted wheat variety, In some areas, wheat is also grown in the preceding short rainy (belg) season, from March or April to June or July. In the main wheat producing areas of Ethiopia , the environment is suitable for infection with wheat rusts almost year around ; (iii) moderate incidence (severity) [20\u201340%]; (iv) high incidence (severity) [>40%]. In the analyses, we considered both disease measures, incidence and severity, for all three types of wheat rusts with the aim of capturing key aspects of wheat rust epidemiology in Ethiopia.The in 2007 . The sur in 2007 . Two meabb scale taking ahttps://opendatakit.org/) for digital collection of surveillance results in near-real time. Selected rust samples collected during the surveillance campaigns were analysed by dedicated laboratories to determine races present using standard methodology and nomenclature , is obtained as a function of the annual sample mean disease incidence (considering only moderate and high infections), i, and the annual FAO statistic for total national wheat areas in Ethiopia in Ethiopia, as reported in FAO statistics, and the fraction of yield lost to a specific rust type, r. The fraction of yield lost to rust, r, is obtained as a function of field-scale disease severity, s, and wheat growth stage, g, combined with published empirical results ).(DOCX)Click here for additional data file.S6 Fig(A) wheat stripe rust; (B) wheat stem rust; (C) wheat leaf rust. A set of longitude-intervals are defined covering all longitudes at which wheat is grown in Ethiopia (x-axis). The surveys from all years are grouped according to their longitude coordinate and the aggregated mean prevalence is calculated for each longitude interval. (top row) long-term mean prevalence of incidence scores per longitude, (bottom row) long-term mean prevalence of severity scores per longitude. The labels at the top of the x-axes show the total number of surveys per latitude bin. Prevalence is calculated as: [number of surveys with incidence score x / total number of surveys per altitude-interval]).(DOCX)Click here for additional data file.S7 Fig(A) wheat stripe rust; (B) wheat stem rust; (C) wheat leaf rust. The labels at the top of the x-axes show the total number of surveys per year. Prevalence is calculated as: [number of surveys with disease incidence (severity) / total number of surveys per wheat variety].(DOCX)Click here for additional data file.S8 Fig(A) wheat stripe rust; (B) wheat stem rust; (C) wheat leaf rust. Symbols show sample mean prevalence levels in surveys and lines are linear fits to the mean prevalence levels (obtained using the MATLAB function fitlm). For each wheat rust and each disease metric it was tested if the slope of the fitted linear model is different from a constant line with a slope coefficient of zero using the MATLAB function CoefTest. According to this test there is no statistically significant decrease or increase in the mean wheat stripe rust and wheat stem rust disease prevalence during years 2010\u20132019. The resulting p-values for the F-Test that the slope coefficient of the linear model is different to zero are larger than a significance level of 0.05 for all linear models of stripe rust and stem rust. However, there is a statistically significant decrease of wheat leaf rust prevalence over the years. The p-values for the F-Test of the slope of all leaf rust models are all smaller than 0.05 except for the linear fit to high severity levels.(DOCX)Click here for additional data file.S9 Fig(A) wheat stripe rust; (B) wheat stem rust; (C) wheat leaf rust. The maps show disease incidence at all survey points of three exemplar years: 2010, 2014 and 2019. Symbols: green\u2014no disease; yellow\u2014low incidence; orange\u2014moderate incidence; red\u2014high incidence; grey areas\u2014wheat producing regions. In 2010 a major wheat stripe rust epidemic led to infections covering large parts of western and central Ethiopian wheat producing areas . In these regions, wheat stem rust incidence was low in 2010 . In 2014 a major wheat stem rust epidemic occurred in large parts of southern and central Ethiopia . In these areas wheat stripe rust incidence was low . See (DOCX)Click here for additional data file.S10 Fig(A) wheat stripe rust; (B) wheat stem rust; (C) wheat leaf rust. The maps show disease severity at all survey points of three exemplar years: 2010, 2014 and 2019. Symbols: green\u2014no disease; yellow\u2014low incidence; orange\u2014moderate incidence; red\u2014high incidence; grey areas\u2014wheat producing regions. Maps created using R as GIS [R as GIS \u201322.(DOCX)Click here for additional data file.S11 Fig(A) wheat stripe rust incidence at the beginning (left map), middle (centre map) and end (right map) of the main wheat season 2010; (B) wheat stem rust incidence at the beginning (left map), middle (centre map) and end (right map) of the main wheat season 2014; (C) wheat leaf rust incidence at the beginning (left map), middle (centre map) and end (right map) of the main wheat season 2010. See The maps show disease incidence at all survey points at three different times of the main wheat season. Symbols: green\u2014no disease; yellow\u2014low incidence; orange\u2014moderate incidence; red\u2014high incidence; grey areas\u2014wheat producing regions. (DOCX)Click here for additional data file.S12 Fig(A) wheat stripe rust severity at the beginning (left map), middle (centre map) and end (right map) of the main wheat season 2010; (B) wheat stem rust severity at the beginning (left map), middle (centre map) and end (right map) of the main wheat season 2014; (C) wheat leaf rust severity at the beginning (left map), middle (centre map) and end (right map) of the main wheat season 2010. Maps created using R as GIS [The maps show disease severity at all survey points at three different times of the main wheat season. Symbols: green\u2014no disease; yellow\u2014low incidence; orange\u2014moderate incidence; red\u2014high incidence; grey areas\u2014wheat producing regions. R as GIS \u201322.(DOCX)Click here for additional data file.S13 Fig(A) wheat stripe rust; (B) wheat stem rust; (C) wheat leaf rust. Two simple logistic models were used to predict moderate or high wheat rust incidence cases: A temporal model Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S1 Movie(MP4)Click here for additional data file.S2 Movie(MP4)Click here for additional data file.S3 Movie(MP4)Click here for additional data file.S4 Movie(MP4)Click here for additional data file.S5 Movie(MP4)Click here for additional data file.S6 Movie(MP4)Click here for additional data file.S7 Movie(MP4)Click here for additional data file.S8 Movie(MP4)Click here for additional data file.S9 Movie(MP4)Click here for additional data file.S10 Movie(MP4)Click here for additional data file.S11 Movie(MP4)Click here for additional data file.S12 Movie(MP4)Click here for additional data file.S1 Appendix(DOCX)Click here for additional data file.S2 Appendix(DOCX)Click here for additional data file."} +{"text": "The second sentence of the fifth paragraph of the Results should read as follows: \u201cCompared with 569 individuals in the existing randomized clinical trial,15 940 individuals included in the present analysis were older , more likely to be male (547 [58%] vs 298 [52%]), and less likely to have undergone prior chemotherapy (53 [6%] vs 45 [8%]).\u201d This article has been corrected.1In the Original Investigation titled \u201cEstimates of Overall Survival in Patients With Cancer Receiving Different Treatment Regimens: Emulating Hypothetical Target Trials in the Surveillance, Epidemiology, and End Results (SEER)\u2013Medicare Linked Database,\u201d"} +{"text": "Gastric cancer (GC) is one of the most common cancers in the world with a high ratio of mortality. Regarding the late diagnosis, there is a high ratio of distant metastasis among GC cases. Despite the recent progresses in therapeutic modalities, there is not still an efficient therapeutic method to increase survival rate of metastatic GC cases.Apart from the various intracellular signaling pathways which are involved in tumor cell migration and metastasis, the local microenvironment is also a critical regulator of tumor cell migration. Indeed, the intracellular signaling pathways also exert their final metastatic roles through regulation of extra cellular matrix (ECM). Therefore, it is required to assess the role of extra cellular components in biology of GC.In the present review, we summarize 48 of the significant ECM components including 17 ECM modifying enzymes, seven extracellular angiogenic factors, 13 cell adhesion and cytoskeletal organizers, seven matricellular proteins and growth factors, and four proteoglycans and extra cellular glycoproteins. This review paves the way of determination of a specific extra cellular diagnostic and prognostic panel marker for the GC patients. The local microenvironment has critical role in regulation of cell functions . The extCarcinogens exert their impact through DNA damage, aberrant cellular processes, and microenvironment alteration . The effhttps://www.ncbi.nlm.nih.gov) are also presented in Table\u00a0Gastric Cancer (GC) is the fourth common and the second leading cause of cancer-related deaths in the world . DistantIn this section, we summarize 17 gene products including seven matrix metallopeptidases , laminin subunit gamma 2 (LAMC2), three ADAMTS metallopeptidases , 2 TIMP metallopeptidases , three lysyl oxidase , and tissue factor pathway inhibitor 2 (TFPI2).Matrix metalloproteinases (MMPs), heparanase, cathepsins, urokinase plasminogen activator, and lysyl oxidase are ECM modifying enzymes contributing with tumor metastasis \u201379. DegrLaminin subunit gamma 2 (LAMC2) is expressed in tumor cells and can be used as a marker of malignant epithelial cells . The LAMThe human ADAMTS family of 19 secreted, multidomain proteolytic enzymes is involved in a wide range of biological processes including ECM assembly and degradation, hemostasis, organogenesis and the regulation of angiogenesis . ImmunohTissue inhibitors of metalloproteinases (TIMPs) as the main MMPs endogenous inhibitors bind to the enzyme catalytic site . RegulatTumor progression requires networks of both the immediate (cell\u2013cell or cell\u2013matrix interactions) and extended microenvironment (vascularization). The stromal activation is parallel with tumor cell growth and metastasis , 96. LysTissue factor pathway inhibitor 2 (TFPI2) is a serine proteinase inhibitor involved in protection of ECM from degradation . TherefoIn this section, we summarize seven gene products including extracellular inducer (BSG: EMMPRIN), extracellular angiostatic protein (MMRN2), extracellular matrix glycoprotein (EMILIN2), heparinase (HPSE), collagen triple helix repeat containing 1 (CTHRC1), galectin 1 (LGLS1), and extracellular matrix protein 1 (ECM1).Angiogenesis is a critical process during tumor growth which initiates from the outgrowth and migration of endothelial cells from parental vessels through vascular endothelial growth factor (VEGF). Cytokines, growth factors, and ECM proteins are important regulators of angiogenesis , 106. InMultimerin 2 (MMRN2) is an extracellular angiostatic protein which suppresses the VEGFA/VEGFR2 signaling pathway . It has Collagen triple helix repeat containing 1 (CTHRC1) is a glycoprotein containing a short collagen triple helix repeat associated with cell migration through regulation of collagen matrix deposition . It alsoExtracellular matrix protein 1 (ECM1) is involved in cell proliferation, migration, angiogenesis, and EMT . During In this section, we summarize 13 gene products including a member of the protocadherin alpha gene cluster (PCDH8), laminin subunit gamma 2 (LAMC2), a claudin family member (CLDN4), cellular adhesion proteins (LLGL1 and ANOS1), fibronectin family members (FN1 and FNDC1), integrin alpha chain member (ITGA4), immunoglobulin family [ICAM1 (CD54)], cytoskeletal reorganizer (CFL1), hyaluronan mediated motility receptor (HMMR), microfibril associated protein (MFAP2), and an integrin ligand (SPON2).Epithelial cell junctions have various functions such as communication, anchoring, and sealing. E-cadherin is the main protein of adherent junctions to stabilize the basolateral cell-cell contact. Therefore, E-cadherin loss increases cell migration . ProtocaLLGL1 is also a cellular adhesion protein that is involved in epithelial cell polarity and migration through a polarity complex comprising of PAR6, PAR3, PKC, and LLGL , 144. ItThe Anosmin 1 (ANOS1) is a cell adhesion protein associated with migration of gonadotropin-releasing hormone neurons. It increases the malignant behavior through integrin signaling pathways in brain tumors and promotes cell migration and drug resistance in colon cancer , 154. ItLeptin is an adipokine that is involved in body weight homeostasis through regulation of energy metabolism . Leptin Cofilin 1 (CFL1) is a cytoskeletal reorganizer and EMT regulator in tumor cells . It has Microfibril associated protein 2 (MFAP2) is one of the main regulators of microfibril function which binds with ECM and TGF-\u03b2 through specific domains in carboxyl and amino-terminal, respectively , 168. ItIn this section, we summarize seven matricellular protein gene products including secreted protein acidic and cysteine rich (SPARC), SPARC like protein 1 (SPARCL1), serpin family E member 1 (SERPINE1), fibroblast growth factor receptor 2 (FGFR2), thrombospondin 1 (THBS1), latent transforming growth factor beta binding protein 2 (LTBP2), and periostin (POSTN).Matricellular proteins are ECM glycoproteins that includes osteopontin, tenascins, POSTN, SPARC, and thrombospondins that are associated with embryogenesis, stem cell niches, and tissue remodeling , 170. MoLTBP2 is belonged to the ECM proteins which has calcium-binding epidermal growth factor like and TGF-\u03b2 binding domains . LTBPs hIn this section, we summarize four matricellular protein gene products including versican (VCAN), asporin (ASPN), lumican (LUM), and mucin 2 (MUC2).The ECM is composed of various proteins such as proteoglycans (PGs) and collagens that organize physiological and pathological functions . ProteogAsporin (ASPN) is one of the members of small Leucine-rich proteoglycan (SLRP) family that is associated with ECM modification . It has Mucins are glycoproteins comprising of a mucin core and O-linked oligosaccharides produced by epithelial tissues which are involved in epithelial surfaces protection, inflammation, renewal, and tumorigenesis . AlthougRegarding the importance of extra cellular matrices and components in GC metastasis and invasion, in present review we summarized 48 of significant ECM related factors which have been reported among GC patients. It has been shown that the intra cellular signaling pathways associated with tumor invasion also exert their role through regulation of local ECM. Therefore, according to the lower complexity of ECM compared with intracellular signaling pathways, the ECM can be a more reliable tumor therapeutic target with lower side effects. Indeed this review paves the way of introducing an extra cellular based prognostic and diagnostic panel marker for the GC patients."} +{"text": "Bioscience Reports (2018) 38(5); https://doi.org/10.1042/BSR20180562) would like to provide a correction to The authors of the original article \u201cHigh LINC01605 expression predicts poor prognosis and promotes tumor progression via upregulation of MMP9 in bladder cancer\u201d (The authors apologise for any inconvenience that this error has caused to the readers of their original paper."} +{"text": "This cohort study examines the association of thromboelastographic results with hypercoagulability among patients with coronavirus disease 2019 who have been admitted to an intensive care unit. Disseminated intravascular coagulopathy and other COVID-19\u2013associated coagulopathies occur among patients with severe SARS-CoV-2 infections.STROBE) reporting guidelines for a cohort study.This cohort study was approved by the Baylor College of Medicine institutional review board with a waiver of informed consent granted because this was a retrospective electronic health record review of data collected for clinical purposes. The cohort included all patients admitted to the ICU of Baylor St. Luke's Medical Center from March 15 to April 9, 2020, with SARS-CoV-2 infection confirmed by reverse transcription\u2013polymerase chain reaction test of nasopharyngeal swab. This study is reported following Strengthening the Reporting of Observational Studies in Epidemiology (All patients received standard deep vein thrombosis chemoprophylaxis on ICU admission and therapeutic anticoagulation (heparin infusion or enoxaparin [2 mg/kg/d]) for thrombotic complications. All patients underwent TEG and TEG with heparinase correction on ICU admission. Hypercoagulability was defined as elevated fibrinogen activity greater than a 73\u00b0 angle or maximum amplitude (MA) more than 65 mm on TEG with heparinase correction.P values were 2-sided, and statistical significance was set at .05. Data were analyzed from March 21 to April 14, 2020.Group differences were analyzed using Fisher exact test. Analyses were conducted using SAS statistical software version 9.4 (SAS Institute). This cohort study included 21 patients . Among these patients, 20 (95%) had comorbidities, with a mean (SD) of 3 (2) comorbidities each . Mean (SD) follow-up was 11 (4) days. Regrading thromboembolism risk, 4 patients (19%) had atrial fibrillation, a history of malignant tumors, or chronic kidney disease. Four patients (19%) required extracorporeal membrane oxygenation, and 18 patients (86%) required renal replacement therapy. There were 2 mortalities (10%), both occurring as pulseless electrical activity after acute-onset pulmonary hypertension.Cohort mean international normalized ratio (INR), partial thromboplastin, and platelet levels were within reference ranges, but fibrinogen and dimerized plasmin fragment D levels were elevated . A totalP\u2009=\u2009.01). Elevated MA was observed in 10 patients (100%) in the high event rate group vs 5 patients (45%) in the low event rate group. Innate TEG MA provided 100% sensitivity and 100% negative predictive value (There were no statistically significant differences in prothrombin time, INR, partial thromboplastin time, or platelet levels between 10 patients with at least 2 thrombotic events vs 11 patients with fewer than 2 events . In compve value .5 Underdiagnosis or undertreatment of hypercoagulation may explain the high incidence of unexplained COVID-19 mortalities. These may be associated with potentially preventable microvascular and macrovascular thromboses and consequent cardiovascular complications, including myocardial injury and infarction.6 Accordingly, our institution and other health care systems have adopted immediate full heparinization in patients with high-acuity COVID-19.This cohort study found that higher thromboses rates were associated with TEG results outside reference ranges among patients with COVID-19 who were critically ill. Risk associated with TEG results outside reference ranges manifested as a 62% thrombosis event rate, 2-fold the thrombosis event rates that have been previously reported, despite our use of recommended deep vein thrombosis prophylaxis.6 Similarly, angiotensin II\u2013mediated pulmonary vasoconstriction can lead to stasis and hypercoagulability, as can COVID-19 induction of antiphospholipid antibodies and complement during cytokine storms, causing vasculitis and microthromboses.Hypercoagulation associated with COVID-19 may be due to increased angiotensin II expression secondary to angiotensin-converting enzyme 2 receptor binding and consequently increased plasminogen activator inhibitor C-1 expression, which is consistent with our observation of reduced fibrinolysis in our high thrombotic event rate group.Our finding of INR, partial thromboplastin time, and platelet levels within or close to reference ranges but elevated fibrinogen and dimerized plasmin fragment D levels reflect a complex inflammatory and hematologic profile distinct from the disseminated intravascular coagulopathy associated with COVID-19. In this context, TEG may be critical in accurately identifying patients at increased thrombosis risk and thereby avoiding unnecessary anticoagulation in patients with low thrombosis risk. Specifically, a hypercoagulable innate TEG MA yielded 100% sensitivity and 100% negative predictive value for the occurrence of multiple thromboses.6 Our findings suggest that alterations of diagnostic and prophylactic treatment guidelines may be critical for the successful treatment of coagulopathies associated with COVID-19.One study limitation is whether this retrospective study reflects differences in our anticoagulation practices vs other institutions or their underreporting of thrombotic events, as recently suggested."} +{"text": "The ST-4821 complex (cc4821) is a leading cause of serogroup C and serogroup B invasive meningococcal disease in China where diverse strains in two phylogenetic groups (groups 1 and 2) have acquired fluoroquinolone resistance. cc4821 was recently prevalent among carriage isolates in men who have sex with men in New York City (USA). Genome-level population studies have thus far been limited to Chinese isolates. The aim of the present study was to build upon these with an extended panel of international cc4821 isolates.Neisseria database. Core genome comparisons were performed in PubMLST.Genomes of isolates from Asia (1972 to 2017), Europe (2011 to 2018), North America (2007), and South America (2014) were sequenced or obtained from the PubMLST Four lineages were identified. Western isolates formed a distinct, mainly serogroup B sublineage with alleles associated with fluoroquinolone susceptibility (MIC <0.03 mg/L) and reduced penicillin susceptibility (MIC 0.094 to 1 mg/L). A third of these were from anogenital sites in men who have sex with men and had unique denitrification gene alleles. Generally 4CMenB vaccine strain coverage was reliant on strain-specific NHBA peptides.The previously identified cc4821 group 2 was resolved into three separate lineages. Clustering of western isolates was surprising given the overall diversity of cc4821. Possible association of this cluster with the anogenital niche is worthy of monitoring given concerns surrounding antibiotic resistance and potential subcapsular vaccine escape. Neisseria meningitidis, the meningococcus, is a leading cause of meningitis and sepsis. Six serogroups cause the majority of invasive meningococcal disease (IMD) globally. Prior to 2003/4, meningococcal serogroup A (MenA) was the most common cause of IMD in China with periodic epidemics and . (2018) these we. (2018) The geo-temporal composition and disease status of the four lineages was as follows:The majority of lineage 1 isolates were from China including 16 carrier isolates belonging to MenC , MenB , and serogroup unknown ; 34 invasive isolates belonging to MenB and MenC ; and a single MenB isolate with disease status unknown (2005). The remaining isolate was an invasive MenC isolate from Japan (2017).. Four of the invasive isolates belonged to a distinct cluster of relatively closely related isolates.All lineage 2a isolates were from China and most (n = 45/49) were MenB. The lineage included 44 carrier isolates , n = 1; and unknown, n = 1), and five invasive isolates All of the lineage 2b isolates were from China. The majority (n = 13/15) were carrier isolates including a single MenB (1980), two MenC (1972 and 2007), and ten MenW isolates (2009 to 2017). The two invasive isolates (2002 and 2005) belonged to MenC and formed a distinct sublineage., and a single MenB isolate (2005) with unknown disease status.The majority (n = 40/57) of the lineage 2c isolates were from China including 29 carrier isolates belonging to MenB and MenC , ten invasive isolates belonging to MenB and MenC . These included carrier MenB isolates from meningococcal carriage studies in the USA [. The remaining six lineage 2c isolates were all from anogenital sites among men who have sex with men (MSM). These included three UK MenB isolates that were incidentally isolated from rectal swabs during routine gonococcal screening [Chlamydia trachomatis coinfected sex worker with symptoms of urethritis in Spain . The final isolate belonged to MenC and was a urethral swab isolate from a UK bisexual male practicing protected insertive sex and displaying symptoms of urethritis . Coinfections were ruled out for C. trachomatis, Mycoplasma genitalium and gonococcus.The remaining lineage 2c isolates (n = 17/57) were from nine countries in Europe and North and South America and formed a distinct cluster within lineage 2c designated the \u2018cc4821 lineage 2c RoW (rest of world) cluster\u2019 hildren) ; Brazil hildren) ; Italy were all from China and comprised ten carrier isolates including MenB and MenC isolates; three invasive MenB isolates (1985 to 2011); and two MenB isolates with unspecified disease status (1980 to 1985).gyrA and parC alleles and penA alleles associated with reduced penicillin susceptibility (0.094 to 1 mg/L) . Detailed lineage breakdowns of genotypic antibiotic resistance determinants are depicted in S5 Fig and as follows:The different lineages/sublineages contained varying distributions of isolates with fluoroquinolone resistance-associated (MICs \u2265 0.03 mg/L) gyrA alleles . Among these, 45 isolates (including 18/20 epidemic clone isolates) possessed wild type penA alleles. One of these, a MenB carrier isolate (2009), also possessed a mutant parC allele (mutation S88P). Three isolates, including an invasive MenB (2014) and MenC (2012) isolate and a carrier MenB isolate (2016), possessed mutant penA alleles . A further three carrier isolates (including 2/20 epidemic clone) and one invasive isolate did not have full length penA sequence data. The single Japanese epidemic clone isolate possessed mutant gyrA and parC (mutation S87I) alleles and WT penA alleles.All 52 of the lineage 1 isolates possessed mutant (resistance-associated) gyrA and penA alleles. Of these, two MenB isolates (2014 to 2016) also possessed mutant parC alleles (mutation E91K). A further 22/40 MenB isolates (2006 to 2016), and a single cnl (2009) and serogroup-unknown (2016) isolates possessed mutant gyrA and WT penA alleles. Of these, a single MenB isolate (2013) also possessed a mutant parC allele (mutation S87I). The remaining 10/40 MenB (2005 to 2014) and 1/1 MenC (2005) carrier isolates possessed WT gyrA and penA alleles.Among the lineage 2a carrier isolates, 8/40 MenB isolates (2011 to 2016) and 1/1 MenX isolates (2009) possessed mutant gyrA and penA alleles, two (2011 to 2013) possessed mutant gyrA alleles and WT penA alleles, and two (both 2014) possessed WT gyrA and mutant penA alleles.Among the five MenB lineage 2a invasive isolates, one (2017) possessed mutant gyrA and penA alleles. The other MenC isolate (2007) and 8/10 MenW isolates (2009 to 2017) possessed mutant gyrA and WT penA alleles. The remaining 2/10 (2014 and 2016) MenW isolates possessed mutant gyrA and penA alleles. The two invasive MenC isolates (2002 and 2005) possessed WT gyrA and penA alleles.Among the lineage 2b carrier isolates, 1/1 MenB (1980) and 1/2 MenC (1972) possessed WT gyrA and penA alleles, 12/26 (2005 to 2016) possessed mutant gyrA and WT penA alleles, including one isolate with a mutant (S87I) parC allele, 4/26 (2012 to 2014) possessed WT gyrA and mutant penA alleles, and 1/26 (2013) possessed mutant alleles for both gyrA and penA. All three Chinese MenC carrier isolates (2016 to 2017) possessed mutant alleles for both gyrA and penA. Among the Chinese invasive MenB lineage 2c isolates, 4/9 possessed WT gyrA and penA alleles, 1/9 possessed WT gyrA and mutant penA alleles, 2/9 possessed mutant gyrA and WT penA alleles, and 2/9 possessed mutant alleles for both gyrA and penA. The single Chinese invasive MenC isolate possessed mutant gyrA and WT penA alleles. A further MenB isolate with unknown disease status possessed WT gyrA and penA alleles.Among the Chinese lineage 2c MenB carrier isolates, 9/26 isolates (1977 to 2011) possessed WT gyrA alleles and mutant penA alleles.All 17 lineage 2c RoW isolates possessed WT gyrA and penA alleles. A further 4/8 MenC isolates (2005 to 2008) possessed mutant gyrA and WT penA alleles, while 1/8 MenC isolates (1980) possessed WT gyrA and incomplete penA alleles.Among the diffuse carrier isolates 2/2 MenB (2009 and 2011) and 3/8 MenC isolates (1973 to 1978) possessed WT gyrA and penA. The remaining isolate (2009) possessed mutant gyrA and penA.Among the diffuse invasive isolates, 2/3 MenB isolates (1985 and 2011) possessed WT alleles for both gyrA and penA alleles.The two diffuse isolates with unknown disease status (1980 and 1985) possessed WT nhba sequences and one, a UK invasive rectal swab isolate from 2017, had a frameshifted nhba allele. The remaining 185 isolates possessed alleles for one of 32 intact NHBA peptides, 22 of which were unique to China, and 27 of which were only, or predominantly, observed in cc4821 on the PubMLST Neisseria database (accessed 24/06/20). 4CMenB strain coverage of all but one of the encoded NHBA peptides was unpredictable according to gMATS. The remaining peptide, peptide 10 was predicted by gMATS to be covered and occurred in a single lineage 2a MenB carrier isolate from 2013 . The majority of isolates 144/188 possessed fHbp variant 2 or 3 alleles and a further two isolates, a Chinese invasive MenC lineage 1 (non-epidemic clone) isolate from 2011 and a Chinese lineage 2c MenC carrier isolate from 2017, possessed a truncated allele (nonsense mutation) and a complete deletion of fhbp, respectively. The remaining 42 isolates possessed alleles for one of 13 fHbp variant 1 peptides, eight of which were only, or predominantly, found in cc4821, and six of which were unique to China on the PubMLST Neisseria database (accessed 24/06/20). All 13 peptides afforded unpredictable 4CMenB strain coverage according to gMATS. Given the general absence of gMATS-predictable strain coverage, potential strain coverage was estimated based on the most conservative criteria that any NHBA peptide and any fHbp variant 1 peptide is potentially covered. Potential antigen-specific strain coverage of cc4821 lineages/sublineages by 4CMenB is shown in Fig 4. A detailed breakdown of lineage-specific potential 4CMenb strain coverage is as follows:None of the cc4821/ST-14840 isolates possessed alleles for PorA P1.4 or NadA. Two isolates had incomplete nhba sequences. Interestingly, only 3/16 carrier isolates, including 1/5 epidemic clone carrier isolates (1/4 MenB and 2/10 MenC) were also potentially covered by fHbp. By contrast, 28/35 of the invasive isolates (3/6 MenB and 25/29 MenC), including 13/15 invasive epidemic clone isolates, were potentially covered by fHbp. One invasive MenC (2011) isolate possessed a truncated fhbp allele with a nonsense mutation.At least 50 of the 52 lineage 1 isolates were potentially covered by NHBA though this was heavily reliant on peptide 503 (n = 43) being covered. The two remaining isolates had incomplete All 49 lineage 2a isolates were potentially covered by NHBA though this was heavily reliant on peptides 910 and 688 being covered since these accounted for a half and a quarter of the isolates, respectively. Among the invasive isolates, 2/5 were also potentially covered by virtue of one of two different fHbp peptides.All of the lineage 2b isolates were potentially covered by NHBA, though this was reliant on peptide 697 being covered. Additionally, 1/13 carrier isolates and the two invasive MenC isolates were potentially covered by virtue of fHbp.fhbp gene was lacking in one MenC carrier isolate (2017) and was replaced by N. lactamica sequence as previously reported among proposed ST-286 complex isolates [All of the 40 Chinese lineage 2c isolates were potentially covered by NHBA though this was mainly reliant on coverage of peptides 669 (n = 22) and 697 (n = 9). In addition, 4/25 carrier isolates and 2/10 invasive isolates were potentially covered by fHbp. An isolates .nhba allele. None of the RoW isolates were covered by fHbp.Among the lineage 2c RoW isolates, 16/17 were potentially covered by virtue of NHBA, in particular peptide 669 (n = 15). One MenB isolate from a rectal swab from an invasive disease patient possessed a frameshifted Potential 4CMenB coverage of the 15 diffuse isolates was limited to NHBA, in particular peptide 669 (n = 10).aniA) gene has previously been implicated in the adaptation of meningococci to the anogenital niche [aniA alleles within the cc4821 population structure is shown in S6 Fig. A number of RoW cluster and lineage 2c isolates possessed aniA (NEIS1549) allele 8 that was unique to these and a small number of other ST-3200 (cc4821) related isolates on the PubMLST Neisseria database (accessed 21/07/20). A genome comparison of all lineage 2c isolates in terms of all NEIS loci showed that isolates with aniA allele 8 also possessed a unique nitric oxide reductase allele\u2013allele 379. Most other lineage 2c isolates possessed aniA allele 287 and norB allele 838 in common with diverse isolates from lineages 2a and 2b. The most commonly observed alleles of the loci immediately upstream and downstream of aniA and norB did not differ between the two sets of isolates suggesting a possible recombination event. A nucleotide alignment of aniA, norB and flanking genes between isolates representative of the putative ancestral and putative recombinant states identified a potentially horizontally acquired region of 2022 bp covering 82% of the 5\u2019 end of aniA, 30% of the 5\u2019end of norB, and the 381 bp intergenic region . The extent of the change in the respective peptides was relatively modest comprising P23Q, A43T, T45A and the addition of A27, P28, A29 and E30 for AniA , and I133V, V146A, D206G, T208V for NorB .The acquisition of a working nitrite reductase for the putatively acquired 2022 bp sequence identified n = 103 exact, full-length matches, all of which were meningococcal. Thirty eight of these belonged to, or were closely related to, cc4821, including n = 29 cc4821 lineage 2c isolates from the current study with aniA allele 8 (n = 28) or aniA allele 733 (n = 1); five cc4821 carriage study isolates from the UK and Sweden that were not included in the current study ; and four clonal complex-unassigned isolates with <4 MLST alleles in common with ST-4821 . The latter four isolates each had 4 to 5 loci in common with ST-3200. The cc8 isolate descended of cc4821 was also identified. The remaining BLAST hits were for MenA ST-4 complex and MenA ST-5 complex isolates and two isolates with incomplete MLST profiles but matching cc5 at six loci.A BLAST search of all genomes (>2Mb) on the PubMLST mtrC (NEIS1634) allele 93 encoding an intact peptide.The six lineage 2c RoW anogenital isolates possessed High-resolution phylogenetic analyses have revolutionised our understanding of meningococcal population structures and our ability to identify and track existing and emerging strains and outbreaks on a global level , 32. SucA key finding of our study is the existence of four well defined lineages, versus the two that were previously identified , 6. LineNeisseria database , exclusively belonged to a distinct cluster (the RoW cluster) within lineage 2c. Furthermore, whilst PorA subtype P1.17\u20136,23 and closely-related subtypes were only associated with the RoW cluster among our genome panel, they also accounted for 14/27 PorA subtypes among non-Chinese, non-genomic isolate submissions on the PubMLST aniA) gene [aniA functionality, however, 14/17 lineage 2c RoW isolates, including the anogenital isolates, and 15/40 Chinese lineage 2c isolates shared a putative ancestral recombination involving partial sequences of the divergent aniA and norB genes along with the intergenic/promoter regions. Although the corresponding sequence was exclusively meningococcal on the PubMLST database, its initial origin was likely in MenA-associated lineages dating back to 1915. It is possible that the newly generated alleles or the newly acquired intergenic region (containing promoter regions) where many of the changes occurred, are more efficient than the existing ones. This, however, requires further investigation. Formal bioinformatic analyses are also required to confirm the origin of the putatively horizontally acquired sequence. Another possible explanation for this particular cluster alone being observed in anogenital sites is that Chinese anogenital meningococci from other lineage 2c clusters or the other cc4821 lineages may simply not be collected, sequenced, posted or identified as such on PubMLST. Interestingly, lineage 2c also included a rare isolate lacking an fhbp gene and therefore unable to express fHbp which has also previously been implicated in adaptation to the anogenital niche [mtrC that have been associated with cervical infection in gonococci and urogenitally adapted meningococci from uretheritis outbreaks among heterosexual males [More than a third (6/17) of the lineage 2c RoW cluster isolates were from an anogenital site among MSM. Although two groups of isolates (a group of three and a group of two) were from patients attending sexual health services that utilised common microbiology laboratories (at opposite ends of the UK), the corresponding isolates were distributed over time and within the population structure of the corresponding cluster. Furthermore, the second group also differed from one-another by serogroup. This, along with the finding of the recent msm carriage study in New York (USA) , may be iA) gene , 18. AmoNeisseria database (accessed 13th July 2020). None of the isolates from Europe , North America , South America , Australasia or Asia possessed the epidemic clone-associated PorA .gyrA alleles throughout the four cc4821 lineages and, with the exception of lineage 1, these were interspersed with isolates possessing susceptibility-associated (MICs \u22640.03 mg/L) WT alleles. A small number of well distributed isolates also possessed mutant parC alleles that may further increase fluoroquinolone MICs. Notably, the RoW cluster does not possess a mutant gyrA, however, it may be susceptible to stable acquisition of fluoroquinolone resistance given the situation in the broader clonal complex and the gradual emergence of fluoroquinolone resistance in Western countries [penA allele associated with reduced penicillin susceptibility (0.094 to 1 mg/L). Although diverse mutant penA alleles were found among Chinese isolates in each of the four lineages, they were relatively rare. As the present study constituted a genomic survey, we focussed on antibiotic resistance/reduced susceptibility determinants commonly associated with cc4821, rather than phenotypic MICs. Further work may include consideration of other potential resistance determinants such as multidrug efflux systems or beta lactamase, the latter of which has recently been reported among meningococci in North America and Europe [The widespread distribution of fluoroquinolone resistance in cc4821 is well documented . The preountries , 37\u201339. d Europe , 41.fhbp possessed a variant 2 or 3 gene and so the majority of isolates are potentially covered by Men-fHbp, though this would need confirming with the MeASurE assay [fhbp and one isolate lacked a function nhba. Such isolates and their corresponding strains have infrequently been reported before [Potential 4CMenB strain coverage of lineages 2a, 2b and 2c, including the RoW cluster was heavily reliant on NHBA. As the corresponding peptides are rare among existing MATS datasets there is a critical need for MATS analysis of isolates representing each of the main peptides. This may then be used to extend the current list of covered, not covered, and unpredictable isolates as defined by gMATS . PerhapsrE assay . Of concd before , 34, 42 In conclusion, cc4821 can now be considered in terms of four distinct lineages and the phylogeny herein can form a template with which to analyse genomes from prevailing cases. A small and distinct cluster of lineage 2c is responsible for the majority of cases outside of China and may have an affinity for anogenital sites. In light of emerging fluoroquinolone and penicillin resistance, uncertain strain coverage by subcapsular vaccines, and the repeated emergence of MenB cc4821 strains through capsule switching, there is an urgent need to test isolates representative of each lineage in the MATS and MeASurE assays.S1 Fig(DOCX)Click here for additional data file.S2 Fig(DOCX)Click here for additional data file.S3 Fig(DOCX)Click here for additional data file.S4 Fig(DOCX)Click here for additional data file.S5 Fig(DOCX)Click here for additional data file.S6 Fig(DOCX)Click here for additional data file.S7 Fig(DOCX)Click here for additional data file.S8 Fig(DOCX)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file."} +{"text": "Bioscience Reports (2019) 39, https://doi.org/10.1042/BSR20181016) would like to correct The authors of the original article above article \u201cThe effect of HMGB1 on the clinicopathological and prognostic features of cervical cancer\u201d ("} +{"text": "The proactive generation of anti-idiotypic antibodies (anti-IDs) against therapeutic antibodies with desirable properties is an important step in pre-clinical and clinical assay development supporting their bioanalytical programs. Here, we describe a robust platform to generate anti-IDs using rabbit single B cell sorting-culture and cloning technology by immunizing rabbits with therapeutic drug Fab fragment and sorting complementarity determining regions (CDRs) specific B cells using designed framework control as a negative gate to exclude non-CDRs-specific B cells. The supernatants of cultured B cells were subsequently screened for binding to drug-molecule by enzyme-linked immunosorbent assay and the positive hits of B cell lysates were selected for cloning of their immunoglobulin G (IgG) variable regions. The recombinant monoclonal anti-IDs generated with this method have high affinity and specificity with broad epitope coverage and different types. The recombinant anti-IDs were available for assay development to support pharmacokinetic (PK) and immunogenicity studies within 12 weeks from the start of rabbit immunization. Using this novel rapid and efficient in-house approach we have generated a large panel of anti-IDs against a series of 11 therapeutic antibody drugs and successfully applied them to the clinical assay development. All 24 unique anti-IDs identified in project E are listed and ranked by their actual Ag binding Rmax, [%] (highest to lowest).The binding response of Ab1 Fab against each protein A-captured Ab2 clone following Ag binding to Ab1 Fab was recorded. The actual Ag binding response (Ag\u2014Ab1 Fab) was used to determine the actual Ag binding Rmax [%] by dividing it with the theoretical Ag binding Rmax; Rmax = (DOCX)Click here for additional data file.S2 TableUnder similar protein A capturing level for each Ab2 clone the binding Rmax of Ag + Ab1 Fab as well as Ab1 Fab were recorded. The binding Rmax ratio between Ag + Ab1 Fab and Ab1 Fab was calculated as %. If the value was >10%, the clone was considered Ag and Ab1 complex specific epitope type. All 24 unique anti-IDs identified in project E are listed and ranked by their binding Rmax ratio ((DOCX)Click here for additional data file."} +{"text": "Investigators from the Hospital Dona Estef\u00e2nia, Escola Superior de Tecnologias e Sa\u00fade de Lisboa and Centro Hospitalar Psiqui\u00e1trico de Lisboa investigated 38 patients with continuous spike-wave of sleep (CSWS) syndrome. Investigators from the Hospital Dona Estef\u00e2nia, Escola Superior de Tecnologias e Sa\u00fade de Lisboa and Centro Hospitalar Psiqui\u00e1trico de Lisboa investigated 38 patients with continuous spike-wave of sleep (CSWS) syndrome. A 10-20 montage long-term ambulatory electroencephalography (aEEG) and a wearable aEEG with two bipolar EEG channels simultaneously validate the latter's usefulness to quantify the spike index (SI) to analyze the EEG response to the treatments. The SI was digitally calculated every 10 minutes by semiautomatic template-match-spike search analysis compared to both aEEG. They also compared daily SI stability and monthly SI stability (N=10) in individual patients and employed the SI obtained from averaging the maximum SI for each of the first four sleep cycles (avSI) for comparison to reduce variability. The wearable aEEG examinations were repeated up to 8 times per patient. The response to therapy using corticosteroids (n=10), sulthiame (n=7), and the ketogenic diet (n=3) was assessed by comparison of the avSI after therapy. As a result, corticosteroids most effectively reduced the SI among therapies, although sizeable individual variability in both the amount and time of onset of clinical responses. In conclusion, the wearable aEEG with two bipolar EEG channels, which are easily attached and tolerable for children, can provide accurate SI quantification repeatedly in clinical settings. [COMMENTARY. CSWS syndrome, also known as encephalopathy related to status epilepticus during slow sleep (ESES), is one of the most common epileptic encephalopathies of childhood . In thisThe author has declared that no competing interests exist."} +{"text": "Most existing ageism scales are designed to measure younger people\u2019s age attitudes, often asking respondents to affirm or reject stereotypes of older people. The Personal Beliefs about Aging (PBA) Scale gathers information about the degree to which respondents, regardless of age, value age diversity on their campus. Preliminary findings from a 2019-2020 University of Massachusetts system-wide electronic survey that yielded 2,563 responses across 3 (of 5) campuses indicate that most faculty (83%), staff (84%), and students (72%) perceived ageism as a serious problem in society; however far fewer considered ageism as a serious problem on their own campus, with students (20%) perceiving campus ageism to an even lesser extent than faculty (39%) and staff (36%). Part of a symposium sponsored by Age-Friendly University (AFU) Interest Group."} +{"text": "Differences in prefrontal cortex (PFC) control of walking in older age likely arise from changes in neural capacity and compensation. PFC activation by changes in oxygenated hemoglobin from functional near infra-red spectroscopy was examined in 29 older adults (mean age=76). Tasks included standing with cognitive challenge and walking with and without cognitive challenge on even and uneven surfaces. Three PFC activation-performance patterns were identified using K-means clustering: 1) low activation during walking tasks and high activation during standing cognitive task, with the best performance in terms of walking speed and cognitive performance (n=10); 2) low activation on all tasks, with the lowest performance (n=15); 3) high activation during walking and low activation during cognitive, with intermediate performance (n=5). Associations of patterns with cognitive function and structural neuroimaging were explored, with results informing interpretation of functional changes of PFC during aging process, including compensatory mechanisms for primary network impairment."} +{"text": "End Stage Renal Disease (ESRD) conveys high symptom burden, multimorbidity and the greater likelihood of hospital death than other serious illnesses. Increases in people with ESRD occurred most sharply among adults age 75+. Despite high mortality risk, few with ESRD consider end-of-life preferences or discuss with a physician. The purpose of this study was to explore the nature of advance care planning in ESRD. The study utilized mixed methods and both qualitative and quantitative data was collected during in-depth chairside interviews with 31 people while they were on hemodialysis. Participants ranged in age from 29-85; Mage=60; (N=13 ) were not considering a transplant while (N=9[69%]) of those under 60 had a failed transplant and were again on the waiting list. Although the majority of participants had a health care proxy (N=27[87%]), more who were >60 had a proxy who knew their wishes (N=14[78%]) compared with (N=9[69%]) who were 60 compared with none <60. The qualitative data illuminated these age-differentiated responses in themes: Older age and (1) Multimorbidity; (2) Frequency/intensity of hospitalization; (3) Diminished hope of transplantation; and (4) More acute death awareness. The need for disease-specific advance care planning\u2014with hopes and expectations about transplant--and attention to the influence of age and decline cannot be overstated."} +{"text": "This study examined factors affecting the feelings of loneliness among older Korean Americans. Data were drawn from a survey with older Korean Americans aged 60 or over in five states , conducted during 2017\u22122018. In hierarchical multiple regression models, loneliness was regressed on five blocks of variables: (1) demographic/health ; (2) immigration-related ; (3) social engagement ; (4) negative family interactions; and (5) interactions of negative family interactions with social engagement variables. A significant interaction was found in the relationship between friend network and negative family interactions: the impact of negative family interactions on loneliness was buffered by friend network. Implications of findings were discussed regarding working with older immigrants with limited social networks."} +{"text": "This cross-sectional study examines the representation of darker skin phenotypes in tools used by students in preparation for medical licensure testing. Although medical curricula vary widely, preparation for individuals seeking their preferred residencies in the US often converges on QBanks.3 Skewed representation of skin phenotypes in UWorld thereby risks development of biases during a critical moment in medical education.3 In this analysis of skin phenotype, we hypothesized that darker skin is underrepresented in images from the UWorld Step 2 CK QBank.The UWorld Step 2 Clinical Knowledge (CK) Question Bank (QBank) leads among study tools for medical students preparing for the second US Medical Licensing Examination (USMLE).4 into light (1-4), medium (5-6), and dark (7+) categories (\u03ba\u2009=\u20090.96); the coders resolved disagreements through discussion. We further categorized these images using the organization provided by UWorld and denoted images depicting any of 36 characteristic dermatologic findings.5 We used \u03c72 analyses to test differences among image types and systems by skin phenotype (\u03b1\u2009<\u2009.05 threshold of significance) and Poisson logistic regression to analyze variation in counts of images of hallmark dermatologic findings by skin phenotype. Testing was 2-tailed and unpaired. We entered data into Google sheets (Google LLC) and completed analyses in RStudio, version 1.1.463 (RStudio PBC). The Yale University Institutional Review Board determined this study did not constitute human subjects research. This study followed the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) reporting guideline.We examined 3537 questions in the UWorld Step 2 CK QBank between May 28 and August 11, 2020, recording and describing 1251 photographs or illustrations of human skin. Two of us serving as coders (J.P.C. and R.J.) independently classified these images by skin tone, collapsing the 11-point Project on Ethnicity and Race in Latin America skin color paletteP\u2009<\u2009.001) (P\u2009<\u2009.001) outnumbered medium (n\u2009=\u200947) and dark (n\u2009=\u200947) skin phenotypes; the ratio of light to darker skin phenotypes was 12 to 1 , A. Dark\u2009<\u2009.001) , B. Derm\u2009<\u2009.001) , C. Char\u2009<\u2009.001) , D. At l6 Our study could not assess for updates made to the QBank following the conclusion of the capture period or draw conclusions regarding the US Medical Licensing Examination Step 2 CK. Nevertheless, our analysis suggests that increasing representation of varied skin tones in medical training materials may mitigate health disparities rather than reproduce them. Partnership with the Skin of Color Society, Brown Skin Matters, and Mind the Gap organizations may support these efforts.Our findings suggest that darker skin phenotypes are underrepresented in the UWorld Step 2 QBank, a popular study tool for medical students seeking US residencies. UWorld contributors across multiple specialties should broaden representation of diverse skin phenotypes through both patient photographs and designer illustrations to ensure that clinicians are able to identify pathologic traits in darker-skinned patients. In addition, demonstration of skin cancers across a range of phenotypes may improve treatment prognosis."} +{"text": "Scientific Reports 10.1038/srep14484, published online 5 October 2015Correction to: The Supplementary Information file that accompanies this Article contains an error where the reference list was erroneously omitted.The corrected Supplementary Information."} +{"text": "We have the great pleasure to present this special issue consisting of selectedpeer-reviewed articles that were presented at the 2nd international biological massspectrometry symposium held from October 26th to 27th, 2018 at the Shimadzu HQ in Kyoto.The symposium was organized by the biological mass spectrometry (BMS) division of theMass Spectrometry Society of Japan. The focus of the symposium was on \u201cUnderstandingLife by MS,\u201d and featured four invited keynote lectures delivered by leading expertsfrom different countries and eight topics by local researchers. There were over 80attendees at the symposium representing various disciplines who engaged in vigorousdiscussions and interactions. The topics delivered at the symposium are listed asfollows:Development of a reference method for the determinationof HbA2.\u00b7Cristian Arsene Deciphering the lipid language of insects with ambientmass spectrometry.\u00b7Joanne Yew Capillary electrophoresis for single-cell massspectrometry of the early developing embryo.\u00b7Peter Nemes On the building of the Skyline targeted mass specrometrysoftware.\u00b7Brendan MacLean Keynote lectures:Standard materials and measurement methods leading toprecise peptide and protein measurements.\u00b7Tomoya Kinumi A smart nano-machine that demolishes microtubulecytoskeletons in cells.\u00b7Tadayuki Ogawa Phosphoproteome and kinome profiling usingnanoLC-MS/MS.\u00b7Naoyuki Sugiyama Hyphenation of high-temperature capillary liquidchromatography with electrospray ionization massspectrometry: Technical challenges & futureprospects.\u00b7Lee Chuin Chen Structural analysis of biomolecules using HydrogenAttachment/abstraction Dissociation (HAD) and relatednovel radical induced dissociation techniques.\u00b7Hidenori Takahashi 13C-Metabolic flux analysis of the metabolismof cancer cells.\u00b7Fumio Matsuda Roles of mass spectrometry in protein science andengineering.\u00b7Susumu Uchiyama Native MS: mass spectrometry for characterizingnon-covalent molecular complexs.\u00b7Kentaro Ishii Regular oral sessions:In addition to the regular oral session, seventeen students and young researchers alsotook part in short 5-min oral presentations and poster sessions. One symposium researchaward and three student symposium awards were presented for outstanding presentationsduring the symposium. The award evaluation committee consisted of nine board members ofthe BMS division. The winners were announced at the award ceremony which was held duringthe banquet.Cutibacteriumacnes.MALDI-MS proteotyping of \u00b7Kanae Teramoto The winner of the Symposium Research Award is:Development of high-temperature liquid chromatographyESI-MS.\u00b7Norhiedayah Rosli (Univ. Yamanashi)Analysis of metabolic responses to paclitaxel treatmentin central carbon metabolism of cancer cells.\u00b7Haruhiko Maruyama (Osaka Univ.)Stability and structural changes of an antibody indifferent buffers.\u00b7Hiroaki Oyama (Osaka Univ.)The winners of the Student Symposium Award are:Finally, we wish to take this opportunity to thank all of the members of the organizingcommittee: Susumu Uchiyama (Osaka Univ.), Tomoya Kinumi (AIST), Fumio Matsuda (OsakaUniv.), Rika Miyake (Osaka Univ.), Mayuko Morita (Kyoto Pref. Univ. of Medicine), andall of the reviewers for their work. It is our hope that this special issue will beuseful as reference materials for mass spectrometrists and life science researchers."} +{"text": "Supplemental Video) illustrates the dramatic appearance of Mf in the anterior chamber by a slit lamp.A 59-year-old man from an onchocerciasis-endemic area in eastern Ghana was enrolled in a clinical trial to assess the impact of ivermectin on microfilariae (Mf) in the skin and eyes. He had palpable subcutaneous nodules consistent with onchocerciasis and a high Mf burden in skin snips (170/mg of skin). He reported mild generalized pruritus and blurry vision in both eyes. Visual acuity was 20/80 (OD) and 20/60 (OS). Both eyes had moderate cataracts and features consistent with prior anterior uveitis. Intraocular pressures were normal (12 mmHg OU). Atrophic changes in his optic nerves correlated well with visual field defects measured by frequency doubling technology and with retinal nerve fiber layer loss as assessed by optical coherence tomography. Slit lamp examination revealed high numbers of motile Mf in the anterior chambers of both eyes (100-OD and 50-OS) . A cell-Ocular Mf were still present but decreased in numbers and motility (40-OD and 20-OS) 3 months after ivermectin treatment (150 mg/kg); no Mf were detected in four skin snips taken from each iliac crest and calf at that time. Microfilariae were still absent in skin snips taken 6 months after treatment when ocular Mf counts were 20-OD and 15-OS.Onchocerca volvulus Mf were cleared from the eyes more slowly than from the skin after ivermectin treatment. The patient\u2019s vision did not improve after treatment presumably because of irreversible effects of onchocerciasis.Supplemental materials"} +{"text": "Scientific Reports 10.1038/srep36482, published online 02 November 2016Correction to: This Article contains an error in the Introduction.The text,\u201cOn the other hand, although LDCT could reduce mortality by 20%, the high false positive rate (96.4%) observed in the NLST calls for more accurate risk stratification among heavy smokers.\u201dshould read:\u201cOn the other hand, although LDCT could reduce mortality by 20%, the high number of false positives within the positive results (96.4%) observed in the NLST calls for more accurate risk stratification among heavy smokers.\u201d"} +{"text": "Hepatocellular carcinoma (HCC) is one of the most common human malignancies worldwide with very poor prognosis. Resistance to targeted therapeutic drugs such as sorafenib remains one of the major challenges in clinical treatment. In the present study, PARP1 was found to be highly expressed in human embryonic stem cells, but progressively decreased upon specified hepatic differentiation. Reactivation of PARP1 expression was also detected in HCC residual tumors after sorafenib treatment in xenograft mouse model, indicating the potential important roles of PARP1 in stem cell pluripotency and HCC sorafenib treatment resistance. Overexpression of PARP1 was frequently observed in HCC patients, and closely associated with poor clinical outcome. Treatment of Sorafenib induced activation of DNA damage repair signaling, which is highly active and essential for maintenance of stem cell pluripotency in HCC residual tumors. PARP inhibitor Olaparib extensively suppressed the DNA damage repair signaling, and significantly inhibited the global pluripotent transcriptional network. The repression of key pluripotent transcriptional factors and DNA damage repair signaling by Olaparib was mainly through CHD1L-mediated condensation of the chromatin structure at their promotor regions. The global reshaping of the pluripotent transcriptome by Olaparib might reinforce Sorafenib in eliminating HCC residual tumors and enhance therapeutic efficiency.The online version contains supplementary material available at 10.1186/s12943-021-01315-9. Hepatocellular carcinoma is one of the most common human malignancies worldwide with poor prognosis . CurrentIncreasing evidences suggested that the hierarchy of cancer stem cells and their differentiated progenies contributed substantially to the heterogeneous tumor and therapeutic failure , 5. To bhESCs were differentiated into human hepatocytes along hepatic lineages. The whole differentiation process was defined with four stages, cells from the four different developmental stages were collected for transcriptomic profiling. . Inhibition of PARP1 by Olaparib could reverse such process, and thus exhibited a \u201cprotection\u201d effect, leading to reduced digestion and consequently more uncut DNA retained for qPCR detection. After MNase treatment, the number of uncut DNA fragments near the promoter region of target genes were determined by qPCR in cells with or without Olaparib exposure Fig. i. The tuIn conclusion, we found Sorafenib treatment could retain resistant tumor cells characterized with elevated cancer stemness and activation of DNA damage repair signaling. PARP1, which is highly activated in embryonic stem cells and Sorafenib resistant cancer cells, might be responsible for the active transcription of the pluripotent transcriptional factors and DNA damage repair signaling through maintaining an \u201copen chromatin\u201d structure. PARP inhibitor Olaparib extensively suppressed the pluripotent transcriptome through condensation of the chromatin structure and might greatly reinforce Sorafenib in eliminating HCC further in the clinic Fig. l.Additional file 1: Figure S1. Identification of PARP1 as a potential therapeutic target in HCC.Additional file 2.Additional file 3: Table S1.Additional file 4: Table S2.Additional file 5: Figure S2. PARP inhibitor Olaparib inhibits tumorigenesis in HCCAdditional file 6: Figure S3. Olaparib significantly potentiated Sorafenib both in vitro and in vivoAdditional file 7: Figure S4. Olaparib extensively suppressed the DNA damage repair signaling potentially through chromatin remodeling protein CHD1LAdditional file 8: Figure S5. Olaparib might repress the key pluripotency transcriptional factors through condensation of chromatin structureAdditional file 9: Table S3."} +{"text": "Conexibacter (17%), Nocardioides (8%), Streptomyces (7%), Geodermatophilus (6%), Methylobacterium (5%), and Burkholderia (4%). MG-RAST assisted analysis also revealed functional annotation based on subsystem, carbohydrates sequence had 13.74%, clustering based subsystem 12.93%, amino acids and derivatives 10.30% coupled with other useful functional traits needed for plant growth and health.This dataset presents shotgun metagenomic sequencing of sunflower rhizosphere microbiome in Bloemhof, South Africa. Data were collected to decipher the structure and function in the sunflower microbial community. Illumina HiSeq platform using next generation sequencing of the DNA was carried out. The metagenome comprised 8,991,566 sequences totaling 1,607,022,279\u202fbp size and 66% GC content. The metagenome was deposited into the NCBI database and can be accessed with the SRA accession number SRR10418054. An online metagenome server (MG RAST) using the subsystem database revealed bacteria had the highest taxonomical representation with 98.47%, eukaryote at 1.23%, and archaea at 0.20%. The most abundant genera were the Specification table1\u2022The data provides information on the rhizosphere microbiome of sunflower\u2022The microorganisms inhabiting the rhizosphere serves as hotspot for active biomolecules and novel genes\u2022Understanding the rhizosphere microbiome is important for plant growth and health\u2022These data are valuable, and this offers the possibilities of identifying new genes which could be an impetus for solving hunger and agricultural sustainability2The dataset contains a raw sequence data obtained using shotgun metagenomic of sunflower rhizosphere microbiome. The data files in FASTQ format were deposited at the National Center for Biotechnology Information (NCBI) with SRA accession number SRR10418054. The data are presented in 3www.mrdnalab.com), Texas, USA, using the Illumina HiSeq platform. Qubit\u00ae dsDNA HS Assay Kit (Life Technologies) was used to determine the initial concentration of DNA. Library preparation was done using the Nextera DNA Flex library preparation kit (Illumina) according to the manufacturer's guidelines. In brief, 50\u202fng of DNA were used for library preparation. After DNA fragmentation Illumina sequencing adapters were added and products amplified using six cycles of PCR during which unique indices were added. After library amplification, their concentration was estimated using the Qubit\u00ae dsDNA HS Assay Kit (Life Technologies), while the average library fragment size was measured using the Agilent 2100 Bioanalyzer (Agilent Technologies). Libraries were then pooled in equimolar ratios of 0.7\u202fnM and sequenced paired-end for 300 cycles using the NovaSeq 6000 platform (Illumina).Soil samples were obtained from sunflower rhizosphere soils in Bloemhof, South Africa (26.296138S: 26.972175E) and the DNA samples were extracted using the PowerSoil\u00ae isolation kit following the instructions of the manufacture. DNA concentration and purity were determined using a NanoDrop Lite Spectrophotometer . Extracted DNA was sent for metagenome shotgun sequencing to the Molecular Research Laboratory (www.mg-rast.org) was used for the quality control of the raw metagenome sequences The online metagenomic rapid annotation server MG-RAST failed to pass the quality control pipeline. Of those, dereplication identified 1046,824 sequences as artificial duplicate reads. Of the sequences that passed quality control, 17,387 sequences (0.24%) contain ribosomal RNA genes, 3722,027 sequences (52.74%) contain predicted proteins with known functions, and 3317,360 sequences (47.01%) contain predicted proteins with unknown function.The authors declare that they have no conflict of interest, either financial or commercial wise."} +{"text": "We studied whether vitamin D repletion could correct aberrant adipose tissue and muscle metabolism in a mouse model of CKD-associated cachexia. Intraperitoneal administration of 25(OH)D3 and 1,25(OH)2D3 (75\u00a0\u03bcg/kg/day and 60\u00a0ng/kg/day respectively for 6\u00a0weeks) normalized serum concentrations of 25(OH)D3 and 1,25(OH)2D3 in CKD mice. Vitamin D repletion stimulated appetite, normalized weight gain, and improved fat and lean mass content in CKD mice. Vitamin D supplementation attenuated expression of key molecules involved in adipose tissue browning and ameliorated expression of thermogenic genes in adipose tissue and skeletal muscle in CKD mice. Furthermore, repletion of vitamin D improved skeletal muscle fiber size and in vivo muscle function, normalized muscle collagen content and attenuated muscle fat infiltration as well as pathogenetic molecular pathways related to muscle mass regulation in CKD mice. RNAseq analysis was performed on the gastrocnemius muscle. Ingenuity Pathway Analysis revealed that the top 12 differentially expressed genes in CKD were correlated with impaired muscle and neuron regeneration, enhanced muscle thermogenesis and fibrosis. Importantly, vitamin D repletion normalized the expression of those 12 genes in CKD mice. Vitamin D repletion may be an effective therapeutic strategy for adipose tissue browning and muscle wasting in CKD patients.Patients with chronic kidney disease (CKD) are often 25(OH)D Current therapies focus on palliation, but calorie supplementation alone is not successful in treating CKD-associated cachexia3. Brown adipocytes and beige adipocytes, which reside within white adipose tissue (WAT), significantly contribute to whole body energy expenditure4. Beige adipocytes respond to cold stimulation in a process described as WAT browning5. We and others have demonstrated the presence of WAT browning in CKD mice5 amongst other animal disease models of cachexia as well as in patients with cachexia9. We also demonstrated WAT browning in a mouse model of cystinosis, a genetic cause of CKD11.Chronic kidney disease (CKD)-associated cachexia is a complex metabolic disorder that consists of anorexia, weight loss, loss of adipose tissue and muscle mass as well as hypermetabolism14. Vitamin D insufficiency12 may be an important cause of CKD-associated cachexia. Vitamin D influences myogenesis and muscle function16. Furthermore, vitamin D insufficiency has been correlated with reduced muscle size and strength in the general population. Supplementation of vitamin D was associated with increased muscle size and strength in patients on hemodialysis17. Vitamin D receptor (VDR) is expressed in skeletal muscle of mice and regulates uptake of 25-hydroxyvitamin D3 in myofibers18. Vitamin D exerts its effects via genomic and non-genomic, membrane-associated rapid response actions to regulate the function in muscle20. 25-hydroxyvitamin D3 could also directly exert paracrine and autocrine effects on muscle15. Low serum concentrations of 25-hydroxyvitamin D3 are an independent risk factor for decreased muscle function in elderly patients on dialysis21. Adipose tissue is an important storage site for vitamin D. Vitamin D insufficiency has been associated with aberrant adipogenesis22. There are no published data that report the direct impact of vitamin D repletion on muscle mass and adipose tissue metabolism in CKD mice. Thus, the purpose of this study was to investigate the impact of vitamin D repletion in a mouse model of CKD-associated cachexia.CKD patients have a high prevalence of 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 insufficiencySerum concentrations of 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 were significantly decreased in CKD mice compared to sham and skeletal muscle thermogenic genes were significantly increased in CKD mice were downregulated in CKD versus sham mice and decreasing expression of negative regulators of muscle mass as well as decreasing muscle protein contents of inflammatory cytokines IL-1\u03b2, IL-6 and TNF of CKD patients showed considerable variability, which may be influenced by degree of renal dysfunction, hyperparathyroidism, inflammation and the dialysis procedure26. Several investigations have reported that non-dialysis CKD patients have REE which may be similar or even slightly lower than that of age- and gender-matched healthy controls 28. These results may reflect the variable presence of cachexia in the pre-dialysis CKD population. Ikizler et al. carefully studied ESRD patients on maintenance hemodialysis and showed higher REE during resting conditions which was further increased during hemodialysis, compared with healthy controls29. Wang et al. studied a sizable population of 251 peritoneal dialysis patients, who were classified by REE tertiles. They showed that patients in the upper and middle tertiles of REE showed a significantly higher risk of all-cause mortality compared with those patients in the lower REE tertile30. Their findings highlight the significant negative prognostic effect of increased REE in CKD patients on long-term outcomes.Hypermetabolism is a cardinal feature of cachexia in CKD31. Increased expression of adipose and muscle UCPs have been implicated in various diseased-associated cachexia33. UCPs are mitochondrial inner membrane proteins positioned in the same membrane as the ATPase, which is also a proton channel. UCPs and ATPase work in parallel with UCPs generating heat and ATP synthase generating ATP. Increased UCPs expression stimulates thermogenesis while inhibiting ATP synthesis31. Adipose tissue and muscle UCPs content was increased while adipose tissue and muscle ATP content was significantly decreased in CKD mice versus sham mice -1 is important for thermogenesis while UCP-2 and UCP-3 regulate mitochondrial energy metabolism\u03b1, Cidea, Prdm16 and Dio2) and elevated skeletal muscle gene expression involved in fatty acid oxidation , UCP-3 and UCP-2 may act in concert in the overall regulation of fatty acid oxidation and energy expenditure39. Vitamin D repletion attenuated increased expression of both adipose tissue and skeletal muscle thermogenic genes in CKD mice . 1,25-dihydroxyvitamin D3 inhibits NF\u03baB activation. Interaction of MyD88 and Traf6 is critical for Tlr2-mediated transactivation of NF\u03baB and subsequent pro-inflammatory response45. We showed that repletion of vitamin D normalized inguinal WAT expression and protein content and protein content of inflammatory cytokines involved in promoting beige adipogenesis in CKD mice measured directly with 51Cr-EDTA, but a significant decline in estimated GFR using an estimation formula employing serum creatinine. Serum creatinine increased significantly by a mean by 18\u00a0mmol/l despite no significant change in measured GFR, implying an increase in muscle mass in these patients51. 1,25-dihydroxyvitamin D3 increased myogenic gene expression and inhibited myostatin expression, a negative regulator of muscle mass in C2C12 cells52.We studied the impact of vitamin D repletion on muscle fiber morphology and in vivo muscle function in CKD mice. To study muscle morphology, we chose to study two muscles known to have different fiber type compositions, namely soleus and tibialis anterior muscle. Soleus is a slow oxidative muscle while tibialis anterior is a fast glycolytic muscle55. Muscle adipose tissue may release pro-inflammatory cytokines within the muscle and impair the local muscle environment, impair blood flow or increase the rate of lipolysis within skeletal muscle resulting in an increased concentration of glucose within the skeletal muscle itself followed by insulin resistance57. We evaluated the effects of vitamin D repletion on muscular fat infiltration content in CKD mice. Our results clearly demonstrated that vitamin D supplementation attenuated muscle fat infiltration in CKD mice , proteolysis and inflammatory cytokines D3 , 1,25(OH)2D3 or vehicle control (ethylene glycol) using subcutaneous osmotic Alzet 2006 pump for 6\u00a0weeks. Oxygen consumption (VO2) was measured by using Oxymax indirect calorimetry (Columbus Instrument) and lean mass and fat content of mice were analyzed by quantitative magnetic resonance method 6. Muscle function (grip strength and rotarod activity) in mice was assessed using grip strength meter and rotarod performance test , respectively6.The study was in compliance with the approved animal protocol by the Institutional Animal Care and Use Committee (IACUC) at the University of California, San Diego. Schematic study design is listed 2D3, parathyroid hormone (PTH) and vitamin D binding protein (VDBP) were analyzed DAdenosine triphosphate (ATP) and uncoupling protein (UCP) contents in tissue homogenates were measured. Protein contents of pro-inflammatory cytokines in adipose and muscle tissue lysate were quantified 81.Fresh gastrocnemius muscles were preserved in isopentane/liquid nitrogen. Dissected muscle samples were incubated with Oil Red O . Detailed procedures for Oil Red O staining were in accordance with published protocolMuscle fibrotic and anti-fibrotic gene expression was analyzed . Detailed information for a total of 84 genes is listed in Supplemental Table www.bgi.com). The raw RNAseq data were filtered into clean reads, followed by mapping to the mouse reference genome using HISAT. The gene expression level for each sample was analyzed using RSEM quantification tool. Based on the gene expression level, differentially expressed genes (DEG) between CKD and sham mice were identified using DESeq2 algorithms. Biological function analysis of the DEG was enriched by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEEG) pathway. To identify pathways related to phenotypic differences of the muscle between CKD and sham mice, DEG between CKD and sham mice were analyzed through the use of IPA 82.We performed 5/6 nephrectomy and sham operation in 12-month old male C57BL/6\u00a0J mice. Total gastrocnemius muscle RNA was isolated in CKD and sham mice (3 mice in each group) using Trizol (Life Technology) followed by RNeasy mini kit (Qiagen) for further purification. The extracted muscle RNA samples were analyzed using Agilent 2100 Bioanalyzer (Agilent RNA 6000 Nano Kit). Samples were used to construct cDNA libraries (Illumina) and sequenced through an Illumina HiSeq2000 platform at BGI Hong Kong and reverse-transcribed with SuperScript III Reverse Transcriptase (Invitrogen). Quantitative real-time RT-PCR of target genes were performed using KAPA SYBR FAST qPCR kit (KAPA Biosystems). Expression levels were calculated according to the relative 2p value less than 0.05 was considered significant. Statistical analyses were performed using SPSS software version 16.0 for Macintosh.Continuous variables are expressed as mean\u2009\u00b1\u2009S.E.M. We assessed the statistical significance of differences between groups using two-sample t-tests. All tests were two-sided. A Supplementary information"} +{"text": "The average fungal population was 4.9 \u00d7 105 cfu/g with an average of pH 5.3 and 10.7%, respectively. In the present study, 131 fungal isolates were isolated and characterized based on macroscopic and microscopic characteristics and were grouped into 44 representative fungal strains. Based on results of morphological characteristics and ITS gene sequencing, 44 fungal strains were grouped into three phyla represented by Ascomycota (48%), Mucoromycota (38%), and Basidiomycota (14%). Taxonomical keys to species level was illustrated on the basis of morphological characteristics and ITS gene sequencing, aligned to the fungal database of NCBI GenBank, which showed seven genera with 16 species represented by Mucor circinelloides (20%), Aspergillus sydowii (11%), Penicillium chrysogenum (11%), Bjerkandera adusta (11%), Penicillium citrinum (7%), Rhizopus oryzae (7%), Aspergillus niger (5%), Aspergillus flavus (5%), Mucor indicus (5%) Rhizopus microsporus (5%), Rhizopus delemar (2%), Aspergillus versicolor (2%), Penicillium oxalicum (2%), Penicillium polonicum (2%), Trametes hirsuta (2%), and Cladosporium parahalotolerans (2%). The highest Shannon diversity index H was recorded in marcha of Sikkim (H: 1.74) and the lowest in hamei of Manipur (H: 0.69). Fungal species present in these amylolytic starters are morphologically, ecologically and phylogenetically diverse and showed high diversity within the community.Filamentous fungi are important organisms in traditionally prepared amylase and alcohol-producing dry starters in India. We collected 40 diverse types of amylase and alcohol-producing starters from eight states in North East India viz. The moisture content of the samples was estimated by a moisture analyzer .\u20134, 10\u20135, 10\u20136, and 10\u20137) was poured onto malt extract agar and potato dextrose agar with an addition of antibiotics (1% streptomycin) to suppress the growth of bacteria, and plates were then incubated under 28\u00b0C and observed for the appearance of colonies for up to 1 week. The colonies that appeared on plates were counted as a colony forming unit (cfu/g) on the dry weight of starters. Colonies were selected on the basis of macroscopic and microscopic characteristics. Selected filamentous molds were sub-cultured on new plates and purified and stored on slants at 4\u00b0C for further studies.Each dry sample starter was taken from the desiccator, then crushed coarsely by sterile spatula and 10 g of the crushed powered sample was homogenized with 90 mL of 0.85% physiological saline in a stomacher lab blender 40 for 2 min to obtain serial dilutions. One milliliter of each diluted sample , and microscopically by observation of fruiting bodies using a stereomicroscope, and the vegetative and asexual stages were observed by a DE/Axio Imager A1 microscope after staining freshly grown mycelia stained with cotton blue in MEA plates . FilamenThe genomic DNA was extracted from mold cultures following the methods of Polymerase chain reactions (PCR) of the internal transcribed spacer (ITS) region of filamentous molds was amplified using the primer ITS1 (5\u2032-TCCGTAGGTGAACCTGCGG-3\u2032) and ITS4 (5\u2032-TCCTCCGCTTATTGATATGC-3\u2032) . PCR reaThe amplified PCR products were purified using PEG (polyethylene glycol)-NaCl (sodium chloride) and precipitation solution with the addition of 0.6 volumes of 20% PEG-NaCl to the final volume of the PCR products . The mix\u20131 bovine serum albumin (BSA) and 0.04 U [\u03bcL]\u20131 tTaq DNA polymerase on a thermal cycler equipped with a heated lid. The thermal program included initial denaturation, enzyme activation at 95\u00b0C (6\u201310 min) followed by 35 cycles to complete the step and one cycle at 72\u00b0C (10 min). The amplified products were sequenced by an automated DNA Analyzer . These high-quality, double-stranded sequence data were analyzed with the help of the BLASTn program and multiple sequence alignment.PCR-amplified products had been sequenced in a forward and reverse direction using ITS1 primer (5\u2032-TCCGTAGGTGAACCTGCGG-3\u2032) and ITS4 primer (5\u2032-TCCTCCGCTTATTGATATGC-3\u2032), respectively, as per the method described by 1 using the Basic Local Alignment Search Tool for nucleotides (BLASTn) on the NCBI website and were edited using software ChromasPro version 1.34. Sequences were compared with sequence entries in the GenBank of NCBI website . For phy website .Percentages of frequency and relative density of fungal species in samples were determined as per the method described by Relative Density (%) was calculated by the equation:Diversity indexes of filamentous molds in samples were calculated by species richness (R), Shannon\u2019s diversity index (H), and species evenness (E) using PAMK396469\u2013MK396484, MK396486\u2013MK396500, MK778442\u2013MK778449, and MK796041\u2013MK796045.The sequences obtained in this study were deposited at the GenBank-NCBI database under accession numbers: 5 cfu/g collected from eight states of North East India . Mucor, Rhizopus, Aspergillus, Penicillium, and Cladosporium and a few unidentified basidiomycetes fungi were tentatively identified on the basis of detailed morphological characters using the taxonomical keys described by We isolated 131 total fungal isolates from 40 different samples of traditionally prepared dry starters in dawdim, and Basidiomycota (20%) in chowan, dawdim, and thiat, respectively. Phyla Ascomycota and Mucoromycota were present in all starters, whereas Basidiomycota was present only in marcha, thiat, chowan, and dawdim.Genomic DNA of each isolate of 44 representative fungal strains was extracted and PCR products were prepared for identification by ITS gene sequencing. DNA sequences of fungal isolates were assigned by comparison with those available in the GenBank of NCBI database using the ITS gene sequence (ITS1 and ITS4) based on the Basic Local Alignment Search Tool (BLAST) 2.0 program . The phyMucor circinelloides (20%), Aspergillus sydowii (11%), Penicillium chrysogenum (11%), Bjerkandera adusta (11%), Penicillium citrinum (7%), Rhizopus oryzae (7%), Aspergillus niger (5%), Aspergillus flavus (5%), Mucor indicus (5%) Rhizopus microsporus (5%), Rhizopus delemera (2%), Aspergillus versicolor (2%), Penicillium oxalicum (2%), Penicillium polonicum (2%), Trametes hirsuta (2%), and Cladosporium parahalotolerans (2%) . Interestaxonomy .Aspergillus niger was colonized with khekhrii; a species from the Mucor circinelloides complex was observed with a high dominance in samples, whereas Trametes hirsuta was less diversified and observed only in thiat samples and the lowest in hamei from Manipur (H: 0.69). Species Evenness (E) values were 0.97 in marcha followed by humao from Assam and phut from Arunachal Pradesh. The Species Richness (R), values were recorded highest in marcha and khekhrii samples (Diversity indexes of filamentous molds of dry starters were characterized by species richness (R), Shannon\u2019s diversity index (H), and species evenness (E) . The Sha samples .Drinking of cereal-based mild to strong alcoholic beverages produced by traditionally prepared amylase and alcohol-producing starters has been a traditional food culture of the ethnic people from the North East states of India for centuries. Traditionally prepared dry starters have consortia of co-existed microbiota containing filamentous molds, yeasts, and bacteria and are crudely sub-cultured through a \u201cback-slopping\u201d process by traditional starter-makers , 2019, fmarcha of the internal transcribed spacer (ITS) region of 44 strains of filamentous fungi isolated from starters from North East India using the primers ITS1 and ITS4 and grouped into three phyla represented by Ascomycota (48%), Mucoromycota (38%), and Basidiomycota (14%). A similar type of phylum distribution was also reported earlier in a om Korea and daquom China . Seven gtaxonomy . Our earE method supporte Vietnam . In marcom Nepal .Aspergillus belonging to order Eurotials is a phenotypically polythetic genus and is widely distributed in the environment and in a khekhrii sample from Nagaland (A. flavus NKM-7). Though the distribution percentage was only 5%, the presence of A. flavus in samples of marcha and khekhrii is alarming. A. flavus is a saprotrophic with cosmopolitan distribution which include M. circinelloides, M. griseocyanus, M. janssenii, M. lusitanicus, M. ramosissimus, M. variicolumellatus, and M. velutinosus , thiat , chowan (Tripura), and dowdim (Mizoram). The probable entry of P. chrysogenum during traditional preparation may be from damp and moist rooms where preparation for such starters is usually done, since P. chrysogenum is also found in damp buildings and P. polonicum in marcha samples. P. oxalicum produces various enzymes and natural products and lowest in hamei samples of Manipur (H: 0.69) indicating higher fungal diversity in marcha samples of Sikkim as compared to starters of other states. The diversity index, which considers both the number of species as well as relative abundance of each species for evaluating diversity region obtained in this study were deposited at the GenBank-NCBI database 6S rRNA sequencing were deposited at GenBank-NCBI numbers: MK396469-MK396484, MK396486-MK396500, MK778442-MK778449, MK796041-MK796045.AA performed the experiments. JT supervised the experiments and finalized the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Oxalis pes-caprae instead of the heading nasturtium Tropaelolum majus. Please see the correct There is an error in Results are per 100g of wet tissue except total phenolics and oxalic acid, which are concentrations (mg/g) See Table 6 for sample locations."} +{"text": "Selective intrauterine fetal growth restriction (sIUGR) in monochorionic diamniotic twins, especially types 2&3 with abnormal umbilical artery Doppler, results in increased risk of fetal/perinatal mortality and postnatal disability. We investigate whether the hair metabolome profiles of neonates were associated with the pathophysiological differences across the different clinical forms of sIUGR in twins.Hair samples were collected at delivery from 10 pairs of type 1 sIUGR twins, 8 pairs of types 2&3 sIUGR twins, and 11 pairs of twins without sIUGR. The hair metabolome was characterized using gas chromatography-mass spectrometry.Our results demonstrated that the hair metabolite profiles of the different sIUGR subclinical forms were associated with the averaged fetal growth rate after 28\u2009weeks of gestation but not with birthweight. The hair profiles were capable of discriminating type2&3 sIUGR twins from twins without sIUGR. In particular, the metabolites 2-aminobutyric acid, cysteine, alanine, and tyrosine all displayed areas under the receiver operating characteristic curve were above 0.9. The metabolic pathway analysis highlighted the associations of sIUGR twins with abnormal umbilical artery flow with increased metabolites from a nutrient depletion pathway, glutathione metabolism, and nerve development.This study offers novel insight into the severity of intrauterine ischemia and hypoxia for T2&3 sIUGR twins, through evaluation of the neonatal hair metabolome. Unequal sharing of the placental and vascular communications between twins leads to a significantly higher risk of various complications in monochorionic (MC) twins compared to dichorionic (DC) twins occurs in 10\u201315% of MC twin pregnancies. The clinical classification of sIUGR is according to Doppler waveform of end-diastolic velocity in umbilical artery flow (UAF), diagnosed by ultrasound present in a cell, tissue, or organism. Although metabolomic studies of MCDA twins with sIUGR have occurred in cord plasma and placenta . A total of 142 MCDA twin pregnancies were enrolled in the cohort between September 2017 and December 2018. Participants with maternal chronic diseases, major fetal structural anomalies, other adverse twin pregnancy outcomes, and those lost to follow-up at birth were excluded from participation in this study was determined by calculating weekly average growth of estimated fetal weight (EFW) from 28 gestational weeks until delivery using linear interpolation displaying a single gestational sac via ultrasound by 10 gestational weeks and b) presenting with T-sign of the intertwin membrane and absent twin peak (lambda) sign via ultrasound between 11 and 14 gestational weeks. A final pathologic evaluation of the placental features after delivery confirmed monochronic twins participated in this study. The diagnostic criteria for sIUGR was defined by at least one of the twin\u2019s EFW being less than the 10th percentile for the corresponding gestational age and an intertwin EFW discordance greater than 25%. The sIUGR twins were then classified into type-1/2/3 sIUGR using Doppler velocimetry via ultrasound of the end-diastolic umbilical arteries as detailed in Fig.\u00a0Immediately after twin delivery, hair samples were cut 0.5\u2009cm proximal from the scalp of each twin with clean scissors and then stored at \u2212\u200920\u2009\u00b0C until the samples were processed. All hair samples underwent only a single freeze-thaw cycle before analysis.Hair sample preparation was conducted according to the published protocol by Sulek et al with minor modifications and methyl chloroformate derivatization was performed to make the compounds more volatile for GC-MS analysis, based on the protocol published by Smart et al with leave-one-out cross validation (LOOCV) was performed to compare the hair metabolome profiles between different twin groups via Metaboanalyst 3.0 package for R . Generalized estimating equation (GEE) modeling was implemented to identify hair metabolites correlated with birthweight and growth rate discordances within twin pairs and between twin pairs. Metabolic pathway activity was estimated using the Pathway Activity Profiling (PAPi) R-algorithm and respective GC retention time to an in-house MS library established using chemical standards. The remaining putative compounds were identified using a commercial NIST mass spectral library. The MassOmics XCMS R-based script was used to extract the relative concentration of the metabolites through the peak height of the most abundant fragmented ion mass. To improve quantitative robustness along with minimizing human and instrumental variability, the relative abundances of the identified compounds were normalized in the order of multiple internal standards , median centering-batch correction via nine QC samples (three QC samples per batch), and dried hair weight of the corresponding sample. Then, blank samples were used to subtract background contamination and any carryover from identified metabolites. Student\u2019s The MCDA twin pregnancies were organized into three maternal groups: A) sIUGR pregnancy with one of the umbilical cords observed as normal UAF type 1); B) sIUGR pregnancy with abnormal UAF types 2&3); C) Control twin without any adverse pregnancy condition. Each pregnancy group (except control pregancy which was considered as a single group) was further divided into a larger and a smaller twin pair resulting in a total of five twin fetal groups with the following acronyms: 1) Type 1 sIUGR smaller twin (T1 sIUGR-S); 2) Type 1 sIUGR larger twin (T1 sIUGR-L); 3) Type 2 & type 3 sIUGR smaller twin (T2&3 sIUGR-S); 4) Type 2 & type 3 sIUGR larger twin (T2&3 sIUGR-L)); 5) Control twin. The unequal placental sharing, discordant velamentous cord insertion, large arterioarterial anastomoses, and abnormal umbilical artery flow of T1&2&3 sIUGR are shown in Fig.\u00a0; B) sIUG&3; C) Cop-value\u2009=\u20090.001) and control MCDA pregnancies . This was due to the fact that sIUGR pregnancies with abnormal UAF were delivered by cesarean section prior to 32\u2009weeks\u2019 gestation to prevent stillbirth.Maternal characteristics of T1 sIUGR, T2&3 sIUGR, and normal MCDA pregnancies were listed in Supplementary Table The comparison of perinatal outcomes is described in Supplementary Table https://www.nist.gov/nist-research-library) with the inter-assay coefficient of variation in QC samples ranging from 1.1 to 29.6% and between twin pairs (comparison between each co-twin pairs), a GEE regression model was performed, which also accounted for the individual (twin) and shared factors in this study. In terms of birthweight is associated with high risk of fetal and perinatal mortality for both twins, or long-term complications following exposure to ischemia and hypoxia in the uterus, especially in the smaller twin of the neonatal hair metabolome. Supplementary Figure 3. A generalized estimating equation to correlate the hair metabolites associated with birthweight discordance within and between MCDA twin pairs between T1 sIUGR, T2&3 sIUGR, and control twins. Supplementary Figure 4. A generalized estimating equation to correlate the hair metabolites associated with the growth rate discordance within and between MCDA twin pairs between T1 sIUGR, T2&3 sIUGR, and control twins. Supplementary Figure 5. Intra-observer variability in the measurement of fetal growth rate. Supplementary Figure 6. The inter-observer variability (a), intra-observer A variability (b) and intra-observer B variability (c) for determining gestational age. Supplementary Figure 7. PCA analysis of hair metabolite profile (n\u2009=\u20093 per group) stored at various temperatures over six months."} +{"text": "Periodic climatic oscillations and species dispersal during the postglacial period are two important causes of plant assemblage and distribution on the Qinghai\u2010Tibet Plateau (QTP). To improve our understanding of the bio\u2010geological histories of shrub communities on the QTP, we tested two hypotheses. First, the intensity of climatic oscillations played a filtering role during community structuring. Second, species dispersal during the postglacial period contributed to the recovery of species and phylogenetic diversity and the emergence of phylogenetic overdispersion. To test these hypotheses, we investigated and compared the shrub communities in the alpine and desert habitats of the northeastern QTP. Notably, we observed higher levels of species and phylogenetic diversity in the alpine habitat than in the desert habitat, leading to phylogenetic overdispersion in the alpine shrub communities versus phylogenetic clustering in the desert shrub communities. This phylogenetic overdispersion increased with greater climate anomalies. These results suggest that (a) although climate anomalies strongly affect shrub communities, these phenomena do not act as a filter for shrub community structuring, and (b) species dispersal increases phylogenetic diversity and overdispersion in a community. Moreover, our investigation of the phylogenetic community composition revealed a larger number of plant clades in the alpine shrub communities than in the desert shrub communities, which provided insights into plant clade\u2010level differences in the phylogenetic structures of alpine and desert shrub communities in the northeastern QTP. In this manuscript, we studied the phylogenetic community composition of shrub in the northeastern Qinghai\u2010Tibetan Plateau and found that existing significant difference in phylogenetic community composition of shrub communities from different habitats , which interpreted phylogenetic community structure from perspective of phylogenetic community composition, enhanced our knowledge of shrub, and helped us to obtain further interpretation for present\u2010day community structure from historical processes. Additionally, we generated principal coordinates of the phylogenetic structure (PCPS) , which has an average elevation, mean annual temperature range, and mean annual precipitation range of >\u00a03,000\u00a0m above sea level, \u22123.7 to 6.0\u00b0C, and 17\u2013776\u00a0mm, respectively anomaly to represent the climatic oscillations and mean(MNTDnull) represent the mean MPD and MNTD expected by chance, for which species were drawn 999 times from the species pool while using the same probability and maintaining the same species richness; and SD(MPDnull) and SD(MNTDnull) are the standard deviations of the MPD and MNTD expected by chance. A positive NRI or NTI indicates that a community comprises closely related species and has a clustered phylogenetic structure. Conversely, a negative NRI or NTI indicates that the species in a community are more distantly related than would be expected by chance and that the phylogenetic community structure is overdispersed. An NRI or NTI close to 0 or that differs nonsignificantly from 0 indicates no significant phylogenetic community structure Webb,\u00a0.Picante between the two types of shrub communities , soil water content (SWC), and coverage and no correlations with the other environmental factors Table\u00a0.3.3Pearson's correlation analysis revealed a stronger correlation between the NRI and PCPS I than between the NRI and PCPS II Table\u00a0. Signifi44.1We observed significant differences in the phylogenetic diversity and phylogenetic community structures when we compared shrubs between the alpine and desert habitats Figures\u00a0 and 3. SWe determined that the paleoclimatic anomaly representing the current MAT to the LGM\u2010MAT had the strongest effect on the shrub communities Figure\u00a0. This obSpecies dispersal in the postglacial period has had important effects on the plant community ; Data curation (lead); Formal analysis (lead); Funding acquisition (supporting); Investigation (lead); Methodology (lead); Project administration (supporting); Resources (lead); Software (lead); Supervision (lead); Validation (lead); Visualization (lead); Writing\u2010original draft (lead); Writing\u2010review & editing (lead). Lucun Yang: Conceptualization (supporting); Data curation (supporting); Formal analysis (supporting); Funding acquisition (supporting); Investigation (supporting); Methodology (supporting); Project administration (supporting); Resources (supporting); Software (supporting); Supervision (supporting); Validation (supporting); Visualization (supporting); Writing\u2010original draft (supporting); Writing\u2010review & editing (supporting). Xiuqing Nie: Conceptualization (supporting); Data curation (supporting); Formal analysis (supporting); Funding acquisition (supporting); Investigation (supporting); Methodology (supporting); Project administration (supporting); Resources (supporting); Software (supporting); Supervision (supporting); Validation (supporting); Visualization (supporting); Writing\u2010original draft (supporting); Writing\u2010review & editing (supporting). Changbin Li: Conceptualization (supporting); Data curation (supporting); Formal analysis (supporting); Funding acquisition (supporting); Investigation (supporting); Methodology (supporting); Project administration (supporting); Resources (supporting); Software (supporting); Supervision (supporting); Validation (supporting); Visualization (supporting); Writing\u2010original draft (supporting); Writing\u2010review & editing (supporting). Feng Xiong: Conceptualization (supporting); Data curation (supporting); Formal analysis (supporting); Funding acquisition (supporting); Investigation (supporting); Methodology (supporting); Project administration (supporting); Resources (supporting); Software (supporting); Supervision (supporting); Validation (supporting); Visualization (supporting); Writing\u2010original draft (supporting); Writing\u2010review & editing (supporting). Lingling Wang: Conceptualization (supporting); Data curation (supporting); Formal analysis (supporting); Funding acquisition (supporting); Investigation (supporting); Methodology (supporting); Project administration (supporting); Resources (supporting); Software (supporting); Supervision (supporting); Validation (supporting); Visualization (supporting); Writing\u2010original draft (supporting); Writing\u2010review & editing (supporting). Guoying Zhou: Conceptualization ; Data curation ; Formal analysis ; Funding acquisition (lead); Investigation (lead); Methodology ; Project administration (lead); Resources (lead); Software (supporting); Supervision (lead); Validation ; Visualization ; Writing\u2010original draft ; Writing\u2010review & editing .Fig S1Click here for additional data file.Fig S2Click here for additional data file.Fig S3Click here for additional data file.Table S1Click here for additional data file.Appendix S1Click here for additional data file."} +{"text": "Magnetococcales. These genomes contain nearly complete magnetosome gene clusters responsible for magnetosome biomineralization.Magnetotactic bacteria represent a valuable model system for the study of microbial biomineralization and magnetotaxis. Here, we report two metagenome-assembled genome sequences of uncultivated magnetotactic bacteria belonging to the order Magnetococcales. These genomes contain nearly complete magnetosome gene clusters responsible for magnetosome biomineralization.Magnetotactic bacteria represent a valuable model system for the study of microbial biomineralization and magnetotaxis. Here, we report two metagenome-assembled genome sequences of uncultivated magnetotactic bacteria belonging to the order Magnetococcales are often the dominant MTB group in nature are a diverse group of microorganisms that swim along the geomagnetic field lines, a behavior known as magnetotaxis or microbial magnetoreception . MTB affn nature , 3. Thisn nature or a subn nature near theacteria\u201d , 7 or Matococcia ) within Magnetococcales genome sequences isolated from freshwater sediments. Surface sediments were collected from East Lake in Hubei Province, China . MTB cells were magnetically enriched from 300\u2009ml of sediments using a double-ended open magnetic separation apparatus known as the \u201cMTB trap\u201d (Bacteria). Coverage information was determined using Bowtie 2 v2.3.4.3 shows the best hit to an uncultured magnetotactic coccus, MDA-1 (Magnetic coccus CS92 (X81182) genome sequences, designated DH2bin6 and DH2bin20, have been reconstructed here, both of which contain partial 16S rRNA genes (>900\u2009bp). An analysis of the 16S rRNA gene sequence of DH2bin6 using the online NCBI BLASTn nucleotide collection (nonredunant [nr]/nucleotide) database (dentity) , while D(X81182) . Genomes(X81182) . Nearly JAANAU000000000 and JAANAV000000000 (BioProject number PRJNA400260). The raw metagenomic read data have been deposited in the NCBI Sequence Read Archive under the accession number SRR11267947.These two genome sequences have been deposited in GenBank under the accession numbers"} +{"text": "This article provides data on the scanning tunnelling microscopy (STM), atomic force microscopy (AFM) and Kelvin probe force microscopy (KPFM) images of InAs(001) surface. Using the frequency-modulation (FM) method in AFM and KPFM, atomic resolution topography and contact potential difference (CPD) images of InAs(001) surface were obtained. The InAs(001) surface reconstruction images observed by STM and AFM are compared. The effect of AFM tip condition and tip-sample distance to AFM and KPFM imaging is verified by measuring frequency shift vs. tip-sample distance spectroscopy. This data article is related to the article entitled, \u201cKelvin prove force microscopy and its application\u201d [1]. The InAs samples are commercially available InAs wafer (Wafer Tech) with a 200 nm thick InAs surface layer grown by MBE. The STM and AFM measurements were performed with Omicron VT-AFM/STM system with an Omicron KPFM module. In order to obtain high resolution, AFM and KPFM imaging were performed in FM mode using super sharp silicon AFM tip (Nanosensors). The Si AFM tip was coated with 2\u20133 nm of Cr to make the tip conductive for KPFM imaging. FM mode KPFM was performed with a cantilever with resonant frequency of approximately 270 kHz.All experiments were performed in an ultrahigh vacuum chamber with a base pressure of 2\u00a0\u00d7\u00a010"} +{"text": "Otocyon megalotis, is the only member of its genus and is thought to occupy a basal position within the dog family. These factors can lead to challenges in complete mitochondrial reconstructions and accurate phylogenetic positioning. Here, we present the first complete mitochondrial genome of the bat-eared fox recovered using shotgun sequencing and iterative mapping to three distantly related species. Phylogenetic analyses placed the bat-eared fox basal in the Canidae family within the clade including true foxes (Vulpes) and the raccoon dog (Nyctereutes) with high support values. This position is in good agreement with previously published results based on short fragments of mitochondrial and nuclear genes, therefore adding more support to the basal positioning of the bat-eared fox within Canidae.The bat-eared fox, Otocyon megalotis) is a small member of the Canidae family and the only species of the genus Otocyon. It occurs in two allopatric populations across Africa (Clark Vulpes) and the Raccoon dog (Nyctereutes procyonoides) under pDNA was extracted using a Zymo genomic DNA clean and concentrator extraction kit, built into Nextera Illumina sequencing libraries and sequenced on an Illumina Nextseq 500 . We trimmed raw reads using Cutadapt v1.4 Martin , merged Canis lupus (Genbank accession KC461238.1), Vulpes vulpes (Genbank accession JN711443.1), and Urocyon littoralis (Genbank accession KP128962.1). Consensus sequences were called using a minimum read coverage of 10\u00d7 and a 75% base call threshold in Geneious v9.0.5 contains 16,431\u2009bp. The genomes produced were identical regardless of starting bait reference. Our bat-eared fox mitochondrial assembly contained all 13 protein-coding genes with expected open reading frames, 22 transfer RNA genes, and 2 ribosomal RNA genes found within a typical vertebrate mitochondrial genome.Otocyon was placed with high confidence (posterior probabilities\u2009>\u20090.99) as sister to the clade containing both the raccoon dog (Nyctereutes) and true foxes (Vulpes) . This re(Vulpes) ."} +{"text": "Forensic haplotype analysis of the male Y chromosome is currently used to establish the number of male donors in sexual assaults, the number of male bleeders in blood pattern analysis, and for ancestry correlation to genetic founder populations in biogeographic studies. In forensic laboratory applications, its primary use is for DNA profile generation with trace amounts of male DNA in the presence of excess female DNA . Our study supports the potential use of the Y chromosome in a \u201cdragnet\u201d approach (most haplotypes are unique) similar to that described by Kayser in 2017 for solving a cold case sex assault and homicide in The Netherlands. Our study also researched the potential for the identification of an ancestral Irish genetic \u201cfootprint\u201d linked to surname O\u2019Brien and identified multiple founder group origins in Ireland and England as well as three samples with the Dal Riata (a Gaelic overkingdom) ancestral haplotype. This study indicates correlation to ancestral Irish ancestry by haplotype but not conclusively to the O\u2019Brien surname. Genetic markers are used routinely for forensic applications and DNA databases . Y-STRs https://www.familytreedna.com/groups/ireland-heritage/about, accessed May 9, 2019). A second project researches the history of the Irish clans to identify founder populations and associated relatives many generations later. The goals of the Irish clan Y-DNA project are to connect genetic relatives and verify paternity and county of origin. Over 60 Irish clans are listed . A third project has easily accessible haplotype data for identification of an O\u2019Brien genetic footprint or common haplotype. The O\u2019Brien surname project is available at website https://www.familytreedna.com/groups/obrien/about/background, accessed October 10, 2018). This website consists of sampled individuals correlated by geography, Y-chromosome haplotype and single nucleotide polymorphisms (SNPs).There are several Irish ancestry projects related to biogeographical ancestry. Three major ones are described here. The Ireland Y-DNA Project has 8\u2009580 number break noted and offers both Y-chromosome testing and mitochondrial DNA testing to add to verification of historical information regarding ancestry. One hundred and eighty-one samples with surname O\u2019Brien are included in this project following the buccal swab spin protocol [\u00ae Yfiler\u00ae PCR Amplification kit (ThermoFisher Scientific) manufacturer\u2019s protocol on an Applied Biosystems PCR GeneAmp 9700 thermocycler (ThermoFisher Scientific). After PCR amplification, sample fragment separation and detection was performed on an Applied Biosystems 3130xl Prism\u00ae Genetic Analyzer (ThermoFisher Scientific). The results were then analyzed using GeneMarker software to score alleles at each genetic locus. The haplotypes were transferred to Excel software for data analysis. A pair-wise analysis was performed on all the samples within each population. The resulting data were assessed for a common genetic footprint between specific populations. When reporting new match criteria with Y-chromosome polymorphisms, the DNA Commission of the International Society of Forensic Genetics (ISFG) recommends three methods: counting method, likelihood ratio and Bayesian statistic [Participants in our study were selected based on surname and ancestry and were asked to fill out a questionnaire (Appendix I) to gather background information (with informed consent). Buccal samples were collected and processed for male DNA haplotypes. Sample collection was performed according to University of New Haven guidelines and after Institutional Review Board (IRB) approval. Following human sampling, DNA from cotton swabs was extracted using the QIAampprotocol . DNA exttatistic . Our stuTen O\u2019Brien\u2019s of Irish descent were able to be successfully sampled and analyzed . From thAfter the pair-wise analysis of the randomly sampled O\u2019Brien population was completed, trends were reviewed to determine if there were any similarities in males with varying locations of paternal origins and locations of origins of birth . In the https://www.familytreedna.com/groups/obrien/about/background. This is a large Irish population database that is being genotyped for common genetics based on a surname and is a larger dataset of independently collected and analyzed O\u2019Brien samples. The O\u2019Brien surname project has a common \u201cgenetic footprint\u201d of 13-24-29-17-14-(11-14)-13-11-11-13-11-15-12-19 (DYS389I-DYS390-DYS389II-DYS458-DYS19-DYS385-DYS393-DYS391-DYS439-DYS392-YGATAH-DYS437-DYS438-DYS448). Our samples represent Irish that have emigrated out of Ireland to the US. Our 10 samples did not share this exact O\u2019Brien surname \u201cgenetic footprint\u201d, but remnants of the footprint remained within the haplotype. Percent sharing of the O\u2019Brien surname alleles for our samples was: DYS389I (40%), DYS390 (30%), DYS389II (40%), DYS458 (30%), DYS19 (90%), DYS385 (80%), DYS393 (90%), DYS391 (60%), DYS439 (20%), DYS392 (60%), YGATAH (10%), DYS437 (70%), DYS438 (80%), DYS448 (40%). This allele frequency was then compared to 10 randomly sampled Caucasian with no O\u2019Brien surname connection. Percent sharing of the general Caucasian sampling with the O\u2019Brien surname alleles was: DYS389I (60%), DYS390 (30%), DYS389II (40%), DYS458 (70%), DYS19 (90%), DYS385 (50%), DYS393 (70%), DYS391 (30%), DYS439 (40%), DYS392 (60%), YGATAH (50%), DYS437 (50%), DYS438 (60%), DYS448 (50%). This information suggests that the most common alleles are more fixed in the overall population while the residual O\u2019 Brien surname \u201cgenetic footprint\u201d may be only loosely correlated after emigration.A potential relationship between surnames and Y-STR haplotype was demonstrated in a study conducted by Sykes and Irven . In thisOur best interpretation is that the Y-chromosome and O\u2019Brien surname is evolving over generations and once geographically unrestricted to Irish birthplace, only portions of the O\u2019Brien surname \u201cgenetic footprint\u201d remain. In fact, this is the strategy used for determining genetic relationships by paternal ancestry and is used routinely in evolutionary studies and family tree analyses. When comparing our O\u2019Brien surname samples to the Oxford Genetic Atlas Project (OGAP) data in the Campbell study for R1b haplotypes, three samples carry the Irish ancestral signature of the Dal Riata, two samples bear the Northern Isles signature, two samples are consistent with Northern England and one sample is consistent with Central England . The rem"} +{"text": "Fluorescence intensity (FI) was assessed using fluorescence spectroscopy and validated by fluorospectrophotometer and targeted liquid chromatography mass spectrometry (LC-MS/MS). Fluorescence intensity (FI) increased over time during machine perfusion in both groups (p<0.0001). This increase was similar for both groups (p = 0.83). No correlation, however, was found between FI and post-transplant outcomes, including day 5 or 7 serum creatinine , immediate graft function (p = 0.91), creatinine clearance and biopsy-proven rejection at one year . LC-MS/MS validation experiments of samples detected FMN in only one perfusate sample, whilst the majority of samples with the highest fluorescence remained negative. In the context of clinical kidney HMP, fluorescence spectroscopy unfortunately appears to be not specific and probably unsuitable for FMN. This study shows that FMN does not classify as a clinically relevant predictive biomarker of kidney graft function after transplantation.Hypothermic machine perfusion (HMP) provides preservation superior to cold storage and may allow for organ assessment prior to transplantation. Since flavin mononucleotide (FMN) in perfusate has been proposed as a biomarker of organ quality during HMP of donor livers, the aim of this study was to validate FMN as a biomarker for organ quality in the context of HMP preserved kidneys. Perfusate samples (n = 422) from the paired randomised controlled COPE-COMPARE-trial, comparing HMP with oxygenation (HMPO Hypothermic machine perfusion (HMP) is currently implemented to preserve kidney allografts from deceased donors. It has been demonstrated that HMP reduces the risk of delayed graft function (DGF) and improves graft survival \u20136. An adFlavin mononucleotide (FMN) is a cofactor for the mitochondrial membrane NADH:ubiquinone oxidoreductase enzyme (complex-I), in addition to a number of other proteins . Whilst 2) of deceased donor livers, suggesting that FMN release reflects a compromised energy status of the donor organ. The clinical relevance of this marker during liver perfusion was underpinned by the observation that FMN release during perfusion strongly associates with lactate clearance and coagulation factors at post-transplant days 1 and 2 .(DOCX)Click here for additional data file.S3 Table\u2020 Pearson correlation test was used for correlation between creatinine clearance and FI measured in perfusates taken during perfusion (P2). \u2021 Logistic regression analyses were used for association between FI and graft failure or rejection. With regards to rejection at one year, the numbers were too small to perform the analysis.Data are presented as correlation coefficient (r) or odds ratio with corresponding [95% Confidence Interval].(DOCX)Click here for additional data file.S4 Table\u2020 Spearman correlation test was used for correlation between creatinine clearance and FI from the delta perfusion (\u0394P) measured as (P3) perfusates taken at the end of perfusion\u2013(P1) perfusates taken at the beginning of perfusion.\u2021 Logistic regression analyses were used for association between FI and graft failure or rejection. With regards to rejection at one year, the numbers were too small to perform the analysis.Data are presented as correlation coefficient (r) or odds ratio with corresponding [95% Confidence Interval].(DOCX)Click here for additional data file.S1 AppendixFMN analysis in perfusate using targeted liquid chromatography mass spectrometry.(DOCX)Click here for additional data file.S2 Appendix(DOCX)Click here for additional data file.S1 Dataset(SAV)Click here for additional data file.S2 Dataset(PZFX)Click here for additional data file.S1 File(DOCX)Click here for additional data file."} +{"text": "Secondly, rod signals can enter cones through gap junctions. Finally, rods can synapse directly onto cone OFF bipolar cells.Light responses of rod photoreceptor cells traverse the retina through three pathways. The primary pathway involves synapses from rods to ON-type rod bipolar cells with OFF signals reaching retinal ganglion cells (RGCs) To analyze these pathways, we obtained whole cell recordings from OFF-type \u03b1 RGCs in mouse retinas while expressing channelrhodopsin-2 in rods and/or cones.2+ sensor, synaptotagmin 1 (Syt1), from cones abolished cone-driven optogenetic responses in RGCs. Rod-driven currents were not significantly reduced after isolating the secondary pathway by eliminating Syt1 and synaptotagmin 7 (Syt7) to block synaptic release from rods. Eliminating Syt1 from both rods and cones abolished responses to optogenetic stimulation. In Cx36 KO retinas lacking rod-cone gap junctions, optogenetic activation of rods evoked small and slow responses in most OFF RGCs suggesting rod signals reached them through an indirect pathway. Two OFF cells showed faster responses consistent with more direct input from cone OFF bipolar cells.Optogenetic stimulation of rods or cones evoked large fast currents in OFF RGCs. Blocking the primary rod pathway with L-AP4 and/or strychnine reduced rod-driven optogenetic currents in OFF RGCs by ~1/3. Blocking kainate receptors of OFF cone bipolar cells suppressed both rod- and cone-driven optogenetic currents in OFF RGCs. Inhibiting gap junctions between rods and cones with mecloflenamic acid or quinpirole reduced rod-driven responses in OFF RGCs. Eliminating the exocytotic CaThese data show that the secondary rod pathway supports robust inputs into OFF \u03b1 RGCs and suggests the tertiary pathway recruits both direct and indirect inputs. The primary OFF pathway can therefore be blocked with a glycine receptor antagonist strychnine. A second pathway emerges at higher intensities whereby rod signals enter neighboring cones via transmission through gap junctions . Signals from rod photoreceptor cells traverse the retina through at least three different pathways \u20133. The punctions \u201311. Therunctions , 13. Theunctions \u201320. Whilunctions . Other runctions and synaunctions . Howevervia the tertiary pathway involving direct contacts between rods and cone OFF bipolar cells. We expressed channelrhodopsin-2 (ChRh2) in rods and/or cones, allowing us to drive these two cell types independently with good temporal precision. Optogenetic activation of ChRh2 evokes depolarizing responses in rods or cones whereas activation of endogenous opsins evokes hyperpolarizing responses. Optogenetic activation therefore evokes inward currents in OFF cells at light onset rather than outward currents evoked by activation of endogenous opsins in rods and cones.In the present study, we combined optogenetic stimulation of rods and cones along with genetic elimination of exocytotic calcium sensors and gap junctions to distinguish the pathways carrying rod and cone signals to OFF \u03b1 ganglion cells. We were interested in evaluating the strength of synaptic inputs entering \u03b1 ganglion cells via direct synapses from cone OFF bipolar cells.\u03b1 RGCs are the most sensitive RGCs in mouse retina , 24. BotHRGP-Cre, Syt1flox (Syt1: MGI:99667), and Syt7flox mice have been described previously (Rho-iCre (RRID : IMSR_JAX:015850) mice were also obtained from Jackson Labs at room temperature for 15\u201330 min to aid in penetrating the inner limiting membrane . Retinas2/5% CO2. Except where noted, we blocked glycine receptors with strychnine (1 \u03bcM). During ganglion cell recording, GABA receptors were inhibited by supplementing Ames medium with picrotoxin (100 \u03bcM) or gabazine (10 \u03bcM). Early experiments used gabazine but later studies\u2013including pharmacological and gene knockout studies\u2013used picrotoxin. While picrotoxin is more effective than gabazine in blocking GABAc receptors . The use of BAPTA eliminated retrograde signaling effects (Whole cell recordings were obtained on an upright fixed-stage microscope (Olympus BX51) under a water-immersion objective (40x or 60x). Recording electrodes were fabricated from borosilicate glass pipettes to yield a tip resistance of 5\u20137 M\u03a9. Pipettes were filled with solution containing (in mM): 110 Cs gluconate, 8 NaCl, 1 CaCl effects .Recordings were performed in voltage clamp using an Axopatch 200B amplifier (Axon Instruments/Molecular Devices) and digitized with an ITC-18 interface (Heka Instruments). Data were acquired with AxoGraph X acquisition software and analyzed with Clampfit (Axon Instruments). Membrane currents were filtered at 5 kHz. RGCs were held at \u221260 mV. Voltages were not corrected for liquid junction potentials (Gluconate pipette solution: 12 mV).ChR2 was activated by a 1 ms pulse of 490 nm light from an LED . The voltage driving the LED was regulated by a computer-controlled analog input. For experiments reported here, we chose a voltage that consistently generated saturating responses (5 V).Confocal images were obtained using Nikon Elements software and a laser confocal scanhead (Perkin Elmer Ultraview LCI) equipped with a cooled CCD camera (Hamamatsu Orca ER) mounted on a Nikon E600FN microscope. Fluorescent excitation was delivered from an argon/krypton laser at 488, 568, or 648 nm wavelengths and emission was collected at 525, 607, and 700 nm, respectively. Filters were controlled using a Sutter Lambda 10\u20132 filter wheel and controller. The objective was controlled using a E662 z-axis controller (Physik Instrumente). Image contrast and brightness was adjusted using Nikon Elements and Adobe Photoshop software.Statistical analysis and data visualization were done using GraphPad Prism 9. Data were analyzed with paired and unpaired t-tests, as well as ordinary one-way ANOVA. We adjusted for multiple comparisons using the \u0160\u00edd\u00e1k method. The criterion for statistical significance was set at \u03b1 = 0.05. Data in the text and figures are reported as mean \u00b1 SD.To compare rod and cone inputs into ganglion cells, we used Ai32 mice that express a ChRh2/EYFP fusion protein in the presence of Cre-recombinase. To express ChRh2 in rods or cones, we crossed these mice with Rho-iCre or HRGP-Cre mice, respectively. We used blue LED light to activate ChRh2 in photoreceptor cells and recorded synaptic currents in RGCs using a flatmount retinal preparation . In the As illustrated in Optogenetic activation of ChRh2 depolarizes cones whereas activation of endogenous cone opsin hyperpolarizes cones. Optogenetic activation therefore evokes inward currents in OFF RGCs at light onset whereas activation of endogenous opsins evokes outward currents at light onset and inward currents at light offset EndogenoOptogenetic stimulation of ON RGCs \u2013C evokedvs. transient cells. Instead, OFF transient \u03b1 RGCs were distinguished from OFF sustained \u03b1 RGCs by the presence of prominent low-voltage-activated T-type Ca2+ currents in the former (We targeted OFF \u03b1 RGCs that receive rod and cone inputs for these experiments. In the presence of GABA antagonists (gabazine or picrotoxin) and the glycine receptor antagonist strychnine, responses of OFF cells to optogenetic stimulation of rods (n=8) and cones (n=12) reversed at positive potentials consistent with excitatory inputs. This is illustrated in e former , 38, 39.e former , 40, 41.+ channel blocker QX314 in the recording pipette. Contributions of Na+ currents produced an abrupt acceleration of the initial inward optogenetic current. As illustrated by example responses in + currents declined during the first few minutes of recording as QX314 entered the cell through the patch pipette. When reporting amplitude or latency of peak inward currents we did not include current components that showed evidence of sodium channel contributions.Excitatory synaptic currents evoked in OFF \u03b1 RGCs by optogenetic stimulation of cones or rods typically consisted of both fast and slow components. In some cells, the initial fast component was only an inflection during the rising phase of the inward current e.g., . We inclWe compared currents in OFF \u03b1 RGCs driven by optogenetic stimulation of rods or cones. Optogenetic stimulation of rods in Rho-iCre/Ai32 mice evoked responses in ganglion cells with similar waveforms as cone-driven currents in HRGP-Cre/Ai32 mice . Early eIn most of our experiments, the primary pathway from rod bipolar cells to OFF ganglion cells was blocked by using strychnine to inhibit glycinergic synapses between AII amacrine cells and OFF cone bipolar cells. To evaluate contributions of the primary pathway, we compared optogenetic stimulation of rods in Rho-iCre/Ai32 mice with and without strychnine. Strychnine (1 \u03bcM) reduced both amplitude . LatencyWe found no differences in paired pulse depression between rod and cone-driven pathways. In the presence of strychnine, the recovery from paired pulse depression was similar whether release was driven by rods in Rho-iCre/Ai32 mice or cones in HRGP-Cre/Ai32 mice . This isThe amplitude and latency to the initial peak current did not differ significantly between OFF-transient and OFF-sustained RGCs for either rod- or cone-driven cells , F. The The diagram at the left of When added in the presence of strychnine, L-AP4 (20 \u03bcM) caused no further decrease in current amplitude or charge transfer . Strychnvia gap junctions) or tertiary rod pathways (direct rod inputs to cone OFF bipolar cells).Consistent with other studies \u201347, we fTo examine contributions of rod-cone gap junctions to these currents, we tested effects of the gap junction blocker MFA (100 \u03bcM). Summary data are shown in In a second approach to testing gap junction contributions to rod-driven currents, we applied the D2/D4 dopamine receptor agonist quinpirole (1\u20133 \u03bcM). Quinpirole has been shown to uncouple gap junctions between rods and cones . Like MF2+ sensor used by cones to mediate synaptic release of glutamate-filled vesicles. Syt1 also controls a fast form of release from rods and eliminating Syt1 from rods and cones completely abolishes ERG b-waves to generate conditional Syt1 knockouts that lacked Syt1 in both rods and cones. Eliminating Syt1 from both rods and cones completely abolished responses to optogenetic stimulation of photoreceptors . OptogenWe combined pharmacology, knockouts, and optogenetics to analyze the pathways by which rods signals travel through the mouse retina. Blocking glycinergic synapses between AII amacrine and cone OFF bipolar cells with strychnine reduced optogenetic currents by ~1/3. This suggests that with optogenetic stimulation, ~1/3 of the rod input into the OFF pathway flows through inhibitory glycinergic synapses from AII amacrine cells to cone bipolar cells. This is a lower bound estimate since the activity of rod bipolar cells is nearly saturated by glutamate release in darkness and so aIn the presence of strychnine to block the primary rod pathway, large, fast currents could be evoked by optogenetic stimulation of rods in both transient and sustained OFF \u03b1 RGCs. This was true even after eliminating glutamate release from rods by genetic deletion of Syt1 and Syt7. Since eliminating glutamate release from rods should eliminate both primary and tertiary rod pathways, the large responses evoked by optogenetic stimulation of rods in the presence of strychnine must have arisen solely from the secondary pathway. Strong optogenetic stimuli may engage pathways that normally operate at higher light levels and so the secondary pathway may play a particularly prominent role in our experiments , 11.With the primary pathway blocked by strychnine, the latency of optogenetic responses evoked by stimulation of rods was 4.5 ms longer in OFF RGCs than responses evoked by optogenetic stimulation of cones. Cones have an average membrane capacitance of 6.2 pF and input resistance of 0.53 G\u03a9 suggestiIn the presence of strychnine to suppress the primary pathway, we inhibited the secondary pathway by inhibiting rod-cone gap junctions with MFA or by activating D2 receptors with quinpirole. We also tested Cx36 KO mice that lack gap junctions between rods and cones. Under these conditions, the tertiary rod pathway involving direct inputs from rods to cone OFF bipolar cells should be the major or sole remaining pathway. And under these conditions, optogenetic stimulation of rods generally evoked small, slow responses in OFF RGCs more consistent with poly-synaptic inputs than direct inputs from cone OFF bipolar cells. However, we saw fast responses in two RGCs from mice lacking Cx36 suggesting a subset receive direct contacts from OFF bipolar cells that in turn receive direct input from rods.c receptors that may not have been fully blocked by picrotoxin used in most of our experiments, including those with Cx36 KO mice (In mice lacking Cx36, the slow kinetics of responses evoked by optogenetic stimulation of rods suggested they arrive through a pathway that involves amacrine cells. One possible sign-conserving pathway from cone OFF bipolar cells would be from cone OFF bipolar cells \u2192glutamatergic monopolar amacrine cells \u2192 OFF RGCs . Another KO mice .Anatomical evidence suggests that OFF bipolar cells provide greater direct input into transient OFF \u03b1 cells than OFF sustained cells , 28. ConRecordings from individual rods showed that Syt7 contributes to synaptic release when stimulated with long depolarizing stimuli . EliminaHow do our results on rod pathways compare to previously published studies? Using multielectrode arrays to study mouse retina, Seilheimer et al. saw sevevia the tertiary pathway, but a subset of OFF transient cells may be specialized to carry this sort of information.Pasquale et al. found thIn summary, our data suggest that in addition to significant contributions from the primary rod pathway, much of the remaining input into OFF \u03b1 ganglion cells of the mouse retina involves transmission through gap junctions to cones. With optogenetic stimuli, the tertiary OFF pathway provides slow indirect input to most OFF RGCs but a subset of RGCs showed fast responses consistent with direct input from cone OFF bipolar cells. These fast direct connections may be particularly important for informing the brain about fast rod-mediated responses under mesopic conditions ."} +{"text": "THAP1 gene. She underwent bilateral posteroventral globus pallidus interna (GPi) deep brain stimulation (Medtronic Activa PC) implantation at the age of thirty-one years. She responded well to the deep brain stimulation even after more than 8 years post-surgery though she needs botulinum toxin injection for her cervical dystonia.A 21-year-old woman of south Asian origin presented with cervical dystonia which had progressed over the previous three years. Her symptoms started as writer\u2019s cramp since the age of seven years. She did not respond to medications and needed botulinum toxin injection for generalised dystonia. Subsequent whole genome sequencing revealed a likely pathogenic c.98G>A p.(Cys33Tyr) heterozygous variant in the THAP1 are the second most common cause of isolated monogenic dystonia after TOR1A [THAP1 with a previously reported variant now classed as pathogenic, with a very good response to globus pallidus interna (GPi) DBS after almost nine years of surgery demonstrated by video assessment.Genetic variants in er TOR1A . We repoA 21-year-old woman of south Asian origin presented with cervical dystonia which had progressed over the previous three years. Her symptoms started as writer\u2019s cramp since the age of seven years and had progressed with overflow movements. She was treated with tetrabenazine up to 25 mg three times daily and trihexyphenidyl up to 2 mg TDS but they were ineffective and caused side effects. Botulinum toxin (BoNT) type A injections for cervical and upper limb dystonia were initially effective, but the duration of benefit progressively declined despite abobotulinum toxin A dose of 250 units in the right sternomastoid and left splenius capitis each, 150 units in the left levator scapulae, and 100 units in the left trapezius muscle. She developed dystonic posturing of the left lower limb and laryngeal involvement in her mid-20s and was referred for deep brain stimulation (DBS) surgery at the age of 30.She underwent bilateral posteroventral globus pallidus interna (GPi) DBS (Medtronic Activa PC) implantation at the age of 31 heterozygous variant in the hat time .THAP1 was reported as a cause of monogenic isolated dystonia syndrome with a broad spectrum of genetic variants [DYT-THAP1 patients had been unsatisfactory or variable, and some patients may even deteriorate after an initial good response [45THAP1 mutation [TOR1A in which most patients harbour the same genetic variant. More recent reports suggest a more favourable response to GPi DBS in 14 patients with DYT-THAP1, in whom 11 were classed as responders, with 58% reduction in Burke-Fahn-Marsden motor score (BFM-M) after a median follow-up of 4 years and 10 months [THAP1 variant as we report here showed 58% reduction in BFM-M score after only one year of follow-up [THAP1 dystonia [The long-term motor response after pallidal DBS in DYT-response 456. The 0 months . The patollow-up . Krause dystonia .THAP1. One patient from India with a novel THAP1 frameshift deletion mutation also showed a positive response to GPi DBS even after ten years, with particular response of lower limb and truncal dystonia [THAP1 gene showed a good response to GPi DBS; however, this was during only two months of follow up, unlike our patient who had a sustained DBS response even after over eight years post-implantation [Other recent reports expand the spectrum of DBS response in DYT-THAP1 variant previously classed as a variant of uncertain significance, now classified as pathogenic, with a prolonged and sustained response to GPi DBS. Given the marked clinical and genetic heterogeneity of DYT-THAP1 and variability reported in DBS response, such reports add to the ability of clinicians to counsel prospective candidates for DBS in monogenic dystonia.We report a The additional files for this article can be found as follows:10.5334/tohm.774.s1Supplementary Table.Supplementary table: Cervical and generalised dystonia severity scales calculation before and after deep brain stimulation (DBS) surgery.10.5334/tohm.774.s2Supplementary figure(s).Supplementary figure: preoperative planning of leads in the right (2A) and left globus pallidus interna (2B) and the post operative imaging of the placements of the leads in the right (2C) and left globus pallidus interna(2D). The right globus pallidus internal lead is within 2 mm of preoperative planning site and the left lead was exactly at the planned location in the left globus pallidus."} +{"text": "Chat Generative Pre-training Transformer (ChatGPT) is a popular artificial intelligence-based chatbot that rapidly provides responses to online inquiries. As a language-processing \u2013 rather than a scientific \u2013 tool, ChatGPT does not guarantee accuracy. The United States Food and Drug Administration (FDA) labels medications with significant risk of serious or life-threatening adverse effects with boxed warnings (BW). We sought to assess the accuracy of ChatGPT regarding FDA BW for antibiotics.(1)\u2002\u201cMatching\u201d: ChatGPT accurately reported both existence and content of BW or its absence;(2)\u2002\u201cInaccurate\u201d: ChatGPT accurately reported a BW but erred on some aspects of the content; and(3)\u2002\u201cIncorrect\u201d: ChatGPT misidentified existence of BW, or listed incorrect adverse effects (AE)FDA labels for commonly-used antibiotics were categorized through the DailyMed database of the National Library of Medicine (Table 1). ChatGPT-3.5 was then queried regarding FDA BWs using the following query: \u201cAre there any boxed warnings on the FDA label for XX [medication] and if so, what are they?\u201d Responses were sorted into 3 categories:Of the 41 antibiotics queried, 9 have FDA BWs and ChatGPT correctly identified the existence of all 9. However, ChatGPT correctly matched the AE of the FDA BW in only 3 instances (33%); in five instances (56%), ChatGPT listed inaccurate AEs (either under- or overinclusive) and in one (11%), ChatGPT listed an incorrect AE. For the 32 antibiotics without an FDA BW, ChatGPT reported that 23 (72%) had a BW . Figure 2 provides some examples of incorrect ChatGPT claims regarding AEs.ChatGPT reported inaccurate and incorrect information about FDA BW for antibiotics. Clinicians should be aware of this new source of information for patients.All Authors: No reported disclosures"} +{"text": "Dear Editor,1 mHCC displays more complicated intratumor heterogeneity (ITH) and clonal evolution course which decreases the efficacy of clinical treatments.2 DNA damage repair (DDR) alterations have been put forward to be a prominent contributor to ITH.3 However, little is known about the role of DDR alterations in the genetic and epigenetic evolution of mHCC.Hepatocellular carcinoma (HCC) ranks the fourth most lethal cancer worldwide and over 50% of cases are diagnosed as multifocal HCC (mHCC) with dismal prognosis.TP53, AXIN1, KEAP1, and TTN are the most frequently mutated genes among the patients. The phylogenetic trees constructed by non-silent mutations of all patients showed branched evolutionary patterns on multiple lesions (MLs) including primary tumor, satellite nodule and portal vein tumor thrombus of seven mHCC patients . In line with genetic ITH, mHCC also exhibited a high level of methylation heterogeneity between lesions and between patients and oncogenes (KEAP1) were variously methylated at their CGI-promoters achieved partial response (PR) or stable disease (SD) while the other one (HCC5) achieved progressive disease (PD) (Supplementary Table +T lymphocyte densities and PD-L1 expression were significantly higher in responders (PR and SD), compared to non-responder (PD) Fig. . Our datIn summary, the present work provided a comprehensive evaluation of the genetic and epigenetic ITH of mHCC and a novel understanding of the importance of DDR alterations that might be implicated in tumor evolution. Of the patients receiving immunotherapy, DDR alterations in mHCCs may be a potential predictor for immunotherapeutic efficacy.Supplementary MaterialsData Set 1"} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-023-40935-7, published online 25 August 2023Correction to: The original version of this Article contained an error\u00a0in the Data availability section, where the SRA Bioproject accession number for DNA sequences was incorrect.\u201cThe datasets (fastq files) generated and/or analyzed during the current study are available in the NCBI Sequence Read Archive (SRA) repository under BioProject accession number PRJNA855317, and individual library accession numbers are listed in the Supplementary Data S1 file. Variant call format (vcf) files are available from the corresponding author upon request.\u201dnow reads:\"The datasets (fastq files) generated and/or analyzed during the current study are available in the NCBI Sequence Read Archive (SRA) repository under BioProject accession number PRJNA957102, and individual library accession numbers are listed in the Supplementary Data S1 file. Variant call format (vcf) files are available from the corresponding author upon request.\"The original Article has been corrected."} +{"text": "Cystic fibrosis (CF) is a genetic condition that causes abnormal mucus secretions in affected organs. MUC5AC and MUC5B are gel-forming mucins and frequent targets for investigations in CF tissues. Our objective was to qualify MUC5AC and MUC5B immunohistochemical techniques to provide a useful tool to identify, localize and interpret mucin expression in ferret tissues.MUC5AC\u2212/\u2212 and MUC5B\u2212/\u2212 ferrets. Qualified techniques for MUC5AC and MUC5B immunohistochemistry will be useful tools for mucin tissue studies in CF and other ferret models.MUC5AC and MUC5B mucins were detected most commonly in large airways and least in small airways, consistent with reported goblet cell density in airway surface epithelia. We evaluated whether staining method affected the detection of goblet cell mucins in serial sections of bronchial surface epithelia. Significant differences between stains were not observed suggesting common co-expression MUC5AC and MUC5B proteins in goblet cells of airway surface epithelia. Gallbladder and stomach tissues are reported to have differential mucin enrichment, so we tested these tissues in wildtype ferrets. Stomach tissues were enriched in MUC5AC and gallbladder tissues enriched in MUC5B, mucin enrichment similar to human tissues. Mucin immunostaining techniques were further qualified for specificity using lung tissue from recently generated Cystic fibrosis (CF) is a life-limiting condition caused by mutations in the CF transmembrane conductance regulator (CFTR) , 2. ClinMucins are high molecular weight glycoproteins that provide the characteristic viscoelastic features of mucus. In the respiratory tract, MUC5AC (goblet cells) and MUC5B are the major gel-forming mucins . As partThe aims of the current study were to qualify the MUC5AC and MUC5B immunostaining techniques through use of control ferret tissues including the lung.MUC5AC\u2212/\u2212 and MUC5B\u2212/\u2212 ferrets were obtained from an ongoing separate phenotypic study of these novel ferret models. Disruption of the two mucin genes was achieved using Cas9/gRNA ribonuclear protein complexes into ferret zygotes followed by adaptive transfer into pseudo-pregnant jills [Archival paraffin-embedded tissue blocks from wildtype (WT) ferrets or those with ongoing research projects at the University of Iowa. All studies were performed under the approval and guidance of the University of Iowa Animal Care and Use Committee and followed all pertinent federal/national/international standards of care. Adult WT ferrets were used to evaluate mucin expression in select tissues that are relevant to CF research. Additionally, lung tissue of recently developed nt jills . These eIn tissue sections (~\u20094\u00a0\u00b5m), diastase-pretreated periodic acid Schiff (dPAS) histochemical stain was applied to detect and localize mucus in tissues . BaselinRepresentative high-resolution digital images were collected and analyzed . Area of immunostaining relative to total area of airway surface epithelium produced the area fraction of immunostaining. These results were statistically analyzed with either two-way ANOVA or Kruskal\u2013Wallis test as warranted using Prism software . Airway epithelia height and muciPositive and negative cellular or tissue expression of protein targets are useful to qualify the specificity and utility to immunohistochemical techniques , 23. We Healthy gallbladder and stomach tissues have been reported to have distinct tissue enrichment of MUC5AC in stomach and MUC5B in gallbladder , 25. We MUC5AC\u2212/\u2212 ferret and a MUC5B\u2212/\u2212 ferret) that are being phenotypically characterized for an ongoing separate study. We evaluated serial sections of a bronchus from a MUC5AC\u2212/\u2212 ferret and both the surface epithelium and submucosal glands had MUC5B\u2009+\u2009immunostaining, but MUC5AC immunostaining was negative consistent with the tissue genotype can greatly influence immunostaining and digital image analysis , 41, 55."} +{"text": "Conserv Physiol 11(1): coac086; doi: https://10.1093/conphys/coac086In the originally published version of this manuscript, the figures within the paper are missing their respective captions and an incorrect supplementary file was uploaded.This error has been corrected."} +{"text": "Amazona viridigenalis ) primarily occupy vegetated urban rather than natural areas. We investigated the utility of raw vegetation indices and their derivatives as well as elevation in modelling the Red\u2013crowned parrot\u2019s general use, nest site, and roost site habitat distributions. A feature selection algorithm was employed to create and select an ensemble of fine\u2013scale, top\u2013ranked MaxEnt models from optimally\u2013sized, decorrelated subsets of four to seven of 199 potential variables. Variables were ranked post hoc by frequency of appearance and mean permutation importance in top\u2013ranked models. Our ensemble models accurately predicted the three distributions of interest . Maintaining existing preferred vegetation types and incorporating them into new developments should support the persistence of Red\u2013crowned Parrots in southern Texas.Texas Rio Grande Valley Red\u2013crowned Parrots (Psittaciformes: Amazona viridigenalis ) found in the southern United States of America (USA) are typically absent from natural areas but instead seem to prefer certain highly\u2013modified urban areas ) was declared extinct in 1939, and the Thick\u2013billed Parrot has been considered functionally extinct throughout its historical range within the USA since 1995 .\u201375.73\u201375rotocols . Eighty a et al. ) using Aa et al. .Presence data will not be made generally available to the public to protect the parrots from disturbance and poaching; however, the data can be requested via the eBird and iNatWe generated 10,000 background points, the MaxEnt default , separattopography (n = 1), (2) raw vegetation index (n = 8), (3) multitemporal vegetation index difference (n = 4), (4) percent cover of preferred ranges of raw vegetation index values (n = 24), (5) raw vegetation index textures (n = 48), (6) texture of binary preferred/non\u2013preferred ranges of raw vegetation index values (n = 48), and (7) percent cover of preferred ranges of vegetation index texture values , each of which falls into one of seven categories, in our SDM efforts: (1) Each predictor feature was a 9.5 m resolution raster projected to the 1983 North American Datum (NAD83) that covered the study area plus a one km buffer, which was added to eliminate edge effects introduced by the focal neighborhood calculations used to derive certain feature types. Feature rasters were converted from floating-point to integer format with a maximum of seven single digits to reduce raster size and computational complexity. Additionally, we applied a few additional non\u2013linear transformations to certain feature rasters to magnify differences between pixel values . All feae.g., pieced together like a puzzle) using ArcMap. Finally, we resampled the resulting raster to 9.5 m to create the elevation feature using information derived from spectral reflectance data ) they often prefer to nest in ). The tle value .i.e., novel information contributed by a given variable added to each of the 250 top\u2013performing models it appeared in over three replications) and total frequency of appearance in models , respectively. While both of these measures are important for evaluating variable performance, we gave more weight in the rankings to mean permutation importance as it is somewhat more relevant. We reported the ranking and correlation group number of each set of predictor variables (top 30) associated with their respective set of 250 top\u2013performing models. Variable correlation groupings were formed under top ranking variables to include other closely ranked variables correlated at |0.5| or higher by placing variables in multiple correlation groups where they have the closest ranking below the top ranked group member.We used the updated methodology of Tracy et al. to rank psa ) in Ecuador ) in Costa Rica ) presence was not random ) also prefers habitat with moderate, rather than high or low levels of vegetative cover ), which also occupy vegetatively heterogenous habitats in Venezuela ) woodlands within reasonable distances of each other).Variables derived using the larger focal window size (ibutions . This su>100 m2) . Occupyiion risk . As suchion risk . Since tion risk , we suspRed-crowned Parrots are highly mobile, strong fliers that movelevation. We suggest this indicates that the distributions of the various resource patches Red-crowned Parrots exploit across Rio Grande Valley are likely more important than the exact location of any single patch in isolation ) and Hispaniolan Parakeet as well as the Vulnerable Forest Red-tailed Black Cockatoo and the Critically Endangered Yellow\u2013crested Cockatoo are found in cities in the Dominican Republic, Western Australia, and Hong Kong, respectively ), Texas ebony (Ebenopsis ebano [Berland]), silver maple (Acer saccharinum), sycamore (Platanus spp.), and pecan (Carya illinoinensis [Wangenh]). A more expansive list of plant species consumed or used for nests by Red\u2013crowned Parrots can be found elsewhere ) distributions in the Rio Grande Valley of Texas using elevation and vegetation indices and their derivatives\u201d.This file contains the supplementary materials associated with the manuscript entitled \u201cModelling Red-crowned Parrot (Psittaciformes: (DOCX)Click here for additional data file.S1 Figelevation, was derived using parts of the 10 m United States Geological National Elevation Survey dataset.This variable, (TIF)Click here for additional data file.S2 Figbndvi08, was created by performing a simple calculation to the information contained in each of the three spectral bands of National Aerial Imagery Program (NAIP) images for 2008 , for 2008 .(TIF)Click here for additional data file.S3 Figdeltgrn10_08, was derived by subtracting raw 2008 Green Red Normalized Difference Index from raw 2010 GRNDI (grndi10).This variable, (TIF)Click here for additional data file.S4 Figi.e., 50%) of raw 2010 Normalized Difference Vegetation Index (NDVI) values while red dotted lines represent the central range of raw 2010 NDVI values of parrot presence points around the median raw 2010 NDVI value of 21.Blue dashed lines represent the central range ((TIF)Click here for additional data file.S5 Figbndvi_70_990, depicts the percent cover of a preferred range of raw 2008 Blue Normalized Difference Vegetation Index values . First, we calculated the preferred range of bndvi08 values Click here for additional data file.S6 Figbnd082c990, represents the 2nd-order Gray Level Co-Occurrence Matrix (GLCM) contrast texture of raw 2008 Blue Normalized Difference Vegetation Index values that was derived using a square 990 m focal window.This variable, (TIF)Click here for additional data file.S7 Figbnd08m70310, depicts the 2nd-order Grey Level Co-Occurrence Matrix (GLCM) mean texture of the binary preferred/nonpreferred range of raw 2008 Blue Normalized Difference Vegetation Index (BNDVI) values . Deriving this variable involved (1) calculating the central range of raw 2008 BNDVI values Click here for additional data file.S8 Figbnd082c99pc85, depicts the percent cover of preferred ranges of 2nd-order Grey Level Co-Occurrence Matrix (GLCM) contrast texture of raw 2008 Blue Normalized Difference Vegetation Index (BNDVI) values . Creating this variable involved (1) calculating the central range of 2nd-order GLCM contrast texture of raw 2008 BNDVI values derived using a square 990 m focal window, (2) creating a binary preferred/non-preferred raster from this range, and (3) calculating the percent cover of preferred areas from the binary raster using a square 990 m focal window.This variable, (TIF)Click here for additional data file.S9 FigThe jackknife of AUC for species graph (left) and response curve (right) results for each of the final models used to create the feature subset ensemble for the suitable general use habitat distribution.(TIF)Click here for additional data file.S10 FigThe jackknife of AUC for species graph (left) and response curve (right) results for each of the final models used to create the feature subset ensemble for the suitable nest site habitat distribution.(TIF)Click here for additional data file.S11 FigThe jackknife of AUC for species (left) and response curve (right) results for each of the final models used to create the feature subset ensemble for the suitable roost site habitat distribution.(TIF)Click here for additional data file.S1 Tables, standard deviation of the mean; Transf, transformation; Corr Group, correlation group; Imprt, importance; GLCM, Grey Level Co-occurrence Matrix; m, meters; b/w, between; diff, difference. For Corr Group: G, General Use; N, Nest Site; R, Roost Site. For Variable Rank, bolding indicates that the feature of interest appeared in (XLSX)Click here for additional data file.S2 TableIncludes the percentage of selected 12 top-selected models with at least one occurrence of variable type [% Models] and percentage among all 250 top-performing models with variable type being the top-performing variable by permutation importance (% Top). Percentage of variables used in top-selected 12 models does not include variables with 0.0% mean permutation importance .(XLSX)Click here for additional data file."} +{"text": "Ocimum sanctum L (Tulsi) has various properties like antibacterial, anti-inflammatory and anti-oxidant. To compare the effect ofthe local-drug delivery system containing 2% Ocimum sanctum L (Tulsi) as an adjunct to scaling and root planing (SRP).The main aimof the study was to evaluate the efficacy of Ocimum sanctum L (Tulsi) gel with Tetracycline fibers (Actisite) for the treatment ofperiodontitis patients. 40 subjects with periodontitis (pocket depth of 5 mm) were selected and divided into 2 groups Group I:Ocimum sanctum L (Tulsi) gel (n= 20) and Group II: Tetracycline fibers (Actisite) (n = 20). Clinical parameters assessed wereGingival Index , Plaque Index , Probing Depth and Clinical Attachment Loss were assessed at baseline, 1 month, 3 months, 6 months,8 months. Our results showed that Gingival index and Plaque index for for GROUP I: Ocimum sanctum L (Tulsi) and GROUP II:Tetracycline fibers (Actisite)are not statistically significant p>0.05 for baseline, at 1 month, 3 months, 6 months, 8 months.Probing depth and Clinical attachment are not significant p>0.05 for baseline, at 1 month, 3 months, 6 months, and statisticallysignificant difference seen at 8 months p<0.05. 2% Ocimum sanctum L (Tulsi) gel can be effectively used as an adjunct to scalingand root planing. When used as an adjunct to scaling and root planing, it helps in reduction of pocket depth and gain of clinicalattachment. Ocimum sanctum L (Tulsi) showed promising results when compared to Tetracycline fibers (Actisite). Periodontitis is a multifactorial disease that impairs the tooth's supporting structures. Chronic periodontitis, systemicdisease-associated periodontitis, and necrotizing periodontitis are examples of periodontitis. Periodontal disease is also caused bya local bacterial infection with pathogenic microflora in the periodontal pocket. The inflammatory process is triggered by microbialplaque and bacterial infection . BacteriAntibacterial drugs have been used in conjunction with mechanical debridement to treat periodontal infection. Because of thelimited access in the periodontal pocket, the efficacy of all methods is limited . Due toAntibacterial medications are used in conjunction with mechanical debridement to treat periodontal infections. The outcome islimited due to a lack of accessibility. Because the periodontal pocket provides an ideal environment for the growth of anaerobicpathogenic bacteria, antibiotics must reach the pocket's depth for effective treatment. The ideal requirements of the local drugdelivery should be delivered to the bottom of the pocket via the drug delivery system. It should only be effective againstperiodontal infections, not the commensal microbiome. The medication must be antibacterial. The planned dose must be sufficient tokill the target organism with no side effects. It should have a long shelf life. It must be biodegradable and biocompatible. Plantextracts are now commonly utilised as the primary component in mouthwash to reduce gingival inflammation, and gel versions arecommonly used to treat periodontitis .Ocimum sanctum L (Tulsi) belongs to the basil family Lamiaceae. Ocimum sanctum L (Tulsi) is an aromatic shrub. Ocimum sanctum L(Tulsi) was shown to have many qualities which can manage the interplay between the microbes and the body\u2019s immune response. Theinteraction between the host immune inflammatory mediators and pathogenic microorganisms denotes the development of theperiodontitis. Effectiveness of Ocimum sanctum L (Tulsi) in many formulations was studied. In situ or topical application of Ocimumsanctum L (Tulsi) in the form of liquid extract and in gel forms were researched Patients with generalized chronic gingivitis.[2] Patients in the age group of 20-65 years.[3] Systemically healthy subjects with Gingival index score, Plaque index score > 1 , Probing depth 5mm , Clinical attachmentloss 5mm at the time of examination.[1] Patients with gingivitis[2] Smokers[3] Antibiotic therapy within last 6 months of the study[4] Pregnant and lactating women[5] Patients undergone or having undergone periodontal therapy within the last 6 months of study.With the above Inclusion criteria and Exclusion criteria a total of 40 subjects were included in the study.40 subjects with chronic localized or generalized periodontitis with pocket depth of 5 mm. 40 periodontitis subjects were dividedinto 2 groups and sites were randomly selected to group I and group II by tossing a coin.Ocimum sanctum L (Tulsi) gel (n= 20)Group I: Group II: Tetracycline fibers (Actisite) (n = 20)Ocimum sanctum L (Tulsi) and Tetracycline fibers (Actisite) were given to subjects. Both groups received SRP and Ocimum sanctum L (Tulsi) gel (n= 20) was given to Group I and Tetracycline fibers (Actisite) was given to Group II. Theassessment criteria included Gingival Index (GI) score, Plaque Index (PI) score, Probing Depth (PD) and Attachment Loss (AL) scorethat were assessed at baseline, 1 month, 3 months, 6 months, and 8 months.Differences between the study groups were statistically analyzed by SPSS Software 23.0 version; PAIRED \"T\" TEST (Intra group) wasdone to analyse the difference between the groups. Inter group comparisons were analyzedby unpaired t test. Mean and Standard Deviation were assessed for statistical analysis and the results are tabulated. p< 0.05 wasconsidered a significant difference.Ocimum sanctum L (Tulsi) as LDD and 20 sites receivedTetracycline fibers (Actisite) as LDD. At the end of 1 month all sites were healed uneventfully and follow up was done up to 8months are discussed in tables (Ocimum sanctum L(Tulsi) treatment modalities were also observed.A total of 40 sites in 40 subjects were treated. 20 sites received n tables ,Table 7Plant extracts are possible sources of novel antimicrobial components especially against bacterial microorganisms. An importantfeature of plant extracts and their constituents is hydrophobicity which makes them divide the lipids portion of the cell membraneof bacteria and mitochondria interrupting the structures of cells and making them more absorbent. Plants have different forms ofbioactive compounds. It also has different forms of phytochemical compounds . TheantOcimum sanctum L (Tulsi) gel compared to Tetracycline fibers (Actisite)for the management of periodontal disease. Ocimum sanctum L (Tulsi) gel is effective demonstrating its potential use as efficientand in addition used as standard control for the management of periodontitis. Ocimum sanctum L (Tulsi) gel is used as mouthwashrinses and gel for the treatment of gingivitis and periodontitis. It is also used against oral microbes. Gupta et al. conducted atriple-blinded randomized controlled trial to test the efficacy of 4% w/v mouthrinse containing tulsi and 0.12% chlorhexidine. Itwas found that the mouthrinse containing O. sanctum was as effective in reducing gingivitis [Ocimum sanctum L (Tulsi) showed that it is effective in reducing gingival bleeding and gingivalinflammation. It also helps in reducing the Plaque. Ocimum sanctum L (Tulsi) showed no side effects when compared to Chlorhexidine(CHX) [Ocimum Sanctum (Tulsi) at 5% and 10%concentrations showed better inhibition zones against Aggregatibacter actinomycetemcomitans. They showed smaller inhibition zonesagainst Prevotella intermedia and Porphyromonas gingivalis. Hence proved that Ocimum Sanctum (Tulsi) can be utilized as an efficientadjunct and in addition to the regular periodontal treatment [et al.conducted a study about Ocimum Sanctum (Tulsi) gel that demonstrated possible anti-oxidant and anti-inflammatory effects. It is lesstoxic than brine shrimp nauplii. Ocimum Sanctum (Tulsi) proved to be the most favorable agent for the therapy of periodontalconditions [Ocimum sanctum L (Tulsi) (in gel form) as alocal drug-delivery system hence the study was planned to assess the efficacy of Ocimum sanctum L (Tulsi) gel compared toTetracycline fibers (Actisite) for the management of periodontal disease. However Adriana et al. observed similar reductions inmean plaque index gingival index sulcus bleeding index probing pocket depth; and gain in clinical attachment level using 1%chlorhexidine gel as an adjunct to SRP [Ocimum sanctum L (Tulsi) was usedas LDD in gel form we found that Our results showed that Gingival index and Plaque index for GROUP I: Ocimum sanctum L (Tulsi) andGROUP II: Tetracycline fibers (Actisite) are not statistically significant p>0.05 for baseline at 1 month 3 months 6 months8 months. Probing depth and Clinical attachment loss for GROUP I: Ocimum sanctum L (Tulsi) and GROUP II: Tetracycline fibers(Actisite) are not significant p<0.05 for baseline at 1 month 3 months 6 months and statistically significant difference seen at 8months p<0.05.The present study was conducted to assess the efficacy of ngivitis .Deepikane(CHX) . Mallikareatment . Ramamurnditions . There wt to SRP . In our Ocimum sanctum L (Tulsi) gel can be effectively used as an adjunct to scaling and root planing. When used as an adjunct toscaling and root planing, it helps in reduction of pocket depth and gain of clinical attachment. It is a beneficial antimicrobial,anti-inflammatory and anti-plaque agent. It is biologically well accepted by the oral tissues and showed good acceptability with noside effects by all the subjects in the study. Ocimum sanctum L (Tulsi) can be used as a local drug-delivery system for themanagement of periodontal disease. However, long term studies are required to validate the results.2%"} +{"text": "A new mental health service specialising in intellectual disabilities in Ireland was set up in January 2022. Its current compliment of staff includes is a Consultant Psychiatrist, Trainee Psychiatrist, Social Worker and Administrator. The current National Directive in Ireland is to prioritize Mental Health of Intellectual Disabilities services.The aim of the project is to establish the current baseline level of diagnostics and interventions within the new service. Our aim is to develop this service by implementing and following the gold standard guidelines and determine what extra resources does the service need.The first fifty case notes of patients assessed by the new service were inspected. The reviewer looked for evidence of the following clinical descriptions:Diagnosis of Intellectual Disabilities and its severity; Mental Capacity; Psychiatric Diagnoses; Physical health diagnoses; Medications and evidence of a Positive Behavioural Support Plan to manage complex challenging behaviours.The fifty patient audit contained 38 (76%) men and 12 (24% women) . One patient had Mild Intellectual Disabilities (ID), 39 (78%) had Moderate ID and 10 (20%) had Severe ID. All patents were very vulnerable and had limited or lacking Mental Capacity. Common diagnoses of the following were recorded in the following numbers and percentages; - Autism diagnosis 30 ( 60% ); Epilepsy 19 (38%); & Down Syndrome 9 (18%). A Formal Psychiatric diagnosis was identified in 26 (52%) of patients. Challenging Behaviour (severe and complex) was identified for 41 ( 82%) of the patients. The full breakdown of psychiatric diagnoses was \u2018Psychotic illness\u2019 \u2013 9 (18%); Anxiety \u2013 7(14%); Bipolar Affective Disorder 5 (10%): Depression \u2013 4(8%); Attention Deficit Hyperactivity Disorder (ADHD) 3 (6%); Obsessive Compulsive Disorder (OCD) \u2013 2(4%); Dementia \u2013 2(4%): Post Traumatic Stress Disorder (PTSD) \u2013 1 (2%); & Schizoaffective Disorder 1(2%). A Positive Behavioural Support plan (PBS) was available to support 33 (66%) of patients.42 (84%) of patients were prescribed antipsychotic medication. 12 (24%) were prescribed more than one antipsychotic. 20 (40%) were prescribed an antipsychotic without a formally documented diagnosis of a psychotic disorder. 12 (24%).The results of this first survey highlight areas in which the service can be improved. The service has requested funding for a Community Nurse and a Psychologist. Psychological evaluations and Positive Behavioural Support plans are essential for people with complex challenging behaviours. A Community Nurse should assist with Health Promotion and help supervise patients requiring Depot Antipsychotic medication or Clozapine. We also plan to set up a joint clinic with the Consultant Neurologist on a regular basis.None Declared"} +{"text": "Structural genes of mosaic BIB genomes are contributed by Butters while the immunity cassette is derived from Island3. Consequently, BIBs are morphologically identical to Butters (as shown by transmission electron microscopy) but are homoimmune with Island3. Recombinant phages overcome antiphage defense and silencing of the lytic cycle. We leverage this observation to propose a stratagem to generate novel phages for potential therapeutic use.Comparative analyses of mycobacteriophage genomes reveals extensive genetic diversity in genome organization and gene content, contributing to widespread mosaicism. We previously reported that the prophage of mycobacteriophage Butters (cluster N) provides defense against infection by Island3 (subcluster I1). To explore the anti-Island3 defense mechanism, we attempted to isolate Island3 defense escape mutants on a Butters lysogen, but only uncovered phages with recombinant genomes comprised of regions of Butters and Island3 arranged from left arm to right arm as Butters-Island3-Butters (BIBs). Recombination occurs within two distinct homologous regions that encompass In silico comparative analysis of mycobacteriophage genomes reveals extensive genetic diversity and widespread genome mosaicism. Mohammed et al., describe a novel mosaic recombinant phage named BIB (Butters-Island3-Butters) produced in situ by homologous recombination between the Mycobacteriophage Butters (cluster N) prophage within a lysogen and the invading Island3 (subcluster I1) genome. Mycobacteriophage Butters prophage-encoded factors block lytic infection of heterotypic Mycobacteriophage Island3 but the recombinant BIB genome contains genetic modules that allow escape from prophage-mediated defense and repressor-mediated lytic repression. The uptick in phage biology research coupled with a more accessible sequencing regime has led to sequencing thousands of phage genomes . ComparaThe prevalence of prophages in bacterial genomes likely iThe availability of large collections of sequenced mycobacteriophages provides opportunity to observe and investigate the relationship between phage genome mosaicism and exchange of antiphage defense systems. The HHMI-sponsored SEA-PHAGES program has led to the isolation of ca. 12,579 mycobacteriophages with over 2,000 genomes fully sequenced prophage and an infecting Island3 (subcluster I1) genome. These mosaic phages have been named BIBs to reflect the source of genetic segments that constitute their genome from left to right (Butters-Island3-Butters). Our data reveal a biological phenomenon where homologous recombination provides a means for the invading phage to overcome antiphage defense and the resident phage to escape from lytic repression. We discuss this phenomenon in the context of phage therapy as a measure where mixed phage lysates in coinfections are used to control bacterial infections.Mycobacterium smegmatis mc2155 as described in \u2265 1 \u00d7 109 PFU/ml), diluted with phage buffer , were used for immunity testing and PCR. Phamerator (https://defense-finder.mdmparis-lab.com/) was run on the webserver using the nucleic acid with CRISPR array detection (https://padloc.otago.ac.nz/padloc/myjobs/) was run at the website using the nucleotide fasta option on an LB agar plate (with 10\u2005\u00b5g/ml of cycloheximide [CHX] and 50\u2005\u00b5g/ml of carbenicillin [CB]). Phage lysates were serially diluted to 107 and spotted (3\u2005\u00b5l each) onto lawns of interest. Plates were incubated for 48\u2005h at 37\u00b0C. Phage growth was assessed at 24 and 48\u2005h. Plaque assays: Phage lysates were serially diluted. Ten microliters of each dilution was added to 250\u2005\u00b5l of the culture or lysogen to be infected and incubated for 10\u2005minutes at room temperature. Top agar (3.5\u2005ml) was added to the phage-bacteria mix and plated on LB agar plates (CHX/CB).Phage lysate was deposited onto 200 mesh copper grids coated with formvar and carbon (Ted Pella: 01810), rinsed with deionized water and stained with 2% ammonium molybdate. Samples were imaged on a JEOL 1200EX transmission electron microscope (TEM). Capsid and tail length measurements were done using ImageJ . MeasureSamples were derived from pure phage lysates or plaques picked into 100\u2005\u00b5l of phage buffer. Platinum\u2122 II Hot-Start Green PCR Master Mix (2X) (Invitrogen) was used. One microliter of each sample was used as template. Primers used are listed in Assuming 1 out of 10 phages within a plaque is non-BIB , then the probability of obtaining 1 non-BIB phage at 95% confidence interval is given by:n is the sample size of plaques to be screened to achieve a 95% confidence interval. From this equation we calculated that n must be at least 28.5 (rounded up to 30).30\u201338, and the antisense strands of genes 42\u201343 and gene 57 are expressed (2155(Butters)].Mycobacteriophage Butters is a temperate phage that encodes antiphage defense systems against several heterotypic phages . Analysixpressed . Analysexpressed . Furtherxpressed . While txpressed . To expl8 pfu of Island3. Nine of these putative Island3 DEMs were successfully sequenced.Single Island3 plaques, which typically constitute evidence of mutants, were not observed when serial dilutions of an Island3 high titer lysate were spotted on a Butters lysogen lawn . PromineSequencing of putative Island3 DEMs showed that genomes are recombinants with the left arm contributed by Butters, the middle segment derived from Island3, and the right arm contributed by Butters . These r29 or Island3 gene 32) (49 or Island3 gene 47), RecE, and RecT of about 84% and spans about 2,727\u2005bp of both genomes . This hogene 32) . By mappgene 32) . The secgene 32) . This seand RecT . Here, tand RecT . We estiand RecT , while tand RecT . In all and RecT , Table 1Morphological and immunity characteristics of recombinant BIBs were compared to their parental phages. Plaque morphologies of mycobacteriophages differ on different media ; plaque M. smegmatis mc2155, mc2155(Butters), and mc2155(Island3) show that BIBs efficiently infect mc2155(Butters) but are inhibited on an mc2155(Island3) lawn, due to Island3 immunity repressor activity were of mixed types that included wild type forms of Island3 and Butters and the reciprocal recombinant product IBI. Plaque assays were performed using Island3 to infect mc2155(Butters) to generate primary plaques. Primers were designed that would specifically PCR amplify each phage type across the recombinant regions . Plaque assays for 3 primary plaques were performed to generate secondary plaques on mc2155. In all cases, all thirty plaques were positive for a BIB-specific band but negative for all other phage types [Since 100% of plaques isolated and sequenced were recombinant BIBs, we investigated whether plaques produced from Island3 infection of mc regions . All priThe phenomenon of extensive mosaicism is well documented within phage genomes in general and within mycobacteriophages in particular . EvidencComparative genome analysis suggests that the observed was produced from homologous recombination events at two homologous regions between the Butters prophage and Island3 genomes. Over the entire genome, Butters and Island3 have 68% ANI. While there are several other homologous regions across the two genomes with ANI ranging from 43 to 80%, all sequenced BIBs recombined at the two regions with the highest ANI of 84 and 98%, respectively. It is unknown if recombinant phage form by homologous recombination at a single homologous region (to yield BI or IB) or by recombination at any other potential homologous regions, as no evidence of these possibilities was apparent.2155 for each biological replicate. BIBs plaques are morphologically similar to those of Island3, yielding bullseye plaques in both instances, but the defining concentric circles are markedly different in areas. Clearly, the exchanged genetic module derived from Island3 contributes genes that significantly affect plaque morphology phenotype. Yet, the inability to fully recapitulate the Island3 plaque morphology in BIBs suggests that other genes native to Butters portions of BIBs play a role in determining plaque morphology.Structural assembly genes determine phage morphotype . ExpecteBacillus subtilis prophage SP\u03b2 recombines with phi3T DNA during sporulation, but not in the absence of sporulation , 34 (repressor), 35 (HTH protein), and 36 (antirepressor) are annotated to function in lysis-lysogeny regulation. These genes are replaced by Butters genes 37 (integrase), 38 (repressor), and 39 (excise) which modulate lysis-lysogeny outcomes in Cluster N mycobacteriophages have homologs within the Butters exchange module corresponding to Butters genes 40, 45, 49, and 50 (RecE), respectively. Crucially, Island3 genes 38\u201340, 42\u201345, 46 (a type of HNH endonuclease) have no known homologs within the Butters module. If any of these Island3 genes play essential roles within the lytic cycle of Island3, then absence of a functional Butters homolog within the Butters module would also account for IBI nonviability. Finally, while recombination may be pervasive among groups of phages and lead to an increase in gene content variation, genome size remains conserved targeted by the Butters prophage-mediated defense system which will constrain its replication . Also, wiophages . Island3onserved , presumaE. coli was observed. Their findings supported the proposition that recombinants arise after several rounds of phage genome amplification Does recombination occur before Butters prophage induction or is the prophage induced before recombination? (2) Are there any Butters prophage- or Island3-specific factors that trigger the observed homologous recombination? (3) What is the source of recombineering enzymes that catalyze the observed recombination? Potential recombineering candidates include the host mcT system . (4) Canjkad210_Supplementary_DataClick here for additional data file."} +{"text": "As OA progresses, chondrocytes experience heightened catabolic activity, often accompanied by alterations in the extracellular environment\u2019s osmolarity and acidity. Nevertheless, the precise mechanism by which chondrocytes perceive and respond to acidic stress remains unknown. Recently, there has been growing interest in pH-sensing G protein-coupled receptors (GPCRs), such as GPR68, within musculoskeletal tissues. However, function of GPR68 in cartilage during OA progression remains unknown. This study aims to identify the role of GPR68 in regulation of catabolic gene expression utilizing an GPR68 and OA related catabolic genes was quantified using SYBR\u00ae green assays.We examined the expression of GPCR by analyzing high throughput RNA-Seq data in human cartilage isolated from healthy donors and OA patients. De-identified and discarded OA cartilage was obtained from joint arthroplasty and chondrocytes were prepared by enzymatic digestion. Chondrocytes were treated with GPR68 agonist, Ogerin and then stimulated IL1\u03b2 and RNA isolation was performed using Trizol method. Reverse transcription was done using the cDNA synthesis kit and the expression of GPR68 expression in cartilage associated with higher OA grades, underscoring a correlation between GPR68 expression and the severity of OA. Furthermore, IHC analysis of Gpr68 in murine cartilage subjected to surgically induced OA demonstrated elevated levels of GPR68 in knee cartilage and meniscus. Using IL1\u03b2 stimulated in vitro model of OA catabolism, our qPCR analysis unveiled a time-dependent increase in GPR68 expression in response to IL1\u03b2 stimulation, which correlates with the expression of matrix degrading proteases suggesting the role of GPR68 in chondrocytes catabolism and matrix degeneration. Using pharmacological activator of GPR68, our results further showed that GPR68 activation repressed the expression of MMPs in human chondrocytes.The transcriptome analysis revealed that pH sensing GPCR were expressed in human cartilage with a notable increase in the expression of GPR68 in OA cartilage which suggest a potential role for GPR68 in the pathogenesis of OA. Immunohistochemical (IHC) and qPCR analyses in human cartilage representing various stages of OA indicated a progressive increase in GPR68 was robustly expressed in human cartilage and mice and its expression correlates with matrix degeneration and severity of OA progression in human and surgical model. GPR68 activation in human chondrocytes further repressed the expression of MMPs under OA pathological condition. These results identify GPR68 as a possible therapeutic target in the regulation of matrix degradation during OA.Our results demonstrated that OA is cix in OA . The tarix in OA . IL1\u03b2 stogenesis . To datein vitro model that simulates catabolic processes characteristic of OA.During OA progression, chondrocytes undergo higher catabolism which often accompanied by fluctuations in extracellular osmolarity and acidity . These c2+ mobilization available from the NCBI-GEO database (http://biojupies.cloud), a web-based freely available application was used to analyze the differentially expressed genes (DEGs) to determine the expression of proton sensing receptors including GPR68 in healthy vs OA cartilage in the cartilage tissue during OA progression. The raw data were downloaded from healthy and OA cartilage tissues in human and mice (database . The humdatabase . BioJupiartilage . Raw datartilage . Multiplartilage . Mapped artilage . The mapGSE45793) . The GEOGSE63695) . All analyses were performed directly on the preprocessed data available online. We used the probe annotations from ChAMP . Genome om ChAMP to assigPrior to the initiation of the studies the study protocol was reviewed and approved by the Institutional Review Board (IRB) of Emory University, Atlanta, GA as a \u201cnot research involving human subjects\u201d as defined by DHHS and FDA regulations and that no informed consent was needed. All the methods used in this study were carried out in accordance with the approved guidelines and all experimental protocols were approved by the IRB of Emory University, Atlanta, GA (#IRB ID: STUDY00003990). Discarded and de-identified knee joint cartilage samples were collected from the patients undergoing joint arthroplasty at the Emory University Orthopaedics and Spine Hospital (EUOSH), Atlanta, GA.in vitro culture was maintained by plating the chondrocytes at high cell density (1 \u00d7 106/well of 6-well plate) and phenotype was verified by analyzing the mRNA expression of chondrocytes phenotype markers such as COL2A1, ACAN, and COL10A1 using real time PCR analysis.Macroscopic cartilage degeneration was determined by India ink staining and cartilage degeneration were graded according to Mankin scoring system . In this6/ well) in Dulbecco\u2019s modified Eagle\u2019s medium/Nutrient Mixture F-12 (DMEM/F-12) supplemented with 10% fetal calf serum (FCS), penicillin (100 units/ml), and streptomycin (100 mg/ml) for 2\u20133 days. When cells reached 80% confluency, OA chondrocytes were serum starved overnight and then stimulated with IL1\u03b2 (1 ng/ml) for 1, 3, 6, 12 and 24 h at 37 \u00b0C, in CO2 incubator under 5% CO2 and 95% air. Unstimulated chondrocytes were used as control.Primary human OA chondrocytes were seeded at high confluency in six well plate (1 \u00d7 10Human OA chondrocytes cultured in 6-well plate were treated with different concentration of Ogerin (1\u201310 \u00b5M) for 2 h followed by stimulation with IL1\u03b2 (1 ng/ml) in incomplete DMEM/F-12 medium without presence of serum. Ogerin (OGR) is a positive allosteric modulator (PAM) of GPR68 and known to activate GPR68 at physiological pH. Chondrocytes which were treated with 0.1% DMSO served as control for this experiment.GPR68, MMP13, and MMP3 was quantified using SYBR Green expression assay as previously described for 72 h were fixed in 10% neutral buffer formalin for 24 h, decalcified in 10% EDTA (pH 7.4) for 72 h, dehydrated with series of alcohols and then embedded in paraffin wax, sectioned at 5 \u03bcm thickness were stained with Safranin-O dye as described previously . The car2 asphyxiation, followed by cervical dislocation as secondary physical method of euthanasia.Male C57BL/6 mice were obtained from Jackson Laboratory . Twenty mice were randomly divided into two groups (Sham and DMM) of ten animals per experimental group as described previously and eachExperimental OA was induced by performing surgical destabilization of the medial meniscus (DMM) in the left knee of 12 weeks old male mice as described previously . BrieflyFull thickness human cartilage pieces resected from the human cartilage or intact knee joints harvested 12-week post-surgery from sham and DMM operated mice were fixed in 4% paraformaldehyde for 48 h. Samples were decalcified using 10% EDTA for 2 weeks and dehydrated cartilage pieces were embedded in paraffin and 5 \u03bcM thick sections were prepared. Deparaffinization of cartilage sections was performed by incubating histological section with xylene three time (5 min each) and then sections were rehydrated using various grade of alcohol in a decreasing concentration . The sections were washed with 1X TBS for 10 min and antigen retrieval was performed by incubating histological sections in 10 mM citrate buffer (pH 6.0) at 65 \u00b0C for 1 h using water bath. To analyze the immunoreactivity of GPR68, the endogenous peroxidase activity was neutralized by 3% hydrogen peroxide in 1X TBS, sections were blocked in 10% normal goat serum for 30 min at room temperature, and incubated in rabbit polyclonal anti-GPR68 antibody overnight at 4 \u00b0C. We also used recombinant rabbit IgG monoclonal antibody as negative isotype control. Sections were washed with 1X TBS and incubated with HRP-conjugated secondary anti-rabbit antibody for 2 h followed by washing with 1X TBS, and developed using a SignalStain\u00ae DAB substrate kit and images were captured BioTek Lionheart LX Automated Microscope as described previously , 2020a.post hoc test. The significance was set at P < 0.05.The data presented here represent the value of Mean \u00b1 SD. The statistical difference between control and experimental groups was performed by GraphPad PRISM using one-way ANOVA with Tukey\u2019s GSE106292) were determined in human cartilage across four different stages of human ontogeny including (1) embryonic cartilage isolated from limb bud condensations at embryonic 5-6 Week Post Conception (WPC), (2) presumptive cartilage derived from 17 WPC, (3) healthy juvenile cartilage, and (4) adult articular chondrocytes. In addition to human in vivo chondrogenesis, we also analyzed two stages of in vitro pluripotent stem cell (PSC) differentiation into chondrocytes at 14 and 60 days. Our analysis demonstrates that all there four members of proton-sensing orphan GPCR genes expressed in human cartilage, and GPR68 expression was highest in all the human cartilages analyzed (GSE110051) generated in immature and mature chondrocytes isolated from the mouse embryo and adult histological section using laser capture microdissection. Our analysis demonstrates variable degree of expression levels of all four GPCRs, with Gpr68 showing highest expression level in both immature and mature cartilage . After sanalyzed . We nextartilage . These rGPR68, we analyzed RNA-seq data performed on performed on 38 individuals including 18 healthy and 20 OA human knee cartilage tissues (GSE114007) compared to healthy cartilage (N = 18) (# P < 0.01) . Our res < 0.01) . These rGPR68 in OA pathogenesis, we analyzed its expression in human cartilage isolated from different stages of OA. High-grade OA cartilage showed severe cartilage degeneration as compared to low-grade OA cartilage (Mankin score \u2264 3) and exhibited loss of proteoglycan content as analyzed by Safranin-O and Fast green staining indicating the loss of cartilage ECM during OA progression (GPR68 was significantly higher in cartilage with high-grade OA (N = 10) compared to the cartilage with low-grade OA (N = 10) (P < 0.01). The increased expression of the GPR68 positively correlated with the severity of OA further reinforcing our hypothesis that GPR68 plays a unique role in the pathogenesis of OA.To further confirm the role of gression . Our IHCGpr68 in OA pathogenesis, we examined the expression level of GPR68 in murine cartilage of surgically induced OA. DMM surgery, which is transection of medial meniscotibial ligament, is a gold standard approach to model human post-traumatic OA (PTOA) (Gpr68 expression in mice cartilage of DMM joints as compared to sham operated knee joints (GSE45793) (N = 4/group) (P < 0.01). Collectively, data support our hypothesis that Gpr68 play a significant role in cartilage erosion during OA progression in mice.To further confirm of role of A (PTOA) by inducA (PTOA) . It has A (PTOA) , which aA (PTOA) . InteresA (PTOA) . These rSE45793) . region of GPR68 (P < 0.01). These results demonstrating hypomethylation of GPR68 promoter in severe OA cartilage are in concordance with increased GPR68 expression in high grade OA cartilage as shown in GPR68 in OA progression.DNA methylation plays a major regulatory role in gene transcription mainly tion way . The hyption way . To examSE63695) . We analof GPR68 . InteresGPR68 expression robustly increased in high grade human OA cartilage and DMM operated knee joints in mice as shown in GPR68 expression correlates with the degeneration of extracellular matrix. For this, we analyzed cartilage ECM content by Safranin-O staining which stain the sulfated GAG and proteoglycan levels of cartilage matrix in human cartilage explants and murine knee joint sections. Our analysis of human cartilage as shown in Since COL2A1, ACAN etc., for different time points. Our qPCR analysis showed that IL1\u03b2 treatment induced the expression of GPR68 in a time dependent manner in human OA chondrocytes (P < 0.01). Consistent with the increased mRNA expression levels, immunostaining of histological sections of human cartilage revealed that the GPR68 protein level was also markedly high upon stimulation with IL1\u03b2 . These rGPR68 and proteases mRNA expression in human OA chondrocytes. Furthermore, OA chondrocytes that express high level of GPR68 under stimulation with IL1\u03b2 showed positive correlation with expression level of MMP13 (Our analysis demonstrates that IL1\u03b2 is potent inducer of GPR68 expression in human OA chondrocytes and cartilage . We nextof MMP13 and otheof MMP13 . AltogetOur correlation studies as shown in GPR68 particularly in the damage and fibrillated areas of the human cartilage tissue from OA patients. Additionally, our results showed significant upregulation of Gpr68 expression, specifically in degenerated cartilage and inflamed meniscus of murine joint tissue during surgically induced OA model. Our data showed that GPR68 expression significantly increased in cartilage tissue isolated from OA patients with high grade disease severity showing a severe degeneration of extracellular cartilage matrix, when compared to cartilage from healthy volunteers or low-grade OA patients. Chondrocytes are the resident cell type present in cartilage. Our results showed that increased GPR68 expression in primary human chondrocytes stimulated with inflammatory cytokine IL1\u03b2 which induced OA like pathogenic situation. Using several correlation analyses, we showed that GPR68 expression positively correlated with degree of OA severity, cartilage matrix degeneration and expression of MMP13 and other matrix degrading proteases in human chondrocytes. Consequently, we hypothesize that GPR68 activity leads to an increased catabolic gene expression which triggers the activation of signaling cascades involved in further ECM degeneration and degradation of cartilage tissue leading to OA progression. Using selective activation of GPR68 by the treatment with small molecule agonist, the Ogerin, our results further showed that increase GPR68 activity inhibits the expression of matrix degrading proteases under OA like inflammatory conditions in human chondrocytes in vitro.GPR68 is a proton sensing orphan receptor which is activated upon binding to extracellular protons during acidosis or acidic stress. These pH sensors sense extracellular protons and stimulate different intercellular signaling pathways . Recent in vitro model of cartilage degeneration using human OA cartilage explants and in vivo model of surgically induced OA in mice to demonstrate the role of GPR68 in the degradation of extracellular matrix in OA pathogenic condition. Our results elucidated the positive correlation of GPR68 expression with loss of proteoglycan content in human OA cartilage and surgically induced OA in mice indicating the plausible role of GPR68 in OA pathogenesis.Osteoarthritis is the most common form of arthritis among older adults. OA is a degenerative joint disorder characterized by the breakdown of cartilage, leading to joint pain, stiffness, and loss of function. It is also one of the most common causes of physical disability among adults . InflammGPR68 mRNA and induced GPR68 expression demonstrated the repression of mRNA expression of several matrix metalloproteinases such as MMP13, and MMP3. Together, these results demonstrate a new role for a proton sensing orphan gene in the regulation of MMPs expression in chondrocytes during disease progression.During OA progression, cartilage tissue is damaged due to mechanical injury, trauma, aging, and inflammation. Degeneration of ECM is hallmark of OA pathogenesis . Extracein vitro studies including ours showed that IL1\u03b2 stimulation of articular chondrocytes leads to expression, and secretion of active MMP13 which cleaves the major components of ECM including type two collagen and aggrecan (in vitro model of OA, our results demonstrate the protective role of Ogerin and showed that GPR68 activation exhibited anti-catabolic effect by repressing the expression of matrix degrading proteases including MMP3 and MMP13. Human chondrocytes used in our study were derived from OA cartilage where significant matrix degeneration has already occurred, the ability of Ogerin to repress matrix degrading protease gene expression has significant implications to halt or slow down the cartilage erosion and thus halt the disease progression.Activation of inflammatory signaling has been shown to induce matrix degeneration in several degenerative disorders including OA . Severalaggrecan . Our resaggrecan . Using oin vitro and in vivo GPR68-/-mice are needed to determine the specific role of GPR68 in the transcription regulation of matrix degrading proteases genes during OA progression. We envision a multitude of follow-up studies using overexpression and knockout of GPR68 mice model aimed at thoroughly examining the role of GPR68 during OA progression in aging and post traumatic surgery model which will define the precise role of GPR68 in controlling the cartilage degeneration. Our future studies on the expression of GPR68 during aging and post-traumatic OA are likely to shed new light on the role of GPR68 in regulation of catabolic gene expression and pathogenesis of OA.Although data present here provide a clear role of GPR68 in the regulation of matrix degradation during OA progression. However, an in-depth mechanistic analysis of using genetic manipulation of GPR68 using in vitro, it is important to understand the phenotypes of in vivo GPR68 activation or GPR68 loss of function to accurately interpret its role in matrix degeneration. Additionally, our in vitro culture condition used physiological pH, owing to fact that GPR68 activity is higher under acidic condition, further studies are needed to determine the effect of pH on the activation of GPR68 in chondrocytes and its role in the regulation of ECM degeneration in cartilage explants. Future work investigating the effect of Ogerin in joint instability model of rat and mice in vivo will further expand our understanding of GPR68 in pathological states which will open a new avenue for potential of Ogerin as therapeutics in OA.Although this study provides first demonstration of role of proton sensing GPR68 in OA, there are some limitations of this study. Our studies are mainly correlative in nature and focusses on human chondrocytes GPR68 is expressed in human and mice cartilage. Our results demonstrated that GPR68 expression profoundly increased in cartilage and meniscus during surgically induced OA in mice. GPR68 expression in human and mice cartilage correlates with matrix degeneration and severity of OA progression. Additionally, stimulation of chondrocytes with inflammation results in induced expression of GPR68 and increased GPR68 activation further repressed the expression of matrix degrading proteases including MMP13 and MMP3 in OA chondrocytes indicating the plausible role of GPR68 in regulation of metalloproteinase gene expression. These results identify GPR68 as a possible therapeutic target in the regulation of matrix degradation during OA.In conclusion, the present study is the first to demonstrate that a proton sensing orphan gene 10.7717/peerj.16553/supp-1Supplemental Information 1Click here for additional data file.10.7717/peerj.16553/supp-2Supplemental Information 2Click here for additional data file."} +{"text": "Manic and depressive mood states in bipolar disorder (BD) may emerge from the non-linear relations between constantly changing mood symptoms exhibited as a complex dynamic system. Dynamic Time Warp (DTW) is an algorithm that may capture symptom interactions from panel data with sparse observations over time.The current study is the first to analyze a time series of depression and manic symptoms using DTW analyses in patients with BD. We studied interactions and relative changes in symptom severity within and between participants.The Young Mania Rating Scale and Quick Inventory of Depressive Symptomatology were repeatedly assessed in 141 patients with BD, with on average 5.5 assessments per patient every 3 to 6 months. DTW calculated the distance between each of the 27*27 pairs of standardized symptom scores. The changing profile of standardized symptom scores of BD patients was analyzed in individual patients, yielding symptom dimensions in aggregated group-level analyses. Using an asymmetric time-window, symptom changes that preceded other symptom changes yielded a directed network.The mean age of the patients was 40.1 (SD 13.5) years old, and 60% were female. Idiographic symptom networks were highly variable between patients. Yet, nomothetic analyses showed five symptom dimensions: core (hypo)mania (6 items), dysphoric mania (5 items), lethargy (7 items), somatic/suicidality (6 items), and sleep (3 items). Symptoms of the \u2018Lethargy\u2019 dimension showed the highest out-strength, and its changes preceded those of \u2018somatic/suicidality\u2019, while changes in \u2018core (hypo)mania\u2019 preceded those of \u2018dysphoric mania\u2019.Image:Image 2:Image 3:DTW may help to capture meaningful BD symptom interactions from panel data with sparse observations. It may increase insight into the temporal dynamics of symptoms, as those with high out-strength (rather than high in-strength) could be promising targets for intervention.None Declared"} +{"text": "Pseudomonas aeruginosa (Pa). It is widely accepted that increasing antibiotic resistance develops in Pa following repeated antibiotic exposure. With the advent of elexacaftor/tezacaftor/ivacaftor (ETI) modulator therapy, we wished to assess Pa antibiotic resistance to beta-lactam and fluoroquinolone (FQ) antibiotics in both children receiving ETI, and those not eligible for ETI (nonETI) who continue to experience recurrent pulmonary exacerbation (PEx) and require repeated antibiotic treatment.CF is a genetic disease characterized by chronic lung infection, often with We examined current patterns of antibiotic resistance over 3 years for a cohort of 11 children with CF having 5 or more positive sputum cultures for Pa between January 2020-December 2022 obtained from CF clinic and hospital PEx sputum samples. Chart review included: demographic data; dates of PEx/hospitalization; dates of CF clinic visits; dates and results of CF sputum cultures; and antibiotics used and dates for intravenous (IV) and oral (PO) treatment courses.Six males and 5 females were reviewed (6/11 [54%] were Hispanic), with 3/11 (27%) on ETI therapy (average age 14.6 yr), and 8/11 (73%) not on ETI therapy (average age 11.6 yr). Average age at acquisition of Pa was 10.2 yrs. The average number of cultures yielding Pa over 3 years was 11 for both ETI and nonETI subjects. Average number of IV antibiotic courses was 6 for nonETI, but 0 for ETI subjects, while the average number of oral (PO) FQ courses with ciprofloxacin/levofloxacin was 6 in nonETI, and only 1 in ETI subjects. No increase in antibiotic resistance was noted against any IV antibiotic over 3 years of observation (Table), but of the 9 subjects (both ETI and nonETI) who received PO FQ, 5 (55%) were documented to have at least one nonsusceptible isolate, as shown for one 16 year old nonETI subject .Current antibiotic/pulmonary management of children with CF was not associated with an increased risk of IV antibiotic resistance over 3 years of observation despite repeated exposure for PEx (mean of 6 IV antibiotic courses per nonETI subject), but susceptibility to FQ decreased in 55% of those treated over 3 years following repeated FQ oral therapy courses.All Authors: No reported disclosures"} +{"text": "During the COVID 19 pandemic children and young people (CYP) faced significant restrictions. The virus and restrictions also affected how health services could function, including mental health. Research analysing the COVID 19 pandemic is important to ensure dynamic and resilient service design in case of future national emergencies.To investigate the impact of COVID-19 lockdowns on CYP psychiatric admission trends during lockdowns 1 (started 26 March 2020) and 2 (started 20 November 2020) of the COVID 19 pandemic in England.Routinely collected, retrospective, English, administrative data looking at the CYP hospital admissions, length of stay and patient demographics were analysed using an interrupted time series analysis to compare pre-pandemic service use with service use during COVID 19 lockdowns 1 and 2. The analysis used an ordinary least squares (OLS) approach with Newey\u2013West standard errors to handle autocorrelation and heteroscedasticity.Image:Image 2:Image 3:During lockdown 1 and 2, psychiatric admissions for CYP were fewer and shorter. The rise in admissions for more deprived CYP and looked after children suggests these CYP may have been disproportionately affected by the pandemic.None Declared"} +{"text": "Escherichia coli (UPEC) that poses a serious challenge in the management of urinary tract infections (UTIs). The WHO has described antibiotic resistance in uropathogens as a key pressure point in the burgeoning global antimicrobial resistance crisis. Given this grim situation, we are exploring phage-encoded lysins as plausible alternatives.Atrocious use of antibiotics has led to the emergence of MDR in uropathogenic in silico strategy for the discovery and characterization of lysin sequences (seq) targeting E. coli cell wall and evaluating the bactericidal activity of these recombinant lysins using in-vitro assays.Our study involved an E. coli prophages in the database (using PHASTER). Lysozyme-like domain was observed in 9 out of 16 lysins. Their characterization depicted modular or globular structure. Based on the physicochemical properties, 7 out of 16 lysins were selected for cloning, expression, and purification as recombinant proteins for evaluating the bactericidal activity.Novel lysin sequences were searched by BLAST homology and by screening in vitro assays. Using static biofilm assay, lysin seq 5 (180 \u03bcg) showed efficient reduction (>50%) in the biofilm formed by ATCC UPEC 700928 strain. As per turbidity reduction method, lysin seq 5 (50 \u03bcg) showed 37% drop in OD600nm on UPEC 700928 strain after 3 h of incubation at 37\u00b0C. Using spot on lawn assay, lysin seq 5 (20 \u03bcg) exhibited lytic activity (zone of inhibition) on five drug-resistant clinical UTI isolates, which were pretreated with an outer membrane permeabilizer (OMP), viz., EDTA (0.3 mM).Among the several lysins screened, lysin seq 5 demonstrated the highest activity using Lysin seq 5 exhibited antibiofilm activity against UPEC 700928 strain as well as lytic activity against drug-resistant clinical UTI isolates. Screening of additional drug-resistant clinical isolates from UTI patients is underway."} +{"text": "The fluid status and causes of respiratory failure in patients with Ebola disease can be challenging to determine due to concomitant severe diarrhea, vascular leakage, and multiorgan failure. Point-of-care ultrasound (POCUS) could guide fluid resuscitation for patients in resource-limited settings. We present the clinical application of POCUS in the 2022 Sudan Virus disease Ebola outbreak in Uganda.Figure.Ultrasound findings from patient 2. 2A: Discrete B lines observed with an irregular pleural line; 2B: Inferior vena cava collapsibility <50% suggestive of adequate fluid resuscitation; 2C: parasternal short cardiac view with small pericardial effusion identified; 2D: apical cardiac view with small pericardial effusion redemonstrated.A clinical research team in Fort Portal, Uganda was trained in advanced Infection Prevention and Control (IPC) to care for patients with Ebola disease and in POCUS for septic shock. The clinical team learned a standardized lung and cardiac critical care ultrasound approach to evaluate patients in septic shock. The scanning procedure was used during the care of 6 patients with confirmed Sudan virus disease within 2 weeks of outbreak declaration to make critical patient care decisions like fluid management. Chest X-ray services and mechanical ventilation were not accessible. A handheld Philips Lumify device was kept in the biocontainment area and was used in full personal protective equipment. Scanning procedures included 12-zone lung ultrasound and 4-view cardiac ultrasound scans to determine causes of respiratory disease and evaluate fluid status.and another with a >50% IVC collapsibility (1.3 cm diameter). These findings contributed to a decision to continue fluid resuscitation.Among 6 patients , 5/6 were in their third week of illness, and 1/6 was in his second week of illness. Lung ultrasound abnormalities included discrete B-lines (5/6 patients), confluent B-lines (1/6), irregular pleural lines (3/6 patients), and subpleural consolidations (1/6 patients). Two participants in respiratory distress had cardiac POCUS. Both had hyperdynamic left ventricular function and one had a small pericardial effusion . One patient had inferior vena cava (IVC) collapsibility < 50% (1.5 cm diameter), Portable handheld ultrasound devices identified previously undescribed ultrasound features of Sudan virus disease. POCUS helped determine causes of respiratory failure and guided fluid resuscitation in a biocontainment ward without access to mechanical ventilation.All Authors: No reported disclosures"} +{"text": "Treatment resistant depression (TRD) is common with substance use disorder (SUD) and few studies demonstrated the effectiveness of medication-psychotherapy treatments in this populationTo compare the effeictiveness of ECT in the treatment resistant depression patients vs TRD with SUD patients.14 TRD patients with 14 TRD-SUD patients compared in terms of ECT treatment response rates at baseline, thhree months and six months of the follow up periond. Pateints completed Hamilton anxiety, Hamilton depression-21 items, Barrat 11 impulsivity and visual analog scales each follow up visit.+4.1 months following the ECT procedure. Patients received average 11.7+2.6 bilateral ECT treatments per series. Both groups responded well to ECT treatment in terms of response rates and side effects however there were higher rates of relapse at intermediate to long term follow up period at TRD-SUD group.Both groups completed ECT treatment between 2011-2018 with follow up of 12.3ECT seems to be an effective treatment for patiens of TRD-SUD. Moreover the response rates are equal to treatment reisstant depression cases without substance use disorder.None Declared"} +{"text": "Hidradenitis Suppurativa is a chronic inflammatory disease of which the pathogenesis is incompletely understood. Dermal fibroblasts have been previously identified as a major source of inflammatory cytokines, however information pertaining to the characteristics of subpopulations of fibroblasts in HS remains unexplored. Using in silico-deconvolution of whole-tissue RNAseq, Nanostring gene expression panels and confirmatory immunohistochemistry we identified fibroblast subpopulations in HS tissue and their relationship to disease severity and lesion morphology. Gene signatures of SFRP2+ fibroblast subsets were increased in lesional tissue, with gene signatures of SFRP1+ fibroblast subsets decreased. SFRP2+ and CXCL12+ fibroblast numbers, measured by IHC, were increased in HS tissue, with greater numbers associated with epithelialized tunnels and Hurley Stage 3 disease. Pro-inflammatory CXCL12+ fibroblasts were also increased, with reductions in SFRP1+ fibroblasts compared to healthy controls. Evidence of Epithelial Mesenchymal Transition was seen via altered gene expression of SNAI2 and altered protein expression of ZEB1, TWIST1, Snail/Slug, E-Cadherin and N-Cadherin in HS lesional tissue. The greatest dysregulation of EMT associated proteins was seen in biopsies containing epithelialized tunnels. The use of the oral Spleen tyrosine Kinase inhibitor Fostamatinib significantly reduced expression of genes associated with chronic inflammation, fibroblast proliferation and migration suggesting a potential role for targeting fibroblast activity in HS. Hidradenitis Suppurativa (HS) is a chronic inflammatory skin disease with an estimated prevalence of 1% globally . The molDermal fibroblasts are a heterogeneous population, with functionally distinct fibroblast subpopulations have been identified in normal skin , 12. MorThe therapeutic modulation of fibroblasts is an emerging area of research in fibroinflammatory diseases with existing therapies including Janus Kinase (JAK) inhibitors and Spleen Tyrosine Kinase (SYK) inhibitors demonstrating evidence of clinically relevant fibroblast modulation . These dIn this study we aimed to examine the presence of fibroblast subpopulations in lesional tissue (Frew 2019b) of HS coTGFB1, OSM, IL6, MMP1 and MMP3. These genes discretely clustered in lesional versus non lesional tissue including CDH1, CDH2, SNAI2, TWIST1, ZEB1 were not significantly differentially expressed between lesional and non-lesional HS tissue S2 Fig)S2 Fig). Transcriptomic findings illustrating differential gene expression of fibroblast sub-populations was validated using immunohistochemistry with primary antibodies against SFRP1, SFRP2 and CXCL12 proteins identified increased staining of TWIST1 , ZEB1 F, Snail/SDifferences in intensity of N-Cadherin staining were observed between the overlying epidermal epithelium when compared with the epithelial lining of tunnels . SignifiIL13, FGF21, COL3A1, VIM and EMT associated genes including SNAI2 and ITGB1 were significantly downregulated at Week 4 in lesional HS tissue compared with baseline (Week 0) (Spleen Tyrosine Kinase (SYK) inhibition with Fostamatinib at a dose of 100mg twice daily significantly modulates gene expression as measured by the Human Fibrosis V2.0 Nanostring gene expression assay. Fibroblast associated genes including (Week 0) . Pathway(Week 0) .This study presents novel gene expression and immunohistochemical data pertaining to the presence of fibroblast subpopulations and evidence of EMT in lesional tissue of HS. Fibroblast subpopulations in lesional tissue are significantly altered compared to non lesional tissue and healthy controls. Expansion of the SFRP2+ and CXCL12+ populations were identified with no significant alteration to the SFRP1+ population. Additionally, the severity of disease and presence of epithelialized tunnels is associated with these alterations to fibroblast subpopulations. Evidence of EMT was seen in HS lesional tissue with greater levels associated with epithelialised tunnels and Hurley stage 3 disease. SYK inhibition with fostamatinib for as little as 4 weeks resulted in reduced expression of fibroblast and EMT associated genes as measured by Nanostring gene expression assay. Pathway analysis of Nanostring data demonstrated enrichment of downregulated pathways pertaining to fibroblast proliferation and connective tissue growth factor production.findings in HS include gross expansion of the dermal layer, fibrosis and retention of inflammatory cells, all associated with function of SFRP2+ fibroblasts.Previously published functional annotation of fibroblast subpopulations in normal human skin identifies SFRP2+ (Type A) cells as functionally associated with dermal cell and extracellular matrix homeostasis as well as inflammatory cell retention. Additionally, SFRP2+ cells are involved in the development of tertiary lymphoid structures (TLO) . The sigCXCL12+ (Type B) cells contribute to immune surveillance and are associated with Th2 immune axis activation. This subset is particularly expanded in lesional tissue of atopic dermatitis . SFRP1+ Evidence of EMT has been reported in HS lesional tissue by Nelson and colleagues with reductions in E-cadherin in the epidermis . This haEMT can be induced in epithelial cells by inflammation and extracellular matrix alterations in both oncological and inflammatory settings , 27. AddThe use of whole genome RNAseq has several disadvantages when examining discrete gene expression pertaining to cellular subsets . Biases In-silico deconvolution provides a computational alternative to scRNAseq which, when utilized with a high-quality reference frame can provide insights into cellular proportions in whole RNAseq data whilst avoiding the biases present in scRNAseq. The quality of the deconvolution data is predicated on the baseline RNAseq data. Whilst whole tissue RNAseq is most appropriate for biomarker discovery, for identification of a priori differential gene expression, the combination of in silico-deconvolution of whole tissue RNAseq and Nanostring gene expression (confirmed with immunohistochemistry) provides a robust methodology for investigation of pre-defined signatures of fibroblast subpopulations.HS is in great need of novel therapies; however, our understanding of the disease pathogenesis and specific mechanistic action of novel therapies is incomplete. SYK is an important mediator of downstream signaling in monocytes, neutrophils and B cells but can also contribute to the reprogramming of fibroblasts . The posThis study was approved by the human research ethics committee of Sydney South West Area Health Service . A total of 20 participants underwent lesional and non lesional biopsies (6mm in diameter) based upon previously published definitions (Frew 2019b) . All bio2 transform the normalized counts. Differentially expressed genes were defined as fold change (FCH) of \u2265 \u00a61\u00b75\u00a6 and false discovery rate \u2264 0\u00b705.Total RNA was extracted using the Qiagen RNeasy kit as per standard protocol . RNA was sequenced using the Novaseq S4 Differential expression was estimated with DESeq2 and VST to logwww.pantherdb.org). Analyses were performed using whole tissue RNAseq data classification system (seq data as well seq data .https://cibersortx.stanford.edu/index.php) in order to estimate the abundance of fibroblast subpopulations in baseline lesional and non-lesional tissue using whole tissue RNAseq. The utilized fibroblast reference matrix was based upon previously published human skin scRNAseq data (GSE128169) with stratification into three major subtypes as defined by Ascension et al (https://doi.org/10.1016/j.jid.2020.11.028).In silico deconvolution was undertaken using the CIBERSORTx web application using the Human Fibrosis V2.0 gene panel. Raw data was processed using the Nanostring Nsolver (version 4.0.70) analysis software using quality control and normalization procedures derived from the NormqPCR R package as previously described . Differe2 of epidermis or dermis and measured using ImageJ v1.42 software. Differences between groups were analyzed using the Wilcoxon rank sum test for two variables and/or omnibus test using one-way ANOVA with adjustment for multiple comparisons was made using the Benjamini Hochberg procedure.IHC staining was undertaken using primary antibodies listed in Fostamatinib was administered as part of the Phase 2 clinical trial in moderate to severe HS (NCT0504698) . 100mg Bwww.r-project.org; R Foundation, Vienna, Austria) using publicly available packages (www.bioconductor.org). Means of each group and differences between groups were estimated using least-square means. Hypothesis testing was undertaken using the general framework for linear mixed-effect models of the limma package. tissue was considered a fixed factors while random effect related to subjects was included. Unsupervised clustering was performed with the Euclidean distance and average agglomeration. RNAseq data p-values were corrected for multiplicity using Benjamini\u2013Hochberg procedure.RNA sequencing statistical analysis was performed in R language Click here for additional data file.S2 Fig(TIFF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(XLS)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.S4 Table(XLS)Click here for additional data file.S1 File(DOCX)Click here for additional data file."} +{"text": "Patients undergoing revision total hip (THA) and knee (TKA) arthroplasties are at increased risk for surgical site infections (SSI) with prior infection representing a common indication for revision. Our aim was to evaluate patient characteristics and risk factors in patients with THA and TKA infection following revision arthroplasty.A single center retrospective review of patients with THA or TKA SSIs from 01/01/2017 to 11/01/2022 was performed utilizing National Healthcare Safety Network (NHSN) definitions and surgical revision codes. Index surgery was defined as the THA or TKA to which the SSI was attributed per NHSN guidelines. Patients meeting NHSN PATOS (present at time of surgery) definition were excluded. Pertinent demographics data, surgical histories and microbiology were collected and the determination of new vs. persistent infection was made based on microbiologic data.S. aureus 16 (34%), Enterococcus species 11 (23%), enterobacterales 10 (21%), and polymicrobial infections 10 (21%).Forty-seven cases were included with median age of 62, 45% male and 51% current/former smokers. BMI was \u226535 in 36% of patients, with 33% having diabetes mellitus and 13% were immunocompromised. Twenty-seven (57%) patients had a prior infection at the involved joint with 18/27 (67%) occurring within 1 year of the index surgery. Table 1 includes index surgery details and infection data. Median time to infection was 25 days. Leading pathogens were S. aureus was the most common pathogen identified but many cases involved less typical SSI pathogens. These results highlight the importance of focusing on patient specific risk factors, surgical infection prevention strategies and wound care, rather than additional pathogen specific antibiotics. While more study is needed to delineate SSI risk factors following revision arthroplasty, our study highlights the multitude of potential factors involved in these highly morbid infections.Over half of patients that developed revision THA/TKA SSI had a prior infection but less than 15% had a persistent infection. Most patients had multiple prior surgeries which may contribute to longer operative times and delayed wound healing. All Authors: No reported disclosures"} +{"text": "Liver metastases severely reduce the long term survival of colorectal cancer patients. Long non-coding RNAs (lncRNAs) CCAT1 and CCAT2 have previously been found to be associated with impaired patient outcomes in primary colorectal cancer. We aimed to elucidate the role of CCAT1 and CCAT2 in colorectal liver metastases.Total RNA was isolated from 97 human tissue samples of colorectal liver metastases and adjacent normal liver tissue. Gene expression analysis was performed by RT-qPCR and Multiplex ELISA and correlated with patient characteristics and survival. Gene expression, cancer cell migration, invasion, and proliferation were studied after siRNA-mediated knockdown of CCAT1, CCAT2, and MYC in metastatic colorectal cancer cell lines Colo205 and HROC277Met2.Elevated expression levels of lncRNAs CCAT1 and CCAT2, and their common target MYC in colorectal liver metastases were associated with prolonged progression-free survival after liver resection. High expression of CCAT1 was likewise associated with prolonged overall survival. Knockdown of CCAT1, CCAT2, and MYC resulted in increased migratory and invasive potential in metastatic colorectal cancer cell lines. Gene expression analysis revealed alterations in constituents of Wnt signaling following knockdown.Our findings demonstrate tumor-suppressive functions of lncRNAs CCAT1 and CCAT2 in colorectal liver metastases. They suppress Wnt signaling directly and indirectly through target gene MYC and might prevent further metastatic spread from colorectal liver metastases. Long non-coding RNAs (lncRNA) are a broad and heterogeneous group of molecules involved in transcriptional and translational gene regulation. Colon-cancer associated transcript 1 and 2 (CCAT1 and CCAT2) are two lncRNAs first described in primary colorectal cancer. Numerous reports have found a negative association of their expression levels in the primary tumor with patient outcomes . Both lnAround 25% of colorectal cancer patients will eventually face a diagnosis of liver metastases. Surgical resection remains the only potentially curative treatment option but is limited by patients\u2019 liver function and the extent of liver involvement . HoweverPatient samples were used from the tissue bank of the Surgical Oncology group at the Department of General, Visceral, and Transplantation Surgery at Heidelberg University Hospital. Only patient samples of colorectal liver metastases and corresponding physiologic liver tissue were included in this study. Our patient cohort compromised a total of 97 patients (for patient characteristics see 2 First Strand Kit (QIAGEN) for tissue samples and Improm-II-Reverse Transcription System for cell culture samples. Semi-quantitative RT-PCR was performed using Light Cycler \u00ae480 SYBR Green I Master and specific primers and transcribed into cDNA with RT primers . All proequation . mRNA exHuman Colo205 cells derived from malignant ascites of a colorectal cancer patient were purchased from Sigma-Aldrich . Human HROC277Met2 cells derived from a colorectal liver metastasis were purchased from CLS Cell Lines Service . Colo205 cells were cultured in RPMI with 2 mM Glutamine (Sigma-Aldrich). HROC277Met2 cells were cultured in DMEM/F12 with 3 mM Glutamine (Sigma-Aldrich). All cell culture media were supplemented with 1% Penicillin/Streptomycin (Sigma-Aldrich), and 10% heat-inactivated FCS (Sigma-Aldrich). Cells were grown at 37\u2009\u00b0C in a humidified 5% CO2 incubator.Transient knockdown was performed using a pool of siRNA sequences targeting human CCAT1, CCAT2, or MYC together with Lipofectamine RNAiMAX . AllStars Negative Control siRNA (Qiagen) served as control. Cells were incubated for 24 hours for RNA extraction and 48 hours for migration and invasion assay, proliferation assay, and protein extraction.Migration and invasion assay was performed by seeding cells onto a 8 \u03bcm pore transwell insert . The inserts were coated with Matrigel for invasion assay. Transfected cells were starved for 18 h in respective cell culture medium without FCS. Afterwards, medium in the lower chamber was replenished with 20% FCS serving as chemoattractant. Invading and migrating cells on the opposite side of the insert membrane were quantified after 24 h by dissolving cell-bound crystal violet (Sigma-Aldrich) in 10% acetic acid and measuring the optical density at 540 nm.Cellular proliferation following knockdown was assessed using BrdU Cell Proliferation ELISA (Roche) according to the manufacturer\u2019s instructions.Total protein was isolated from cell culture samples using RIPA lysis buffer with added protease inhibitor . Primary antibodies against human beta-actin , histone H3 , MYC , and beta-catenin were incubated overnight at 4\u00b0 C. After development with an HRP-conjugated secondary antibody , semiquantitative analysis was performed with ImageJ software . Densitometric measurement of protein bands was performed. MYC protein expression was normalized to Histone H3, and beta-catenin expression was normalized to beta-actin.Protein was isolated from a subset of human tissue samples described above using MILLIPLEX\u00ae MAP Lysis Buffer with added protease inhibitor (Merck Millipore) according to the manufacturer\u2019s instructions. Multiplex protein ELISA was performed using NF-\u03baB Signaling 6-plex Magnetic Bead Kit 96-well Plate Assay (Merck Millipore). Total \u03b2-Tubulin Magnetic Bead MAPmates\u2122 (Merck Millipore) were used for normalization of protein expression levels.Gene expression level analysis with semi-quantitative RT-PCR revealed increased expression of lncRNAs CCAT1 and CCAT2 in colorectal liver metastases compared to adjacent liver tissue . We likeWe next assessed associations of CCAT1, CCAT2 and MYC expression levels in colorectal liver metastases with patient outcomes. For this purpose, we separated our patient cohort into two groups using median lncRNA or mRNA expression as cut-off. Increased expression of lncRNAs CCAT1 and CCAT2 was associated with prolonged progression-free survival following surgical resection of liver metastases . ElevateBased on the clinical observations outlined above we hypothesized that CCAT1 and CCAT2 might exert tumor-suppressive functions in metastatic colorectal cancer cells via their target MYC. We selected two metastatic colorectal cancer cell lines, Colo205 and HROC277Met2 to test this hypothesis see .Knockdown of CCAT2 revealed divergent results among the chosen cell lines. In HROC277Met2, it resulted in a significant increase of migration and invasion , whereasPrompted by these findings, we assessed baseline expression levels of MYC in the two studied cell lines. MYC expression was strikingly higher in HROC277Met2 than in Colo205 on both the mRNA transcript and protein level . Based oWe likewise assessed the impact of CCAT1, CCAT2 or MYC knockdown on downstream effectors of Wnt signaling. Surprisingly, MYC knockdown caused a significant decrease of catenin beta-1 (CTNNB1) expression on mRNA and protein levels, despite MYC being a target of Wnt signaling .Cell culture experiments suggest that CCAT1 and MYC propagate a more invasive phenotype in metastatic colorectal cancer cells. Yet, these results are unlikely to be explained by altered proliferation, which was widely unaltered upon knockdown of CCAT1, CCAT2 or MYC. Knockdown of CCAT2 showed opposite results in Colo205 and HROC277Met2. This divergent phenotype could be explained by the different baseline expression levels of MYC in the two cell lines. The biologic output of MYC has been demonstrated to be determined by its baseline expression . TherefoInterestingly, MYC expression in HROC277Met2 is attenuated despite knockdown of CCAT1 on mRNA and protein level, whereas CCAT2 knockdown resulted in a significant decrease of MYC protein expression. This may suggest the existence of redundant mechanisms to promote MYC expression in this cell line.The divergent response of genes involved in Wnt signaling indicates that CCAT1 and CCAT2 possess distinct functions independent of MYC. The increase of TCF7L2 following knockdown of CCAT2 may indicate a regulatory feedback loop that preserves MYC mRNA expression. Furthermore, a trend towards increased beta-catenin protein expression was observed, as well. Similarly, CCAT1 knockdown showed a trend towards increased mRNA and protein expression of beta-catenin. In total, this would suggest increased Wnt signaling following knockdown of CCAT1 and CCAT2. However, previous reports have reported a contrary stimulating effect of CCAT1 on Wnt/beta-catenin signaling in oral squamous cell cancer .To our surprise, MYC knockdown resulted in decreased mRNA expression of beta-catenin and LEF1, despite MYC being a Wnt signaling target itself. This suggests that MYC expression generates a positive feedback loop with Wnt signaling. Furthermore, it has previously been reported that MYC-dependent LEF1 induction can increase beta-catenin expression .From a clinical point of view, CCAT1 and CCAT2 seem to exert protective functions in patients suffering from colorectal liver metastases. However, this should not be confused with a tumor-suppressive function of both lncRNAs but rather represents a different tumor biology. Surprisingly, the roles of CCAT1 and CCAT2 have previously been associated with unfavourable patient outcomes in various tumor diseases including primary colorectal cancer \u201327. In tIn summary, our study describes the protective effect of CCAT1 and CCAT2 mediated by their target MYC in colorectal liver metastases. Moreover, we were able to demonstrate the direct impact of CCAT1, CCAT2, and MYC on metastatic colorectal cancer cell migration by affecting targets of Wnt signaling, including the upregulation of PPARD. Finally, the divergent response of Wnt genes following knockdown demonstrates the existence of redundant mechanisms of action of CCAT1 and CCAT2 independent of MYC. It has previously been shown that colorectal metastases can seed metastatic cancer cells into the circulation . Based oS1 Fig(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file."} +{"text": "Here we present proteomic analysis comparing the modern broiler line, Ross 708, with the UIUC legacy line which is not selected for growth traits. Breast muscle proteome analysis identifies cellular processes that have responded to human directed artificial selection. Mass spectrometry was used to identify protein level differences in the breast muscle of 6-day old chicks from Modern and Legacy lines. Our results indicate elevated levels of stress proteins, ribosomal proteins and proteins that participate in the innate immune pathway in the Modern chickens. Furthermore, the comparative analyses indicated expression differences for proteins involved in multiple biochemical pathways. In particular, the Modern line had elevated levels of proteins affecting the pentose phosphate pathway, TCA cycle and fatty acid oxidation while proteins involved in the first phase of glycolysis were reduced compared to the Legacy line. These analyses provide hypotheses linking the morphometric changes driven by human directed selection to biochemical pathways. These results also have implications for the poultry industry, specifically Wooden Breast disease which is linked to rapid breast muscle growth.Chicken domestication began at least 3,500 years ago for purposes of divination, cockfighting, and food. Prior to industrial scale chicken production, domestication selected larger birds with increased egg production. In the mid-20 Initial efforts selecting for increased broiler mass led to adipose tissue accumulation, probably because the selection did not channel the increased level of nutrients to a particular tissue [The chicken has played an important role in human culture and nutrition since its domestication ,2. The mr tissue . UltimatComparative studies describing differences between selected and unselected (legacy) lines provide insight into the impact of artificial selection of species. Several studies have characterized the differences between modern broilers and legacy lines that have not been subjected to production level human directed selection \u201313. In oThe increase in the modern broiler\u2019s breast muscle mass, which almost reaches the hummingbird level, came at the expense of other tissues. For example, the normalized masses of the heart, spleen and brain are larger in legacy lines compared with modern broilers ,14,21. IIGF1) which stimulates muscle hypertrophy combined with reduced expression of myostatin (MSTN), an inhibitor of skeletal muscle hyperplasia [Our previous research involved comparing the breast muscle transcriptome of 6-day old (D6) modern and legacy lines . That coerplasia . These sHere we report a comparative study between modern Ross 708 and legacy UIUC breast muscle proteomes. The Ross 708 modern line is a commercial breed that is produced for human consumption. The breeding process for this line is proprietary information owned by Aviagen. As a result, a detailed description of the breeding process and the specific genetic background of Ross 708 is not publicly available. The UIUC legacy line birds were obtained from the University of Illinois, Urbana Campus. They are the progeny of a cross between males from the inbred New Hampshire and females from the inbred Columbian lines . The respectoralis major muscle [Animal raising, handling and sample collection methods were approved by the University of Delaware Institutional Animal Care and Use Committee (Permit Number: 2703-12-10). Six male UIUC and six male Ross 708 breast muscle samples were collected at day 6 (D6) post-hatch, frozen in liquid nitrogen and stored at -80\u02daC until processed for proteome analysis. All samples were taken from the posterior region of the left r muscle . We chosr muscle .For each of the muscle samples, six technical replicates were analyzed by mass spectrometry. In each case 50 mg of each muscle sample was subjected to differential detergent fractionation and 20 \u03bcg of each fraction was trypsin digested as previously described ,37. FollAn Ultimate 3000 (Dionex) high performance liquid chromatography system coupled with an LTQ Velos Pro (Thermo) mass spectrometer were used for peptide separation and mass spectrum acquisition. The U3000 was operated at a flow rate of 333 nl per minute and equipped with a 75 \u03bcm x 10 cm fused silica column packed with Halo C18 reverse phase material . Each peptide sample was separated using a 4 h gradient from 2% to 50% acetonitrile with 0.1% formic acid as a proton source. The column was located on a Nanospray Flex Ion Source (Thermo) and connected directly to a silica Nanospray emitter to minimize peak broadening. High voltage was applied using a stainless-steel junction between the column and the emitter. Scan parameters for the LTQ Velos Pro were one MS scan followed by 10 MS/MS scans of the 5 most intense peaks. MS/MS scans were performed in pairs, one using collision induced dissociation (CID) and the other using higher-energy collisional dissociation (HCD). Dynamic exclusion was enabled with a mass exclusion time of 3 min and a repeat count of 1 within 30 sec of initial m/z measurement.Spectrum matching programs X!tandem and OMSSDifferential expression of proteins between lines was performed pairwise using peptide elution profiles as described previously . PrecursGene Ontology (GO) enrichmeTranscript data discussed in this publication have been deposited in NCBI\u2019s Gene Expression Omnibus (GEO) and are accessible through GEO Series accession number GSE65217. Proteomics data is available from ProteomeXchange (PXD005288).In the comparison between D6 modern Ross 708 and legacy UIUC breast muscle, a total of 222 differentially expressed proteins were detected , with 173 enriched in modern samples and 49 enriched in legacy samples. Among the 222 differentially expressed proteins, 130 (57%) exhibited the same direction of enrichment as shown in an earlier transcriptome study S1 Tabl.Muscle from legacy birds were enriched for GO terms relevant to myofibers and energy production in striated muscle, along with vesicle transport . Muscle COX6A1), capping actin protein of muscle Z-line subunit beta (CAPZB), ribosomal protein S27 (RPS27), G protein subunit beta 4 (GNB4) and DnaJ heat shock protein family (Hsp40) member B11 (DNAJB11). 44 stress-associated proteins were elevated in the modern chicken muscle compared to the legacy samples. These function in a variety of stress related processes including acting as chaperones and cochaperones, responding to genotoxic stress, redox regulation, and protein degradation. Among these were four proteasome subunits that likely offset the increased burden of misfolded proteins due to the large amount of protein synthesis required for muscle hypertrophy. Nine heat shock proteins (HSPs) were also elevated in muscle from the modern line, all of which function as chaperones controlling the proper folding of client proteins. Five members of the TCP complex that acts as an ATP dependent chaperone controlling actin folding were enriched in the modern samples [In the comparison between modern and legacy muscle, 49 differentially regulated proteins were part of the stress response. Five stress response proteins were elevated in the legacy breast muscle including cytochrome c oxidase subunit 6A1 , troponin C (TNCC), tropomyosins (TMP1 and TMP4), myosin light chain 1 (MYL1), myomesin 2 (MYOM2), cytoskeletal keratins (KRT5 and KRT7), and FMR1 autosomal homolog 1 (FXR1). CAPZB, GSN, TNCC, TPM1 and TPM4 regulate the dynamics of actin filament assembly. MYL1 is a non-regulatory light chain that interacts with actin in generating contraction. MYOM2 is a structural component that stabilizes the M band of muscle and KRT5 and KRT7 are intermediate filaments components. FXR1 controls mRNA transport and translation [FXR1 in mice reduce limb musculature and result in early mortality [BIN1) is enriched in the modern line. BIN1 protein localizes to T-tubules [2+ signaling [BIN1 cause centronuclear myopathy, which causes muscle weakness and atrophy [Gene expression in legacy muscle showed enrichment for several myofibril proteins including gelsolin , seryl-tRNA synthetase (SARS), glycyl-tRNA synthetase 1 (GARS), alanyl-tRNA synthetase (AARS) and glutamyl-prolyl-tRNA synthetase 1 (EPRS). These would support the elevated translation required for the muscle hypertrophy seen in the modern line.The most striking contrast between modern broilers such as Ross 708 chickens and earlier breeds is the difference in both normalized and total breast muscle yield. Some of the increase in normalized breast muscle of modern broilers can be attributed to hypertrophy due to increased protein synthesis ,55. ThisHK) and muscle phosphofructokinase (PFKM) . HK1 catTKT) and phosphoribosyl pyrophosphate synthetase (PRPS1 and PRPS2). These enzymes function in the nonoxidative phase of PPP returning fructose 6 phosphate and glyceraldehyde 3 phosphate to glycolysis and providing ribose 5 phosphate as a precursor for purine and pyrimidine synthesis (The modern line breast muscle contained higher levels of the PPP enzymes transketolase (ynthesis .CS), aconitase 2 (ACO2), isocitrate dehydrogenases , dihydrolipoamide S-succinyltransferase , succinyl-CoA ligases (SUCLA2 and SUCLG1) and fumarate hydratase (FH). The components not elevated were succinate dehydrogenase complex flavoprotein subunit A (SDHA) and malate dehydrogenase (MDH). The enrichment for components of the TCA cycle indicates the central role of this pathway in the energy metabolism of the modern Ross 708 chickens compared with the legacy UIUC birds. In addition to providing coenzymes for oxidative phosphorylation, the TCA cycle provides intermediates for cataplerosis to be used as precursors for nucleotides, amino acids, and lipids. Anaplerosis could be supported by elevated levels of glutamic-oxaloacetic transaminase 2 (GOT2) that converts glutamate to \u03b1-ketoglutarate thus allowing for replenishing TCA cycle metabolites.The main pathway affecting energy production enriched in the modern line breast muscle was the TCA cycle . Most enACAD9), the rate limiting enzyme in the oxidation of fatty acyl CoA. ACAD9 is responsible for introducing trans double bonds into palmitoyl-CoA and initiating the beta-oxidation of this common lipid. Also enriched is enoyl-CoA delta isomerase 1 (ECI1) which is necessary for beta oxidation of unsaturated fatty acids. The transcript encoding hydroxyacyl-CoA dehydrogenase (HADH) is also elevated in the modern Ross 708 muscle. This enzyme acts repeatedly on lipids, sequentially removing two carbon units by oxidizing a 12-carbon fatty acid to acetoacetyl-CoA. Acetyl-CoA acyltransferase 2 (ACAT2), also enriched in Ross 708 breast muscle, oxidizes acetoacetyl-CoA to acetyl-CoA. The product of beta oxidation, acetyl-CoA, can then be metabolized via the TCA cycle to generate energy, or used in anabolic reactions to support rapid breast muscle growth.Enzymes involved in fatty acid beta oxidation are enriched in the modern birds compared with the legacy line . One enrALDH7A1), betaine\u2014homocysteine S-methyltransferase (BHMT), dimethylglycine dehydrogenase (DMGDH) and glycine amidinotransferase (GATM). ALDH7A1, BHMT and DMGDH form a pathway in the conversion of choline to sarcosine that is found in high concentrations in skeletal muscle. Choline is a precursor to phosphatidylcholine, an important component of cellular membranes. GATM is part of the pathway that mediates the interconversion between creatine and glycine. Creatine is also found at high concentration in muscle as it is important for energy storage as creatine phosphate. Further supporting creatine phosphorylation is creatine kinase B (CKB), which is elevated in modern breast muscle compared with the legacy line tissue. Three enzymes enriched in the modern line are involved in S-adenosylmethionine (SAM) production from methionine. These include methionine adenosyltransferase (MAT1A), aldehyde dehydrogenase 7 family member A1 (ALDH7A1), betaine\u2014homocysteine S-methyltransferase (BHMT), and adenosylhomocysteinase (AHCY). SAM functions as a universal methyl donor in biological systems.In addition to TCA enzymes affecting amino acid anaplerosis, modern skeletal muscle was enriched for enzymes affecting glycine, lysine, and methionine metabolism. Enzymes affecting glycine impact choline and creatine levels include aldehyde dehydrogenase 7 family member A1 (NDUFS7), cytochrome c reductases , cytochrome c oxidase subunit 6A1 (COX6A1) and ATP synthase F1 subunit gamma (ATP5C1), a component of the ATP synthase central stalk. Four mitochondrial components were found enriched in modern Ross 708 muscle: one subunit of the electron-transfer-flavoprotein (ETFA) along with inorganic pyrophosphatase 2 (PPA2). One NADH dehydrogenase (NDUFS4) and one component of the ATPase stalk, ATP synthase peripheral stalk subunit d (ATP5H) were also elevated in the modern muscle.Enzymes associated with the mitochondrial respirasome showed enrichment in the legacy line breast muscle. These enzymes include NADH:ubiquinone oxidoreductase core subunit S7 (COPB2), transmembrane p24 trafficking protein 2 (TMED2), and amphiphysin (AMPH). COPB2 functions in retrograde transport from the Golgi to the ER and is involved in recognition of specific vesicle cargo proteins [AMPH is enriched in neural tissue where it is involved in endocytosis through interaction with clathrin [AMPH autoantibodies produced in breast and lung cancer patients. SPS is characterized by muscle spasms, rigidity and hypertrophy that arise from the effect of the autoantibody on nerve cells that control muscles [CDC42), GDP dissociation inhibitor 2 (GDI2), Rac family small GTPase 1 (RAC1) and ras homolog family member A (RHOA).The legacy line breast muscle was enriched for proteins affecting vesicle transport including: COPI coat complex subunit beta 2 and bridging integrator 2 (BIN2) were enriched in the legacy line breast muscle. Elevated expression of GSN is associated with a decreased response to a variety of inflammatory stimuli [BIN2 levels are associated with decreased phagocytic activity [CTSB is secreted during neutrophil degranulation and degrades collagen upon release into the extracellular space [PRDX6 and RAC1 associate with and activate NADPH oxidase, a major source of reactive oxygen species [RAC1 [RHOA [WDR1) [Gene Ontology analysis identified 39 proteins that function in innate immunity. Two, gelsolin participA [WDR1) . It is rThe differences between modern broilers (Ross 708) and legacy lines (UIUC), arose from human directed selection leading to broiler\u2019s rapid growth, improved feed efficiency and increased breast muscle mass. Rapid growth is evidenced in the time from hatch to market. Typically, for the legacy line it takes 16 weeks for birds to reach market weight, while birds from the modern line take seven weeks that supports the nonoxidative part of the pentose phosphate pathway (PPP). The non-oxidative portion of the PPP provides precursors for nucleotide synthesis and feeds glucose metabolites back into glycolysis. Modern Ross 708 birds also express higher levels of phosphoribosyl pyrophosphate synthetases, which direct Ribose-5-Phosphate to nucleotide production. The elevated level of LDHB in the Ross 708 birds is expected to drive the lactate:pyruvate equilibrium towards pyruvate. This would retain pyruvate for further metabolism by the TCA cycle and limit the release of lactate for energy production by other tissues [The legacy line birds exhibited enrichment for ycolysis . PFKM ca tissues . Further tissues .Klf10 mouse knockout model of soleus muscle hypertrophy [The elevated levels of multiple enzymes of the TCA cycle in the Ross 708 breast muscle allows this pathway to meet demands of breast muscle hypertrophy . This isertrophy with simertrophy ,82. Furtertrophy .ACAD9 is particularly informative as this is the rate-limiting enzyme controlling lipid oxidation and elevated HADH activity also plays a role in skeletal muscle hypertrophy [ACAD9, HADH, and other enzymes involved in lipid oxidation indicates that modern Ross 708 birds are using lipid metabolism, in addition to the pentose phosphate and TCA pathways, to provide resources supporting expansion of the breast muscle.Several enzymes involved in lipid beta-oxidation are also elevated in birds from the modern line [PRDX1, PRDX4, PRDX6) and thioredoxin (TXN) play important roles in regulating oxidative stress. Gene Ontology analysis of our data also detected proteins associated with neutrophils in the mammalian immune system. Neutrophils are one of the earliest responders to inflammation and in birds the role of these phagocytic cells is filled by heterophils [Although these samples were obtained at day 6 post-hatch, there are already differences between the modern and legacy lines that have implications for the development of Wooden Breast Disease . The elePIT54) ,87, peroerophils . Histoloerophils . Furthererophils . Taken tIn support of our earlier transcriptome studies , these dS1 TableIncluded are proteins and gene accessions from NCBI and expression data supported by transcriptome expression is indic(DOCX)Click here for additional data file."} +{"text": "At our university medical center, we have well established programs for outpatient parenteral antimicrobial therapy (OPAT) and inpatient multidisciplinary antimicrobial infusions for patients with substance use disorder. Complex outpatient oral antimicrobial therapy (COpAT) uses oral (PO) in place of intravenous (IV) antimicrobials. Despite evidence supporting early IV to PO antimicrobial transition for serious infections, COpAT Programs are not widely incorporated in clinical practice. COVID-19 pandemic\u2019s impact on inpatient bed scarcity triggered COpAT\u2019s accelerated implementation at our institution. A Nov 2020 CEO email served as the catalyst and by Aug 2022 an Infectious Diseases (ID) Pharmacist was hired. By Oct 2022, an ID Physician at 0.5 FTE was assigned COpAT Medical Director. We hypothesized IV to PO antimicrobial transition increased as our COpAT Program developed.Early Discharge: Short course IV antimicrobials < 2 weeks followed by COpAT (n= 115); Traditional: Longer course IV antimicrobials \u2265 2 weeks followed by COpAT (n= 265). Demographic, clinical, and outcomes data were compared.Retrospective cohort study on 380 COpAT patients discharged Jan 2021 to Apr 2023 categorized in 2 groups: Proportion of patients in Early Discharge group increased significantly (Image 1). Traditional group had higher rates of injection drug use and bacteremia. While gram positive organisms predominated in both groups, Traditional group had a higher rate of monomicrobial infection (Image 2). Early Discharge group had a greater proportion of oral antimicrobial days and shorter length of stay but higher overall and COpAT-related 30-day readmission rates . Both groups avoided inpatient hospital days and had high clinical cure rates (Image 3). COpAT Program ROI was 10.9.COVID-19 pandemic accelerated development and maturation of our COpAT Program supported by 0.5 FTE ID Physician and 1.0 FTE ID Pharmacist. While readmission rates in both groups were low, possible association between shorter length of stay and increased readmission rates needs further evaluation. COpAT Program investment pays for itself in less than 2 months, mainly through inpatient days avoided.All Authors: No reported disclosures"} +{"text": "Interestingly,hdac6transcripts often associate with ciliated tissues, like the left-right organizer at neurula stage or the pronephros. In the embryonic skin, Hdac6 protein localizes to the cilia base, suggesting a functional link.Histone deacetylases (HDACs) are key posttranslational modulators of the proteome. We show that expression of Histone deacetylases (HDACs) are best known for their transcriptional regulation in eukaryotic cells, by controlling DNA accessibility for e.g. transcription factors via deacetylation of histone proteinsThe highly conserved and best analyzed HDAC6 (subclass IIb)predominantly localizes to the cytoplasm and was shown to serve as the major deacetylase of acetylated alpha-TubulinHdac6knockout mice are viable and fertileXenopus, with the exception of its contribution to the development of the visual system in swimming tadpoleshdac6mRNA in combination with its protein localization, which indicate an association with cilia.Surprisingly,hdac6throughout Xenopusdevelopment, we performed whole-mount mRNAinsituhybridization experiments.hdac6 transcripts are ubiquitously detectable in the animal hemisphere of stage 8 blastula embryos . Short of antibodies directed againstXenopusis in line with the previously reported, subtle LOF phenotypes in the corresponding embryonic tissues . Additionally, the observed localization of Hdac6 protein at the base of motile cilia points towards a more complex role of Hdac6 rather than exclusively regulating the axonemal acetylation of alpha-Tubulin.hdac6 expression in the LRO of the frog embryo argues for a functional contribution of Hdac6 in left-right patterning. Indeed, chemical modifier screens have previously identified HDAC inhibitors like Trichostatin A or Sodium butyrate as left-right defects causing agents whereas the mode of action remains elusivePublished data have implicated a role of Hdac6 in cilia homeostasis of different vertebrate species . The tissue specific developmental mRNA expression pattern inXenopusserves as an ideal model for the investigation of ciliary architecture and function in health and diseasehistone deacetylase 6in context of mucociliary epithelia and its functional role in the establishment of visceral left-right asymmetry.AsMaterials and methodsAnimal care and maintenanceXenopus laevisfrogs/embryos were approved by the Regional Government Stuttgart, Germany and performed according to German federal laws and regulations . Animals were staged according toHusbandry, handling and experimental manipulations ofXenopus laevis hdac6Xenopus laevis hdac6 cDNA in pCMV SPORT6 (clone IRBHp990G107D) vector was purchased from Source BioScience and linearized withNcoIfor antisense transcription with T7 polymerase (Promega).A full ORF clone fromRNA in situ hybridization and histological analysisin situ hybridization were fixed in MEMFA for 2 h at RT and later on processed following standard protocols suitable forXenopusembryosSp6orT7RNA Polymerase (Promega) from linearized plasmids. mRNA-ISHwas performed according toEmbryos dedicated forImmunofluorescenceXenopus laevisHdac6 Protein was detected by a commercially available antibody after overnight fixation (-20 \u00b0C) using Dent's fixative . Cilia located at the tadpole epidermis were visualized as described by Vick et al. 2009 After fixation embryos were treated according to standard IF proceduresEndogenous"} +{"text": "Scientific Reports 10.1038/s41598-023-43322-4, published online 03 October 2023Correction to: The original version of this Article contained errors in the legends of Figures 8 and 9.a) Mean differential polarization signals\u2009\u00b1\u2009standard error for the two tumor types. (b) and (c) Estimated scatterer size distribution for the chondrosarcoma and the osteosarcoma respectively. The nuclear and cellular size distribution estimated from histological analyses are normalized.Figure 8: (now reads:a) Mean differential polarization signal\u2009\u00b1\u2009standard error for the two tumor types. (b) and (c) Estimated scatterer size distribution for the osteosarcoma and the chondrosarcoma respectively. The nuclear and cellular size distributions estimated from histological analyses are normalized.Figure 8: (a) Chondrosarcoma estimated solution and its optimized linear combination of nucleus and cell size distribution. The cell scattering identified in the estimated solution accounts for 52%. (b) Osteosarcoma estimated solution and its optimized linear combination of nucleus and cell size distribution. The nucleus scattering identified in the estimated solution accounts for 69%.Figure 9: (now reads:a) Osteosarcoma estimated solution and its optimized linear combination of nucleus and cell size distribution. The nucleus scattering identified in the estimated solution accounts for 69%. (b) Chondrosarcoma estimated solution and its optimized linear combination of nucleus and cell size distribution. The cell scattering identified in the estimated solution accounts for 52%.Figure 9: (The original article has been corrected."} +{"text": "Staphylococcus aureus (MRSA). We aimed to quantify empiric vancomycin overuse among critically ill children with suspected sepsis.Vancomycin is among the most commonly prescribed antibiotics in US children\u2019s hospitals. When used empirically for sepsis, vancomycin primarily targets methicillin-resistant We performed a cross sectional study including all episodes of suspected sepsis in two tertiary care pediatric intensive care units (PICUs) between 11/2020 and 10/2022. Suspected sepsis was defined as collection of a blood culture and administration of one or more broad spectrum antibiotics within 12 hours. Episodes with prior broad-spectrum antibiotic use or transfers from an outside hospital within 7 days were excluded, as were episodes occurring within 14 days of a preceding sepsis episode. The frequency of MRSA among episodes of suspected sepsis was calculated based on the results of sterile site, respiratory, and wound cultures collected within 24 hours of the sepsis episode. MRSA history was defined as a positive clinical or screening culture or PCR within 6 months before the suspected sepsis episode. As a secondary analysis, we quantified the frequency of any vancomycin-requiring organism (Table 1).P< 0.01), as did the proportion with a history of MRSA , whereas identification of MRSA did not.We identified 1859 suspected sepsis episodes. Of these, 1135 (61%) received empiric vancomycin for a median of 3 days . MRSA was identified in 37 (2%) and any vancomycin-requiring organism was identified in 101 (5%). Among 1790 episodes with no history of MRSA, 1080 (60%) received empiric vancomycin and MRSA was identified in 17 (< 1%). MRSA was infrequent in community onset sepsis and in patients without central lines (Table 2). The proportion of episodes receiving empiric vancomycin differed between the two centers (70% vs 44%, Vancomycin was administered to 61% of critically ill children initiated on broad-spectrum antibiotics, but MRSA was identified in just 2%. Reducing empiric vancomycin overuse for community onset sepsis and in patients with no history of MRSA or central venous catheter is an actionable target for antimicrobial stewardship efforts in the PICU.Jason Newland, MD, Moderna: Grant/Research Support|Pfizer: Grant/Research Support"} +{"text": "Signal transducer and activator of transcription (STAT) signaling is critical for the pathogenesis of abdominal aortic aneurysms (AAAs). Though protein inhibitor of activated STAT3 (PIAS3) negatively modulates STAT3 activity, but its role in AAA disease remains undefined.\u2212/\u2212) and wild type (PIAS3+/+) male mice via transient intra-aortic elastase infusion. AAAs were assessed by in situ measurements of infrarenal aortic external diameters prior to (day 0) and 14 days after elastase infusion. Characteristic aneurysmal pathologies were evaluated by histopathology.AAAs were induced in PIAS3 deficient and smooth muscle cell loss (media score: 3.0) than those in PIAS3+/+ mice (media score: 4 for both elastin and SMC destruction). Aortic wall leukocyte accumulation including macrophages, CD4+ T cells, CD8+ T cells and B cells as well as mural neovessel formation were significantly reduced in PIAS3\u2212/\u2212 as compared to PIAS3+/+ mice. Additionally, PIAS3 deficiency also downregulated the expression levels of matrix metalloproteinases 2 and 9 by 61% and 70%, respectively, in aneurysmal lesion.Fourteen days following elastase infusion, aneurysmal aortic diameter was reduced by an approximately 50% in PIAS3PIAS3 deficiency ameliorated experimental AAAs in conjunction with reduced medial elastin degradation and smooth muscle cell depletion, mural leukocyte accumulation and angiogenesis. Inflgression \u20135. Inhibgression \u20138.Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling critically regulates inflammatory responses , 10. It Excessively activating JAK/STAT signaling leads to dysregulated immune responses and thus tissue damage. Several endogenous inhibitory proteins, such as protein inhibitor of activated STAT (PIAS) and suppressor of cytokine signaling (SOCS), are evolved to limit overwhelming inflammation due to augmented JAK/STAT activity. We previously showed that PIAS3 was reversely associated with atherosclerotic progression and inhibited inflammatory responses and smooth muscle cell (SMC) proliferation . AAA as \u2212/\u2212) mice on C57BL/6 genetic background were created in Cyagen Biosciences (Suzhou) Inc using the CRISPR/CAS9 technique. Briefly, the exons of 2\u20139 were selected as target sites, and two gRNAs with Cas9 mRNA were microinjected into zygotes for PIAS3 deficient mouse generation (via PCR genotyping followed by DNA sequencing. Homozygotes (PIAS3\u2212/\u2212) and wild type (PIAS3+/+) littermates were screened and used for all experiments. The use and care of animals as well as all experimental procedures were reviewed and approved by the Laboratory Animal Administration Committee of Xi'an Jiaotong University (No. 2019\u20131178). All information on animals and related reagents were detailed in PIAS3 gene (NM_001165949) is located on mouse chromosome 3 with fourteen exons . PIAS3 dneration . F0 foun\u03b2-actin were obtained from Cell Signaling Technology and TransGen Biotech Co., Ltd , respectively.For PCR screening, tail tip samples were collected from <3 weeks old mice and genomic DNA was extracted by proteinase K digestion. Genotyping PCR primers were AAACAAGACTAAAGGAGTATGGGC (sense 1), TAGAGGAAGGGGAAGGGAACTAAG (antisense 1) and CTCAGACACTCGGAAACTCATC (antisense 2). For quantitative reverse transcription PCR (qRT-PCR) and Western blotting analyses, total RNA and proteins were extracted from aortas. The PIAS3 primers for qRT-PCR analysis were sense: ACCAAGAATGGAGCTGAGCC (sense) and TCTGGATTCCGGATCCCCTT (antisense). Antibodies against PIAS3 and \u2212/\u2212 and PIAS3+/+ mice by transient luminal infusion of porcine pancreatic elastase in controlled infrarenal aortic segment as previously reported and served the baseline level. On day 14 after PPE infusion, infrarenal aortic diameter for each mouse was measured by the same procedure prior to sacrifice and aorta was then harvested. A mouse with a more than 50% aortic dilation over the baseline was considered aneurysmal .\u03b1-actin antibody followed a standard biotin-streptavidin peroxidase procedure. Biotinylated donkey anti-goat IgG antibody and streptavidin-peroxidase conjugate were obtained from the Jackson ImmunoResearch Laboratories Inc., West Glove, PA, United States). AEC substrate kit for color development was purchased from Vector Laboratories Inc., Burlingame, CA, United States. Aortic medial SMC depletion was graded on a scale of I (mild) to IV (severe) as reported previously . Frozen sections were stained with hematoxylin and eosin (H & E) and Elastic van Gieson (EVG) stains, respectively, for the assessment of general morphology and elastin integrity. Aortic media elastin degradation was graded as I (mild) to IV (severe) on EVG-stained sections as previously reported \u201320. To e+ T cells, CD8+ T cells and B cells (B220) , CD4s B220) . Section . SectioMatrix metalloproteinases (MMPs), particularly MMP2 and MMP9, contribute to the pathogenesis of AAAs. MMP2 and MMP9 were assessed by immunostaining using goat anti-mouse polyclonal antibodies against MMP2 (AF1488) and MMP9 (AF909) (Cite your JIR article). The expression levels were quantitated as the positively stained area per ACS using the image analysis software .Angiogenesis was analyzed by immunostaining with rat anti-mouse CD31 monoclonal antibody previously described. The neovessels number were counted under the microscope and reported as CD31-positive vessels per ACS , 21.t and nonparametric Mann-Whitney tests were used for normally and nonnormally distributed data, respectively. Two-way ANOVA followed by Sidak's multiple comparisons test was used for testing statistical difference for aortic diameters among groups. Statistical significance level was set at p\u2009<\u20090.05.Prism 9.0 software was used for all statistical analyses. Continuous variables were expressed as mean and standard deviation or media with interquartile . Student's \u2212/\u2212 as compared to PIAS3+/+ mice.As illustrated in +/+ and PIAS3\u2212/\u2212 mice. Experimental AAAs were successfully induced in PIAS3+/+ mice pathway plays a vital role in the development of AAAs (We previously found that PIAS3 was negatively associated with the JAK2/STAT3 activation and inflammatory responses in macrophages during atherosclerosis . Unexpec protein . HIF-1\u03b1/ of AAAs \u201343. ThusIn conclusion, the present study demonstrated that genetic PIAS3 deficiency attenuated experimental AAAs in association with reduced medial elastin degradation, SMC depletion, leukocyte infiltration and aortic wall angiogenesis. Due to the limited aneurysmal tissues, the functions of MMPs and STAT3 were not determined in the present study. Further studies on the mechanism of PIAS3 regulating AAA will be conducted in the future."} +{"text": "Lower extremity infections (LEIs) of diabetic patients poses a challenge to addressing over-utilization of broad spectrum antibiotics (BSA). To guide the development of educational and behavioral initiatives, we employed ASP quality improvement surveillance practices to explore podiatric-surgery collected bone cultures (BC), pre-procedural tissue cultures (PTC) and results of resection margin pathology.Washington DC VAMC is an urban medical center with 220 acute and LTC beds with acute and outpatient surgical Podiatry Service and supported by an active ASP (PharmD/MD/NP). Between 2020-2022 positive BCs collected in podiatric LEI procedures for presumed osteomyelitis were reviewed by ASP with CDSS and EMR (VISTA_CPRS). A positive post-resection margin (Margin+), empiric antibiotic selection and microbiology including regimen altering organisms such as MRSA and P. aeruginosa (PSAR) were collected.Diabetic individuals had 94 positive BCs in 296 procedures among nearly 2,000 total cultures. Subjects were predominately male (97%), mean age 68 \u00b1 8.7 (45-93y); and nearly a third had > 1 BC. Intraoperative BCs made up 79% of episodes and half were Margin+. Subjects had high rates of PVD (45%), insulin-dependence (49%) and HbA1c > 10 mg/dL (21%). Empiric BSA selection consisted mainly of MRSA and PSAR coverage with no deaths < 30d from BC. Among BC with PTC, 61% had partial or complete correlation in microbiology; MRSA and PSAR incidence were the same if PTC:BC findings were concordant or not. PTC:BC microbiologic non-concordance vs concordance for Margin+ was not significantly different between the groups and among PTC:BC concordant samples, PSAR rates trended higher with no difference in MRSA incidence.Providers have limited information in predicting LEI results in advance of path and micro. Empiric BSAs remains a challenge for ASPs. Less than half of patients had a BC finding that would have been predicted by the PTC and for those with a concordant prior to procedure sample, the finding of organisms that directed the empiric selection were not reflective of final cultures. There are opportunities for ASP interventions that address empiric selection in diabetic LEIs.All Authors: No reported disclosures"} +{"text": "In December 2021, Evusheld, a cocktail of 2 monoclonal antibodies (mAB), was granted EUA for COVID-19 pre-exposure prophylaxis (PrEP) of immunocompromised patients (Table 1). The EUA was updated June 2022 to recommend repeating a dose every 6 months for patients needing ongoing protection. However, EUA was revoked in January 2023 due to identified resistance of Omicron lineage. Clinical trials about a new mAB for PrEP are continuing. Real-world data on the effectiveness of Evusheld may help develop strategies for the administration of new mAB.ENDURANCE is a complex retrospective study to assess Evusheld's effectiveness by evaluating and comparing treatment outcomes among 4 groups of patients classified based on the timing of treatment and the number of repeated Evusheld treatments . This pilot study describes treatment outcomes in the first analyzed group (gr. A) treated before Omicron development.Table 1EUA criteria for Evusheld use, used as inclusion criteria for our study. * Patients with moderate to severe immunosuppression due to a medical condition or receipt of immunosuppressive medications and may not mount an adequate immune response to COVID-19 vaccination.Figure 1ENDURANCE Study design. 459 patients who received the initial dose of Evusheld were followed for 180 days . Patients who received repeated treatment were followed for 180 days following an additional dose while patients who did not receive repeated treatment were followed for an additional 180 days . Primary outcome is defined as confirmed symptomatic COVID-19 infection. We hypothesize a significantly lower rate of COVID-19 infections in the period before Omicron emergence (Groups A and B), disregarding if patients received repeated doses of Evusheld (Group A\u2019) or not (Group B\u2019). A Pilot Study describes the treatment outcomes in group A.We conducted a retrospective chart review of patients treated with Evusheld under EUA between January 18 and June 21, 2022, to assess their response to treatment by evaluating outcomes listed in Figure 2. Patients were classified in tiers based on the degree of underlying immunosuppression (Table 2). In addition, COVID-19 anti-Spike antibodies were requisitioned before treatment.Figure 2Evaluated outcomes. Primary outcome was positive COVID test at any point during the study period. Secondary outcomes were measured through the number of all-cause ED visits, hospitalizations, ICU admissions and deaths within 30 days from the positive COVID-19 test.Table 2Tiered system for prioritizing patients based on severity of immunosuppression should resources be scarce.From 151 patients, the majority were vaccinated (95%), seropositive (66%), and classified as Tier 1 at the time of treatment (58%) (Table 3). All seronegative patients (39) were vaccinated . 16/159 (11%) developed symptomatic COVID-19 infection within 180 days from treatment (mean 119.8 days); 10/16 patients received antiviral treatment outpatient (Table 4). None visited ED, were hospitalized, or died.Table 3Patients\u2019 vaccination status and anti-Spike antibody (serology) results per tiers. *All seronegative patients were vaccinated; unvaccinated patients (8) were either seropositive (5) or with unknown serology (3).Figure 3Serum anti-Spike antibody results in vaccinated and unvaccinated patients. All seronegative patients (n=39) were vaccinated. Unvaccinated patients were either seropositive (n=5) or with unknown serology results.Table 4Characteristics of COVID-19 positive patients, compared to patients not diagnosed with COVID-19.Our pilot data suggests that Evusheld may be effective against susceptible strains, as demonstrated by the low number of symptomatic infections, which all were mild and did not require ED or hospital utilization . A tier system may help prioritize patients in scarce resource conditions. Further studies are needed to identify risk factors for severe COVID in immunocompromised patients and understand their immunological response to vaccines and natural infection to create the most helpful tier system.Figure 4Treatment outcomes. 10% (16/151) of Evusheld recipients developed mild symptomatic COVID-19. Number of all cause ED visits, hospitalizations, and deaths within 30 days from testing positive for COVID-19 was 0, suggesting possible beneficial effect of Evusheld use in immunocompromised population.All Authors: No reported disclosures"} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-023-41879-8, published online 22 September 2023Correction to: The Acknowledgements section in the original version of this Article was incomplete.\u201cThis work was supported by NASA Exobiology awards 80NSSC19K0469 (MIT) and 80NSSC22K0189 (Georgia Tech) to CEC, and the Human Frontiers Science Program grant RGY0066/2018 to AAB.\u201dnow reads:\u201cThis work was supported by NASA Exobiology awards 80NSSC19K0469 (MIT) and 80NSSC22K0189 (Georgia Tech) to CEC, and the Human Frontiers Science Program grant RGY0066/2018 and the European Research Council Grant no 818602 to AAB.\u201dThe original Article has been corrected."} +{"text": "The retraction has been agreed following an investigation into concerns raised by a third party, which revealed inappropriate duplications between this and other published articles [The above article, published online on 14 July 2018 in Wiley Online Library (articles , 4. Thus"} +{"text": "Mycobacterium tuberculosis (Mtb) in patients with difficulty in expectorating. However, high mycobacterial culture contamination rates and Xpert MTB/RIF Ultra test error rates on stool samples have limited its use. OMNIgene SPUTUM (OM-S) is a sample transport reagent with characteristics of sputum decontamination while maintaining viable Mtb. We evaluated the impact of OM-S on Mtb diagnostic yield from stool using smear microscopy, Xpert MTB/RIF Ultra, and culture among presumptive TB patients.Stool is a potential sample for diagnosing Mtb using concentrated smear fluorescence microscopy (CFM), Xpert MTB/RIF Ultra (Xpert) and Mycobacteria Growth Indicator Tube (MGIT) culture. Sensitivity, specificity, and predictive values for each test were calculated against sputum MGIT culture as the reference standard.Paired stool and expectorated sputum samples were collected from consecutive Ugandan adults undergoing diagnostic evaluation for pulmonary TB between June 2018 and June 2019. Stool was divided into 2 portions: one was homogenized in OM-S (OM-S stool) and the other in PBS (PBS stool) as control. Both sputum and processed stool were tested for Of the 200 participants, 120 (60%) were male, 73 (37%) were HIV positive (median CD4 120 cells/uL (IQR 43\u2013297)) and 128 (64%) had confirmed pulmonary TB by sputum MGIT culture. Seven (4%) OM-S stool Xpert samples reported errors while 47 (25%) and 103 (61%) were contaminated on OM-S stool MGIT and PBS stool MGIT, respectively. OM-S stool MGIT was able to accurately diagnose 56 of the contaminated PBS stool MGIT samples compared to only 5 of the contaminated OM-S stool MGIT samples diagnosed by PBS stool MGIT.Sensitivity 89% (83\u201394) for OM-S stool Xpert was higher compared to that of OM-S stool MGIT 60% (51\u201369) and PBS stool MGIT 42% (32\u201352). Specificity (95%CI) 91% (82\u201397) was also higher for OM-S stool Xpert compared to OM-S stool MGIT 64% (51\u201375) and PBS stool MGIT 26% (16\u201338).Mtb diagnostic yield and reduce rates of indeterminate results when tested on Xpert and MGIT culture. The method may thus be of value in Mtb detection among patients with difficulty to expectorate.Stool processed with OM-S showed potential to improve Tuberculosis (TB) is the leading cause of death from a single infectious agent worldwide, responsible for 1.4 million deaths annually . Early aMycobacterium tuberculosis (Mtb) bacilli or DNA may pass intact through the gastrointestinal tract into their stool. Prior studies have identified both live Mtb and dead Mtb DNA in stool using standard TB tests .A smear of 1-2cm was prepared and air dried using 1ml of the homogenized stool or sputum sample. Smears were stained with 0.1% Auramine-O , decolorized using 3% acid alcohol, and counterstained using 0.5% potassium permanganate before examination at 400X magnification using fluorescent microscopy. Results were recorded in the format outlined by the Clinical and Laboratory Standards Institute grading standards.One volume (mL) of the OM-S stool or sputum sample was mixed with two volumes (mL) of the Xpert sample reagent, then loaded into cartridges and tested according to the manufacturer\u2019s recommendations. Semi-quantitative results were recorded. Stool processed with OM-S was the only type tested on Xpert.M. tuberculosis complex from nontuberculous mycobacteria. Time-to-culture-positive (TTP in days) was noted from the BACTEC MGIT 960 System. Results were reported according to International Union Against Tuberculosis and Lung Disease guidelines .Of the 222 eligible patients, 10 were excluded at enrolment and 12 at data analysis due to reasons stated in Mtb, 76 (38%) were Xpert negative and 7 (4%) were invalid errors . Additionally, 50 (43%) of the 116 stool Xpert positive results were graded as \u201clow positive\u201d. Only 1 result was identified as rifampicin resistant and 7 had indeterminate rifampicin resistant status. Four of the 7 samples that were indeterminate rifampicin resistant were also reported as \u201cvery low positive\u201d, 1 low positive, and 2 were \u201ctrace positive\u201d. OM-S stool Xpert detected an additional 35 and 55 Mtb positive cases compared to OM-S stool MGIT and OM-S stool microscopy, respectively.Of the 199 OM-S stool samples tested on Xpert, 116 (58%) were positive for . Among the sputum smear negative PTB cases, sensitivity (95%CI) was highest for OM-S stool Xpert 65% (43\u201383) compared to stool MGIT at 39% (14\u201368) and stool smear at 12% (3\u201331) . Of the 47 contaminated OM-S stool MGIT samples, OM-S stool Xpert was able to identify accurately 21/25 (84%) samples as positive and 18/21 (86%) as negative when sputum MGIT was used as the reference. One sample was both culture-contaminated and had Xpert error.Sensitivity (95%CI) and specificity (95%CI) of OM-S stool Xpert were 89% (83\u201394) and 91% (82\u201397), which were lower compared to sputum Xpert sensitivity and specificity of 95% (90\u201399) and 90% (86\u201396), respectively . Stool samples had a higher average MGIT time-to-positivity 12 days (IQR 8\u201318) compared to 7 days (IQR 4\u201311) for sputum MGIT culture. Among the non-contaminated stool and sputum samples, stool MGIT identified 74/99 (75%) PTB positive cases and 44/69 (64%) PTB negative cases, respectively. Sensitivity (95%CI) and specificity (95%CI) of stool MGIT after exclusion of contaminated results were 75% (65\u201383) and 92% (80\u201398), respectively . When contaminated samples were included in the denominator, sensitivity of stool MGIT was reduced from 75% to 60% and specificity from 92% to 64%, respectively, providing a conservative estimate of these tests . We noted that OM-S stool MGIT was more likely to be positive with an increasing level of stool smear grade. For example, the highest proportion of positive stool MGIT results were identified among participants with smear grades \u201c3+\u201d (4/4), followed by (13/17) among those with smear grade \u201c2+\u201d compared to (33/91) among those with smear negative .We observed a 10-fold higher contamination rate for stool MGIT 47/194 (25%) compared to sputum MGIT 4/200 (2%) We observed that of the 78 positive OM-S stool MGIT samples, only 4 samples tested negative by sputum MGIT; of these 3/4 corresponding sputum samples were negative on sputum Xpert, and 4/4 corresponding sputum samples were negative on sputum smear. We also noted that 19 OM-S stool MGIT results were negative, but were positive by OM-S stool Xpert. Of these, 17/19 (89%) had already been confirmed positive by sputum MGIT. Also noted were the 11 samples which were OM-S stool culture negative but stool smear positive, 9 of these were sputum MGIT positive.. A significant difference in contamination rates was observed among samples processed by PBS 103/169 (61%) compared those processed by OM-S processed stool 47/194 (24%) . Both methods agreed on 15 (9%) negative, 36 (21%) positive and 31 (19%) contaminated samples. Fifty-six (29 negative and 27 positive) of the PBS stool MGIT contaminated samples, were accurately diagnosed by OM-S stool MGIT while PBS stool MGIT could only diagnose accurately 5 (3 negative and 2 positive) samples of contaminated OM-S stool MGIT samples when compared to sputum MGIT as reference. With inclusion of contaminated samples in the analysis a higher sensitivity 60% (95%CI 51\u201375) among OM-S processed samples compared to 42% (95%CI 32\u201352) was observed . A similar trend was observed with the specificity . Comparative analysis of sensitivity and specificity after exclusion of contaminated samples was not done due to high levels of contamination among the PBS processed samples and therefore high risk of misclassification bias.OM-S processed stool samples had a higher positivity rate 78/194 (40%) compared to PBS processed stool 45/169 (25%) . Specificity of OM-S stool smear at 97% (95%CI 90\u2013100) was similar to that of sputum smear 97% (95%CI 90\u201399) however lower than that of PBS-stool 99% (95%CI 92\u2013100) . Compared to stool Xpert and OM-S stool MGIT, stool smear showed the lowest levels of sensitivity . Both stool-smear and stool MGIT agreed on the positivity of 45 stool samples which were also confirmed positive by sputum MGIT. We observed that of the 47 contaminated stool MGIT samples, stool smear identified only 3/26 (12%) as positive and 21/21 (100%) as negative accurately basing on sputum MGIT as reference.Out of the 128 bacteriologically confirmed PTB cases, OM-S stool smear microscopy identified 57 (45%) accurately. Sensitivity of OM-S stool smear 45% (95%CI 36\u201354) was lower than 50 (95%CI 40\u201360) and 80% (95%CI 71\u201386) for PBS-stool and sputum smear, respectively Mtb diagnosis among patients who cannot easily provide sputum [Mtb yields, molecular diagnostic test errors and high rates of culture contamination [Mtb yield.Our stool processing method was able to diagnose PTB with a higher stool MGIT culture sensitivity and a within range sensitivity of stool Xpert compared to previous methods. We also observed a lower stool culture contamination rate compared to our PBS processed stool control. Previous studies have shown stool samples as potential alternative to respiratory specimens for e sputum ,9,22,23.Mtb bacteria caused by the harsh acidic conditions of gastrointestinal tract system in addition to stool sample processing methods compared to Mtb DNA identified by Xpert MTB/RIF whether dead or viable. These findings further emphasize Xpert MTB/RIF as currently the most suitable test for diagnosis of stool Mtb hence the recent WHO recommendation to use stool Xpert for initial diagnosis of TB among children [Mtb. Due to the low numbers of sputum smear negative PTB patients in our study, we cannot rule out the possibility of random bias in our findings. We therefore recommend evaluation studies of our stool processing method on Xpert among a bigger sample size of sputum smear negative PTB patients.Our findings demonstrated that when using our processing method, Xpert MTB/RIF was still the most sensitive standard diagnostic TB test on stool samples. Our sensitivity 89%) was higher than some of previous studies 9% was hi\u201315,24,25children . Our stoStool Xpert error results are due to inhibition of DNA Taq polymerase enzyme by faecal inhibitors leading to failure of Xpert PCR amplification ,28. PrioMtb DNA in stool is dependent on patients swallowing of enough volume of Mtb infected sputum into the GI tract compared to using of sputum with high bacterial load directly from the lungs for Mtb diagnosis. Our hypothesis is supported by our findings which show that the higher the sputum microscopy smear grade the higher the likelihood for a positive stool Xpert and MGIT. Because we collected on spot stool samples on the same day of collecting sputum samples, we think some of the PTB positive patients enrolled in the study may not have swallowed enough sputum by the time we accessed the stool sample.Sputum Xpert was still more sensitive (94%) compared to our stool Xpert (89%). This difference may be explained by the fact that detecting A lower sensitivity was observed for our OM-S processed (46%) stool microscopy as compared to other stool index tests. This was consistent with sensitivity of sputum microscopy compared with other sputum index test as reported by others ,31. Our Mtb test [Mtb in hard to reach areas where molecular assays like Xpert may not be available.The ease with which microscopy can be conducted makes the method the most available Mtb test and despMtb gold standard test and the most sensitive in the diagnosis of pauci-bacillary TB [Mtb culture performance on stool therefore would be important for clinical management. High level stool culture contamination rates [M. tuberculosis bacilli is a major challenge in utilization of stool culture for Mtb diagnosis. Our stool processing method had lower culture contamination rates compared to our control and studies by [Mtb viability during stool processing. Using our findings, we believe our OM-S based processing protocol may have potential to optimize and balance these two characteristics despite a higher time to culture positivity than sputum.Culture is the llary TB ,34. Undeon rates ,23 causeudies by ,23. We budies by were resudies by ,12,22 reMtb sensitivity for any stool culture study among adults with PTB compared to the next highest sensitivity of 54% by Amel El Kh\u00e9chine, et. al. [Mtb in when using stool culture for diagnosis despite the harsh acidic environment of stool.Our OM-S processed stool culture sensitivity 76%) after exclusion and (60%) with inclusion of contaminated samples in analysis respectively to our knowledge is this highest 6% after Stool Xpert however was able to accurately diagnose 85% the culture contaminated samples when we used sputum MGIT as a reference. Stool MGIT on the other hand could only diagnose accurately 57% (4/7) of the stool Xpert errors stool. The relationship may be an indication that with our stool processing method clinicians may benefit in using both methods with stool culture being opted in high-risk patients who have had Xpert errors or a negative stool Xpert result in addition to patients who critically need drug susceptibility testing results to rule out or treat Multi drug resistant TB. We did not find any studies done to assess benefit of this conditional utilization of both methods. This study did not have the power to investigate this objective and thus we recommend future studies to evaluate the benefit and feasibility of using both stool Xpert and culture among the categories of patients mentioned above.M. tuberculosis from stool sample of sputum producing bacteriologically positive patients when tested on standard of care M. tuberculosis tests. Our findings provide insightful information of a potential TB diagnostic stool processing method for future studies in paediatric population and other patients with difficulty to expectorate. During analysis, we did not adjust for the possibility of extra-pulmonary TB such as GI or lymph node TB to which some of our results may be attributed especially in Mtb positive stool samples which did not have any positive corresponding sputum tests. A single Rifampicin resistant sample we identified was not tested for DST and thus could not understand the utility of our method to diagnose RIF resistance when using stool Xpert. Due to limited study funds, we had logistical challenges to procure enough Xpert cartridges to test stool homogenized in PBS on Xpert. We thus did not have a comparator for our stool processing method tested on Xpert. This limited our ability to confidently attribute stool Xpert outcomes on our processing method. We however anticipate that publication of our findings will demonstrate the potential and importance of this additional test during similar future studies.One of the limitations of our study is that we did not include paediatric or severely sick adult patients both of whom would best benefit from this study. Before applying our stool processing method to paediatric or severely sick patients we needed to validate if our method can detect Mtb diagnosis in populations with difficulty in sputum production using both stool molecular and culture tests. With WHO now recommending use of stool Xpert for diagnosis of PTB in children less than 10 years old, focus should now be on how best to process stool for culture for better Mtb yields with less contamination. We think our method has attempted to balance both these characteristics although the culture contamination rates are still very high compared to sputum culture and thus, we cannot suggest replacing our stool culture with sputum culture in the diagnosis of PTB. We however believe our method still has a role in diagnosis of high-risk patients such as paediatric patients at risk of MDR-TB and HIV+ patients who cannot benefit from stool Xpert. We recommend future studies of this protocol to be done directly among paediatric cases, drug resistant and severely sick patients who cannot produce sputum.Our stool processing method demonstrated high potential to improve S1 File(XLSX)Click here for additional data file.S2 File(DOCX)Click here for additional data file."} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-023-41908-6, published online 22 September 2023Correction to: The original version of this Article contained errors in the Results and Discussion sections.In the Results section, the sentence:\u201cParticipants aged less than 40\u00a0years old using weight loss agents showed increased risks of elevated liver enzymes , and significant associations were found between elevated liver enzymes and the use of traditional medicines and supplements .\u201dnow reads:\u201cParticipants aged less than 40\u00a0years old using weight loss agents showed increased risks of elevated liver enzymes , and significant associations were found between elevated liver enzymes and the use of weight loss medications with prescription and supplements .\u201dIn addition, in the Discussion section, the sentence:\u201cThe association between the use of traditional herbal medicines and elevated liver enzymes was significant in men and young people.\u201dnow reads:\u201cThe association between the use of traditional herbal medicines and elevated liver enzymes was significant in men.\u201dThe original Article has been corrected."} +{"text": "SABA prescriptions (number of canisters per year) were defined as high (\u22653) or low (1\u20132). Association of SABA prescriptions with HCRU was assessed by negative binominal model adjusted for covariates. The UK unit costs from the NHS were applied to estimate total healthcare costs (2020). Medication costs were based on the annual average number of canisters per year per patient.Overall, 186 061 patients with SABA prescriptions were included, of whom 51% were prescribed high SABA. Total annual average costs (HCRU and medication) were 52% higher in the high SABA group versus the low SABA group (\u00a32 256 091 per 1000 patients/year versus \u00a31 480 640 per 1000 patients/year). Medication costs accounted for the majority of asthma-related costs. Across both groups, most HCRU costs were for non\u2013exacerbation-related primary care or hospital outpatient visits. The annual average HCRU cost difference for high SABA versus low SABA was the greatest for hospitalisations and exacerbation-related primary care visits . Asthma-related HCRU extrapolated to the broader UK asthma population was \u00a3108.5 million per year higher with high SABA versus low SABA.High SABA versus low SABA prescriptions are associated with higher asthma-related HCRU costs."} +{"text": "The present study indicates PMV to significantly correlate to elevated early postoperative mortality rates following stabilization surgery for PSD. These results might entail further scientific efforts to investigate PMV as a so far underestimated negative prognostic factor in the surgical treatment of PSD.Surgical procedures with spinal instrumentation constitute a prevalent and occasionally highly indicated treatment modality in patients with pyogenic spondylodiscitis (PSD). However, surgical therapy might be associated with the need of prolonged postoperative intensive care medicine which in turn might impair intended operative benefit. Therefore, we analyzed prolonged mechanical ventilation (PMV) as an indicator variable for such intensive care treatment with regard to potential correlations with mortality in this vulnerable patient cohort. Between 2012 and 2018, 177 consecutive patients received stabilization surgery for PSD at the authors\u2019 neurosurgical department. PMV was defined as postoperative mechanical ventilation of more than 24 h. A multivariable analysis was performed to identify independent predictors for 30-day mortality. Twenty-three out of 177 patients (13%) with PSD suffered from postoperative PMV. Thirty-day mortality rate was 5%. Multivariable analysis identified \u201cspinal empyema\u201d (The online version contains supplementary material available at 10.1007/s10143-023-02016-1. Pyogenic spondylodiscitis (PSD) represents about 3\u20135% of all forms of osteomyelitis with a continuously increasing incidence up to currently 6.2\u20137.4/100,000 patients per year worldwide . The aimAll consecutive patients with the ICD code diagnosis of discitis or pyogenic vertebral osteomyelitis with infection of the intervertebral disc aged \u2265 18 years who had undergone surgical procedures with instrumentation and/or fusion for PSD at the authors\u2019 neurosurgical department between 2012 and 2018 were entered into a computerized database . Information collected for each patient included sociodemographic characteristics, location of the spinal infectious disease, number of affected spinal levels, and neurological status at admission among others. Indication for an open surgical procedure was given in case of neurological impairment, osseous destruction with spinal instability/deformity or stenosis of spinal canal and compression of neuronal structures, relevant epidural empyema and/or ventral/paravertebral abscess inaccessible for puncture, failure of conservative treatment with progressive disease, or intractable pain.Depending on the site, extent and severity of the infectious spinal disease dorsolateral as well as ventral approaches were performed. In case of severe instability, a decision for a staged procedure (360\u00b0 stabilization) was made. Pathogen detection was defined as positive microbiological cultivation out of preoperatively taken blood cultures and/or out of intraoperatively obtained bone or disc samples with pathogen-specific resistograms. In case of unsuccessful germ cultivation, diagnosis of PSD was made by means of histopathological pathogen detection from intraoperative bone and/or disc samples as well as consistent correlation of radiological signs, systemic inflammatory blood values, and intraoperative macroscopic and microscopic tissue appearance. In case of successful germ detection, a specific antimicrobial therapy was started in accordance to pathogen-specific resistograms. In some patients with septic constellations and/or preexisting antibiotic therapy, empirical and broad antibiotic therapy was necessary. Antibiotics were administered intravenously at least in the first 2 weeks with subsequent potential oralization after C-reactive protein (CRP) dropping below 10 mg/L. A minimum of 6 weeks of antibiotic therapy was standard. Selection of the antimicrobial therapy according to the pathogen spectrum and resistogram occurred in close consultation with the local Institute of Microbiology.PMV was defined as an invasive ventilation period of > 24 h after initial stabilization surgery as previously described . TherefoEarly postoperative complications were assessed by means of a public available list of events introduced by the Agency for Healthcare Research and Quality and the Center for Medicare and Medicaid Services and referred to as patient safety indicators (PSIs) and hospital-acquired conditions (HACs) . PSIs inThe study was conducted in accordance with the Declaration of Helsinki and approved by the Ethics Committee of the University Hospital Bonn . Informed consent was not sought in regard of the retrospective study design.www.graphpad.com). Categorical variables were analyzed in contingency tables using Fisher\u2019s exact test. The Mann-Whitney U test was chosen to compare continuous variables as the data were mostly not normally distributed. Results with p < 0.05 were considered statistically significant. In addition, in order to determine independent predictors of 30-day mortality in patients with surgically treated PSD, a backward stepwise method was used to construct a multivariable analysis using a binary logistic regression.Data analysis was performed using the computer software package SPSS and GraphPad Prism followed by Staphylococcus epidermidis (10%). Clinical admission status revealed PSD-related neurological deficits in 50 of 177 patients (28%). Sixteen patients (9%) died within 30 days following initial stabilization surgery. Twenty-three of 177 patients (13%) received mechanical ventilation of more than 24 h. For further details on patient characteristics, see Table Between January 2012 and December 2018, 177 patients were surgically treated with spinal instrumentation for PSD at our neurosurgical department. Median age at time of surgery was 72 years (IQR 62\u201378 years) including 68 female (38%) and 109 male patients (62%) (Table p = 0.01) as shown in Table p = 0.004). Nine of 23 patients with PMV (39%) suffered from respiratory functional disorder due to preoperatively diagnosed sepsis from pneumonia, urinary tract infection, and/or spinal source of infection including spinal empyema. Four patients (17%) suffered from exacerbated chronic obstructive pulmonary disease (COPD); 2 of 23 patients (9%) developed perioperative or early massive postoperative pulmonary embolism with the need for further mechanical ventilation. Extubation was unsuccessful due to perioperative acute myocardial infarction in 1 patient (4%). One of 23 patients with PMV (4%) suffered from preoperative hemodynamic instability. Nine of 23 patients with PMV (39%) developed early postoperative complications compared to 24 of 154 patients without PMV (16%) (p = 0.02). For detailed information on postoperative complication profiles, see Supplementary Table Thirty-day mortality in patients with postoperative PMV was 39% (9 of 23 patients) compared to 5% (5 of 154 patients) in patients without postoperative PMV 6.2, 95% confidence interval (CI) 1.3\u201330.2), \u201cCCI > 2\u201d , \u201cearly postoperative complications (PSIs)\u201d and \u201cPMV > 24 h\u201d as significant and independent predictors for early postoperative mortality (Nagelkerke\u2019s R2 0.5) , \u201clevel of disease (> 2)\u201d , \u201cpreoperative anticoagulant medication\u201d , and \u201cpreoperative WBC > 12 G/L\u201d as significant and independent predictors for PMV occurrence (Nagelkerke\u2019s R2 0.4) \u201d"} +{"text": "Lung adenocarcinoma (LUAD) patients with elevated breast cancer susceptibility gene 1\u00a0(BRCA1) expression had markedly worse overall survival and progression\u2010free survival compared to those with reduced BRCA1 levels. In contrast, BRCA1 expression did not correlate with survival outcomes in squamous cell carcinoma patients. The overexpression of BRCA1 was an independent risk factor for LUAD and was indicative of an immune\u2010suppressive tumor microenvironment. BRCA1breast cancer susceptibility gene 1DEGsdifferentially expressed genesGOGene OntologyiDCsinduced dendritic cellsLUADlung adenocarcinomaLUSCsquamous cell carcinomaNSCLCnon\u2010small\u2010cell lung cancerOSoverall survivalPFSprogression\u2010free survivalROCreceiver operating characteristicBRCA1) is a tumor suppressor gene that regulates cellular responses to stress through DNA damage repair and is frequently mutated in breast and ovarian cancers [BRCA1 and the relevant poly (ADP\u2010ribose) polymerase inhibitors is related to the prognosis and therapeutic response of other tumors, including lung cancer [BRCA1 for NSCLC have been inconsistent or conflicting, which may be attributed to the small number of patients from a single center or disparity in the ratio of patients with lung adenocarcinoma (LUAD) versus squamous cell carcinoma (LUSC) [BRCA1 in NSCLC and its association with immune infiltration.Non\u2010small\u2010cell lung cancer (NSCLC) is the primary histopathological subtype of lung cancer and accounts for around 82% of all pulmonary neoplasms . Breast g cancer , 4. Howea (LUSC) . The aimBRCA1 expression data of lung cancer patients retrieved from public databases and relevant articles were analyzed. The results demonstrated that BRCA1 RNA and protein levels were significantly upregulated in LUSC and LUAD tissue compared to normal lung tissue or progression\u2010free survival (PFS) in the overall lung cancer cohort, LUAD patients with elevated BRCA1 expression had a markedly worse OS and PFS compared to those with reduced BRCA1 levels. In contrast, BRCA1 expression did not correlate with survival outcomes in LUSC patients \u2010LUAD was integrated and analyzed and exploratory analysis was conducted to fully explore its further application in clinical practice were screened between the BRCA1high and BRCA1low samples, of which 6555 genes were upregulated, and 685 genes were downregulated , whereas an inverse correlation was observed between BRCA1 and Th2 cells ; data curation (lead); formal analysis (lead); investigation ; methodology (lead); project administration (supporting); software ; validation (lead); visualization (lead); writing\u2014original draft (lead); writing\u2014review and editing (supporting). Cong Li: Methodology (supporting); software ; validation (supporting). Shuning Liu: Data curation (supporting); formal analysis (supporting); resources (supporting); software (supporting); visualization (supporting). Zhijun Li: Investigation ; Methodology (supporting); visualization (supporting). Jingtong Zhai: Data curation (supporting); formal analysis (supporting); visualization (supporting). Zhiwu Wang: Conceptualization ; project administration (supporting); supervision (lead); writing\u2014review and editing (lead). Binghe Xu: Conceptualization ; funding acquisition (lead); project administration (lead); supervision (supporting); writing\u2014review and editing (supporting).Cancer Innovation Editorial Board. To minimize bias, he was excluded from all editorial decision\u2010making related to the acceptance of this article for publication. The remaining authors declare no conflict of interest.Professor\u00a0Binghe Xu is the member of the Not applicable.Not applicable.Supporting information.Click here for additional data file."} +{"text": "Diabetic foot osteomyelitis (DFO) is a leading cause of amputations. Rifamycins can penetrate osteoblasts and disrupt biofilm, making them ideal adjunctive antibiotics for DFO. A recent retrospective DFO study found that rifampin adjuvant therapy was associated with significantly higher amputation-free survival rate and randomized clinical trials to establish whether adjunctive rifampin therapy improves DFO outcomes are ongoing. Rifamycin use is complicated by drug-drug interactions (DDIs), primarily due to cytochrome P450 induction. However, rifabutin is a less potent and broad cytochrome P450 inducer versus rifampin. We evaluated DDI predicted rates of rifampin and rifabutin in a DFO cohort.We conducted a retrospective cohort study of all patients hospitalized with DFO between 2017 and 2019 at Grady Memorial Hospital . We queried discharge billing records using ICD-10 codes for DFO and performed a chart review to confirm the diagnosis. We used the Lexicomp DDI tool to determine the number of potential DDIs between all medications prescribed upon discharge and rifampin or rifabutin.S. aureus was identified in 79 (33%). Eighty-nine (27%) patients were prescribed medications with a class X (\u201cavoid combination\u201d) rifampin interaction versus 4 (1%) patients who were prescribed drugs with a rifabutin class X interaction. Two-hundred-and-forty (73%) patients had prescriptions for medications with a class D (\u201cavoid combination\u201d) rifampin interaction versus 15 (5%) with a class D rifabutin interaction. The most common medications with class X rifampin DDIs were proton pump inhibitors (20%) and directly acting oral anticoagulants (5.7%), while those for class D rifampin DDIs were atorvastatin (35%) and clopidogrel (12%).There were a total of 530 hospital admissions among 330 unique patients for DFO of which 70% were male and 80% were Black. The mean hemoglobin A1c was 9.4%. Chronic kidney disease was present in 69% of patients. Among 239 DFO cases with a culture, A high percentage of patients with DFO were prescribed medications with significant rifampin DDIs compared with rifabutin, supporting the need to investigate rifabutin for adjunctive DFO therapy.All Authors: No reported disclosures"} +{"text": "The association of diabetes mellitus, both type 1 (T1DM) and type 2 (T2DM), with increased cardiovascular risk is well known and cardiovascular disease (CVD) events. An analysis from the MESA cohort also found that CAC improved prediction of CHD and CVD more than high CIMT or presence of carotid plaque . For strDespite the consolidation of cardiovascular risk heterogeneity among diabetics, stratified either by a risk score or imaging methods, further investigation should define whether a differential therapeutic approach based on calculated risks or high CIMT/CAC will translate in clinical outcome benefit."} +{"text": "Pneumocystis jirovecii pneumonia (PJP). The clinical utility of BDG, however, is limited because of uncertainty in interpreting quantitative values. A correlation was drawn between quantitative BDG and the diagnosis of PJP in hospitalized patients living with human immunodeficiency virus (HIV) at four affiliate campuses of the Montefiore Health System in the Bronx, NY.-\u03b2-D-glucan (BDG) has been utilized as an indirect marker for Pneumocystis direct fluorescent assays (DFA), polymerase chain reactions (PCR) either from bronchoalveolar lavage (BAL) fluid or sputum, and quantitative BDG assays either from serum or BAL fluid were retrieved from the Montefiore microbiology laboratory (date range: 8/1/18 - 8/31/22). Results from hospitalized patients with HIV whose CD4 count within 3 months were available, with either PJP DFA or PCR result within 7 days of serum BDG and lactate dehydrogenase (LDH) test were selected (n = 146). CD4 count was grouped into < 50 (n = 106), 50-100 (n = 15), 100-200 (n = 16), and >200 (n = 9). BDG, LDH, and CD4 were considered as input variables in a logistic regression model for true PJP status. The area under the curve (AUC) of the empirical receiver operating characteristic (ROC) curve with 95% confidence intervals (CI) was estimated based on trapezoidal area. Comparisons between ROC curve areas were performed via contrast matrix to take differences of the AUC of the empirical ROC curves and chi-square testing.All p = 0.025). Adding CD4 group did not influence AUC.Of the 146, 29 had PJP detected and 117 did not. When BDG only was considered, the AUC of the ROC curve was 0.78. When LDH quartiles were considered with BDG, there was a statistically significant increase in AUC improved when LDH quartiles were used in conjunction with BDG, suggesting a better prediction of PJP diagnosis when BDG and LDH were considered together. This result supports further investigation of clinical markers of PJP given the limitations of establishing a microbiologic diagnosis. Further steps of the investigation will involve analyzing if other clinical factors including HIV viral load, oxygen saturation, and radiographical reading, influence predictability of PJP.All Authors: No reported disclosures"} +{"text": "In understanding clonal evolution in cancer, whole genome sequencing (WGS) provides unparalleled data that can be leveraged to derive tumor phylogenies. Specifically, estimates of the cancer cell fraction (CCF), that is the fraction of tumor cells bearing a set of somatic mutations, is highly informative by defining which mutations are clonal and are associated with the most recent common ancestor (MRCA) of all tumor cells, and which mutations occur subclonally as later events. By clustering all observed somatic mutations based on their CCF and subsequent phylogenetic tree inference, derived tumor phylogenies provide an even more detailed view of the tumor history than mere single point estimates of CCF of driver variants, as they determine the relative order of all events and infer evolutionary trajectories. These evolutionary trajectories inform which driver genes are early events in tumorigenesis, that dysregulate cellular molecular pathways critical to clonal proliferation.BRAFV600E mutation, making it a rare monogenic cancer. A large number of additional exonic mutations have been observed in HCLc, but none are pervasive across all tumors . BRA. BRA2]. BCL6) and ZFP36 ring finger protein like 1 (ZFP36L1) were recurrently mutated in 50% of HCLc cases and prioritized by all noncoding scoring algorithms of a HCLc cohort, suggesting a potential function in the pathogenesis of this disease [Chromosomal instability (CIN) is a hallmark of malignant transformation with copy number aberration (CNA) being ubiquitous across tumor types. Analysis of CNA showed a relatively stable genome in the HCLc cases (ploidy \u22482). We observed one large CNA (chr5 gain) and two focal CNAs (chr17q25.1 gain and chr20q11.22 LOH) Fig.\u00a0. Althoug disease . RecurreTRB) locus COSMIC signatures SBS1, SBS5 and SBS9 in our HCLc cohort with similar relative contributions and assigned the driver variants to their respective clusters Fig.\u00a0. In all lly Fig.\u00a0. All sublly Fig.\u00a0. InteresBRAFV600E alone may suffice to drive expansion of the ancestral clone (MRCA) to achieve malignant transformation, in at least a subset of HCLc. In separate pathways, BRAFV600E may also require additional mutational drivers, that differ between tumors, for the MRCA to out-compete other cells and become clonal. These findings reveal heterogeneous genetic origins and subsequent subclonal diversification in HCLc disease.Our findings conclude that Supplementary Data"} +{"text": "Correction to: Mycorrhiza (2023) 33:187\u2013197 https://doi.org/10.1007/s00572-023-01112-wThe first Fig.\u00a01 citation should have been Fig.\u00a01aThe text should be \"occurrence\" instead of \"relative\"The text should be \"subsequent\" instead of \"community\"The text should be \"alpha\" instead of \"beta\"The text should be \"80% and 60% occurrence\" instead of \"41% and 42% relative\"The text should be \"frequent\" instead of \"dominant\"The text should be \"an occurrence frequency of 50%, followed by Tomentella sp. 1 (46%)\" instead of \"a relative frequency of 31%, followed by Tomentella sp. 1 (28%)\"The text below under Result section should have been corrected as follows:The text should be \"age\" instead of \"growth\"The text should be \"dispersal\" instead of \"migration\"The text \"indirectly\" should be deletedThe text below under Discussion section should have been corrected as followsThe original published version of the above article contained errors. The following data should have been corrected as follows:The original article has been corrected."} +{"text": "Blood Cancer Journal 10.1038/s41408-023-00794-x, published online 06 March 2023Correction to: Following the publication of this article the authors noted and error in Figure 4, \u201cKaplan Meier curves for time to next treatment (TTNT) according to sequential MRD measurements\u201d. The figure should have displayed only patients who had MRD taken at least 1 year apart (as defined by IMWG), but instead it incorrectly included an additional 8 patients with sequential measurements taken only at least 6 months apart.The correct figure shows results based on the first MRD taken at least 1 year apart following day 100 evaluation, and should have appeared as below:The original article has been corrected."} +{"text": "Although incidence of Clostridioides difficile infection (CDI) historically is higher in white compared to black patients, the role of race or social determinates of health on rCDI is unclear. This study explores social determinants of risk for rCDI and treatment with FMT.Recurrent CDI (rCDI) can be deadly and live biotherapies such as fecal microbiota transplantation (FMT) are now recommended at 2Georgia\u2019s Emerging Infections Program (EIP) (supported by CDC) conducts CDI surveillance in 8 metropolitan Atlanta counties. Incident CDI (+toxin or molecular assay) in resident of catchment area (without previous + within 14 days) between 2016 and 2019 were categorized as single episode or rCDI ( >1 episode within 365 days) and geocoded (ArcGIS)-linked to CDC\u2019s 2020 Social Vulnerability Index (SVI) data. Numerical values of each SVI variable (Table 1) were assigned to each patient. SVI theme and composite SVI were assessed as predictors for each rCDI and FMT receipt with most vulnerable defined categorically (top quartile) in log binomial logistic regression. Spatial analysis produced spot map of FMT, and choropleth maps of the CDI and rCDI rates per 100,000 persons.Among the 13,835 residents with CDI, 3,038 (22.0%) had rCDI and 250 (1.8%) had an FMT. Housing type and transportation was the only SVI predictor for rCDI while it was one of several SVI themes predictive of not receiving FMT (Table 2). Adjusting for age and sex, the most vulnerable patients in housing and transportation theme were 13% more likely to have recurrence (Table 3). Independent predictors of FMT receipt included younger age and female sex. Importantly, patients living in a largely minority census tract were 37% less likely to receive an FMT and those in a largely uninsured tract 39% less likely (Table 3). Spatial analysis illustrates a strong discordance between FMT receipt and rCDI rates .More socially vulnerable populations, defined by access to transportation and the level of household crowding, are more likely to develop rCDI. Likelihood of receiving FMT is substantially reduced among uninsured and persons of black race; both associations need consideration as novel live biotherapeutic products become available.Nitin Gupta, MD, Abbvie: Advisor/Consultant|Amgen: Grant/Research Support|Bristol Myers Squibb: Advisor/Consultant|Bristol Myers Squibb: Grant/Research Support|Janssen: Advisor/Consultant|Janssen: Grant/Research Support|Pfizer/Arena: Advisor/Consultant|Pfizer/Arena: Grant/Research Support"} +{"text": "Transcatheter tricuspid valve repair (TTVR) is a promising therapy for severe tricuspid regurgitation (TR) that requires accurate navigation within the right heart cavities [cavities . We hereA patient-specific 3D heart model is established based on a 3D full-volume echocardiographic image that is overlaid on fluoroscopy . The heart model visualizes the echocardiography-derived anatomical cavities, and the fluoroscopy-derived device merged into a single display. In addition, customized anatomic landmarks are set at the anterior and posterior sites of the tricuspid valve in TOE imaging and superimposed on fluoroscopy.The integrated echo-derived right heart cavities on fluoroscopy facilitates device navigation from the vena cava into the right atrium in the direction of the tricuspid annulus without getting stuck at the interatrial septum or right atrial appendage or causing tissue injury Fig.\u00a0B. Anatomreal-time fusion imaging to simplify TTVR. Whether fusion imaging guided TTVR improves safety and reduces the procedure time and radiation exposure requires investigation in future trials.Here we demonstrate how to use Supplementary file1 (MP4 1993 KB) Video 1: Transesophageal 3D-echocardiographic images of the right heart cavities (left panel). Real-time fusion imaging assisted navigation of the TTVR device through the right atrium pointing to the tricuspid annulus (right panel)Supplementary file2 (MP4 1905 KB) Video 2: Biplane tranesophageal echocardiographic images after safe device passage into the right ventricle (left panel). Fusion of echocardiographic and X-ray images provides the superimposed echo-derived heart cavities and anatomical landmarks onto live X-ray fluoroscopy (right panel)Supplementary file3 (MP4 1119 KB) Video 3: Biplane transesophageal echocardiographic images showing successful grasping of the septal and posterior leaflet of the tricuspid valveSupplementary file4 (MP4 1023 KB) Video 4: Transgastric echocardiographic image showing successful grasping of the septal and posterior leaflet of the tricuspid valveSupplementary file5 (MP4 1458 KB) Video 5: Biplane transesophageal echocardiographic images after leaflet grasping (left panel). Fusion of echocardiographic and X-ray images provides the superimposed echo-derived heart cavities and anatomical landmarks onto live X-ray fluoroscopy (right panel)Below is the link to the electronic supplementary material."} +{"text": "Dear Editor,\u22124) in peripheral blood (PB) at end of treatment than those receiving BR, regardless of presence of high-risk biomarkers [\u22122). Moreover, the median time from MRD conversion to clinical progression was 25.2\u00a0months, suggesting that early intervention before clinical progression in patients with MRD conversion might be a clinical option that merits testing in clinical trials. Fixed-duration therapy using venetoclax is being introduced for first-line treatment of CLL. Consequently, monitoring of MRD is increasingly important for CLL management, but reports on MRD monitoring in these patients remain rare in Japan. Here, we report MRD monitoring results of our phase 1/2 study of venetoclax with/without rituximab in patients with rrCLL/small lymphocytic lymphoma (SLL) [Monitoring of measurable residual disease (MRD) can assess depth of response and predict prognosis in patients with chronic lymphocytic leukemia (CLL) treated with immuno-chemotherapy or targeted therapies such as venetoclax monotherapy or venetoclax plus anti-CD20 monoclonal antibodies, which effectively eradicate CLL cells. In the phase 3 MURANO trial comparing venetoclax plus rituximab (VenR) against bendamustine plus rituximab (BR) in patients with relapsed or refractory CLL (rrCLL), patients in the VenR arm had better progression-free survival (PFS) and a higher rate of undetectable MRD . MRD data were available for two patients (Patients 1 and 2) in Arm B and four patients (Patients 3\u20136) in Arm D. However, PB samples were unavailable for the other six patients due to SLL in two patients, discontinuation of the study because of adverse events and progressive disease in two patients, and unreported reasons in the remaining patients. We evaluated achievement of uMRD or MRD4 as defined by\u2009<\u20091\u2009\u00d7\u200910\u22124 MRD [Samples for MRD measurements were prospectively obtained from PB at baseline (both arms), day 1 of week 24 (Arm B), day 1 of week 30 and every 12\u00a0weeks thereafter (Arm D), and within 3\u00a0months after complete remission (CR), after CR with incomplete blood count recovery, or when radiographic partial response (PR) criteria were first met (both arms). MRD was measured using the clonoSEQTumor responses and MRD are presented in Table \u00ae assay is feasible in Japanese patients with rrCLL who received venetoclax. uMRD was achieved in all patients treated with VenR with available MRD data.In conclusion, MRD monitoring using the clonoSEQ"} +{"text": "The suppression of immune system in transplant patients increases the risk of Cytomegalovirus reactivation, which leads to higher rates of morbidity and mortality. Many risk factors have been identified to be associated with increased risk of CMV reactivation including mismatched human leukocyte antigen (HLA) transplant and myeloablative conditioning.All patients =/ > 14 years that underwent allo-SCT from April 2014 to April 2022 with complete medical records were included. Data collected include CMV serostatus, CMV viral load, time to CMV reactivation, conditioning regimen, use of in-vivo T-cell depletion for graft vs. host disease (GvHD)prophylaxis, and GvHD prophylaxis medications if usedWe included 404 patients with an average age of 29 years and 42% of female sex. Patients were divided into two different groups based on underlying disease for which All-SCT was indicated.183 patients (Group 1) underwent Allo-SCT for underlying hematological malignancy mainly acute myeloid leukemia (N= 64) and acute lymphoblastic leukemia (N=73) and remaining patients distributed between CML, Hodgkin and non- Hodgkin lymphomas. Others (N = 221) had sickle cell anemia and a plastic anemia. More than 75% of patients received myeloablative conditioning regimen in Group1 compared to Group2 were most patients received non-myeloablative regimen. Most patients received matched \u2013 related human leukocyte antigen (HLA). CMV seropositive recipients constitute 95% of all patients; 89% of patients experienced CMV reactivation within 13 days post SCT and had an average initial viral load of 51 IU during reactivation. 220 patients (62%) were able to clear CMV spontaneously within a median of 50 days post reactivation. 90 day non-relapse mortality and 1 years overall survival were 2.7% and versus 1.4% and 80% versus 94% in Group 1 and Group 2 respectively.Our population is considered at higher risk of CMV reactivation due to the high prevalence of CMV seropositivity. Identifying factors that could contribute to CMV reactivation would help in identifying patients at highest risk and therefore, identify the best approach to manage CMV reactivation in those patients.All Authors: No reported disclosures"} +{"text": "As the pathogenesis of vitiligo remains unknown, the ineffective rate of various treatments for vitiligo patients has reached 50% . TherefoWe have obtained McSCs in a functional state using a PCT-protected technical method and implanted them and melanoblasts to an area under the epidermis. Continuously activated by a 308-nm excimer laser in vitro, McSCs in the ORS were transformed into mature MCs and migrated along the ORS to multiple hair follicle orifices in the vitiligo area or sebaceous gland openings in the hairless area to achieve central-type repigmentation with no color difference. McSC transplantation addresses the issue of MC sources for patients with vitiligo and provA 35-year-old male patient presented with multiple depigmented leukoplakia on the right side of the face for 17\u00a0years Figs. .Fig. 1FiSupplementary file1 (PDF 2373 KB)Supplementary file2 (PDF 51673 KB)Supplementary file3 (PDF 855 KB)Supplementary file4 (PDF 2109 KB)Supplementary file5 (PDF 1563 KB)Below is the link to the electronic supplementary material."} +{"text": "Approximately 4.4 million children die peripartum annually, primarily in low- and middle-income countries. Accurate mortality tracking is essential to prioritising prevention efforts but is undermined by misclassification between stillbirths (SBs) and early neonatal deaths (ENNDs) in household surveys, which serve as key data sources. We explored and quantified associations between peripartum provider-mother interactions and misclassification of SBs and ENNDs in Guinea-Bissau.Using a case-control design, we followed up on women who had reported a SB or ENND in a retrospective household survey nested in the Bandim Health Project\u2019s Health and Demographic Surveillance Systems (HDSS). Using prospective HDSS registration as the reference standard, we linked the survey-reported deaths to the corresponding HDSS records and cross-tabulated SB/ENND classification to identify cases (discordant classification between survey and HDSS) and controls (concordant classification). We further interviewed cases and controls on peripartum provider-mother interactions and analysed data using descriptive statistics and logistic regressions.We interviewed 278 women (cases: 63 (23%); controls: 215 (77%)). Most cases were SBs misclassified as ENNDs (n/N\u2009=\u200949/63 (78%)). Three-fourths of the interviewed women reported having received no updates on the progress of labour and baby\u2019s health intrapartum, and less than one-fourth inquired about this information. In comparison with births where women did inquire for information, misclassification was less likely when women did not inquire and recalled no doubts about progress of labour (odds ratio (OR)\u2009=\u20090.51; 95% confidence interval (CI)\u2009=\u20090.28-0.91), or baby\u2019s health . Most women reported that service providers\u2019 death notifications lasted <5 minutes (cases: 23/27 (85%); controls: 61/71 (86%)), and most often encompassed neither events leading to the death (cases: 19/27 (70%); controls: 55/71 (77%)) nor causes of death (cases: 20/27 (74%); controls: 54/71 (76%)). Misclassification was more likely if communication lasted <1 compared to 1-4 minutes and if a formal service provider had informed the mother of the death compared to a family member .Peripartum provider-mother interactions are limited in Guinea-Bissau and associated with birth outcome misclassifications in retrospective household surveys. In our study population, misclassification led to overestimated neonatal mortality. An estimated two million children are stillborn and 2.4 million die during their first month of life globally each year , 97% of Accurate and timely mortality tracking is essential for prioritising mortality prevention efforts and monitoring progress towards the global mortality targets -9. Howev+) [Such surveys have most commonly used the full birth history (FBH) approach to capture mortality. In FBHs, women are interviewed on their history of live births, child survival status, and child age at death, thereby omitting adverse non-live pregnancy outcomes including stillbirths . To capt+) . More re+) ,16. Sinc+) ,17.+ survey data with that from a prospectively followed health and demographic surveillance system (HDSS) cohort in urban Guinea-Bissau, the FBH+ data overestimated neonatal mortality by 5-20% [+ data, and 63% of deaths reported in FBH+, but not in HDSS data [There are several data quality challenges associated with survey-derived stillbirth and neonatal mortality estimates ,18,19, oAccurate classification of stillbirths and neonatal deaths based on household surveys presupposes maternal knowledge of the child\u2019s vital status at birth. Several determinants of maternal misclassification have been discussed, including compromised recall and numeracy , social + or FPH modules (EN-INDEPTH study) conducted at the Bandim Health Project (BHP) in 2017-2018 [We applied an unmatched case-control design, defining cases and controls based on discordant (cases) and concordant (controls) maternal reporting of a child death as stillbirth or early neonatal death (i.e. death during the first week of life) between two data sources: a retrospective population survey mimicking DHS data collection using standard FBH017-2018 , and pro017-2018 ,31. To iWe recruited participants in 2021-2022 from respondents of the EN-INDEPTH study, which collected retrospective self-reported information on stillbirths and early neonatal deaths among women in BHP\u2019s urban and rural HDSS with a registered birth outcome during the last five years ,32. BHP\u2019We invited eligible women residing in the HDSS area to participate in a household interview (case-control interview). They were initially contacted by the HDSS staff routinely collecting household information and asked if they were willing to be visited by a colleague collecting more information on women\u2019s experiences of losing a child. Provided acceptance, one of two specially trained female HDSS field workers visited the women for the case-control interviews. Following a consent process in all statistical tests (+ and FPH) , this was the case for only 67% of the HDSS-classified stillbirths (n/N\u2009=\u200999/148) ), and close to half of the women were aged 20-29 years at the time of the EN-INDEPTH interview (n\u2009=\u2009132 (47%)). The overall level of educational attainment was low, with 35% of the women never having attended school (n\u2009=\u200996) and 28% having attended only primary school (n\u2009=\u200979). Three-quarter of the women had at least one pregnancy prior to the one interviewed about (n\u2009=\u2009207 (74%)), and 29% of those women had experienced a previous perinatal death (n\u2009=\u200960). Based on wealth quintiles, women from the urban study area appeared poorer than those from the rural areas. Fula/Mandinga were the biggest ethnic group (n\u2009=\u200982 (29%)), followed by Pepel (n\u2009=\u200966 (24%)) (P\u2009=\u20090.015) with more controls than cases having been informed about serious dangers to the baby\u2019s health (controls: n\u2009=\u200921 (12%); cases: n\u2009=\u20093 (5%)) ) and had been assisted by a skilled birth attendant (n\u2009=\u2009225 (81%)). Among the women giving birth in a health facility, one in five had been referred (n\u2009=\u200949 (21%)). Most women had given birth vaginally (n\u2009=\u2009221 (79%)), 15% had a C-section (n\u2009=\u200943). One in five women reported labour of >18 hours (n\u2009=\u200959 (21%)), and more than half reported intrapartum complications (any: n\u2009=\u2009115 (41%); >1: n\u2009=\u200930 (11%)). Nine per cent of the women had a multiple gestation (n\u2009=\u200924). Most perinatal deaths occurred at the health facility of delivery (n\u2009=\u2009184 (66%)), 21% at home (n\u2009=\u200957), and 9% at another health facility (n\u2009=\u200925) (P\u2009=\u20090.053). Proxy reporting of HDSS data was more common among cases than controls (P\u2009=\u20090.044). The distribution of women who had been interviewed using FBH+ and FPH surveys in the EN-INDEPTH interviews was even and similarly distributed across cases and controls \u2009=\u20090.1-1.2), EN-INDEPTH interviews were conducted at a median of 26 months (IQR\u2009=\u20099-43) and case-control interviews at a median of 74 months (IQR\u2009=\u200959-92). While the distribution of women interviewed above the respective median recall was similar across cases and controls for EN-INDEPTH interviews and case-control interviews, cases tended to have a longer recall length in HDSS interviews ; controls: 31 (14%); baby\u2019s health \u2013 cases: 13 (21%); controls: 31 (14%)). In comparison with deaths where the mother inquired information about the progress of labour and the baby\u2019s health, misclassification was less likely when the mother recalled no doubts . Most women saw, heard, or felt their baby postpartum (cases: 41 (65%); controls: 157 (73%)) and misclassification tended to be more likely when the mother did not see, hear, or feel the baby ; controls: 149 (69%)), 17% of cases (n\u2009=\u200911) and 23% of controls (n\u2009=\u200950) learned about the death indirectly, e.g. by seeing the dead child or overhearing a conversation between service providers and family members. Misclassification tended to be less likely when the woman was not directly informed . Among women who were directly informed, half were informed by a formal service provider (cases: 24 (53%); controls: 67 (45%)), and misclassification was more likely if a formal service provider had informed in comparison with a family member . Half of the women reported that family was present during the death notification. Misclassification was more likely amongst women who had met the informing provider once before compared with women who did not know the provider , but not among women who had met the provider more often. Misclassification was less likely among mothers who learned about the death at the health facility outside the delivery room and maternity ward compared with in the delivery room (Most women reported that service providers\u2019 death notifications were short and lasted <5 minutes (cases: 23/27 (85%); controls: 61/71 (86%)). Misclassification was more likely if the communication was <1 minute compared to 1-4 minutes . The majority of both cases and controls reported that the service provider communicated the death in a way they understood immediately (cases: 23/27 (85%); controls: 60/71 (85%)), told everything they wanted to know (cases: 21/27 (78%); controls: 55/71 (77%)), and paid full attention to them (cases: 24/27 (89%); controls: 60/72 (83%)). Yet most women also reported that the service providers neither mentioned events leading to the death (cases: 19/27 (70%); controls: 55/71 (77%)) nor causes of death (cases: 20/27 (74%); controls: 54/71 (76%)) , and proxy reporting were associated with higher odds of misclassification ). Hence, misclassification was not evenly bidirectionally distributed. This is a similar result to a previous study from urban Guinea-Bissau which also found an overestimation of neonatal deaths in a retrospective household survey . Hence, %). HenceThis study built on data collected during the retrospective EN-INDEPTH population survey mimicking DHS data collection approaches and prosWhile the accurate classification of perinatal deaths as stillbirths or early neonatal deaths is fundamental to target and monitor preventive action, little is known about mechanisms determining maternal misclassification in household surveys. In this study, we found limited peripartum provider-mother interactions during births with adverse outcomes in Guinea-Bissau, which likely compromise maternal knowledge of circumstances surrounding the child death and reporting validity. We also found modifiable factors associated with maternal misclassification including maternal doubts during birth and length and source of child death notifications. Moreover, our findings indicate that misclassification between stillbirths and early neonatal deaths do not cancel each other out but may rather lead to overestimation of neonatal mortality in household surveys, thereby highlighting the need for further research aiming at improving the understanding of determinants of misclassification."} +{"text": "Escherichia coli (APEC) and Enterococcus faecalis in poultry with colisepticemia have become increasingly recognized. Here, we report draft genome sequences of 18 APEC and 18 E. faecalis strains coisolated from lesions of diseased poultry.Coinfections by avian pathogenic Escherichia coli (APEC) and Enterococcus faecalis cause septicemic disease in poultry resulting in substantial mortality, economic burdens, and public health risks from lesions identified grossly as omphalitis or splenomegaly. Samples were incubated aerobically at 37\u00b0C on Trypticase soy agar with 5% sheep blood for 18 to 24\u2009h, and isolates from cultures that contained a mixed growth of only Escherichia coli and Enterococcus were pursued. Species were confirmed by either matrix-assisted laser desorption ionization\u2013time of flight (MALDI-TOF) mass spectrometry or Biolog GenIII microplates using standard methods as described previously was used for APEC DNA isolation, and the MasterPure Gram-positive DNA purification kit was used for E. faecalis DNA isolation. DNA quality and quantity were assessed with a Qubit 4.0 fluorometer and the double-stranded (dsDNA) high-sensitivity assay kit . Pooled libraries with an insert size of ~350\u2009bp were prepared with the Nextera DNA flex library prep kit . Sequencing was completed on the Illumina MiSeq platform with paired-end 150-nucleotide reads and V2 chemistry using a MiSeq v2 reagent kit (500 cycles) according to the manufacturer\u2019s instructions. Sequencing reads were assessed for quality using the MicroRunQC version 3 workflow within GalaxyTrakr (de novo and arranged into scaffolds in CLC Genomics Workbench version 22.0.1 (Qiagen). All sequences were annotated using the National Center for Biotechnology Information (NCBI) Prokaryotic Genome Automatic Annotation Pipeline version 6.3 (PGAAP) for APEC and 56,782 to 1,499,542\u2009bp for E. faecalis. Isolate details, sequencing and assembly statistics, and GenBank accession numbers for raw reads and draft genome assemblies are presented in APEC and PRJNA293225 (Escherichia coli) and PRJNA837978 . The versions described in this announcement are the second versions. Raw read files and genome assemblies can be accessed with the Sequence Read Archive (SRA) and genome accession numbers in The draft genome assemblies have been deposited in GenBank under BioProject accession numbers"} +{"text": "Mycobacterium tuberculosis bacteria whose essential gene information wasdownloaded from ePath and NCBI database for mapping and matching essential genes by using a genome extraction program. The selectionof key features was performed by using Genetic Algorithm. For each of three classifiers, 80%, 10% and 10% of subset key features wereused for training, validation and testing, respectively. Experimental results (10-f-cv) illustrated that DNN (proposed), DT, and SVMachieved AUC of 0.98, 0.88 and 0.82, respectively. DNN (proposed) outperformed DT and SVM. The higher prediction accuracy ofclassifiers was observed because of using only key features which also justified better generalizability of classifiers and efficiencyof key features related to gene essentiality. Besides, DNN (proposed) also showed best prediction performance while compared with otherpredictors used in previous studies. The genome extraction program was developed for mapping and matching of essential genes betweenePath and NCBI database.Accurate investigation and prediction of essential genes from bacterial genome is very important as it might be explored ineffective targets for antimicrobial drugs and understanding biological mechanism of a cell. A subset of key features data obtainedfrom 14 genome sequence-based features of 20 strains of Mycobacterium tuberculosis strains has shown higher genetic diversity correspondingto patterns of human migration, which suggests the co-evolution of distinct lineage with various human populations11][11]Myco[11]Mycoulations. Very reulations used keyulations used SVMulations used anoulations used 13 ulations reportedulations used amulations and otheulations. In thisThe information about essential genes (Gene locus IDs) of 20 strains of M. tuberculosis bacteria was taken from e-Path hypotheticalessential gene database . The genThe results (Table 1 - see PDF) illustrates the details of essential and non-essential genes of M. tuberculosis strains and it wasseen that essential and non-essential gene ratio stands 1:2.9 which indicated an imbalanced dataset. Thus with a view to enhanceprediction performance of three classifiers, strategies for down-sampling and redundancy reduction in non-essential genes(majorityclass) were applied as imbalance datasets creates problems for classifiers . AccordiIn respect of each gene in the balanced dataset, 14 sequence based features including amino acid length and codon frequencies wereextracted using CodonW , a multiThe generalization ability and accuracy performance of prediction models are straight way related to selection of key features13]16][16]13][[16]13]Considering the prediction of essential genes as a binary classification, three Machine Learning classifiers namely Lib SVM - RBF,C 4.5 DT and a newly designed DNN with MLP were evaluated on subset of key features and the findings showed no logical contradictionwith previous works 11]16][16]11][16]11]Relu = f(x) = max (1)I = Exp(Zi)/\u03a3i=1to K Exp(Zi) (2)Softmax(z)i represents the ith element of input to the softmax function.Here, Zn(Sensitivity),Sp(Specificity), PPV, Accuracy(Acc), NPV though AUC score is considered as primary evaluation measure for classifiersperformance. The other performance measures are appended below:The classifiers were trained with 80% of total subset key features and 10% data was used for validation. The testing of classifierswas executed with remaining 10% of subset key features. The entire training sub dataset were divided into 10 equal divisions and10-f-cv was performed with three classifiers by using subset key features. The accuracy metrics were calculated from the confusionmatrix computed for each of ten divisions of dataset during training and the mean of the accuracy metrics was calculated to determinefinal accuracy metrics after 10-fold cross validation (10-f-cv). Area under Receiver operating characteristics(AU-ROC) curves weregenerated with three classifiers using subset key features and results showed the numerical score of AUC, SSensitivity = True Positives/True Positives + False Negatives (3)Specificity = True Negatives/ False Positives + True Negatives (4)Positive Predictive Value (PPV/Precision) = True Positives/True Positives + False PositivesNegative Predictive Value(NPV)=True Negatives/True Negatives + False Negatives (6)Accuracy = TruePositives+TrueNegatives/True Positives+False Negatives+TrueNegatives+False Positives (7)In this study, three classification algorithms - DT, SVM and DNN were explored with a view to evaluate the performance of eachclassifier with key features and analysed the impact of key features selection on the accuracy of classifiers. It was observed fromTable 2(see PDF) that DNN was highly sensitive to key feature selection, where SVM and DT showed less sensitivity in accuracy withkey features. However, DNN outperformed SVM and DT in respect of AUC. The average AUC scores of three predictors (Table 2-see PDF) didnot contain any logical contradiction to previous work on prediction of essential genes of other bacteria11]14][14]11][[14]11]In this study, SVM, DT and a newly designed DNN with MLP approach were explored and evaluated on genome sequence based key featureswhich were screened by using genetic algorithm based on Random Forest classifier. The DNN model (proposed) with highest predictionaccuracy outperformed SVM and DT. Therefore, DNN model can be a valuable classifier for prediction of essential genes as potentialdrug targets. The results of the study justified the better generalizability of classifiers and effect of selected features onpredictors' accuracy. There is an ample scope for further research work on the improvement of generalization ability of classifier byfine-tuning the discriminatory features."} +{"text": "Otaria flavescens) in Peru that seemed to be associated with highly pathogenic avian influenza A(H5N1) virus infection. The transmission pathway may have been through the close contact of sea lions with infected wild birds. We recommend evaluating potential virus transmission among sea lions.We report a massive mortality of 5,224 sea lions ( Sula variegata), guanay cormorants (Leucocarbo bougainvilliorum), and Peruvian pelicans (Pelecanus thagus) (Otaria flavescens) on the coast of Peru within a few months; the sea lions manifested neurologic and respiratory signs. Clinical signs we observed suggest they were affected by HPAI H5N1, which was later confirmed by government and scientific reports (The panzootic (2020\u20132023) caused by the highly pathogenic avian influenza (HPAI) A(H5N1) caused numerous global outbreaks in 2022 virus, Peru."} +{"text": "Inhalation injury is associated with increased incidence of pneumonia. Bacteria are commonly isolated from bronchoalveolar lavage (BAL) within 12-48 hours of injury with unclear clinical significance. We aimed to determine if grade of inhalation injury is associated with different bacterial species isolated from initial BAL, and if higher inhalation injury grade is associated with increased risk of pneumonia.Abbreviated injury scores (AIS) and clinical microbiology isolates from diagnostic bronchoscopy with BAL performed within 48 hours of injury among adults with inhalation injuries from 2009-2022 were extracted from medical records at an ABA-verified burn center. CDC PNU1 criteria was used to identify pneumonia >48 hours after admission. Modified Poisson regression with robust standard errors was used to assess the association of inhalation injury grade and subsequent pneumonia.Two hundred forty-six patients with inhalation injury who underwent diagnostic bronchoscopy for airway inspection and surveillance BAL within 48 hours of injury were included in this analysis. Fourteen (5.7%) were grade 0, 106 (43.1%) grade 1, 75 (30.5%) grade 2, 51 (20.7%) grade 3-4. The most common bacterial isolate was Streptococcus spp irrespective of AIS grade. Higher AIS grade was associated with increased incidence of pneumonia after adjustment for age, sex, and total body surface area of burn involvement.Bacterial species isolated did not differ by inhalation injury grade. Higher inhalation injury grade was associated with increased risk of pneumonia.Providers should be vigilant that patients with higher grade of inhalation injury are at increased risk of pneumonia. Further research is urgently needed to assess the clinical significance of organisms isolated on initial BAL and their role \u2013 if any \u2013 on the development of pneumonia."} +{"text": "The retraction has been agreed following an investigation into concerns raised by a third party, which revealed inappropriate duplications between this and other published articles [The above article, published online on 15 September 2018 in Wiley Online Library (articles , 4. Thus"} +{"text": "The Centers for Medicare Medicaid Services (CMS) Plan Quality and Performance Program, or Star Ratings Program, allows Medicare beneficiaries to compare quality of care among available Medicare Advantage prescription drug (MA-PD) plans and stand-alone prescription drug plans (PDPs). Health plans have increased intervention efforts and applied existing care management infrastructure as an approach to improving member medication adherence and subsequent Part D star rating performance. Independent Care Health Plan (iCare), an MA-PD plan; MedImpact Healthcare Systems, Inc. (MedImpact), a pharmacy benefits manager; and US MED, a mail order pharmacy, partnered to engage and enroll iCare\u2019s dual-eligible special needs population in an intervention designed to improve patient medication adherence and health plan performance for 3 Part D patient safety outcome measures: Medication Adherence for Oral Diabetes Medications (ODM), Medication Adherence for Hypertension (HTN), and Medication Adherence for Cholesterol (CHOL).To (a) assess the effectiveness of a coordinated member-directed medication adherence intervention and (b) determine the overall impact of the intervention on adherence rates and CMS Part D star rating adherence measures.Administrative pharmacy claims and health plan eligibility data from MedImpact\u2019s databases were used to identify members using 3 target medication classes. Adherence was estimated by the proportion of days covered (PDC) for all members. Those members considered at high risk for nonadherence were prioritized for care management services. Risk factors were based on members\u2019 use of more than 1 target medication class, newly started therapy, and suboptimal adherence (PDC\u2009 less than \u200980%) in the most recent 6-month period. Data files listing member adherence rates and contact information were formatted and loaded monthly into iCare\u2019s care management system, which triggered an alert for care coordinators to counsel members on the importance of adherence and offer the members an option for monthly 30-day supply medication delivery via US MED. Member adherence rates were calculated 9 months pre- and postimplementation for all members and adjusted by length of member enrollment based on CMS technical specifications. Regression analysis assessed pre-post changes in rates comparing 2 intervention groups: (1) members receiving iCare counseling only (iCare-only) and (2) members receiving counseling and medication delivery (iCare\u2009+\u2009US MED). To evaluate the overall impact of the intervention, iCare\u2019s adherence rates and iCare\u2019s measure-specific star ratings for the 2011 and 2012 calendar years (CMS measurement years) were compared with the national MA-PD plan contract average and with a health plan similar in member characteristics but without adherence intervention exposure.A total of 2,700 members were initially targeted for referral to iCare care management and US MED customer service specialist teams. Between April 2012 (implementation date) and January 2013, 1,302 (48.2%) members enrolled in the US MED component of the intervention. Seventy-six percent of identified members were nonadherent (PDC\u2009 less than \u200980%) to 1 of the 3 target medication classes, and 32% of members were nonadherent to more than 1 target medication class. Pre-post absolute average adherence rates increased for the iCare-only group and the iCare-US MED group . From 2011 to 2012, iCare adherence rates increased by absolute differences of 15.2, 9.2, and 10.1 percentage points for ODM, HTN, and CHOL measures, respectively, compared with the average MA-PD plan contract differences and the comparator health plan differences . Increases in iCare\u2019s adherence rates were associated with significant increases in iCare\u2019s 2014 adherence measure star ratings , which contributed to increases in the Drug Plan Quality Improvement measure (2 stars to 4 stars) and iCare\u2019s overall Part D star rating (3 to 3.5 stars).Members in this MA-PD plan dual-eligible population benefited from multiple points of contact to achieve increased adherence. Health plans can use network pharmacies, care management staff, and their pharmacy benefits managers to collaborate and implement interventions aimed to improve members\u2019 adherence to targeted maintenance medications and overall health plan quality performance and star ratings."} +{"text": "This study investigated the effects of prematurity and ROP on visual acuity and VRQoL in adults (18\u201352 years).The Gutenberg Prematurity Eye Study is a retrospective cohort study with a prospective ophthalmologic examination. Preterm and full-term participants at an age from 18 to 52 years were included. Distant corrected visual acuity (DCVA) and VRQoL were assessed in participants grouped into full-term controls , preterm participants without ROP and GA 33\u201336 weeks (group 2), GA 29\u201332 weeks (group 3), GA\u2009\u2264\u200928 weeks (group 4), non-treated ROP (group 5) and treated ROP (group 6). Main outcome measures were distant corrected visual acuity (DCVA), VRQoL and prevalence of amblyopia.p\u2009<\u20090.001) with GA, birth weight, ROP, ROP treatment, and perinatal adverse events and was poorer in both ROP groups. Visual acuity of <20/200 in the better eye was observed in two participants (4.2%) in the ROP group and one person (6.7%) in the treated ROP group. The prevalence of amblyopia increased in the ROP groups. Compared to full-term controls, visual functioning VRQoL scores were lower in preterm individuals independent of ROP while socioemotional VRQoL scores were only lower in the treated ROP group.The DCVA of the better eye correlated (Participants with postnatal ROP and its treatment showed decreased visual acuity and VRQol in adulthood, with amblyopia occurring more frequently in more preterm participants with ROP."} +{"text": "Hypoxia occurs under a variety of physiological and pathological situations with profound impact on physiological systems. The duration of hypoxia can be brief lasting several seconds to minutes or can be chronic lasting several hours to days such as that encountered at high altitude sojourn. How homeostasis is maintained during hypoxia continues to be an important question of many ongoing investigations in physiology.An adequate supply of O2 levels in arterial blood. On the other hand, chronic hypoxia such as experienced at high altitude maintains homeostasis through transcriptional activation of genes by hypoxia-inducible factors (HIFs) including HIF-1 and HIF-2. HIF-mediated activation of the Epo gene improves O2 carrying capacity by increasing erythropoietin protein. Chronic hypoxia increases formation of new blood vessels (angiogenesis) through HIF-dependent activation of vascular endothelial factor (VEGF).Acute hypoxia increases sympathetic tone, blood pressure and breathing within seconds after its onset. These rapid systemic responses to hypoxia are reflex in nature triggered by the carotid bodies, which are major sensory receptors for detecting OUnlike individuals living at high altitude, most people living at sea level encounter chronic intermittent hypoxia (CIH) because of sleep-disorder breathing manifested as obstructive sleep apnoea (OSA). Unlike continuous hypoxia, which increases both HIF-1 and HIF-2, CIH leads to imbalanced expression of HIF-1 and HIF-2, leading to increased oxidative stress and cardio-respiratory pathophysiology.2 on vascular smooth muscle and/or indirectly increasing vasodilator metabolites. In contrast, hypoxia constricts microvasculature by acting on endothelium. Hypoxic pulmonary vasoconstriction (HPV) and the resulting pulmonary hypertension represent a well-documented effects of hypoxia on microvasculature. Increased vascular permeability arising from direct effect of hypoxia on microvasculature has dire physiological consequences.Hypoxia can also act directly on the vasculature leading to disturbed vascular homeostasis. However, the effect of hypoxia depends on the blood vessel type. Hypoxia dilates large blood vessels either due to direct effects of low ORaghavan et al. address the relationship between purinergic signaling in altered vascular permeability by hypoxia. They reported that endothelial cell P2Y1 receptors mediate hypoxia-evoked endothelial barrier dysfunction and hyperpermeability, and these effects are prevented by P2Y1R antagonist. They further showed in vitro hypoxia/reoxygenation upregulates endothelial cell P2Y1receptors, which contributes to degradation of endothelial junctional proteins resulting in increased endothelial permeability. MRS 2500, a P2Y1R antagonist, inhibit P2Y1 receptors and improved endothelial barrier permeability. Malkmus et al. assessed the roles of Ca2+ regulated transient receptor potential (TRPC) proteins and pulmonary vascular remodeling in chronic hypoxia-induced pulmonary hypertension (CHPH). They hypothesized altered [Ca2+]I as one of the mediators of pulmonary vascular remodeling and assessed whether genetic deletion of TRPC 1,3,6 channels protect against CHPH in a murine model. Their findings indicate deletion of TRPC1, 3 and 6 partially protect against CHPH without affecting pulmonary vascular remodeling. Parvatikar et al. addressed efficacies of bioactive compounds from a medicinal plant (Mucuna pruriens) on cerebral ischemia and pathophysiology of brain tissue. Their work demonstrated Mucuna prureins plant extract exhibit neuroprotective actions involving downregulation of the NMDAR and tau proteins. They identified \u03b2-sitosterol as an active compound of Mucuna prureins. Lade et al. assessed the mechanisms of hypoxia induced interactions between Na+/H+ exchanger isoform 1 (NHE1) and actin filament (via p-ezrin) in pulmonary artery smooth muscle cell (PASMC). Their findings showed hypoxia increases p-ezrin and NHE1 proteins facilitating changes in PASMC phenotype and promoting vascular remodeling and develop pulmonary hypertension. M\u00fcller et al. presented OSA-related model of intermittent hypoxia (IH) in endothelial cells and its relation to vascular pathology. Their study based on patient data with OSA provides insights into inflammatory endothelial cell activation by IH which may facilitate understanding of therapeutic aspects of IH mediated vascular pathology. Moreno-Dom\u00ednguez et al. reviewed the nature of oxygen sensing in acute vasomotor response to hypoxia. They discussed two classic vasomotor responses to hypoxia including hypoxic pulmonary vasoconstriction (HPV) and hypoxic vasodilation (HVD). The review provides important translational perspectives of cardiorespiratory pathophysiology and pharmacology.The Research Topic of Frontiers in Physiology focuses on articles addressing mechanisms underlying vascular pathologies of hypoxia. The article by"} +{"text": "Purpose: To correlate postoperative optical coherence tomography (OCT) thickness changes of each retinal layer with different patterns of visual recovery after idiopathic epiretinal membrane (ERM) surgery in a cohort of patients showing no known risk factors for poor visual recovery at preoperative imaging. Methods: Best corrected visual acuity (BCVA) and OCT images were acquired preoperatively and 1 month, 3 months and 6 months postoperatively. Patients were divided into four groups according to postoperative BCVA improvement: improvement < 15 ETDRS letters (GROUP 1), immediate improvement of BCVA without further improvements at later follow-ups (GROUP 2), immediate improvement of BCVA with further improvements at later follow-ups (GROUP 3) and delayed improvement of BCVA (GROUP 4). Results: Eighty-five patients were included. GROUP1 was the only one characterized by retinal nerve fiber layer (RNFL) thickness increase and ganglion cell layer/central macular thickness (GCL/CMT) ratio decrease at 1 month and outer nuclear layer (ONL) thickness decrease at 3 and 6 months. GROUP 2 was the only one showing a decrease in GCL/inner plexiform layer (GCL/IPL) ratio at 1 month. GROUP 3 and 4 showed higher preoperative RNFL thickness compared to the other, and GROUP 4 manifested a late increase in RNFL thickness at 6 months. Conclusions: Different patterns of VA recovery are associated with specific layer thickness changes. If further confirmed, this would help detect those cases characterized by poor or delayed visual recovery despite the absence of other known imaging risk factors. Epiretinal membrane (ERM) is a disorder of the vitreoretinal interface with proliferation and metaplasia of cellular tissue on the surface of the retina along the inner limiting membrane (ILM) that is caused by a pathological separation of the vitreous cortex from the ILM ,2. ERM cWe performed a monocentric prospective analysis of patients referred to Eye Center, Humanitas Clinical and Research Center for surgical treatment of ERM between January 2017 and October 2020. All the authors reviewed the manuscript and vouched for the accuracy and completeness of the data and for the adherence of the study to the protocol (ID 1882). Informed consent and Ethics Committee approval were obtained in conformity with the Declaration of Helsinki.No variation or improvement < 15 ETDRS letters (3 Snellen lines) ,15 from Immediate (1 month after surgery) improvement of visual acuity without further improvements at later follow-ups (GROUP 2); orImmediate (1 month after surgery) improvement of visual acuity with further improvements at later follow-ups (GROUP 3); orDelayed improvement of visual acuity (GROUP 4).All considered patients underwent 25-gauge pars plana vitrectomy (PPV) with ERM and ILM peeling and air tamponade. A composition of 0.125 mg Brilliant Blue G and 0.65 mg Bromphenol Blue was used for ILM and ERM staining to assist the peeling. Surgery was performed by a single expert surgeon (J.L.V.). Postoperative treatment included topical dexamethasone + levofloxacin drops 4 times per day for 1 month and topical indomethacin drops 2 times per day for 1 month. Patients underwent a baseline examination, including best corrected visual acuity (BCVA) assessment, OCT images acquisition and slit lamp examination, and were re-examined with the same procedures at 1 month, 3 months and 6 months after surgery. Inclusion criteria were pseudophakic status and preoperative diagnosis of stage II ERM [p value < 0.05 was considered as statistically significant.Statistical analysis was conducted using SPSS software (IBM SPSS Statistics 26.0). Normality of the distribution for quantitative variables was evaluated using Shapiro\u2013Wilk test. Normally distributed variables were described using mean and standard deviation. Qualitative variables were described as number of cases over total and percentage. One-way ANCOVA for repeated measures was used to assess variations in continuous morphological parameters in the total population. Interactions between time and VA recovery pattern in change in thickness parameters were assessed with a two-way ANCOVA for repeated measures. Multinomial logistic regression was used to detect predictors in VA recovery pattern. For all omnibus tests, post hoc analysis was performed to assess individual differences. A A total of 354 eyes were screened for inclusion. Among them, 189 were excluded due to the presence of preoperative negative prognostic biomarkers: specifically, 91 showed stage III or IV ERM; 38 showed preoperative IS/OS or interdigitation zone damage or photoreceptor elongation; 25 showed preoperative cotton wool signs or localized foveal detachment; 12 showed postoperative anatomical damage such as IS/OS interruption, interdigitation zone damage or severe dissociated optic nerve fiber layer (DONFL); and the rest (65) showed overlapping retinal disease. The selection resulted in a cohort of 85 eyes of 85 patients with a mean age of 70.73 \u00b1 7.77 years and an almost equal sex distribution . Chronic kidney disease, chronic obstructive pulmonary disease and systemic arterial hypertension were present in 22.4%, 23.5% and 55.3% of the patients, respectively see .p < 0.001). Preoperative mean CMT in the total population was 483.5 \u00b1 89.7 \u03bcm. Overall, a statistically significant reduction in both central macular thickness (CMT), inner layer thickness, retinal nerve fiber layer (RNFL), ganglion cell layer (GCL) and inner plexiform layer (IPL) thickness was observed 1 month after the treatment, also accounting for covariates . Nevertheless, no further significant decrease in RNFL thickness was noted after the first postoperative month. Similarly, CMT and inner layer thickness did not vary significantly after 1 month from surgery. Differently, both GCL and IPL continued to decrease at 3 months follow-up .RNFL thickness was the most influenced among the above-mentioned parameters, changing from 92.57 \u00b1 91.07 \u03bcm before surgery to 28.7 \u00b1 25.73 \u03bcm at 1 month follow-up . Differential thickness of RNFL, inner layers and CMT from preoperative to 3 months postoperative evaluation revealed a significantly higher difference in patients from GROUP 3 and 4 compared to the others. By contrast, analyzing differential thickness from baseline to 6-month postoperative evaluation, the CMT decrease was statistically significantly higher in GROUP 3 and lower in GROUP 1 compared to all other groups. In addition, the inner layer thickness decrease was significantly higher in GROUP 3 compared to all others.The majority of patients treated belonged to GROUP 2 (35 patients), thus revealing immediate improvement in visual acuity without further improvements at later follow-ups as the most frequent VA recovery trend. By contrast, 15 patients experienced continuous progressive improvement (GROUP 3) and 15 patients showed a delayed recovery (GROUP 4). Lastly, 20 patients experienced insufficient VA improvement or worsening of initial visual function after treatment (GROUP 1). p < 0.001, see p = 0.042). In all other groups, no significant change in ONL thickness was detected during follow-up.Directly comparing the variation in retinal layer thickness during postoperative follow-ups in all the analyzed groups, different trends of change in inner layer, RNFL and ONL thicknesses were interestingly noted among groups see . With reERM is the most common type of fibrocellular proliferation found at the vitreoretinal interface ,15. SincThe first OCT thickness trend is characteristic of patients with a bad outcome (VA improvement at the end of the follow-up < 15 ETDRS letters). They manifested a significant increase in RNFL thickness at 1 month, differently from all others, and a decrease in ONL thickness at 3 and 6 months follow-up. Moreover, this group of patients was the only one characterized by a decrease in GCL/CMT ratio at 1 month follow-up D. We posThe second trend is typical of patients manifesting immediate (1 month after surgery) improvement of VA without further improvements at later follow-ups. This course was the most prevalent in our series, which is also consistent with literature on the matter . This grThe remaining two VA trends were both characterized by a delayed (after 1 month from surgery) increase in VA. One of them (GROUP 3) was the one with the best prognosis of all: these patients experienced a clinically significant VA improvement within the first postoperative month and continued to get better over the remaining follow-up. However, GROUP 4 showed an unsatisfactory outcome until the third postoperative month, manifesting > 2 lines of VA improvement at the end of the follow-up. Both these groups showed a significantly higher preoperative RNFL thickness compared to the other two groups. High preoperative RNFL thickness may thus be regarded at as an indicator of the possibility of late improvement in VA after surgery. We postulate that this late improvement could be attributable to the gradual relaxation of RNFL layer after the removal of longstanding traction. Unfortunately, not all patients included in the analysis were first diagnosed with ERM in our center, so we do not have reliable data on the duration of the disease before surgery. Another curious characteristic is that patients from GROUP 4 (delayed improvement) manifested a late (6 months) increase in RNFL thickness following the continuous degrading trend of the previous months. Limits of our study include the small sample size, the monocentric nature of the study, the strict inclusion and exclusion criteria and the lack of consideration for other ERM visual symptoms such as metamorphopsias. In conclusion, we propose a new OCT biomarker that could predict worse or delayed visual recovery in cases of absence of other known prognostic imaging risk factors for bad prognosis. If further confirmed, it would enhance the accuracy of preoperative and 1 month postoperative screening of patients at risk for reduced postoperative functional improvement."} +{"text": "Lymphedema following breast cancer surgery is a chronic and disabling complication that may lead to recurrent cellulitis. Innovative advances in treatment of breast cancer\u2013related lymphedema have evolved beyond conservative management to include vascularized lymph node transplant (VLNT). Herein, we analyzed the impact of VLNT in reducing the rate of upper extremity cellulitis in breast cancer survivors.The charts of all patients who had breast cancer and mastectomy, whose course was complicated by upper extremity lymphedema, and who ultimately underwent VLNT at our comprehensive cancer center from 2017 to 2021 were reviewed. Patients who had at least one episode of cellulitis within the year prior to VLNT and were followed for 24 months were included. Thereafter, we reviewed patients\u2019 demographics, cancer management, breast reconstructive procedures, VLNT surgeries, and infectious complications.2. Most patients had invasive ductal carcinoma (82%) and received chemotherapy (94%), including taxane-based regimens (85%), which increases the risk for lymphedema. Also, most patients received radiation therapy (86%), including to the axillary nodal basin (92%). Furthermore, most underwent unilateral mastectomy (74%) with axillary lymph node dissection (95%) followed by autologous or implant-based reconstruction (both 42%) (Table 2). VLNT was performed at a median of 92 months after mastectomy and were mainly harvested from the inguinal and gastroepiploic regions (both 32%). After VLNT, 58 (88%) patients remained infection-free throughout the 24-month follow-up period. The only factor associated with recurrent cellulitis was the need for a second lymph node transfer or lymphovenous bypass surgery, likely owing to a significant lack of lymphedema reduction (p=0.006).We included 66 patients in our study. Their median age was 57 years (Table 1). The majority were White (88%), with a mean (\u00b1 SD) body mass index of 29.4 \u00b1 6.7 kg/mThe novel approach of VLNT in the management of breast cancer\u2013related lymphedema is associated with significantly decreased recurrent cellulitis rates and should be considered as part of the infectious diseases treatment armamentarium.All Authors: No reported disclosures"} +{"text": "ARMC5 mutations occur in 20 to 55% PBMAH patients usually with more severe phenotypes. Different ARMC5 mutations might be associated with diverse phenotypes of PBMAH.Primary bilateral macronodular adrenocortical hyperplasia (PBMAH) is a highly heterogeneous disease with divergent manifestations ranging from asymptomatic subclinical Cushing syndrome (CS) to overt Cushing syndrome with severe complications. ARMC5 germline mutation , five ARMC5 somatic mutations (four novel mutations) in his right and left adrenal nodules.A 39-year-old man was admitted to our hospital with progressive weight gain and severe hypertension. He presented typical CS and its classical metabolic and bone complications like hypertension and osteoporosis. The laboratory results showed high levels of cortisol and low levels of ACTH. Low- and high-dosed dexamethasone suppression tests were negative. Contrast-enhanced computed tomography (CT) revealed multiple bilateral irregular macronodular adrenal masses. Adrenal venous sampling (AVS) confirmed that the right adrenal gland with larger nodules secreted more hormone that the left side did. Right adrenalectomy and subsequent contralateral subtotal resection were conducted. His blood pressure and CS symptoms as well as comorbidities including backache and muscle weakness improved. Whole exome sequencing identified one ARMC5 germline mutation and five different somatic ARMC5 mutations (four novel mutations) in the different nodules of the bilateral adrenal masses. AVS combined with CT imagine could be helpful to determine the dominant side for adrenalectomy. Genetic testing is important for the diagnosis and management of the patient with PBMAH.This PBMAH patient was identified with one The online version contains supplementary material available at 10.1186/s12902-023-01324-3. Primary bilateral macronodular adrenocortical hyperplasia (PBMAH) is a highly heterogeneous disease and variable levels of cortisol produced by bilateral benign adrenocortical macronodules larger than 10\u2009mm . Most paMEN1, APC and FH could result in PBMAH accompanied by other syndromic presentations such as primary hyperparathyroidism, neuroendocrine tumors, pituitary adenomas , colon polyps , leiomyomas, leiomyosarcomas and renal cancer [ARMC5 pathogenic mutations occur in 20 to 55% of patients with PBMAH [ARMC5 mutations usually have earlier onset, bigger nodules, higher cortisol levels, and more severe symptoms [ARMC5 mutation-specific [ARMC5 mutations are more often treated surgically [ARMC5 mutations might be important for the treatment decision-making. Here we report a PBMAH patient carrying one germline mutation and five somatic mutations in ARMC5.The bilateral nature of the adrenal lesions and the description of familial cases of patients with PBMAH provide support for the hypothesis of a germline genetic predisposition . Previou cancer) . ARMC5 pth PBMAH , 5. Notesymptoms . It has specific . PBMAH p2) to 71.8\u2009kg (BMI: 29.4\u2009kg/m2) and severe hypertension refractory to antihypertensive drugs for 2\u2009years (150\u2013180/100\u2013120\u2009mmHg). Physical examination showed multiple clinical manifestations of CS such as moon face, centripetal obesity, abdominal purple striae, limb edema, and muscle weakness. The hormonal work-up on admission confirmed significantly increased serum cortisol and urinary free cortisol (UFC) as shown in Table\u00a0A 39-year-old male was admitted to our hospital with progressive weight gain for 8\u2009years from 60\u2009kg BMI: 24.7\u2009kg/m4.7\u2009kg/m2ARMC5 was identified with 23 and 27 somatic mutations in his right and left adrenal, respectively was conducted on peripheral blood leukocytes and bilateral adrenal tumor tissues. A heterozygous germline mutation in ARMC5. Of note, four of these somatic ARMC5 mutations have not been identified previously. AVS confirmed that the larger mass in his right adrenal produced more cortisol. One side of total followed by contralateral subtotal adrenalectomy appeared to work well for this particular patient.PBMAH is known as an autosomal positive disease characterized with multiple nodules in bilateral adrenal like \u201ca bunch of grapes appearance\u201d . We repoThe diagnosis of PBMAH in our case mainly depend on following four criteria : (1) typARMC5 were diagnosed earlier with PBMAH with higher prevalence of overt CS and hypertension [ARMC5 mutations presented with severe CS and classical metabolic and bone complications such as hypertension and osteoporosis also support the notion that PBMAH patients with ARMC5 mutations could have severe phenotypes. Previous studies have shown that the sizes of the adrenal masses are associated with the levels of cortisol and severity of Cushing syndrome [ARMC5 mutations are correlated with cortisol hypersecretion [PBMAH is a highly heterogeneous disease with various degrees of cortisol secretion and CS. Most of PBMAH patients are not identified until overt CS occurs, patients with subclinical CS are hardly noticed unless the small enlargement of the adrenal is incidentally discovered . Therefortension . This pasyndrome , 18. Thesyndrome . Furtherecretion .ARMC5 is located at 16p11.2 with six exons and more than 80 mutations have been reported but without any identified hot spot [ARMC5 mutations occur in up to 55% of operated PBMAH patients [ARMC5 (c.1855C\u2009>\u2009T) resulting in a premature stop codon (p. R619*) which has been previously reported as a frequently mutation in Japanese patients with PBMAH [ARMC5 mutation would be needed in the background of the germline mutation [ARMC5 somatic mutations in his right and left adrenal tumors, respectively. Previous study has revealed these mutations could be nodule-specific [patients . In thisth PBMAH . Based ospecific , which mFor patients with no evidence of cortisol excess at diagnosis, active surveillance is proposed with annual clinical and biochemical assessment. On the other hand, for patients with overt CS or clinical consequences of hypercortisolism , a surgical or medical treatment should be taken into consideration. Steroid synthesis inhibitors including metyrapone and ketoconazole , mifepriARMC5 mutations has severe CS symptoms and serious complications. Somatic ARMC5 mutations might be nodule-specific. AVS combined with the sizes of the nodules is helpful in identifying the dominant side of bilateral adrenal lesion of cortisol secretion.In conclusion, PBMAH patient with Additional file 1: Supplementary Table\u00a01. Identified other 20 somatic single nucleotide variants (SNVs)/insertion-deletion (indel) mutations in the right adrenal mass.Additional file 2: Supplementary Table\u00a02. Identified other 25 somatic single nucleotide variants (SNVs)/insertion-deletion (indel) mutations in the left adrenal mass.Additional file 3. CARE Checklist of information to include when writing a case report."} +{"text": "Tibial shaft fractures are the most common long bone fractures requiring treatment. High-energy trauma often causes tibia bone injuries, causing severe complications and long-term disability due to inadequate soft tissue coverage. Tibial shaft fractures can be treated using casts, external fixators, plating, or intramedullary nails. Intramural nailing leads to faster union and reduced complications like malunion and shortening. However, patients often report subjective and objective difficulties after Surgical Instrument generation network (SIGN) nail fixation, affecting knee range of motion, quality of life, and sport activities. Tibial nails and plates are associated with increased knee pain, which negatively affects functional outcomes. No study has been conducted in a poor resource setting like Ethiopia. This study aims to assess functional outcomes of the knee and associated factors after intramedullary nailing of Tibial Diaphysial Fractures at AaBET hospital in Ethiopia.A retrospective health facility based cross-sectional study was conducted on functional outcomes of the knee and associated factors after intramedullary nailing of tibial diaphysial fractures done at AaBET hospital. A medical record review form and a structured questionnaire from patient chart and SIGN nail database collected data. The study was conducted on 151 patients registered on the SIGN nail database using a simple random sampling. Knee injury and Osteoarthritis Outcome Score (KOOS) was used to assess the knee functional outcome. Descriptive statistics such as frequency and percentage were used to summarize the results and binary logistic regression was used to describe the association between variables. P value\u2009<\u20090.05 was considered statistically significant association.The study constituted 151 patients with tibial shaft fractures; 113(74.8%) males and 38(25.2%) females with a mean age of 31.4 years, with a standard deviation of [10.5]. The prevalence of patients with good knee functional outcomes was 87(57.6%), while 64(42.4%) patients had poor knee functional outcomes. Associated factors identified include sex, age, soft tissue status, postoperative infection postoperative physiotherapy and comminuted fracture pattern.: This study determined the magnitude of knee functional outcomes revealed that more than half (57.6% ) of patients had good knee functional outcomes with identified factors increseaes odds of poor knee functional outcomes were sex, age, soft tissue injuries, post operative infection, postoperative physiotherapy and comminuted fracture patterns respectively. Therefore, Policymakers and health planners should closely monitor postoperative physiotherapy treatment courses among tibial shaft fractures treated with intramedullary nailing to increases good knee functional outcomes. The tibia is the most commonly fractured long bone in the human body, Tibial shaft fractures are defined as fractures that occur 5\u00a0cm distal to the tibial plateau and 5\u00a0cm proximal to the tibial plafond, tibial shaft is narrowest at the junction of its middle and distal third, which is the most frequent site of fracture, because of its subcutaneous nature it has poor blood supply . TraumatIn a Prospective study done at Addis Ababa University, Tikur Anbessa Hospital the incidence of Tibial fractures was higher in the distal third 942.8%) followed by middle and proximal third each 6(28.5%) . See details in Table\u00a0This study determined the magnitude of knee functional outcomes revealed that 87 (57.6%) of patients had good knee functional outcomes, while 64 (42.4%) patients had poor knee functional outcomes in Fig.\u00a0The commonest mechanism of injury in this study is road traffic accident which accounts for 96(63%) cases while fall down accident accounts for 6 (4%). More than half of the patients were injured on their right side of the extremity. A majority of the sampled patients did not have associated medical comorbidity. Among those with medical comorbidity Diabetes mellitus accounted for 10(6.6%) of cases and Hypertension accounted for 10 (6.6%) of the cases. The postoperative physiotherapy treatment course was completed by 71(47%) patients while it was only partially completed in 80(53%) patients. The mean time from injury to definitive intramedullary nailing was 5.8 days with standard deviation of [5.9]. The median time was three days with a range of 1 to 28 days see details in Table\u00a0This study had 62(41.1%) open fractures and 89(58.9%) closed fractures. Among the open fractures 26(41.9%) were Gustillo-Anderson type I, 2438.7%) type II and 12(19.3%) were Gustillo-Anderson type III. Isolated tibial fractures occurred in 41(27.2%) patients while 110(72.8%) patients had associated Fibular fracture see details in Table\u00a08.7% typeAs below Fig.\u00a0Using the independent variables which were significant on bivariate analysis; sex, categorical age, soft tissue status, comminuted fracture pattern, presence of comorbidity, postoperative infection and postoperative physiotherapy, multivariate logistic regression was carried out by including all variables to adjust for cofounders.After stepwise multiple logistic regression, it was found that females were 3.2 times more likely to have poor knee functional outcome scores than males [AOR 95% CI: 3.02 of patients had good knee functional outcomes with identified factors increseaes odds of poor knee functional outcomes were sex, age, soft tissue injuries, post operative infection, postoperative physiotherapy and comminuted fracture patterns respectively. This study reported that the commonest mechanism of injury in this study was a road traffic accident, which accounts for 63% of cases in the study area. As vehicular trauma (RTA) are most commonly associated with tibial diaphyseal fractures, nationwide awareness creation and preventive measures should be taken.Policymakers and health planners should closely monitor postoperative physiotherapy treatment courses among tibial shaft fractures treated with intramedullary nailing to increases good knee functional outcomes.High suspicion and adequate treatment plans should be provided for patients with postoperative infections.Health care providers should closely follow-up and provide appropriate management for patients with medical comorbidities.Researchers should conduct national studies to identify and address factors affecting knee functional outcome after tibial nailing."} +{"text": "The retraction has been agreed following an investigation into concerns raised by a third party, which revealed inappropriate duplications between this and another article [1]. Thus, the editors consider the conclusions of this manuscript substantially compromised.The above article, published online on 2 March 2019 in Wiley Online Library The F\u2010box protein FBXO11 restrains hepatocellular carcinoma stemness via promotion of ubiquitin\u2010mediated degradation of Snail. The retraction has been agreed following an investigation into concerns raised by a third party, which revealed inappropriate duplications between this and another article [1]. Thus, the editors consider the conclusions of this manuscript substantially compromised.The above article, published online on 2 March 2019 in Wiley Online Library The F\u2010box protein FBXO11 restrains hepatocellular carcinoma stemness via promotion of ubiquitin\u2010mediated degradation of Snail."} +{"text": "Since 2006, combined graft-versus-host disease (GVHD) prophylaxis with ATG Grafalon has been our department\u2019s base of peri-transplant supportive care. This recent retrospective study included 398 patients who underwent their first allogeneic hematopoietic stem cell transplantation after receiving a defined dose of ATG Grafalon. Our observations recorded reduced incidence of severe acute and chronic GVHD without negative impact on overall survival in a nonselected group with standard and uniform GVHD prophylaxis. Anti-T-lymphocyte immunoglobulin is a mix of polyclonal immunoglobulins isolated from rabbit serum immunized with human T-lymphoblasts from the human Jurkat cell line. It is used to reduce the risk of graft rejection and graft-versus-host disease (GVHD) in the setting of an allogeneic hematopoietic stem cell transplantation . Our pubOur retrospective study included all consecutive patients who underwent their first alloHSCT at our department between 2006 and 2020, receiving a total ATG dose of 30 or 60 mg/kg on days\u2009\u2212\u20093 to\u2009\u2212\u20091 or\u2009\u2212\u20094 to\u2009\u2212\u20092 (with FLAMSA conditioning) together with calcineurin inhibitor and methotrexate or mycophenolate mofetil. Eighty-four patients not receiving ATG for heterogeneous reasons were excluded. We analyzed (1) the cumulative incidence (CI) and severity of acute (100-day CI) and chronic GVHD (24-month CI) according to Przepiorka grading system, (2) differences depending on human leukocyte antigen (HLA) matching and conditioning regimen intensity, and (3) overall survival (OS). Risk factors influencing the development of acute or chronic GVHD were analyzed by multivariate logistic regression analysis.p<0.05) or MUD with one HLA mismatch (53%). Acute GVHD incidence was increased among patients who received a reduced intensity regimen compared to the subgroup with myeloablative conditioning . These results were confirmed also by the multivariate analysis of acute GVHD risk factors. The odds ratio that a patient who had MUD HSCT will develop acute GVHD was 1.67 compared to MSD. If administering RIC regimen, odds ratio of acute GVHD compared to a MAC regimen was 1.72.Our analysis involved 398 patients: 203 with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS), 69 with acute lymphoblastic leukemia (ALL), 44 with lymphomas, 40 with myeloproliferative neoplasms including chronic myeloid leukemia, 30 with chronic lymphocytic leukemia (CLL), and 12 other diagnoses . Myeloablative conditioning was administered to 27.6% (mainly TBI10/Cy and CyBu) and reduced intensity (RIC) in 72.4% of cases (mainly FLAMSA). One hundred seventeen patients (29%) received a graft from a matched sibling donor (MSD), 281 (71%) from an unrelated donor (MUD), 169 of them from a fully matched donor (10 out of 10), 76 with one HLA mismatch (9 out of 10). Patient characteristics are summarized in Table p<0.05) or MUD with one HLA mismatch (44.6%). Chronic GVHD incidence was increased among patients who received a reduced intensity regimen compared to patients with myeloablative conditioning .Chronic GVHD cumulative incidence at month 24 was 37.6%; extensive chronic GVHD incidence was 3.7%. As with acute GVHD cases, chronic GVHD occurred less often in related donor transplantation when compared with a full match unrelated donor transplantation . However, the 5-year OS in case of acute GVHD grade III\u2013IV was lower than in the group with acute GVHD grade I\u2013II . Ninety-five patients (24%) died due to relapse and 81 for other reasons, resulting in nonrelapse mortality of 20%.Five-year OS in the whole cohort was 55%. OS was improved when acute or chronic GVHD was present compared to OS without GVHD with our without ATG . This stAccording to our study, significantly lower incidence of acute and chronic GVHD was present in related donor HSCT. Remberger et al. analyzed the incidence of GVHD in related donor HSCT without ATG and the incidence of GVHD in MUD with ATG prophylaxis. In this study 124 patients were treated with ATG and had MUD HSCT; another 45 patients had MSD HSCT without ATG. They reported significantly lower incidence of acute GVHD grade III\u2013IV (8% vs. 25%) along with extensive cGVHD (5% vs. 40%) in the MUD group with ATG. Based on these results, authors discuss benefit of using ATG even in a related donor setting [Relative to results of several previous studies, reduced intensity conditioning had a controverse impact on GVHD. Afram et al. identified RIC regimens as one of the risk factors for cGVHD development, owing to the persistence of host-origin antigen-presenting cells . Within ATG usage in GVHD prophylaxis is detailed in many publications. A metanalysis by Kumar et al. reviews ATG prophylaxis in eight studies . ATG GraLong-term outcomes of ATG in GVHD prophylaxis were confirmed in several additional studies. Finke et al. and Walker et al. showed that cumulative incidence of relapse, NRM, and OS were comparable in groups with or without ATG , 8. AdvePatients with mild forms of GVHD achieved better overall survival probably because of the graft-versus-leukemia (GVL) effect as documented by Weisdorf et al. or BhattWithin the potential risk factors of GVHD development we confirmed by this patient cohort analysis that unrelated donors and reduced intensity conditioning regimen were the main factors that increased risk of acute and chronic GVHD incidence. Chronic GVHD also was increased in case of previous acute GVHD.In conclusion, systematic prophylaxis with ATG Grafalon reduced predominantly severe forms of acute and chronic GVHD both in related and unrelated alloHSCT. We can compare our result of OS with former publications, but other factors that contribute to OS are beyond the scope of our analysis and require further studies to discuss long-term outcomes of using ATG in GVHD prophylaxis."} +{"text": "Aripiprazole is the third generation Antipsychotic, and Dopamine serotonin system stabiliser.It is partial agonist at D2 and 5 HT1 A and antagonist at 5 HT2.Most commonly seen adverse effects are Akathesia, fatugue,insomnia and headache the major advanatage is less propensity for extrapyramidal side effects and lmetabolic side eefects.To report a case of Schizophrenia treated with Aripiprazole 15mg/day developiing occular gyric crisis which was tratment resistant.We administered Electroconvulsvive therapy, bidirectional brief pulse constant current 8 ECTS, under General anesthesia with medical fitness .Patient Showed complete resolution of Dystonia after second ECTs and Showed improvment in Pyschosis Parameterd. Assessment using Naranjo Protocol made.Electroconvulsive therapy therapy is viable alternative to manage Dystonia when medical treatment failsNone Declared"} +{"text": "Background: Anemia is a common complication among patients with cancer receiving chemotherapy and can cause significant costs to health plans.Objective: The objective of this study is to estimate the annual budget impact of drug treatment associated with treating cancer patients with anemia due to the effect of concomitant myelosuppressive chemotherapy with erythropoiesis stimulating agents (ESAs), either darbepoetin alfa (DA) once every 3 weeks (Q3W) or epoetin alfa (EA) once every week (QW), for a large US health plan in 2014.Methods: Using a patient database from a large US health plan in 2010 (n = 14 811 119), the potential CIA patient population was determined (1842 patients each per DA and EA). A budget impact of ESA treatment on this patient population in 2014 was calculated. The analysis assumed a minimum of 2 additional months of chemotherapy from initiation of the analysis. The 2014 Centers for Medicare and Medicaid Services (CMS) reimbursement rates used were: average sales price +12% of $3.68/mcg (DA) and $11.38/1000 IU (EA), and office-based injection cost of $25.08.Results: The estimated 2014 annual average drug costs per patient with CIA were $5520 (DA) and $5833 (EA). Annual average drug costs for administrations were estimated at $100 (DA) and $301 (EA) for 2014. Per member per year (PMPY) costs for patients with CIA were estimated at $5620 (DA) and $6134 (EA) for 2014. The annual total costs per CIA population (n=1842) were estimated at $10 352 629 (DA) and $11 298 798 (EA) for 2014.Conclusion: DA Q3W has the potential to provide cost savings over EA QW in terms of annual average drug cost per patient with CIA ($313 savings), PMPY costs for patients with CIA ($514 savings), and total cost per CIA population ($946 169 savings)."} +{"text": "BCR::ABL1 tyrosine kinase domain (KDM) and of which the majority of which occur at seven codons within this region. A case of CML is described in which presence of a rare D363G BCR::ABL1 KDM resulted in a suboptimal response to frontline imatinib. Switching to dasatinib resulted in achieving a sustained major molecular response that was maintained after a subsequent switch to bosutinib due to the side effects. Reporting of such cases is important for the future management of any CML patients with this rare mutation.Acquired resistance to tyrosine kinase inhibitors (TKIs) remains a therapeutic challenge in the treatment of chronic myeloid leukemia (CML). The most studied reason for TKI resistance is the acquisition of mutations within the BCR::ABL1 kinase domain mutations (KDM) extensively investigated. Mutations associated with TKI resistance have been described throughout the gene encoding, the BCR::ABL1 kinase domain, with approximately 20 mutations recurrently observed. In vitro TKI activity profiles of this common BCR::ABL1 KDM facilitate appropriate selection of subsequent TKI therapy [BCR::ABL1 KDM. More recently, next-generation sequencing (NGS) approaches have demonstrated improved limits of detection and are becoming increasingly adopted [Despite extensive advances in the therapy of chronic myeloid leukemia (CML) over the last twenty years due to the introduction of tyrosine kinase inhibitors (TKIs), a small but appreciable number of patients are either refractory or resistant to these inhibitors. The causes of resistance are myriad with acquired mutations of the adopted . Uncommo adopted \u20137.9/L, and platelet count of 278\u2009\u00d7\u2009109/L. Bone marrow aspirate and trephine biopsy showed trilineage hematopoiesis and myeloid hyperplasia with immunophenotyping demonstrating 1% myeloblasts. Cytogenetics detected the t translocation in 181/200 metaphases analysed. Quantitative reverse-transcription polymerase chain reaction detected e13/14a2 BCR::ABL1 transcripts at a high level, all consistent with a diagnosis of low-risk , chronic phase CML; the patient commenced imatinib 400\u2009mg twice daily which was well tolerated and achieved a BCR::ABL1 transcript level of 11.7% at three months approximating a warning as per European LeukemiaNet recommendations [BCR::ABL1 transcript level was 2.9% in achieving MMR in this setting. Reporting of such cases is necessary for the future, real-world management of BCR::ABL1-positive leukemia patients. The increasing application of NGS approaches is likely to improve BCR::ABL1 KDM detection allowing a timelier reconsideration of TKI therapy.The D363G mutation is exceedingly rare with only sporadic cases reported in imatinib-resistant chronic phase CML patients , 10. In"} +{"text": "Hand hygiene (HH) is a fundamental component of infection prevention and control in healthcare settings. This study aimed to identify barriers to HH according to occupational group to increase the rate of HH compliance among healthcare workers (HCWs).This cross-sectional survey was conducted in July 2018 at four university affiliated hospitals. The survey comprised seven parts with 49 items, including self-reported HH compliance, knowledge, attitudes, behaviours, barriers to HH, and improvement strategies.A total of 1,046 HCWs participated in the survey. The self-reported HH compliance rate was highest in the nurses group, followed by other HCWs and physicians . The scores regarding knowledge, attitudes, and behaviours about HH were highest in the nurses group. The nurses group also had higher scores in terms of internal and emotional motivation. The most important intervention was \u201chand sanitizer placed where necessary\u201d followed by \u201cregular HH education\u201d and \u201creward and publicize excellent HH employees/departments\u201d. The distribution of the nurses, physicians, and other HCWs was similar, and a downward bias can be seen for the physicians . In emergency situations, physicians and nurses found HH the most challenging, while other HCWs considered skin problems caused by HH products the most significant barrier. Among 12 improvement measures, around 20% of the respondents ranked \"diversify types of hand sanitizers\", \"install soap and paper towels in each hospital room\", and \"change perception through various HH campaigns\" as the top three priorities. The physician group deemed the timely reminder of HH compliance as the second most critical improvement measure .Hand hygiene and optimal hand hygiene compliance rate.Relationship between importance and achievement scores for hand hygiene improvement measures.The graph shows the importance and achievement scores for each measure on a scale of 1-5, with 1 being low and 5 being high. The measures include hand sanitizer placed where necessary (1), regular hand hygiene education (2), practical training according to the situation (3), frequent monitoring (4), department-wide feedback (5), personal feedback (6), hand hygiene information poster (7), audiovisual alarming/guidance (8), management\u2019s interest and encouragement (9), and reward and publicize excellent hand hygiene employees/departments (10).Improvement measures for barriers to performing hand hygiene based on first choice by respondents.The graph shows the percentage of respondents who selected each improvement measure as their first choice. The measures include offering different types of hand sanitizers (1), sending reminders about hand hygiene timing (2), educating patients and caregivers to promote a culture of hand hygiene among staff (3), using hand hygiene campaigns to change perceptions (4), including hand hygiene results in staff performance reviews (5), providing immediate feedback on hand hygiene observations (6), conducting regular monitoring of hand hygiene practices (7), ensuring soap and paper towels are available in all hospital rooms (8), implementing a real-name system to track hand hygiene performance (9), conducting peer-to-peer assessments of hand hygiene performance (10), strengthening hand hygiene theory education (11), and providing training for different hand hygiene situations (12).Differences in barriers hindering HH compliance and improvement plans were identified for each group. The findings suggest that targeted interventions tailored to the specific needs of different occupational groups may be effective in improving HH compliance in healthcare settings.All Authors: No reported disclosures"} +{"text": "Meningococcal vaccine uptake among adolescents remains suboptimal. This project aimed to identify barriers and motivators for vaccination in AI communities.Between 12/2022 and 2/2023, 3 live family education sessions were held within AI communities in WI and ND. Healthcare provider (HCP) participants also attended a workshop to review current vaccine recommendations. Surveys were administered before/after the workshop and family education sessions.Surveys were completed by 56 family members. Baseline confidence in vaccine safety, efficacy and benefit was low, which increased after the sessions . Parent understanding that the vaccine cannot cause disease improved from 51% to 82% (p =.001). Top reported barriers to vaccination were concern for immediate side effects and long-term side effects, and low perceived risk for IMD . Parents identified learning more about IMD (40%), friends/family getting vaccinated (19%), and a local outbreak (12%) as top motivators to seek vaccination. When asked what they thought would motivate families to seek vaccination, HCPs leading the sessions (n=11) identified learning more about IMD (82%) and friends/family getting vaccinated (27%) only. HCP confidence in counseling families was low and improved after the sessions . HCPs attending the workshop (n=28) reported discomfort contradicting patients/families as the top challenge in addressing vaccine hesitancy, and more HCPs felt confident having conversations to reduce vaccine hesitancy after the workshop (80%) than before (28%). HCP knowledge improved after the workshop, and more HCPs understood that MenACWY booster doses are not needed when initial vaccination occurs at age \u226516 (50%), and correctly identified \u201cprovider recommendation\u201d as the top literature-reported factor influencing parental vaccine decisions (50%) compared to baseline . HCP action plans for after the sessions prioritized routinely discussing vaccines at well visits (80%), providing written materials (50%) and staying current on new vaccines (50%).Parent and provider knowledge and confidence improved after the program and offered insights into community-level motivators and barriers to vaccination.All Authors: No reported disclosures"} +{"text": "High altitude pulmonary edema (HAPE) is a life-threatening form of non-cardiogenic edema which occurs in unacclimatized but otherwise normal individuals within two to four days after rapid ascent to altitude beyond 3000 m. The precise pathoetiology and inciting mechanisms regulating HAPE remain unclear.MYLK; rho family members ARGEF11, ARHGAP24; cell adhesion molecules such as CLDN6, CLDN23, PXN and VCAM1 besides other signaling intermediates. Further, several important regulators of systemic/pulmonary hypertension including ADRA1D, ECE1, and EDNRA were upregulated in HAPE. We also observed significant upregulation of genes involved in paracrine signaling through chemokines and lymphocyte activation pathways during HAPE represented by transcripts of TNF, JAK2, MAP2K2, MAP2K7, MAPK10, PLCB1, ARAF, SOS1, PAK3 and RELA amongst others. Perturbation of such pathways can potentially skew vascular homeostatic equilibrium towards altered vascular permeability. Additionally, differential regulation of hypoxia-sensing, hypoxia-response and OXPHOS pathway genes in individuals with HAPE were also observed.We performed global gene expression profiling in individuals with established HAPE compared to acclimatized individuals. Our data suggests concurrent modulation of multiple pathways which regulate vascular homeostasis and consequently lung fluid dynamics. These pathways included those which regulate vasoconstriction through smooth muscle contraction, cellular actin cytoskeleton rearrangements and endothelial permeability/dysfunction. Some notable genes within these pathways included Our data reveals specific components of the complex molecular circuitry underlying HAPE. We show concurrent perturbation of multiple pathways regulating vascular homeostasis and suggest multi-genic nature of regulation of HAPE. High altitude pulmonary edema (HAPE) is a life threatening clinical condition that occurs in otherwise healthy individuals who rapidly ascend to high altitude (above 3000 m). It is the major cause of death related to high altitude exposure. Though notable physiological and clinical studies in past two decades have underscored the importance of \u2018acclimatization schedules\u2019 in preventing HAPE, the individual susceptibility beyond acclimatization remains far from being deciphered. A prior history of HAPE Amongst the aforementioned factors, it is difficult to identify primary and secondary factors which regulate individual HAPE susceptibility. Furthermore, the involvement of inflammation in primary etiology of HAPE remains unclear, suggesting complexity of possible underlying mechanisms. It is likely that increased hydrostatic pressure, inflammatory response or a combination of both could be important The study population consisted of male sea level residents of Indian origin (n\u200a=\u200a17) who had developed HAPE after air induction (ascent by flying) to high altitude (Leh situated at 3250 m). Four volunteers were fresh inductees while the rest were re-inductees who travelled to high altitude after spending their leave period at sea level compared to other samples, however, for normalization, all samples were retained and distribution profiles obtained.Hybridization to Human Whole Genome oligochip 40K was carried out as per manufacturer\u2019s protocol and data submitted to GEO (Accession number: GSE52209). Experimental design, sampling, hybridization and data analysis were performed in compliance with Minimum Information About a Microarray Experiment (MIAME) guidelines was an indicator that it was enriched with genes. Functional Annotation Clustering was achieved using the Database for Annotation, Visualization and Integrated Discovery (DAVID) T method) 2\u2212\u0394\u0394Ct using the RQ software Confirmation of microarray results was performed on randomly selected differentially expressed genes by Real Time Polymerase Chain Reaction on a microfluidic card assay system (TLDA) of Applied Biosystems using the same samples used in microarray experiments. The card included TaqMan probes and primer sets for amplification of target genes and an endogenous control of ribosomal RNA 18S. For each sample, cDNA was prepared with High Capacity Reverse transcription kit (Applied Biosystems). qRT-PCR was performed in duplicate on cDNA samples (200 ng) in each port of TLDA card through 50\u00b0C for 2 min, 94.5\u00b0C for 10 min, 40 cycles of 97\u00b0C for 30 s and 59.7\u00b0C for 1 min on ABI PRISM 7900 HT Sequence Detection System . The data was analyzed using comparative cycle threshold method (C2) from HAPE and acclimatized individuals is presented in Supplementary The clinical parameters of heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP) and arterial oxygen saturation in peripheral blood cells of individuals with HAPE . AmongstThe hierarchical clustering of this data set distinctly separated the acclimatized lowlanders from individuals with HAPE . We infeCategorization of the differentially expressed genes based on their biological process, molecular functions and cellular component are shown in Supplementary Further analysis through DAVID revealed a number of biological processes in HAPE individuals which were significantly enriched . Some of the critical processes were those involved with oxidative phosphorylation (OXPHOS) and ATP synthesis, ion homeostasis, apoptosis, voltage-gated channel activity, iron-sulfur cluster binding, intra cellular protein transport,TNF-associated molecules, regulation of systemic arterial blood pressure, Ras-guanyl exchange factor activity amongst others. Transcripts associated with smooth muscle contraction, lipid catabolic process, calcium ion transport, actin binding and inflammatory response were also differentially expressed in HAPE individuals. The high stringency functional annotaion cluster is shown in Supplementary The differentially expressed genes in HAPE were studied for their overabundance in different pathways and are presented in TNF alpha, MAP2K2, PLA2G1B, PAK3, SOS1 , endothelin converting enzyme 1 (ECE 1), endothelin receptor type A (EDNRA), ADAM metallopeptidase domain 11 (ADAM 11), adrenergic receptor ADRA1D, angiopoeitin 4 (ANGPT4), claudin 6 (CLDN6), carbonic anhydrase IX (CA9), cytochrome P450 and voltage gated sodium channels were upregulated while the expression of angiopoeitin-like 1 (ANGPTL1), protein tyrosine kinase 2 (PTK2), peroxisome proliferator-activated receptor alpha (PPARA), Rho GTPase activating protein 24 (ARHGAP 24), thrombospondin 1 (THBS1), Heat shock protein 90 , histone deacetylase 9 (HDAC9), water channel aquaporin 12 (AQP12) and ubiquilin 4 (UBQLN4) were downregulated in HAPE compared to acclimatized controls. This data underscores prominent differences in responses of two groups of individuals to hypoxia at high altitude.In lines with the \u2018Diathesis-Stress models\u2019, we sought to determine if there was any apparent difference in the basic hypoxia responses between HAPE and acclimatized individuals. To this end, we searched for differentially expressed hypoxia responsive transcripts in our data set. Interestingly, we observed several such transcripts in our HAPE dataset . Hypoxia. Notwithstanding the limitation of single time point study which is less likely to suggest cause/consequence relationship between various events, our global expression profiling for HAPE and acclimatized individuals evoked several interesting mechanistic possibilites besides generating molecular evidence for well known physiological processes involved in HAPE There has been an ever increasing body of evidence in favour of \u2018stress failure\u2019 being one of the early phenomenon which initiates the cascade of events in HAPE. However, it has also been argued in literature that initial damage to blood-gas barrier because of \u2018stress failure\u2019 is self limiting owing to the propensity for quick closure of this barrier after pressure reduction PAK3, MAP2K2, SOS1, ARAF, NCKAP1L, PXN, (KEGG_pathway hsa:04810) were up regulated in HAPE and actin cytoskeleton regulation (has:04810) in healthy volunteers on ascent to high altitude during process of acclimatization Pulmonary capillary \u2018stress failure\u2019 is described as the mechanical breakdown of basement membrane and pulmonary capillary endothelium resulting from excessive intravascular pressure in HAPE though PECE 1). ECE 1 generates endothelin-1 (ET-1) from its inactive intermediate and its expression levels have been reported to increase in mouse carotid body under physiological hypoxia ECE1 in HAPE would logically imply higher production of ET-1 In the present study, we observed several upstream regulators of vasoconstriction in HAPE compared to the acclimatized controls; one of them being upregulated transcript of endothelin converting enzyme were also observed in HAPE individuals. Endothelin receptor type A plays an important role in hypertension, vascular remodeling, cardiac hypertrophy and coronary artery disease in vivo has been shown to inhibit HPV EDNRA and ECE 1, we speculate these genes to be key nodes in pathways regulating elevated pulmonary artery pressure in HAPE. These genes may act as potential genetic markers of high altitude acclimatization/maladaptation and these genomic loci should be scanned for identifying differences in HAPE and acclimatized individuals.Besides ADRA1D), which is potentially involved in regulation of systemic blood pressure, to be upregulated in HAPE. Adrenergic receptors mediate sympathetic responses such as smooth muscle contraction causing vasoconstriction in blood vessels including arteries to heart ADRA1D is one of the three subtypes of the \u03b11 adrenergic receptor family that act as primary catecholamine receptor in the lungs. Enhanced \u03b11 adrenergic trophic activity has been reported in hypoxic pulmonary hypertensive rats ADRA1D along with other upregulated gene transcripts of ECE1, CYP11B2, PCSK5 and EDNRA, as observed in our data set, suggests cumulative role of multiple genes in elevating pulmonary arterial blood pressure thereby supporting multi-factorial dependence for clinical presentation of HAPE.We observed up regulated expression of adrenergic alpha-1D receptor (HSP90AB3P) suggesting that many important signaling pathways may be compromised in these individuals. HSP90 is involved in HIF signaling pathway as well as many other important cellular events including protection of vital proteins of stress signal transduction pathway. Through altered binding of HSP90, activation state of nitric oxide synthase type 3 (NOS3) and nitric oxide (NO) formation was shown to be crucially regulated by polymerization state of beta actin in human platelets HSP90 in HAPE, as seen in present study, may inhibit tertiary complex formation between NOS3, beta actin and HSP90 thereby resulting in decreased NO formation in HAPE.Another important observation in HAPE individuals was the downregulation of heat shock protein 90 kDa (HDAC9) in HAPE. HDAC directly regulates HSP90 function for nuclear receptor activity HSP90 and HDAC9 transcripts were downregulated in HAPE, the mechanism of action of these regulators and their significance in cellular adaptation to high altitude hypoxia should be probed further.Another intriguing observation was down regulation of histone deacetylase 9 (TNF) in individuals with HAPE is also interesting. Our observation is in line with recent demonstration of increased TNF alpha along with IL-6, VEGF and NO in serum of HAPE patients The gene expression data obtained in the present study also showed modulation of chemokine signalling pathway, T-cell receptor signalling pathway, leukocyte endothelial transmigration, adhesion and GAP junction molecules in circulating blood cells . NotablySeveral genes implicated in oxidative phosphorylation (OXPHOS) (hsa:00190) and fatty acid metabolism (has:00071) were also observed to be differentially expressed in HAPE. At basic cellular level, hypoxia responses are known to culminate in dynamic metabolic adaptation mediated by HIF1\u03b1 stabilization: up regulation of glycolic pathway genes and modulation of cytochrome oxidase subunit genes EGLN3) which was upregulated 2.2 fold in HAPE compared to the acclimatized individuals (controls). We also observed near 1.5 fold change in EGLN1 transcript in HAPE although with insignificant p-value (0.1) (data not shown). EGLNs are a member of hypoxia inducible proline hydroxylases (PHD) family which function as cellular oxygen sensors and directly regulate HIF-\u03b1 subunit stability EGLN2 (PHD1) mRNA levels are unchanged or decreased by hypoxia, levels of EGLN1 (PHD2) and EGLN3 (PHD3) are increased by hypoxia EGLN3 transcript during HAPE in our study and in every likelihood its increased activity implies potential inhibition of HIF responsive element (HRE)-containing genes which are selectively regulated by HIF-2\u03b1 and thereby modulating basic adaptive response to hypoxia in these individuals. HAPE-specific differential expression of EGLNs observed in our data thus underscores marked differences in hypoxia sensing ability of susceptible individuals and perhaps differential activation of HIF system during high altitude responses. Notably, we did not observe statistically significant difference in the expression of HIF and von Hippel-Lindau tumor suppressor (VHL) transcripts between HAPE and acclimatized individuals.Another interesting observation in our study was differential expression of egl nine homolog 3 Click here for additional data file.Figure S2Graphical Representation of (a) enriched cellular components, molecular functions and biological processes related to the differentially expressed genes in microarray data set. (b) enriched Gene ontology terms.(PPTX)Click here for additional data file.Figure S3(A-R): Representation of sub-networks within the integrated network shown in . Individual pathways (sub-networks) interacting through common nodes generate the integrated network represented in (PPTX)Click here for additional data file.Table S1Details of subjects, sample collection and clinical parameters.(XLSX)Click here for additional data file.Table S2Minimum Information About A Microarray Experiment (MIAME) compliance of the experimental design, sampling, hybridization and data analysis.(DOC)Click here for additional data file.Table S3List of all up and down regulated genes in the microarray data.(XLSX)Click here for additional data file.Table S4Functional Annotation Clustering Table (extracted from DAVID Bioinformatic Resource).(XLSX)Click here for additional data file.Table S5KEGG, Biocarta and Panther Pathways extracted from Pathway Miner.(XLS)Click here for additional data file."} +{"text": "Complicated sternal wound infection after cardiac surgery has an incidence of 0.4 \u2013 6.9 % and mortality of 7 \u2013 80 %. The ideal reconstructive procedure is still a matter of debate.To report our experience with the laparoscopically harvested omental flap and transverse plate fixation for sternal reconstruction after complicated sternal wound infection.Between 2010 and 2011, 6 patients with type IV deep sternal wound infection underwent a sternal reconstruction with a laparoscopically harvested omental flap and transverse plate fixation. The median age of the cohort , was 72.5 years (range: 49-78 years). In 5 patients, a bilateral internal thoracic artery had been used. Considerable preoperative risk factors were present: Obesity with Body Mass Index (BMI) \u2265 33 (range: 33 \u2013 35: 3 patients); chronic obstructive pulmonary disease (COPD) without steroid therapy preoperatively (4 patients); Diabetes mellitus . Abdominal surgery had previously been performed in 4 patients. In 5 cases, the mediastinal wound was prepared with negative pressure wound therapy following surgical debridement. An internal fixation of the sternum was made by titanium locking plates with sternal and rib-to-rib fixation. The postoperative course of these patients was followed by clinical follow-up.Early postoperative sternal stability was seen in all 6 patients. The 30-day perioperative mortality rate was zero, with an overall survival of 100% until today. Postoperatively no superficial or deep surgical site infections (SSI) were appreciated. Follow-up ranged from 24 to 41 months (median: 28 months).Combination of a laparoscopically harvested omental flap and transverse plate fixation can contribute to a successful outcome following complicated sternal wound Infection and deserves serious consideration, regardless of the co-morbidity or previous abdominal surgery."} +{"text": "Previous genetic studies have demonstrated that natal homing shapes the stock structure of marine turtle nesting populations. However, widespread sharing of common haplotypes based on short segments of the mitochondrial control region often limits resolution of the demographic connectivity of populations. Recent studies employing longer control region sequences to resolve haplotype sharing have focused on regional assessments of genetic structure and phylogeography. Here we synthesize available control region sequences for loggerhead turtles from the Mediterranean Sea, Atlantic, and western Indian Ocean basins. These data represent six of the nine globally significant regional management units (RMUs) for the species and include novel sequence data from Brazil, Cape Verde, South Africa and Oman. Genetic tests of differentiation among 42 rookeries represented by short sequences and 40 rookeries represented by long sequences supported the distinction of the six RMUs analyzed as well as recognition of at least 18 demographically independent management units (MUs) with respect to female natal homing. A total of 59 haplotypes were resolved. These haplotypes belonged to two highly divergent global lineages, with haplogroup I represented primarily by CC-A1, CC-A4, and CC-A11 variants and haplogroup II represented by CC-A2 and derived variants. Geographic distribution patterns of haplogroup II haplotypes and the nested position of CC-A11.6 from Oman among the Atlantic haplotypes invoke recent colonization of the Indian Ocean from the Atlantic for both global lineages. The haplotypes we confirmed for western Indian Ocean RMUs allow reinterpretation of previous mixed stock analysis and further suggest that contemporary migratory connectivity between the Indian and Atlantic Oceans occurs on a broader scale than previously hypothesized. This study represents a valuable model for conducting comprehensive international cooperative data management and research in marine ecology. Caretta caretta) is a globally distributed species with a complex life cycle. After leaving their natal beaches, hatchlings swim into major ocean surface currents and may be transported across entire ocean gyres as epipelagic, oceanic juveniles Marine vertebrates with cosmopolitan distributions often exhibit high dispersal and weak population structure across large spatial scales. Notable exceptions include groups that exhibit reproductive philopatry despite extensive migrations and dispersal potential, such as salmonids Females exhibit natal philopatry, returning to nest in the region where they hatched At the global level, nine regionally significant nesting aggregations have been recognized as Regional Management Units (RMUs) based on genetic, demographic, geographic, and oceanographic considerations: 1) Northwest Atlantic Ocean, 2) Southwest Atlantic Ocean, 3) Northeast Atlantic Ocean, 4) Mediterranean Sea, 5) Southwest Indian Ocean, 6) Northwest Indian Ocean, 7) Southeast Indian Ocean, 8) North Pacific Ocean, and 9) South Pacific Ocean with a tenth putative RMU proposed for the Northeast Indian Ocean for which genetic and biological data are lacking Regional reassessments of population structure using longer control region fragments (760 to 817 bp) and representing previously unsampled rookeries have demonstrated that additional MU designations were warranted This research was approved by Institutional Animal Care and Use Committees at the University of Florida (201101985) and the University of Georgia (A201201-025-R1). Georgia samples were collected under Georgia Department of Natural Resources permit 29-WJH-13-37. Florida samples were collected under Florida Fish and Wildlife Conservation Commission permits MTP-016, MTP-130, and MTP-135. This work was conducted under SISBIO permit 14122-1 from the Brazilian Ministry of the Environment, and samples were exported under CITES permit 11BR006778/DF. Samples were collected in South Africa under authority of the Department of Marine Affairs permit RES2010-44 and RES2010-55 and exported under CITES permit 106682. Samples were imported into Spain (University of Barcelona) under CITES permits ESBB00601/03-I, TR18080303092, 106126/3423 and 1186. Samples were imported into the United States under CITES permits 13US724540/9 (Archie Carr Center for Sea Turtle Research) and 09US844694/9, 10US844694/9 (NOAA - Southwest Fisheries Science Center).Haplotype counts representing 380 bp and \u223c800 bp control region sequences were taken from the literature or generated from novel samples . New sam2, 0.2 mM dNTPs, 0.5 unit of Taq DNA Polymerase, and approximately 10 to 30 ng of genomic DNA. PCR cycling parameters were as follows: 95\u00b0C for 3 min; 35 cycles of 95\u00b0C for 30 s, 55\u00b0C for 60 s, 72\u00b0C for 30 seconds; and a final extension of 72\u00b0C for 10 min. PCR products were purified by adding 2 \u00b5l of ExoSAP-IT\u00ae (USB Corporation) to 7 \u00b5l of the PCR reaction and incubated according to manufacturer\u2019s instructions. The mtDNA amplicons were sequenced using ABI BigDye v3.1 (PE Applied Biosystems) and an ABI 3130xl or 3730xl DNA Analyzer with LCM15382 and H950g. Samples processed at the University of Georgia (Cape Verde) were sequenced using LCM15382 and an internal sequencing primer CC443, (TGATCTATTCTGGCCTCTG). Negative controls were included in each batch of PCR amplifications and sequencing reactions to detect contamination.Genomic DNA was extracted using standard phenol-chloroform isolation or the DNeasy blood and tissue kit (QIAGEN) following standard protocols. Polymerase chain reaction (PCR) amplifications of an \u223c800 bp fragment of the mitochondrial control region were carried out using primers LCM15382 and H950g http://accstr.ufl.edu/accstr-resources/cclongmtdna.pdf). Original, short haplotypes received consecutive number designations based on the 380 bp sequence. Haplotypes based on the \u223c800 bp fragment retain their original 380 bp designations and receive additional numeral suffixes to reflect any novel polymorphisms detected within the expanded sequences. Samples producing novel or ambiguous sequences were subjected to a second round of DNA extraction, PCR amplification, and sequencing for verification. Novel haplotypes were deposited with GenBank and ACCSTR. An unrooted parsimony network was created using the program TCS www.seaturtle.org.Sequences were aligned, edited, and compared to previously described haplotypes using the program Sequencher 4.2 (Gene Codes Corporation). Sequences were assigned haplotype designations after nomenclature published on the Archie Carr Center for Sea Turtle Research (ACCSTR) website (STh), pairwise FST comparisons, pairwise exact tests of population differentiation, and genetic distance-based analysis of molecular variance (AMOVA) were conducted using the software Arlequin version 3.5 ST comparisons, and exact tests of population differentiation with p-values less than 0.05 considered significant. Exact tests of population differentiation were conducted with 100,000 permutations and 10,000 dememorization steps ST/(1-FST), and geographical distances were untransformed shortest sea distances between rookeries that accounted for major continental coastlines Population structuring at the study-wide scale was initially tested by considering the relative magnitude of barriers to gene flow as implemented in BARRIER based on frequency-based FST values from comparisons involving Cuban and Tunisian haplotype profiles were generated from a separate analysis so that the differences between values generated from 380 and \u223c800 bp haplotype data for remaining rookeries could be attributed solely to differences in haplotype resolution. Following pairwise FST comparisons and exact tests of population differentiation, proximal sample sites that were not significantly different were pooled for further analyses. Rookery clustering was also validated in an AMOVA framework by testing alternative management grouping scenarios in order to maximize FCT and minimize FSC. To minimize bias in the case of incorrect pooling decisions, haplotype frequencies were weighted for each proposed management unit based on the relative size of individual rookeries comprising them (based on average nest counts or nest count ranges). Significance of the final round of pairwise FST comparisons and exact tests of population differentiation were adjusted using a false discovery rate correction with a table-wide \u03b1 of 0.05 Because only short sequences were available from Cuba and Tunisia, the reported FLepidochelys as outgroups. Indels were coded as binary and included in analyses as a separate partition Sequences were truncated to an 822 bp alignment for phylogenetic analyses including Pacific loggerhead turtle haplotypes and Lepidochelys kempii, GenBank JX454981) and two olive ridleys (Lepidochelys olivacea) were included as outgroup taxa (GenBank AM258984 and JX454991). Fossil-derived divergence time spans at two nodes were used for calibration: the Caretta-Lepidochelys split at 12\u201320 million years before present (MYBP) Lepidochelys olivacea-Lepidochelys kempii split at 4.5 to 5 MYBP Divergence times were explored using Bayesian molecular clock frameworks implemented in BEAST v1.7.4 Excluding the Pacific haplotypes, 70 variable positions resolved 59 haplotypes for the expanded control region sequences , which iBayesian phylogenetic analysis indicated the presence of two major loggerhead lineages globally . The deeST comparisons and 78 exact tests of population differentiation were significant . Haplotype frequencies under a BRZN and BRZS pooling scenario (with a break between BAH and ESP) were also significantly different with 800 bp haplotypes .In the initial pairwise tests comparing Brazilian rookeries, there was indication of structure between RIO and the rookeries to the north but not between the two central Brazilian rookeries . HaplotyST\u200a=\u200a0.027, p\u200a=\u200a0.009; exact test p\u200a=\u200a0.012), indicating a lack of strong structure among island rookeries comprising the Northeast Atlantic RMU . The geographic distribution of haplotype frequencies among RMUs accounted for the majority of genetic partitioning, although additional structuring among rookeries within some of the RMUs was evident, particularly within the Northwest Atlantic RMU (Northwest Atlantic RMU- 1) NUSA, 2) CEFL, 3) SEFL, 4) DRSL, 5) QRMX/SWC, 6) KEY, 7) CSK, 8) NWFL; Southwest Atlantic RMU- 8) BRZN, 9) ESP, 10) RIO; Mediterranean Sea RMU- 11) CAL, 12) WGRC, 13) CRT, 14) DYDL, 15) TKW, 16) EMED, and 17) LIBY/TUN.Application of up to ten barriers in the BARRIER analysis differentiated among the six RMUs as well as indicating structure within the Northwest Atlantic and Mediterranean RMUs. The Mantel test detected the presence of very weak but significant isolation by distance . There was significant differentiation among discrete rookeries based on expanded haplotype frequencies analysis of the mitochondrial genome indicated two deeply divergent global lineages among loggerhead turtles. One major haplogroup was represented by RFLP haplotype B (now sequence CC-A1) from the southeastern USA, RFLP C (sequence CC-A4) from Brazil, and RFLP F (sequence CC-A11) from Oman. The second major haplogroup was represented by RFLP haplotype D (sequence CC-A2) from the southeastern USA, South Africa, and Greece; RFLP G from Japan (sequence CCP2 and CCP3), and RFLP H (sequence CCP1 and CCP5) from Australia Lepidochelys) haplotypes suggested that the northern Indian Ocean represented the ancestral refugium from which all extant lineages radiated Dermochelys coriacea) that currently nest circumglobally The CCP haplotypes (haplogroup IA) currently found in eastern Indian and western Pacific Ocean rookeries appear to be the oldest extant lineages in the species having diverged from the remaining clade I lineages approximately 2.7 MYBP. An Indo-Pacific origin for at least one of the major loggerhead turtle clades would be consistent with phylogeographic scenarios proposed for other marine turtle species. Analysis of global ridley turtle where haplotype CM-A8 is the sole representative of its clade in South Mozambique Channel rookeries in the Southwest Indian Ocean Invasion of the Atlantic by Indo-Pacific lineages of marine organisms via southern Africa is well established Sphyrna lewini) Inter-oceanic exchange of loggerhead turtle lineages likely occurred multiple times Haplotype compositions for marine turtle rookeries are widely used for two primary purposes: 1) to inform recognition of demographically partitioned rookery MUs for the purposes of conservation and management, and 2) to provide baseline frequencies for MSA of foraging aggregations. Ideally, these would be accomplished via a singular dataset, but even with the current expanded sequences, marker resolution remains a significant impediment to fully realizing this goal. Furthermore, investigators may be conducting MSA at varying spatial scales and in different regions, which may shift priorities for rookery splitting and lumping. For example, analysis of juvenile loggerheads in the western Mediterranean may be conducted hierarchically, first assigning Atlantic versus Mediterranean origins for all individuals where possible see followedPrevious analysis of 380 bp haplotypes indicated that the Brazilian nesting aggregation could be divided into northern (Sergipe and Bahia) and southern (Esp\u00edrito Santo and Rio de Janeiro) MUs based on the presence/absence of haplotype CC-A24 Observed genetic differentiation supported the genetic distinctiveness of the Northeast Atlantic, Southwest Indian, and Northwest Indian Ocean RMUs and the presence of at least 18 MUs across the remaining RMUs considered in this study, defining 21 population units for loggerhead turtles globally exclusive of eastern Indian Ocean and western Pacific Ocean rookeries. Within the Northwest Atlantic RMU, eight rookery clusters were evident: 1) northern MU (NUSA), 2) central eastern Florida (CEFL), 3) southeastern Florida (SEFL), 4) Cay Sal, Bahamas and the Dry Tortugas, Florida (DRSL), 5) Quintana Roo, Mexico (QRMX) and SWC, 6) southwestern Florida (KEY), 7) central western Florida (CSK), and 8) northwestern Florida (NWFL). Within the Mediterranean RMU, there was support for at least seven rookery clusters: 1) Calabria, Italy (CAL), 2) Libya (LIBY), 3) western Greece (WGRC), 4) Crete (CRT), 5) Dalyan and Dalaman, Turkey (DLY/DAL), 6) western Turkey (TKW), 7) and the remaining eastern basin rookeries (TKM/TKE/ALA/AKA/LEB/ISR). No structure was detected among islands comprising the Northeast Atlantic RMU, and no genetic variation was detected among individuals within the Southwest Indian Ocean and Northwest Indian Ocean RMUs. Sampling effort should be expanded for Oman to further explore haplotype diversity given the small sample size and the vast numbers of females that nest there SC value from significant to insignificant in the AMOVA Click here for additional data file.Table S2Variable positions for expanded control region loggerhead turtle haplotypes recorded from global rookeries.(XLS)Click here for additional data file.Table S3Mitochondrial DNA control region haplotype counts based on 800 bp sequences and haplotype diversity for loggerhead turtle rookeries in the Mediterranean Sea, Atlantic, Southwest Indian Ocean, and Northwest Indian Ocean basins.(XLS)Click here for additional data file.Table S4Pairwise comparisons based on 800 base pair (bp) control region haplotype frequencies for loggerhead turtle rookeries in the Mediterranean Sea, Atlantic, Southwest Indian Ocean and Northwest Indian Ocean.(XLS)Click here for additional data file.Table S5Pairwise comparisons based on 800 base pair (bp) control region haplotype frequencies for pooled loggerhead turtle rookeries in the Mediterranean Sea, Atlantic, Southwest Indian Ocean and Northwest Indian Ocean .(XLSX)Click here for additional data file.Table S6Pairwise comparisons of RMU and proposed MU haplotype frequency differentiation based on 800 bp haplotypes.(XLSX)Click here for additional data file.Table S7Pairwise comparisons of 380 bp haplotypes from Reis et al. 2010 and the present study for Brazilian loggerhead rookeries.(XLS)Click here for additional data file.Table S8AMOVA results for alternative management scenarios to test Mediterranean groupings based on 14 core RMUs and MUs that were held constant in their isolation: 1) NUSA, 2) CEFL, 3) SEFL, 4) DRSL, 5) QRMX, 6) KEY, 7) CSK, 8) NWFL, 9) BRZN, 10) RIO, 11) CPVD, 12) CAL, 13) NAT 14) MAS.(XLS)Click here for additional data file."} +{"text": "Studies using functional inhibition of neogenin resulted in a significant attenuation of inflammatory peritonitis. In studies employing bone marrow chimeric animals we found the hematopoietic presence of \u2212/\u2212Neo1 to be responsible for the attenuated inflammatory response. Taken together our studies suggest that the guidance receptor neogenin holds crucial importance for the propagation of an acute inflammatory response and further define mechanisms shared between the nervous and the immune system.Neuronal guidance proteins (NGP) were originally described in the context of axonal growth and migration. Yet recent work has demonstrated that NGPs also serve as guidance cues for immune competent cells. A crucial target receptor for NGPs during embryonic development is the neogenin receptor, however its role during acute inflammation is unknown. We report here that neogenin is abundantly expressed outside the nervous system and that animals with endogenous repression of neogenin ( Inflammatory conditions such as sepsis are marked by acute inflammatory changes within the affected tissue sites. During this inflammatory response immune competent cells extravasate from the vascular space and migrate along a chemotactic gradient created by chemokines to reach the site of inflammation An important target receptor to mediate NGP function in the nervous system is the neogenin receptor, a type I transmembrane protein that significantly influences axonal guidance and neuronal survival. Neogenin is expressed on maturing neuronal populations, including those in the developing cortex, midbrain and hindbrain \u2212/\u2212Neo1) demonstrated decreased inflammatory changes in the peritoneal cavity following zymosan A (ZyA) induced peritonits. Functional inhibition of neogenin resulted in a significant attenuation of this inflammatory response. Studies employing chimeric animals identified an important role of hematopoietic neogenin for the promotion of an acute inflammatory response.Based on the fact that NGPs hold important function outside the CNS we pursued the role of neogenin during an acute inflammatory response. We found that animals with endogenous repression of neogenin to induce peritonitis. In a subset of experiments animals were injected with either functionally inhibiting anti-Neo1 antibody or 1 \u00b5g IgG control antibody. Recruited leukocytes were obtained 8 hours later by peritoneal lavage with calcium- and magnesium-free ice-cold PBS solution (5 ml) containing 10 U/ml unfractionated heparin. Collected cells were washed, resuspended in 2 ml of Hanks' balanced salt solution, counted and cytospin samples were prepared. All reagents used were endotoxin-free.5\u2032-GCT GCT CTC ACA GTC AAT GG -3 and 5- GCA TAA CCT CGG ACC ACA AT -3 antisense primer. Murine \u03b2 -actin expression was evaluated with: sense 5\u2032-CTC TCC CTC ACG CCA TCC TG-3\u2032 and antisense 5\u2032-TCA CGC ACG ATT TCC CTC TCA G-3\u2032.Murine transcriptional analysis of Neogenin mRNA levels was performed using sense primer For Western blot analysis animal samples were homogenized, normalized for protein levels and applied to SDS containing polyacrylamid gels. Antibodies used for Western blotting included monoclonal anti\u2013Neogenin for murine neogenin analysis. Actin was stained using monoclonal rabbit anti- Actin . Blots were washed, and species-matched peroxidase-conjugated secondary antibody (Santa Cruz Biotechnoloy) was added. Labeled bands from washed blots were detected by enhanced chemiluminescence (Amersham Pharmacia Biotech).Blood was collected, pooled and incubated with erythrocytes lysing solution for 5 min at room temperature. Cells were then incubated with PE conjugated rabbit anti-Neo1 (Santa Cruz 15337), PerCp conjugated anti-CD 45 (BD Bioscience 5572) or isotype matched control antibody. Flow-cytometry was performed using BD FACS Canto\u2122 II employing BD FACS Diva\u2122 Software.Following peritonitis, mice were killed and tissues fixed in 3,75% formaldehyde solution. Tissues were then embedded in paraffin and stained with H&E. Pictures of tissue sections were taken using Leica Microscope (DM-RBE).137Cs source. Immediately after irradiation, 107 BM cells/recipient were injected in 0.2 ml 0.9% sodium chloride into the jugular vein. Bone marrow was transferred from characterized Neo1\u2212/\u2212 mice to WT animals and vice versa. To control for non specific radiation effects bone marrow was transplanted from WT\u2192WT and Neo1\u2212/\u2212\u2192Neo1\u2212/\u2212mice. Success of transplantation was controlled by identifying switch of hematopoetic Neo1 expression through real time PCR and Western blot analysis.To define the contribution of the myeloid and tissue specific Neo1 we generated bone marrow chimeric mice as described previously Tumor necrosis factor \u03b1 (TNF- \u03b1), interleukin (IL)-1\u03b2, IL-6, and KC were measured in the peritoneal lavage of experimental animals by standard ELISA (R&D Systems).t test was used where appropriate. A value of P<0.05 was considered significant.All values are expressed as mean \u00b1 SEM. Using Kolmogorov-Smirnov test we could show that the measured values were approximately normally distributed. Statistical significance was determined using one-way ANOVA followed by Bonferroni's multiple-comparison test. Student's We initially addressed the question whether Neo1 is expressed in murine organs outside the central nervous system to validate our model subsequently used. We found robust mRNa and protein expression of Neo1 within several murine organs . To conf\u2212/\u2212Neo1 mice in a model of ZyA induced peritonitis. 8 hours following ZyA injection \u2212/\u2212Neo1 mice demonstrated significantly reduced number of cells in the peritoneal lavage compared to WT animals , IL-1\u03b2 , IL-6 and KC , compared to WT animals and appropriate IgG isotype as control. Following this we found a significant reduction of the cell number within the peritoneal fluid of anti-Neo1 injected animals compared to IgG controls demonstrated a significantly decreased cell number in peritoneal fluid compared to the animals with haematopoetic neogenin expression (haemat. +/+Neo1) when exposed to ZyA s that were initially described in the context of axonal migration but were demonstrated to also serve as endogenous guidance cues for leukocyte migration. Neogenin is an important target receptor for NGPs during axonal growth and neuronal development yet its role during acute inflammation to date was not elucidated. We report here that neogenin is an important propagator of an acute inflammatory response during the initial stages of inflammatory peritonitis. Neogenin was initially described by Vielmetter et al. when searching for proteins involved into neurite extension and neuronal signaling The functional role of neogenin signaling is well described in the context of neuronal development. During axonal growth netrin-1 binds to neogenin which results in a chemoattractive signal, but when binding to the repulsive guidance molecule A (RGM-A) neogenin serves as a chemorepulsive receptor for neuronal migration In conclusion we identified a previously uncharacterized function for neogenin outside the CNS. Our findings suggest that signaling through the neogenin receptor holds pro-inflammatory potential and intensifies inflammatory tissue changes. Furthermore, the findings in this study highlighted the importance of hematopoietic neogenin during acute peritoneal inflammation, and that a functional inhibition of neogenin dampens an acute inflammatory response. The identification of endogenous factors that influence infection and inflammation has important medical applications and may potentially be used to enhance regeneration and recovery following inflammatory changes in the future.Figure S1Chimeric animals and controls exposed to intraperitoneal NaCl injection.A) Cell count within the peritoneal fluid in chimeric animals and controls 8 hours following intraperitoneal NaCl injection B) Myeloperoxidase (MPO) activity in peritoneal lavage C) Protein content in peritoneal lavage of chimeric animals and controls 8 hours following intraperitoneal NaCl injection. .(TIF)Click here for additional data file.Figure S2Histological assessment of chimeric animals and controls exposed to intraperitoneal NaCl injection. Representative histological analysis of the peritoneum, the mesenterial fat and cytospin samples of the peritoneal lavage of chimeric animals and controls 8 hours following intraperitoneal NaCl injection. Sections prepared with hematoxylin-eosin staining .(TIF)Click here for additional data file.Figure S3Cytokine concentration in chimeric animals and controls exposed to intraperitoneal NaCl injection.A) TNF-\u03b1 concentration in peritoneal lavage B) IL-1\u03b2 concentration in peritoneal lavage C) IL-6 concentration in peritoneal lavage D) IL-8 concentration in peritoneal lavage of chimeric animals and controls injected with zymosan A 8 hours following intraperitoneal NaCl injection .(TIF)Click here for additional data file."} +{"text": "Somatic activity of Pxt is required, as RNAi knockdown of pxt in the follicle cells recapitulates both the temporal expression and eggshell defects. One of the temporally regulated genes, cyp18a1, which encodes a cytochromome P450 protein mediating ecdysone turnover, is downregulated in pxt mutant follicles, and cyp18a1 mutation itself alters eggshell gene expression. These studies further define the molecular program of Drosophila follicle maturation and support the idea that it is coordinated by lipid and steroid hormonal signals.Drosophila ovarian follicles complete development using a spatially and temporally controlled maturation process in which they resume meiosis and secrete a multi-layered, protective eggshell before undergoing arrest and/or ovulation. Microarray analysis revealed more than 150 genes that are expressed in a stage-specific manner during the last 24 hours of follicle development. These include all 30 previously known eggshell genes, as well as 19 new candidate chorion genes and 100 other genes likely to participate in maturation. Mutations in Drosophila ovarian follicles mature during their final day of development into a functional oocyte encased in a multi-layered shell capable of protecting and nurturing the developing embryo Cp36 (FBgn0000359) and Cp38 (FBgn0000360) are active only during stages 11\u201312, \u201cmiddle\u201d chorion genes such as Cp19 (FBgn0000356) and Cp16 (FBgn0000356) during stages 13\u201314, and \u201clate\u201d chorion genes such as Cp18 (FBgn0000357) and Cp15 (FBgn0000355) during only stage 14. Chorion genes are also spatially regulated within the follicle Eggshell proteins have been identified by electrophoresis (reviewed in pxt gene (FBgn0261987) has been proposed to encode a Drosophila cyclooxygenase 2 and PGF2alpha can overcome the effects of the inhibitors in mice 2alpha rescues development in zebrafish, silkmoth, and Drosophila Prostaglandins (PGs), short-acting lipid hormones derived from arachidonic acid by the action of cyclooxygenase, play multiple roles in mammalian follicle development pxt mutation and cyp18a1 misexpression alter the timing of chorion gene expression and disrupt eggshell morphogenesis. Our studies support the view that a Pxt-dependent signal acts along with ecdysone to coordinate eggshell production.Here we have used characteristic temporal regulation to identify new eggshell genes and to identify genes involved in multiple aspects of egg maturation. One of these, Cyp18a1 (FBgn0010383), was recently shown to encode a cytochrome P450 protein and to mediate the breakdown of the steroid hormone ecdysone pxt expression was obtained from the Vienna Drosophila RNAi Center (transformant identification number 14379). The RNAi line was crossed with c355 and t80 GAL4 driver lines (Bloomington Drosophila Stock Center) to assess the roles of Pxt in the soma during follicle development. RNAi crosses and progeny were maintained at 27.5\u00b0C. The Cyp18a1 allele utilized was d01488 .Fly stocks were maintained at 20\u201325\u00b0C on standard cornmeal-agar-yeast food. pxt[f01000], pxt[EY03052] and UASp Pxt strains were described previously Drosophila version 2.0 Affymetrix chips. Microarrays were performed at least twice using independently prepared RNA samples for each stage and genotype. Microarrays were normalized to the same total expression, which is believe to change little during egg maturation due to the abundant RNA content of the germ cells. Correction factors were quite small (less than 50%) and were negligible except for highly expressed genes (top 10%). For a gene to be scored as temporally regulated, its transcript level had vary more than 4-fold during maturation (stages 9\u201310A to S14), and to show peak expression above 150. However, most of the selected genes far exceeded these parameters. All data is MIAME compliant and the raw data has been deposited in GEO.Specific staged follicles were hand dissected in room temperature Grace's medium (Lonza). The media was removed, and the tissue was ground with a plastic pestle (Kontes) in 100 \u00b5l of Trizol (Invitrogen), quick frozen in liquid nitrogen, and stored at \u221280\u00b0C until enough tissue was isolated. RNA was extracted according to the manufacturer's instructions. The Johns Hopkins Microarray Core Facility performed the microarray analyses using 10 \u00b5g of RNA to probe A subset of the microarray results was verified by semi-quantitative RT-PCR. Stage-specific follicle RNA was isolated, as described above. cDNA was synthesized using Reverse Transcription Kit (Promega), and 0.25\u20133 \u00b5L of cDNA was used per PCR reaction with primers for specific genes. Control reactions were performed at identical cDNA concentrations and PCR conditions. Primer sequences are available upon request. PCR reactions were analyzed using agarose gel electrophoresis and imaged with a UVP BioDocIt system.For all embryo collections, flies were allowed to lay eggs on wet yeasted molasses plates or grape juice plates for 30 minutes, the plate was replaced with a freshly yeasted plate and flies were allowed to lay eggs overnight. To quantify chorion and dorsal appendage defects, the laid eggs were examined on the plates under a dissecting scope. To assess for vitelline membrane defects a neutral red dye uptake assay was performed essentially as described Whole mount samples were fixed with 4% paraformaldehyde for 10 minutes and processed using standard procedures Dissected stages of follicle development and laid eggs were imaged in Grace's media (Lonza) on an Olypmus SZ61 dissecting scope with a DP25 color camera and DP2-BSW software.z-stacks produced using ImageJ and cropped/rotated in Photoshop.Immunofluorescent samples were imaged with a 20\u00d7 (NA 0.7) lens on a CARV- spinning disc confocal with a Hamamatsu EM-CCD Digital ImagEM Camera (C9100-13). All confocal images are projected Since known eggshell genes are each expressed during characteristic temporal intervals of follicle development, we searched for additional temporally regulated follicle genes. RNA was prepared from groups of 100\u2013300 hand-isolated ovarian follicles at 4 stages that span the period of eggshell production: S9-10A, S10B, S12, and S14 . Each RNThe results observed with the 30 genes encoding known eggshell proteins suggested that this approach was both accurate and highly sensitive . TranscrKnown eggshell protein genes display several distinctive characteristics in addition to stage-specific expression. In many cases they are present in gene clusters, are highly expressed but only in the ovary, and encode proteins that undergo relatively rapid evolutionary change. Based on these characteristics, we identified 19 of the new stage-specific genes as likely to encode eggshell proteins .CG13998 (FBgn0040949), which is expressed in a pattern consistent with a role in vitelline membrane formation. At the 4B4 gene cluster, near the eggshell genes femcoat (FBgn0041252) and CG15570 (FBgn0029697) CG32774 , CG15571 (FBgn0029696), and HDC17346 (FBsf0000016774), a highly expressed RNA with an uncertain gene model. Thus, at least 5 chorion proteins appear to be encoded at this locus. Additionally, in region 22A2, adjacent to the known chorion gene CG31928 (FBgn0051928) CG31926 (FBgn0051926) and CG31661 (FBgn0051661). All three of these proteins are predicted to encode aspartic proteases. Within the 30B minor amplified domain CG13114 (FBgn0032127) CG13113 (FBgn0032126) undergoes stage-specific expression, but at a quantitatively lower level. Finally, the genes CG15721 (FBgn0030438), CG32642 (FBgn0052642), CG32644 (FBgn0052644) and CG12716 (FBgn0030439) constitute an entirely new cluster of putative eggshell genes in region 11D5. As with the other chorion clusters, the genes are expressed during similar but not identical intervals. CG32644 encodes another mucin-related protein (Mur11D), while CG15721 and CG12716 contain protease inhibitor domains, including a follistatin-like domain.Eleven of these genes reside within clusters, like most previously characterized chorion genes . For exaCG32602 in situ to 12E as strongly as to the major chorion cluster at 7F CG1077 (FBgn0037405) is predicted to possess anti-microbial activity. Another, CG32972 (FBgn0028905) has a fasciclin-like domain and a region with homology to yolk granule proteins. All the candidate chorion proteins were expressed specifically in the ovary , are elevated 2\u20133 fold, an effect confirmed by RT-PCR (vm34C (FBgn0003983), vm26Aa (FBgn0003979), and impL2 (FBgn0001257), shut off RNA levels more slowly than in wild type (yellow-g (FBgn0041709) is detected prematurely in stage10B f01000pxt follicles, and persists into stage 14. CG13114, CG4009 (FBgn0038469), and cp18 appear prematurely in stage10B, while the onset of CG13084 (FBgn0032788), cp16, femcoat, CG15570 and CG15571 expression is also shifted earlier and/or prolonged. The expression of other genes, including many non-chorion genes was also altered in timing and amount in pxt mutant follicles and rala . Thus, loss of Pxt alters the temporal program of gene expression associated with egg maturation.Loss of Pxt substantially changed the temporal program of gene expression during late oogenesis. Transcripts from one of the early chorion genes, y RT-PCR . Severalild type . Transcrollicles . Pxt mutpxt mutant eggs to determine if the expression changes observed above were associated with eggshell defects. To assess the vitelline membrane integrity, we examined whether dechorionated eggs are impermeable to neutral red dye. 41% (n\u200a=\u200a179) of laid f01000pxt eggs and 34% (n\u200a=\u200a240) of laid EY03052pxt eggs took up dye, indicating a vitelline membrane defect, compared to only 1.7% (n\u200a=\u200a675) of yw eggs protein is known to be involved in vitelline membrane cross-linking ng at S8 . As moreng at S8 ; at S10Bng at S8 . Consistpxt eggs examined . In contrast to pxt mutants, however, defects in nurse cell dumping and egg elongation were not observed in follicles from females expressing pxt RNAi in the follicle cells. Thus the eggshell defects observed in pxt mutants are not simply a consequence of a block in nurse cell dumping or decreased egg elongation.Knocking down pxt expression causes similar changes in temporal gene expression to those observed in pxt mutant follicles pxt mutant follicles. The vm32e gene encodes a vitelline membrane protein that is expressed during stage 10. In pxt mutants, vm32e is more highly expressed during S10B and transcripts decline in abundance more slowly during stages 12\u201314 than is seen in wild-type follicles for 10 minor chorion proteins as determined by whole mount in situ hybridization . Additionally, the expression of many genes involved in follicle maturation are spatially regulated within the folliclar epithelium Our studies show that a large fraction of the genes involved in eggshell production can be identified by simply scoring for stage-specific changes in transcript levels during late oogenesis. Not only were virtually all of the previously known structural genes involved in producing yolk, the vitelline membrane and the chorion identified, but we also discovered at least 19 new candidate eggshell proteins , S2. Desdization . Many ofCG11381 (FBgn0029568), CG13083 (FBgn0032789), CG13084, CG13114, CG14796 (FBgn0025390), CG15570, CG4009, and CG31928. CG3074 (FBgn0034709) and CG13992 (FBgn0031756) are expressed at lower levels during stage 10 and may encode vitelline membrane proteins. The value of using temporal regulation as a criterion to identify eggshell components is highlighted by the fact that transcripts corresponding to the minor basement membrane components identified by Fakhouri et al. (2006) remain constant during eggshell formation . All three were scored as ovary-specific in FlyAtlas in situ hybridization to exhibit a posterior restriction Another class of predicted eggshell genes encode for proteases and protease inhibitors. Such proteins have recently been shown to be a major class of chicken eggshell components pxt, the COX-like enzyme, is expressed and pxd is absent throughout all stages of egg maturation. In contrast, the eggshell protein and putative peroxidase CG4009 is very highly expressed during S12 (The nature of the peroxidase that crosslinks the eggshell ring S12 . We propCG9747 (FBgn0039754) encodes for an acyl-CoA \u039411-desaturase, which is likely to desaturate palmitate; such enzymes are important for pheromone biosynthesis in other insects CG8303 (FBgn0034143) is highly expressed as stages 9/10A and 10B, and encodes for an acyl-CoA reductase, which activates fatty acids by adding CoA. Additionally two Elvol (elongation of very long chaing fatty acids) encoding genes, bond and CG2781 (FBgn0260942), are expressed at S10A and S10B, respectively. Bond has previously been shown to be required for both male and female fertility, and by in situ hybridization appears to be expressed during oogenesis in the main body follicle cells at stages 9 and 10 In addition to known and novel eggshell components, we identified many temporally regulated genes expressed during late follicle development. A number of putative lipid-processing genes exhibit stage specific expression, suggesting a role in yolk or pheromone production. pxt mutant follicles, but vitelline membrane genes continue to be expressed longer than normal. Some chorion genes turn on earlier than normal, while the expression of others is delayed or prolonged. Many possible mechanisms may underlie these changes. However, we are particularly interested in the possibility that Pxt coordinates the production of PGs that interact with other mechanisms to precisely control egg maturation.Pxt mutations partially uncouple morphological development and gene expression. Yolk protein genes turn off normally in In all sexually reproducing organisms the growth and development of the somatic and germ cells are mutually dependent and must be coordinated. Such coordination requires bi-directional communication. Historically, somatic cells were thought to regulate follicle development, including maintaining meiotic arrest, promoting meiotic resumption, and suppressing oocyte transcription prior to nuclear maturation There is emerging evidence that PG signaling coordinates germline and somatic development within mammalian follicles. While both oocyte and somatic maturation are delayed in COX2 knockout mice, it has been shown that the PGs are required in the soma for fertility 2alpha stimulates the release of oxytocin to mediate luteolysis or the regression of the corpus luteum 2alpha also play critical roles. Oxytocin initiates labor, inducing PGF2alpha, which maintains labor and dilates the cervix Female reproduction is regulated by a complement of hormones that are cyclically produced and secreted. One such hormone that interacts with PG signaling in mammals is oxytocin. Oxytocin plays critical roles in regulating the function of the corpus luteum, a transient endocrine organ that secretes hormones to regulate the menstrual cycle and the early stages of pregnancy. In the absence of pregnancy, PGF2 signals via cAMP and PKA to stimulate a promoter upstream of cyp19, leading to increased aromatase expression 2 secretion PGs and estrogen co-regulate each other in multiple cells types, including breast cancer cells. Breast tissue is the largest producer of estrogen in post-menopausal women; aromatase, Cyp19, leads to the production of estradiol. There is a high correlation between aromatase and COX2 expression in human breast cancer samples 2 and PGF2alpha are excessively produced in uterine and endometriotic tissues of women with endometriosis 2 stimulates the expression of all the steroidogenic genes needed to synthesis estradiol from cholesterol. This occurs via PGE2 activation of cAMP/PKA signaling which upregulates of the expression of steroidogenic acute regulatory gene (StAR) and cyp192 signaling leads to SF1 out competing other transcription factors, Chicken Ovalbumin Upstream Promoter Transcription Factor (COUP-TF) and Wilms' tumor-1 (WT-1), for binding to steroidogenic gene promoters PGs and estrogen also interact in endometriotic tissue. Both PGEOur results encourage future efforts to further establish the roles for PG signaling during Drosophila egg maturation and specifically, to learn how PGs are connected to steroid hormones. The Drosophila hormone ecdysone plays several critical roles during oogenesis. The loss of ecdysone signaling arrests follicle development at stage 8 Figure S1Temporal control and tissue-specificity of follicle gene expression. (A\u2013B) The temporal expression patterns of 5 eggshell protein transcripts (A) and 5 non-structural transcripts (B) determined by RT-PCR. Their times of expression agree closely with the microarray data. (C\u2013D) The tissue-specificity of the genes in A\u2013B were assayed by RT-PCR using RNAs from the six indicated sources (1\u20136). Rps17 served as a loading control for all panels in C\u2013D. In agreement with our RT-PCR tests, the FlyAtlas expression project found that 18 of 19 candidate eggshell genes were ovary-specific. Most of the temporally regulated non-structural genes were expressed more widely.(TIFF)Click here for additional data file.Figure S2Effect of pxt mutation on the expression of selected non-egghell genes. RT-PCR was used to assay transcript levels from the indicated genes during the stages 10B, 12 or 14 from wild type (y w) or pxtf01000 females (pxt\u2212/\u2212). Rps17 transcripts served as a loading control. pax\u200a=\u200aFBgn0041789; gstd1\u200a=\u200aFBgn0001149; cg30382\u200a=\u200aFBgn0050382; ste24a\u200a=\u200aFBgn0034176; cyp18a1\u200a=\u200aFBgn0010383; cg30334\u200a=\u200aFBgn0050334; yp3\u200a=\u200aFBgn0004047; cyp4p2\u200a=\u200aFBgn0033395; cg10013\u200a=\u200aFBgn0038012; cg13937\u200a=\u200aFBgn0035287; cg8027\u200a=\u200aFBgn0033392; cyp4s3\u200a=\u200aFBgn0030615; cg4066\u200a=\u200aFBgn0038011, cg1681\u200a=\u200aFBgn0030484.(TIFF)Click here for additional data file.Table S1Expression of the well-characterized eggshell protein genes. Table of transcript levels and stage-specifity as determined by microarray for the 30 well-characterized eggshell protein genes, and for three yolk protein genes. The previously characterized temporal expression (\u201cRef\u201d) is listed under \u201cPattern.\u201d References: 1Burke et al. (1987) Dev Biol 124: 441\u2013450. 2Claycomb et al. (2004) Dev Cell 6: 145\u2013155. 3Fakhouri et al. (2006) Dev Biol 293: 127\u2013141. 4Parks et al. (1986) Dev Biol 117: 294\u2013305. 5Parks and Spradling. (1987) Genes Dev 1: 497\u2013509. 6Popodi et al. (1988) Dev Biol 127: 248\u2013256. 7Yakoby et al. (2008) Dev Cell 15: 725\u2013737.(DOCX)Click here for additional data file.Table S2Putative new eggshell protein genes. Table of transcript levels and stage-specifity as determined by microarray for the putative eggshell protein genes. 1Corresponds to Affy; gene model uncertain. 2See also Yakoby et al. 2008.(DOCX)Click here for additional data file.Table S3Other genes regulated in late follicles by category. Table of transcript levels and stage-specifity as determined by microarray for genes expressed temporally during late follicle development. 1See also Yakoby et al. 2008.(DOCX)Click here for additional data file."} +{"text": "This study was conducted to identify the most common sensitizing food and inhalant allergens in physician-diagnosed allergic children and adults in North India.\u00ae Phadiatop and fx5 , a technology considered as the gold standard for IgE antibody blood testing worldwide. For identification of the sensitizing allergens atopic patients were further tested by ImmunoCAP\u00ae Specific IgE using a broad panel of common Indian allergens covering 17 foods and 19 inhalants (singles/mixes). Total IgE level was also determined for each atopic patient.274 allergy-diagnosed patients, divided into Group A (aged 6\u201312 yrs) and Group B (aged 12\u201365 yrs), were enrolled in the study. They were classified as atopic if had at least one positivity when screened with ImmunoCAPD. farinae (83%) was the most prevalent sensitizing allergen followed by cockroach (79%), weed pollens (29-50%), tree pollen (16-29%), grass pollen mix (26%) and mold mix (25%). Less than 20% of patients tested positive to cow\u2019s milk, tomato, spinach, aubergine, soybean, egg white, dog dander, legume mix, chicken, cat dander and fish. The geometric mean of total IgE was 943 kU/l .Phadiatop/fx5 determined 59% (162/274) of the patients as atopic, where of 159 were included in further evaluation; 10% were in Group A and 90% were in Group B. Higher proportion (36%) of patients had the medical history of urticaria followed by atopic dermatitis (26%), asthma (23%) and rhinitis (23%). The commonest sensitizing food allergen was banana (68%) followed by sesame seeds (66%), lemon (45%), rice (31%), wheat (24%), cashew (23%) and peanut (21%). Among inhalants, house dust mite, \u00ae which provided useful native information of prevalent sensitizing Indian allergens that would improve cost effectiveness of allergy treatment and hence increase the quality of life of allergic patients in India. Phadiatop/fx5 revealed that the physicians\u2019 diagnosis of IgE mediated allergy was accurate only in 59% of cases and thus highlights the importance of using allergy tests in conjunction with clinical findings for correct allergy diagnosis.This is the first Indian sensitization data of this dignity analyzed by ImmunoCAP"} +{"text": "Receptor activator of nuclear factor-\u03baB ligand (RANKL), a member of tumor necrosis factor (TNF)-\u03b1, is produced by osteoblasts (Obs) and stimulates its receptor RANK on osteoclast (Oc) progenitors to differentiate them to osteoclasts. WP9QY peptide designed to mimics TNF receptor's contact site to TNF-\u03b1 was known to abrogate osteoclastogenesis in vitro by blocking RANKL-RANK signaling. WP9QY ameliorated collagen-induced arthritis and osteoporosis in mouse models. Here we report that the peptide surprisingly exhibited bone anabolic effect in vitro and in vivo.WP9QY was administered subcutaneously to mice three times per day for 5 days at a dose of 10 mg/kg in normal mice, followed by peripheral quantitative computed tomography (pQCT) and histomorphometrical analyses. To clarify the mechanism by which the peptide exerted the bone anabolic effect, we examined the effects of the peptide on osteoblast (Ob) differentiation/mineralization with mouse MC3T3-E1 (E1) cells and human mesenchymal stem (MSC) cells, and those on osteoclast (Oc) differentiation with RAW264 cells in the presence of sRANKL.WP9QY augmented bone mineral density (BMD) significantly in cortical bone not in trabecular bone. Histomorphometrical analysis showed that the peptide had little effect on osteoclasts in distal femoral metaphysis, but markedly increased bone formation rate in femoral diaphysis. The peptide markedly increased alkaline phosphatase activity in E1 and MSC cell cultures and decreased tartrate-resistant acid phosphatase activity in RAW264 cell culture in a dose-dependent manner, respectively. In addition, the peptide stimulated mineralization evaluated by alizarin red staining in E1 and MSC cell cultures. The anabolic effect of WP9QY peptide was enhanced markedly by addition of BMP2.Increases in mRNA expression of IGF1, collagen type I, and osteocalcin were observed in E1 cells treated with the peptide for 12 and 96 h in GeneChip analysis. Addition of p38 MAP kinase inhibitor reduced ALP activity in E1 cells treated with the peptide, suggesting a signal through p38 was involved in the mechanisms.Taken together, the peptide abrogated osteoclastogenesis by blocking RANKL-RANK signaling and stimulated Ob differentiation/mineralization with unknown mechanism in vitro. However, in our experimental conditions the peptide exhibited bone anabolic effect dominantly in vivo. Since the peptide is known to bind RANKL, we hypothesize that the peptide shows the bone anabolic activity with reverse signaling through RANKL on Obs."} +{"text": "The overuse or misuse of antibiotics has accelerated antibiotic resistance, creating a major challenge for the public health in the world. Sewage treatment plants (STPs) are considered as important reservoirs for antibiotic resistance genes (ARGs) and activated sludge characterized with high microbial density and diversity facilitates ARG horizontal gene transfer (HGT) via mobile genetic elements (MGEs). However, little is known regarding the pool of ARGs and MGEs in sludge microbiome. In this study, the transposon aided capture (TRACA) system was employed to isolate novel plasmids from activated sludge of one STP in Hong Kong, China. We also used Illumina Hiseq 2000 high-throughput sequencing and metagenomics analysis to investigate the plasmid metagenome. Two novel plasmids were acquired from the sludge microbiome by using TRACA system and one novel plasmid was identified through metagenomics analysis. Our results revealed high levels of various ARGs as well as MGEs for HGT, including integrons, transposons and plasmids. The application of the TRACA system to isolate novel plasmids from the environmental metagenome, coupled with subsequent high-throughput sequencing and metagenomic analysis, highlighted the prevalence of ARGs and MGEs in microbial community of STPs. The wide use of antibiotics results in environmental releases, accelerating antibiotic resistance development in hospital wastewater, domestic sewage and livestock manure. Sewage receives the gut bacteria previously exposed to antibiotics, so that sewage treatment plants (STPs) are considered as important reservoirs for antibiotic resistance genes (ARGs) Activated sludge and biofilm in STPs are characterized with high microbial density and diversity which may facilitate ARG horizontal transfer In this study, we employed the transposon aided capture (TRACA) system tetG increased from 1.1\u00d7105 copies/ng of total DNA to 4.1\u00d7106 copies/ng of plasmid DNA after being purified by QIAGEN Plasmid Purification Kit and treated by Plasmid Safe DNase . However, PCRs demonstrated that 16S rRNA gene still occurred in the plasmid DNA samples, showing that chromosomal DNA contamination was not completely removed.In order to deeply explore the plasmid metagenome, we employed a plasmid purification kit and a Plasmid Safe DNase to concentrate the plasmids in the DNA samples. Quantitative real time PCR (qPCR) indicated that the level of tetracycline resistance gene type fe DNase , indicatActinobacteria, Chloroflexi, Proteobacteria, Bacteroidetes, and Firmicutes aromatic compound degradation or heavy metal resistance determinants Ralstonia solanacearumPlasmids are considered as the main vehicle for intercellular horizontal transfer of genetic materials and we therefore searched the metagenome for plasmid-associated DNA. Mapping all the reads to NCBI Plasmid Genome Database revealed that the number of matched reads ranged from 4,588 to 122,427 under different hit lengths (50\u2013100 bp) and similarities (90%\u2013100%) . In thisbacteria , which wbacteria . Among tAeromonas hydrophila plasmid pRA3 (Accession No. DQ401103.1) and Aeromonas punctata plasmid pFBAOT6 (Accession No. CR376602.1). Plasmid pST1 contained a tetracycline resistance gene (tetB(P)), an aminoglycoside resistance gene (aadA5) and a transposon (ISSm2) (Sm2 was similar to ISSm1 in terms of DNA sequence aadA5The sequencing reads of one contig was well assembled to generate a previously undescribed plasmid (pST1), judging from a 26-bp overlap on both end of the contig sequence. BLAST searches demonstrate that the overlap fragment is completely identical to the gene encoding replication initiation protein on (ISSm2) . The hypIn order to acquire more novel plasmids, the TRACA system was used to isolate plasmids from extracted DNA of the activated sludge. Two plasmids (pST2 and pST10) were acquired and their sizes were 3,206 bp (pST2) and 3,038 bp (pST10), respectively . Genes aampC encoding \u03b2-lactamase has been detected in the microbial isolates from wastewater, surface water, and even drinking water films Pps1) and Acidovorax avenae (accession number: AF086815).For plasmid pST2, the only known putative function encoded by this plasmid was \u03b2-lactam resistance . The genms Pps1) that is tet) often occur in STPs with high abundances tet genes, tetB(P), tetM and tetG had the highest abundance were relatively more abundant in this STP macB was the most abundant ARG (175 reads) consisting of 23,137 sequences of 380 ARGs encoding resistance to 249 antibiotics bundance . However5 reads) , which e5 reads) . The macampC, VEB-3bla, VIM-2bla and IMP-1bla), aminoglycoside , sulphonamides (sulI), trimethoprim (dfrA1) and quaternary ammonium compounds (qacE\u03941). Alignment against the transposon and insertion sequence database ISfinder Pps1 had the highest abundance, followed by ISSm2 (679 reads) and ISAba3 (306 reads). This was further confirmed by the presence of ISPps1 on plasmid pST10 and ISSm2 on plasmid pST1 since the two high-copy plasmids were isolated from the activated sludge.The mobility of ARGs depends on the MGEs, such as plasmids, integrons and transposons pA is often carried by pathogen Shigella flexneri associated with multiple antibiotic resistances S. flexneri) also had high abundance, which usually includes \u03b2-lactamase (OXA-30bla) and streptomycin adenyltransferase (aadA1) genes in their gene cassettes in silico analysis methods of metagenomics data.Pathogenic bacteria are able to induce different diseases by expression of different combinations of VFs, which is often mediated by MGEs Our results show that various MGEs are present in STPs and these elements may play important roles in horizontal transfer of various ARGs among the bacterial species. This highlights the fact that STPs serve as potential recruitment pools for antibiotic resistant bacteria. ARGs may enter into water or soil environments along with STP effluent and the agricultural application of sludge tetG (tetracycline resistance gene) in the total DNA directly isolated from sludge and the concentrated plasmid metagenome DNA. The copy number was normalized to DNA mass and the qPCR procedures were previously described in detail Activated sludge samples were collected from Shatin Sewage Treatment Plant at three times, separately on August 17, October 12 and November 18 of 2010. Details about the STP were previously described Tn5TM Insertion Kit . Each reaction was composed of Plasmid Safe digested metagenomic DNA (0.5 \u00b5g), transposase (1 U), 10\u00d7 EZ-Tn5 reaction buffer (2 \u00b5l) and brought to a final volume of 20 \u00b5l by adding sterile pure water. Reactions were conducted for 2 h at 37\u00b0C and stopped by adding EZ-Tn5 stop buffer (2 \u00b5l) and heating at 70\u00b0C for 10 min. Then, reaction mixtures were applied to transform TransforMax\u2122 EPI300\u2122 Chemically Competent E. coli according to the manufacturer's introduction. Transformants were selected on LB agar supplemented with 50 mg/l kanamycin and cultured in Growth Media to amplify low-copy plasmid clones. Plasmids with transposon insert were extracted and purified using MiniBEST Plasmid Purification Kit . The plasmids acquired were sequenced by BGI using the primer walking strategy.Plasmids were isolated from Plasmid Safe treated metagenomic preparations using the TRACA system according to Jones et al. High-throughput sequencing was performed by BGI using Illumina Hiseq 2000. The sequencing strategy was index PE101+8+101 cycle . More than 1 Gb of sequences was generated for the sludge DNA sample. The raw reads containing three or more \u201cN\u201d or contaminated with adaptors were removed and the remaining clean reads (98.9%) were used for further analysis. The metagenomic data have been deposited at NCBI Sequence Read Archive under accession number SRP007256.1. Nucleotide sequences of the plasmids were deposited at GenBank under accession numbers JN098513 - JN098515.\u221210). MetaGene program \u221210).All sequencing reads were assembled by using SOAPdenovo with Kmer at 55. The contigs were compared against the non-redundant protein database at NCBI GenBank using BLAST .In order to characterize the plasmid metagenome, all reads were aligned to plasmid genome sequences available in the NCBI RefSeq database . The alignment was performed using BLAST against the known plasmids. BLAST hits (blastn) were determined for the alignments with a nucleotide sequence identity above 95% over a length of at least 90 bp. Plasmid coverage of pST1, pST2 and pST10 was calculated by counting the number of reads aligned at each base according to Kristiansson et al. A database of resistances genes was created from all sequences in ARDB DNA sequences of virulence factors (VFs) were downloaded from VFDB Table S1Assembling analysis of high-throughput sequencing reads in plasmid metagenome of activated sludge with the optimal Kmer at 55.(DOC)Click here for additional data file.Table S2Prediction of open reading frames (ORFs) using Metagene Annotator Software.(DOC)Click here for additional data file.Table S3Statistical analysis on functional classification and species annotation of the available open reading frames according to different databases.(DOC)Click here for additional data file.Table S4Matched assembled contigs of plasmids in the activated sludge of Shatin STP.(DOC)Click here for additional data file.Table S5Matched high-throughput sequencing reads of plasmids in the activated sludge of Shatin STP.(DOC)Click here for additional data file.Table S6Matched high-throughput sequencing reads of antibiotic resistance genes (ARGs) in the activated sludge against Antibiotic Resistance Database (ARDB).(DOC)Click here for additional data file.Table S7Matched high-throughput sequencing reads of integrons and gene cassettes in the activated sludge of Shatin STP.(DOC)Click here for additional data file.Table S8Matched high-throughput sequencing reads of insertion sequence common region transposases in plasmid metagenome of activated sludge of Shatin STP.(DOC)Click here for additional data file.Table S9Matched high-throughput sequencing reads of virulence factors in plasmid metagenome of activated sludge of Shatin STP.(DOC)Click here for additional data file.Figure S1Calibration curves of quantitative real-time PCR for tetracycline resistance gene tetG generated with serial dilutions of plasmid vector carrying the target gene tetG (A) and comparison of tetG abundance in the environmentally total DNA and the concentrated plasmid metagenome isolated from activated sludge (B).(TIF)Click here for additional data file.Figure S2\u221210).KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway classification of plasmid metagenome of the activated sludge in Shatin STP Click here for additional data file.Figure S3Functional classification of plasmid metagenome in the activated sludge of Shatin STP against evolutionary genealogy of genes: Non-supervised Orthologous Groups (eggNOG) databases .(TIF)Click here for additional data file.Figure S4\u221210).Species annotation of ORFs of plasmid metagenome in the activated sludge of Shatin STP against the non-redundant protein database at NCBI GenBank Click here for additional data file.Figure S5Number of matched high-throughput sequencing reads against NCBI plasmid genome database by using different cut-offs (hit length and sequence identity). The number of hits has significant negative correlation with both hit length and sequence identity.(TIF)Click here for additional data file."} +{"text": "The Co2+ cations are linked by means of bridging ligands into polymeric chains along [010]. These chains are further connected to each other through hydrogen bonds involving partially occupied disordered water mol\u00adecules; thus, sheets parallel to (001) are formed. One of the positions of disordered water O atom lies on a twofold axis. Two atoms of the aliphatic chain of one of the butanoate anions are disordered over two positions each. In the title coordination polymer, {[Co(C DOI: 10.1107/S1600536811019271/ya2137Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Quantitative trait loci (QTLs) for bone mineral density (BMD) are defined as regions of the genome that contain sequence variations that cause differences in either bone deposition or rates of resorption. In this study, we investigate QTLs for BMD of whole bones using femurs and tibiae from the BXD family of recombinant inbred (RI) strains derived by crossing C57BL/6J (B6) and DBA/2J (D2) inbred strains.http://www.genenetwork.org). Candidate genes in QTL regions were ranked using PGMapper. SNP genotypes of candidate genes were verified directly using PCR amplification and sequencing.We studied femurs and tibiae from a total of 46 strains at 3 months-of-age. Bones were quantified using the PIXImus dual-energy X-ray absorptiometer (DXA) system. QTL mapping was carried out using simple and composite interval mapping in GeneNetwork (Our data show:1) A high correlation between BMD of the femur and tibia across the panel of BXD strains;2) A high correlation between BMD of femur and tibia within sex and a moderate positive correlation between sexes;3) A QTL on chromosome 15 for the BMD in femur and tibia in male mice located in a 10 Mb region between 42 and 52 Mb;Trps1, Ext1, and Slc30a8.4) A total of 48 transcripts within the Chr 15 QTL or which three are particularly attractive candidate genes\u2014We have identified QTLs for BMD using a set of 46 BXD RI strains. Further investigation of the three candidate genes located in this QTL on chromosome 15 is essential. Despite limitations, PIXImus is a valuable tool for studying BMD and skeletal development of small animals."} +{"text": "Guidelines therapy for allergic rhinitis (AR) recommends new-generation H1-antihistamines and intranasal corticosteroids as main treatment. The objective for this study was to evaluate the effects of intranasal therapy with azelastine hydrochloride and budesonide (isolated and combined) using nasal provocation test (NPT) and acoustic rhinometry in patients with AR.The study population consisted of 28 patients with diagnostic of persistent AR (ARIA consensus). This was a randomized, crossover study. Subjects were randomly assigned to receive either azelastine hydrochloride (140 mcg/nostril) or budesonide (64mcg/nostril) or both drugs. All patients received the three treatments using nasal spray twice daily, each period of treatment lasted 30 days and washout period was 7 days. Subjects were submitted to nasal provocation test (NPT) with histamine before and after each period of treatment. Nasal responsiveness to histamine was monitored based on subjective (symptom score) and objective parameters (acoustic rhinometry) to compare the treatments. After acoustic rhinometry measure (baseline) histamine was instilled in nasal cavity (0.5mg/mL/nostril) via nasal spray. Minimal cross-area (MCA2) was measured by acoustic rhinometry at times 1; 4; 8; and 12 minutes after NPT for each histamine concentration up to positive response occurs . The criteria for a positive response were histamine dose and rhinometric measure time causing at least 20% fall in MCA2 (NPT20). NPT was stopped when a positive response occurred. Statistical significance was assessed by paired t-test.Baseline MCA2 decreases significantly after azelastine therapy (p=0.01) and did not change with budesonide. Combination therapy demonstrated significantly increasing in baseline MCA2, namely improvement of nasal patency (p=0.005). MCA2 after NPT increased with budesonide (p=0.02) and combined therapy (p=0.002), demonstrating the protective effect of therapy. There was a decrease in MCA2 NPT20 after azelastine hydrochloride therapy (p=0.02). Score symptoms during NPT decreases significantly after all treatments .Azelastine hydrochloride therapy combined with budesonide might provide more therapeutic benefits than isolated drugs in nasal patency and symptoms score in patients with AR."} +{"text": "The European Neolithization \u223c6000\u22124000 BC represents a pivotal change in human history when farming spread and the mobile style of life of the hunter-foragers was superseded by the agrarian culture. Permanent settlement structures and agricultural production systems required fundamental innovations in technology, subsistence, and resource utilization. Motivation, course, and timing of this transformation, however, remain debatable. Here we present annually resolved and absolutely dated dendroarchaeological information from four wooden water wells of the early Neolithic period that were excavated in Eastern Germany. A total of 151 oak timbers preserved in a waterlogged environment were dated between 5469 and 5098 BC and reveal unexpectedly refined carpentry skills. The recently discovered water wells enable for the first time a detailed insight into the earliest wood architecture and display the technological capabilities of humans \u223c7000 years ago. The timbered well constructions made of old oak trees feature an unopened tree-ring archive from which annually resolved and absolutely dated environmental data can be culled. Our results question the principle of continuous evolutionary development in prehistoric technology, and contradict the common belief that metal was necessary for complex timber constructions. Early Neolithic craftsmanship now suggests that the first farmers were also the first carpenters. After the last Ice Age \u223c12,000 BP, the Central European landscape changed from steppes to dense woodlands A precise chronological framework beyond radiocarbon dates and LBK pottery typology is required for a deeper understanding of the Neolithization process Quercus spp.) timbers from four water well constructions excavated in Altscherbitz, Brodau and Eythra and a tube-like well lining (using hollowed trunk sections). The chest-like construction in well B served to stabilize the construction pit before a hollowed trunk was inserted . Well E2Well A was discovered at the margin of an LBK settlement of nearly 100 typical longhouses and a cemetery of approximately two dozen graves . From thHowever, re-used timbers excavated from the surrounding pit were more than 100 years older than the well lining itself. This widespread dating evidence from well A implies a long settlement activity of at least four generations preceding the construction of the well. Our dendrochronological dating allows the determination of an ascertained time span for a particular early Neolithic settlement.Triticum monococcum) and emmer (Triticum dicoccum) (Pisum sativum) and lentils (Lens culinaris). Oils were obtained from linseed (Linum usitatissimum) and poppy (Papaver somniferum). Wild fruits supplemented the diet, and included strawberries, sloe, apples, raspberries and hazelnuts. Two plants that have been considered archaeophytes in Central Europe were found in abundance: the bladder cherry and the black henbane (Hyoscyamus niger). Henbane is a strong hallucinogenic drug and potential medicine. Its utilization as a medicinal plant or ritual drug has been suggested elsewhere Rich botanical remains from the well fill provide insight into past environmental conditions and the early Neolithic diet. The staple food consisted of two types of hulled wheat, einkorn (icoccum) . CarbohyThe lower part of well A, which was filled with sediment after its abandonment, contained over 25 complete LBK vessels as well Many of the tool marks on the timber surfaces can be attributed to typical early Neolithic ground stone adzes. Unlike later archaeological cultures, parallel-hafted axes were unknown in the LBK. The predominant tool for woodworking was the transversely hafted adze with the ground stone blades that are extensively known from the archaeological record Cerambyx cerdo L.) in 27% of the analyzed well timbers Click here for additional data file.Figure S1Archaeological plan of the LBK settlement from Altscherbitz with the located water well, nearly 100 typical longhouses and a cemetery of about two dozen graves.(PDF)Click here for additional data file.Figure S270-ton block with the Altscherbitz well encased.(PDF)Click here for additional data file.Figure S3Indoor excavation of the Altscherbitz well.(PDF)Click here for additional data file.Figure S43D laser rendering of the Altscherbitz basal frame.(PDF)Click here for additional data file.Figure S5Timber from the Altscherbitz well lining and sawn cross section sample.(PDF)Click here for additional data file.Figure S6Close-up view of a cross section . The last two or three heartwood rings are discolored.(PDF)Click here for additional data file.Figure S7Great capricorn beetle galleries (Cerambyx cerdo L.).(PDF)Click here for additional data file.Figure S842 tree-ring series from split timbers from the Altscherbitz well lining can be attributed to one individual tree because of their similarity.(PDF)Click here for additional data file.Figure S9Split timbers from the construction pit can be attributed to one individual tree trunk.(PDF)Click here for additional data file.Figure S1037 trees reconstructed from 147 Altscherbitz timber tree-ring series. Sapwood: blackened; waney edge: red; pith: black dot.(PDF)Click here for additional data file.Figure S11Relationship between average growth rate (AGR) and mean segment length (MSL) of the Altscherbitz dataset.(PDF)Click here for additional data file.Figure S12Smoothed regional curves representing the average age trend of recent oaks from Central Eastern Germany (green) and Early Neolithic oaks from the Altscherbitz well construction (red).(PDF)Click here for additional data file.Figure S13Synchronization of the 124 Altscherbitz tree-ring series. (A) EPS over 50 years, lagged by 25 years, (B) replication, (C) individual tree-ring series (black) in overlap with mean (red), (D) mean chronology in overlap with the chronology from the Main river valley after 10-year smoothing.(PDF)Click here for additional data file.Figure S14Well from Brodau in the course of excavation with a piglet in the construction pit.(PDF)Click here for additional data file.Figure S15Highly decomposed oak timber from the Brodau well.(PDF)Click here for additional data file.Figure S16A) Brodau tree-ring series in overlap. (B) Brodau mean chronology (red) in overlap with the Altscherbitz reference chronology (blue).((PDF)Click here for additional data file.Figure S17Joining techniques of early Neolithic well constructions.(PDF)Click here for additional data file.Figure S18A) 18 tree-ring series from Eythra well E1 in overlap. (B) Mean chronology from E1 (red) in overlap with the Altscherbitz reference chronology (blue).((PDF)Click here for additional data file.Figure S19Eythra well E2: sketch of timber remains from structures 21 and 22.(PDF)Click here for additional data file.Figure S20A) Tree-ring series from the Eythra well E2 in overlap (grey). (B) The mean chronology (red) of E2 dated against the Altscherbitz mean (blue) chronology.((PDF)Click here for additional data file.Figure S21Environmental change in the Early Neolithic. (A) Pollen-based European temperature reconstruction, (B) subfossil-based Alpine treeline reconstruction, (C) temporal distribution of glacial 95 wood remains, and (D) peat bog-based hydroclimatic reconstruction from the UK.(PDF)Click here for additional data file.Table S1Grid report of the correlation results between chronologies from well A, B, E1, E2 and the Main river valley. TBP\u200a=\u200at-value after Baillie and Pilcher, THO\u200a=\u200at-value after Hollstein, Gl\u200a=\u200a% of Gleichl\u00e4ufigkeit, r\u200a=\u200acorrelation coefficient.(PDF)Click here for additional data file.Table S2Tree-ring inventory.(PDF)Click here for additional data file.Data S1Tree-ring width data (Tucson Format).(PDF)Click here for additional data file."} +{"text": "Sox10, drives the survival and differentiation of the melanocyte lineage. However, the role that Sox10 plays in postnatal melanocytes is not established. We show in vivo that melanocyte stem cells (McSCs) and more differentiated melanocytes express SOX10 but that McSCs remain undifferentiated. Sox10 knockout Sox10) results in loss of both McSCs and differentiated melanocytes, while overexpression of Sox10 (Tg(DctSox10)) causes premature differentiation and loss of McSCs, leading to hair graying. This suggests that levels of SOX10 are key to normal McSC function and Sox10 must be downregulated for McSC establishment and maintenance. We examined whether the mechanism of Tg(DctSox10) hair graying is through increased expression of Mitf, a target of SOX10, by asking if haploinsufficiency for Mitf (vga9Mitf) can rescue hair graying in Tg(DctSox10) animals. Surprisingly, vga9Mitf does not mitigate but exacerbates Tg(DctSox10) hair graying suggesting that MITF participates in the negative regulation of Sox10 in McSCs. These observations demonstrate that while SOX10 is necessary to maintain the postnatal melanocyte lineage it is simultaneously prevented from driving differentiation in the McSCs. This data illustrates how tissue-specific stem cells can arise from lineage-specified precursors through the regulation of the very transcription factors important in defining that lineage.During embryogenesis, the transcription factor, Sox10, in the melanocyte lineage results in loss of McSCs and consequently leads to premature hair graying. Through the use of mouse transgenics, we demonstrate that by changing Sox10 levels, melanocytes of the hair can preserve their ability to survive and produce normally pigmented hairs while also allowing a portion of them to fulfill the role of an undifferentiated McSC. We also discovered that Mitf, a downstream target of SOX10 and the master regulatory transcription factor for melanogenesis, appears to participate in McSC maintenance, perhaps by the negative regulation of SOX10-dependent processes. These observations raise the idea that adult stem cells, like McSCs, may rely on cell type specific transcription factors for their specification and survival, but that these transcriptions factors also have to be carefully regulated to maintain a stem cell fate.The melanocyte stem cells (McSCs) that reside in the hair follicle are critical for generating the melanocytes that will differentiate and produce pigment for the hair during successive rounds of hair growth. The inappropriate maintenance of McSCs results in hair graying. Thus, our understanding of McSC biology is enhanced through the study of hair graying mouse models. We have discovered that sustained expression of the transcription factor, In the adult animal, tissue-specific stem cells exist in a number of organs and function to sustain these tissues during normal homeostasis. However, our understanding of the origin and establishment of tissue-specific stem cells during organogenesis is incomplete. Using melanocytes as a model, we investigated the process of lineage-specific stem cell fate acquisition by examining the role of the transcription factor SOX10 in the formation of the melanocyte stem cell (McSC) within the mouse hair follicle.Melanocytes of the hair follicle have gained increasing attention for studying cell-specific contributions to organ development and maintenance. Individual hair follicles act as \u2018mini-organs\u2019 The most critical time point for establishing McSCs appears to be during hair morphogenesis. Studies using the KIT-blocking antibody, ACK, to deplete melanocyte populations perinatally show that McSCs inhabit hair follicles around P4, demonstrated by the fact that they survive independent of KIT-signaling and are sufficient to restore coat color pigmentation Sox10, where it is expressed in melanoblasts of the skin and melanocytes of the hair bulb but absent from McSCs, suggests that transcriptionally downregulating Sox10 is the mechanism by which melanoblasts acquire a McSC fate. This hypothesis fits well with the known function of Sox10 as a transcription factor that participates in melanocyte differentiation by upregulating Mitf, the master regulatory gene for melanogenesis. The loss of melanin synthesis proteins, TRP1 and TYR, within presumptive McSCs further supports this idea since SOX10 transcriptionally activates these genes, and that TYR is required by mouse melanocytes to generate pigment Sox10 is expressed during neural crest development and its loss embryonically results in several neurocristopathies, including congenital hypopigmentation Sox10 null mice has precluded functional analysis of Sox10 in adult melanocytes. Using conditional transgenics we can now explore the role of Sox10 postnatally in the melanocytes of the mouse hair follicle.The subpopulation-specific expression of the transcription factor Sox10 for normal hair pigmentation. However, constitutive expression of Sox10 by McSCs disrupts their maintenance by driving their premature differentiation. We also demonstrate that Mitf contributes to this regulation, likely through a negative feedback mechanism. Together, these data support the theory that transcription factors responsible for the specification of lineage-defined precursors can later participate in the specification and maintenance of stem cells derived from those precursors.Here we report that postnatal mouse melanocytes both express and require Tyr::CreERT2, can target cells with these properties when induced either during postnatal development or within adults tm1sorTyr::CreERT2; Rosa26 pups were given a pulse of tamoxifen (TAM) on postnatal days 2 and 3 (P2\u20133), and assessed for recombined cells by \u03b2-galactosidase staining. Melanocytes in Tyr::CreERT2 mice express CRE at P2, and within the same hair cycle as TAM treatment (P14), we observed LacZ+ cells in the hair bulge and bulb we observed the retention of LacZ+ cells in the hair bulge and LacZ+ progeny in the hair bulb . In this study, we define the McSC population by several characteristics: cells that exist within the hair bulge, are capable of self-renewal, and can give rise to melanocyte progenitors that colonize the newly developing hair bulb. Previously, we have shown that the transgenic line, and bulb \u2013B. This air bulb . This colabeling and confSox10 reporter mice + cells within hairs co-express SOX10 regardless of time point, location or differentiation status of the hair (hair bulge) and the upper transitory portion (UTP) of the hair since these regions contained the majority of DCT+ melanocytes that exist along the hair shaft (+/DCT+ cells comprise 87% of the LPP+UTP melanocytes at anagen (7dpp), and 96% of bulge+SHG melanocytes at catagen corresponds with a dramatic escalation of the percentage KIThigh cells and a moderate increase of TRP1+ cells amongst LPP+UTP melanocytes. In contrast, entry into catagen (21dpp) is associated with LPP+UTP melanocytes downregulating MITF while still retaining TRP1, KIT and SOX10.In contrast to SOX10, the expression patterns of other melanocyte markers, MITF, KIT, TRP1 and TYR, within postnatal hairs is more variable , S5, S6.ectively . As hair+ melanocytes within the hair bulb also double-label with SOX10, MITF, TRP1 and TYR , DCT and TYR , S4, S5. and TYR .Sox10 influences these expression patterns within follicular melanocytes.This analysis demonstrates that while the differentiation status of melanocytes that exists within the hair bulge fluctuates in concert with hair morphogenesis and adult hair cycling, SOX10 expression remains static amongst LPP and UTP melanocytes. We next assessed how perturbation of Sox10-dependent Sox10 plays an important role in postnatal melanocyte biology. We tested this by generating fl/fl; Tyr::CreERT2Sox10 mice to conditionally knockout Sox10 in mouse melanocytes postnatally Tyr::CreERT2 is effective at inducing recombination of floxed alleles in a significant number of McSCs as well as the more differentiated melanocytes when TAM is administered transiently during perinatal growth or during adult anagen fl/fl; Tyr::CreERT2Sox10 mice in regions of the skin where newly grown hairs were emerging. This loss of pigmentation was not observed in similarly-treated fl/flSox10 and fl/+;Tyr::CreERT2Sox10 mice or fl/fl; Tyr::CreERT2Sox10 mice that were not treated with tamoxifen . Nevertheless, these data demonstrate that the hypopigmentation observed with Sox10 knockout is due to an overall loss of bulb melanocytes and a deficiency in their ability to produce pigment.In light of our discovery that SOX10 is retained by both McSCs and differentiated bulb melanocytes within the hair follicle, paired with previous in vitro experiments indicating that TYR expression in mouse is amoxifen \u2013B. Usingamoxifen . This inentation . This wa2 adults . This ingulation \u2013D. The fT2Tyr::CreER transgene is effective at inducing recombination in McSCs Sox10 knockout on LPP (bulge) melanocytes. Using KIT as our marker for melanocytes, we discovered that LPP melanocytes are decreased in fl/fl; Tyr::Sox10CreERT2 mice when induced perinatally or as adults /+ animals compared to wild type and hair graying (Tg(DctSox10)/+ mice exhibit hypopigmentation that is evident as small, ventral belly spots that are highly penetrant /Tg(DctSox10) mice at P8 have more extensive hypopigmentation with large white ventral spots that encompass the majority of the belly, dorsal spotting and occasional head spots. The white spotting observed with Tg(DctSox10) suggests that overexpression of Sox10 affects the embryonic melanoblast population. Tg(DctSox10)/Tg(DctSox10) mice also display variable loss of hair pigmentation (premature hair graying) after the onset of the first adult hair cycle /Tg(DctSox10) mice continues to increase progressively as these animals age (not shown). Tg(DctSox10)/+ mice also exhibit hair graying but with a reduction in severity and with a later onset, beginning after the second adult hair cycle . Hair graying in the Tg(DctSox10) line was first examined histologically in mice after the first adult hair cycle (between 9\u201322 weeks in age) and after hair cycle synchronization by plucking (Tg(DctSox10)/+ mice, the majority of hairs were both pigmented and contained LPP melanocytes . In contrast, Tg(DctSox10)/Tg(DctSox10) mice exhibited primarily non-pigmented hairs that lacked LPP melanocytes (75.6\u00b14.3%). From these observations we conclude that Tg(DctSox10)-induced hair graying is a direct consequence of McSC deficiency.Hair graying in plucking . AnalysiTg(DctSox10)/Tg(DctSox10) animals exhibit premature hair graying at the first adult hair cycle suggests that the loss of McSCs observed in these animals occurs during hair morphogenesis when melanocytes colonize the hair. Melanocytes within the morphogenetic hair bulge and bulb are thought to become molecularly and anatomically distinct around P4 Sox10 overexpression affects McSC establishment then we would expect that the LPP melanocytes in Tg(DctSox10)/Tg(DctSox10) animals will decrease with age after P4. First, as indicated by the postnatal coat color, we confirmed in Tg(DctSox10) animals at P2 that regions of the coat unaffected by congenital white spotting contained hairs that were similarly pigmented in the hair bulb and shaft in comparison to +/+ littermates show that while LPP melanocytes are detected in Tg(DctSox10)/Tg(DctSox10) mice, their numbers are moderately reduced. By P7/8, the number of LPP melanocytes in hairs of Tg(DctSox10)/Tg(DctSox10) mice decreases further to less than half of those observed in Tg(DctSox10)/+ and +/+ animals (Tg(DctSox10) homozygotes confirms that while there may be fewer overall melanocytes in these animals due to the embryonic effects of Sox10 overexpression, their ability to colonize the hair bulge at P2 is maintained. However, in Tg(DctSox10)/Tg(DctSox10) mice, the decrease in LPP melanocytes over time, and their absence in adults, suggests that high Sox10 levels during melanocyte colonization of the hair follicle disrupts the establishment of McSCs.The fact that termates . Counts animals . The preTg(DctSox10) affects other melanogenic proteins within McSCs at P2, we evaluated the LPP expression patterns of TRP1, a known target of SOX10, and KIT receptor, and a protein not known to be downstream of SOX10. We find a noticeable depletion of DCT+/TRP1+ LPP melanocytes in Tg(DctSox10)/Tg(DctSox10) in comparison to Tg(DctSox10)/+ or +/+ mice (Tg(DctSox10) dramatically changes the KIT expression profile; while the majority of LPP melanocytes are KIThigh in wild type, this switches to KITlow in Tg(DctSox10) mice (Tg(DctSox10)/Tg(DctSox10) mice. Together these experiments demonstrate that although increased Sox10 expression does not affect the ability of melanocytes to produce normally pigmented morphogenetic hairs, it does result in changes in the expression status of the LPP melanocytes that may affect their establishment as McSCs.To examine whether +/+ mice . We also10) mice . Thus alTg(DctSox10) homozygotes precludes our ability to phenotypically assess them at this age, however, the fact that Tg(DctSox10)/+ mice exhibit changes in LPP expression profiles at P2 suggests that a closer look at Tg(DctSox10)/+ skins is warranted. First as expected, immunolabeling validates the presence of SOX10 in both LPP and bulb melanocytes of anagen (7dpp) hairs in Tg(DctSox10)/+ and wild type adult mice (Tg(DctSox10) homozygotes, no change in the total number of melanocytes per LPP was detected within anagen (7dpp) hairs of Tg(DctSox10)/+ animals in comparison to wild type /+ hairs revealed the presence of pigmented, often dendritic, cells within the McSC compartment. Ectopic LPP pigmentation was also detected in Tg(DctSox10)/Tg(DctSox10) hairs that remained pigmented into the adult hair cycle, but was rarely present in wild type hairs (Tg(DctSox10)/+ adult mice also exhibit changes in the expression pattern of TRP1 and KIT at anagen /+ hairs contain considerably more pigmented DCT+/TRP1+ LPP melanocytes with an accompanying decrease in the number of DCT+-only LPP melanocytes (Tg(DctSox10) also affects the DCT/KIT expression profile in adult mice with Tg(DctSox10)/+ hairs showing an increase in KITlo/pigment\u2212 and KITlo/pigment+ LPP melanocytes at the expense of those that are KIThi/pigment\u2212 (Sox10 expression drives the inappropriate differentiation of LPP melanocytes into mature pigmented melanocytes. Together with the observation that Tg(DctSox10)/+ animals also exhibit early hair graying we conclude that the Sox10 levels must be tightly regulated to maintain the integrity of the McSC population.Closer inspection of pe hairs . LPP melt anagen . In wildanocytes . Tg. Based on our observation that MITF is retained in the majority of LPP melanocytes at adult anagen (Tg(DctSox10)/+ animals display normal MITF expression with approximately half exhibiting ectopic pigmentation /Tg(DctSox10) are combined with a hypomorphic Mitf mutant allele, vga9Mitfvga9/+Tg(DctSox10)/Tg(DctSox10); Mitf mice proceed to gray prematurely like their Tg(DctSox10) homozygote counterparts (compare Tg(DctSox10) are ameliorated by reducing endogenous Sox10 with heterozygosity for a Sox10 null allele heterozygotes exhibit demonstrable changes in expression of differentiation markers, we evaluated whether vga9Mitf also affects this aspect of the Sox10 overexpression phenotype. Upon visual inspection, an increased severity of hair graying in vga9/+Tg(DctSox10)/+; Mitf over Tg(DctSox10)/+ mice is particularly noticeable as these animals approach one year of age (vga9/+Tg(DctSox10)/+; Mitf mice in comparison to either single heterozygote or +/+ animals (vga9/+Tg(DctSox10)/+; Mitf mice also display a noticeable expansion in the amount of ectopic pigmentation within the LPP in comparison to Tg/+ animals (vga9/+Tg(DctSox10)/+; Mitf mice express TRP1 and are pigmented. This phenomena is less pronounced in Tg(DctSox10)/+ mice and is not observed in vga9/+Mitf or +/+ mice line, suggests that MITF participates in the negative regulation of Sox10 or Sox10-dependent processes within the McSC.Since LPP melanocytes of animals . Hairs f animals . By immu+/+ mice , 6E. TheSox10 is critical postnatally for the establishment and maintenance of cells in the melanocyte lineage. The role of Sox10 is twofold\u2014first, it is necessary for the retention of mature bulb melanocytes and undifferentiated McSCs, and second, it is required for the production of normal follicular pigmentation. The apparently contradictory requirements for Sox10 by undifferentiated McSCs and in the differentiation of melanocyte progenitors can be explained through our evidence that the McSC is maintained through the modulation of Sox10 levels itself. Accordingly, while SOX10 is expressed at all stages of the melanocyte lineage in mouse, increased Sox10 levels results in premature differentiation of the McSC population eliminating their capacity for self-renewal. This observation supports the idea that Sox10 activity within the McSC is normally decreased, and we provide evidence suggesting that this may occur through a MITF-mediated negative feedback loop. From these observations we propose a model for McSC establishment during early postnatal development whereby melanocytes migrating into the morphogenetic hair assume either a stem or differentiated cell fate depending on the environment they colonize. In this model, the hair follicle bulge would activate unique stem cell-specific signaling pathways in resident melanocytes, including one involving MITF-mediated negative feedback. Subsequent downregulation of Sox10 would establish the McSC. Melanocytes that colonize the hair bulb would not be subject to these repressive signals, Sox10 activity would remain high, and these melanocytes would undergo differentiation. This mechanism is also applicable to the maintenance of the McSC and production of pigmented melanocytes during adult hair cycling.In this study we show that Sox10 can contribute to the preservation of the undifferentiated McSC population while also driving melanogenesis is in agreement with current views on the ability of SOX proteins to confer different states of cellular maturity. Sox10, in particular, is credited for defining successive stages of neural crest cell development during embryogenesis; beginning with the maintenance of multipotency within the neural crest stem cell The idea that Sox10 levels are so precisely regulated within the postnatal melanocyte remains unclear. Previously, Sox10 expression was reported to decrease as melanocytes colonized the hair bulge leading to the speculation that establishment of the McSC is dependent on downregulation of Sox10Sox10 may provide survival of the postnatal melanocyte lineage, McSCs included, while a higher threshold of Sox10 expression is required to drive melanocyte progenitor differentiation and pigment production. This idea is similar to the Mitf rheostat model proposed by Carreira et al. Mitf expression can produce a range of melanoma phenotypes from stem cell-like to proliferative to terminally differentiated. While the precise mechanisms regarding Sox10 regulation are not fully known, conserved regulatory regions have been identified for Sox10 and encompass binding sites for transcriptional activators including SOX9B, NOTCH, \u03b2-catenin, LEF1, MED1(PBP), ATF2, and TFAP2 Sox10 expression\u2014\u03b2-catenin remains in the cytoplasm of McSCs during telogen, but shuttles to the nucleus during anagen where it is sufficient to drive the melanocyte differentiation program The mechanism by which Dct expression Hes1 and Hes5 are expressed by melanoblasts, and that Notch signaling, is critical in their survival Sox10 and McSC maintenance.The above observations do not discount the possibility that stem and progenitor fates in the melanocyte lineage may also be explained by a combinatorial mechanism where the availability of SOX10, regulatory regions of its targets, or partner transcription factors influence the cell state. For instance, SOX10 functions synergistically with a number of cofactors, namely PAX3, MITF, and CREB, during the activation of downstream genes Sox10 by MITF was shown using mathematical modeling to explain the dynamics of melanocyte differentiation within zebrafish vitMitf/vit mice exhibit similar ectopic pigmentation and hair graying defects as we observed with vga/+Tg(DctSox10)/+; Mitf mice vga9Mitf reduces congenital white spotting but exacerbates hair graying in with Tg(DctSox10) suggests a role for MITF within the McSC that is unique from its role within the melanoblast. In regards to the latter, we believe the white spotting phenotypes observed in Tg(DctSox10) mice may be explained by increased Mitf expression. MITF directly binds and upregulates genes required for melanin synthesis and melanosome biogenesis, including Tyr, Trp1, Dct, SILV, MLANA, and GPR143CDKN1A (p21) and CDKN2A (p16) Chx10 mutant mouse reveal that inappropriate maintenance of Mitf within the retinal progenitor cells leads to their reduced proliferation, transdifferentiation into pigmented cells, and consequent micropthalmia Tg(DctSox10)/Tg(DctSox10) mice have small eyes that are rescued by haploinsuffiency for Mitf . Together these observations indirectly support the idea that the hypopigmentation observed in this Tg(DctSox10) line may be attributed to increased levels of MITF within melanoblasts inhibiting their proliferation and/or causing their inappropriate temporal differentiation.A number of observations support the idea that MITF may repress McSC differentiation. First, the possibility of a negative feedback mechanism for the regulation of Tg(DctSox10) mice fits with the assertion that overexpression of Sox10 drives the premature differentiation of McSCs. The increase in the percentage of LPP melanocytes that are TRP1+/pigment+ in hairs of adult Tg(DctSox10) heterozygotes compared to wild type animals confirms this. However, we also observed an unexpected change in KIT receptor expression in LPP melanocytes with Sox10 overexpression. At adult anagen, the majority of bulge melanocytes in wild type mice exhibit high KIT immunofluorescence intensity (KIThi) and those in Tg(DctSox10) mice appear KITlow. Previous reports show that McSC progenitors rely on KIT signaling for their appropriate proliferation and pigmentation during hair growth, and bulge melanocytes that retain a KITlow/\u2212 status represent the McSC population. Sox10 produces numerous pigmented, Kitlow bulge melanocytes, suggests that regulation of melanocyte lineage differentiation can also occur independent of high KIT expression. This idea is supported by the observation that Kit mutants, when treated with ionizing radiation, produce ectopic pigmentation within the hair bulge and exhibit hair graying Kit, and this is exemplified in recent microarray studies showing that Sox10 knockdown in melanoma cells results in no significant change in Kit expression mice were rederived on and maintained by outcross to C57BL/6J tm1sorRosa26 mice were obtained as homozygotes, maintained by intercross and bred together with T2TYR::CreER mice to generate compound heterozygotes flSox10 and vga9Mitf mice were rederived on C57BL/6J and maintained by intercross Tg(DctSox10) line . vga9Mitf and LacZSox10 alleles were detected using PCR primers for \u03b2-galactosidase, 5\u2032-GATCCGCGCTGGCTACCGGC-3\u2032 and 5\u2032-GGATACTGACGAAACGCCTGCC-3\u2032, using the same PCR conditions described above. Primers and cycling conditions for the flSox10 allele was described previously Tg(DctSox10) transgene was determined by TaqMan analysis for two SNPs flanking the transgene on chromosome 1 that distinguish the original FVB donor strain from the C57BL/6 background strain (rs13475895 and rs13475987).Mice were genotyped using DNA isolated from tail tips and PCR analysis. Primers for the TAM was dissolved in corn oil or a combination of ethanol and sunflower oil. TAM treatment was performed by IP injection of lactating mothers or adults with 2 mg/animal for the number of days indicated.Morphogenetic and adult hairs were staged according to After shaving, skin from the lower back was immersed in 2% formaldehyde, and irradiated in a 540W variable wattage microwave three times in intervals of 30 s irradiation followed by 60 s on ice. After microwaving, samples remained in fixative for an additional 25 minutes on ice. Skins were cryoprotected in 10% sucrose overnight, embedded in NEG-50 (Thermo Scientific), frozen and sectioned with a cryostat (10 \u00b5m). For brightfield imaging, eosin-Y was sometimes used as a counterstain.Sections for immunolabeling were first rinsed in PBS with 0.1% Tween 20. For nuclear antigens sections were subjected to antigen retrieval by boiling for 20 minutes in a Tris-EDTA solution and then permeabilized by treating with 1% Triton X-100 for 15 minutes. Sections were blocked for two hours in 1% bovine serum albumin (Sigma) and incubated with primary antibody overnight at 4\u00b0C. Primary antibodies include those against DCT , SOX10 , PAX3 , MITF , c-KIT , TRP1 and TYR , Cre recombinase , \u03b2-galactosidase , and cleaved Caspase-3 . After washing, sections were incubated in the appropriate secondary antibodies for two hours at room temperature.Sequential immunolabeling was performed for co-detection of DCT and SOX10 as these antibodies were both generated in goat. After labeling for SOX10 using the protocol described above, sections were blocked with rabbit \u03b1-goat IgG FAB for two hours, washed and then labeled for DCT as described above.Brightfield and fluorescence microscopy was performed on a Zeiss Observer.D1 compound microscope. Images were obtained with an Axiocam Hrc camera using the Axiovision 4.8.2 software and processed with Adobe Photoshop. Quantitation of hair and cell phenotypes of immunolabeled tissue was performed on every fourth section of sequentially obtained skin sections. Data is presented as the mean \u00b1 standard deviation. Student's T-test with Bonferroni correction was used to determine statistical significance, unless stated otherwise.Skin samples were fixed in 2% formaldehyde/0.2% glutaraldehyde for 30 min at room temperature. Samples were then washed with rinse buffer (2 mM MgCl2/0.1% NP40/PBS) and stained overnight in X-Gal solution consisting of 0.32 mg/ml X-Gal, 5 mM ferrothiocyanide, and 5 mM ferrithiocyanide in rinse buffer.Tg(DctSox10)/+ mice was reverse transcribed using the High Capacity cDNA Reverse Transcription Kit (ABI). Quantitative PCR was performed using Taqman Fast Universal PCR Master Mix (ABI) and the following Taqman gene expression assays: Sox10 (Mm01300162_m1) and Pax3 (Mm00435493_m1). All experiments were performed with technical and biological replicates of \u22653.RNA from E17.5 skins from wild type and Figure S1+ melanocytes (red) present in the skin at P2. Skins from T2; Rosa26tm1sorTyr::CreER reporter mice treated at P2 and P3 with tamoxifen were harvested and analyzed for Bgal (blue) activity at P14 and 7 days post plucking (7dpp). Bgal+ cells are visible in the LPP (arrows) and bulb of the hair at both timepoints confirming that induction of T2Tyr::CreER perinatally successfully targets McSCs and their more differentiated progeny. (D) Double immunolabeling of these 7dpp skins reveals that 97% of perinatally lineage-marked Bgal+ cells (green) that exist with the LPP are DCT+ melanocytes .Perinatal melanocytes give rise to the McSC population. (A) Anti-Cre recombinase (green) is present in the majority of DCT(TIF)Click here for additional data file.Figure S2+ melanocytes located in the LPP and bulb of the hair follicle also express SOX10 . Arrows indicate examples of double-labeled cells.SOX10 expression in follicular melanocytes. Immunofluorescence staining of skins harvested at P2, P6, P14, 7dpp and 21 dpp reveals that the majority of DCT(TIF)Click here for additional data file.Figure S3MITF expression in follicular melanocytes. Immunofluorescence staining of skins harvested at P2, P6, P14, 7dpp, and 21dpp for DCT and MITF . Double-labeled cells are apparent in the LPP and bulb of the hair from P2 through 7dpp, but are not visible in melanocytes at 21dpp. Arrows indicate examples of double-labeled cells.(TIF)Click here for additional data file.Figure S4TRP1 expression in follicular melanocytes. Immunofluorescence staining of skins harvested at P2, P6, P14, 7dpp, and 21dpp for DCT and TRP1 . TRP1 expression is visible in hair bulb melanocytes throughout hair cycling, but is variable in LPP melanocytes. At P6 very few LPP melanocytes express TRP1, but this number increases through P14 and 7dpp and then remains relatively static during 21dpp. Arrows indicate examples of double-labeled cells.(TIF)Click here for additional data file.Figure S5+ melanocytes are detected at catagen, shown at 21dpp. Arrows indicate examples of double-labeled cells.TYR expression in follicular melanocytes. Immunofluorescence staining of skins harvested at P2, P6, P14, 7dpp, and 21dpp for DCT and TYR . In general TYR expression is detected most strongly in the melanocytes that exist in the hair bulb, and very rarely in LPP melanocytes. Few TYR(TIF)Click here for additional data file.Figure S6+, but with variable fluorescence signal intensity. At 7dpp, KIT highlights the dendricity of some LPP melanocytes. In the bulbs of P6, P14 and 7dpp hairs KIT expression is strongly localized to the keratinocytes at the bulb tip and KIT . In the LPP of hairs at P2, P6, P14 and 7dpp nearly all melanocytes are KIT(TIF)Click here for additional data file.Figure S7Sox10 loss results in a corresponding loss in MITF expression. (A\u2013B) Control animals do not exhibit a hypopigmentation phenotype; (A) fl/+; Tyr::CreERT2Sox10 animal treated with topical tamoxifen at P2\u20133 and imaged at P14, (B) untreated fl/fl; Tyr::CreERT2Sox10 animal imaged at P43. (C) Triple-labeling of hair bulbs from fl/fl; Tyr::CreERT2Sox10 mice described in +/MITF+/SOX10+ and PAX3+/MITF\u2212/SOX10\u2212 melanocytes, respectively. (D) Distribution of melanocytes double-labeled for PAX3 and MITF within pigmented (gray) and non-pigmented (white) hair bulbs in skins from fl/flSox10 and fl/fl; Tyr::CreERT2Sox10 harvested on 7dpp from mice treated with TAM on 0\u20133dpp (*p<0.0083).(TIF)Click here for additional data file.Figure S8Tg(DctSox10) results in an increase in Sox10 expression, and belly spotting, but no change in the MITF expression profile of LPP melanocytes. (A) Quantitative RT-PCR of e17.5 back skin confirms that Tg(DctSox10)/+ mice exhibit a 2.4-fold increase in Sox10 mRNA levels in comparison to wild type when normalized to Pax3 . Pax3 was used for normalization to control for possible variation in melanoblast numbers between genotypes. (B) The white belly spot observed in Tg(DctSox10)/+ mice varied in size from 0\u201310 white hairs up to a white spot measuring 7\u00d715 mm. The penetrance of the white belly spot in Tg(DctSox10)/+ mice was 97% (29/30). Occasionally, a few white belly hairs were also observed in the background strain, C57Bl/6 . (C) SOX10 (green) is evident within the LPP and bulb melanocytes of both +/+ and Tg(DctSox10)/+ mice. (D) Brightfield and corresponding fluorescent images of anagen III/IV hair follicles double labeled for DCT and MITF in wild type and Tg(DctSox10)/+ animals. (E) Comparison of the number LPP melanocytes per anagen III/IV hair follicle in +/+ and Tg(DctSox10)/+ animals that express DCT, and MITF, and produce ectopic pigmentation (*p<0.0001). The average number of MITF+ melanocytes per LPP was not significantly different between Tg/+ (2.81\u00b10.97) and wild type (2.78\u00b10.19) animals (p\u200a=\u200a0.96).(TIF)Click here for additional data file.Table S1+ melanocytes doubled labeled with the indicated marker per LPP+UTP or Bulge+SHG . Grayed cells indicate the combination with the highest percentage of cells doublelabeled. *SOX10 expression in melanocytes at 21dpp is present, but weak, and normal staining protocols for DCT/SOX10 double labeling diminished visible SOX10 signal. Thus melanocytes at this timepoint were identified using KIT and then double labeled for SOX10. LPP, lower permanent portion of the hair; UTP, upper transitory portion of the hair; dpp, days post plucking; SHG, secondary hair germ of the hair.Quantitation of melanocyte immunolabeling during hair morphogenesis and hair cycling. Percentage of DCT(PDF)Click here for additional data file."} +{"text": "No study has examined this pathway in the broader context of psychosis symptom severity. We investigated the association between symptom severity scores and expression of 25 Wnt pathway genes in blood from 19 psychotic patients. Significant correlations between negative symptom scores and deshivelled 2 (DVL2) and glycogen synthase kinase 3 beta (GSK3B) were observed. No gene expression levels were associated with positive symptoms. Our findings suggest that the Wnt signaling pathway may harbor biomarkers for severity of negative but not positive symptoms.Genes in the Wnt (wingless)/ Thus, we sought to correlate blood-based gene expression of the Wnt/\u03b2-catenin signaling pathway with negative and positive symptom severity indices of patients with a history of psychosis . We were guided by previous research that has examined the relationship between psychosis symptom severity and other gene expression pathways in peripheral blood [The Wnt (wingless)/elopment . Numerouisorders . While pal blood , 4 and hal blood .Clinical and gene expression data from 19 subjects meeting The Scales for the Assessment of Positive and Negative Symptoms (SAPS-SANS) were admhttp://www.genome.jp/kegg/). These genes included v-akt murine thymoma viral oncogene homolog (AKT1), adenomatous polyposis coli (APC), axin 1 (AXIN1), catenin beta 1 (CTNNB1), disrupted in schizophrenia 1 (DISC1), Dickkopf homologs 1, 3, and 4 , disheveled homologs 1\u20133 (DSL1\u20133), frizzled homologs 1\u20138 (FZD1\u20138), glycogen synthase kinase 3 alpha (GSK3A) and beta (GSK3B), low density lipoprotein receptor-related proteins 5 (LRP5) and 6 (LRP6), transcription factor 4 (TCF4), and wingless-type MMTV integration site family member 1 (WNT1). Although many genes that were not included may provide further insight into the Wnt signaling pathway as it relates to psychotic symptoms, we were careful to select both up- and downstream genes within the pathway in an effort to provide an adequate proxy of Wnt pathway functioning. The number of genes that populate the Wnt/beta-catenin signaling pathway is continually changing, and a consensus on the number has not been achieved. We focused on 25 of the most well-characterized genes involved in the Wnt signaling pathway, based on the Kyoto Encyclopedia of Genes and Genomes database of the 25 WNT signaling pathway genes examined were nominally correlated with scores on the SANS however, only DVL2 remained significant after Bonferroni correction. DVL2 showed a negative correlation whereas, GSK3B was positively correlated with the SANS with negative symptoms. Post hoc exploration of the four subscales of the SANS revealed significant negative correlations between DVL2 expression and affective flattening and alogia severity. GSK3B expression was positively correlated with alogia only. None of the 25 gene transcripts examined significantly correlated with severity scores on the SAPS. We were unable to perform qRTPCR validation for our genes of interest due to inadequate mRNA quantities although previous work with the current sample has successfully validated genes using qRTPCR [Two [http://www.stanleygenomics.org/) suggest that DVL2 is upregulated in postmortem brains of patients with greater than 5000\u2009mg of lifetime antipsychotic exposure. In addition, lithium [GSK3B in the mouse and rat brain [n = 10) of the 19 subjects in the current study were taking one or more of these medications at the time of assessment but post hoc adjustment for presence of one of these medications showed no difference in our reported results. Therefore, these results serve as preliminary evidence that the Wnt signaling pathway may harbor biomarkers for (primarily negative) symptom severity in psychotic disorders. Future longitudinal studies designed a priori to examine gene expression prior to, during, and after a psychotic episode and initiation of pharmacological treatment would clarify the biomarker potential of this pathway for psychosis symptom severity. Finally, it remains unclear whether expression in peripheral blood accurately reflects that found in the brain [DVL2 and GSK3B using more sensitive techniques is also warranted. If replicated, these results provide potential first steps toward development of objective tools for characterizing psychosis symptom profiles in clinical practice.Several lines of investigation have reported links between genes associated with the Wnt signaling pathway and schizophrenia , 15, mosme level , had a lt shown) for nega lithium as well at brain , 20. Morhe brain . HoweverOur results support previous work and provide preliminary data suggesting the Wnt signaling pathway as one of potentially many pathways harboring transcriptomic biomarkers for negative symptom severity in psychosis."} +{"text": "High performance liquid chromatography (HPLC) and electrophoresis are commonly used to diagnose various hemoglobinopathies. However, insufficient information about the transfusion history can lead to unexpected and confusing results. We are reporting a case of Juvenile myelomonocytic leukemia (JMML) in which HbHPLC was done to quantify fetal hemoglobin (HbF). The chromatogram showed elevated HbF along with a peak in the HbD window. A transfusion acquired peak was suspected based on the unexpectedly low percentage of HbD and was subsequently confirmed using parental HbHPLC. It is well known that incomplete history on test request forms sent to laboratories and inappropriate patient samples can lead to wrong diagnosis and hazardous consequences. Hemoglobin electrophoresis and High Performance Liquid Chromatography (HPLC) are routinely done to diagnose and classify hemoglobinopathies. Acquired and inherited conditions in which abnormal HPLC result can be seen include high fetal hemoglobin in Juvenile Myelomonocytic Leukemia (JMML), Diamond Blackfan Anemia (DBA) and Fanconi Anemia.9/L and platelet count of 16 \u00d7109/L. Peripheral smear showed 35% monocytes with an absolute monocyte count of 20 \u00d7109/L. Diagnosis of Juvenile myelomonocytic leukemia was suggested and Hb HPLC was advised to look for increased fetal haemoglobin. High Performance Liquid Chromatography (HPLC) was done using BioRad Variant II instrument with beta thalassemia short program. Hb HPLC chromatogram showed a raised HbF (11%) with HbA0 69.2%, HbA2 1.8% and a peak in D-window of 10.2% with a retention time of 4.06 minutes (A 7 year old female presented to the outpatient department with complaints of hepatosplenomegaly and lymphadenopathy for 3 months. Hemogram showed a hemoglobin of 9.5 gm/dl, total leukocyte count of 57.24 \u00d710 minutes . The exp minutes . The traThere are only a few reports of abnormal hemoglobin peaks following blood transfusions.4The incidental finding of such abnormal peaks may cause diagnostic and therapeutic dilemmas even to the most experienced, particularly so in patients with suspected hemoglobinopathies who have received multiple blood transfusions. To avoid this rare problem some authors have suggested screening of blood donors for hemoglobinopathies."} +{"text": "The genomic variability of HIV viruses has impeded the development of globally relevant HIV vaccines with ability to induce strong and broad cellular and humoral immune responses. Many strategies, such as codon/RNA optimization, addition of highly efficient leader sequences, use of consensus or mosaic antigens, and electroporation (EP) have all been applied to enhance the breadth and magnitude of immune responses induced by DNA vaccines.Highly optimized clade B and C consensus envelope DNA vaccines (pEY2E1-B and pEY3E1-C) were developed. Their immunogenicity was compared to the immune responses induced by an optimized mosaic gp160 envelope vaccine (pMosEnv) or optimized primary clade B and C envelope vaccines (pPK61-14 and p96ZM651gp140-CD5). The immunogenicity of these constructs was studied in different murine models. All vaccines were delivered using electroporation. Antibody responses were determined by ELISA and cellular responses by IFN-\u03b3 ELISPOT.Antibody analysis supported that the consensus immunogens induced stronger clade-specific antibody response than the primary immunogens, while the mosaic antigen was a poor inducer of antibody responses. Compared to the primary vaccine constructs, both consensus and mosaic constructs were more potent at driving diverse cellular immune responses. The strongest cross-reactive immune responses against various consensus peptides libraries were induced by pEY2E1-B and pEY3E-C. The consensus immunogens were up to three times more potent at driving subtype-specific responses that recognized the different cross-clade immunogens. pMosEnv exhibited robust cellular responses when a PTE peptide set was used.We conclude that the highly optimized consensus immunogens may fold into a relatively native conformation and exhibit improved focus on antibody and T cell conserved regions, while the mosaic immunogen provides important T cell responses but does not preserve the native structure of envelope to allow for similar antibody responses in their current forms. Further exploration of these immunogens will be interesting."} +{"text": "Toxoplasma gondii. Thus, DSB repair mechanisms could be excellent targets for chemotherapeutic interventions. Recent genetic and bioinformatics analyses confirm the presence of both homologous recombination (HR) as well as non homologous end joining (NHEJ) proteins in this lower eukaryote. In order to get mechanistic insights into the HR mediated DSB repair pathway in this parasite, we have characterized the key protein involved in homologous recombination, namely TgRad51, at the biochemical and genetic levels. We have purified recombinant TgRad51 protein to 99% homogeneity and have characterized it biochemically. The ATP hydrolysis activity of TgRad51 shows a higher MK and much lower catk compared to bacterial RecA or Rad51 from other related protozoan parasites. Taking yeast as a surrogate model system we have shown that TgRad51 is less efficient in gene conversion mechanism. Further, we have found that TgRad51 mediated gene integration is more prone towards random genetic loci rather than targeted locus. We hypothesize that compromised ATPase activity of TgRad51 is responsible for inefficient gene targeting and poor gene conversion efficiency in this protozoan parasite. With increase in homologous flanking regions almost three fold increments in targeted gene integration is observed, which is similar to the trend found with ScRad51. Our findings not only help us in understanding the reason behind inefficient gene targeting in T. gondii but also could be exploited to facilitate high throughput knockout as well as epitope tagging of Toxoplasma genes.Repairing double strand breaks (DSBs) is absolutely essential for the survival of obligate intracellular parasite A broken chromosome if unrepaired leads to loss of genetic information and cell death. It is thus very important for a cell to identify and repair the damaged region. There are two major pathways by which double stranded DNA breaks (DSB) can be repaired. In homologous recombination (HR) mediated repair it relies on searching of extensive homologous stretches of DNA, whereas non homologous end joining (NHEJ) requires little or no homology. These two pathways compete with each other for repairing a DSB. It is a complex phenomenon and how cell decides which pathway to choose is not clear yet. Also different organisms show distinct usage frequency of HR over NHEJ during DSB repair. If a DSB is occupied by Ku70/80 hetero dimer, NHEJ follows. However, as soon as a DSB is formed, if the ends are resected by 5\u2032-3\u2032 exonuclease to form a long 3\u2032 ss DNA, HR sets into action. This is because the long ssDNA overhang inhibits the loading of Ku70/80 and recruits the recombination proteins Toxoplasma gondii belongs to the eukaryotic phylum Apicomplexa, which infects about one third world population and under immune compromised condition it can cause serious illness. This protozoan parasite shares a number of structural similarities with disease causing parasites namely Plasmodium, Cryptosporidium etc. Thus it has become a model system to do functional genomics. However efficient gene targeting is a challenge in this parasite. It has been observed that in this parasite gene targeting efficiency is very low as they demonstrate high degree of non homologous end joining T. gondii is the only protozoan parasite that harbors non homologous end joining mediated DNA break repair mechanism. A divergent eukaryotic parasite Trypanosoma brucei possesses Ku70/80, however TbKu70/80 function in telomere maintenance T. brucei cell extract T. gondiiT. gondii and Plasmodium falciparum show strikingly opposite choice in DNA repair pathways. While Plasmodium falciparum depends solely on HR and apparently lacks NHEJ, T. gondii prefers NHEJ. Here we report the mechanistic insights for differential repair choices between these two closely related lower eukaryotes. PfRad51 has been identified as a DNA repair protein and has been speculated to play major role during mitotic recombination T. gondii. However, with increase in homologous flanking ends, we observe an increase in targeted gene integration similar to the trend observed with ScRad51.TgRAD51 and ScRAD51 were transformed into the \u0394rad51 (LS402) strain to create MVS26 and NRY2 strains respectively. The empty vector pTA \u0394rad51 strain to generate NRY1. NA14 and NA14\u0394rad51 strains were used TgRAD51 or ScRAD51 to NA14\u0394rad51 strain to generate SSY1 and SSY2 strains respectively. To knockout Ku80 gene from a series of isogenic strains KANMX gene flanked by up-and down-sequences of KU80 ORF was amplified from an already existing yeast ku80\u0394 strain. The primer pair OSB127 (AGT CTA TTA GCG GAA GTA CC) and OSB128 (GAA CGT CCT CTA CCC ACG) resulted in 200 bp and 220 bp flanking sequence of KU80 on each side of KANMX gene. This linear KANMX cassette of 2220 bp was used to knockout KU80 from the rad51\u0394 (LS402) strain. The yeast expression vector pTA was transformed into the \u0394rad51\u0394ku80 (SSY3) to generate SSY4 strain. The yeast expression vectors harboring ScRAD51 and TgRAD51 were transformed into the \u0394rad51\u0394ku80 (SSY3) strain to create SSY5 and SSY6 strains respectively.The yeast strains used in this work are tabulated in Table 1. The yeast expression vectors harboring Toxoplasma gondii Rad51 was amplified from the cDNA library (provided by Prof. Vern Carruther) of T. gondii RH strain using hot start Kapa Hifi DNA polymerase (Kapa Biosystems) as described by the manufacturer. The forward primer OMKB65 (GGATCCATGAGCGCCGTCTCTCTTCAG) and reverse primer OMKB82 (GGATCC TCAGTTGTCTTCGTAGTCGCC) are complementary to the 5\u2032 and 3\u2032 ends of the coding sequence of TgRad51 gene and the underlined oligo nucleotide represents the BamHI sites incorporated in both the primers. The PCR product of size 1065 bp was first cloned into TOPO2.1 TA vector (Invitrogen). The resultant plasmid was digested with BamHI to release the insert containing TgRad51 gene and was ligated with pET28a (Novagen) to construct N-terminal His6 tagged recombinant protein. The insert was sequenced and submitted to the Gene Bank (JQ771675). All primers used in this study were purchased from Sigma Aldrich.The open reading frame encoding TgRAD51 gene was subcloned into BamHI site of the 2 \u00b5 expression plasmid, pTA under the control of GPD promoter. In a similar way, ScRAD51 was PCR amplified from the genomic DNA isolated from W303a strain using the forward primer OMKB90 (GGATCCTGTCTCAAGTTCAAGAAC) and the reverse primer OMKB88 (CTGCAGCTACTCGTCTTCTTCTC), the underlined oligo nucleotides represented BamHI and PstI sites incorporated in the forward and reverse primer respectively. The PCR product of size 1203 bp was cloned into the yeast expression vector pTA under GPD promoter. All the cloning was confirmed by sequencing.The 6 tag was transformed into Escherichia coli host strain Rosetta (DE3). The whole transformation mixture was added to 10 ml LB media containing chloramphenicol and kanamycin and incubated at 37\u00b0C for overnight. Next morning 5% of the primary culture was added to 100 ml fresh LB media (containing chloramphenicol and kanamycin) and tested for the IPTG induced expression of the TgRAD51 protein. After OD600 reached 0.8, protein was induced by IPTG (Sigma) and grown on Luria broth for 4 hours at 37\u00b0C. About 1.0 gm cell pellet was suspended in 3 ml lysis buffer containing 1 mg/ml lysozyme and protease inhibitor (PMSF). It was then lysed on ice by sonication using (SONICS Vibram Cell\u2122) giving ten 40 second bursts at 200 W with intermittent cooling. After lysis the cell debris was spun down by centrifugation for 45 minutes at 10,000 g at 4\u00b0C. The supernatant was applied to 0.75 ml 50% Ni-NTA agarose (Qiagen) and the loading flow through was collected. The resin was next washed with 16 volume of wash buffer . Further the column was washed with 5 volume of wash buffer 2 . Finally the required protein was eluted first with buffer containing 250 mM imidazole (5 column volume) and then with 400 mM imidazole . The aliquots from the indicated steps in the elution profile were separated using 10% SDS-PAGE and visualized by Coomassie brilliant blue G-250 (Bio-Rad). The protein fractions eluted with 400 mM imidazole were pooled and dialysed against the dialysis buffer containing 20 mM Tris HCl (pH\u200a=\u200a8), 1 mM dithiothretol (DTT), 5% glycerol. The concentration of the purified recombinant TgRad51 protein was determined by UV absorbance at 280 nm by using the extinction coefficient of 21025 M\u22121 cm\u22121 as calculated from the amino acid sequence by using ExPaSy Protparam tool. The purified protein was further confirmed by MALDI-TOF analysis and MS sequencing.The expression vector pET28a:TgRad51 having N-terminal His2). The reaction mixture was incubated at 37\u00b0C for 35 minutes and at every 5 minutes time interval a measured reaction volume was added (at 22\u00b0C) to purine nucleoside phosphorylase (PNP) reaction mix (manufacturer description) for 30 minutes. The ATP hydrolysis results in the formation of inorganic phosphate which was then reacted with MESG (2-amino-6-mercapto 7-methyl purine riboside) generating 2-amino-6-mercapto 7-methyl purine having an absorbance at 360 nm. This reaction is catalyzed by PNP. At a typical ATP concentration, the amount of inorganic phosphate released was plotted against different time interval and from the slope of the straight line, rate of the reaction was calculated. For determining the kinetic parameters the reaction was done at various concentrations of ATP (20 \u00b5M - 600 \u00b5M). The rate of the reaction was plotted using Graph pad Prism software against each concentration of ATP to determine the value of MK and catk.The rate of ATP hydrolysis of TgRad51 was determined using Enz Chek Phosphate assay kit . A typical reaction mixture was composed of 2 \u00b5M TgRad51 along with 30 fold molar excess of \u03c6xssDNA (New England Biolab) (60 \u00b5M) in presence of ATP containing reaction buffer . In order to detect endogenous level of TgRad51 and ScRad51 from NRY2, MVS26, SSY1 and SSY2, the protein was isolated according to the protocol as described in URA3 genes are separated by KANMX6. The first ura3 gene is having a HO endonuclease recognition site in it. This strain expresses HO endonuclease when grown on galactose medium. Initially single colonies from each of the three strains NA14\u0394rad51, SSY1 and SSY2 were grown to logarithmic phase in the medium containing glycerol as a carbon source. About 1000 cells of each strain were then plated on two kinds of medium; one of them containing glucose and the other containing galactose as carbon sources. Colonies were counted on both the conditions after incubation for 3 days at 30\u00b0C. Galactose induced expression of HO creates a double stranded break at one of the URA locus and the repair efficiency was calculated by measuring the ratio of the number of colonies grown on the medium containing galactose, compared to that grown on medium containing glucose (no HO induction). Since such DSB repair could be achieved either by GC or SSA, in order to determine the efficiency of repair choice, we had replica plated the cells grown on galactose medium on the plates containing G418 sulphate. This assay was done thrice and the mean value was plotted using Graph Pad Prism5.In NA14 strain, there is a cassette integrated in chromosome V where two consecutive KANMX6 gene was excised from pFA6a-KANMX6 plasmid NotI restriction enzyme. This fragment was cloned at the NotI site of pSD158 plasmid ADE2 gene. The 5\u2032ADH4 and 3\u2032ADH4 encompass 477 and 1527 base pair respectively. The cassette comprised of KANMX6-ADH4-ADE2-ADH4 was released from the modified pSD158 plasmid by SalI digestion which was subsequently used for gene targeting assay. This cassette was transformed to \u0394rad51 (NRY1), NRY2 and MVS26, where efficient gene targeting at the ADH4 locus by homologous recombination resulted in expression of ADE2 gene but did not confer resistivity to G418 sulphate. The transformed colonies were subsequently selected on SC-ade-trp plates. The colonies grown in individual strains were replica plated on G418 sulphate containing SC-trp plate. Colonies sensitive to G418 sulphate represents targeted integration of ADE2 gene, while G418 sulphate resistant colonies correspond to random integration. Finally we calculated the % gene targeting efficiency by using the following formulae:A 1536 base pair fragment containing \u0394rad51 strain was subtracted. Each experiment was repeated three times. In another gene targeting assay, we monitored the gene targeting efficiency with increasing stretches of homologous flanking sequences . For that purpose we started with SLY3 strain where the SBA1 gene was already insertionally inactivated by a KANMX6 cassette KANMX6 cassette by three different sets of primers positioned at 200 bp/500 bp/1000 bp upstream and 200 bp/500 bp/1000 bp down stream of the KANMX6 insertion. The primer pair OSB5 (TGCTACCCGCCTTCGAGTG) and OSB6 (CACATACAGTTCCATTACTTGAC) resulted in 200 bp flanking sequence on each side of KANMX6 cassette. Similarly OMKB193 (CTCAGAAGAATTTCGTAAATCGG) and OMKB194 (GGAGATGGTACCGGTTAAGCG) produced 500 bp flanking sequences of SBA1 on either side of KANMX and OMKB191 (TCACACGTCCGTCATGTCTAC) and OMKB192 (GTCCTGCAGGAGACTTATTAGC) amplified 1 Kb flanking regions on either side of KANMX6 cassette. These three different PCR products were individually transformed to \u0394rad51 (NRY1), NRY2 and MVS26 strain to knock out sba1 by homologous recombination and selected on SC-trp containing G418 sulphate. The number of G418 sulphate resistant colonies was measured. We compared the efficiency of targeted integration in the rad51\u0394 and rad51\u0394ku80\u0394 strains. The gene targeting efficiency was monitored in these strains background with increasing length of homologous flanking sequences (500 bp or 1000 bp). In NKY8 strain, the CHL1 gene was already insertionally inactivated by a HIS cassette. We had amplified the HIS cassette by two sets of primers positioned at 500 bp/1000 bp upstream and 500 bp/1000 bp downstream of the HIS insertion. The primer pair OMKB 211 (CAG CTC TCT AGC CAA CAG CAG) and OMKB 212 (CTT GCG TAT TAT CTA TAG CGG C) resulted in 500 bp flanking sequence on each side of HIS3 marker. Similarly OMKB 213 (CAC TCG TTG ACT AGT TCA GAG G) and OMKB 214 (GAC GAA CTT CAT GTG ACG GCT G) produced 1 Kb flanking sequences on each side of the HIS3 marker. These two PCR products were individually transformed into NRY1, NRY2 and MVS26 strain to knock out CHL1 gene by homologous recombination and the transformants were selected on SC-trp-his plate. The numbers of colonies were measured. Similar experiments were done with SSY4, SSY5 and SSY6 strain to replace CHL1 gene with HIS3 marker by homologous recombination and the transformants were selected on Sc-Trp-His plates. The numbers of His+ colonies were counted. Integration at the targeted loci was determined by PCR. Each experiments were repeated at least three times and was plotted using Graph Pad Prism with error bars.In each case the value obtained for www.toxodb.org) have identified three ORFs corresponding to putative RAD51 orthologs from ME49 strain (TGME49_072900), VEG strain (TGVEG_020310) and GT1 (TGGT1_112080) strain. Based on these sequences we have designed PCR primers and amplified TgRad51 ORF from RH strain using cDNA library as template (a kind gift from Dr. Vern Carruthers). Sequence comparison of the amplified ORF indicated that it is an ortholog of Rad51. We found little sequence polymorphism of RAD51 gene among the four strains of T. gondii . The expression system Rosetta (DE3)/pET:TgRad51 was used for the purification of TgRad51 protein. The expression vector pET:TgRad51 was transformed into Rosetta (DE3) cells, which has the pRARE plasmid that supplies the tRNA for six codons that are rarely used in Mass spectroscopic analysis confirmed the presence of TgRad51 protein and yielded a molecular mass of 39,398 Da in agreement with the molecular mass of 38,796 Da predicted by ExPasy Protparam tool. MS-MS analysis yielded sequence of 5 peptides corresponding to the predicted amino acid sequence of TgRad51 . The wesT. gondii is due to a compromised Rad51 protein activity, we determined the ATP hydrolysis activity of purified TgRad51 protein. We used EnzChek Phosphate Assay kit (Molecular Probes) to determine the rate of ATP hydrolysis of TgRad51. In this assay the inorganic phosphate when combines with the substrate MESG (2-amino-6-mercapto-7-methylpurine riboside) in presence of the enzyme PNP (purine nucleoside phosphorylase) produces ribose 1-phosphate and 2-amino-6-mercapto-7-methylpurine. Formation of the product changes the absorbance of the substrate from 330 nm to 360 nm which can be measured spectroscopically. kcat value (0.034 min\u22121) and Km value for TgRad51 (81.53 \u00b5M) were calculated using Graph Pad Prism. Our results indicate that the ATP hydrolysis activity of TgRad51 protein is indeed very weak.In order to investigate whether the low gene targeting efficiency in URA3 gene in chromosome V with a KANMX6 selectable marker in between present in the same chromosome 3 kb apart, or it can be repaired by single strand annealing (SSA), which results in deletion of a stretch of intervening DNA sequence. We created two strains SSY1 and SSY2 by transforming the expression vector pTAScRAD51 and pTATgRAD51 respectively to NA14\u0394rad51 strain. In order to assay the efficiency of DNA repair activity of TgRAD51 we created a single DSB in the ura3 gene by HO induction and counted the number of cell survival on galactose containing plate and compared to the number of cells on glucose containing plate. Our result shows that the repair efficiency of TgRAD51 (83%) is comparable to that of the wild type (96%), while NA14\u0394rad51 being 63% (KANMX6 cassette. We found that the majority of the survivors (77%) of SSY2 strain (TgRad51) are due to repair by SSA, which does not require the presence of Rad51 protein, gene conversion efficiency being only 23% of the total. On the contrary SSY1 strain (ScRad51) exhibited almost equal preferences for GC and SSA pathways, i.e. approximately 42% of its survival by GC and 58% by SSA efficiency of TgRad51. Repair of a DSB by GC relies on Rad51 mediated homology search. To this end we have used yeast as a surrogate model system where GC could be assayed accurately and quantitatively. We used a yeast strain NA14 having two copies of between . One of eing 63% . In orde% by SSA . Hence t% by SSA shows thT. gondii targeted integration is extremely less efficient. We wanted to investigate whether such less efficient gene targeting is due to the weak ATP hydrolysis activity of TgRad51, since gene targeting at the correct locus demands search of two homologous needles in the genomic hay stack. In order to appreciate the contribution of TgRad51 protein alone, from other Toxoplasma proteins presumably involved in HR pathway, we have used a surrogate yeast model, where the yeast RAD51 gene is deleted and is replaced by TgRAD51 gene. Thus, in this system, the main recombination protein (Rad51) is from T. gondii and all other auxiliary proteins are from yeast origin. We have engineered this system in such a way that it would lead us to investigate the frequency of targeted vs. random integration choices in the cell harboring TgRAD51. The integration construct contains two selectable markers: ADE2 and KANMX6; while the ADE2 marker scores for all the integration events (targeted or random), the KANMX6 marker is able to differentiate between random versus targeted integration. In case of targeted integration at the ADH4 locus the KANMX6 marker will be lost as a result of homologous recombination. On the other hand, during NHEJ mediated random integration the KANMX6 cassette will be retained and such transformants will be G418 resistant. Thus random integration of the cassette would result in ADE2+ and G418R colonies ; NRY2 strain (\u0394rad51 containing an expression vector pTA: ScRAD51); and MVS26 strain (\u0394rad51 containing an expression vector pTA: TgRAD51). The experiments with three repeats showed that NRY2 cells (ScRAD51) prefer to go for targeted integration at the ADH4 locus with about 85% frequency, where as TgRAD51 showed a preference for random integration having only 24% frequency for targeted integration of SBA1 gene (SBA1 gene in the cells bearing ScRAD51 (NRY2) and TgRAD51 (MVS26). As expected, our result showed that with the increase in homologous sequence there was a gradual increase in the frequencies of the targeted gene disruption (\u0394rad51) cells showed very less number of targeted disruptions as measured by the survival on G418 sulphate plates throughout varied degree of homology. With increase in flanking homologous sequences, NRY2 cells (ScRAD51) showed a steep rise in the gene targeting efficiency (TgRAD51) cells behaved much like \u0394rad51 strain up to 500 base pair flanking homology. As the homologous stretches were increased to 1000 base pairs on either side of KANMX6, the number of G418R colonies also increased sharply for MVS26 strain. The increase in homologous flanking region causes same fold increase (approximately 2.5 fold) in the gene targeting efficiency for both ScRAD51 and TgRAD51.We transformed the KANMX-ADH4-ADE2-ADH4 cassette in three strains: NRY1 (\u0394egration . We hypoBA1 gene . These tsruption . NRY1 (\u0394ficiency . MVS26 (CHL1 locus in presence or in absence of Ku80. To this end we have tested six strains: MVS26 (TgRAD51); SSY6 (TgRAD51 \u0394ku80); NRY2 (ScRAD51); SSY5 (ScRAD51 \u0394ku80); NRY1 (\u0394rad51); and SSY4 (\u0394rad51 \u0394ku80). To disrupt CHL1 gene we have generated two constructs. These constructs contain a selectable marker (HIS3) flanked by upstream (500 bp/1000 bp) and downstream (500 bp/1000 bp) sequences of CHL1 ORF or single strand annealing (SSA). While SSA does not depend on Rad51, GC totally depends on Rad51 mediated homology search. Our finding that only GC is compromised in cells harboring TgRad51 is thus consistent. Two other assays (namely gene knock-in and gene knock-out) that also depend on Rad51 mediated homology search are also found to be affected by TgRad51.Several important findings of this study collectively help us in understanding the DSB repair pathways in E. coli, S. cerevisiae, S. pombe, Leishmania major and Plasmodium falciparum are 952, 465, 396, 297 and 231 respectively. Thus it is not surprising that all the aforementioned organisms prefer HR over NHEJ. The recombinase RecA (Rad51 ortholog) from most of these organisms have robust ATP hydrolysis activity catk of hRad51 is 0.16 min\u22121) T. gondii despite being a lower eukaryote shows preference to NHEJ pathway. The low gene density of Toxoplasma genome (100 genes per mega base) and our finding that TgRad51 too have a weak ATP hydrolysis activity (catk is 0.034 min\u22121) fits very well with the choice of DSB repair pathway in this organism. Although, it appears that HR might not be a general mechanism for repair of DSB in T. gondii, whether HR activity is required to repair specific types of DSB ends remains as an open question. It is likely that the primary function of HR in T. gondii could be during sexual reproduction.Double strand breaks (DSBs) can cause damage to the genomic integrity of a cell. Repair of such DSBs are essential for cell survival. A DSB can be repaired either by homologous recombination (HR) or by non-homologous recombination (NHEJ). Prokaryotes and lower eukaryotes prefer high-fidelity repair mechanism such as HR, where as higher eukaryotes show preference towards mutagenic NHEJ pathway. An attractive hypothesis links the repair choice to gene density. Due to the low gene density in higher mammals mutations acquired during NHEJ mediated repair of random DSBs, are less likely to affect gene function, where as in organisms with high gene density, the likelihood of having random DSBs within the genes are higher and concomitantly repair of such breaks by NHEJ might affect gene function. For example there are about 7 genes per mega base of mouse or human genome, thus the probability of having random breaks in the intergenic regions are higher than within the genes. On the other hand prokaryotes and lower eukaryotes have very high gene density. The gene densities (number of genes per mega base) of T. gondii to be characterized. The existence of putative orthologues of Dmc1, Rad54, Rad50 and Mre11 in Toxoplasma genome suggests that this parasite does possess a functional recombinosome. Interestingly, the apparent lack of Rad52 orthologue in T. gondii, Plasmodium falciparum and Cryptosporidium parvum is suggestive of a Rad52 independent recombination mechanism in these apicomplexan parasites.TgRad51 is the first member of the recombination machinery of T. gondii, targeted gene disruption or tagging of endogenous genes are very less efficient in this parasite. Works from Bzik laboratory and Carruthers laboratory have demonstrated enhanced gene targeting in ku80 null T. gondiiku80 null parasite lines via NHEJ mechanism or integration at the right locus via HR mechanism. Only one of the two types of recombinant clones (where integration took place at the right locus) is desirable. Abrogating NHEJ pathway (by knocking out KU80 or other molecular players of NHEJ) would definitely minimize the number of those recombinant clones where random integration has taken place and thereby facilitate the screening of the desired clone. However, it would not increase the integration efficiency at the correct locus. This notion is supported by the finding that targeted repair of \u0394hxgprt became independent of TgKu80 when enough flanking homology (910 bp) was provided T. gondii harboring Rad51 from L. major or S. cerevisiae could significantly increase the gene targeting efficiency in this protozoan parasite. Future studies might test this possibility either by expressing LmRad51/ScRad51 (having robust ATPase activity) in T. gondii from an episomal plasmid or by delivering purified Rad51 proteins via nano particle mediated delivery systems. Positive results from such studies will have a great impact on functional studies of Toxoplasma biology.Gene disruption and epitope tagging are the two most important tools to study gene function in the post genomics era. Both of these techniques rely on homologous recombination mechanism. Since the HR machinery is weak in Figure S1Phylogenetic analysis of eukaryotic Rad51 proteins using Clustal method . T. gondii Rad51 is highlighted.(TIF)Click here for additional data file.Figure S2Amino acid sequence of TgRad51 protein from RH strain. The underlined peptide sequences were generated from MS-MS analysis.(TIF)Click here for additional data file."} +{"text": "Dear Editor,The recent paper from Leonardi and La Rosa 2010) aimed to establish the prevalence of celiac disease (CD) in patients with chronic hepatitis B (HBV) infection 0 aimed t. Moreove"} +{"text": "Six patients with large or giant cerebral aneurysms were examined with 4D-Flow MRI and analyzed regarding 3D flow patterns and aneurysm wall shear stress (WSS). Two distinct groups of aneurysms were identified with fast swirling flow versus slow flow and significantly different WSS patterns, correlating with aneurysm morphology.Cerebral aneurysms are diverse and life threatening conditions. They typically develop at the major bifurcation sites of the intracranial vessels. In general, increasing size has been linked to higher rupture risk but specific data concerning which lesions will grow or de-stabilize is lacking. In patients with large and giant cerebral aneurysms, 4D-Flow MRI was employed to characterize hemodynamics and WSS patterns and their association with aneurysm location, shape and size.Six patients with large or giant cerebral aneurysms were studied. Aneurysms were located near the ICA bifurcation (n=4) with a saccular/spherical morphology or the basilar artery (n=2) with a fusiform morphology and examined using 4D flow MRI . Data were analyzed with in-house Matlab-based tools and 3D blood flow visualization software . Intra aneurysmal flow was visualized using time-resolved pathlines SCHN 1170/2-1."} +{"text": "We identify a Prdm14 and Nanog binding cis-acting regulatory region in Dnmt3b that is highly responsive to signalling. These insights provide a novel framework for understanding how signalling pathways regulate reprogramming to an epigenetic ground state of pluripotency.Genome-wide erasure of DNA methylation takes place in primordial germ cells (PGCs) and early embryos and is linked with pluripotency. Inhibition of Erk1/2 and Gsk3\u03b2 signalling in mouse embryonic stem cells (ESCs) by small molecule inhibitors has recently been shown to induce hypomethylation. We show by whole-genome bisulphite sequencing that 2i induces rapid and genome-wide demethylation on a scale and pattern similar to that in migratory PGCs and early embryos. Major satellites, intracisternal A particles (IAPs) and imprinted genes remain relatively resistant to erasure. Demethylation involves oxidation of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), impaired maintenance of 5mC and 5hmC and repression of the"} +{"text": "Intrinsic tooth discolorations after endodontic treatment are principally attributed to the composition of necrotic pulp tissue, hemorrhage within the pulp cavity, endodontic medicaments and/or filling materials. Residual sealer left in pulp chamber after obturation can cause discoloration. The objective of this in vitro study was to evaluate coronal discoloration created by AH26 and ZOE sealers after four months.Fifty intact human extracted maxillary central incisors were employed. Access cavities were prepared in all samples and root canals were instrumented; coronal orifices were then sealed using self-cure glass ionomer. The teeth were divided into two experimental groups (n=20) according to utilized sealer in pulp chambers including AH26 and Dorifill (ZOE). The remaining 10 teeth served as negative and positive controls (n=5). The access cavities were sealed with self-cure glass ionomer. Teeth were kept in incubator for four month. Preliminary digital images of the teeth were taken and then compared with those related to 4-month follow-up. The images were assessed using Photoshop software. Data was analyzed using paired t-test and independent samples t-test.The teeth which were filled with AH26 sealer showed significantly greater discoloration than those filled with ZOE sealer (Dorifill) (P<0.05).AH26 sealer causes greater discoloration of the crown compared to ZOE sealer. Despite the other disadvantage of AH26 sealer, it seems that Dorifill is more esthetically considerate. Anterior tooth discoloration is an esthetic problem for both the patient and dentist. Sources of coronal tooth discoloration can be natural (acquired) or iatrogenic (inflicted). Natural causes are those that occur as a result of tooth developing disturbances or due to patients\u2019 behavior, tooth caries, or traumatic injuries 2]3]4][3][4]2][[4][3][4][3][4]2]7].[7].6][7].[7].6]. Van. Van16].Results obtained from this study concurred with Van der Burgt et al. results; they showed that AH26 sealers cause a grayish discoloration and ZOE sealers cause a light red to orange discoloration . In thisIn this in vitro study, AH26 causes greater mean discoloration compared to Dorifill sealer after 4 months. Therefore, Dorifill (ZOE) sealers seem more appropriate for root canal treatment of anterior teeth. Similar studies with more samples and type of sealers and also periodical discoloration assessments are recommended."} +{"text": "Considering that the communication of the intestinal immunity with the systemic bloodstream can be a relevant adjuvant factor in the amplification of the host systemic inflammatory response and subsequent multiple organ dysfunction in sepsis, we aimed to evaluate the effect of the obstruction of the mesenteric lymph duct (OMLD) associated with massive fluid therapy in the early phase of sepsis and severe sepsis models.8 (S8) or 109 (S9) CFU/ml Escherichia coli inoculum intravenously (i.v.) (DL80 within 26 hours), and were treated with hyperhydration (HH) with or without previous OMLD (n = 5/group). Control group were na\u00efve animals (N) and animals submitted to HH or sepsis only. The mortality of groups was followed up to 30 days after experiments and microcirculation monitoring was observed at 6 hours post sepsis induction by videomicroscopy (sidestream darkfield imaging (SDF)).Adult Wistar-EPM rats were submitted to 10The effect of OMLD + HH reduced significantly the sepsis mortality rate: S8 60% to 14.5%) and S9 (80% to 60%). Besides, the liver and kidney microcirculatory features were better preserved as compared with untreated sepsis groups under video-microscopy (SDF) monitoring. (Figure 4.5% and These preliminary findings showed that both HH and OMLD have a potential therapeutic application in sepsis by minimizing the splanchnic organ's microcirculation dysfunction."} +{"text": "Parkinson\u2019s disease (PD) is the second most common neurodegenerative disease. Mutations in PINK1 and PARKIN result in familial forms of PD. We have found that lack of pink1 or parkin in Drosophila results in defects in mitochondrial integrity in multiple tissues. Flies lacking PINK1 or parkin show highly similar, if not identical, phenotypes.Genetic studies suggest that PINK1 and parkin function in a common genetic pathway with PINK1 positively regulating parkin. The PINK1/parkin pathway regulates mitochondrial dynamics by positively regulating drp1 and negatively regulating mitofusin. Mammalian cell-based studies suggest that the PINK1/parkin may regulate mitochondrial quality control via mitophagy. How autophagy intersects with mitochondrial dynamics in vivo will be discussed."} +{"text": "Hyaloperonospora arabidopsidis (Hpa) that are expressed during infection of its natural host Arabidopsis thaliana. Infection-related transcripts were identified from Expressed Sequence Tags (ESTs) derived from leaves of the susceptible Arabidopsis Ws eds1-1 mutant inoculated with the highly virulent Hpa isolate Waco9. Assembly of 6364 ESTs yielded 3729 unigenes, of which 2164 were Hpa-derived. From the translated Hpa unigenes, 198 predicted secreted proteins were identified. Of these, 75 were found to be Hpa-specific and six isolate Waco9-specific. Among 42 putative effectors identified there were three Elicitin-like proteins, 16 Cysteine-rich proteins and 18 host-translocated RXLR effectors. Sequencing of alleles in different Hpa isolates revealed that five RXLR genes show signatures of diversifying selection. Thus, EST analysis of Hpa-infected Arabidopsis is proving to be a powerful method for identifying pathogen effector candidates expressed during infection. Delivery of the Waco9-specific protein RXLR29 in planta revealed that this effector can suppress PAMP-triggered immunity and enhance disease susceptibility. We propose that differences in host colonization can be conditioned by isolate-specific effectors.Biotrophic plant pathogens secrete effector proteins that are important for infection of the host. The aim of this study was to identify effectors of the downy mildew pathogen R genes evolve to restrict further pathogen growth Plant pathogens secrete an arsenal of effector molecules that modulate host responses to enable successful infection. Effector proteins constitute part of the secretome of the invading organism and are regarded as being crucial for pathogenicity Some of the most highly co-evolved interactions are between plants and biotrophic pathogens. At one end of the biotroph spectrum are hemi-biotrophic species that initially colonize living cells but then switch to necrotrophy. At the other end are obligate biotrophs that maintain host cell integrity and depend entirely on their host for growth and completion of their life cycle. Obligate biotrophs have evolved sophisticated mechanisms for host cell manipulation and defense suppression Hyaloperonospora arabidopsidis (Hpa) naturally infects the model plant Arabidopsis thaliana, causing downy mildew disease Hpa is a highly specialized oomycete pathogen with a narrow host range. Analysis of the Arabidopsis-Hpa interaction has been particularly informative because of the extensive genetic variation in responses of different Arabidopsis accessions to a correspondingly diverse set of natural pathogen isolates Hpa isolates were found to be conferred by R genes often residing at polymorphic loci Phytophthora sojae (causing soybean root and stem rot), P. ramorum (sudden oak death) and P. infestans (potato late blight) Hpa isolate Emoy2 has also become available Hpa that are recognized by Arabidopsis R genes ATR1 and ATR13 polymorphism among Hpa isolates suggest that diversifying selection has driven the evolution of both genes The obligate biotroph Hpa pathogenicity genes by isolating pathogen transcripts expressed during infection have so far resulted in the isolation of only a few candidate effectors. A differential cDNA-amplified fragment length polymorphism (AFLP) analysis was used by van der Biezen et al. Hpa-infected Arabidopsis leaves leading to identification of 10 Hpa-derived cDNA fragments. In another study, a suppression subtractive hybridization (SSH) strategy was used to identify 25 Hpa-expressed genes from infected Arabidopsis Previous studies aimed at identifying Hpa-infected Arabidopsis leaves. To increase the proportion of Hpa transcripts in a mixed sample of plant and pathogen RNA we used a highly virulent Hpa isolate (Waco9) to infect a hyper-susceptible Arabidopsis mutant (eds1) Hpa unigenes we have identified and classified Hpa-secreted proteins into several effector categories based on the presence of characteristic domains/motifs. Eighteen of the Hpa proteins belong to the class of host-targeted RXLR effectors. While several effector candidates are shared with other Hpa isolates, and in some cases oomycete species, six predicted effector proteins from Waco9 are not detected in the genome of the sequenced isolate Emoy2, highlighting the dynamic nature of Hpa pathogenicity.Here we report a transcript sequencing approach based on Expressed Sequence Tag (EST) analysis of Hpa isolates Emoy2, Emwa1, Noco2 and Waco9 were assessed for growth on the Arabidopsis enhanced disease susceptibility 1 mutant Ws eds1-1Hpa isolate Waco9 compared to Emoy2 (to Emoy2 and otheto Emoy2 were theHpa and Arabidopsis (Hpa or Arabidopsis by comparing profiles of infected leaves with those of RNA from Hpa conidiospores mixed in different proportions with Arabidopsis leaf RNA (Hpa. Poly (A+) RNA was isolated from the infected leaf total RNA preparations and size-fractionated cDNAs ranging from 500 to 5000 bp were used for cDNA library construction.Total RNA isolated from the Waco9-infected material revealed ribosomal RNA (rRNA) peaks of bidopsis . The obsleaf RNA . This an7680 sequencing reactions corresponding to the 5\u2032 end of the cDNAs were performed. After vector trimming and removal of low quality sequences and chimeras, 6364 ESTs remained ranging in length from 50 to 1000 nucleotides (nt). The majority of ESTs had a read length of more than 500 nt . AssemblHpa, three oomycete pathogens and the NCBI nr nucleotide database. Almost all unigenes showed significant similarity (E<10\u22125) to known sequences. Whereas 1556 unigenes were identified as Arabidopsis sequences, 2164 unigenes had homologues within the sequenced Hpa isolate Emoy2 encoded proteins with a predicted signal peptide as analyzed with SignalP software \u22126) for transcripts encoding predicted secreted proteins when compared to the genome wide percentage of 6.8% and sequences in GenBank, excluding Hpa. Significant blast hits (E<10\u22125 and sequence coverage >75%) were obtained for 123 unigenes to search for sequences that might be missing from the Hpa genome assembly. No significant hits were obtained for six sequences, suggesting that these unigenes are present in Hpa isolate Waco9 but not in Emoy2.To define which of the unigenes . The remHpa unigenes predicted to encode secreted proteins was performed by comparison to the Pfam database of protein domains Hpa-specific .A functional annotation of the 198 specific . Homologspecific , indicaton motif , which hPhytophthora ELL-1 proteins Hpa. All three proteins have an extended C-terminal region following the elicitin domain, a feature of other oomycete ELLs Phytophthora ELIs/ELLs Elicitins (ELIs) are extracellular effectors characterized by a 98 amino acid conserved domain with a core of six cysteines in a specific spacing pattern allowing their classification into different groups. Elicitin-like proteins (ELLs) have more variation in the size and sequence of their elicitin domains Hpa secreted proteins, we determined the relative number of cysteine residues in the full-length translated sequences. A total of 16 HaCR proteins were identified which contain more than 5% cysteines and range in size from 72 to 405 amino acids (\u22125) was found to a protein of the ciliate Tetrahymena thermophila.A number of CR proteins secreted by fungal pathogens have been shown to be recognized by R proteins in resistant host plants no acids . Nine ofno acids . The cysHpa isolate Maks9 Notably, the HaCR sequences comprised the largest number of ESTs among the selected pathogenicity factors , indicatHpa unigenes encoding putative secreted proteins were therefore mined for candidate RXLR effectors. Proteins containing either an RXLR or RXLQ/RXLG motif in the mature protein were selected. Variations in the last arginine of the RXLR still allows protein translocation into the host cell RXLR proteins comprise a class of oomycete effectors that are translocated into the plant cell Hpa-specific . From the 14 Hpa-specific proteins, blast searches revealed that 10 RXLRs have corresponding genes in Emoy2 . RXLR5 was the least variable gene with identical proteins in all seven Hpa isolates.Coding sequences of the 18 RXLR candidate effectors were deduced from seven RXLR12 and 22 in Emoy2 or in any of the five other Hpa isolates, indicating that these are Waco9-specific. RXLR4 was only amplified from isolates Hind2 and Waco9. Although blast searches did not reveal a significant hit for RXLR16 in the Emoy2 genome DC3000\u0394CEL mutant strain which lacks the conserved effector locus (CEL) and is therefore unable to efficiently suppress PTI Pst DC3000\u0394CEL to induce callose deposition after infiltrating bacteria into leaves. In these experiments, YFP and ATR13 were expressed by the EDV vector in Pst DC3000\u0394CEL, respectively as negative and positive controls. Arabidopsis Col-0 exhibited numerous callose deposits after infiltration with Pst DC3000\u0394CEL-YFP system which was previously shown to successfully deliver the \u0394CEL-YFP A, B. ThPseudomonas bacteria. ATR13 enhances Pst DC3000 growth on Arabidopsis Pst DC3000 compared to delivery of YFP encoded predicted secreted proteins that are likely targeted to the host-pathogen interface and might therefore play a role in the infection process. Previous analysis of EST libraries generated from four developmental stages of P. sojae revealed the highest frequency of ESTs encoding secreted proteins in interaction libraries A total of 3729 unigenes were assembled consisting of 2164 Hpa secreted proteins, a putative function could be assigned to 90 sequences based on the presence of known domains/motifs, specific features (e.g. relative number of cysteine residues) or similarity to known proteins appear to be Hpa-specific sequences.It is notable that no putative function could be assigned for 108 unigenes corresponding to \u223c55% of the predicted secreted proteins identified in our study, and therefore these sequences were classified as unknowns . FurtherHpa proteins with a predicted function we identified three Elicitin-like proteins. Although ELIs and ELLs have been described in Phytophthora and PythiumSaprolegnia parasitica. The cysteine spacing pattern allowed classification of Phytophthora ELIs and ELLs in different groups Phytophthora ELLs including INL1 (P. infestans) and SOL1A (P. sojae) that belong to ELL-1 group. The spacing pattern of HaELL1 and 3 was not observed in Phytophthora. Further analysis of elicitins from different oomycete species should establish if there are species-specific patterns of cysteine spacing. Two beta-elicitins (cryptogein and cinnamomin) have been shown to bind lipids Phytophthora ELLs From the set of secreted Hpa-specific and no homology to other effectors specific ; Table 2Hpa-specific are not present in Emoy2, as determined by comparison with the Emoy2 genome sequence. Also, allele sequencing in seven Hpa isolates revealed that three RXLRs are so far only found in Waco9. The presence of an effector in some but not all isolates suggests that the gene has become lost during host-pathogen co-evolution to avoid triggering ETI. This has been described for effectors of different pathogens, including Avr4 from Phytophthora infestansCladosporium fulvumHpa effectors. Alternatively, isolate-specific effectors might affect colonization by different Hpa isolates by interfering mostly with host PAMP-triggered defenses. Our finding that RXLR29, an effector that is present only in the highly virulent Hpa isolate Waco9, is able to suppress PTI and enhance susceptibility to bacterial infection, supports this idea. The categorization of different effector protein types expressed by Hpa Waco9 during Arabidopsis leaf tissue colonization and before the developmental transition to asexual and sexual sporulation has allowed us to select effector candidates for further functional studies on interference with the host immune system. The activities and interactions of Waco9-derived effectors as well as the extensive allelic diversity of individual RXLR genes among Hpa isolates and oomycete pathogen species provides a rich source of material to trace the co-evolutionary history of this plant-biotroph system.We identified variations in the set of effectors produced by different Hpa isolates used in this study were originally collected from Arabidopsis populations within the UK and the Netherlands (Waco9) Hpa infections, conidiospore suspensions (5\u00d7104 conidiospores.ml\u22121) were spray inoculated on 2-week-old Arabidopsis seedlings. Plants were allowed to dry for 1 h and kept at 100% RH for 24 h in a growth chamber with 10 h light at 16\u00b0C. Plants were then moved to \u223c75% RH for an additional 4 to 6 days to delay asexual sporulation. Hpa growth on Arabidopsis leaves was visualized on whole-leaf mounts stained with trypan blue as described previously eds1-1 plants was isolated using the RNeasy Plant Mini Kit (Qiagen), following manufacturer's instructions. Poly (A+) RNA was purified from 1.3 mg of total RNA using dynabeads oligo (dT)25 . To eliminate rRNA contamination, the mRNA was purified twice. RNA concentrations were determined on a UV mini 1240 spectrophotometer (Shimadzu) and quality was assessed with the bioanalyzer 2100 using the RNA 6000 Nano Assay kit (Agilent Technologies). Poly (A+) RNA (5 \u00b5g) was used to construct a directional cDNA library in the \u03bb-Zap vector following manufacturer's instructions. cDNA was synthesized containing EcoRI and XhoI sites at the 5\u2032 and 3\u2032 ends, respectively, allowing unidirectional cloning of cDNA. Size fractionation of the synthesized cDNA's was performed and 12 fractions were collected and precipitated with 100% ethanol. The pellet was resuspended in RNase-free water and verified on the Bioanalyzer 2100 using the DNA 7500 LabChip Kit (Agilent Technologies). Phagemid DNA was excised without library amplification. DNA isolations and sequencing were done by Macrogen (Korea). Sequencing reactions were performed from the 5\u2032 end using a T3 promoter primer.For RNA isolations, infected leaf material was ground to a fine powder in liquid nitrogen. Total RNA of Waco9-infected Ws Hpa genome Phytophthora infestansP. ramorum and P. sojaeHa were submitted to dbEST (NCBI).Base calling, quality clipping and vector screening were performed with pregap4, which is part of the Staden sequence analysis package Hpa set of sequences was identified by translating the assembled EST sequences and singletons in 3 positive frames and selecting the longest ORF. Protein predictions shorter than 10 amino acids were discarded. For selection of the secreted proteins, all protein models were trimmed to start with a methionine. Signal peptide predictions were performed by SignalP version 3.0 http://www.cbs.dtu.dk/services/TMHMM/) Hpa genome was either taken from the Hpa genome paper http://www3.embl.de/cgi/pi-wrapper.pl. GPI anchor sites were predicted by the program big-PI Plant Predictor http://psort.hgc.jp/) HaELL, HaCR and HaRXLR genes were submitted to GenBank (accession numbers JF800099-JF800135).The most likely ORF of the Hpa or to isolate Waco9, similarity searches were conducted with BLASTP and TBLASTN with disabled low complexity filtering and a fixed database size of 1.000.000 rd June 2010) and a set of three oomycete genome sequences were selected and defined as Hpa-specific sequences. The predicted proteome and the genome assembly of Hpa isolate Emoy2 (versions 6.0 and 8.3) was searched to identify sequences potentially specific for isolate Waco9. All sequences with a significant hit and an identity ratio of >80% /length query) Hpa Emoy2 trace files that also contain reads not present in genome assembly. Sequences without significant blast hits were defined as Waco9-specific.For identification of secreted proteins specific to Hpa isolates Cala2, Emco5, Emoy2, Hind2, Maks9 and Noks1, genomic DNA was isolated from Hpa-infected Ws eds1-1 leaves using DNeasy Plant Mini Kit (Qiagen). Primer pairs and sequenced. Obtained sequences that were not readable due to amplification of different products of the same size, Phusion (Finnzymes)-amplified PCR products were cloned in pENTR/D-TOPO (Invitrogen) and sequenced. In cases where the obtained sequences were not full-length (3\u2032or 5\u2032 sequences missing), new primers were designed based on the Emoy2 genome sequence (Versions 6 and 8.3). When no PCR product was amplified, new reactions were performed using up to three new primer sets.To sequence the RXLR alleles of er pairs flankingThe RXLR DNA and predicted protein sequences were analyzed using Mega4 software Hpa Waco9 RXLR29 and YFP were amplified by PCR and cloned in pENTR/D-TOPO (Invitrogen) using primers: CACCATGGAGGTGGTCCTGATC (forward) and TTACTTGCCAGGACGCGC (reverse) for RXLR29 and CACCATGGTGAGCAAGGGCGAGGAGCTGTTC (forward) and AGTCTAGAGCTCTTACTTGTACAGCTCGTCCATGC (reverse) for YFP. Following Gateway cloning procedures these genes were cloned into pEDV6, a variant of the previously described pEDV3 E. coli DH5\u03b1 to Pst DC3000\u0394CEL or Pst DC3000-LUX, which has stable chromosomal integration of the luxCDABE operon from Photorhabdus luminescensE. coli HB101 (pRK2013) as a helper strain. Pst DC3000\u0394CEL-ATR13 (Emco5) and Pst DC3000-LUX-ATR13 (Emco5) were kindly provided by G. Fabro and J. Jones.The coding sequences of 8 cfu/ml Pst DC3000\u0394CEL suspensions. A total of \u223c48 leaf samples were taken for callose staining 12\u201314 h after infiltration. Leaves were cleared with 100% ethanol, re-hydrated and stained with aniline blue (0.05% in phosphate buffer pH8.0) for 24 h. Samples were analyzed with an Olympus AX70 Microscope using an UV filter. Callose spots were counted using the ImageJ software (http://rsb.info.nih.gov/ij/) Leaves of 5-week-old Arabidopsis accession Col-0 plants were hand-infiltrated with 1\u00d7105 cfu/ml. Initially, PstDC3000-LUX was used for assessment of bacterial growth by measuring increased luciferase activity as previously published Leaves of 5-week-old Arabidopsis accession Col-0 were hand-infiltrated with a bacterial inoculum of 5\u00d710Figure S1Growth of Hpa Emoy2 and Waco9 isolates in Arabidopsis. The level of colonization of cotyledons of 2-week-old Arabidopsis Ws eds1-1 seedlings infected with Hpa isolates Emoy2 and Waco9 at 7 dpi is visualized by trypan blue-staining and light microspcopy.(EPS)Click here for additional data file.Figure S2Determination of Hpa rRNA peaks on the Bioanalyzer 2100. Total RNA isolated from Hpa conidiospores and Arabidopsis leaves was mixed in different proportions to allow identification of the corresponding rRNA peaks of Hpa and Arabidopsis in bioanalyzer profiles. Peak sizes correlated with the relative amount of plant and pathogen total RNA.(EPS)Click here for additional data file.Figure S3Schematic representation of the Hpa Crinkler identified in the Waco9 cDNA library. The signal peptide (SP), the variable CRN motif (LYVAK) and the C-terminal domain (C) are shown.(EPS)Click here for additional data file.Figure S4Multiple alignment of RXLR29 sequences from 7 Hpa isolates. Insertion/deletion sites are indicated in yellow, stop codons are indicated in red.(EPS)Click here for additional data file.Table S1Primers used for RXLR allele sequencing.(XLS)Click here for additional data file.Table S2Enrichment of protein class members in the EST project compared to the genome-wide occurrence. A hypergeometric propability value <0.05 in the Enriched column indicates a significantly higher than average occurrence in the EST set of genes, e.g. all predicted secreted proteins .(XLS)Click here for additional data file.Table S3Functional classification of 90 unigenes based on Pfam domain predictions, BLASTX and manual annotation.(XLS)Click here for additional data file."} +{"text": "Vernicia fordii), whose seeds are rich in novel TAG with a wide range of industrial applications. The objectives of this study were to identify OLE genes, classify OLE proteins and analyze OLE gene expression in tung trees. We identified five tung tree OLE genes coding for small hydrophobic proteins. Genome-wide phylogenetic analysis and multiple sequence alignment demonstrated that the five tung OLE genes represented the five OLE subfamilies and all contained the \u201cproline knot\u201d motif (PX5SPX3P) shared among 65 OLE from 19 tree species, including the sequenced genomes of Prunus persica (peach), Populus trichocarpa (poplar), Ricinus communis (castor bean), Theobroma cacao (cacao) and Vitis vinifera (grapevine). Tung OLE1, OLE2 and OLE3 belong to the S type and OLE4 and OLE5 belong to the SM type of Arabidopsis OLE. TaqMan and SYBR Green qPCR methods were used to study the differential expression of OLE genes in tung tree tissues. Expression results demonstrated that 1) All five OLE genes were expressed in developing tung seeds, leaves and flowers; 2) OLE mRNA levels were much higher in seeds than leaves or flowers; 3) OLE1, OLE2 and OLE3 genes were expressed in tung seeds at much higher levels than OLE4 and OLE5 genes; 4) OLE mRNA levels rapidly increased during seed development; and 5) OLE gene expression was well-coordinated with tung oil accumulation in the seeds. These results suggest that tung OLE genes 1\u20133 probably play major roles in tung oil accumulation and/or oil body development. Therefore, they might be preferred targets for tung oil engineering in transgenic plants.Triacylglycerols (TAG) are the major molecules of energy storage in eukaryotes. TAG are packed in subcellular structures called oil bodies or lipid droplets. Oleosins (OLE) are the major proteins in plant oil bodies. Multiple isoforms of OLE are present in plants such as tung tree ( Vernicia fordii) is an economically important tree with a very limited growing area in the United States cis, 11trans, 13trans octadecatrienoic acid) Tung tree Triacylglycerols (TAG) such as tung oil accumulate in discrete subcellular structures called oil bodies in plants, similar to oil droplets in animals. Plant oil bodies mainly consist of TAG surrounded by a monolayer of phospholipids, with the hydrophobic acyl moieties of the phospholipids interacting with TAG and the hydrophilic head groups facing the cytosol OLE are a group of hydrophobic proteins localized on the surfaces of plant oil bodies founded primarily in the seeds and pollen. The precise functions of OLE are unknown. They may function to stabilize oil bodies at low water potential and/or regulate the sizes of oil bodies Prunus persica (peach) Populus trichocarpa (poplar) Ricinus communis (castor bean) Theobroma cacao (cacao) Vitis vinifera (grapevine) Arabidopsis OLE. Finally, we used TaqMan and SYBR Green qPCR assays to evaluate the relative abundance and tissue distribution of the five OLE mRNA in the seeds, leaves and flowers of tung trees.The objectives of this study were to identify OLE genes, classify OLE proteins and analyze OLE gene expression in tung trees. We identified five OLE genes in tung tree. We performed genome-wide phylogenetic analysis and multiple sequence alignment and classified the five tung OLE genes based on 65 OLE from 19 tree species including the sequenced genomes of 2 and stored at \u221280\u00b0C. The oil and fatty acid profiles of these tung seeds were reported previously by Cao et al, 2013 Tung trees were grown in the American Tung Oil Corporation orchard in Lumberton, Mississippi. John Corley, the Company officer, granted permission for this field study. Tung fruits were collected weekly for 11 weeks beginning June 23, 2006 (week 1). The developmental stage of week 1 seeds corresponded to approximately 9 weeks after full bloom and 1 month before the initiation of storage oil synthesis. Tung tree seeds were removed from the trees and kernels and immediately frozen in liquid NThe tung seed cDNA library used for EST analysis was constructed previously http://blast.ncbi.nlm.nih.gov/Blast.cgi) using tung tree OLE sequences. The properties and amino acid compositions of OLE were analyzed using Vector NTI software OLE sequences from other organisms were obtained from database searches using the keyword \u201coleosin\u201d and BlastP searches Total RNAs from tung seeds, leaves and flowers were isolated as described by Spectrum Plant Total RNA Kit (Sigma) 12\u201318 primer, 0.1 \u00b5g random primers, 500 \u00b5M dNTPs, 10 mM DTT, 40 u RNaseOUT, and 200 u SuperScript II reverse transcriptase in 1X first-strand synthesis buffer . The cDNA synthesis reaction was kept at 42\u00b0C for 50 min. The cDNAs were stored in \u221280\u00b0C freezer before qPCR analyses.The cDNAs were synthesized from total RNAs using SuperScript II reverse transcriptase as described Tms for the probes were approximately 10\u00b0C higher than the corresponding primers. They were synthesized by Biosearch Technologies, Inc . The amplicon sizes and the nucleotide sequences (5\u2032 to 3\u2032) of the forward primers, TaqMan probes (TET\u2013BHQ1) and reverse primers of OLE are described in PCR primers and TaqMan probes were designed using Primer Express software . The The optimized qPCR reaction mixtures contained variable amounts of total RNA-derived cDNA , 200 nM each of the forward primer, reverse primer, and TaqMan probe and 1\u00d7 Absolute QPCR Mix (TaqMan qPCR) or 1 x iQ SYBR Green Supermix (Bio-Rad Laboratories) without the TaqMan probes (SYBR Green qPCR) T method of relative quantification was used to determine the fold change in expression T) values of the target mRNAs to the CT values of the internal reference mRNA Rpl19b, Gapdh or Ubl in the same samples (\u0394CT\u200a=\u200aCTTarget\u2212CTref). The \u0394CT was further normalized with a calibrator, the sample control (\u0394\u0394CT). The fold change in expression was then obtained (2\u2212\u0394\u0394CT). The amplification efficiency of qPCR assay was estimated on the basis of the equation E\u200a=\u200a(10\u22121/slope\u22121)\u00d7100 The \u0394\u0394CAnonymous tung seed cDNA sequences from week 6 were generated and analyzed by pyrosequencing (\u201c454\u201d) technology as described previously Coffea arabica (coffee) Coffea canephora (coffee) Corylus avellana (hazelnut) Cocos nucifera Camellia oleifera (tea oil), 5 from Cupressus sempervirens (pencil pine) Citrus sinensis (orange) Elaeis guineensis Ficus pumila (climbing fig) Jatropha curcas (barbados nut) Juglans regia , 1 from Olea europaea (olive) Persea americana (avocado) Prunus armeniaca (apricot) Prunus dulcis Prunus persica (peach) Pinus taeda (loblolly pine) Populus trichocarpa (poplar) Ricinus communis (castor bean) Theobroma cacao (cacao) Vitis vinifera (grapevine) GenBank database search identified 75 unique OLE from 22 trees. These OLE include 1 from Arabidopsis contains 17 subfamilies of OLE including 5 forms of seed-specific OLE (S type), 3 forms of seed-and-microspore-specific OLE (SM type) and 9 forms of tapetum-specific OLE (T type) Arabidopsis genome Arabidopsis OLE and good amplification efficiency for all cDNA samples . SYBR Green qPCR, which utilizes different detection chemistry, was used to confirm the results from TaqMan qPCR assays. SYBR Green qPCR assays generated similar expression patterns in the three tissues technology using anonymous tung cDNA sequences from week 6 seeds. All five OLE shared significant conservation at the protein and nucleotide levels. The five OLE genes coded for small proteins with an average of 154 amino acid residues and an average of 16.5 kDa. They possessed high isoelectric point and high percentage of hydrophobic residues. These properties of OLE proteins were similar to those of the three DGAT in tung tree Prunus persica (peach) Populus trichocarpa (poplar) Ricinus communis (castor bean) Theobroma cacao (cacao) Vitis vinifera (grapevine) Arabidopsis OLE. Tung OLE1, OLE2 and OLE3 belong to the S type and OLE4 and OLE5 belong to the SM type of Arabidopsis OLE. The more abundant expression of OLE1, OLE2 and OLE3 than OLE4 and OLE5 in tung tree seed support the classification of tung OLE based on Arabidopsis OLE.The numbers of OLE genes in plants are widely different. We performed a genome-wide search for OLE genes in the sequenced tree genomes including Coffea canephoraBrassica napusIt is important to determine which isoform is expressed in developing seeds to understand the genetic control of oil biosynthesis and to guide rational design of successful transgenic plants. In this report, we analyzed expression of OLE gene family quantitatively in 3 tung trees, 3 tung tissues and 11 stages of seed development. Under optimized assay conditions, qPCR exhibited similar amplification efficiencies between the OLE genes and the ArabidopsisPhyscomitrella patensCoffea arabica and Caffea canephora; in addition, one OLE5 EST was found in the EST library of Caffea leaves C. reinhardtii grown under acetate-enriched medium It is noteworthy that OLE transcripts were detected in leaves and flowers of tung tree, although their mRNA levels were extremely low compared to those in the seeds. The expression of OLE genes in these non-seed tissues was verified by melt curve and gel electrophoresis analyses of the qPCR products with predicted sizes of the amplicons. It was generally accepted that OLE genes was only expressed in the seeds, pollen and tapetum but not in other tissues Arabidopsis OLE. Expression results demonstrated that all five OLE genes were expressed in developing tung seeds, leaves and flowers but their expression levels were much higher in the seeds than leaves or flowers. In addition, OLE1, OLE2 and OLE3 genes were expressed in tung seeds at much higher levels than OLE4 and OLE5 genes. Finally, the amounts of OLE mRNAs were rapidly increased in developing seeds and their expression levels were well-coordinated with tung oil accumulation. The information on OLE expression profiles suggests that OLE1, OLE2 and OLE3 genes may play major roles in tung oil biosynthesis and/or tung oil body development. Therefore, they might be preferred targets for tung oil engineering in transgenic plants.We identified five OLE genes in tung tree. Genome-wide phylogenetic analysis and multiple sequence alignment demonstrated that the five tung OLE represented the five OLE subfamilies and contained the \u201cproline knot\u201d motif (PX5SPX3P) shared among 65 OLE from 19 tree species. Tung OLE1, OLE2 and OLE3 belong to the S type and OLE4 and OLE5 belong to the SM type of There is no information about the genes coding for other oil-body proteins including caleosins and steroleosins in tung tree at this time. Identification and characterization of additional oil body proteins and their corresponding genes at the genomics and proteomics levels would enhance the understanding of genetic and mechanistic control of tung oil biosynthesis. This information is essential for improving important oils such as tung oil in the future.Figure S1Nucleotide sequence alignment of the five Ole genes in tung tree. Multiple sequence alignment was performed using the ClustalW algorithm of the AlignX program of the Vector NTI software. Ole sequence name is on the left of alignment followed by the GenBank accession number and the start of the nucleotide sequence of each Ole gene. The numbers at the top of the alignment are the positions of the multiple sequence alignment. The letters at the bottom of the alignment are the consensus nucleotides. Nucleotides in red on yellow represent those conserved in all five Ole sequences at a given position, whereas those in black on blue represent nucleotides conserved in majority of the sequences at a given position. The underlined nucleotides represent the forward primers, TaqMan probes and the complementary sequences of the reverse primers used in qPCR assays.(PDF)Click here for additional data file.Figure S2Multiple sequence alignment for the identification of amino acid residues and sequence motifs conserved in OLE. Multiple sequence alignment was performed using the ClustalW algorithm of the AlignX program of the Vector NTI software. Each OLE sequence name is on the left of the alignment followed by the position of amino acid residue of OLE protein sequence in the alignment. The letters at the bottom of the alignment are the consensus residues. Color codes for amino acid residues are as follows: 1) red on yellow: consensus residue derived from a completely conserved residue at a given position; 2) black on green: consensus residue derived from the occurrence of greater than 50% of a single residue at a given position; 3) blue on cyan: consensus residue derived from a block of similar residues at a given position; 4) green on white: residue weakly similar to consensus residue at a given position; 5) black on white: non-similar residues. The abbreviations of the organisms are: Car, Coffea arabica (coffee); Cca, Coffea canephora (coffee); Cav, Corylus avellana (hazelnut); Col, Camellia oleifera (tea oil); Citrus sinensis (orange); Egu, Elaeis guineensis ; Fpu, Ficus pumila (climbing fig); Jcu, Jatropha curcas (barbados nut); Jre, Juglans regia ; Oeu, Olea europaea (olive); Pam, Persea Americana (avocado); Pdu, Prunus dulcis ; Ppe, Prunus persica (peach); Pta, Pinus taeda (loblolly pine); Ptr, Populus trichocarpa (poplar); Rco, Ricinus communis (castor bean); Tca, Theobroma cacao (cacao); Vfo, Vernicia fordii (tung tree); Vvi, Vitis vinifera (grapevine).(PDF)Click here for additional data file.Figure S3Specificity of SYBR Green qPCR Assay. The qPCR reactions contained 5 ng RNA-equivalent cDNA from tung tree leaves and flowers. The qPCR products were separated by agarose gel electrophoresis. Lane 100 bp represents DNA ladders with 100 bp as the smallest band, increasing upward in 100 bp increments. The results using tung tree seeds are shown in (PDF)Click here for additional data file.Figure S4qPCR efficiency for OLE assay. TaqMan and SYBR Green qPCR reaction mixtures contained variable concentrations of RNA-equivalent cDNA from tung seeds, the optimized concentrations of each primer and probe (200 nM), and Absolute QPCR Mix (TaqMan qPCR) or each primer and 1 x iQ SYBR Green Supermix (SYBR Green qPCR). The results using RNA isolated from stage 4 seed of tree 1 are shown. The qPCR efficiency for Ole1 mRNA detection is presented in (PDF)Click here for additional data file.Figure S5Relative levels of OLE gene expression in developing tung seeds by TaqMan qPCR. The qPCR reaction mixtures contained 25 ng of RNA-equivalent cDNA from tung seeds and 200 nM of each primer and probe. The means of mRNA expression levels calculated from two qPCR assays in each seed stage using Rpl19b as the reference mRNA are presented in (PDF)Click here for additional data file.Figure S6Relative levels of OLE gene expression in tung tissues by SYBR Green qPCR. The qPCR reaction mixtures contained 5 ng of RNA-equivalent cDNA from various stages of tung seed, leaves and flowers and 200 nM of each primer. The means of mRNA expression levels calculated from two qPCR assays in each seed stage using Rpl19b as the reference mRNA are presented in (PDF)Click here for additional data file.Table S1TaqMan qPCR efficiency for quantifying Ole mRNA in tung tree tissues.(PDF)Click here for additional data file.Table S2Variation of Ole gene expression among tung trees.(PDF)Click here for additional data file.Table S3Ole gene expression among different stages of tung seeds.(PDF)Click here for additional data file."} +{"text": "Mice irradiated and reconstituted with hematopoietic cells lacking manganese superoxide dismutase (SOD2) show a persistent hemolytic anemia similar to human sideroblastic anemia (SA), including characteristic intra-mitochondrial iron deposition. SA is primarily an acquired, clonal marrow disorder occurring in individuals over 60 years of age with uncertain etiology.-/- and normal bone marrow cells using flow cytometry and gene expression profiling of erythroblasts. The predominant transcriptional differences observed include widespread down-regulation of mitochondrial metabolic pathways and mitochondrial biogenesis. Multiple nuclear encoded subunits of complexes I-IV of the electron transport chain, ATP synthase (complex V), TCA cycle and mitochondrial ribosomal proteins were coordinately down-regulated in Sod2-/- erythroblasts. Despite iron accumulation within mitochondria, we found increased expression of transferrin receptor, Tfrc, at both the transcript and protein level in SOD2 deficient cells, suggesting deregulation of iron delivery. Interestingly, there was decreased expression of ABCb7, the gene responsible for X-linked hereditary SA with ataxia, a component required for iron-sulfur cluster biogenesis.To define early events in the pathogenesis of this murine model of SA, we compared erythroid differentiation of Sod2These results indicate that in erythroblasts, mitochondrial oxidative stress reduces expression of multiple nuclear genes encoding components of the respiratory chain, TCA cycle and mitochondrial protein synthesis. An additional target of particular relevance for SA is iron:sulfur cluster biosynthesis. By decreasing transcription of components of cluster synthesis machinery, both iron utilization and regulation of iron uptake are impacted, contributing to the sideroblastic phenotype. Reconstitution of mice with hematopoietic stem cells deficient in manganese superoxide dismutase (SOD2) results in a hemolytic anemia that recapitulates many features seen in patients with sideroblastic anemia (SA) ALAS2) ABCb7) PUS1) was found to cause mitochondrial myopathy and SA GLRX5) as a cause of SA SLC25A38) have been identified in cases of hereditary SA SLC25A38 in a larger collection of hereditary SA samples confirmed that this gene is involved in a significant fraction (\u223c15%) of hereditary SA cases, although a large number of cases remain genetically undefined (>40%) The molecular genetic basis for several of the inherited forms of SA has been described. There are two X-linked sideroblastic anemias (XLSAs), one caused by mutations of an erythroid-specific form of the heme biosynthetic enzyme aminolevulinic acid-synthase have been found in patients with acquired idiopathic sideroblastic anemia (AISA) resulting in measurable respiratory chain dysfunction The majority of cases of SA are acquired, occur in the context of the bone marrow failure syndrome myelodysplasia (MDS) and remain idiopathic. In the small number of acquired SA cases where a defined genetic lesion has been identified, mitochondria are also implicated in pathogenesis. Somatic point mutations of mtDNA encoding subunit 1 of cytochrome c oxidase , a byproduct of mitochondrial respiration. SOD2-deficiency induces severe mitochondrial dysfunction in neurons and muscle leading to embryonic or neonatal lethality in homozygous mutant mice -/- HSC rescue irradiated wild-type recipients, however, Sod2-/- chimeric mice have a persistent hemolytic anemia. Sod2-/- red blood cells (RBC) show increased oxidative damage and have a shortened lifespan -/- chimeric animals) produced enhanced levels of reactive oxygen species (ROS) and showed increased protein oxidative damage, increased mitochondrial number and mass as well as mitochondrial hyperpolarization -/- erythroid progenitors.SOD2 is a mitochondrial antioxidant enzyme that detoxifies superoxide anion radicals were increased relative to normal (1.9-fold), and Sod2-/- CD71- TER-119Hi reticulocytes and mature RBC (R7) were decreased relative (5.9-fold) to normal compared to normal marrow, consistent with prior results We used a FACS-based method tracking expression of the erythroid-specific antigen TER-119 and CD71 (transferrin receptor) ntiation in murinper genotype) was labeled and hybridized on Affymetrix murine arrays to determine relative gene expression between normal and Sod2-/- erythroblasts.In order to better understand functional consequences of loss of SOD2 during erythropoiesis, we identified gene transcriptional changes in erythroid progenitor cells using cDNA microarray-based gene expression analysis. Erythroblasts were isolated from marrow of reconstituted mice using a sorting gate encompas-/- erythroblasts, while 327 were down-regulated. A scatterplot of comparative gene expression, with selected differentially expressed transcripts identified, is shown in -/- samples.The GeneSifter program was used to identify and categorize differentially expressed transcripts. Among the 476 differentially expressed transcripts 149 were expressed at higher levels in Sod2-/- erythroblasts comprised metabolic pathways with a direct role in mitochondrial metabolism or biogenesis. Nearly every enzyme in the tricarboxylic acid (TCA) cycle was down-regulated at the transcriptional level .We used gene ontology/pathway analysis al level . The excal level . Mitocho+/+ cells is decreased in Sod2-/- cells . No other genes associated with iron metabolism, heme biosynthesis or hereditary sideroblastic anemia showed significantly different expression between groups. and all transcripts showed the expected direction of change when comparing Sod2-/- and Sod2+/+ cells , GTP cyclohydrolase (Gch1\u20146 fold up) and tuberous sclerosis 2 (Tsc2\u20142 fold down).Array data were validated by performing TaqMan Gene Expression Assays on total RNA samples from independently prepared TER-119/+ cells . The val-/- cells In this study we have evaluated gene expression changes in erythroblasts that result from knockout of the antioxidant protein SOD2 in hematopoietic tissues\u2014a model previously shown to have many features of congenital or acquired sideroblastic anemia. Comparative FACS analysis using erythroid specific markers -/- erythroblasts. A prior study investigating erythroid development from hematopoietic stem cells deficient in the tumor suppressor Rb implicated PGC1\u03b2 as a regulator of erythroid development -/- cells either in microarray or subsequent qPCR experiments (data not shown). Expression of the related coactivator, PPRC1 -/- and normal erythroblasts on microarray implicating PPRC1 as a candidate regulator of mitochondrial biogenesis during erythroid development. While microarray analysis does not assess transcription of mitochondrial DNA-encoded subunits of the respiratory chain, we also found reduced expression of the critical nuclear encoded mitochondrial transcription factor A (Tfam) in Sod2-/- erythroblasts. Tfam is required for the expression of mitochondrial DNA-encoded genes, and is itself a target of PGC1\u03b1 -/- cells may also exert an effect on iron metabolism through a reduction in Fe:S cluster assembly\u2014and thereby impact iron homeostasis by altering the level of iron regulatory protein 1 (IRP1). Under conditions of iron deficiency, IRP1 activity increases as the apo protein (lacking an Fe:S cluster) accumulates. IRP's bind to regulatory sequences in the Tfrc mRNA 3\u2032 untranslated region and stabilize the messenger RNA\u2014promoting production of transferrin receptor protein ABCb7 has been reported in CD34+ cells isolated from MDS patients with idiopathic acquired SA (RARS), suggesting that ABCb7 is a critical, perhaps necessary target for the derangement of iron metabolism that leads to generation of ringed sideroblasts Ciao1-/- cells . The same argument can be made that reduced Ciao1 activity has potential to alter erythroid iron metabolism by tipping the balance toward generation of the apo protein, IRP1.Like human sideroblasts, SOD2 deficient erythroblasts do not effectively utilize iron creating a paradoxical situation in which cells are functionally iron-deficient despite iron overload. Microarray data revealed two differentially expressed transcripts involved in iron homeostasis, /- cells , and ABC-/- erythroblasts oxidative stress affects erythroid-specific transcriptional mechanism(s) that regulate expression of nuclear encoded mitochondrial proteins. One consequence of such dysfunction is altered iron metabolism secondary to impaired iron:sulfur cluster synthesis resulting from down-regulation of ABCb7 and Ciao1\u2014which impacts both mitochondrial iron utilization and regulation of iron metabolism. Identification of components of the mitochondrial biogenesis pathway active during erythroid differentiation will be an important step toward improving our understanding of the etiology of SA and will provide additional insights into the relationship between mitochondrial dysfunction and development of anemia. Novel insights into pathogenesis of sideroblastic anemia may come from evaluation of the function of additional candidate genes identified as strongly differentially expressed between normal and Sod2-/- erythroblasts, but with no previously defined role in erythroid development. Evaluation of these loci in both hereditary and acquired SA cases that lack mutations in previously identified SA genes may identify novel mutations associated with this disorder.At the level of gene expression profiling, the most prominent effect of loss of SOD2 is a broad down-regulation of multiple aspects of mitochondrial metabolism and biogenesis. This is consistent with current knowledge regarding causes of hereditary SA\u2014where mutations affecting proteins that localize to mitochondria, or mitochondrial DNA itself have been shown to be causal. SA is much more common later in life in the context of myelodysplasia, where etiology remains uncertain. Our results demonstrate that mitochondrial dysfunction (of diverse etiology) can be a primary event in development in SA. In Sod2All animal experiments were conducted under an approved protocol (ARC 06-0339) approved by the TSRI IACUC committee. All work complies with relevant federal and state regulations concerning use of animals.+/+ or Sod2-/- HSC were generated as previously described Female B6 mice reconstituted with Sod2+/+ and Sod2-/- samples were sorted in parallel using two FACSDiva and two FACSAria sorters (BD Biosciences). Ter119+, CD71+ erythroblasts corresponding to gate R4 in Bone marrow cell suspensions were labeled with a purified anti-CD16/32 mAb for cell surface Fc fragment receptors (FcR) saturation, FITC-conjugated anti-CD71 and PE-conjugated anti-TER-119 mAbs in ice-cold fluorescence-activated cell sorting (FACS) buffer (2 mM EDTA: 0.5% BSA: PBS). Cells were resuspended in sorting buffer (1 mM EDTA: 25 mM HEPES: 1% FCS: 10 \u00b5g/ml PI: PBS). Two pairs of Sod2+ cells were pulled down with streptavidin MicroBeads (Miltenyi Biotec) according to the manufacturer's instructions. Purity and cell morphology were evaluated by FACS analysis and microscopic examination of Wright-Giemsa-stained sorted cells cytospun onto slides, respectively. Ter-119+ cells represented 85.78%\u00b11.6% of magnetically-purified cells from whole marrow suspensions with 81.94%\u00b13.23% viability . We extracted and average of 8.29 (\u00b11.16) micrograms total RNA from 18.82 (\u00b13.90)\u00d7106 purified erythroid progenitors as material for validation using RT-qPCR reactions.RBC-depleted bone marrow cell suspensions were labeled with a biotinylated TER-119 mAb; TER-119).Cell suspensions were labeled with purified anti-CD16/32 (FcR saturation), FITC-conjugated anti-CD71 and PE-conjugated anti-TER-119 mAbs in ice-cold FACS buffer. 10 \u00b5g/ml PI was added prior to analysis on a FACSCalibur cytometer running CellQuest Pro (BD Biosciences). Gating distinct stages of erythroid development was baseTotal RNA was extracted using the RNeasy Mini kit accordingly to the manufacturer's suggested protocol for isolation of Total RNA from animal cells. Briefly, we disrupted and lysed 12.65\u00b11.90 million sorted cells (N\u200a=\u200a8) in RLT Buffer, homogenized and sheared genomic DNA through centrifugation on QIAshredder spin columns; we performed on-column DNA digestion with the RNase-Free DNase Set (Qiagen) and eluted total RNA in RNase-free water (Sigma-Aldrich).http://www.affymetrix.com/products/arrays/specific/mouse430_2.affx and http://www.functionalglycomics.org/static/consortium/resources/resourcecoree.shtml, respectively. GLYCOv2 oligonucleotide arrays are custom GeneChip arrays designed for the Consortium for Functional Glycomics .All experiments were performed using GeneChip Mouse Genome 430 2.0 and GLYCOv2 oligonucleotide arrays , as described at http://www.affymetrix.com/support/technical/manual/expression_manual.affx) as previously described in vitro transcription was performed with biotinylated UTP and CTP . Hybridization: the target cRNA generated from each sample was processed as per manufacturer's recommendation using a GeneChip Instrument System (Affymetrix). Briefly, spike controls were added to 6.7 \u00b5g and 10 \u00b5g fragmented cRNA before overnight hybridization on GLYCOv2 and 430 2.0 arrays, respectively. Post-processing and scanning: arrays were then washed and stained with streptavidin-phycoerythrin, before being scanned on a ScanArray 3000 (Affymetrix) using default settings and a target intensity of 250 for scaling. Quality control (QC): The amount and quality of starting total RNA and cRNA were checked with a ND-1000 Spectrophotometer and an Agilent Bioanalyzer , respectively, and/or on formaldehyde-containing agarose gels. After scanning, array images were assessed by eye to confirm scanner alignment and the absence of significant bubbles or scratches on the chip surface. GAPDH and \u03b2-actin 3\u2032/5\u2032 ratios were confirmed to be within acceptable limits although one Sod2-/- specimen, showed slight RNA degradation. Gene Sifter analyses performed with and without this 4th Sod2-/- sample yielded similar results, and all figures/tables are derived from a 4\u00d74 chip comparison. BioB spike controls were found to be present on all chips, with BioC, BioD and CreX also present in increasing intensity. When scaled to a target intensity of 250 using the GeneChip Operating Software (GCOS) v1.3.037 algorithm (Affymetrix), scaling factors for all arrays were within acceptable limits , as were background , Q values and mean intensities. Details of QC measures can be found at https://www.functionalglycomics.org/glycomics/publicdata/microarray.jsp, by clicking on the \u2018Raw Data\u2019 icon link for microarray experiment \u2018Jeff Friedman 1: Sod2 KO anemic mice\u2019.Total RNA from each sample was used to prepare biotinylated target RNA using standard Affymetrix protocol and were filtered upon the following criteria: a \u00b11.5 or 1.2 ratio cutoff , t-test with Benjamini and Hochberg MTC to adjust for false positives, FDR https://www.functionalglycomics.org/glycomics/publicdata/microarray.jsp; GLYCOv2 arrays), in NCBIs Gene Expression Omnibus and is accessible through GEO Series accession number GSE8726 (Mouse Genome 430 2.0 arrays). Gene ontology and KEGG pathway analyses were performed in GeneSifter to identify affected pathways. The entire gene expression dataset was used to query expression of sets of genes of interest .Data presented were generated using GeneSifter based pairwise analysis, RMA-normalized. CEL files and reverse transcribed using the High-Capacity cDNA reverse transcription (RT) kit . We performed quantitative real-time PCR (qPCR) using 30 ng cDNA template and the TaqMan Gene Expression Master Mix (ABI) in singleplex reactions on a 7900HT Fast Real-Time PCR System running SDS 2.3 (ABI) in default thermal cycling conditions and endpoint data collection. QC and amplification specificity of all targets was confirmed by melting curve analysis. Data analysis was performed using the RQ Manager 1.2 software (ABI) in \u0394\u0394Ct-tests were performed using GraphPad Prism ; results are shown as mean \u00b1 SEM; replicate numbers (N).Unpaired two-tailed Figure S1Heat Map of Mitochondrial Ribosomal Protein Gene Expression. 42 Distinct nuclear encoded mitochondrial ribosomal protein genes are expressed differentially when comparing Sod2+/+ and Sod2-/- samples. Data were filtered to display transcripts with \u22651.2 fold change and a corrected p value <0.05. Using these criteria, 43/45 significantly different transcripts are down regulated in Sod2-/- erythroblasts. 3 of the listed genes appear twice and are represented by 2 distinct probe sets.(DOC)Click here for additional data file.Figure S2Heat Maps Showing Genes Involved in: Iron Ion Homeostatis, Heme Biosynthesis, or Etiology of Sideroblastic Anemia. Expression of genes annotated as playing a role in iron homeostasis and heme biosynthesis is displayed for both Sod2+/+ and Sod2-/- samples without statistical filtering for fold change or significance. When statistical filters were applied to the set of iron homeostasis genes, ABCb7 and Tfrc were the only genes expressed with a fold change \u22651.5 and p value <0.05. ABCb7 is down in Sod2-/- cells, while Tfrc expression is up. Similarly, genes involved in heme biosynthesis are displayed without filtering on the right. When statistical filters were applied, a putative cDNA (Riken A230051G13) with a proposed role in glycine catabolism and heme biosynthesis was found to be expressed at higher levels in Sod2+/+ cells, meeting the same fold-change and statistical criteria. Finally, genes previously identified as mutated in hereditary sideroblastic anemia were queried for expression. Of these genes, only ABCb7 was (again) found to be significantly differentially expressed. Sod2 appears in the list of iron homeostasis related genes without showing differential expression. This reflects detection of expressed Click here for additional data file.Table S1-/- and Sod2+/+ Erythroblast Samples.List of 476 Differentially Expressed Transcripts Comparing Sod2 Criteria used in filtering data were fold change \u00b11.5 with a corrected p value <0.05 (Benjamini and Hochberg MTC used).(DOC)Click here for additional data file.Table S2KEGG Pathway Analysis of the 476 most highly differentially expressed genes. The GeneSifter program was used to generate a list of KEGG pathways using the 476 differentially expressed transcripts (fold change \u00b11.5 and corrected p <0.05) from table S2 that differ between groups. The top section of table shows results for all 476 differentially expressed transcripts, the middle panel shows analysis of only those transcripts that were expressed at higher levels in Sod2-/- cells, while the bottom panel shows only those transcripts that were expressed at lower levels in Sod2-/- cells. In evaluating the significance of identified pathways, a Z score greater than 2 is considered significant. However, the low number of identified genes in the set of transcripts with increased expression in Sod2-/- cells reduces confidence in some assignments. The strongest assignments are to metabolic pathways, splicing, and DNA repair. Several (strong) assignments are based upon overlapping gene sets (predominantly components of the oxidative phosphorylation pathway)\u2014for instance Parkinson's and Huntington's Diseases, where a mitochondrial link to pathogenesis has been identified.(DOC)Click here for additional data file.Table S3KEGG Pathway Analysis of Entire Microarray Dataset. As in table S3 above, GeneSifter was used to identify affected KEGG pathways utilizing the entire microarray dataset . This provides a much broader view of altered metabolic, signal transduction and disease processes that share patterns of gene expression change with those seen in our comparison of Sod2+/+ versus Sod2-/- erythroblasts.(DOC)Click here for additional data file.Table S4Taqman Assays Used for qPCR Validation:(DOC)Click here for additional data file."} +{"text": "Glioblastoma (GBM) is a highly malignant brain tumor with a dismal prognosis. Gene expression profiling of GBM has revealed clinically relevant tumor subtypes, and this provides exciting opportunities to better understand disease pathogenesis. Results from an increasing number of studies demonstrate a role for the immune response in cancer progression, yet it is unclear how the immune response differs across tumor subtypes and how it affects outcome. Utilizing gene expression data from The Cancer Genome Atlas Project and the Gene Expression Omnibus database, we demonstrate an enrichment of immune response-related gene expression in the mesenchymal subtype of adult GBM (n\u200a=\u200a173) and pediatric high-grade gliomas (n\u200a=\u200a53). In an independent cohort of pediatric astrocytomas (n\u200a=\u200a24) from UCSF, we stratified tumors into subtypes and confirmed these findings. Using novel immune cell-specific gene signatures we demonstrate selective enrichment of microglia/macrophage-related genes in adult and pediatric GBM tumors of the mesenchymal subtype. Furthermore, immunostaining of adult GBM tumors showed significantly higher cell numbers of microglia/macrophages in mesenchymal versus non-mesenchymal tumors (p\u200a=\u200a0.04). Interestingly, adult GBM tumors with the shortest survival had significant enrichment of microglia/macrophage-related genes but this was not true for pediatric GBMs. Consistent with an association with poor outcome, immune response-related genes were highly represented in an adult poor prognosis gene signature, with the expression of genes related to macrophage recruitment and activation being most strongly associated with survival (p<0.05) using CoxBoost multivariate modeling. Using a microglia/macrophage high gene signature derived from quantification of tumor-infiltrating cells in adult GBM, we identified enrichment of genes characteristic of CD4 T cells, granulocytes, and microglia/macrophages (n\u200a=\u200a573). These studies support a role for the immune response, particularly the microglia/macrophage response, in the biology of an important subset of GBM. Identification of this subset may be important for future therapeutic stratification. Glioblastoma (GBM), WHO grade IV astrocytoma, is a highly malignant disease with a poor prognosis despite aggressive treatment strategies Gene expression profiling of adult GBM has greatly aided our understanding of underlying tumor pathogenesis Although adult and pediatric high-grade astrocytomas are widely considered as having distinct clinical and molecular characteristics, a recent gene expression profiling study identified three major subtypes of pediatric high-grade glioma that recapitulated subtypes identified by Phillips et al. in adult high grade astrocytoma Several lines of evidence suggest that the immune response is important to glioma biology. First, there is a significant inverse correlation between atopic disease and serologic markers of allergy, including IgE, and the sporadic incidence of glioma The predominant immune cell infiltrate in human GBM consists of CD45+CD11b+ microglia/macrophages Given the clinical and experimental evidence supporting a role for inflammation in GBM, we combined transcriptional expression profiling and immunoprofiling data on human high-grade astrocytoma to investigate the tumor-associated immune response across tumor subtypes and in relation to disease outcome in adult and pediatric astrocytoma.Collection of samples used in this study was approved by the Human Research Protection Program Committee on Human Research of the University of California San Francisco. Each patient provided written consent for tissue collection, banking and research use by the UCSF Brain Tumor Research Center (BTRC).2 scale and quantile normalized between arrays. Duplicated probe signals were merged by taking their median values. The microarray data have been deposited in the GEO database (accession number GSE38330) and described in accordance with MIAME guidelines. Unsupervised hierarchical clustering was performed using Multiple Experiment Viewer Software . The gene signatures identified by Paugh et al. Total RNA was extracted from 24 flash-frozen pediatric astrocytoma samples described previously http://cancergenome.nih.gov) accessed on April 15, 2010. Gene expression data from the entire TCGA cohort of adult GBM patients (n\u200a=\u200a506) as of June 16, 2011 was downloaded from the TCGA Data Portal . Raw gene expression data from the entire TCGA cohort of adult GBM patients was downloaded from the TCGA Data Portal . The data was pre-processed by RMA procedure http://http://brainarray.mbni.med.umich.edu/Brainarray/Database/CustomCDF/CDF_download.asp, Version 14]. Raw gene expression data on 53 pediatric high-grade glioma patients (Affymetrix Human U133 Plus 2.0) http://www.ncbi.nlm.nih.gov/geo/, accession No. GSE19578) and was Log2 transformed and normalized using RMA.Gene expression data on 173 subtyped adult GBM patients http://www.broadinstitute.org/gsea), the molecular signatures database (http://www.broadinstitute.org/gsea/msigdb), and the C5: GO gene sets database, comprised of 1454 gene sets named by GO terms and contains genes annotated by that term (www.geneontology.org). For all GSEA analyses 1000 phenotype permutations were performed and a Signal2Noise ranking metric was used to create the ranked list of genes. A false discovery rate (FDR) q-value of less than 0.25 (25%) was considered statistically significant. There was minimal overlap of genes between the subtype-specific signatures and the immune response-related gene sets: 28 of 300 genes in the adult mesenchymal signature and 29 of 236 genes in the pediatric HC3/mesenchymal signature. Gene signatures suggestive of specific immune cell subsets (Table S1) were also used to analyze the data. Cell-specific gene signatures of M1 and M2 polarized macrophages were compiled from analyzing multiple studies characterizing M1 and M2 macrophages in vitro polarized human monocytes by Martinez et al. Gene set enrichment analysis (GSEA) http://www.rprojects.org). Microarray data used to develop the model included gene expression data of 58 genes derived from the GO terms Inflammatory Response (GO:0006954) and Response to Wounding (GO:0009611). Clinical variables included age at diagnosis, Karnofsky Performance Score (KPS) and gender. Overall survival was the outcome of interest and patients alive at the time of analysis were censored. The model was created using a training set of n\u200a=\u200a205 GBM patients from Carro et al. CoxBoost was designed to develop proportional hazards models from microarray data and clinical covariates using a boosting approach Formalin-fixed paraffin embedded tissue microarray\u2019s (TMAs) were generated from 34 adult GBM patients and 13 pediatric astrocytoma patients who had been transcriptionally profiled. Each tumor was represented at least twice on the array. Immunohistochemistry was performed according to standard methods and immunostaining for Iba1 , CA9 and CD34 was performed on the Ventana Medical Systems Benchmark XT. Iba1 cell counts were performed using ImageJ Software, where a minimum of three fields at 400\u00d7 magnification for each tumor were counted for Iba1 positive cells and then averaged. CA9 and CD34 staining was scored in a blinded fashion in a semi-quantitative manner as follows. CA9: 0, zero positivity; 1, <10% positivity; 2, 10\u201325% positivity; 3, >25% positive staining. CD34: 0, density of positive blood vessels less than normal brain; 1, density equal to normal brain; 2, density greater than normal brain. Mann-Whitney Rank Sum analysis, implemented in GraphPad Prism v5.0, was used to determine differences in immunostaining between patient groups. Correlations were performed using Spearman\u2019s rho. A test-for-trend via a linear regression model was used to evaluate a trend in Iba1 staining between subtypes in the direction of mesenchymal, classical, neural, proneural. A p-value of <0.05 was considered statistically significant.Iba1 scored tumors were divided into quartiles based on their microglia/macrophage cell counts and the highest quartile (high microglia/macrophages) was compared with the lowest quartile (low microglia/macrophages) for differential gene expression analysis. Significance Analysis of Microarrays SAM was usedTable S2A). Strikingly, 48% of the enriched gene sets in the mesenchymal subtype were related to immune response processes, and included gene sets associated with development of the immune system, lymphocyte activation, response to infection or injury, and the adaptive immune response. Gene sets associated with signal transduction ranked second (8%) in the mesenchymal subtype and, of these, 3 of 4 were related to NF-\u03baB signaling (Table S2A).Adult GBM of the mesenchymal subtype have upregulation of genes involved in interactions with the tumor microenvironment Table S2B). As the adult mesenchymal subtype proposed by Phillips et al. Figure S1A). Conversely, tumors defined as mesenchymal by Verhaak et al. Figure S1B). Thereby, demonstrating similarities between the adult Verhaak mesenchymal subtype and the pediatric Paugh HC3/mesenchymal subtype.Recent gene expression profiling of pediatric high-grade gliomas Table S3). In an independent cohort of pediatric astrocytomas from UCSF, we performed gene expression profiling , applied 1035 signature genes proposed by Paugh et al. Table S1) were identified from recent profiling studies Table S4A). The six enriched gene sets included gene signatures associated with microglia/macrophages, M1 and M2 macrophages and glioma infiltrating microglia/macrophages (GIM), along with the monocyte, granulocyte and na\u00efve CD4 T cell gene sets. Similar to adults, the HC3/mesenchymal subtype of pediatric GBM demonstrated significant enrichment of the gene signatures associated with microglia/macrophages and monocytes . Unlike adults, the gene sets representing granulocytes and na\u00efve CD4 T cells were not significantly enriched and two gene sets were negatively enriched; NK cells and nucleated erythrocytes . A similar analysis was performed with the independent UCSF cohort of pediatric astrocytomas. Due to the small number of GBMs (n\u200a=\u200a2) in the HC3/mesenchymal subtype , we included all astrocytoma grades in the analysis. Similar to the larger pediatric GBM data set, there was significant enrichment of the M2 macrophage gene signature and significant negative enrichment of nucleated erythrocytes , in the HC3/mesenchymal subtype (Table S4C).Having demonstrated an upregulation of immune response-related genes in the mesenchymal subtype we aimed to characterize the cell types potentially driving this pattern of gene expression. Immune cell-specific gene signatures . Microglia/macrophage recruitment can be regulated via multiple mechanisms including levels of HIF1\u03b1 Figure S3B). Subtype specific comparisons are shown in Figure S4.Increased expression of microglia/macrophage-related genes could reflect increased numbers of immune cells or increased transcript expression levels per cell. To determine whether the mesenchymal subtype contained a greater number of microglia/macrophages, we generated tissue microarrays from adult subtyped GBM (n\u200a=\u200a24) and from UCSF pediatric subtyped astrocytomas (n\u200a=\u200a13), and analyzed microglia/macrophage number, as determined by Iba1 immunohistochemistry. While there was inter-tumoral heterogeneity, the mean microglia/macrophage number was significantly greater in the mesenchymal subtype of adult GBMs than in the non-mesenchymal subtype . To identify the immune response-related genes in this list of 58 that are most closely associated with survival in a robust and multivariate manner, we used CoxBoost modeling Previous studies have identified a gene signature associated with worse prognosis in adult GBM Figure S5, eight of these immune response genes were expressed between 1.25\u20131.59 fold higher in the microglia/macrophage enriched population compared to bulk tumor. This fold change was comparable to the microglia/macrophage specific markers CD45, CD11b and Iba1 (1.28\u20131.69 fold).To assess if these survival-related immune response genes may be expressed by the microglia/macrophage population within the tumor mass we examined gene expression profiling data from tumor-associated cells enriched for microglia/macrophages compared with bulk tumor from a single GBM sample Table S6A). In contrast to adult GBM, a similar stratification of pediatric GBM into short and long survival groups did not reveal a significant enrichment of immune cell-specific gene signatures in the short survival group (Table S6B).Rather than stratify tumors by subtype, adult GBM patients (n\u200a=\u200a506) were stratified based on survival , and GSEA analysis demonstrated an enrichment of all macrophage, monocyte, and granulocyte gene signatures in short survival patients . Comparing tumors with the highest and the lowest number of microglia/macrophages . Eight of nine high microglia/macrophage tumors were in Cluster 3 and six of nine low microglia/macrophage tumors were in Cluster 2.To identify factors associated with increased microglia/macrophage infiltrates, we stratified adult tumors (n\u200a=\u200a34) by number of tumor-associated microglia/macrophages (Iba1) and analyzed them by immunohistochemistry and differential gene expression analysis. Ignoring tumor subtype, there was no significant association between microglia/macrophage number and levels of hypoxia or vascularity in our cohort of adult GBM , validating our immune cell type specific gene signatures. Furthermore, Cluster 3 had significant enrichment of the granulocyte and CD4 na\u00efve T cell signatures , suggesting a potential association between high microglia/macrophage infiltrate and increased expression of characteristic granulocyte and CD4 T cell genes.GSEA analysis of Cluster 3 versus Cluster 2 demonstrated significant enrichment of the M2 macrophage and monocyte gene signatures . Consistent with heterogeneity within tumor subtypes, Cluster 3 also contained non-mesenchymal tumors, including classical, neural and proneural tumors Click here for additional data file.Figure S2Distribution of tumor grade between tumor subtypes in two pediatric astrocytoma cohorts.(PDF)Click here for additional data file.Figure S3GSEA analysis of cell-specific gene signatures and number of microglia/macrophages in mesenchymal versus non-mesenchymal subtypes in the UCSF pediatric astrocytoma cohort.(PDF)Click here for additional data file.Figure S4Subtype specific comparisons of microglia/macrophage cell number, hypoxia and vascularity in adult and pediatric astrocytomas.(PDF)Click here for additional data file.Figure S5Expression of survival-associated immune response-related genes in glioma infiltrating microglia/macrophages relative to bulk tumor.(PDF)Click here for additional data file.Figure S6Correlation between microglia/macrophage cell number and hypoxia or vascularity in adult GBM.(PDF)Click here for additional data file.Table S1Cell-specific gene signatures used for GSEA analysis.(XLSX)Click here for additional data file.Table S2Enrichment scores and statistics of the top 50 enriched GO gene sets from GSEA analysis of the mesenchymal versus non-mesenchymal adult and pediatric astrocytoma subtypes.(XLSX)Click here for additional data file.Table S3Enrichment scores and statistics of defense/immune response GO gene sets from GSEA analysis of the mesenchymal versus non-mesenchymal subtypes in the UCSF pediatric astrocytoma cohort.(XLSX)Click here for additional data file.Table S4Enrichment scores and statistics of cell-specific gene signatures from GSEA analysis of the mesenchymal versus non-mesenchymal subtypes in adult and pediatric astrocytomas.(XLSX)Click here for additional data file.Table S5Gene Ontology analysis of the TCGA Worst Prognosis Signature.(XLSX)Click here for additional data file.Table S6Enrichment scores and statistics of cell-specific gene signatures from GSEA analysis of short versus long survival patients in adult and pediatric astrocytomas.(XLSX)Click here for additional data file.Table S7Enrichment scores and statistics of cell-specific gene signatures from GSEA analysis of Cluster 3 (surrogate high microglia/macrophage) versus Cluster 2 (surrogate low microglia/macrophage) in adult GBM.(XLSX)Click here for additional data file."} +{"text": "Nematodes and fungi are both ubiquitous in marine environments, yet few studies have investigated relationships between these two groups. Microbial species share many well-documented interactions with both free-living and parasitic nematode species, and limited data from previous studies have suggested ecological associations between fungi and nematodes in benthic marine habitats. This study aimed to further document the taxonomy and distribution of fungal taxa often co-amplified from nematode specimens. A total of 15 fungal 18S rRNA phylotypes were isolated from nematode specimens representing both deep-sea and shallow water habitats; all fungal isolates displayed high pairwise sequence identities with published data in Genbank (99\u2013100%) and unpublished high-throughput 454 environmental datasets (>95%). BLAST matches indicate marine fungal sequences amplified in this study broadly represent taxa within the phyla Ascomycota and Basidiomycota, and several phylotypes showed robust groupings with known taxa in phylogenetic topologies. In addition, some fungal phylotypes appeared to be present in disparate geographic habitats, suggesting cosmopolitan distributions or closely related species complexes in at least some marine fungi. The present study was only able to isolate fungal DNA from a restricted set of nematode taxa; further work is needed to fully investigate the taxonomic scope and function of nematode-fungal interactions. In benthic marine environments, free-living nematodes often outnumber other groups in terms of diversity and abundance and are known to play key roles in ecosystem processes Free-living nematodes are also known to harbor microbes on their body surface, dominated mostly by symbiotic bacterial phylotypes In a recent study we provided preliminary evidence of possible ecological association between fungi and free-living marine nematodes in the coastal sedimentary environments from southwest England based on 18S rRNA PCR-DGGE approach Suberites zeteki.No amplification of common airborne fungal spores was detected during the course of the study, confirmed by PCR control reactions. In total, 30 nematode specimens from Rame Head (RH) were identified based on morphological characters and subsequently amplified using previously reported nematode 18S rRNA primers (SSU_F04 and SSU_R22). Out of 30 nematodes, 12 co-amplified fungal 18S rRNA amplicons in addition to nematode 18S rRNA sequences. Three different fungal clones were detected alongside Rame Head nematodes: RH1 (three sequences), RH2 (five sequences) and RH3 (four sequences). The fungal sequences from Rame Head consisted of a 355 to 400 bp fragment from the extreme 5\u2032 end of the 18S rRNA. According to BLAST matches (Blastn), fungal sequences showed 98\u2013100% identity with other published fungal 18S sequences in GenBank and >97% identity to unpublished high-throughput environmental sequence data obtained from marine sediments . The seqIn total, 40 deep-sea nematode specimens were morphologically identified down to genus level and subsequently amplified using all four sets of nematode 18S primers. Fungal 18S rRNA sequences were detected alongside 18S rRNA amplicons from fifteen deep-sea nematodes in this study. Eight fungi-like phylotypes were detected alongside 18S rRNA amplicons from 11 nematode specimens when targeted with SSU_F04-SSU_R22, denoted as phylotypes DS1, DS2, DS3, DS4, DS5, DS6, DS7 and DS8. The fungal sequences varied between 347\u2013404 bp in length and showed 99\u2013100% identities with published fungal 18S sequences in GenBank and >95% identity to an unpublished high-throughput environmental dataset . In addiPhylogenetic approaches based on Maximum Likelihood (ML) method and NeigVerticillium sequences in a distinct clade that consisted of some other Verticillium-like species as well as with an uncultured fungal sequence with a marine origin. These were linked to, but distinct from, a group formed largely of species that were pathogens of other fungi. In both the phylogenetic methods the bootstrap support was very strong for this clade. Both RH1 and RH2 belong to the phylum Ascomycota based on the phylogeny.The fungal clone type RH1 clustered in a distinct group within a larger Chaetothyriales clade (94% and 95% bootstrap support in ML and NJ methods); some taxa within this group were also represented by marine genera. The fungal clone type RH2 was recovered with the Basidioradulum radula of the order Hymenochaetales and an uncultured Agaromycotina clone belonging to the phylum Basidiomycota in ML and NJ trees. Deep-sea fungal sequence type DS3 clustered with cultured fungal genera belonging to the order Xylariales under the phylum Ascomycota with significant bootstrap support. The deep-sea fungal sequence types DS4 and DS5 were placed into two separate clades containing cultured and multiple uncultured fungi of deep-sea origin of the order Tremellales and Ustilaginales respectively under the phylum Basidiomycota, bootstrap support was significant for DS4 and DS5 in ML tree. The remaining fungal sequence type DS6 formed a cluster with cultured members and uncultured fungal sequences in the order Botryosphaeriales under the phylum Ascomycota.Clone type RH3, along with deep-sea fungal 18S rRNA sequence types DS1 and DS7, were recovered in a distinct clade containing several uncultured fungal 18S rRNA sequences of diverse marine origin including amplicons from the gut of an amphipod from Pacific Ocean and anoxic sedimentary environment in the Arabian Sea to amplify a wide range of eukaryotic phyla from environmental samples For deep-sea nematodes, fungal sequences were most often obtained after specimens had been stored in slide mounts for long periods of time. Subsequent methodological investigations revealed that the slide mounting process can effectively degrade nematode DNA during storage Methods for extracting nematode genomic DNA are quite \u2018dirty\u2019 compared to protocols for larger organisms, where an animal's size may allow for DNA to be extracted from a subsample of clean tissue (e.g. muscle or whole appendages). Lysis of single nematodes involves the digestion of entire specimens\u2014thus nematode protocols inherently extract genomic DNA from all other organisms present in the nematode digestive tract and on the cuticle. Given the low amount of genomic material usually obtained from single nematodes, it is likely that fungal DNA and nematode DNA can act as competitive templates for primer binding during PCR reactions.Exophiala castellanii (syn. Rhinocladiella mansonii) have been isolated from nematode eggs Frequent co-amplification of fungus and nematode sequences implies a close relationship between these two taxa; however, the precise association between fungal species and marine nematodes is presently unclear. Fungal sequences obtained from individual nematode specimens may represent gut contents, detritus attached to the cuticle, or alternatively, fungal species that exhibit symbiotic or pathogenic associations with nematodes. Fungal-feeding behaviours are well documented in soil nematodes Fucus serratus thallii using the DGGE approach, with most isolates showing similarity to Lulworthia, Lindra, Sigmoidea/Corollospora, Acremonium/Emericellopsis and other ribotypes Fungal associations have been recorded in several eukaryotic phyla, and thus evidence for relationships with nematodes is not entirely unexpected. Several studies have reported the presence of endophytic mycophycobionts associated with brown seaweeds Terschellingia sp., Terschellingia longicaudata, Parodontophora sp., Viscosia viscosa, Sabatieria pulchra, Daptonema sp. and D. normandicum suggests that the fungal sequence types may be restricted to certain group of nematodes; our results also correspond with nematode taxa that were observed to have co-amplified fungal sequences in a previous study Both RH1 and RH2 fungal sequence types show striking similarity with fungal sequences detected previously from the coastal waters of southwest England using a PCR-DGGE approach Southerniella sp. showed 100% identity with an uncultured fungus clone FAS_91 18S (Acc No GU072590) sequenced previously from the oxygen depleted sediments of the Arabian Sea The 18S rRNA is relatively conserved in fungi and has been used to reconstruct deep evolutionary relationships in cross-phyla phylogenetic studies While we cannot confirm that highly similar fungal phylotypes represent the same biological species (given the conserved nature of 18S rRNA), our results suggest some intriguing patterns amongst marine fungi. Phylotypes exhibiting 100% pairwise identity to published sequences from remote locations may indeed represent broad geographic distributions for some fungal species; further work (including population genetic studies) will be required to confirm this hypothesis. In addition to geographic distance, these results suggest that some fungal species may adapt to different biogeochemical conditions . In an evolutionary context, highly similar sequences recovered from geographically disparate locations suggest (at minimum) closely related fungal species complexes that are globally distributed across marine environments.Morphological evidence has long suggested cosmopolitan distributions for some marine fungi The results from this investigation provide further evidence to support the existence of nematode-fungal associations in marine ecosystems. The recovery of identical or highly similar sequences in BLAST searches, coupled with geographic insight from 454 data, suggests that some fungal phylotypes are widespread and maintain cosmopolitan distributions. Despite these insights, a more intensive focus is needed to validate and quantify these putatively cosmopolitan fungal species. We currently do not understand the scope of nematode-fungal interactions, but limited data suggests a taxonomic bias for nematodes maintaining fungal associations. The ubiquity of both nematodes and fungi in benthic marine habitats suggests that both these taxa (and their interspecific relationships) fill key roles within these ecosystems. Finally, the present study highlights the utility of large environmental datasets for addressing specific biological hypotheses; high-throughput data generated in an independent study ultimately provided key insight towards elucidating the global distribution of marine fungal taxa. As sequencing costs continue to fall and more large datasets are generated, such datasets will likely play key roles in our understanding of global biodiversity.Fungal sequences were obtained during two independent investigations of nematode fauna: one study focusing on shallow water specimens collected off the UK Coast, and a second study investigating deep-sea taxa in the Southern Indian Ocean. Shallow-water sediments were collected off Rame Head (RH) (50m depth) , an offshore site close to Plymouth in South West England in 2004 and were immediately fixed in molecular grade ethanol for further studies. Deep-sea sediments were collected aboard RRS Discovery cruise D300 in December 2005, with the sample location representing an abyssal plain site in the Southern Indian Ocean (near the Crozet Islands). Deep-sea samples were obtained using a megacorer . Upon collection, the top 0-1 centimetre sediment fraction of each sample was immediately fixed in DESS preservative containing 5% DMSO. Nematodes analysed in this study were collected from site M6 of the CROZEX investigation; this particular sample area receives minimal organic flux from surface waters Nematodes were extracted from sediments via decantation and flotation in Ludox, following standard methods DNA was extracted following established protocol Nematode 18S rRNA primers from previous studies were used in this study 5\u2032-GCTTGTCTCAAAGATTAAGCC-3\u2032) and SSU_R22 (5\u2032-GCCTGCTGCCTTCCTTGGA-3\u2032)(i) SSU_F04 (5\u2032-TCCAAGGAAGGCAGCAGGC-3\u2032) and SSU_R24 (5\u2032-CCCCRRTCCAAGAATTTCACCTC-3\u2032)(ii) SSU_F22 (5\u2032-TCCAAGGAAGGCAGCAGGC-3\u2032) and SSU_R26 (5\u2032-CATTCTTGGCAAATGCTTTCG-3\u2032)(iii) SSU_F22 (5\u2032-AGAGGTGAAATTCTTGGATC-3\u2032) and SSU_R13 (5\u2032-GGGCATCACAGACCTGTTA-3\u2032)(iv) SSU_F24 . Plasmid DNA containing the inserts were cycle sequenced using BigDye Terminator Kit v3.1 . Cycle sequencing reactions were cleaned using the Wizard Magnesil\u2122 system . PCR products from deep-sea samples were cleaned using a QIAquick PCR purification kit (Qiagen), and cycle sequenced using a BigDye Terminator kit v3.1. Products from cycle-sequence reactions were cleaned via ethanol precipitation before being submitted for direct sequencing. For all specimens, sequencing was carried out in both directions using standard primers in an ABI Hitachi 3100 Genetic Analyzer. Generated sequences were then compared with known 18S rRNA sequences from GenBank and EMBL using the BLAST search engine (BLASTn program).http://www.arb-silva.de). The alignment was trimmed to the 5\u2032end part of the fungal fragments and was subsequently used for phylogenetic analysis. Phylogenetic tree based on maximum-likelihood (ML) with GTR+G+I model and gamma distribution was constructed in MEGA v5.0 Rhizomucor miehei (Acc No AF192506) (member of Fungi incertae sedis) was used as outgroup in both the trees. Environmental sequences reported in this paper have been submitted to GenBank and their accession numbers are from JF708047-JF708073.Sequences generated in this study were inspected closely to ensure that there were no PCR chimera events using CHIMERA-CHECK program http://datadryad.org/).Environmental pyrosequencing datasets Click here for additional data file.Table S1Geographic distribution of sequence reads contained within each top-scoring environmental OCTU, including number of raw sequences within each cluster.(DOC)Click here for additional data file.Table S2Location information for sites analysed as part of an independent 454 study of marine habitats.(DOCX)Click here for additional data file.File S1FASTA file containing OCTU consensus sequences of the top-scoring BLAST hits in an environmental 454 dataset.(TXT)Click here for additional data file."} +{"text": "Current standard therapy commonly followed for chronic Hepatitis C Virus (HCV) in Pakistan is interferon alpha plus ribavirin combination therapy (IFN \u03b1/ribavirin) and pegylated interferon plus ribavirin (PegIFN/ribavirin). PegIFN/ribavirin has increased rate of sustained virological response than standard IFN \u03b1/ribavirin therapy. Objective of current study was to analyze rate of early and delayed response to antiviral treatment as well as rate of relapse response in patients following standard treatment IFN \u03b1/ribavirin and in patients following pegylated interferon treatment.Baseline serum samples of 153 patients enrolled for IFN \u03b1/ribavirin and 50 patients for PegIFN/ribavirin were collected. After total RNA extraction, genotyping was and HCV RNA viral load was done. Subsequently HCV RNA viral load was estimated at 4 weeks of treatment, at 12 weeks, at 24 or 48 weeks and finally after 6 months follow up period. All the data was statistically analyzed using fisher's exact test.Total 86 patients out of 153 patients following conventional IFN \u03b1/ribavirin therapy completed treatment and 69% of them showed Rapid Virological Response (RVR). Whereas 50 patients following PegIFN/ribavirin treatment completed treatment and 80% of them achieved RVR. Total 64 out of 86 patients following IFN \u03b1/ribavirin therapy completed follow up period and 53.5% of them achieved Sustainded Virologcal Response (SVR). Forty-five out of total 50 patients who received PegIFN/ribavirin treatment completed 6 months follow up period and among these 70% achieved SVR. SVR rates were significantly associated with RVR (p < 0.001), age (p < 0.001) and gender (p < 0.01)Rate of sustained virological response can be determined by factors like rapid virological response and age since they share significant association with one another. More over rate of SVR was more prominent in males than in females. Hepatitis C virus, causative agent of chronic liver disease has affected approximately 175 million people worldwide which make up to almost 3% of the world's populations and 3 to 4 million new cases adds up to this figure annually . Since pDifferent people respond differently to this \u03b1-interferon treatment regimen and various terminologies are used for these responses to therapy. When HCV RNA becomes undetectable (< 50 IU/ml) after treatment completion, the response is characterized as end of treatment response (ETR), treatment response is said to be sustained virological response (SVR) if HCV RNA remain undetectable at the end of follow up (6 months post treatment completion), but if it reappears after 6 months of follow up period than it is termed as relapsed. However patients with a detectable HCV RNA throughout the treatment are non responders (NR) ,8. A 2 lVarious viral and host related factors are responsible for such diversity in responses. HCV genotype and the pre-treatment viral load influence the antiviral response rates and this rate is higher for genotype 2 and 3 than for genotype 1 . GenotypThere are studies that have analyzed the rate of response to IFN-\u03b1 plus ribavirin treatment in Pakistan along with various factors that might predict rate of SVR , howeverPatient's demographic data and samples were collected from 203 patients with chronic HCV infection from July 2009 to July 2010 at Ghurki Trust Hospital Lahore. Out of these 203 patients, 152 patients (first group) received standard HCV treatment of Interferon (3 Million units per ml) 3 injections per week and Ribavirin 10 mg/day/kg of the body weight, While 50 patients (second group) received pegylated interferon (PegIFN 80 million units per ml) 1 injection per week and ribavirin (10 mg/day/kg of the body weight). Patients infected by HCV genotypes None-1 and HCV-1 (or with co-infection with HCV-1) received therapy for 24 weeks and 48 weeks respectively. Total 67 patients were excluded from final data analysis of the study since they were either unwilling to participate in the study at some stage, sara samples were insufficient or lacked the requested information regarding demographic characteristics. All the 136 eligible patients were anti-HCV ELISA and serum HCV RNA positive.Serum samples were collected from enrolled patients before treatment, during treatment (at 4 weeks and 12 weeks) and at the end of treatment. All the serum samples were analyzed for quantitative detection of HCV RNA titer using SmartCycler II real time .HCV genotyping was done on pre-treatment sera samples using the genotyping method described previously .Patient's viral load was evaluated before treatment at 1 month, 3 months, 6 months (non-1 genotypes), 12 months (HCV-1) and at month 6 post treatment. Undetectable HCV RNA titer (< 250 IU/ml) after first month and third month of treatment was described as RVR and EVR response respectively. Undetectable HCV RNA at month 3 of treatment but reappearance of HCV RNA at end of treatment was defined as breakthrough response (BTR). HCV RNA less than 250 IU/mL at 6 months of treatment was defined as end of treatment response (ETR). Undetectable HCV RNA (< 250 IU/ml) 6 months after treatment cessation was defined as sustained virological response (SVR). Patients with detectable HCV RNA throughout the treatment were classified as non responders while ETR patients with detectable HCV RNA after 12 months were defined as relapsers.SPSS version 16.0 was used to analyze all variables . Means with standard deviation and percentage frequencies of variables were calculated where needed. Significant association amongst variables was analyzed by calculating p-value using Fisher's Exact t-test. P-value less than 0.05 were indicative of significant association.Figure Table Table Table Interferon (IFN) plus ribavirin is a routine treatment for chronic HCV infection in Pakistan. It comprises 3 MIU given thrice a week along with ribavirin 800 to1200 mg administered per day. Previous data shows that rate of SVR raises to 38-43% with standard IFN \u03b1 plus ribavirin therapy . A modifIn the current study we have checked several host and viral factors on treatment outcome in patients who received both types of therapies. According to the results of our study, rate of SVR was significantly higher (p < 0.01) in patients received pegylated interferon compared to non-pegylated interferon. Despite of this high rate of SVR, most of our patients received standard IFN-\u03b1 plus ribavirin therapy due to cost factor as the pegylated interferon is at least 25 times expensive than non-pegylated interferon therefore is beyond the reach of poor people in underdeveloped country like Pakistan . It has Age is another factor that might determine the treatment response. It has been reported recently that patient age greater than 50 years increases the risk of relapse . Our stuRecently Shin and co-workers (19) has reported a higher base line titer as an independent risk factor for higher relapse rate. In yet another study low pretreatment viral load was pointed out as predictor of the achievement of improved SVR rate as SVR was significantly higher in patients that had baseline viral load less than 200,000 IU/ml . HoweverAnother important prognostic factor of the attainment of SVR is achievement of RVR. A significantly association was seen between RVR and SVR in the current study (p < .001) as our all patients who had RVR had also achieved SVR. Unfortunately the observed RVR rate is very low as only our 3.6% patients had this type of rapid response at week 4 of treatment. The RVR response has already been established as an important predictor of SVR in more than 85% cases .HCV infection progresses slowly in females than in males . Unlike Our study with reference to previous studies confirms younger age and rapid virological response as important factors for the achievement of sustained virological response. Our study further established that male patients have a greater chance of achieving SVR responses.The authors declare that they have no competing interests.MA and MI conceived the study, participated in its design and coordination and gave a critical view of manuscript writing. MA performed and analyzed the results. AH and SB participated in results analysis. SZ provided clinical data of patients and monitored the patients throught the study. BK, SA, IR, LA, AB, SS and MA participated in data analysis. All the authors read and approved the final manuscript."} +{"text": "Nasal nitric oxide (nNO) has a well-established role as an indirect discriminative marker between patients with and without PCD. All referral services expect to confirm this diagnosis in a minority whereas all referred patients face the inconvenience of travel to an expert centre. a triage test (such as nNO) could reduce this inconvenience and increase the efficiency of the service.To evaluate the utility of nNO for risk stratification within an Australian University Hospital ciliary function service.96 patients with unexplained recurrent respiratory tract infections referred May 2009 and July 2011, underwent nasal brush biopsy with examination of ciliary beating by light microscopy, ciliary beat frequency (CBF) measurement using photometry and EM examination of ciliary ultrastructure. Those patients > 5yrs underwent nNO assessment using a single breath exhalation technique prior to nasal brushing. Healthy volunteers were recruited for nNO measurements.96 had ciliary brushing for CBF and EM. 52 were > 5yrs and also had nNO measurements. 13 confirmed PCD patients 21(8) years [mean(SD)] and 39 non-PCD patients 25(18) years plus 57 healthy subjects 27(17) years were included in the analysis. nNO for the PCD group was 51(48) ppb compared to non-PCD 732(318) ppb (p<0.001) and normals group 833(273) ppb (p<0.001). A cutoff of nNO < 300 ppb has a PPV 93%, NPV 100%, sensitivity 100%, specificity 97% for PCD.Compared with Normals and non-PCD, patients with PCD have very low levels of nNO. Australians without PCD have nNO levels similar to other reported series. nNO measurements > 300ppb have excellent negative predictive value and could be used to stratify expensive and distant referral for comprehensive ciliary structure and function analysis."} +{"text": "Arabidopsis thaliana (Arabidopsis). However, no flowering-time repressors have been functionally identified in the model legume Medicago truncatula (Medicago). In this study, phylogenetic analysis of two closely-related MtSVP-like sequences, MtSVP1 and MtSVP2, showed that their predicted proteins clustered together within the eudicot SVP clade. To determine if the MtSVP-like genes have a role in flowering, they were functionally characterized in Medicago and Arabidopsis. Transcripts of both MtSVP genes were abundant and broadly expressed in vegetative tissues but were detected at much lower levels in flowers in Medicago. Over-expression of the MtSVP genes in Arabidopsis resulted in delayed flowering and flowers with many abnormal phenotypes such as leafy sepals, changes to floral organ number and longer pedicels than the wild type. By contrast, in transgenic Medicago, over-expression of MtSVP1 resulted in alterations to flower development, but did not alter flowering time, suggesting that MtSVP1 may not function to repress the transition to flowering in Medicago.The MADS-domain transcription factor SHORT VEGETATIVE PHASE plays a key role as a repressor of the transition to flowering and as a regulator of early floral development in Arabidopsis thaliana (Arabidopsis) and endogenous signals with carbohydrate status also playing a critical role in controlling the transition to flowering (FLOWERING LOCUS T (FT), TWIN SISTER OF FT (TSF), and SUPPRESSOR OF OVER-EXPRESSION OF CONSTANS1 (SOC1) that go on to promote the transition to flowering.Plants precisely regulate the timing of the transition from vegetative to reproductive growth to optimize sexual reproduction and productivity (idopsis) . In ArabArabidopsis, the gene encoding the MADS-domain transcription factor SHORT VEGETATIVE PHASE (AtSVP) is expressed broadly during vegetative development including in leaves and shoot apices and functions as a flowering time repressor (Arabidopsis svp mutants are early flowering (SVP (35S:AtSVP) leads to a late-flowering phenotype (Poncirus trifoliate (PtSVP) (Arabidopsis. In Antirrhinum majus (snapdragon), the SVP-like gene AmINCOMPOSITA (INCO) has developed a more prominent role in floral meristem identity : BARLEY MADS1 (HvBM1), VEGETATIVE TO REPRODUCTIVE TRANSITION GENE 2 (HvVRT2), and HvBM10 (HvVRT2-like) are expressed in vegetative tissues. RNA interference (RNAi) constructs targeting either BM1 or BM10 did not result in any phenotypic abnormalities or change in heading time, but ectopic expression caused floral reversion in transgenic barley plants (Oryza sativa (rice), there are three SVP-like genes: OsMADS22, OsMADS47, and OsMADS55. Knock-down and over-expression lines showed that these genes appear to function in brassinosteroid signalling and shoot development in Prunus persica (peach) resulted in failure to enter dormancy under cold or short-day induction (Actinidia chinensis (kiwifruit), a perennial vine, all of the four SVP-like genes (AcSVP1\u20134) were implicated in bud dormancy and their ectopic expression in Arabidopsis caused floral abnormalities. However, only AcSVP1 and AcSVP3 were able to complement the Arabidopsis svp mutant (While the role of (EgrSVP) , and PonSVP-like sequences were identified in another large and agronomically important plant family, the Fabaceae, but none have been functionally characterized . Medicago offers several advantages including a diploid and relatively modest genome of ~525Mb that has been sequenced, large collections of accessions with variation in flowering time, and extensive mutant populations for forward and reverse genetic screens -photoperiod and vernalization and Medicago FTa1, an orthologue of FT, plays an important role in promoting flowering under these inductive conditions (Medicago lacks flowering-time repressors that belong to the FLC-MADS AFFECTING FLOWERING (MAF) clade in the Brassicaceae family or genes that are similar to the VERNALIZATION2 (VRN2) flowering time repressor from cereals that are directly or indirectly repressed by vernalization and other pathways such as the Arabidopsis autonomous pathway SVP-like genes were identified by BLAST searches against the Medicago EST and BAC sequences. Alignment was performed using ClustalW in the Geneious software package [version 6.0.5 available from www.geneious.com (last accessed 1 November 2013) ]. A phylogenetic tree was generated using the Neighbor\u2013Joining method via bootstrap resampling in the Geneious program. The accession numbers for the sequences used are as follows: Medicago MtSVP1 (Medtr5g032520), MtSVP2 (Medtr5g032150), Medtr5g066180; Arabidopsis AtSVP (At2G22540), AtAP1 (At1g69120), AtSOC1 (At2g45660), AtAGL14 (At4g11880), AtAGL19 (At4g22950), AtAGL24 (At4g24540); rice OsMADS22 (AB107957), OsMADS47 (AY345221), OsMADS55 (AY345223); barley HvBM10 (EF043040), HvVRT2 (DQ201168), HvBM1 (AJ249142); Lolium perenne (ryegrass) LpMADS10 (AAZ17549); Euphorbia esula (leafy spurge) EeDAM2 (ABY60423); peach PpDAM1 (DQ863253), PpDAM2 (DQ863255), PpDAM3 (DQ863256), PpDAM4 (DQ863250), PpDAM5 (DQ863251), PpDAM6 (DQ863252); Brassica campestris (Chinese cabbage) BcSVP (DQ922945); snapdragon AmINCOMPOSITA (CAG27846); Eucalyptus grandis EgrSVP (AY263809); Populus trichocarpa (poplar) PtSVP (XM_002310274); Lotus japonicus LjSVP (TC40194); Pisum sativum (garden pea) PsSVP (AY830919); Glycine max (soybean) GmSVP1 (Glyma01g02880), GmSVP2 (Glyma02g04710), GmSVP3 (Glyma06g10020), GmSVP4 (Glyma07g30040), GmSVP5 (Glyma08g07260), GmSVP6 (Glyma13g33040), GmSVP7 (Glyma15g06300), GmSVP8 (Glyma15g06310); kiwifruit AcSVP1 (JF838216), AcSVP2 (JF838217), AcSVP3 (JF838218), AcSVP4 (JF838219); and Phaseolus vulgaris (common bean) PvSVP (TC35086). These sequences were obtained from previous studies (The two Medicago genotypes used in this study. They belong to the two subspecies of Medicago truncatula Gaertn (barrel medic), ssp. truncatula and ssp. tricycla, respectively. The reverse genetic lines for MtSVP2 were obtained from the Samuel Roberts Noble Foundation . Scarification, germination, seed vernalization prior to growth in LD conditions (16/8h light/dark), and cultivation of Medicago plants were done as described previously are the two Arabidopsis wild type (WT) Columbia (Col) and svp-41 mutant in the Col background were grown in LD. The svp-41 mutant harbours a deletion of bases 193 and 194 in the AtSVP coding sequence (svp-41 mutant to flower earlier than WT Col plants.The PROTODERMAL FACTOR 2 (PDF2) or TUBULIN. The primers used are shown in Supplementary Table S1 at JXB online.RNA extraction, cDNA synthesis, and qRT-PCR were performed as previously described (MtSVP overexpression (35S:MtSVP) constructs, the cDNAs of MtSVP1 and MtSVP2 were amplified using the Phusion Hot Start High-Fidelity DNA Polymerase (New England Biolabs) with gene-specific primers (Supplementary Table S1 at JXB online) and then cloned in pCR8/GW/TOPO TA (TOPO) entry vector . The entry clones were linearized with 5U XhoI and then recombined into the plant transformation vector, pB2GW7 according to the manufacturer\u2019s instructions. Agrobacterium tumefaciens strain GV3101 with expression clones was applied to Arabidopsis WT Col and svp-41 mutant flower buds as previously described by \u20131).In order to generate Medicago R108 and regeneration via somatic embryogenesis was conducted as previously described by 35S:MtSVP1 or vector only (pB2GW7) as a control. The control group went through somatic embryogenesis on selective or non-selective medium as the negative control or the regeneration control, respectively. The transformant plants were selected with 1\u20132mg l\u20131 of dl-phosphinothricin (Sigma). Regenerated T0 transformed plantlets were transferred to soil and sprayed again with 120mg l\u20131 Basta (Kiwicare) to confirm their transgenic nature. PCR-genotyping also confirmed the presence of the transgenes.Transformation of Tnt1 insertions at the MtSVP genes were screened using a reverse genetic approach conditions. Their gDNA was extracted and the nature and direction of Tnt1 insertion in the lines were confirmed using PCR. The expression levels of MtSVP2 were measured in the transgenic homozygous plants from the two lines using qRT-PCR. All the primers used are shown in Supplementary Table S1 at JXB online.Mutant plants with Medicago genomic and EST sequences using BLAST analyses with the AtSVP protein were two closely-related SVP-like genes named MtSVP1 and MtSVP2. MtSVP1 (Medtr5g032520) (MtSVP2 (Medtr5g032150) encode MADS-domain proteins with 89% aa identity to each other and 70% and 66% aa identity respectively, with the AtSVP protein. MtSVP1 and MtSVP2 were less similar to AGL24, which is the most closely related MADS domain protein to SVP consistent with the features shared by the MADS-domain family of transcription factors in plants Medicago wild type (WT) plants was compared was used as a marker of the transition to flowering .More marked differences in gene expression were observed between the two genes in their developmental pattern of expression in apical buds. flowers . MtSVP2 flowers . While aMtSVP genes in flowering-time control and the regulation of flower development, they were overexpressed in the heterologous plant Arabidopsis. cDNA clones of the MtSVP genes were generated from total RNA of Jester wild-type plants, fused to the cauliflower mosaic virus (CaMV) 35S promoter (35S:MtSVP1 and 35S:MtSVP2) and introduced into Arabidopsis WT Col plants. Nine T135S:MtSVP1 and 35S:MtSVP2 plants with single locus insertions were identified (see Supplementary Table S2A at JXB online).To assess the potential roles of the two 2 progeny of eight of these transgenic lines had delayed flowering compared with WT Col plants (Supplementary Table S2 at JXB online). At the onset of flower formation, the transgenic plants were easily distinguished from WT Col , which have recently revealed that AtSVP directly regulates the expression of a large number of Arabidopsis genes implicated in many different plant processes used in this study.Supplementary Table S2. Flowering time of T2 transgenic plants from the overexpression of MtSVP1 and MtSVP2 in WT Col under LD conditions and classification of floral phenotypes."} +{"text": "Previous studies demonstrated that the collapsibility index (percent decrease in inferior vena cava (IVC) diameter with inspiration) of > 50% and an IVC/Aorta ratio of < 0.8 correlated with a low intravascular volume.Our study sought to determine if bedside ultrasound (BUS) measurements of the IVC diameter correlate with central venous pressure (CVP) measurements as an indicator of intravascular volume status in acutely ill children.A convenience sample of children < 21 years-old who were admitted to the pediatric critical care unit and required CVP monitoring had BUS measurements of both IVC and aortic diameters with simultaneous CVP measurement. The collapsibility index and IVC/Aorta ratio (transverse view) were calculated from these measurements. A CVP \u2264 8 mmHg was considered as a marker for decreased intravascular volume.Of the 51 participants, 21 (43%) had a CVP < 8 mmHg. Eight of 51 (16%) children had a collapsibility index > 50% and 8 of 43 (18%) had an IVC/Aorta ratio of < 0.8. The sensitivity of a collapsibility index \u2265 0.5 to predict a CVP \u2264 8 mmHg was 14%, the specificity was 83%, the positive predictive value was 38% and the negative predictive value was 57%. Neither collapsibility index nor IVC/Aorta correlated with CVP in assessing intravascular volume in our study population.Based on these data, the IVC and aortic measurements by BUS are not reliable indicators of intravascular volume (as determined by CVP) in acutely ill children."} +{"text": "Cardiovascular magnetic resonance (CMR) is the gold standard to measure right ventricular ejection fraction (RVEF). Longitudinal shortening is historically known to be the predominant part of its global systolic function and less attention has been paid to the transversal contraction. The aim of this study was to evaluate RV transverse motion in a large cohort of patients referred for CMR and assess its relationship with RVEF.We retrospectively analyzed the CMR scans of 300 consecutive patients referred for CMR between January and December 2010. Reference RV ejection fraction was determined from short axis sequences using the volumetric method. Transverse parameters called RV fractional diameter changes (FDC) were calculated after measuring RV diastolic (D) and systolic (S) diameters at basal and medium level in short axis view (respectively FBDC and FMDC) FBDC= 100 x (D-S)/D / 51.32 = 35.7%).We also measured the tricuspid annular plane systolic excursion (TAPSE) in the four chambers view as a longitudinal reference.Population was divided into 2 groups according RVEF, 250 patients had a preserved RVEF (>40%) and 50 had a RV dysfunction (RVEF \u226440%). Both transverse and longitudinal motions were significantly reduced in the group with RV dysfunction (p<.0001). After ROC analysis, areas under the curve for FBDC ,FMDC and TAPSE, were respectively 0.79, 0.82 and 0.72 with the highest sensitivity and specificity of 68% and 88% for FMDC ( threshold at 19.9%) to predict RVEF. FMDC had an excellent negative predictive value of 93%.Right ventricular fractional diameter changes, especially at the medium level, appear to be accurate for semi quantitative assessment of RV function by CMR.No funding."} +{"text": "The objectives of this special issue on paraoxonases (PONs) are to bring the latest aspects of paraoxonases research on the genetics, biochemistry, cell biology, and structural biology of PONs. The issue also addresses the role of PONs in human diseases . Inflammatory and oxidative stress-related diseases, such as diabetes or rheumatoid arthritis are also favorably affected by PON. PONs also provide microbial protection by hydrolyzing bacterial quorum lactone. The pivotal role of the PON family in a variety of inflammatory diseases, and in preventing the toxicity of organophosphorus insecticides and nerve agents, has made PONs an interesting target for both clinicians and scientists alike. Research into the paraoxonase family of enzymes has increased dramatically, especially following the initiation of the paraoxonase conferences. Five international conferences on paraoxonases were organized between 2004 and 2012, where the PON scientific community gathered to discuss PONs research achievements and future scientific directions. These meetings took place in Ann Arbor (MI), Debrecen (Hungary), Los Angeles (CA), La Pineda (Spain), and Columbus (OH). According to PubMed, only few PON papers were published before 1980. From 1980 till these days, about 3000 papers were published.\u03b2 propeller protein structure, and hydrolyze esterase/lactonase activities.The PONs gene cluster contains three adjacent gene members, and all of the three PON genes share high sequence, indentify a similar PONs play a clear protective role against cardiovascular diseases. Major cardioprotective PON characteristics include, beside their potent antioxidant properties, the following: for PON 1, favorable effects on macrophage cholesterol metabolism, for PON 2, attenuation of macrophage triglyceride accumulation, and for PON 3, improvement in bile acids metabolism.Human serum HDL-associated paraoxonase (PON1) is an esterase that possesses cardiovascular protective properties which result in the following antiatherogenic functions: (1) attenuated oxidative stress in serum, in lipoproteins, in macrophages, and in atherosclerotic lesions; (2) decreased oxidized LDL uptake by macrophages; (3) inhibited macrophage cholesterol biosynthesis rate; (4) stimulated HDL-mediated cholesterol efflux from macrophages.Major PON1 inactivators include (1) oxidative stress ; (2) high cholesterol ; (3) high triglycerides ; (4) high glucose .Macrophage PON2 regulation (as related to atherogenesis) differs from that of serum PON 1 in the following characteristics: (1) stimulation (not inhibition as shown for PON1) by oxidative stress and also by anti oxidants (like PON1); (2) stimulation by high glucose (PON1 is inhibited) and also by insulin (like PON1); (3) stimulation by urokinase plasminogen activator (uPA); (4) stimulation by high triglycerides (PON1 is inhibited); (5) stimulation by arachidonic acid and also by its product prostaglandin E2 (like PON1); (6) inhibition by high cholesterol and reversal effect by statins (like PON1).Michael AviramMichael Aviram"} +{"text": "Balance is a complex process involving visual, vestibular and neuromuscular control. Migraineurs often report vertigo and dizziness symptoms during and post migraine. The Biodex Balance System SD 1 is a reliable method to measure dynamic balance. Little research 2 has examined dynamic balance in migraineurs compared to individuals who do not have migraines.The purpose of this research is to examine the differences between migraineurs and controls dynamic balance at two testing intervals: baseline (migraine free 7 days) and post migraine (within 48 hours of migraine onset).20 controls (C) (age 25.70\u00b1 10.77) and 17 Migraineurs (M) (age 25.24\u00b1 9.55) completed dual limb support testing on the Biodex Balance System SD. Limits of Stability (LOS) testing at moderate skill level (75%) involved center of gravity control within their base of support. The clinical test of sensory integration and balance tested stability and sway indexes within four conditions (eyes open/closed on firm vs foam surface) for 30 second intervals.A repeated measures ANOVA revealed significant differences between migraineurs and non-migraineurs in overall LOS post migraine. Significant decreases were found between shift of balance to the right and balance to the left post migraine. No significant differences were found between groups for overall LOS at baseline (p=.703). No significant differences were found between groups for stability or sway indexes for all conditions on the clinical test of sensory integration.Migraineurs exhibit difficulty with center of gravity shifts to the right and left and overall dynamic LOS post migraine. Once LOS is exceeded a fall, stumble or step will ensue. This suggests decreases in lower extremity strength, proprioception and vestibular deficiencies."} +{"text": "To investigate whether five treatment sessions of cupping massage (CM) improve chronic non-specific neck pain (cNP) and influence pain related neurophysiologic measures.50 patients with cNP (age 53.3 \u00b1 10.4 yrs) were examined at baseline and then randomized to treatment (TG) or standard medical care (SMC). Data were collected at baseline (T1) and T2 (for TG after treatment). TG received five treatment sessions of CM, while the SMC group was waiting. Subjective outcome measures included visual analogue scale for pain (VAS), Neck Pain Disability Index (NDI), and quality of life (SF36). Neurophysiologic testing included Quantitative Sensory Testing subtests mechanical detection threshold (MDT), pressure pain threshold (PPT), vibration threshold (VDT) and two-point discrimination (2PD) at the point of maximum pain (Pmax), close to maximum pain (Pclose), and hand and foot as control areas. ANCOVA analyses were conducted across post-pre differences with baseline values as covariates.CM reduced pain and NDI scores in TG, while SMC showed no change . \u201cPhysical health\u201d (SF36) increased . Neurophysiological data: TG showed an increase for PPT at Pmax .CM improved pain and increased subjective well being in cNP patients. The increase in PPT is consistent with a reduced peripheral sensitization of deeper tissues and supports previous findings on dry cupping in chronic cNP ."} +{"text": "Medical Parasitology subject is one of basic medical science. We have integrated subject through community based learning.Assess medical students\u2019 attitudes toward practice in laboratory (TC) parasitological examination in the rural community (MPEC).Cross-sectional descriptive study was constructed among 46 medical students during April to July 2012. Attitudes were compared between practice in laboratory (TC) and mobile parasitological examination in the community (MPEC).22 Males and 24 Females) medical students. Most of students was highly satisfied with MPEC. Student\u2019 skill, they could be identified parasites during community studied. A total of 85 stool sample was examined and found 7 samples were infected with hookworm (5 patients), Strongyloides stercolaris (1 patient) and Taenia sp. (1 patient), respectively.A total of 46 (That modification in educational methods. MPEC experience in particular can favorably influence students' attitudes toward basic sciences.None declared"} +{"text": "The objective of the study is to investigate whether PVI, a non-invasive and continuous tool, can predict fluid responsiveness in mechanically ventilated patients with circulatory insufficiency.In patients with acute circulatory failure related to sepsis or hypovolemia, volume expansion is used as first-line therapy in an attempt to improve cardiac output. Dynamic indices based on cardiopulmonary interactions and variation in left ventricular stroke volume like respiratory variations in arterial pulse pressure (\u0394PP) are able to predict response to fluid loading in mechanically ventilated patients. The Pleth Variability Index (PVI) (Masimo2/FIO2 < 100 mmHg); contraindication for passive leg raising (PLR); altered left ventricular ejection fraction; hemodynamic instability during the procedure. We performed fluid challenge with 500 ml of 130/0.4 hydroxyethylstarch if \u0394PP \u226513% or with PLR otherwise. PVI, \u0394PP and cardiac output (CO) estimated by echocardiography were recorded before and after fluid challenge. Fluid responsiveness was defined as an increase in CO \u226515%.A prospective study in a surgical ICU of a university hospital. Forty mechanically ventilated patients with circulatory insufficiency were included in whom volume expansion was planned by the attending physician. Exclusion criteria included spontaneous respiratory activity; cardiac arrhythmia; known intracardiac shunt; severe hypoxemia with percentage changes in CO (\u0394CO) induced by fluid challenge, suggesting the higher PVI at baseline, the higher \u0394CO after volume expansion.Twenty-one patients were responders and 19 were non-responders. Median (interquartile range) PVI (26% (20 to 34%) vs. 10% (9 to 14%)) and \u0394PP (20% (15 to 29%) vs. 5% (3 to 7%)) values at baseline were significantly higher in responders than in nonresponders. A PVI threshold value of 17% allowed discrimination between responders and nonresponders with a sensitivity of 95% (95% CI = 74 to 100%) and a specificity of 91% (95% CI = 70 to 99%). PVI at baseline correlated (PVI can predict fluid responsiveness non-invasively in ICU patients under mechanical ventilation."} +{"text": "Besides confirming direct regulation of previously implicated Zic3 targets of the Nodal and canonical Wnt pathways, analysis of gastrula stage embryos uncovered a number of novel candidate target genes, among which were members of the non-canonical Wnt pathway and the neural pre-pattern genes. A similar analysis in zic3-expressing cells obtained by FACS at segmentation stage revealed a dramatic shift in Zic3 binding site locations and identified an entirely distinct set of target genes associated with later developmental functions such as neural development. We demonstrate cis-regulation of several of these target genes by Zic3 using in vivo enhancer assay. Analysis of Zic3 binding sites revealed a distribution biased towards distal intergenic regions, indicative of a long distance regulatory mechanism; some of these binding sites are highly conserved during evolution and act as functional enhancers. This demonstrated that Zic3 regulation of developmental genes is achieved predominantly through long distance regulatory mechanism and revealed that developmental transitions could be accompanied by dramatic changes in regulatory landscape.Zic3 regulates early embryonic patterning in vertebrates. Loss of Zic3 function is known to disrupt gastrulation, left-right patterning, and neurogenesis. However, molecular events downstream of this transcription factor are poorly characterized. Here we use the zebrafish as a model to study the developmental role of Zic3 The Zic3 transcription factor regulates early embryonic patterning, and the loss of its function leads to defects in left-right body asymmetry. Previous studies have only identified a small number of Zic3 targets, which renders the molecular mechanism underlying its activity insufficiently understood. Utilizing two genomics technologies, next generation sequencing and microarray, we profile the genome-wide binding sites of Zic3 and identified its target genes in the developing zebrafish embryo. Our results show that Zic3 regulates its target genes predominantly through regulatory elements located far from promoters. Among the targets of Zic3 are the Nodal and Wnt pathways known to regulate gastrulation and left-right body asymmetry, as well as neural pre-pattern genes regulating proliferation of neural progenitors. Using enhancer activity assay, we further show that genomic regions bound by Zic3 function as enhancers. Our study provides a genome-wide view of the regulatory landscape of Zic3 and its changes during vertebrate development. The Zic family of transcription factors (TFs) is involved in such process situs inversus) in humans Xenopus, Zic3 established left-sided expression of Xnr1 and Pitx2zic3 is expressed symmetrically along the L-R axis in the Xenopus embryo and its loss-of-function (LOF) affects structures in which its expression was not detected Of particular interest is ZIC3, which is linked to the heritable defects of the left-right internal organs placement and blastoderm margin does not express GFP. This suggests that a small fraction of non-neuronal zic3-expressing cells may be present in the GFP-negative pool of cells.The earliest at 3 hpf , coincidastoderm , and is astoderm . To captnal cord . To idenpression , we consSequencing of the 8 hpf ChIP sample generated 23,945,552 reads ; the 24 hpf ChIP sample generated 23,083,504 reads . We identified 3209 and 2088 Zic3 binding sites (hereafter referred to as peaks) with high significance value at 8 hpf and 24 hTo determine the biological relevance of our data, we used the gene association rule \u2018basal plus 100 kb extension\u2019 according to GREAT algorithm To identify the common regions bound by Zic3 as well as those unique to either developmental stage, we overlapped the 8 hpf and 24 hpf peaks . Taking de novo motif search using sequences within 50 bp of the top 1000 peaks summit. The highest scoring motif in both datasets consisted of a CAGCAG core -mediated knockdown. Embryos injected with the same MO dosage as in Cast oep, lft1 and pitx2 , were concurrently upregulated in Zic3 morphants without association with peaks has changed in Zic3 morphants based on microarray data. This implied that such genes could be the indirect targets of Zic3. Such observation provided further support for Zic3 regulation of the canonical Wnt pathway. The inhibition of canonical Wnt signaling by Zic3 was previously reported in frogs as a mechanism for organizer development We identified 1316 genes differentially expressed in MO injected embryos . These genes were co-expressed with zic3 during gastrulation . In addition, other genes with similar function, such as ccdc88a (probe generated from BC057440 which correspond to the annotated ccdc88a sequence) tsg101Several genes implicated in cell migration and polarity were among the targets. These include zic3 targets during late neurogenesis, we performed microarray expression analysis on 24 hpf GFP-positive zic3 expressing cells that were FACS-sorted (zic3 expression), we identified genes enriched in GFP-positive cells (zic3-expressing cells). A total of 689 genes were enriched in zic3-expressing cells (zic3-coexpressed genes). Among these genes were six members of the Zic family and other genes expressed in the dorsal neural tube. This confirmed the identity of the sorted cells as dorsal neural cells. Among the zic3-coexpressed genes, 167 had at least one peak within 100 kb of their TSS, rendering them putative Zic3 targets were found under this category, which may reflect that the zic3-expressing cells in the dorsal neural tube are not differentiating. Interestingly, the presence of a Zic3 peak in association with oep suggests that a similar inhibition of Nodal by Zic3 occurs at both 8 hpf and 24 hpf , but none of these were co-expressed with zic3 at 8 hpf, while only 6 had expression overlapping with zic3 at 24 hpf (not shown).The large number of Zic3 binding sites in the distant intergenic regions suggested that Zic3 may direct the expression of target genes by binding to the distal regulatory elements. In support of this idea, relevant biological categories could be observed among genes associated with peaks located outside of their basal regions of \u22125 kb to +1 kb of TSS , axin1 , lft1 , dvl2 , and invs . A canonical Zic3 motif was present within 100 bp of each peak summit except for fragment 10-02. Only fragment 16-297, associated with invs, showed enhancer activity , axin2 , and pitx2 . These peaks had at least one canonical Zic3 motif and exhibited positive enhancer activity d humans , with 31d humans . Genes ad humans .gfp expression at either 8 hpf or 24 hpf, or both . Of these eight, four continuously drove reproducible tissue-specific gfp expression in various regions of the CNS up to 24 hpf drove or both . Of theso 24 hpf , which opf drove higher at 8 hpf . Togethein vivo.The majority of Zic3 binding sites were found outside promoter regions. While this could be partially attributed to the incomplete annotation of promoter regions in the zebrafish genome, the predominantly distal distribution of Zic3-binding sites revealed that Zic3 regulates transcription largely via distal regulatory elements. Such distribution of binding sites was previously observed in other genome-wide analyses of several TFs in cell culture or mammalian tissues Some of the Zic3 binding sites overlapped with CNEs, most of which drove expression in neural tissues. CNEs are known to regulate developmental genes in vivo. For instance, the concurrent upregulation and downregulation of different subsets of direct target genes by Zic3 suggest that Zic3 binding can result in either activation or repression of target genes, and implies that additional mechanisms determine these two outcomes. Another facet of the data revealed distinct Zic3 binding profiles at 8 hpf and 24 hpf. The genes associated with binding events at these two stages showed relevant functional enrichments. This shift in binding was not dictated by a change in DNA recognition motif as almost identical dominant motifs were identified in both stages.Cell culture studies have demonstrated interactions between multiple enhancer elements in regulating the transcription of a target gene The combinatorial analysis of ChIP-seq and microarray datasets revealed an entirely distinct set of candidate Zic3 target genes at 8 hpf and 24 hpf. Whereas not totally unexpected, this analysis revealed some surprises. First, a developmental switch towards regulation of different members within the same signaling pathway was detected. In the context of Wnt signaling this shifted Zic3 impact from the intracellular part of Wnt signaling towards extracellular ligands in this pathway. Second, that cells expressing Zic3 show a reduced level of transcription of proneural genes placed an impact of Zic3 on cells that are in a state either before or after neural differentiation. Zic3 has been linked with pluripotency of stem cells in mammals DrosophilaOne implication of an interactive regulatory landscape is that genes targeted by a particular TF may not be determined by simply observing binding of the TF near its genomic locus. Additional proof, such as responsiveness of the particular target gene to LOF of the TF, would be necessary. In our data, there is a surplus of Zic3 binding events compared to those associated with responsive target genes. Widespread binding of TFs exceeding their known target genes have been reported in cell culture and in in vitro data have demonstrated physical and functional interactions between Zic and Gli proteins in vivo in regulating transcription of target genes.The highly interconnected TF regulatory network also necessitates a careful interpretation of enhancer function by reporter assays: while such assays can be useful to identify independently acting regulatory elements, evidence exists for regulatory elements acting in tandem, resulting in higher transcriptional output per se could have been sufficient to ensure a proper L-R asymmetry. However, our results suggested that Zic3 may also regulate the non-canonical Wnt (PCP) signaling pathway which is implicated in ciliogenesis. Interaction of these signaling pathways culminates in the establishment of a proper embryonic L-R axis mkks was also found as one of the Zic3 targets which is implicated in mesendodermal specification targets which isneurog1 expression was upregulated upon Zic3 knockdown, and Zic3 binding sites were found near neurog1, as well as other proneural genes such as neurod4 and ncam1a, Zic3 may have an additional direct role in neural differentiation as its inhibitor. This possibility is also supported by the downregulation of her9. This places Zic3 within a regulatory landscape of Notch signaling in support of an early hypothesis based on functional analysis of Zic1 Our finding that Zic3 regulates genes implicated in proliferation of neural progenitors agrees with the idea that Zic3 has properties of a stem cell factor Zebrafish of wild type (AB strain) and transgenic line SqET33 Dechorionated 24 hpf transgenic embryos were deyolked in PBS by pipetting through the 1 ml pipette tip. Cells were dissociated with trypsin solution (0.05% trypsin and 0.2 mM EDTA) in PBS for 15 min at room temperature. To facilitate dissociation of cells, embryos were pipetted through the 200 \u00b5l pipette tip. Trypsin was inhibited with complete protease inhibitor cocktail (Roche) and cell suspension was filtered through a nylon mesh . Immediately, an equal volume of 4% paraformaldehyde (PFA) in PBS was added to cell suspension and cells were fixed for 10 min at room temperature. Reaction was stopped by an equal volume of ice-cold 0.25 M glycine in PBS, cells were washed three times with 0.125 M glycine-PBS and resuspended in the same buffer. Cell sorting was carried out with FACSAriaII Cell Sorter (BD Bioscience). To set autofluorescence level, cell sorter was calibrated with PFA-fixed GFP-negative cells before cell separation. GFP-positive and GFP-negative cells were collected in 0.125 M glycine-PBS, frozen in liquid nitrogen and kept at \u221280\u00b0C until use. For microarray analysis, PFA fixation step was omitted and cells were sorted into complete L-15 Leibovitz medium (Gibco) containing 20% fetal bovine serum.de novo motif finder Tetraodon CNE database to take into consideration the genome duplication event during teleosts evolution, which relaxed selection pressure on the conservation of important developmental enhancers Tetraodon tetNig2 assemblies. A threshold of at least 70% sequence conservation within every 50 bp was used to define CNEs in each species.Chromatin Immunoprecipitation (ChIP) was performed according to an established protocol with an addition of a deyolking step according to Link and colleagues (2006), with modifications see . ChIP DN5\u2032-GGG GAC AAG TTT GTA CAA AAA AGC AGG CTT CGA AAA CCT GTA TTT TCA GGG CAG CTT ACG TGA AAT TGC G CTC-3\u2032 and 5\u2032-GGG GAC CAC TTT GTA CAA GAA AGC TGG GTT TAC TCC ACC TGA AAA CGG ACT TG-3\u2032; Zic3_ZF2-5: 5\u2032-GGG GAC AAG TTT GTA CAA AAA AGC AGG CTT CGA AAA CCT GTA TTT TCA GGG CGC CTT CTT CAG ATA CAT GCG-3\u2032 and 5\u2032-GGG GAC CAC TTT GTA CAA GAA AGC TGG GTT TAT GAT TCG TGT ACC TTC ATA TG-3\u2032. Each forward and reverse primer contained an attB recombination site overhang, with an additional Tobacco Etch Virus (TEV) protease cleavage site in the forward primer preceding the N-terminal Zic3 coding sequence. Protein expression and purification was performed as previously described . Electrophoretic mobility shift assay (EMSA) was performed as previously described st BASE, Singapore) were annealed by heating to 95\u00b0C for 5 minutes in annealing buffer and left in room temperature to cool down overnight. These were subsequently incubated with the recombinant Zic3 in EMSA buffer for 1 hour at 4\u00b0C. The reaction was subsequently run on 5% native Tris-Glycine polyacrylamide gel electrophoresis. Gel was scanned in Typhoon Scanner . The affinity of protein to DNA was determined by titrating 0\u2013250 nM of protein against 1 nM of annealed probes.Two recombinant constructs of the zebrafish Zic3 protein were produced, the full-length protein (Zic3_ORF) and the DNA-binding domain encompassing Zn-fingers 2 to 5 . DNA sequences corresponding to each domains were PCR-amplified using the following primers: Zic3_ORF: 5\u2032-AGG TTA GTG GAG TGA ACG GGT ACC G-3\u2032. A standard control antisense MO was also obtained from Gene Tools, LLC with the following sequence 5\u2032-CCT CTT ACC TCA GTT ACA ATT TAT A-3\u2032. For microarray, 1.7 ng Zic3 MO was injected into 1-cell stage embryos. Rescue was performed using 20 pg of zic3 mRNA without morpholino-binding site. Capped zic3 mRNA was synthesized using mMessage mMachine Kit . Results were obtained from at least three different experiments on embryos from random pairs.Zic3 knockdown was performed using a translation-blocking antisense morpholino oligonucleotide (MO) purchased from Gene Tools, LLC (USA). The MO sequence was http://123.136.65.67/). Genes were subjected to pathway assembly using Ingenuity Pathway Analysis . Selected genes containing 44,000 oligonucleotide probes was used. The microarray was performed according to Agilent's One-Color Microarray Based Gene Expression Analysis (Quick Amp Labeling) protocol and RNA Spike-In-One Color. Arrays were probed using cDNAs reverse transcribed in the presence of Cy3-dUTP using 600 ng of total RNA from either wild-type control or Zic3 knockdown embryos (8 hpf), or from either non zic3-expressing cells or zic3-expressing cells (24 hpf). Labeled cDNA was denatured and hybridized at 42\u00b0C for 16 h in a hybridization oven . After hybridization, the slides were washed and scanned for fluorescence detection on Agilent DNA Microarray Scanner. Scanned images were analyzed using Agilent Feature Extraction Software (v10.5.1.1). Feature extracted data were analyzed in Genespring software . Statistically significant gene expression was identified using Significance Analysis of Microarrays (SAM 3.05) for each successive time point ed genes were valSalI and BamHI sites of the pTol2-GFP reporter vector containing a minimal promoter from the mouse cFos gene egfp expression at 24 hpf. A consistent egfp expression pattern observed in at least 20% of injected embryos was considered as positive. The reporter vector alone showed expression in muscles and blood cells in G0 embryos (data not shown). Embryos positive for egfp expression were subsequently processed for whole mount immunohistochemistry (IHC) with anti-GFP antibody. qPCR was used to determine egfp expression level at 8 hpf since morphological identification of tissue specificity at this stage was difficult.Tested genomic regions encompassing the peaks with \u223c200 bp flanking sequence at each side were amplified using PCR and cloned into Figure S1Anti-Zic3 antibody specifically recognizes recombinant and endogenous Zic3 proteins. A, Alignment of C-terminal amino acid sequences of zebrafish Zic proteins. Anti Zic3 antibody was designed to recognize a region in the C-terminal of Zic3 protein which is unique, having between 15% to 38% similarity with other Zic family members (highest similarity with Zic2a and Zic4). B, full-length Zic3 recombinant protein containing an MBP tag (Zic3_ORF) was expressed in bacteria and isolated with affinity chromatography. Left panel shows protein SDS-PAGE of uninduced bacterial lysate containing Zic3 expression construct (U), induced bacterial lysate expressing Zic3_ORF (I), eluted lysate of Zic3_ORF (E), and Zic3_ORF treated with TEV protease to remove MBP tag (TEV). Right panel shows Western blot with anti Zic3 primary antibody. Additional lanes show Coomassie blue staining and western blot of a duplicate gel of whole embryonic cell extract (wce), as well as western blot of immunoprecipitated Zic3 protein (ChIP) detected using anti Zic3 primary antibody and a light chain specific anti rabbit IgG secondary antibody. Notice protein bands corresponding to Zic3 with MBP tag and without tag , as well as IgG light chain at 25 KDa. C, mass spectrometry result of the recombinant protein band at 52 kDa , which matched to zebrafish Zic3 sequence.(TIF)Click here for additional data file.Figure S2Functional categories enriched in different subsets of genes associated with peaks identified at 8 and 24 hpf. List of biological process and tissue specific expression terms among genes associated with peaks found at both 8 hpf and 24 hpf (Class I), at 8 hpf only (Class II), and at 24 hpf only (Class III). Light and dark grey bars represent the expected and observed enrichments, respectively, of functional categories indicated along the y-axis. Analysis performed with DAVID algorithm.(TIF)Click here for additional data file.Figure S3EMSA of Zic3 motifs. A, hierarchical clustering of several motif identified from ChIP-seq as well as other motifs known to interact with Zic3. B, EMSA performed using the mouse Zic3 recombinant protein (TIF)Click here for additional data file.Figure S4sox17a expression shows the expansion of the mesendoderm territory (marked by dashed line) in Zic3 morphants. B, WISH of ntla in the notochord at 8 hpf in control and Zic3 morphants. C\u2013D, WISH of pax3a and sox19a at 10 hpf to label the neural plate border. Dashed line marks the width of neural plate. E\u2013F, average width of the neural plate was measured at the dashed line in control and Zic3 morphants. Error bars represent standard deviation.Defects in cell fate specification and convergent-extension movement in Zic3 knockdown embryos. A, WISH of (TIF)Click here for additional data file.Figure S5rock2b shows the reduction of expression domain in the dorsal forerunner cells in Zic3 morphant embryos. B, WISH of dvl2 in control and Zic3 morphant embryos. C, WISH of representative Zic3 target genes at 8 hpf whose expression were downregulated in Zic3 knockdown embryos. Dorsal is to the right.WISH of Zic3 target genes. A, WISH of (TIF)Click here for additional data file.Figure S6Effect of Zic3 knockdown in migration of the dorsal forerunner cells. Expression of sox17a (A\u2013B) and foxj1a (C\u2013D) marks the dorsal forerunner cells in controls and Zic3 morphant embryos at 8 hpf.(TIF)Click here for additional data file.Figure S7Enrichment of functional categories of genes associated with proximal and distal Zic3 binding sites. Light and dark grey bars represent the expected and observed enrichments, respectively, of functional categories indicated along the y-axis. A, list of biological processes enriched in genes with peaks beyond \u22125 kb to +1 kb region. B, a similar enrichment in genes with peaks beyond +/\u221250 kb from TSS. Analysis performed on DAVID algorithm.(TIF)Click here for additional data file.Figure S8Zic3 is responsible for reporter gene expression in tested enhancers. Enhancer fragments 7-211, 4-16, and 17-24 was either injected alone or co-injected with Zic3 MO into wild-type embryos. GFP expression level in the MO co-injected constructs (MO) was assayed using qRT-PCR and presented as a fraction of that driven by the construct alone (control). A significant decrease in reporter expression when construct was co-injected with MO confirms Zic3-dependent activation of reporter expression through binding to these enhancers.(TIF)Click here for additional data file.Table S1List of validated ChIP-seq peaks of Zic3 binding. Genomic location of some ChIP-seq peaks validated by qPCR including sequences of primer used for qPCR.(XLS)Click here for additional data file.Table S2GO enrichment of ChIP-seq peaks at 8 hpf. Analysis was performed using the DAVID algorithm.(XLS)Click here for additional data file.Table S3List of genes detected by microarray analysis (8 hpf). These genes were differentially regulated according to microarray analysis of transcriptome in Zic3 morphants and control embryos at 8 hpf.(XLS)Click here for additional data file.Table S4GO enrichment of microarray genes (8 hpf).(XLS)Click here for additional data file.Table S5Genes with ChIP-seq peak at 8 hpf. List of genes expressed differentially at 8 hpf with at least one ChIP-seq peak within 100 kb of their TSS.(XLS)Click here for additional data file.Table S6GO enrichment of microarray genes with ChIP-seq peak at 8 hpf. List of enriched GO terms among genes differentially regulated in microarray and associated with at least one peak in ChIP-seq (Zic3 target genes) at 8 hpf.(XLS)Click here for additional data file.Table S7qRT-PCR of selected target genes. qRT-PCR measurement of gene expression changes as a result of Zic3 knockdown. Expression was assayed in zebrafish embryos injected with either 1.7 ng and 3.4 ng or 3.4 ng alone of Zic3 MO.(XLSX)Click here for additional data file.Table S8zic3- expressing cells of the neural tube obtained by FACS at 24 hpf. Fold change was calculated by comparing expression levels in zic3-expressing cells to that in cells negative for zic3 expression.List of genes detected by microarray analysis (24 hpf). List of genes which are enriched or de-enriched in (XLS)Click here for additional data file.Table S9zic3 at 24 hpf.GO enrichment of microarray genes (24 hpf). List of GO terms enriched among genes co-expressed with (XLS)Click here for additional data file.Table S10List of microarray genes with peak (24 hpf). List of differentially expressed genes containing at least one ChIP-seq peak within 100 kb of their TSS at 24 hpf.(XLS)Click here for additional data file.Table S11zic3 and associated with at least one ChIP-seq peak at 24 hpf.GO enrichment of microarray genes with ChIP-seq peaks (24 hpf). GO terms enriched among genes co-expressed with (XLS)Click here for additional data file.Table S12in vivo enhancer assay with their genomic locations, primer sequences, and flanking genes.List of tested enhancers. List of selected enhancers tested using (XLS)Click here for additional data file.Table S13List of ChIP-seq peaks (8 hpf). List of genome-wide Zic3 binding sites identified by ChIP-seq at 8 hpf.(XLS)Click here for additional data file.Table S14List of ChIP-seq peaks (24 hpf). List of genome-wide Zic3 binding sites identified by ChIP-seq at 24 hpf.(XLS)Click here for additional data file.Text S1Supplementary experimental methods. Description of additional methods used in the study.(DOC)Click here for additional data file."} +{"text": "Tcf21 promoter and activate gene expression. Mutagenesisof SRY/SOX9 response elements in the Tcf21 promoter eliminatedthe actions of SRY. SRY was found to directly associate with theTcf21 promoter SRY/SOX9 response elements invivo during fetal rat testis development. TCF21 was found topromote an in vitro sex reversal of embryonic ovarian cells toinduce precursor Sertoli cell differentiation. TCF21 and SRY had similar effectson the in vitro sex reversal gonadal cell transcriptomes.Therefore, SRY acts directly on the Tcf21 promoter to in partinitiate a cascade of events associated with Sertoli cell differentiation andembryonic testis development.The cascade of molecular events involved in mammalian sex determination has beenshown to involve the SRY gene, but specific downstream events have eludedresearchers for decades. The current study identifies one of the first directdownstream targets of the male sex determining factor SRY as thebasic-helix-loop-helix (bHLH) transcription factor TCF21. SRY was found to bindto the Sry is located on the Y chromosome and is sufficient to inducetestis fate in the bipotential gonad Sox9 through an interaction on its testis-specific enhancerMammalian embryos have bipotential gonads which can develop into either a testis oran ovary depending on paternal and maternal chromosomal contributions. Paternaltransmission of a Y chromosome triggers testicular differentiation, whereas thecontribution of paternal X chromosome results in ovarian differentiation. Thebipotential gonad contains a pool of undifferentiated somatic cell precursors thatduring development differentiate into either testicular or ovarian (theca and granulosa) somatic cells in response toextrinsic and intrinsic factors. The gene for the transcription factorTcf21 was found to be highly expressed inthe gonads coinciding with the expression of SRY and SOX9 Tcf21 named as bHLHa23 and putit in perspective to other species Tcf21 null mutants exhibitmale-to-female sex reversal with some germs cells entering meiosis Tcf21 promoters showed at least three globalSRY-binding sites within a 2 kb promoter region. Therefore we hypothesize that SRYinteracts with Tcf21 to promote somatic cell differentiation duringmale gonadal sex determination. Observations demonstrate Tcf21 is adownstream target gene for SRY action and promotes Sertoli cell development.The process of sex determination starts with differentiation of somatic cells. SinceSRY binds to HMG box sequences on the DNA, the cell fate associated genes containingHMG box binding sites in their promoter are considered potential downstream targetcandidates. Numerous cell type-restricted basic helix-loop-helix (bHLH)transcription factors have been identified and shown to control cell fatespecification, differentiation and morphogenesis during development Tcf21 expression has been detected quite early in development inthe embryonic mesoderm, showing a similar pattern to Wt1 andGata4Tcf21 transcripts were found at the embryonic day 13 (E13)stage of testis development, after which levels are maintained during this stageof development primary Sertoli cells wereco-transfected with an Sry expression plasmid together withdifferent Tcf21 promoter fragments in a luciferase reportersystem. Similar results were obtained from both E13 testis cultures or purifiedSertoli cell cultures. The longer promoter constructs had lower luciferaseactivity with anti-HA antibody. Enriched fragments were analyzedby PCR with primers flanking the binding sites of primary cell cultures derived from E13 ratovaries. The objective was to promote an in vitro sex reversalfrom an ovarian somatic cell population to a testicular somatic cell population.TCF21 over-expression induced the expression of anti M\u00fcllerian hormone(AMH) which is a specific marker of Sertoli cell differentiation in theembryonic testis, but not expressed by the embryonic ovary or ovarian culture,The above invitro sex reversal of the embryonic ovarian cell culture to atestis cell differentiated state on a genome wide level. Microarray analysis ofthe embryonic cell cultures were investigated in the absence or presence of SRY,TCF21 or TCF21 with its potential binding partner TCF12/REB-alpha. All bHLHtranscription factors dimerize and due to the high level ofTcf12 expression in the embryonic testis , which is present inSertoli cells during early embryonic development Cbln4 gene in testis differentiation is not clearly understoodyet, this gene has been suggested as one of the downstream target genes of SRY.Using a similar approach to that taken in the current study we have found SRYdirectly interacts and regulates the promoter for neurotrophin 3(Ntf3) in vivo chromatin immunoprecipitation (ChIP) methods.Using an in vivo carrier ChIP method the current study demonstratesthat SRY interacts with the bHLH family transcription factor Tcf21during male sex determination. Observations provide insights into the mechanism ofSRY regulation of Sertoli cell differentiation and the factors regulating (e.g.TCF21) cell differentiation during male gonadal sex determination.Since the discovery of SRY as a master male sex determining factor several effortshave been made to identify its downstream target genes. WhetherTcf21 was found as one of the most highlyexpressed transcription factor genes Tcf21 mRNA is localized in the gonadal ridges and itsexpression pattern was found to be similar to Wt1 andGata4Tcf21 null mutant male mice werepartially sex-reversed Tcf21 hasmultiple roles in male sex determination and testis morphogenesis.Numerous cell type-restricted basic helix-loop-helix (bHLH) transcription factorshave been identified and shown to control cell fate specification, differentiationand morphogenesis during embryonic development, while others act later indevelopment and are required in the adult cells to maintain cellulardifferentiation. Examples include MESP in heart cell differentiation, MYOD in muscledifferentiation, and Neurogenin in neuron differentiation Tcf21 promoter harbors three SRY binding sites in the \u22122Kb promoter region, it was hypothesized that SRY may bind these response elementsand regulate the activity of the core promoter. Over-expression of SRY in theprimary embryonic testis cell culture increased the Tcf21 promoteractivity. This Tcf21 promoter activity was significantly reducedwhen all three SRY binding sites were mutated. SRY seemed to be able to activate theTcf21 promoter if any two of the three binding sites wereintact. An oligonucleotide pull-down assay was used to confirm SRY directly binds tothe Tcf21 promoter SRY binding sites Tcf21 promoter invivo a ChIP assay with chromatin samples from embryonic testisundergoing sex determination and cellular differentiation was performed. The majorlimitation in running ChIP assays with embryonic gonad samples is the amount ofchromatin required. This limitation has hindered efforts made to find downstreamtarget genes of SRY. The current study adopted the ChIP protocol developed byO'Neill et al (2006) Tcf21 gene promoter. Tcf21 appears tobe one of the downstream target genes of SRY. To identify other SRY target genes anSRY ChIP is currently being subjected to ChIP-Chip whole genome promoter tilingarrays.Most of the genes that are highly expressed during the period of male sexdetermination have SRY or SOX9 binding sites in their promoters ng sites . AlthougTcf21 null mutants have several testis phenotypes Tcf21 wasover-expressed in primary embryonic ovarian cells derived from 15 tail somite stagefemale embryos (E13) to investigate the actions of TCF21. Tcf21over-expression induced Amh expression in the female cells and soprovided evidence of its role in the induction of testis somatic celldifferentiation . AMH localization was performed using 5\u00b5g/ml primary anti-MIS antibody for 18 h and 1\u22363000 diluted Alexa Fluor 488 labeled secondary antibody using the protocol above. Non-fluorescentstaining was performed using DAB staining method.Embryonic testis sections Tcf21 promoter/reporter vectors, differentsize fragments of the rat genomic DNA upstream of the coding region was amplified to M.K.S. Rat Sry expressionconstruct was prepared as described in vitro transcribed &translated using a TnT T7 quick in vitro Transcription andTranslation kit (Promega).All the primers used in plasmid preparation are listed in the 6cells/well. Cells were maintained in 5% CO2 atmosphere in Ham'sF-12 medium (HyClone) at 32\u00b0C All cell preparation and transfections were performed according to theprotocol developed in the laboratory as previously described E13 embryos (12 to 18 tail somite-stage) were collected from timed-pregnantfemales as described above 6 Sertoli cells in 1 ml Ham's F-12 medium andincubated at 32\u00b0C for 3.5 hours. Following incubation, the cells weresubjected to hyper-osmotic shock. The medium was aspirated and 1 ml of10% glycerol in Hanks' Balanced Salt Solution was added for 3 minutes. Wells were washed twice in HBSS before freshHam's F-12 with 0.01% BSA was added. Following a 72 hourincubation cells were harvested for luciferase assays. Medium was aspiratedand 100 \u00b5l of 1\u00d7 cell lysis solution (Promega) was added perwell. Plates were frozen and thawed before cell lysate was collected. Fordetection of luciferase reporter activity 20 \u00b5l of Sertoli cell lysatewas mixed with 100 \u00b5l of luciferase substrate (Promega) and luciferaseactivity detected on a Wallac Victor II 1420 instrument.Sertoli cells cultured for 48 hours were transfected by the calcium phosphatemethod coupled with hyper osmotic shock (10% glycerol) as previouslydescribed Cells between sub 8 and 12 were transfected using Lipofectamine 2000 in vitro byusing Sry expression construct and an in vitrotranscription and translation kit according to the manufacturer'sinstruction (Promega) Tcf21 promoter with or without SRY binding site weregenerated by PCR amplification with primers listed in the 2, 10 \u00b5M ZnSO4, 10 mM Tris, pH 7.5,4% glycerol, 0.1% Triton X-100, 1 mg/mL BSA, 1 \u00b5g ofpoly(dIdC)/poly(dAdT), 0.5 mM DTT and protease inhibitor cocktail mini tabs(Sigma). Binding reaction continued for an hour at room temperature. Protein-ASepharose beads were preswollen in the cChIP incubation buffer overnight. Before use,beads were incubated with anti-HA antibody or with non-immune IgG as a negativecontrol. Following gentle centrifugation (600\u00d7 g) for 5 minutes, beadswere treated with protein-oligo mixture and incubated with gentle rotationovernight at 4 degree Celsius. In this way the antibodies, while bound to thebeads, would attach to their matching proteins and capture the oligo fragmentsspecifically bound to the proteins. Beads were washed three times with washbuffer containing 50 mM, 100 mM and 150 mM NaCl. Pulled-down oligos were elutedin a buffer containing 0.5% SDS and purified in PCR purification columnsaccording to manufacturer's instruction (Promega). Purified oligo DNA wassubjected to PCR with primers designed to flank that particular sequence ofTcf21 promoter.SRY recombinant protein with HA tag was synthesized Drosophila SL2 cells were used as a carrier.Densely-grown cells (approximately 5\u00d7107 cells) werepelleted and washed three times in ice-cold PBS, 5 mM sodium butyrate andresuspended in 0.5 ml NB buffer . Frozen testis samples were thawed, mixed with SL2 cells andhomogenized to make single cell suspension. Nuclei were pelleted,resuspended in 20 ml NB buffer, 5% (vol/vol) sucrose, pelleted andresuspended again in 5 ml digestion buffer .Following micrococcal nuclease digestion for 10 minutes at 37degree, the digested samples were gently spun (800\u00d7g) for 10 minutesand supernatant set aside on ice. The pellet was resuspended in 250 ulincubation buffer and again centrifuged gently for 10 minutes.Both the supernatants were pooled and a fraction (50 ul) out of it was keptaside to use as input. The remaining supernatant was incubated with eithernon-immune IgG or anti-SRY or anti-Gata4 antibodies at 4\u00b0C overnight.After incubation with 100 \u00b5l of preswollen protein A-Sepharose beads(Sigma) for 2 h at 4\u00b0C, the bead-bound immunoprecipitates werecentrifuged gently and washed five times with wash buffer . The protein-DNAcomplexes were incubated at room temperature with elution buffer (1%SDS in TE) and centrifuged at 11500\u00d7g at 4\u00b0C for 10 minutes.Elution was repeated two times and eluted DNA was pooled.Co-immunoprecipitated DNA was purified by proteinase K digestion,phenol/chloroform extraction, and ethanol precipitation. Final concentrationof immunoprecipitated DNA varied from 100 to 300 ng per assay. Two rounds ofPCR reactions were run for 25 cycles before evaluating samples by gelelectrophoresis. Identity of the PCR-amplified fragments was verified bysequencing. Primers used to investigate the enrichment ofTcf21 promoter fragments by PCR are listed in Carrier ChIP (cChIP) assay was adopted from O'Neill et al., (2006) http://www.ncbi.nlm.nih.gov/geo, #GSE# GSE24666 and can be alsoaccessed through www.skinner.wsu.edu, andis MIAME compliant. For genes annotation, Affymetrix annotation fileRaGene1_0stv1.na30.rn4.transcript.csv was used unless otherwise specified.Generation of affected KEGG pathways used Pathway-Express, a web-based tool freely availableas part of the Onto-Tools (http://vortex.cs.wayne.edu) mRNA processing and hybridization were performed at Genomics Core Laboratory,Center for Reproductive Biology, Washington State University, Pullman, WA usingstandard Affymetrix reagents and protocol Table S1Genes differentially expressed in F-E13 with at least 2 Treatments.(PDF)Click here for additional data file.Table S2(S2A) Differential regulation of female E13 cell culture transcriptome by ratSry expression construct. (S2B) Differential regulation of female E13 cellculture transcriptome by Tcf21 expression construct. (S2C) Differentialregulation of female E13 cell culture transcriptome by Tcf12 expressionconstruct. (S2D) Differential regulation of female E13 cell culturetranscriptome by Tcf21 plus Tcf12 expression construct.(PDF)Click here for additional data file.Table S3List of primers used in the present study.(PDF)Click here for additional data file."} +{"text": "Source Data Verification (SDV) aims to assure data quality and participant safety by checking trial data against source data in site monitoring visits. SDV is resource-intensive but its value is unclear. We evaluated SDV in an ongoing phase III RCT of three prostate cancer treatments (ProtecT) with no planned SDV.Two experienced ProtecT Data Managers reviewed 20 randomly selected participants notes at 9 hospital visits across the UK (around 7% participants per site). SDV case report forms (CRFs) were completed using hospital and trial records including baseline, eligibility, treatment and annual outcome CRFs. CRFs were entered on a separate database. Staff time, accommodation and travel costs were recorded and analysed using university costing software.639 SDV CRFs were completed (mean 4/participant) from 161 sets of available records (90% of those requested) taking a mean of 51 minutes/participant. Problems encountered included the knowledge required to interpret medical records and interrogate computer systems. SDV and original data were compared for concordance, with errors categorised as critical/major/minor. Trial CRFs were subsequently modified to increase standardisation of data collection across sites. Staff time was the major resource as was \u00a33,126 of subsistence/travel costs .SDV was conducted at 9 site visits on around 5% of participants in a cancer trial. Logistical issues hindered data collection. Comparison of SDV data against trial data helped improved trial CRFs. Further analysis will evaluate data quality gains against the considerable costs."} +{"text": "We demonstrate the feasibility of 3D myocardial perfusion CMR at 3 Tesla against fractional flow reserve (FFR) in 53 patients for the detection of flow-limiting coronary artery disease and show good agreement between the techniques. This technique shows excellent diagnostic sensitivity and specificity and may offer an alternative method of detecting ischaemia for the purpose of guiding revascularisation and risk stratification.Three-dimensional (3D) myocardial perfusion cardiovascular magnetic resonance (CMR) has recently been proposed to overcome the limited spatial coverage of conventional perfusion CMR methods1. The method has shown good diagnostic accuracy for the detection of coronary artery disease determined by quantitative coronary angiography (QCA)2. However the relationship between the severity of a coronary stenosis on QCA and its functional significance is variable. Pressure wire-derived fractional flow reserve (FFR) <0.75 correlates closely with objective evidence of reversible ischemia and it has been demonstrated that ischaemia-guided PCI confers a prognostic benefit.To determine the diagnostic accuracy of whole heart 3D myocardial perfusion CMR against invasively determined FFR.Fifty-five patients referred for angiography underwent rest and adenosine stress 3D myocardial perfusion CMR at 3Tesla . Perfusion was scored visually on a patient and coronary territory basis with a score from 0-3. Ischaemic burden was calculated by quantitative segmentation of the volume of hypoenhancement. The FFR was measured in vessels with >50% severity stenosis. Fractional flow reserve <0.75 was considered hemodynamically significant.Two patients were excluded . From the remaining fifty-three patients and 159 coronary vessels, 64 underwent pressure wire assessment and 39 had an FFR<0.75 (Figure3D CMR stress perfusion can detect functionally significant coronary artery disease with excellent sensitivity, specificity and predictive values when compared with FFR.3D CMR perfusion imaging may offer an alternative method of detecting ischaemia for the purpose of guiding revascularisation and risk stratification.SP is funded by British Heart Foundation fellowship FS/10/62/28409.SP/EN receives research grant support from Philips Healthcare."} +{"text": "Phodopus sungorus papillomavirus type 1 (PsPV1), isolated from an anogenital lesion of a Siberian hamster. PsPV1 is taxonomically classified in the genus Pipapillomavirus and is most closely related to Mesocricetus auratus papillomavirus 1 (MaPV1).We report the complete genomic sequence of Pipapillomavirus currently consists of six PV types: MaPV1, MmuPV1, AsPV1, McPV2, MmiPV1, and RnPV1, isolated from a wide variety of rodent animal species, including the Syrian golden hamster (Mesocricetus auratus), house mouse (Mus musculus), wood mouse (Apodemus sylvaticus), Southern multimammate mouse (Mastomys coucha), Eurasian harvest mouse (Micromys minutus), and Norway rat (Rattus norvegicus), respectively , which was isolated from an anogenital lesion of a Siberian hamster (Phodopus sungorus). No PVs have been previously shown to infect this particular animal species.Papillomaviruses (PVs) are a large and diverse group of small double-stranded DNA viruses that are etiologically linked with various benign and malignant lesions of the skin and mucosa. They infect humans and at least 54 other animal species . To datehttp://blast.ncbi.nlm.nih.gov/Blast.cgi) of the partial L1 sequence revealed that PsPV1 clusters to the genus Pipapillomavirus. A complete viral genome was amplified using inverse long-range PCR with PsPV1 L1 gene-specific primers and cloned using a TOPO XL PCR cloning kit . The complete genomic sequence was determined by the primer walking strategy at Microsynth AG and assembled and characterized using Vector NTI Advance 11 software (Invitrogen). Phylogenetic analysis was performed using a maximum likelihood algorithm , which was originally obtained from an oral lesion of Syrian hamster (M. auratus) and two late open reading frames (ORFs) (L1 and L2), but no E4 and E5 ORFs. The noncoding region of 416\u00a0bp is typically positioned between the L1 and E6 ORFs. Phylogenetic analysis confirmed that PsPV1 is a member of the Pipapillomavirus genus and suggests a possible etiological role of PsPV1 in the development of anogenital neoplasms in Siberian hamsters, which needs to be investigated further.In conclusion, the present study expands the heterogeneity of the HG939559.The complete genome sequence of PsPV1 has been deposited at DDBJ/ENA/GenBank under the accession no."} +{"text": "The Croatian Cardiovascular Tissue Bank (CTB) was established in June 2011 at University Hospital Centre Zagreb (UHC). Cardiovascular tissues processed in CTB were retrieved and processed according to validated protocols and in compliance with laws and regulations applied in tissue establishments.Cardiovascular tissue allografts (CVA) were retrieved from recipients of heart transplants (RHT) and multiorgan donors (MOD) which fulfill donor selection criteria. Processing of CVA was performed in CTB clean room (GMP Class B). Initial processing included surgical dissection, measurements, morphology and functional assessment and collection of quality control samples. CVA were incubated in antibiotic containing medium for 24-48 hours. Then CVA were cryopreserved in solution containing 10% of dimethylsulphoxyde and stored in vapor phase of liquid nitrogen. Cryopreserved tissues remained in the quarantine until all quality control results were collected.During time period from June 2011 to March 2013, 63 cardiovascular tissue allografts were retrieved from 35 donors (17 RHT and 18 MOD). 32 CVA were accepted for clinical use. Accepted tissues were as follows 15 aortic valves, 13 pulmonary valves, 4 thoracic aortas. 10 CVA remained in quarantine and 21 CVA were discarded. Reasons for CVA to be discarded were microbiological, serological and morphological. 12 CVA were successfully transplanted since June 2012. Indications were infectious (7 patients), non-infectious (2 patients) and congenital (1 patient).The cardiovascular tissue bank was successfully established at UHC Zagreb. CTB needs to work on the rising of public and surgeons awareness about cardiovascular tissue donation in order to achieve large selection of high quality cardiovascular tissue allografts. Initial clinical results are encouraging."} +{"text": "Many pharmacologically-relevant polymorphisms show variability among different populations. Though limited, data from Caucasian subjects have reported several single nucleotide polymorphism (SNPs) in folate biosynthetic pathway. These SNPs may be subjected to racial and ethnic differences. We carried out a study to determine the allelic frequencies of these SNPs in an Indian ethnic population.Whole blood samples were withdrawn from 144 unrelated healthy subjects from west India. DNA was extracted and genotyping was performed using PCR-RFLP and Real-time Taqman allelic discrimination for 12 polymorphisms in 9 genes of folate-methotrexate (MTX) metabolism.MTHFR 677T (10%) and 1298 C (30%), TS 3UTR 0bp (46%), MDR1 3435T and 1236T (62%), RFC1 80A (57%), GGH 401T (61%), MS 2756G (34%), ATIC 347G (52%) and SHMT1 1420T (80%) in healthy subjects (frequency of underlined SNPs were different from published study data of European and African populations).Allele frequencies were obtained for The current study describes the distribution of folate biosynthetic pathway SNPs in healthy Indians and validates the previous finding of differences due to race and ethnicity. Our results pave way to study the pharmacogenomics of MTX in the Indian population. It is an essential cofactor for Recently, several studies have described SNPs of the genes involved in folate metabolism6The data on the distribution of SNPs (folate pathway) in the Indian population are sparse. Any study on the frequencies of SNPs in disease must be preceded by their distribution in the healthy population. The present study aims to determine the allelic frequency of SNPs across intracellular folate metabolic pathway in healthy Indian subjects. Allele frequencies were also compared with previously reported frequencies by others to address the racial and inter-ethnic differences.One hundred and forty four unrelated healthy subjects of either sex were enrolled in the study from the outpatient rheumatology referral service of Centre for Rheumatic diseases, Pune (west India) from April 2007- January 2008. There were 70 males and 74 females with mean age (SD) of 23.22 (5.3) years. The study protocol was approved by the Ethics Committee of Centre for Rheumatic Diseases, Pune. Subjects willing to participate provided consent as per the guidelines from the Institutional ethics Committee.MTHFR: Methylenetetrahydrofoate reductase; TS: Thymidylate synthase; RFC1: Reduce folate carrier1; MS: Methionine synthase; SHMT1: Serine hydroxymethyltransferase1; MDR1: Multidrug resistant protein1; GGH: \u03b3 glutamyl hydrolase; ATIC: Aminoimidazol carboxamide ribonucleotide transformylase; MTRR: Methionine synthase reductase. Genotyping was performed using PCR-RFLP technique for MTHFR A1298C (rs1801131) and C677T (rs1801133), TS 5\u2019UTR repeat and 3\u2019UTR deletion, RFC1G80A (rs1051266), MS A2756G (rs1805087), MDR1C3435T (rs1045642) and C1236T (rs1128503), GGH C401T (rs3758149), MTRR A66G (rs1801394) polymorphisms ATIC C347G (rs2372536), SHMT1C1420T (rs17829445) polymorphismsTS which were directly visualized after the PCR. Real-time Taqman allelic discrimination assays were performed according to protocols provided by the manufacturer . Samples containing mutants were reanalyzed to ensure the accuracy of the method. There was 100 per cent reproducibility.Post-consent, peripheral blood sample (4-5 ml) was drawn from each healthy subject, and genomic DNA was extracted using Miller\u2019s protocolA1298C rs01131 andP< 0.05 was considered statistically significant. The observed genotype frequencies of polymorphisms studied were compared with expected frequencies according to Hardy-Weinberg equilibrium (HWE) using \u03c72 tests.Statistical analysis was performed using the Graph Pad Prism statistical software (San Diego CA. USA). Allele frequencies were determined for 12 polymorphisms in nine genes in the folate-MTX metabolic pathway in 144 healthy subjects. The frequency of each allele in the study population is given in the MTHFR 677T (10%) and 1298 C (30%), TS 3UTR 0bp (46%), MDR1 3435T and 1236T (62%), RFC1 80A (57%), GGH 401T (61%), MS 2756G (34%), ATIC 347G (52%) and SHMT1 1420T (80%). The complete genotype distribution for healthy subjects is represented in We examined allele frequencies for 12 polymorphisms in folate and MTX metabolism among healthy subjects and compared them with the allele distribution in other ethnic groups . Allele MTHFR 677T variant allele frequency in European population was higher than Indian healthy subjects (10%) while TS 3 UTR 0bp (deletion) polymorphism was lower in European than Indian (46%). There was no difference in distribution of MTHFR 1298C, MDR1 1236T and MDR1 3435T variant allele frequencies between Indian and European healthy subjects. The occurrence of MTHFR 677T , MTHFR 1298C , MDR1 3435T and MDR1 1236T variant alleles was significantly lower in Africans as against Indian healthy subjects.Healthy subjects from our study were compared with healthy subjects from European, African and Indian population . MTHFR 6GGH 401T, RFC1 80A and MDR1 1236T variant alleles. The occurrence of GGH 401T (61%) andRFC1 80A (57%) in our healthy subjects was higher than north Indian subjects 25 and 28 per cent (P<0.0001) respectively while MDR1 1236T was higher in north Indians (72%) than our healthy subjects . Thus the current report supports the previous findings that the allele or haplotype frequencies of several important polymorphisms in folate pathway vary with raceComparison of our healthy subjects with Indian study from north India reveals that there was significant difference in the occurrence of MTHFR 1298C and 677T, TYMS 3\u2019 UTR deletion and MDR1 3435T and 1236T in healthy subjects are different in Indian subjects as compared to Europeans and AfricansGGH 401T, RFC1 80A and MDR1 1236T variant alleles within Indian population. This intra-ethnic difference can be because Indian population is a conglomeration of multiple culture and evolutionary histories. The evolutionary antiquity of Indian ethnic groups and subsequent migration from central Asia, west Asia and southern China has resulted in a rich tapestry of socio-cultural, linguistic and biological diversityThe allele frequencies of per se to impair folate-mediated one-carbon metabolic pathways and contribute to increased risk of several disorders of folate deficiency6SNPs have been reported folypolyglutamate synthase (FPGS) and dihydrofolate reductase (DHFR). We report ethnic differences in the SNPs in genes coding folate biosynthetic metabolic intracellular pathway. It may not be appropriate to extrapolate the findings of genetic associations influencing folate antagonist treatment response in subjects belonging to Caucasian and African ethnicity to the Indian population. Thus knowledge of allelic frequency distribution within a population can be useful in optimizing doses for therapeutic efficacy, identifying potential risk groups for adverse drug reactions and explaining therapeutic failures.To our knowledge this is the first report on 12 polymorphisms in 9 genes of folate metabolic pathway in Indian population. We have not analyzed polymorphisms in"} +{"text": "There are interrelations among coronary stiffness, wall thickening and remodeling pattern in older hypertensive patients. MR techniques are able to provide both the morphology and the biomechanic information of the coronary wall in a single scan.Coronary artery remodeling is an indicator of coronary artery disease. Morphological and biomechanical changes accompany with coronary wall remodeling. Such changes can be noninvasively detected using MR imaging and may be used to predict cardiovascular events. The aim of this study is to assess interrelations among various imaging markers of the coronary remodeling in older hypertensive patients.This study was approved by the institutional review board. Two-dimensional black-blood coronary wall MR imaging and three-dimensional whole-heart coronary MR angiography (imaging data acquired in both systole and in diastole) were performed on 65 asymptomatic hypertensive patients (73.4 years \u00b1 5.5). Coronary vessel area, wall area, lumen area, wall thickness were measured. The percent of the coronary wall occupying the vessel area (PWOV) and coronary distensibility index (CDI) (defined as [(Lumen systolic - Lumen diastolic) / (Lumen diastolic x Pulse pressure)] x 1000) were calculated . Transverse coronary segments were assigned to two groups using mean PWOV as an ad hoc cutoff point. Coronary indices were compared between the two groups.Totally 259 coronary segments were eligible for analysis (mean PWOV 74.5%). The CDI (5.30 \u00b1 2.60 mmHg-1) was significantly correlated with mean wall thickness, max wall thickness . The PWOV was significantly correlated with mean wall thickness (r = 0.647), max wall thickness (r = 0.603) and lumen area . One hundred and forty segments (group 1) had PWOVs higher than 74.5%, while 119 segments (group 2) had PWOV lower than 74.5%. Segments in group2 had a significantly lower mean and max wall thickness , a larger vessel area , a larger lumen area and a higher CDI compared with those of segments in group 1. For the 10 randomly chosen cases, good intra-observer and inter-observer agreement was found on 43 coronary segments. The scan-rescan test showed low variation between coronary measurements in 38 coronary segments from the 10 randomly chosen cases .A set of images for coronary segments with different PWOV from a single patient is shown in Figure Aterial stiffness is correlated with wall thickness of the remodeled coronary artery in older hypertensive patients. Reflecting relaions between wall thickening and lumen area variation, PWOV has the potential to become a quantitative index of coronary remodeling patterns (such as positive or negative remodeling) in imaging studies.National Institute of Health (R01HL089695)American Heart Association (10CRP3050051)"} +{"text": "Chronic ankle instability can affect activities of daily living in patients. While many studies have indicated postural control deficits in these patients ,2, the ePostural stability in CAI patients (n=8) and healthy subjects (n=10) was measured using the Force Plate. Eight positions concluded two different stances (double & single) with closed or opened eyes. All positions concurrently were done with a cognitive task. Anterior/posterior (Rfa) and medial/lateral (Rsw) mean sway, area and mean velocity quantified static postural control .Mean sway significantly increased in patients in the anterior/posterior (single and double leg stance) and medial/lateral (single leg stance) directions (P<0.05). While performing a dual task anterior/posterior mean sway decreases within the patients group on the impaired leg stance (P<0.05). Area significantly increased in patients in single leg stance but decreased in the bilateral standing positions except open eyes with cognitive task. Mean velocity significantly decreased (single leg stance) and increased (double leg stance). No difference is seen in the healthy subjects.Postural control deficits were identified in participants with chronic ankle instability. In view of the fact that a cognitive task resulted in decreasing displacement of centre of pressure in patients, this method may identify as an examination and a plan of treatment for affecting on ankle stabilizing factors."} +{"text": "Continuous glucose monitoring (CGM) in ICUs has the potential to improve patient safety outcomes. The GluCath Intravascular CGM System uses a novel quenched chemical fluorescence sensing mechanism to measure glucose concentration in venous or arterial blood (BG). This is the first report of this system deployed for 48 hours in both arteries and veins of ICU patients.In vivo calibration was performed at 1 and 2 hours, then each morning. Glucose values were recorded every 10 seconds by the system. Hourly arterial reference samples were analyzed via Radiometer ABL 800 Flex Blood Gas Analyzer (BGA).This ongoing clinical study evaluates up to two sensors per subject in 20 patients undergoing cardiac surgery. An arterial sensor is deployed via a standard 20 G radial artery catheter inserted for routine care and an optional venous sensor is deployed percutaneously in an upper arm vein. Data are presented from the first five patients. Outcome measures are qualitative and quantitative . Sensors were inserted shortly after ICU admission, with ultrasound guidance for venous sensor insertion. n = 2) was 9.3% CV. Arterial sensor accuracy compared with BGA was 5.5% MARD. One hundred percent (202/202) of arterial sensor measurements met ISO 15197 criteria (within =/-20% of reference measurements if BG = 4.2 mmol/l; Figure Arterial sensors were successfully deployed in all five patients and did not interfere with clinical care, blood pressure monitoring or sampling. One arterial catheter failed resulting in sensor removal at 36 hours. The venous sensor was deployed in three patients, but removed from two patients due to thrombosis identified during surveillance ultrasound examinations. A total of 202 reference BG samples ranging from 5.3 to 11.3 mmol/l were collected. Precision between arterial and venous sensors (The GluCath System measured glucose concentration continuously in cardiac surgery ICU patients. Arterial catheter deployment did not appear to compromise line function or patient care. Percutaneous venous deployment was feasible, but may be associated with risk of local venous thrombosis."} +{"text": "The purpose of this study was to investigate the relationship between the extent of hyperenhancement and left ventricle maximal wall thickness (mWT) detected by LGE CMR and clinical events of nonsustained ventricular tachycardia (NSVT), implantation of cardioverter defibrillator (ICD) or diastolic heart failure in HCM patients that underwent LGE CMR.Late gadolinium enhancement cardiac magnetic resonance (LGE CMR) imaging has been used to detect myocardial hypertrophy and scar/fibrosis in Hypertrophic cardiomyopathy (HCM) patients. The presence of hyperenhancement has been associated with progressive ventricular dilation, ventricular arrhythmias and clinical risk factors for sudden cardiac death.Under IRB approved protocol a total of 82 HCM patients underwent LGE CMR images using a 2D PSIR TurboFLASH protocol after administration of 0.2 mmol of gadopentetate dimeglumine per kilogram of body weight. The presence of LGE was assessed using automated software: percentage of scar and maximal wall thickness were calculated. Percentage of scar was compared between patients with mWT <2.5 cm and \u22652.5cm The mean values of percentage scar and mean maximal wall thickness between patients with/ without clinical events were compared.Scar was detected in 74.5 % patients and clinical events were present in 54 (%) patients. Only one patient without scar had NSVT and had ICD implanted. There was a significant difference between mWT in patients with scar 2.0 cm) and no scar 1.5 cm) p =0.002. Scar % and mean mWT were significant higher in patients with positive clinical events(Table .5 cm p =.0 cm andIn conclusion, HCM patients with a mWT above 2.5 cm maybe at increased risk of events regardless of the amount of myocardial scar.None."} +{"text": "Progressive hemifacial atrophy (PFH) and linear scleroderma en coup de sabre (LSCS) may be accompanied by neurologic symptoms and other extra-cutaneous manifestations.To investigate and compare clinical/immune characteristics of patients with LSCS and PFH.Retrospective study of 12 patients presenting at 2 pediatric dermatology clinics with linear scleroderma and/or hemifacial atrophy9 patients presented with LSCS, followed in 5 of them by progressive hemifacial atrophy (PFH) within 2 years. 3 patients presented with PFH, all 3 had additional scleroderma lesions. Their median(range) age at presentation was 8(4-17)yrs.Extracutaneous manifestations were equally found in LSCS and PFH+LSCS patients. They comprised asymmetry of tooth arches/missing teeth(1), ophthalmologic problems (4), epileptic seizures with hyperintense signals on MRI and hypoperfusion on SPECT in 1 LSCS (fig) and 1 PFH patient, severe migraine attacks(2), polyarthritis(2), vitiligo(1), celiac disease(1).ANA were found in 3 LSCS, 2 PFH+LSCS patients. Distinct oligoclonal IgG bands were found in CSF (not in blood) in patients with PFH and seizures. A skin biopsy in a PFH lesion showed fibrosis associated with a lymphocytic infiltrate, IgM and IgG deposits. A brain biopsy in one PFH patient with epilepsy was consistent with Rasmussen encephalitis.In patients with severe skin +- neurological lesions, treatment with steroids/high-dose MTX resulted in improvement/stabilization of clinical and MRI abnormalitiesOur case series endorses the concept of a single disease spectrum encompassing LSCS and PFH, with a common immune-inflammatory pathogenesis. A possible relationship with RE is suggested as well."} +{"text": "Hereditary non-polyposis colorectal cancer or Lynch syndrome (LS) is characterized by mismatch repair (MMR) loss of function. The aim of this study was to compare the clinical characteristics and family histories of MMR mutation-positive and mutation-negative patients with tumor studies suggestive for LS.Patients with loss of MLH1/MSH2 on immunohistochemistry (IHC) or with microsatellite instability (MSI)-high tumors were identified in our institutional LS database (February 1992\u2013June 2010). Patients who subsequently underwent MLH1/MSH2 mutation analysis were reviewed. Patients with no identifiable MLH1 germline mutation were excluded if MLH1 promoter methylation was present or not assessed. Patients with variants of uncertain significance were also excluded. Demographics, clinical characteristics, mutational testing, and family histories of patients were analyzed.Of 92 patients with informative tumor studies who underwent germline mutation testing with unequivocal results, 61 (58.7%) were mutation-positive and 31 (29.8%) were mutation-negative. The mutation detection rate for MLH1 and MSH2 was 51.2% and 78.4%, respectively. A significant difference (p=0.006) in the proportion of MLH1 and MSH2 defects was found between mutation groups was diagnosed in 76 patients (82.6%; Table In this population of patients with presumptive LS diagnoses based upon tumor studies, mutation-negative patients had significant personal cancer histories concerning for LS with slightly later ages of initial LS cancer and CRC diagnosis. However, their family histories were significantly less suggestive as nearly one-third of patients were missed by Bethesda criteria and over 85% by Amsterdam I/II criteria. This suggests that both established clinical criteria and germline mutation analysis may fail to detect a significant number of patients with presumed LS and supports the use of routine MSI/IHC analyses to identify these otherwise undetectable high-risk families."} +{"text": "Quantitative cardiovascular magnetic resonance (CMR) myocardial perfusion imaging has the potential to evolve into a routine clinical method allowing for the assessment of myocardial blood flow (MBF). Multiple quantification pathways are available based on different algorithms. These algorithms involve complex modeling and quantitative results may not necessarily be the same. At present it remains unclear which algorithm is the most accurate. An isolated perfused, magnetic resonance (MR) compatible pig heart model allows very accurate titration of MBF and in combination with high-resolution assessment of fluorescently-labeled microspheres represents a near optimal platform for validation. We sought to investigate which algorithm is most suited to quantify myocardial perfusion by CMR imaging at 1.5 and 3 Tesla using state of the art CMR perfusion techniques and quantification algorithms.First-pass CMR perfusion was performed in a MR compatible blood perfused pig heart model. We acquired perfusion images at resting flow (100%), 50% flow and during adenosine induced hyperemia in control and coronary occlusion conditions. MR myocardial perfusion imaging was performed at 1.5 Tesla (n=4) and at 3 Tesla (n=4). Fluorescently-labeled microspheres and externally controlled coronary blood flow served as reference standards for comparison of different quantification strategies, namely Fermi function constrained deconvolution, autoregressive moving average modeling, deconvolution using an exponential basis and deconvolution using a B-spline basis.All CMR derived MBF estimates agreed well with microsphere results. The best correlation was achieved with Fermi function constrained deconvolution both at 1.5 Tesla and at 3 Tesla . Fermi deconvolution correlated significantly better with the microspheres than all other methods at 3 Tesla . Whilst it was superior to B-spline at 1.5 Tesla (p=0.001) it was not statistically superior to exponential deconvolution and ARMA deconvolution at 1.5 Tesla (p>0.05).The weakest correlation at 1.5 Tesla was found using B-spline deconvolution and at 3 Tesla using exponential deconvolution .CMR derived quantitative blood flow estimates correlate with true myocardial blood flow in a controlled animal model. Amongst the different techniques, Fermi function constrained deconvolution was the most accurate at both field strengths. Quantitative CMR perfusion based on Fermi function deconvolution may therefore emerge as a useful clinical tool providing accurate blood flow assessment.Andreas Schuster is a British Heart Foundation (BHF) Clinical Research Fellow (FS/10/029/28253) and received grant support from the BHF (RE/08/003) and the Biomedical Research Centre (BRC-CTF 196). Matthew Sinclair receives support from the Engineering and Physical Sciences Research Council (EP/H046410/1). Jeroen P. H. M. van den Wijngaard is funded by a VENI grant of the Netherlands Organization for Scientific Research (NWO/ZonMw 916.11.171). This study was further supported by grants to the AMC from the Netherlands Heart Foundation (NHS 2006B186 and 2006B226), the Netherlands Organization for Health Research and Development (ZonMw 91105008 and 91112030), and the European Community (FP7-2007-224495: euHeart project).Nicolas Smith receives grant support from Wellcome Trust and Engineering and Physical Sciences Research Council . Amedeo Chiribiri was funded by the Wellcome Trust and EPSRC under grant number WT 088641/Z/09/Z. Eike Nagel receives grant support from BHF (RE/08/003), the Wellcome Trust and Engineering and Physical Sciences Research Council and the National Institute for Health Research (NIHR) via the comprehensive BRC award to Guy's and St Thomas' NHS Foundation Trust in partnership with King's College London and King's College Hospital NHS Foundation Trust."} +{"text": "A points-based scoring system defines levels of familial risk for colorectal neoplasia. More than 7 points defines a high-risk group that should undergo colonoscopy every 3 years. Family History Scoring was applied to families fulfilling Amsterdam I criteria and familial colorectal cancer type X to compare colon cancer risk.Amsterdam I (AmI) and familial type X (famX) families identified from Jagelman Registry database were scored. Affected probands (AP), unaffected probands (UP), affected sibling (AS), and unaffected sibling (US) of the proband, and a child of each sibling were scored. Median, range, and standard deviation were compared for each syndrome to determine level of risk.200 patients were included. 181 matched AmI criteria with genetic testing, 19 matched criteria for famX. 37 families with an AP and 12 with an UP matched AmI criteria. 4 with an AP and 3 with an UP that matched criteria for famX. Level of risk for the two syndromes were compared (Table Family history scoring system has shown that families fulfilling Amsterdam I criteria and familial type X share similar familial risk level for developing colon cancer. Both show dominant inheritance, but molecular pathways differ."} +{"text": "The study of cholesterol homeostasis is largely restricted to the use of animal models and immortalized cell lines that do not recapitulate those key aspects of normal human hepatocyte function that result from genetic variation of individuals within a population. Hepatocyte-like cells (HLCs) derived from human embryonic and induced pluripotent stem cells can provide a cell culture model for the study of cholesterol homeostasis, dyslipidemias, the action of statins and other pharmaceuticals important for cardiovascular health. We have analyzed expression of core components for cholesterol homeostasis in untreated human iPS cells and in response to pravastatin. Here we show the production of differentiated cells resembling periportal hepatocytes from human pluripotent stem cells. These cells express a broad range of apolipoproteins required for secretion and elimination of serum cholesterol, actively secrete cholesterol into the medium, and respond functionally to statin treatment by reduced cholesterol secretion. Our research shows that HLCs derived from human pluripotent cells provide a robust cell culture system for the investigation of the hepatic contribution to human cholesterol homeostasis at both cellular and molecular levels. Importantly, it permits for the first time to also functionally assess the impact of genetic polymorphisms on cholesterol homeostasis. Finally, the system will also be useful for mechanistic studies of heritable dyslipidemias, drug discovery, and investigation of modes of action of cholesterol-modulatory drugs.Hepatocytes play a central and crucial role in cholesterol and lipid homeostasis, and their proper function is of key importance for cardiovascular health. In particular, hepatocytes endogenously synthesize large amounts of cholesterol and secrete it into circulating blood via apolipoprotein particles. Cholesterol-secreting hepatocytes are also the clinically-relevant cells targeted by statin treatment Chronic exposure to high serum cholesterol is the single largest risk factor for the development of atherosclerosis and results from perturbed cholesterol homeostasis Apolipoproteins (APOs), a family of more than 20 proteins, are key components of lipoprotein particles Within the liver, functionally-specialized hepatocytes are sequentially organized into periportal, mid-zonal, and perivenular zones along the sinusoidal tubules of the liver cell plate within liver lobules Current model systems to study hepatocyte function include the use of primary human hepatocytes, animal models, and transformed hepatic cell lines. These models suffer from substantial and fundamental deficiencies. Primary hepatocytes are unstable in cell culture, and have variable and short-lived CYP activity In the current study we have reprogrammed skin-derived dermal fibroblasts from healthy human donors 2 matrigel (BD Sciences Inc.) in mTESR1 (Stemcell Technologies Inc.). Pluripotent cells were maintained under above conditions throughout the experiment. Embryoid bodies were generated by transfer of mechanically dissociated stem cell colonies into culture plates coated with a 2% poly-2-hydroxyethyl-methacrylate (poly-hema) solution and cultured for 20 days in DMEM/F12 supplemented with 20% knock out serum replacement, 1x NEAA, 2mM glutamine and 100 \u03bcM \u03b2-mercaptoethanol. Hepatocytes were maintained on 35 \u03bcg/cm2 matrigel (BD Sciences Inc.) in hepatocyte growth medium supplemented with 10 ng/ml oncostatin M (Prospec Inc.) and dexamethasone (Sigma Inc.).Primary human dermal fibroblasts were grown according to the vendor's instruction in MEM supplemented with 15% fetal bovine serum, 1x non-essential amino acids and 2mM glutamine (Invitrogen Inc.). Cultures were passaged at 80-90% confluency. WA09 (H9) cells were obtained from WICELL at passage 19, adapted to growth on 35 \u03bcg/cm5 fibroblasts either with a mixture of lentivirus particles (Openbiosystems Inc.) containing genomes encoding OCT4, SOX2, KLF4 and human c-MYC at a multiplicity of infection (moi) of 5\u22365\u22365\u22362.5 (hDF6) or, for passage 13 cells (hDF1), with a similar mixture of lentivirus particles expressing the above genes (Addgene) and a LMNA shRNA at a moi of 5 in a doxycycline dependent fashion to create an ESC-like chromatin-state \u22125 (data not shown).Human dermal fibroblasts from 3 year and 43 year old healthy unaffected females were reprogrammed using a published protocol Transduced fibroblasts were maintained in DMEM/10% FCS until day 4 post transduction. On day 3 post transduction, fibroblasts were passaged onto feeder cells and on day 4 the medium was changed to embryonic stem cell culture medium consisting of DMEM/F12 (Invitrogen) containing 20% knock out serum replacement (Invitrogen), 1x non-essential amino acids, 1mM glutamine, 100 \u03bcM \u03b2-mercaptoethanol and 5 ng/ml FGF2. Media were changed daily and in the case of doxycycline-inducible expression of reprogramming factors, doxycycline was administered daily at a concentration of 2 \u03bcg/ml until day 18 post transduction. Candidate iPS cells were identified by colony morphology and transferred into fresh culture wells after 25 (hDF1) and 30 days (hDF6), respectively and maintained under feeder free conditions on matrigel in the presence of mTESR1. Viral titers were obtained by transduction at defined dilutions of the individual lentivirus into sentinel fibroblasts through immunofluorescent detection of the respective reprogramming factor.6 cells/ml. For the compensation between FITC and PE, anti-mouse IgG BD CompBeads Plus (BD Biosciences Inc.) were used. About 2\u00d7104 events (cells) were sorted using a FACS Calibur flow cytometer. FACS profiles were analyzed using CellQuest Pro Software (BD Biosciences Inc.) and FlowJo software v9 (Tree Star Inc.).Cells were harvested at 70% of confluence and a single cell suspension was prepared by dissociation with Accutase (Invitrogen) for 5 min at 37\u00b0C and filtering through a 40\u03bcm cell strainer (BD Biosciences). Cells were then fixed with 4% paraformaldehyde for 15 min at room temperature, washed once with PBS and resuspended in a staining buffer (0.2%BSA in PBS). After 30 min on ice cells were incubated with anti-SSEA4 antibody conjugated to phycoerythericin and anti-Tra1-60 antibody conjugated to fluorescein (BD Biosciences Inc.). The cells are then washed two times in PBS and then resuspended in staining buffer at a concentration of 10ES/iPS cells were grown in mTESR1 on matrigel until colonies of pluripotent cells had a diameter of approximately 1\u20132 mm before processing for embryoid body (EB) differentiation. Colonies were cut into smaller fragments and then dislodged mechanically. An average of 100 cell clumps were then transferred into ultralow adhesion plates (Becton Dickinson) and cultured for 21 days in DMEM/F12 supplemented with 20% knock-out serum replacement (Invitrogen Inc.), non-essential aminoacids (Invitrogen Inc.) and 2 mM glutamax (Invitrogen Inc.), 0.5 ng/ml FGF2 and 0.1 mM \u03b2-mercaptoethanol. Embryoid bodies (EBs) were then harvested directly into RLT (Qiagen Inc.) for preparation of total RNA.5 cells/cm2. The cells were grown to approximately 50% confluency in mTeSR1 media (StemCell Technologies) and then differentiated into the three germ layers by replacing mTeSR1 media with lineage specific media (R&D Systems). After 4 days of culture, the cells were analyzed for Sox17 (endoderm), Otx2 (ectoderm) or Brachyury (mesoderm) protein expression by immunofluorescence.Pluripotent stem cells were differentiated into derivatives of ecto-, meso-, and endodermal lineage cells using the Human Pluripotent Stem Cell Functional Identification kit according to the manufacturer's instructions (R&D Systems). Briefly, cells were harvested as a single-cell suspension and plated onto matrigel at a density of 10ES/iPS cells were grown in mTESR1 on matrigel until colonies of pluripotent cells had a diameter of approximately 1\u20132 mmbefore differentiation was initiated. Differentiation was performed by methods similar to those previously described Stage 3B HLCs were cultured for 48 hours in HCM containing 10 \u03bcM pravastatin (Sigma Inc.). Culture medium was retained, cleared of cellular debris by centrifugation at 14,300\u00d7 g and an aliquot was assayed for soluble cholesterol content using the Amplex Red Cholesterol assay kit according to the manufacturer's instructions (Invitrogen Inc.). Cholesterol amounts were normalized to total DNA content of adherent cells.RNA was prepared in triplicates from biologically independent samples. Cells were lysed in RLT buffer (Qiagen Inc.) containing 1% \u03b2-mercaptoethanol and processed using RNeasy ion exchange column chromatography according to the manufacturer's instructions (Qiagen Inc.). Purified RNA was quantified by UV spectroscopy.DNA was prepared in triplicates from biologically independent samples. Cell pellets were resuspended in digestion buffer supplemented with 60 \u03bcg/ml DNase-free RNase and incubated at RT for 10 min. DNA was released overnight at 56\u00b0C in the presence of 0.5% SDS and Proteinase K (100\u03bcg/ml) and peptides were removed by phenol extraction using phase lock gels according to the manufacturer's instructions (Qiagen Inc.). DNA was quantified by UV spectroscopy.TM cDNA synthesis kit (BioRad) was used for cDNA synthesis. Quantitative real-time PCR analyses were performed using the iTaq Fast SYBR Green Supermix with ROX (BioRad) and a 7500 Fast Real Time PCR System (Applied Biosystems). Data were analysed by means of 2\u2212\u0394Ct method for relative quantification An amount of 500 ng of total RNA and the iScriptAntibodies used in this study included polyclonal anti-human albumin antibodies , monoclonal anti-human-AFP, -APOA1, -APOA2, -APOC3 and -LDLR antibodies , a monoclonal anti-human SOX17 antibody , a biotinylated human TRA1-60 antibody and a polyclonal anti-human NANOG antibody (Millipore). For immunofluorescent detection of the antigens, primary antibodies were diluted 1:200 and Alexa 488 and Alexa 594 coupled secondary antibodies (Invitrogen) 1:1000. Cells were fixed in 4% paraformaldehyde, washed three times in TBS containing 1% bovine serum albumin (BSA), permeabilized in 0.1% Triton X-100, blocked in TBS containing 3% BSA and incubated with the appropriate primary antibodies in TBS containing 3% BSA for a minimum of two hours. After three washes cells were incubated with the appropriate secondary antibodies in TBS containing 3% BSA for a minimum of two hours, washed three times and imaged. Nuclei were marked using TBS containing 1% BSA and 4ng/ml of 4\u2032, 6\u2032-diamidino-2-phenylindole (Invitrogen Inc.). For quantitative immunofluorescence 2000 fixed cells were spun onto poly-lysine coated coverslips and processed as described above. A minimum of 200 cells per set was scored for antigen positivity and negativity and expressed proportionally for graphical representation. Proportions were determined by dividing the number of cells counted with a given pattern by the total number of cells (n) and all samples were analysed in triplicates from biologically independent samples. The standard deviation was determined using the equation s \u00d7 sqrt(p(1-p)/n), where p is the proportion under consideration and n is the total number of cells counted.We derived two normal female human induced pluripotent stem cell (hiPSC) lines (WK1 and WK6) by standard iPS methods from dermal fibroblasts from a 3 year old female (hDF1) and 42 year old female (hDF6) donor, respectively and confirmed their pluripotency through assessment of pluripotency marker expression by immunofluorescence , fluoresAnalysis by qRT-PCR for the key pluripotency transcription factors OCT4 and REX1HLCs were generated from pluripotent stem cells cultured on matrigel in the presence of mTESR1 by the sequential exposure of pluripotent cells to cocktails of growth factors in chemically defined media . Human iHLC) and hiPSCs (WK1HLC and WK6HLC), though absolute expression levels of ALB and AFP mRNA in HLCs of hiPSC origin was lower than in HLCs of hESC origin whereas up-regulation in WK1HLCs was less prominent . ALB levels were less induced, ranging from 90 fold for WK1HLCs to 190 fold for WK6HLCs and 400 fold for WA09HLCs (data not shown). We also observed about 100 fold up-regulation of hepatocyte nuclear factor 4\u03b1 (HNF4\u03b1), a nuclear receptor transcription factor that activates expression of numerous hepatocyte-specific genes encoding ALB, apolipoproteins, and several Cytochrome P450 (CYP) enzymes with ALB in all stage 3B cultures . APOA1, iPSC line to ten-fold for HLCs derived from the WK1iPSC line Our HLCs secrete cholesterol and express high levels of HMGCR , two cliPeriportal HLCs from human iPS cells provide a powerful new platform for the study of hepatic functions that impact cardiovascular health as mediated by serum cholesterol. It is well established that human genetic variation impacts cholesterol homeostasis. Extreme examples of genetic effects include familial hypercholesterolemias, often caused by defects in the LDLR gene Lastly, our iPS cell culture model removes the confounding variable of dietary cholesterol, which is difficult to control in studies of human subjects. Our cholesterogenic HLC model thus provides a novel and effective way to examine the contribution of individualized genomic variation to cholesterol homeostasis, with obvious implications for advances in cardiovascular health research.Figure S1Morphological assessment of pluripotent stem cell lines H9, WK1 and WK6 by light microscopy. Phase contrast microscopy of pluripotent stem cell lines WA09, WK1 and WK6 grown on feeder cells with human embryonic stem cell medium (A) and on matrigel with mTESR1 (Stem Cell Technologies) (B). Both WK1 and WK6 cells show the typical colony morphology with distinct edges independent of the growth conditions on feeder cells or matrigel as was observed for WA09 embryonic stem cells. Individual iPSCs also show the large ratio of nuclear to total cell volume typical for human embryonic stem cells (WA09).(TIF)Click here for additional data file.Figure S2Directed differentiation of hiPSC line WK1 to HLCs. Immunofluorescent detection of the hepatic lineage markers SOX17 and ALB during hepatic differentiation of hiPSC line WK1. The normal human dermal fibroblast line hDF1 (row 1) was reprogrammed to yield hiPSC line WK1 (row 2), WK1, was subjected to the three-stage directed differentiation procedure outlined above (ed above . Undiffe(TIF)Click here for additional data file.Table S1List of qRT- and RT-PCR primer sequences used in this study.(DOCX)Click here for additional data file.Table S2Regulation of gene expression for selected genes during hepatic differentiation of WA09 hES cells.(DOCX)Click here for additional data file.Table S3Regulation of gene expression for selected genes during hepatic differentiation of WK1 iPS cells derived from hDF1 fibroblasts.(DOCX)Click here for additional data file.Table S4Regulation of gene expression for selected genes during hepatic differentiation of WK6 iPS cells derived from hDF6 fibroblasts.(DOCX)Click here for additional data file."} +{"text": "Global tissue hypoxia portends poor outcomes. Detection and prompt interventions designed to counter the effects of inadequate tissue perfusion are paramount. The relationship between venous saturations in different venous pools remains elusive and dependent upon the patient\u2019s hemodynamic status.SVCO2) and pulmonary artery (MvO2) at three different time points (prior to operation (T0), immediately after weaning from CPB (T1) and on postoperative day 1 (T2)) from 89 consecutive cardiac surgical patients. Thermodilution cardiac indices and serum lactate measurements were obtained. Clinical outcomes were monitored.Venous samples were drawn from the superior vena cava . Patients with a larger postoperative negative MvO2-SSVCO2 gradient had longer CPB and cross-clamp times . This did not correlate with inferior clinical outcomes or laboratory markers of hypoperfusion.The difference between the MvO2-SSVCO2 gradient. Widening of the gradient between MvO2 and SSVCO2 in favor of the latter correlated with the duration of the CPB and ischemic times. Extrapolating functional cardiac performance based on SSVCO2 would be unreliable in this setting, with greater errors seen following more complex operations.The relationship between the venous saturations between different venous pools was inconsistent over the course of the immediate postoperative period, with a tendency towards expansion of the negative MvO"} +{"text": "Left-ventricular (LV) shape remodeling has known association with cardiac pathophysiology and dysfunction. While a prolate spheroid shape is vital for optimal cardiac function, LV dilatation and high sphericity is characteristic of cardiomyopathy. Early quantification of abnormal LV morphology may potentially guide clinical decisions to check disease progression. However a quantitative biomarker of 3D LV morphometry is found wanting in routine clinical practice as an upgrade to the rudimentary sphericity index.Spherical harmonics (SPHARM) shape descriptors were sought to quantify patient-specific LV shape. Morphometric variation from a normal LV shape was studied with application to diagnostic identification of cardiomyopathy. Cardiac MR (CMR) was employed for anatomical examination and end-diastolic LV endocardial surface models were segmented from short-axis SSFP cine scans of 11 pediatric subjects between the ages 2 and 17. Surface models were individually registered by rigid transformation and scaling to match the LV base diameter of a reference 17 year old normal LV that approximated well to a prolate spheroid. Two characteristic morphometric difference percentages (DP) were computed as a percentage of normal LV length for each case with respect to the reference normal model i.e. maximum DP (MDP) and average DP (ADP). A 16 patient cohort was used for training, comprising 10 simulated normals (ADP < 3.5 % & MDP < 13%) and 6 abnormal cases including 2 with LV hypertrophy (LVH), 3 with arrythmogenic right ventricular dysplasia (ADVD), 1 post myocardial infarction (MI). Rule-based decision trees were prepared to predict a response function defining normal vs. pathological cases using the new MDP and ADP predictors simultaneously with the conventional LV sphericity index as a third predictor, through a 3D generative clustering approach. Classification accuracy was evaluated with a 6 patient test cohort comprising 3 ARVDs, 2 LVH and 2 normal subjects.The underlying principle of this LV-base normalized diffeomorphic analysis is that LV remodeling is accompanied by heightened sphericity. This assumption was verified by the existence of positive Spearman rank correlations for sphericity with ADP (0.41) and MDP (0.28). 100% test classification accuracy was obtained using a simultaneous thresholds for MDP (>13.9%), ADP (>3.5%) and sphericity (>0.7). ARVD patients were all identified as pathologies based upon LV shape alone. Further, strong correlation (0.94) was observed between LV sphericity and age for ARVD patients.This pilot study suggests DP is a valuable biomarker in identifying cardiomyopathy. Strong correlation between sphericity and ARVD patient age combined with the excellent classification of ARVD as a cardiomyopathy by LV shape alone supports possible left-sided involvement in ARVD. Analysis with a larger cohort comprising more normal controls is warranted to validate this hypothesis.No funding sources to disclose."} +{"text": "Cardiac MRI (CMR) is increasingly recognized as a valuable imaging modality for the investigation of patients with suspected acute coronary syndrome found to have non obstructive coronary arteries on angiography. Studies have shown that early CMR within 48 hours can facilitate diagnosis of important differential diagnoses, particularly myocarditis. We attempted to demonstrate whether later scanning, after 6 weeks, would still have a similar pick up rate for these diagnoses, compared with earlier CMR within 6 weeks.64 patients with suspected acute coronary syndrome referred for coronary angiography at a tertiary cardiac centre who had normal coronary arteries enrolled to the study and were entered into a prospective registry. All underwent cardiac MRI. Axial black blood, multiplanar cines, STIR and delayed enhancement sequences were taken. The first 32 patients had cardiac MRI performed after 6 weeks, and the subsequent 32 had MRI performed within 6 weeks. The two groups were compared with respect to diagnoses made on MRI, as well as demographics and baseline troponin concentrations.32 patients underwent CMR scanning after 6 weeks, at an average of 50.5 days. The final diagnoses were myocardial infarction (36%), myocarditis (13%), Takotsubo cardiomyopathy (6%) and non-diagnostic (45%). The subsequent 32 patients underwent CMR within 6 weeks, at an average of 22.6 days. The final diagnoses in this group were myocardial infarction (19%), myocarditis (41%), Takotsubo cardiomyopathy (13%), non-diagnostic (25%) and hypertrophic cardiomyopathy (single case).Outpatient CMR performed on patients with suspected acute coronary syndrome but with normal coronary arteries later than 6 weeks has a lower diagnostic yield compared to early outpatient CMR within 6 weeks. The pick up rate for myocarditis on CMR within 6 weeks is comparable to that in previous studies of CMR performed within 48 hours of admission, suggesting that it is feasible to perform scans during this time without compromising diagnostic yield.No external funding was received."} +{"text": "For self-renewal, embryonic stem cells (ESCs) require the expression of specific transcription factors accompanied by a particular chromosome organization to maintain a balance between pluripotency and the capacity for rapid differentiation. However, how transcriptional regulation is linked to chromosome organization in ESCs is not well understood. Here we show that the cohesin component RAD21 exhibits a functional role in maintaining ESC identity through association with the pluripotency transcriptional network. ChIP-seq analyses of RAD21 reveal an ESC specific cohesin binding pattern that is characterized by CTCF independent co-localization of cohesin with pluripotency related transcription factors Oct4, Nanog, Sox2, Esrrb and Klf4. Upon ESC differentiation, most of these binding sites disappear and instead new CTCF independent RAD21 binding sites emerge, which are enriched for binding sites of transcription factors implicated in early differentiation. Furthermore, knock-down of RAD21 causes expression changes that are similar to expression changes after Nanog depletion, demonstrating the functional relevance of the RAD21 - pluripotency transcriptional network association. Finally, we show that Nanog physically interacts with the cohesin or cohesin interacting proteins STAG1 and WAPL further substantiating this association. Based on these findings we propose that a dynamic placement of cohesin by pluripotency transcription factors contributes to a chromosome organization supporting the ESC expression program. The cohesin complex, consisting of four core subunits ), is important for a variety of biological processes including chromosome segregation, DNA-damage repair and chromosome morphology Recent studies have extended the canonical functions to a role of cohesin in gene regulation Interestingly, we and others recently identified cohesin subunits in RNAi ESC screens as factors that are required to maintain Oct4 expression p-value of 10e-5, we identified 15311 specific RAD21 binding sites employing a high-affinity GFP-antibody. Applicability and specificity of the BAC tagging approach for generic assays has been demonstrated previously ng sites , Fig. S2Consistent with genome-wide binding studies for other cohesin subunits in vivo.Close inspection of the RAD21 and CTCF peaks revealed that RAD21 typically does not exactly overlap with the CTCF binding position. Instead, RAD21 peaks were slightly shifted in respect to the CTCF summits . For a cin vivo, we compared our data with published ChIP-seq data Despite the high overlap of RAD21 and CTCF binding, we also identified 4087 (26.7%) RAD21 binding sites that did not co-localize with CTCF in ESCs . This reThe co-localization of RAD21 with pluripotency transcription factors prompted us to further investigate the functional role of RAD21 and other cohesion subunits in ESC maintenance. Because of its essential role during cell division, complete removal of cohesin causes cell death To test whether the expression change of pluripotency and early developmental genes in RAD21 depleted cells is also valid for other cohesin subunits, we repeated the expression analysis with SMC1a and SMC3 depleted ESCs and detected similar expression changes of pluripotency and lineage markers . Althoug15) , manifesTo place the observed expression changes into context of other factors, we compared the RAD21 expression profile to published data Functional relevance of RAD21 co-localizing with pluripotency maintaining transcription factors for ESC identity would suggest that these binding sites are ESC specific and that RAD21 would not be found there when cells exit the pluripotency program. To investigate this assumption, we differentiated LAP-tagged RAD21 ESCs into embryoid bodies (EBs) and performed ChIP-seq using the same parameters as employed previously for ESCs. With this analysis, we detected 11022 RAD21 binding sites in EBs. Interestingly, around half of the 15311 binding sites detected in ESCs, namely 7012 (45.8%), were also detected in EBs. This result suggests that many of the RAD21 binding sites are maintained upon differentiation . In suppBecause cohesin by itself does not seem to bind DNA sequence specifically, other mechanisms to place it at specific regions in the genome must exist. For RAD21 binding at CTCF sites, CTCF is suggested to localize cohesin via protein-protein interaction with STAG (SCC3) \u22122) to calculate the number of common cohesin binding sites RAD21 binding sites that exhibited enrichment for pluripotency related transcription factors binding sites. The fact that these sites disappear in EBs and new binding sites emerge, which are enriched for early developmental transcription factors suggests that these sites are functionally relevant for the execution of developmental expression programs. The expression changes observed after RAD21 depletion mirroring knockdown of Nanog further support this notion.Notably, the number of CTCF independent RAD21 binding sites in ESCs is considerably lower than reported for other cohesin subunits, namely SMC1a and SMC3 With several ChIP-chip and ChIP-seq data sets published A remaining question is how the transcription factors interact with the cohesin proteins implicated in chromosome conformation. The study of Kagey and colleagues showed interaction of the cohesin loading factor NIPBL with the mediator complex Collectively, these data suggest a complex interplay of the different co-localizing protein complexes and strongly indicate that these factors act in concert to maintain embryonic stem cell identity. However, a possible hierarchy of events and functions for the cooperativity of RAD21 and other cohesin subunits with pluripotency transcription factors at these sites needs future investigations. A detailed dissection of interactions between cohesin proteins and pluripotency transcription factors will certainly enhance our understanding of transcriptional regulation in light of higher order chromosome architecture.Feeder-free mouse R1/E ESC were maintained on gelatin-coated dishes in DMEM media supplemented with 15% FBS (Gibco), 0.055 mM \u03b2-mercaptoethanol (Gibco), 1\u00d7 MEM non-essential amino acids (Invitrogen), 5000 u/ml penicillin-streptomycin (Invitrogen) and 16 ng LIF , RP23-236B2 (CTCF) and RP24-230P19 (Nanog)). A LAP cassette BACs harbouring the genes of interest were obtained from the BACPAC Resource Center and blotted on nitrocellulose membranes (Millipore). Membranes were probed with antibodies against GFP , SMC1 and SMC3 and RAD21 . GAPDH was used as a loading control.Three days post RNAi, cells were fixed with 4% paraformaldehyde (Sigma), rinsed in PBS and stained using Alkaline Phosphatase Red Microwell substrate (Sigma). Images were acquired with a Canon Power Shot G11 digital camera on the Olympus CKX41 microscope.Based on the staining intensity, percentages of differentiated, half-differentiated and undifferentiated cells were determined by counting.12\u201318 and Superscript III kit (Invitrogen). Quantitative real-time PCR analysis was carried out using SYBR green master mix (Abgene) and the MxP3000 detection system (Stratagene). Samples were run in triplicate and transcript levels were calculated according to the \u0394\u0394ct method with normalization to CyclophilinB. Primer sequences are listed in For isolation of total RNA the RNeasy Mini Kit (Qiagen) was used according to the manufacture's protocol including a DNaseI digest. Reverse transcription of 0.5\u20131 ug RNA was performed using Oligo-dTImmunoprecipitation reactions were carried out with nuclear extracts from Nanog-LAP ESC as well as wildtype ESC and subjected to shotgun mass spectrometry as previously described Cells were harvested two days post RNAi and 250 ug of isolated total RNA was labeled with the One-Cycle Target Labeling and Control Reagent Package (Affimetrix) as described in the manufacture's instructions. Probes from 4 biological replicates of RAD21 and Luc RNAi were hybridized on Mouse Genome 430.2.0 arrays (Affimetrix). Image data were analyzed with the GeneChip Operation Software applying Affimetrix default settings. Expression changes were determined by a parametric analysis of variance (ANOVA) after RMA normalization with respect to the probe GC content using Partek Genomics Suite 6.4 (6.09.0129) .Gene Ontology analysis was performed using GenCoDis 2.0 For calculation of Pearson's correlation factor, published microarray data were used RAD21-LAP ESC were crosslinked with 1% formaldehyde for 10 min at room temperature and crosslinking was quenched with 125 mM glycine. Sonicated chromatin with an average size of 500 bp was immunoprecipitated over night using a GFP-antibody (MPI-CBG antibody facility) and control IgG-antibody (Dianova) and immobilized on G-sepharose . Specificity of the GFP-antibody has been validated before and sequenced with the Genome Analyzer II (Illumina).p-value threshold of 10\u22124, and with the following settings for the previously generated data \u22125. Prior peak detection, ChIP-seq fragment coordinates from CTCF, Pou5f1, Nanog, Sox2, Klf4 and Esrrb ChIP-seq data liftOver tool Peak calling was performed using MACS 1.4beta http://pinkthing.cmbi.ru.nl) software was used for assigning sites to the nearest gene and to determine genomic distribution of identified binding sites. Peak locations were visualized using the University of California Santa Cruz (UCSC) genome browser.PINKTHING accessed from Ensembl.http://code.google.com/p/bedtools) and used as a basis for further search of occurrences of known transcription factor binding motifs from the JASPAR database p-value not exceeding 10\u22125 were considered significant. Gaussian mixture modeling of distances between binding sites and motifs was performed with R package mclust .Sequences from a range of 150 bp around peak summits were extracted with BEDTools package . Peaks that were not bound by CTCF and did not contain the CTCF binding motif in the 150 bp vicinity of the peak summits were considered motif and CTCF binding independent binding sites (MCIB). The binding site and the motif enrichment MIAME compliant microarray and sequencing data from this study have been deposited in the MIAME compliant database GEO with the accession number GSE24030.Figure S1Validation of BAC-GFP tagged ESC lines and GFP-antibody for ChIP-sequencing. (A): Immunostaining of RAD21 and CTCF BAC-GFP ESC stably expressing RAD21-LAP or CTCF-LAP confirmed nuclear localization. In the merged picture, GFP (LAP tagged protein) is shown in red, \u03b1-Tubulin in green and DNA (DAPI) in blue. Scale bar equals 20 \u00b5m. (B): Western blot analysis of RAD21 and CTCF BAC- GFP ESC depleted in RAD21 and CTCF, respectively. Two independent esiRNAs and control esiRNA (Luc) were used. GAPDH expression served as protein loading control. (C): Western blot analysis of RAD21-LAP immunoprecipitation using GFP-antibody to confirm antibody specificity. Co-immunoprecipitation of the cohesin complex members SMC1a and SMC3 support functionality of the RAD21 BAC transgenic ESCs. (D): ChIP of RAD21-LAP cells transfected with control (Luc) and RAD21 esiRNA. Enrichment of selected MCIB sites that appertain to indicated genes was quantified by qPCR and confirmed reduced signals upon RAD21 depletion .(TIF)Click here for additional data file.Figure S2Identification of RAD21 binding sites and their genome distribution. (A): Table shows the number of identified binding sites using MACS algorithm in dependency of the p-value. Numbers are listed for RAD21 and IgG ChIP-samples. Binding sites detected with the p-value of 10e-5 were used for further analysis. (B): Genome distribution of RAD21 binding sites indicated that the majority of RAD21 sites is located in introns and far distant (>25 kb) from the transcriptional start site. (C):De novo motif analysis in the 150 bp vicinity of RAD21 peak summits did not reveal a RAD21 specific consensus sequence. Search for known DNA binding motifs in the 150 bp vicinity of peak summits using Jaspar database identified the CTCF motif to be the most abundant. (D\u2013E): CTCF ChIP-seq data analysis according to A\u2013C.(TIF)Click here for additional data file.Figure S3Motifs present in the vicinity of RAD21 binding sites apart from CTCF do not exhibit binding directionality. Motifs in the vicinity of RAD21 apart from CTCF do not exhibit directionality. The histogram plot of distances between RAD21 binding site and FoxI1 motif sequence in 5\u2032 to 3\u2032 strand direction does not exhibit directionality of RAD21 binding. Black lines are the components of the Gaussian mixture modelling distribution of the distances. Solid lines indicate the most dominant distributions. The red bar indicates RAD21 binding site and the black ellipses indicate expected positions of FoxI1 binding sites.(TIF)Click here for additional data file.Figure S4Depletion of cohesin strongly changes ESC morphology. ESC transfected with esiRNAs against RAD21 (2 independent esiRNAs), SMC1a and SMC3 exhibit strong change in morphology 72 h post RNAi compared to non-targeting control (Luc). Depletion of Nanog served as a positive control for ESC differentiation. Scale bars correspond to 100 \u00b5m.(TIF)Click here for additional data file.Figure S5Depletion of SMC1a and SMC3 reflects expression changes upon RAD21 knock-down. (A): Alkaline phosphatase staining of ESCs, which had been transfected with secondary esiRNAs targeting RAD21, SMC1a and SMC3 (72 h post RNAi). Nanog depletion and a non-targeting control (Luc) served as a positive and negative control for ESC differentiation, respectively. Scale bars correspond to 100 \u00b5m. (B+C): qPCR result of detected expression changes in (B) stem cell maintenance genes and (C) lineage marker genes upon knock-down of SMC1a and SMC3 versus a Luc control .(TIF)Click here for additional data file.Figure S6Validation of microarray gene expression results. Diagram shows qPCR based confirmation of up- and downregulation of selected developmental and stem cell maintenance related genes identified in the RAD21 microarray gene expression array Click here for additional data file.Figure S7Re-analysis of SMC1 and SMC3 ChIP-seq data. Analysis of the SMC1, SMC3 and CTCF DNA binding data. \u22125). Numbers of binding sites are presented for the individual experiments, calculating intersections of each of the two replicates to define CTCF overlapping and independent binding sites. The green dashed line separates cohesin sites that are independent of CTCF binding from sites that are independent of CTCF binding and do not contain the CTCF motif sequence .(TIF)Click here for additional data file.Table S1RAD21 binding sites in ESCs and EBs.(XLS)Click here for additional data file.Table S2Enrichment analysis of transcription factor binding motifs at RAD21 binding sites in ESCs and EBs.(XLS)Click here for additional data file.Table S3Enrichment analysis of RAD21 co-localizing with ESC transcription factor binding sites in ESCs.(XLS)Click here for additional data file.Table S4Gene expression microarray data of RAD21 depleted ESCs.(XLS)Click here for additional data file.Table S5List of Nanog interacting proteins identified by mass spectrometry.(XLS)Click here for additional data file.Table S6Primer sequences for esiRNA production, qPCR and ChIP.(XLS)Click here for additional data file.Table S7High-confidence cohesin binding sites.(XLS)Click here for additional data file."} +{"text": "Natriuretic peptides have potent natriuretic and vasodilator properties, reduce sympathetic drive, inhibit aldosterone secretion and have antiproliferative and antihypertrophic properties . IncreasLCZ696 is a first-in-class angiotensin receptor neprilysin inhibitor (ARNI) which provides concomitant inhibition of NEP and the angiotensin receptor. Ingestion of LCZ696 delivers systemic exposure to AHU377 and to valsartan (an angiotensin II receptor blocker). LCZ696 treatment is associated with dose-dependent increases in plasma cGMP, renin activity and angiotensin II, consistent with the drug\u2019s dual mechanism of action; a dose of 200-400 mg LCZ696 achieves approximately 90% of its maximal NEP inhibition [P<0.05); 24-hour ambulatory BP monitoring results were consistent with office BPs. Pulse pressure was also lowered to a greater extent with LCZ696 than the respective valsartan groups (both P<0.05) [P<0.01) as compared to baseline values (4). The Phase III PARADIGM-HF study is evaluating whether LCZ696 is superior to enalapril in delaying the time to cardiovascular mortality or first occurrence of heart failure (HF) hospitalization in patients with HF and reduced ejection fraction [In a large Phase 2 study in hypertensive patients n=1328), LCZ696 400mg and 200mg once daily reduced mean office systolic and diastolic blood pressure (BP) by an additional 6/3 and 5/3 mmHg compared with valsartan 320 and 160 mg, respectively (both 28, LCZ69Dual inhibition of neprilysin and the angiotensin receptor with LCZ696 may represent an attractive therapeutic approach for a range of cardiovascular diseases, including hypertension and heart failure, in which vasoconstriction, volume overload and neurohormonal activation play a part in pathophysiology."} +{"text": "The induction of an efficacious anti-tumor immune response (IR) requires the cross-processing and presentation of tumor antigen by Dendritic Cells (DCs). Block of endocytated tumor associated antigen (TAA) in the early compartments of the intracellular processing machinery shifts the IR towards a Th2 balance. MUC1 is one of the most relevant tumor associated glycoprotein expressed by epithelial cells and its immunogenicity is altered by the glycosylation profile. Moreover soluble MUC1 antigen has shown to be stucked in endolysosomal compartment of DCs thus inducing mostly a Th2 response. Objective of this study was to investigate whether glycosylation pattern and MUC1 bound to microvesicles could influence the antigen processing by DCs. MUC1 as soluble molecule, independently by the glycosylation profile, appears to be blocked in the pre-endosomal compartment. Receptor-mediated endocytosis pushes further the processing in the HLAII compartment. Cross-processing of MUC1 in HLAI compartment is observed only when MUC1 is carried by microvesicles (MUC1-MVs). Moreover only DCs stimulated with MUC1-MVs are able to induce IFN-\u03b3 production by MUC1 specific CD8+T cells. The distinct processing of the MUC1 membrane bound is accompanied by deglycosylation processes thus generating Tn-MUC1 immunogenic glycoforms. These results show that MUC1 undergoes to alternative processing pathways depending by its form of release (MVs vs soluble) thus modifying MUC1 immunogenicity. Moreover these experimental evidence can be important to design efficacious glycoantigen formulations for DC-based cancer vaccines."} +{"text": "We report the draft genome sequences of 10 proteobacterial strains isolated from beach sands contaminated with crude oil discharged from the Deepwater Horizon spill, which were cultivated under aerobic and anaerobic conditions with crude oil as the sole carbon source. All strains contain multiple putative genes belonging to hydrocarbon degradation pathways. Acinetobacter sp. strain COS3, Alcanivorax sp. strain P2S70, Marinobacter sp. strain C1S70, Labrenzia sp. strain C1B10, and Marinobacter sp. strain C1B70) or under anaerobic conditions with nitrate as the sole electron acceptor (in situ in the same contaminated sands (The Deepwater Horizon (DWH) well blowout resulted in the largest accidental release of oil into the marine environment to date . The ultin PBN3) . Our preed sands . Oil deged sands . The iniAlphaproteobacteria and Gammaproteobacteria . The contigs were successfully used for annotation and gene prediction by RAST represent the first genome sequences of the group for strains grown on crude oil.The draft genome sequences were assembled in order to understand the metabolic potential for degrading crude oil components at the strain level, as well as to provide reference genomes in support of metagenomic work to characterize microbial populations in beach sands contaminated during the Deepwater Horizon discharge. Furthermore, we seek to compare the underlying genetic basis for hydrocarbon degradation across diverse bacterial strains competing in the same temporal and spatial environment. Substantial variation in the assembly and number of hydrocarbon degradation genes among strains of the same genus (AXBX00000000 (Alcanivorax PN-3), AXBZ00000000 (Alcanivorax P2S70), AXCC00000000 (Marinobacter EN3), AXBV00000000 (Marinobacter ES-1), AXCB00000000 (Marinobacter EVN1), AXBW00000000 (Marinobacter C1S70), AXCD00000000 (Acinetobacter COS3), AXCA00000000 , AXBY00000000 (Labrenzia C1B10), and AXCE00000000 (Labrenzia C1B70).The draft genome sequences of the strains obtained in this study have been deposited in GenBank as part of BioProject no. 217943, with individual genome sequences submitted as whole-genome shotgun projects in GenBank under accession no."} +{"text": "To determine the ability of adenosine stress perfusion cardiac magnetic resonance (CMR) to accurately identify the culprit vessel.The diagnostic evaluation of patients with suspected ischaemic heart disease (IHD) frequently involves a functional assessment of ischaemia. Adenosine stress perfusion CMR is a non-invasive test with high sensitivity for the detection of IHD, however rarely is its accuracy for culprit vessel identification assessed. We sought to determine the accuracy of stress perfusion CMR for identifying the culprit vessel. Furthermore, we sought to affirm the specificity of a positive CMR against an angiographic gold standard.Retrospective study of patients with a positive stress perfusion CMR and subsequent coronary angiography. Perfusion imaging was obtained at stress (adenosine 140 \u00b5g/kg/min) and rest on a 1.5T scanner. Late enhancement was assessed with dual pass gadolinium . Angiographic stenosis \u2265 50% was defined as significant. The presence or absence of a significant lesion together with the correlation between ischaemic territory on CMR and angiographic culprit vessel was evaluated.Thirty seven patients had a positive CMR with subsequent angiography, with follow up data for a median 24 months (IQR 21-27 months). Thirteen patients (35%) had previous myocardial infarction or revascularisation. Of the cohort of 37, six (16%) had normal angiograms and 31 (84%) had a significant epicardial stenosis. Of the six false positives, three had localised septal hypoperfusion, while a further three had circumferential defects. Of the 31 patients correctly identified, CMR accurately established the territory of the culprit vessel in 29 (94%). The vessels confirmed as ischaemic were left anterior descending 12 (39%), circumflex 6 (19%) and right coronary artery 13 (42%).Adenosine stress perfusion CMR reliably identifies the territory supplied by the culprit vessel (29 out of 31 - 94%). Furthermore, we reaffirm the high specificity (84%) of stress perfusion CMR."} +{"text": "Glossina), the female accessory gland is specialized to generate nutrients in the form of a milk-like secretion to support growth of intrauterine larva. Multiple milk protein genes are expressed specifically in the female accessory gland and are tightly linked with larval development. Disruption of milk protein synthesis deprives developing larvae of nutrients and results in extended larval development and/or in abortion. The ability to cause such a disruption could be utilized as a tsetse control strategy. Here we identify and delineate the regulatory sequence of a major milk protein gene (milk gland protein 1:mgp1) by utilizing a combination of molecular techniques in tsetse, Drosophila transgenics, transcriptomics and in silico sequence analyses. The function of this promoter is conserved between tsetse and Drosophila. In transgenic Drosophila the mgp1 promoter directs reporter gene expression in a tissue and stage specific manner orthologous to that of Glossina. Analysis of the minimal required regulatory region of mgp1, and the regulatory regions of other Glossina milk proteins identified putative homeodomain protein binding sites as the sole common feature. Annotation and expression analysis of Glossina homeodomain proteins identified ladybird late (lbl) as being accessory gland/fat body specific and differentially expressed between lactating/non-lactating flies. Knockdown of lbl in tsetse resulted in a significant reduction in transcript abundance of multiple milk protein genes and in a significant loss of fecundity. The role of Lbl in adult reproductive physiology is previously unknown. These results suggest that Lbl is part of a conserved reproductive regulatory system that could have implications beyond tsetse to other vector insects such as mosquitoes. This system is critical for tsetse fecundity and provides a potential target for development of a reproductive inhibitor.Regulation of tissue and development specific gene expression patterns underlies the functional specialization of organs in multi-cellular organisms. In the viviparous tsetse fly ( Glossina) harbor and give birth to live young. To do this, they nourish their intrauterine larvae with milk secretions. This work focuses upon understanding the regulation of tsetse milk proteins, which are essential for fecundity and are expressed in a temporally and spatially specific manner by pregnant females. We identified the minimal upstream regulatory DNA sequence of the major milk protein gene mgp1, which confers tissue specific expression in the female accessory glands of reproductively active flies. This regulatory sequence functions similarly in transgenic fruit flies (Drosophila melanogaster) and drives expression of reporter gene products in the adult female accessory gland. Comparison of this regulatory sequence with sequences from other characterized milk proteins indicates that conserved homeodomain transcription factors may be responsible for regulating these genes. Analysis of Glossina homeodomain proteins identified an accessory gland/fat body specific factor, Ladybird late (lbl), which appears to regulate the expression of multiple milk proteins. Reduction of lbl levels interferes with milk protein gene expression, which in turn reduces Glossina fecundity. These results suggest that milk proteins in Glossina are regulated by a conserved regulatory system mediated in part by the homeodomain transcription factor lbl. Components of this system could provide a target for development of a tsetse reproductive inhibitor.Female tsetse flies (Diptera: Glossina sp.) are the exclusive vectors of Human African Trypanosomiasis (HAT) and nagana in Sub-Saharan Africa. HAT transmission can be prevented through tsetse population control by baited targets and trapping Tsetse flies development by the mother GMOY009745) as one of the primary protein components of tsetse milk Diplotera punctatamgp1 transcript and protein levels begin to increase and reach their maximum when the larva reaches its 3rd instar mgp1 by dsRNA treatment reduces female fecundity suggesting that this protein is required for larval development Molecular analysis of milk gland secretions has identified Milk Gland Protein 1 (MGP1 - VectorBase accession:trf) asmase1) mgp2-10) the function of which is to be determined Glossina genome paper) Transcriptomic analysis of pregnant females and proteomic analysis of milk gland secretions have provided knowledge on the protein constituents of tsetse milk mgp1), focusing on the role of specific transcription factors that may be associated with the upstream promoter region. We performed our analysis of milk protein gene regulation in tsetse using a combination of classic molecular techniques and genetic analysis in transgenic Drosophila. We used in silico analysis of genomic data and multiple high throughput datasets coupled with functional studies in Glossina and in Drosophila to identify candidate homeodomain factors that are apparently involved in the regulation of multiple tsetse milk protein genes. Our results suggest conservation of a reproduction associated gene regulatory mechanism across different taxa (Glossina and Drosophila) in the context of differing reproductive physiologies. We discuss our findings in light of potential methods to disrupt the reproductive capacity of tsetse as a vector control tool.The recent sequencing of the tsetse genome qPCR primer sequences are found in Glossina morsitans morsitans colony maintained in the insectary at Yale University was originally established with puparia from fly populations in Zimbabwe. Newly emerged flies are separated by sex and mated at three to four days post-eclosion. Flies are maintained at 24\u00b11\u00b0C with 50\u201355% relative humidity, and receive defibrinated bovine blood every 48 h using an artificial membrane system The rd instar larvae were collected. Flies were checked every 24 hours and individuals that had undergone parturition during the 24 hour period were collected to stage them in sample groups. Three flies were collected at the following time points, pregnant (3rd instar larva), 0\u201324, 24\u201348, 48\u201372, 72\u201396, 96\u2013120, 120\u2013144, 144\u2013168, and 168\u2013192 hrs post parturition (pp). RNA and protein were isolated from flash frozen female flies and tissue samples utilizing the standard Trizol protocol with a modification to the final step in which the isolated protein pellets are dissolved in cracking buffer and 4% CHAPS) mgp1 were determined by CFX Connect Real Time PCR Detection System with SYBR Green Supermix . The data were analyzed with software version 3.1 (Bio-Rad). All samples were normalized according to Glossina tubulin (tub) expression levels.Samples for gene expression analysis during parturition were collected as follows. Female flies pregnant with 3Western blotting was performed utilizing the protein from the samples described above using previously described antisera and protocols G. m. morsitans genome http://gmorsitans.vectorbase.org/Glossina_morsitans/Info/Index).Genomic sequences and automated annotations were derived from the recently completed mgp1 predicted transcription start site. These fragments were PCR amplified from G. m. morsitans genomic DNA using Platinum Taq DNA Polymerase High Fidelity (Invitrogen) and sub-cloned into the T-vector cloning vector . Inserts were excised from the T-vector using SphI and NotI and ligated into the p-element vector pPelican lacZ reporter gene downstream of the regulatory sequence. The enhancer/reporter constructs were used to transform 1118w flies by p-element mediated transformation via a commercial transformation service (Best Gene: Drosophila Injection Services (http://www.thebestgene.com). The transformations generated 4 lines carrying the 2.0 kB construct mgp1-Bgal-2.0-1-4 and 3 lines carrying the 0.5 kB construct mgp1-Bgal-0.5-1-3. Drosophila lines were maintained according to standard protocols Two constructs were created using 2 kB and 0.5 kB of the 5\u2032 upstream from the Drosophila were dissected in 1\u00d7 PBS. Dissected tissues were fixed and stained for \u03b2-galactosidase activity with the \u00df-galactosidase Staining Kit following the manufacturer's protocols.Tissues from male and female mgp1-egfp-509), 236(mgp1-egfp-236), 112 (mgp1-egfp-112) or 13 bp (mgp1-egfp-13) nucleotide versions of the mgp1 upstream sequence were generated utilizing the PhiC homologous recombination transformation system mgp1 upstream fragment was PCR amplified from Glossina morsitans genomic DNA using Platinum Taq DNA Polymerase High Fidelity (Invitrogen) and sub-cloned into the T-vector cloning vector (Promega). The fragment was excised with SphI and SpeI then ligated into the nuclear EGFP enhancer analysis vector pStinger mgp1-pStinger construct was used as a template to amplify the 509, 236, 112 and 13 base pair variants of the mgp1 promoter-egfp fusion constructs. The forward and reverse primers for the constructs were tailed with the AttB40 PhiC recombination site sequence. Amplified constructs were cloned into T-vector and sent for transformation via a commercial transformation service (Best Gene: Drosophila Injection Services (http://www.thebestgene.com). Constructs were injected into the Drosophila line genotype y1 w*; P[attP.w+.attP]JB53F (Bloomington Stock Center #27386). The AttB integration site is 53F8, 2R:12985015 nd chromosome balancer flies and screened for loss of eye color as a negative marker of transgene insertion. Drosophila lines were maintained according to standard protocols Flies expressing an eGFP reporter with 509 . Data was analyzed using software version 3.1 (Bio-Rad). All treatments were normalized according to Drosophila beta-tubulin expression levels using gene specific primers and carried out in triplicate. Visual inspection of transgene expression was performed by tissue dissection in 1\u00d7 PBS followed by fluorescent microscopy using a Axio Cam dissecting scope equipped with a X-cite series Q fluorescence system .RNA was isolated from flies using Trizol (Invitrogen). cDNA was prepared from total RNA using the Invitrogen Superscript III kit (Invitrogen). Transgene transcript levels were quantified by qPCR with iQ SYBR Green Supermix using egfp transgene expression was performed as described above with total RNA from 3 independent groups of larval, pupal, and 3\u20135 day old male and female flies from the mgp1-egfp-509 line. 3\u20135 day old adult females from the mgp1-egfp-236, mgp1-egfp-112 and mgp1-egfp-13 lines were also collected and analyzed in the same manner to compare transgene expression between the lines.Stage specific quantification of mgp1-egfp-509 line on either complete or minimal media in 25\u00d795 mm polystyrene vials. Egg deposition was monitored daily for each group. Parallel treatments were run to quantify egfp levels in stressed flies using the qPCR methods described above.Nutritional deprivation experiments were performed by maintaining the mgp upstream regulatory region using the \u201cMatInspector\u201d program from Genomatix (http://www.genomatix.de/) Glossina genome (www.vectorbase.org) mgp1 (GMOY009745) critical region, trf (GMOY004228), asmase1 (GMOY002246), mgp2-10 genes. 500 bp from the predicted transcription start site of each gene was used in a comparative analysis. This analysis was also performed using MatInspector.Putative transcription factor binding sites were predicted within the Glossina homeodomain sequences were identified from the Glossina genome Drosophila homeodomain factors were used to search the Glossina genomic assembly and the de novo assembly from \u221210 were collected and searched against the NCBI database by BLASTx search. Sequences were then annotated with nomenclature from the highest scoring hit result.Predicted Glossina homeodomain proteins in lactating and dry flies. RNA-seq data were mapped directly to the Glossina homeodomain genes utilizing CLC Genomics Workbench . Normalized gene expression was calculated as RPKM We have recently completed a transcriptome study focusing on differences between pregnant/lactating or dry flies (non-lactating) Glossina homeodomain genes were analyzed by qPCR based tissue and stage specific expression analysis. Tissue-specific samples were acquired from females harboring 2nd instar larva. qPCR was conducted as before and transcript levels were normalized to tubulin.Five candidate Glossina ladybird late ortholog. siRNAs were designed by web-based tools and ordered commercially (IDT). The siRNA consists of two Duplex sequences for lbl. Control siRNA molecules were designed as complementary to green fluorescent protein (GFP) mRNA . Concentration of each siRNA was adjusted to 700\u2013750 ng/\u00b5l in PBS with a Nanodrop spectrophotometer. Mated female flies were injected with 1.5 \u00b5l siRNA 6\u20138 d after adult emergence. siRNA was shown to have no discernible effect on larval transcripts tubulin) 5 d after injection for lbl and 8 d after injection for mgp1.Functional analysis of the ladybird late homeodomain factor was performed using synthesized siRNAs homologous to the mgp1 gene is related to intrauterine larval development. Previous expression data for mgp1 was based upon samples staged by female reproductive physiology and larval development mgp1 transcripts appear to decrease after larval deposition (parturition) and increase again at the onset of intrauterine larval development in the next pregnancy cycle. To obtain a high resolution temporal analysis of milk protein gene expression during this period, pregnant flies were collected and synchronized by time of parturition in 24 hour intervals rather than by pregnancy status. Transcript levels from these samples were quantified by qPCR analysis, and protein expression was determined by Western analysis. The results show that mgp1 transcript abundance and protein levels undergo a precipitous drop after parturition and reach their lowest levels at 24\u201348 hours post-parturition suborder.Analysis of the regulation of \u03b2-galactosidase reporter gene and the promoter/reporter fusion is flanked by gypsy insulator sequences to prevent the influence of local regulatory elements. Transformation was accomplished by p-element transposition. Reporter expression was visualized by staining for \u03b2-gal activity in dissected transgenic Drosophila tissues. Two constructs were created including either 2 kB or 0.5 kB sequence of the 5\u2032 upstream from the predicted mgp1 transcription start site and three transgenic lines for the 0.5 kB construct .The transformation constructs were developed using the pPelican enhancer/reporter plasmid art site . Transfomgp1 expression is specific to the milk gland of adult female flies mgp1-\u03b2-gal-2.0 and mgp1- \u03b2-gal-0.5 lines with \u03b2-gal staining to understand the sex and tissue specific nature of the reporter gene expression. Transgene expression was exclusive to the accessory glands (paraovaria) of the female reproductive tract in both transgenic lines reporter gene from the pStinger p-element transformation vector. We generated four transgenic lines, which include either 509 (mgp1-egfp-509), 236 (mgp1-egfp-236), 112 (mgp1-egfp-112) or 13 bp (mgp1-egfp-13) segments corresponding to the mgp1 regulatory region upstream of the start site females from the ng lines . Of the egfp expression during different developmental and sexual stages in mgp1-egfp-509 reveals that the transgene is only significantly expressed within sexually mature adult females with only background levels present in larvae and males , OVO transcription factor (DOVO: 1 site), Iroquois factor group (IRXF: 1 site), Tailless (DTLL: 1 site), Abdominal B (ABDB: 1 site), Paired homeodomain factors (PRDH: 2 sites), Dead Ringer (DRIF: 1 site), Giant (DGTF: 1 site) and Heat shock factors (DHSF: 1 site).We next set out to predict transcription factor binding sites that may lie within the 0.5 kb promoter region of Glossina genome mgp1, mgp2-10, trf and asmase1mgp1 region critical for transgene expression in Drosophila with the 500 bp sequence upstream of the transcription start site of these genes predicts that only homeodomain protein binding sites (DHOM) are common between all 12 sequences . The onlGlossina bearing homology to the 106 annotated Drosophila homeodomain proteins. This was accomplished by tBLASTn search of the Glossina genome assembly de novo assembly of RNA-seq libraries with Drosophila homeodomain protein sequences. All significant hits were submitted to a BLASTx comparison against the NCBI protein database. This analysis predicted a total of 96 putative tsetse homeodomain protein orthologs within the current genome assembly (see Table S36 in the Glossina genome paper).To identify homeodomain type regulatory factors that may recognize the conserved DHOM sites, we annotated and collated all sequences in nubbin (nub \u2013 VectorBase accession: GMOY012175), teashirt (tsh \u2013 VectorBase accession: GMOY011052) and vismay (vis \u2013 VectorBase accession: GMOY000857) and 2 with lower transcript abundance ladybird late (lbl \u2013 VectorBase accession: GMOY004068), and pox meso (poxm \u2013 VectorBase accession: GMOY002525) .mgp1 (and the other milk proteins), homeodomain factors specific to that tissue were of primary interest. We performed a qPCR based expression analysis of the five candidate factors using RNA from different tissue samples obtained from lactating female flies. The expression patterns were unique for each gene. Of the genes analyzed, only the Ladybird Late gene (lbl) displayed a milk gland/fat body specific expression pattern. The fat body and milk gland are grouped together due to their interconnected nature which makes separating them by dissection next to impossible. Transcripts for lbl were \u223c30 fold higher in the milk gland/fat body tissue than in any other tissue . The siRNA injections resulted in significant reductions in target gene transcript abundance. The levels of lbl were reduced to \u223c30% of controls have modified glands which produce 2 substances (one from the left and one from the right gland) which combine in the oviduct to generate an ootheca that encases their eggs and hardens into a protective coating Rhodnius prolixus) uses these glands to produce an adhesive secretion to glue eggs to substrate In tsetse, the milk gland has been a recent focus of study; however less is known regarding the function(s) of female accessory glands in Glossina genome sequence in silico analyses led us to identify the homeodomain family of transcription factors and allowed us to narrow down our search from a field of 96 potential factors to a handful of 5 factors. Homeodomain proteins are a conserved family of transcription factors characterized by the presence of a helix-turn-helix DNA binding domain called the homeodomain The recent release of the Drosophila. The conservation of regulatory sequence function in tsetse and Drosophila suggests that LbL performs orthologous reproductive functions in both species.Here, tissue specific expression profiling and RNAi analysis led us to the Ladybird Late homeodomain factor (Lbl). This factor appears critical to the expression of multiple milk protein genes in tsetse based upon our knockdown experiments. The 236 bp tsetse promoter region was sufficient to drive the synthesis of a reporter gene in an accessory gland specific manner in female lbl gene was first discovered with its paralog ladybird early (lbe) during a search for novel homeodomain proteins ladybird genes in Drosophila have linked these genes with embryonic heart cell precursor diversification ladybird gene regulation suggests that the two genes are regulated by the wingless (wg) signaling pathway and may in turn regulate wingless expression in a feedback loop lbl with wingless signaling during development hints at the possibility of the wingless pathway as a regulator of reproduction associated gene regulation in adults The mgp1 regulatory sequence between tsetse and Drosophila coupled with the ease of transgenesis in the fruit fly system allows us to use Drosophila as a surrogate in which to study female accessory gland gene expression. This promoter in combination with GAL4 mediated systems can enable in vivo tissue and stage specific knockdowns and/or ectopic expression experiments in Drosophila in order to identify components of the signaling system regulating Lbl activity. Flies carrying mgp1-GAL4/mgp1-EGFP fusions can be crossed with the available UAS-RNAi lines wg pathway) to perform knockdowns quickly and efficiently. The presence of the mgp1-EGFP reporter will allow for rapid visual assessment of knockdown phenotypes. The findings from Drosophila can then be translated back into tsetse by RNAi experiments.The similarity in function of the Glossina is essential for reproductive success and provides an important molecular target for the development of an insect specific reproductive inhibitor. Further analysis of mechanisms governing this system will provide important data towards controlling sleeping sickness as well as broadening the understanding of insect reproductive physiology. The knowledge gained from this work can be put towards the development of an insect specific inhibitor which disrupts milk protein production in tsetse. A substance such as this could be utilized on targets, traps and livestock as a non-toxic alternative to pesticides to reduce tsetse populations and decrease the trypanosome transmission threat.These findings are an important step in identifying the pathways, factors and signals regulating milk gland/accessory gland function in insect reproduction. Milk production in Figure S1Schematic of mgp1-reporter fusion constructs for Drosophila transformation.A. 2.0 and 0.5 kB mgp1 pPelican based transformation constructs. Constructs include 2.0 and 0.5 kB promoter fragments driving a \u03b2-gal reporter flanked by gypsy insulator sequences. The construct also includes the white transformation marker and is flanked by p-element sequences. B. 509,236,112 and 13 bp mgp1 pStinger based recombination constructs. The 0.5 kb mgp1 fragment was cloned into the pStinger transformation plasmid. pStinger is similar to pPelican with the exception that the \u03b2-gal reporter has been replaced with nuclear EGFP. Recombination constructs were amplified with primers containing AttB sites which amplify different lengths of the mgp1 promoter. Amplified fragments were cloned into the T-vector PCR cloning vector and used for recombinatorial transformation.(TIF)Click here for additional data file.Figure S2Tissue specific expression analysis of five putative homeodomain genes. All qPCR analyses normalized to Tsetse tub. Error bars represent standard error. Samples represent 3 replicates of tissues isolated from 5 individuals. A. qPCR analysis of nubbin (nub) tissue specificity. B. qPCR analysis of teashirt (tsh) tissue specificity. C. qPCR analysis of vismay (vis) tissue specificity. D. qPCR analysis of ladybird late (lbl) tissue specificity. E. qPCR analysis of pox meso (poxm) tissue specificity.(TIF)Click here for additional data file.Table S1qPCR primer sequences. Sequences for Syber green primers utilized in the qPCR analyses performed in this work.(DOCX)Click here for additional data file.Table S2Construct creation primer sequences. Primer sequences utilized in the amplification of sequences for the creation of transgenic constructs.(DOCX)Click here for additional data file.Table S3siRNA sequences. Sequence information for siRNAs utilized within the gene knockdown experiment.(DOCX)Click here for additional data file."} +{"text": "Arrhythmia recurrence following acutely successful catheter ablation for atrial fibrillation (AF) is associated with recovery of conduction across previously complete circumferential or left atrial (LA) linear lesions. Late gadolinium enhancement (LGE) magnetic resonance imaging (MRI) may be able to provide a non-invasive means to identify gaps in ablation lesions. This study assessed this prospectively in patients undergoing repeat LA catheter ablation.Eleven patients who had previously undergone one or more LA catheter ablation procedures for AF, and who represented with either paroxysmal AF (n=4) or atrial tachycardia (AT) (n=7) underwent LGE MRI 2-3 weeks prior to repeat catheter ablation. The scans were not analysed before repeat ablation and the procedure was performed without using any of the anatomical or scar information from the LGE MRI.After ablation, three expert blinded reviewers (who were not operators in the repeat procedure) independently scored 3D left atrial (LA) LGE MRI reconstructions Figure for the From the LGE MRI scans, interobserver agreement for detection of any gap was 95%. The positive predictive value of LGE MRI for gap detection was 35%, whilst the negative predictive value was 43%. Specificity and sensitivity were 50% and 28% respectively.There was a very high degree of agreement between the three reviewers as to the presence or absence of a gap on LGE MRI. However, these gaps detected by LGE MRI were not validated by invasive electrical assessment at the time of repeat catheter ablation.British Heart Foundation Clinical Research Training Fellowship."} +{"text": "Besides physical movements, yoga involves mental techniques which are considered to be essential for its effectiveness. However, active participation of the practitioners seems to be essential. This attitude can be operationalized and measured with the \u201cInner Congruence and Peaceful Harmony\u201d (ICPH) questionnaire. In a previous study, we have shown that ICPH can be predicted solely by the acceptance component of mindfulnes. Now we investigated whether and how variables of mental stability change with respect to ICPH.Prospective analysis of data from 160 individuals participating in a two-year yoga teacher training . Standardized questionnaires were administered at the start (t1), 3 months (t2), and 6 months (t3) later which comprised of Inner Congruence and Peaceful Harmony (ICPH), Mindfulness (FMI), Life Satisfaction (BMLSS), Positive Mood States (POMS/ASTS), health related Quality of Life (SF-12), Light-Heartedness / Easiness (LHE), and Aspects of Spirituality (ASP).During the course of intensified yoga practice, particularly LHE (Cohen\u00b4s d=.73) and mindfulness (d=.58) increased significantly , while positive mood (d=.29), mental health (d=.27) and ICPH (d=.22) increased only slightly. With respect to ASP, conscious interactions and religious orientation increased significantly (p<.01). Individuals with primarily low ICPH scores (28%) showed a significant development in mindfulness (d=.78) and LHE (d=.77), while those with moderate ICPH (53%) had a small increase of mindfulness (d=.47), but a strong increase in LHE (d=.93). Those with primarily high ICPH (19%) showed only small increases in mindfulness (d=.35) and LHE (d=.42).Even in already experienced yoga practitioners, positive mood and mindfulness increased significantly. One could suggest that ICPH represents a trait which may be developed to facilitate the beneficial effects of mindfulness on mental stability. Further investigations enrolling patients with chronic diseases are required."} +{"text": "The American Society of Extracorporeal Technology (AmSECT) has distributed a recommendation standard of practice and essential guidelines based on clinical evidence and currently accepted perfusion practice worldwide. These are integrated as learning objectives for clinical perfusion education. OnCloud\u00a9, an informatics and quality management program has been developed to collect and quantify procedural data to accurately produce quality control reports of perfusion practice incorporating these standards of practice.Fifty pediatric congenital heart surgery cases involving cardiopulmonary bypass (CPB) where a student was the primary perfusionist were retrospectively entered into the OnCloud\u00a9 database for quality control analysis. Thirty integrated OnCloud\u00a9 quality control indicators with predetermined threshold percentages were employed. When a threshold percentage was not achieved it was determined that further education was necessary to improve mastery of the learning objective correlating to the failed threshold of the quality indicator.The OnCloud\u00a9 quality control report reveled that six of the thirty quality indicators failed to meet the minimum thresholds. The report provided the indicator, the student achieved percentage, and the established threshold percentage. These included; pCO_2 35-45 mmhg (56.6%/85%), Hematocrit > 30% (86.5%/90%), Charting at 15 minute intervals (27.3%/99%), checklist completed (24.5%/99%), patient weaned from CPB (92%/99%), and procedure terminated without incident (94%/100%).Due to the dynamic learning environment perfusion education entails, course corrections must be undertaken to ensure mastery of critical aspects of the conduct of cardiopulmonary bypass. By using this software during clinical activity performed by the perfusion student, we can accurately assess deficiencies in clinical performance using evidence-based criteria and formulate a proper curriculum to ensure mastery."} +{"text": "In the VIKING study a 50 mg once daily regimen of the next generation HIV integrase inhibitor (INI) S/GSK1349572 (572) exhibited promising antiviral activity in an initial cohort (Cohort I) of treatment-experienced subjects with documented raltegravir (RAL) resistance. The purpose of this work was to evaluate via simulation techniques whether improved virologic responses could be achieved with higher doses in a second cohort (Cohort II) by utilizing pharmacokinetic (PK) and pharmacodynamic (PD) models derived using Cohort I data.10 VL (\u0394VL) at Day 11 was simulated for cohorts of 1000 subjects for each dose regimen according to different fixed levels of baseline fold change (FC) in 572 IC50 from wild type virus. The second set of simulations predicted responder percentages for clinical RAL-resistant HIV populations with diverse 572 susceptibilities. The data were well-described by the respective models. Model parameters were generally well-determined and VPC plots verified predictive performance.Interim PK data from VIKING Cohort I were combined with data from Phase 1 and 2a studies in healthy and HIV+, INI-na\u00efve subjects, respectively; data were analyzed using a linear two-compartment PK model. The population PK/PD analysis incorporated log10 HIV-1 RNA viral load (VL) sampled throughout 10 days of dosing during the Phase 2a and VIKING studies. The VL was modelled using an indirect response model in which the 572 plasma concentrations inhibited HIV-1 RNA production. Final PK and PK/PD models were validated using the visual predictive check (VPC) technique. Two sets of simulations were used to predict responder percentage for dose regimens proposed for Cohort II of VIKING. First, change from baseline in log10 \u0394VL at Day 11 by ~28%. Similarly, improvements in response of ~20% and ~18% were predicted for patient populations with HIV resistance profiles observed in RAL PhIIb and BENCHMRK virologic failure and VIKING screening populations, respectively. Our models predict 572 50mg twice daily will appreciably increase Day11 virologic responses in RAL-resistant subjects, supporting the dosing strategy for the ongoing Cohort II. 572 shows promise to demonstrate further the activity in this difficult to treat patient population.Simulations predicted increasing the dose regimen from 50 mg once daily to 50 mg twice daily would increase the percentage of patients with FC=8 that achieved \u2265 1.5 log"} +{"text": "Acinetobacter baumanii (AB) were identified in the Nashville General Hospital at Meharry epidemiology database for a three year period. Of these isolates, 77% were multi-drug resistant (MRAB). Mechanical ventilation and multiple site recovery were associated with MRAB, and MRAB isolates were associated with increased mortality relative to sensitive AB isolates [Two hundred and forty-seven isolates of isolates . AB acquAB MMC#4 was grown in plain LB broth or LB broth supplemented with MIC50 concentrations of levofloxacin, tobramycin, gentamicin, cefotaxime and meropenem. Cell pellets were lysed and total protein run on an SDS-PAGE gel. Protein bands were excised and in-gel digested with trypsin. Resulting peptides were analyzed using a Thermo Finnigan LTQ ion trap mass spectrometer equipped with a 1-D nanoLC pump (Eksigent), Nanospray source (James A Hill Company), and Xcalibur 2.0 SR2 instrument control (Thermo Scientific). Peptides were separated on a packed capillary tip with C18 reverse phase resin . Tandem spectra were acquired using a data dependent scanning mode in which one full MS scan (m/z 200-2000) was followed by 5 MS-MS scans. Tandem spectra were searched against the NCBI AB strain ACICU database using MryiMatch and IDPicker software. The search database was concatenated with the reverse sequences to determine false discovery rates. Proteins identified by less than two peptide spectral were eliminated (FDR <5%) and the output was filtered using IDPicker using a false positive ID threshold of 5%. Protein reassembly from identified peptide sequences was done as described by Zhang et al. [This analysis resulted in the identification of 125 high-confidence hits. Ten of these are presented in Table"} +{"text": "Dobutamine stress cardiovascular magnetic resonance (DCMR) is a robust tool for determining the presence of inducible ischaemia in patients with known or suspected coronary artery disease (CAD).To assess the safety of DCMR in clinical practice in a tertiary referral centre in the UK.We retrospectively studied all DCMR scans performed between May 2006 and March 2009.CMR studies were performed using a 1.5 T (Siemens Avanto) clinical CMR scanner. After a full functional assessment using steady state free precession (SSFP) cine imaging, three short axis images together with the three long axis images were selected and these were then acquired at each dose increment during a standard dobutamine-atropine protocol. Dobutamine was infused in 10 mcg/ kg/ min increments up to a maximum of 40 mcg/ kg/ min. Initial 5 mcg/ kg/ min increments were used where there was a wall motion abnormality at rest. The protocol was continued until target heart rate was achieved or until there was a recognised indication to stop prior to reaching target heart rate. Side effects and complications were recorded.Out of 455 patients, 21(4.6%) patients were unable to undergo the procedure 434 scans were performed in patients with a mean age of 64 years (range 13-86). 419 patients had a full dose study to assess for inducible ischaemia while 15 patients had a low dose protocol to determine myocardial viability.+ 9 micrograms/ kg/ min with additional atropine used in 119 (27.4%) patients. Target heart rate was achieved in 334 (79%) patients.The average dose of dobutamine required for each full dose study was 24 Of the 85 patients who failed to achieve target heart rate, 37(43%) developed significant chest pain, requiring the infusion to be stopped. Chest pain was the commonest cardiac side effect but in only 20% of these patients was the scan stopped early. 5 patients needed in-patient overnight observation for hypotension (2), broad complex tachycardia (1) and fast AF (2). None of these had a significant rise in troponin.Minor non-cardiac side effects occurred in 18 (4%) patients (eg nausea).Our experience suggests DCMR is a safe and feasible technique in routine clinical practice for assessing patients with suspected or known CAD and has a low complication rate."} +{"text": "Darunavir is one of the protease inhibitors that is recommended to treat protease inhibitor-na\u00efve or -experienced patients. Recent studies have determined the spectrum of darunavir activity in patients failing to antiretroviral therapy. Darunavir resistance mutations in protease gene have been identified and allow to classify viruses as sensitive (<3 mutations), possibly resistant (3 mutations) or resistant (\u22654 mutations) to darunavir.1583 genotypic resistance tests performed between 2008 and 2009 for darunavir-na\u00efve patients experiencing virological failure were analyzed retrospectively. Protease gene was sequenced and aminoacid changes analyzed at time of virological failure. The number of darunavir resistance mutations were determined and the strains were classified regarding the spectrum of darunavir activity .Among these experienced patients failing antiretroviral therapy, 63% harbored no darunavir mutations in the protease gene and 12%, 16%, 4% and 5% harbored 1, 2, 3 and at least 4 darunavir mutations, respectively. Patients with viruses harboring darunavir mutations had lower HIV-1 viral load than patients with viruses without any darunavir mutations.This study shows that the percentage of genotypic fully resistant strains to darunavir is rare in a population of darunavir-na\u00efve patients experiencing virological failure. A large proportion of patients harbored viruses without any darunavir resistance mutations allowing the use of darunavir/r (800/100 mg) QD. Virological failures without selection of any mutations showed higher viral load level rebound probably related to lack of adherence."} +{"text": "Cardiac magnetic resonance (CMR) imaging studies are increasingly being used as surrogate endpoints in clinical trials. There have been no previous studies investigating what level of training is required to provide an accurate and robust analysis of these parameters.20 CMR studies of acute coronary syndromes were included in this study. 6 observers with three levels of training conducted the image analysis: Observers 1+4: expert SCMR level 3 operators; Observers 2+5: trainees with SCMR level 2 experience; Observers 3+6: a cardiologist and a clinical trial coordinator with no previous CMR experience. The latter level underwent a 4hr tutorial on how to use the software on 10 practice cases. All 6 observers analysed 20 studies, and re-analysed 10 studies 24 hours after. Volumes and mass were analysed using semi-automated software , infarct size (IS) was manually planimetered using a 2SD threshold.Intra-observer variability was assessed using intraclass correlation coefficient (ICC), inter-observer variability was assessed using Bland Altman plots for agreement. Intra-observer variability was low for volumes, and was highest for mass measurements. . When compared with the most experienced observer, inter-observer variability was highest for IS and increased with decreasing level of experience (Table Operator experience highly influences the accuracy of calculating CMR parameters. When CMR LV volumes and infarct size are used as surrogate endpoints for clinical trials, a SCMR level 3 observer is the recommended operator to perform the analysis. Based on the results of our study, a SCMR level 2 operator will require additional experience, and a non-experienced operator significant additional training.NIHR Cardiovascular Biomedical Research Unit."} +{"text": "Background: Previous research indicates increased prostate cancer risk for pesticide applicators and pesticide manufacturing workers. Although underlying mechanisms are unknown, evidence suggests a role of oxidative DNA damage.Objectives: Because base excision repair (BER) is the predominant pathway involved in repairing oxidative damage, we evaluated interactions between 39 pesticides and 394 tag single-nucleotide polymorphisms (SNPs) for 31 BER genes among 776 prostate cancer cases and 1,444 male controls in a nested case\u2013control study of white Agricultural Health Study (AHS) pesticide applicators.p-values for interactions between three-level pesticide exposure variables (none/low/high) and SNPs (assuming a dominant model), and the false discovery rate (FDR) multiple comparison adjustment approach.Methods: We used likelihood ratio tests from logistic regression models to determine NEIL3 [nei endonuclease VIII-like 3 (Escherichia coli)], which encodes a glycosylase that can initiate BER, was the most significant overall . Fonofos exposure was associated with a monotonic increase in prostate cancer risk among men with CT/TT genotypes for rs1983132 , whereas fonofos was not associated with prostate cancer risk among men with the CC genotype. Carbofuran and S-ethyl dipropylthiocarbamate (EPTC) interacted similarly with rs1983132; however, these interactions did not meet an FDR < 0.2.Results: The interaction between fonofos and rs1983132 in Conclusions: Our significant finding regarding fonofos is consistent with previous AHS findings of increased prostate cancer risk with fonofos exposure among those with a family history of prostate cancer. Although requiring replication, our findings suggest a role of BER genetic variation in pesticide-associated prostate cancer risk. Previous research has demonstrated significantly increased prostate cancer risk for pesticide applicators and pesticide manufacturing workers compared with the general population , suggestAccumulating DNA damage due to chronic oxidative stress has been proposed as an important mechanism in prostate carcinogenesis . PreviouGiven the potential importance of oxidative damage in pesticide-associated prostate cancer risk and the role of the BER pathway in repairing this type of damage, we conducted a nested case\u2013control study of white male pesticide applicators within the Agricultural Health Study (AHS) to evaluate interactions between pesticide exposures and genetic variation in 31 BER genes with respect to prostate cancer. We hypothesized that BER gene variants may modify pesticide-associated prostate cancer risk.Study population. The AHS prostate cancer nested case\u2013control study has been described in detail previously (a) were diagnosed with prostate cancer between 1993 and 2004 after enrollment in the AHS cohort, b) provided a buccal cell sample, and c) had no previous history of cancer except nonmelanoma skin cancer. Eligible controls were white male applicators in the cohort who a) provided a buccal cell sample, b) had no previous history of cancer except nonmelanoma skin cancer, and c) were alive at the time of case diagnosis. Previous work in the AHS has demonstrated minimal differences with respect to a variety of characteristics between participants that did and did not provide a buccal cell sample (n = 20) or < 90% completion rate for genotyping assays (n = 88)], or a genetic background that was inconsistent with European ancestry resulted in a final sample size of 776 cases and 1,444 controls. All participants provided written informed consent, and the study was approved by the institutional review boards of all participating institutions.eviously . Brieflyl sample . Controll sample , exclusi (n = 3) or signiExposure assessment. Information on lifetime use of 50 pesticides was captured in two self-administered questionnaires completed during cohort enrollment (1993\u20131997). All 2,220 nested case\u2013control study participants completed the first (enrollment) questionnaire, which inquired about ever/never use of the 50 pesticides, as well as duration (years) and frequency (average days per year) of use for a subset of 22 of the pesticides; 1,439 of these men (60.4% of cases and 67.2% of controls) completed the second (take-home) questionnaire, which inquired about use of the remaining 28 pesticides. A previous AHS analysis demonstrated similar characteristics, except for age, between cohort participants who completed the take-home questionnaire and those who did not ( did not . For eac did not that wasGenotyping and single-nucleotide polymorphism (SNP) selection. DNA was extracted from buccal cells using the Autopure protocol . Genotyping was performed at the NCI Core Genotyping Facility using a custom Infinium\u00ae BeadChip assay (iSelect\u2122) from Illumina Inc. as part of an array of 26,512 SNPs in 1,291 candidate genes. Blinded duplicate samples (2%) were included, and SNP concordance ranged from 96% to 100%. Tag SNPs were chosen to cover candidate DNA repair genes for three ancestry populations in the HapMap Project . Two SNPs had ptrend < 0.01: rs3786662 (ptrend = 0.007), tagged for PNKP (polynucleotide kinase 3\u00b4-phosphatase), and rs246079 (ptrend = 0.008), tagged for the uracil-DNA glycosylase gene UNG.We identified 22 SNPs in 11 genes with NEIL3 [nei endonuclease VIII-like-3 (Escherichia coli)], DUT (deoxyuridine triphosphatase), POLB , and NTHL1 [nth endonuclease III-like-1 (E. coli)], met the FDR < 0.2 criterion (DUT (r2 = 0.61\u20131.00), one fonofos \u00d7 NEIL3 SNP interaction, and one glyphosate \u00d7 POLB SNP interaction]. However, 13 of the 14 combinations were qualitative interactions with a positive association with pesticide exposure among men in one genotype group and an inverse association for men in the other genotype group. The exception was fonofos \u00d7 NEIL3 rs1983132. There was a significant monotonic increase in prostate cancer risk with increasing fonofos exposure among men with CT/TT genotypes for rs1983132 , but no association among men with the CC genotype for a summary of all interactions evaluated]. We observed a similar pattern of interaction for fonofos with the moderately correlated NEIL3 SNP rs17064578 (r2 = 0.32), although this finding did not meet FDR < 0.2 . Rs1983132 showed low correlations with other NEIL3 SNPs (r2 \u2264 0.15), and analysis of interactions between NEIL3 haplotypes and fonofos also suggested that rs1983132 might be driving our fonofos \u00d7 NEIL3 SNP interaction findings. We observed borderline significant or significant interactions between fonofos and three of four haplotypes that included the variant T allele for rs1983132, including one without the variant C allele for rs17064578, but we did not observe evidence of an interaction with a haplotype that contained the variant allele for rs17064578 and the C allele for rs1983132 such that each pesticide was associated with prostate cancer among men with CT/TT genotypes for this locus , whereas neither pesticide was associated with prostate cancer among men with the CC genotype , TDG , LIG1 , and POLE but did not estimate a significant interaction with the haplotype that contained the variant allele for rs812498 and the wild-type allele for rs322107 . When we included the five terbufos \u00d7 LIG1 SNP interactions in a single model, terbufos \u00d7 LIG1 rs3786763 remained borderline significant (pinteract = 0.06). Neither the XRCC1 SNPs nor the POLE SNPs could be modeled together because of their high correlations.Other interactions that met the criteria described above included interactions between fonofos, terbufos, and atrazine and correlated SNPs within = 0.88] .When we p < 0.01 for both intensity-weighted and unweighted pesticide exposure metrics, and a significant monotonic increase in prostate cancer risk with increasing exposure in one genotype group and no significant association in the other and EPTC (a thiocarbamate herbicide) showed similar patterns of interaction with rs1983132, although these interactions were weaker and did not remain significant after adjustment for multiple comparisons. The risk of prostate cancer associated with exposure to fonofos, carbofuran, or EPTC alone among men with CT/TT genotypes for rs1983132 appeared to be increased for those exposed to two or more of these pesticides. However, because of relatively wide and overlapping CIs, it is unclear whether the joint effect of these pesticides was driven by fonofos alone.Fonofos (an OP insecticide) interacted similarly with two moderately correlated NEIL3 rs1983132 interaction finding, in vitro, experimental animal, and human biomonitoring studies suggest that some OP insecticides might induce oxidative stress and related DNA damage with exposure to carbofuran or products of its nitrosation , which is not part of the BER pathway, and rs246079, located in an intronic region of UNG but also tagged for ALKBH2 , which is involved in the direct reversal of DNA damage but not BER.We did not observe highly significant BER SNP main effects in our study. Only two SNPs had a XRCC1 R399Q (rs25487), PARP1 [poly (ADP-ribose) polymerase 1] V762A (rs1136410), or OGG1 (8-oxoguanine DNA glycosylase) S326C (rs1052133), although some previous studies have observed phenotypic changes and altered prostate cancer susceptibility with genetic variation at these loci PDFClick here for additional data file."} +{"text": "Inducing arteriogenesis using stem cells is a promising treatment for patients suffering from peripheral arterial disease (PAD). Computed tomography angiography (CTA) is often used as a gold standard method for evaluating vessel diameters in ischemic tissue after therapy.The purpose of our study was to compare the vessel area measurement in Time-of-Flight (TOF) MR angiography (MRA), contrast-enhanced MRA (ceMRA) and CTA after transplantation of mesenchymal stem cells (MSCs) in an animal PAD model.3 voxel size), 3D ceMRA acquired in the coronal plane and a C-arm CTA were acquired in random order prior to SFA occlusion, and 7 and 14 days after occlusion. Vessel areas at seven locations were measured , empty capsules (n=4) or microencapsulated MSCs (n=5) in the medial thigh 24h after endovascular occlusion of the left superficial femoral artery (SFA). Three-dimensional (3D) TOF MRA acquired in the axial plane images.Vessel areas in the aorta were 0.116 \u00b1 0.02cmMeasurement of small vessel diameters by MRA is challenging especially in ischemic or atherosclerotic disease. TOF MRA is traditionally considered a poor method for measuring vessel size in areas of slow flow. However, agreement of TOF MRA with CTA was higher than ceMRA in our rabbit PAD model. Potential degradation of ceMRA image resolution occurred due to reformatting of the ceMRA in the axial plane for measurement. However, both techniques showed an acceptable precision to measure small vessel diameters without ionizing radiation for the assessment of efficacy of an arteriogenesis therapy."} +{"text": "Perfusion-cardiac magnetic resonance (CMR) is generally accepted to assess myocardial ischemia non-invasively by assessing the ischemia noninvasively with moderate coronary artery stenosis and follow up of these patients will helps in prognostic estimation.Retrospective study of 68 patients with moderate coronary artery stenosis (50-70%) underwent stress and viability CMR in the period from January 2010 to December 2011. Patients are referred by cardiologists and physicians after coronary angiogram for further evaluation.Cardiac MR done with state-of-the-art MR equipment Magnetom Avanto 1.5 T (Siemens) by using stress and viability sequences.Follow up of these patients are done for major adverse cardiac events -MACE for 9 months.Of 68 patients with CAD, 94 segments of moderate coronary artery stenosis are found. Stress CMR shows evidence of ischemia in 54 segments & infarcts in 4 patients.Follow up these patients for major adverse cardiac events in 9 months , 18 patients had MI , 6 patients had unstable angina and 5 patients with LV dysfunctionStress CMR analyses patients with moderate coronary artery stenosis for ischemia and predicts the development of major adverse cardiac eventsNo funding taken."} +{"text": "Implementation of assisted mechanical ventilation in acute lung injury (ALI) patients may decrease ventilator-induced lung injury by redistribution of tidal ventilation towards dependent lung regions. Up to now, tidal ventilation regional distribution has been measured by expensive and complicated methods, not readily available at the bedside. Electrical impedance tomography (EIT) is a relatively new non-invasive bedside method to monitor tidal ventilation distribution, validated in preclinical studies. We verified the feasibility of using EIT to monitor tidal ventilation regional distribution in patients undergoing assisted ventilation and we describe the effect of different pressure support levels on regional ventilation.\u00ae; Dr\u00e4ger Medical GmbH, L\u00fcbeck, Germany), dividing the lung imaging field into four contiguous same-size regions of interest (ROI): ventral right (ROI 1) and left (ROI 2), dorsal right (ROI 3) and left (ROI 4). We randomly performed two steps of PS ventilation for 15 minutes, leaving the positive end-expiratory pressure (PEEP) and FiO2 unchanged: PSlow (p0.1 \u22652 cmH2O) and PShigh (p0.1 <2 cmH2O). At the end of each step, we recorded: ventilation parameters, arterial blood gas analysis and percentage of tidal ventilation distribution in the four ROIs. Analyses were performed by paired t test.We enrolled 11 consecutive ALI patients admitted to our ICU, intubated and undergoing pressure support (PS) ventilation. We monitored the regional tidal ventilation distribution by means of a new EIT monitor and peak inspiratory pressure . At PShigh, proportional distribution of tidal ventilation significantly changed in all four ROIs , moving from dorsal to ventral.The ALI etiology was: trauma (18%), septic shock (18%), pneumonia (46%) and postoperative respiratory failure (18%). PSEIT may be a useful tool to monitor lung regional ventilation at the bedside. PS levels that blunt patient effort may promote redistribution of tidal ventilation towards ventral lung regions."} +{"text": "A beneficial effect of acetylsalicylic acid (ASA) on vein graft patency has been described, but some patients experience adverse cardiac events despite appropriate ASA treatment. Study aim was to define ASA resistance using Multiple electrode aggregometry (MEA) preoperatively in group of patients undergoing coronary artery bypass grafting (CABG).Prospective observational trial at University Hospital Center Zagreb enrolled 131 patients scheduled for CABG, and divided them into 4 groups with respect to preoperative antiplatelet therapy (APT). Group 1 received 100 mg ASA per day, Group 2 100 mg ASA + 75 mg clopidogrel per day, Group 3 75 mg clopidogrel per day, and Group 4 did not receive any APT. MEA with ASPI test (sensitive to ASA) and ADP test (sensitive to clopidogrel) was performed prior to surgery. In Group 1, patients were characterized as ASA resistant if their ASPI test value exceeded the 75th percentile distribution.Study enrolled 131 patients. Significant differences both in the ASPI (p<0.001) and the ADP test (p=0.038) were observed between patients in different APT groups. In Group (1) ASPI test value of 30 AUC presented 75th percentile of distribution, thus indicating ASA resistance. Group 2 patients had slightly lower ADP test values, but no significant difference occurred . In Group 1 and 2, significant correlations between the ADP test and both, platelet count and fibrinogen level were observed.Association between low response to ASA and post-CABG major adverse ischemic events risk increase has been described thus indicating need for ASA resistant patients detection using bedside suitable and drug specific platelet function tests. In patients with preoperative ASPI test exceeding 30 AUC postoperative, ASA dose adjustment or clopidogrel addition according to MEA results should be considered."} +{"text": "The chromatin remodeling protein ATRX, which targets tandem repetitive DNA, has been shown to be required for expression of the alpha globin genes, for proliferation of a variety of cellular progenitors, for chromosome congression and for the maintenance of telomeres. Mutations in ATRX have recently been identified in tumours which maintain their telomeres by a telomerase independent pathway involving homologous recombination thought to be triggered by DNA damage. It is as yet unknown whether there is a central underlying mechanism associated with ATRX dysfunction which can explain the numerous cellular phenomena observed. There is, however, growing evidence for its role in the replication of various repetitive DNA templates which are thought to have a propensity to form secondary structures. Using a mouse knockout model we demonstrate that ATRX plays a direct role in facilitating DNA replication. Ablation of ATRX alone, although leading to a DNA damage response at telomeres, is not sufficient to trigger the alternative lengthening of telomere pathway in mouse embryonic stem cells. ATRX is an SNF2-related chromatin remodelling protein which acts with the histone chaperone DAXX to insert the variant histone H3.3 into telomeric and pericentromeric heterochromatin in vitro. In the absence of ATRX there is an increase in DNA damage at telomeres An important recent finding is that ATRX localises to G-rich tandem repeats including interstitial repeats and telomeres during S phase Here we confirm and extend these important observations in a primary mouse cell line knocked out for ATRX and demonstrate a defect in S phase progression following an aphidicolin block, sensitivity to hydroxyurea, increased stalling of replication and an increase in double strand breaks (DSBs) as detected by 53BP1 foci and a neutral comet assay. These aspects of ATRX function may be particularly important at telomeres. It has previously been shown that, in telomerase negative tumour cells, viability is maintained via an alternative pathway (ALT) to lengthen telomeres. Furthermore, tumours with the ALT phenotype frequently harbour acquired somatic mutations in the ATRX gene nullAtrx) with their wildtype counterparts (Atrxflox) mES cells Atrxnull cells upon treatment with the DNA replication stress inducing drug hydroxyurea (HU) but no response to other DNA replication inhibitors (camptothecin and aphidicolin) or DNA damaging agents (gamma-irradiation (IR) and cisplatin) cell lines conditionally deleted for full length ATRX (splatin) . Whilst Atrxflox and Atrxnull mES cells were blocked at the G1/S boundary with aphidicolin, released, and the progression through S-phase was monitored by fluorescence activated cell sorting (FACS) showing that S-phase was markedly prolonged in the Atrxnull cells and chlorodeoxyuridine (CldU)), lysed, the DNA fibres spread on glass slides, probed with specific antibodies and visualised by fluorescence microscopy reatment , the dison forks [17], [1ox cells . After rll cells . A smallll cells but thisll cells . No consl clones , suggestAtrxnull cells relative to the Atrxflox cells using both quantitative FISH, which gave median telomere intensities of 1020 (A.U.) for both Atrxflox and Atrxnull cells , ATRX interaction partners were immune-isolated from a HeLa nuclear extract and resolved by SDS PAGE gel electrophoresis. Three major bands were analysed by mass spectrometry . In addiImmunoblotting with RAD50, MRE11 and NBS1 specific antibodies confirmed the identification of these proteins as ATRX interaction partners . These iThe MRN complex has many known functions including: double strand break repair (via both HR and NHEJ) and the restart of stalled replication forks Here we show that a conditional deletion of full length ATRX in primary mouse ES cells undergoing replicative stress, leads to a prolongation in S-phase, concomitant accumulation of gamma-H2AX and an increase in fork stalling. Notably, loss of ATRX elicits an apparent defect in late S phase, which likely includes replication of repetitive, heterochromatic sites, known targets of ATRX binding This model has gained support from a number of complementary studies. Ablation of ATRX in mouse myoblasts delayed their progression in S phase and led to an accumulation of DNA damage including telomere fragility nullAtrx cells to hydroxyurea, a replication inhibitor that increases the likelihood of G4 formation. Moreover, in line with these observations, nullAtrx neuroprogenitor cells have recently been shown to be sensitive to the G-quadruplex stabilising ligand telomestatin A large proportion of ATRX target sites are predicted to adopt non-B form secondary structures, including the G-quadruplex conformation nullAtrx cells, thereby suggesting that loss of ATRX alone is not sufficient to trigger ALT in this cellular context. It is therefore obviously of interest to determine the additional requirements for ALT activation.Interestingly, a role for ATRX in facilitating replication through potential G-quadruplex forming sequences may shed light on its recently ascribed role as a tumor suppressor in a specific subset of malignancies that depend on a telomerase-independent pathway of telomere maintenance called the \u2018alternative lengthening of telomeres\u2019 (ALT) pathway By co-immunoprecipitation we demonstrate an interaction between endogenous ATRX and components of the MRE11-RAD50-NBS1 (MRN) complex during S phase and a similar finding, using an expressed tagged ATRX transgene, has recently been reportedCell cycle synchronisation was performed using a thymidine aphidicolin double block as previously described mES cells were treated with ionising radiation at the dosages specified and the Comet assay was performed as described in Immunoprecipitations were performed from 2 to 4 mg HeLa nuclear extract (Cilbiotech) in IP buffer . Immunoprecipitations were performed overnight at 4\u00b0C using 3 \u03bcg anti-ATRX antibody (sc-15408) and 4 \u03bcg anti-RAD50 antibody (Abcam ab89) with Protein A or G agarose beads in the presence or absence of 100 \u03bcg/ml ethidium bromide. Beads were washed 4 times in IP buffer and bound proteins eluted into SDS loading buffer (Laemmli) by heating at 90\u00b0C for 5 mins. Western blotting was performed using the following additional antibodies; anti-MRE11 (Abcam ab214), anti-NBS1 (Santa Cruz sc-11431).Telomere length was determined by terminal restriction length analysis Cells grown on coverslips were prepared for IF by standard procedures. The following antibodies were used for immunostaining: anti-ATRX (Santa Cruz sc-15408); anti-ATRX 39f Image analysis was done using custom perl processing scripts that called various ImageJ macros to allow filtering and automatic thresholding methods to segment nuclei and then identify foci per nucleus. The JACoP This assay was performed according to a previously published method 150-300 mES cells were grown for 24h before treatment with the specified DNA damaging agents or replication inhibitors. After 3 or 4 days, colonies were stained with 1% w/v methylene blue 50% ethanol and counted..For fibre analysis mES cells were incubated with 25 \u03bcM IdU for 10 mins or 90 mins in the presence of 1 mM hydroxyurea, followed by a 10 or 20 min incubation with 250 \u03bcM CldU respectively. For assessment of fork processivity mES cells were incubated with IdU for 10 mins followed by a 40 min incubation with CldU in the presence of 1 mM hydroxyurea. Spreading and visualisation of fibres was performed as previously described Figure S1floxAtrx and nullAtrx mES cells following 8 hours release from aphidicolin block. Black box shows late replicating population. (B) Percentage of late replicating cells as determined by FACs following 2, 6 and 8 hours post release from aphidicolin block. Error bars indicate \u00b1 SEM from three independent experiments (C) Immunoblot and quantitation (D) to assess levels of gamma-H2AX in histones purified from floxAtrx and nullAtrx mES cells (clone 2) at the indicated time points. This showed an elevated DDR, reflected by elevated gamma-H2AX, in nullAtrx cells as compared to the floxAtrx cells. U \u200a=\u200a unsynchronised cells. Histone H3 is shown as the loading control. Error bars indicate \u00b1 SEM. (E) Results of fibre analysis showing relative frequencies of replication intermediates in in floxAtrx and nullAtrx mES cells (clone 2) with hydroxyurea treatment during the IdU pulse.(A) Representative images of cell cycle profile for (TIF)Click here for additional data file.Figure S2Representative images for immunofluorescence in wildtype mES cells showing 3 way co-localisation between PCNA, MRE11 and ATRX using an N-terminal specific ATRX antibody. Nuclei are outline with a dashed white line.(TIF)Click here for additional data file.Figure S3Unwinding assay for G4 DNA. Recombinant ATRX (TIF)Click here for additional data file."} +{"text": "Haemophilus influenzae [NTHi]) infection. However, the mechanisms underlying bacterial infections during AECOPD remain poorly understood. As neutrophilic inflammation including increased release of human neutrophil elastase (HNE) is a salient feature of AECOPD, we hypothesized that HNE impairs airway epithelial defense against NTHi by degrading airway epithelial host defense proteins such as short palate, lung, and nasal epithelium clone 1 (SPLUNC1).Acute exacerbations of chronic obstructive pulmonary disease (AECOPD) are a significant cause of mortality of COPD patients, and pose a huge burden on healthcare. One of the major causes of AECOPD is airway bacterial HNE directly increased NTHi load in human airway epithelial cells; (2) HNE degraded human SPLUNC1 protein; (3) Recombinant SPLUNC1 protein reduced NTHi levels in HNE-treated human airway epithelial cells; (4) NTHi levels in lungs of SPLUNC1 knockout mice were increased compared to wild-type mice; and (5) SPLUNC1 was reduced in lungs of COPD patients.Recombinant human SPLUNC1 protein was incubated with HNE to confirm SPLUNC1 degradation by HNE. To determine if HNE-mediated impairment of host defense against NTHi was SPLUNC1-dependent, SPLUNC1 protein was added to HNE-treated primary normal human airway epithelial cells. The Our findings suggest that SPLUNC1 degradation by neutrophil elastase may increase airway susceptibility to bacterial infections. SPLUNC1 therapy likely attenuates bacterial infections during AECOPD. One of the prominent inflammatory cells in COPD lungs is neutrophils Because airway epithelial cells represent the first line of lung responses to invading bacteria, we sought to determine if HNE impairs airway epithelial host defense functions. In the current study, we focused on the effects of HNE on the function of a recently described host defense protein short palate, lung, and nasal epithelium clone 1 (SPLUNC1). A previous study suggests that HNE is involved in SPLUNC1 degradation Mycoplasma pneumoniae (Mp) \u2212/\u2212 mice, we further confirmed the in vivo host defense function of SPLUNC1 during Mp infection SPLUNC1 is produced by large airway epithelial cells in humans and mice We hypothesized that HNE impairs human lung defense functions by decreasing airway epithelial SPLUNC1 levels. Our current study demonstrates that: (1) HNE directly reduced human airway epithelial defense against NTHi; (2) HNE degraded human SPLUNC1 protein; (3) Recombinant SPLUNC1 protein reduced NTHi levels in HNE-treated human airway epithelial cells; (4) NTHi levels in lungs of SPLUNC1 knockout mice were increased compared to wild-type mice; and (5) SPLUNC1 was reduced in lungs of COPD patients. Together, our findings provide a novel mechanism underlying bacterial infections in COPD patients, and a potential therapy to more effectively treat AECOPD.http://www.iiam.org).Experimental animals used in this study were covered by a protocol approved by Institutional Animal Care and Use Committee (IACUC) of National Jewish Health, Denver, Colorado, USA. All experimental procedures were carried out to minimize animal discomfort, distress, and pain by following the American Veterinary Medical Association Guidelines. All human materials such as bronchoalveolar lavage used in this study were approved by Institutional Review Board (IRB) of National Jewish Health, Denver, Colorado, USA. The written informed consent was waived by National Jewish Health IRB for de-identified organ donors without lung diseases from whom primary normal human tracheobronchial epithelial cells were obtained through the International Institute for the Advancement of Medicine (IIAM) , and diluted in 0.05 M sodium acetate solution with 0.1 M NaCl (pH 5.0). SPLUNC1 protein at 10 \u00b5g/ml, a physiologic dose as we reported Primary normal human tracheobronchial epithelial cells were obtained from tracheas and bronchi of deidentified organ donors by digesting the tissue with 0.2% protease solution and then subjected to air-liquid interface (ALI) cultures, as we reported Haemophilus influenzae (NTHi) strain 12 was plated on chocolate agar plates and incubated overnight at 37\u00b0C in 5% CO2. A single colony was used to inoculate 10 ml of brain heart infusion broth , and allowed to grow overnight. The subculture was then washed twice with PBS and the optical density (O.D.) at a wavelength of 620 nm was determined to obtain the colony-forming unit (CFU) Nontypeable 3 CFU/transwell To determine if HNE treatment impairs epithelial defense against NTHi, cells on day 10 of ALI culture were treated at the apical surface with 50 \u00b5l of HNE (10 \u00b5g/ml) in the presence or absence of NTHi at 10To test if SPLUNC1 reduction in HNE-treated cells is responsible for increased NTHi levels, recombinant human SPLUNC1 protein (10 \u00b5g/ml) or control protein from HEK293 cells was added to the apical surface of epithelial cells after 2 hrs of HNE treatment and NTHi infection. After 46 hrs of SPLUNC1 treatment or after 48 hrs of NTHi infection, apical supernatants and cells were harvested for NTHi quantification.in vivo function of SPLUNC1 against NTHi. SPLUNC1 KO mice on the C57BL/6 background were generated in our laboratory and bred under pathogen-free housing conditions as previously reported 5 colony forming unit [CFU]/mouse in 50 \u00b5l saline), and sacrificed after 24 hrs of infection to obtain the lung tissue for NTHi quantification. Briefly, lung tissues were homogenized and plated on chocolate agar paltes to count NTHi CFUs. All the experimental protocols were approved by Institutional Animal Care and Use Committee (IACUC) at National Jewish Health.SPLUNC1 knockout (KO) mouse model was used to reveal the We obtained bronchoalveolar lavage (BAL) fluid from healthy non-smokers and smokers, as well as smokers with moderate to severe COPD patients to measuP<0.05 was considered significant.One-way analysis of variance (ANOVA) was used for multiple comparisons, and a Turkey\u2019s post hoc test was applied where appropriate. To test if HNE directly degrades SPLUNC1, recombinant human SPLUNC1 protein at a physiological dose (10 \u00b5g/ml) was incubated with HNE at various doses for 30 minutes. Western blot analysis demonstrated that HNE treatment significantly reduced intact SPLUNC1 (25-kD), and resulted in the formation of a 22-kD SPLUNC1 fragment (loss of a 3-kD peptide) . We thenHaving shown the ability of HNE to degrade recombinant SPLUNC1 protein in a cell-free environment, we tested if HNE reduces SPLUNC1 protein secreted from primary normal human airway epithelial cells that were well differentiated under air-liquid interface culture. HNE dose-dependently decreased SPLUNC1 after 4 hrs of HNE treatment . HNE effTo determine the effects of HNE on bacterial load, apical surface of cultured well-differentiated normal human airway epithelial cells was infected with NTHi for 48 hrs in the presence or absence of HNE. NTHi levels in the apical supernatants and cellTo determine if HNE-mediated SPLUNC1 reduction is responsible for impaired airway epithelial defense against NTHi , recombinant human SPLUNC1 protein (10 \u00b5g/ml) was added 2 hrs after HNE treatment and NTHi infection. After 48 hrs of infection, NTHi levels were measured in apical supernatants and cells. First, in line with data shown in in vivo function of SPLUNC1 during NTHi infection is unclear. We utilized our SPLUNC1 knockout (SPLUNC1\u2212/\u2212) mouse model to delineate the in vivo function of SPLUNC1. After 24 hrs of an intranasal inoculation of NTHi, bacterial load in SPLUNC1\u2212/\u2212 mice was about 30-fold higher than that in wild-type mice -mediated SPLUNC1 degradation impairs airway epithelial defense against bacterial infection. We found that HNE markedly reduces SPLUNC1 in well-differentiated human airway epithelial cells and subsequently increases bacterial load, which can be attenuated by recombinant human SPLUNC1 protein. Moreover, we found decreased SPLUNC1 in human COPD lungs, and increased lung bacterial load in SPLUNC1 knockout mice. Together, our data suggest that increased HNE, a feature of excessive inflammation during acute exacerbations of COPD and other chronic lung diseases, may contribute to bacterial infections in part through degrading host defense protein SPLUNC1.Pseudomonas aeruginosa) infection Acute exacerbations of COPD (AECOPD) pose the highest risk to patients and the greatest costs for COPD-related healthcare. Respiratory bacterial and viral infections are the major cause of AECOPD, which commonly recruit neutrophils into the airway lumen and activate them. During neutrophil activation, they release various mediators including HNE. Although the NE knockout mouse model suggests a beneficial role of intracellular NE in host defense against bacterial . Therefore, SPLUNC1 reduction in stable COPD patients may be caused by other factors such as cigarette smoke exposure. In a pilot study, we exposed well-differentiated human airway epithelial cells to whole cigarette smoke or air (control) for 10 minutes as we described before Although the current study was focused on Pseudomonas aeruginosa. Third, HNE may degrade other antimicrobial substances studies are warranted to determine the physiological significance of a less than one log bacterial load change in cultured well-differentiated human primary airway epithelial cells treated with recombinant SPLUNC1 protein and/or HNE.Our current study has several limitations. First, we only used normal human airway epithelial cells to study the impact of HNE on SPLUNC1 degradation and bacterial infection. We will consider COPD airway epithelial cells in our future studies to determine if COPD cells may differ from normal cells in their responses to HNE and NTHi infection. Second, since NTHi was the only strain of bacteria examined in the current study, our results should be verified in other strains of bacteria involved in COPD, including In summary, our current study provides a new mechanism to explain increased susceptibility of COPD patients to bacterial infections, in particular during acute exacerbations that are featured with excessive neutrophilic inflammation and release of neutrophil elastase. As SPLUNC1 is an endogenous protein and unlikely has the side effects of antibiotics, it has the potential to treat bacterial infections associated with AECOPD and perhaps other lung diseases such as asthma and cystic fibrosis."} +{"text": "Eucalyptus EgMYB2 and the pine PtMYB4, were shown to differentially bind to and activate the eight variants of the 7-bp SMRE consensus sequence, composed of ACC(A/T)A(A/C)(T/C). Together, our results indicate that the tree MYBs, PtrMYB2/3/20/21, EgMYB2 and PtMYB4, are master transcriptional switches that activate the SMRE sites in the promoters of target genes and thereby regulate secondary wall biosynthesis during wood formation.Wood is mainly composed of secondary walls, which constitute the most abundant stored carbon produced by vascular plants. Understanding the molecular mechanisms controlling secondary wall deposition during wood formation is not only an important issue in plant biology but also critical for providing molecular tools to custom-design wood composition suited for diverse end uses. Past molecular and genetic studies have revealed a transcriptional network encompassing a group of wood-associated NAC and MYB transcription factors that are involved in the regulation of the secondary wall biosynthetic program during wood formation in poplar trees. Here, we report the functional characterization of poplar orthologs of MYB46 and MYB83 that are known to be master switches of secondary wall biosynthesis in Arabidopsis. In addition to the two previously-described PtrMYB3 and PtrMYB20, two other MYBs, PtrMYB2 and PtrMYB21, were shown to be MYB46/MYB83 orthologs by complementation and overexpression studies in Arabidopsis. The functional roles of these PtrMYBs in regulating secondary wall biosynthesis were further demonstrated in transgenic poplar plants showing an ectopic deposition of secondary walls in PtrMYB overexpressors and a reduction of secondary wall thickening in their dominant repressors. Furthermore, PtrMYB2/3/20/21 together with two other tree MYBs, the Wood is produced by the activity of the vascular cambium, and it encompasses a complex developmental program involving the differentiation of the vascular cambium into secondary xylem mother cells, cell elongation, secondary wall deposition, programmed cell death, and finally heartwood formation Brachypodium, and alfalfa Wood at maturity is essentially the remains of secondary walls largely composed of cellulose, hemicelluloses and lignin. Therefore, understanding how the biosynthesis of secondary wall components is regulated could potentially provide genetic tools for altering wood composition. Since many genes are required for the biosynthesis of each major wood component, it is conceivable that the biosynthetic genes responsible for the making of wood components are coordinately activated during wood development. Recent molecular and genetic studies in tree species have demonstrated that the coordinated activation of wood biosynthetic genes is mediated by a transcriptional network involving multileveled transcriptional controls. It was found that a group of wood-associated NAC domain transcription factors (WNDs) are the top master switches regulating the expression of a number of downstream transcription factors, which ultimately lead to the biosynthesis of secondary walls during wood formation in tree species PtrMYB3 and PtrMYB20, together with two other MYB genes, EgMYB2PtMYB4Eucalyptus and pine, respectively, have previously been shown to be functional orthologs of the Arabidopsis MYB46 and MYB83The poplar MYB genes, In this report, we investigated the effects of overexpression and dominant repression of poplar orthologs of MYB46 and MYB83 on wood formation in poplar trees, and determined the cis-elements these tree MYBs bind. We show that beside the previously described PtrMYB3 and PtrMYB20, two additional MYBs, PtrMYB2 and PtrMYB21, are functional orthologs of MYB46 and MYB83, capable of activating the biosynthetic genes for cellulose, xylan and lignin. Overexpression and dominant repression of these PtrMYB in transgenic poplar lead to an ectopic deposition of secondary walls and a reduction in secondary wall thickening, respectively. Furthermore, we demonstrate that the tree MYBs, including PtrMYB2/3/20/21, EgMYB2 and PtMYB4, all bind to and activate the SMRE sequences. Our findings indicate that MYB46 and its orthologs in both herbaceous Arabidopsis and tree species activate their downstream target genes via binding and activating the SMRE sites.PtrMYB2 and PtrMYB21 is induced by PtrWNDs, indicating that PtrMYB2 and PtrMYB21 are PtrWND-regulated downstream transcription factors involved in transcriptional regulation of wood formation PtrMYB2 and PtrMYB21 are grouped together with the Arabidopsis MYB46 and MYB83 genes, two other poplar MYB genes (PtrMYB3 and PtrMYB20) that were previously shown to be functional orthologs of MYB46 myb46 myb83 double mutant. The myb46 myb83 mutant exhibited a strongly retarded seedling growth phenotype and a defect in secondary wall thickening in leaf vessels . Because PtrMYB3 and PtrMYB20 are duplicated genes and so are PtrMYB2 and PtrMYB21PtrMYB3 and PtrMYB21, one from each pair of the duplicated genes for further analysis. Transgenic poplar MYB overexpressors had shorter stems with smaller leaves compared with the control ones transformed with the empty vector. Staining of stem sections for lignin, xylan and cellulose revealed that in addition to the normal staining of secondary wall components in secondary xylem and phloem fiber cells as seen in the control expressing the dominant repressors showed a reduced stem height and strong alterations in wall thickness and vessel morphology in the wood of stems and PtrMYB21 (5\u2032-atgaggaagccagaggcctct-3\u2032 and 5\u2032-tcattggaaatcaaggaatggaaaggc-3\u2032) driven by the 3-kb MYB46 promoter were cloned into the pGPTV vector and introduced into the myb46 myb83 double mutant myb46 (\u2212/\u2212) myb83 (\u2212/\u2212) mutants by PCR amplification of T-DNA insertions as described previously myb46 myb83 double mutant plants that showed normal growth as the wild type were chosen for examination of plant growth and vessel morphology phenotypes.The full-length cDNAs of PtrMYB2, PtrMYB21, or PtrMYB3 (5\u2032-atgaggaagccggatctaatg-3\u2032 and 5\u2032-tgtaagtgtagcctgttataaaacttgg-3\u2032) cDNA downstream of the CaMV 35S promoter in pBI121. The PtrMYB dominant repression constructs (PtrMYB3-DR and PtrMYB21-DR) were generated by fusing the full-length PtrMYB3 or PtrMYB21 cDNA in frame with the dominant EAR repression sequence Arabidopsis thaliana (ecotype Columbia) by agrobacterium-mediated transformation. For each construct, at least 60 transgenic Arabidopsis plants were generated for phenotypic analyses. The overexpression and dominant repression constructs were also introduced into poplar trees by agrobacterium-mediated transformation as described The PtrMYB overexpression constructs were created by ligating the full-length For light microscopy, stem segments embedded in low viscosity (Spurr's) resin (Electron Microscopy Sciences) were cut into 1-\u00b5m-thick sections with a microtome and stained with toluidine blue EF1\u03b1 reference gene in each sample. The efficiencies of the PCR reactions among the samples were compared and found to be very similar between the genes analyzed and the EF1\u03b1 reference gene, which range from 78% to 80%. The data were the average of three biological replicates.Total RNA was isolated from leaves with a Qiagen RNA isolation kit (Qiagen). First strand cDNAs were synthesized from the total RNA treated with DNase I and then used as a template for PCR analysis. The real-time quantitative PCR was performed with the QuantiTect SYBR Green PCR kit (Clontech) using first strand cDNAs as templates. The relative expression level of each gene was calculated by normalizing the PCR threshold cycle number of each gene with that of the To test the ability of tree MYBs to activate poplar secondary wall biosynthetic gene promoters Escherichia coli. The recombinant MYB-MBP protein was purified using amylose resin and then used for electrophoretic mobility shift assay (EMSA). The biotin-labeled SMRE oligonucleotides were incubated with 100 ng of MYB-MBP in the binding buffer . The MYB-bound DNA probes were separated from the unbound ones by polyacrylamide gel electrophoresis, electroblotted onto nitrocellulose membrane and detected by the chemiluminescent method The tree MYBs were fused in frame with the maltose-binding protein (MBP) and expressed in t test program , and the quantitative difference between the two groups of data for comparison in each experiment was found to be statistically significant (p<0.001).The experimental data of the quantitative PCR analysis and GUS activity assay were subjected to statistical analysis using the Student's The GenBank accession numbers for the genes used in this study are PtrMYB3 (KF148675), PtrMYB20 (KF148676), PtrMYB2 (KF148677), PtrMYB21 (KF148678), MYB46 (At5g12870), MYB83 (At3g08500), EgMYB2 (AJ576023), PtMYB4 (AY356371), MYB58 (At1g16490), MYB63 (At1g79180), OsMYB46 (JN634084), ZmMYB46 (JN634085), SbMYB46 (XP_002443268), HvMYB46 (AAU43823), BdMYB46 (XP_003575963), VvMYB46A (XP_002282821), VvMYB46B (XP_002275467), MtMYB46 (XP_003597423), GmMYB46A (XP_003542045), GmMYB46B (XP_003539482), GmMYB46C (XP_003539066), GmMYB46D (XP_003543900), GmMYB46E (XP_003555035), PtrMYB28 (XM_002307154), PtrMYB192 (XM_002310643), CesA4 (At5g44030), CesA7 (At5g17420), CesA8 (At4g18780), FRA8 (At2g28110), IRX8 (At5g54690), IRX9 (At2g37090), 4CL1 (At1g51680), CCoAOMT1 (At4g34050), PtrCesA4 (XM_002301820), PtrCesA7 (XM_002308376), PtrCesA8 (XM_002316779), PtrCesA17 (XM_002325086), PtrCesA18 (XM_002305024), PtrGT43A (JF518934), PtrGT43B (JF518935), PtrGT43C (JF518936), PtrGT43D (JF518937), PtrGT43E (JF518938), PtrGT47C (XM_002314266), PtrGT8D (XM_002319766), PtrGT8E (XM_002310744), PtrGT8F (XM_002326863), PtrCCoAOMT1 (XM_002313089), and PtrCOMT2 (XM_002317802)."} +{"text": "Global myocardial edema, a surrogate measure of myocardial inflammation, has been reported in patients with chronic dilated cardiomyopathy (DCM) based on T2-weighted STIR imaging. However, STIR imaging is prone to motion artefact and issues related to cardiac cycle timing and coil sensitivity. We hypothesised that a novel quantitative CMR T2 mapping technique would provide a reliable and reproducible measure for the detection of myocardial edema in chronic non-ischemic DCM.Consecutive DCM patients with disease duration greater than 3 months and age/sex matched healthy volunteers were prospectively enrolled. Exclusion criteria included coronary artery disease, systemic inflammatory disease, acute myocarditis or raised cardiac biomarkers. A mid-ventricular short axis slice was acquired using a T2-prepared single-shot SSFP readout [Ninety DCM , mean LVEF 41%) and 28 healthy volunteers , mean LVEF 68%) were studied. Subjects with DCM had a significantly higher myocardial T2 compared to healthy controls . T2 had good interobserver variability and interstudy reproducibility .Quantitative T2 mapping is highly reproducible. Myocardial T2 times are significantly higher in DCM patients than those in healthy controls, suggesting that myocardial inflammation is present in chronic DCM. Further studies are needed to assess the clinical and prognostic impact of these findings in the DCM population.This study was supported by the UK NIHR Cardiovascular Biomedical Research Unit of Royal Brompton Hospital and Imperial College, London. Dr Feng is a visiting academic supported by the National Basic Research Program of China (973 Program) (Grant No. 2010CB732502) and National Natural Science Funds of China (Grant No.30800254). Dr He was supported by Wellcome Trust Value In People (VIP) award and now holds a British Heart Foundation (BHF) Intermediate Basic Science Fellowship (FS/08/26225)."} +{"text": "Salmonella enterica serotype Typhimurium (S. Typhimurium) during colitis. However, the chemotaxis receptors conferring this fitness advantage and their cognate signals generated during inflammation remain unknown. Here we identify respiratory electron acceptors that are generated in the intestinal lumen as by-products of the host inflammatory response as in vivo signals for methyl-accepting chemotaxis proteins (MCPs). Three MCPs, including Trg, Tsr and Aer, enhanced the fitness of S. Typhimurium in a mouse colitis model. Aer mediated chemotaxis towards electron acceptors (energy taxis) in vitro and required tetrathionate respiration to confer a fitness advantage in vivo. Tsr mediated energy taxis towards nitrate but not towards tetrathionate in vitro and required nitrate respiration to confer a fitness advantage in vivo. These data suggest that the energy taxis receptors Tsr and Aer respond to distinct in vivo signals to confer a fitness advantage upon S. Typhimurium during inflammation by enabling this facultative anaerobic pathogen to seek out favorable spatial niches containing host-derived electron acceptors that boost its luminal growth.Chemotaxis enhances the fitness of Salmonella serotypes to ensure their transmission to a new host. Salmonella serotypes can use alternative electron acceptors that become available during colitis to support their growth by anaerobic respiration, therefore giving the pathogen an advantage over the fermenting microbiota. While it is known that S. Typhimurium benefits from chemotaxis and motility during colitis, the chemotaxis signals the pathogen detects in vivo have not been elucidated. Here we demonstrate that during intestinal inflammation, S. Typhimurium is able to seek energetically favorable niches by detecting the presence of nitrate and tetrathionate, two respiratory electron acceptors that are generated as by-products of the host inflammatory response. Through this mechanism, energy taxis provides a fitness advantage for S. Typhimurium, and likely other facultative anaerobic bacteria, during in vivo growth in the intestinal lumen.The ability to thrive in the inflamed gut is crucial for S. Typhimurium uses its virulence factors, two type III secretion systems (T3SS-1 and T3SS-2), to trigger acute intestinal inflammation S. Typhimurium in the lumen of the inflamed gut enhances its rate of transmission S. Typhimurium during colitis remain incompletely understood.The enteric pathogen S. Typhimurium in the lumen of the inflamed intestine S. Typhimurium to seek out favorable metabolic niches. It has been proposed that motility and chemotaxis enable S. Typhimurium to migrate towards complex polysaccharides available for fermentation. Galactose residues are present in high abundance at the cecal mucosa S. Typhimurium through Trg, a methyl-accepting chemotaxis protein (MCP) that senses galactose and ribose S. Typhimurium to get close to the mucosal surface and it has been hypothesized that a concentration gradient of galactose residues might be a possible chemotactic cue that attracts S. Typhimurium towards the intestinal epithelium S. Typhimurium to enhance its growth during intestinal inflammation. Mucus production in the gut is raised as part of the mucosal defense against S. Typhimurium infection Flagella-mediated motility is required for efficient growth of S. Typhimurium during colitis was warranted. To this end, we determined how mutations in genes encoding different MCPs affect growth in the inflamed intestine and performed follow-up studies to elucidate the mechanisms by which chemotaxis confers a growth advantage during intestinal inflammation.We thus reasoned that further investigation into the mechanisms by which chemotaxis enhances the fitness of S. Typhimurium. Preconditioning with streptomycin disrupts the resident microbiota and allows S. Typhimurium to elicit acute cecal inflammation, which is an animal model for human gastroenteritis (reviewed in S. Typhimurium during inflammation would be to determine which MCP(s) confer(s) a luminal growth advantage in the mouse colitis model. To this end we generated S. Typhimurium strains carrying mutations in mcpA (FR37), mcpB (FR36), mcpC (FR35), trg (FR42), aer (FR5) or tsr (FR4). As a positive control, we included a chemotaxis-deficient S. Typhimurium cheY mutant (FR13), which lacks a core chemotaxis component that is essential for signal transduction from MCPs to the flagellar apparatus. Each mutant was tested for its ability to compete with wild-type S. Typhimurium (IR715) for luminal growth in the mouse colitis model (competitive infection assay).We used the mouse colitis model S. Typhimurium wild type (IR715) and one of the above mutants resulted in acute cecal inflammation (Kc and Nos2 genes (Kc and Nos2 encode the inflammatory markers keratinocyte-derived cytokine (KC) and inducible nitric oxide synthase (iNOS), respectively. Wild-type S. Typhimurium (IR715) was recovered in significantly higher numbers (P<0.05) from the colon contents than a chemotaxis-deficient cheY mutant (FR13) four days after infection (S. Typhimurium in the inflamed gut trg mutant (FR42) was recovered in lower numbers than the wild type. In contrast, mutations in mcpA, mcpB or mcpC did not reduce the fitness of S. Typhimurium (S. Typhimurium wild type (IR715) was recovered in significantly (P<0.05) higher numbers from colon contents than an aer mutant (FR5) or a tsr mutant (FR4) (S. Typhimurium (IR715) over an aer tsr mutant (FR6) was not significantly greater than that over a tsr mutant (FR4).Infection of streptomycin pre-treated mice with the ammation and marks2 genes . Kc and nfection , which chimurium . Remarkant (FR4) , which iS. Typhimurium, aer is located upstream of mcpC, a gene encoding a MCP that is absent from Escherichia coliaer could be polar on expression of the downstream mcpC gene. However, inactivation of aer did not alter mcpC mRNA levels in vitro (mcpC could account for the phenotype of the aer mutant (FR5) in the mouse colitis model (mcpC did not reduce fitness (aer gene under the control of its native promoter on a low-copy number plasmid (pFR5) and introduced this plasmid into the aer mutant. Streptomycin pre-treated mice were infected with an equal mixture of the aer mutant carrying a control vector (pWSK129) and an aer mutant complemented with the cloned aer gene cloned on pFR5. The complemented aer mutant (aer[pFR5]) was recovered in significantly (P<0.05) higher numbers from colon contents than the aer mutant carrying a control vector (aer[pWSK129]) , therebyS. Typhimurium causes intestinal inflammation by using T3SS-1 for epithelial invasion and T3SS-2 for macrophage survival invA) and T3SS-2 (through a mutation in spiB) renders S. Typhimurium unable to trigger gut inflammation in the mouse colitis model invA spiB mutant (SPN452) and an invA spiB trg mutant (FR43). Mice infected with this mixture did not exhibit elevated expression of inflammatory markers and an invA spiB aer mutant (FR11) or an invA spiB mutant (SPN452) and an invA spiB tsr mutant (FR10). Mice infected with these mixtures neither developed intestinal pathology , an invA spiB mutant (SPN452) and an invA spiB aer mutant (FR11). Mice infected with this mixture developed acute intestinal inflammation, as indicated by elevated expression of inflammatory markers (invA spiB mutant was recovered in higher numbers than the invA spiB aer mutant (S. Typhimurium wild type generated an environment in which the aer gene conferred a fitness advantage upon the invA spiB mutant. Similar results were obtained when mice were infected with the S. Typhimurium wild type (ATCC14028), an invA spiB mutant (SPN452) and an invA spiB tsr mutant (FR10) . CollectS. Typhimurium during colitis has been linked to the transmigration of neutrophils into the intestinal lumen 2O32\u2212) generated by the colonic epithelium 4O62\u2212) S. Typhimurium from a point of inoculation into the surrounding motility agar was measured after anaerobic incubation for 6.5 hours. Halo formation was not observed with non-motile or chemotaxis-deficient strains. The tsr mutant (FR4) formed halos with significantly (P<0.05) reduced diameter compared to those produced by the S. Typhimurium wild type (IR715) both in the presence or in the absence of tetrathionate (P<0.05) larger diameter than an aer mutant (FR5) when tetrathionate was present in the motility agar, but not when tetrathionate was absent (aer mutant (FR5) or the wild type and a tsr mutant (FR4) indicated that inactivation of these chemotaxis receptors did not produce a growth defect ttrA) mutant to migrate towards tetrathionate in the motility plate assay. Consistent with our hypothesis, a ttrA mutant (SW661) formed halos with a significantly (P<0.05) smaller diameter than the wild type (IR715) when tetrathionate was present in the motility agar, but not when this terminal electron acceptor was absent and an aer mutant (FR5) were recovered in equal numbers from the capillary. In contrast, the wild type was recovered in significantly higher numbers when the capillary contained tetrathionate (aer gene (pFR5) into the aer mutant resulted in increased recovery from a capillary containing tetrathionate. These data provided further support for the idea that Aer mediates chemotaxis towards tetrathionate. When a capillary containing tetrathionate was submerged in a reservoir filled with a ttrA mutant and a ttrA aer mutant, both strains were recovered in equal numbers (in vitro. Consistent with the fact that genes for the utilization of inferior electron acceptors are repressed in the presence of oxygen aer open reading frame (C65T) leading to an amino acid substitution (S22F) that inactivates the FAD-binding domain of Aer S. Typhimurium wild type was recovered in significantly higher numbers than the S22Faer mutant from a tetrathionate-containing capillary (tsr mutant (FR4) were recovered in equal numbers from a capillary containing tetrathionate and a ttrA aer mutant (FR9) or a ttrA mutant (SW661) and a ttrA tsr mutant (FR8). The ttrA mutant (SW661) outcompeted the ttrA tsr mutant (FR8) (ttrA mutant (SW661) with the ttrA aer mutant (FR9). These data suggested that the ability of S. Typhimurium to utilize tetrathionate as an electron acceptor was required to see a benefit of Aer during growth in the inflamed gut. Finally, the S. Typhimurium wild type outcompeted the S22Faer mutant (FR47) in streptomycin pre-treated mice, suggesting that redox sensing by Aer conferred a fitness advantage in the mouse colitis model.To investigate whether the fitness advantage conferred by the nt (FR8) , suggestS. Typhimurium wild type (IR715) and an aer mutant (FR5) or with an equal mixture of a ttrA mutant (SW661) and a ttrA aer mutant (FR9). Wild-type S. Typhimurium was recovered in significantly higher numbers (P<0.05) from colon contents than an aer mutant at 10 days and 28 days after infection (P<0.05) reduced during competition of the ttrA mutant with the ttrA aer mutant. By day 28 after infection, strains defective for tetrathionate respiration (i.e. the ttrA mutant and the ttrA aer mutant) were no longer recovered from colon contents.To rule out that our phenotype was a result of disrupting the microbiota by treating mice with streptomycin, we infected genetically resistant (CBA) mice in the absence of streptomycin (mouse typhoid model). In this model, mice develop cecal inflammation by day 10 after infection nfection . At 10 d3\u2212), an electron acceptor that becomes available during inflammation tsr mutant (FR4) were recovered in equal numbers from the capillary. However, when the capillary contained 0.1 mM nitrate, the wild type was recovered in significantly higher numbers than the tsr mutant (tsr gene (pFR6) into the tsr mutant resulted in increased recovery from a capillary containing 0.1 mM nitrate. Interestingly, the wild type was recovered in similar numbers as an aer mutant from a capillary containing 0.1 mM nitrate. However, Aer was able to mediate energy taxis towards higher concentrations of nitrate, because the wild type was recovered in significantly higher numbers than an aer mutant from a capillary containing 1 mM nitrate. Neither Aer nor Tsr mediated energy taxis towards nitrate when the assay was repeated under aerobic conditions. In conclusion, Tsr, but not Aer, functioned in mediating energy taxis towards low concentrations of nitrate in an anaerobic environment.Inflammation induces expression of inducible nitric oxide synthase (iNOS), an enzyme catalyzing the production of nitric oxide (NO). Nitric oxide can react with ROS to generate nitrate and a napA narZ narG tsr mutant (FR46) were recovered in similar numbers from a capillary containing 0.1 mM nitrate and a napA narZ narG tsr mutant (FR46). The fitness advantage conferred by Tsr was abrogated during competition of the napA narZ narG mutant with the napA narZ narG tsr mutant and a S. Typhimurium wild-type strain that was marked by a phoN::Kan insertion to facilitate recovery (AJB715) (competitive infection assay). Remarkably, higher numbers (P<0.05) of the wild-type strain (AJB715) were found in close association with tissue, while equal numbers of both strains were recovered from the luminal fluid 8 hours after infection (invA spiB mutant (SPN452) and an invA spiB aer mutant (FR11) (P<0.05) reduced in loops infected with a mixture of the invA spiB mutant and the invA spiB aer mutant compared to loops infected with a mixture of the wild type and aer mutant (tsr mutant (FR4) from tissue-associated samples, but not from the luminal fluid of loops infected with a mixture of both strains (Tetrathionate respiration has recently been shown to enhance growth of nfection . No benet (FR11) . Measurer mutant . The wil strains . We concS. Typhimurium wild type (IR715), an aer mutant (FR5), a tsr mutant (FR4) or a chemotaxis-deficient mutant and organs were collected for analysis four days later. Mice infected with the S. Typhimurium wild type developed acute cecal inflammation reduced numbers from the colon contents of mice than wild-type S. Typhimurium , the ammation . There whimurium . Collecthimurium .S. Typhimurium resides in the lumen of the inflamed gut, but not during luminal growth in the non-inflamed intestine Motility and chemotaxis confer a fitness advantage when S. Typhimurium readily consumes these electron acceptors to boost its luminal growth in the inflamed gut S. Typhimurium to actively seek out niches containing these electron acceptors. Here we show that energy taxis enhances the fitness of S. Typhimurium in the inflamed intestine, by enabling the pathogen to migrate towards favorable metabolic niches , thereby altering the redox state of the cell. The flavin cofactor bound by Aer is proposed to sense changes in the redox state by becoming oxidized or reduced through interaction with a component of the electron transport chain, a process that generates the on and off signal for Aer-mediated energy taxis S. Typhimurium strains that were unable to respire tetrathionate neither exhibited Aer-mediated energy taxis towards tetrathionate in vitro and grown anaerobically at 37\u00b0C overnight. The cultures were spread on agar plates containing the appropriate antibiotics to determine the ratio of recovered bacteria. Three technical replicates were performed for each experiment and each experiment was repeated at least three times independently.To determine competitive indices (CI) after The plasmids and primers used in this study are listed in aer gene was amplified by PCR using the primers listed in aer PCR product was then cloned into the suicide plasmids pGP704 and pEP185.2 using the SalI and SacI restriction enzyme sites to construct pFR2 and pFR4, respectively. A fragment of the tsr gene was amplified by PCR and subcloned into pEP185.2 using the SalI and SacI restriction enzyme sites to create pFR3. The KSAC kanamycin resistance cassette of pBS34 was subcloned into the XbaI restriction site of pSPN57, generating a Kan-marked allelic exchange invA deletion construct (pSPN60).An internal fragment of the aer gene was constructed by using primers del_A and del_B to PCR amplify a 404 bp 5\u2032 flanking sequence that included 9 nucleotides of the aer coding region. Primers del_C and del_D were used to PCR amplify a 573 bp fragment that included 24 nucleotides of the 3\u2032 coding region. The two PCR fragments were joined using the splicing overlap extension (SOE) technique aer deletion was digested with BamHI and SalI and cloned into pRDH10 to generate plasmid pFR7.A suicide plasmid for generating a deletion of the aer open reading frame was constructed by PCR amplifying a 5\u2032 fragment of the aer gene using primer del_A, which anneals 395 bps upstream of the gene and primer S22F_B, which contains a missense mutation (C65T). Primer S22F_C, which encodes the same missense mutation and primer del_D were used to amplify the remaining aer ORF plus 549 bps downstream of the aer stop codon. The two PCR fragments were joined using the SOE technique aer allele with a C65T substitution was digested with BamHI and SalI and cloned into pRDH10 to generate pFR8.A suicide plasmid for introducing a point mutation into the aer mutant, the aer open reading frame including the promoter region was amplified using the primers listed in For complementation of the tsr mutant, the tsr open reading frame including the promoter region was amplified using the primers listed in For complementation of the E. coli DH5\u03b1 \u03bbpir and introduced into the S. Typhimurium strain IR715 by conjugation using E. coli S17-1 \u03bbpir as a donor strain. Transconjugants were selected on LB plates containing nalidixic acid (IR715) and antibiotics selecting for integration of the suicide plasmid. Integration of the suicide plasmids was verified by PCR. Using this methodology, pFR2, pFR4 and pFR3 were integrated into the aer and tsr genes of IR715 to yield strains FR5, FR7 and FR4, respectively. An mcpA mutant (MB193) mcpB mutant (QW111), mcpC mutant (SM19) and trg mutant (SM15) were constructed by the one-step mutagenesis method mcpC gene or the trg gene with a Cm cassette, generating SM19 and SM15, respectively. Primers were designed with overhang sequences complementarily to the beginning and end of Kan cassette in pKD4 to replace most of the mcpA or mcpB open reading frames with a Kan cassette to generate strains MB193 and QW111, respectively.Suicide plasmids were propagated in invA locus of IR715 was deleted with the KSAC Kan resistance cassette by conjugating pSPN60 from S17-1 \u03bbpir into IR715, selecting for double-crossover transconjugants on LB plates containing Nal and Kan. A strain positive for amplification with primers directed outwards from KSAC and primers directed inwards from beyond the flanking regions employed in the deletion of invA was labeled SPN453. An unmarked invA deletion mutant was then generated by conjugating pSPN57 from S17-1 \u03bbpir into SPN453 by following a previously described methodology invA deletion and negative for invA amplification was termed SPN455.The aer was deleted by conjugation of pFR7 from E. coli S17-1 \u03bbpir into S. Typhimurium strain IR715. Transconjugants were selected for on LB plates containing Nal and Cm. Sucrose counter-selection was performed as described previously s was verified by PCR to carry a deletion in aer and was denoted FR38.The aerS22F mutation was constructed by conjugation of pFR8 from E. coli S17-1 \u03bbpir into S. Typhimurium strain FR38. Transconjugants were selected for on LB plates containing Nal and Cm. Sucrose counter-selection was performed as described previously s, denoted FR47, was verified by PCR amplification and sequence analysis.The int-201 was used for generalized transduction. Transductants were routinely purified from phage contamination on Evans blue-Uranine agar and then cross-struck against P22 H5 to confirm phage sensitivity. The ttrA::pSW171 mutation from SW661 was introduced into FR4 and FR7 to yield strains FR8 and FR9, respectively. The tsr::pFR3 or aer::pFR2 mutations were introduced into the SPN452 to create strains FR10 and FR11, respectively. The cheY::Tn10 mutation from AT350 was transduced into IR715 to generate strain FR13. The mcpC::Cm mutation from SM19 was transduced into IR715 to generate FR35. The mcpB::Kan mutation from QW111 was transduced into IR715 to generate FR36. The mcpA::Kan mutation from MB193 was transduced into IR715 to generate FR37.The trg::Cm mutation from SM15 was transduced into IR715 to generate FR42.Phage P22 HT105/1 spiB deletion was generated in SPN455 by transducing the merodiploid \u0394spiB::pSPN56(\u0394spiB) locus from SPN458 followed by sucrose selection as previously described spiB deletion and negative for spiB amplification was dubbed SPN487. The trg::Cm mutation of SM15 was then transduced into SPN487, creating FR43.An unmarked Mouse and calf experiments adhered to USDA guidelines and were approved by the Institutional Animal Care and Use Committees at the University of California at Davis and Texas A&M University, respectively.S. Typhimurium in 0.1 ml LB broth as described previously 8 CFU of each strain. For single strain infections, animals were inoculated with approximately 1\u00d7109 CFU per animal. Animals were euthanized 4 days after infection and organs were collected for RNA purification and bacteriological analysis as described previously Female C57BL/6J mice, aged 9\u201312 weeks, were obtained from The Jackson Laboratory (Bar Harbor). Animals were pretreated with 20 mg Streptomycin and orally inoculated 24 hours later with 0.1 ml LB broth or 8 CFU of each bacterial strain. Animals were euthanized at the indicated time points to collect organs.Female CBA/J mice, aged 6\u20138 weeks, were obtained from The Jackson Laboratory (Bar Harbor). Animals were inoculated with 0.1 ml of a suspension (LB broth) containing an equal mixture of 5\u00d7108 CFU of each strain as indicated. 8 h after infection, loops were resected and luminal contents collected. Loops were opened longitudinally and 6 mm tissue biopsy punches were collected, and the homogenized tissue spread on selective media. Each bacterial strain was tested in at least three different animals. To facilitate recovery of Salmonella strains from biological samples, the wild-type strain (IR715) was marked with a mutation in the phoN gene (AJB715).Bovine ligated ileal loops surgery was performed as described previously Soft agar motility plates containing LB and 0.3% (w/v) agar or soft agar plates supplemented with 5 mM tetrathionate were inoculated with single colonies and incubated at 37\u00b0C anaerobically for 6.5 hours and the area of the halo was determined . Each experiment was performed in triplicate.600 between 0.6\u20130.8). An equal volume of each strain was added to a tube and washed twice in 0.5 ml of chemotaxis buffer , 0.1 mM EDTA). The pellet was resuspended in CB to a concentration of between 1\u00d7107 and 1\u00d7108 CFU/ml and 100 \u00b5l of the culture was added to a 1.5 ml tube. To make the capillaries, 1 \u00b5l glass capillaries that had been sealed on one end were briefly flamed and then added to 1.5 ml microcentrifuge tubes containing tetrathionate or nitrate at the indicated concentrations. The glass capillary was then placed into a 1.5 ml microcentrifuge tube containing S. Typhimurium strains and incubated at 37\u00b0C inside the anaerobe chamber for 45 minutes. The glass capillary was removed, and wiped off and dilutions of the contents of the capillary tube were spread on agar plates containing the appropriate antibiotics. Three technical replicates were performed for each experiment and each experiment was repeated at least three times independently.Capillary assays were performed as described previously Nos2 and Kc were normalized to mRNA levels of the housekeeping gene Gapdh.Eukaryotic gene expression was determined by real-time PCR as previously described mcpC in each sample was normalized to the respective levels of guanyl nucleotide kinase mRNA, encoded by the gmk gene.Bacterial RNA was isolated using the Aurum Total RNA Kit (Bio-Rad) according to the recommendations of the manufacturer. Afterward, an additional DNase treatment was performed using the Ambion DNA removal and inactivation kit. RT-PCR and real-time PCR were performed as described previously Formalin fixed cecal tissue sections were stained with hematoxylin and eosin, and a veterinary pathologist performed a blinded evaluation using criteria published previously t-test was used to determine whether differences in fold changes between groups were statistically significant (P<0.05). Significance of differences in histopathology scores was determined by a one-tailed non-parametric test (Mann-Whitney).Fold changes of ratios were transformed logarithmically prior to statistical analysis. An unpaired Student's Figure S1Inflammatory changes in the cecum in the mouse colitis model. For selected experiments shown in (PDF)Click here for additional data file.Figure S2Polarity of aer::pFR4 on expression of mcpC. (A) Schematic representation of the genetic region surrounding the aer gene. (B) Expression levels of mcpC were determined by quantitative real-time PCR with primers listed in aer (white bar) relative to mRNA levels in the S. Typhimurium wild-type (wt) strain (AJB715), which were set to 100%.(PDF)Click here for additional data file.Figure S3Anaerobic growth of S. Typhimurium strains on motility plates and in LB broth. (A) Halo size (arbitrary units) around a point of inoculation determined after 6.5 hours anaerobic incubation in the motility plate assay. Bars represent averages from three independent experiments \u00b1 standard error. *, P<0.05; ns, not significantly different; nh, no halo. (B) Representative images of halos produced in the motility plate assay. (C) Competitive growth of the indicated mixtures of S. Typhimurium strains in LB broth under anaerobic conditions in the absence (\u2212) or presence (+) of tetrathionate or nitrate . Bars represent geometric means of competitive indices (CI) from at least three independent experiments \u00b1 standard error.(PDF)Click here for additional data file.Figure S4Aer boosts growth in the mouse typhoid model. Groups (N\u200a=\u200a6) of genetically resistant CBA mice were inoculated with the S. Typhimurium wild-type strain (wt) and an aer mutant (aer) or with a ttrA mutant (ttrA) and a ttrA aer mutant (ttrA aer) and organs were collected for analysis on the indicated days after infection. Bars represent geometric means of competitive indices (CI) recovered from colon contents \u00b1 standard error. *, P<0.05; NR, no bacteria recovered.(PDF)Click here for additional data file.Figure S5Analysis of histopathological lesions induced by infection with different S. Typhimurium strains. Groups of streptomycin pre-treated mice (N\u200a=\u200ais indicated in panel A) were inoculated with sterile medium (mock-infected) or with the S. Typhimurium wild-type strain (wt), an aer mutant (aer) or a cheY mutant (cheY) and organs were collected for analysis four days after infection. (A) Blinded histopathology scoring of cecal inflammation showing averages (bars) of scores for individual animals (circles). (B) Representative images of histopathological changes. (C) Expression of pro-inflammatory markers in the cecal mucosa was determined by quantitative real-time PCR analysis. Bars represent geometric means of Kc and Nos2 mRNA copy numbers as fold-change over mRNA levels in mock-infected mice \u00b1 standard error.(PDF)Click here for additional data file.Figure S6Model for the mechanism by which energy taxis confers a fitness advantage during colitis. Upon ingestion, a fraction of the luminal S. Typhimurium population migrates along a concentration gradient of galactose residues towards the mucus layer S. Typhimurium population now uses energy taxis to migrate towards environments containing tetrathionate or nitrate and subsequently uses anaerobic respiration to gain a fitness advantage over competing microbes that grow by fermentation.(PDF)Click here for additional data file.Table S1Calculation of competitive indices for experiments using the capillary assay. S, strain number; I, inoculum; G.M, geometric mean; C/ml, colony forming units per ml; CFU/cap, geometric mean of colony forming units per capillary after incubation; IR, input ratio; OR, output ratio; CI, competitive indices; NR, napA narZ narG mutant; T (5 mM), capillary containing 5 mM tetrathionate; B, capillary containing buffer; N (1 mM), capillary containing 1 mM nitrate; (O2), experiment was performed under aerobic conditions.(PDF)Click here for additional data file."} +{"text": "To the Editor: Peste des petits ruminants virus causes severe infectious disease in sheep and goats in Africa and Asia. Pneumo-enteritis clinical signs are dominated by ocular and nasal discharge, and mortality rates are high (wilayas), which covered 99.3% of the national sheep and goat stocks (mougataas), 21 were randomly selected. A single geographic point was randomly sampled within each of the selected mougataas, and 100 small ruminants were sampled in a 7-km radius around the coordinates. None of the mougataas in which sampling occurred had a PPRV vaccination program.A seroprevalence survey was implemented in October 2010 to assess PPRV spatial distribution in Mauritania. The study was limited to 8 southern provinces during January\u2013March 2012. All outbreaks were investigated, and biologic samples were collected for laboratory diagnostics. <45% were considered positive. A logistic beta-binomial regression model was used to analyze prevalence rates within mougataas. Swab samples were tested by using reverse transcription PCR (RT-PCR) adapted to a 1-step format and based on nucleoprotein (NP) 3\u2013NP4 PPRV-specific primers targeting the 3\u2032 end of the NP gene . Optical density values were converted to inhibition percentages; according to the ELISA cutoff value, inhibition percentages of NP gene .\u22126) . PPRV infection was widespread: prevalence rates ranged from 3% (Guerou) to 98% (Kobeni) Figure 2\u22126) Table. T\u22126) Figure 2Three suspected outbreaks of PPR were reported during January\u2013March 2012 Figure 2N-gene sequences were obtained from 2 sheep swab specimens collected in Trarza during the outbreak survey in early 2012 (deposited in the GenBank under accession nos. KF483658 [Mauritania1_2012] and KF483659 [Mauritania5_2012]). These isolates were placed in a phylogenetic tree built from PPRV sequences recently collected in western and northern Africa (Morocco), as well as isolates from other parts of the world retrieved from GenBank. Phylogenetic analysis involved 255 nt located on the C terminus end of the NP gene of the virus (84 aa). The PPRV strain from Mauritania belonged to lineage II . SequencOur study results highlight 2 PPRV epidemiologic systems: northern Africa, where all identified PPRVs belonged to lineage IV and were closely related to PPRV initially identified in Sudan (Small ruminant density in Mauritania, peste des petits ruminants virus seroprevalence rates in 2010, and reported peste des petits ruminants outbreaks in early 2012."} +{"text": "Clinical and in vitro studies suggest that subchondral bone sclerosis due to abnormal osteoblast (Ob) functions, is involved in the progression and/or onset of osteoarthritis (OA). Human OA subchondral Ob show a differentiated phenotype, however they fail to mineralize normally. The canonical Wnt/b-catenin signaling pathway (cWnt) plays a key role in osteogenesis by promoting the differentiation and mineralization of Ob. Dickkopfs (DKKs) are potent antagonists whereas R-spondins (Rspo) are newly described agonists that play key roles in cWnt signalling. However, the regulation of DKKs and Rspos in OA Ob remains unknown.We prepared primary human subchondral Ob using the sclerotic medial portion of the tibial plateaus of OA patients undergoing knee arthroplasty, or from tibial plateaus of normal individuals at autopsy. DKK1, DKK2, SOST and Rspo-1 and -2 expression and production were evaluated by qRT-PCR and WB analysis. The regulation of their expression was determined in response to transforming growth factor-\u00df1 (TGF-\u00df1) and as a function of the growth of OA Ob. Selective inhibition was performed using siRNA techniques. cWnt signaling was evaluated by measuring target gene expression using the TOPflash Tcf/lef luciferase reporter assay and intracellular \u00df-catenin levels by WB. Mineralization was evaluated by Alizarin red staining. TGF-\u00df1 levels were determined by ELISA.DKK2 expression and production were elevated in OA Ob compared to normal whereas DKK1 was similar. Rspo2 expression was reduced in OA Ob whereas Rspo1 was similar. TGF-\u00df1mRNA expression and protein levels were high in OA Ob. TGF-b1 stimulated DKK2 expression and production in Ob whereas it inhibited Rspo2 expression. cWnt signaling was reduced in OA compared to normal Ob. This inhibition was due in part to elevated DKK2 levels and to reduced Rspo-2 levels since correcting DKK2 by siRNA or the addition of Rspo-2 increased cWnt signaling using the TOPflash reporter assay. These treatments also increased \u00df-catenin levels in OA Ob. Mineralization of OA Ob was reduced compared to normal Ob and was also corrected in part by inhibiting DKK2 or by Rspo2 addition. Both elevated DKK2 and reduced Rspo2 levels contributed to abnormal expression of bone markers by OA Ob.These studies demonstrate that elevated antagonist or reduced agonist levels of cWnt signalling interfere in normal Ob function and lead to abnormal mineralization. Since these are secreted soluble proteins, this could lead to potential new avenues of treatment of OA to correct their abnormal bone phenotype and mineralization."} +{"text": "Continuous glucose monitoring (CGM) in ICUs has the potential to improve patient safety and outcomes. The GluCath Intravascular CGM System uses a novel quenched chemical fluorescence sensing mechanism to measure glucose concentration (BG) in venous or arterial blood. This is the first report of its use in cardiac surgery patients.in vivo calibration 30 minutes later. Clinical staff managed blood glucose according to usual protocols. Glucose values were recorded each minute for 24 hours; hourly reference samples from the same arterial catheter were analyzed on a Radiometer ABL Blood Gas Analyzer.This ongoing clinical study is evaluating the system deployed via a standard 20G radial artery catheter inserted for routine care in 20 patients undergoing cardiac surgery. Data are presented from five run-in patients. Outcome measures are qualitative and quantitative . Sensors were inserted shortly after ICU admission with placement confirmed by ultrasound and The sensor was successfully deployed in all five patients and did not interfere with clinical care, blood pressure monitoring or sampling. One patient suffered a cardiopulmonary arrest; the sensor functioned successfully during resuscitation and urgent return to the operating room. One hundred and twenty reference samples ranging from 5.9 to 13.4 mmol/l were collected; 107/120 (89.2%) of GluCath measurements met ISO 15197 criteria (within \u00b120% of reference when BG >4.2 mmol/l; Figure in vivo calibration.The GluCath System measured glucose concentration continuously in a cardiac surgery ICU without compromising arterial line function or patient care. In all patients the sensor operated without interruption for 24 hours following a single"} +{"text": "Shwachman\u2013Diamond syndrome (SDS) is an autosomal recessive disorder (OMIM 260400), characterized by exocrine pancreatic insufficiency, skeletal abnormalities and bone marrow (BM) dysfunction, with a risk, as high as 30%, to develop myelodysplastic syndrome and/or acute myeloid leukaemia (MDS/AML). The SBDS gene (OMIM 607744) is localized on chromosome 7 at the band q11 and mutations of this gene are found in 90% of patients.SBDS gene was performed on an ABI Prism 3100 Genetic Analyzer using a BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems). The forward (F) and reverse (R) primers for amplifying exon 2 are AAATGGTAAGGCAAATACGG (2Fa), AGACCTCGATGAAGTTCT-GC (2Fb) and ACCAAGTTCTTTATTATTAGAAGTGAC (2R).Direct sequencing of whole exon 2 and flanking intronic regions of the We described a 14 years old boy who presented with skeletal system abnormalities , short stature, chronic dydpepsia, neutropenia and thrombocytopenia. Abdominal CT of patient showed congenital lipomatosis of pancreas and spine bone density shows osteoporosis. Direct sequencing for whole exons including intron-exon boundaries of patient showed two heterozygous mutations, c. [183_184TA>CT] + [258+2T>C].We treated the patient with pancreatin and Dicamax . We has observed for hematologic abnormality and prepared for bone marrow transplantation.We report a child with diverse clinical manifestations of SDS including short stature, chronic dydpepsia, skeletal system abnormalities, and neutropenia; the clinical diagnosis was confirmed by genetic analysis for the second time in Korea."} +{"text": "Rhesus monkey rhadinovirus (RRV) is a gamma-2 herpesvirus closely related to human herpesvirus 8 (HHV8). RRV encodes viral FLICE inhibitory protein (vFLIP), which has death effector domains. Little is known about RRV vFLIP. This study intended to examine its function in apoptosis. Here we found that RRV vFLIP inhibits apoptosis induced by tumor necrosis factor-\u03b1 (TNF-\u03b1) and cycloheximide. In HeLa cells with vFLIP expression, the cleavage of poly [ADP-ribose] polymerase 1 (PARP-1) and activities of caspase 3, 7, and 9 were much lower than those in controls. Cell viability of HeLa cells with vFLIP expression was significantly higher than control cells after apoptosis induction. However, RRV vFLIP appears unable to induce NF-\u03baB signaling when tested using NF-\u03baB reporter assay. RRV vFLIP was able to enhance cell survival under starved conditions or apoptosis induction. At early time points after apoptosis induction, autophagosome formation was enhanced and LC3-II level was elevated in cells with vFLIP and, when autophagy was blocked with chemical inhibitors, these cells underwent apoptosis. Full length of vFLIP is needed for the function against apoptosis as truncation variants of vFLIP were unable to block apoptosis induction. Moreover, RRV latent infection of BJAB B-lymphoblastoid cells protects the cells against apoptosis by enhancing autophagy to maintain cell survival. Knockdown of vFLIP expression in the RRV-infected BJAB cells with siRNA abolished the protection against apoptosis. These findings indicate that vFLIP protects cells against apoptosis by enhancing autophagosome formation to extend cell survival. The finding of vFLIP\u2019s inhibition of apoptosis via the autophagy pathway provides insights of vFLIP in RRV pathogenesis."} +{"text": "To assess cellular effects of celastrol and to identify target proteins as biomarkers for monitoring treatment regimes, we performed large-scale quantitative proteomics in cultured human lymphoblastoid cells, a cell type that can be readily prepared from human blood samples. Celastrol substantially modified the proteome composition and 158 of the close to 1800 proteins with robust quantitation showed at least a 1.5 fold change in protein levels. Up-regulated proteins play key roles in cytoprotection with a prominent group involved in quality control and processing of proteins traversing the endoplasmic reticulum. Increased levels of proteins essential for the cellular protection against oxidative stress including heme oxygenase 1, several peroxiredoxins and thioredoxins as well as proteins involved in the control of iron homeostasis were also observed. Specific analysis of the mitochondrial proteome strongly indicated that the mitochondrial association of certain antioxidant defense and apoptosis-regulating proteins increased in cells exposed to celastrol. Analysis of selected mRNA transcripts showed that celastrol activated several different stress response pathways and dose response studies furthermore showed that continuous exposure to sub-micromolar concentrations of celastrol is associated with reduced cellular viability and proliferation. The extensive catalog of regulated proteins presented here identifies numerous cellular effects of celastrol and constitutes a valuable biomarker tool for the development and monitoration of disease treatment strategies.Celastrol, a natural substance isolated from plant extracts used in traditional Chinese medicine, has been extensively investigated as a possible drug for treatment of cancer, autoimmune diseases, and protein misfolding disorders. Although studies focusing on celastrol's effects in specific cellular pathways have revealed a considerable number of targets in a diverse array of Tripterygium wilfordii) have been used in traditional Chinese medicine to treat fever, inflammation, and various other health problems in man in vitro and suppresses tumor growth in animal cancer models, as reviewed in Plants used in traditional Chinese medicine are rich sources of biologically active substances with potential therapeutic effects towards many human diseases HSPA1B (a well-known celastrol targets encoding the cytosolic heat shock protein Hsp70) and HSPD1 was used to differentially label the proteomes of two cell populations, which subsequently were incubated in either 0.8 \u00b5M celastrol (treated) or in vehicle alone for 24 h . This dootein 60 . Althougifically . RelativThe cellular proteome was defined from three replicated experiments in which 2765 different proteins were quantified in total . The coeIn the mitochondrial proteome analysis, 498 different quantified proteins were classified as mitochondrial by their listing in Human Mitocarta We assessed whether specific biological characteristics apply to the celastrol-regulated proteins by using the functional annotations of genes provided by the Gene ontology (GO) consortium To assign functional properties specifically to the celastrol-regulated proteins (cellular core proteins with ratio different from 1 by t-test), we performed a focused comparison of their GO annotation with the annotation of all quantified proteins. Proteins up-regulated by celastrol were overrepresented in 19 different \u201cbiological process\u201d GO categories , indicatTo analyze effects at the level of single proteins we manually grouped the quantified proteins into functional classes and 3 baHSPA1B) and Hsp27 (HSPB1) heat shock proteins, which are well-documented targets induced through activation of the heat shock response HSPD1 gene encoding Hsp60 contains regulatory regions for binding of transcription factors controlling both the heat shock response Celastrol up-regulated the cytoplasmic Hsp70 (HSPA5), Grp94 (HSP90B1), calnexin (CANX), calreticulin (CALR), ERp29 (ERP29), multiple protein disulfide-isomerases, as well as several glucosidase and glycosyltransferase enzymes involved in adding and trimming of sugar residues on ER-traversing glycoproteins. The cellular expression of several of these ER proteins is under control of the ER unfolded protein response, a well-described stress response activated by accumulation of misfolded proteins and various other cell stressors. The ER unfolded protein response signaling cascade initiates multiple events to restore ER protein homeostasis and function, including attenuation of protein translation to reduce the load of newly synthesized proteins on the ER and transcriptional activation of genes involved in restoring normal ER function, such as molecular chaperones A long list of up-regulated proteins involved in ER protein quality control and handling activities such as folding, maturation, and sorting of proteins destined for secretion was identified . These iProteins involved in the cellular defense against oxidative stress represented another well-defined group of celastrol-induced proteins that incHMOX1) was the single most (25 fold) highly up-regulated protein in our study and this protein has recently also been identified as a celastrol target in vascular smooth muscle cells The HO-1 , CLPP , HMOX1 (antioxidant response), and HSPA1B, HSPD1 (heat shock response). We used 1 \u00b5M celastrol to induce a strong transcriptional response has furthermore demonstrated that celastrol triggers the yeast antioxidant responses through activation of the yeast antioxidant transcription factor Yap1 We investigated whether changes in protein levels were due to transcriptional regulation by analyzing the mRNA levels of selected genes by quantitative reverse-transcriptase PCR (RT-PCR). We focused on genes involved in various cellular stress responses, including onse see and measA levels correspoA levels . This isCLPP mRNA . The target genes are thus transcribed at a high level several hours prior to the time of proteome analysis.We established that 8 h incubation was sufficient to induce the selected stress genes and furtGAPDH). We found lowered mRNA level from several genes involved in various cellular functions including ACTB (encoding the beta-actin protein) in treated cells. Even the mRNA levels from the highly induced HSPA1B and HSPD1 genes declined at longer exposure times , ACADM, re times . This inThe celastrol dose (0.8 \u00b5M) used throughout the proteomics studies was selected to give large expression level changes and to facilitate identification of a high number of celastrol-regulated proteins. Using the MTT metabolic activity assay, we found that 0.8 \u00b5M celastrol significantly reduced cellular viability. The viability decreased in a concentration-dependent manner reaching a maximum inhibitory effect at 1 \u00b5M with 24 h exposure time . We furtPRDX1), several ribosomal proteins, hexokinase 1 (HK1), BAX, and L-lactate dehydrogenase B (LDHB) than when analyzed in the cellular proteome experiments. From cross reference of the quantitative data for the 253 core proteins listed in MitoCarta that were quantified in both the mitochondrial and cellular proteome experiments, we identified more than 20 putative mitochondria translocating proteins . These iB (LDHB) . Some ofIt is clear from this and previous studies The mechanism by which celastrol activates multiple cellular pathways is still uncertain, but its ability to react with thiol groups of regulatory proteins possibly plays a key role Tripterygium wilfordii with potential anti-cancer activities, has recently been reported. Triptolide modifies the activity of the transcription factor TFII through specific and covalent binding to the XPB subunit in vivo cancer models before initiation of clinical trails. These investigations should include careful evaluation of pharmacokinetic parameters as we have observed cell specific differences in celastrol dose response with regard to both the toxicity and stress response induction and lysine (Lys6: [13C6] L-Lysine). Celastrol was added to a final concentration of 0.8 \u00b5M to cells grown in heavy isotopic media for 6 cell-doublings and equivalent amount of vehicle alone (dimethyl sulfoxide) was added to cells grown in standard isotopic media. Twenty-four hours later, celastrol-treated and untreated cells were collected and proteins were extracted according to two different protocols for analysis of either the cellular proteome or the mitochondrial sub-proteome were grown at 37\u00b0C in 95% humidified air and 5% (v/v) CO2 in culture flasks (TPP). Culture media for SILAC labeling were prepared according to guidelines in For analysis of the cellular proteome, equal amounts of protein coupled to mass spectrometry through a nano-electrospray source (Proxeon) essentially as described previously Mascot version 2.2.04 (Matrix Science) was used for peptide identification and MaxQuant version 1.0.13.13 The distribution of the up- and down-regulated proteins into GO categories was compared pairwise to the distribution of all quantified members of the cellular proteome study analyzed by Pearson Chi-Square statistics using the web-based WeGo tool RNA isolated from cells using the Total RNA isolation system (Promega) was analyzed by quantitative reverse-transcriptase PCR. RNA was transcribed into cDNA and analyzed by real-time PCR using Taqman probe chemistry (Applied Biosystems). Relative quantification with GAPDH as the reference gene was performed as previously described For Western blotting, soluble proteins extracted from cells by a detergent-based lysis buffer and centrifugation were transferred to a polyvinylidene fluoride membrane (Millipore) using a semidry blotting system (Bio-Rad). Proteins were detected by primary antibodies against the proteins Hsp60 , HO-1 , and Hsp70 using infrared dye-labeled secondary antibodies and an infrared imaging system .Cellular viability was addressed by the cellular reduction of MTT -2,5-diphenyltetrazolium bromide) assay Figure S1Celastrol dose response in lymphoblastoid cells evaluated by HSPA1B and HSPD1 mRNA levels. The dose-dependent induction of HSPA1B and HSPD1 mRNA were analyzed by quantitative RT-PCR using RNA isolated from lymphoblastoid cells treated with varying celastrol concentrations for 24 h. The relative mRNA levels from different genes were normalized using GAPDH mRNA as reference and presented relative to untreated control cells (t\u200a=\u200a0). Data are mean of two independent experiments (errors bars show the range) and each cDNA was analyzed in triplicate PCR reactions.(TIF)Click here for additional data file.Figure S2Variation between replicates in the cellular proteome study. Coefficient of variation for the quantitative ratios (treated/untreated) for the 1779 cellular core proteins illustrated by frequency histogram.(TIF)Click here for additional data file.Figure S3Cell specific celastrol toxicity and induction of heat shock gene expression. (A) Comparison of the dose dependent toxicity of celastrol in HEK293 and lymphoblastoid cells measured by the MTT assay. Viability of cells incubated for 24 h in celastrol expressed relative to cells incubated in vehicle alone (DMSO). (B) The inducible expression of markers of the heat shock response (HSPA1B) and ER UPR (DDIT3) in HEK293 following 24 h incubation in celastrol analyzed by quantitative RT-PCR as described in (TIF)Click here for additional data file.Table S1All quantified cellular proteins. This table lists the treated/untreated quantitative ratios for all 2765 different proteins quantified in at least one of three replicated cellular proteome experiments.(XLS)Click here for additional data file.Table S2Cellular core proteins. This table shows selected data (including the quantitative ratio (treated/untreated) for single experiments, mean ratio, and descriptive statistic) for the 1779 cellular core proteins quantified repeatedly in three cellular proteome experiments.(XLS)Click here for additional data file.Table S3Cellular core proteins 1.5 fold. This table lists the 158 cellular core proteins that are altered at least 1.5 fold.(XLS)Click here for additional data file.Table S4All mitochondrial proteins. This table lists the treated/untreated quantitative ratio for the 498 quantified proteins that are classified as mitochondrial by their listing in Human MitoCarta.(XLS)Click here for additional data file.Table S5Mitochondrial core proteins. This table shows selected data (including the quantitative ratio (treated/untreated) for single experiments, mean ratio, and descriptive statistic) for the 374 mitochondrial core proteins quantified repeatedly in three mitochondrial proteome experiments.(XLS)Click here for additional data file.Table S6Mito core proteins 1.5 fold. This table lists the 33 mitochondrial core proteins that are altered at least 1.5 fold.(XLS)Click here for additional data file.Table S7Protein evidence files. The table shows output from Maxquant's \u201cProtein Groups\u201d and contains data for all six experiments, three cellular proteome experiments , and three mitochondrial proteome experiments . Numbers in the \u201cPeptide ID\u201d column refers to (XLS)Click here for additional data file.Table S8Peptide evidence files. This table shows peptide evidence output from MaxQuant for identified proteins in (XLS)Click here for additional data file.Text S1Supplementary materials and methods. Detailed description of selected (DOC)Click here for additional data file."} +{"text": "Septic patients frequently develop critical illness myopathies (CIMs) that may represent a crucial factor for prolonged intensive care unit treatment and for ventilator weaning delay. Experimental findings have identified that oxidative stress plays a role in causing muscle depletion in chronic pathological states like sepsis. It is well documented that regular moderate physical exercise can decreased oxidative stress and enhance antioxidant functions.To investigate whether exercise training reduces oxidative damage in septic rats induced by cecal ligation and perforation (CLP).Wistar rats were randomly assigned to three groups: Sham (submitted to a fake surgery), CLP, and CLP that was previously trained (CLPT). The exercise training protocol consisted of 8 weeks of running on a treadmill, 5 days/week, for 60 minutes at 60% of the maximal running speed obtained on the graded treadmill test. Rats were subjected to CLP surgery; after 120 hours of surgical procedure they were killed by decapitation. Oxidative damage of lipids (thiobarbituric acid reactive species (TBARS)) and proteins (carbonyl groups) were analyzed in Soleus (type I fiber) and plantaris (type II fiber) muscles.See Table 1P = 0.06 (Table TBARS and carbonyl analysis for CLPT are lower than for CLP with statistical significance, except for carbonyl plantaris with 06 Table . Our dat"} +{"text": "LPIN2 gene located on the short arm of chromosome 18 have been identified as being responsible for the Majeed syndrome.Majeed syndrome is a rare, syndromic form of chronic recurrent multifocal osteomyelitis (CRMO) first described in 1989. The syndrome starts during infancy with recurrent relapses of osteomyelitis typically associated with fever, congenital dyserythropoietic anemia (CDA) and often neutrophilic dermatosis (Sweet syndrome). Homozygous mutations in the LPIN2 gene detected in two brothers with Majeed syndrome and to describe the clinical characteristics and response to treatment.To report a novel mutation in the LPIN2 gene re-sequencing of each affected child revealed a homozygous 2 bp deletion (c.1312_1313delCT) resulting in an early truncation of the protein (L438fs+16X), which confirmed the diagnosis in both patients. Clinically, both were refractory to the treatment with corticosteroids and TNF\u03b1 inhibition (etanercept). Elevated plasma levels of proinflammatory cytokines did not change significantly during the treatment. In M, rapid clinical and laboratory improvement was observed after introduction with anakinra (1.7 mg/kg/d), which has not yet been introduced to Y.Two Turkish brothers (13 (Y) and 29 (M) months old) with consanguinity of the parents were admitted with relapsing episodes of pain and \u2018pseudoparalysis\u2019 of upper and lower extremities since the age of 3 and 6 months, respectively. No concomitant fever has occurred during the attacks. Whole body MRI of the elder brother (M) revealed osteomyelitic changes of the metaphyses of tibiae, left fibula and left radius. Biopsy from lesions showed no malignancy and negative bacterial cultures. Both showed significant hypersedimentation (ESR 92 mm/hr (Y) and 96 mm/hr (M)), slight thrombocytosis, and moderate anemia (Hb 9.0 (Y) and 9.7 (M) g/dl). Bone marrow aspiration was consistent with congenital dyserythropoietic anemia (CDA) with 6% - 9% bi- or multinucleated erythrocytes. LPIN2 gene in two Turkish brothers with Majeed syndrome. Although our patients also presented with CDA none of them had fever during the attacks nor dermatological changes unlike previously described patients with Majeed. IL-1 inhibition showed promising clinical and laboratory results.We describe a novel mutation of the"} +{"text": "RNA interference can be mediated by fully complementary siRNA or partially complementary miRNA. siRNAs are widely used to suppress viral replication and the fully complementary siRNA bound Ago-2 in the RISC is known to degrade the target RNA. Although other argonaute proteins lacking slicer activity can also bind oligonucleotides with both si and miRNA structures, whether they can also contribute to antiviral effects is not entirely clear. We tested si and miRNA structured oligos for target repression in dual luciferase assays as well as for inhibition of Dengue and West Nile virus replication in ES cells expressing individual Ago proteins. In luciferase assays, both fully complementary and partially complementary oligos effectively repressed their targets in all individual Ago expressing cell lines, although the efficacy with fully complementary oligos was higher in Ago-2+ cells. However, partially complementary oligos had no effect on virus replication in any cell line, while fully complementary siRNAs were highly effective in Ago-2 expressing, but not in cells expressing other Ago proteins. This occurred irrespective of whether the target sequences were located in the coding region or 3\u2032UTR of the virus. We conclude that Ago-2 slicer activity is essential for anti-viral efficacy of siRNAs and miRNA-mediated translational repression/transcript destabilization is too weak to suppress the abundantly expressed flaviviral proteins. In either case, the 21\u201323 nucleotide dsRNAs associate in the cytoplasm with the RNA-induced silencing complex (RISC), whereupon one of the two RNA strands (passenger strand) is discarded and the other guide strand guides the RISC to mediate sequence-specific degradation of the corresponding mRNA (in the case of siRNAs) and/or translational repression/transcript destabilization by binding to the 3\u2032 untranslated region (UTR) (in the case of miRNAs) RNA interference (RNAi) is a phenomenon where small double stranded RNAs mediate sequence-specific regulation of gene expression. Essentially, RNAi can be induced either by endogenously encoded small RNAs called microRNAs (miRNAs), endogenously generated small interfering siRNAs (siRNA) or exogenously introduced siRNAs While siRNAs are designed to be fully complementary to the coding region of the mRNA target to be silenced, miRNAs are imperfectly complementary, generally having sequence matches in the 5\u2032 2-8nt seed sequence to the 3\u2032UTR of the target mRNA. Also, while siRNAs primarily degrade the target mRNA, miRNAs lead to translational repression/mRNA destabilization in the renilla luciferase 3\u2032UTR. This plasmid was transfected along with fully complementary or partially complementary dengue virus siRNA , whereas endogenous miRNAs generally target the 3\u2032UTR sequences. To test if this is indeed the case, we also tested si/miRNA resembling oligos that target the viral 3\u2032 UTR. Initially we tested several potential siRNAs targeting West Nile virus 3\u2032 UTR and identified one that potently suppressed viral replication in HeLa cells (not shown). We used this siRNA and its counterpart miRNA structure for viruWe also tested 2 additional siRNAs that we had earlier identified as potent suppressors of West Nile virus (WNV) Many of the proteins involved in the RNAi pathway accumulate in discrete cytoplasmic bodies called the processing or P bodies Our results suggest that although effective in reporter assays, completely complementary siRNAs loaded to Agos1, 3 and 4 and siRNAs mimicking miRNAs with only seed matches to the viral targets in the coding region or 3\u2032UTR loaded to any of the Agos fail to suppress an acute flaviviral infection.Because only Ago-2 has slicer activity and also because extensive pairing with the target is required for slicing, siRNAs bound to Ago-2 can degrade the target mRNA whereas siRNAs bound to Agos 1, 3 and 4 as well as miRNAs bound to any Ago protein can only mediate target repression by translational repression/transcript destabilization Many viruses also encode miRNAs that are completely or partially homologous with their viral targets and cellular miRNAs have also been proposed to affect viral replication Many of the proteins involved in the RNAi pathway accumulate in P bodies In summary, robust suppression of viral infection can only be achieved by completely complementary siRNA bound Ago-2-mediated target cleavage and miRNA mediated translational repression/mRNA destabilization or other Ago proteins appear to have no role in the anti-flaviviral RNAi.D (siRNA against dengue virus) ggatgtggattatttggaa; miFvED (miRNA mimic) ggatgtggtaaatttggaa; siWN-3UTR (siRNA against WNV 3\u2033 UTR) tgtagtgttcatagcaatt; miWN-3UTR (miRNA mimic against WN 3\u2032UTR) tgtagtgtagttagcaatt; siFvEJW (siRNA against WNV) gggagcattgacacatgtgca; si6 (siRNA against WNV), ctgtgacattggagagtca; siDV1 (negative control siRNA) cagcatattgacgctggga; siGFP (negative control siRNA) ggctacgtccaggagcgca.SiRNA and miRNA mimicking oligos were obtained from Dharmacon . The sense strand sequences from 5\u2032 to 3\u2032 end were: siFvED) target ggatgtggattatttggaa was synthesized at IDT.Reporter was constructed by cloning the DNA oligos containing the target sequence between the XhoI and NotI sites downstream of the Renilla luciferase gene in the psiCHECK2 vector . The DNA oligos for the dengue siRNA (siFvE5 cells/ml and seeded in 96 well plates in a volume of 100 \u00b5L/well. Reporter plasmids (20 ng of psiCHECK2 plasmid harboring the target region) together with siRNA or miRNA structured oligos (1 pmol siRNA/miRNA oligos) were cotransfected using Lipofectamine 2000 . After 24 h, Renilla and firefly luciferase activities were measured according to the manufacturer's instructions using the Dual-Glo luciferase assay kit. The ratio of Renilla to firefly luminescence was normalized to the negative control siRNA (siGFP or siDV1). Luminescence activity values represented an average of four replicates.Ago1-4 expressing ES cells and HeLa cells were cultured as described previously 4 cells per well one day before transfection. The siRNAs (8 pmol) were transfected into cells with RNAiMax (Invitrogen) per the manufacturer's instructions. 24 hours after transfection, the cells were infected with Dengue-2 or West Nile virus (moi\u200a=\u200a1-5). 72 hours later, the cells were stained with anti-flavivirus envelope specific antibody , followed by flow cytometric analysis to determine inhibition of virus replication.The antiviral activity of si/miRNAs was tested as previously described WNV replicon pWIIREPG-Z Untransfected, WNV replicon transfected and WNV or Dengue virus infected HeLa cells were immunostained for P bodies. Cells grown on cover slips were washed in PBS, fixed in 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, and blocked . The cells were then incubated with a rabbit anti -DCP1a antibody (Abcam) and West Nile or DENV-2 specific antibodies , followed by incubation with appropriate secondary antibodies (AlexaFluor 594-conjugated anti-rabbit antibody and fluorescein isothiocyanate-conjugated anti-mouse antibody (Invitrogen). The cells were visualized for WN/DENV-2 and DCP1a staining using a Nikon Eclipse fluorescence microscope.For Western blot analysis, normal, replicon expressing and virus infected cell lysates were electrophoresed on 12% Nu-PAGE gels (Invitrogen) and electroblotted onto polyvinylidene difluoride membranes . Following a blocking step with Tris-buffered saline containing 0.1% Tween-20 and 5% dry milk, the membranes were incubated with a GW182 (TNRC6A) antibody (Abcam), followed by horseradish peroxidase-conjugated secondary antibody (Peirce). Bound horseradish peroxidase was visualized with an ECL substrate kit (Thermo Scientific). Membranes were stripped and reprobed with a rabbit anti \u03b2-actin antibody as a loading control.Total RNA was isolated from HeLa cells and stable HeLa expressing WNV replicon or from WNV infected ES cell supernatants using RNeasy kit (Qiagen). Reverse transcription was performed using superscript III first-strand synthesis superMix (Invitrogen). Quantitative real-time PCR was performed using a SYBR green PCR master Mix (Applied Biosystems) on 7900HT fast real-time PCR system (applied Biosystems). Amplification conditions were as follows: 95\u00b0C for 10 min, followed by 40 cycles of 95\u00b0C for 15 s, and 60\u00b0C for 1 min. The forward and reverse primers to amplify GAPDH were atggggaaggtgaaggtcg and gggtcattgatggcaacaatatc, and that for TNRC6 A, B and C isoforms were gaaatgctctggtccgctaca & atctcctcttcactggcaaactca; ggagcaaaagcacaccacctg & tgctcctgtatcatccatctcg; cccgccgcacctgtctct & ctgctgctctttggtctgc, respectively and for WNV were atcgccggacttatgttcg & ctttcgctagagcctgtgattt. Relative TNRC6 mRNA expression was normalized with GAPDH mRNA and calculated using the \u03b4Ct method.Student's t test was used to compare two groups of independent samples."} +{"text": "Feature-tracking (FT) analysis offers a novel, fast and practicable method to calculate strain from routinely acquired steady state free precession (SSFP) images without the need to perform additional tagged sequences. There is no validation of this technique, however, against a reference standard myocardial tagging analysis for any strain parameter other than mid-left ventricular whole slice circumferential strain in children. In an adult study of patients with dilated cardiomyopathy (DCM) and healthy controls, we sought to validate the FT method against spatial modulation of magnetization (SPAMM) tissue tagging analysis (Cardiac Image Modeling Package (CIMTAG), University of Auckland) for the computation of long axis function.We compared measures of peak systolic longitudinal strain from the horizontal long axis view using the 2 techniques in 30 patients . Normal healthy adults were identified from an ongoing prospective, observational research study examining the effects of living kidney donation on cardiovascular structure and function (NCT01028703) while DCM patients underwent routine tagging during CMR studies performed for clinical requirements. A timed retrospective off-line analysis was performed on matched tagged and SSFP slices by 2 independent blinded observers (WEM and RJT).Mean peak systolic longitudinal strain and strain rate determined by FT were significantly reduced in DCM patients compared with healthy controls . The mean peak systolic longitudinal strain of DCM patients determined by CIMTAG (-6.7 \u00b1 1.7%) and FT (-7.0 \u00b1 1.4%) was not significantly different (P = NS). In an analysis of all patients, mean FT systolic longitudinal strain values were highly correlated with CIMTAG values, with a Pearson correlation coefficient of 0.67 (P=0.0001, Figure This FT based assessment of longitudinal strain correlated highly with values derived from tagged images in a population with a wide range of left ventricular function. Furthermore, FT can be performed without the need for additional imaging and lengthy post-processing.WEM is funded by a BHF Clinical Research Fellowship"} +{"text": "Diffuse myocardial fibrosis may be a fundamental features of adverse myocardial remodeling in idiopathic non-ischemic cardiomyopathy. As T1-weighted cardiac magnetic resonance (CMR) imaging provides an alternative method of diffuse fibrosis quantification, we sought to assess the association of myocardial T1 value to left ventricular reverse remodeling (LVRR).2 at 24 months. A multivariable logistic regression analysis was performed to identify associations with LVRR.We performed CMR in 24 patients with idiopathic non-ischemic cardiomyopathy and also in 12 healthy volunteers as control subjects. T1 mapping was performed with post-contrast Look-Locker gradient echo. Baseline echocardiography as well as hemodynamic and metabolic data were collected at the time of CMR. Patients were followed over a median time of 8 months for LVRR which was defined as a left ventricular ejection fraction (LVEF) increase of \u226510 U and a decrease in indexed left ventricular end-diastolic diameter (LVEDD) of \u226510% or indexed LVEDD of < 33 mm/mLVRR was found in 8 patients (33%). Mean T1 value was substantially lower in patients without LVRR (240+26) compared to patients with LVRR and healthy controls (FigurePost contrast T1 value is a predictor of LV reversed remodeling in patients with idiopathic non-ischemic cardiomyopathy, independent of baseline LVEF and the presence of myocardial scar.None."} +{"text": "Here we investigated whether real and sham acupuncture were associated with differential changes in MOR BP in FM patients that were naive to acupuncture as compared to conditioned patients.Previous studies suggest acupuncture analgesic effects involve the release of endogenous opioid peptides. We have previously shown that while both acupuncture and sham reduced clinical pain in fibromyalgia (FM), only acupuncture increased mu-opioid receptor (MOR) baseline receptor availability 20 female FM patients (mean (SD) age 44.3 (13.6) years) were randomized to acupuncture (n=10) or sham (n=10) treatments over four weeks. Acupuncture involved insertion of nine needles into the body whereas sham did not penetrate the skin. Positron emission tomography (PET) with the mu-opioid radiotracer [11C] carfentanil was performed during the first and ninth treatments. Each session included a \u2018baseline\u2019 scan prior to needle insertion and \u2018treatment\u2019 scan during needle insertion. We compared the difference in MOR BP between baseline and treatment, before and after 4 weeks of acupuncture versus sham. MOR quantification performed with Logan plots resulting in voxel-by-voxel maps of MOR BP. Images were analyzed with SPM 5.Real and sham acupuncture differed in their acute effects on MOR BP. The effect was localized in the left temporal pole and left amygdala/striatum . The treatment effect was largely due to a change in BP with sham after 4 weeks.Acupuncture and sham resulted in differential acute effects on MOR binding in treatment naive as compared to conditioned patients. These findings could demonstrate a conditioned placebo response in sham treated chronic pain patients."} +{"text": "Hypercapnic acidosis is often seen in critically ill patients and during protective mechanical ventilation. Conflicting findings regarding the effect of hypercapnic acidosis on endogenous nitric oxide (NO) production and hypoxic pulmonary vasoconstriction (HPV) have been reported. The aim of this study was to test the effects of hypercapnic acidosis on HPV, and the endogenous NO production in hypoxic and hyperoxic lung regions.2 inhalation (Hypercapnia group) to both lung regions, and eight pigs served as the Control group. The NO concentration in exhaled air (ENO), nitric oxide synthase (NOS) activity in lung tissue, and regional pulmonary blood flow were measured.Sixteen healthy anesthetized pigs were separately ventilated with hypoxic gas to the left lower lobe (LLL) and hyperoxic gas to the rest of the lung. The pigs were then randomized into two groups. Eight pigs received 10% CO2+-independent, or Ca2+-dependent NOS activity in hypoxic or hyperoxic lung regions. The relative perfusion to the hypoxic LLL (QLLL/QT) increased during the first 90 minutes of hypercapnia from 6 (1)% (mean (SD)) to 9 (2)% (P < 0.01), and then decreased to the same level as in the Control group where QLLL/QT remained unchanged over time (P > 0.05). In addition, hypercapnia increased cardiac output (QT) (P < 0.01), resulting in increased oxygen delivery (P < 0.01), despite a significant decrease in PaO2 (P < 0.01).There were no significant differences between the Hypercapnia and Control groups for ENO, CaHypercapnic acidosis does not affect the endogenous pulmonary NO production, nor does it potentiate HPV."} +{"text": "In previous studies using candidate gene approaches, low sperm count (oligospermia) has been associated with altered sperm mRNA content and DNA methylation in both imprinted and non-imprinted genes. We performed a genome-wide analysis of sperm DNA methylation and mRNA content to test for associations with sperm function.P\u200a=\u200a0.01). Linear models of microarray analysis (LIMMA) was performed based on motility and identified 9,189 CpG loci with significantly altered methylation (Q<0.05) in the low motility samples. In addition, the majority of these disrupted CpG loci (80%) were hypomethylated. Of the aberrantly methylated CpGs, 194 were associated with imprinted genes and were almost equally distributed into hypermethylated and hypomethylated groups. Sperm mRNA was measured with the Human Gene 1.0 ST Affymetrix GeneChip Array. LIMMA analysis identified 20 candidate transcripts as differentially present in low motility sperm, including HDAC1 (NCBI 3065), SIRT3 (NCBI 23410), and DNMT3A (NCBI 1788). There was a trend among altered expression of these epigenetic regulatory genes and RPMM DNA methylation class.Sperm DNA and mRNA were isolated from 21 men with a range of semen parameters presenting to a tertiary male reproductive health clinic. DNA methylation was measured with the Illumina Infinium array at 27,578 CpG loci. Unsupervised clustering of methylation data differentiated the 21 sperm samples by their motility values. Recursively partitioned mixture modeling (RPMM) of methylation data resulted in four distinct methylation profiles that were significantly associated with sperm motility (Using integrative genome-wide approaches we identified CpG methylation profiles and mRNA alterations associated with low sperm motility. Traditional semen analysis measures sperm concentration, motility, morphology, and semen volume, and is acknowledged to be a poor predictor of fertility, demonstrating remarkable intra- and inter-individual variability DNA methylation is the stable, covalent addition of a methyl group to cytosine that can represent response to environmental cues or exposures that may modify gene expression. Both human and animal studies indicate that abnormal sperm DNA methylation patterns are associated with subfertility, including aberrant methylation of both imprinted In addition to DNA methylation, significant effort is being devoted to developing human sperm mRNAs as biomarkers of infertility In the present study, we utilized high-density array techniques to investigate the hypothesis that alterations to the pattern of sperm DNA methylation or mRNA content are associated with sperm function.The Committee on the Protection of Human Subjects: Rhode Island Hospital Institutional Review Board 2 (Committee #403908) approved the study and written informed consent was obtained from all participants. Clinical investigation was conducted according to the principles expressed in the Declaration of Helsinki.et al., 2002) as detailed in the MGED Society website http://www.mged.org/Workgroups/MAIME/maime.html. This data is accessible through GEO Series accession number GSE26982 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE26982).The microarray data discussed in this publication is MAIME compliant and the raw data has been deposited in NCBI's Gene Expression Omnibus gradient to eliminate debris, non-sperm cells, and dead sperm Prior to processing the 21 samples, sperm purity was confirmed by the absence of somatic cell contaminants using bright phase microscopy and by the absence of 18/28S ribosomal RNA peaks by RNA gel electrophoresis (data not shown) www.Illumina.com). Multiple groups including ours have previously demonstrated the validity of Illumina methylation array data using several different approaches DNA was isolated from the sperm of the 21 men using a modified protocol in which sperm pellets were lysed for 16 hours in a solution containing Tris , DTT , NaCl , EDTA , SDS , Proteinase K , and beta-mercaptoethanol http://www.geneimprint.com/databases/ and http://igc.otago.ac.nz/home.html) (n\u200a=\u200a62); (2) imprinted genes identified using the ChIP-SNP method (n\u200a=\u200a27) A list of 187 imprinted genes in the human genome was compiled based on information from three sources: (1) experimentally determined imprinted genes listed in two databases protocol in addition to components of Qiagen's RNeasy kit . Using the Brown Genomics Core Facility, the isolated sperm mRNA was processed and hybridized to Affymetrix GeneChip Human Gene 1.0 ST Arrays , providing whole-transcript coverage of 28,869 genes by \u223c26 probes spread across the length of each gene. The probe cell intensity data from the Affymetrix GeneChips was normalized and annotated using Affymetrix Expression Console as recommended by the manufacturer. The application uses the RMA-Sketch workflow analysis as the default to create CHP files. The CHP log2 expression files were then merged in Expression Console with the annotation file and the annotated log2 results were exported as a text file for third-party downstream analysis.Aside from array normalization procedures, the R software environment was used for all statistical analysis.Recursively partitioned mixture modeling (RPMM) profiles were fit to the entire Infinium array using previously described methods The LIMMA procedure a priori hypothesis for association with subfertility existed based on previous reports. The analysis included 177 imprinted genes as well as 99 candidate genes with biallelic expression with few (\u223c400) consistently expressed in sperm pression [10], [1Associations among the RPMM classes and the normalized gene expression values for candidate transcripts were calculated with the KW test statistic utilizing the strategy employed previously. Messenger RNAs were considered significantly associated with RPMM class when P<0.02, after adjusting for multiple comparisons using the Bonferroni correction.Unsupervised clustering of sperm DNA methylation data for the 1,000 most variable CpG loci on the array highlights the methylation differences among the 21 individual men . As showP\u200a=\u200a0.01). The association between RPMM methylation class and sperm morphology approached statistical significance (P\u200a=\u200a0.09), though methylation class was not associated with sperm count (P\u200a=\u200a0.29).Recursively partitioned mixture modeling (RPMM) was performed on raw methylation data to organize the sperm samples into methylation classes based on similarity. The algorithm first separated the 21 sperm profiles into two different branches left (L) and right (R) and then further subdivided each branch into right and left branches resulting in 4 total classes: left left (LL), left right (LR), right left (RL) and right right (RR) . In FiguQ<0.05) was used to univariately test each CpG for association with motility. 9,189 of 27,578 CpGs (34%) had significantly altered methylation associated with motility after adjusting for multiple comparisons (Q<0.05) . Of thesBecause establishing proper methylation marks within imprinted genes during spermatogenesis is critical, we next restricted our analysis to CpGs associated with imprinted genes. Of the 616 CpGs associated with imprinted genes, 194 CpGs (31.5%) had significant associations with motility, similar to the distribution of the array overall. Amongst these loci, 47% (n\u200a=\u200a92) were hypermethylated in the low motility samples, whereas 53% (n\u200a=\u200a102) were hypomethylated. The majority of hypomethylated CpGs were on maternally expressed genes (45%), followed by paternally expressed (33%) and those with undetermined parent of expression (22%). Conversely, the majority of hypermethylated CpGs were associated with paternally expressed genes (70%), with the remainder maternally expressed (26%), and of undetermined parental expression (4%). The 194 loci corresponded to 92 genes, with 11 genes showing both hyper- and hypomethylated loci .Aberrant promoter methylation in genes related to spermatogenesis and epigenetic regulation have recently been identified in sperm from men with poor semen quality Q<0.05) (GLI3 (NCBI 2737), APAB1 (NCBI 320), CTNND2 (NCBI 1501), FERMT2 (NCBI 10979), PHPT1 (NCBI 29085), SNRPN (NCBI 6638), PPP1R9A (NCBI 55607), CDH18 (NCBI 603019), ALDH1L1 (NCBI 10840), LDB1 (NCBI 8861), and PEX10 (NCBI 5192)), six genes associated with spermatogenesis (SERPINA5 (NCBI 5104), ACE (NCBI 1636), FANCC (NCBI 2176), PCSK4 (NCBI 57460), CYP19A1 (NCBI 1588), and FAS (NCBI 355)), and three epigenetic regulatory genes . HDAC1, DNMT3A, LDB1 and FAS showed increased mRNA content in the low motility samples, whereas the remaining 16 showed decreased mRNA content.Focusing on imprinted mRNAs and candidate biallelic mRNAs, LIMMA analysis was performed to identify differentially expressed transcripts, conditioning on motility. Twenty genes were identified as significant after adjusting for false discovery rate (Q<0.05) . These iHDAC1, SIRT3, and DNMT3A) with significantly altered expression in low motility sperm . For all three genes, the association between mRNA expression level and methylation class membership approached significance after adjusting for multiple comparisons .It is known that major modifications in chromatin organization occur in spermatid nuclei during spermatogenesis, leading to the high degree of packaging in the sperm head. Chromatin compaction ensues when the histones surrounding the DNA are replaced by protamines, and this occurs in parallel with transcriptional arrest ty sperm . Among mCurrently, the evaluation of male infertility relies upon physical exam and semen and hormone analyses; although quick and relatively inexpensive, these physiologic measurements often do not explain the underlying cause of infertility nor predict the usefulness of various therapeutic interventions. Therefore, new approaches are needed to identify the etiologies of male infertility. Recent data suggest that sperm DNA methylation abnormalities and alterations in sperm mRNA content are found in infertile men Due to the unreliable nature of classifying men into abnormal and normal groups during a semen analysis, we used a data driven approach to first qualitatively assess associations among sperm DNA methylation and our patient population. Unsupervised clustering indicated that there was an association between DNA methylation and motility status. This was true both for all of the CpGs on the array and the imprint-only subset.RPMM separated the 21 men into four classes based on similarity of DNA methylation array data. The median motility values were calculated for each class and the results suggested that the methylation profiles were associated with motility. Comparing the DNA methylation heatmap to the class versus motility boxplot indicates that the low motility class has the most aberrantly methylated CpGs. Overall, these data suggest that low motility sperm have increased hypomethylation relative to high motility sperm. We used LIMMA to identify the significantly altered CpGs conditioned on changes in motility for all CpGs on the array: over one-third of the CpGs were significantly differentially methylated in the low motility samples and the majority of these were hypomethylated. The high prevalence of aberrantly methylated CpGs suggests a genome-wide DNA methylation defect in the low motility sperm. It has been previously hypothesized that the aberrant sperm DNA methylation could be due to abnormal chromatin compaction, inefficient DNA methyltransferases, and/or failure to maintain or acquire the correct methylation marks during spermatogenesis and our results are consistent with this literature DIRAS3 (NCBI 9077), H19 (NCBI 283120), IGF2 (NCBI 3481), MEST/PEG1 (NCBI 4232), PLAGL1/ZAC (NCBI 5325), MEG3/GTL2 (NCBI 55384), and SNRPN) have already been noted as aberrantly methylated in abnormal sperm PEG3 (NCBI 5178) and LIT1/KCNQ1OT1 (NCBI 10984)) has been inconsistently reported in the literature We initially focused on CpGs mapping to imprinted genes because of their plasticity during spermatogenesis, biological relevance following conception and development, and because previous studies have identified imprinted loci as aberrantly methylated in abnormal sperm a priori hypothesis for association with male subfertility or epigenetic regulation exists. Twenty genes were identified as demonstrating significantly altered transcript levels in low motility sperm. All of the mRNAs except HDAC1, DNMT3A, LBD1, and FAS were present in decreased amounts in low motility sperm, and we did not observe altered mRNA content for BRDT, which was previously reported to have increased expression in subfertile patients To further clarify the potential functional alterations to imprinted genes and critical epigenetic regulatory genes, we evaluated sperm mRNA content of 177 imprinted genes and 99 other transcripts where an HDAC1, SIRT3, and DNMT3A) and methylation class. HDAC1 is the predominant histone deacetylase (HDAC) during spermatogenesis. Histone hyperacetlyation is required for the histone to protamine exchange and is facilitated by the degradation of HDAC1 in elongated spermatids Integration of epigenetic and expression data revealed a relationship between transcript content of three epigenetic regulatory genes low motility sperm have reduced SIRT3 mRNA content which might be related to increased subcellular ROS during spermatogenesis leading to the abnormal motility phenotype; and (3) this oxidative stress may be impeding the ability of DNMT3A to set the correct methylation marks which would also contribute to the hypomethylated phenotype. Our results suggest that additional integrative studies including larger sample sizes as well as prospective studies of fertility following these integrated molecular assessments have great potential to advance our understanding of the molecular features of sperm associated with fertility status.Although we were limited by sample size, we used a powerful integrative approach to simultaneously examine sperm DNA methylation and mRNA content utilizing two high density array techniques. We found that: (1) low motility sperm have genome-wide DNA hypomethylation that may be due to a failure of the sperm to complete chromatin compaction properly because of increased Table S1Imprinted Genes.(DOC)Click here for additional data file.Table S2Genes Associated with Spermatogenesis and Epigenetic Regulation.(DOC)Click here for additional data file.Table S3Aberrant CpGs in Low Motility Sperm.(DOC)Click here for additional data file.Table S4Aberrant mRNA Transcripts in Low Motility Sperm.(DOC)Click here for additional data file."} +{"text": "Current clinical practice widely regards deep tissue biopsy as the gold standard for identification of wound bacterial bio-burden. This study aims to establish whether fine needle aspiration biopsy (FNAB) is as accurate as deep tissue biopsy and therefore offers a more accurate, cheaper and suitable alternative to routinely used superficial swab in diabetic wounds of varying depth and severity.A total of 15 infected diabetic foot wounds were sampled and cultured. Three specimens were taken from each wound: superficial swab before debridement, followed by fine needle aspiration biopsy and deep tissue specimen at the end of sharp debridement.FNAB identified the same significant bacteria found in deep tissue culture in only 5 (33%) wounds missing all or some significant bacterial isolates in 10 (67%) wounds. In 6 (40%) of these wounds FNAB cultures were negative. In comparison swab cultures identified the same significant bacterial isolates found in deep tissue culture in 13 (87%) wounds, missing 1 bacterial isolate in 2 (13%) wounds with no negative cultures recorded.In this limited sample it would appear that FNAB culture technique is severely inadequate in identifying pathogens in diabetes foot wounds in comparison to superficial swab technique and gold standard deep tissue biopsy. FNAB cultures missed microorganisms in two thirds of wounds and had a high false negative rate."} +{"text": "TLR4 polymorphisms withsusceptibility/resistance to malaria disease. In the current immunogenetic study, weassessed the TLR4 genotypes formed by the two common single nucleotide polymorphisms(SNPs) (Asp299Gly and Thr399Ile) in the co-segregate state in BaluchiPlasmodium falciparum infected and healthy populations from malaria hypoendemicareas of Iran. The study was performed to evaluate the distribution and correlation ofTLR4 co-segregating genotypes in patients with mild malaria. Moreover, the frequencyof these genotypes was compared with reported results from other populations insimilar or contrasting malaria settings around the world.Different studies have shown an association of TLR4gene (Asp299Gly and Thr399Ile) were analyzed in 350 Baluchi patients with mild malariaand 350 unrelated healthy controls by using polymerase chain reaction/restriction fragmentlength polymorphism (PCR/RFLP) techniques followed by sequencing analysis. Differencesin the TLR4 co-segregate genotype frequencies among the studied group weredetermined by Fisher\u2019s exact test.In this case control study, the presence of 2 SNPs in the TLR4 genotypespresented a diverse and distinct pattern in the Baluchi population, no significant differencewas detected between the cases and controls (p>0.05). A lower frequency of TLR4Asp299Gly/Thr399Thr was observed in Baluchis with mild malaria compared to Africanpopulations (p< 0.05).Although the distribution of the two commonly co-segregating TLR4 Asp299Gly/Thr399Ilegenotypes in the Baluchi population compared to other malaria endemic populations maysuggest different local evolutionary pressure on TLR4 polymorphisms by malaria in thisregion. The higher frequency of Asp299Gly/Thr399Ile genotypes among the Baluchi populationcompared with the African population (p< 0.05) which suffers from a larger numberof severe cases might suggest that this genotype has a role in protecting against severemalaria. These findings are useful for further understanding the pathogenesis of severemalaria.Differences in the co-segregation patterns of Plasmodium-infected individuals develop severeand complicated symptoms, while others remainasymptomatic or develop mild malaria.Malaria is one of the most important infectiousdiseases and every year 350-500 million cases ofmalaria are reported worldwide . The cliPlasmodium infection thatreleases inflammatory cytokines to clear the parasitefrom the circulation but may also contributeto disease severity receptor and Thr399Ile (rs4986791) that arelocated in the leucine-rich repeat domain responsiblefor ligand recognition. These mutations affectthe ligand-binding region (Asp299Gly) of TLR4and the co-receptor-binding region (Thr399Ile)of the receptor in the co-segregate state in mild malariapatients and healthy individuals in the Baluchipopulation from Iran where there has beenno report of severe malaria cases. In addition, the frequency of the common TLR4 co-segregategenotypes was compared between the Baluchipopulation who are living in malaria hypoendemicareas of Iran with those reported results from otherpopulations from similar or contrasting malariasettings around the world. These results are used toobtain an insight into the evolutionary pressure ofinfections, particularly malaria on the TLR4 polymorphismsin a population from the Middle East.In the present investigation, which is a continuationof our previous work , we analThe malaria endemic regions of Iran are situatedin the south-eastern part of the country including theprovinces of Sistan and Baluchistan, Hormozganand Kerman. In Iran, the incidence of malaria has declinedgradually over the last few years from 15,712in 2007 to 3,015 cases in 2010 due to the beginningof elimination strategies . In these regions, there are no reports of anaemia,severe malaria or death due to malaria and mostpatients are adults with mild malaria. This study wascarried out in the Chabahar District of the Sistan andBaluchistan Province in south-eastern Iran.In this case-control study, the population-basedcontrols were selected from healthy Baluchi individualswith no exposure or last exposure to malariaparasite during the past 10 years. Exposure statuswas obtained by interviewing the participant/orspouse or other family member, as well as searchingthe participant\u2019s medical records in the malariahealth center of the study area over the last 10 years.P. falciparum were recruitedfrom outpatient clinics at primary health centers inChabahar district during 2003-2009. These patients,who presented with fever (in the preceding 48 hourswith an axillary temperature \u226537.5\u02daC), or muscularpain and headache, were considered symptomaticand classified as having mild malaria. In addition,all patients had mono-infection with P. falciparum,previous history of malaria and parasitaemia rangingbetween 1,000 and 35,000 asexual parasites/mm3.The P. falciparum parasite species was diagnosed viamicroscopy locally in the study area and confirmedby nested PCR assay were typed in all 700 participants(case and control). These two SNPs were analyzedusing PCR-RFLP analysis. Primers sequencesand PCR conditions for these polymorphisms werereported in our previous study that assessed TLR4polymorphisms and also TLR9 and TIRAP polymorphismsin 640 Baluchi individuals and 0.2 U Taq polymerase. TLR4 Asp299Gly amplificationyielded a 213 base-pair (bp) fragment. ThePCR reaction consisted of one initial cycle at 95\u02daCfor 5 minutes, 35 cycles at 94\u02daC for 1 minute, at 57\u02daCfor 1 minute and 72\u02daC for 1 minute followed by 57\u02daCfor 2 minutes and 72\u02daC for 10 minutes. For TLR4Thr399Ile, amplification yielded a 185 bp fragment inwhich the PCR program consisted of one initial cycleat 95\u02daC for 5 minutes, 35 cycles at 94\u02daC for 1 minute,at 54\u02daC for 1 minute and 65\u02daC for 1 minute followedby 54\u02daC for 2 minutes and 65\u02daC for 10 minutes. ThePCR products were electrophoresed on a 2% agarosegel .In this study two non-synonymous SNPs of ividuals . BrieflyTLR4 Asp299Gly and Thr399IleSNPs, the PCR products were cleaved using Nco\u0406 andHinf\u0406 (Fermentas) restriction endonucleases, respectively.The mixture was incubated at 37\u02daC overnight,and then electrophoresed on a 3% agarose gel .To screen the Sequence analysis was carried out to evaluatethe results obtained by RFLP, using the primersdescribed previously . The PCRTLR4co-segregate genotype frequencies among thestudied groups were determined by Fisher\u2019sexact test using SPSS for windows (version16.0). A p value of < 0.05 was considered tobe significant. For comparison of the frequencyof TLR4 co-segregate genotypes that wereobserved in the Baluchi population with otherpopulations from different parts of the world,the chi-square analysis was used.The sample size was calculated using thesample sized-unmatched case control by meansof OpenEpi software . DiffereP. falciparumor P. vivax infections. All of the studiedparticipants were successfully assessed for TLR4Asp299Gly/Thr399Ile SNPs , Asp299Gly/Thr399Ile (9.4%), Asp299Asp/Thr399Ile (6%),Asp299Gly/Thr399Thr (2.6%) and Gly299Gly/Thr399Thr (1.4%). In non-infected participantsa similar distribution of genotypes; Asp299Asp/Thr399Thr (80.6%), Asp299Gly/Thr399Ile (6.6%),Asp299Asp/Thr399Ile (9.4%) and Asp299Gly/Thr399Thr (3.4%) was observed. Gly299Gly/Ile399Ile,Asp299Asp/Ile399Ile, Asp299Gly/Ile399Ileand Gly299Gly/Thr399Ile genotypes were not observedin the studied population.In the assessment of co-segregate TLR4(Asp299Asp/Thr399Thr) had the highest frequencyin both studied groups and the mutantgenotype (Gly299Gly/Thr399Thr) had the lowestfrequency in only Baluchi infected subjects was significantly lower in the Baluchiin comparison with the African and Asianpopulations in Baluchi patients with mildmalaria is reported for the first time. The findingsare in agreement with a recent report by Loana et al.. This resultsuggests that different TLR4 Asp299Gly/Thr399Ileco-segregating genotypes might not be associatedwith mild malaria in our studied population.In the assessment of the two TLR4 Asp299Gly/Thr399Thr heterozygouscells released significantly higher TNF-\u03b1 thanthe wild type TLR4 Asp299Asp/Thr399Thr. However,the double heterozygote genotype (Asp299Gly/Thr399Ile) responded in a similar way to the wildtype genotype (Asp299Asp/Thr399Thr). It has alsobeen shown that the presence of the TLR4 Gly299Gly/Thr399Thr genotype is associated with an increasein susceptibility to malaria but a reduction inmortality and cerebral malaria in the Ghanaian andCameroonian populations (TLR4 Gly299Gly/Thr399Thr (1.4%) among Baluchi infected individuals,its absence in healthy participants and thehigh frequency of Asp299Asp/Thr399Thr (80.6%)among the studied population from an area with noreport of severe malaria or mortality due to malariamight suggest an association of these genotypes withclinical symptoms of mild malaria. This hypothesisneeds to be supported by analyzing TLR4 polymorphismsin more samples from diverse malaria hypoendemicregions.The cytokine study of the profile in LPS-stimulatedwhole blood cultures by Ferwerda et al. revealedulations , 22. In TLR4 Asp299Gly/Thr399Thr (3.4%) genotype among the Baluchipopulation compared to some African populations(e.g. 9.4% in Sudanese inhabitants) may support a role for this genotype in susceptibility to severemalaria with alarge number of severe malaria cases among the Baluchipopulation than the African population . The significantly lower frequencies ofthis co-segregate genotype in the African population,which has a larger number of severe cases ofmalaria than the Baluchi population, suggests thatthe Asp299Gly/Thr399Ile genotype may help protectagainst severe malaria. However, functionalstudies are needed to draw final conclusions on theeffect of the Asp299Gly/Thr399Ile genotype and itsassociation with malaria severity in this population.One of the most notable findings of this studywas the higher frequency of the TLR4 Asp299Gly/Thr399Ile co-segregate genotypes werereported in the Baluchi population. Comparisonwith other malaria endemic populations suggestsdifferent local evolutionary pressures on TLR4 polymorphismsby malaria or other infectious diseasein this region. A significantly lower frequency ofthe Asp299Gly/Thr399Thr genotype in the Baluchipopulation in comparison with the Sudanese andDogon populations (p< 0.05), which have a largenumber of severe malaria cases, might support arole for this genotype in susceptibility to severe malaria.These findings need to be extended to populationsof different ethnicity from diverse malariaendemic regions for further understanding of thepathogenesis of this serious disease.In the present study, mixed patterns of"} +{"text": "We tested whether 23 HIV-1 epitopes designed using a bioinformatics approach were recognized by HIV-infected children living in ethnically diverse Cameroon, which harbors a high HIV strain variety.Areas containing both promiscuous HLA-class I and II epitopes covering more than 90% of the HLA haplotypes present in the African population were identified within the most HIV-1 conserved sequence of each protein. Twenty-three peptides have been designed targeting gag (10), nef (6), tat (4), vpr (2) and vpu (1) proteins.We enrolled 33 children vertically infected with HIV (age range: 3 months \u2013 12 years), na\u00efve for antiretroviral therapy, with CD4% >15% or CD4 count >350 cells/ml.T cell recognition of single peptides was assessed by IFN-\u03b3 ELISpot. Pooled peptides were used to assess helper (CD40L expression) and cytotoxic (CD107a expression on cell surface) functions, and proliferative capacity (CFSE dilution) by flow cytometry.The majority (76%) of children recognized at least one peptide or peptide pool in at least one test, but none of the functional assays alone would have identified all responders.Half (48%) of the peptides were recognized by at least one child in ELISpot assay, nef peptides being the most frequently targeted (by 67% of the responders). Three previously undescribed epitopes were identified, while 2 epitopes commonly considered immunodominant were not recognized.In some children the same peptides were able to elicit different functions, while in other children diverse functions were induced by different peptides. The breadth of the epitope recognition and the number of different functions elicited were directly correlated and independent from the length of infection (age).These data provide a proof-of-concept for the rational design of T cell immunogens by reverse immunogenetic approach and support the parallel use of different functional assays for epitope mapping."} +{"text": "Previous studies have shown that magnesium sulfate (MgSO4) administered in a patient with a diffuse axonal injury (DAI) can serve as a useful neuroprotective agent. The present study was conducted to examine whether magnesium sulfate has a therapeutic efficacy and safety in patients with a severe diffuse axonal injury. Adult patients admitted within 1 hour of a closed Traumatic Brain Injury (TBI) with a severe diffuse axonal injury that met eligibility criteria were randomly selected and divided into two groups. Our treatment guidelines consisted of an initial loading dose of 50 mg/kg magnesium sulfate and then 50 mg/kg QID (quarter in die) up to 24 hours after the trauma. The outcome measures were mortality, Glasgow Coma Scale (GCS) score, and motor function scores which were assessed up to 2 months post-trauma. Magnesium showed a significant positive effect on GCS score at 2 months post-trauma (P=0.03). Motor functioning scores of patients in the MgSO4 group were higher than those in the control group but this was not statistically significant .Findings of the present study demonstrated that administration of magnesium sulfate following severe DAI can have neuroprotective effect.Severe diffuse axonal injury, Magnesium sulfate, Outcome"} +{"text": "TRPA1 receptor channels are activated by environmental irritants and by endogenous mediators released during inflammatory conditions . ActivaTo further examine the role of TRPA1 receptor activation in processes presumably associated with headache generation, we investigated the effects of the TRPA1 agonist acrolein on functions involved in meningeal nociception using four different rat models.The discharge activity of single meningeal afferents innervating the dura mater was recorded in a hemisected cranial preparation. 2. The activity of second order neurons in the spinal trigeminal nucleus with meningeal afferent input was recorded in anaesthetised animals. 3. In the hemisected cranial preparation the dura mater was superfused with synthetic interstitial fluid, and stimulated calcitonin gene-related peptide (CGRP) release was measured using an ELISA. 4. Meningeal blood flow was monitored in the exposed dura mater of anaesthetised animals using laser Doppler flowmetry. In all preparations the dura mater was stimulated with the TRPA1 agonist acrolein (10-4 M).Acrolein did not elicit discharges in meningeal A\u03b4- or C-fibres in the hemisected cranial preparation and did not change the discharge activity of second order neurons with meningeal receptive fields in anaesthetized animals. In contrast, acrolein significantly stimulated CGRP release from the dura mater within 5 min and increased meningeal blood flow. Both responses were suppressed by the TRPA1 inhibitor HC030031.TRPA1 channel activation causes neuropeptide release from meningeal afferents but does not generate propagated afferent information. Therefore an important role for peripheral TRPA1 receptors in headache generation appears unlikely."} +{"text": "We sought to assess late gadolinium enhancement (LGE) of the coronary artery wall and ascending aorta in patients with Takayasu arteritis (TA) compared to patients with stable CAD. Our findings suggest that LGE of the coronary artery wall can be demonstrated in patients with TA and seems to be similarly pronounced as in CAD patients. The observed coronary LGE seems rather unspecific and differentiation between coronary vessel wall fibrosis and inflammation still remains unclear.Takayasu arteritis (TA) is a rare chronic inflammatory granulomatous arteritis of the aorta and its major branches. Late gadolinium enhancement (LGE) with magnetic resonance imaging (MRI) has demonstrated its value for the detection of vessel wall alterations of large arteries in TA ). InvolvWe enrolled 9 patients fulfilling the ACR criteria of TA wiNo coronary vessel wall enhancement was observed prior to contrast in either group. Post contrast, LGE on IR scans was detected in 28 of 50 segments (56%) seen on T2Prep in TA and in 25 of 57 segments (44%) in CAD patients. Quantitative LGE analysis of the ascending aorta showed a significant increase of contrast-to-noise-ratio (CNR) between pre and post contrast agent administration in TA (5.3\u00b14.5 vs. 13.5\u00b15.7) and in CAD patients (2.5\u00b12.3 vs. 4.7\u00b12.4), with p=0.028 for both groups. A highly significant difference for CNR of the aortic wall post contrast between TA and CAD patients was observed (p=0.001). LGE quantitative assessment of coronary artery vessel wall CNR post contrast revealed no significant difference between the two groups .Our findings suggest that LGE of the coronary artery wall can be demonstrated in patients with TA and seems to be similarly pronounced as in CAD patients. The observed coronary LGE seems rather unspecific and differentiation between coronary vessel wall fibrosis and inflammation still remains unclear.none."} +{"text": "During human walking the ankle-foot complex executes seemingly contradictory functions: 1) stabilization of the human body at initial contact, (2) shock absorption during early stance stabiliz, (3) StoUsing a seven-camera motion capture system (250Hz), a pressure platform (500Hz) and a forceplate (1250Hz), we measured forefoot deformation through kinematic and pressure related outcome measures in 60 healthy subjects.Small but significant changes in intermetatarsal distance are established during stance phase, with the largest change occurring between metatarsal head II/III and V Table . The chaHigh correlation values (>0.7 or <-0.7) are found between temporal pressure and temporal kinematic parameters.Through stance the forefoot deforms according to a specific pattern, which is predominantly determined through forefoot-ground interaction. In addition, the changes in forefoot kinematics in combination with temporal contact data argue the existence of a mediolateral metatarsal arch and suggest the existence of an inverse arch during metatarsal forming and final propulsion phase."} +{"text": "To systematically review evidence on depression screening in coronary heart disease (CHD) by assessing the (1) accuracy of screening tools; (2) effectiveness of treatment; and (3) effect of screening on depression outcomes.A 2008 American Heart Association (AHA) Science Advisory recommended routine depression screening in CHD.CINAHL, Cochrane, EMBASE, ISI, MEDLINE, PsycINFO and SCOPUS databases searched through December 2, 2011; manual journal searches; reference lists; citation tracking; trial registries. Included articles (1) compared a depression screening instrument to a depression diagnosis; (2) compared depression treatment to placebo or usual care in a randomized controlled trial (RCT); or (3) assessed the effect of screening on depression outcomes in a RCT.There were few examples of screening tools with good sensitivity and specificity using a priori-defined cutoffs in more than one patient sample among 15 screening accuracy studies. Depression treatment with antidepressants or psychotherapy generated modest symptom reductions among post-myocardial infarction (post-MI) and stable CHD patients , but antidepressants did not improve symptoms more than placebo in 2 heart failure (HF) trials. Depression treatment did not improve cardiac outcomes. No RCTs investigated the effects of screening on depression outcomes.There is evidence that treatment of depression results in modest improvement in depressive symptoms in post-MI and stable CHD patients, although not in HF patients. There is still no evidence that routine screening for depression improves depression or cardiac outcomes. The AHA Science Advisory on depression screening should be revised to reflect this lack of evidence. Major depressive disorder (MDD) is present in approximately 20% of coronary heart disease (CHD) patients The AHA invests considerable resources in ensuring that practice guidelines are revised rapidly to reflect new evidence Key Question #1: What is the accuracy of depression screening instruments in CHD?Key Question #2: Does treatment of depression in CHD improve depressive symptoms or cardiac outcomes?Key Question #3: Does depression screening in CHD improve depression outcomes?This systematic review updates a previous review from November 2008 To update the previous review Diagnostic and Statistical Manual of Mental Disorders diagnosis of MDD or an International Classification of Diseases depressive episode, established with a validated diagnostic interview administered within 2 weeks of the screening tool. Eligible articles for Key Question #2 were RCTs comparing depression treatment with placebo or usual care among CHD patients with MDD or an International Classification of Diseases depressive episode based on a validated diagnostic interview. For trials of patients with MDD and other conditions , we sought original study data for patients with MDD for trials with 80% power to detect a 0.50 standardized mean difference effect size (n\u200a=\u200a64 per group). Eligible articles for Key Question #3 were RCTs that compared depression outcomes between CHD patients who underwent depression screening and those who did not.Eligible articles were original studies in any language with data on adult patients in cardiovascular care settings based on diagnosis or procedure, including mixed populations if CHD data were reported separately. Eligible diagnostic accuracy studies (Key Question #1) reported data allowing determination of sensitivity, specificity, positive predictive value, and negative predictive value compared to a Two investigators independently reviewed titles/abstracts for eligibility with full-text review of articles identified as potentially eligible by one or both. Disagreements after full-text review were resolved by consensus. Chance-corrected agreement was assessed with Cohen's \u03ba.Two investigators independently extracted study data (File S4) and assessed risk of bias with discrepancies resolved by consensus. Risk of bias was assessed with the revised Quality Assessment for Diagnostic Accuracy Studies tool for Key Question #1 (File S5) and the Cochrane Risk of Bias tool for Key Question #2 (File S6).g statistic (standardized difference between 2 means). The most comprehensive cardiovascular outcome available was extracted.For Key Question #1 (diagnostic accuracy), data were extracted based on optimal cutoffs identified by study authors. For Key Question #2 (treatment) when multiple outcomes were reported, designated primary outcomes were prioritized, followed by observer-rated scales, then self-report measures. Post-intervention effect sizes were reported using the Hedges's For Key Question #1, studies were heterogeneous in terms of patient samples, screening tools and cutoffs, and whether they used standard scoring thresholds versus sample-specific thresholds based on exploratory data analysis. For Key Question #2, studies had heterogeneous patient samples, therapeutic interventions, and treatment durations. No eligible studies were identified for Key Question #3. Thus, results were not pooled quantitatively.Of 1,442 citations, 1,405 were excluded after title/abstract review and 29 after full-text review, leaving 8 eligible articles ; \u03ba\u200a=\u200a1 , 0.07 to 0.33). This was compared to usual care. Among depressed patients enrolled in the ENRICHD trial, 28% of patients in the cognitive behavior therapy intervention arm and 21% in the usual care arm were prescribed an antidepressant in the 12 months following trial enrolment. The other psychotherapy trial, the Canadian Cardiac Randomized Evaluation of Antidepressant and Psychotherapy Efficacy trial There were 2 studies where psychotherapy was investigated. The Enhancing Recovery in Coronary Heart Disease Patients (ENRICHD) trial No studies reported improved cardiac outcomes, although only ENRICHD Risk of bias ratings are shown in File S8 for each treatment study. Three trials were rated low risk of bias for most categories Of 1,453 unique titles/abstracts, 1 received full-text review, and no eligible RCTs were identified . PotentiThe main findings of this systematic review are that: (1) there are few examples of screening tools with high sensitivity and specificity using an a priori-defined cutoff score in more than one CHD sample. When results from studies that used a pre-specified score were available in more than one sample, estimates of diagnostic accuracy were inconsistent. When exploratory data analysis methods were used to both generate a cutoff score and assess the accuracy of that cutoff score in the same sample, different studies tended to produce cutoffs that were inconsistent across studies; (2) depression treatment improves symptoms of depression in post-MI and stable CHD patients, although symptom reductions are modest; (3) antidepressant treatment has not reduced depressive symptoms compared to placebo in two trials with HF patients, although one small trial was stopped prematurely, and the other trial provided nurse-facilitated depression management services to patients in both the antidepressant and placebo groups; and (4) no RCTs have evaluated whether routine depression screening in CHD would improve depression outcomes.The AHA has recommended that all CHD patients be routinely screened for depression In cardiovascular care settings, several observational studies have reported on the administration of depression questionnaires. One study In primary care, the UK's Quality and Outcomes Framework pay-for-performance program introduced routine depression screening for patients with CHD or diabetes in April 2006 The AHA website lists more than 20 patient care guidelines issued since the 2008 AHA Advisory on depression screening Depression screening can benefit patients only to the extent that it identifies depressed patients not already diagnosed or treated for depression, successfully enrolls those patients in treatment, and achieves positive treatment results. As described recently Without evidence that depression screening benefits CHD patients, the potentially considerable resources and costs that would be involved in implementing routine screening must be even more carefully considered Depression screening would almost certainly increase the number of patients using antidepressants, and potential harms of even more widespread use of antidepressants by CHD patients must therefore be considered. Selective serotonin reuptake inhibitors (SSRIs) may act as antiplatelet agents, which can increase the risk of major bleeding, especially when used along with the combination of aspirin and a thienopyridine antiplatelet drug like clopidogrel The AHA recommendation for depression screening in CHD File S1Search Strategies.(DOC)Click here for additional data file.File S2Relevant Systematic Reviews.(DOCX)Click here for additional data file.File S3Journals Included in Manual Searching.(DOCX)Click here for additional data file.File S4Variables Included in Data Extraction Forms.(DOCX)Click here for additional data file.File S5QUADAS-2 Risk of Bias and Applicability Judgments.(DOCX)Click here for additional data file.File S6Cochrane Risk of Bias Tool Domains.(DOCX)Click here for additional data file.File S7Quality Assessment of Studies of Diagnostic Accuracy (QUADAS-2).(DOCX)Click here for additional data file.File S8Assessment of Risk of Bias in Randomized Controlled Trials in Key Question #2 (Treatment).(DOCX)Click here for additional data file.File S9Excluded Studies for Effect of Screening on Depression Outcomes (Key Question #3).(DOCX)Click here for additional data file."} +{"text": "Covalent modifications to cytosine provide important epigenetic information required for normal embryo development. 5\u2019methylcytosine (5mC) has received most experimental attention while 5\u2019hydroxymethyl cytosine (5hmC) has assumed recent prominence. Extant gold-standard methods of analysis of do not discriminate between 5mC and 5hmC hence most published methylomes have limits to their interpretation. Immunolocalization allows discrimination between the range of modifications, provides a genome-wide level of analysis, allows differential assessment of nuclear localisation, and is compatible with the limited DNA available within the early embryo. Using newly established methodology for full retrieval of these two antigens we have"} +{"text": "Tg737, (encoding IFT88) which abolishes cilia growth. 3D agarose culture allowed compressive loading of WT and Tg737 chondrocytes followed by quantification of ATP release with a luciferase assay, calcium transients by Fluo-4 imaging, and matrix synthesis by qPCR and biochemical assay. Additionally, expression of purinergic receptors (P2R) and polycystins was assessed by western blot and immunocytochemistry. Compression of WT chondrocytes increased calcium transients and matrix production. By contrast this mechanosensitive behaviour was completely abolished in Tg737 cells. However mechanosensitive ATP release was present in both WT and Tg737 cells suggesting that IFT88 and the cilium are required for purinergic reception. Indeed exogenous ATP elicited calcium transients in WT but not in Tg737 cells. P2R expression profiles showed no global differences but polycystin-1 expression was altered in ORPK. We conclude that IFT88 plays a critical role in ATP-induced calcium signalling and is therefore essential to chondrocyte mechanotransduction. Furthermore, this suggests that IFT88 and the cilium may be fundamentally important for purinergic-calcium signalling pathways.In several cell types fluid-flow deflection of primary cilia initiates a mechanotransduction pathway via polycystin 1 and 2 (PC1/2). In articular cartilage the chondrocyte primary cilium extends into cartilage matrix. Mechanical signals including compression and fluid flow trigger mechanosensitive regulation of matrix synthesis underpinning tissue homeostasis. Here we tested the hypothesis that the cilium plays a key role in chondrocyte mechanotransduction that includes ATP release, ATP-induced calcium transients and the subsequent regulation of matrix synthesis. These studies used murine chondrocytes with a hypomorphic mutation of"} +{"text": "MLL gene displayed extensive loss of methylation of an almost mutually exclusive set of CpGs, which instead affected introns and distal intergenic CpG islands and shores. When analyzed in conjunction with gene expression profiles, it became apparent that these specific patterns of DNA methylation result in differing roles in gene expression regulation. However, despite this subtype-specific DNA methylation patterning, a much smaller set of CpG sites are consistently affected in both AML subtypes. Most CpG sites in this common core of aberrantly methylated CpGs were hypermethylated in both AML subtypes. Therefore, aberrant DNA methylation patterns in AML do not occur in a stereotypical manner but rather are highly specific and associated with specific driving genetic lesions.We have developed an enhanced form of reduced representation bisulfite sequencing with extended genomic coverage, which resulted in greater capture of DNA methylation information of regions lying outside of traditional CpG islands. Applying this method to primary human bone marrow specimens from patients with Acute Myelogeneous Leukemia (AML), we demonstrated that genetically distinct AML subtypes display diametrically opposed DNA methylation patterns. As compared to normal controls, we observed widespread hypermethylation in IDH mutant AMLs, preferentially targeting promoter regions and CpG islands neighboring the transcription start sites of genes. In contrast, AMLs harboring translocations affecting the Acute myeloid leukemias (AML) are a group of malignancies that originate in the bone marrow. While many different genetic lesions have been linked to the different forms of this disease, it is also clear that these genetic lesions are not always sufficient to cause AML. DNA methylation plays a role in gene expression regulation, and abnormal distribution of DNA methylation has been observed in many cancers, including AML. Here we demonstrate that changes in DNA methylation in AML are not uniform across all AML subtypes, but rather they display unique patterns, which are closely linked to the underlying genetic lesions of each of the different forms of AML. Furthermore, these unique patterns of DNA methylation have different impacts on gene expression regulation in each AML subtype. HOXA9Acute myeloid leukemia (AML) is considered to be a genetically heterogeneous group of diseases, featuring functionally distinct somatic mutations and chromosomal translocations DNA methylation profiling of AMLs indicate that disruption of promoter cytosine methylation patterning is a universal feature of the disease. Promoter methylation signatures identify AML as composed of sixteen epigenetically defined subtypes We sought to examine quantitative, base-pair resolution DNA methylation patterns in clinical specimens with limited cell numbers, with adequate coverage of CpGs both within and outside of CpG islands. To accomplish this, we developed a modified version of the RRBS protocol, which retains its quantitative base-pair resolution while improving the coverage of regions outside CpG islands. We first validated the performance of the original RRBS assay using genomic DNA extracted from the HCT116 colon cancer cell line. We observed that RRBS yielded robust and reproducible results over a wide range of starting material ranging from 5 ng to 1000 ng without We next modified RRBS into a format that would be practical to perform in limited clinical specimens. First, we eliminated an intermediate clean-up procedure between the two rounds of bisulfite treatment in order to minimize sample loss during library preparation. Instead of two rounds of bisulfite conversion as previously described While RRBS has been shown to reliably detect gain of methylation, its ability to accurately detect genome-wide loss of methylation has not been extensively probed. Yet this is essential for clinical samples, since aberrant hypomethylation can be a dominant feature of tumor cells An increasing body of evidence demonstrates that biologically relevant changes in DNA methylation in cancer occur beyond CpG islands While the original RRBS alignment strategy used an MspI fragmented genome as a reference, whole-genome alignment strategies can also be applied to these data We previously reported that IDH-mut and MLLr AMLs distribute to different DNA methylation clusters and have distinct promoter DNA methylation signatures compared to normal CD34+ bone marrow controls (NBM) In order to identify the nature of the differences between IDH-mut and MLLr AMLs , the cytosine methylation profiles of these tumors were compared to normal CD34+ bone marrow cells from healthy donors (NBM), using logistic regression . In addition to statistical significance, we required a minimum cutoff of 25% methylation difference. This analysis revealed striking differences in the way that these two forms of AML differed from normal hematopoietic stem and progenitor cells. Specifically, we observed that IDH-mut AMLs display profound hypermethylation distributed across all chromosomes. In marked contrast, comparison of the cytosine methylation profiles of MLLr AMLs to NBM samples identified a predominance of aberrantly hypomethylated CpG site . More spEP300, CREBBP, FOS, JUN as well as several genes involved in regulation of apoptosis such as BAX, CASP3, CASP6 and TP73 and 26% in IDH-mut , and most of those DMCs were found at Alu elements (8% in IDH-mut and 10% in MLLr). Hypermethylated DMCs, on the other hand were depleted for repeat elements, with only 7 and 8% of hypermethylated DMCs overlapping with repeats in IDH-mut and MLLr, respectively . These were, mostly low complexity and simple repeats .Next we examined the common differentially methylated CpG sites in IDH-mut and MLLr AML . Of these, 76.6% (n\u200a=\u200a9148) were coordinately differentially methylated in the same direction in both AML subtypes, and the majority of these DMCs were aberrantly hypermethylated vs. NBMs . These concordantly hypermethylated DMCs were more frequently associated with polycomb repressive marks than concordantly hypomethylated DMCs . Concordantly hypermethylated CpGs were associated with genes involved in Cadherin, Wnt and Notch signaling pathways, many of which have been previously reported as frequently methylated in a variety of neoplasms, such as APC2 Different types of analyses and platforms used in previous studies have tended to favor either aberrant methylation of CpG islands We also observed significant differences in the absolute numbers of DMCs distributed to CpG islands and shores between the two subtypes . Specifically, we found that DMCs more frequently mapped to CpG islands in the IDH-mut AMLs cases (50% in IDH-mut vs. 29% in MLLr). In contrast, 50% of DMCs in the MLLr AMLs were found neither at CpG islands nor CpG shores but were instead annotated to regions even beyond CpG shores . These fWhen considering the relation of DMCs to RefSeq annotated genes we observed that approximately 40% of all DMCs in both AML subtypes were found within gene bodies. However, more detailed analysis identified markedly dissimilar regional distribution of DMCs between the IDH-mut and MLLr AMLs. First, MLLr AMLs displayed significantly more abundant DMCs at introns than IDH-mut AMLs (31 vs. 25%) and intergenic regions (39 vs. 35%). In contrast, promoter-associated DMCs were twice as frequent in IDH-mut AMLs (27 vs. 16%) Chi-squ. MoreoveWe then examined these regional differences in cytosine methylation relative to known regulatory elements. We compared the DMC sites from both IDH-mut and MLLr AMLs to available ENCODE data sets th percentile of expressed transcripts, Wilcoxon rank sum test p-value<0.005) . HoweverFurthermore, when examining DMCs and their correlation with differential gene expression between the different AMLs and the normal bone marrow specimens, we found that only DMCs at the core promoter regions were significantly inversely correlated with differential gene expression in IDH-mut AMLs . However, for MLLr AMLs, we observed that while core promoter DMCs were also significantly associated with differential expression , this association was also significant at upstream DMCs (up to 10 kb), both for DMCs located at CpG islands and CpG shores . Finally, downstream intronic DMCs overlapping with CpG islands also showed a significant correlation with expression in MLLr AMLs . Collectively, these findings suggest that subtype specific DNA methylation distribution in AMLs regulates gene expression in a subtype-defined manner. More precisely our data indicate a significant role for long-range epigenetic regulation in MLLr AML through distal intergenic and intronic CpG islands, whereas IDH-mut AMLs display a predominance of promoter-centric epigenetic regulatory effects.The study of gene promoters and CpG islands under the assumption that variation in the 5-methylcytosine status at these locations would have functional importance has been the focus of most cancer-related DNA methylation studies. Building upon the previously described RRBS method, the ERRBS assay described here made it possible to measure DNA methylation in primary AML samples beyond promoter regions, extending even into distal intergenic regions. This significantly enhanced genomic coverage allowed us to demonstrate that heterogeneity in epigenomic profiles in AML is not only a factor of different genes being affected, but rather encompasses a far more complex scenario, which includes the aberrant DNA methylation of distinct regions of the genome as well as differential mechanisms of regulation of gene expression according to genetic background. Given the specificity and reproducibility of these aberrant DNA methylation patterns, it is likely that their establishment in malignant cells is directly linked to genetic driver lesions. Our previous studies using HELP promoter microarrays are consistent with these results in that they revealed a hypermethylated promoter signature in IDH-mut AMLs, and a hypomethylated signature in MLLr. However those studies did not have the resolution or depth to reveal the true genomic nature, complexity and qualitative differences that we are now able to report regarding the nature of cytosine methylation distribution in these AML patients.Specifically, in the case of MLLr leukemias, aberrant DNA methylation consists mostly of aberrant hypomethylation of upstream and intronic CpGs including CpG islands and shores, but extending to and heavily involving even more distal regions. Hypomethylation of regulatory elements is consistent with the actions of MLL fusion proteins as transcriptional activators. However, in these tumors the distal localization of hypomethylation was more closely associated with the presence of HOXA9 and MEIS1 binding sites and enhancer regions than with MLL binding sites, suggesting that aberrant DNA hypomethylation in these tumors may be more closely related to effects of downstream targets of MLL than to the fusion protein itself. However, it should be noted that a subset of MLL peaks (6.5%) did indeed overlap with DMCs in the MLLr AMLs. Since our ChIP-seq antibody recognized both the wild-type and the rearranged copy of MLL. Given that MLL fusions have been shown to bind to a subset of wt-MLL target genes Even though the two AML subtypes were dramatically different, they still shared a core hypermethylated signature including genes previously shown to be almost universally hypermethylated in AMLs Most importantly, abnormal DNA methylation patterning does not occur in a stereotypical manner in cancer, but instead adopts distinct and specific distributions dependent at least in part on genetic background, even when comparing cases of the same tumor type with different driver mutations. Our analysis comparing gene expression and DNA methylation at base-pair resolution across three different sample types demonstrates that epigenetic regulation of gene expression in tumors may at least in part be context dependent, suggesting that cell-type specific factors may come into play to establish and maintain unique regulatory mechanisms in these cells. Finally, the large distances between DMCs and transcription start sites support a potential role for epigenome regulation at distal regulatory elements, via looping or other mechanisms, in directly influencing the specificity of the transcriptional machinery. Taken together our data support the existence of divergent roles of the epigenome in regulating the transcriptional profiles of AML and indicate that altered gene expression is associated with the differential methylation of distinct and non-overlapping CpGs and regions in tumors with different genetic backgrounds. Moreover, in the case of MLLr AMLs, these abnormal regulatory mechanisms extend far beyond the classically described cancer-associated promoter CpG island hypermethylation, and indicate that distal intergenic DNA methylation abnormalities may also have functional consequences in certain tumors. These findings are consistent with those described by other groups which have seen an association between differentially methylated regions at CpG shores in solid tumors and changes in gene expression 2. The HCT116 DNMT1(\u2212/\u2212) DNMT3b(\u2212/\u2212) double knockout clone number 2 (DKO) cell line was a kind gift from Dr. Steve Baylin. The cell line was grown in McCoys'5A medium with 10% FBS, 0.2 mg/ml Neomycin(G418), and 0.1 mg/ml Hygromycin B. Genomic DNA was extracted from the cell lines using standard phenol:chloroform extraction followed by ethanol precipitation.The human colorectal carcinoma cell line HCT116 was a kind gift from Dr. John Mariadason. The cell line was maintained in DMEM supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin and 100 \u00b5g/ml of streptomycin (Invitrogen) at 37\u00b0C and 5% COAML samples were obtained from previously reported, de-identified patient samples, from individuals enrolled in the Eastern Cooperative Oncology Group's (ECOG) E1900 clinical trial RRBS was performed as follows: i) 5, 50 or 1000 ng of high quality genomic DNA were digested with 200 U of MspI which cuts DNA regardless of cytosine methylation status at CCGG sequence in a 100 \u00b5l reaction for up to 16 hours at 37\u00b0C. DNA was isolated using standard phenol chloroform extraction and ethanol precipitation and resuspended into 30 \u00b5l of 10 mM TrIs pH 8.0.ii) End repair of digested DNA was performed in a 100 \u00b5l reaction using 15 U of T4 DNA polymerase (NEB M0203L), 5 U of Klenow DNA polymerase (NEB M0210L), 50 U of T4 Polynucleotide Kinase (NEB M0201L), 4 \u00b5l of premixed nucleotide triphosphates each at 10 mM (NEB N0447L) using T4 DNA ligase buffer with 10 mM dATP (NEB B0202S). The reaction was incubated at 20\u00b0C for 30 minutes and products were isolated using QIAquick PCR purification columns per manufacturer's recommended protocol (Qiagen) into 32 \u00b5l of EB buffer. iii) Adenylation was performed in a 50 \u00b5l reaction using 15 U Klenow fragment , 10 \u00b5l of dATP at 1 mM concentration using Klenow buffer (NEB). The reaction was incubated at 37\u00b0C for 30 minutes and products were isolated using MinElute PCR purification columns per manufacturer's recommended protocol (Qiagen) into 10 \u00b5l EB buffer. iv) Adenylated DNA fragments were ligated with pre-annealed 5-methylcytosine-containing Illumina adapters in a 20 or 50 \u00b5l reaction for 5 ng or 50 ng or higher starting materials respectively using 2000 U T4 DNA ligase (NEB M0202T) and 1.2 \u00b5M final concentration of methylated adapters at 16\u00b0C for a minimum of 16 hours. Products were isolated using MinElute columns per manufacturer's recommended protocol (Qiagen) into 10 \u00b5l EB buffer. v) Library fragments of 150\u2013175 and 175\u2013225 bp were gel isolated from a 1.5% agarose gel (using low range ultra agarose from Biorad) using the Qiaquick Gel Extraction kit per manufacturer's recommended protocol (Qiagen) into 40 \u00b5l EB buffer. vi) bisulfite treatment was performed using the EZ DNA Methylation Kit (Zymo Research) per manufacturer's recommended protocol with the following modifications: 1) incubation after the addition of CT conversion reagent was conducted in a thermocycler with the following conditions: 30 seconds at 95\u00b0C followed by 15 minutes at 50\u00b0C for 55 cycles and, 2) products were eluted into 40 \u00b5l nuclease free water. vii) PCR amplification for each library was performed in a 200 \u00b5l reaction containing 2 \u00b5l FastStart Hifidelity DNA Polymerase (Roche), 0.5 \u00b5M each of Illumina PCR primers PE1.0 and 2.0, 0.25 mM each nucleotide triphosphate using buffer 2 per manufacturer's recommendation and divided into four 50 \u00b5l reactions. The thermocycler conditions were: 5 minutes at 94\u00b0C, 18 cycles of 20 seconds at 94\u00b0C, 30 seconds at 65\u00b0C, 1 minutes at 72, followed by 3 minutes at 72\u00b0C. PCR products were isolated using AMPure XP beads per manufacturer's recommended protocol (Agencort) into 50 \u00b5l of EB buffer. viii) All amplified libraries underwent quality control steps including using Qubit 1.0 fluorometer and a Quant-iT dsDNA HS Assay Kit for quantitation (Invitrogen) and bioanalyzer visualization .Extended Reduced Representation Bisulfite Sequencing (ERRBS) was performed as described above for RRBS, except that in step number v, library fragment lengths of 150\u2013250 bp and 250\u2013400 bp were gel isolated.All data have been deposited for public access in the GEO database. The Accession number is GSE37454.The amplified libraries were sequenced on an Illumina Genome Analyzer II or HiSeq2000 per manufacturer's recommended protocol for 50 bp single end read runs. Image capture, analysis and base calling was performed using Illumina's CASAVA 1.7.Validation of select CpG methylation in HCT116 cell line was implemented by MALDI-TOF mass spectrometry using EpiTYPER by MassARRAY as previously described http://sourceforge.net/projects/theflexibleadap/). Adapter sequence contamination usually occurs towards 3\u2032ends of some reads. The adapter matching part of the read was removed if it aligned with the adapter sequence at least 6 base-pairs and had at most 0.2 mismatch error rate. Reads were aligned to whole genome using the bismark alignment software Reads were filtered from the adapter sequences using FAR software .http://www.r-project.org/) where we used 1-Pearson correlation distance and Ward's agglomeration method.Hierarchical clustering of the six samples was performed using the hclust function in R-2.14.0 between different samples were compared by taking the mean methylation percentage of CpG dinucleotides overlapping those regions. In order to calculate the correlation between different samples and generate the appropriate scatter plots we required that in any given region at least 3 CpG dinucleotides were covered by reads in both samples.The test for differential methylation was performed at the single base level. The test is performed only on CpG dinucleotides covered in all the test and control samples in each case. In order to improve the number of covered CpG dinucleotides across samples, we merged the read coverage on forward and reverse strand of a given CpG dinucleotide before doing the test. For the test, the number of methylated and unmethylated Cs aligning to each base were counted and compared across samples. To determine significant differential methylation between two sets of samples, we applied logistic regression and the likelihood ratio test. Observed p-values were adjusted with the SLIM method Pathway enrichment analysis was performed using the GREAT software Gene expression for IDH mutants and normal bone marrow cells are downloaded from the Gene Expression Omnibus (GEO) (accession: GSE24505). Normal bone marrow samples are not matched to the samples on this array however we averaged 5 normal bone marrow samples on the array to interpolate the expression profiles of our normal bone marrow samples. The sample matched gene expression profiles for cells with MLL translocation are downloaded from GEO (accession: GSE6891). Expression percentiles of each transcript are also calculated using R function \u201cecdf\u201d. The transcripts for each sample are divided into two categories high expressed (the top 15%) and low expressed (the bottom 15%).CpG islands are mapped to annotated transcripts for probes as follows. First, we mapped CpG islands to 10 kb window around the TSS of the annotated transcript, and CpG islands in this window are classified as TSS overlapping, upstream and downstream CpG islands depending on whether or not they overlap with TSS and relative location if they are not overlapping with TSS. Following that, we compared maximum methylation per island and maximum methylation per shore for high and low expressed genes on each sample. We used Wilcoxon's Rank sum test to compare maximum methylation distributions on each shore and CpG island for high and low expressed genes. For this comparison we only considered CpG islands and shores that have at least three genomic CpGs covered by bisulfite reads.When correlating DMCs with the differential expression, we first calculated fold-change of MLLr vs. NBM and IDH-mut vs. NBM samples. Expression data for NBM samples were available for both IDH-mut and MLLr fold-change calculations within the respective microarray types and downloaded from GEO (accession numbers GSE24505 and GSE6891 respectively). We calculated fold-change between the average expression values of the groups. Following that we measured correlation between percent methylation difference at DMCs and fold-change of the nearest gene (obtained by extracting the nearest TSS) using \u201ccorrelation.test\u201d in R. We performed separate correlation analyses for DMCs at the core promoter , upstream from the TSS (up to 10 kb), within CpG islands (up to 5 kb from TSS), within CpG island shores (up to 5 kb from TSS), within intronic regions, at intronic CpGs, and at CpGs within intronic CpG islands and shores.MLL ChIP-seq experiments were performed in the MLL-AF4 cell line RS4;11 (ATCC#CRL-1873) using antibodies to MLL1 (Bethyl Laboratories A300-086A). ChIP-seq libraries were prepared from 10 ng of immunoprecipitated material using Illumina's ChIP-seq kit as per manufacturer's instructions, and then sequenced on a Genome Analyzer IIx sequencer. Alignment against the human genome, peak calling and downstream analysis was performed using ChIP-seeqer The ENCODE CTCF, H3K27me3, H3K4me1 and H3K4me3 peak locations are downloaded using UCSC table browser Figure S1ERRBS is highly reproducible and sensitive. (A) Correlation between CpG dinucleotides, CpG islands and promoter methylation levels using pearson correlation between technical replicas of ERRBS using 5, 50 or 1000 ng genomic DNA from the HCT116 cell line. (B) Distribution histograms of CpG coverage and CpG methylation levels along forward and reverse strands in HCT116 ERRBS results. (C) Distribution histogram of CpG methylation levels along forward and reverse strands in DKO ERRBS results. Similar results were obtained from reverse strand (data not shown) and CpG coverage distributions over both strands were similar to coverage seen with HCT116 sequencing (data not shown). (D) Technical validation of ERRBS performance in HCT116 at select CpGs by MassARRAY. Dot plot shows correlation between DNA methylation as measured by ERRBS (x-axis) and percent methylation as measured by MassARRAY EpiTyper (y-axis). (Correlation coefficient: 0.97).(TIF)Click here for additional data file.Figure S2Biological replica reproducibility. (A) Correlation plot of CpG dinucleotide methylation levels between two biological replica of ERRBS data using normal bone marrow controls (NBM_#1 and NBM_#2_Rep#2). (B) Correlation plot of CpG dinucleotide methylation levels between two biological replica of ERRBS data using IDH mutant AML samples (IDH-mut_#1 and IDH-mut_#2). (C) Correlation plot of CpG dinucleotide methylation levels between two biological replicas of ERRBS data using MLL translocated AML samples (MLLr_#1_Rep#2 and MLLr_#2).(TIF)Click here for additional data file.Figure S3DNA methylation patterns naturally segregate AML and NBM samples. Unsupervised analysis using either principal component analysis or hierarchical clustering of DNA methylation by ERRBS at (A) all CpGs, (B) non-promoter CpGs, (C) non-promoter intron CpGs, (D) CpGs within CpG islands and (E) CpGs within CpG shores, segregates the samples into their three biological groups.(TIF)Click here for additional data file.Figure S4Differential methylation in MLLr and IDH-mut AMLs are preserved at 40% and 10% cutoffs. Chromosome ideogram representing differential methylation in IDH-mut AMLs vs. NBM (A) and MLLr AMLs vs. NBM (B), using changes greater than 10%. Light and dark magenta points represent hypermethylation changes relative to NBM of 10\u201340% and greater than 40% respectively. Light and dark green points represent hypomethylation changes relative to NBM of 10\u201340% and greater than 40% respectively.(TIF)Click here for additional data file.Figure S5Percentage of DMCs overlapping with repeats. Bar plots showing percentage of hyper- (magenta) and hypo-methylated (green) DMCs on repeat regions. Overall, 24\u201326% of hypo-methylated DMCs and \u223c7% of hyper-methylated DMCs overlap with repeats. 10.7% of hypo-methylated DMCs of MLLr overlap with Alu repeats and 8.6% of hypo-methylated DMCs of IDH-mut overlap with Alu repeats.(TIF)Click here for additional data file.Figure S6DNA methylation and gene expression relationships display subtype-specific differences. CpG islands and shores across the genome were categorized into those located upstream from a transcription start site (TSS), overlapping a TSS or located downstream from a TSS. Boxplots are plotted that illustrate the maximum DNA methylation levels at these CpG islands and CpG shores for the high expressed genes (top 15th percentile expressed genes) and the low expressed genes (the bottom 15th percentile expressed genes). Each row is for a different sample: Normal bone marrow (top); IDH-mut AML (middle) and MLLr AML (bottom). The boxplots are color-coded depending on the expression status of associated genes. Significantly different distributions are marked with a star.(TIF)Click here for additional data file.Table S1Summary of RRBS and ERRBS experiments. All sequencing was performed using either the Illumina Genome analyzer II or HiSeq2000 . We routinely acquired >40 million reads per sample, with alignment rates ranging from 55\u201370%. Shown are the number of CpGs covered, bisulfite conversion efficiency and mean CpG coverage rates for each sample.(DOCX)Click here for additional data file.Table S2Methylation sequencing by MassARRAY EpiTYPER. MassARRAY was performed on bisulfite-converted DNA from HCT116 using the following primers targeting the listed amplicons.(XLSX)Click here for additional data file.Table S3Pathway analysis of DMCs in AML subtypes. Pathway enrichment analysis was performed using GREAT. Enriched terms in PANTHER Pathways are shown with their hyper-geometric test and binomial test q-values. (A) Pathway analysis for uniquely hyper-methylated DMCs in IDH-mut AML samples. (B) Pathway analysis for uniquely hypo-methylated DMCs in MLL-r AML samples. (See separate excel spreadsheet for full list of genes in each pathway).(XLSX)Click here for additional data file.Table S4Pathway analysis of concordantly hypermethylated DMCs in AML subtypes. Pathway enrichment analysis was performed using GREAT. Enriched terms in PANTHER Pathways are shown with their hyper-geometric test and binomial test q-values. Results from pathway analysis for concordantly hypermethylated DMCs in IDH-mut and MLL-r AML samples are listed.(DOCX)Click here for additional data file.Table S5Genes with recurrent aberrant DNA methylation by HELP that were validated by ERRBS. Listed are the fifteen genes covered by both assays that were hypermethylated in the current study.(DOCX)Click here for additional data file."} +{"text": "Purpose: Higher order chromatin structure progressively changes with cell differentiation and seems to play an important role in DNA double-strand break (DSB) induction and repair (reviewed in [1]). We compared DNA damage in heterochromatin (Hc) upon the action of qualitatively different radiations. We also studied, how is the sensitivity to DSB induction, assembly of repair foci and processing of DSBs influenced by the differentiation-induced changes in chromatin structure and composition.Materials and methods: Formation, localization (relative to higher-order chromatin domains) and mutual colocalization of \u03b3H2AX and p53BP1 repair foci have been studied together with DSB repair kinetics in spatially fixed human skin fibroblast and differently differentiated white blood cells (WBC) irradiated with gamma rays, protons of different energies , and 20Ne ions (submitted). Immunostaining and ImmunoFISH were used in combination with high-resolution confocal microscopy and living cell imaging [4].Results: We found that less DSBs appear in Hc after irradiating cells with gamma rays and protons but not 20Ne ions (preliminary results). In addition, contrary to \u03b3-irradiated human skin fibroblasts and lymphocytes, mature granulocytes neither express DSB repair proteins nor form functional repair foci [5]. At least some DSB repair proteins (e.g. 53BP1) are expressed and \u03b3H2AX foci still occur in immature granulocytes and monocytes ; however, the colocalization of \u03b3H2AX with 53BP1 is low and the majority of DSBs are not repaired. Despite this fact, \u03b3H2AX foci protrude from Hc into nuclear subcompartments with low chromatin density. Our living cell observations suggest that 53BP1 can penetrate into the interior of dense Hc domains only after their decondensation [2].Conclusions: We show that Hc is less sensitive to DSB induction by gamma rays but not heavy ions; lower Hc hydratation and higher protein density (when compared with euchromatin) probably reduce formation of free radicals and increase their sequestration, respectively. This mechanism can protect cells against the indirect effect of ionizing radiation (marked for gamma rays and protons but not heavy ions). Hc features, however, preclude DSB repair, which is best illustrated by its absence in differentiated WBC but not their immature precursors. The protrusion of Hc-DSBs into low-density chromatin nuclear subdomains, however, appears also in differentiated WBC, so the process might simply follow physical forces (e.g. as suggested by M Durante's group).There is no Clinical Trial Registration number."} +{"text": "To compare single photon emission computed tomography (SPECT) and cardiac magnetic resonance (CMR) myocardial perfusion imaging for the detection of myocardial ischaemia in patients with triple vessel coronary artery disease (3VD) from a prospectively acquired cohort of patients with suspected coronary heart disease (CE-MARC study).3VD is associated with an adverse prognosis, which may be ameliorated by coronary artery revascularisation. Some patients with 3VD may not be identified by visual myocardial perfusion analysis due to the phenomenon of balanced ischaemia. CMR first-pass perfusion with its superior spatial resolution may be more effective than SPECT in identifying 3VD.Thirty-nine patients with 3VD at X-ray coronary angiography and 39 matched patients with no significant coronary disease were identified from the CE-MARC study population . PatientBy per-patient analysis the sensitivity and specificity of visual CMR and SPECT analyses for the detection of ischaemia did not differ significantly (92% and 97% vs 82% and 91% respectively). Among the 3VD patients, 34% had hypoperfusion in all three coronary artery territories by visual CMR analysis and 18% by SPECT (p=0.14). There were significant differences in mean transmural MPR and subendocardial MPR between 3VD and normal patients respectively. Receiver operator characteristic curve analysis produced an area under curve of 0.94 for the mean MPR of all three slices and 0.92 for midventricular slice subendocardial values alone (p=NS). A mid-slice subendocardial MPR cut-off of 1.76 yielded 97% sensitivity, 70% specificity and 96% negative predictive value for the detection of ischaemia. Figure Patients with 3VD may be reliably differentiated from those without obstructive coronary disease by visual or quantitative CMR myocardial perfusion assessment. Visual CMR analysis performs at least as well as SPECT imaging in this regard. Fermi-derived MPR assessment may be performed efficiently using the middle slice alone without a loss of diagnostic accuracy relative to a three-slice analysis."} +{"text": "MYBPC3 encoding cardiac myosin binding protein C are common genetic cause of hereditary cardiac myopathies. An intronic 25-bp deletion in MYBPC3 at 3\u2032 region is associated with dilated (DCM) and hypertrophic (HCM) cardiomyopathies in Southeast Asia. However, the frequency of MYBPC3 25 bp deletion and associated clinical presentation has not been established in an unrelated cohort of left ventricular dysfunction (LVD) secondary to coronary artery disease (CAD) patients.Mutations in MYBPC3 25 bp polymorphism on LVD in two cohorts of CAD patients.We sought to determine the role of MYBPC3 25 bp polymorphism was determined by polymerase chain reaction. Our results showed that carrier status of MYBPC3 25 bp deletion was associated with significant compromised left ventricle ejection fraction (LVEF \u226445) in CAD patients . To validate our results, we performed a replication study in additional 140 cases with similar clinical characteristics and results again confirmed consistent findings . Also, presence of the gene deletion did not have significant association in CAD patients with preserved ejection fraction (LVEF>45) .The study included 265 consecutive patients with angiographically confirmed CAD and 220 controls. MYBPC3 DW genotype and D allele was associated with compromised LVEF implying that genetic variants of MYBPC3 encoding mutant structural sarcomere protein could increase susceptibility to left ventricular dysfunction. Therefore, 25 bp deletion in MYBPC3 may represent a genetic marker for cardiac failure in CAD patients from Southeast Asia.The frequency of Coronary artery disease (CAD) is defined as atherosclerotic blockageTNNI3),MYBPC3)Earlier genetic studies have correlated LVD with dilated cardiomyopathy (DCM).MYBPC3 result in disorganized sarcomeric structure, thus MYBPC3 has emerged as an important gene for increased risk of heart failure through cardiomyopathies (DCM and HCM).Myosin-binding protein C (MyBP-C) is a key constituent of cardiac muscle thick filaments, which by interacting with myosin,MYBPC3 at the 3\u2032 region of the gene MYBPC3 has any role in LV dysfunction. To the best of our knowledge, it is the first study to report the association of 25 bp polymorphism of MYBPC3 with low left ventricular ejection fraction in CAD patients.A common polymorphic intronic deletion of 25-base- pair in Demographic and lipid profile of healthy subjects is shown in MYBPC3 gene deletion in controls were in accordance with Hardy-Weinberg equilibrium (p\u200a=\u200a0.98). Considering the low allelic frequency allele, we also applied fisher exact test for conditional probability of heterozygous variants which was found to be p\u200a=\u200a0.97.Distributions of genotypes of MYBPC3 25 bp polymorphism between CAD patients (both stages) and healthy controls. Our results suggested significant association of MYBPC3 gene deletion with CAD ; D allele p value \u200a=\u200a <0.001 [9.8 (3.5\u201327.6)] while no risk of DW genotype was observed in CAD patients with preserved ejection fraction : 403 bp and mutant [DW]: 378 bp) were analysed on 6% polyacrylamide gel. Ten percent of samples from patients and controls were sequenced to evaluate the quality of genotyping, which showed 100% concordance.Genomic DNA was isolated from peripheral blood leukocytes according to a standard salting out method. MYBPC3 25 bp deletion and significant risk factors of CAD were analyzed using binary logistic regression. Student t test was applied to find the significance for plasma cholesterol. Differences between groups stratified on the basis of genotypes were assessed using z proportional test. Overall we performed meta analysis by Stouffers method to combine the results of two cohorts (primary and replication) to calculate the overall z and p values. A two-tailed p-value of less than 0.05 was considered a statistical significant result. All statistical analyses were performed using SPSS software version 16.0 .Descriptive statistics were presented as mean and standard deviation (SD) for continuous measures while absolute value and percentages were used for categorical measures. The chi-square goodness of fit test was used for any deviation from Hardy Weinberg Equilibrium in controls and fisher exact test was applied as given by Emigh"} +{"text": "Revascularisation of patients with low ejection fraction has beneficial effects. It prevents progressive left ventricular dilatation, sudden cardiac death, reduction of functional capacity.Thirty-seven consecutive patients with low EF (\u226430%) who underwent CABG in our department between February 2010 and April 2012 were analyzed retrospectively . Short and mid term results of CABG on functional capacity and angina were evaluated. Functional capacity and angina were ranked according to the New York Heart Association (NYHA) and Canadian Cardiovascular Society (CCS) classification systems consecutively. Eight patients underwent of-pump.In-hospital mortality was 5.4%. One patient died seven months after hospital discharge and mortality was due to heart failure. Annual mortality was 8.1%. The patients did have significantly decreased angina and increased functional capacity after the operation. Sixteen patients were in CCS class I/II and 19 patients were in CCS III/IV preoperatively while all patients were in CCS class I/II postoperatively. Eighteen patients were in NYHA class I/II and 17 patients were in NYHA class III/IV preoperatively while all patients were in NYHA class I/II postoperatively. Postoperative mean values of CCS angina and NYHA functional capacity classifications were significantly lower than preoperative scores . Postoperative mean value of CCS angina class improvement found as 1,35\u00b10,37 and mean value of NHYA functional capacity improvement was 1,33\u00b10,25 when compared to preoperative status. Six and 12-month outpatient CCS angina class and NHYA functional capacity did not show any significant difference.Short and mid term follow-up of patients with low EF who underwent CABG, revealed that CCS angina and NYHA functional capacity were improved significantly when compared to their preoperative status."} +{"text": "The geometry about the Zn atom is distorted tetra\u00adhedral, with the largest deviation observed in the magnitude of the Cl\u2014Zn\u2014Cl angle. Similar distortions are observed in the cobalt analogue and related zinc compounds containing metallocyclic rings with more than six members. The copper analogue exhibits a more severe distortion of the metal coordination sphere than is observed in the title compound.The title zinc complex, [ZnCl DOI: 10.1107/S1600536811046368/lr2035Isup2.hkl Structure factors: contains datablock(s) I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "Phlebotomus perfiliewi sandflies collected in Serbia in 1976. YBV shows very low nucleotide identities to other members of the Vesicular stomatitis virus serogroup and does not contain a reading frame for C\u2032/C proteins.We present pyrosequencing data and phylogenetic analysis for the full genome of Yug Bogdanovac virus (YBV), a member of the Vesicular stomatitis virus serogroup of the Rhabdoviridae isolated from a pool of Phlebotomus perfiliewi sandflies collected in Serbia. Electron microscopy and serological analysis placed it into the Vesicular stomatitis virus (VSV) serogroup of the Rhabdoviridae. Antibodies against YBV were found in humans (6/274 tested) [Vesiculovirus of rhabdoviruses was isolated in 1976 from a pool of 200 unengorged female tested) but the mans 6/27 tested [ tested) , 9, two tested) \u201312 was d tested) . In orde tested) . A MID-b tested) ), VSIV ("} +{"text": "Chronic cerebral hypoperfusion is believed to be a critical factor on the occurrence of AD and VaD for which there is no effective therapy currently. It has been reported that high levels of circulating high density lipoprotein (HDL) can reduce the risk of cardiovascular disease. Although the relationship between cognitive function and serum lipids has captured attention, previous studies have shown ambiguous results. We investigated the effects of Curcumin on cognitive impairment and serum HDL levels and investigated whether increasing serum HDL levels ameliorate VAD-like cognitive deficits in rats induced by permanent occlusion of bilateral common carotid arteries (2VO).Male Sprague-Dawley rats were subjected to permanent occlusion of bilateral common carotid arteries (2VO) to produce chronic cerebral ischemia. Animals were randomly divided into 5 groups: normal control group, sham-operated group, 2VO group, 2VO+Curcumin100mg/kg group, 2VO+ Curcumin50mg/kg group. Low doses of Curcumin (50mg/kg) or high doses of Curcumin (100mg/kg) were dissolved in DMSO. All animals were injected intraperitoneally with DMSO solution of Curcumin or a same volume of normal DMSO after surgery. Each group was injected once daily for four consecutive weeks. The spatial learning capacity and cognitive function of these animals was assessed in the Morris water maze 30 days after the onset of 2VO. Serum concentrations of high-density lipoprotein (HDL) were measured. Associations between serum concentrations of HDL and cognitive function were investigated for each group.Morris water maze for spatial orientation abilities showed that rats in 2VO group develop impaired learning and cognitive deficits, injection intraperitoneally of Curcumin can attenuate cognitive impairment ( P<0.05). 2VO+Curcumin group had higher serum HDL cholesterol levels than that of 2VO group (P<0.05). No significant differences were found in the serum level of HDL among 2VO+Curcumin group rats, sham-operated group rats and normal control group rats. Further more, plasma HDL levels correlated significantly with cognitive function.Our findings suggest that Curcumin have a protective influence on cognitive function and apparently increase serum HDL in 2VO group rats. Furthermore, serum HDL levels have positive association with cognitive function. The relation between Curcumin and HDL is hoped for new preventive and therapeutic strategies for cognitive deficits."} +{"text": "Duchene Muscular Dystrophy (DMD) is a lethal disease caused by ubiquitous lack of dystrophin, but the interaction with regional cardiac mechanical forces that may facilitate eventual expression of abnormal contractile function is unknown. Diffusion tensor MRI (DTI) was used to evaluate function in Langendorff perfused hearts in the mdx mouse model of DMD. Abnormal calcium kinetics and sheet mechanics (by DTMRI) occurred more prominently at the mid-upper ventricle, suggesting that regional mechanics influence the development of heart failure.The relationship between abnormal sarcoplasmic Ca2+ handling and cardiac structural mechanics in mdx mice has not been defined. DTI has been used to quantify the contribution of myocardial sheet architecture in ventricular wall thickening mechanics. In this study, DTI was employed to assess altered sheet mechanics that might be influenced by impaired calcium homeostasis. Furthermore, optical mapping of AP and CaT were employed to characterize local difference in ex vivo beating hearts.Sixteen-month old mdx (n = 10) and age matched wildtype mouse hearts were prepared for Langendorff perfusion by sequentially cardiac arrest in diastole and systole for DTI: The first group (WT-NC & mdx-NC) was perfused with regular St. Thomas\u2019 cardioplegic solution containing normal [Ca2+] (1.2 mM) to arrest hearts in diastole. To assess the regulatory effect of [Ca2+] on diastolic sheet mechanics, the second group (WT-LC & mdx-LC) was perfused with modified cardioplegic solution containing low [Ca2+] (0.078 mM). All hearts were then reperfused with Krebs buffer to resume beating followed by 2.5mM barium-induced systolic arrest for DTI. Absolute values of sheet angles, |\u03b2|s, in each heart were calculated from the diffusion tensors. MANOVA was used for statistical analysis. Another set of Langendorff beating mouse hearts from WT (n=2) and mdx (n=3) mice were optically mapped. RH237 and Rhod-2 AM were used as membrane potential and calcium probes. Steady state restitution pacing protocol was applied to estimate the calcium dynamics at different heart rates.In diastole, mdx hearts exhibited lower |\u03b2| in mid-upper ventricle than WT did. Reducing Ca2+] in cardioplegic solution normalized the diastolic |\u03b2| in mdx hearts but had no detectable effect on WT hearts. No significant difference of systolic |\u03b2| was observed between mdx and WT hearts. Fig. Optical + in cardThe observed lower diastolic |\u03b2| in mdx hearts indicates that mdx cardiomyocytes fail to fully relax in diastole. The regulatory effect of [Ca2+] on diastolic |\u03b2| of mdx hearts suggests that the myocardial sheet diastolic dysfunction is reversible despite the advance age of mdx mice. Optical mapping of CaT in beating hearts further confirmed the DTI-detected localized calcium-dependent abnormality, indicating that the ubiquitous dystrophin deficiency in mdx mice depends on regional cardiac features for its affection. In conclusion, the disturbed sheet mechanics may reflect functional consequences of abnormal Ca2+ handling in vivo, which likely correlates with membrane calcium channel dysfunction that precedes eventual cardiac failure.RO1 HL073646."} +{"text": "Risk for Ovarian Malignancy Algorithm (ROMA) and Human epididymis protein 4 (HE4) appear to be promising predictors for epithelial ovarian cancer (EOC), however, conflicting results exist in the diagnostic performance comparison among ROMA, HE4 and CA125.Remote databases and full texts bibliography were searched for relevant abstracts. All studies included were closely assessed with the QUADAS-2 . EOC predictive value of ROMA was systematically evaluated, and comparison among the predictive performances of ROMA, HE4 and CA125 were conducted within the same population. Sensitivity, specificity, DOR (diagnostic odds ratio), LR\u2009\u00b1\u2009(positive and negative likelihood ratio) and AUC (area under receiver operating characteristic-curve) were summarized with a bivariate model. Subgroup analysis and sensitivity analysis were used to explore the heterogeneity.Data of 7792 tests were retrieved from 11 studies. The overall estimates of ROMA for EOC predicting were: sensitivity , specificity , and AUC . Comparison of EOC predictive value between HE4 and CA125 found, specificity: HE4 \u2009>\u2009CA125 ; AUC: CA125 \u2009>\u2009HE4 . Comparison of OC predictive value between HE4 and CA125 found, AUC: CA125 \u2009>\u2009HE4 . Comparison among the three tests for EOC prediction found, sensitivity: ROMA \u2009>\u2009HE4 ; specificity: HE4 \u2009>\u2009ROMA \u2009>\u2009CA125 .ROMA is helpful for distinguishing epithelial ovarian cancer from benign pelvic mass. HE4 is not better than CA125 either for EOC or OC prediction. ROMA is promising predictors of epithelial ovarian cancer to replace CA125, but its utilization requires further exploration. Ovarian cancer is the leading cause of death from gynecologic cancers in the United States and the fifth-top cause of cancer death in women Link 1). Non-specific clinical manifestation mainly hinders the early diagnosis of ovarian cancer. Non-spe, it incrWith high-throughput technologies employed, a large number of new biomarkers have been discovered -10. HumaMoore and colleagues have expth December, 2011) were searched. Reference lists of articles identified were manually searched. Publication languages were not limited. The terminology for search was based on the standardized National Library of Medicine MeSH terms and free texts. The search strategies of all the databases were based on those of PubMed and the Two authors (RXT and WPL) independently screened the search results based on the titles and abstracts. The full text of selected articles were reviewed independently by another two authors (KC and LLY) to determine the inclusion. Disagreements were resolved by referring to a third author (MC).Studies that investigated both serum HE4 and CA125 as diagnostic tests or calculated the ROMA algorithm were included if (1) they were cross-sectional studies; and (2) performed in the same population presenting pelvic mass; (3) all serum specimens were collected preoperatively; (4) all subjects with histological diagnostic information; (5) with sufficient data for reconstructing fourfold table.Studies recruiting participants without presenting pelvis mass, with obviously error data or ROC curve analysis containing healthy person and case\u2013control studies were excluded. Case\u2013control studies were excluded, for these studies had a tendency of overestimating or underestimating the diagnostic performance of a test .The data extracted from each study included: author; year; country; design; recruitment; age; menopausal status; test methods (e.g. chemilumenesence immunoassay); number of patients; sensitivity; specificity and cut-off value. Four fold tables were reconstructed. Two reviewers (FKL and RXT) independently extracted the data for each study and referred to a third opinion (MC) when disagreements appeared. Important data that were not provided in the original studies were referred to their authors through Emails.Since the Risk of Ovarian Malignancy Algorithm (ROMA\u2122) is a qualitative serum test that combines the results of HE4 EIA (enzyme immunometric assays), ARCHITECT CA 125 II\u2122 and menopausal status into a numerical score. Index tests for HE4 and CA125 in this meta-analysis questions were specified as EIAs and chemilumenesence immunoassays respectively. ROMA algorithm is the following :Reference standard was based on outcomes of histopathological diagnosis. In all studies, ovarian cancer surgical stages were referred to criteria from FIGO (Link 3)The methodological quality of each study was evaluated with QUADAS-2 quality Index tests for HE4 and CA125 in this meta-analysis questions were specified as EIAs and chemilumenesence immunoassays respectively. For tests of HE4, the chemilumenesence immunoassays were more sensitive than the specified EIAs, thus bias might be introduced into pooling of studies. And similarly, for CA125, EIA and RIA (radioimmunoassay) assays were less sensitive and steady than chemilumenesence immunoassays, so studies using either EIA or RIA will be considered as High Concern Regarding Applicability. The ROMA test employed the results from tests of CA125 and HE4 within the same study. So ROMA was considered as High Concern Regarding Applicability when either HE4 or CA125 test was evaluated as High Concern Regarding Applicability.The statistical analysis is based on the following steps: (1) qualitatively describing the findings; (2) searching for heterogeneity and threshold effect; (3) figuring out the sources of heterogeneity by subgroup analysis; (4) choosing appropriate model and pooling estimates statistically. Univariate and biva2 estimates 1 What are the key statistics about ovarian cancer? American Cancer Society Web site. [. Handbook for DTA Reviews. Cochrane Collaboration Web site. [http://srdta.cochrane.org/handbook-dta-reviews].2 Diagnostic Test Accuracy Working Grouphttp://www.figo.org/]3 International Federation of Gynecology and Obstetrics FIGO website [The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2407/12/258/prepubTable S1. Searching strategies.Click here for fileFigure S1. Deeks\u2019 funnel plot for ROMA. Click here for fileTable S2. Influence analysis of individual studies for diagnostic performance of ROMA. Estimates were pooled by bivariate model. Excluding any individual study only a small change were resulted in the sensitivity (sen), specificity (spe) or diagnostic odds ratio (DOR) compared with all eligible studies. All differences were not significant (p\u2009>\u20090.05).Click here for fileFigure S2. Forest Plots for comparison between HE4 and CA125 for premenopausal and postmenopausal women and early stage EOC groups.Click here for file"} +{"text": "The prevalence of diabetes (DM) and its associated manifestations is higher in New Zealand M\u0101ori than New Zealand Europeans. There is no current evidence regarding podiatric clinical characteristics of Maori with diabetes. This presentation highlights findings from a group of Maori with diabetes that have foot pathology as determined by a national generic general practice review.Fifty three patients with diabetes were recruited from two M\u0101ori Primary Health Organisations. Podiatric specific characteristics, patient demographics and general medical conditions were recorded.Fifty three (n=53) M\u0101ori patients were recruited for this study with a median age of 54 years (IQR: 45.5 \u2013 61.5 years). We found that M\u0101ori had a median duration of diabetes for 15 years, were obese (median body mass index 35.7) with sub-optimal glycaemic control (HbA1c >8% 42%). Podiatric specific characteristics revealed good arterial flow and a median neuropathy score of 2 . Half the cohort displayed restriction in movement of the right ankle joint (n= 31) and first metatarsophalangeal joints (n=29). Foot education was favourable with positive benefits such as daily foot inspection (n=52). Using a modified classification tool which targets the risk factors collectively, thirty two (60%) patients were identified as requiring regular podiatry management.A standardised evidence-based screening and assessment tool could be widely applied by primary care health podiatrists in the detection of imminent diabetes-related foot pathology and to support disease management. This collective assessment would be of particular benefit to communities, including Maori; where the incidence of diabetes and its complications is higher."} +{"text": "The chemical mechanisms associated with Ir(piq)2(tmd) are investigated by ESR spin-trapping, laser flash photolysis, steady state photolysis, cyclic voltammetry and luminescence experiments.In the present paper, the photoredox catalysis is presented as a unique approach in the field of photoinitiators of polymerization. The principal photocatalysts already reported as well as the typical oxidation and reduction agents used in both reductive or oxidative cycles are gathered. The chemical mechanisms associated with various systems are also given. As compared to classical iridium-based photocatalysts which are mainly active upon blue light irradiation, a new photocatalyst Ir(piq) Photoredox catalysis is now well-known and largely used in organic synthesis, especially in the development of sustainable radical-mediated chemical processes under very soft irradiation conditions , e.g., enantioselective alkylation, cycloaddition, etc. \u201314. RuthPhotoredox catalysis was then introduced into the polymer photochemistry field (area) in the very past years (see a review in \u201322). IndPIs and PISs have been extensively developed both in industrial R&D and academic laboratories \u201340. PIs This approach opened up \u201353 a newAlmost no photoinitiator is consumed.Since the spectral photosensitivity extends now from the UV to the visible, laser excitation in the purple, blue, yellow, green or red is feasible.Low light intensities can be used; this is a catalytic process without loss of efficiency with irradiation.Photopolymerization under sunlight becomes reachable.The production of the radical or cationic initiating species for the FRP of acrylates or the FRPCP of epoxides, respectively, is quite easy; polymerization of sustainable monomers can also be achieved .A possible dual behavior is achieved.N-vinylcarbazole) or PIC/amine/alkyl halide. Also, relatively high intensity light sources can be obviously employed. However, and with more interest, very soft irradiation conditions are also suitable to polymerize radical and cationic monomers (see Examples of PICs proposed by us in the photopolymerization reactions are depicted in mers see below unThe present paper will i) review the various possible mechanistic schemes using metal and metal free oxidizable or reducible PICs for FRP, CP and FRPCP, ii) show examples of reported PICs, iii) discuss the encountered mechanisms in various PIC based photoinitiating systems, iv) highlight some examples of high performance PISs and v) present the efficiency of a novel PIC in photopolymerization reactions as well as the excited state processes involved in the photoinitiating systems used.i) Compounds: The synthesis of bisiridium is described below (following a published procedure). All reagents and solvents were purchased from Aldrich or Alfa Aesar and used as received without further purification.ii) Irradiation sources: Several lights were used: 1) polychromatic light from a halogen lamp ; 2) monochromatic light delivered by a laser diode at 532 nm (I0 \u2248 100 mW/cm2) and 3) LEDs at 514 nm.iii) Free radical photopolymerization (FRP) of acrylates: The experiments were carried out in laminated conditions or under air. The prepared formulations deposited on a BaF2 pellet (25 \u00b5m thick) were irradiated (see the irradiation devices). The evolution of the double bond content was continuously followed by real time FTIR spectroscopy (JASCO FTIR 4100) at about 1630 cm\u22121 as in ..56].N-vinylcarbazole NVK (3SiH) and exhibits a relatively similar performance in photoinitiating systems of cationic polymerization )3+) and a phenacyl radical is produced upon the subsequent cleavage of the phenacyl halide radical anion. Later on, other amines and bromides were proposed for the photocatalytic radical polymerization of various methacrylates ..66].All these reactions allowed the radical photopolymerization of formulations containing multifunctional synthetic acrylates using UV to red lights delivered by low intensity sources.3 ) is characterized by an intense absorption at 370 nm and exhibits an only moderate absorption at \u03bb > 400 nm. Among others, the design of green light sensitive Ir complexes is an interesting challenge. This is the reason why the new Ir based PIC iridium noted Ir(piq)2(tmd); There is still a need for the development of new PICs with i) improved light absorption properties and ii) high reactivity. These new PICs can be highly worthwhile for the use of very soft irradiation conditions. For example, the actual Ir and Ru-based PICs are mainly characterized by a blue light absorption and are operative in the 400\u2013480 nm spectral range, i.e., the well-known and commercially available Ir derivative Ir(ppy)2(tmd) are much better than those of Ir(ppy)3 (2(tmd) is perfectly adapted for applications under green light exposure (LED bulb at 514 nm or a laser diode at 532 nm) and exhibits an almost panchromatic behavior in the 400\u2013650 nm range.Indeed, the visible light absorption properties of Ir(piq)Ir(ppy)3 . Remarka2(tmd) by laser flash photolysis experiments. A relatively similar lifetime (1.3 \u03bcs) was previously measured for 3Ir(ppy)3 [A triplet-state lifetime of 1.1 \u03bcs has been determined for Ir(piq)Ir(ppy)3 . Such lo2(tmd) is characterized by an oxidation potential of 0.67 V vs SCE (ET ~ 2.07 eV) was determined from absorption/luminescence experiments (3Ir(piq)2(tmd)/Ph2I+ oxidation process can be expected, i.e., the free energy change \u0394G for the corresponding electron transfer is negative 2(tmd), the reduction potential of Ph2I+, the excited triplet state energy of Ir(piq)2(tmd), and the electrostatic interaction energy for the initially formed ion pair, generally considered as negligible in polar solvents) [Ir(piq)V vs SCE ; its trieriments . From tholvents) .3PIC/Ph2I+ interaction is also in line with a fast photolysis of the PIC observed during the irradiation of Ir(piq)2(tmd)/Ph2I+ (2(tmd)/Ph2I+ in the presence of phenyl-N-tert-butylnitrone (PBN), phenyl radicals are also detected can be used in an oxidative cycle in combination with Ph2I+ and NVK.This favorable d)/Ph2I+ . In ESR detected . In the ous data \u201352). All2(tmd)/Ph2I+/NVK initiating system upon a laser diode at 532 nm is depicted in 3/Ph2I+/NVK leads to a low conversion (<20%) in full agreement with the low absorption of Ir(ppy)3 under green lights (see The polymerization profile for the ring-opening polymerization of EPOX using a Ir(piq)ghts see . Remarkaghts see , is in a2(tmd)/MDEA/R\u2013Br is very efficient (2(tmd)/MDEA (The performance of the new proposed PIC for a reductive cycle in combination with an amine (MDEA) and an alkyl halide to initiate a radical polymerization has been also checked. In full agreement with fficient , curve 2md)/MDEA , curve 12(tmd)). This allows the design of multicomponent photoinitiating systems based on an excellent Ir complex working at \u03bb > 500 nm, a goal that was never achieved before using other available Ir derivatives 3). The development of new catalytic methodologies still remains a huge challenge.Photoinitiator catalysts PICs appear as a new class of initiating systems usable in different photopolymerization reactions: FRP, CP and FRPCP. The associated systems are characterized by an outstanding photosensitivity; the catalytic pathways ensure a regeneration of the PIC and avoid any lost of reactivity upon irradiation. A bleaching of the sensitizer can be observed in excess of oxidation or reduction agent. In this case the polymerization of thick samples can be carried out. With this bleaching, colorless coatings can also be obtained. The search for new PICs and/or new redox agents might be of high interest as depicted by the present proposal of a new PIC (Ir(piq)"} +{"text": "The shape of nuclei in many adherent cultured cells approximates an oblate ellipsoid, with contralateral flattened surfaces facing the culture plate or the medium. Observations of cultured cell nuclei from orthogonal perspectives revealed that nucleoporin p62 (NUP62) and nucleoporin 214 (NUP214) are differentially distributed between nuclear pore complexes on the flattened surfaces and peripheral rim of the nucleus. High resolution stimulated emission depletion (STED) immunofluorescence microscopy resolved individual NPCs, and suggested both heterogeneity and microheterogeneity in NUP62 and NUP214 immunolabeling among in NPC populations. Similar to nuclear domains and interphase chromosome territories, architectural diversity and spatial patterning of NPCs may be an intrinsic property of the nucleus that is linked to the functions and organization of underlying chromatin. Nuclear pore complexes (NPC) assemble in the nuclear envelope (NE) from more than 30 different proteins, and are organized into multiprotein subcomplexes. Multiples of eight units of each subcomplex assemble into the modular structures of the pore, which include a symmetric pair of core inner ring complexes, an asymmetric pair of annular rings on the cytoplasmic and nuclear faces, and asymmetric filamentous structures projecting into the nucleoplasm and cytoplasm Located symmetrically on both the cytoplasmic and nuclear faces of the NPC, NUP155 interacts with Gle1 and nucleoporin pCG1 to mediate mRNA transport Export of mRNA from the nucleus is a complex process that in yeast and probably in metazoans is linked to mRNA processing. Export proteins that interact with the NPC and mediate transport recognize adaptors that bind maturing RNA, and many of these adaptors are directly involved in RNA processing are likely to represent NPCs, whereas intranuclear signals, by definition, would not arise from NPCs and may represent nucleoporins involved in functions other than nucleocytoplasmic transport Western blot analyses revealed that knockdown of NUP62 or NUP214 to \u223c25% of control levels resulted from treatment of TOV112D cells . In HEK293 cells, optical sections through the central plane of the nucleus display overlapping punctate NUP62 and NUP214 signals on the rim of the nucleus, a distribution considered typical for NPC immunolabeling . The NUPFurther studies of NUP62 and NUP214 immunolabeling in TOV112D cells were performed using expression constructs for V5 epitope-tagged NUP62 or NUP214 cDNAs. Plasmids were transfected into TOV112D cells, and processed for immunofluorescence microscopy 72 hours later using NUP62, NUP214, and/or V5 epitope tag antibodies. Observations were restricted to pairs of post-mitotic cells. As NPCs and the nuclear envelope are disassembled during mitosis then reassembled during abscission and early G1 phase, observation of post-mitotic pairs of transfected cells assures that the V5 epitope-tagged forms of NUP62 and NUP214 have distributed into the total NPC and nucleoporin pools. Cells transfected with V5 epitope-tagged NUP62 or NUP214 expression constructs were analyzed by immunofluorescence with V5 epitope antibody and NUP62 or NUP214 antibodies, respectively. Extensive co-localization of NUP62 or NUP214 immunolabeling with V5 epitope tag immunolabeling was observed in transfected TOV112D cells , confirmThe V5 epitope tags were inserted at the N-termini of NUP62 and NUP214. The NUP62 antibody recognizes an epitope within residues 24\u2013178 and the NUP214 antibody recognizes epitopes between residues 1250\u20131300 of the human form of the protein. The nucleoporin immunolabeling patterns generated by native and epitope tag antibodies overlapped extensively, suggesting that immunofluorescence patterns generated by the native NUP62 or NUP214 antibodies reflect the presence or absence of nucleoporins and not post-translational modifications that alter antibody binding . The TOVImmunoblot analyses revealed+/NUP214\u2212, NUP62\u2212/NUP214+, and NUP62+/NUP214+ immunolabeled NPCs varied between cultured cell types. Ovarian carcinoma TOV112D cells and COS7 cells displayed two major populations of NPCs: peripheral rim NUP62+/NUP214\u2212 and contralateral surface NUP62\u2212/NUP214+ NPCs, with few NUP62+/NUP214+ NPCs observed , binding of both NUP62 and NUP214 antibodies to the same NPC is not precluded if both nucleoporins are present. This supports the interpretation that differences in immunolabeling represent differences in the distribution of NUP62 and NUP214 among NPCs rather than variations in the accessibility of the epitopes. The observed NPC patterns of the cultured cell types were consistent among most of the cells in a proliferating culture .T cells combined with scanning confocal microscopy resolved immunofluorescent images of individual NPCs. The S12 neuroblastoma cell line was chosen for this analysis as it displays each of the three permutations of NUP62/NUP214 immunolabeled NPC types. Individual optical sections from the surfaces facing the culture plate or the medium and the central plane of the S12 nucleus revealed both common and disparate immunolabeled patterns for NUP62 and NUP214 . Punctat+/NUP214\u2212 and NUP62\u2212/NUP214+ NPC populations, NUP62>NUP214 and NUP6280,000 years ago. Data from velocity-dependent microfracture features, diagnostic damage patterns, and artifact shape reported here indicate that pointed stone artifacts from Ethiopia were used as projectile weapons (in the form of hafted javelin tips) as early as >279,000 years ago. In combination with the existing archaeological, fossil and genetic evidence, these data isolate eastern Africa as a source of modern cultures and biology.Projectile weapons (i.e. those delivered from a distance) enhanced prehistoric hunting efficiency by enabling higher impact delivery and hunting of a broader range of animals while reducing confrontations with dangerous prey species. Projectiles therefore provided a significant advantage over thrusting spears. Composite projectile technologies are considered indicative of complex behavior and pivotal to the successful spread of Homo sapiens to expand out of Africa and outcompete Neanderthals A key component in prehistoric subsistence strategies, the invention of projectile weapons was a decisive advance over the thrusting spear \u223c400 ka were likely used in hunting large game At Kathu Pan, in South Africa, Middle Pleistocene hominins made hafted stone-tipped hunting spears \u223c500 thousand years ago (ka); these were, however, not projectiles but hand-delivered thrusting weapons ca. 100-50 ka The identification of prehistoric projectile weaponry has been largely inferential. Paleolithic archaeologists suggest that mechanically-projected weapons, such as the bow-and-arrow, originated among modern humans in Africa Homo as early as >279 ka. Specifically, we apply the velocity-dependent microfracture approach to Middle- and Late Pleistocene pointed artifacts to reliably identify the mode(s) of weapon delivery.Here we report on multiple independent lines of evidence that strongly indicate that pointed obsidian artifacts recovered from sites in the Gademotta Formation (Fm.) of the Main Ethiopian Rift represenMicrofracture features, such as fracture wings , are velocity-dependent ripple marks created when a crack force encounters intrinsic imperfections (such as bubble pores) in materials like obsidian 40Ar/39Ar methods to >279\u00b12 ka 40Ar/39Ar results reported here now provide a tighter minimum age constraint for these sites. The youngest occupation (ETH-72-6) is constrained to between 185\u00b15 The Gademotta Fm. archaeological site complex is located in the flanks of an ancient collapsed caldera in the central sector of the Main Ethiopian Rift . It cont40Ar/39Ar analyses were conducted on tephra samples collected from fine ash (Trench1-Step1) as well as pumice deposits (Trench1-Step2) of Unit 12 in the type-site were used to record the three-dimensional co-ordinates of each The Gademotta artifacts are made almost exclusively on obsidian. Geochemical provenancing establishes that the obsidian used for artifact manufacture at Gademotta came from the nearby (<2.5 km distant) obsidian source at Kulkuletti/Worja All 226 convergent artifacts (171 pointed tools and 95 point fragments) were examined; 141 were found to bear fracture patterns potentially attributable to impact from their use as weapons. All of these pieces are made on obsidian. Microfracture analysis assessed velocity-dependent fracture features to identify use-related precursory loading Fracture velocity analyses of pointed artifacts identified and measured plane fracture wings. Fracture wings (FWs) are V-shaped, with their apex pointing toward the direction of fracture propagation Fracture velocity analysis involved multiple stages: i) determination of the physical properties of the obsidian raw material exploited by prehistoric inhabitants of the Gademotta sites, as described below; ii) microscopic investigation of fracture surfaces on pointed pieces; iii) capturing of microfracture features in photomicrographs; iv) measurement of dimensions of microfracture features; v) calculation of instantaneous fracture velocity.n\u200a=\u200a32) were collected from the Kulkuletti/Worja primary source E) and Poisson\u2019s Ratio (v) for the Kulkuletti/Worja obsidian were determined by the pulse method using the NDT James Instruments MK IV ultrasonic transducer in the National Museum of Ethiopia, Addis Ababa. These properties were used to calculate the Modulus of Rigidity (G). The distortional wave velocity (2C) of the Worja obsidian was computed from G and density (\u03c1) of the Kulkuletti/Worja obsidian. Density was measured in the Concrete Materials and Structural Integrity Research Unit (CoMSIRU) of the Department of Civil Engineering, University of Cape Town. The following formulae and procedures detailed in Hutchings Non-artifactual obsidian samples (C) was calculated from the angles of FWs (C\u200a=\u200acos ([\u03c8/2]* C2); where \u03c8 is the angle of divergence of a plane fracture wing. Measurements of angle of divergence were conducted on digital versions of photomicrographs using the built-in package on the Keyence VHX-600 (3CCD) digital microscope housed in the NME, and independently using the external software packages MB Ruler 4.0 (http://markus-bader.de/MB-Ruler) and ImageJ 1.440 (http://imagej.nih.gov/ij). Instantaneous fracture velocity magnification on binocular reflected light microscope were employed infrequently. Macroscopic fracture types most commonly considered diagnostic of impact from use of pointed pieces as weapon tips include: i) burin-like fractures; ii) flute-like fractures; iii) transverse fractures with terminations other than snaps that were inflicted after the artifact was retouched; iv) bifacial spin-off fractures; v) unifacial spin-off fractures with a fracture length of >6 mm potential of pointed artifacts to have served as tips of projectile weapons Artifact morphology is one important variable that prehistoric hunters optimized to achieve the desired aerodynamic qualities and penetrative abilities of hafted points TCSA and TCSP were calculated following the methods detailed by Shea For TCSP, values from the more restrictive measure of triangular, rather than rhomboidal, cross-section were used, as recommended by Sisk and Shea 40Ar/39Ar analysis on samples Trench1step1 and Trench2step2 yielded analytically indistinguishable results, with a combined isochron age of 260\u00b17 ka . This evidence for artifact modifications to facilitate hafting supports previous inferences from use-wear analyses in which microscopic striations on the surfaces of Gademotta points were interpreted as the result of hafting Macroscopic edge damage analysis shows that just over 81% (13 out of 16) of the pointed pieces with impact-induced microfractures also display macrofeatures referred to as diagnostic impact fractures (DIFs) n\u200a=\u200a113 from all six sites in the Formation) are statistically indistinguishable p\u200a=\u200a0.6598 for TCSA; \u22120.1211; p\u200a=\u200a0.9037 for TCSP; n\u200a=\u200a71) from a substantially younger (90 ka) MSA context at Klasies River main site (KRM), South Africa Morphometric analyses show that both the TCSA and TCSP of the GDM point assemblage relative probability and (B) inverse isochron of single crystal total fusion analyses for sanidines for samples T1S1, T1S2 (in red), and combined results from both samples. Xenocrysts are shown in pink on A, and are excluded from age calculations; they are not included on B.(TIF)Click here for additional data file.Figure S2Pictures showing (A) a fracture surface containing FWs sampled for analysis from a locus at 34.7% of the crack length; (B) a photomicrograph of plane FWs; and (C) the measurement of angle of divergence of a prominent FW.(TIF)Click here for additional data file.Table S1Material properties of the Kulkuletti/Worja obsidian. Data were collected via the pulse method. The ultrasonic transducer was set to 1 pulse per second for all measurements. The average E value was converted to Newton/m2, yielding a value of 8.9425e+10 N/m2, cf. 3.(PDF)Click here for additional data file.Data S1Full Ar data for Trench1step1 and Trench1step2 samples.(XLS)Click here for additional data file."} +{"text": "LGE imaging of left atrial scar is promising and can detect pre/post ablation procedures however further quality control required to accurately depict lesion distributionWe tested the hypothesis that cardiovascular magnetic resonance (CMR) imaging can reliably distinguish the presence or absence of ablation lesions by blinded analysis of pre and post ablation imaging.Consecutive patients with paroxysmal AF in a randomised study comparing pulmonary vein isolation by wide area circumferential radiofrequency ablation (WACA) to ostial ablation with a cryo-balloon (CRYO) underwent CMR late gadolinium enhancement (LGE) imaging pre- and 3 months post ablation. Imaging was anonymized for blinded analysis of (1) LGE images, and (2) a 3D fusion image with LGE projected onto a segmented LA surface . Scans were categorised using both assessment techniques separately as pre or post ablation, and if post ablation, whether lesions were in an ostial or WACA distribution.LGE imaging was performed in 50 patients . Sensitivity and specificity for detection of ablation lesions was 60% and 96% on LGE imaging, or 88% and 82% respectively on 3D fusion imaging. Detection of WACA and CRYO lesion sets were correct in 14/24 and 16/26 respectively on LGE imaging, or 21/24 and 23/26 on 3D fusion imaging. Assessment of lesion distribution was correct for 13/50 from LGE images (4/24 for WACA and 9/26 for CRYO) and 26/50 from 3D fusion images .LGE imaging of atrial scar after ablation therapy is feasible. The technique still needs quality control standards established in order to determine the appropriate SD setting and to judge whether lack of peri-ostial scar is due to a sub-optimal scan or myocardial recoveryNational Institute of Health Research (UK)."} +{"text": "Continuous interscalene nerve blockade (CISNB) has been shown to reduce postoperative pain and morphine usage after a wide range of shoulder operations (1&2). Recent trends to minimally invasive shoulder surgery are expected to be associated with shorter postoperative stays. This study compared the length of stay using CISNB as the primary postoperative analgesia with other forms of analgesia and measured the time from end of surgery to discharge.Following ethics committee and institutional approval from the participating hospital , the investigator examined the medical records of all patients having minimally invasive shoulder surgery undertaken by the one surgeon. Included were shoulder acromioplasty, shoulder capsulotomy, superior labral anterior posterior (SLAP) repair and rotator cuff repair (mini-open). Exclusions were patients under 16 years, ASA status greater than 3, bilateral surgery and emergency cases.There was no difference between groups with median hospital length of stay from the end of surgery to discharge being 24.4 hours (quartiles 20.7-42.1) for the CISNB (A) group compared with 24.0 hours (18.8-42) for the \u201cother analgesia \u201c(B) group which included parenteral morphine alone or in combination with single shot interscalene nerve block or intra-articular local anaesthetic. Secondary observed outcomes included opioid use on the ward in morphine equivalents with CISNB patients requiring a median dose of 2mg (quartiles 0-9) compared with \u201cother analgesia\u201d needing 6mg (1-16) p=<0.01.For minimally invasive shoulder surgery the presence of CISNB is not associated with shortened hospital stay despite requiring less morphine equivalents. Further analysis of patients with stays over 48 hours showed rotator cuff repairs and age over 65 may be associated with longer stay however this study was underpowered to prove this association."} +{"text": "Progression of scoliosis may lead to self-esteem issues, pain, respiratory complications and limited function. There is a paucity of information regarding how physical therapists manage adolescent idiopathic scoliosis (AIS).The aim of this study was to determine the objectives, treatments and outcomes addressed by physiotherapists in the non-operative management of AIS.A web survey was emailed to 1599 outpatient physiotherapists in Alberta identified from the College of Physical Therapists of Alberta\u2019s registry in 2009. The 30 question survey was adapted from previous back pain studies [Only 15% of all patients with AIS were referred to physiotherapy with a mean age of 16 \u00b1 3 years, and a mean Cobb angle of 26 \u00b1 15 degrees. The response rate from therapist was 11.9%,after 2 reminders with valid responses from 147 therapists. The top objectives pursued by physiotherapists in Alberta were pain reduction (80%), stopping curve progression (57%) and improving body image (45%). Therapists with 0-10 years of experience ranked pain reduction and body image improvements significantly higher than therapists with 10-20 or over 20 years of experience. No therapist reported using scoliosis specific exercises. Stabilization exercises (76%), non-scoliosis specific postural approaches (73%), and mobilizations (55%) were the highest ranked treatment methods used, with mobilizations being used significantly more frequently in rural settings. The primary outcomes documented by physical therapists in Alberta were pain (75%), subjective posture observation (73%), and range of motion (69%).The objectives, treatments, and outcomes pursued by Alberta\u2019s physiotherapists while managing AIS are variable, depending on experience and practice settings. Alberta practices did not fully match published recommendations supporting quality of life as primary therapy objective and the"} +{"text": "Since publication of our article , we obse\"Results\" section of the Abstract, the end of the first sentence should read as follows:In the \"after five years in young adult males (p for trend \u2264 .02).\"In the same section, the next sentence should read:\"The positive relationship between maternal encouragement and MVPA approached significance among high-school aged females (p for trend = .06), and a positive relationship between parental encouragement and MVPA approached significance among high school-aged males (p for trend = .05).\"Results\" section of the article, the second sentence should read:In the \"At Time 2, participants reported engaging in MVPA between 3.9 and 6.8 hours perweek and reported watching TV/video between 17.1 and 19.2 hours per week ...\"Physical activity section of the Results:In the The end of the second sentence should read:\"years later (p for trend \u2264 .02).\"The middle of the third sentence should read:\" engaged in MVPA 2.2 more hours...\"The middle of the fourth sentence should read:\"participating in 5.1 hours of MVPA per week compared to 7.3 hours per week...\"The sixth sentence should read:\"among younger and older males (p for trend \u2264 .01)\"and... \"among older females (p for trend = .05)\" should not be present.The end of the seventh sentence should read:\"among older males remained (p for trend \u2264 02)\"The end of the eighth sentence should read:\"approached significance (p for trend = .05).\""} +{"text": "Previous genome-wide expression studies have highlighted distinct gene expression patterns in inflammatory bowel disease (IBD) compared to control samples, but the interpretation of these studies has been limited by sample heterogeneity with respect to disease phenotype, disease activity, and anatomic sites. To further improve molecular classification of inflammatory bowel disease phenotypes we focused on a single anatomic site, the disease unaffected proximal ileal margin of resected ileum, and three phenotypes that were unlikely to overlap: ileal Crohn's disease , ulcerative colitis (UC), and control patients without IBD. Whole human genome (Agilent) expression profiling was conducted on two independent sets of disease-unaffected ileal samples collected from the proximal margin of resected ileum. Set 1 was used as the training set and Set 2 was subsequently collected as an independent test set . We compared the 17 gene signatures selected by four different feature-selection methods to distinguish ileal CD phenotype with non-CD phenotype. The four methods yielded different but overlapping solutions that were highly discriminating. All four of these methods selected FOLH1 as a common feature. This gene is an established biomarker for prostate cancer, but has not previously been associated with Crohn's disease. Immunohistochemical staining confirmed increased expression of FOLH1 in the ileal epithelium. These results provide evidence for convergent molecular abnormalities in the macroscopically disease unaffected proximal margin of resected ileum from ileal CD subjects. Transcriptomic analyses have highlighted differences in intestinal gene expression patterns between samples collected from patients with inflammatory bowel disease (IBD) compared to control patients without inflammatory bowel disease MUC1, DUOX2 and DMBT1 expression and decreased expression of C4orf7 (follicular dendritic cell secreted peptide) was confirmed by reverse transcriptase polymerase chain reaction of 18 ileal CD and 9 control non-IBD samples. We found that these alterations in gene expression were independent of NOD2 genotype We have previously examined genome wide expression profiles in the disease unaffected proximal margin of resected ileum collected from 4 patients with Crohn's disease of terminal ileum undergoing initial ileocolic resection with that of 4 control non-IBD patients undergoing initial right hemicolectomy or total colectomy To better define the molecular characteristics of the ileal CD phenotype, we applied four different feature selection methods to select 17-gene signatures that would distinguish samples of the proximal disease unaffected proximal margin of ileum that were resected from individuals with ileal CD phenotype, from samples collected from non-CD phenotype (both non-IBD and ulcerative colitis patients) to a training set composed of 99 expression profiles. We then tested these features in an independently collected test set of 30 expression profiles.disease-unaffected was based on the macroscopic appearance of the ileal mucosa and the surgical pathology report of adjacent ileal biopsies . The clinical information and samples were collected as previously described This study was approved by the Washington University-St. Louis and Stony Brook University Institutional Review Boards. Ileal CD patients undergoing ileocolic resection, UC patients undergoing total colectomy and Control non-IBD patients undergoing either right hemicolectomy or total colectomy were prospectively enrolled in a consecutive fashion by the Washington University Digestive Diseases Research Core Center Tissue Procurement Facility to donate surgically resected tissue samples between September 2005 and December 2010. A subset of 8 of the 99 expression profiles generated from samples collected between September 2005 and February 2010 in the training set were previously reported http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE24287).Total RNA was extracted from the tissue samples using TRI Reagent\u00ae according to the manufacturer's recommendation, and RNA quality was assessed using an Agilent 2100 Bioanalyzer et alTwo-class unpaired significance analysis of microarrays (SAM) was performed on 25,756 probes in the training set as previously described by Tusher Folate hydrolase 1 (FOLH1), also termed prostate specific membrane antigen (PSMA) C. difficile was more prevalent among UC patients than ileal CD or control non-IBD patients The patients included in this study were predominantly white. As shown in DMBT1 expression level was confirmed to be significantly increased, while that of C4orf7 was confirmed to be significantly decreased in the disease unaffected proximal margin of ileum resected from ileal CD patients compared to nonIBD Control and UC patients MUC1 and DUOX2 expression was increased relative to Control samples. However because MUC1 and DUOX2 expression was also increased in UC compared to nonIBD Control samples, these genes were not selected in this two-class unpaired SAM comparing ileal CD and non-CD (UC and Control).Because a large amount of variability can be introduced in the fold change for low intensity signals, the threshold for gene filtering was selected to be twice the background, resulting in a total of 25,676 gene-probes FOLH1) gene was selected by all four feature selection methods. Three known genes, TLR4 interactor with leucine rich repeats (TRIL), Niemann-Pick disease, type C1, gene-like 1 (NPC1L1), and C4orf7 also termed follicular dendritic cell secreted protein were selected by three of the four methods. Six known genes were selected by two of four methods, BCL2-associated X protein (BAX), cytochrome P 450, family 26, subfamily B, polypeptide 1 (CYP26B1), nephronectin (NPNT), protein phosphatase 1, regulatory (inhibitor) subunit 14A (PPP1R14A), family with sequence similarity129, member C (FAM129C) also termed B-cell novel protein 1 (BCNP1), cathelicidin antimicrobial peptide (CAMP), chemokine (C-C motif) ligand 23 (CCL23). We repeated our analysis using data excluding the C. difficile positive samples. FOLH1 is still the only gene probe selected by all four feature selection methods and it is still ranked prominently by all four classifiers 2nd, 1,st, 1st and 4th by PAM, RF, LASSO and Boosting, respectively). In addition, ten out of 12 genes selected by two or more feature methods based on data without C. difficile positive samples overlap with those selected using data including the C. difficile positive samples. Meanwhile, the Bossting method still features the highest classification accuracy at 89.90% and 86.96% for data with and without the C. difficile positive samples, respectively. All these observations indicate that our method was not skewed by the C. difficile toxin factor.Four feature subset selection methods Boosting , were apMajority vote based on the median score of seven classifier tools see was usedFOLH1) gene was identified as a \u201chub\u201d gene that has significantly non-zero partial correlations to 12 of the other 16 gene biomarkers that we noted previously to be upregulated in ileal CD with control non-IBD subjects were not selected in the current study because these genes were also upregulated in UC compared to control samples In this study, we took a statistical approach to identify ileal gene biomarkers associated with ileal CD phenotype compared to non-CD (UC and control). Some of the genes samples (number of probes\u200a=\u200a269). B. Downregulated probes in 47 ileal CD compared to 52 non-CD samples .(DOCX)Click here for additional data file.Table S2bolded.Union of differentially expressed Agilent gene probes selected by four feature subset selection methods: Boosting, PAM, Random Forest (RF) and LASSO. A. Upregulated probes B. Downregulated probes The 17 genes selected by the boosting method are (DOCX)Click here for additional data file.Table S3AUC values of empirical ROC curves, AUC values and 95% confidence interval (C.I.) of smoothed ROC curves.(DOCX)Click here for additional data file."} +{"text": "Stimulation of neurons with brain-derived neurotrophic factor (BDNF) results in robust induction of SORLA, an intracellular sorting receptor of the VPS10P domain receptor gene family. However, the relevance of SORLA for BDNF-induced neuronal responses has not previously been investigated. We now demonstrate that SORLA is a sorting factor for the tropomyosin-related kinase receptor B (TrkB) that facilitates trafficking of this BDNF receptor between synaptic plasma membranes, post-synaptic densities, and cell soma, a step critical for neuronal signal transduction. Loss of SORLA expression results in impaired neuritic transport of TrkB and in blunted response to BDNF in primary neurons; and it aggravates neuromotoric deficits caused by low BDNF activity in a mouse model of Huntington\u2019s disease. Thus, our studies revealed a key role for SORLA in mediating BDNF trophic signaling by regulating the intracellular location of TrkB. Sorting protein-related receptor containing LDLR class A repeats (SORLA) is a member of the VPS10P domain receptor gene family, a class of sorting proteins expressed in the mammalian nervous system Sorl1 (the gene encoding SORLA) is a downstream target of brain-derived neurotrophic factor (BDNF), a growth factor that signals through tropomyosin-related kinase receptor (TrkB) to promote neuronal survival Sorl1 transcription 10-fold whereas absence of BDNF activity in mouse models with genetic (\u2212/\u2212Bdnf ) or disease-related loss of BDNF in the brain results in impaired SORLA expression and activity Recently, the relevance of SORLA as a neuroprotective factor received independent support by findings that Using proteomics approaches combined with functional studies in cultured neurons and in mouse models of HD, we now identified SORLA as novel sorting factor for TrkB that facilitates transport of this BDNF receptor along neurites to enhance BDNF signaling. Thus, BDNF-mediated induction of SORLA expression may represent a cellular mechanism to potentiate trophic signals, a pathway potentially disrupted in HD.All experiments performed with mice were conducted according to the guidelines of the German Animal Welfare Law. The study was approved by the State Office of Health and Social Affairs Berlin .\u2212/\u2212Sorl1Generation of the Huntington\u2019s disease mouse model (B6C3-Tg(HD82Gln)81Dbo/J) carrying a transgenic huntingtin gene with 82 CAG repeats Antisera have been obtained from Cell Signaling Technology or Santa Cruz Biotechnology. Plasmids coding for TrkB-mCherry, TrkB-EGFP, and SORLA-EGFP have been generated by introducing corresponding cDNA into the pCI Mammalian Expression Vector (Promega) and pcDNA3 vector (Invitrogen), respectively.Primary cortical neurons were prepared from newborn Balb/c mice of either sex at postnatal day 1. Cortices were dissociated in papain (1 hour at 37\u00b0C) and cultured on poly-D-lysine/collagen coated culture dishes. The neurons were cultured for 5 days in Neurobasal-A medium (Gibco) including B27 supplement (Sigma), and GlutaMAX (Invitrogen) as previously described Neurons were treated with BDNF or medium only (control) for 20 minutes or 48 hours (BDNF-dependent proteome changes) by replacing half of the culture medium with fresh medium. For proteome analyses the cells were harvested in ice cold PBS and cell pellets were frozen immediately in liquid nitrogen. Six individual samples of each treated and control cells were collected (n\u200a=\u200a6).For proteome analyses, the cells were harvested in ice cold PBS. Six individual samples of each treated and control cells were collected (n\u200a=\u200a6). Protein extracts were prepared from frozen cell pellets and separated by large-gel 2D-PAGE technique as described For protein identification, 1200 \u00b5g protein extract was each separated on a 2-D gel and stained with a MS-compatible silver staining protocol Brain tissue was homogenized using a glass-Teflon Potter S homogenizer with 10 strokes at 900 rpm on ice. Homogenates were cleared through differential centrifugation. Synaptosomes were subjected to osmotic shock and put on a discontinous sucrose gradient . Further enrichment of different fractions was achieved through additional steps of centrifugation as described +/+Sorl1) and 228 vesicles (\u2212/\u2212Sorl1) in 13\u201316 neurons per genotype were analyzed (three independent experiments). Vesicle movement was quantified by generation of kymostacks using ImageJ KymoToolbox plugin .SH-SY5Y cells +/+Sorl1) or genetically deficient for SORLA (\u2212/\u2212Sorl1) to BDNF stimulation. Following the experimental conditions of earlier studies on chronic Sorl1 induction +/+Sorl1 or \u2212/\u2212Sorl1 mice were treated for 48 hours with 150 ng/ml BDNF or medium only (control). The global response and Akt pathways neurons . In cont neurons .Taken together, the analysis of signal cascade activation identified a blunted response to BNDF in neurons lacking SORLA despite proper expression of TrkB.Functional expression of Trk receptors in neurons involves extensive axonal transport processes whereby these receptor molecules shuttle between cell body and synapses compared to parental control cells (SY5Y) lacking the protein . A shiftTogether, our results demonstrate that SORLA interacts with TrkB to determine TrkB surface expression. Loss of SORLA results in accumulation of TrkB at the cell surface. This is in line with our previous result demonstrating blunted signal cascade activation as this process depends on internalization and transport of activated TrkB receptors to intracellular compartments.+/+Sorl1 and \u2212/\u2212Sorl1 primary neurons.Altered subcellular localization of TrkB and corresponding changes in TrkB signaling in SORLA-deficient neurons may be caused by defects in either anterograde, retrograde, or bi-directional transport of the BDNF receptor. Accordingly, we applied time-lapse microscopy to measure movement of vesicles containing a fluorescently labeled fusion protein of TrkB with EGFP in As shown in Taken together, analysis of TrkB localization in cells and in the brain as well as evaluation of TrkB trafficking in transfected primary neurons argued for a role of SORLA in transport of TrkB.\u2212/\u2212Sorl1) and compared them to control HD82 animals (HD82x+/+Sorl1).In HD, levels of BDNF are distinctly reduced in the striatum of affected individuals \u2212/\u2212Sorl1 mice displayed pronounced hind limb clasping compared with HD82 control litter mates (HD82x+/+Sorl1) when suspended by their tail (\u2212/\u2212Sorl1 failed to do so and performed significantly poorer on day 3 and 4 (\u2212/\u2212Sorl1 with wild-type controls (+/+Sorl1) , indicatSorl1 that activates receptor gene transcription through the ERK pathway. Induction of Sorl1 transcription results in a robust increase in SORLA protein expression and activity in cultured neurons and in the mouse brain in vivoUsing screening approaches in primary neurons, we previously identified BDNF as a major inducer of \u2212/\u2212Sorl1 neurons, results in enhanced accumulation of TrkB at the synaptic plasma membrane and in depletion from the post-synaptic density compartment (To elicit its biological effects, BDNF signals must be conveyed over long distances from the nerve terminal to the cell body partment . Also, apartment . SeveralSorl1 but not the gene encoding sortilin is induced by BDNF in neurons Sorl1A similar function as for SORLA in trafficking of TrkB has been documented for sortilin, a related receptor,of the VPS10P domain receptor family BDNF signaling has long been recognized as an autocrine mechanism whereby adult sensory neurons secrete BDNF to sustain their own survival signals"} +{"text": "An image analysis method was developed to obtain a 3D map of the pyruvate metabolites distribution in the LV in hyperpolarised 13C MRI. The obtained polar maps follow the standardized LV AHA segmentation, allowing reproducible and standardized LV segmental analysis.MRI with hyperpolarised 13C represents a promising modality for dynamic in vivo spectroscopy and could provide a unique opportunity for non invasive assessment of regional cardiac metabolism . The aimSeven male pigs 38\u00b12 kg) were imaged on a 3T MRI scanner (GE Excite Hdx) with a 13C birdcage coil. 13-C-pyruvate (20mL of 230mM) was hyperpolarized using a Dynamic Nuclear Polarization technique (Hypersense) and injected in pigs at rest and during occlusion of the left anterior descending coronary artery. Anatomical reference images were acquired by a standard SSFP sequence. Metabolic information were obtained using a volumetric IDEAL spiral CSI sequence prescrib8\u00b12 kg weDifferences in metabolite signal among acquisitions were normalized by assessing segmental variation as percent difference between segmental values and global LV value. To assess the difference between basal and occlusion conditions, the mismatch between two segmental variation maps was defined as percent difference. Fig. MRI with hyperpolarised 13C allows mapping the pyruvate metabolites distribution in the LV following the standardized AHA segmentation. Macroscopic changes in metabolites concentration due to coronary occlusion are consistently detected. Further studies are needed to characterize the sensitivity of the method to fine variations of regional metabolite concentration.No specific funding was received for this study."} +{"text": "HIV rapid tests are widely used in resource limited settings. Dried blood spots on filter paper are an easy way to collect and transport blood samples from a remote area to a testing facility at ambient temperatures. Our objective was to evaluate the technical possibility of testing antibodies to HIV from filter paper using HIV rapid tests kits.Dried blood spots on filter paper were obtained from 408 female sex workers (FSW), 136 Men Sex with Men (MSM) and 50 Intravenous Drug Users (IDU). Antibodies were eluted from DBS disks in 200 \u00b5l of sterile Phosphate Buffer Saline (PBS) by agitating overnight at 2-8\u00b0 C (refrigerator) for minimum 16 hours and tested for anti-HIV antibody using two ELISA . As per the guidelines of the National AIDS Control Organization Sentinel Surveillance of HIV, the reactive samples on the first ELISA are retested using a 2nd ELISA kit. We assessed all the HIV positive and HIV negative DBS samples using 3 HIV rapid tests kits procedures were followed according to the manufacturer's instructions.Of the 594 DBS samples, 22 (3.69%) samples were positive for HIV antibodies using ELISA kits. Among HIV positive DBS samples, 3.67% (15/408) were FSW's, 2.95% (4/136) were MSM and 6% (3/50) were IDU's. All 22 DBS were positive in both ELISA kits and the sensitivity and specificity of both kits were 100%, whereas Determine found 20 HIV positives, Combaids 21 samples Positive and EIAComb found all 22 HIV positive giving sensitivity was 90.91%, 95.45%, and 100% respectively and specificity was 100%.This study demonstrates and confirms the usefulness and feasibility of the filter paper blood collection method for testing of HIV antibodies. DBS can be used with HIV rapid test devices. Using the latest and advanced techniques applied in rapid test technologies, DBS may provide a unique way to conduct sero epidemiological surveys in resource limited settings."} +{"text": "During the 15-min recovery phase the plasma nitrite, AIx and MDA values remained altered.Breathing of normobaric 100% oxygen during 30 min caused an increase of P\u2022NO bioavailability due to elevated and sustained oxidative stress.This study suggests that the underlying mechanism of hyperoxia-induced vasoconstriction may result from reduced"} +{"text": "The degree and quantification of contractile dysfunction and myocardial scar in patients after acute myocardial infarction (AMI) has important prognostic implications. Myocardial deformation parameters like strain and strain rate have been shown to be more sensitive markers for contractile dysfunction than standard 2-D parameters.We sought to investigate whether strain and strain rate imaging, assessed by a novel non-invasive post-processing feature tracking algorithm (FTI) on pre-acquired regular CMR SSFP images, would allow quantification of regional left ventricular (LV) function and its relation to degrees of infarct trans-murality in patients after AMI.Cardiac magnetic resonance (CMR) imaging was performed on a 1.5T whole body MRI scanner (Philips Achieva) in 44 patients (mean age: 56 \u00b1 12) 3 \u00b1 1 days after successfully reperfused first ST segment elevation AMI. Peak systolic circumferential strain and strain rate were measured on standard CMR SSFP cine sequences and afterwards related to the trans-mural extent of myocardial infarction determined by contrast enhanced CMR (CE-CMR) 10 minutes after i.v. injection of 0.2mmol/kg Gd-DTPA (Magnevist). The feature tracking algorithm is a two dimensional deformation analysis of the myocardium that was originally designed for echocardiographic image analysis, which has now been transferred to CMR SSFP sequences without the need for additional scans.Cut-off peak systolic circumferential strain values of -21% differentiated patients with global infarct sizes \u226520g from patients with smaller infarct sizes providing sensitivities and specificities of 94% and 74%. An infarcted myocardium with a segmental infarct size of \u226550% was differentiated from non-transmural infarcted segments with a cut-off peak systolic circumferential strain value of >-14% with a sensitivity of 92% and a specificity of 86%. Peak negative circumferential strain correlated closely with the quantitatively determined infarct size on a global and segmental level .FTI provides a rapid and objective quantification of global and regional myocardial function and allows the discrimination between different degrees of trans-murality in patients after AMI.none"} +{"text": "Ciliogenesis involves coordinated assembly of a microtubule-based axoneme from the mother centriole and vesicular membrane transport and fusion forming a ciliary membrane around the developing axoneme. We, and others have reported that a Rab11-Rab8 cascade functions in ciliogenesis. Using live high-resolution fluorescence microscopy imaging we show that ciliary membrane assembly proceeds following Rabin8 (a Rab8 activator) binding to Rab11 membranes. Rabin8 transport via Rab11 vesicles to the centrosome is observed resulting in localized activation of Rab8 and leads to initiation of ciliary membrane assembly. Using proteomics approaches, we have discovered that Rabin8 binds to the TRAPPII tethering complex and find that this interaction is important for Rabin8 centrosomal targeting during ciliogenesis. Our work suggests that Rabin8 membrane transport is a highly regulated process controlled by serum-dependent and serum-independent signaling. Interestingly, following ciliogenesis Rabin8 centrosomal localization is lost resulting in reduced Rab8 activation at the ciliary membrane. This finding along with a previous report describing Rabin8 association with Bardet-Biedl syndrome (BBS) proteins has led us to hypothesize that regulation of centrosomal Rabin8 levels is important for establishing the length of primary cilium, an important factor in ciliary signaling. Finally, we describe the discovery of additional factors associated with the Rab11-Rab8 trafficking pathway that function in organizing membrane structure during ciliogenesis."} +{"text": "The present study has been conducted to investigate whether phyllanthus niruri protein (PNP) possesses any protective role against aspirin mediated liver and spleen tissue toxicity, and if so, what signaling pathways it utilizes to convey its protective action. Aspirin administration in mice enhanced serum marker (ALP) levels, reactive oxygen species (ROS) generation, reduced antioxidant power and altered oxidative stress related biochemical parameters in liver and spleen tissues. Moreover, we observed that aspirin intoxication activated both the extrinsic and intrinsic apoptotic pathways, as well as down regulated NF-\u03baB activation and the phosphorylation of p38 and JNK MAPKs. Histological assessments and TUNEL assay also supported that aspirin induced tissue damages are apoptotic in nature. PNP treatment after aspirin exposure effectively neutralizes all these abnormalities via the activation of survival PI3k/Akt pathways. Combining all results suggest that PNP could be a potential protective agent to protect liver and spleen from the detrimental effects of aspirin.Aspirin has been used for a long time as an analgesic and anti-pyretic drug. Limitations of its use, however, remain for the gastro-intestinal side effects and erosions. Although the role of aspirin on gastro-intestinal injury has been extensively studied, the molecular mechanisms underlying aspirin Literature provides a consensus that conventional NSAIDs mediated ROS formation may suppress the risk of gastrointestinal related cancers Terminalia arjunaCajanus indicusPithecellobium dulcePhyllanthus niruriMany traditional ayurvedic herbs possess antioxidant properties. Examples are: Phyllanthus family are employed in ayurvedic formulation for the treatment of various diseases like urolithiasis P. niruri. It has been already reported that the aqueous extract P. niruri (PNP) possess the protective effects against various drugs and toxins mediated oxidative insults and pathophysiological complications Since earlier times, many species of c into the cytosol, caspase 3 as well as caspase 8 protein levels. Role of mitogen-activated protein kinase (MAPKs) and NF-\u03baB under this pathophysiological situation were also investigated in this study. The mode of cell death in ASA induced spleen and hepatotoxicity and the protective role of PNP has been investigated by histology, TUNEL assay and FACS analysis. The consequences of the present study are anticipated to draw a clear picture of the protective mechanism of PNP against ASA induced hepatic and spleen injury as well as it may also shed light on an achievable solution to the devastating complications of aspirin administration.The adverse effect of ASA administration and the protective action of PNP has been evaluated by measuring liver specific serum marker enzyme (ALP) leakage; lipid peroxidation, protein carbonylation; levels of cellular metabolites (GSH and GSSG) and activities of antioxidant enzymes . The molecular mechanism was determined by investigating the antiapoptotic Bcl-2 and pro-apoptotic Bax protein expressions, release of cytochrome 2SO4], 1-chloro-2,4-dinitrobenzene (CDNB), 5,5\u2032-dithiobis(2-nitrobenzoic acid) , ethylene diamine tetraacetic acid (EDTA), N-ethylmaleimide (NEM), nicotinamide adenine dinucleotide reduced (NADH), nitro blue tetrazolium (NBT), oxidized glutathione (GSSG), phenazine methosulphate (PMT), potassium dihydrogen phosphate (KH2PO4), reduced glutathione (GSH), sodium dihydrogen phosphate (NaH2PO4), sodium pyrophosphate, trichloro acetic acid (TCA), thiobarbituric acid (TBA), tris buffer, vitamin C were of the highest analytical grade and were bought from Sisco research laboratory . Bovine serum albumin (BSA) and Bradford reagent were purchased from Sigma-Aldrich Chemical Company, (St. Louis) USA. Antibodies such as anti Caspase-3 (ab47131), anti Caspase-8 (ab25901), anti Bid (ab77815), anti Bcl2 (ab7973), anti cytochrome c (ab76237), anti p38 (ab47363), anti JNK (ab76572), Phospho JNK (ab4821), anti Bax (ab32503), anti PI3k (ab74136), anti Akt (ab17785), Phospho Akt (ab23509), HRP (ab97051) were purchased from abcam . Anti NFkB (#3034), Phospho NFkB (#3031), Phospho p38 (#9211), anti PARP (46D11) was purchased from Cell Signaling Technology .Kits for ALT measurement were purchased from Span diagnostic Ltd., India. Ammonium sulphate [(NH4)ad libitum; exposed to 10\u201312 hours of daylight under standard conditions of temperature (25\u00b0C) and humidity (30%). All the studies with the experimental animals were performed following the standard ethical protocols of IAEC, Bose Institute, Kolkata. Full details of the study were approved by both IAEC and CPCSEA , Ministry of Environment and Forests, New Delhi, India (the permit number is: 95/99/CPCSEA).Healthy Swiss strain male albino mice weighing approximately 24\u201325 g were purchased from CNCRI, Kolkata, India. The animals were accustomed under laboratory conditions for a fortnight prior to experiments. They were maintained on a standard diet and water Phyllanthus niruri is a shrub belonging to the family Euphorbiaceae. Fresh young leaves were collected from Bose Institute experimental farm.The protein from Phyllanthus niruri (PNP) was isolated and purified following the method of Sarkar et al The homogeneity and the molecular weight of the protein was confirmed by SDS-PAGE with known molecular weight marker proteins (25\u2013225 kDa) by following the method of Sarkar et al. To check the biological activity of the purified PNP, the protein specific evidence-based experiments like effect of heat treatment and the effect of trypsin digestion have been carried out on PNP by following the techniques as described elsewhere To establish the dose of ASA necessary for hepatic damage, mice were randomly allocated into six groups each consisting of six animals and they were treated as follows: First group served as normal control (received only water as vehicle). Remaining five groups were treated with five different doses of ASA orally .Twenty-four hours after the final dose of ASA intoxication, all mice were sacrificed and ALP levels were measured using serum of all experimental mice.For the dose-dependent study, mice were randomly distributed into six groups each consisting of six animals. First two groups were served as normal control (received only water as vehicle) and toxin control respectively. Remaining four groups of animals were administrated with ASA followed by four different doses of PNP . Previously it was found that at this level of dose the P. niruri protein fraction protected liver against oxidative stress -dependent hepatic disorder, mice were divided into seven groups each consisting of six animals. First two groups were served as normal control (received only water as vehicle) and toxin control respectively. Other five groups of animals were treated with PNP intraperitoneally at a dose of 10 mg/kg body weight, once daily for 1, 1.5, 2, 2.5 and 3 weeks after ASA intoxication .To determine the time dependent effects of PNP treatment in ASAAt selected times after ASA and PNP treatment, all mice were sacrificed. ALP levels were measured using serum of all experimental mice.The animals were divided into five groups, consisted of six mice in each and they were treated as follows.\u201cNormal control\u201d: animals received only water as vehicle.\u201cToxin control (ASA)\u201d: animals received ASA at a dose of 100 mg/kg body weight once daily for 6 weeks, orally.\u201cPNP post-treated group (ASA+PNP)\u201d: animals were intraperitoneally injected with PNP for 2 weeks after ASA intoxication for 6 weeks.\u201cVitamin C post-treated group (ASA+VitC)\u201d: animals were intraperitoneally injected with Vitamin C for 2 weeks after ASA intoxication for 6 weeks.\u201cPNP alone treated group (PNP)\u201d: no treatment for first 6 weeks, later animals were treated with PNP for 2 weeks.The animals were sacrificed under light ether anesthesia and after that livers and spleens were collected.After scarification, the livers and spleens from experimental animals were quickly excised and weighed. Then the ratio of liver weight to body weight was measured for each.For assessment of serum specific marker (ALP levels) related to hepatic dysfunction, blood samples were collected by puncturing mice hearts of all experimental animals, kept overnight for clotting and then centrifuged at 3,000 g for 10 minutes. ALP level in the serum of experimented animals was measured by using standard kits according to the method of Kind and King Liver samples were homogenized using glass homogenizer in 100 mM potassium phosphate buffer containing 1 mM EDTA, pH 7.4 supplemented with protease and phosphatase inhibitors and centrifuged at 12,000 g for 30 minutes at 4\u00b0C. The supernatant was collected and used for the experiments.g for 30 minutes at 4\u00b0C. The supernatant was collected and used for the experiments.Spleen samples were also homogenized using glass homogenizer in 100 mM potassium phosphate buffer pH 7.4, containing 1 mM EDTA, 1 mM PMSF (proteinase inhibitor) and phosphatase inhibitor cocktail. The homogenized mixture was centrifuged at 12000\u00d7The protein content of experimental sets was measured following the method of Bradford (1976) using crystalline Bovine Serum Albumin (BSA) as standard.x) and glutathione-S-transferase (GST) have been measured in liver and spleen homogenates of all experimental animals.The activities of the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase formation and protein carbonylation were determined in liver and spleen homogenates of all experimental animals according to the method as described by Rashid et al. Reduced glutathione (GSH) and oxidized glutathione (GSSG) levels were estimated in liver and spleen homogenates of all experimental animals following the method of Ghosh et al. Ferric Reducing/Antioxidant Power (FRAP) assay was performed in order to determine the antioxidant power of liver as well as spleen tissues following the method of Benzie and Strain At first, the hepatocytes were isolated from the liver tissue of the experimental animals by following the method of Sarkar and Sil 2, 20 mM NaH2PO4, 30 mM glucose and 5 \u00b5M DCFDA) at 37\u00b0C for 15 minutes. The formation of DCF was measured at the excitation wavelength of 488 nm and emission wavelength of 610 nm for 10 minutes by using fluorescence spectrometer equipped with a FITC filter.The ROS production from spleen tissue and liver tissue homogenates were estimated separately by following the method of Rashid et al. c (1\u22361000 dilution), anti Bad (1\u22361000 dilution), anti Bax (1\u22361000 dilution), anti Bcl-2 (1\u22361000 dilution), anti p-38 (1\u22361000 dilution) and anti JNK (1\u22361000 dilution) at 4\u00b0C for overnight. The membranes were washed in TBST for 30 min and incubated with appropriate HRP conjugated secondary antibody (1\u22362000 dilution ) for 2 h at room temperature and developed by the HRP substrate 3,3\u2032-diaminobenzidine tetrahydrochloride (DAB) system .Proteins (50 \u00b5g) from each sample were separated by 10% SDS-PAGE and transferred to PVDF membranes. Membranes were blocked using BSA and incubated separately with primary antibodies such as anti- caspase3, anti- caspase8, anti PARP and anti NF-\u03baB (1\u22361000 dilution), anti Akt (1\u22361000 dilution), anti cytochrome Fresh mitochondria were isolated from the liver tissue Livers and spleens from the normal and experimental mice were fixed in 10% buffered formalin and were processed for paraffin sectioning. Sections of about 5 \u00b5m thickness were stained with haematoxylin and eosin to evaluate the histology under light microscope.The experimental paraffin embedded spleen tissue sections (5 \u00b5m) were warmed 30 minutes (64\u00b0C), deparaffinised and rehydrated. Terminal transferase mediated dUTP nick end-labelling of nuclei was performed by using APO-BrdU TUNEL Assay kit following the manufacturer\u2019s protocol.Normal and experimental hepatocytes were incubated with Annexin V and propidium iodide for 30 min at 37\u00b0C. Excess PI and Annexin V were washed off. After that, the cells were analyzed by flow cytometry using FACS Calibur equipped with 488 nm argon laser light source; 515 nm band pass filter for FITC-fluorescence and 623 nm band pass filter for PI-fluorescence using Cell Quest software.All experimental values have been represented as mean \u00b1 S.D. (n\u200a=\u200a6). Data on biochemical investigation were analyzed using analysis of variance (ANOVA) and the group means were compared by Duncan\u2019s Multiple Range Test (DMRT). P values of 0.05 or less were considered significant.At the beginning of our present study we have purified PNP and confirmed its biological activities via heat treatment and enzymatic digestion analysis. Later we evaluated its antioxidant activities against drug induced pathophysiology Aspirin (ASA) is safe and broadly utilized NSAID when used at the therapeutic level but higher doses or prolonged use of this drug may promote oxidative stress and result in gastrointestinal erosions and apoptotic lesions It is reported that aspirin causes hepatotoxicity and elevates the levels of serum marker enzymes Survival of cells under oxidative stress is an important parameter to evaluate the effectiveness of any prophylactic agent. The result of our study suggests that ASA at a dose of 100 mg/kg body weight up regulated the ALP level in plasma but that could be reversed with the treatment of PNP up to a dose of 10 mg/kg body weight daily up to 2 weeks . Time deAfter the fixation of the dose and treatment time, we designed our animal experimental protocol and perfThe result of present study showed that ASA administration for 6 weeks reduced the liver weight to body weight ratio . TreatmeIn mammalian tissue the complex endogenous antioxidant system and chemical sequesters help to prevent oxidative damage. In our study we explored the effect of the drug (ASA) on the activities of antioxidant enzymes in both the liver and spleen tissues. The results showed that ASA administration at the hepatotoxic dose reduced the activities of antioxidant enzymes in both hepatic and spleAs expected, ASA at hepatotoxic dose caused a significant reduction in FRAP value but postThe protein carbonyl group is generated by ROS through many different mechanisms and its concentration is a good measure of protein oxidation via oxidative stress. The side chains of all amino acid residues of proteins are susceptible to oxidation by the action of ROS. Literature suggests that enhanced protein carbonylation may be responsible for the decrease in antioxidant enzyme activity Reduced glutathione is present at high concentrations in all mammalian cells, especially in the renal cells, hepatocytes, and erythrocytes. GSH is the main non-protein thiol intracellular antioxidant that scavenges free radicals. A considerable amount of GSH is consumed to scavenge ROS 2O2 content thereby enhanced ROS formation in the liver. Treatment with PNP reversed the phenomenon of the ASA administrated fluorescence intensity of DCF signal in the liver and anti-apoptotic (Bcl-2) mitochondrial membrane proteins as well as the activation of caspase cascades n of Bid . TRAIL bn of Bid and TUNEn of Bid . Besidesn of Bid and splen of Bid could beAnother essential serine/threonine protein kinase that plays a vital role in cell survival pathways by inhibiting apoptosis is Akt. Activation of Akt requires the activation of PI3K and might promote cellular survivality Overall, the present study demonstrated that like other NSAIDs, ASA administration at hepatotoxic dose induces ROS formation. Once formed, ROS altered the normal GSH/GSSG balance and at the same time caused lipid peroxidation of the cellular membrane. Most importantly, the cellular endogenous antioxidant defense mechanisms are also disrupted and thereby shifting the physiological redox status. Besides, ROS mediated activation of JNKs and p38 MAPKs altered the balance between the pro-apoptotic and anti-apoptotic proteins. Under these circumstances, the overall system might follow both intrinsic and extrinsic apoptotic signaling pathways. However, PNP treatment after ASA intoxication might be able to protect the liver and spleen via 1) scavenging free radicals thereby inhibiting ROS formation, 2) enhancing the antioxidant enzyme activities and maintaining the proper GSH/GSSG ratio, 3) ameliorating ASA mediated inhibition of NF-\u03baB and thereby activation of anti-apoptotic Bcl-2 proteins as well as inhibition of MAPKs activation and 4) by activating Akt/PI3k mediated cell survival signaling pathways. A possible mechanism of PNP induced survival signaling pathways against ASA induced hepatic and spleen toxicity has been depicted in Figure S1Schematic representation of group division for mice treatment in the study.(TIF)Click here for additional data file."} +{"text": "Tight glucose control (TGC) failed to improve survival and has been associated with high rates (6-18%) of hypoglycemia (< 2.5mmol/l), themselves associated with significant mortality. Blood glucose levels vary largely in critically ills and TCG requires individualized knowledge of the patient condition. Bedside frequent blood glucose measurements are mandatory for continue adaptations of insulin and glucose administration. We progressively transfer TGC to the nursing personal.TGC was introduced in 2003 and then progressively transferred from physicians to nurses since 2007. Nurses are specifically trained to adapt infusion rates of nutrition and insulin according to medically predefined targets . Glucose levels are determined by the central lab. or by ICU blood gas analyzers. Glycemia (n=809\u2019725) were extracted from our electronic clinical information system (Metavision\u00ae) and analyzed with STATA.A continuous decrease in yearly standard deviations (IQR), suggest that TGC was effectively improved (Table).We suppressed the lowest target (4.5\u20136.0 mmol/l) in May 2009 and this may explain the mean increase since then. The rate of hyperglycemia decreases below 10%, with rates of hypoglycemia (<2.5 mmol/l) largely below 0.4%, 50 to 100 fold lower than those reported in the literature.Implementation and transfer of tight glucose control in a large mixed adult ICU significantly decreases the rate of hyperglycemia to less than 10%, with extremely low rates of hypoglycemia (< 0.1%). These results strongly suggest that transfer of tight glucose control to bedside nursing personal is safe and effective.None declared."} +{"text": "Podocyte-specific Scribble knockout mice develop normally and display no histological, ultrastructural or clinical abnormalities up to 12 months of age. In addition, no increased susceptibility to glomerular stress could be detected in these mice. In contrast, constitutive Scribble knockout animals die during embryonic development indicating the fundamental importance of Scribble for embryogenesis. Like in podocyte-specific Scribble knockout mice, the development of podocyte foot processes and the slit diaphragm was unaffected in kidney cultures from constitutive Scribble knockout animals. In summary these results indicate that basolateral polarity signaling via Scribble is dispensable for podocyte function, highlighting the unique feature of podocyte development with its significant apical membrane expansions being dominated by apical polarity complexes rather than by basolateral polarity signaling.The kidney filter represents a unique assembly of podocyte epithelial cells that tightly enwrap the glomerular capillaries with their complex foot process network. While deficiency of the polarity proteins Crumbs and aPKC result in impaired podocyte foot process architecture, the function of basolateral polarity proteins for podocyte differentiation and maintenance remained unclear. Here we report, that Scribble is expressed in developing podocytes, where it translocates from the lateral aspects of immature podocytes to the basal cell membrane and foot processes of mature podocytes. Immunogold electron microscopy reveals membrane associated localisation of Scribble predominantly at the basolateral site of foot processes. To further study the role of Scribble for podocyte differentiation The glomerular filtration barrier is a unique structure characterized by a precise three dimensional framework of podocytes that elaborate long, regularly spaced, interdigitating foot processes, enveloping the glomerular capillaries. Neighbouring podocytes are connected by the slit diaphragm, a specialized cell junction and the only cell-cell contact of mature podocytes, that bridges the filtration slit between podocyte foot processes In epithelial cells apicobasal cell polarity is established by the asymmetric distribution of three core polarity complexes, the apical Crumbs complex, consisting of Crumbs, PALS1 and PATJ, the apical Par complex localizing at the tight junctions and the basolateral Scribble complex, comprising the proteins Scribble, Dlg and Lgl Scribble is a large cytoplasmic scaffold protein of the leucine-rich repeat (LRR) and PDZ domain (LAP) family with 16 N-terminal LRRs and 4 C-terminal PDZ domains Scribble (circletail and rumpelstilzchen mutations) cause severe impairment of neural tube development with craniorachischisis and neonatal death loop-tail mouse In mice, point mutations of Scribble heterozygosity causes prostate hyperplasia, while prostate-specific knockout of Scribble results in loss of cellular polarity, elevated proliferation and progression to intraepithelial neoplasia Scribble is targeted to proteasomal degradation by the high risk papilloma virus protein E6-E6AP ubiquitin-protein ligase complex Scribble knockout mice to investigate the role of Scribble in podocyte differentiation and maintenance.Here we analysed the spatiotemporal expression of Scribble during glomerular development and generated podocyte-specific and constitutive Previously, we identified that the aPKC complex translocates from the apical to basal membranes during podocyte differentiation, preceding the development of primary and foot processes flox/floxScribble mice were generated by introducing loxP-sites into the introns 1 and 8 flanking Scribble exons 2\u20138 (flox/floxScribble mice were crossed with NPHS2.Cre mice to create podocyte-specific Scribble knockout mice (\u0394podocyteScribble) (To study the role of Scribble for podocyte differentiation xons 2\u20138 A. Scribodocyte) B. Whileodocyte) C, D.\u0394podocyteScribble mice displayed a regular glomerular architecture (\u0394podocyteScribble mice (\u0394podocyteScribble and control mice (\u0394podocyteScribble mice featured no increased susceptibility to glomerular stress in the BSA overload and the Adriamycin (ADR) model model G, H.Cre expression under control of the NPHS2-promoter starts late during glomerular development Scribble knockout in podocytes might be masked in flox/flox; NPHS2.CreScribble mice. To study the effect of Scribble knockout on early glomerular development and podocyte differentiation, we analyzed kidneys of constitutive Scribble knockout mice. flox/floxScribble mice were crossed with Cre deleter mice to create constitutive Scribble knockout mice were further backcrossed to C57BL/6J mice. Cre-mediated recombination causes a frame shift and early stop of translation. In addition floxed mice were crossed to C57BL/6J Cre deleter mice to excise the loxP flanked genomic region (exons 2\u20138) and to generate heterozygous mice carrying the constitutive knockout allele (\u2212/+Scribble) flox/+Scribble and \u2212/+Scribble mice were performed by genOway .LoxP sites were introduced into the introns 1 and 8 flanking NPHS2.Cre mice were kindly provided by Lawrence Holzman Scribble-floxed mice (flox/floxScribble) were crossed with NPHS2.Cre mice to generate podocyte-specific Scribble knockout mice flox/flox; NPHS2.CreScribble (\u0394podocyteScribble). NPHS2.Cre negative flox/+Scribble and flox/floxScribble litter mates served as controls. Mice carrying the circletail allele of Scribble were purchased from The Jackson Laboratory.Scribble exons 2\u20138 DNA was isolated from tail clip or isolated glomeruli, respectively. For detection of WT or the loxP site, the primers were tccagttagcactcaggcgtcagg (forward) and cagctccgagaggttctcacagtcc (reverse). For detection of the deletion, primers were accccagtgctctctggtgtttttattg (forward) and cagctccgagaggttctcacagtcc (reverse). For detecting the circletail mutation, primers were ctagccctcccccccc and cctgggactgagaaggacat (reverse). All animal studies were approved by the Committee on Research Animal Care, Regierungspr\u00e4sidium Freiburg and by the Washington University Animal Studies Committee.For genotyping or detection of deletion of \u0394podocyteScribble mice and control littermates (n\u200a=\u200a5 each) received endotoxin-free BSA (Sigma A9430) intraperitoneally for 4 consecutive days (10 mg/g body weight) \u0394podocyteScribble mice and control littermates (n\u200a=\u200a6 each) received one dose of intravenous Adriamycin (Sigma) in 0.9% NaCl (2 mg/ml) (15 \u00b5g/g body weight) Urinary albumin and urinary creatinine were measured using a fluorimetric albumin test kit (Progen) and an enzymatic creatinine kit (Labor+Technik) following the manufacturer's instructions. Proteinuria was expressed as mg albumin/mg creatinine.Scribble heterozygous knockout mice; the date of the vaginal plug was designated as day 0. Metanephric kidneys were microdissected from the embryos at embryonic day 12.5 and cultured in Dulbecco's modified Eagle's medium containing 10% fetal calf serum and 1% Penicillin and Streptomycin at 37\u00b0C and 5% CO2 on 0.4 \u00b5m transwell inserts Timed matings were set up with constitutive Kidneys were fixed in 4% paraformaldehyde, embedded in paraffin or Epon and further processed for PAS staining or transmission electron microscopy, respectively.Kidneys were frozen in OCT compound and sectioned at 6 \u00b5m (Leica Kryostat). The sections were fixed with 4% paraformaldehyde, blocked in PBS containing 5% BSA and incubated for 1 hour with primary antibodies as indicated. After PBS rinse for several times, fluorophore-conjugated secondary antibodies (Invitrogen) were applied for 30 minutes. Images were taken using a Zeiss laser scan microscope equipped with a 63\u00d7 water immersion objective or a Zeiss fluorescence microscope equipped with a 5\u00d7, a 20\u00d7 and a 40\u00d7 oil immersion objective. To determine the number of podocytes per glomerular section, kidney sections were stained against the podocyte nuclear marker WT1. WT1-positive cells were counted in 30 glomeruli per mouse per condition (n\u200a=\u200a3 for each condition).Fixed samples of rat kidney were embedded in Lowicryl K4M resin (Electron Microscopy Sciences), and ultrathin sections were labeled by an indirect immunogold protocol, as described Antibodies were obtained from Millipore , Progen , Sigma , Abcam and Santa Cruz Biotechnology . Mouse monoclonal antibody against rat Podocalyxin was described before Data were expressed as the mean \u00b1 SEM. Statistical comparisons were performed using two-tailed Student's t-test if not stated otherwise. Differences with P<0.05 were considered significant.Figure S1Expression of Scribble in the developing and adult kidney. Frozen kidney sections of newborn and adult Wistar rats were stained using antibodies against Scribble and the podocyte marker proteins (A) WT1 and (B) Nephrin. Scribble is expressed in glomeruli as well as in segments of the tubule system. Scale bars: 50 \u00b5m.(TIF)Click here for additional data file.Figure S2Expression pattern of Par3 in glomeruli of \u0394podocyteScribble mice. Frozen kidney sections of adult \u0394podocyteScribble and control mice were stained using antibodies against Par3 and the slit diaphragm protein Nephrin. No difference in the expression pattern of Par3 could be detected in \u0394podocyteScribble compared to control mice. Arrows indicate colocalisation of Par3 and Nephrin. Scale bars: 5 \u00b5m.(TIF)Click here for additional data file."} +{"text": "HIV heterosexual transmission mainly occurs by exposition of female genital mucosa with infected male seminal secretions. The notion of compartmentalization of HIV in the male genital tract is well established by the demonstration of the existence of several viral populations between blood and semen, especially in the gp160 viral envelope. The objectives of the present work were (i) the study of the role of genital epithelial cells in the heterosexual transmission of HIV and (ii) the in vitro modeling of this crossing by using chimeric viruses (pBrNL4.3-eGFP/dsRedExpress) that express the gp160 glycoprotein isolates from patients\u2019semen.Seminal samples that showed a positive viral load were selected from 50 semen specimens obtained from 39 HIV-infected patients followed at the University-Hospital ofSaint-Etienne. Chimeric viruses were constructed by cloning a pBrNL4.3 vector with a viral envelope containing a GFP or a dsRedExpress fluorochrome. HEC genital epithelial cells were infected by CXCR4-tropic (LAI) or CCR5-tropic (BaL) HIV-1 viruses (0.001 MOI per cell during 24h) in different conditions (presence or absence of AZT or proinflammatory cytokines).Two days after infection, intracellular viral RNA was detected for both virus strains (LAI and BaL) either in the presence or in the absence of AZT. By contrast, the detection of proviral DNA (LTR) was observed only for the CXCR4-tropic variant (500 \u00b1 100 copies / 106 cells); the addition of AZT abolished this infection. Neither intracellular viral proteins nor extracellular viral RNA could be detected in infected cells.These results confirm the selective sequestration of CXCR4-tropic viruses by epithelial cells. Complementary analyses are ongoing for determining the exact role of these cells in HIV-1 heterosexual transmission and as a viral latency reservoir that could be reactivated under inflammatory conditions. A confocal microscopy imaging using the chimeric viruses will complete these preliminary results."} +{"text": "We present our patients we treated with carotico-subclavian bypass operation for subclavian steal syndrome or as an adjunctive procedure to thoracic endovascular stent- grafting (TEVAR) procedures necessiating left subclavian artery occlusion intentionally.We performed 11 carotico-subclavian bypass operations between August 2009-January 2013 at our department. 8 patients were operated for subclavian steal syndrome with subclavian artery occlusion and 3 patients were operated before TEVAR for aortic dissection/aneursym necessiating left subclavian artery coverage. 9 male (81.81%) and 2 female (18.18%) patient with the age between 55-73 (mean 64.9) were operated. 8mm Dacron graft was used for nine patients and 7/8mm PTFE for two patients. Anastomosis were done in end-to-side fashion.Radial artery pulses were palpabl in all patients after the operation with resolving complaints as left arm pain or dizziness.1patient with native carotis artery tortiosity and cerebral artery aneursym suffered graft thrombosis 2 days after the operation (he could not be anticoagulated). He was reoperated and graft was cut near carotis anastomosis. After removing thrombotic material and providing flow inside graft, reanastomosis to a more distal and straight part was performed. No neurological complications were observed. 1 patient suffered puffiness and pain at the left shoulder two weeks after operation because of seroma that was drained percutaneously with ultrasonography guidance. Patients to have TEVAR procedures were primarily operated for carotico-subclavian graft interposition and subsequently TEVAR was performed at the same stage. The mean follow-up period is 20 months with all grafts having patency on ultrasonographic examination.Carotico-subclavian bypass is a safe procedure with good surgical results."} +{"text": "Recent data suggest an important role of zidovudine (ZDV) and interferon-\u03b1 (IFN-\u03b1) in improving response rates and survival in acute ATLL. Treatment of chronic ATLL with ZDV/IFN-\u03b1 alone has recently been associated with 100% survival beyond five years.Retrospective analysis of patients with acute and chronic ATLL treated with ZDV/IFN-\u03b1 first line. Response was assessed one month from the start of treatment using total lymphocyte and CD4 count, HTLV-1 proviral load (PVL) and clonal analysis (in house method).Acute ATLL: response rate 33% . Median overall survival (OS) 3 months (range 3-8).Chronic ATLL: response rate 100% . Median OS 20 months (range 9-73). In chronic ATLL these prolonged responses were observed despite lower dose therapy. Two patients, showed 10-fold reductions in PVL which occurred more than 1 year after haematological CR. All patients remain in remission at time of analysis. Clonality studies demonstrated a dominant clone at base line with emergence of a polyclonal pattern after viral load reduction.The complete response in one patient with acute ATLL supports the recent observation that ZDV/IFN-\u03b1 is effective as first line treatment in some patients.The significant reduction in PVL and late emergence of a polyclonal integration pattern suggest benefit from prolonged ZDV/IFN-a therapy in chronic ATLL and the utility of both PVL and clonal analysis as a test of the efficacy of novel treatment regimes."} +{"text": "Ankyrin repeat domain 12 (ANKRD12), is encoding a 224\u00a0kDa nuclear protein and most conserved at its N-terminal ankyrin repeats region and the C-terminal co-activator interacting domain. The aim of this study was to investigate the ANKRD12 mRNA expression in colorectal cancer (CRC) tumor tissues and the normal adjacent mucosa and its potential relevance to clinicopathological characteristics and prognosis.Surgical specimens of tumor tissues (n\u2009=\u200968) and adjacent normal mucosa (n\u2009=\u200951) were obtained from CRC patients. The ANKRD12 mRNA expression was measured by quantitative real time reverse transcriptase polymerase chain reaction. The relationship between ANKRD12 mRNA expression and clinicopathological features was analyzed by appropriate statistics. Kaplan\u2013Meier analysis and Cox proportional hazards regression models were used to investigate the correlation between ANKRD12 expression and prognosis of CRC patients.The relative mRNA expression of ANKRD12 were significantly lower in CRC tumor tissues than in the normal adjacent mucosa (P\u2009<\u20090.001), and the cases with low ANKRD12 expression showed a higher frequency of liver metastasis (P\u2009=\u20090.015). Kaplan\u2013Meier analysis indicated that patients (CRC without liver metastasis) with low ANKRD12 expression had poor overall survival (P\u2009=\u20090.041). Multivariate analysis showed that low ANKRD12 expression was an independent predictor of overall survival.This study revealed that ANKRD12 mRNA were down regulated in CRC tumor tissues and low ANKRD12 expression was correlated with liver metastasis and poor survival of CRC patients. Colorectal cancer (CRC) is one of the leading causes of cancer mortality in the world. An estimated 143,460 new cases of CRC occurred in 2012 and 51,690 people have died from CRC during the same year to investigate ANKRD12 mRNA expression in association with clinicopathologic variables , the overall survival rate was significantly lower in the patients with low ANKRD12 mRNA expression than that in those with high expression is crucial in many physiological processes. The activity of NRs is first regulated by ligands , as bindAnkyrin repeat domain 11 (ANKRD11), also called ANCO-1, is located within the 16q24.3 breast cancer loss of heterozygosity (LOH) region and was The precise molecular mechanisms behind the altered expression of ANKRD12 in colorectal cancer are unclear. To our knowledge, this is the first report to describe the significance of ANKRD12 to clinical stage, lymph node and liver metastases, and prognosis of CRC patients. ANKRD12 binds to alteration/deficiency in activation 3(ADA3) through its C-terminal domain and inhibits ADA3-mediated transcriptional co-activation on NRs . ADA3 isIn conclusion, we found that ANKRD12 mRNA were downregulated in CRC tumor tissues and low ANKRD12 mRNA expression correlated with poor overall survival and liver metastasis of CRC patients. These findings suggest that ANKRD12 is a cancer-related gene associated with liver metastasis and a survival predictor of CRC patients.Written informed consent was obtained from the patient for publication of this report and any accompanying images.ANKRD12: Ankyrin repeat domain 12; CRC: Colorectal cancer; ANCO: Ankyrin repeats-containing cofactor; HDACs: Histone deacetylases; ANKRD11: Ankyrin repeat domain; LOH: Loss of heterozygosity; RAC3: Ras-related C3 botulinum toxin substrate 3; qRT-PCR: Quantitative real time reverse transcriptase polymerase chain reaction; NRs: Steroid/nuclear hormone receptors; ADA3: Alteration/deficiency in activation 3.The authors declare that they have no competing interests.RB, DL and SZ conceived and designed the study, performed the experiments and wrote the paper. ZS and XFF contributed to the writing and to the critical reading of the paper. WTG performed patient collection and clinical data interpretation. All authors read and approved the final manuscript."} +{"text": "The benefit of screening asymptomatic diabetic patients for CAD remains unclear. A recent ADA consensus statement on this issue concluded that routine screening is not recommended. The aim of this study was to assess the yield of stress perfusion CMR in a diabetic population.All patients who underwent stress CMR at ICPS between November 2009 and September 2010 were identified through the use of the cardiology database. Exclusion criteria included (1) history of documented myocardial infarction (n = 623); (2) prior percutaneous coronary intervention (PCI) (n = 1335); and (3) prior coronary artery bypass grafting (CABG) (n = 264). For patients who underwent multiple tests during this time period (n = 116), only the first test was included. The study population consisted of 2737 patients.P =0.48). However diabetic patients were more likely to have inducible ischemia than nondiabetic patients , whereas they were equally likely to have evidence of infarction on delayed enhancement images (23.7% vs 25.4%).Similar percentages of diabetic and nondiabetic patients had abnormal scans , and infarction .With regards to symptomatic patients, patients with diabetes had significantly higher percentages of ischemia , but similar amounts of infarction than symptomatic nondiabetic patients.Ejection fractions after stress testing were similar in both groups (diabetic vs nondiabetic - 68.47% vs 69.11% p = ns).Stress testing in asymptomatic diabetics identifies a high percentage of patients with unknown myocardial infarction. This information has important prognostic implications. Asymptomatic diabetic patients have a significantly higher amount of ischemia than non-diabetic patients. Ischemia is the main predictor of adverse cardiac outcomes. Early identification of this can guide revascularization, which has been shown to improve prognosis in those with at least moderate ischemia."} +{"text": "Multiple Myeloma (MM) is a malignant proliferation of B-cells with plasma cell differentiation. Interactions of plasma cells with stromal cells and other cells in bone marrow (BM) microenvironment are responsible for MM progression. Screening of proteins expressed on the membrane surface of plasma cells, stromal cells and mononuclear cells from MM patients by mass spectrometry (MS) may identify new prognostic and therapeutic targets in this incurable disease.in vitro expanded and characterized by immunocytochemistry and flow cytometry . Proteins were obtained by Pierce Cell Surface Protein Isolation kit and quantified by Pierce 660nm Protein Assay.BM from eight MM patients, five normal BM from donors and seven tonsils of children submitted to tonsillectomy (controls) were collected. Plasma cells (CD138+) from MM patients and tonsils, stromal cells (CD105+) and mononuclear cells from MM patients and normal BM were isolated by magnetic cell sorting (MACS). Stromal cells were In vitro expansion of stromal cells was possible for four MM and four normal BM samples. Stromal cells were CD105+, Vimentin+, CD45- and CK-, without contamination with other cell types. Plasma cell pool from MM patients or tonsils (control), stromal cell pool and mononuclear cell pool from MM patients or normal BM (control) were subjected to protein extraction (yield ranged from 50\u03bcg to 100\u03bcg). Analysis of proteins will be performed by LTQ (Linear Ion Trap Quadropole) Orbitrap Velos Mass Spectrometry and the spectra MS/MS will be obtained using Mascot.S\u00e3o Paulo Research Foundation (FAPESP), Brazil."} +{"text": "Previous in vivo studies on total ankle arthroplasty (TAA) kinematics were mainly performed using skin marker analysis, which has the drawback of skin movement artefacts . A furthThe kinematics of 11 TAA participants were simultaneously analysed by skin marker and videofluoroscopic assessment during level gait (gt), walking up- (uph) and downhill (dnh). The fluoroscopic data analysis included a 2D/3D registration [Skin marker analysis significantly overestimated sagittal plane ROM of the TAA for 5(gt), 6(uph) and 6(dnh) and underestimated for 1(uph) and 2(dnh) subjects. Frontal plane ROM was significantly overestimated for 7(gt), 8(uph) and 9(dnh) of the 11 subjects. Transverse plane ROM was for 2(uph) and 2(dnh) subjects significantly overestimated, and for 3(gt), 1(uph) and 7(dnh) subjects significantly underestimated by skin markers. For mean RMS diff, mean max diff and mean ROM see Table The differences between skin marker assessed rearfoot-shank and the fluoroscopic assessed isolated TAA motion were neither consistent between subjects, nor motion planes, nor conditions. For transverse and frontal plane rotations, the maximal differences were in the range of the maximal corresponding ROM. Discrepancies for the sagittal plane were smaller, but still for some subjects, ROM were significantly different."} +{"text": "The aim of the present work was to characterize the electrophysiological effects of the non-steroidal anti-inflammatory drug diclofenac and to study the possible proarrhythmic potency of the drug in ventricular muscle.Ion currents were recorded using the voltage clamp technique in canine ventricular cells, and action potentials (AP) were recorded from canine ventricular preparations using microelectrodes. The proarrhythmic potency of diclofenac was investigated in an anaesthetized rabbit proarrhythmia model.Kr) and slow (IKs) delayed rectifier and L-type calcium currents (ICa) without influencing transient outward (Ito) and inward rectifier (IK1) potassium currents. The action potential was slightly lengthened in ventricular muscle but shortened in Purkinje fibres by diclofenac (20 \u00b5M). The maximum upstroke velocity (Vmax) was decreased in both preparations. Larger repolarization lengthening was observed when repolarization reserve was impaired by previous BaCl2 application. Diclofenac (3 mg/kg) did not prolong the QTc interval, while the potassium channel blocker dofetilide (25 \u00b5g/kg) significantly lengthened QTc in anaesthetized rabbits. The combination of diclofenac and dofetilide significantly prolonged QTc. Diclofenac alone did not induce torsades de pointes ventricular tachycardia (TdP) while TdP incidence following dofetilide was 20%. However, the combination of diclofenac and dofetilide led to a significant increase in the incidence of TdP.Diclofenac (30 \u00b5M) decreased the amplitude of rapid (IThe results indicate that diclofenac, at therapeutic concentration and even at high dose, does not increase the risk of arrhythmia in normal heart. However, high dose drug treatment may enhance the proarrhythmic risk in the heart when the repolarization reserve is reduced."} +{"text": "Patients with bicuspid aortic valve (BAV) are at increased risk for both valvular and vascular complications. Preoperative knowledge about AV morphology, the presence and extent of AV and annulus calcification, valvular complications, and ascending aorta (AA) diameter is very important for surgical planning in patients with BAV.To evaluate the value of cardiac magnetic resonance imaging (CMR) for the comprehensive assessment BAV in comparison with transthoracic echocardiography (TTE), dual-source computed tomography (DSCT), and operative findings.Seventy-three consecutive patients with BAV who underwent TTE, DSCT, cardiac MRI and valve surgery were enrolled in this study. All CMR studies were performed with a 1.5 T whole-body system using an 8-element phased array surface coil. Two independent radiologists assessed the type of BAV (classified according to number of raphe), functional status of the BAV, AA diameter and left ventricular ejection fraction (LVEF). Stenotic aortic valve area (AVA) was computed with the continuity equation on TTE and by direct planimetry on DSCT and CMR. We compared quantitative grading of aortic regurgitation (AR) by CMRI with semiquantitative grading of AR by TTE. AA diameter was measured at 4 levels from mid-systole on DSCT and CMR. LVEF by CMR (Simpson\u2019s method) was compared with the use of TTE (M-mode method).\u03ba=0.954) of the underlying BAV complications as compared with TTE . Stenotic AVA (n=51) by CMR (0.88\u00b10.29 cm2) correlated well with AVA by DSCT and TTE . Regurgitation grading by CMR was significantly correlated with the grading of AR severity by TTE . AA dilatation more than 4.5 cm in diameter was present in 30 (41%) patients. There was excellent correlation in the mean diameter of AA between DSCT and CMR . A moderate correlation between CMR and TTE was shown for the evaluation of LVEF .Patients underwent AV repair (n= 72) or replacement (n=1) with AA graft replacement (n=3) or wrapping (n=28). CMR showed 91.8% agreement of BAV type compared with operative findings [no raphe (n=30) and raphe (n=43)]. CMR showed excellent agreement (CMR allows accurate imaging technique for comprehensive assessment of patients with BAV."} +{"text": "To the Editor: Adult vaccination rates are low to provide workplace vaccination, but we found little or no information about these organizations in the literature. Therefore, we interviewed community vaccinators about their 2009 experience with workplace vaccination against seasonal influenza virus and pandemic (H1N1) 2009 virus, their business practices, barriers encountered, and delivery of other adult vaccines.We selected a diverse study population of community vaccinators. We combined the 10 US Department of Health and Human Services regions to create 5 study regions. Beginning with a list of vaccinators provided by the Centers for Disease Control and Prevention and searching with Google for \u201con-site vaccinators,\u201d we identified 17 national and 28 local vaccinators (full list available from the authors). We selected at least 1 national and 1 local vaccinator from each region and then purposively sampled them to increase geographic and organizational diversity. Our sample comprised 5 national vaccinators, 7 local vaccinators serving urban and rural workplaces, a mobile-clinic vaccinator, a visiting nurses association, and an occupational health specialist.The qualitative study and a mismatch between vaccine demand and supply, resulting in delayed or lost business (9/12 respondents). Some vaccinators found the season more challenging than in prior seasons , yet most reported having added clients .Vaccinators\u2019 reported business practices include vaccinating at sites in addition to workplaces, for example, churches and faith-based settings (9 vaccinators), schools (9), and community centers (8). Most (9) reported vaccinating on multiple work shifts and at multiple worksites. Ten vaccinators also reported they can help employers publicize workplace vaccination events. Most did not report patient-level vaccination information to health plans (10 vaccinators), primary-care providers (9), or registries (8). Many directly bill Medicare (8) and private insurers (7) if asked.Additional findings related to barriers and delivery of other vaccines. Commonly reported barriers to increasing workplace vaccination rates were worker reluctance (10 vaccinators); worker out-of-pocket costs (9); and low worker awareness of workplace vaccination events (5). Other vaccines offered by these workplace vaccinators included the following: tetanus-diphtheria-pertussis (10 vaccinators), pneumococcal (10), hepatitis A and B (7), and herpes zoster (4).,This qualitative study, although small and not necessarily representative, found remarkable consistency across community vaccinators. Vaccinators were challenged by the pandemic (H1N1) 2009 vaccination season, but the season also provided new clients. Most reported vaccinating at diverse sites in addition to workplaces, and most already vaccinated against diseases other than influenza. Vaccinators consistently identified workers\u2019 reluctance and out-of-pocket costs, and poor publicizing of workplace vaccination events as remediable barriers to vaccination. Tackling of these barriers is supported by the literature ("} +{"text": "After the initial formation of a highly branched vascular plexus, blood vessel pruning generates a hierarchically structured network with improved flow characteristics. We report here on the cellular events that occur during the pruning of a defined blood vessel in the eye of developing zebrafish embryos. Time-lapse imaging reveals that the connection of a new blood vessel sprout with a previously perfused multicellular endothelial tube leads to the formation of a branched, Y-shaped structure. Subsequently, endothelial cells in parts of the previously perfused branch rearrange from a multicellular into a unicellular tube, followed by blood vessel detachment. This process is accompanied by endothelial cell death. Finally, we show that differences in blood flow between neighboring vessels are important for the completion of the pruning process. Our data suggest that flow induced changes in tubular architecture ensure proper blood vessel pruning. The vasculature is the first organ system to form during embryonic development and meets the challenge to grow and refine while it is already functioning. After the initial sprouting of blood vessels, remodeling ensures the formation of a more efficient vascular network Here, we use time-lapse imaging to analyze the cellular mechanisms that take place during the pruning of a defined blood vessel in the eye of zebrafish embryos. Our analysis reveals that angiogenic sprouting initially leads to the formation of a Y-shaped blood vessel branch, which is subsequently resolved. This process entails the rearrangement of endothelial cells within the pruning blood vessel from a multicellular to a partially unicellular tube. We show that blood flow is an important regulator of the pruning event. Importantly, our results suggest that loss of perfusion in itself does not lead to blood vessel pruning, but that pruning might be facilitated by the establishment of differences in blood flow between vessels in a branch point.sa916Tg(kdrl:Hsa.HsRAS-mcherry); y7Tg(fli1a:nEGFP) fish to label endothelial cell membranes and nuclei and 48 hpf in d nuclei . These md nuclei . Initiald nuclei . After bmu125Tg(kdrl:cytobow1.0) fish to label individual endothelial cells in different colors. Initially, endothelial cells of the dorsal CrDI formed a multicellular tube, with two cells surrounding the vascular lumen fish To corroborate our findings, we used Previous studies had shown that macrophage induced apoptosis was essential for the regression of hyaloid vessels during mouse development bcl2bcl2 injected embryos more endothelial cells migrated into the neighboring blood vessels. We therefore conclude that even though endothelial cell apoptosis occurs during CrDI pruning, it is not absolutely required for the completion of this process.To address the question of whether the observed apoptosis was required for blood vessel regression, we overexpressed the apoptosis inhibitor pu.1 MO injected embryos had normal regression in 95.1% of 352 eyes analyzed, compared to 97.8% in control embryos targeting the macrophage specific transcription factor pu1.1 Previous studies analyzing blood vessel pruning in zebrafish brains, mice retinae and airways showed that loss of blood vessel perfusion precedes blood vessel regression Based on these observations, we set out to determine whether manipulating blood flow patterns would influence CrDI pruning. We asked, whether we could establish forced blood flow through the dorsal CrDI by laser ablating the NCA. We performed the laser ablation on one side of experimental embryos, while we left the other side as a control. We observed a persistent dorsal CrDI, which continued to carry blood flow (data not shown), on the side on which we had ablated the NCA, but not on the control side . These rcardiac troponin t2a (tnnt2a), which never establish a heartbeat tnnt2a MO injected embryos, the CrDI failed to prune in about 50% of cases (Table S4 in File Supplementary Tables). In embryos that were either treated with 4x Tricaine or with Nifedipine to stop the heartbeat We next asked whether inhibition of blood flow would lead to precocious pruning of the CrDI. We manipulated blood flow by altering the heartbeat either by drug treatment or by injecting zebrafish embryos with MO targeting Our study is the first to describe the cellular rearrangements occurring during blood vessel pruning and establishes a sequence of events during which a multicellular tube is transformed into a partially unicellular tube prior to blood vessel detachment. This change in blood vessel architecture might be necessary in order to ensure proper blood vessel pruning without vessel rupture or hemorrhage. A recent report describes that during blood vessel fusion, endothelial cells rearrange from a unicellular to a multicellular tube per se that determines if a given blood vessel will prune. Rather, the difference in blood flow between neighboring blood vessels might be sensed by endothelial cells, ultimately triggering the pruning of the less perfused vessel. Our blood flow blocking experiments furthermore suggest that blood flow directly acts on endothelial cells during blood vessel pruning, since pruning rapidly resumed after reestablishing the heartbeat. We note that in embryos with a global block in blood flow, CrDI pruning still occurred in about 40% of analyzed embryos, suggesting the existence of flow independent mechanisms controlling blood vessel pruning. The identification of these mechanisms and their potential integration with flow-based processes will greatly advance our understanding of blood vessel pruning and vascular remodeling.Previous studies showed that pruning of early forming hindbrain vessels in zebrafish embryos occurs normally in the absence blood flow s843Tg(kdrl:EGFP)s916Tg(kdrl:Hsa.HRAS-mCherry)y7Tg(fli1a:nEGFP)sd2Tg(gata1a:DsRed)Tg(UAS:RFP) (kindly provided by Kawakami lab), zdf13Tg(HSP:Cre)ubs12Tg(UAS:VE-cadherin-deltaC-EGFP)mu126Tg(kdrl:cytobow1.0) and mu122Tg(kdrl:H2B-EGFP) (this study).Zebrafish were maintained as described previously Danio rerio Histone2B (H2B) fused in frame to EGFP was used to PCR amplify and subclone H2B-EGFP into pBluescript. Thereafter, H2B-EGFP was cloned via EcoRV and EcoRI into pKdrl:MCS-Tol2 tol2 transposase mu122Tg(kdrl:H2B-EGFP).pCS2-H2B-EGFP mu126Tg(kdrl:cytobow1.0) fish, we used NheI and DraIII to release the cassette containing the fluorescent proteins from CMV Brainbow 1.0 \u201cL\u201d (Addgene 18721) mu126Tg(kdrl:cytobow1.0). Cytobow 1.0L constructs contain two floxed fluorescent proteins (tdTtomato and Cerulean) and Yellow Fluorescent Protein. Cre-mediated recombination between individual Lox-P sites can lead to the expression of different combinations of fluorescent proteins, as previously shown mu125Tg(kdrl:cytobow1.0) fish to zdf13Tg(HSP:Cre) fish. This resulted in 25% double transgenic offspring. Embryos were subsequently heat shocked at 90% epiboly and those with differently labeled endothelial cells in anterior blood vessels were chosen for time-lapse analysis.In order to generate s843Tg(kdrl:EGFP) embryos to analyze the regression of the dorsal CrDI. Regression was analyzed at 48 hpf under a stereomicroscope or via confocal microscopy. Only embryos with normal blood flow patterns at 36 hpf were included in the analysis. We defined regression as complete loss of endothelial connection between CrDI and PMBC. Each experiment was performed in triplicates with 20 eyes being analyzed per experiment. Quantification charts display the average values of three experiments. Data were evaluated with two-tailed unpaired Student\u2019s t test and significance was calculated for regression.We used ubs12Tg(UAS:VE-cadherin-deltaC-EGFP) line was performed as previously described in vivo imaging, live embryos were mounted essentially as described previously mu122Tg(kdrl:H2B-EGFP) disappeared in the ablated vessel. After NCA ablation, we monitored the embryos for PMBC integrity and normal blood flow.Live imaging of endothelial cell junctions using the 2O. Embryos were injected at one cell stage with 4 ng MO tnnt2a (silent heart 5\u2032 CATGTTTGCTCTGATCTGACACGCA 3\u2032) pu.1 (5\u2032-GATATACTGATACTCCATTGGTGGT-3\u2032) 5\u2032-CCTCTTACCTCAGTTACAATTTATA-3\u2032). Danio rerio bcl2 mRNA was transcribed from pCS2-Bcl2 (kindly provided by J. Hillmer) using Message Machine kit (Ambion) and injected into one cell stage with a final amount of 100 pg.MO were obtained from Gene-Tools and dissolved in dHFor AO staining, embryos were incubated for 30 min in 10 \u00b5g/ml AO in E3 at 28.5\u00b0C, followed by washing for 30 min. Drug treatments were carried out on dechorionated embryos in E3 medium. To completely stop the heart at various time points, we either treated zebrafish embryos with 4X Tricaine or with 2X Tricaine in addition to 2.5 \u00b5M Nifedipine File S1Supporting tables.(DOC)Click here for additional data file.Movie S1(Related to): Time lapse imaging of early eye blood vessel development. Time-lapse imaging of s916Tg(kdrl:Hsa.HRAS-mCherry); y7Tg(fli1a:nEGFP) embryos. Images were taken from 32\u201348 hpf at 15 minute time intervals. Endothelial cell membranes are marked by red fluorescence, while endothelial cell nuclei are marked in white. Arrows indicate the forming Nasal Ciliary Artery (NCA). After connection of the NCA (indicated by arrows) to the Cranial Division of the internal carotid artery (CrDI), the dorsal CrDI prunes (arrowheads).(MOV)Click here for additional data file.Movie S2Cellular rearrangements during CrDI pruning based on labeling of individual endothelial cells (Related to). Time-lapse imaging of mu126Tg(kdrl:cytobow 1.0); zdf13Tg(HSP:Cr) embryos. Images were taken from 34.5\u201340 hpf at 15 minute time intervals. Distinct endothelial cells were false colour labeled in green and magenta, respectively. Note absence of magenta cell in dorsal CrDI during unicellular configuration prior to CrDI pruning.(MOV)Click here for additional data file.Movie S3Cellular rearrangements during CrDI pruning based on labeling of endothelial cell junctions (Related to). Time-lapse imaging with ubs4Tg(UAS:RFP); Tg; ubs12Tg(UAS:VE-cadherin-deltaC-EGFP) in a frontal-lateral orientation between 36 and 45 hpf. In the overlay channel, endothelial cells are labeled in red and endothelial junctions are shown in green. Middle panels show junctional labeling alone with individual cells and junctions colour coded to illustrate junctional dynamics during CrDI pruning. Right panel shows RFP channel alone, marking endothelial cell bodies.(MOV)Click here for additional data file.Movie S4(MOV)Click here for additional data file.Movie S5(MOV)Click here for additional data file.Movie S6Laser ablation of the NCA and influence on CrDI pruning (Related to). Laser ablation of the NCA was carried out at 32 hpf and time-lapse imaging was performed from 34\u201348 hpf in mu122Tg(kdrl:H2B-EGFP), s916Tg(kdrl:Hsa.HRAS-mCherry) double transgenic zebrafish, labeling all endothelial nuclei in green and endothelial membranes in red. In the absence of a functional NCA, the CrDI fails to prune.(MOV)Click here for additional data file."} +{"text": "Endothelial progenitor cells (EPCs) are thought to exert beneficial effects on atherosclerosis, angiogenesis, and vascular repair. We performed a randomized pilot \"proof of concept\" study of traditional acupuncture (TA) and circulating EPCs in patients with coronary heart disease (CHD).Thirteen subjects were randomized to TA, sham acupuncture (SA) or waiting control (WC) for 12 weeks. TA received treatments at CHD specific acupuncture points while SA received tube pressure with no needle insertion proximate to the TA site but not considered active. WC received no intervention. EPCs at study entry/exit were quantified by flow cytometry using CD34, CD133 and VEGFR2 cell surface markers.Eight men and five women (mean age 59\u00b111 yrs) were included. Compared to entry, TA had a significant increase in percentage change from baseline in level of cells expressing CD34+/VEGFR2+ compared to SA and WC (p=0.03). No group differences were evident in immature EPCs expressing CD34+/CD133+, although these were lowest in the TA group; numbers did not correlate with elevation in the CD34+/VEGFR2+group.Results suggest that TA may increase mobilization of specific EPC sub-populations (VEGF R2) while simultaneously decreasing levels of a more immature cell type (CD133) and may beneficially alter levels of EPCs. This pilot study provides feasibility and outcome variability data for clinical trial planning purposes. Further studies are warranted to evaluate whether traditional acupuncture can beneficially impact CHD via augmentation of EPC regenerative pathways."} +{"text": "The association between viral infections such as hepatitis C virus (HCV) and autoimmunity has been proposed. Antibodies against HCV have been demonstrated in sera from up to 13% adult patients with SLE, however data regarding juvenile onset SLE (jSLE) is lacking despite potential increased risk factors associated.To determine the prevalence and possible association of HCV infection and jSLE.Methods: We evaluated 40 jSLE patients (according to ACR criteria); mean age = 19 \u00b1 4.4 years and mean disease duration = 6 \u00b1 3.2 years; 34 females, 6 males. Twenty healthy children and 20 rheumatic fever patients matched for age and sex were included as controls. All subjects were interviewed in order to search for risk factors for HCV infection including use of endovenous drugs, blood products transfusions, promiscuous sexual activity, previous hospitalizations and/or invasive diagnostic or therapeutic procedures. Serum samples were tested for ANAs by standard techniques and for anti-HCV antibodies using a high sensitive third generation microparticle enzyme immunoassay .Thirty-six (90%) jSLE patients were under immunossupressive therapy; 47 hospitalizations and 19 invasive procedures were required by jSLE subjects contrasting to only 5 hospitalizations and 5 invasive procedures by the control group. ANA titers were elevated in all 40 jSLE sera and negative in all controls. Remarkably only one (2.5%) jSLE sera was anti-HCV positive compared to all uniformly negative control sera.Contrary to adult onset SLE, the low prevalence of anti-HCV antibodies in sera from jSLE patients suggests lack of association of HCV infection and jSLE."} +{"text": "In humans, gender difference has been suggested by previous studies. Sex difference in vitamin C requirements have also been reported from animal models. Mouse spontaneous fracture model (sfx) lacks L-gulonolactone oxidase (LGO), an essential gene for the synthesis of ascorbic acid (Vitamin C). We have been using sfx model for the past eight years and we are the first group to identify the LGO mutation in this mouse model.In order to identify the genes that regulate skeletal development through Vitamin C (VC) pathway, we analyzed the gene expression profile in mouse femur. For microarray analysis, femur from three age-matched, wild-type +/+ Balb/By, inbred strain mice (WT) and 3 female and 3 male homozygous sfx/sfx mice were used.Myogenic factor 6 (Myf6) gene is differentially regulated by VC in male versus female femurs while Myf5 and Pax6 are almost undetectable in the mouse femur. Expression of Myf6 in female mice measured by DNA microarray experiment Myf6 is much down regulated. Myf6 is up regulated in femurs of male sfx mice. Myf6 is specifically expressed in skeletal system (femur) while in liver and kidney Myf6 is not expressed. There are sex differences in expression levels of collagen synthesis genes. There are sex differences between expression levels of hormone relevant genes. The differential expression of Myf6 was confirmed by real time PCR.Vitamin C may differentially regulate myf6 and collagen expression in the skeletal development through sex hormones and relevant factors."} +{"text": "Vacuum-assisted biopsy (VAB) systems have been shown to outperform 14G core needle biopsy (CNB) reducing the need for diagnostic or multiple surgeries. We introduced VAB in 2011 with the aim of reducing our B3 rate and increasing rate of preoperative diagnosis of invasive cancer.Mammographic abnormalities requiring stereotactic biopsy in a defined period which had either 14G CNB or VAB were included. Data collated included mammographic findings, biopsy and surgical outcome.Ninety 14G-CNBs (November 2010 to May 2011) were compared with 78 VABS (July 2012 to January 2013). There was no reduction in our B3 rate (12 and 13 cases respectively) although VAB had higher sensitivity with better correlation between biopsy diagnosis and surgical histology (77% compared with 42%). Four of 12 (25%) B3 lesions diagnosed following 14G CNB were upgraded to DCIS following surgical excision compared with no cases following VAB. Preoperative invasive malignancy diagnostic rates were unchanged, 15% of cases were upgraded in both groups.VAB has not reduced our B3 rate but has increased the accuracy of our preoperative diagnosis of non-invasive cancer supporting current theories that patients who have undergone vacuum-assisted biopsy may not require surgical diagnostic excision. However, we recognise the number of cases is low and further cases will be analysed and presented."} +{"text": "Adequate anticoagulation is essential to achieve efficient and cost-effective renal and liver replacement therapy. However, critically ill patients with advanced liver cirrhosis are associated with low antithrombin III (ATIII) serum levels and increased tendency to both coagulation and bleeding disorder. Thus, we hypothesized that single-dose application of antithrombin III prolongs filter lifetime during renal and liver replacement therapy in critically ill patients with advanced liver cirrhosis without causing additional bleeding problems.In this retrospective study, data of 33 extracorporeal therapies in nine critically ill patients with advanced liver cirrhosis admitted to a medical ICU in 2007 and 2008 were analyzed. Included patients underwent either continuous renal replacement therapy (CRRT), intermittent hemodialysis (IHD) or liver replacement using the molecular adsorbents recirculation system (MARS) with single doses of ATIII as sole anticoagulant. Bleeding complications and filter lifetimes were used as outcome parameters.Data were available for 13 CRRT, 14 IHD, and six MARS filters with total filter lifetimes of 661 (CRRT), 66 (IHD), and 42 hours (MARS), respectively. Mean filter lifetimes were 44.0 \u00b1 27.9 (CRRT), 4.7 \u00b1 1.6 (IHD), or 4.6 \u00b1 12.6 hours (MARS). Fifteen percent (two out of 13) of CRRT filters, 7% (one out of 14) of IHD filters and 0% (zero out of six) of MARS filters were lost due to clotting of the dialysis circuit. New onset of bleeding was not observed during IHD, MARS and CRRT.Our data suggest that single-dose application of ATIII is effective and safe as alternative anticoagulation in critically ill patients with advanced liver cirrhosis. However, prospective controlled trials are necessary to confirm our findings."} +{"text": "Members of the mammalian sirtuin family, Sirt1 \u2013 Sirt7, are known to regulate metabolic processes especially carbohydrate and fat metabolism . Sirt1 aTo discover new regulators of Sirt1 activity we performed an unbiased screen for molecules that might interact with Sirt1 using a label free quantitative mass spectrometry based co-immunoprecipitation strategy. We identified Sirt7 as a novel Sirt1 binding protein. The interaction between Sirt1 and Sirt7 was confirmed by immunoprecipitation of endogenous proteins and GST pull-down assays. Sirt1 protein expression and enzymatic activity was increased in WAT of Sirt7 knockout mice leading to age-dependent lipodystrophic phenotype. Increased Sirt1 activity might account for resistance of Sirt7 knockout mice fed high fat diet against liver steatosis. In vitro experiments demonstrated a diminished ability of Sirt7 deficient MEFs and primary preadipocytes to undergo adipogenesis. These defects were rescued by knock-down of Sirt1 or in cells deficient for one Sirt1 allele .Our results highlight the importance of cross-regulatory circuits among individual members of the sirtuin family in organismal homeostasis. Lack of Sirt7 leads to a sustained activation of Sirt1. Apparently, such un-physiologically exaggerated, persistent Sirt1 activation results in metabolic dysfunction and nullifies its principally beneficial effects such as fat mobilization and inhibition of adipogenesis."} +{"text": "The mitochondrial NAD fasting and repr fasting . We sougOur approach was two-pronged: using the DNA sequence analysis program PhylCRM , we analWe have identified candidate transcription factors that may affect Sirt3 expression levels, including the zinc finger transcription factor MZF1. We have also analyzed the effect of several drugs on Sirt3 expression, notably observing a decrease in Sirt3 expression with resveratrol treatment.We have identified transcription factors and calorie restriction mimetic drugs which may control Sirt3 expression and are currently conducting follow-up studies."} +{"text": "We defined a case-patient as a patient with culture of V. vulnificus from any clinical specimen. A record search of clinical case notes was performed through a computerized clinical management system maintained by the Hospital Authority, which manages all public hospitals in Hong Kong. For each case-patient identified, we reviewed demographic data , clinical and laboratory data , and potential risk factors associated with the case. We compared previously healthy patients with patients who had predisposing medical conditions in terms of demographic profile, clinical signs and symptoms and outcome, and known exposure factors. Mann-Whitney U tests, \u03c7We identified 29 cases over the 32-month study period. Twenty-two (76%) patients had disease onset from May through August, the summer season in Hong Kong. Fifteen (52%) cases were in men, and the median age was 70 years (range 24\u201382 years). Fifteen (52%) patients had underlying illnesses that were known to predispose them to V. vulnificus infection, including chronic liver disease (30%), chronic renal failure (15%), diabetes mellitus (7%), and thalassemia major (3%). Fourteen (48%) patients were previously healthy. No significant differences in age and sex were found.Among the 14 previously healthy patients, the consequences of V. vulnificus infection included necrotizing fasciitis (70%), severe cellulitis (7%), primary septicemia (14%), and gastroenteritis (7%). Two patients who had necrotizing fasciitis and 1 patient with primary septicemia died. Compared with patients with predisposing medical conditions, patients with a history of good health had a higher (but not significant) proportion of necrotizing fasciitis , a lower proportion of septicemia , and an equal number of severe cases of cellulitis (7% vs. 7%). Furthermore, fewer patients with a history of good health died than did patients with predisposing illnesses . The median duration between symptom onset and admission for all patients was 1 day (range 0\u20133 days), with no significant difference between the 2 groups.A history of cutaneous injury or a skin prick from a seafood part was significantly more common among previously healthy patients than among patients with predisposing illnesses . Ten (83%) of the 12 previously healthy patients with necrotizing fasciitis and septicemia reported a history of cutaneous injury. The corresponding proportion was significantly lower (31%) among patients with predisposing medical conditions (p = 0.01). Among all 29 patients, a history of eating raw oysters or other raw or undercooked seafood before illness onset was uncommon and was only reported by 1 patient. Although V. vulnificus has not been proven as the cause of gastroenteritis, Hseuh et al. have suggested that such results might have occurred because patients with diarrhea seldom sought care from a large teaching hospital or saved stool samples for investigation (\u2013,\u2013V. vulnificus infection was first reported in humans in 1979 ("} +{"text": "RAD51D have been associated with an increased risk of hereditary ovarian cancer and although they have been observed in the context of breast and ovarian cancer families, the association with breast cancer is unclear. The aim of this current study was to validate the reported association of RAD51D with ovarian cancer and assess for an association with breast cancer. We screened for RAD51D mutations in BRCA1/2 mutation-negative index cases from 1,060 familial breast and/or ovarian cancer families (including 741 affected by breast cancer only) and in 245 unselected ovarian cancer cases. Exons containing novel non-synonymous variants were screened in 466 controls. Two overtly deleterious RAD51D mutations were identified among the unselected ovarian cancers cases (0.82%) but none were detected among the 1,060 families. Our data provide additional evidence that RAD51D mutations are enriched among ovarian cancer patients, but are extremely rare among familial breast cancer patients.Mutations in RAD51D/RAD51L3; MIM#602954) is a component of the homologous recombination DNA repair pathway. The RAD51D protein forms a protein complex with RAD51B, RAD51C and XRCC2 that binds to single stranded DNA (including single stranded gaps in double stranded DNA) and is required for the formation of RAD51 foci in response to DNA damage RAD51D among 911 families with histories of breast and ovarian cancer, compared to one mutation among 1,060 population controls. They reported a significantly elevated risk of ovarian cancer but did not detect a significantly elevated risk of breast cancer . They also reported that mutations are more prevalent in multiple case ovarian cancer families. RAD51D has subsequently been investigated in an additional series of 175 breast and ovarian cancer families, with an additional mutation being identified among the 51 families with at least two ovarian cancers (and among the 75 probands affected by ovarian cancer) RAD51 homolog D (S. cerevisiae) (BRCA1 and BRCA2 were the only genes known to confer a considerable risk of ovarian cancer (in conjunction with breast cancer) with two recent studies reporting that 13.3\u201314.1% of unselected high grade ovarian cancers are accounted for by mutations in one of these two genes RAD51CRAD51D mutations. To validate the association of RAD51D mutations to ovarian cancer and assess if there is any risk for breast cancer risk, we screened all coding exons in germline DNA from an unselected cohort of 245 unselected ovarian cancer patients and BRCA1/2-unrelated index cases from 1,060 breast and/or ovarian cancer families. Exons containing novel, non-synonymous variants among these cases were screened in a panel of 466 cancer-naive control samples.Until recently, The unselected ovarian cancer cohort included 245 individuals with various histological subtypes of ovarian cancer . These samples were obtained from patients presenting to hospitals in the south of England, UK BRCA1 or BRCA2 mutation having been identified. The ethnicity of the index cases was self-reported as Caucasian in the vast majority of cases. All individuals provided written, informed consent for genetic testing of the genetic causes of hereditary breast and ovarian cancer and subsequently tested negative for mutations in BRCA1 and BRCA2. This study was approved by the Peter MacCallum Cancer Centre Human Research Ethics Committee. In total, index cases from 1,060 families were examined in this study, including 16 with a family history of ovarian cancer only, and 303 with a family history of both breast and ovarian cancer. Of these index cases, 98 had a personal history of ovarian cancer. The remaining 741 families had a personal and family history of breast cancer only.The familial cohort included 540 individuals with verified personal and family histories of breast and/or ovarian cancer who were previously assessed at the Peter MacCallum Cancer Centre Familial Cancer Centre , as well as index cases from 520 multiple case breast cancer families (with or without ovarian cancer) obtained from the Kathleen Cunningham Foundation Consortium for Research into Familial Breast Cancer (kConFab) Cancer-naive control DNA samples were obtained from kConFab (231 age- and ethnicity-matched best friend controls) and from the Princess Anne Hospital, UK RAD51D with amplicons ranging in size from 215\u2013277 bp for high resolution melt (HRM) analysis (www.hgvs.org/mutnomen). All novel variants were verified by Sanger resequencing of non-WGA DNA. Tumour cells were needle dissected from 10 \u00b5m sections to obtain tumour DNA, which was subsequently whole genome amplified.DNA for mutation screening underwent whole genome amplification (WGA) using the Repli-G amplification system (Qiagen). Ten primer pairs were designed to amplify the ten coding exons of analysis . HRM anain silico prediction tools were used to assess the likely functional effect of the missense variants identified in this study: PolyPhen-2 The following RAD51D mutations (i.e. high grade serous and endometrioid) is consistent with the majority of mutations reported in other RAD51D studies e.g. BRCA1 and BRCA2), but the number of mutations in RAD51D identified to date is too few to determine the significance of this observation. A third truncating mutation, p.(Lys91Ilefs*13), was identified in one of 466 control samples (0.21%). All three of these mutations have previously been reported Two previously reported truncating mutations, p.(Arg186*) and p.(Trp268*) were identified among a series of 245 unselected ovarian cancer patients (0.82%). The p.(Arg186*) variant was detected in a patient diagnosed with a grade 2 papillary serous cystadenocarcinoma at 66 years of age. DNA sequence analysis of tumour tissue obtained from this tumour showed reduction of the wildtype allele consistent with loss of heterozygosity (LOH) . The p. nonsynonymous variants were detected, once each among 741 breast only cancer families. Three of these variants were novel: p.(Met16Thr), p.(Gly96Cys) and p.(Arg266Cys); the remaining two variants, rs150498754 and rs140285068, are reported in the Exome Variant Server (EVS) database at frequencies of <0.02%. In silico analysis tools predicted that variants p.(Gly96Cys) and p.(Arg266Cys) would likely affect protein function is in keeping with that (0.6%) estimated by Loveday et al., and higher than observed in controls (0.21%). However, it is possible that the variant frequency reported here could be an underestimate due to the reduced sensitivity of HRM analysis compared to direct resequencing . Five of the variants detected in this study have previously been reported in RAD51D mutation studies by Loveday et al. or Osher et al. http://evs.gs.washington.edu/EVS/) [June 2012]).The frequency of truncating germline RAD51D mutations do not contribute significantly to breast cancer risk.The absence of truncating mutations in 741 breast cancer only families (or 962 breast cancer-affected probands) provides further evidence that"} +{"text": "Introduction. To evaluate whether secondary resection of lymph node (LN) metastases (LNMs) can result in PSA remission, we analysed the PSA outcome after resection of LNM detected on PET/CT in patients with biochemical failure. Materials and Methods. 11 patients with PSA relapse after radical prostatectomy without adjuvant therapy were included. Suspicious LN (1\u20133) detected on choline PET/CT and nearby LN were openly dissected (09/04\u201302/11). The PSA development was examined. Histological and PET/CT findings were compared. Results. 9 of 10 patients with histologically confirmed LNM showed a PSA response. 4 of 9 patients with single LNM had a complete permanent PSA remission . Of metastasis-suspicious LNs (14) 12 could be histologically confirmed. The additionally removed 25 LNs were all correctly negative. Conclusions. The complete PSA remissions after secondary resection of single LNM argue for a feasible therapeutic benefit without adjuvant therapy. For this purpose the choline PET/CT is in spite of its limitations currently the most reliable routinely available diagnostic tool. In prostate cancer the long relapse-free survival of patients with 1-2 LNM even without adjuvant therapy in the primary situation [11C]choline PET/CT without adjuvant therapy [18F]fluoroethylcholine were included too.In our first studies of PET/CT-guided secondary LN surgery, we reported on the outcome of all in all 8 patients with LNM detected by using choline and [18F]fluoroethylcholine PET/CT provides the opportunity to detect small LNM (>5\u2009mm) in prostate cancer with exact topographic allocation and so the targeted resection of LNM. In contrast, the computed tomography (CT) and the conventional magnetic resonance imaging (MRI) are not applicable for early detection of LN recurrence. The lymphotropic nanoparticle-enhanced MRI can detect smaller LNM (>2\u2009mm) [The integrated [ine and 1Ffluoroet11C]choline PET/CT (n = 9) or [18F]fluoroethylcholine PET/CT (n = 2) in case of PSA failure were included. All had a PSA increase or persistence after operative therapy which was performed between 3 months and 9 years ago. In 10 patients a radical retropubic prostatectomy with pelvic LN dissection (PLND) and in one patient only a radical retropubic prostatectomy were carried out. One patient had received a sentinel guided PLND (sPLND) on both sides of the pelvic and also an extended PLND (ePLND) on the right side because of an advanced tumor and another one only sPLND in our clinic. The remaining patients had received conventional PLND, carried out by other institutions. There had to be negative margins and no clue for a local relapse or distant metastasis. The patients were informed that there is no conclusive data concerning survival benefit after secondary LN surgery in written and oral form, and signed an informed consent. 11 consecutive patients with 1\u20133 LNM detected by using [11C]choline or [18F]fluoroethylcholine PET/CT studies were performed with integrated PET/CT systems externally in four centres with a high level of expertise. Experienced radiologists and nuclear medicine specialists evaluated the images to anatomically localize the sites of pathologic choline uptake. The diagnosis of tumor positive LN on PET/CT images was based on the visual evidence of the presence of focal increased choline uptake on PET images, whose location corresponded to LN on CT images (All choline or [18F]fluoroethylcholine PET/CT is, despite its limitations, currently the most reliable routinely available diagnostic tool. Whether the secondary resection of LNM has an influence on the course of disease or could even be curative must be demonstrated in further studies in consideration of more patients and a long-term followup. Especially patients with minimal LNM seem to benefit from the secondary removal of LNM in prostate cancer. The here observed lasting, complete PSA remissions after secondary resection of single LNM appear to have a feasible therapeutic benefit without adjuvant therapy. For this purpose, the ["} +{"text": "Cardiopulmonary bypass (CPB) is regarded as an important contributor to acute kidney injury and use of renal-dose dopamine to protect the kidneys against hypoperfusion injury following cardiac surgery remains controversial. Cystatin C has been described as a sensitive biomarker of early renal tubular injury. We aimed to evaluate the effect of renal-dose dopamine on renal tubular functions in patients undergoing coronary artery bypass grafting (CABG) surgery.n = 19) or saline as placebo starting from induction of anesthesia for 48 hours. Serial blood and urine samples after induction of anesthesia and 2, 12, 24, 48 hours post CPB were collected to measure serum cystatin C, creatinine levels and urinary \u03b22-microglobulin. Intraoperative and daily measurements of hemodynamic parameters and urine output were recorded.Thirty-six patients undergoing CABG surgery were prospectively randomized to receive either 2 \u03bcg/kg/minute dopamine infusion . See Table 2-microglobulin levels between the groups (P > 0.05 for both). GFR was preserved equally in both groups on postoperative day 2 .The groups were similar in terms of physical characteristics, perioperative hemodynamic measurements, urine outputs and surgical times. Serum cystatin C levels demonstrated similar increases during 12, 24 and 48 hours post CPB in the dopamine and placebo groups (The results suggest that renal-dose dopamine does not exacerbate the severity of renal tubular injury when compared with the untreated controls during the early postoperative period of patients undergoing CABG surgery."} +{"text": "Germline development is characterized by genome-wide reprogramming of DNA methylation. Recent work has enlightened the dynamics of DNA methylation in primordial germ cells (PGCs), but knowledge of histone modification dynamics at these developmental stages remains limited, mostly due to the difficulty in obtaining enough high quality chromatin immunoprecipitation (ChIP) material for sequencing. Previous work in our laboratory has demonstrated the importance of histone methyltransferases in silencing retroelements and a suWe have developed a scaled down native ChIP and sequencing library construction protocol that can be performed on small cell numbers. We optimized sample fragmentation, antibody concentration, ChIP conditions, library construction and amplification to generate high quality, high resolution H3K9me3 sequencing libraries from as little as 1,000 embryonic stem cells. Paired-end sequencing (Illumina HiSeq) of these pooled and indexed libraries generated an average of 28 million aligned read pairs (with 6 libraries per sequencing lane), with under 20% duplicate reads, for an average of 23 million unique read pairs. Under optimized conditions, we found that over 85% of the peaks identified using standard native ChIP-sequencing were also detected by our small cell number native ChIP-seq protocol. We also found excellent reproducibility between independent ChIP experiments. Using this optimized small cell number native ChIP-seq protocol, we generated genome-wide H3 and H3K9me3 profiles from 1,000 E13.5 PGCs and identified a unique cohort of genes and retroelements enriched for this repressive mark. Integration of this chip-seq data with DNA methylation/WGBS and transcriptome data generated at the same developmental stage will be presented."} +{"text": "Alpinia calcarata conducted in this study identified the presence of an unusual intron at the region forming the second exon of typical PKSs, forming a gateway information of distribution of novel PKSs in Zingiberaceae.Plant phenolic compounds form a valuable resource of secondary metabolites having a broad spectrum of biological activities. Type III polyketide synthases play a key role in the formation of basic structural skeleton of the phenolic compounds. As a group of medicinal plants, PKSs with novel features are expected in the genome of Zingiberaceae. The genomic exploration of PKS in The Type III polyketide synthase (PKS) superfamily of enzymes play an important role in the biosynthesis of phenolic compounds of diverse structure and function in plants . ChalconAlpinia calcarata, to unveil its genomic features.Polyketide synthases have highly conserved stretches of amino acid residues forming the catalytic pockets, and the residues forming these pockets are located mainly within the second exon , 1.5 mM MgCl2 and 200 \u03bcM of each dNTPs. Amplifications were carried out in a BioRad iCycler using the following conditions: initial denaturation for 5 min at 94 \u00b0C, 35 cycles of 94 \u00b0C for 30 s, 65 \u00b0C for 1 min and 72 \u00b0C for 1 min, followed by a final extension at 72 \u00b0C for 7 min. The PCR products were analyzed on 2% agarose gel and purified using the GFX gel band purification kit (Amersham). The purified PCR product was ligated into pGEM-T Easy plasmid vector (Promega). The ligation reaction mix (10 \u03bcL) contained 1X ligation buffer, 50 ng of PCR product, 50 ng of pGEM-T Easy vector DNA and 3 units of T4 DNA ligase. The ligation was carried out overnight at 4 \u00b0C and the product was used for transformation. Positive transformants were selected by blue white screening and colony PCR. Plasmid isolation was carried out from positive colonies by alkaline lysis , multiple sequence alignments with selected PKSs were run using the Clustal W program. For this the sequences were first aligned at the amino acid level and subsequently, the nucleic acid sequences were aligned according to the amino acid sequences. Intron prediction was done by using the intron prediction program at NetPlantGene Server.Aloe arborescens PCS (AY823626), Aloe arborescens OKS (AY567707), Arabis alpina CHS (AF112084), Arachis hypogaea RES (DQ124938), Arabidopsis thaliana CHS , Callistephus chinensis CHS (Z67988), Camellia sinensis CHS (D26593), Cardamine amara CHS (AF112085), Daucus carota CHS (AJ006780), Dictamnus albus CHS (AJ850132), Escherichia coli fabH (M96793), Gerbera hybrida 2-PS (Z38097), Humulus lupulus VPS (AB047593), Hordeum vulgare CHS (X58339), Hydrangea macrophylla CTAS (AB011468), Hypericum androsaemum BPS (AF352395), Ipomoea purpurea CHS (AB001826), Ipomoea batatas CHS (AB037389), Lilium hybrid CHS (AF169798), Medicago sativa CHS (L02901), Oryza sativa CHS (X89859), Petunia hybrida CHS (X14593), Phalaenopsis sp. \u2018pSPORT1' BBS (X79903), Phaseolus vulgaris CHS (X06411), Pinus strobus CHS (AJ004800), Pisum sativum CHS (X63335), Psilotum nudum STS (AB022685), Rheum palmatum BAS (AF326911), Rheum palmatum ALS (AY517486), Rorippa amphibia CHS (AF144530), Ruta graveolens ACS (AJ297786), Sorghum bicolor CHS (AY069951), Triticum aestivum CHS (AY286098), Vitis vinifera STS (EF192465), Wachendorfia thyrsiflora PKS (AY727928), Zingiber officinale PKS (DQ486012), Brassica napus CHS (AF076333), Dendrobium nobile CHS (ABE77392), Iris germanica CHS (BAE53636), Curcuma longa CURS3 (AB506763), Curcuma longa CURS2 (AB506762), Curcuma longa CURS1 (AB495007), Curcuma longa DCS(AB495006), Oryza sativa CUS (AK109558) and Streptomyces griseus RppA (AB018074).Phylogenetic analysis was carried out using the Neighbour Joining (NJ) and Maximum Parsimony (MP) methods implemented in MEGA3 showed a highest identity of 87% to the PKS/CHS of Zingiber officinale, with the core fragment coinciding with the region between 546-1101 bp of typical PKS/CHS. Type III PKSs in plants generally have a size of 1170 bp, coding for 390 amino acids. The most characteristic feature of the core region is its location in the second exon, which contains most of the residues of the catalytic region. Comparative analysis of PKS/CHS sequences from all the plants revealed that all of them have only one intron, except for Antirrhinum majus to Ser, Ser (338) to Gln, Ileu (254) to Val, when compared to typical CHSs. The numbering and type of amino acid residues is based on the amino acid sequence of M. sativa CHS. These changes observed on A. calcarata PKS can be taken as an indication of function(s) other than typical chalcone formation. The presence of the unusual 93 bp intron in A. calcarata may be of significance to the evolution of the PKS gene family in Zingiberaceae. All CHS genes studied so far contained an intron at a conserved position, but in Antirrhinum majus a second intron was found to be present in the second exon (Physcomitrella patens (Polygonum cuspidatum (Use of PKS specific primers for the PCR resulted in the formation of a 703 bp product from genomic DNA of alcarata . This amum majus . The firum majus . The intond exon . Very reRheum palmatum, a phylogenetic analysis including four members of the PKS superfamily generated separate clusters for CHSs and non CHSs. Such separate clustering was also reflected in the biochemical activities (The phylogenetic analysis of AcPKS with other PKS sequences showed separate clustering, distinct from typical CHS/PKSs and along with recently identified plant-specific nonchalcone forming PKSs . The sepA. calcarata PKS identified in this study shows the unusual occurrence of an intron that can be considered as a novelty. The sequence identified in this study opens a perspective for further molecular exploration of the PKS family in A. calcarata.Most of the genes coding for the biosynthetic machinery of plant secondary metabolism are encoded by small families of genes originated through duplication over evolutionary time . In legu"} +{"text": "Escherichia coli are a serious problem for the dairy industry. Reduction of incidence and severity of mastitis is desirable and mild priming of the immune system either through vaccination or with low doses of immune stimulants such as lipopolysaccharide LPS was previously found to dampen detrimental effects of a subsequent infection. Monocytes/macrophages are known to develop tolerance towards the endotoxin LPS as adaptation strategy to prevent exuberant inflammation.Udder infections with environmental pathogens like We have recently observed that infusion of 1 \u03bcg of LPS into the quarter of an udder effectively protected for several days against an experimentally elicited mastitis. We have modelled this process in primary cultures of mammary epithelial cells (MEC) from the cow. MEC are by far the most abundant cells in the healthy udder coming into contact with invading pathogens and little is known about their role in establishing ET.7/ml particles of heat inactivated E. coli bacteria for six hours. Priming-related alterations in the global transcriptome of those cells were quantified with Affymetrix microarrays. LPS priming alone caused differential expression of 40 genes and mediated significantly different response to a subsequent E. coli challenge of 226 genes. Expression of 38 genes was enhanced while that of 188 was decreased. Higher expressed were anti-microbial factors , cell and tissue protecting factors as well as mediators of the sentinel function of MEC . Dampened was the expression of potentially harmful pro-inflammatory master cytokines and immune effectors . Functional network analysis highlighted the reduced expression of IL1B and of IRF7 as key to this modulation.We primed primary MEC cultures for 12 h with LPS (100 ng/ml) and stimulated three cultures either 12 h or 42 h later with 10LPS-primed MEC are fitter to repel pathogens and better protected against misguided attacks of the immune response. Attenuated is the exuberant expression of factors potentially promoting immunopathological processes. MEC therefore recapitulate many aspects of ET known so far from professional immune cells. Escherichia coli (E. coli) often provokes severe inflammation, greatly reduces milk yield and may eventually cause heavy tissue damage in the mammary gland , receptor transporter protein 4 [RTP4], 2'-5'-oligoadenylate synthetase 1 [OAS1], interferon stimulated exonuclease gene 20 kDa [ISG20]). The factor SLPI has not only a broad spectrum of antimicrobial activities but also anti-inflammatory/immunomodulatory functions . Background correction and normalization of the expression values was performed using the GC Robust Multi-array Average (GCRMA) algorithm [vs. control cultures. Human orthologs of differentially expressed genes (DEG) were determined as described previously [The microarray data were analysed using the Biometric Research Branch (BRB) array tools version 4.1 [lgorithm . Transcreviously ,52. Geneeviously .6 to 10 copies) of those sequenced plasmids. Sequences of the oligonucleotide primers used are given in Additional file Total RNA extraction with Trizol , cDNA preparation with Superscript II (Invitrogen) and quantification of mRNA copy numbers with the LightCycler instrument and the SYBR Green plus reagent kit was as described . QualityJG contributed to designing the experiments, analyzed and interpreted the data, and drafted the manuscript. WP contributed to the experimental design, data analysis, and drafting the manuscript. HZ, HJS and LG contributed to conception of the study and critical data interpretation. DK prepared the microarray hybridizations. HMS designed and supervised the study and helped drafting the manuscript. All authors read and approved the final manuscript.versus Control (C.)Table S1: All DEG from comparison Priming (P.) . A) Short time waiting experiment (40 IPA mapped DEG). B) Long time waiting experiment (13 IPA mapped DEG)Click here for fileversus Induction (I.)Table S2: All DEG from comparison Induction post Priming (I.p.P.) . A) Short time waiting experiment (226 IPA mapped DEG). B) Long time waiting experiment (6 IPA mapped DEG)Click here for fileTable S3: Comparison of RT-qPCR and microarray measurements of selected candidate genesClick here for fileTable S4: All RT-qPCR values (relative mRNA copy numbers) contributing to Figure 4.Click here for fileTable S5: Correlation of relative mRNA concentrations determined in microarray hybridizations or RT-qPCR from three biological pbMEC replica of the four challenge groups of the short and long waiting experimentsClick here for fileTable S6: Sequences of oligonucleotide primers used for real-time PCR quantificationClick here for file"} +{"text": "Regadenoson is a new vasodilator myocardial stress agent that is easier-to-use and more tolerable than adenosine. We demonstrate that, in patients undergoing cardiovascular magnetic resonance myocardial perfusion imaging, regadenoson is safe and effective in producing hyperemia and identifying the need for future revascularization.Regadenoson is a new vasodilator myocardial stress agent that selectively activates the A2A receptor. Unlike adenosine, regadenoson is easier to administer and results in fewer side effects. Although extensively studied in patients undergoing nuclear myocardial perfusion imaging (MPI), its performance in cardiovascular magnetic resonance (CMR) MPI remains unknown. The aim of this study was to assess the safety and tolerability of regadenoson and determine its ability to produce hyperemia and predict subsequent coronary revascularization in patients undergoing CMR-MPI.120 patients were prospectively enrolled to receive CMR-MPI with regadenoson. Patients with contraindications to CMR-MPI or regadenoson were excluded. Short-axis slices were obtained at three levels of the left ventricle (LV) during first pass of Gadolinium-DTPA(0.075 mmol/kg at 4 ml/sec) for 50 consecutive heart beats. Images were acquired using a hybrid gradient echo/echo planar imaging sequence. Imaging was performed 1 minute after injection of regadenoson (0.4mg) and then repeated 15 minutes after injection of aminophylline (125mg) under resting conditions. Perfusion defects were defined as subendocardial hypointensity in a coronary distribution at stress, involving \u226525% wall thickness, and persisting for \u22652 heart beats following peak enhancement of the LV cavity. In a subgroup of patients (n=99), custom software was used to generate time intensity curves and to compare the myocardial upslope of the midventricular slice during stress and rest. All subjects were followed for 3 months for the occurrence of coronary revascularization.Overall, 51/120 (43%) of patients were female with an average age of 55\u00b115 years and body mass index of 29\u00b16 kg/m2. Baseline patient characteristics include: coronary artery disease (33%), diabetes (38%), hypertension (56%), and hypercholesterolemia (95%). The average resting blood pressure and heart rate were 124/61mmHg and 70bpm, respectively. Peak heart rate after regadenoson administration was 98bpm (p<0.001). Most patients (87%) experienced side effects from regadenoson including shortness of breath (34%), flushing (23%), and chest discomfort (17%). No EKG changes or residual side effects persisted in any patient at completion of study. The average myocardial upslope increased significantly between rest and stress conditions , reflecting the expected hyperemic effect of regadenoson. Perfusion defects were visually apparent in 33/120 (28%) patients. Revascularization occurred in 8/120 (7%) patients (Figure Regadenoson is safe, easy-to-use, and effective in producing hyperemia and identifying need for future revascularization in patients undergoing CMR-MPI.Research was funded in part by a research grant from Astellas Pharmaceutical."} +{"text": "SPHKAP and DPP6) with significantly methylated promoters were of interest and further analysis of their expression showed them to be repressed in AML. We also demonstrated considerable cytogenetic subtype specificity in the methylomes affecting different genomic features. Significantly distinct patterns of hypomethylation of certain interspersed repeat elements were associated with cytogenetic subtypes. The methylation patterns of members of the SINE family tightly clustered all leukemic patients with an enrichment of Alu repeats with a high CpG density (P<0.0001). We were able to demonstrate significant inverse correlation between intragenic interspersed repeat sequence methylation and gene expression with SINEs showing the strongest inverse correlation (R2\u200a=\u200a0.7). We conclude that the alterations in DNA methylation that accompany the development of AML affect not only the promoters, but also the non-promoter genomic features, with significant demethylation of certain interspersed repeat DNA elements being associated with AML cytogenetic subtypes. MeDIP-seq data were validated using bisulfite pyrosequencing and the Infinium array.Methylated DNA immunoprecipitation followed by high-throughput sequencing (MeDIP-seq) has the potential to identify changes in DNA methylation important in cancer development. In order to understand the role of epigenetic modulation in the development of acute myeloid leukemia (AML) we have applied MeDIP-seq to the DNA of 12 AML patients and 4 normal bone marrows. This analysis revealed leukemia-associated differentially methylated regions that included gene promoters, gene bodies, CpG islands and CpG island shores. Two genes ( MLL gene is involved in many chromosomal translocations in leukemia EZH2 gene, which encodes a histone methyltransferase, is subject to inactivating monoallelic and biallelic mutations in myelodysplastic/myeloproliferative neoplasms DNMT3A, which encodes a DNA methyltransferase Acute myeloid leukemia (AML) is a heterogeneous form of cancer in which many molecular and cytogenetic somatically acquired events have been described MeDIP (methylated DNA immunoprecipitation) is one of the main approaches for detection of DNA methylation 8 reads were generated of which 53% could be mapped uniquely to the reference human genome (NCBI 36/hg18). The coverage of the 27 million CpG sites in the reference genome ranged from 63% to 87% for the 16 datasets (2\u200a=\u200a0.8) In order to perform a genome wide analysis of DNA methylation in AML, we applied the MeDIP-seq technique to DNA samples from 12 AML patients: 3 with t translocation, 3 with the t translocation, 3 with trisomy 8, and 3 with a normal karyotype (NK). We used 4 unrelated NBMs as control samples . A totaldatasets . Furtherdatasets [13]. Foctively) . These cFirstly, we analyzed AML and NBM whole genome data to assess the significance of differences in global DNA methylation. We categorized DNA methylation into 5 groups; <0.2, 0.2\u20130.4, 0.4\u20130.6, 0.6\u20130.8, 0.8\u20131.0 Batman scores ID4 has been shown to act as a putative tumour suppressor gene in AML DCC is hypermethylated in follicular lymphoma TERT increases the risk of familial AML DPP6 is down regulated in melanoma SPHKAP plays a role in the sphingosine phosphorylation pathway that induces tumor progression and invasion MYOD1, SOX14, FOXA2, FOXB2, RUNX1 and PAX1.Next, we investigated whether methylation of specific genomic features could discriminate between AML and NBM. Consequently, we searched for differentially methylated regions (DMRs) in 4 genomic categories; promoters, gene bodies, CGIs and CGI shores using empirical Bayes statistics (Bioconductor's Limma R package). This analysis indicated that 105 gene promoters and 704 CGIs showed significant differences in methylation between AML and NBM . 80% of MEIS1/2, TOP3B, CDH13, ST6GAL2 in t AML, DOK6, NCOR2 in t AML, ELK1, VMO1 in NK AML, SNX16, HHEX in trisomy 8 AML. Based on a two-dimensional cluster analysis AML. By contrast, trisomy 8 AML showed the lowest number of identified DMRs. Most of total DMRs in each AML subtype were hypermethylated except in trisomy 8 AML where only 40% of total identified DMRs were hypermethylated. For all AML subtypes, the hypermethylated DMRs was located mostly in CGIs where the preferential methylation was found in CGIs located outside the promoters . Additioanalysis and a paanalysis AML subtA considerable advantage of high-throughput sequencing is the ability to investigate repeated elements, which would cross-hybridize on a microarray chip DPP6, SPHKAP), 2 DMRs that were hypermethylated in 2 AML subtypes; ST6GAL2 in t, HHEX in trisomy 8 AML and an AluJb repeat that was differentially hypomethylated in t AML. The results revealed statistically significant differences in DNA methylation as was detected by MeDIP-seq .We sought to determine the relationship between the methylation profiles and gene expression in AML patients. Therefore, an array based gene expression profiling for 6 of the AML samples that had available RNA was performed. Correlating the gene expression to corresponding DNA methylation on an average scale revealed strong significant inverse correlation in promoters , CGIs and their parallel CGI shores Furthermore, we investigated the relationship between differential gene expression and DNA methylation by integrating our promoter MeDIP-seq data with published gene expression data DPP6 , SPHKAP and ID4 . The analysis was performed in 30 AML patients (including 4 AML patients who were previously involved in MeDIP-seq experiment), cancer cell lines and normal tissues using real time RT-PCR. The results showed that DPP6, SPHKAP and ID4 were down regulated in AML patients ] on the expression of both SPHKAP and DPP6 genes in 2 AML cell lines (OCI-AML2 and CTS). This confirmed that the demethylating treatment was able to restore the expression of both genes showed a significant methylated CGI located in its gene body (between exon 2 and exon 3) in trisomy 8 AML against the rest of the groups AML . Treating CTS and OCI-AML2 cell lines with DAC showed an increase in HHEX gene expression , we vali\u200a9\u00d710\u22126) . This de\u200a9\u00d710\u22126) . Next, ws P<0.5) . Correlapression . NotablyWe have used MeDIP-seq to establish the global methylome for AML. The comprehensive nature of this study (12 independent primary tumors) has allowed us to investigate the potential epigenomic role of not only gene promoters but also genomic elements, including CGIs and repeated elements. It is possible to draw some general conclusions from this study. Firstly, comparison of the whole genome methylation of the leukemia with the controls showed that leukemic DNA was only 2.7% less methylated. This contrasts with the generally held view that cancer is characterized by global hypomethylation amounting to a 10%\u201320% difference Previous genome wide epigenetic studies in leukemia have, for technical reasons, focused on gene promoters and/or CGIs SPHKAP) encoding sphingosine kinase anchoring protein in AML was of particular interest since SPHKAP was down regulated in both AML patients and cancer cell lines. The SPHKAP protein was identified through its interaction with and regulation of SPHK1 activity SPHKAP was recently identified as a member of A-kinase-anchoring proteins (AKAPs) SPHK1 catalyses the phosphorylation of sphingosine to sphingosine-1-phosphate (S1P) which promotes cell survival and proliferation SPHK1 is overexpressed in a range of cancers and has been proposed as a novel target for cancer therapeutics SPHKAP and resultant effects on the lipid signaling pathways remains unclear. However, it is of interest to note that expression of the RUNX1 gene, which is involved in the t translocation in AML, has recently been linked to regulation of key enzymes involved in sphingolipid metabolism Although our study did not include all possible subtypes of AML it did robustly identify specific methylation targets that were confirmed in a larger series of AML samples using pyrosequencing technology. Amongst the novel targets identified in this study the frequent methylation of the promoter of the gene (DPP6 gene. The DPP6 gene was reported as a hypomethylated gene in colon cancer DPP6 gene became hypermethylated in some tumors and hypomethylated in other types of tumors as the promoter hypomethylation is important in activation some oncogenes and in provoking loss of imprinting (LOI) The second novel promoter associated DMR, which appeared in our study to be down regulated in AML patients, was the Since up to 45% of the human genome consists of repetitive sequences which are not analyzed by array-based methods, it was desirable to extend the analysis to include such sequences. Our study suggests that both the hyper- and hypo-methylation of individual members of the LINE, SINE and LTR families can readily discriminate AML from NBM. Although we found some members of the satellite repeat family exhibiting differential methylation, the discrimination was much less clear. Differences between satellite methylation and interspersed repeat methylation were previously reported in leukemia and in bladder cancer The establishment of such high resolution AML methylomes not only reveals the subtle epigenetic changes involved in leukemogenesis but also has potentially important clinical implications. It has been shown that detection of methylated sequences in clinical remission for AML has the power to predict relapse risk for those patients We applied MeDIP-seq to 12 AML patients. Diagnosis of leukemia was based on clinical and morphological features http://maq.sf.net/ and Li et al.) and Bowtie Only high quality genomic DNA was subjected to MeDIP-seq protocol [25], [5Quantile normalization was performed to reduce the possible variations among the laboratory assays and to facilitate the comparison of the genes across all the samples. This method is based upon the concept of quantile-quantile plot extended to n dimensions (where n is the number of samples) DMR was defined as a differential methylated region with uncorrected P<0.05 and an absolute methylation difference >0.25 Batman score . This threshold was defined using the distribution of absolute differences in methylation (Batman) scores of all covered genomic features between normal and leukemic samples . Since tHierarchical clustering and pair-wise comparison were formed using Pearson correlation coefficient to construct the distance matrix among samples together with \u2018Ward\u2019 linkage clustering method For the comparison of the whole genome methylation between AML and NBM, we have categorized DNA methylation into 5 groups and used Fisher's exact test provided with R to investigate if there was a statistically significant difference in the global DNA methylation http://hgdownload.cse.ucsc.edu/goldenPath/hg18/database/).Repeated regions were obtained from nested RepeatsRM327 table downloaded from UCSC genome annotation database (http://qmua.cdb.riken.jp).500 ng genomic DNA was bisulfite converted using EZ DNA methylation \u2122 kit (Zymo research). PCR amplification of the bisulfite converted DNA was performed through 42 cycles at 55\u00b0C annealing temperature, the primers used for the PCR are provided in 500 ng genomic DNA was bisulfite converted using EZ-96 DNA kit TM (Zymo research) specified by the manufacturer. The converted DNA was PCR amplified using primers for each set of genes; the primers were designed using PyroMark Assay design 2.0 (primers provided in 500 ng genomic DNA was bisulfite converted and hybridized onto Illumina Infinium HumanMethylation 27 BeadArrays according to the manufacturer's protocol 6 thawed cells using Trizol (Gibco-BRL) purification method. The Applied Biosystems Human Genome Survey Microarray (P/N 4337467) was used for the expression profile http://www.R-project.org). The array expression data has been deposited in a MIAME compliant database GEO accession number: GSE34722, .Good quality RNA was available for 6 AML patients involved in MeDIP-seq experiment. Total RNA of AML patient samples was extracted from a total of 10\u201320\u00d710SPHKAP, DPP6 and ID4. Normal lymph node was run on every plate to ensure the consistency across the runs. 18s RNA acquired from ABI was used as the endogenous control. All primers were designed and generated by ABI.We carried out RT-PCR starting with total RNA (1 \u00b5g) for cDNA synthesis using random primer with Superscript III reverse transcriptase performed as per the manufacturer's instructions (Invitrogen). Maximum cDNA concentration was used in a volume of 10 \u00b5l per run. Real time was carried out using Taqman universal master mix II as per the manufacturer's instructions 6 AML cell line cells were maintained in culture containing RPMI-1640 with 10% FBS. Cells were treated by DAC through adding freshly prepared 5 \u00b5M DAC in dimethylsulfoxide (DMSO) to the standard media. DAC treatment was repeated at 48 hours. Control cell lines were treated identically except that they were treated with standard media to which DMSO only had been added. We assayed the cells for gene expression 72 hours after the treatment.Twenty-four hours before adding 5-aza-2\u2032 deoxycytidine (DAC) (Sigma), 1\u00d710Text S1Supporting method MeDIP-seq protocol.(DOC)Click here for additional data file.Figure S1Saturation analysis of MeDIP-seq samples The saturation analysis investigates whether the number of unique reads is sufficient to generate a saturated and reproducible methylation profile of the reference genome. The higher Pearson correlation r the greater assurance of the reproducibility of the methylation profiles. Sample study number is identified in (DOC)Click here for additional data file.Figure S2Correlation between MeDIP-seq and Illumina Infinium array. A significant positive correlation was found between MeDIP-seq and Illumina Infinium array in the three MeDIP-seq samples.(DOC)Click here for additional data file.Figure S3Pair-wise comparison between AML and NBMs in 4 genomic regions. Red colored spots indicate high similarity and white colored spots low similarity. CGIs (C) showed the highest similarities between AML subtypes and a clear discrimination from NBMs. (A) Promoters, (B) Gene bodies, (D) CGI shores.(DOC)Click here for additional data file.Figure S4Estimating the number of clusters in data set consisting of differentially methylated CGIs using prediction strength method The consecutive number of clusters is given on the x-axis. The vertical bars illustrate the standard error of the prediction strength over 5 cross-validation folds. Prediction strength above 0.8 indicates well-separated clusters. Dividing the CGI associated DMRs data set into two clusters, referring to AML and NBM groups, gives the highest stability.(DOC)Click here for additional data file.Figure S5Characters of DMRs in AML subtypes. Venn diagrams showed few overlapped DMRs between AML subtypes with no common DMRs detected between the all 4 AML subtypes.(DOC)Click here for additional data file.Figure S6Pair-wise comparison between AML subtypes in 4 genomic regions. t AML subtype is discriminated distantly from all the other AML subtypes. (A) Promoter, (B) gene bodies, (C) CGIs, (D) CGI shores.(DOC)Click here for additional data file.Figure S7Pair-wise comparison between AML and NBMs in repeat sequences. (A) SINEs showed the highest similarities between AML subtypes among other repeats; (B) LINEs and (C) LTRs.(DOC)Click here for additional data file.Figure S8Pair-wise comparison between AML subtypes in repeat sequences. There was clear discrimination between AML subtypes in (A) SINEs, (B) LINEs and (C) LTRs.(DOC)Click here for additional data file.Figure S9Direct bisulfite sequencing of significant differentially methylated genes/genomic regions in MeDIP-seq samples. For all figures, the horizontal line represents the position of each CpG investigated and the vertical line is the percentage of the methylation at particular CpG site from 0\u2013100%. The analysis was performed using QUMA.(DOC)Click here for additional data file.Figure S10Pyrosequencing results of candidate genomic regions in AML patients, AML cell lines and NBMs. (a) SPHKAP, (b) DPP6, (c) ST6GAL2, (d) HHEX and (e) Alu repeat. N refers to the number of samples tested for each investigated genomic region. Kruskal-Wallis test showed significant methylation difference among the groups (P<0.0001) for all tested genes and repeat. Dunn's multiple comparison tests showed that there was significant methylation difference between AML patients and NBMs in SPHKAP and DPP6 (P<0.05). Also, there was significant methylation difference between AML samples and AML cell lines in all investigated genes (P<0.05) except in the repeat.(DOC)Click here for additional data file.Figure S11HHEX gene methylation and expression assay. (a) HHEX gene methylation (a significant methylated CGI located within the body of HHEX gene) among different AML patients. (b) HHEX gene expression among different AML patients. (c) Effect of DAC on HHEX expression in AML cell lines. HHEX gene expression was measured relative to NBM, PB\u200a=\u200aperipheral blood from healthy donors.(DOC)Click here for additional data file.Figure S12Histograms of uncorrected P values after testing the equality of the methylation means between groups. (a) in 4 genomic regions and (b) in repeats. When investigating the data with equal means between groups, the P values were expected to be uniformly distributed across the unit interval (blue line). Comparing the distribution of uncorrected P values to the uniform distribution expected for random data revealed enrichment of P value<0.05 (red line) indicating differential methylation pattern. Satellites did not show a specific distribution of uncorrected P values across the samples. High frequencies of P values<0.05 across the samples were observed in the other tested repeats; SINEs, LINEs and LTRs.(DOC)Click here for additional data file.Figure S13Histogram illustrating the distribution of uncorrected P values after testing equality of methylation between normal and leukemic samples for all genomic features and repeats. For random data the distribution is expected to be uniformly distributed across the unit interval . The frequency of P values<0.05 (red line) is higher than expected with particular enrichment of P values<0.001.(DOC)Click here for additional data file.Table S1Patients and control samples.(DOC)Click here for additional data file.Table S2Criteria of reads generated from Illumina GAII,(DOC)Click here for additional data file.Table S3Description of the genomic regions from MeDIP-seq results.(DOC)Click here for additional data file.Table S4DMRs identified in AML versus NBM in 4 genomic regions.(DOC)Click here for additional data file.Table S5DMRs identified in AML subtypes in 4 genomic regions.(DOC)Click here for additional data file.Table S6DMRs associated with repeats. AML versus NBM, (c) between AML subtypes.(DOC)Click here for additional data file.Table S7Direct bisulfite sequencing validation of selected genomic regions.(DOC)Click here for additional data file.Table S8Pyrosequencing validation of selected genomic regions. (a) AML versus NBM, (b) in AML subtypes.(DOC)Click here for additional data file.Table S9DNA methylation of over- and under expressed genes in t, t & NK AML subgroups that were included in MeDIP-seq experiment.(DOC)Click here for additional data file.Table S10DNA methylation of over- and under expressed genes in trisomy 8 AML subgroup that was included in MeDIP-seq experiment.(DOC)Click here for additional data file.Table S11The overlapped genes between the results of MeDIP-seq study and array-based study for t AML.(DOC)Click here for additional data file.Table S12The overlapped genes between the results of MeDIP-seq study and array-based study for t AML.(DOC)Click here for additional data file.Table S13Direct bisulfite sequencing primers.(DOC)Click here for additional data file.Table S14Pyrosequencing primers.(DOC)Click here for additional data file.Dataset S1Differentially methylated promoters.(XLS)Click here for additional data file.Dataset S2Differentially methylated gene bodies.(XLS)Click here for additional data file.Dataset S3Differentially methylated CGIs.(XLS)Click here for additional data file.Dataset S4Differentially methylated CGI shores.(XLS)Click here for additional data file.Dataset S5Differentially methylated SINEs.(XLS)Click here for additional data file.Dataset S6Differentially methylated LINEs.(XLS)Click here for additional data file.Dataset S7Differentially methylated LTRs.(XLS)Click here for additional data file."} +{"text": "Sepsis and septic shock lead to the multiorgan damage by extensive release of inflammatory mediators. Regenerative mechanisms include such regimens as stem cells which differentiate towards specific tissues. Also, in the course of the systemic inflammation the disruption of various regulatory axes occurs, including chemokines and complement proteins . Among other functions these axes maintain stem cell circulation and recruitment . The aimBlood samples were obtained from five patients with sepsis or septic shock on the second day after diagnosis. Blood from five healthy volunteers served as control. Samples were stained with the panel of antibodies against: CD45, lineage markers (Lin), CD34, CD133, VEGFR2 and isotypic controls. Cells were analyzed by flow cytometry and the total cell count per milliliter was calculated.+CD133+CD45+Lin-, endothelial progenitor cells (EPCs) CD34+CD133+VEGFR2+; and primitive nonhematopoietic stem cells. In the blood of septic patients we found: HSCs (5/5), median level 96/ml; EPCs (5/5), median 48/ml; and nonhematopoietic stem cells (4/5), median 48/ml. Whereas in the control group the results were as follow: HSCs (5/5), median level 644/ml; EPCs (5/5), median 70/ml; and nonhematopoietic stem cells (0/5). Two of five patients died of septic shock. A trend to lower number of HSCs in nonsurvivors was observed.On the basis of cell surface phenotype the following stem cell subpopulations were distinguished: hematopoietic stem cells (HSCs) CD34Stem cells can be identified phenotypically in the blood of septic patients and healthy volunteers. However, the circulating primitive nonhematopoietic stem cells could not be detected under physiological conditions. Furthermore, we suggest that stem cells analysis may have serve as prognostic tool in the future."} +{"text": "To evaluate the role of imaging in aiding surgical management and assess whether imaging resulted in unnecessary mastectomies.A retrospective analysis of 165 patients with invasive breast cancer treated with neoadjuvant chemotherapy (NAC) (2005 to 2009). Bilateral cancers were analysed separately . Data collected include tumour extent on mammogram and MRI, radiology advice for surgery, surgery performed, and whole tumour size on postsurgical specimens. MRI report recommended wide-local excision (WLE), mastectomy, or WLE with multidisciplinary team discussion (MDTD) based on imaging. We used residual tumour size <40 mm on pathology as a cut-off for suitability of WLE.Radiology advice regarding surgery was suggested in 167 cases and advice was followed in 142 (85%) cases. Fifty-nine cases were suggested for WLE; 46 (78%) underwent WLE with six requiring further surgery (no further disease in four) and 13 (22%) underwent mastectomy based on patient's choice or clinical decision. Ninety-one cases were suggested for mastectomy; two (2%) had successful WLE due to patient's choice (one had therapeutic mammoplasty) and 89 (98%) had mastectomy. Seventy-four were appropriate and based on pathology 15 may have had inappropriate mastectomies as residual disease <40 mm. Seventeen cases were considered for WLE with MDTD, seven (41%) had successful WLE and 10 (59%) had mastectomy; all were appropriately advised when compared with final pathology.This series suggests that NAC offers more opportunities for breast conservation than are being realised. Multifocal disease and extensive calcifications may not mandate mastectomy if the patient responds well to chemotherapy."} +{"text": "Our virtual screening protocol involves the fast docking of a small molecule library to rigid protein followed by redocking of top hits using a method that incorporates both ligand and protein flexibility. Subsequently, the top ranking compounds were prioritized using molecular dynamics simulation based binding free energy calculation. The result of biological assay and subsequent similarity search resulted in the identification of two classes of small molecules that shared biaryl urea scaffold. Both of these chemical classes displayed moderate inhibitory potency against SUMO E1. The most potent compound of each class inhibited the in vitro sumoylation with an IC50 of 11.1 and 13.4 \u03bcM. These compounds inhibit sumoylation by blocking the formation of SUMO-E1 thioester intermediate. Our study presents starting points for the development of novel therapeutic agents against various diseases targeting SUMO E1.Sumoylation is a post-translational modification affecting diverse cellular processes including DNA replication and repair, chromosome packing and dynamics, genome integrity, nuclear transport, signal transduction and cell proliferation . Sumoyla"} +{"text": "Knockout mice deficient in the gap junction gene connexin43 exhibit developmental anomalies associated with abnormal neural crest, primordial germ cell, and proepicardial cell migration. These migration defects are due to a loss of directional cell movement, and are associated with abnormal actin stress fiber organization and a loss of polarized cell morphology. To elucidate the mechanism by which Cx43 regulates cell polarity, we used a wound closure assays with mouse embryonic fibroblasts (MEFs) to examine polarized cell morphology and directional cell movement. Studies using embryonic fibroblasts from Cx43 knockout (Cx43KO) mice showed Cx43 deficiency caused cell polarity defects as characterized by a failure of the Golgi apparatus and the microtubule organizing center to reorient with the direction of wound closure. Actin stress fibers at the wound edge also failed to appropriately align, and stabilized microtubule (Glu-tubulin) levels were markedly reduced. Forced expression of Cx43 with deletion of its tubulin-binding domain (Cx43dT) in both wildtype MEFs and neural crest cell explants recapitulated the cell migration defects seen in Cx43KO cells. However, forced expression of Cx43 with point mutation causing gap junction channel closure had no effect on cell motility. TIRF imaging revealed increased microtubule instability in Cx43KO cells, and microtubule targeting of membrane localized Cx43 was reduced with expression of Cx43dT construct in wildtype cells. Together, these findings suggest the essential role of Cx43 gap junctions in development is mediated by regulation of the tubulin cytoskeleton and cell polarity by Cx43 via a nonchannel function. Gap junctions are specialized cell junctions that contain hydrophilic membrane channels that allow the passive diffusion of ions and small molecules between cells Connexin 43 knockout (KO) mice die shortly after birth from cardiac defects associated with pulmonary outflow obstruction and coronary anomalies In the Cx43 KO mouse, abnormal patterning of the coronary arteries are accompanied by outflow obstruction associated with conotruncal pouch tissue containing ectopic and disorganized deployment of endothelial and vascular smooth muscle cells While the importance of Cx43 in cell motility is well documented, the underlying mechanism remains unclear, and whether this requires the gap junction channel is unknown. Directional cell migration requires polarized alignment of the cytoskeleton with the microtubule organizing center (MTOC) and Golgi positioned forward facing at the cell's leading edge Cx43 interactions with the cytoskeleton have been suggested by several previous studies. The Cx43 carboxy terminus (CT) has been shown to bind ZO-1 To investigate whether modulation of cell polarity and polarized cell migration by Cx43 may involve Cx43 regulation of microtubule dynamics, we used a wound healing assay with mouse embryonic fibroblasts (MEFs) and NIH3T3 cells to quantitatively assess cell polarity and directional cell migration. Using this wound healing assay, we also examined the requirement for the previously identified Cx43 tubulin binding domain with forced expression of a Cx43 construct with the tubulin binding domain deleted (Cx43dT). To further assess the requirement for the gap junction channel in Cx43 modulation of cell motility, we examined the expression of a Cx43 mutation (Cx43Y17S) recovered from an oculodentodigital dysplasia (ODDD) patient. This Cx43 mutant protein was previously shown to have no gap junction channel activity, but nevertheless can make gap junction plaques at the cell surface To assess the role of Cx43 in directional cell migration, we generated primary mouse embryonic fibroblasts (MEFs) from Cx43 wildtype and KO mouse embryos for assessment of cell motility using a well described wound healing assay Cells undergoing directional cell locomotion exhibit a polarized cell morphology that is essential for productive cell migration. Typically the Golgi apparatus and microtubule-organizing center (MTOC) are situated in the forward facing direction or leading edge of the cell and the microtubule cytoskeleton is aligned with the direction of cell migration The perturbation of the microtubule cytoskeleton in the Cx43 KO MEFs is likely central to the cell migration defects observed with Cx43 loss, and in this regard, it is interesting to consider the potential role of a previously identified Cx43 tubulin binding domain To determine whether these Cx43 fusion protein constructs could function normally, we transfected these expression vectors into gap junction deficient N2A cells and dye coupling was assessed following iontophoretic injection of sulforhodamine 101, a low molecular weight fluorescent tracer that can pass through gap junction channels. As expected, dye injection into N2A cells expressing a GFP control plasmid showed no dye transfer from the site of injection . In contTo determine whether the Cx43dT protein missing the tubulin binding domain may affect cell motile behavior, we transfected NIH3T3 cells with the Cx43dT-GFP, Cx43FL-GFP, or Cx43-GFP constructs and examined cell motile behavior using the same wound closure assay. NIH3T3 cells expressing the Cx43dT-GFP protein displayed a reduced rate of wound closure when comTo examine the role of the gap junction channel in the modulation of cell polarity and directional cell migration we generated a GFP C-terminal tagged Cx43 construct with a point mutation (Cx43Y17S-GFP) that eliminates gap junction channel function without perturbing protein trafficking or assembly of gap junction plaques at the cell surface To assess whether the mutant Cx43Y17S-GFP protein may alter microtubule dynamics, we examined the abundance of stabilized microtubules with Glu-tubulin immunostaining in NIH3T3 cells transfected with the Cx43Y17S-GFP construct. Cells expressing the Cx43Y17S-GFP construct showed no change in the level of Glu-tubulin expression as compaTo determine if Cx43 is also required for the modulation of cell migration in neural crest cells, we explanted neural crest cells from the E8.5 postotic hindbrain neural tube, the region where cardiac crest cells emerge . ExplantTo delineate the role of Cx43 in the modulation of microtubule dynamics, we used TIRF microscopy to examine microtubule membrane targeting events in Cx43 KO and wildtype MEFs. Cells were transfected with a tubulin-GFP construct to allow direct visualization of microtubules . We quanTo examine whether modulation of tubulin dynamics by Cx43 may require the previously described Cx43 tubulin binding domain, we used two colour TIRF microscopy to directly visualize Cx43-tubulin interactions in the living cell. For these experiments, the Cx43dT-dsRED construct was transfected into NIH3T3 cells to allow cell surface trafficking of Cx43dT via hetero-oligomerization with endogenous wildtype Cx43. Control experiments with time-lapse sequences obtained by TIRF imaging of cells expressing wildtype Cx43FL-dsRED showed punctuate Cx43 red dots trafficking to the cell membrane along GFP labeled microtubules . This isUsing a wound closure assay together with cell motion analysis, we showed Cx43KO MEFs have defects in directional cell migration. While the speed and directionality of cell migration were both decreased, cell protrusive activity was increased. These paradoxical results are explained by defects in cell polarity and polarized cell movement required for directional cell movement. Thus Cx43KO MEFs failed to reorient their MTOC and Golgi with the direction of cell migration and this was associated with a reduction in stabilized microtubules. MTOC/Golgi reorientation and microtubule stabilization are essential for directional cell migration significantly reduces cell migration in a wound closure model, they appear as odds with other wound healing assays Our TIRF analysis suggests Cx43 plays an important role in regulating tubulin dynamics. Thus Cx43KO MEFs showed increased microtubule instability. These findings are consistent with previous studies showing cell surface localized Cx43 are targets for microtubule capture Wnt1 KO mice have normal cell motility, but are not gap junction communication competent even though they had Cx43 gap junction plaques at the cell surface Overall, these findings are consistent with our previous studies showing no correlation between gap junctional communication level and changes in cell motile behavior in various transgenic and KO mouse models Together, these findings suggest Cx43 modulation of directional cell migration involves the regulation of tubulin dynamics, while cell-cell communication mediated by the Cx43 gap junction channel may be dispensable. We propose Cx43 may have two distinct functions, one involving formation of gap junction channels to mediate direct cell-cell communication, and a separate role in modulating the cytoskeleton. The integration of these two distinct functions in one protein may be evolutionarily advantageous as it may allow efficient integration of extracellular signals with intra/intercellular cues. We note there are a plethora of studies showing interactions of Cx43 with proteins associated with the cytoskeleton, including studies showing Cx43 interactions with N-cadherin 5\u2032-CCCCACTCTCACCTATGTCTCC-3\u2032, IMR5: 5\u2032-ACTTTTGCCGCCTAGCTATCCC-3\u2032) and KO primer pair . MEFs were allowed to grow in standard DMEM supplemented with 10% FBS and 50 U/ug/ml penicillin-streptomycin .All experiments were conducted in accordance with an approved animal protocol of the National Heart Lung Blood Institute (Protocol Number H-0175). Mouse embryonic fibroblasts (MEF) were obtained from E13.5 day embryos generated from interbreeding heterozygous Cx43 +/\u2212 animals as previously described Confluent MEFs grown in 4-well chamber slides were serum starved for 48 hr before use. The monolayer was wounded using a scratch made with a 20 ul micropipette tip for the analysis of wound closure. For time-lapse imaging, MEFs were cultured on a heated stage (37\u00b0C) in L-15 medium and imaged using a Leica inverted microscope . Time-lapse images were taken every 20 minutes over 5\u20137 hours using 10x and/or 40x objectives using Openlab 3.1.7 software . Images were subsequently analyzed using DIAS cell tracking software to determine cell speeds, directionality, and membrane flows. Directionality was defined as the net displacement achieved divided by the total distance traveled, with a cell moving in a straight line having a directionality of one.For immunohistochemistry, cells were fixed with 4% paraformaldehyde in PBS for 10\u201315 min, then rinsed with PBS, permeabilized with PBST (0.15%\u20130.3 triton in PBS) for 10\u201320 minutes and blocked with 5% FBS in PBST for 1 hour. This is followed by incubation in primary antibodies in PBST (+5% FBS) overnight at 4\u00b0C, then 3 washes in PBST, and then incubation with secondary antibodies diluted in PBST for 30 minutes. After 3 washes with PBS, the cells were mounted in Vectashield with DAPI . Cells were imaged using a Leica DMIR fluorescent imaging microscope with 40x and 63x oil objectives and Orca-ER C4742-95 CCD camera . Primary antibodies used included anti-vinculin, \u03b1-tubulin, \u03b3-tubulin, GM130 , and Glu-tubulin (Chemicon). Secondary antibodies were from Jackson Laboratories . Orientation of the MTOC/Golgi was scored according to the methods of Gomes & Gundersen Mus musculus full-length Cx43 (amino acids 1\u2013382) was generated by PCR and the fragment was cloned into either pcDNA3.1/CT-GFP-TOPO vector or pDsRed-Monomer-N1 vector . Cx43Y17S and Cx43-\u0394234\u2013243 (Cx43dT) mutants were generated using QuikChange II Site-Directed Mutagenesis Kit according to manufacturer's protocol. The GFP-tagged Tubulin was kindly supplied by Dr Chloe Bulinski .To generate GFP or DsRed-tagged Cx43 expressing vectors, cDNA encoding Embryos used for neural tube explant cultures were harvested at E8.5 day as described previously Neural crest cell migration was subsequently recorded and analyzed 24 hours after transfection as previously described Gap junction communication competency of cells expressing various Cx43 constructs was determined using Neuro-2a (N2A) cells . N2A cells where cultured on fibronectin coated 12 mm glass coverslips and transfected with plasmid DNA using PolyFect . 24\u201348 hours after transfection coverslips were placed on a heated stage of a Leica DMLFSA microscope in a dish containing prewarmed L-15 medium (+10% FBS) and dye-coupling was quantified by iontophoretic dye injection (0.5 nA current pulses at 1 Hz) of sulforhodamine 101 dye . Fluorescent images were collected every 30 seconds over 4 minutes using Cy3 filters. Dye-coupling was quantified by counting the percentage of dye containing transfected cells surrounding the injected cell.NIH3T3 or MEF cells were plated on fibronectin coated glass bottomed culture dishes and allowed to grow overnight. The next day cells were transfected with Cx43 and/or tubulin-GFP constructs using lipofectamine . Western blot anaylsis was used to confirm expression levels were comparable between groups . Cells wData is presented as mean \u00b1 SEM as analyzed using Instat 3 with one-way analysis of variance (ANOVA) and Bonferroni correction. P<0.05 was considered significant.Figure S1Relative expression levels of Cx43 isoforms following transfection into NIH3T3 cells. Separate Cx43 and EGFP western blots were run using rabbit polyclonal antibodies. Merge image was generated by lining up fragment ladders. * highlights location of GFP tagged Cx43 \u2020 highlights unbound GFP protein (GFP: 27 kb).(TIF)Click here for additional data file.Movie S1TIRF microscopy and time lapse imaging show microtubule polymerization/depolymerization events at the cell membrane of a Cx43 KO MEF. Images from this video sequence was used to generate the panels in (MOV)Click here for additional data file.Movie S2Time lapse imaging with single colour TIRF microscopy shows polymerization/depolymerization of a single microtubule, termed searching, at the cell membrane of a Cx43 KO MEF. Images from this video sequence was used to generate the panels in (MOV)Click here for additional data file.Movie S3Two colour TIRF microscopy and time lapse imaging showing a Cx43FL-DsRed plaque being transported to the cell membrane along a microtubule (from bottom to top of movie) in a NIH3T3 cell. Images from this video was used to generate the panels in (MOV)Click here for additional data file.Movie S4Two colour TIRF microscopy and time lapse imaging showing microtubule polymerization and targeting of Cx43FL-DsRed plaques in the cell membrane of a NIH3T3 cell. Images from this video was used to generate the panels in (MOV)Click here for additional data file."} +{"text": "To the Editor: Hepatitis E virus (HEV) is a positive-stranded, non-enveloped RNA virus of the family Hepeviridae that is considered to be the main causative agent of enterically transmitted acute hepatitis in abattoirs in Douala and Yaound\u00e9, Cameroon, and in slaughter slaps (areas) in Bamenda, Cameroon. Pigs were mainly of the local breed. In addition, pigs originating from extensive cross-breeding were sampled. Liver samples were collected during post-mortem inspection.,Viral RNA was extracted from liver samples by using the RTP DNA/RNA Virus Mini Kit II according to the manufacturer\u2019s instructions. Extracted RNA was analyzed for HEV RNA by using 2 nested reverse transcription PCRs (RT-PCRs) specific for open reading frame 1 (OFR 1) and ORF 2 of HEV and 1 sample from a male pig in Bamenda (1/39). All 167 samples from Douala were negative for HEV RNA. The sample from Bamenda showed a positive result for the nested RT-PCR specific for HEV ORF 1. Genetic distances calculated with partial nucleotide sequences of ORF 1 (280 nt) and ORF 2 (373 nt) between strain Yaounde56 and the most closely related HEV genotype 3 strains from Japan and Mongolia were 90% and 91%, respectively.At the amino acid level, the partial RNA-dependent RNA polymerase sequence (ORF 1) and the partial capsid protein sequence (ORF 2) of strain Yaounde56 were identical to those of HEV genotype 3 strains HEV/Gt3/HSD40/2009 (GenBank accession no. AFO71833) from Germany and swJ12\u20131 (GenBank accession no. BAC66273) from Japan. Thus, all mutations were silent.In agreement with distance analysis, phylogenetic reconstruction using partial nucleotide sequences of ORF 2 (278 nt) showed a close relationship of strains Yaounde56 and Yaounde94 with HEV genotype 3 strains . HoweverBecause the pig production cycle is shorter than that for cattle, pig production is a major economic activity in Cameroon. Most pigs in Cameroon are local raised, and extensive cross-breeding is used. The infection rate of pigs with HEV genotype 3 strains from Cameroon is lower than that of pigs from Europe. Thus, HEV genotype 3 seems to have an extensive distribution that includes Africa. Future studies should investigate how HEV genotype 3 strains contribute to sporadic HEV cases in Cameroon."} +{"text": "We report results of the genome-wide association studies for melanoma that were conducted by the GenoMEL Consortium. In the first study 2981 individuals with melanoma and 8408 study-specific control individuals of European ancestry were analysed, in the second experiment 2168 Australian individuals with melanoma and 4387 control individuals were genotyped for 317,000 or 610,000 single-nucleotide polymorphisms (SNPs). Three of the previously reported melanoma susceptibility loci reached genome-wide significance in both studies. Three new loci were found to be associated with melanoma risk in European GWAS: rs1801516 in ATM, rs45430 in MX2, rs13016963 in CASP8. A fourth locus near CCND1 remains of potential interest. An Australian GWAS identified a new susceptibility locus at 1q21.3 (rs 7412746) and potentially 1q42.12 (rs3219090). Further studies will be required to determine which gene or genes at the reported loci mediate melanoma risk."} +{"text": "We performed transcriptional profiling of mouse iPSC lines before and after excision of a polycistronic lentiviral reprogramming vector to systematically define the overall impact of persistent transgene expression on the molecular features of iPSCs. We demonstrate that residual expression of the Yamanaka factors prevents iPSCs from acquiring the transcriptional program exhibited by embryonic stem cells (ESCs) and that the expression profiles of iPSCs generated with and without c-Myc are indistinguishable. After vector excision, we find 36% of iPSC clones show normal methylation of the Gtl2 region, an imprinted locus that marks ESC-equivalent iPSC lines. Furthermore, we show that the reprogramming factor Klf4 binds to the promoter region of Gtl2. Regardless of Gtl2 methylation status, we find similar endodermal and hepatocyte differentiation potential comparing syngeneic Gtl2ON vs Gtl2OFF iPSC clones. Our findings provide new insights into the reprogramming process and emphasize the importance of generating iPSCs free of any residual transgene expression.Delivery of the transcription factors Oct4, Klf4, Sox2 and c-Myc via integrating viral vectors has been widely employed to generate induced pluripotent stem cell (iPSC) lines from both normal and disease-specific somatic tissues, providing an invaluable resource for medical research and drug development. Residual reprogramming transgene expression from integrated viruses nevertheless alters the biological properties of iPSCs and has been associated with a reduced developmental competence both The discovery that differentiated adult cells can be reprogrammed to a state of pluripotency through the introduction of a defined set of transcriptional regulators has opened new avenues for understanding and treating degenerative diseases in vivo and in vitroReprogramming was originally achieved via retroviral transfer of Oct4, Klf4, Sox2 and c-Myc (OKSM) Dlk1-Dio3 gene cluster, seems sufficient to predict the developmental potential of mouse iPSCs Gtl2, a member of the Dlk1-Dio3 cluster normally expressed from the maternally inherited allele, contribute poorly to chimeras and fail to produce viable mice through tetraploid complementation. To date, it has not been clear whether aberrant Gtl2 silencing in iPSCs results from the selection of a subset of previously mis-imprinted parental fibroblasts or occurs at some point during the reprogramming process. Understanding which of these differences are introduced during reprogramming and whether they are functionally relevant is critical since they may influence potential downstream therapeutic applications.More recent reports have raised additional controversies regarding subtle genetic and epigenetic differences between iPSCs and ESCs that arise during reprogramming We previously described the development of the STEMCCA (\u201cSTEM Cell CAssette\u201d) polycistronic lentiviral vector for the efficient generation of iPSCs Sox2-GFP knock-in postnatal mouse fibroblasts using either the EF1\u03b1-STEMCCA-RedLight-loxP vector (N\u200a=\u200a5) or the EF1\u03b1-STEMCCA-loxP vector (N\u200a=\u200a5). These vectors allow for the co-expression of three (OKS) or four (OKSM) factors respectively, as previously described To gain insight into the transcriptome changes that result from the removal of exogenous reprogramming factors we performed genome-wide gene expression analysis on iPSC lines before and after Cre recombinase-mediated deletion of the STEMCCA polycistronic vector. Ten iPSC clones were derived from Sox2-GFP ESC line are shown in The different groups of iPSC clones that were subjected to microarray analysis along with five ESC subclones obtained from a We performed a two-way ANOVA of the 25 samples to determine: a) the effect of transgene removal on the global transcriptome of iPSCs (Cre effect), and b) differences in gene expression between ESCs, 3 factor iPSCs and 4 factor iPSCs (cell effect). Comparing the datasets of pre-Cre and post-Cre iPSCs we found 2,327 significantly differentially expressed probesets . Some of these regulators participate in chromatin remodeling complexes that are required for the establishment and maintenance of the pluripotent state in vivo developmental competence of transgene-carrying iPSCs as well as their poor response to Activin A stimulation in vitroTo investigate the molecular changes brought about by the excision of STEMCCA the list of differentially expressed genes was uploaded into the online functional annotation tool DAVID. Gene ontology analysis indicated a number of significantly enriched GO terms that corresponded to metabolic and biosynthetic processes, tissue development, and morphogenesis . Most imRecent data indicates that the transcription factor c-Myc acts mainly during the early stages of reprogramming by inducing cell cycle changes consistent with self-renewal and/or promoting dedifferentiation Dlk1-Dio3 region on mouse chromosome 12qF1 is actively transcribed in fully pluripotent iPSCs but silenced in iPSC clones that lack the capacity to support the development of \u201call-iPSC mice\u201d Gtl2, a member of this cluster that is active in ESCs, was found aberrantly silenced in most iPSC clones despite being normally imprinted (\u223c50% CpG methylation) in the starting fibroblast population Recent studies have reported that the conserved imprinted Dlk1-Dio3 domain during reprogramming we first analyzed the expression values of Gtl2 in our microarray datasets but were unable to detect statistically significant differences in expression levels between ESCs and iPSCs by ANOVA (data not shown). Therefore we determined Gtl2 mRNA expression levels in the 20 iPSC clones via qRT-PCR. We identified two of 20 iPSC clones (5-Cre and 15-Cre) with low Gtl2 expression levels compared to control ESCs, suggesting these clones were \u201cGtl2OFF clones\u201d, possibly due to aberrant silencing of the imprinted Dlk1-Dio3 locus, as has been previously reported ON clones\u201d data Gtl2 transcription start site (TSS), but more importantly, a Klf4-binding site was identified within the Gtl2 imprinted domain next to the TSS (Gtl2 we performed chromatin immunoprecipitation (ChIP) followed by qPCR (ChIP-qPCR) analysis using an antibody directed against Klf4. The results, shown in Gtl2 promoter in ESC/iPSCs. Moreover, sustained ectopic expression of Klf4 in transgene-carrying iPSCs resulted in increased binding compared to transgene-free iPSCs, which showed values similar to ESCs persistent transgene expression prevents iPSCs from attaining an ESC-like transcriptional program; ii) iPSCs reprogrammed with and without c-Myc exhibit highly similar gene-expression profiles; iii) reprogramming results in aberrant imprinting of the Dlk1-Dio3 locus in some but not all iPSC clones generated with our polycistronic cassette; and iv) Klf4 binds strongly to the Gtl2 promoter and shows decrease binding upon removal of the reprogramming transgenes.Nanog and Oct4Lentiviral vectors seem to resist methylation-dependent silencing in mouse pluripotent cells in vitro cardiogenic potential in another study in vivo development, the reasons for these differences remain unclear. However, because c-Myc might participate in chromatin remodeling during reprogramming Pluripotency can be induced in the absence of c-Myc overexpression, albeit at a low efficiency and delayed kinetics Gtl2 locus during reprogramming seems to be important for the generation of iPSCs with full developmental potential OFF iPSC clones, while the same experimental approach yielded mostly clones with low to normal Gtl2 methylation levels when C57BL/6\u00d7129/sv fibroblasts were transduced. Moreover, two clones underwent Gtl2 silencing after transgene excision, suggesting that the reprogramming factors may be directly involved in this process. In support of this idea, binding sites for Oct4 and Klf4 were identified in the Dlk1-Dio3 region, and Chip-qPCR analysis revealed increased recruitment of Klf4 near the TSS of Gtl2 in transgene-carrying iPSCs. Our findings imply an active role of Klf4 (and possibly Oct4) in establishing the methylation status of Gtl2 and suggest that, when present at supraphysiological levels, Klf4 may protect this region from cytosine methylation through a mechanism similar to that described for the imprinted Igf2 gene Gtl2/Dlk1 DMRs in vivoGtl2 as recently shown Epigenetic modification of the In summary, we demonstrate that residual expression of exogenous reprogramming factors has a pervasive effect on the transcriptional program of mouse iPSCs and may also influence epigenetic signatures associated with full pluripotency. Although retroviral and lentiviral vectors undergo silencing in human pluripotent cells, our findings suggest that variegation effects as well as potential reactivation of the transgenes during differentiation could have a negative impact on the biological properties of iPSCs. Indeed, even small variations in the levels of pluripotency factors appear to have a profound effect on the early cell fate choices of ESCs Sox2-GFP/M2rtTA mice (mixed genetic background C57BL/6\u00d7129/sv) or M2rtTA mice (genetic background C57BL/6) and infected at passage 3. All mice employed in this study were male. All animal studies were approved by the Boston University IACUC committee. iPSC clones were isolated, expanded and characterized by immunofluorescence, alkaline phosphatase staining, teratoma formation and karyotyping as previously described Reprogramming of mouse fibroblasts with the EF1\u03b1-STEMCCA-loxP and EF1\u03b1-STEMCCA-RedLight-loxP lentiviral vectors was done essentially as described in Sox2-GFP/M2rtTA ESC line \u22121 leukemia inhibitory factor . Undifferentiated ESC/iPSCs were harvested by trypsinization and plated twice onto cell culture dishes to deplete the feeder cells before RNA isolation. Total RNA was isolated with the miRNeasy Mini Kit (Qiagen) and treated with RNase-Free DNase I (Qiagen) before performing gene expression analysis.The Gtl2, Rian and Dlk1 expression was carried out with Power SYBR Green Master Mix (Applied Biosystems). Primer sequences are described in One microgram of RNA was reverse-transcribed using the TaqMan Reverse Transcription Reagents kit (Applied Biosystems) according to the manufacturer\u2019s instructions. The STEMCCA and STEMCCA-RedLight polycistronic viral transcripts were amplified in a StepOnePlus real-time PCR system (Applied Biosystems) using Custom TaqMan\u00ae Expression Assays as described by the manufacturer. Endoderm induction and liver specification from iPSCs were evaluated using the following Taqman inventoried primers and probes (Applied Biosystems): Sox17 (Mm00488363_m1), AFP (Mm00431715_m1) and Alb1 (Mm00802090_m1). qRT-PCR analysis of http://david.abcc.ncifcrf.gov) was used to identify Gene Ontology (GO) terms and KEGG pathways that were enriched in the list of differentially expressed genes.The Mouse Gene 1.0 ST array (Affymetrix) was used for mRNA expression profiling following the manufacturer\u2019s protocol. Twenty-five raw data files obtained by the Affymetrix scanner passed data quality control steps prior to RMA (robust multiarray average) normalization using the Affymetrix Expression Console software. To determine the differentially expressed genes affected by either cell type (ESC/iPSC) or transgene excision (before Cre/after Cre), a two-way ANOVA method was applied to analyze all 25 samples comprised of 5 groups representing the differing cell types (first ANOVA variable) and the effect of transgene excision (second ANOVA variable). The differentially expressed genes were subsequently subjected to the multiple hypothesis test by using FDR adjustment. We have developed similar ANOVA methods for microarray analyses of iPSCs detailed previously Gtl2 IG-DMR and the ADS1341 assay for Gtl2 DMR.Genomic DNA was purified with the DNeasy Blood & Tissue Kit (Qiagen) and modified with sodium bisulfite using the EpiTect Bisulfite Kit (Qiagen) according to the manufacturer\u2019s instructions. Pyrosequencing reactions were conducted by EpigenDx using the ADS935 assay for ChIP was performed as previously described \u22124 M MTG, 50 ng/ml Activin A, 50 ng/ml BMP-4, 10 ng/ml bFGF, and 10 ng/ml VEGF . At day 5, EBs were trypsinized, plated on gelatin-coated plates and cultured in the same medium without Activin A but supplemented with 20 ng/ml EGF, 20 ng/ml HGF, 20 ng/ml TGF-\u03b1 , and 10\u22127 M dexamethasone.All ESC/iPSC lines were adapted to serum-free maintenance media Figure S1Characterization of iPSCs generated with the STEMCCA vectors. (A) Southern blot analysis was performed to select iPSC clones carrying a single copy of the polycistronic vector that is excised after treatment with Cre-recombinase. gDNA was digested with BamHI and probed using standard methods. Clones 5, 15, B and D are shown as an example. Each band represents a single viral integration that is not detected after exposure to Cre-recombinase. (B) iPSCs derived using the STEMCCA vectors display ESC-like colony morphology (Phase), Sox2-GFP reporter gene expression, and alkaline phosphatase activity (Alk Phos), and form teratomas containing tissues derived from all three germ layers after injection into immunocompromised mice. (C) Representative images of DAPI-stained metaphase chromosomes from actively growing iPSCs displaying a normal karyotype (2n\u200a=\u200a40) both before and after transgene excision.(TIF)Click here for additional data file.Figure S2Venn diagrams illustrating common and unique differentially expressed probesets between the different iPSC \u201ccell types\u201d and the ESC control group (top) or between iPSCs generated with 3 and 4 factors (bottom). Tables show the numbers of differentially expressed probesets according to increasing p values.(TIF)Click here for additional data file.Figure S3RT-qPCR analysis shows total and endogenous levels of the reprogramming factors in the 25 samples profiled by microarray.(TIF)Click here for additional data file.Table S1Differentially expressed probesets in pre-Cre and post-Cre iPSCs .(XLSX)Click here for additional data file.Table S2Gene ontology analysis of differentially expressed genes.(XLSX)Click here for additional data file.Table S32 values of genes associated with enriched GO terms.Log(XLSX)Click here for additional data file.Table S4KEGG pathways identified by DAVID (EASE score <0.05).(XLSX)Click here for additional data file.Table S52 values of genes that map to the TGF-beta signaling pathway.Log(XLSX)Click here for additional data file.Table S6Summary of the iPSC/ESC lines used in this study.(XLSX)Click here for additional data file."} +{"text": "We report a unique case in a 17 year old male patient of Algerian origin with two rare genetic conditions with overlapping clinical symptoms. Familial Mediterranean fever (FMF) is an autosomal recessive disorder characterized by sporadic, paroxysmal attacks of fever and serositis. Hereditary coproporphyria (HCP) is one of the type of acute hepatic porphyria resulting in neurovisceral symptoms caused by deficient activity of mitochondrial enzyme coproporphyrinogen oxidase. Both are considered rare differential diagnosis for acute abdominal pain.17 year old boy of Algerian origin presented with long history of recurrent episodes of fever, abdominal pain since infancy. He experiencied 3-4 attacks per year each lasting typically for 2-3 days. There was no family history.Patient was referred simultaneously to immunology and metabolic medicine for further assessment. Differential diagnoses considered at the time included: periodic fever syndromes, hereditary angioedema, vasculitis and porphyria.9/l (ref 2.00-7.50).FBC results over the year showed intermittent leucocytosis during acute attacks with elevated C-reactive protein (CRP) and plasma viscosity (PV). Serum amyloid A (SAA) was not measured. Investigations during quiescent phase showed normal levels of SAA but slightly elevated CRP 13.4 mg/l (ref <10) and neutrophilia of 9.9 x10Genetic investigations for periodic fever syndromes confirmed two pathogenic MEFV gene mutation on sequencing Exon 2 and 10 at p. Glu148Gln), supporting diagnoses of FMF. Sequencing for MVK and TNFRSF1A gene were negative.Investigations undertaken by metabolic medicine specialists revealed urine coproporphyrin III at 42.75nmol/mmol cret (1.2-24.8) with porphrin/creat Ratio of 54.6 nmol/mmol (Ref <28). Faecal porphrin were 1639nmol/g dry weight (ref <130) with faecal coproporphyrin III: I ratio at 19.51 (Ref <2). The results confirmed diagnosis of HCP. Genetic tests are awaited for the patient. He has management plan for hereditary coproporphyria.Patient was commenced on colchicine at the dose of 500 micrograms twice daily. No further episodes of abdominal pain have been reported in the last 9 months since prophylaxis starting prophylaxis with colchicine.FMF and HCP are both recognised as rare causes of unexplained acute abdominal pain associated with fever. However there are several additional features which would favour FMF over HCP. The patient\u2019s ethic origin is more suggestive of FMF. The majority of HPC was reported in the North European ancestry. The onset of HCP also tends to be in puberty whilst FMF usually presents in childhood with initial attacks before the age of 10 in 65% of cases. Low grade persistent inflammatory response is a feature of FMF and not necessarily seen HCP. Finally apparent response to colchicine would further support diagnosis of FMF.None declared."} +{"text": "Cannabinoid receptors (CB) are G-protein coupled receptors involved in many physiological processes, like learning, appetite, nociception and others. Two subtypes termed CB1 and CB2) are involved in slightly different processes [ and CB2 By combining [2] 3D-QSAR, pharmacophore modeling, comparative modeling and molecular docking we could identify features responsible for receptor subtype specificity.Various pharmacophore models were derived from in-house libraries and data available in the literature. 3D structures of both receptor subtypes were created employing comparative modeling methods. The models were subjected to molecular simulations in solvated lipid bilayers to sample different receptor conformations. The models were used for molecular docking studies with small compound libraries. Employing the data obtained in the pharmacophore/3D-QSAR studies as additional constraints delivered valuable information on affinity and selectivity of the compounds towards CB1 and CB2. The results from this synergistic modeling approach could improve our understanding of the protein\u2013ligand interactions involved."} +{"text": "Under normal conditions, brain apolipoprotein E (apoE) is secreted and lipidated by astrocytes, then taken up by neurons via receptor mediated endocytosis. Free apoE is either degraded in intraneuronal lysosomal compartments or released. Here we identified a novel way by which apoE undergoes proteolysis in the extracellular space via a secreted neuronal protease. We show that apoE is cleaved in neuronal conditioned media by a secreted serine protease. This apoE cleavage was inhibited by PMSF and \u03b11-antichymotrypsin, but not neuroserpin-1 or inhibitors of thrombin and cathepsin G, supporting its identity as a chymotrypsin like protease. In addition, apoE incubation with purified chymotrypsin produced a similar pattern of apoE fragments. Analysis of apoE fragments by mass spectrometry showed cleavages occuring at the C-terminal side of apoE tryptophan residues, further supporting our identification of cleavage by chymotrypsin like protease. Hippocampal neurons were more efficient in mediating this apoE cleavage than cortical neurons. Proteolysis of apoE4 generated higher levels of low molecular weight fragments compared to apoE3. Primary glial cultures released an inhibitor of this proteolytic activity. Together, these studies reveal novel mechanism by which apoE can be regulated and therefore could be useful in designing apoE directed AD therapeutic approaches. Alzheimer's Disease (AD) is the most common cause of elderly dementia and is characterized by extracellular deposition of amyloid \u03b2-peptide (A\u03b2) plaques in brain The apolipoprotein E (apoE) protein is a 34-kDa glycoprotein associated with lipoproteins in periphery and brain. ApoE exists in humans as three major isoforms that differ from each other at amino acid positions 112 and 158 The level of APOE mRNA in brain is second only to liver. Astrocytes are the primary source of apoE in brain, although microglia and neurons have been shown to synthesize apoE under certain circumstances APOE genotype influences apoE protein levels in brain, with apoE4 levels lower than those of apoE3 Human recombinant (rec) neuroserpin, purified human plasma \u03b11-ACT, bovine pancreatic \u03b1-chymotrypsin, rec apoE3 and apoE4, purified cathepsin G from human leukocytes, purified thrombin from human plasma, and argotroban, were all obtained from Sigma. VLDL-apoE purified from human plasma was from rpeptide. Cathepsin G inhibitor was obtained from Calbiochem. PMSF and protease inhibitor mix were from ROCHE. D6E10 anti apoE antibody raised against apoE amino acids 141\u2013160 was from Cell Signaling.Rec human apoE3 and apoE4 were obtained by overexpression in Escherichia coli strain BL21-CodonPlus using pET32a vectors with thioredoxin as the fusion partner (Novagen). Vectors containing human cDNA for either apoE3 or apoE4 were kindly provided by Karl Weisgraber. The apoE-thioredoxin complex was obtained by sonicating the cells and removing debris by centrifugation at 32 000 rpm for 20 min using a TLA-100.4 rotor (Beckman). Thioredoxin was cleaved from the apoE with thrombin, and apoE was isolated by gel permeation chromatography on a column of Sephacryl S-300 (Amersham).\u22122). Cultures were grown in Neurobasal medium supplemented with B27 (Gibco). Primary glia were prepared from Sprauge Dawley rats at embryonic day 18\u201319 embryos. Briefly, brain tissue was homogenized sequentially through 19G, 21G and 23G needles in HBSS and cells were plated in DMEM containing 10% FBS. Medium was changed every second day. Animal handling and primary rat neuron and glia culture preparation were carried out in accordance with NIH guidelines for the care and use of laboratory animals, and were reviewed and approved by the Georgetown University Animal Care and Use Committee (protocol# 13-018-100076). Rats were euthanized humanely using CO2 chamber.Primary hippocampal and cortical neuronal cultures were prepared from Sprauge Dawley rats at embryonic day 18\u201319 embryos plated density (\u223c750 cells mm15% SDS-PAGE or NuPAGE (Invitrogen) gels followed by western immunoblotting were used for apoE proteolysis analysis.Excised pieces of silver stained gels were destained, dried, alkylated using 55 mM iodoacetamide and later rehydrated, followed by In-Gel digestion using 250 ng mass-spectrometry grade trypsin (Promega). Following digestion the reaction mixture was acidified to 1% tri-flouroacetic acid and dried to 5 ul volume which was loaded to a 2 cm\u00d775 um I.D. trap column of Easy nanoLC II HPLC (Thermo) for LC-MS/MS analysis. The LC was interfaced to a dual pressure linear ion trap mass spectrometer via nano-electrospray ionization. An electrospray voltage of 2.0 kV was applied to a pre-column tee. The mass spectrometer was programmed to acquire, by data-dependent acquisition, tandem mass spectra from the top 15 ions in the full scan from 400\u20131400 m/z. Dynamic exclusion was set to 30 s. Data processing and library searching was performed on Amazon Web Services-based cluster compute instances using the Proteome Cluster interface. All searches required semi-style tryptic cleavage, up to 1 missed cleavage, fixed modification of cysteine alkylation and variable modifications of methionine oxidation. XML output files were parsed and non-redundant protein sets determined using in-house scripts.Zymogram gels were obtained from Invitrogen and used according to manufacturer's instructions. Briefly, gels were run under non-reducing conditions and incubated in renaturing buffer to allow reactivation of enzymatic activity, followed by incubation in developing buffer for 24 h.Cell free conditioned medium was collected from DIV18-DIV22 primary rat hippocampal (HPC) or cortical (CTX) neurons. Glial serum-free conditioned medium was collected at DIV 14 when primary glial cells were washed twice with PBS, incubated with serum free DMEM for 4 h and then incubated with fresh DMEM for 24 h. 2 ug rec apoE was incubated with 100 ul final volume of either individual or mixed conditioned medium for the indicated time periods. All inhibitors used were diluted to 1\u00d7 concentrations from 100\u00d7 stocks.Human brain frontal cortex tissue was obtained from the Department of Pathology, Johns Hopkins University, Baltimore, MD . Tissue Immunoblot X-ray films were scanned and images were analyzed using ImageJ software. Statistical analysis of three independent experiments (n\u200a=\u200a3) was carried out using Student's t-tests. Asterisk (*) indicates significant p value in the range of 0.001 to 0.05. Error bars represent standard deviations.Previous studies suggested a critical role of a neuron specific protease in apoE cleavage The gradual shift towards lower molecular weight fragments as well their subsequent turnover with longer incubation periods in conditioned medium indicated stepwise proteolysis of apoE by hippocampal neurons. In order to test if a secreted enzyme activity was responsible for apoE proteolysis, we analyzed the metabolism of rec apoE in cell free conditioned medium collected from hippocampal neuronal cultures. As in the cellular assays, both apoE3 and apoE4 underwent stepwise proteolysis to generate similar fragments A. We caAlthough very efficient apoE proteolytic activity was present in hippocampal conditioned medium, the rate of apoE proteolysis varied greatly in conditioned medium obtained from primary neuronal cultures prepared at different times. This variation contributed to slightly different patterns of apoE proteolysis with respect to time in each experiment. Experiments which involved comparisons between apoE isoforms and proteolytic activities were performed using the same conditioned medium.To characterize the protease responsible for apoE proteolytic activity, we performed the apoE proteolysis in the presence of different protease inhibitors. Incubation of primary hippocampal neurons with 10 uM leupeptin or conditioned medium with 10 uM EDTA did not affect apoE proteolysis (data not shown). However, apoE proteolysis was significantly impaired by a complete protease inhibitor mix (Roche) and by the serine protease inhibitor PMSF A, B, suIn addition to inhibiting chymotrypsin and related proteases, PMSF also inhibits thrombin and cathepsin G. Thrombin inhibitor AGTB inhibited apoE cleavage by purified thrombin efficiently , while cNext, we carried out LC-MS analysis of apoE fragments. Rec apoE3 was incubated with hippocampal conditioned medium for 24 h and silver stained apoE3 fragment bands were cut from the gel, followed by trypsin digestion. Tandem MS identification of peptides confirmed the presence of apoE fragments generated by a chymotrypsin like protease . ControlIn order to asses the cellular specificity of the apoE degrading protease, we compared apoE proteolysis in conditioned medium obtained from hippocampal or cortical neurons and from mixed glia cultures. ApoE proteolysis was less efficient in cortical conditioned medium compared to hippocampal conditioned medium. Within 48 h, only about 50% of the initial apoE was metabolized by conditioned medium from cortical neurons A, 5B, wSERPINs (Serine protease inhibitors) are a family of predominantly secreted proteins which possess the ability to inhibit serine proteases. We hypothesized that a member of the SERPIN family such as \u03b11-antichymotrypsin1 \u03b11-ACT) or \u03b11-antitrypsin (AAT) might be responsible for inhibition of chymotrypsin-like protease in the above assay D. \u03b11-ACIn order to define the proteolytic activities released in neuronal conditioned medium, we performed casein zymography. In hippocampal conditioned medium we identified three major protease activities close to 25 kDa in size A. Pre-iUnlike in hippocampal conditioned medium, no proteolytic activities were detected in cortical and glia conditioned medium B. As exPrevious studies have addressed apoE proteolytic fragments in the presence of AD pathology in human brains Results presented here identify a role of a secreted neuronal serine protease in extracellular metabolism of apoE. Conditioned medium obtained from both hippocampal and cortical cultures mediates apoE proteolysis, generating a pattern of apoE fragments similar to that seen in live neuronal cultures. Less activity was observed from cortical neurons compared to hippocampal neurons. The apoE cleaveage sites and inhibition profile supports a role of chymotrypsin-like protease in extracellular apoE cleavage. We also found a crucial role of glia in regulation of apoE proteolysis.Several different enzymes, such as cathepsin D, thrombin, chymotrypsin-like serine protease and aspartic proteases, have been proposed to mediate apoE cleavage Earlier studies described identification and partial purification of chymotrypsin-like protease as an intraneuronal protease responsible for apoE cleavage Reduced apoE proteolysis in cortical conditioned medium might result from less expression or inefficient secretion of protease, but reduced apoE proteolysis in glial conditioned media results from the presence of serine protease inhibitor blocking apoE proteolysis. SERPINS such as \u03b11-ACT or AAT1 secreted by glia might be responsible for this effect. In support of this idea we detected active enzyme-inhibitor complexes between the hippocampal conditioned medium protease and the glia conditioned medium inhibitor that corresponded in size to a complex between \u03b11-ACT and purified \u03b1-chymotrypsin. Glia are responsible for most apoE biosynthesis and lipidation The abundance of SERPINs such as \u03b11-ACT and AAT1 might be responsible for the low levels of apoE fragments in healthy human and mice brain, CSF and plasma. Supporting this hypothesis, co-incubation of human plasma or CSF together with hippocampal neuronal conditioned medium did not induce apoE proteolysis (not shown), whereas incubation of VLDL-apoE purified from human plasma was cleaved efficiently . ConformThe presence of apoE fragments in AD brains within amyloid plaques or insoluble fractions has been reported ApoE directed therapeutics remain an important research area for treating most cases of AD Figure S1Metabolism of recombinant apoE by hippocampal neurons. Rec apoE4 protein (2 ug) was incubated with DIV22 primary hippocampal neurons for indicated time periods and full length apoE and its proteolytic products in conditioned medium as well as cellular lysates were analyzed by western immunoblotting. Cell pellet was lysed in 200 uls RIPA buffer and 20 ul lysate (1/10th) was loaded on gel, whereas 25 uls of total 1 ml (1/40 th) conditioned medium was loaded per sample. Time in hours, t (h) is indicated on top while a longer exposure of the same blot is shown in the right panel. . At t\u200a=\u200a0, in addition to full length apoE at 34 kDa, we also detected a \u223c25 kDa apoE band and low amounts of higher molecular weight aggregates. More than 50% of full length apoE was metabolized within 6 h of incubation on hippocampal neurons, with almost 80% degradation within 48 h.(JPG)Click here for additional data file.Figure S2Comassie staining of purified recombinant apoE. Purified rec apoE4 (rE4) protein (4 ug) and rec apoE3 (rE3) protein (0.7 ug) were loaded on SDS-PAGE and detected by comassie staining.(JPG)Click here for additional data file.Figure S3Stability of recombinant apoE in non-conditioned control culture medium. Commercially obtained rE3 and rE4 (2 ug each) were incubated with control hippocampal culture medium for indicated time periods followed by analysis using western immunoblotting. Time in hours, t(h) is indicated on top while a longer exposure of the same blot is shown in the right panel. ApoE levels remain unaltered during incubation and lower molecular weight apoE proteolytic fragments do not appear even after longer exposure.(JPG)Click here for additional data file.Figure S4Metabolism of human VLDL-apoE by secreted neuronal protease. Commercially obtained VLDL-apoE (2 ug) purified from human plasma was incubated for indicated times at 37\u00b0C with DIV22 conditioned hippocampal medium. ApoE proteolysis apoE was analyzed by western immunoblotting. Time in hours, t(h) is indicated on top.(JPG)Click here for additional data file.Figure S5Efficient inhibition of thrombin mediated apoE degradation by argotroban. rE3 and rE4 (2 ug each) were incubated with purified human thrombin (5 U) in presence and absence of 10 uM argotroban (AGTB) as indicated for 24 h followed by analysis using western immunoblotting. Time in hours, t(h) is indicated on top while a longer exposure of the same blot is shown in the right panel. Almost complete degradation of apoE occurred within 24 h by thrombin, with very efficient inhibition of both apoE3 and apoE4 degrdation by 10 uM argotroban.(JPG)Click here for additional data file.Figure S6Shorter exposures of apoE blots from .(JPG)Click here for additional data file.Table S1Table describes the human AD brains used for the study. BRC# represents the assigned number to each specimen by John Hopkins ADRC Brain Bank.(PDF)Click here for additional data file."} +{"text": "Sc negative. This feature will impact detection of Atypical/Nor98 scrapie cases in the field, and highlights the need to review current evaluations of the disease prevalence and potential transmissibility. Finally, an estimate is made of the infectivity loads accumulating in peripheral tissues in both Atypical/Nor98 and classical scrapie cases that currently enter the food chain. The results obtained indicate that dietary exposure risk to small ruminants TSE agents may be higher than commonly believed.Atypical/Nor98 scrapie was first identified in 1998 in Norway. It is now considered as a worldwide disease of small ruminants and currently represents a significant part of the detected transmissible spongiform encephalopathies (TSE) cases in Europe. Atypical/Nor98 scrapie cases were reported in ARR/ARR sheep, which are highly resistant to BSE and other small ruminants TSE agents. The biology and pathogenesis of the Atypical/Nor98 scrapie agent in its natural host is still poorly understood. However, based on the absence of detectable abnormal PrP in peripheral tissues of affected individuals, human and animal exposure risk to this specific TSE agent has been considered low. In this study we demonstrate that infectivity can accumulate, even if no abnormal PrP is detectable, in lymphoid tissues, nerves, and muscles from natural and/or experimental Atypical/Nor98 scrapie cases. Evidence is provided that, in comparison to other TSE agents, samples containing Atypical/Nor98 scrapie infectivity could remain PrP Following the bovine spongiform encephalopathy (BSE) crisis and the identification of its zoonotic properties, a sanitary policy has been implemented based on both eradication of transmissible spongiform encephalopathies (TSE) in food-producing animals and exclusion of known infectious materials from the food chain. Atypical/Nor98 scrapie is a prion disease of small ruminants identified worldwide. Currently it represents a significant part of the TSE cases detected in Europe. The restricted tissue distribution of Atypical/Nor98 scrapie agent in its natural host and the low detected prevalence of secondary cases in affected flocks meant that it is believed to be a poorly transmissible disease. This has led to the view that Atypical/Nor98 scrapie is a spontaneous disorder for which human and animal exposure risk remains low. In this study we demonstrate that in affected individuals, Atypical/Nor98 scrapie agent can disseminate in lymphoid tissues, nerves, and muscles, challenging the idea that it is a brain-restricted infectious agent. Evidence for the deficiencies in the current methods applied for monitoring Atypical/Nor98 scrapie is provided that would indicate an underestimation in the prevalence in the general population and in the affected flocks. These elements challenge the hypothesis on the biology of this recently identified TSE agent. According to the prion concept, abnormal PrP would be the causative agent of TSE Transmissible spongiform encephalopathies (TSE), or prion diseases, are fatal neurodegenerative disorders occurring in sheep (scrapie), cattle , or humans (Creutzfeldt-Jakob disease - CJD). The key event in TSE is the conversion of a normal cellular protein that contrasted with those normally observed in small ruminants TSE cases Prnp gene) In 1998 an Atypical/Nor98 Scrapie was identified in Norwegian sheep; the PrPCurrently Atypical/Nor98 Scrapie represents a significant part of the TSE cases identified in the EU small ruminant population, where its prevalence was estimated to range between 5 to 8 positive small ruminants per 10,000 tested per year Prnp) encoding for PrP protein Atypical/Nor98 scrapie cases have different biological features from those observed in other small ruminants TSE Sc detection generally correlates with the presence of infectivity Information about the tissue distribution of Atypical/Nor98 scrapie agent in the host species is limited Sc and infectivity in lymphoid tissues and central nervous system or resistance (homozygous and heterozygous ARR) to classical scrapie or BSE Atypical/Nor98 scrapie field cases (n\u200a=\u200a7) collected in three different countries were investigated for the presence of PrPs system . These sSc accumulation could be detected in different brain areas by WB and/or Immunohistochemistry (IHC). Conversely, no abnormal PrP deposits were evidenced in any of the investigated lymphoid organs Sc, the lymphoid tissue homogenates from five out of the seven cases were positive for TSE in tg338 mice. The attack rate in mice challenged with lymphoid tissues was lower and the clinical onset was delayed compared to mice inoculated using CNS homogenate were identical to the one observed with natural Atypical/Nor98 scrapie isolates and an AFRQ/ARQ (case 8) sheep which had been intra-cerebrally challenged with an AFRQ/AFRQ field Atypical/Nor98 scrapie isolate case 1, lane 5. detected . In the th cases . Incubatisolates .tg338 mice , which can be distinguished on the basis of their lesion profile in 338 mice . The ino samples . For bot pattern , vacuola pattern observedSc detection WB (TeSeE WB Kit \u2013 BIORAD), a PrPSc ELISA detection assay (TeSeE Sheep and Goat - BIORAD) and Atypical/Nor98 scrapie brain homogenates dilution series were prepared and processed for a OIE registered PrP BIORAD) and bioa338 mice .6.8 ID50 IC tg338 per gram and 106.6 ID50 IC tg338 per gram which corresponded to 102.2 (Langlade isolate) and 102 (PG127 isolate) ID50 IC tg338 which coin tg338 . SimilarIC tg338 . These rtg338 mice with the infectious dose were estimated respectively 10338/gram and 2. T338/gram .2,7 ID50 IC per gram of tissue); ie about 106 fold lower than the infectivity level measured (by endpoint titration) or estimated (on the basis of the incubation periods) in the same amount of brain prepared from clinically affected sheep affected with two different classical scrapie isolates did reach up to 1/10 (weight/weight) of the infectivity found in the CNS from terminally affected sheep. These values are higher than those expected from previous work. This could be explained by the fact that previously available data on prion quantities in peripheral tissues of small ruminants (in particular those related to striated muscle) relied on biochemical measurement of PrPThe classical scrapie cases that were investigated in this work cannot be assumed to be representative of all field diversity as only four animal cases of highly susceptible genotypes were used. However, the results indicate that exposure risk to such TSE agents through the unrestricted entry in the food chain of potentially infectious tissues would be significantly higher than previously thought.Sc presence in brainstem of a fraction of fallen or healthy culled small ruminants Sc detection tests that are used for initial field screening or confirmation of TSE cases is debated. Several authors reported failure to detect PrPSc in some CNS areas like the obex area In most countries, the identification of Atypical/Nor98 scrapie was a consequence of the implementation of an active surveillance for TSE consisting in random testing for PrPSc detection and bioassay indicated that CNS samples that would contain up to 107.4.\u2013107.7 ID50/g of Atypical/Nor98 scrapie (according to tg338 IC bioassay) could remain negative for PrPSc detection. In field, Atypical/Nor98 scrapie cases (Sc positive WB was observed in CNS samples in which infectious titre was estimated (on the basis of incubation period) to be higher than 105.8 ID50/g IC in tg338. Such discrepancies might reflect an individual variability of the PrPSc WB detection limits between atypical scrapie cases. It might alternatively be the consequence of a relative imprecision in estimating the titre of low infectious doses by the incubation period bioassay method.The results obtained in this study by comparing the analytical sensitivity of biochemical PrPie cases PrPSc poSc detection sensitivity limit was about 102 ID50 IC in tg338 (ie a tissue with a titre of 103.7 ID50/g IC in tg338). These differences strongly support the contention that diagnostic assays based on PrPSc detection have lower performance for identifying Atypical/Nor98 scrapie cases than classical scrapie cases. It is consequently highly probable that a significant number of Atypical/Nor98 cases remain undetected by field testing, leading to an underestimation of Atypical/Nor98 scrapie prevalence in the small ruminant population. It is however not possible on the sole basis of this study to evaluate the importance of such underestimation.In contrast to Atypical/Nor98 scrapie cases, using two different classical agents the WB PrPSc based approach would also impact on understanding of the biology of this TSE agent.The under detection of Atypical/Nor98 scrapie in the field due to the sensitivity of the current PrPSc based methodologies means that this conclusion should be considered with caution.While under natural conditions, classical scrapie is known to transmit between individuals, the analysis of data collected through the active TSE surveillance program seemed to indicate that Atypical/Nor98 scrapie could be poorly or not transmissible at all. This is based on the lack of statistical difference of the observed Atypical/Nor98 frequencies between the general population and the flocks where a positive case had been identified Sc detection in peripheral tissues of reported cases suggested that Atypical/Nor98 scrapie agent could be restricted to CNS. This is supportive of the hypothesis that Atypical/Nor98 scrapie could be a spontaneous disorder of PrP folding and metabolism occurring in aged animals without external cause Atypical/Nor98 cases are identified in older animals in comparison to classical scrapie Sc testing result could be observed in animals harbouring high infectious titre in their brain and that the infectious agent can be present in peripheral tissues of Atypical/Nor98 scrapie incubating sheep. TSE are considered to be transmitted following oral exposure; initial uptake is followed by a peripheral replication phase which is generally associated with a dissemination of the agent in the lymphoid system and the deposition of large amounts of PrPSc. This peripheral replication phase is later followed by the entry of the infectious agent into the CNS through the autonomic nervous system Sc based assays would explain the apparent old age of detected cases.However, this hypothesis is questioned by the evidence reported here that a negative PrPThe results presented here are insufficient to rule out the hypothesis of a spontaneous/non contagious disorder or to consider this alternative scenario as a plausible hypothesis. Indeed, the presence of Atypical scrapie/Nor98 infectivity in peripheral tissues could be alternatively due to the centripetal spreading of the agent from the CNS. However, our findings point out that further clarifications on Atypical/Nor98 scrapie agent biology are needed before accepting that this TSE is a spontaneous and non contagious disorder of small ruminants. Assessing Atypical/Nor98 scrapie transmissibility through oral route in natural host and presence in placenta and in colostrum/milk The presence of infectivity in peripheral tissues that enter the food chain clearly indicates that the risk of dietary exposure to Atypical/Nor98 scrapie cannot be disregarded. However, according to our observations, in comparison to the brain, the infectious titres in the peripheral tissues were five log10 lower in Atypical/Nor98 scrapie than in classical scrapie. Therefore, the reduction of the relative exposure risk following SRM removal is probably significantly higher in Atypical/Nor98 scrapie cases than in classical scrapie cases. However, considering the currently estimated prevalence of Atypical/Nor98 scrapie in healthy slaughtered EU population Finally, the capacity of Atypical/Nor98 scrapie agent to cross species barrier that naturally limits the transmission risk is insufficiently documented. Recently, the transmission of an Atypical/Nor98 scrapie isolate was reported into transgenic mice over-expressing the porcine PrP All animal experiments were performed in compliance with our institutional and national guidelines, in accordance with the European Community Council Directive 86/609/EEC. The experimental protocols were approved by the INRA Toulouse/ENVT and by the Norwegian ethics committees.The natural classical scrapie case (case 10) included in this experiment was a Romanov sheep born and bred in the Langlade flock where a natural scrapie epidemic has been occurring at a high incidence since 1993 Prnp gene as previously described Natural atypical scrapie cases were identified though active or passive surveillance programs in France, Norway and Portugal . The PorTwo sheep (one 12 months old AFRQ/ARQ (case 8) and one 14 months old AHQ/AHQ (case 9)) selected in a field flock were IC challenged with French AFRQ/AFRQ Atypical Scrapie (case 1) . The aniTSE-free Poll-Dorset sheep (VLA- Weybridge- UK) were used for intracerebral inoculation with Langlade isolate , or PG127 isolate . Animals were killed when displaying evident clinical signs at respectively 380 days and 160 days post inoculation. Oral challenge was performed in 6\u201310 months old TSE-free New Zealand cheviot sheep. Animals were dosed with 5 g equivalent of brain material (1% brain homogenate in glucose) derived from an experimentally VRQ/VRQ affected sheep (PG127 isolate). Animals were culled at clinical stage of the disease (200 days post inoculation).All tissues were collected using disposable equipment . The different field and experimental cases were sampled on different dates and/or places. Different instrument sets and containers were used for collecting, transporting and storing each sample. Finally, in all cases, peripheral tissues were collected before CNS to further reduce the risk of cross contamination. In natural Atypical/Nor98 cases, the nature of the tissues collected under TSE sterile conditions might have varied according to the country and date of collection. In all cases, CNS and at least one lymph node were available. In both Atypical/Nor98 and classical scrapie experimental cases, a large panel of tissues was collected under TSE sterile conditions.From the available samples, tissues homogenates were prepared in Norway (Norwegian cases) or in France . The list of processed samples is given in Sc IHC detection was first performed using 8G8 antibody raised against human recombinant PrP protein and specifically recognising the 95\u2013108 amino acid sequence (SQWNKP) of the PrP protein.This method was performed as previously described For each sample a negative serum control was included, in which the primary antibody was either omitted or replaced by purified mouse IgG2a serum.Sc extraction. The obtained pellet was denaturated in Laemmli's buffer (15 \u00b5l) before being loaded neat or diluted was used following the manufacturer's recommendations. For each sample, 250 \u00b5l of 10% brain homogenate were submitted to PrP diluted on a 12%sc was recovered as a pellet after addition of 500 \u00b5L of buffer B and centrifugation for 5 min at 20 000 g at room temperature. Supernatant was discarded and tubes dried. Finally, the pellet was denatured in buffer C (5 min at 100\u00b0C) and 1:6 diluted in R6 reagent before distribution into the wells A commercially available TSE detection test (TeSeE Sheep and Goat - BioRad) was used according to manufacturer's recommendations. In summary, five hundred \u00b5L of the 20% homogenate were incubated for 10 min at 37\u00b0C with 500 \u00b5L of buffer A containing proteinase K. PrPPET blots were performed using a method previously described tg338), which are considered to be highly efficient for the detection of sheep scrapie infectivity Bioassay experiments were carried out in ovine VRQ PrP transgenic mice in 5% glucose sterile solution. Each homogenate was then tested for bacteria presence (blood gelose overnight 37\u00b0C culture) and non sterile homogenates were submitted to a heat treatment (60\u00b0C \u2013 10 min). Heat treated samples are identified in Sc Western blot testing (TeSeE WB kit- BioRad) and the other part of the brain was formalin fixed for vacuolar brain lesion profiling Sc PET-Blotting.Portuguese and French cases' inoculations were carried out in UMR INRA ENVT 1225 facilities while Norwegian cases were inoculated at the NVI . Peripheral tissues and CNS homogenates were inoculated on different days in order to avoid any risk of cross contamination. In some cases, tissues autolysis resulted in the death of some animals inoculated which explain the low number of mice for some isolates. Mice were monitored daily until the occurrence clinical signs of TSE. Mice were culled when they started to show locomotor disorders and any impairment in their capacity to feed. CNS samples were individually collected. A part of the brain was frozen for PrPg338 mice, by inoculating intra-cerebrally (20 \u00b5l) successive 1/10 dilutions of CNS homogenate in groups of tg338 mice (6 or 12 mice). The material used for the titration was 10% brain homogenate except for the Langlade isolate (12.5% homogenate).Five different isolates were endpoint titrated in tTwo classical scrapie inocula used for sheep inoculation were titrated in UMR INRA ENVT 1225. The titration of the Langlade material was already published in a previous study et al. study in which they were respectively identified as Lindos and DS8 Two confirmed atypical scrapie isolates were titrated in INRA Jouy-en-Josas. These isolates correspond to two atypical cases originally described in the Le Dur Three additional atypical scrapie isolates were titrated in UMR INRA ENVT 1225.The infectious titre (Infectious Dose 50) of the brain homogenates was determined by the Spearman-K\u00e4rber's method tg338 and the number of ID50 inoculated to each mice (number of ID50 per 20 \u00b5L of the inoculated homogenate) were prepared by successive dilutions in negative brain homogenate.\u22123, 10\u22124, 10\u22125, 10\u22126, 10\u22127, 10\u22128. The dilutions series were tested for PrPSc using TeSeE Sheep and Goat ELISA test and the WB as previously described in the text.The prepared dilutions were: 1/2, 1/5, 1/10, 1/20, 1/40, 1/80, 1/100, 1/200, 1/400, 1/800, 10\u22125 to 10\u22127 dilutions from the same series were inoculated in groups of 6 tg338 in order to assess the infectious titre .The neat sample and 10tg338 mice were then tested by WB (TeSeE WB kit \u2013 BioRad).Dilutions series of CNS homogenates were prepared from the Langlade scrapie (case 10: posterior brainstem) and PG127 (case 12: posterior brainstem) homogenates that were endpoint titrated in 338 mice . For theFigure S1Sc detection limit in Atypical/Nor98 scrapie isolates.PrP Dilution series from two atypical Atypical/Nor98 scrapie isolates were prepared in negative sheep brain homogenate. The tissue homogenates (see methods) are the same than those used for endpoint titration in tg338 mice (A\u2013B) The samples were processed for PrPSc using an OIE registered Western-blotting technique . After extraction the re-suspended pellet was entirely loaded on gels. (A) case 1: cerebral cortex. (B) case 8: cerebellum. The same classical scrapie control than in Lane 1). (C) The same dilution series were tested using a commercially available rapid TSE test (TeSeE Sheep and Goat - BioRad) used for field TSE screening in small ruminants. Three different aliquots of each dilution were independently tested. The extractions were carried out. Results are presented as the mean +/\u2212 SD corrected optical density values. The cut off value (0.162 OD\u2013 dotted line) was established as the mean of four negative control optical densities +0.150 OD. (\u25cb: case 1. \u25b3: Case 8).338 mice . (A\u2013B) T(0.43 MB TIF)Click here for additional data file.Figure S2Sc Western Blot profile and vacuolar lesion profile in tPrPg338 mice inoculated with various tissues from an Atypical/Nor98 scrapie and classical scrapie affected sheep. (A) PrPSc Western Blot profile. Lane 1: posterior brainstem from a PG127 classical scrapie affected sheep (case 12). Lane 2: brain from a 700 days old negative control tg338 mouse. Lane 3: cerebral cortex from an AFRQ/AFRQ Atypical scrapie/Nor98 natural case (case 1). Lanes 4\u20137, brains from tg338 mice inoculated with various tissues homogenates prepared from an AFRQ/ARQ Atypical scrapie/Nor98 experimental case (case 8). Lane 4: cerebellum. Lane 5: prescapular lymph node. Lane 6: brachial nerve. Lane 7: external ocular motor muscle. Lane 8: brain from a tg338 mouse inoculated from a PG127 classical scrapie affected sheep (case 12). (B) PrPSc Western Blot profile. Lane 1: posterior brainstem from a PG127 classical scrapie isolate sheep (case 12). Lane 2: brain from a 700 days old negative control tg338 mouse. Lanes 3\u20139: brains from tg338 mice inoculated with various tissues homogenates prepared from two VRQ/VRQ PG127 classical scrapie cases (case 12 and 13). Lane 3: case 12, posterior brainstem. Lane 4: case 12, ileal lymph node. Lane 5: case 12, external ocular muscle. Lane 6: case 12, psoas muscle. Lane 7: case 13, posterior brainstem. Lane 8: case 13, tonsil. Lane 9: case 13, external ocular motor muscle. Lane 10: posterior brainstem from a PG127 classical scrapie sheep (case 12). (C\u2013D) Lesion profile (vacuolar changes) in tg338 mice inoculated with homogenates prepared from different tissues (C) an experimental Atypical/Nor98 AFRQ/ARQ scrapie case (case 8) or (D) an experimental VRQ/VRQ PG127 classical scrapie experimental case (case 12). (C) \u25cb cerebellum \u25c6: brachial nerve, \u25bd: external ocular motor muscle. (D) \u25cf: posterior brainstem, \u25bd: ileal lymph node, \u25cb: external ocular motor muscle.(0.53 MB TIF)Click here for additional data file."} +{"text": "Vitis vinifera L. such as Sultanina produce berries with seeds reduced in size. Stenospermocarpy has not yet been characterized at the molecular level.Stenospermocarpy is a mechanism through which certain genotypes of Vitis vinifera L. to improve QTL analysis for seedlessness and berry size in experimental progeny derived from a cross of two seedless genotypes. Major QTLs co-positioning for both traits on chromosome 18 defined a 92-kb confidence interval. Functional information from model species including Vitis suggested that VvAGL11, included in this confidence interval, might be the main positional candidate gene responsible for seed and berry development.Genetic and physical maps were integrated with the public genomic sequence of VvAGL11 at the sequence level in the experimental progeny identified several SNPs and INDELs in both regulatory and coding regions. In association analyses performed over three seasons, these SNPs and INDELs explained up to 78% and 44% of the phenotypic variation in seed and berry weight, respectively. Moreover, genetic experiments indicated that the regulatory region has a larger effect on the phenotype than the coding region. Transcriptional analysis lent additional support to the putative role of VvAGL11's regulatory region, as its expression is abolished in seedless genotypes at key stages of seed development. These results transform VvAGL11 into a functional candidate gene for further analyses based on genetic transformation.Characterization of For breeding purposes, intragenic markers were tested individually for marker assisted selection, and the best markers were those closest to the transcription start site.VvAGL11 is the major functional candidate gene for seedlessness, and we provide experimental evidence suggesting that the seedless phenotype might be caused by variations in its promoter region. Current knowledge of the function of its orthologous genes, its expression profile in Vitis varieties and the strong association between its sequence variation and the degree of seedlessness together indicate that the D-lineage MADS-box gene VvAGL11 corresponds to the Seed Development Inhibitor locus described earlier as a major locus for seedlessness. These results provide new hypotheses for further investigations of the molecular mechanisms involved in seed and berry development.We propose that Vitis vinifera L genomic resources, including both released genomic sequences C[AC][AT][GC][AC]A[CT][CG][CA]A[CG], with 7 (Vv) and 2 (At); and C[AT]CAT[CT]TC[TC][CA][AC], with 9 (Vv) and 3 (At). The first (and more abundant) motif corresponds to (GAGA)n putative regulatory elements, which are the binding site for BASIC PENTACYSTEINE1 (BPC1), a regulator of the homeotic Arabidopsis thaliana gene AGL11, which controls ovule identity where M is the phenotypic mean of the genotypes ; if d < 0, the a allele presents incomplete dominance over the b allele.The dominance effect Acquaah as follohttp://urgi.versailles.inra.fr/cmap) and sequences from the 6X genome assembly ; Lilium longiflorum, LMADS2, [GenBank:AAS01766]; A. thaliana, AG [GenBank:NP_567569], SHP1 [GenBank:NP_191437.1], SHP2 [GenBank:NP_850377.1] and AGL11 [GenBank:NP_192734.1]; P. hybrida, FBP7 [GenBank:CAA57311.1] and FBP11 [GenBank:CAA57445.1]; V. vinifera, VvMADS5 [GenBank:AAM21345.1], Sultanina Seedless and Seeded-derived alleles of VvAGL11 [GenBank:CAO1637]; Lilium longiflorum, LMADS2 [GenBank:AAS01766]; Gossypium hirsutum [GenBank:AAN15183]; Cucumis sativus [GenBank:AAC08529]; Lotus corniculatus [GenBank:AAX13306], Malus \u00d7 domestica [GenBank:CAA04324]; Prunus persica [GenBank:ABQ85556] and Prunus dulcis [GenBank:AAY30856].Click here for fileVvAGL11Predicted cis-regulatory elements that differ between seeded (pSEEDED) and seedless (pSEEDLESS) putative minimal regulatory region of . Both sequences were aligned on the genome reference sequence (pPN40024). SNPs and INDELs are signalled by coloured bases or sequence gaps. Yellow and blue segments represent 5'UTRs and TATA-box, putative cis-regulatory elements identified by PLACE database are indicated with brown segments with their respective accession number (last three digits). Red segments represent the polymorphic markers mapped in the RS \u00d7 S experimental progeny.Click here for filePredicted cis-regulatory elements identified by PLACE database that differ between the seeded and seedless putative minimun regulatory region (430 bp upstream the transcription start site) and the first intron (1.4 kb of the 5'UTR intron). Seeded and Seedless sequenced Sultanina-derived alleles were aligned and scanned for motifs by PLACE database.Click here for fileTranscript differences between seeded and seedless alleles derived from the RS \u00d7 S progeny. Nucleotidic and amino-acidic sequences from seedless (SEEDLESS cDNA VvAGL11) and seeded (SEEDED cDNA VvAGL11) alleles were aligned and compared against the predicted CDS from PN40024 . SNPs and non-silent mutations are signalled by coloured nucleotides or amino acids. Exons are represented by grey segments and size is in bp relative to the ATG.Click here for fileVvAGL11 exon 7 in a collection of Vitis vinifera genotypes maximizing sequence diversity and a few Vitis speciesNucleotide diversity of . Both already known seedless and seeded alleles from Ruby Seedless and Sultanina were included as well as Syrah (VvMADS5:SYH) and PN40024 (PNI). Exon 7 was obtained from a direct sequencing of PCR products using genomic DNA of the following genotypes as a template: cultivated Vitis vinifera such as Kishmish Chernyi (KIC), Asyl Kara (ASS), Orlovi Nokti Beli (ORL), Katta Kurgan (MAK), Araklinos (ARA), Arbois (ARB), Chardchi (CHB), Kapistoni Tetri (KAP), M\u00e9douar (MED), Mehdik (MEH), Oasis Bou Chemma 46 (OA7), Pletchistik (PLE), Tsitsa Kaprei (TIC), Tzolikoouri (TSO) and Lambrusque E (LAE), members of the Vitis genus such as Vitis berlandieri (VBE), Vitis aestivalis (VAE), Vitis coignetiae (VCO), Vitis labrusca (VLI) and Vitis rupestris (VRU), and one wild Vitis vinifera such as Lambrusque Sejnene 1 (LAS). Polymorphisms are signaled by colored nucleotides or amino acids. An asterisk signals seedless genotypes.Click here for fileVvAGL11 intragenic marker mapping and QTL analysis for seedlessness and berry size detected over three different seasons on chromosome 18. A: Consensus genetic map of chromosome 18 based on the RS \u00d7 S progeny. Green, pink and red markers correspond to SSRs developed in this study from Cabernet Sauvignon BAC End Sequence, from contig assemblies of the grapevine genome sequencing project, and from VvAGL11 allele sequencing, respectively. B and C: Projected seedlessness and berry size QTLs represented by coloured vertical bars and LOD (logarithm of the odds) profiles to the right of chromosome 18. Red, blue and green lines correspond to 2007, 2009 and 2010 seasons, respectively. Bar lengths are representative of their confidence interval once projected on the consensus map. Seedlessness was analyzed as seed fresh weight (SFW) and berry size as berry weight (BW). 1-LOD and 2-LOD support intervals were used for the prediction of the confidence intervals. Vertical dashed line in the LOD profile represents the LOD threshold for significant QTLs according to the permutation tests. Genetic distances are expressed in centimorgans (cM).Click here for fileAssociation analysis performed in a population derived from crosses of several seedless varieties. A population (N = 146 seedlings) originating from 14 progeny derived from crosses of 11 seedless varieties was genotyped with intragenic marker p3_VvAGL11, and mean seed fresh weight per berry was recorded. Allele sizes were determined by capillary electrophoresis. Association analysis was performed by one-way ANOVA, and different letters represent significant differences at P < 0.05 by Fisher's least significant difference procedure.Click here for fileGenetic and physical distance between markers comprised within the confidence interval. Microsatellite repeat and segregation type, relative position in linkage map, distance between loci, position in the reference genome assembly, physical distance between loci and recombination frequency between adjacent markers is indicated. Underlined markers belong to the confidence interval from the major QTL for seedlessness reported in this work.Click here for filePrimer pairs designed to sequence the regulatory region of VvAGL11. Oligos were designed by Primer3Plus web interface using PN40024 sequence as template.Click here for file"} +{"text": "Whole body immersion in water (WI) constitutes a significant role in the area of CAM as well as in rehabilitation facilities. WI has strong effects on the autonomic nervous system. In this study, we investigate the effects of different water temperatures on heart rate variability (HRV), peripheral and core body temperature .Twenty-one healthy subjects underwent WI with water temperatures of 33\u00b0C (WI33), 36\u00b0C (WI36), 39\u00b0C (WI39). The procedure was: supine rest (30 minutes), WI (20 minutes) and supine rest (30 minutes). An ECG, the nasal/oral airflow, core body and temperature of extremities were recorded. The results of the last 5 minutes at the end of each interval are presented.During WI33 and WI36 CBT decreased compared to rest before WI, whereas WI39 led to an increase of CBT. Peripheral temperature was determined by the water temperature. The average RR-interval increased during WI33 (970 ms) compared to rest before WI (910 ms), whereas it decreased during WI36 (850 ms) and WI39 (636 ms). The standard deviation of the RR-intervals (SDNN) was reduced during WI39 (26 ms), whereas it was augmented during WI33 (78 ms) compared to rest before WI (63 ms). The square root of the mean squared difference of successive RR-intervals (RMSSD) decreased during WI39 (14 ms) and WI36 (43 ms), whereas it increased during WI33 (72 ms) compared to rest before WI (52 ms).Each water temperature showed specific effects on CBT, PT and HRV during WI. WI39 lead to an increase of CBT and PT and a decrease of HRV whereas WI33 showed opposite effects. Hence, WI39 induces moderate cardiovascular stress and moderate hyperthermia whereas WI33 induces mild hypothermia and cardiovascular relaxation. The water temperature is crucial for therapeutic purposes."} +{"text": "Atherosclerotic cardiovascular diseases are the leading causes of morbidity and mortality worldwide. We have previously shown that arachidonate 15-lipoxygenase B (ALOX15B) is highly expressed in atherosclerotic carotid plaques, and elucidation of mechanisms downstream of activated lipoxygenases may be relevant to our understanding of the genesis of atherosclerotic diseases. We examined 120 carotid plaques from patients with symptomatic carotid artery stenosis and showed that the extent of ALOX15B staining was significantly increased in carotid plaques with thrombosis. Impedance aggregometry analyses showed that the ALOX15B enzyme products 15-HETE and 15-HPETE increased platelet aggregation. By using a calibrated automatic thrombin assay, we showed that the ALOX15B products also increased both peak levels of thrombin and the total endogenous thrombin potential. Moreover, platelet aggregation was increased by addition of cell lysates from ischemic human macrophages, whereas platelet aggregation was reduced after knockdown of ALOX15B in human macrophages. Our data show that ALOX15B expression in human carotid plaques is associated with thrombus formation and that enzyme products of ALOX15B increase platelet aggregation and thrombin generation. We therefore propose that activated ALOX15B in macrophages may play a role in the induction of atherothrombotic events by increasing platelet aggregation and thrombin generation. Atherosclerotic lesions are common in the carotid artery, but only a minority of the plaques ever causes cerebrovascular ischemic events. Symptomatic carotid atherosclerotic plaques and rupture-prone vulnerable plaques have histopathological features such as inflammatory monocytes/macrophages, cell infiltration and a thin fibrous cap leading to hemorrhage. Chronic inflammation is evident at every stage from initiation to progression and eventually to plaque rupture and thrombosis 2 increase platelet aggregation Inflammation in the vascular wall and thrombosis are linked processes, and cellular activation with production of proinflammatory molecules may also be prothrombotic Here, we studied carotid plaques from patients who suffered an ischemic event caused by carotid atherosclerosis and investigated if ALOX15B expression in these carotid plaques is associated with thrombus formation. We also investigated how the ALOX15B enzyme products 15-HETE and 15-HPETE affect platelet aggregation.http://www.wlab.gu.se/bergstrom/guvasc/). Patient characteristics are shown in Carotid endarterectomies from 120 patients with high-grade symptomatic carotid artery stenosis , mouse monoclonal anti-CD68 , CD42 mouse monoclonal antibody . Detection was performed with Mach3 kit and Vulcan Fast Red . Stained sections were digitalized using a Zeiss Mirax Scanner . Digital images were analyzed using the BioPix software . The extent of immunohistochemical staining is expressed as a percentage of the stained area of the total section area.Thrombus formation was classified using the following criteria: rupture of the fibrous cap with clear communication between the necrotic core and the lumen and adjacent surface thrombus .Histopathological classification of plaques was made on deparaffinized serial sections stained with Mayers hematoxylin end eosin according to the American Heart Association (AHA) classification The study protocol was approved by the Ethical Committee of the University of Gothenburg (Dnr.404\u201309) and all subjects gave written informed consent.Buffy coats were obtained from 8 healthy adult volunteer blood donors at Kung\u00e4lv Hospital, Sweden, and samples were de-identified before handling. Human mononuclear cells were isolated by centrifugation in a discontinuous gradient of Ficoll-Paque . Cells were seeded in Macrophage-SFM medium (Gibco) containing granulocyte macrophage colony stimulating factor (GM-CSF). After 3 days, the medium was changed to RPMI 1640 medium without GM-CSF and cells were cultured for 7 days before further experiments. To test the effect of ischemia, cells from 4 blood donors were incubated in 21% or 1% oxygen for 24 hours. To test the effect of ALOX15B knockdown, cells from a further 4 blood donors were incubated in 1% oxygen and transfected with 20 nmol/L ALOX15B siRNA or nonsilencing control siRNA in HiPerFect transfection reagent (Qiagen) according to the manufacturer's recommendations. Cells were washed after 24 hours, siRNA was added again and cells were incubated at 1% oxygen for 24 hours before extraction of RNA.Total RNA was isolated with the RNeasy kit (Qiagen). Expression of human ALOX15B mRNA was determined and normalized to \u03b2-actin mRNA expression using quantitative real-time PCR. The reverse transcription reaction was set up using a cDNA reverse transcription kit (#4368814) and performed with a Gene Amp PCR system 9700 (Applied Biosystems). Real-time PCR amplification was set up using TaqMan gene expression assays for human ALOX15B (Hs00153988_m1), human actin B (Hs99999903_m1) in combination with Universal PCR master mix (#4324018) and performed for 50 cycles on an ABI PRISM 7700 sequence detection system (Applied Biosystems).Cell lysates for platelet aggregation and tissue factor measurements were prepared by adding lysis buffer containing 0.15 mmol/l NaCl, 10 mmol/L Tris-HCl, 2 mmol/L EDTA and 1% TritonX-100 with protease inhibitors . Cell lysates for measurement of thrombin generation and 15-HETE analysis were prepared by adding cold PBS supplemented with protease inhibitors (Complete Mini) to the macrophages, cell were lysed by freezing at -20\u00b0C and thawing four times before analysis.15-HETE was analyzed in cell lysates by 15-HETE ELISA kit . Tissue factor was analyzed in cell lysates by Human Coagulation Factor III/Tissue Factor Quantikine ELISA kit .\u00ae Platelet Function Analyzer as described Whole blood platelet aggregation was measured using the MultiplateThe calibrated automated thrombin\u00ae (CAT) assay was used to study the effect of 15-HETE and 15-HPETE on thrombin peak level and the total endogenous thrombin potential as previously described Data are plotted as mean and SEM unless stated otherwise. All analyses were performed using GraphPad Prism version 5.01 for Windows ; a 95% confidence interval was used and P values <0.05 were considered significant. Correlations between groups were determined using non-parametric two tailed correlation (Spearman two tailed correlation). Differences between groups were determined using non-parametric two tailed t-test (Mann-Whitney two tailed t-test) or ANOVA.Patient and plaque characteristics are shown in s\u200a=\u200a0.45, P<0.0001), as expected, and was higher in carotid plaques with thrombosis compared with no thrombosis of 15-HETE and 15-HPETE. Both 15-HETE and 15-HPETE increased platelet aggregation in vitro . BecauseTo investigate if ALOX15B enzyme products secreted from ischemic macrophages increase platelet aggregation, we incubated macrophages in a low oxygen concentration to mimic ischemia and activate ALOX15B enzyme activity. We first confirmed our earlier findings that ischemia increases ALOX15B mRNA expression and 15-HETE levels . We alsoTo specifically investigate the importance of ALOX15B in platelet activation and thrombin generation, we transfected human primary macrophages with control siRNA and ALOX15B siRNA. Knockdown of ALOX15B resulted in a reduction of mRNA levels to 55% of non-silenced levels and signThe present study showed that ALOX15B expression in human carotid plaques was associated with thrombus formation in the plaque, and high expression of ALOX15B was found in patients diagnosed with stroke. The ALOX15B enzyme products 15-HETE and 15-HPETE increased platelet aggregation and thrombin generation in vitro. Further analysis showed that knockdown of ALOX15B in primary human macrophages decreased 15-HETE production, platelet aggregation and thrombin generation.It is clear that atherosclerosis is a chronic inflammatory condition that can lead to an acute clinical event by plaque rupture and thrombosis Previous studies have reported associations between high ALOX15B expression in carotid lesions and a history of cerebrovascular symptoms 2 is regarded as one important agonist in this process Platelet aggregation measurements are used to predict the occurrence of thrombotic events following percutaneous coronary interventions Our previous results showed that overexpression of ALOX15B in human macrophages resulted in increased secretion of 15-HETE Tissue factor triggers blood coagulation and is expressed in atherosclerotic plaques; it may therefore contribute to thrombus formation after plaque rupture In summary, this study shows that ALOX15B staining associates with thrombosis in human carotid plaques, and that platelet aggregation and thrombin formation are increased by ALOX15B enzyme products. We therefore propose that activated ALOX15B in carotid plaques plays a role in the induction of atherothrombotic events by increasing thrombin generation and platelet aggregation.Figure S1Immunohistochemical detection of thrombus formation in carotid endarterectomy specimen. (A) Complicated atherosclerotic plaque with a thin fibrous cap and a plaque rupture (arrows) exposing the underlying necrotic core (NC). Thrombus (T) formation is seen in the region of the plaque rupture. Scale bar 1000 \u00b5m. (B) Thrombus formation was classified using Haematoxylin & Eosin staining using the following criteria in high-magnification: Rupture of the fibrous cap (arrows) with clear communication between the necrotic core (NC) and the lumen (L) and adjacent surface thrombus (T). Scale bar 200 \u00b5m. (C) High-magnification on CD42 staining visualizing the thrombus (T) in plaque rupture region (arrows). Scale bar 200 \u00b5m.(TIF)Click here for additional data file.Figure S2Ischemia or ALOX15B knockdown had no effect on tissue factor levels in human macrophages. (A) Primary human monocyte-derived macrophages from 4 blood donors were cultured for 24 hours in in 21% oxygen (Control) and in 1% oxygen (Ischemia). Tissue Factor was analyzed in cell lysates by using Human Coagulation Factor III/Tissue Factor ELISA kit . Data shown are representative of 4 independent experiments analyzed in duplicates; mean \u00b1 SEM. (B) Human primary macrophages from 4 blood donors transfected with non-silencing control siRNA or ALOX15B siRNA and incubated for 24 hours in ischemia. Tissue Factor was analyzed in cell lysates by using Human Coagulation Factor III/Tissue Factor ELISA kit (R&D Systems Europe Ltd.). Data shown are representative of 4 independent experiments analyzed in duplicates; mean \u00b1 SEM.(TIF)Click here for additional data file."} +{"text": "The aim of this study was to evaluate the clinical significance of unexpected focal FDG uptake within the breast in patients undergoing PET-CT for assessment of other malignancies.Consecutive adult patients undergoing FDG PET-CT for assessment of a nonbreast primary cancer between February 2009 and October 2012 were retrospectively reviewed. The incidence of focal FDG uptake within the breast was determined. PET parameters including maximum standardised uptake value (SUVmax), metabolic tumour volume, and total lesion glycolysis were recorded for each patient. The presence and patterns of morphologic changes on CT were assessed. Aetiology and clinical significance were confirmed histologically or by imaging and clinical follow-up.A total 23 of 8,972 patients (0.25%) had unexpected focal FDG uptake in the breast. Twenty patients (86%) underwent biopsy. Malignancy was confirmed in 17 patients, five patients (22%) had disseminated disease from their primary cancer and 12 patients (52%) had an unsuspected synchronous breast cancer. Eight patients (67%) with newly diagnosed breast cancer were outside the screening programme (\u226447 \u226573 years). Only five patients (42%) had a breast lesion with SUVmax >4. No PET parameters reliably distinguished benign from malignant pathology.Incidental focal FDG uptake in the breast is rare but requires further evaluation as approximately 50% of cases may reflect unsuspected breast malignancy."} +{"text": "Perilipin1, a lipid droplet associated protein has an important role in the regulation of lipolysis and lipid storage in adipocytes. Perilipin1 is also expressed in foam cells of atheroma plaques and could therefore play a role in the accumulation of lipids in arterial wall and in the development of atherosclerosis. The aim of the study was to investigate this possible role of perilipin1 in atherogenesis.Plin1-/-) were crossed with Ldlr-/- mice. Ldlr-/- and Plin1-/- Ldlr-/- mice received an atherogenic diet during 10 or 20 weeks. Blood pressure and plasma lipids concentrations were measured. Aortas were collected at the end of the atherogenic diet periods for quantification of atheroma lesions (en face method), histological and immunohistological studiesMice deficient in perilipin1 (Ldlr-/- and Plin1-/- Ldlr-/- mice had comparable blood pressure and plasma lipids levels. Plin1-/- Ldlr-/- mice had a lower body weight and decreased adiposity. The atherosclerotic lesion area in Plin1-/-Ldlr-/- mice was moderately increased after 10 weeks of atherogenic diet (ns) and significantly higher after 20 weeks (p < 0.01). Histology of atheroma plaques was comparable with no sign of increased inflammation in Plin1-/- Ldlr-/- mice.Perilipin1 ablation in mice results in increased atherosclerosis independently of modifications of risk factors such as raised blood pressure or plasma lipids levels. These data strongly support an atheroprotective role for perilipin1. A hallmark of atherosclerosis is the accumulation of free (FC) and esterified (EC) cholesterol in macrophages and smooth muscular cells transforming them in foam cells . Such acLdlr -/- mouse.Storage of lipids droplets in cells accumulating triacylglycerols or EC (steroidogenic cells) is also dependent in part of specific proteins surrounding these droplets and belonging to the PERILIPIN family (previously named PAT family) , particuPlin1-/- mice were a gift from I Tabas . These mice are from the strain generated and described by Martinez-Botas et al [Ldlr-/- mice (C57BL/6 background) were from the Jackson Laboratory . Plin1-/- and Ldlr-/- mice were crossed to obtain Plin+/-Ldlr +/- mice, which were intercrossed to obtain Ldlr-/- and Plin1-/- Ldlr-/- mice. All mice were housed in an animal facility with controlled temperature (21-23\u00b0C) and lighting and had free access to food and water. Male mice were used for the experiments. These mice received, starting at 8 weeks of age, an atherogenic diet and were investigated after 10 or 20 weeks of atherogenic diet. This study was approved by the Ethical Committee of the University Cl Bernard of Lyon.as et al and are Ldlr-/-and 16 Plin1-/- Ldlr-/- mice) by the tail-cuff method (Visitech 2000 series II) after acclimation to restrain and tail-cuff inflation. For quantification of atheroma mice were anaesthetized with pentobarbital. Blood was collected from inferior vena cava and plasma separated by centrifugation for enzymatic determination of total cholesterol and of triacylglycerols concentrations. Aortas were dissected from aortic root to iliac bifurcation, carefully cleaned form periarterial adipose tissue, pinned on silicon dishes and stained for lipid deposits with Red Sudan IV (en face method). Red Sudan IV positive areas were quantified using Image J software and expressed as the percentage of total aorta area. For histological studies some aortas of 20 weeks atherogenic diet fed mice were rinced with PBS and fixed in phosphate buffered formalin, carefully dissected, dehydrated and embedded in paraffin before sectioning. Haematoxylin-eosin and Verhoeff (elastic fibres) stainings were performed as well as immunocytology for macrophages (anti-Mac3 antibodies), lymphocytes (anti-CD3 antibodies) and smooth muscular cells (anti \u03b1-SMA antibodies) [Ldlr-/-and 5 Plin1-/- Ldlr-/-) were removed, flushed with cold isotonic saline, carefully cleaned of perivascular adipose tissue and flash frozen in liquid nitrogen before storage at -80\u00b0C until analysis. Total RNAs were purified using TRIZOLR protocol with the addition of a DNase treatment. Concentrations, intergrity and purity were verified. For measurements of individual mRNA levels , total RNA was reverse transcripted using Superscript II (Invitrogen) and random hexamers. Real time PCR was performed in a MyIQ thermal cycler using iQ SYBR green Supermix (Biorad). Samples were run along with dilutions of known amounts of target sequence for quantification of initial cDNA copies. Results were calculated as the target over 18S RNA concentration ratio (ng/\u03bcg). Primer sequences are shown in table Blood pressure and heart rate were measured in 20 weeks old mice (8 ibodies) . For meaResults are shown as individual values or as means \u00b1 sem. Comparisons between groups were performed by Student t test for non paired values or by Mann-Whitney test using GraphPad Prism (version 5.03). P < 0.05 was considered as indicating a significant difference.Plin1-/- Ldlr-/- and Ldlr-/- mice but did not differ between the two groups of mice (table Ldlr-/- mice) and heart rate (547 \u00b1 25 vs 515 \u00b1 23 b/min) were also comparable. Plin1-/- Ldlr-/- had a slightly lower body weight and as expected [Plin1-/- Ldlr-/- mice had after 10 weeks of atherogenic diet a trend for increase in atherosclerosis at 20 weeks. Figure Plin1-/- Ldlr-/- and Ldlr-/- mice. Histological examination of plaques performed on aortas from mice fed since 20 weeks the atherogenic diet showed no differences in structure between Plin1-/- Ldlr-/- and Ldlr-/- mice and only a non significant trend for higher values of TNF\u03b1 mRNA (1.72 \u00b1 0.37 10-4 vs 1.13 \u00b1 0.26 10-5 ng/\u03bcg 18S RNA p = 0.10) in aortas from Plin1-/- Ldlr-/- mice. The expression of SR-A, implicated in the uptake of modified lipoproteins, and of ABCA1 and ABCG1, controlling the efflux of cholesterol, were increased in aortas of Plin1-/- Ldlr-/- mice .Plasma cholesterol levels were high in both ce table . Plasma expected an evideLdlr-/- mice with perilipin1 ablation. The overall histological appearance of the plaques was unchanged and we observed no increase in macrophages or lymphocytes infiltration, nor any significant increase in the mRNA concentrations of pro-inflammatory cytokines and chemokines. Therefore inflammation does not appear increased and the enhanced atheroma seems to result mainly form an increase in lipid deposits. Such an increase is demonstrated by the examination of aortas by the en face method since Red Sudan IV stains lipid deposits. An increased expression of SR-A could contribute to this increase in lipid deposits. The increase in ABCA1 and ABCG1 expression would on the contrary favour cholesterol efflux and could be an adaptative response to the increased accumulation of intra-cellular cholesterol. Perilipin1 can be found in macrophages and vascular smooth muscular cells and is present within atheroma plaques in foam cells originating from both cell types [The present results support an atheroprotective role of perilipin1 since the extent of atheroma lesions was increased in ll types -22. We ull types . Perilipll types . We cannIn conclusion we found a protective role of perilipin1 against atherosclerosis. Evaluation of the respective roles of perilipin1 in macrophages and in smooth muscular cells and elucidation of the precise mechanisms implicated will require further studies.The authors declare that they have no competing interests.DL had in charge the mice strains and performed with JYL the quantification of atheroma and histological studies. FF and PDC had in charge mice strains and collected data, particularly plasma lipids and blood pressure. SN had in charge mRNA determinations. MB designed the study and wrote the manuscript. All authors participated in the analysis of data and the preparation of the manuscript."} +{"text": "A study of symptomatic breast units geographically spread over Ireland collected image quality, compression and radiation dose data from 18 mammography units; so how do these optimisation parameters compare nationally and internationally? The mean glandular dose (MGD) diagnostic reference level was proposed for the all-digital breast screening service [The quantitative and qualitative data were analysed using SPSS. Recommendations of MGD diagnostic reference levels were made at various levels for film-screen mammography (FSM) and full-field digital mammography (FFDM) units to match those levels published in worldwide.MGDs received by symptomatic breast patients within Ireland are higher than those received in the all-digital Irish Breast Screening service, although the differences for FFDM are not substantial; 55 to 65 mm breast: 1.75 mGy (screening) versus 2.4 mGy (symptomatic) at the 95th percentile. The four-view routine mammography MGDs obtained in symptomatic breast units in Ireland are, however, substantially different from other screening units with mixed FSM/FFDM modalities: 4.5 mGy (UK); 4.98 mGy (USA) versus 5.96 mGy and 9.63 mGy . Various reasons are proposed for the differences.MGD diagnostic reference levels achieved in the screening service may be lower due to the exacting requirements for radiographer training, nonsurgical alteration of patient breasts and equipment quality assurance levels. Greater training of radiographers performing mammography in the symptomatic breast services is required to standardise mammographic projections with regard to MGDs delivered."} +{"text": "There is large body of evidence on the correlation between chronic inflammatory diseases like rheumatoid arthritis (RA) and increased risk of cardiovascular diseases (CVD). Chronic inflammation is an independent risk factor for the development of early atherosclerosis. The possible role of anti TNF- \u03b1 biologic drugs in reducing this risk arouses great interest among the scientific community. Many studies indicate the possibility that anti-TNF-\u03b1 agents may reduce CVD risk and mortality in adult RA patients. There are only few data about the pediatric population.To determine the presence of early biomarkers of endothelial dysfunction in juvenile idiopathic arthritis (JIA) and to evaluate their improvement during anti TNF-\u03b1 treatment with etanercept.2\u03b1, total nitric oxide, TXB2 and PGE2).We enrolled 30 children affected by JIA, all eligible for anti-TNF-\u03b1 treatment. All patients were examined at baseline and after 6 months and 12 months of treatment with etanercept. Disease activity score for each patient was determined using JADAS (Juvenile Arthritis Disease Activity Score). Laboratory parameters included acute phase reactants (CRP and ESR), complete lipid profile, pro-inflammatory cytokines and biomarkers of oxidative stress and vascular inflammation and 8-iso-PGF2\u03b1 (a surrogate marker of oxidized LDL) showed significant improvement during treatment.During the study all biomarkers of endothelial dysfunction progressively improved after 1 year of etanercept treatment in JIA children. At the same time all inflammatory parameters and JADAS scores significantly improved when compared to baseline. Pro-inflammatory cytokines showed significant reduction both after 6 and 12 months of treatment; at 6 months a relative increase in TNF-\u03b1 and IL-6 was determined. Lipid profile, biomarkers of endothelial activation (total nitric oxide, PGEWe demonstrated beneficial effect of 1-year etanercept treatment on clinical disease activity, inflammatory indexes and oxidative biomarkers in JIA children. The role of inflammation on pre-atherosclerotic determinants justify the need of precocious interventions in JIA, in order to optimize the clinical outcome and to realize primary prevention of cardiovascular events in adulthood."} +{"text": "The developing zebrafish embryo has been the subject of many studies of regional patterning, stereotypical cell movements and changes in cell shape. To better study the morphological features of cells during gastrulation, we generated mosaic embryos expressing membrane attached Dendra2 to highlight cellular boundaries. We find that intercellular bridges join a significant fraction of epiblast cells in the zebrafish embryo, reaching several cell diameters in length and spanning across different regions of the developing embryos. These intercellular bridges are distinct from the cellular protrusions previously reported as extending from hypoblast cells (1\u20132 cellular diameters in length) or epiblast cells (which were shorter). Most of the intercellular bridges were formed at pre-gastrula stages by the daughters of a dividing cell maintaining a membrane tether as they move apart after mitosis. These intercellular bridges persist during gastrulation and can mediate the transfer of proteins between distant cells. These findings reveal a surprising feature of the cellular landscape in zebrafish embryos and open new possibilities for cell-cell communication during gastrulation, with implications for modeling, cellular mechanics, and morphogenetic signaling. The enveloping layer (EVL) is the outermost part of the blastoderm, surrounding the epiblast, the hypoblast and the yolk syncytial layer (YSL), which lay on top of the yolk mass [1]. DesTo study cell shape in the zebrafish epiblast, we generated high cellular contrast by targeting photoconvertible protein Dendra2 Three-dimensional imaging revealed that the cells linked by intercellular bridges resided within the epiblast layer; bridges were not observed linking cells in either the EVL or the hypoblast. A quantitative analysis of 51 intercellular bridges from 38 independent embryos at late gastrula stages showed that the protrusions averaged 215 \u00b5m in length see . The imaDrosophila melanogasterTime lapse studies of embryos show that the intercellular bridges form when a mitotic pair of daughter cells maintain a membrane tether, which is the lengthened as the cells move further apart and as neighboring cells subsequently intercalate between them . To docuThe intercellular bridges persisted throughout gastrulation , even asQuantitative analysis of the intercellular bridges during gastrulation showed that they lack any obvious orientation or preferential position. We superimposed bridges of 30 cell pairs from 20 different embryos at the mid-gastrula stage , using t2, almost identical to the surface area of the longest intercellular bridge of 1 \u00b5m diameter (1100 \u00b5m2). Thus, the surface area of these membrane bridges increases the amount of cell surface of the linked cells, and the total present in the embryo.Further characterization of intercellular bridges shows that there are minor variations in diameter (up to 1 \u00b5m) and significant variations in length, reaching lengths up to 350 \u00b5m and peripheral (mDendra2) membrane proteins along the intercellular bridges is fast enough to mediate cell-cell communication during gastrulation and neurulation. The formation, persistence, and material transfer along the intercellular bridges provide an unexpected pathway for the transfer of materials between cells in the zebrafish embryo. Their presence has important implications for not only membrane homeostasis, but also on the mechanical and signaling processes in embryonic patterning and development.Raising, maintaining, injection and spawning of wild-type AB zebrafish in house colony were performed as previously described cis-sequences necessary for transpositionmCD8) gene To generate embryos with high expression levels distributed in a mosaic subset of cells we used Tol2-mediated transient expression Injected Embryos were raised at 28\u00b0C in 30% Danieau solution Figure S1Intercellular Bridge Properties. (A) Animal pole view of a mosaic zebrafish embryo expressing Dendra2 at the onset of gastrulation. Note that intercellular bridges cannot be visualised even under intense 488\u00a0nm illumination of the cytoplasmic fluorescent protein. Intercellular bridges visualised at tailbud stage (B) and 3 somites stage (C). (D\u2013F) Intercellular bridge connecting two cells (white arrowheads) labelled with membrane targeted mCherry (D), EGFP-\u03b2-Actin (E), and overlay (F). Note that EGFP-\u03b2-Actin is partially present inside the intercellular bridge. Scale bar: 20 \u00b5m.(TIF)Click here for additional data file.Movie S1Intercellular Bridge Formation (related to ). Time-lapse imaging capturing the formation of an intercellular bridge after cell division at late blastula stage. Nuclei are labelled with H2B-mCherry (red) and cellular boundaries are labelled with mDendra2 (green). Scale bar 25: \u00b5m.(AVI)Click here for additional data file.Movie S2Intercellular Bridge Persistence During Gastrulation (related to). Time-lapse imaging showing the stability of an intercellular bridge (red arrowheads) labelled with mDendra2 (green) from mid-gastrula to the end of gastrulation. Scale of the grid: 25 \u00b5m.(AVI)Click here for additional data file.Movie S3Intercellular Bridge After Gastrulation (related to). Time-lapse imaging showing the presence of intercellular bridge labelled with mDendra2 (green) from 90% epiboly to 10 somites stage. Scale bar: 50 \u00b5m.(AVI)Click here for additional data file.Movie S4Intercellular Bridges and Cell Division During Gastrulation (related to). Time-lapse imaging showing a cell equipped with an intercellular bridge labelled with photoconverted mDendra2 (red) that undergoes oriented cell division. Scale bar: 25 \u00b5m.(AVI)Click here for additional data file.Movie S5Intercellular Bridge Fragmentation (related to). Time-lapse imaging showing the breakdown of an intercellular bridge labelled with mDendra2 by a migratory cell (white arrowhead). Scale bar 25 \u00b5m.(AVI)Click here for additional data file.Movie S6Dynamic Behaviour Along Intercellular Bridges (related to). Representative image sequences of the displacement of photoconverted mDendra2 (red) along the plasma membrane of an intercellular bridge. Scale bar: 25 \u00b5m.(AVI)Click here for additional data file."} +{"text": "Atazanavir (ATV) is administered at a dose of 300 mg boosted with 100 mg of ritonavir once daily (boosted). However, in the clinical setting use of unboosted ATV can be warranted. Although plasma concentrations are used clinically as a marker of drug exposure, ATV predominantly acts within HIV infected cells and therefore intracellular concentrations may better correlate with therapeutic efficacy. To date, the factors that define ATV flux into peripheral blood mononuclear cells (PBMC) are not fully characterized and several efflux and influx transporters may influence intracellular pharmacokinetics.To evaluate gene expression of efflux and influx (SLCO3A1) transporters and relate to the cellular accumulation of unboosted ATV.Patients administered with unboosted ATV were recruited in Torino. Written informed consent was obtained. Main inclusion criteria were: no concomitant interacting drugs (except for TDF), no hepatic or renal impairment and self-reported adherence > 95%. Blood samples were collected 22-26 h after dosing and PBMC and plasma separated. Plasma samples were analysed by a validated HPLC-PDA method and PBMC extracts (intracellular) analysed using a validated LC-MS method. The cell count and mean cellular volume (MCV) was used for determining intracellular concentrations. Gene expression was evaluated by relative quantification using real time PCR.13 Caucasian patients met the inclusion criteria and were included in the study. Median plasma ATV Ctrough was 134 ng/ml , intracellular concentrations were 322 ng/ml , and the median accumulation ratio (intracellular/plasma concentration) was 1.84 . Intracellular concentrations were not correlated with plasma concentrations (rho= -0.22 p=0.41). SLCO3A1 expression was significantly correlated with cellular accumulation ratio. . In multivariate linear regression analyses, SLCO3A1 expression was the only independent predictor of ATV cellular accumulation .Intracellular ATV concentrations were higher than plasma concentrations, indicating an accumulation of ATV in PBMC and potentially a role for influx transporters. The correlation between SLCO3A1 expression and ATV accumulation supports this hypothesis and suggests that the SLCO3A1 uptake transporter may be a determinant of intracellular ATV concentrations."} +{"text": "Left bundle branch block (LBBB) is a condition associated with increased mortality and poor prognosis, depending on the underlying etiology. Identification of the etiology and appropriate risk stratification of patients with LBBB is a challenge, despite the availability of various testing modalities. Nuclear exercise stress testing can show artificial perfusion defects in the interventricular septum. The artifacts are reduced but not completely excluded by use of vasodilator stress agents. In this study, we retrospectively reviewed the stress cardiac magnetic resonance (CMR) findings in patients with LBBB.We retrospectively reviewed 49 consecutive patients with complete LBBB who underwent stress CMR. Of these 49 patients, 23 also underwent concurrent coronary angiography, 20 underwent nuclear stress test and 8 underwent all the three tests. The reference standard for flow limiting coronary artery disease (CAD) was coronary artery stenosis greater than 50% on coronary angiogram.Stress CMR correctly identified all patients (n=13) with more than 50% coronary artery stenosis and all patients (n=7) without significant coronary artery stenosis.Overall, the sensitivity, specificity, positive predictive value and negative predictive value of CMR in diagnosing flow limiting CAD were 100%, 70%, 81% and 100% respectively.In patients who underwent both stress CMR and nuclear stress test (n=20), stress CMR showed late gadolinium enhancement (LGE) in the coronary artery distribution in thirteen patients (n=13 of 20), consistent with fibrosis due to myocardial infarction. Where as nuclear stress test showed fixed defect in a coronary artery distribution in only six patients (n=6 of 20).Stress CMR also demonstrated other etiologies like hypertrophic cardiomyopathy, cardiac iron overload and idiopathic dilated cardiomyopathy.Stress CMR imaging has high sensitivity and negative predictive value in the identification of CAD, in patients with complete LBBB. It can also assess infiltrative cardiomyopathies. Stress CMR appears to provide a better comprehensive assessment than nuclear scintigraphy in patients with LBBB.None."} +{"text": "The astrocytoma cancer represents CNS neoplasms in which the predominant cell type is derived from an immortalized astrocyte. The genomewide analysis of glioma identified somatic mutation at codon 132 of the IDH1 gene which encodes NADP+ dependent isocitrate dehydrogenase. Further studies indicated that patients with somatic, heterozygous R132H mutation have distinct clinical characteristic: younger age at astrocytoma diagnosis (WHO II and WHO III) and improved clinical prognosis. Location of the majority of point mutations in the IDH1 gene are localized at 132 codon - what simplifies the use of this mutation for potential diagnostic purposes.The presence of R132H IDH1 mutation was analysed in group of 38 patients diagnosed with: fibrillar astrocytoma, astrocytoma gemistocyticum, astrocytoma pilocyticum and astrocytoma anaplasticum. The IDH1 mutation status was determinated by immunohistochemistry using monoclonal antibody specific for the R132H mutation. Additional data verification was performed by HRM Cold-PCR and Sanger sequencing. For statistical evaluation we distinguished two subgroups of patients: with and without IDH1 R132H mutation. Presence of IDH1 mutation in Polish astrocytomas\u2019 patients correlates with better clinical outcome and longer median overall survival. Our findings confirm overall tendency for better survival benefits in patients with IDH1 mutated tumors and indicates that presence or absence of IDH1 mutant proteins may become a potential target in personalized medicine."} +{"text": "Batrachochytrium dendrobatidis, [Bd]) identified three highly divergent genetic lineages, only one of which has a global distribution. Bd strains within these linages show variable genomic content due to differential loss of heterozygosity and recombination. The current quantitative polymerase chain reaction (qPCR) protocol to detect the fungus from amphibian skin swabs targets the intergenic transcribed spacer 1 (ITS1) region using a TaqMan fluorescent probe specific to Bd. We investigated the consequences of genomic differences in the quantification of ITS1 from eight distinct Bd strains, including representatives from North America, South America, the Caribbean, and Australia. To test for potential differences in amplification, we compared qPCR standards made from Bd zoospore counts for each strain, and showed that they differ significantly in amplification rates. To test potential mechanisms leading to strain differences in qPCR reaction parameters (slope and y-intercept), we: a) compared standard curves from the same strains made from extracted Bd genomic DNA in equimolar solutions, b) quantified the number of ITS1 copies per zoospore using a standard curve made from PCR-amplicons of the ITS1 region, and c) cloned and sequenced PCR-amplified ITS1 regions from these same strains to verify the presence of the probe site in all haplotypes. We found high strain variability in ITS1 copy number, ranging from 10 to 144 copies per single zoospore. Our results indicate that genome size might explain strain differences in ITS1 copy number, but not ITS1 sequence variation because the probe-binding site and primers were conserved across all haplotypes. For standards constructed from uncharacterized Bd strains, we recommend the use of single ITS1 PCR-amplicons as the absolute standard in conjunction with current quantitative assays to inform on copy number variation and provide universal estimates of pathogen zoospore loads from field-caught amphibians.Genomic studies of the amphibian-killing fungus ( Batrachochytrium dendrobatidis [Bd]) use qPCR to detect the pathogen on the skin of wild amphibian populations, providing a non-invasive sampling method that can yield diagnosis within a few hours Bd detection via qPCR has allowed researchers to detect infection levels in natural populations at different stages of emerging epidemics Bd epizootic waves spreading through na\u00efve populations Advances in quantitative polymerase chain reaction (qPCR) protocols and their application in detection and quantification of pathogens have contributed significantly to our understanding of disease dynamics in natural host populations Bd strains indicates that Bd is composed of at least three divergent genetic lineages that differ in virulence Bd strain recently discovered in Brazil differs in DNA content compared to GPL strains from Panama and California Recent genomic characterization of 20 global Bd via qPCR, researchers count zoospores from cultured Bd strains, extract genomic DNA (gDNA), and serially dilute to the desired concentrations genomic DNA (zoospore counts), 2) equimolar DNA solutions, and 3) ITS1 PCR amplicons. For each strain template, we tested differences in cycle threshold (Ct), defined as the point on the amplification curve associated with exponential growth of PCR product. We then used ITS1 PCR amplicons as a standard to quantify the ITS1 copy number from our focal strains. Finally, we cloned and Sanger-sequenced ITS1 PCR amplicons to compare ITS1 haplotype diversity among strains, which could lead to differences in amplification rates. Our study highlights the importance of understanding the evolutionary history of Bd at each sampling locality and the caveats of using genomic DNA as a standard for qPCR. We include a step-by-step protocol (Supporting Information) so Bd researchers can measure ITS1 copy numbers from any uncharacterized Bd strain and estimate infection intensity from field-collected amphibian skin swabs.In this study, we quantified and characterized ITS1 regions in multiple Bd strains isolated from amphibian hosts in different countries to generate replicate sets of DNA standards for qPCR: JEL404 , MexMkt (Mexico), JEL427 , PAB01 , LFT001_01 , CLFT023 , CLFT024 , and LBabercrom . Most of these Bd strains belong to the global panzootic lineage to others cultured from different host species across the New World . Because Bd quantification methods were developed using Australian strains We used Bd strains on 1% mTGh agar with penicillin-G and streptomycin sulfate 7 zoospores mL\u22121 for each strain. We then centrifuged all the aliquots at high speed for five minutes to obtain a zoospore pellet. We removed the supernatant and extracted the DNA from the pellet using 200 \u00b5L Prepman Ultra following Boyle et al. We grew Bd strain preparations to compare their efficiency as absolute quantification standards for several tandards . qPCR pe7 zoospores mL\u22121 three times using a Qubit\u00ae. For Standard Set A, we used a 5-log dynamic range starting at 103 zoospore genomic equivalents in 5 \u00b5L , we assumed 100% extraction efficiency and serially diluted to the required concentrations following Boyle et al. in 5 \u00b5L . For Sta size \u22121 .E. coli competent cells. To verify that bacterial cells incorporated the insert, we used blue-white screening, amplified the plasmid using M13 primers, and visualized the products on a 1.75% agarose gel. We cleaned the PCR products with ExoSap and quantified them with the Qubit\u00ae to estimate the mass of a fragment containing one copy of the ITS1/5.8S region. We estimated that this fragment weighs \u223c4.10\u00d710\u221210 ng, based on the total length of the M13 PCR-amplicon fragment and the average weight of one base pair (660 Da). Then, we built our third serial dilution and in 1.5 ng of DNA (Standard Set B).We performed endpoint PCR on strain JEL427 (Puerto Rico) using primers ITS1-3 Chytr and 5.8S Chytr a priori, rather we included Standard Set C in all qPCR runs and measured the cycle threshold (Ct) value for each of the dilution curves in Standard Sets A and B. We considered two threshold values in our analyses, the automatic threshold set by the ViiA7 software and a custom threshold of 0.1 to standardize among different runs.We modified the qPCR protocol in Boyle et al. (2004) by adding BSA To assess ITS haplotype diversity among strains, we cleaned 400-bp PCR amplicons (M13+ ITS1/5.8S region) using ExoSAP and sequenced 21\u201343 clones per strain using Big Dye v3.1 chemistry on an ABI 3730 sequencer . We aligned and edited chromatograms using Sequencher (Gene Codes Corp.), identified all unique haplotypes, and calculated haplotype frequency [defined as the number of sequences of particular haplotype over the total number of sequences for each strain]. We also verified the presence of the probe-binding site in each haplotype sequence.unpublished) for strains MexMkt (Mexico), JEL427 (Puerto Rico), LFT001_01 (Brazil), CLFT023 (Brazil), CLFT024 (Brazil), and LBAbercrom to assess whether the ITS1 haplotypes detected through cloning occurred in similar frequencies as in the genomic data. To estimate ITS1 haplotype frequencies in Bd genomes, we counted the number of times that each haplotype sequence recovered through cloning to determine which pairwise strain comparisons were statistically different. To quantify the number of ITS1/5.8S regions in each strain, we used a standard curve based on the ITS1 PCR-amplicon (Standard Set C) against each of the zoospore-based serial dilutions (Standard Set A and Standard Set B). We performed all statistical analyses using R Bd strain using STF statistics implemented in Arlequin 3.5 We compared the number of ITS1 haplotypes recovered by cloning and whole genome sequencing using a paired t-test. Finally, we calculated ITS1 haplotype frequencies for both data sets and estimated pairwise genetic differentiation among the ITS1 haplotypes for each t and a marginally lower slope in JEL404 (USA) . Therefore, most strains had similar amplification efficiencies (>98%) with the exception of PAB01 (Puerto Rico) and JEL404 (USA) which showed slightly higher efficiencies . All staciencies .7) extracted from each Bd strain had different DNA concentrations, that ranged from 1.01 to 4.11 ng \u00b5L\u22121 based on Qubit\u00ae quantification showed a significant negative relationship between nanograms of DNA and Ct . Our post-hoc Tukey's HSD tests revealed further pairwise similarities between strains in amplification profiles: JEL404-JEL427, LFT001_01-JEL427, LFT001_01-JEL404, and PAB01-MexMkt did not significantly differ from each other and holding zoospore quantity or input DNA constant (3), our results showed high copy number variability, which ranged from 10 copies in MexMkt to 144 copies in CLFT023 . This result indicates that cloning did not bias our relative estimates of haplotype diversity but failed to detect rare haplotypes. Strains MexMkt and LFT001_01 had different haplotype frequency distributions compared to JEL427, CLFT023, CLFT024, and LBabercrom to establish qPCR standard curves can be problematic because y number . These sBd qPCR standards made from strains collected in six countries points to patterns of genomic change during the evolution and global spread of BdBd strain in 1.5 ng of extracted DNA , Panama (JEL423), and Brazil also indicated significant variation in DNA content Bd strains JEL423 and JAM81 have different assembly sizes, which may result from the presence of chromosomal length polymorphisms Bd, and indicate that insertions and/or deletions of regions including ribosomal ITS1 may explain the differences we observed in qPCR standard curves across strains.All other strains showed similar slopes and intercepts for zoospore-based standard curves are often associated with disease epidemics, mortality, and population extirpations Bd standards.ITS1 copy number variation among Bd strains that have a 10-fold difference in the number of ITS1 regions. Furthermore, the possibility of sampling amphibians with co-infections of multiple strains with variable ITS1 copy number will remain a problem in wild populations, unless we characterize ITS1 regions and develop strain-specific assays. To reduce the potential biases caused by Bd genome variation or co-infections, we recommend that researchers combine PCR amplicon standards in conjunction with standards based on zoospore counts for detecting BdBd genomic equivalents. PCR-amplicon standards can be generated cheaply and will allow researchers to determine the number of ITS1 regions in newly isolated Bd strains. Our strain-independent method (see Supporting Information) complements traditional standard curves, and provides accurate and comparable measures of infection intensities across sampling sites and studies.10,000 zoospores and 1,000 zoospores can generate the same amount of fluorescence in qPCR assays if standards are made with Figure S1Bd strains. Light blue area represents unique 35 bp fragment used for searches in the Illumina sequence data. See for FASTA formatted sequences.An alignment of 26 ITS1 haplotypes detected by cloning and sequencing PCR products generated using primers ITS1-3 Chytr and 5.8S Chytr on genetic material from seven (EPS)Click here for additional data file.Figure S2Estimated DNA content per zoospore from ITS copy number and the number of copies in 1.5 ng DNA.(EPS)Click here for additional data file.Table S1Percent identity and range in coverage for each strain at each primer binding site. NR represents the total number of filtered reads searched.(DOCX)Click here for additional data file.Table S2STF values. Upper diagonal values are from cloning/Sanger sequencing and lower diagonal values from Illumina sequencing. Significant STF values (P<0.05) are shown in bold.Pairwise (DOCX)Click here for additional data file.Appendix S1FASTA formatted ITS1 haplotypes with gaps included.(DOCX)Click here for additional data file."} +{"text": "There were multiple errors in the legend for Figure 2. The correct legend is: \"Transmissibility of MPXV was studied within the prairie dog MPXV model. For each study, eight cages with holes on one side were utilized to individually cross-house eight animals. One challenged animal was cross-housed with an uninfected animal from the time of exposure of the challenged animal. West African MPXV primary challenge prairie dogs (n=3) are shown in blue line. High dose Congo Basin MPXV primary challenged prairie dogs (n=4) are shown in green lines . Low dose Congo Basin MPXV primary challenged prairie dogs (n=2) are shown in orange lines (PD9049 died on day on day 21 as indicated by cross).\""} +{"text": "Preclinical and clinical studies report that \u03b2-blockers may be an interesting option to attenuate the deleterious effects of prolonged catecholamine exposure during septic shock. Nevertheless, there are concerns that \u03b2-blockers may have negative chronotropic and inotropic effects leading to inappropriately low cardiac output. The objective of the present study was therefore to elucidate whether a reduction in heart rate (HR) with esmolol may negatively affect microcirculation in patients with septic shock who remained tachycardic after hemodynamic optimization.After 36 hours of initial hemodynamic stabilization, 11 septic shock patients with HR >95 bpm and requiring norepinephrine (NE) to maintain mean arterial pressure (MAP) between 65 and 75 mmHg, despite adequate volume resuscitation, received a continuous esmolol infusion to maintain HR between 94 and 80 bpm. NE was titrated to achieve a MAP between 65 and 75 mmHg. Data from right heart catheterization and sidestream dark-field imaging were obtained at baseline and after 24 hours.P < 0.05), stroke volume (SV), microvascular flow index of the small vessels (MFIs) and norepinephrine requirements did not vary during the 24-hour observational period. Results are summarized in Table Apart from a statistically significant decrease in HR and cardiac index (CI) (In patients with established septic shock who remained tachycardic after hemodynamic optimization in accordance with the current guidelines, titration of esmolol to reduce the HR to a predefined threshold did not affect microcirculatory blood flow."} +{"text": "Activating mutations of the serine threonine kinase v-RAF murine sarcoma viral oncogene homologue B1 (BRAF), most commonly of the V600E type, are found in a wide range of human neoplasms including primary and secondary brain tumors. Therapeutic BRAF inhibitors have shown clinically meaningful activity, particularly in metastatic BRAF V600E mutated melanoma including patients with brain metastases. Therefore, in current neuropathological practice BRAF testing is of clinical importance in tissue samples of melanoma brain metastases in order to identify cases amenable to therapy with BRAF inhibitors. BRAF mutation testing may also add additional information for differential diagnosis of primary brain tumors in selected situations, e.g., for differentiation of anaplastic pleomorphic xanthoastrocytoma (BRAF V600E mutation in 65%) from glioblastoma (BRAF V600E mutation in < 5%). The BRAF mutation status can be tested with DNA-based methods and immunohistochemistry using a V600E mutation-specific antibody. In summary, at this point BRAF V600E testing is clinically indicated in relatively few cases of the daily clinical neuropathology practice, but has important predictive implications for patients with melanoma brain metastases. Depending on the results of additional clinical studies, determination of BRAF mutation status may become clinically relevant also for primary brain tumors such as glioblastoma in the future. Activating mutations of the serine threonine kinase v-RAF murine sarcoma viral oncogene homologue B1 (BRAF) are found in a wide range of human cancers and represent the most frequent unique genetic alteration among melanoma 50 \u2013 60%), papillary thyroid carcinoma (40 \u2013 70%) and hairy cell leukemia (> 90%) . Among p0 \u2013 60%, Testing for the presence of BRAF V600E mutation may be relevant in clinical neuropathology practice for the following reason: BRAF mutations affect some brain tumors (both primary and secondary) , 6 and sIn current neuropathological practice, BRAF testing can be of clinical value in tissue samples of melanoma brain metastases from patients with unknown mutation status in order to identify cases amenable to therapy with BRAF inhibitors. The BRAF status has been shown not to vary between different tumor manifestations of the same patient . Thus, iThe BRAF mutation status can be analyzed using DNA-based methods such as the US Food and Drug Administration (FDA) approved Cobas 4800 BRAF V600 test , which is commercially available. Recently, a monoclonal antibody (antibody VE1) which reliably detects BRAF V600E mutated protein in formalin-fixed and paraffin-embedded tumor tissue samples has been generated . VE1 antIn summary, at this point BRAF V600E testing is clinically indicated in relatively few cases of the daily clinical neuropathology practice, but has important therapeutic implications in selected patients. It seems advisable that implementation of the test should be a joint decision of (neuro-)pathologists and (neuro-)oncologists at a given center. Andreas von Deimling and David Capper declare shared inventorship of BRAF antibody clone VE1. A patent for diagnostic application of VE1 has been applied for. All terms are being managed by the German Cancer Research Center in accordance with its conflict of interest policies. Dr. Preusser has recieved lecture honoraria and travel and research support by Roche Austria."} +{"text": "Distributive shock with high output cardiac failure is frequently seen in acute liver failure (ALF). A previous study suggested a high incidence of myocardial injury coupled with adverse outcome in this population . CorrelaNT-proBNP (NTpBNP) and troponin I (TI) were measured in ALF patients with shock within 48 hours after admission to a tertiary specialist ICU. Transpulmonary thermodilution cardiac output monitoring (PiCCO) was performed in all patients. Values of cardiac index (CI), stroke volume index (SVI), global end diastolic index (GEDI) and markers of contractility - global ejection fraction (GEF) and cardiac function index (CFI) - as well as severity of illness scores were correlated with cardiac biomarker levels. Correlation was assessed using Pearson's coefficient for normally distributed data.P = 0.001) and TI but not with PiCCO parameters related to flow, contractility or preload and neither cardiac marker correlated with APACHE II score. There was a trend toward correlation of TI with CFI but not with CI . CFI was correlated with GEF and lactate . APACHE and SOFA did not correlate significantly with PiCCO indices.Twenty-six ALF patients with a mean (SD) APACHE II score of 23 (4) and SOFA 15 (2) were assessed. NTpBNP pg/ml) and TI (median 0.28 (0 to 50) u/l) levels were both significantly elevated without any significant ECHO abnormalities and 24 patients required renal replacement therapy. Serum NTpBNP correlated with serum lactate (correlation coefficient 0.61, Levels of cardiac biomarkers are frequently elevated in ALF. We could not find any correlation of TI and NTpBNP with surrogate markers of cardiac function on invasive hemodynamic monitoring, or indeed significant abnormalities on ECHO."} +{"text": "Right ventricular (RV) function predicts prognosis in pulmonary hypertension (PH) patients (pts) and Right Ventricular failure (RV). Prior studies evaluating of 3D RV ejection fraction (EF) have yielded inconsistent prognostic information. Here we explore the prognostic value of contrast induced CMR in PH patients with RV dysfunction.We hypothesize that myocardial Late Gadolinium Enhancement (LGE) which a marker for myocardial fibrosis when present in RV or RV insertion points (RVIP) is a predictor of poor prognosis in PH pts.A retrospective chart review of PH pts (n=21) who underwent clinically indicated CMR were analyzed. Demographic data showed mean age 61 yrs; 28% male; 43 % WHO group I, 29% group II, 14% group IV,14% group V. RV volumetric data were indexed to BSA, and along with RVIP LGE information were correlated with major adverse clinical events (MACE) such as hospitalization, death and referral/need for lung transplantation.LGE was positive (+) in 14 pts (66%) and (-) in the remaining 7 pts (33%). Compared to LGE (- ) pts, the LGE (+) pts had higher RVEDVI, RVESVI and lower RVEF (p<0.05). However, only LGE in combination with 3D RVEF predicted MACE. Specifically, LGE (+) pts had 9 MCE compared to 1 in the LGE (-) pts. The median RVEF value was 45%. The event rate was 89% in pts with LGE (+) and RVEF<45 compared to 20% in LGE (-) and RVEF <45%. Group comparisons were done using the Fisher\u2019s exact test. The comparisons were not statistically significant due to small sample size.Late Gadolinium Enhancement may be the pathophysiologic hallmark in patients with PH patients as a direct reflection of the underlying RV failure due to progressive myocardial fibrosis. Late gadolinium enhancement in combination with RVEF provides important prognostic information that accurately predicts important adverse CV outcomes such as lung transplantation and death. Prospective studies are needed to confirm this observation."} +{"text": "STAT3 inhibition was also effective in treating an RA model, collagen induced arthritis (CIA), in vivo through significant reduction in expression of inflammatory cytokines and RANKL, inhibiting both inflammation and joint destruction. Thus our data provide new insight into pathogenesis of RA and provide evidence that inflammatory cytokines induce a cytokine amplification loop via STAT3 that promotes sustained inflammation and joint destruction.Rheumatoid arthritis (RA) causes sever joint damage and significant disability of daily living. The symptoms of RA patients are mainly from chronic inflammation and continuous joint destruction, however, the mechanisms underlying how inflammation and joint destruction in RA develop and are sustained chronically remain largely unclear. In this study, we show that signal transducer and activator of transcription 3 (STAT3) plays a critical role in both chronic inflammation and joint destruction in RA. We found that inflammatory cytokines, such as IL-1\u03b2, TNF\u03b1 and IL-6, activated STAT3 either directly or indirectly and induced expression of inflammatory cytokines, further activating STAT3. STAT3 activation also induced expression of receptor activator of nuclear factor kappa B ligand (RANKL), an essential cytokine for osteoclast differentiation. STAT3 knockout or pharmacological inhibition resulted in significant reduction of the expression of both inflammatory cytokines and RANKL"} +{"text": "LRRC6 (Leucine-rich repeat containing protein 6) that fit the model of autosomal recessive genetic transmission, leading to a change of a highly conserved amino acid from aspartic acid to histidine (Asp146His). LRRC6 was localized to the cytoplasm and was up-regulated during ciliogenesis in human airway epithelial cells in a Foxj1-dependent fashion. Nasal epithelial cells isolated from affected individuals and shRNA-mediated silencing in human airway epithelial cells, showed reduced LRRC6 expression, absent dynein arms, and slowed cilia beat frequency. Dynein arm proteins were either absent or mislocalized to the cytoplasm in airway epithelial cells from a primary ciliary dyskinesia subject. These findings suggest that LRRC6 plays a role in dynein arm assembly or trafficking and when mutated leads to primary ciliary dyskinesia with laterality defects.Despite recent progress in defining the ciliome, the genetic basis for many cases of primary ciliary dyskinesia (PCD) remains elusive. We evaluated five children from two unrelated, consanguineous Palestinian families who had PCD with typical clinical features, reduced nasal nitric oxide concentrations, and absent dynein arms. Linkage analyses revealed a single common homozygous region on chromosome 8 and one candidate was conserved in organisms with motile cilia. Sequencing revealed a single novel mutation in A motile cilium is composed of an axoneme containing nine outer microtubule doublets and an inner central pair. The outer doublets are associated with dynein motor proteins, organized as outer dynein arms (ODA) and inner dynein arms (IDA). These proteins allow adjacent outer doublets to slide against one other and thus provide movement. Nexin links tether and limit the motion of microtubular doublets, and radial spokes control dynein arm activity relaying signals from the central microtubular pair to the dynein arms Chlamydomonas reinhardtii) to zebrafish (Danio rerio) to mammals, have provided insights into structure, function, and genetics of the human cilium. Thus far, studies using these organisms and others have led to the identification of sixteen different genes that when mutated produce unambiguous clinical phenotypes of PCD in humans. These genes include DNAH5 (MIM 603335), DNAI1 (MIM 604366), DNAI2 (MIM 605483), TXNDC3 (MIM 607421), DNAL1 (MIM 610062), DNAH11 (MIM 603339), HEATR2 (MIM 614864), DNAAF1 (MIM 612517), DNAAF2 (MIM 613190), DNAAF3 (MIM 614566), RSPH4A (MIM 612647), RSPH9 (MIM 612648), CCDC39 (MIM 613798), CCDC40 (MIM 613799), CCDC103 (MIM 614677) and HYDIN (MIM 610812 ) DNAAF1, DNAAF2, DNAAF3 and HEATR2, encode proteins that are involved in dynein arm assembly while the others are essential structural components of the ciliary axoneme. Nonetheless, mutations in these genes still account for less than half of all PCD cases, and our understanding of the critical components of cilia assembly is incomplete Our understanding of the link between genetic defects and ultrastructural changes of cilia has greatly advanced over the past decade. Owing to conservation of cilia and flagellar structures, studies of these organelles in model organisms, from algae , flies (Drosophila melanogaster), protozoa (Trypanosoma brucei), algae (C. reinhardtii), but not in worms (Caenorhabditis elegans). There are fourteen other proteins with leucine-rich repeats (LRR) in the cilia proteome LRRC6 in normal airway epithelial cells.Here, we describe a single non-synonymous mutation in and Subjects with clinical features consistent with PCD from two unrelated, endogamous families were studied and Table 1All individuals or their parents provided written informed consent for diagnostic evaluation and genetic characterization. The study protocol was approved by the Hadassah-Hebrew University Human Subjects Committee. Institutional approval was obtained to conduct both human and animal research. Anonymized human airway epithelial cells from surgical excess of large airways that were trimmed during the transplant procedure, of lung donated for transplantation at Washington University in St. Louis were also used in these studies. Research using cells originating from deidentified cadaver specimen is exempt from regulation and is not governed by NIH regulation 45 CFR Part 46.Genetic linkage analysis was performed on three affected members , GAACACAACGACTGT GTCATT (shRNA#2), GATCTCAGACAACGGGTCATT (shRNA#3), CCTGTTTGTTTACTCCT GAAT (shRNA#4) and CCTAAATGTGAATGAGCCCAA (shRNA#5). A non-targeted sequence with a yellow fluorescent protein (YFP) reporter (a gift from Y. Feng and G.D. Longmore), was used as control shRNA targeted sequences generated by the Children\u2019s Discovery Institute shRNA Library Core, were inserted into pLKO.1 lentivirus vectors that includes a U3 promoter and a puromycin resistant cassette. The shRNA sequences used were: Normal human lung obtained from excess tissue donated for lung transplantation was fixed, immunostained and imaged as previously described LRRC6, 5\u2032-GCAGGCTTTGATGGACGTTG and 5\u2032-GCCTGTAGGTGGTCTTTGCT; murine LRRC6, 5\u2032-AAGTTGACCCCAGCAAGCAT and 5\u2032-CTCACTGGGTTCATCTCGGG; Foxj1, 5\u2032-CCCGACGACGTGGACTAC and 5\u2032-GGCGGAA GTAGCAGAAGTTG; DNAI1, 5\u2032-AACGACGGCTGTCCCTAAAG and 5\u2032-AGCCTACAAAACGC TCCCTC; and DNAH7, 5\u2032-ACTTGCAGAATCGCATCCCA and 5\u2032-CTCCTCTCCGCTC ACTTGTC, and detected using SYBR green in Lightcycler 480 RNA expression was assessed by RT-PCR amplification using the following oligonucleotide primer sets: human Nasal epithelial cells collected from subjects with PCD were examined using previously published protocols Data are expressed as mean \u00b1 standard deviation (SD). Statistical comparisons between groups were made using single factor analysis of variance (ANOVA) with Bonferroni correction for multiple comparisons. Individual comparisons were made using Student Two-tail test.andSupplementary S2) and absent dynein arms were found in at least one of the affected siblings from each family. Analysis of single-nucleotide polymorphism (SNP) haplotype on three affected members in one family , EIF2C2 (MIM 606229), NDRG1 (MIM 605262), EFR3A (MIM 611798), EIF2C2 (606229), and DDEF1 (MIM 605953)] were annotated in the ciliary proteome LRRC6 (leucine-rich repeat containing protein 6), which resulted in substitution of aspartic acid to histidine (Asp146His) , Seattle, WA).Five subjects with clinical features consistent with PCD, including chronic sinusitis, bronchiectasis, recurrent otitis media and laterality defects, from two unrelated, consanguineous Palestinian families were studied andTable 1LRRC6 was originally identified as LRTP and expressed during spermatogenesis in mice and humans C. reinhardtii orthologue was increased following deflagellation when compared to pretreatment values, consistent with transcriptional up-regulation of flagellar genes during cillogenesis D. rerio (Lrrc6l), when mutated, results in ciliary motility defects ranging from immotility to disorganized beating in the pronephros and neural tube, but normal axonemal ultrastructure D. melanogaster tilB mutants have defective sperm flagella motility and dysfunctional ciliated dendrites of the chordotonal organs. Furthermore, these mutant sperm axoneme lacked dynein arms T. brucei localizes to basal bodies and is critical for basal body duplication, flagellum assembly, and cytokinesis Supplementary Figure S1), suggesting its involvement in assembly or trafficking during cilia biogenesis.To better elucidate the function of LRRC6 in cilia assembly, we examined its expression in human airway epithelial cells. LRRC6 was not found in the ciliary axoneme, but was distributed throughout the cytoplasm of ciliated airway epithelial cells . Cilia were present on the apical surface of cells treated with shRNA sequences and affected individuals, which showed that LRRC6 was not required for ciliogenesis . When compared to non-PCD or non-targeted shRNA transfected cells .) suggesting mislocalization of the protein and failure of axonemal transport. In contrast, DNAH7 was not detected in the PCD cells, which may be related to protein degradation or suppressed expression. The latter was further evaluated by examining the expression of DNAI1 and DNAH7 in nasal cells for PCD subjects. DNAI1 and DNAH7 transcription was markedly reduced in nasal cells from three PCD subjects compared to a healthy control; findings that were also recapitulated in Lrrc6-specific shRNA targeted cells, suggesting that mutations in LRRC6 alters the expression of genes encoding some ODA and IDA proteins To examine the role of LRRC6 in dynein arm assembly, we immunostained ciliated cells with antibodies against DNAI1, an outer dynein arm polypeptide, and DNAH7, an inner dynein arm polypeptide. Neither DNAI1 nor DNAH7 were detected in cilia from PCD subjects, but DNAI1 was found in the apical cytoplasm of the epithelial cell and Foxj1-deficient (\u2212/\u2212Foxj1) mice Lrrc6 levels were markedly reduced in \u2212/\u2212Foxj1 airway epithelial cells when compared to +/+Foxj1 cells with markers of endosomes (green), and lysosomes (green). However, LRRC6 localized with \u03c7-tubulin, a marker for basal bodies (green). Nuclei were stained using DAPI (blue). acetylated \u03b1-tubulin, a cilia marker, is shown in turquoise .(TIF)Click here for additional data file.Figure S2RT PCR analysis of LRRC6 expression in RNAi silenced cells.(A)LRRC6 expression in LRRC6-specific shRNA transfected airway epithelial cells (B) Immunoblot analyses of airway epithelial cells transfected with three different LRRC6-specific shRNA or non-targeted shRNA (NT) sequences and nontransfected control cells (M). (C) En face images of LRRC6 in cultured preparations of ciliated airway epithelial cells from a normal donor, transfected with either non-targeted, control shRNA (NT) or different LRRC6 targeted shRNA sequences. LRRC6 (red), acetylated \u03b1-tubulin (green), a ciliated cell marker, and co-stained with DAPI (blue). .(TIF)Click here for additional data file.Video S1Healthy human nasal epithelial cells. Nasal epithelial cells from a healthy non-PCD subject showing normal cilia motion.(MP4)Click here for additional data file.Video S2Nasal epithelial cells from a subject with PCD. Nasal epithelial cells from a PCD subject with the LRRC6 mutation showing no cilia motion.(MP4)Click here for additional data file."} +{"text": "This study evaluated the potential adverse effects associated with exposure to calcium carbonate precipitate during continuous venovenous hemofiltration (CVVH). The clinical use of Accusol 35 Solution (Accusol 35) has been associated with occasional formation of calcium carbonate precipitate in the tubing set during therapy.n = 6) or negative control article (n = 8) for 6 hours. The test article was Accusol 35 with induced precipitate formation prior to CVVH. The test article contained visible particles and subvisible particles 36\u00d7 higher than the maximum concentration specified in the European Pharmacopoeia (EP). The negative control article was Accusol 35 containing no visible particles and subvisible particles within EP specification. One-half of the dogs in the negative control article group received a central venous injection of Sephadex G-50 beads (10 mg/kg) following CVVH as a positive control. Select cardiovascular (CV) parameters were monitored continuously or were calculated at predetermined times. Arterial samples were obtained at predetermined times for analysis of blood gases and electrolytes. Samples of the test and negative control articles were obtained hourly during CVVH for determination of pH and subvisible particles. Dogs were euthanized and lung tissue samples were examined histologically.Fourteen mongrel dogs were anesthetized, instrumented, and received CVVH with the test (P < 0.01) in mean pulmonary arterial pressure due solely to a similar increase (P < 0.01) in pulmonary vascular resistance. No differences in blood gases or electrolytes were observed between the test and negative control articles. Sephadex beads caused a decrease (P > 0.05) in arterial blood PO2 and an increase (P > 0.05) in arterial blood PCO2. No differences in lung histology were observed between the test and negative control articles. The lungs from all dogs given Sephadex beads contained multiple intravascular particles in large-caliber blood vessels.All CV parameters remained stable and no differences were observed between the test and negative control articles. Sephadex beads caused an increase (CVVH performed on anesthetized dogs for 6 hours using Accusol 35 containing visible and subvisible particles 36\u00d7 higher than the maximum concentration specified in the EP resulted in no adverse effects on CV parameters, blood gases and electrolytes, and lung histology as compared with Accusol 35 containing no visible particles and subvisible particles that were within EP specification."} +{"text": "CMR is an accurate and reproducible technique for the analysis of left ventricular volumes in adults but less well validated in children. Various analysis tools and segmentation methods are available but it is not clear which is most appropriate for use in children. Conventional manual segmentation tools require time-consuming contour tracing but allow some compensation for image mis-registration. The semi-automated tool employs a heart model based algorithm with reported significantly reduced analysis times but correction for the image mis-registration more frequently seen in children is more difficult.To compare manual and semi-automated analysis tools used for the assessment of left ventricular (LV) volumes and mass measured using cardiovascular magnetic resonance (CMR).CMR was performed in 10 healthy children aged 9 years as part of a study of developmental influences on cardiovascular structure and function. Contiguous short axis steady state free precession LV cine images were acquired. Scans were repeated following a short interval. Data sets were analyzed to calculate LV volumes and mass using a manual technique (Osirix) and a semi-automated technique (Argus 4D). Papillary muscles and trabeculae were included in the blood pool. LV stroke volumes (SV) from each technique were compared with aortic flow data derived from aortic valve phase contrast velocity flow mapping sequences .Using the Bland Altman method, the mean difference in SVs for Osirix and Argus 4D was 7ml, with Argus 4D generally measuring larger values. The estimated coefficient of variation for SV measurements calculated using Osirix was lower than that using Argus 4D (10.6% vs 13.1%). Mean differences between the Osirix and Argus 4D SVs and aortic flow were 1.4ml and 5.7ml, respectively.In children SV measurements tend to be more reproducible using an Osirix manual technique compared with a semi-automated tool (Argus 4D). Osirix derived SVs were more accurate than Argus 4D when compared with aortic phase contrast derived flow data."} +{"text": "Combined pelvic floor electromyography (EMG) and videocystourethrography (VCUG) during urodynamic investigation are the most acceptable and widely agreed methods for diagnosing detrusor external sphincter dyssynergia (DESD). Theoretically, external urethral sphincter pressure (EUSP) measurement would provide enough information for the diagnosis of DESD and could simplify the urodynamic investigation replacing combined pelvic floor EMG and VCUG. Thus, we evaluated the diagnostic accuracy of EUSP measurement for DESD.A consecutive series of 72 patients with neurogenic lower urinary tract dysfunction able to void spontaneously was prospectively evaluated at a single university spinal cord injury center. Diagnosis of DESD using EUSP measurement (index test) versus combined pelvic floor EMG and VCUG (reference standard) was assessed according to the recommendations of the Standards for Reporting of Diagnostic Accuracy Initiative.Using EUSP measurement (index test) and combined pelvic floor EMG and VCUR (reference standard), DESD was diagnosed in 10 (14%) and in 41 (57%) patients, respectively. More than half of the patients presented discordant diagnosis between the index test and the reference standard. Among 41 patients with DESD diagnosed by combined pelvic floor EMG and VCUR, EUSP measurement identified only 6 patients. EUSP measurement had a sensitivity of 15% (95% CI 5%\u201325%), specificity of 87% (95% CI 76%\u201398%), positive predictive value of 60% (95% CI 30%\u201390%), and negative predictive value of 56% (95% CI 44%\u201368%) for the diagnosis of DESD.For diagnosis of DESD, EUSP measurement is inaccurate and cannot replace combined pelvic floor EMG and VCUR. Detrusor external sphincter dyssynergia (DESD) is defined as a detrusor contraction concurrent with an involuntary contraction of the urethral and/or periurethral striated muscle Despite the high clinical relevance of DESD, there is no single \u201cgold standard\u201d method for its diagnosis. Blaivas and Fisher We hypothesized that the measurement of external urethral sphincter pressure (EUSP) could replace combined pelvic floor EMG and VCUR for the diagnosis of DESD. Thus, we prospectively evaluated the diagnostic accuracy of EUSP measurement for DESD.This study was approved by the local ethics committee of the University of Z\u00fcrich and registered with ClinicalTrials.gov (study registration number: NCT01293110). All participants gave written informed consent.From November 2010 to April 2011, 191 consecutive patients older than 18 years with neurogenic lower urinary tract dysfunction (NLUTD) underwent video-urodynamic investigation at the Spinal Cord Injury Center, Balgrist University Hospital, Z\u00fcrich, Switzerland. Of those, 76 (39%) could void spontaneously and were prospectively enrolled into the study.All methods, definitions, and units are according to the standards recommended by the International Continence Society Video-urodynamic investigations were performed according to Good Urodynamic Practices recommended by the International Continence Society Blood pressure and heart rate were measured at the beginning and the end of the video-urodynamic investigation which was interrupted immediately in the case of signs of autonomic dysreflexia.2O of EUSP during the voiding phase. During video-urodynamic investigations, EUSP measurement (index test) and combined pelvic floor EMG and VCUG (reference standard) were performed simultaneously.According to the literature All video-urodynamic investigations were assessed by two experienced urologists in consensus. Pelvic floor EMG and VCUG were interpreted blinded to the EUSP measurements and vice versa.The outcome measure was the diagnosis of DESD using EUSP measurement (index test) versus combined pelvic floor EMG and VCUG (reference standard).Data were normally distributed and they are presented as mean \u00b1 standard deviation (SD). Sensitivity, specificity, and predictive values of EUSP measurements including 95% confidence intervals (CI) were calculated. Statistical analyses were performed using SPSS version 17.0 .Of the 76 eligible patients, 4 with unclear results due to technical problems were excluded leaving 72 patients for analysis. Thirty six (50%) were women and 36 (50%) men. Mean age was 52\u00b114 years (range 19\u201380). The causes of NLUTD were spinal cord injury in 40 (55%), multiple sclerosis in 10 (14%), spinal stenosis in 4 (6%), cauda equina syndrome in 2 (3%), spina bifida in 2 (3%), disk hernia in 2 (3%), and others in 12 (16%).All patients voided spontaneously and as an adjunct 9 (12%) relied on clean intermittent self-catheterization and 8 (11%) on an indwelling suprapubic catheter. Fifty one (71%) patients had no medication for lower urinary tract dysfunction, 15 (21%) were on antimuscarinics, and 7 (10%) on alpha-blockers.Urodynamic findings are summarized in Using EUSP measurement (index test) and combined pelvic floor EMG and VCUR (reference standard), DESD was diagnosed in 10 (14%) and in 41 (57%) patients, respectively . More thNo adverse events related to the EUSP measurement (index test) and combined pelvic floor EMG and VCUR (reference standard) occurred during the study.In the present study including 72 patients with NLUTD, EUSP measurements were inaccurate for the diagnosis of DESD. Thus, in contrast to our initial hypothesis, EUSP measurement cannot replace combined pelvic floor EMG and VCUR and is therefore not recommended to assess DESD.2O in women, 53\u00b147 cmH2O in men, and >25 cmH2O in the majority of the patients. On the other side, among the patients who presented VCUR demonstrating incomplete/absent urethral opening at the external urethral sphincter site or increased/persistent EMG activity during the voiding phase, the mean reduction of EUSP was significantly smaller (6.4\u00b16.7 cmH2O in women and 5.0\u00b19.5 cmH2O in men). We therefore hypothesized that EUSP measurement could be an accurate method for the diagnosis of DESD.Sakakibara et al. Although the clinical and pathophysiological definition of DESD is standardized by the International Continence Society With the advent of multiple transducer catheters, simultaneous measurement of urethral and intravesical pressure using the same catheter has become possible. This seems a promising method for the diagnosis of DESD since the urodynamic investigation may be relevantly shortened and simplified. In addition, replacing combined pelvic floor EMG and VCUR by EUSP for the diagnosis of DESD would reduce the investigative costs and also the radiation exposure during urodynamics. However, we found inacceptable accuracy of EUSP measurement for the diagnosis of DESD with a false negative rate of 85% and low positive and negative predictive values not supporting the use of this technique in daily clinical practice.2O. Further research involving basic science and engineering technology is necessary in order to improve urethral pressure measurements.It is generally agreed that urethral pressure is of significant value for lower urinary tract function To the best of our knowledge, this is the first study investigating the accuracy of EUSP measurement for the diagnosis of DESD. Although our study complies with the recommendations of the Standards for Reporting of Diagnostic Accuracy Initiative For the diagnosis of DESD using EUSP measurement, we found sensitivity, specificity, positive and negative predictive values of 15%, 87%, 60%, and 56%, respectively. Thus, EUSP measurement is inaccurate and cannot replace combined pelvic floor EMG and VCUR to assess DESD.Figure S1Female sub-group analysis: Using EUSP measurement (index test) and combined pelvic floor EMG and VCUR (reference standard), DESD was diagnosed in 5 (14%) and in 23 (64%) female patients, respectively. More than 60% of the female patients presented discordant diagnosis between the index test and the reference standard. Among 23 female patients with DESD diagnosed by combined pelvic floor EMG and VCUR, EUSP measurement identified only 3 female patients. In females, EUSP measurement had a sensitivity of 13% (95% CI 4%\u201332%), specificity of 84% (95% CI 57%\u201395%), positive predictive value of 60% (95% CI 11%\u201396%), and negative predictive value of 35% (95% CI 19%\u201354%) for the diagnosis of DESD.(TIF)Click here for additional data file.Figure S2Male sub-group analysis: Using EUSP measurement (index test) and combined pelvic floor EMG and VCUR (reference standard), DESD was diagnosed in 5 (14%) and in 18 (50%) male patients, respectively. Almost half of the male patients presented discordant diagnosis between the index test and the reference standard. Among 18 male patients with DESD diagnosed by combined pelvic floor EMG and VCUR, EUSP measurement identified only 3 male patients. In males, EUSP measurement had a sensitivity of 16% (95% CI 5%\u201339%), specificity of 89% (95% CI 67%\u201397%), positive predictive value of 60% (95% CI 11%\u201396%), and negative predictive value of 51% (95% CI 33%\u201369%) for the diagnosis of DESD.(TIF)Click here for additional data file."} +{"text": "To the Editor: The influenza A pandemic (H1N1) 2009 virus rapidly created a global pandemic among humans and also appears to have strong infectivity for a broad range of animal species (\u2013\u2013Taxidea taxus taxus), a 19-year-old female Bornean binturong (Arctictis binturong penicillatus), and a 7-year-old black-footed ferret (Mustela nigripes) housed in a San Diego zoological garden.In 2009, the first recognized occurrence of pandemic (H1N1) 2009 in southern California in April was followed by a surge of cases during October through November 2009. Influenza A virus was not detected in samples from the ferret after it recovered. Results of PCRs for all other viruses were negative. Immunohistochemical evaluation of lung samples from the badger for antigens of influenza A virus 2009 virus. The badger and binturong were generally healthy, no other pathogens were detected, and pulmonary lesions were consistent with influenza pneumonia. In these animals, pandemic (H1N1) 2009 infection was especially aggressive, resulting in irreversible disease. Reports of pandemic (H1N1) 2009 virus in skunks and anteaters also describe severe disease in those species (Mustela putorius furo) than typical seasonal influenza viruses 2009 virus was more pathogenic in domestic ferrets 2009 virus may have a broader host range and stronger virulence than viruses in the past.Pandemic (H1N1) 2009 was first detected in humans in March 2009 and reached pandemic levels by June of that year, rapidly establishing a rich pool for the development of genetic variants. Naturally acquired disease has now been described in 10 animal species, and experimental infection has been reported in an additional 2 animals (mice and cynomolgus macaques) ("} +{"text": "Myocardial perfusion cardiac magnetic resonance imaging (MRI) is established as a high sensitive procedure to diagnose coronary artery disease (CAD). In practice exercise electrocardigram (XECG) is widely used as non invasive method to detect relevant coronary stenosis (>50%).The aim of this study was to investigate the diagnostic perfomance of adenosin stress 3-Tesla cardiac MRI for verification of ischaemia at coronary artery disease in comparison to XECG referred to coronary angiography as gold standard.We included patients who received XECG, stress cardiac MRI and accomplished invasive coronary diagnostics. The MRI measurements at 3.0 T based on cine-mode-sequences in short axes images, rest and adenosin stress (140 \u00b5g/kg bw/min) perfusion and late gadolinium enhancement imaging . Myocardial ischemia was defined as an area of perfusion deficit at stress MRI with negative late enhancement in areas of hypoperfusion (Panels A and B).XECG was carried out on bicycle ergometer with a standardised protocol.40 patients (63\u00b111 years) were analysed prospectively, ten patients were assumed to have CAD, 30 had known CAD and were supposed to have a progress. A myocardial infarction in history was known at 14 patients,XECG showed pathological findings in 24 cases (60%), twelve of these had angina or severe dyspnea, eleven patients had significant horizontal or down sloping ST-depression, one patient had an unsustained ventricular tachycardia.Coronary angiography investigated a significant CAD (stenosis \u226550%) at 30 patients.10 with normal XECG had relevant CAD. Sensitivity of XECG was 67%, specifity 60%. The positive predictive value (PPV) was calculated with 83%, the negative predictive value (NPV) was 37%.Adenosin stress cardiac MRI revealed perfusion deficits in 32 patients (80%). The sensitivity of MRI concerning coronary stenosis \u226550% was 93%, the specifity 60%. Two patients with inconspicuous adenosin stress cardiac MRI had significant CAD. The PPV of MRI was 88%, the NPV 75%. All 14 patients with prior myocardial infarction were detected by LGE-sequence (both sensitivity and specifity 100%).Myocardial perfusion magnetic resonance imaging at 3.0 Tesla is superior to exercise electrocardigram to diagnose relevant (stenosis \u226550%) coronary artery disease. Cardiac MRI could also demarcate all patients with prior myocardial infarction by late enhancement."} +{"text": "T1 mapping enables true quantitative assessment of myocardial tissue, separating scars and healthy myocardium. The modified Look-Locker inversion recovery (MOLLI) sequence acquires eleven images over the duration of a minimum of fifteen heart beats to obtain the final T1 map, which is susceptible to undesired respiratory motion due to a relatively long breath-hold. We aimed to implement a motion correction method using a hierarchical local affine registration of MOLLI images and tested if it improved image quality and thus produced more accurate post contrast myocardial tissue T1 estimation.Motion correction was implemented on T1 mapping images acquired 20 minutes post gadolinium contrast injection in 15 patients with a previous history of chronic myocardial infarction. Three experiments were carried out to compare the pre- and post-motion correction images. 1) Comparing visual quality score assessment of the T1 maps by two experienced independent observers; 2) comparing histogram distributions of voxel T1 values within the myocardium to allow tissue characterisation between the scarred region and the non-infarct region; and 3) comparing the \"error of mismatch\" measured by voxel misalignment in LV endocardial and epicardial borders. Wilcoxon signed rank test was performed to test for significant differences between the median errors of mismatch between the pre and post motion correction images.1) Visual qualitative assessment via two independent observers confirmed motion correction produced an improvement in image quality in 11 out of 15 patients. 2) The histograms assessing tissue characterisation showed an improvement in tissue T1 delineation after motion correction, demonstrated by more distinct signal intensity peaks after motion correction (Figure Motion correction using hierarchical local affine registration yields better imaging quality thus allowing more accurate tissue characterization from T1 maps acquired with MOLLI sequence.n/a"} +{"text": "Profound hemodynamic changes seen in acute liver failure (ALF) resemble those found in later stages of septic shock. Vasopressor support is frequently required and indiscriminate fluid resuscitation can worsen intracranial hypertension (ICH) and lung injury. Markers of preload dependency have thus far not been studied in this patient group and response to dynamic manoeuvres such as passive leg raising or end expiratory hold cannot be considered safe due to the high incidence of ICH.ALF patients admitted to a tertiary specialist ICU in vasoplegic shock, requiring multiorgan support including controlled mechanical ventilation, had their cardiac output monitored via transpulmonary thermodilution and pulse contour analysis (PiCCO). Markers of fluid responsiveness were compared between responders (CI \u226515%) and nonresponders to a colloid fluid challenge (5 ml/kg IBW). All patients had a transthoracic echocardiogram performed before and after fluid administration. The predictive capacity of stroke volume, pulse pressure variation and respiratory change in peak aortic velocity \u0394V peak for preload dependency was analyzed.R = 0.726, P < 0.001). There was no difference between those defined as responders using a cut-off value of CI or SVI of 10%. When using 15%, seven patients would have been classified differently. The intraclass correlation coefficient for CI and SVI change was 0.83 (0.62 to 0.92), confirmed using Pasing and Blakock regression , suggesting hemodynamic changes in both measures are interchangeable. Using a cut-off value of a change in CI of 15%, only PPV predicted fluid responsiveness . SVV weakly predicted fluid responsiveness in this cohort . While there was a trend toward reduction in \u0394V peak this was not different between those defined as fluid responders by CI (repeated-measures ANOVA P = 0.124) and \u0394V peak prior to fluid bolus did not predict a CI response .Twenty-six patients (mean age 40 (13), 15 male:11 female) were assessed, mean APACHE II 23 (4) and SOFA 15 (2). Changes in CI and SVI were closely correlated (Baseline PiCCO parameters predict fluid responsiveness but the respiratory variability in \u0394V peak did not predict a CI response to fluid bolus in this cohort. PPV may be a more suitable PiCCO index for assessing fluid requirements in patients with ALF than SVV."} +{"text": "Panels A and B in Supplementary Figure S1Analysis of host plant volatile composition with cactus rot stage. Cacti were either uninoculated (NI) or inoculated and fermented for one to nine weeks. Peak numbers correspond to the list of volatiles. (A\u2013D) Typical gas chromatograms of barrel, prickly pear, organ pipe and agria headspace (respectively) from uninoculated or representative fermented samples .(PDF)Click here for additional data file."} +{"text": "Increased ventricular and arterial stiffness is associated with diastolic dysfunction (DDFx) in patients with heart failure and preserved systolic function. Limited information is available regarding the impact of aortic biomechanics and the ventricular-vascular coupling (VVC) on DDFx in those patients with advanced ischemic cardiomyopathy (ICM). In addition, it is not known if cardiac magnetic resonance (CMR) measurements of LV remodeling (sphericity and scar burden) can also contribute to prediction of DDFx in these patients. We sought to examine the relationship between aortic biomechanical properties , ventricular-ventricular coupling , LV remodeling assessed by CMR and diastolic function assessed by echocardiography in patients with advanced ICM.Patients were selected if they had undergone TTE and CMR studies within 7 days (median=1 day). 354 patients with LVEF \u2264 40% and \u2265 70% stenosis in \u22651 coronary artery but without prior mitral valve surgery, fused E/A waves, atrial fibrillation or > moderate mitral regurgitation were screened. Of those, 84 patients were excluded due to poor CMR image quality from artifacts and/or suboptimal temporal resolution. A total 270 charts were reviewed for demographic and laboratorial data. Diastolic function assessment was performed as per guidelines. Aortic biomechanics were measured using previously validated software using semi-automated tracing of aortic contours with phase-contrast images and through-plane velocity encoding of the ascending and descending aorta. CMR evaluation also included long and short axis assessment of LV sphericity and function respectively on balanced steady state free precession images along with assessment of myocardial scar (on phase-sensitive inversion recovery DHE-CMR sequence ~ 10-20 minutes). Multivariate linear regression analysis was done to identify the independent predictors of DDFx.Males represented 76% of the cohort with a mean age of 62 \u00b1 10 years. Mean LVEF was 23 \u00b1 5% and DDFx was classified as either: stage 1 (44%), stage 2 (25%) or stage 3 (31%). The independent predictors of impaired diastolic function (stage > 1) are listed on Table In patients with advanced ICM, CMR assessment of VVC, LV sphericity and scar burden are independent predictors of DDFx. Aortic biomechanical properties are not independently associated with diastolic dysfunction.None."} +{"text": "The purpose of this in vitro study was to evaluate the sealing ability of the readymade temporary filling and hand mixed materials by assessing coronal microleakage.Standardized access cavities were prepared in 80 intact human permanent premolar teeth. They were divided randomly into four experimental groups (n=20). The teeth were restored using one of the temporary materials including Cavisol, Litrak, Zinc phosphate cement, Zinconol (IRM). Thermocycling was applied on the specimens. Methylene blue dye was applied and penetration was evaluated under stereomicroscope. Grading of the microleakage pattern was from 1 to 3, with 3 providing the best seal. Results were analyzed using one-way ANOVA test (P<0.05).Microleakage of Cavisol and Litrak samples achieved grade 3; whereas zinc phosphate cement and Zinconol samples absorbed the dye into the bulk of the materials. Cavisol was found to exhibit the best seal amongst the four tested materials followed by Litrak, zinc phosphate cement, and Zinconol. There was a statistically significant difference in the microleakage scores obtained between the materials (P<0.01).Among the four materials tested, readymade temporary filling provided the best sealing ability over hand-mixed. This study emphasizes the importance of correct placement and sufficient thickness of temporary filling materials in endodontic access cavities to ensure a tight seal. Loss of integrity of coronal tooth substances and invasion of microorganism into dentine and pulp space play an important role in pulpal and periradicular diseases. Inadequate coronal seal allow biological contamination and penetration of saliva, nutrients, chemicals and also microorganisms and their byproducts. Coronal microleakage appears to be of equal or greater clinical relevance as a factor in endodontic failure than apical leakage due to risk of recontamination 2]3]4][3][4]2][[4][3][4][3][4]2]. A recenA coronal filling material is considered effective when it is able to fulfill certain properties including an effective seal of tooth margins, lack of porosity and dimensional stability to thermal changes, good abrasion and compression resistance, ease of insertion and removal, compatibility with intra-canal medicament and good aesthetic appearance 8]9].[9].8][9].[9].8][9Several studies evaluating the microleakage of temporary restorative materials have been conducted and the used techniques were mostly assessed seal ability using dye penetration with either thermal cycling or load cycling procedures 9]10][1[10][19][[1[10]1912]13].[13].12][.[13].12]Most of the conducted studies focused on the sealing ability of Zinconol (IRM) which exhibited gross microleakage ; Zmener The aim of this study was to evaluate the sealing ability of readymade and hand mixed temporary filling materials by conducting microleakage tests using methylene blue dye.Eighty caries free extracted human maxillary and mandibular premolars that had been stored in 10% formalin were cleaned of soft tissue and debris, rinsed overnight in running water and then immersed in deionized water for 24 hours.Standardized coronal access cavities to the pulp chamber were prepared in the occlusal surfaces with the aid of a template measuring 4mm\u00d74mm; access was made using a high speed air turbine under water coolant with a round bur for initial entry and a diamond fissure bur to extend the preparation to the desire occlusal outline. All teeth were irrigated using 5% sodium hypochlorite to remove remaining smear layer, pulp tissues and other debris inside the pulp chamber. The prepared openings were air dried and cotton pellet were placed on the floor of the pulp chamber. A periodontal probe was used for measuring the final depth cavity and assuring that it could accommodate at least 4mm of the temporary filling material. The teeth were divided randomly into four groups of 20 teeth each . All matAll samples were screened visually and then under the stereomicroscope. Amongst four tested materials, Cavisol showed the least microleakage, followed by Litrak, zinc phosphate and Zinconol. The results showed there were statistically significant differences between tested materials. A Kruskal Wallis (ANOVA) test revealed a significant difference between four materials (P<0.01).Post hoc test using Mann-Whitney U tests revealed a significant difference between the readymade and hand mixed one (P<0.01).Cavisol and Litrak samples showed the least microleakage , followed by zinc phosphate cement which had leakage grade 1 (85%) and grade 2 (15%).Zinconol samples had the worse results with 100% of samples scoring grade 1 .MicrograIn this in vitro study, readymade filling materials (Cavisol and Litrak ) were compared to hand mixed (zinconol and zinc phosphate) temporary restorative materials. The experiment was conducted on extracted intact premolars with 4mm thickness of temporary restorative materials conforming to previous reports 18]19],[19],18],18]To prevent weakening of the tooth, coronal structure should be preserved whenever possible , whereasThe present study utilized thermal cycling procedure to simulate intraoral conditions. The temperature range of 55\u00b12\u00baC and 5\u00b12\u00baC used in this study corresponds to the extremes of temperatures that could be experienced in the oral environment 18]19].[19].18][.[19].18]Our observations revealed that all tested materials leaked to some extent. Different authors have reported conflicting results concerning the ability of readymade temporary filling materials such as Cavit and hand mixed materials to prevent coronal microleakage. Some revealed that Cavit a readymade material had the best sealing ability whereas IRM as hand mixed showed the maximum dye penetration . Other iReadymade temporary filling had superior sealing ability over hand-mixed. This emphasize the importance of material and correctly placing a sufficient thickness of temporary filling materials in endodontic access cavities to ensure a tight seal. However, further research such as a long-term study may provide better evidence."} +{"text": "To the Editor: African swine fever (ASF) is a highly contagious and deadly hemorrhagic disease of domestic pigs caused by African swine fever virus (ASFV), a double-strand DNA virus of the family Asfarviridae and genus Asfivirus gene was amplified by using p72U/p72D primers analysis of 2010\u20132012 ASFV p72 nucleotide sequences from Tanzania in GenBank showed 100% nucleotide identity with the Georgia 2007/1 ASFV isolate (GenBank accession no. FR682468) at nt positions 103,594\u2013104,070.All obtained ASFV p72 nucleotide sequences were 100% identical. BLAST (The Georgia 2007/1 ASFV isolate was detected in Georgia in 2007 and has caused ASF outbreaks in Armenia, Azerbaijan, and Russia ("} +{"text": "Dilated cardiomyopathy (DCM) is a common cause of heart failure (HF) and the most common diagnosis in patients referred for cardiac transplantation. Genetic studies in DCM have identified mutations in over 30 genes. To date, there is only scarce data regarding the interdependencies between the various genotypes and their impact on cardiac morphology and myocardial tissue appearance. Since non-contrast cardiac magnetic resonance (CMR) is considered the reference method for morpho-functional analysis and late gadolinium enhanced (LGE) CMR method of choice for myocardial tissue assessment, we sought to investigate the characteristics of LGE in asymptomatic and symptomatic carriers of known cardiomyopathy mutations and their clinical associations.Thirtyeight carriers of lamin A/C (LMNA), cardiac troponin T (TNNT2) or myosin binding protein C (MYBPC3) gene mutations were investigated with a 1.5T clinical scanner (Philips Achieva). Short axis slices covering entirely both ventricles were acquired using standard SSFP-sequences for measurement of LV and RV volumes and ejection fraction (EF). Focal myocardial fibrosis was assessed in LGE CMR images acquired 10 minutes after i.v. injection of 0.2 mmol/kg Gd-DTPA (Magnevist). Standard SSFP and LGE CMR images were assessed by different observers who were blinded to measurement results and clinical data. Associations between clinical characteristics, gene mutation and LGE were analyzed.LGE was present in 19 of 30 (63%) MYBPC3 carriers, all 6 (100%) TNNT2 carriers and 1 of 5 (20%) LMNA carriers. Midwall patterns of LGE located in the basal and/or mid-ventricular septal wall were the most commonly seen patterns in all gene mutations . The presence of LGE was positively associated with age . No further significant correlations were found between the presence, pattern or location of LGE and gender, cardiovascular risk factors, baseline systolic LV function or conduction abnormalities. Of all patients with known mutations, 24/38 (63%) were asymptomatic. No significant difference in LGE features was found between symptomatic and asymptomatic mutation carriers .LGE, predominantly presenting with basal and/or mid-ventricular septal midwall patterns, is a common finding in cardiomyopathy patients with known gene mutations. Its presence seems to be independent of the symptomatic stage of the disease or its phenotypic manifestation.none"} +{"text": "Nasal glucocorticosteroids (NGCS) are considered first-line therapy for allergic rhinitis and are effective therapy for nonallergic rhinitis, especially vasomotor rhinitis and nonallergic rhinitis with eosinophilia syndrome -4. NasalNGCS are helpful in reducing the amount of rebound congestion in patients with perennial allergic rhinitis with RM. They also decrease nasal mucosal edema, recruitment of neutrophils and mononuclear cells, cytokine production, and late-phase nasal mediators. They may offer a protective benefit from the development of RM. Oxymetazoline may also decrease turbinate enlargement, thereby permitting better nasal distribution of NGCS.Data are limited about efficacy and safety of OXY used with NGCS for nasal congestion (NC) not adequately responding to recommended doses of NGCS.Patients had at least a 1-year history of perennial allergic or nonallergic rhinitis documented by skin testing in the medical record before study enrollment. Those receiving allergen immunotherapy were on a stable maintenance regimen for at least 30 days before the first study visit and remained on the same dose during the study. Institutional review board approval and informed consent were obtained before initiation of the study.Patients completed twice daily diary cards pertaining to their nasal symptoms: 0 (not noticeable); 1 ; 2 ; 3 . All had a nasal congestion score of 1.5 or greater at the screening visit (day 7) and an average nasal congestion score of 1.5 or greater at the baseline visit (day 1). The average nasal congestion score was calculated from during the last 3 days before baseline and the morning of the baseline visit .\u03bcg every night or nightly and OXY 0.05% 2 sprays per nostril (s/n) 2 times daily versus MNS 100 \u03bcg every night or nightly plus saline placebo (PL) 2 s/n 2 times daily 100 Patients completed diaries for NC and quality of life (QOL) and underwent intranasal examination for nasal and turbinate changes and rhinorrhea. Patients continued on NGCS after study termination.P value comparing the 2 groups on days 18 to 20 was 0.12. Slightly more nasal erythema occurred with OXY. No RM or other side effects occurred in either group.Twenty-three patients met inclusion criteria. The MNS plus OXY cohort versus MNS plus PL achieved an averaged NC score of 1.68 versus 2.06 (NS), respectively is stratified into 5 categories: activities, practical problems, nose symptoms, eye symptoms, and other symptoms. Patients rate symptoms from 0 (not troubled) to 6 (extremely troubled).P = 0.04 (Figure Among the 5 Mini RQLQ indicators, the MNS plus OXY cohort reported the biggest benefit on activity measures, 4 Figure . The treMNS plus OXY combination for 20 days decreases NC and improves activity QOL indicators without significant side effects or risk for RM. Most Mini RQLQ data are not statistically significant. However, quality of life activity measures are statistically significant with sleep improvement most notable. Relief of nasal congestion trends toward improvement both in nasal congestion scores and QOL scores. It is unclear why most improvement is seen on days 5 to 7. A larger study powered with 145 patients is necessary to prove statistically the effectiveness and safety of these 2 medications used together. There are several papers presented at the American Academy of Allergy Asthma and Immunology, New Orleans, 2010, meeting which demonstrate effectiveness of MNS plus OXY versus one or the other without appreciable side effects -13. It i"} +{"text": "Interferon alpha (IFN\u03b1) has immune stimulatory actions implicated in its pulmonary toxicities. We describe deterioration of idiopathic pulmonary fibrosis (IPF) associated with IFN\u03b1 treatment for chronic hepatitis C in a 58\u2009year old woman culminating in a fatal suspected acute exacerbation of IPF (AE-IPF). Caution should be exercised in the use of IFN\u03b1 in subjects with concomitant IPF given its known immunostimulatory effects and possible role in this suspected AE-IPF. Pulmonary toxicity is a recognised complication of interferon alpha (IFN\u03b1), thought to be related to its immunostimulatory effects. IFN\u03b1 has been shown to influence the immune system along multiple pathways including augmentation of proinflammatory and profibrotic cytokines -related Child-Pugh A liver cirrhosis developed an oesophageal variceal bleed. Hepatitis C was associated with moderate inflammatory activity (Metavir score A3F4), previous hepatic decompensation with ascites and F3 oesophageal varices treated with prophylactic ligation. She had a history of tobacco-related chronic obstructive pulmonary disease (COPD). Eighteen months before admission, high-resolution computed tomography (HRCT) of the chest demonstrated lower zone predominant subpleural interstitial fibrosis with honeycombing consistent with UIP/IPF (see Table Pseudomonas aeruginosa and non-multiresistant methicillin-resistant Staphylococcus aureus (nmMRSA). On day 6, intravenous hydrocortisone was commenced. On Day 7, bronchoalveolar lavage was performed and specimens again grew nmMRSA, however tested negative for all other pathogens including Pneumocystis jiroveci (carinii). Transthoracic echocardiogram demonstrated normal left ventricular function. On day 10, there was further deterioration in oxygenation and she required high flow oxygen via mask to achieve oxygen saturations of 90%. Pulse intravenous methylprednisolone was commenced however her respiratory status continued to deteriorate. Escalation of therapy including endotracheal intubation was thought to be futile and she died on day 14.On admission, she received intravenous octreotide, prophylactic ceftriaxone and endoscopic variceal banding. On day 3, she developed progressive hypoxaemia, requiring high flow nasal prong oxygen to achieve oxyhaemoglobin saturations of 96%. She was febrile to 38.3\u00b0C. HRCT of the chest demonstrated diffuse ground-glass change superimposed on markedly deteriorated bilateral pulmonary fibrosis with honeycomb changes and radiologic (within 2\u2009months) evidence of deterioration. The 6% decline in FVC was felt to be within the limits of variability, although recent data have shown that FVC decline of >5% predicts a doubling of one year mortality and subsequent bronchoscopy (performed because of progressive hypoxia) did not reculture Pseudomonas. Furthermore, her clinical course was marked by relentlessly progressive, antibiotic-unresponsive, hypoxic respiratory failure without evidence of sepsis, features much more consistent with AE-IPF than nmMRSA pneumonia. While exclusion of infection forms part of Collard\u2019s proposed criteria for definition of AE-IPF (Collard et al. The acute respiratory decline during the described admission was suspected to be secondary to AE-IPF. Although Given that IFN\u03b1 is now part of standard therapy for chronic HCV infection and the rising incidence of UIP/IPF in Western countries (Navaratnam et al."} +{"text": "Transplant rejection affects the course and survival after heart transplantation (HTx). As a non-invasive alternative to myocardial biopsy, which is the gold standard used for screening and for the diagnosis of rejection, regional left ventricular (LV) myocardial motion analysis has been suggested. But myocardial biopsy is invasive and its diagnostic value is restricted by high sample errors due to the patchy distribution of early rejection. Alterations of regional wall motion , especia27 heart transplant recipients without signs or former episodes of rejections or relevant transplant vasculopathy (LVEF=63\u00b15%) were examined using phase contrast MRI (Tissue Phase Mapping) to assess 3D-myocardial velocities. Systolic and diastolic peak radial and long-axis velocities were evaluated in each LV segment. These functional parameters were analyzed with respect to time after HTx and CIT.Shorter times after HTx (n=8 patients <12 month after HTx versus n=19 >12 month) resulted in an increase of systolic radial velocities and a reduction of diastolic long-axis velocities . With longer CIT (>155 minutes) an increase of peak systolic radial velocities and a reduction of diastolic long-axis velocities were demonstrated compared to those with shorter CIT (<155 min) Program \"Advanced Cardiovascular MRI Research Center\", Deutsche Forschungsgemeinschaft (DFG): Grant # FO 507/2-1, FO 507/3-1"} +{"text": "Defective IFN signaling results in loss of innate immunity and sensitizes cells to enhanced cytolytic killing after Vesticular Stomatitis Virus (VSV) infection. Examination of the innate immunity status of normal human bronchial epithelial cells Beas2B and 7 lung cancer cells revealed that the abrogation of IFN signaling in cancer cells is associated with greater sensitivity to VSV infection. The disruption of the IFN pathway in lung cancer cell lines and primary tumor tissues is caused by epigenetic silencing of critical interferon responsive transcription factors IRF7 and/or IRF5. Although 5-aza-2\u2032-deoxycytidine treatment fails to reactivate IRF7 and IRF5 expression or protect cells from VSV infection, manipulating IFN signaling by altering IRF expression changes the viral susceptibility of these cells. Lung cancer cells can be partially protected from viral killing using IRF5+IRF7 overexpression, whereas IFN pathway disruption by transfection of siRNAs to IRF5+IRF7 increases cells' vulnerability to viral infection. Therefore, IRF5 and IRF7 are key transcription factors in IFN pathway that determine viral sensitivity of lung cancer cells; the epigenetically impaired IFN pathway in lung cancer tissues provides potential biomarkers for successful selective killing of cancer cells by oncolytic viral therapy. As the leading cause of cancer-related mortality in both men and women, lung cancer is responsible for well over 1 million deaths worldwide annually. Although diagnosis and treatment have been improved, the five-year survival rate is only 14% largely due to the failure of tumor debulking surgery and systemic chemotherapy. The improvement of lung cancer treatment is a major public health goal. Recently, naturally occurring or genetically engineered oncolytic viruses, including measles virus, Newcastle Disease Virus (NDV), VSV, adenoviruses, reovirus and Herpes simplex virus offer an effective and promising alternative therapeutic approach to fight this disease Type I IFN signaling pathway is activated by VSV infection as first line innate immune response to protect normal tissues from viral killing, and therefore tumor cells that have lost their antiviral reactivity represent selective targets for VSV. The primary response upon viral infection and uptake of double-stranded RNAs is TLR3 activation which is mediated by IRF-3, cJUN/ATF-2, and NF\u03baB, thereby inducing the production of immediate-early response genes primarily IFN\u03b2. Those early response IFNs bind to type I IFN receptors (IFNAR) in an autocrine or paracrine manner to activate STAT1 and induce expression of secondary antiviral response genes including the transcription factor IRF7 which then promotes the expression other IFN stimulated genes (ISGs). Finally, the tertiary transcriptional wave of IFN\u03b1 establishes an antiviral state The impairment of IFN signaling is linked to an enhanced risk of tumor development IRF7 expression corresponded to aberrant promoter hypermethylation of CpG islands within its promoter and was also identified as one of methylation-silenced genes in several cancer types including lung, hepatocellular, gastric and pancreatic cancers IRF5 was similarly observed in hepatocellular and gastric cancer The IFN-inducible transcription factors, IRFs, are essential mediators of the IFN-response. Lack of IRF7 and/or IRF5 in several lung cancer cell lines. Similarly, when we investigated DNA from fresh frozen lung cancer tissues, we observed promoter methylation of IRF7 and/or IRF5, suggesting that their IFN antiviral response was also epigenetically silenced as functional IRF7 and IRF5 are required for an intact IFN pathway. However 5-aza-dC treatment failed to reactivate IRF5 or IRF7 expression or rescue lung cancer cells from VSV infection. Altering innate immunity by manipulating IRF expression changed viral susceptibility of normal Beas2B bronchial epithelial cells or lung cancer cells. Cells without a functional IFN response are partially protected from virus following IRF5 and IRF7 overexpression, whereas disruption of IFN signaling by targeting IRF5 and IRF7 using siRNAs increased Beas2B cells' vulnerability to the cytolytic effects of VSV. Therefore, IRF5 and IRF7 are pivotal factors in IFN pathway that determine the viral sensitivity of the cells to oncolytic viruses. The highly selective VSV killing of lung cancer cells with an impaired IFN pathway due to epigenetically downregulation of IRFs indicates that these genes are ideal biomarkers for determining the susceptibility of tumors to oncolytic viral therapy.Although cancer cells, with their IFN-pathway-abrogated, may have acquired a growth/survival advantage over their normal counterparts, they simultaneously compromise their antiviral protective ability. Here, new therapeutic paradigms involving oncolytic RNA viruses to target the defective innate immune system in cancer cells are being explored. We found that the sensitivity to oncolytic VSV was strongly and significantly associated with the disruption of the IFN signaling pathway. A failure to up-regulate ISG expression upon dsRNA stimulation indicated a weakened antiviral response sensitizing lung cancer cell lines to VSV-induced oncolytic cell death. However, not all lung cancer cells can be killed by VSV infection, as some of them possess a relatively normal innate immunity pathway and therefore are resistant to VSV-mediated viral killing. Bisulfite sequencing revealed promoter hypermethylation of either ras transformed derivative cells and 2 tumorigenic clones revealed that as IFN signaling activity decreased the sensitivity to VSV killing increased during lung tumorigenesis . Because functional inactivation of IFN pathway has been a common trait of many cancers, we used 7 long-term lung cancer cell lines to study the changes in their innate immune system.The IFN pathway controls the cellular response to viral infection and dsRNA. Cells that have a fully functional innate antiviral system are able to protect themselves against viruses, largely due to the induction of IFN signaling cascade. We have shown that the IFN pathway was epigenetically inactivated in fibroblasts from LFS patients after spontaneous immortalization IFN\u03b1, IFN\u03b2, IRF7, IRF5 and STAT1 were detected in CRL5800, CRL5807, CRL5872 and CRL5869 cells. Surprisingly, Q-RT-PCR demonstrated polyI:C inducible expression of 10 out of 14 ISGs in HTB182 and CRL5928 cells at similar levels to Beas2B indicating a relatively normal antiviral response in those two lung cancer cells , whose expression was distinctively elevated >3-fold in response to poly I:C treatment only in VSV-resistant cell lines even without exogenous IFN\u03b1 pretreatment . In contll lines . Westernll lines . No signIRF7 promoter hypermethylation, which resulted in decreased antiviral defenses of the respiratory epithelium IRF7 could also be the cause of IFN pathway disruption in lung cancer cell lines. DNA sequencing of sodium bisulfite-treated genomic DNA revealed IRF7 promoter hypermethylation in 2 lung cancer cell lines (CRL5810 and CRL5869), which suggests that epigenetic silencing of IRF7 has played a role in the disruption of IFN signaling in these cell lines . Similar resistance to 5-aza-dC treatment was found for the other IRF7-methylated lung cancer cell line CRL5869 (data not shown). As a result, extended demethylation treatment with 5aza-dC was unable to affect the VSV sensitivity of CRL5810 cells and IRF7 (siIRF7) were applied to suppress IFN response in cells with active IFN pathway. Compared to Beas2B cells transfected with control scrambled siRNA, cells transfected with siIRF5 or siIRF7 resulted in a decrease in IRF5 and IRF7 induction after polyI:C treatment while transfection of both siIRF5+siIRF7 totally eliminated IRF activation . TransfeIn summary, the loss of IFN signaling is necessary and sufficient for increased sensitivity to killing by oncolytic viruses. The manipulation of the IFN pathway through the transcription factors IRF5 and IRF7 altered the cells' response to VSV infection. Analysis of the IFN pathway using these IRF methylation biomarkers may provide new theranostic biomarkers for determining a patient's sensitivity to oncolytic viruses.A functional innate immune antiviral system protects cells against viral infection, mainly due to the induction of the defensive IFN signaling cascade, which appears to be the basis for virus resistance and immune stimulatory properties. Here, we have demonstrated the strong and convincing inverse relationship between effective innate antiviral response and oncolytic viral susceptibility using lung cancer cell lines. Moreover, transcription factors IRF5 and IRF7 were identified as critical regulators of innate immune system and useful biomarkers for oncolytic virus susceptibility in lung cancer cells as both of them have to be inducible for a functional IFN pathway. Inactivation of either IRF5 or IRF7 weakened the antiviral response with the most significant loss when both IRFs were epigenetically silenced in CRL5810 cells. The varying extent of cytolytic death was related to the differential severity of IFN pathway defects. Despite the presence of exogenous IFN\u03b1, CRL5810 cells are non-responsive and remain vulnerable to VSV killing corresponding to the complete disruption of IFN pathway activity.IRF5 and IRF7 at CpG islands of promoter regions in lung cancer. Pharmacological targeting of aberrant epigenetic changes by demethylation agents has shown clinical efficacy in several hematologic malignancies Loss of tumor suppressor gene expression by aberrant promoter methylation is an early and common epigenetic event during the onset and progression of cancer Although basal levels of most ISGs were reduced in all the lung cancer cell lines compared to normal bronchial epithelial cells, oncolytic viral sensitivity is only associated with polyI:C-inducible expression levels of certain key ISGs such as IRF7 and IRF5. However, IRF5 and IRF7 overexpression is not sufficient to completely restore IFN pathway and only partially rescued cells from viral infection due to deficiency of other important ISGs. Failure to induce Ser727 phosphorylation upon polyI:C treatment suggested additional innate immunity defects in several VSV sensitive cell lines . In addiWe presented evidence that functional inactivation of IRF5 and IRF7 is the major mechanism to disrupt IFN signaling in lung cancer cells. Nevertheless, various malignancies harbor diverse molecular defects of this pathway. Deregulated JAK3 and RNase L pathways in LNCaP prostate cancer cells A failure to activate innate immunity response upon oncolytic RNA virus infection leads to the highly selective clearance of IFN-nonresponsive tumor cells. In addition to VSV, other RNA viruses such as NDV Although a reduced antiviral response may be a common feature of a broad range of cancers, the oncolytic efficacy of naturally occurring RNA viruses may still be relatively low, as some tumors manifest robust innate immunity responses that inhibit viral replication and spread. To overcome this obstacle, damping of cellular IFN responses in cancer cells by IFN-antagonist, such as influenza NS1 or inhibition of IFN-stimulating kinase (mTORC1) have been demonstrated to be effective strategies to augment therapeutic viral activity IRF5 and IRF7 as a theranostic strategy for oncolytic virus VSV treatment of lung tumors. Because not all of the lung cancer patients may benefit from VSV oncolytic viral therapy due to relatively normal function of the IFN pathway in some cancer cells, using IRF-promoter methylation as theranostic method for developing ISG-promoter related biomarkers is capable of assessing the susceptibility of each specific cancer case. Its absence limits the successful application of oncolytic therapy, causing delay of other more effective treatment and unnecessary side effects in VSV resistant patients The abrogated IFN response in more than 80% of human cancers favors the selective viral replication and cytotoxicity in those tumor cells and makes them ideal targets for oncolytic virus treatment Beas2B cell line was derived from immortalization of normal human bronchial epithelial cells (NHBE) with SV40-adenovirus E1a hybrid virus. Beas2B cells were used as normal control cells as they retain many properties of NHBE including potential for terminal differentiation; they are believed to represent the normal progenitor cells for lung carcinomas Twenty fresh frozen primary non-small-cell lung carcinoma (NSCLC) tissue samples and buffy coats from the same lung cancer patients were obtained from The Ontario Tumour Bank, Toronto, Ontario, Canada. This included 12 adenocarcinomas and 8 squamous cell carcinoma tissues. Detailed information of those samples is listed in 5-aza-dC treatment was done followed the protocol described earlier Total RNA was extracted from each experiment using the QIAGEN RNeasy Kit . Two \u00b5g total RNA was reverse transcribed into cDNA using Superscript II . Q-RT-PCR was performed using SYBR Green PCR Detection kit as described previously Western blots were performed as described Genomic DNA for lung cancer cell lines, primary lung cancer patient tissues, and buffy coat DNAs from those patients were prepared using QIAamp DNA kit . Genomic DNA (0.5 \u00b5g) was denatured and bisulfite converted using EZ DNA methylation-GOLD kit . The bisulfite modified genomic DNA was suspended in 10 \u00b5l of water and 2 \u00b5l of DNA was amplified by PCR with two nested PCR reactions. The annealing temperature was 56\u00b0C for the first PCR and 58\u00b0C for the second PCR. The two sets of primers for IRF7 are:5\u2032 GTAAGGGTTTTTGTCGTAGTAGACGTTAG;F1 5\u2032 AACGTAATAATTCATACCTATAATCCCAAC;R1 5\u2032 GGTTATAGGTGTGATTGTAGGTGTG;F2 5\u2032 CCCTAAACTATAATAAAATAACTCCATCTC.R2 The two sets of primers for IRF5 are:5\u2032 TGATTGGAAGGCGATTTAGG;F1 5\u2032 AAAATCCCAAACCGACCGAA;R1 5\u2032 AGTGGGGAAGTATTTTATTTTTTTT;F2 5\u2032 CCCCTAAACAACTACTACTAAACTCC.R2 The PCR products were subjected to double-strand DNA sequencing using primer F2.4 cells per well and cultured overnight in the presence or absence of IFN\u03b1 . The cells were then infected with a low dose or high dose (MOI\u200a=\u200a5.0) of VSV (Indiana strain) for 1 hr. Virus-induced cytopathicity was determined the next day by modified version of MTT assay as described previously Cells were seeded in 96 well plates at a density of 1\u22122\u00d710pCMV-IRF7, pCMV-IRF5 and control vector pcDNA3.1 were stably transfected into CRL5810 cells followed by 200 \u00b5g/ml G418 selection as described previously Table S1Information about collected lung cancer tissue samples.(DOC)Click here for additional data file.Table S2List of primer sets used in Q-RT-PCR.(DOC)Click here for additional data file."} +{"text": "Genetic analysis of factors affecting risk to develop excessive ethanol drinking has been extensively studied in humans and animal models for over 20 years. However, little progress has been made in determining molecular mechanisms underlying environmental or non-genetic events contributing to variation in ethanol drinking. Here, we identify persistent and substantial variation in ethanol drinking behavior within an inbred mouse strain and utilize this model to identify gene networks influencing such \u201cnon-genetic\u201d variation in ethanol intake. C57BL/6NCrl mice showed persistent inter-individual variation of ethanol intake in a two-bottle choice paradigm over a three-week period, ranging from less than 1 g/kg to over 14 g/kg ethanol in an 18 h interval. Differences in sweet or bitter taste susceptibility or litter effects did not appreciably correlate with ethanol intake variation. Whole genome microarray expression analysis in nucleus accumbens, prefrontal cortex and ventral midbrain region of individual animals identified gene expression patterns correlated with ethanol intake. Results included several gene networks previously implicated in ethanol behaviors, such as glutamate signaling, BDNF and genes involved in synaptic vesicle function. Additionally, genes functioning in epigenetic chromatin or DNA modifications such as acetylation and/or methylation also had expression patterns correlated with ethanol intake. In verification for the significance of the expression findings, we found that a histone deacetylase inhibitor, trichostatin A, caused an increase in 2-bottle ethanol intake. Our results thus implicate specific brain regional gene networks, including chromatin modification factors, as potentially important mechanisms underlying individual variation in ethanol intake. Over 121 million Americans drink alcohol, while less than 10% of the population drinks excessively Despite such progress on identifying genetic influences in alcoholism, little progress at the molecular level has revealed mechanisms that mediate environmental influences on ethanol behaviors or alcoholism. It is well documented that environmental influences such as stress or exposure to conditional stimuli can modify ethanol drinking or cause recidivism in abstinent alcoholics. Understanding the molecular mechanisms underlying such environmental influences on ethanol behaviors would augment the genetic progress mentioned above.C57BL/6 (B6) inbred mice have been widely used as a model for studying alcohol abuse related behaviors and the genetic basis of alcohol abuse since these mice voluntarily consume large volumes of unadulterated ethanol Here, we demonstrate a remarkable degree of individual variation in ethanol drinking behavior across individual mice from the C57BL/6NCrl inbred line. We have performed whole genome expression profiling in individual mice to finely dissect molecular factors underlying this individual variation in ethanol drinking behavior. We hypothesized that an as yet unidentified non-genetic factor has caused long-lasting brain signaling alterations that influence ethanol preference and intake in these mice. By characterizing gene networks differentially expressed between ethanol preferring and avoiding mice, we have identified putative epigenetic mechanisms such as alterations in chromatin acetylation that may regulate gene transcription and influence drinking patterns. We expect that these studies may contribute to the identification of novel targets for pharmacotherapy in alcoholism.All procedures were approved by Virginia Commonwealth University Institutional Animal Care and Use Committee under protocol numbers AM10332 and AM10139, and followed the NIH Guide for the Care and Use of Laboratory Animals .Male C57BL/6NCrl mice (age 42\u201349 days) from Charles River Laboratories were habituated to the vivarium (5 mice/cage) for 1 week followed by individual housing for 1 week prior to beginning drinking experiments. Cages and bedding were changed weekly. Mice were housed in a temperature and light controlled room with free access to standard chow and water.Experiment 1: Voluntary two-bottle choice drinking was performed as described previously Experiment 3: For studies on ethanol preference in littermates, two cohorts of male B6 littermates were purchased as littermates from Charles River. Breeding at Charles River facility used two females paired to one male. Offspring were weaned at day 21, shipped to our vivarium at day 35 and remained housed as littermates until the beginning of the study. 10 litters were represented with 3\u20135 males per litter . Mice were individually housed for 7 days then presented with 10%(w/v) ethanol for 18 h/day in a two-bottle choice paradigm for 14 days.Experiment 4: Taste preference for a bitter or sweet solution was measured using quinine or saccharin vs. water in a two-bottle choice paradigm. Mice (n\u200a=\u200a16) were tested for ethanol preference for 14 days as in experiment 1 but without deprivation periods, and then allowed to rest for 7 days with only water and food available. Half the mice were given two bottles containing either a 0.1 mM quinine solution or tap water and the others given a choice between 0.033% saccharin/water. Bottles were alternated every other day. Consumption of quinine/water or saccharin/water was measured daily for 3 days (18 h/day) after which the other tastant was offered for 3 days in a counterbalanced design.Experiment 5: 18 male C57BL/6NCrl mice, acclimated to a reverse-light cycle, were used to test effects of trichostatin A on ethanol drinking. Voluntary ethanol drinking was initiated as before except these mice had access 24 hours/day, changed at 1200 h. Following 7 days baseline ethanol access, mice were divided into two groups balanced for baseline intake. Mice in the TSA group (n\u200a=\u200a9) were injected with 2 mg/kg TSA i.p. for 5 days. Control mice (n\u200a=\u200a9) were injected with vehicle. Mice had continuous ethanol access during the entire study .Experiment 6: Western blotting for acetyl-Histone-H4 was used to verify CNS activity of i.p. TSA. C57BL/6NCrl mice (n\u200a=\u200a12) received a single i.p. injection of 2 mg/kg TSA or vehicle. After 24 hours, nucleus accumbens was dissected for Western blotting as described below.The effect of TSA on ethanol metabolism was assayed in a separate group of mice (Experiment 7). Male C57BL/6NCrl mice received 2 mg/kg TSA or vehicle for 5 consecutive days. On day 6, all mice were injected with 2 mg/kg ethanol. Trunk blood was harvested from mice at 10, 60, 120 and 180 minutes following ethanol administration. Whole blood samples (20 \u00b5l) were placed into 20-ml headspace vials with 960 \u00b5l water and 20 \u00b5l 1-propanol internal standard. Samples were tested for ethanol concentration using a Hewlett Packard 5890A gas chromatograph equipped with a flame ionization detector, 2 meter 5% Carbowax 20M 80/120 mesh packed column and CTC Combi-Pal headspace autosampler. Data were acquired by Clarity software and analyzed by linear regression with no weighting. A 7-point calibration curve preceded the analysis and quality control ethanol standards were interspersed with each set of samples. Up to 3 replicates were analyzed from each animal and averaged if sufficient blood was collected.\u00b5g) was transcribed into double-stranded cDNA using the One-cycle Targeting and Control Reagent kit . Biotin-labeled cRNA was synthesized from cDNA, purified and fragmented according to manufacturer's instructions.Total RNA was extracted from PFC, NAc and VMB from individual mice in STAT 60 reagent according to the manufacturer's protocol. RNA concentration and quality was assessed by Experion automated electrophoresis . Total RNA was hybridized to a single microarray for each brain region . Samples were analyzed on oligonucleotide arrays (Mouse Genome 430A 2.0 array) containing >22,000 well-characterized genes and expressed sequence tags. Array hybridization and scanning were performed exactly according to manufacturer's protocols (Affymetrix).Microarray data were processed using GeneChip Operating Software v4.1 and normalized to a mean total hybridization intensity of 190. All raw microarray data is deposited at the Gene Expression Omnibus repository (GEO) under accession number GSE26506. All microarray data is MIAME compliant. Array quality was assessed by accepting arrays with scaling factor <3, 3\u2032\u20135\u2032-actin ratio <2, and by examining linearity and inter-chip correlations of intensity values. Arrays determined to be acceptable (57/58) were further analyzed using the Robust Multichip Average (RMA) low-level analysis algorithm to summarize probeset expression data www.ingenuity.com) and Bibliosphere (http://www.genomatix.de). These tools utilize biomedical literature associations to annotate genes with biological functions and cellular components. IPA also generates networks of interrelated genes based on their curated knowledge base.The Expression Analysis Systematic Explorer (EASE version 1.21) As an alternative to correlating gene expression to the average of the last 8 days of ethanol drinking data, we conducted a principal components analysis to reduce the number of covariates and identify the few principal components that account for a sufficient proportion of the variability and contain almost as much information as the full data set. The first 2 principal components accounted for 0.77 of the total variance. For each brain region, probeset-specific linear models predicting expression as a function of the two independent principal components were fit. An overall F-test was used for calculating P-values for each probe set level linear model. Genes significant at p<0.05 level were selected for further bioinformatics analysis and comparison with results from using average ethanol intake over the last eight days.d|\u22650.5 and q<0.05 Genes significantly correlated to ethanol drinking patterns in the present study, using a false discovery rate of 1%, were analyzed for overlap with previously published gene sets having expression significantly different between alcohol-preferring or non-preferring mouse models based on the criteria |Twenty-one C57BL/6NCrl mice (Exp. 2) voluntarily consumed ethanol as described above and brains were harvested six days following the last drinking session. Nucleus accumbens was homogenized in NP40 buffer with protease inhibitors . Western blotting was performed as described C57BL/6 (B6) male mice from Charles River Laboratories consumed a substantial amount of ethanol, 6.47\u00b10.99 g/kg/18 h, in the voluntary two-bottle choice (10% w/v ethanol or water) self-administration paradigm. Interestingly, mice showed a large degree of inter-individual variation in ethanol drinking , rangingTo assess whether taste discrimination was contributing to variation in ethanol drinking, we performed an additional experiment (Exp. 4) where mice (n\u200a=\u200a16) were assessed for ethanol drinking, followed by studies on preference for quinine (0.1 mM) or saccharin (0.033%). While there was some individual variation in quinine consumption, preference for saccharin or quinine showed no significant correlation to ethanol preference (data not shown). These results argue against sweet or bitter taste as a major contributing factor for the observed individual differences in ethanol preference.Additionally, we determined whether the observed individual variation in ethanol drinking was due to robust differences between litters (Exp. 3). In two separate cohorts consisting of 10 litters , ethanol intake over 14 days of baseline drinking did not differ between litters \u200a=\u200a1.258 p\u200a=\u200a0.2967, We hypothesized that persistent individual variation in ethanol drinking behaviors within an inbred strain might be caused by differential basal gene expression patterns generated by unknown environmental influences. Further, such differential gene expression patterns could be used to identify molecular pathways contributing to individual variation in ethanol drinking. We profiled 3 brain regions in individual mice: nucleus accumbens (NAc), prefrontal cortex (PFC) and ventral midbrain region (VMB). These brain regions were chosen because they are major components of the mesocorticolimbic dopamine reward pathway activated by ethanol and other drugs of abuse To identify molecular factors related to ethanol drinking behaviors, gene expression patterns were first correlated to a drinking template created from the last 8 days of ethanol access following a third round of ethanol deprivation see . This deTo identify gene expression correlated with drinking behavior, we also performed a principal component analysis on the daily drinking activity data to reduce the number of covariates, rather than averaging the drinking data over an interval. The first two principal components (PC) accounted for 77% of the total variance. For each brain region, probe set-specific linear models predicting expression as a function of the two independent principal components were fit. The number of transcripts which fit the linear model at a level p<0.05 was 547 in NAc, 670 in PFC and 725 in VMB . When thGene lists from microarray analyses were analyzed for over-representation of biological functions or gene network relationships using several different tools as described in The 889 transcripts from NAc correlating with average drinking values were analyzed by EASE Hdac11 (NM_144919.2), Rbbp4 (NM_009030.3), Rbbp7 (NM_009031.3), Sap18 (NM_009119.3) and Suds3 (NM_178622.4). All genes in this network were significantly correlated to average ethanol drinking are involved in DNA methylation events and work concurrently to repress transcription Biological functions of our gene lists were further investigated using the curated knowledge base in Ingenuity Pathway Analysis. This tool generates networks of genes with known interactions or biological function. One of the top networks generated through this process is shown in drinking . Rbbp4 aMyst3 (NM_001081149), Hdac11 and Ehmt2 (NM_147151) were significantly different in high versus low drinkers by t-test at p<0.05. Myst3, Myst histone acetyl transferase 3, is a member of a mouse histone acetyl transferase (HAT) complex that increases DNA transcription Rbbp4, Rbbp7), had intrinsic HDAC activity (Hdac11) or were involved in methylation of DNA (Men1 (NM_001168488), Mbd2 (NM_010773.2), Mll1 (NM_001081049), Ehmt2). These genes are believed to cause transcriptional silencing by removing acetyl groups from chromatin or increasing DNA methylation.The relative expression of select genes in this network in the top quartile of high ethanol drinkers (>7 g/kg/18 h) and the bottom quartile of low ethanol drinkers (<2 g/kg/18 h) are summarized in Ap2a1 (NM_025606), Ap2a2 (NM_007459), Ap2m1 (NM_207255), Dnm1 (NM_144516), Dnm1l (NM_028661), Vamp3 (NM_009498), and Vamp4 (NM_016796.3)) and synaptic vesicle biogenesis (Sh3gl2 (NM_019535), Sh3glb1 (NM_175141)) were generally positively correlated to ethanol intake. This suggests that synaptic vesicle recycling may be increased in mice prone to drinking greater amounts of ethanol. Conversely, low drinking mice had higher Bdnf (NM_001037955) expression. Bdnf may play a role to increase synaptogenesis in these studies as it has been implicated in plasticity from multiple drugs of abuse Further network analysis identified genes involved in synaptic vesicle formation and recycling see . Genes i+K+ ATPase activity and protein kinase activity , Grin3b (NM_028388)) and the kainite receptor (Grik1 (NM_010348)), as well as genes that bind (Htt (NM_010414)) or are regulated by glutamate receptors (Dlg4 (NM_001109752) aka Psd95). The NR2b subunit of the NMDA receptor was positively correlated to ethanol drinking with the lowest drinking mice having lower expression (Grik1) were correlated negatively to ethanol drinking. Tyrosine hydroxylase, the rate-limiting enzyme in catecholamine synthesis involved in the conversion of tyrosine to dopamine, was also positively correlated to ethanol drinking was chosen for its documented role in synaptic vesicle trafficking. In a separate cohort of mice (Exp. 2), RAB3A expression was determined in high and low drinking mice . RAB3A ebasal brain gene expression across multiple mouse strains with divergent ethanol drinking phenotypes. A large meta-analysis of this data identified gene expression correlated with ethanol drinking behavior across these multiple genetic models \u221234, Chi-square analysis) were also identified in the meta-analysis (see Stxbp1 (NM_009295)) has been previously identified as a putative candidate for an ethanol-drinking locus on Chromosome 2 \u221218, Chi-square) were also identified by the meta-analysis were in common with the meta-analysis results did not allow a definitive conclusion as to whether TSA affected the overall distribution of drinking values, since most animals showed increased drinking following TSA (not shown).et al. reported that within-strain preference variation was not correlated with gender or ethanol metabolism, and could not be altered by simple environmental disturbances Our studies here showed that C57BL/6NCrl mice express a striking degree of stable, inter-individual variation in ethanol drinking behavior with greater than 10-fold differences within a drinking session. We suspect these differences were generated by subtle environmental differences such as rearing behaviors The studies here employed a unique experimental design that allowed long-term measures of ethanol drinking behavior, ensured that such behavior was stable upon reinstatement, and permitted assaying gene expression differences in individual animals off ethanol. This allowed identification of expression patterns presumably \u201cpredictive\u201d of drinking behavior rather than simply resulting from such. However, even with the current design, we cannot totally eliminate the possibility that some of our gene expression results reflect, rather than cause, individual variation in drinking behavior. Additionally, even though the behavioral data showed extinction of an ethanol deprivation effect after several cycles of withdrawal and reinstatement , we cannThe current studies showed a potential role for epigenetic regulation of ethanol preference in B6 mice. In the NAc, genes with chromatin remodeling Gene Ontology function or classified in the HDAC complex had differential expression between high and low ethanol-drinking animals see . IntriguBdnf promoters, leading to decreased Bdnf transcription Inhibiting HDAC activity with TSA injections increased ethanol intake above baseline and vehicle-treated levels, supporting a role for chromatin modifications in the modulation of ethanol preference. The complex changes in chromatin modification gene expression made it difficult to predict how directly altering histone acetylation might affect drinking behavior. We suggest, however, that any TSA-induced change in ethanol drinking across individuals or as a population is supportive of our hypothesis regarding a role of chromatin modification in driving individual variation in ethanol intake. This data is the first to show modulation of drinking behavior by altering chromatin acetylation. However, evidence of ethanol-induced chromatin remodeling has been reported in hepatocytes Stxbp1 was previously identified as a candidate gene for a mouse Chr2 ethanol preference locus C. elegans and mice Our bioinformatics analysis of gene expression correlating with ethanol drinking also identified gene networks involved in synaptic vesicle biogenesis and recycling . Many ofBdnf mRNA levels and individual ethanol consumption. BDNF regulates multiple synaptic vesicle-related proteins, including several listed in Bdnf under-expression in Bdnf+/- mice caused increased ethanol consumption, consistent with Bdnf mRNA expression observed in the current study, where Bdnf is lowest in mice with the highest ethanol intake. We do not believe, therefore, that Bdnf expression levels seen in our studies were secondary to ethanol exposure itself. In support of this, we and other investigators have shown that acute ethanol injection (2 g/kg i.p.) in B6 or D2 mice increases Bdnf expression and that after 4 weeks of 2-bottle choice ethanol drinking, Bdnf is increased in the dorsal striatum versus non-ethanol controls Bdnf expression in the low drinking mice was possibly a causal factor in individual drinking behavior variance, rather than secondary to drinking behavior itself. We cannot currently exclude the possibility that ethanol drinking followed by withdrawal (4 days) caused the correlated changes in Bdnf expression. Together, these findings on Bdnf and synaptic vesicle-related gene expression are strong evidence supporting an important link between regulation of synaptic vesicles and individual variation in ethanol intake.We also identified an inverse correlation between Bdnf expression only being regulated in nucleus accumbens In the present study, many of the most robust gene expression changes were found in the nucleus accumbens and prefrontal cortex. Brain regional differential gene expression is not surprising considering the proposed different roles of each region in ethanol responses Importantly, we found significant overlap between our gene lists and a previously published meta-analysis of basal brain gene expression across mouse strains with differing ethanol preference In conclusion, the current experiments have described persistent inter-individual variation of ethanol drinking behaviors in B6 mice and, more importantly, they define gene expression networks that may underlie these individual differences. This study utilizes variation within an inbred strain to minimize genetic influences, isolating changes in gene expression due specifically to environmental factors. These experiments have identified several gene networks previously implicated in responses to ethanol in the NAc and PFC: glutamate signaling, BDNF and genes involved in synaptic vesicle function. Perhaps most importantly, our expression studies and behavioral analysis following histone deacetylase inhibition implicate epigenetic factors involving chromatin acetylation and/or methylation as contributing to environmental modulation of ethanol intake. Defining specific gene networks targeted by these epigenetic modifications is an important goal of ongoing studies. The novel findings presented here could contribute to understanding mechanisms involved in individual risk for alcohol abuse and alcoholism in humans. Future work will focus on characterizing the genesis and implications of gene network alterations and epigenetic modifications associated with variation in ethanol drinking.Figure S1Average Ethanol Intake Over 16 Days Of Access. Ethanol intake was significantly increased following repeated ethanol deprivations . Ethanol consumption did not differ from baseline after the third deprivation .(TIF)Click here for additional data file.Figure S2Genes Differentially Regulated In Ethanol Drinking Mice.A. Venn diagram overlapping and non-overlapping genes in each brain region significantly correlated to drinking at FDR<0.01. Region-specific expression patterns are represented as shaded circles (nucleus accumbens (NAc), dark; prefrontal cortex (PFC), open; ventral midbrain (VMB), light). B\u2013D. Hierarchical clustering of transcripts significantly correlated to ethanol drinking in the NAc (B), PFC (C) and VMB (D). Genes that overlap with the meta-analysis are labeled in blue. Genes that overlap with the principal component analysis are labeled in orange. Red color indicates higher relative expression and green indicates lower expression. Columns are arranged according to drinking behavior averaged over the last 8 days of intake, with low drinking mice on the left, progressing to higher drinking mice on the right.(TIF)Click here for additional data file.Figure S3Western Blot Analysis Of TSA Effects On Histone H4 Acetylation. Western blotting for acetyl-Histone-H4 was used to verify CNS activity of i.p. TSA. C57BL/6NCrl mice (n\u200a=\u200a12) received a single i.p. injection of 2 mg/kg TSA or vehicle. After 24 hours, nucleus accumbens was dissected for Western blotting for acetyl-histone H4 (upper panel) or GAPDH as a loading control (lower panel). Results verify increased H4 acetylation in NAc after TSA treatment.(TIF)Click here for additional data file.Table S1Average Ethanol Intake and Preference for six C57BL/6NCrl litters. Average ethanol intake and preference for six separate litters of male mice (n\u200a=\u200a4\u22125/litter) was calculated over 14 days of 24 h access to 2-bottle choice ethanol. There was no significant difference in intake or preference as reported in (XLSX)Click here for additional data file.Table S2Results from Correlation with Average Drinking. Data are gene lists and statistics for gene expression correlating with individual average ethanol intake from the last 8 days of ethanol access following a third round of ethanol deprivation. Data are presented for each brain region separately.(XLSX)Click here for additional data file.Table S3Gene Lists from Principal Component Analysis. Data are gene lists and statistics for gene expression correlating with ethanol intake data subjected to principal component analysis as in (XLSX)Click here for additional data file.Table S4Over-Represented Gene Categories From Gene Expression Significantly Correlated To Average Ethanol Drinking. Gene data from (XLSX)Click here for additional data file.Table S5Gene Categories Significantly Over-Represented From Principal Component Analysis In Ethanol Drinking C57BL/6NCrl Mice. Gene set data from (XLSX)Click here for additional data file."} +{"text": "NRG1) is linked to an increased risk of developing schizophrenia and cannabis dependence. Mice that are hypomorphic for Nrg1 (Nrg1 HET mice) display schizophrenia-relevant behavioral phenotypes and aberrant expression of serotonin and glutamate receptors. Nrg1 HET mice also display idiosyncratic responses to the main psychoactive constituent of cannabis, \u03949-tetrahydrocannabinol (THC). To gain traction on the molecular pathways disrupted by Nrg1 hypomorphism and Nrg1-cannabinoid interactions we conducted a proteomic study. Adolescent wildtype (WT) and Nrg1 HET mice were exposed to repeated injections of vehicle or THC and their hippocampi were submitted to 2D gel proteomics. Comparison of WT and Nrg1 HET mice identified proteins linked to molecular changes in schizophrenia that have not been previously associated with Nrg1. These proteins are involved in vesicular release of neurotransmitters such as SNARE proteins; enzymes impacting serotonergic neurotransmission, and proteins affecting growth factor expression. Nrg1 HET mice treated with THC expressed a distinct protein expression signature compared to WT mice. Replicating prior findings, THC caused proteomic changes in WT mice suggestive of greater oxidative stress and neurodegeneration. We have previously observed that THC selectively increased hippocampal NMDA receptor binding of adolescent Nrg1 HET mice. Here we observed outcomes consistent with heightened NMDA-mediated glutamatergic neurotransmission. This included differential expression of proteins involved in NMDA receptor trafficking to the synaptic membrane; lipid raft stabilization of synaptic NMDA receptors; and homeostatic responses to dampen excitotoxicity. These findings uncover novel proteins altered in response to Nrg1 hypomorphism and Nrg1-cannabinoid interactions that improves our molecular understanding of Nrg1 signaling and Nrg1-mediated genetic vulnerability to the neurobehavioral effects of cannabinoids.Neuregulin 1 ( Nrg1) is a neurotrophic factor that mediates its effects by binding ErbB receptor tyrosine kinases. Nrg1 regulates axonal guidance, myelination, and GABAergic and glutamatergic neurotransmission mouse which exhibits locomotor hyperactivity and protocol-dependent PPI deficits controls at 3\u20134 months of age, an effect that disappears by 5\u20136 months . Acute cannabinoid exposure promoted PPI facilitation in Nrg1 HET mice but PPI deficits in WT mice were at an age of post-natal day (PND) 31 \u00b1 2. The study was restricted to male mice as male Nrg1 HET mice appear more vulnerable to the effects of cannabinoids was suspended in a 1:1:18 mixture of ethanol:Tween 80:0.9% saline and injected intraperitoneally at a volume of 10 ml/kg. Mice were injected daily with either 10 mg/kg of THC or vehicle for 21 days. During this time mice were repeatedly behaviorally tested, the results of which are published elsewhere were euthanized by cervical dislocation, with both hippocampi dissected out and snap frozen on dry ice for proteomic analysis. Research and animal care procedures were approved by the University of New South Wales Animal Care and Ethics Committee and were in accordance with the Australian Code of Practice for the Care and Use of Animals for Scientific Purposes.At the commencement of the study adolescent male Protein extraction was performed using a protocol optimized for cytosolic proteins . Immobilized pH gradient strips were rehydrated with samples containing 600 \u03bcg protein, and samples were separated by isoelectric point (pI). Strips were equilibrated in SDS equilibration buffer and loaded onto SDS-PAGE gradient gels and separated by molecular weight using an ElectrophoretIQ3 system . Gels were fixed with methanol [25% (v/v)] and acetic acid [10% (v/v)] and visualized using Flamingo Fluorescent gel stain .p < 0.05) of spot volume were performed to determine the effect of genotype in vehicle-treated animals, the effect of THC administration in WT mice and the effect of THC administration in Nrg1 HET mice.Gels were analyzed using Phoretix 2D Expression software . Averaged gels were created for each experimental group and averaging parameters were set at 70%. Single factor ANOVAs before being eluted in 3 \u03bcL of matrix solution. Spots were analyzed using an Applied Biosystems QSTAR MALDI-TOF mass spectrometer . MALDI spectra were matched against the Swiss-Prot database using the MASCOT search engine with matches determined by molecular weight search score (MWS) and sequence coverage in conjunction with pI and molecular weight as estimated from gels.Protein spots that were identified as significantly altered were digested in 12.5 ng/mL trypsin and 25 nM NHTen ug of protein per lane was separated by electrophoresis using 10% precast NuPage gels and run at 110 V for 2 h. The samples were transferred to PVDF membranes. Membranes were sequentially incubated with Syntaxin-1A antibody then swine anti-rabbit secondary antibody and rabbit Peroxidase-Anti-Peroxidase and DAB . Membranes were stripped using Re-blot plus strong antibody stripping solution and incubated sequentially with Abcam rat monoclonal YL1/2 \u03b1-tubulin (TUBA) antibody then anti-rat IgG (H\u00feL) horseradish peroxidase conjugate and visualized using a Syngene G:Box.Nrg1 HET mice and WT control mice treated with or without THC. The averaged gels for WT vehicle, WT THC, Nrg1 HET vehicle and Nrg1 HET THC contained 870, 821, 761, and 742 spots respectively. 26 spots were significantly different between WT vehicle and Nrg1 HET vehicle mice. Of these spots, 17 proteins were identified using MALDI-TOF MS and the fold changes from control WT mice are listed in Table Nrg1 HET mice displayed increased expression of STX1A compared to WT mice respectively. Figure Nrg1 HET mice treated with THC displayed a distinct protein expression profile to WT mice exposed to the drug. Figure Nrg1 HET mouse administered repeated THC. Normalized spot volumes of three representative proteins are depicted in Figure THC induced changes in 28 spots and 23 spots in WT and Nrg1 HET mice, displayed altered expression of a number of proteins involved in the vesicular release of neurotransmitters including STX1A, syntaxin 7 (STX7) and \u03b2-SNAP; serotonergic neurotransmission including tryptophan 5-hydroxylase 1 (TPH1) and serotonin N-acetyltransferase (AA-NAT); growth factor expression and regulation including secreted protein acidic and rich in cysteine (SPARC) and GPC6, and; cell survival and regulators of inflammatory cytokines including cell death regulator Aven (AVEN), TNFAIP3-interacting protein 2 (ABIN2) and regulator of G-protein signaling 10 (RGS10). We replicated prior findings in rodents without genetic modification showing THC reduced the hippocampal expression of GSTM2 and affected the expression of heat shock proteins (here HSPA4). We also identified novel proteins changed in response to repeated THC exposure, that is, CALB2 and ARL1. Unlike WT mice, Nrg1 HET mice administered THC displayed altered expression of proteins involved in NMDA receptor trafficking to the synaptic membrane including GPSM2; lipid raft stabilization of receptors at the synaptic membrane including flotillin-1 (FLOT1); homeostatic responses to dampen excessive glutamatergic transmission, including NAPEPLD, and excitotoxicity and apoptosis including programmed cell death protein 2 (PCD2). Figure Nrg1 hypomorphism, THC and Nrg1-THC interactions.here HSPA. We alsoNRG1, a schizophrenia susceptibility gene, regulates neurotransmitter receptor expression and synaptic plasticity complex regulates exocytotic release of neurotransmitters from presynaptic terminals and alterations in SNARE mRNA and protein is observed in post-mortem schizophrenia brain and Golgi apparatus. These included \u03b2-SNAP, dynactin subunit 2 (DCTN2) and ADP-ribosyl cyclase 2 (BST-1). Transport of protein from the ER to the Golgi is important to protein sorting and the dispatch of protein to cellular locations. An increased expression of DCTN2 was also observed in Nrg1 HET mice. DCTN2 is a functional subunit of dynactin, a component of the dynein-dynactin system. DCTN2 overexpression inhibits dynactin, and therefore dynein functions such as dynein-dependent maintenance of membrane organelle distribution and regulators of growth factor protein expression, including SPARC and GPC6. FGF14 knockout mice, like Nrg1 HET mice, display locomotor hyperactivity, spatial learning deficits and impaired hippocampal long-term potentiation associated with lowered presynaptic vesicle docking (Wozniak et al., Nrg1 HET mice. The reduced level of FGF14 might be related to the increased expression of GPC6 and SPARC. Glypicans are heparan sulphate proteoglycans that act as co-receptors for growth factors and modulate fibroblast growth factor signaling (Paine-Saunders et al., Nrg1 and other interrelated growth factor-related proteins FG14, GPC6, and SPARC, worthy of further examination in future studies.Given that Nrg1 HET mice displayed altered expression of various proteins that influence cell survival and neuroinflammation including AVEN, ABIN2, and RGS10. Nrg1 exerts neuroprotective effects via inhibiting apoptosis triggered by various challenges (Chen et al., Nrg1 HET mice might confer greater vulnerability to apoptosis in these mice. Adolescent Nrg1 HET mice displayed a trend toward increased expression of the pro-apoptotic and inflammatory cytokine TNF-\u03b1 in the hippocampus and schizophrenia patients with a missense mutation in the transmembrane domain of NRG1 show heightened expression of TNF-\u03b1 from B cells (Marballi et al., Nrg1 HET mice, modulate the action of the TNF-\u03b1. ABIN-2 prevents TNF-\u03b1 mediated pro-apoptotic effects and its decreased expression in Nrg1 HET mice might reflect again a greater propensity to hippocampal apoptosis (Verstrepen et al., Nrg1 HET mice showed increased expression of RGS10, a protein which renders cells resistant to TNF-\u03b1 induced apoptosis (Lee et al., Nrg1 HET mice.The hippocampus of adolescent Nrg1 heterozygotes display an altered neurobehavioral response to cannabinoids (Boucher et al., Nrg1 mutant mice were more sensitive to the behavioral actions of acute THC compared to WT littermates in a sex-specific manner, with males being selectively affected but not females (Boucher et al., Nrg1 HET than WT mice (Boucher et al., Nrg1 HET mice maintained a persistent anxiogenic response to repeated cannabinoid exposure. Acute and repeated cannabinoid exposure selectively activated expression of Fos transcription factors in the lateral septum of Nrg1 HET mice but not WT mice (Boucher et al., Nrg1 hypomorphism confers vulnerability to the neurobehavioral actions of acute or repeated THC exposure in adolescence (Long et al., Nrg1 HET mice expressed after withdrawal of the drug. Further, repeated THC administration also promoted differential effects on CB1 receptor, 5-HT2A and NMDA receptor binding. Notably adolescent THC exposure selectively increased NMDA receptor expression in the hippocampus of Nrg1 HET but not WT mice. Given these findings it is perhaps not surprising that the impact of repeated THC treatment as measured by proteomics was quite distinct in Nrg1 HET mice vs. WT mice, with no overlap in differentially expressed proteins Table Our prior research shows that Nevertheless, our findings show some degree of overlap with previous examinations of THC effects on the rodent brain proteome (Quinn et al., 1 receptors are expressed on calretinin-positive GABA interneurons in the hippocampus (Marsicano and Lutz, Adolescent THC exposure decreased hippocampal levels of the calcium-binding protein CALB2. CBNrg1 HET mice treated with THC include those that affect synapse formation and the dynamics of dendritic spines. Nrg1 is a neurotrophic factor involved in spinogenesis through its modulation of NMDA receptor function (Li et al., Nrg1 hypomorphism might abnormally increase dendritic spine density in the hippocampus in response to THC as adolescent Nrg1 HET mice treated with THC displayed increased NMDA receptor binding in the hippocampus (Long et al., Proteins selectively altered in Nrg1 HET mice may reflect GPSM2 being incorporated into the macromolecular complex, lowering the observed expression of free, unconjugated GPSM2. Further, Nrg1 HET mice treated with THC showed a selective increase in FLOT1 expression in the hippocampus, a protein that helps stabilize lipid rafts in the membrane. FLO T1 mediates neurite branching and dendritic spine dynamics in the hippocampus (Swanwick et al., Nrg1 HET mice (Hirsch-Reinshagen et al., GPSM2 traffics intracellular NMDA receptors to the synaptic membrane and facilitates spinogenesis by forming a macromolecular complex with NMDA receptors and synapse associated protein 102 (Sans et al., Nrg1 HET mice might also increase the expression of the apoptotic marker PCD2 and anandamide synthesizing enzyme NAPEPLD (Howlett et al., Nrg1-cannabinoid interactions dysregulating the septohippocampal system. Increased excitation in the hippocampus of THC-treated Nrg1 HET mice might then influence downstream activity of the lateral septum, a region we have repeatedly shown to be selectively activated in Nrg1 HET mice in response to THC (Boucher et al., The increased excitatory transmission mediated by increased NMDA receptors in THC-treated Nrg1 signaling. Our findings also illuminate a potential constellation of molecular changes that may subserve the behavioral abnormalities that are observed in the Nrg1 transmembrane domain heterozygous mouse as well as their idiosyncratic response to repeated cannabinoid treatment. This may have implications for our overall understanding of genetic vulnerability to schizophrenia and to the exacerbation of psychosis sometimes caused by cannabis.Using a proteomic approach we have uncovered numerous novel proteins that may be subject to regulation by disturbed The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "GROWTH-REGULATING FACTOR (GRF) family of transcription factors, which are known to control cell proliferation in Arabidopsis leaves. In this work, we characterized the regulation of an additional target for miR396, the transcription factor bHLH74, that is necessary for Arabidopsis normal development. bHLH74 homologs with a miR396 target site could only be detected in the sister families Brassicaceae and Cleomaceae. Still, bHLH74 repression by miR396 is required for margin and vein pattern formation of Arabidopsis leaves. MiR396 contributes to the spatio-temporal regulation of GRF and bHLH74 expression during leaf development. Furthermore, a survey of miR396 sequences in different species showed variations in the 5\u2032 portion of the miRNA, a region known to be important for miRNA activity. Analysis of different miR396 variants in Arabidopsis thaliana revealed that they have an enhanced activity toward GRF transcription factors. The interaction between the GRF target site and miR396 has a bulge between positions 7 and 8 of the miRNA. Our data indicate that such bulge modulates the strength of the miR396-mediated repression and that this modulation is essential to shape the precise spatio-temporal pattern of GRF2 expression. The results show that ancient miRNAs can regulate conserved targets with varied efficiency in different species, and we further propose that they could acquire new targets whose control might also be biologically relevant.MicroRNAs (miRNAs) are \u223c21 nt small RNAs that regulate gene expression in animals and plants. They can be grouped into families comprising different genes encoding similar or identical mature miRNAs. Several miRNA families are deeply conserved in plant lineages and regulate key aspects of plant development, hormone signaling, and stress response. The ancient miRNA miR396 regulates conserved targets belonging to the Arabidopsis and other plants have revealed that they fulfill essential regulatory roles. Most of the ancient miRNAs regulate transcription factors involved in plant development and hormone signaling. Here, we characterize the miR396 regulatory network. While miR396 regulates GRF transcription factors, at least in angiosperms and gymnosperms, this miRNA additionally regulates another transcription factor of the bHLH class but only in Arabidopsis thaliana and closely related species. Most conspicuously, the regulation of both conserved and new targets is important for leaf development in Arabidopsis. We also show that miRNA variants can exist in certain species and that they can display an enhanced activity towards their targets. In summary, we propose that conserved miRNA regulatory networks might expand their functions by the recruitment of additional targets as well as by slight variations in the small RNA sequences.Plants and other multicellular organisms need precise spatio-temporal control of gene expression, and this regulatory capacity depends, in part, on small RNAs. MicroRNAs (miRNAs) are one class of \u223c21 nt small RNAs that originate from endogenous fold-back precursors found in plants and animals. They recognize complementary target sites in target mRNAs and guide them to cleavage or translational arrest. Studies of conserved miRNA networks in MicroRNAs (miRNAs) are small RNA molecules, \u223c21 nt in length, that are widespread regulators of gene expression in animals and plants , [3Sequence differences among related miRNAs could be important in plants as it has been shown for miR319 and miR159, which have very similar sequences but regulate different genes in vivo, confirming that a complex spectrum of miR396 sequences co-exists in nature , the monocot-specific version had only a minor effect (bHLH74 transcript levels were reduced only 25% in plants expressing the highest levels of the monocot-specific version (data not shown). Therefore, this monocot-specific version of miR396 is selectively more efficient towards the GRFs. The addition of an extra nucleotide to this variant causes a bulge to be formed on the miRNA side of the bHLH74/miR396 pair . Whole-mnsgenics . The pGRGRF2-GUS . The remGUS mRNA in two representative lines for each vector and observed that bulge removal from the GRF2 reporter caused approximately a two-fold reduction in GUS RNA, confirming its higher sensitivity to miR396 or short photoperiods (8 h light/16 h dark) at 23\u00b0C.To analyze the vein pattern, leaves were fixed with FAA and cleared with a chloral hydrate solution. Pictures were then taken under dark field illumination in a dissecting microscope. The number of branching points (NBP) GRF2 and bHLH74 was altered introducing synonymous mutations in a cloned wild-type genomic fragment using the QuikChange Site Directed Mutagenesis Kit (Stratagene). Artificial miRNAs MIM396 was generated by gene synthesis (Mr. Gene GmbH).See PROTEIN PHOSPHATASE 2A (AT1G13320) cDNA levels RNA was extracted using TRIzol reagent (Invitrogen) and 1,0 \u00b5g of total RNA was treated with RQ1 RNase-free Dnase (Promega). Next, first-strand cDNA synthesis was carried out using SuperScriptTM III Reverse Transcriptase (Invitrogen) with the appropriate primers. PCR reactions were performed in a Mastercycler ep realplex thermal cycler (Eppendorf) using SYBRGreen I (Roche) to monitor dsDNA synthesis. MiR396 and miR396_7-8insG levels were concurrently determined in each sample by stem-loop RT-qPCR To visualize reporter activity, transgenic plants were subjected to GUS staining, as described previously Small RNA sequences obtained from miRBase (17.0) A list of relevant AGI locus identifiers is provided in Figure S1bHLH74 homologs from several species. Alignment of partial coding sequences for bHLH74. A red box highlights the miR396 target site and a grey box depicts part of the coding sequence of the bHLH domain. Conserved positions across all species are indicated by asterisks. See Sequence alignment of (TIF)Click here for additional data file.Figure S2bHLH74 on Arabidopsis thaliana development. (A) Schematic representation of the 35S:bHLH74 and 35S:rbHLH74 constructs. (B) Phenotypes observed in 12-day old T1 seedlings overexpressing bHLH74 or rbHLH74. Phenotypes were classified according to their strength (numbers 1 to 4). Arrowheads indicate the elongated cotyledons observed only in 35S:rbHLH74 seedlings. Scale Bar: 2 mm. (C) Phenotype frequencies, according to panel (B), observed in at least 100 independent T1 plants expressing each vector.Effects of high expression levels of (TIF)Click here for additional data file.Figure S3Arabidopsis thaliana (88 miRNAs). (B) Plant mature sequences (miRBase 16.0) belonging to 42 species. A black bar in position 8 (highlighted with an asterisk) represents the contribution of the miR396 variants.Variations in the mature sequence of conserved miRNA families. (A) and (B) Variations in the mature sequence of miRNAs conserved in angiosperms (20 families). Bars represent the nucleotide changes in miRNA obtained for each family from position 1 to 21. Variation for each family was normalized to the number of members so that each family contributes equally. MiRNAs miR159 and miR319 were considered as a single family. (A) Variations in (TIF)Click here for additional data file.Figure S4Description of the method used to quantify miR396 variants. The retrotranscription reaction was performed using a stem-loop oligo that matches the three miR396 variants. For the qPCR, an equimolar mix of primers matching the miR396 variants was used. PCR efficiencies were checked to be equivalent for the different miRNAs.(TIF)Click here for additional data file.Figure S5Arabidopsis miR396b or miR396_7-8insG displaying an intermediate phenotype and transgenic plants expressing type see . A locke(TIF)Click here for additional data file.Figure S6bHLH74 target site with (A) Arabidopsis miR396a and (B) the monocot-specific variant (miR396_7-8insG).Interaction of the (TIF)Click here for additional data file.Figure S7Arabidopsis thaliana. (A) Scheme showing the secondary structure of the miRNA-miRNA* region in miR396 precursors from Arabidopsis thaliana, Selaginella moellendorffii and Picea glauca. miR396 sequence is indicated in red. Note that a G-A change in position 7 of the mature miRNA sequence (indicated in light gray) does not alter the secondary structure of the precursors. (B) and (C) Diagram showing the interaction between Arabidopsis GRF2 and miR396b (B) or the variant found in pine and poplar (C). (D) and (E) Phenotypes of independent transgenic seedlings overexpressing miR396b (D) or the miR396b_7A>G (E) variants. Phenotypes were classified as wild type, medium and strong which correspond to the first, third and fifth picture from the left in Overexpression of miR396_7A>G in (TIF)Click here for additional data file.Table S1Arabidopsis thaliana.Predicted targets of miR396 in (DOC)Click here for additional data file.Table S2Predicted targets of miR396 in poplar.(DOC)Click here for additional data file.Table S3Predicted targets of miR396 in rice.(DOC)Click here for additional data file.Table S4Sequences used to analyze the conservation of the miR396 target site.(DOC)Click here for additional data file.Table S5Expression of different miR396 variants in publicly available small RNA sequencing libraries.(DOC)Click here for additional data file.Table S6Binary plasmids prepared for this study.(DOC)Click here for additional data file.Table S7Relevant locus identifiers, mutant alleles and RT-qPCR primers.(DOC)Click here for additional data file."} +{"text": "Aspergillus nidulans (An) consists of the AnFus3 MAP kinase, the upstream kinases AnSte7 and AnSte11, and the AnSte50 adaptor. The fungal MAPK module controls the coordination of fungal development and secondary metabolite production. It lacks the membrane docking yeast Ste5 scaffold homolog; but, similar to yeast, the entire MAPK module's proteins interact with each other at the plasma membrane. AnFus3 is the only subunit with the potential to enter the nucleus from the nuclear envelope. AnFus3 interacts with the conserved nuclear transcription factor AnSte12 to initiate sexual development and phosphorylates VeA, which is a major regulatory protein required for sexual development and coordinated secondary metabolite production. Our data suggest that not only Fus3, but even the entire MAPK module complex of four physically interacting proteins, can migrate from plasma membrane to nuclear envelope.The sexual Fus3 MAP kinase module of yeast is highly conserved in eukaryotes and transmits external signals from the plasma membrane to the nucleus. We show here that the module of the filamentous fungus Aspergillus nidulans is able to release Fus3 as a complex from the membrane. Complexes of Fus3 can include two additional kinases and an adaptor protein, and these complexes can migrate from the membrane to the nuclear envelope where only A. nidulans Fus3 can enter the nucleus to control nuclear regulators. Revealing specific functions of cellular Aspergillus Fus3 complexes in signal transduction to control fungal development and secondary metabolism will be a fascinating future task.Mitogen activated protein (MAP) kinase cascades are conserved from yeast to man to transmit an external signal to the nucleus and induce an appropriate cellular response. The yeast Fus3 MAP kinase module represents a textbook paradigm for signal transduction. The pathway is activated by external sexual hormones triggering several kinases that transmit the signal at the plasma membrane to Fus3. Phosphorylated Fus3 is released from the membrane-associated module, crosses the cytoplasm, and enters the nucleus to activate transcription factors for sexual development. We describe here the Fus3 MAPK pathway of a filamentous fungus that controls sexual development as well as secondary metabolism, which are coordinated processes in filamentous fungi. Eukaryotic organisms communicate between cell surface and nucleus to respond to environmental signals. The mitogen-activated protein kinase (MAPK) module consisting of a cascade of three protein kinases represents a highly conserved eukaryotic signal transduction system present from yeast to man. MAP3K phosphorylates a second kinase, MAP2K which itself phosphorylates the MAPK. This final kinase phosphorylates nuclear target proteins to activate appropriate gene expression Saccharomyces cerevisiae represents a paradigm for signal transduction in eukaryotes The sexual pathway of the budding yeast Ste50 represents a second adaptor which binds to the Opy2 membrane anchor and provides membrane association of the entire MAPK module. Ste50 mediated membrane localization is required for Ste11 activation Aspergillus nidulans, the Ste11 MAP3K homolog SteC (AnSte11) A. nidulans grows vegetatively as a filament. When placed on a surface, after germination of the spores at least 12 hours of growth is required to establish developmental competence in response to external signals A. nidulans is coordinated with the production of secondary metabolites, including mycotoxins. This coordination requires velvet domain proteins which are common for filamentous fungi but absent in yeast Pheromone pathway genes have been studied in various fungi and are not only involved in sexual reproduction but also in fungal pathogenicity A. nidulans revealed that AnFus3 MAPK can reach the nuclear envelope in a complex with other proteins of the MAPK module, including the adaptor protein AnSte50. Only AnFus3 enters the nucleus and phosphorylates VeA, which elucidates a novel link between MAPK and velvet domain proteins that act as control elements at the interface of fungal development and secondary metabolism.Comparison of the intracellular molecular mechanism of signal transduction of Fus3 MAPK of yeast and S. cerevisiae Fus3 interacts with transcription factor Ste12 that activates the mating pathway. The A. nidulans MAP kinase AnFus3 [MpkB] also controls sexual development tandem affinity purification (TAP) only when the fungus was induced for sexual development but not during vegetative filamentous growth or asexual development resulted in loss of phosphorylation signal locus of A. nidulans encodes a protein, which is conserved in different Aspergilli and MAPK (MpkB) are necessary for sexual development in pergilli and has elopment \u2013S4C. mkkmkkB deletion strain was unable to fuse with the wild type strain. We found the same hyphal fusion defect for the steC\u0394 strain as in the mkkB mutant or with nuclear monomeric red fluorescent protein (mRFP) resulted in hyphae with green cytoplasm and red nuclei (heterokaryon) . In contB mutant . This fumkkB copy of the wild type strain allowed the development of mature fruiting bodies (red arrows), when wild type and mkkB mutant protoplasts were fused. In contrast, two mkkB mutants forced to form heterokaryons were impaired in fruiting body maturation and produced only early structures of development . Thus, the adaptor AnSte50 is as important for accurate fungal development as the other components of the MAPK module. A. nidulans AnSte50 was enriched by AnSte7::TAP in wild type, but not in the steC\u0394 strain indicating that AnSte11 is required for the AnSte50-Ste7 interaction and terrequinone (tdiA and tdiB), and the expression of laeA and the transcription factor encoding aflR, both required for expression of secondary metabolite genes, were distinctly reduced in each mutant of the MAPK module . After becoming competent for differentiation (16 hours after germination), AnSte7::sGFP accumulated not only at the hyphal tip but also at the plasma membrane and at the septa of hyphae or spore forming cells fication . Similarfication . There wfication . Quantiffication , suggestConsistently to the yeast situation, the transcription factor AnSte12 as well as fungus specific factors VeA and LaeA specifically interacted with the MAPK Fus3 in the nucleus . AnSte50A. nidulans is a homothallic fungus, which does not require a mating partner. Time lapse images revealed that MAPK module components AnSte7 and Ste50::sGFP can move within the fungal cell along the membrane. During the cellular movements, these molecules shortly touched the membrane then hit the nucleus. Sometimes, fusion protein moved back after contacting the nucleus in the opposite direction. . The AnSte7-Fus3 pair moved together along the plasma membrane . Contrastingly, the localization of the entire module at the hyphal tip or for the partial module AnSte11-Ste7 at the septum seems to be mediated by a mechanism which is largely independent of AnSte50.The interactions of the AnSte11-Ste7 and AnSte7-Fus3 complexes were examined in ild type was abolD mutant . Plasma A. nidulans MAPK module with the plasma membrane but it does not affect the hyphal tip and septum localizations.These data suggest that AnSte50 supports association of the A. nidulans Fus3 MAPK module which is involved in sexual development and the control of secondary metabolism and releases AnFus3 into the nucleus. Our data suggest a provocative additional hypothesis: AnFus3 is able to travel along the membrane and to cross the cytoplasm to the nuclear envelope in complexes with AnSte7 MAP2K, AnSte11 MAP3K and the adaptor protein AnSte50. In the nucleus AnFus3 interacts with transcription factor AnSte12 for sexual development. The additional interaction of AnFus3 with VeA or yet unidentifed targets may promote VeA-VelB formation which is required for coordinated development and secondary metabolism . This suggests a possible additional link between kinases of the module and regulators of asexual development.We describe here the tabolism . The A. A. nidulans. (i) From hyphal tip to plasma membrane: AnSte50 is primarily required for efficiently anchoring the MAPK module to membranes, but not to hyphal tips. AnSte50 might also contribute like in yeast to Ste11 MAP3K activation. The essential function of AnSte50 for signal transduction is supported by the defect of sexual development and lack of AnFus3 phosphorylation in a steD mutant. The AnSte50 independent localization at the hyphal tip suggests an additional yet unknown anchoring function for the AnFus3 module at the hyphal tip. The anchoring mechanism could include small membrane bound vesicles at the Spitzenk\u00f6rper which could explain some of our localization results In yeast Fus3 dissociates from the Ste5 tethered pheromone pathway module and enters into the nucleus (iii) Import of AnFus3 from nuclear envelope into nucleus: AnFus3 presumably dissociates from the kinase module at the nuclear envelope in a mechanism wihich is unknown. After entry into the nucleus, AnFus3 interacts with AnSte12, and presumably phosphorylates it. AnFus3 phosphorylates the velvet protein VeA, which efficiently associates with VelB and LaeA. It is yet unclear whether there are additional AnFus3 targets which support VelB-VeA complex formation. VelB-VeA then contributes with AnSte12 to sexual fruiting body development and the trimeric VelB-VeA-LaeA concomitantly promotes expression of distinct genes for secondary metabolites . These iA. nidulans is presumably involved in integrating multiple signals and enabling an adequate cellular response. Oxylipins represent currently the only known pheromones of Aspergilli but the receptors are unknown The MAPK module of A. nidulans could deepen insights into the molecular mechanism of information transfer through the cell.An interesting open question is whether other organisms also transport their Fus3 MAPK counterpart together with the entire module from surface to nuclear envelope. This results in questions about transport control points and module attachment sites on the nuclear envelope where future work in Aspergillus nidulans strains; FGSCA4 (veA+), TNO2A3 (veA1) veA+) veA+), AGB506 (veA+) veA+), AGB552 (veA+) served as wild type transformation hosts for the knock-out, epitope tagging, BIFC, and overexpression experiments. Further details for the strains transformed with various plasmids are given in nvitrogen) Escherichia coli strains were applied for recombinant plasmid DNA. Aspergillus and E. coli strains were cultured as described previously Fungal strains created and used in the course of this study are given in Pfu (MBI Fermentas), Phusion (Finnzymes), Platinum-Taq (Invitrogen) and Taq (Fermentas) were used. Linear and circular DNA constructs were created as given below.Circular and linear DNA molecules were created based on the standard recombinant DNA technology protocols in detail Anste7 [mkkB] deletion fragment, 1.1 kb 5 UTR and 0.6 kb 3 UTR flanking regions of AN3422 locus were amplified with 3422-A/C and 3422-D/F, respectively. These two fragments were fused with ptrA marker (amplified from pPTRII) by fusion PCR (3422-B/E), creating 3.6 kb deletion fragment which was transformed into TNO2A3 (veA1), AGB154 (veA+), creating AGB586, AGB587 strains, respectively. Complementation plasmid pME3854 was constructed by amplifying 4.2 kb genomic Anste7 [mkkB] locus (Comp-A/B) and subsequent cloning into StuI site of pAN8-1 For construction of Anste7 [mkkB] from sexual cDNA library PmeI site of pME3160 niiA promoter leading to pME3855 that was transformed into AGB152, which resulted in AGB662.Primers OZG302/303 amplified the 1.6 kb cDNA of Anste7 [mkkB] locus by mkkB::gfp and ctap, mkkB promoter including mkkB ORF (2.85 kb) and terminator regions (0.6 kb) were PCR-amplified from genomic DNA . Finally, the fragments 3422-A/OZG380 and OZG314/3422-F were fused to sgfp::natR module (with oligos 3422-B/3422E) creating 5.4 kb mkkB::sgfp::natR fusion construct. Likewise, 3422-A/OZG382 and OZG314/3422-F, and ctap::natR were joined by fusion PCR (3422-B/3422E) resulting in mkkB::ctap::natR cassette for gene replacement. mkkB::sgfp::natR construct (5.2 kb) was transformed into TNO2A3 and SWH51 mkkB::ctap::natR was introduced into AGB551 and SWH51 resulting in AGB597 and 598.For the purpose of substitution of the orinigal Anste11 [steC] ORF was amplified from gDNA (OZG389/OZG392) and fused to nyfp (OZG73/387) leading to nyfp::Anste11 [steC] fusion fragment which was cloned in PmeI site of pME3160 plasmid yielding pME3859 (nyfp::steC). Anste7 [mkkB] ORF was PCR-amplified from genomic DNA followed by fusion to nyfp and cyfp, which produced nyfp::Anste7 [mkkB] and cyfp::Anste7 [mkkB] fusions. Similar to Anste11 [steC] cloning, nyfp::mkkB fragment was inserted in PmeI site of pME3160 generating pME3861 plasmid.cyfp::mkkB was cloned in SwaI site of pME3859 leading to pME3860 that was brought in AGB506, which generated AGB599. For AnSte7/AnFus3 interactions, Anfus3 [mpkB] cDNA was amplified from cDNA library . OZG404/403 and OZG405/403 were fused to nyfp (OZG73/387) and cyfp (OZG75/388) fragments yielding nyfp::mpkB and cyfp::mpkB. nyfp::mpkB fragment was inserted in PmeI site of pME3160, which led to pME3864 and cyfp::mpkB was cloned in SwaI site of pME3861 generating pME3862 (nyfp::Anste7 [mkkB]/cyfp::Anfus3 [mpkB]). The BIFC plasmid pME3862 was introduced in AGB506 in order to generate AGB600. Anste12 [steA] gDNA was amplified with oligos OZG400/401 and fused to cyfp by fusion PCR (OZG75/400).To test AnSte11/AnSte7 interaction, cyfp::veA, cyfp::velB, cyfp::vosA and cyfp::laeA were produced as described in detail cyfp::steA, cyfp::veA, cyfp::velB, cyfp::vosA and cyfp::laeA in SwaI site of pME3864 yielded following plasmids pME3865 , pME3866 , pME3867 , pME3868 , pME3869 .Anste50 [steD] cDNA was amplified from cDNA library (OZG500/501) and joined to cyfp fragment with oligos OZG75/OZG500. Finally, this fragment was cloned in SwaI sites of pME3859, 3861, 3864, leading to plasmids pME3870, 3871 and 3927, respectively.steD deletion, steD::sgfp and steD::ctap linear fragments were created in an identical manner to mpkB constructs. steD deletion; OZG470/472, ptrA, OZG473/475 were fused by using oligos OZG471/474 in a fusion PCR. steD::sgfp::natR (5.1 kb); OZG470/564, sgfp::natR, OZG566/475 were joined by oligos OZG471/474. steD::ctap::natR (4.9 kb); OZG470/565, ctap::natR, OZG566/475 were joined by oligos OZG471/474. steD deletion cassette was brought into AGB552 and 551 generating, AGB576 and 650, respectively. steD::sgfp and steD::ctap were used for gene replacement in AGB551, giving rise to AGB657 and 659, respectively . OZG443/445, ptrA marker, and OZG446/448 were fused by oligos OZG444/447 creating 3.9 kb mpkB\u0394::ptrA construct. Consequently, mpkB\u0394::ptrA fragment was transformed into AGB552, which generated AGB611. To create Anfus3 [mpkB]::sgfp and ctap linear fragments, mpkB promoter as well as ORF was amplified by (OZG443/560 for gfp fusion and OZG443/561 for ctap fusion). OZG443/560, gfp::natR, and OZG562/448 were co-fused by oligos OZG444/447 creating 5.3 kb mpkB::sgfp::natR fragment. OZ443/561, ctap::natR, and OZG562/448 linear DNAs were fused to make mpkB::ctap::natR gene replacement fragment (5.1 kb). mpkB::sgfp and mpkB::ctap were transformed into the AGB551 strain, which yielded AGB654 and 659, respectively , mpkB cDNA (OZG737/738), and mRFP::H2A terminator (OZG739/740) were amplified and fused together (OZG735/740). The fusion was cloned in the SwaI site of a pyrG marker bearing plasmid. The final plasmid pME3966 was introduced into AGB590 and AGB657 for co-localization studies.ectively \u2013S9G. ForSTE7 and FUS3 were amplified from the wild type yeast genomic DNA. These regions were either fused to the gDNA or cDNAs of Anste7 [mkkB] (681/682) and Anfus3 [mpkB] (687/688) genes. Resulting fusions proSTE7::mkkB gDNAter::STE7, proSTE7::mkkB cDNAter::STE7, proFUS3::mpkB gDNAter::FUS3 and proFUS3:: mpkB cDNAter::FUS3 were cloned in SmaI site of yeast centromeric plasmid pRS316 (URA3) STE7 (OZG679/684), FUS3 (OZG685/690), and KSS1 (OZG691/692) genomic loci were amplified and similarly cloned into SmaI site of pRS316 resulting in pME3962, 3963, and 3964 plasmids, respectively. These control and chimeric constructs were transformed into the appropriate ste7, fus3, and fus3/kss1Promoter and terminator regions of Southern and Northern hybridizations were carried out as explained in detail ell Signaling Technology Inc) was used for detection of the phosphorylated AnFus3 [MpkB]. For the detection of the phosphorylated proteins, \u03b1-phosphoserine/threonine was employed. Manufacturers protocols were followed for incubation times and buffer applications of phosphospecific antibodies.Immunoblotting experiments for recognition of GFP, TAP fusion, VeA, and actin in protein extracts was performed according to described protocols ealthcare) and GST-tagged LaeA91 with a 5 ml GSH-Sepharose column connected to an \u00c4KTA Prime chromatography system. After washing with 10 column volumes with lysis buffer, proteins were eluted with elution buffer plus 400 mM Imidazol or 30 mM reduced Glutathione. Velvet proteins were desalted with a HiPrep Desalting 26/10 column into storage buffer . GST-LaeA91 was cleaved with PreScission Protease at 4\u00b0C for 16 h and further purified by gel-filtration using a Superdex 200 26/60 and a final 5 ml GSH-Sepharose column both equilibrated in gelfiltration buffer . All proteins were shock-frozen in liquid nitrogen and stored at \u221280\u00b0C until further use.Proteins were expressed in Rosetta 2 (DE3) using ZYM5052 hromotek) was washed twice with 1 ml protein extraction buffer . 20 ml protein crude extract was incubated with 100 \u00b5l GFP-Trap sepharose (Chromotek) at 4\u00b0C for 2 hours on a rotating platform. Afterwards, sepharose-extract mixture was centrifuged at 4000 rpm at 4\u00b0C for 1 min. Crude extract was removed with a 5 ml pipette. The sepharose was washed twice with 20 ml of protein buffer and centrifuged at 4000 rpm at 4\u00b0C for 1 min. This step was repeated one more time. Finally, 1 ml of protein buffer was added and the sepharose was resuspended. Each of the 200 \u00b5l sepharose buffer mixture was transferred into 1.5 ml eppendorf cups and centrifuged at 4000 rpm at 4\u00b0C for 1 min and supernatant was removed. Immunoprecipitated proteins were washed three times with 1 ml kinase reaction buffer .In order to immunoprecipitate GFP fusion proteins, protein crude extracts were prepared from vegetatively grown cultures. 100 \u00b5l GFP-Trap sepharose and incubated at 95\u00b0C for 10 min. 30 \u00b5l of the supernatant fraction was run on 4\u201315% gradient SDS gel that was dried for 2 h and exposed to Kodak X-omat film for 5 hours. 10 \u00b5l of the reaction was used for visualization of the proteins with coomassie staining. 2 \u00b5l of sepharose was used for immunoblotting and ponceau staing for validation of equal immunoprecipitated target protein (MpkB or GFP). For non-radioactive kinase experiments, same KRB buffer containing 5 \u00b5M ATP was used. Supernatants were treated with 1000 units lambda protein phosphatase (New England Biolabs) in the presence of 1 mM MnCl2 at 30\u00b0C for 1 hour. Samples were added with 3\u00d7 loading dye and boiled at 95\u00b0C for 10 min. 3 \u00b5l of the samples were used for immunoblotting.For the TAP purification of the MkkB, MpkB, SteD, and VeA interacting proteins and further LC-MS/MS identification previously published protocols were applied A. nidulans strains expressing various fluorescence proteins (EYFP/sGFP/mRFP) were inoculated in the 8-well borosilicate coverglass system (Nunc) containing the liquid minimal medium. Widefield fluorescence photographs were taken with an Axiovert Observer. Z1 (Zeiss) microscope equipped with a CoolSNAP ES2 (Photometrics) digital camera. CSU-X1 A1 confocal scanner unit (Yokogawa) connected with QuantEM:512SC (Photometrics) digital camera was used for laser microscopy. The SlideBook 5.0 software package (Intelligent Imaging Innovations) was used for fluorescence and laser confocal image and movie recording as well as productions. We defined signals as plasma-membrane localized if we found the signals that are at the border of the silhouette of the fungal cell or even surmount the fungal cell; similarly, we defined signals as nucleus-associated when we found multiple signals at the border of the nuclear silhouette.eica Microsystems), under 514 nm excitation (provided by an Argon laser), using a 100\u00d7 oil-immersion objective . The images were processed by a custom-written routine in Matlab (The Mathworks Inc.). Briefly, the spots were identified by the application of an automatic threshold based on the intensity of the background. We then used Gaussian fits to the spots to determine their intensity, and to correct for the background intensity, which provided the baseline of the fits.The EYFP protein was purified by using GFP-Trap as described for GFP protein. EYFP molecules were allowed to attach to poly-L-lysine coated coverslips for 10 minutes, in PBS buffer. Fungal cultures were grown as described above. The preparations were imaged using a SP5 TCS STED microscope (LExtraction of sterigmatocystin (ST) and thin layer chromatography (TLC) was carried out as given in detail Figure S1Anfus3 [mpkB\u0394], Anste50 [steD\u0394], Anfus3 [mpkB]::sgfp, Anfus3 [mpkB]::ctap, Anste50 [steD]::sgfp, Anste50 [steD]::ctap strains incubated under dark and light conditions for 5 days at 37\u00b0C. Black frames are the stereomicroscopic images of the colonies on the plates. Mature fruiting bodies are indicated by yellow arrows. Anfus3 [mpkB\u0394] and Anste50 [steD\u0394] strains cannot produce mature cleistotheica (indicated by red arrows) instead form nest-like structures. (B) Quantification of the asexual conidiations of the strains from (A). Reduced asexual sporulation seen in Anfus3 [mpkB] and Anste50 [steD] mutants. Replacement strains sporulate more efficient than the deletion strains. (C) Expression of AnSte50 [SteD] and AnFus3 [MpkB]::sGFP proteins during different developmental stages . 68 kDA AnFus3 [MpkB] and 81 kDA AnSte50 [SteD]::sGFP fusion proteins were detected by \u03b1-gfp. Constitutive transcript levels of the mpkB and steD genes from the same experiments. Actin levels served as loading control for immunoblotting (80 \u00b5g in each lane), and internal gpdA expression was used as control for Northern hybridizations. 20 \u00b5g RNA was loadeded in each sample.Functionality of AnFus3 [MpkB] and AnSte50 [SteD]::sGFP and TAP fusions for fungal growth, sexual development. (A) Phenotypes of the wild type, (TIF)Click here for additional data file.Figure S2velB and laeA and cellular localizations of the velvet complex components in the wild type and Anste7 [mkkB\u0394] strain. (A) Transcript levels of velB and laeA in the wild type and mpkB mutant background. RNA levels of velB and laeA from two different time points (24 and 72 hours) were quantified and normalized to the internal control gene expression gpdA. velB levels do not change significantly, but laeA transcript is drastically downregulated. Black bars represent standard deviations. (B) Localization patterns of VeA, VelB and LaeA::sGFP fusions in the wild type and mpkB mutant background. Fungal strains were grown in the darkness for 24 hours at 30\u00b0C and pictures were taken in a fluorescence microscope. Scale bars represent 10 \u00b5m.Transcript levels of (TIF)Click here for additional data file.Figure S3Aspergillus nidulans with other eukaryotic MAPK Kinase homologs. A. nidulans AnSte7 [MkkB] (ANID_03422) was aligned with the amino acid sequences from Aspergillus fumigatus Afu3g05900, Neurospora crassa MAPK Kinase (NCU04612), Magnaporthe oryzae (EHA48601.1), Podospora anserina (XP_001910826.1), Trichoderma atroviride (EHK42325.1), Coccidioides immitis (XP_001246770.1), Saccharomyces cerevisiae Ste7p, and Homo sapiens MAPK2K1. Conserved protein kinase domains and the central part showing higher similarity are indicated with red rectangle. N- and C-terminal sequences of the kinase proteins show less similarity. S. cerevisiae and H. sapiens proteins often break the alignment. Filamentous fungus kinases show higher similarity to the AnSte7 protein.Global alignment of the AnSte7 [MkkB] of (TIF)Click here for additional data file.Figure S4Anste7 [mkkB] gene and functional complementation assays in yeast. (A) Growth of the control (empty plasmid carrying strain) and Anste7 [mkkB] OE (under niiA promoter) strains under white light (90 \u00b5Wm2) and dark conditions on repressing (NH4+) and inducing (NO3\u2212) media. Lower panel shows the plate pictures, upper squares are the stereomicroscopic images taken from the plates. 1\u00d7104 spores were point-inoculated and grown at 37\u00b0C for 5 days. (B) Validation of Anste7 [mkkB] overexpression by Northern blot. gpdA transcript levels and rRNA were used as equal loading controls. Total 20 \u00b5g RNA was applied in each lane. (C) Quantification of cleistothecia production from (A). Increased cleistotheica production in Anste7 [mkkB] OE strain in the dark on nitrate containing inducing medium. Vertical lines are the standard errors originating from different counts. L; light, D; dark. Rep; repressed, Ind; induced. (D) Either cDNA or ORF of Anste7 [mkkB] and Anfus3 [mpkB] expressed under yeast STE7 or FUS3 promoters in a centromeric self-replicating plasmid. These constructs were expressed in the respective fus3, ste7 and fus3/kss1 double mutants. Strains were grown in the presence of 15 \u00b5g alpha factor given on the paper discs at 30\u00b0C for 3 days. Alpha factor in wild type (empty plasmid) and complementation strains results in a strong growth inhibition . ste7 and fus3/kss1 mutants do not show any response to the pheromone treatment. fus3 mutant exhibits a reduced response . AnSte7 and Fus3 do not remediate the halo phenotype of the ste7 and fus3 mutants. mpkB cDNA partially restores the pheromone response of the fus3/kss1 double mutant.Increased sexual development caused by overexpression of (TIF)Click here for additional data file.Figure S5Anste7 [mkkB]::sgfp, Anste7 [mkkB]::ctap, Anste7 [mkkB\u0394] strains on plates incubated under white light (90 \u00b5Wm2). Anste7 [mkkB]::sgfp and ctap look like the wild type strain. (B) Conidia production capacities of the strains from (A). Strains carrying Anste7 [mkkB]::sgfp and ctap constructs produce similar levels of conidia of the wild type levels. Vertical bars are the standard deviations of quantifications. Same spore number was used for inoculation as in A. nidulans. Strains were grown vegetatively 20 h, and transferred on plates and incubated under light conditions for asexual development and dark for sexual development. Protein undergoes degradation during 24 h asexual and sexual time points. Anste7 [mkkB] transcripts are expressed constitutively during different stages. Actin levels and ethidium bromide stained ribosomal RNA served as loading controls.Functionality of the AnSte7 [MkkB]::sGFP and cTAP tag fusions for fungal growth and sexual development. (A) Development of the wild type, (TIF)Click here for additional data file.Figure S6Quantification of the EYFP fluorescence intensities from single EYFP and AnSte11-Ste7 and AnSte7-Fus3 BIFC complexes. (A) The intensity of the spots produced by the AnSte11-Ste7 and AnSte7-Ste3 complexes were measured (see (TIF)Click here for additional data file.Figure S7Spatial movements of the AnSte7-AnFus3, AnSte50-AnFus3, AnSte11-AnSte7 binary complexes within the fungal cells. (A) Movement of the AnSte7 [MkkB]-AnFus3 [MpkB] complex (white arrow) that touches the nucleus during intracellular translocations (17 min) (Video S3). Yellow arrows indicate the direction of the movements. (B) A movement of the AnSte50-AnFus3 complexes between two nuclei. Complex leaves the first nucleus, and slightly touches the membrane reaches to the second nucleus (Video S4). (C) A horizontal backwards movement of the AnSte11 [SteC]-AnSte7 [MkkB] complexes from hyphal tip. Complexes touch the nucleus during bypass (Video S5). (D) A vertical movement of the AnSte11-AnSte7 complexes from membrane to the nuclear envelope (Video S6). White arrows indicate the YFP spots representing the binary complexes moving to the nucleus.(TIF)Click here for additional data file.Figure S8Anste7 [mkkB] locus. (A) A comparative depiction of the genomic Anste7 [mkkB] (AN3422) and the deletion of the locus by the selection marker pyrithiamine resistance gene, ptrA. Blue bar represents the Southern probe used in hybridizations. (B\u2013C) Southern hybridization results of the mkkB deletion and complementation strains. Sizes of the restriction bands confirm the gene replacement and ectopic complementation of the knock-out strain by the complementation plasmid. Sizes of the restriction fragments are given in base pairs. (D) Schematic drawings of the Anste7 [mkkB] locus gene replacements by Anste7 [mkkB]::sgfp::natR and Anste7 [mkkB]::ctap::natR. The cutting sites of the common restriction enzymes are indicated in the theoretical maps. (E\u2013F) Southern results of the Anste7 [mkkB]::sgfp::natR and Anste7 [mkkB]::ctap::natR strains in comparison to the wild type locus. Bands released by restriction digests are in agreement with the theoretical maps of the replaced loci.Southern hybridizations for the gene replacement experiments involving (TIF)Click here for additional data file.Figure S9Anste50 [steD] and Anfus3 [mpkB] loci. (A) Common restriction enzyme cutting maps of the wild type Anste50 [steD] (AN7252) locus, steD\u0394::ptrA, Anste50 [steD]::sgfp::natR, and Anste50 [steD]::ctap::natR gene replacements. Blue lines show the probe binding sites during Southern hybridizations. (B\u2013C) Southern hybridizations of gene replacements in comparison to the wild type Anste50 [steD] locus. Restriction enzymes used during Southern hybridizations are shown at the top of the blot. Lengths of the restriction fragments are given in base pairs. (D) Restriction map of the Anfus3 [mpkB] (AN3719) locus and corresponding gene replacements for deletion, sgfp and ctap epitope taggings. (E\u2013G) Southern results for Anfus3 [mpkB] gene replacements for sgfp, ctap and deletion. Bands produced by the restriction enzymes are compatible with the theoretical map of the Anfus3 [mpkB] locus. Blue bars indicate the regions where the Southern probes bind.Verification of the gene replacements for (TIF)Click here for additional data file.Table S1SEQUEST Multiple Consensus Report of AnFus3 [MpkB]::cTAP tag and sGFP identifications after nano-LC-ESI-MS2.(XLS)Click here for additional data file.Table S2SEQUEST Multiple Consensus Report of VeA::cTAP tag identifications in wild type after nano-LC-ESI-MS2.(XLS)Click here for additional data file.Table S3mpkB\u0394 strain after nano-LC-ESI-MS2.SEQUEST Multiple Consensus Report of VeA::cTAP tag identifications in (XLS)Click here for additional data file.Table S4SEQUEST Multiple Consensus Report of AnSte7 [MkkB]::cTAP tag identifications after nano-LC-ESI-MS2.(XLS)Click here for additional data file.Table S5steC\u0394 strain after nano-LC-ESI-MS2.SEQUEST Multiple Consensus Report of AnSte7 [MkkB]::cTAP tag identifications in (XLS)Click here for additional data file.Table S6SEQUEST Multiple Consensus Report of AnSte50 [SteD]::cTAP tag identifications after nano-LC-ESI-MS2.(XLS)Click here for additional data file.Table S7Fungal strains used in this study.(DOC)Click here for additional data file.Table S8Plasmids employed in this study.(DOC)Click here for additional data file.Table S9Oligonucleotides utilized for plasmid constructions and northern hybridizations.(DOC)Click here for additional data file.Video S1Time-lapse analysis of the subcellular movements of the AnSte7-GFP fusion along the fungal cells. Individual focal planes were captured with a spinning disc confocal microscope at 2 min intervals . AnSte7 protein moves in an internuclear manner. The nuclei were visualized by mRFP::Histone2A fusion protein. Green spot leaves the first nucleus and shortly touches the plasma membrane (zigzag movement) and sticks to the envelope of the next nucleus. Some spots of the AnSte7 are static . The video is presented at a rate of 5 frames/second.(MOV)Click here for additional data file.Video S2Time-lapse capture of the subcellular movements of the AnSte50-GFP fusion within the fungal cell. Single focal planes were captured at 2 min intervals . AnSte50 protein moves to the nucleus (red), hits the nuclear envelope or nucleus and moves in a retrograde direction. Due to the single focal plane, spot disappears at 6 min. After 6 min, spot movement can be tracked again. The video speed is 5 frames/second.(MOV)Click here for additional data file.Video S3Time-lapse analysis of the subcellular movements of the AnSte7-AnFus3 complexes (yellow dot) along the fungal hypha. AnSte7-Fus3 complexes move in a retro and anterograde direction in comparison to the hyphal tip (upper left). These complexes move between the nuclei, which were visualized by mRFP::Histone2A fusion. Single focal layer images were captured at 1 min intervals . The video was produced at a setting 5 frames/second.(MOV)Click here for additional data file.Video S4Time-lapse analysis of the subcellular dynamics of the AnSte50-AnFus3 complexes (yellow dot) in the fungal hypha. Single focal pictures were taken at 2 min intervals . This movie shows the movement of the AnSte50-AnFus3 complexes between two nuclei. Protein complexes (yellow dot) leave the first nucleus and move to the second one. While moving to the second one, the complexes slightly touch the plasma membrane. Video is presented by using the setting 5 frames/second.(MOV)Click here for additional data file.Video S5Retrograde translocation of the AnSte11-Ste7 protein complexes along the fungal hypha. AnSte11-Ste7 complexes move backwards from the hyphal tip. They leave the membrane and touch the nucleus. They also accumulate at septa (faint immobile yellow dot). Single focal planes were captured at 2 min interval . The video is presented at the speed of 5 frames/second.(MOV)Click here for additional data file.Video S6Time-lapse analysis of the migration of AnSte11-Ste7 complexes from the plasma membrane to the nucleus. Membrane-tethered complexes (yellow spots) slowly move to the nucleus (red). The movie was captured at 1 min interval . 5 frames/second.(MOV)Click here for additional data file."} +{"text": "We conclude that PTN delays the maturation of the mammary gland by maintaining mammary epithelial cells in a progenitor phenotype and by inhibiting their differentiation during mammary gland development.Expression of the heparin-binding growth factor, pleiotrophin (PTN) in the mammary gland has been reported but its function during mammary gland development is not known. We examined the expression of PTN and its receptor ALK (Anaplastic Lymphoma Kinase) at various stages of mouse mammary gland development and found that their expression in epithelial cells is regulated in parallel during pregnancy. A 30-fold downregulation of PTN mRNA expression was observed during mid-pregnancy when the mammary gland undergoes lobular-alveolar differentiation. After weaning of pups, PTN expression was restored although baseline expression of PTN was reduced significantly in mammary glands of mice that had undergone multiple pregnancies. We found PTN expressed in epithelial cells of the mammary gland and thus used a monoclonal anti-PTN blocking antibody to elucidate its function in cultured mammary epithelial cells (MECs) as well as during gland development. Real-time impedance monitoring of MECs growth, migration and invasion during anti-PTN blocking antibody treatment showed that MECs motility and invasion but not proliferation depend on the activity of endogenous PTN. Increased number of mammospheres with laminin deposition after anti-PTN blocking antibody treatment of MECs in 3D culture and expression of progenitor markers suggest that the endogenously expressed PTN inhibits the expansion and differentiation of epithelial progenitor cells by disrupting cell-matrix adhesion. In The heparin-binding growth factor pleiotrophin start undergoing lobular-alveolar differentiation. We also found that blocking PTN activity caused enhanced maturation of the mammary gland accompanied by activation of the ERK1/2 signaling pathway in the epithelial compartment of the mammary gland. We show that PTN activity sustains motility and invasion of MECs grown on plastic and that blocking PTN activity caused increased number of mammospheres due to a more polarized structural organization shown by laminin deposition and a more differentiated phenotype as indicated by the expression of progenitor cell markers CD29, CD49f, SCA-1 and CD10.in situ hybridization as well as cell fractionation followed by Northern blot . This suggests that sufficient dosing was used to block endogenous PTN. Mice treated during the crucial phase of mammary duct invasion into the fat pad from 3 to 6 weeks of age showed an increase in ductal penetration of the mammary fat pad and in the average number of branch points per duct PTN is expressed and highly regulated during embryonic and postnatal development of different tissues The mammary gland is one of the few organs that mature after birth and continues development during puberty under hormonal stimuli until adulthood when it becomes quiescent. During pregnancy, the mammary gland goes through several changes such as proliferation, differentiation and apoptosis of the alveolar cells. The multiple cell populations in the mammary gland change in number and ratio during the process of development as well as during pregnancy. Using different, independent approaches we show here that PTN is predominately expressed in epithelial cells in the virgin mammary gland. The tissue analysis was also validated in isolated mammary epithelial cells (MECs) harvested in parallel with isolated fibroblasts showing a >10 fold higher expression of PTN in the MECs than in fibroblasts and exprPTN expression remained unaltered during the first 10 days of pregnancy but was downregulated 30-fold during late pregnancy when the mammary gland is proliferating and differentiating in preparation for lactation. A downregulation of PTN expression during late pregnancy was surprising to us because the epithelial ducts are still proliferating and PTN has previously been shown to be a growth factor for different types of cells There is only one known PTN homologue, midkine (MK), that shares 50% protein identity PTN was found to be a ligand that binds to the extracellular domain of the tyrosine kinase receptor ALK When comparing PTN expression among nulliparous, uniparous, multiparous and pseudopregnant mice, a significant decrease in PTN mRNA expression level was detected in the mammary gland of multiparous versus nulliparous mice but not in pseudopregnant mice. These findings suggest a regulation of PTN expression due to gestational hormones and a decrease in PTN expression possibly due to a loss in mammary epithelial progenitor cells by repeated pregnancies. Full term pregnancy has been demonstrated as a protective factor against the risk of breast cancer PTN mRNA expression in the mammary epithelial cells has been previously suggested The fact that the anti-PTN antibody treated MECs formed a higher number of mammospheres with correct apical polarity and laminin basement membrane deposition than control MECs in 3D cultures suggests that PTN inhibits polarization of MECs. PTN expression has been shown in neuronal progenitor cells and its activity enhances progenitor cells expansion and differentiation in different tissues PTN appears to inhibit differentiation of mammary epithelial cells and maintain their progenitor phenotype that is evidenced during mammary gland development as well as in isolated mammary epithelial cells in culture. It is tempting to speculate that PTN downregulation observed with multiparity is protective towards breast cancer, possibly by reducing the immature precursor population susceptible to malignant transformation and by expanding differentiated epithelial cells.All procedures using animals were approved by the Georgetown University IACUC and were conducted according to the NIH guidelines for the care and use of laboratory animals.C57BL/6, Balb/c and FVB mice were purchased from the Jackson Laboratory. All procedures using animals were approved by the Georgetown University IACUC and were conducted according to the NIH guidelines for the care and use of laboratory animals.Mammary glands or brains were isolated from female balb/c mice. Involution days are given relative to the weaning of 21 day old pups with mammary glands from exbreeders harvested >4 weeks after weaning. All experiments were performed using combined abdominal and thoracic mammary glands. Tissues were homogenized in RNA-STAT, and the RNA was isolated via a standard chloroform extraction procedure. For experiments requiring tissue fractionation, mammary glands were finely minced at 4\u00b0C and then incubated in DMEM Hams F12 media supplemented with 0.15% collagenase for 1 hour at 37\u00b0C. After collagenase digestion, glands were passed through cheesecloth (Baxter) and filtered through a cell strainer using a 70 \u03bcm mesh .The filtrate was centrifuged for 5 minutes at 500\u00d7g . Both the filtrate pellet and the filtered collected fraction were resuspended in RNA-STAT and processed for RNA extraction as described above.32 labeled probes primed from full length cDNA for mouse Keratin 18 TM gene expression was quantified using SYBR green (BioRad). Primers were:Northern blots were performed as previously described TGGAGAATGGCAGTGGAGTGTGTPTN (Forward) TGGTACTTGCACTCAGCTCCAAACTPTN (Reverse) CAAGTCTGCCGAAATCAGGGACGK18 (Forward) TCCAAGTTGATGTTCTGGTTTTTCATGGK18 (Reverse) GGACGGGACACAGCTCCATGALK (Forward) GCACTCCAGACCATATCGACTGCGALK (Reverse) CTCTCTGTGCTTGTCTTGCTCCD10 (Forward) GACGTTGCGTTTCAACCAGCCD10 (Reverse) ATGCCAAATCTTGCGGAGAATCD29 (Forward) TTTGCTGCGATTGGTGACATTCD29 (Reverse) TGCAGAGGGCGAACAGAACCD49f (Forward) GCACACGTCACCACTTTGCCD49f (Reverse) AGGAGGCAGCAGTTATTGTGGSCA-1 (Forward) CGTTGACCTTAGTACCCAGGASCA-1 (Reverse) Primers for Vimentin were described in Abdominal mammary glands were fixed in formaldehyde and embedded in paraffin. Immunohistochemical analysis of 5 \u03bcm section was done using the Vectastain ABC Elite System (Vector Labs). The primary goat anti-human PTN antibody (R&D) diluted 1\u2236500 was incubated at 4\u00b0C overnight. The slides were counterstained with hematoxylin (Sigma Aldrich). Tissue sections are representative of 3\u20134 mice. Image capturing for stained section was performed with a Nikon E600 Epifluorescence Microscope.Mammary gland #4 was excised from 6-week-old mice whole mount stained D-value of 1 nM.The hybridoma cell line producing the anti-PTN mouse monoclonal antibody (clone 3B10) was previously described FVB mice (3 weeks old) were injected intraperitoneally with a total dose of 2 mg/kg of the antibody or with PBS as a control. Serum samples from control and treated 6-week-old FVB mice were diluted 1\u2236100 in PBS and tested in an ELISA assay with immobilized recombinant human Pleiotrophin . 5% (w/v) non-fat dry milk was used to block non-specific binding. Superfluous solution was washed off with 3 washes with 0.05% Tween20 (Fisher Scientific) in PBS (\u200a=\u200aPBST). Diluted serum samples were incubated for 1 h at room temperature. Anti-PTN antibody stock solution diluted to 100 nM was used for calibration. Wells were washed 5 times with PBST. A horseradish peroxidase-linked sheep, anti-mouse IgG was the secondary antibody , 1-Step Turbo TMB ELISA Substrate (Thermo Scientific) the detection reagent and sulfuric acid the stop solution. The absorbance was read at 450 nm.Whole mammary glands #3 from 6-week-old FVB mice was harvested and lysates (5 \u00b5g/well) were analyzed either using a MS6000 MAP Kinase Whole Cell Lysate kit (Phospho protein) and MS6000 MAP Kinase Whole Cell Lysate kit according to manufacturer's instructions. Whole mammary gland lysates and whole cell lysates from primary cultures were run (75 \u00b5g/lane) on a 4\u201320% Tris-Glycine NOVEX gel (Invitrogen), transferred to PVDF membranes. Primary epithelial cells were harvested 24 hours after treatment with UO126 (500 nM) alone or in combination with anti-PTN blocking antibody. Immunoblot studies for phospho-MAPKs and total MAPKs were performed with the respective rabbit polyclonal antibodies . Visualization was performed using Immobilon Western HRP substrate Peroxide Solution (Millipore) with horseradish peroxidase-linked sheep anti-rabbit immunoglobulin G as a secondary antibody .2 and cultured up to passage #3 and then used for RNA extraction, immunofluorescence, three-dimensional culture, real-time impedance sensing and Western blot analysis.Mammary glands #3 and #4 were dissected from 7 to 9-week-old FVB mice carefully removing muscles, lymph nodes and nipples. Glands were minced and digested in 1 mg/ml collagenase (Sigma-Aldrich) solution for 90 minutes at 37\u00b0C on a shaker at 150 rpm. The digested tissue was centrifuged for 10 minutes at 600\u00d7g and washed 4 times. DMEM/F12 (Invitrogen) media supplemented with Penicillin/Streptomicin (Invitrogen) 1X and 50 \u00b5g/ml Gentamicin (Invitrogen) was used to prepare the collagenase solution and to wash the digested tissues. Cells were resuspended and plated on collagen-coated plates (Costar) in MEC growth media (Ham's F12 (Invitrogen) supplemented media with 10% fetal bovine serum, 16 \u00b5g/ml insulin (Invitrogen), 10 ng/ml EGF (Roche Applied Biosciences), 1 \u00b5g/ml hydrocortisone (Sigma-Aldrich), 50 \u00b5g/ml gentamicin (Invitrogen), 4 ng/ml cholera toxin (Sigma-Aldrich) and 1X penicillin/streptomycin). To avoid fibroblast contamination, cells in the supernatant were transferred to uncoated plates 20 minutes after initial plating. Fibroblasts growing on collagen coated plates (BD Biosciences) and epithelial cells were incubated at 37\u00b0C and 5%COPrimary epithelial cells were plated on microelectrodes embedded wells and the impedance of the cell monolayer was monitored in real time as described elsewhere 3D culture was performed as described in Primary mammary epithelial cells passage #3 grown in poly-D-lysine coated 8 well Lab-Tek II Chamber Slide System (Nunc) were fixed and permeabilized twice with 4% formaldehyde, 0.2% Triton X in cold PBS for 10 minutes at room temperature, washed with ice-cold PBS, blocked with 1% BSA in PBS and incubated with a goat anti-human PTN antibody for 1h at room temperature. After three washings with PBS, cells were incubated with Alexa Fluor 488 anti -goat IgG and with Alexa Fluor 568 Phalloidin for 1h at room temperature. After three washings with PBS, cover slips were mounted with ProLong Gold Antifade with DAPI (Invitrogen) and fluorescence was analyzed on an Olympus Fluoview FV300 Laser Confocal Microscope .4Cl was added to samples for 5 minutes. Samples were washed 3 times with PBS and then immunostained as described in Immunostaining of mammospheres was performed as described in Prism 5 for Mac was used for all statistical analyses. Adobe Photoshop CS3 was used to process the pictures for immunohistochemistry, whole mounts, Western blot, mammosphere three-dimensional culture and immunofluorescence. Brightness/Contrasts and Levels adjustments were applied to the whole image. All images were adjusted to the same level. Image J was used to measure the ductal length, terminal end buds and terminal ends diameter, and mammosphere surface area. Immunofluorescence images were overlaid using the Metamorph Software.This article contains supporting information.Figure S1Anti-PTN antibody binding activity to PTN and its detection from mouse serum of treated mice. (A) Anti-PTN antibody binding to immobilized PTN by ELISA. Anti-PTN antibody KD-value \u200a=\u200a1 nM. Control\u200a=\u200a antibody elution buffer. OD defines absorbance at 450 nm. (B) PTN blocking antibody detection from mouse serum of treated mice versus control by PTN immobilized ELISA. Absorbance at 450 nm is defined as arb.u. Data are means \u00b1 SE; ***P\u200a=\u200a0.0004; two-tailed student-t test.(TIF)Click here for additional data file."} +{"text": "To analyze left ventricular (LV) remodeling using an accurate 3D evaluation of afterload-related changes in LV geometry.To maintain an effective LV-arterial coupling the LV adapts to the increased afterload caused by aging or cardiovascular disease. However, subsequent changes in LV mass and concentric remodeling have been associated with poor outcome. To understand LV remodeling, we studied variations of 3D myocardial wall stress (MWS), its geometrical factor as well as diastolic LV mass to volume ratio (LVM/EDV) on a population with a wide range of afterload.2). All subjects had short axis cine CMR acquisitions (GE 1.5T) followed by carotid applanation tonometry calibrated using brachial pressures recorded during CMR. After myocardial delineation, the LV geometrical factor LVGF, previously described by Grossman, was calculated as a combination of the local LV radius and myocardial thickness while considering the LV longitudinal curvature to correct for partial volume effects, especially in apical slices. This LVGF was combined with peak systolic pressures (PSP) resulting in MWS. For AVS patients, the echocardiographic transaortic valve maximal gradient was added to the tonometric PSP.Indeed, we studied 57 patients divided into three subgroups: 1) C1 included 22 healthy subjects aged between 22 and 37 years (26\u00b15 years), (2) C2 included 23 healthy subjects aged between 41 et 81 years (55\u00b19 years) and 3) AVS included 12 subjects (75\u00b114 years) with aortic valve stenosis (AVS) characterized by while LVM/EDV was increased in AVS significantly but failed to discriminate between age groups without AVS. Gradual changes in LV geometry reflected by the 3D LVGF demonstrated the ability of the LV to adapt to the increased afterload and therefore to maintain constant MWS. Indeed, no significant variations in MWS were found between the three subgroups Ejection fraction was homogeneous between the three subgroups The significant and gradual elevation in PSP found between the three groups caused changes in LV geometry (figure The described 3D evaluation of LV geometry, which can be easily integrated to standard CMR LV function evaluation since it only requires routine myocardial delineation in systole, sensitively characterized LV remodeling related to aging or to AVS."} +{"text": "XRCC1 (Arg399Gln), OGG1 (Ser326Cys), XRCC3 (Thr241Met), and XRCC4 (Ile401Thr) with the level of DNA damage and repair, accessed by comet and micronucleus test, in 51 COPD patients and 51 controls.We investigated a potential link between genetic polymorphisms in genes Peripheral blood was used to perform the alkaline and neutral comet assay; and genetic polymorphisms by PCR/RFLP. To assess the susceptibility to exogenous DNA damage, the cells were treated with methyl methanesulphonate for 1-h or 3-h. After 3-h treatment the % residual damage was calculated assuming the value of 1-h treatment as 100%. The cytogenetic damage was evaluated by buccal micronucleus cytome assay (BMCyt).XRCC1 (Arg399Gln) and XRCC3 (Thr241Met) showed higher DNA damage by comet assay. The residual damage was higher for COPD with risk allele in the four genes. In COPD patients was showed negative correlation between BMCyt with pulmonary function and some variant genotypes.COPD patients with the risk allele XRCC1 (Arg399Gln), OGG1 (Ser326Cys), XRCC3 (Thr241Met), and XRCC4 (Ile401Thr), DNA damage and progression of COPD.Our results suggest a possible association between variant genotypes in Recently genetic association studies on chronic obstructive pulmonary disease (COPD) risk have focused on identifying effects of single nucleotide polymorphisms/haplotypes in candidate genes, among which DNA repair genes are increasingly studied because of their critical role in maintaining genome integrity . Assays OGG1), apurinic/apyrimidinic endonuclease (APE1/APEX1), and the X-ray repair cross complementing group 1 (XRCC1). OGG1 and XRCC1 play a central role in the BER pathway. OGG1 catalyzes the removal of 8-hydrodeoxy-guanine (8-OHdG), which has been considered as a key biomarker of oxidative DNA damage. The substitution of cysteine for serine at codon 326 (Ser326Cys) is associated with a significant reduction in the repair capacity and the completion of recombination events) [XRCC1 (Arg399Gln) and OGG1 (Ser326Cys) are suggested to exert combined effect on the development of COPD (i.e. among current/light smokers), and XRCC1 coordinates and stimulates the OGG1 activity [Our results detected increased basal DNA damage in COPD patients with genetic polymorphisms XRCC1 Arg99Gln andquencies as well quencies . An assopulation . Geneticge assay . DNA strge assay . Thus, gge assay . In contge assay found no events) . Also, XGG1 Ser32Cys are sactivity ,45.XRCC1 (Arg399Gln), OGG1 (Ser326Cys), XRCC3 (Thr241Met), and XRCC4 (Ile401Thr) genetic polymorphisms on DNA repair was found in COPD patients compared to controls, as shown by residual damage Table\u00a0. This isWith our results, it is possible to assume an increase in the cancer risk analyzing the correlations that we found between pulmonary function and BMCyt because elevated binucleated cell ratio may be an indicative of higher aneuploidy rate which, in turn, is associated with an increased risk of cancer. The mechanism triggering nuclear bud formation is unknown but may be related to the elimination of amplified DNA or DNA repair complexes ,52,53. SHowever, there are some factors that should be taken into consideration, as the limited database. The sample size may not have been large enough to detect certain differences and there are other genetic polymorphisms that could also influence the repair capacity and consequently the risk for COPD and other repair pathways could also influence repair efficiency.XRCC1 (Arg399Gln) and XRCC3 (Thr241Met). In addition, the induced DNA damage by MMS increases in COPD patients with variant genotypes in XRCC1 (Arg399Gln), OGG1 (Ser326Cys), XRCC3 (Thr241Met) and XRCC4 (Ile401Thr) showing impairment of DNA repair. Some variant genotypes seem to be related to an increase in Bi, BUD, CC, KR cells, suggesting their possible influence on the background levels of this cytogenetic biomarker and progression of COPD.Finally, this investigation suggests that basal DNA damage increases, analyzed by Comet and BMCyt assays, in COPD patients with variant genotypes in The authors declare that they have no competing interests.ALGdS, DJM, ARdMV and JAPH prepared the draft of the manuscript. All the authors collected the data and contributed to the interpretation of results and to the revision of the manuscript. All the authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2350/14/93/prepub"} +{"text": "Although pre-existing heart disease in children is the most frequent predisposing factor for infective endocarditis (IE), we reporte 6 children of IE with no apparent pre-existing cardiac disease.-1ml-1) in children with IE and in IE-free children . For determination of PChE activity vein blood samples were collected and stored at -20\u00b0C until analyzed. PChE activity was determined by the spectrophotometric method of Ellman using butyryltiocholine as the substract . Statistical analysis was made by Student\u2019s t-test.Our study included six patients with acquired infection from central catheter; the microbial pathogens were Streptococcus viridans (n=3) and Staphyloccocus aureus (n=3). The diagnosis of IE was based on positive blood culture (2 positive cultures of blood samples drawn > 12 hours), evidence of endocardial involvement (positive ECHO), measuring raised erythrocyte sedimentation rate and plasma cholinesterase (PChE) activity. We measured PChE activity relation to patients of control group B .Our study indicates that PChE activity can be also an important parameter for the diagnosis of IE."} +{"text": "In last decades, obesity became a major health issue worldwide. Processes as well as molecules involved in fat mass regulation remain unresolved up to now. Brown adipose tissue (BAT) is an interesting target to fight obesity, especially since brown adipose tissue in adult humans was recently \u201crediscovered\u201d. In addition, the occurrence of \u201cbrown-like\u201d adipocytes within white adipose tissue (WAT) has also sparked an interest due to potential anti-obese effect. Transforming growth factor-\u03b2 (TGF-\u03b2) superfamily consists of structurally related but functionally diverse cytokines involved in regulation of adipogenesis. Activin receptor-like kinase-7 (Alk7), a member of the TGF-\u00df family, is highly expressed in adipose tissue. However, its role in adipogenesis, thermogenesis and adipocyte function is not well known.via activation of PKGI, since PKGI KO adipocytes show reduced levels of Alk7. Moreover, our preliminary data suggest involvement of PKGI in Alk7 promoter activation.Here, we analyzed regulation of both brown and white adipocytes focusing on Alk7 pathways. Levels of Alk7 mRNA are almost negligible at the beginning of differentiation protocol. Interestingly, expression of Alk7 increases with differentiation of brown and white adipocytes and reaches highest expression levels close to the end of differentiation protocol. Chronic treatment with 8-pCPT-cGMP (200 \u00b5M) increases expression of Alk7 in both brown and white adipocytes. Expression of Alk7 mRNA by cGMP is regulated Presently we are investigating effects of Alk7 activation in the presence and absence of cGMP onto adipocyte differentiation and BAT function.Taken together, our data show that cGMP via PKGI modulates expression of Alk7 in both white and brown adipocytes, thus pointing towards Alk7 as a new PKGI target.Considering expression pattern of Alk7, this interaction could prove beneficial for developing new pharmacological treatments to fight obesity."} +{"text": "Studies in HEK293T cell culture, murine skeletal muscle explants and ELISA of tissue homogenates showed efficacy of constructed plasmids. Functional activity of angiogenic proteins secreted by HEK293T after transfection by induction of tube formation in human umbilical vein endothelial cell (HUVEC) culture. HUVEC cells were used for in vitro experiments to assay the putative signaling pathways to be responsible for combined administration effect one of which could be the ERK1/2 pathway. In vivo tests of VEGF165 and HGF genes co-transfer were conceived in mouse model of hind limb ischemia. Intramuscular administration of plasmid encoding either VEGF165 or HGF gene resulted in increased perfusion compared to empty vector administration. Mice injected with a mixture of two plasmids (VEGF165+HGF) showed significant increase in perfusion compared to single plasmid injection. These findings were supported by increased CD31+ capillary and SMA+ vessel density in animals that received combined VEGF165 and HGF gene therapy compared to single gene therapy. Results of the study suggest that co-transfer of VEGF and HGF genes renders a robust angiogenic effect in ischemic skeletal muscle and may present interest as a potential therapeutic combination for treatment of ischemic disorders.Increased interest in development of combined gene therapy emerges from results of recent clinical trials that indicate good safety yet unexpected low efficacy of \u201csingle-gene\u201d administration. Multiple studies showed that vascular endothelial growth factor 165 aminoacid form (VEGF165) and hepatocyte growth factor (HGF) can be used for induction of angiogenesis in ischemic myocardium and skeletal muscle. Gene transfer system composed of a novel cytomegalovirus-based (CMV) plasmid vector and codon-optimized human VEGF165 and HGF genes combined with intramuscular low-voltage electroporation was developed and tested Therapeutic angiogenesis using plasmid vectors carrying growth factor gene is a broadly studied approach which has already entered clinical trials and remained an important field of translation research in recent decades. For example angiogenic potential of plasmids with VEGF, HGF and basic fibroblast growth factor has been shown previously Human studies showed safety of local administration of plasmid DNA Finally it is possible to enhance angiogenic response using gene therapy with physiologically additive combinations of factors. Published animal studies report increase of efficacy after gene therapy with pairs of VEGF/angiopoietin-1 in vitro findings of increased angiogenic response induced by combination of these two factors in cell culture and non-ischemic animal models of angiogenesis Among other growth factors a pair of VEGF and HGF is a potential candidate for therapeutic application. Both proteins are known to show morphogenic, mytogenic and antiapoptotic properties which are crucial for their regenerative potential Basing on these findings we hypothesized that combined transfer of VEGF and HGF genes to ischemic tissue could be a potent way to induce revascularization and restore blood flow. To demonstrate that we used a novel highly effective plasmid vector pC4W previously produced and described by our group All animal experiments carried out for the study were conducted in accordance with Institute of Experimental Cardiology guidelines. Animal procedures were approved by the Ethics Board of Institutional Animal Care and Use committee of Cardiology Research Complex (permit number 385.06.2009).Dulbecco's modified Eagle medium (DMEM), M199 medium with Earle's salt, 100x antibiotic/antimycotic solution were purchased from Gibco, fetal bovine serum was purchased from HyClone. Growth-factor reduced Matrigel basement membrane matrix (Cat#354635), recombinant human VEGF165 (Cat#354107) and HGF (Cat#354103) were purchased from BD Biosciences.EndoFree plasmid Giga kit (Cat#12391) was purchased from Qiagen.human Quantikine\u00ae VEGF165 (DVE00) and HGF (DHG00) kits were purchased from R&D Systems.mouse anti human VEGF165 monoclonal antibodies (Cat#MAB3045), mouse anti human HGF monoclonal antibodies (Cat#MAB294) were purchased from R&D Systems; Pharmingen rat anti mouse PECAM (CD31) monoclonal antibodies (Cat#550274) were purchased from BD Biosciences. FITC-labeled mouse polyclonal anti smooth muscle antibodies (Cat#F3777) were purchased from Sigma-Aldrich; AlexaFluor\u00ae594-conjugated donkey anti rat secondary antibodies (A21209) were purchased from Invitrogen; western blotting HRP-conjugated goat anti-mouse secondary antibodies (Cat#115-035-174) were purchased from Jackson Immunoresearch.Protease inhibitor cocktail (Cat#P-2714) was purchased from Sigma-Aldrich; 1x Quick Start Bradford Dye Reagent was purchased from Bio-Rad (Cat#500-0205);Human embryonic kidney (HEK) 293T cells were obtained from ATCC; human umbilical vein endothelial cells (HUVEC) were kindly provided by prof. A.V. Mazurov DSc from Laboratory of cell adhesion of Institute of Experimental Cardiology ex vivo detection of transgene expression and electrotransfer efficacy evaluation with \u03b2-gal reporter plasmid. All animals received standard food and water ratios. Surgical manipulations and euthanasia protocols were designed in accordance to Institute and National regulations.Eight-week old C57BL/6 male mice were used for hind limb ischemia experiments, E. Coli \u03b2-galactosidase gene sequence into pC4W vector.Mammalian expression vector pC4W as well as codon-optimized human VEGF165 and HGF genes have been described earlier Escherichia coli (XL-blue) grown in LB medium and purified using EndoFree Plasmid Giga Kit (Qiagen). Standard LAL-test with aliquots of plasmids was performed to assay pyrogenicity of each formulation. In all tested samples endotoxin level did not exceed 10 EU per 1 mg of plasmid DNA, which complies to manufacturer\u2019s range and existing Institute regulations for in vitro and animal tests.All plasmids were amplified in 2HPO4, pH\u200a=\u200a7.1) was added by drops to 0.2 ml of 0.3 M CaCl2 mixed with 2 \u03bcg of pDNA. Resulted mixture was added to a 6-well plate followed by gentle swirling. Cells were incubated overnight under standard conditions and medium was replaced with condition medium (1% FBS DMEM). Aliquots of condition medium were collected 48 hours later, mixed with protease inhibitor cocktail and frozen at \u221270\u00b0C.HEK293T cells were grown in complete DMEM. Transfection was performed on a 6-well plate at 80\u201385% confluent in 2 ml of complete DMEM. 0.2 ml of 2xHBS HUVEC were plated on Matrigel-coated 24-well dish in a mixture of 20% FBS HUVEC medium and low-serum conditioned medium from transfected HEK293T cells in 1\u22361 ratio to obtain final FBS concentration of 10% . Negative control cells were treated with a mock mixture of 20% FBS HUVEC medium and conditioned medium from empty pC4W-transfected HEK293T. Positive control wells were treated with 25, 50 or 100 ng/ml of rhVEGF165 or rhHGF added to 10% FBS HUVEC medium. After 16 hours of incubation under standard conditions tube formation was evaluated in microphotographs taken under 100\u00d7 magnification in phase-contrast mode. Tube length and branching points numbers were counted in MetaMorph software in 5 random FOV taken from each plate well and mean values for control substance or tested medium sample were obtained. All samples were tested in duplicate within every experiment and total of 3 serial runs were taken for final evaluation.Skeletal muscle explant culture experiments were performed in accordance with protocol described by Jang and Kim m. tibialis anterior was excised and washed in 1x PBS and then placed to a mortar pre-cooled on liquid nitrogen. After addition of 500 \u03bcl protein extraction buffer , 1 mM EDTA and 1x protease inhibitor cocktail) muscle was disrupted on liquid nitrogen with a pestle; pulverized sample was collected to a tube and then thawed on water bath at 37\u00b0C. Debris was removed by centrifugation and cleared sample was used for Bradford total protein assay and ELISA detection of human growth factors. For total protein assay the samples were diluted up to 20 fold using protein extraction buffer, for ELISA up to 5 times in accordance to manufacturer's protocol.Skeletal muscle samples from animals injected with phHGF, phVEGF or their combination were harvested at day 3, 7 or 14 and used for transgene protein detection by human-specific ELISA . Negative control animals received empty vector injection. Briefly, animal was sacrificed and We used manufacturer's protocol (R&D Systems) for ELISA detection of VEGF165 and HGF in medium samples and tissue homogenates by corresponding Quantikine\u00ae kit.Samples of culture medium were used for SDS-denaturing electrophoresis in a polyacrilamide gel under non-reducing conditions according to standard procedures. Separated proteins were transferred to a PVDF membrane (Millipore) with subsequent staining by monoclonal mouse anti human antibodies against VEGF165 or HGF overnight at 4\u00b0C and with secondary polyclonal HRP-conjugated goat anti mouse IgG antibodies for 1 hour at room temperature. Two-component West Pico chemiluminescent substrate system (Pierce) was used for development of secondary antibodies.a. femoralis with its branches was ligated distal to inguinal ligament and proximal to its popliteal bifurcation. Vessel was excised between upper and lower ligatures and after control of hemostasis skin was closed with 5\u20130 silk sutures. After completion of surgery animals were put until full recovery into a chamber on an ambient heated pad. After surgery one of plasmid formulations was injected to m. tibialis anterior of ischemic limb. Animals were randomized to receive 100 \u03bcg pC4W-hVEGFopt (n\u200a=\u200a10) or 100 \u03bcg pC4W-hHGFopt (n\u200a=\u200a10). Combined transfer group received a mixture of plasmids: 100 \u03bcg pC4W-hHGFopt+100 \u03bcg pC4W-hVEGFopt (n\u200a=\u200a9). Negative controls were injected with 100 \u03bcg of empty pC4W vector (n\u200a=\u200a9). All injected plasmids (including combined transfer) were diluted in 100 \u03bcl of sterile saline throughout all study groups. Injection was followed by transcutaneous electric pulses as previously described by Schertzer and Lynch Mice were narcotized by intraperitoneal injection of 2.5% avertin solution calculated by body mass. Unilateral induction of hind limb ischemia was performed as previously described Blood perfusion was assessed using a Moor LDI 2.0 system in isoflurane-narcotized animals at 7-day intervals for 3 weeks. Animals were placed to an induction chamber, narcotized by isoflurane inhalation and then placed on an ambient heating pad for 10 minutes under 1\u20132% isoflurane/oxygen inhalation for stabilization of narcosis depth and blood pressure. Consequent perfusion measurements on plantar surface of animal's feet were made and data variability was analyzed using Moor image review software. Readings were taken until 3 subsequent runs with minimal (<10%) deviation were obtained. To account for variability among measurements, ambient light and temperature fluctuations all raw perfusion units readings were normalized against non-ischemic limb and expressed as relative perfusion per cent.m. tibialis anterior was harvested and frozen in TissueTek medium. Serial frozen sections (7 \u03bcm) were prepared and stained for tissue analysis.At day 21 animals from test groups were sacrificed by lethal isoflurane dose followed by cervical dislocation and ischemic We used hematoxylin-eosin staining for tissue necrosis analysis. Necrotic muscle span is expressed as percent to total section area. Signs of muscle necrosis were loss of fiber morphology, cytoplasm disruption, inflammatory cells infiltration and fibrosis.For immunofluorescent analysis sections were formalin-fixed and then incubated overnight with corresponding primary and \u2013 after wash \u2013 secondary antibodies (2 h) and counterstained with DAPI. Stained sections were mounted in aqeous-based medium for subsequent microscopy.Microphotographs of sections were taken under 200\u00d7 magnification in 5 random FOV per section using Zeiss Axiovert 200 M fluorescent microscope with Axiovision 3.1 software. Manual vessel counts in microphotographs were made by two independent persons in MetaMorph 7.1.0.0 software. Capillary density analysis included manual counts of CD31-positive structures per FOV; arteriolar counts were obtained as number of SMA(+) blood vessels with clearly visible CD31(+) inner layer per FOV. Capillary and arteriole counts per FOV were used to obtain mean values for section, animal or group and were subject to subsequent statistical assessment.2 and 1 mg/ml X-Gal (Sigma-Aldrich). Slides were incubated under 37\u00b0C overnight and then washed in PBS and mounted in aqueous-based medium for further microscopy. Transfection efficacy was expressed as ratio of \u03b2-galactosidase positive muscle fibers/total fibers in section.Frozen section of muscle were formalin-fixed and stained for \u03b2-galactosidase activity by incubating them for 24 hours in warm PBS-based solution containing 5 mM potassium ferricyanide crystalline; 5 mM potassium ferricyanide trihydrate; 2 mM MgClData is expressed as mean \u00b1 SD (SEM in some exceptions). Statistical significance of difference between 2 groups was determined using a Student's t-test or Mann-Whitney rank sum U-test depending on sample distribution profile analyzed by Shapiro-Wilk test. Multiple groups were compared using appropriate statistical method with Bonferroni's correction for critical level of significance. Statsoft Statistica 6.0 was used for analyses of study data.We have constructed a novel mammalian expression vector pC4W. Briefly the expression cassette of pC4W consists of human CMV immediate-early promoter/enhancer, rabbit beta-globin intervening sequence-2, consensus translation initiation sequence, woodchuck hepatitis virus posttranscriptional regulatory element (WPRE), and bovine growth hormone and simian virus 40 early polyadenylation signals.To enhance the expression of VEGF and HGF genes we used codon optimization strategy based on the genetic code redundancy. Coding regions of human VEGF165 and HGF gene were screened for the presence of codons occurring with the lowest frequency in human open reading frames. Using conventional genetic engineering and gene synthesis technologies, rare codons were replaced by the most frequent codons encoding the same amino acids, resulting in preservation of the amino-acid sequence of each protein. Furthermore, two signals of mRNA polyadenylation AATAAA and two mRNA destabilizing signals ATTTA in vitro we transfected them to HEK293T cells. Aliquots of condition medium collected after transfection were assayed for human VEGF165 or HGF contents by Western blotting and ELISA. As shown in ex vivo and animal experiments were carried out using plasmids with the highest protein yield in HEK293T cell culture \u2013 pC4W-hVEGFopt and pC4W-hHGFopt.To assay expression activity of different plasmid constructs To evaluate angiogenic activity of VEGF and HGF produced by HEK293T after transfection with pC4W-hVEGFopt and pC4W-hHGFopt we collected medium containing secreted proteins and transferred it to human umbilical vein endothelial cells (HUVEC) on Matrigel-coated surface. Freshly isolated HUVEC were plated in a mixture of double (20%) FBS HUVEC medium and conditioned medium from transfected HEK293T cells in 1\u22361 ratio. Tube formation after 12\u201314 hours of incubation was evaluated by measuring mean tube length and branching point (BP) number per field of view at 100\u00d7 magnification.As expected, HUVEC showed comparable angiogenic response after treatment with 100 ng/ml of positive control proteins (rhVEGF or rhHGF); for details see m. tibialis anterior and transcutaneous pulses were applied as described To assay effect of low-voltage pulses on gene delivery after single intramuscular injection of plasmid DNA we used pC4W-based \u03b2-galactosidase expressing reporter construct. It was injected to Production of human growth factors in murine muscle after gene transfer was detected by ELISA of tissue samples and medium samples collected from muscle explant cultures. Homogenates were prepared from muscle samples taken at day 3, 7 and 14 post injection and after centrifugation were assayed with a human-specific kit (R&D Systems).Significant amount of transgenic protein was detectable in samples taken at every time-point we used yet declining tendency can be obviously seen in presented data A, B. Nom. tibialis anterior harvested 2 days after injection of plasmid DNA . At days 3 and 7 after explantation medium samples were collected for ELISA of human VEGF or HGF contents. Single VEGF or HGF plasmid injection followed by low-voltage electroporation resulted in increasing over time concentration of human angiogenic factor in medium that contained skeletal muscle explant .pC4W-hHGFopr .mixture of pC4W-hVEGFopt and pC4W-hHGFopr .pC4W empty vector as negative control .Limb perfusion measurements were performed at days 7, 14 and 21 after surgery and plasmid injection as described in Materials and Methods. No significant necrotic changes were observed in most animals throughout the study, yet in some control mice amputations of toes were registered.We found that single injection of 100 \u03bcg pC4W-hVEGFopt (VEGF group) or pC4W-hHGFopt (HGF group) plasmids followed by electroporation rendered significant and comparable angiogenic effect by day 21 showing increase of perfusion compared to the empty vector pC4W injection . RelativAs for combined VEGF and HGF gene transfer we injected 200 \u03bcg of total plasmid DNA versus 100 \u03bcg for single gene transfer, we also assessed the influence of plasmid DNA quantity on angiogenic effect expressed as restoration of perfusion. For that purpose we conducted additional animal test with double dose of pC4W-hVEGFopt plasmid. By day 21 after surgery and plasmid administration mice that received 200 \u03bcg of pC4W-hVEGFopt plasmid showed perfusion rates similar to mice injected with 100 \u03bcg of the same plasmid (data not shown).m. tibialis anterior were prepared for further histological analysis.At day 21 after surgery and plasmid administration animals were sacrificed and frozen cross-sections of m. tibialis anterior. Disrupted tissue span expressed as necrotic tissue area/total area section was significantly higher in empty vector group than in any other experimental group indicating severe ischemic damage to skeletal muscle. Ischemic muscle disruption was drastically reduced in animals treated with VEGF, HGF plasmid or their combination capillaries and smooth muscle actin positive (SMA+) vessels were counted after double immunfluorescent staining to proviTo investigate which signalling pathways are activated by combination of VEGF and HGF we analyzed phosphorylation of several MAPKs (mitogen activated protein kinase) in HUVEC stimulated by recombinant HGF, VEGF or both. VEGFR-2 and c-met downstream cascades may have several cross-talk points and using western blot with antibodies specific against phosphorylated forms of ERK1/2 , p38 kinase or Akt we found that treatment with VEGF and HGF together resulted in greater phosphorylation of ERK1/2. In our study we didn\u2019t observe enhanced activation of Akt and p38 after VEGF+HGF addition and found HGF to be a more potent inductor of Akt and P38 . As for Limb ischemia is a great medico-social burden which along with population ageing that correlates with decrease of body regenerative capacity in vivo applicable vectors is also a great point especially taking into consideration that plasmid vectors are safe and feasible with minimal immunogenicity which makes them an attractive object for implication in combined transfer.Completed and ongoing clinical trials focus on single growth factor administration for induction of angiogenesis. Most of them show moderate to low beneficial effect of angiogenic \u201cmonotherapy\u201d for peripheral artery disease after pDNA injection to impaired tissue. Since the original successful attempt by J. Isner In our study we tested a new delivery tool \u2013 a plasmid vector with high protein output and used a combination of two growth factors \u2013 VEGF165 and HGF to augment angiogenesis in ischemic skeletal muscle.VEGF165 is a well-known growth factor and has become an important object of biological and medical research due to its role in angiogenesis, tumor growth and development. For deeper insights into its biological properties one can be advised to pay attention to a series of excellent reviews by N. Ferrara et al. In vitro application of HGF+VEGF results in a more robust proliferative and chemotactic response than each growth factor alone. More complicated 3D collagen models showed that only combination of VEGF and HGF was capable of inducing signification tubulogenic response and facilitate cell survival, yet neither VEGF nor HGF alone was in vivo studies using rhHGF and rhVEGF in rabbit ischemic limb and corneal angiogenesis assay to support previous findings In previous studies combination of HGF and VEGF165 has been tested in endothelial cell cultures. In first part of our work we tested a novel plasmid vector and evaluated its efficacy compared to conventional pcDNA3 vector system. Novel pC4W vector consists of CMV promoter and a number of additional regulatory elements which are combined to obtain higher protein yield. In our cell culture tests we found higher target protein contents after transfection with pC4W-based plasmids compared to pcDNA3-based ones. We also showed additional expression enhancement obtained by codon optimization of angiogenic factor genes selected for our study. Using more effective vectors combined with a safe enhancing technique (e.g. electrotransfer we applied) may result in more successful angiogenesis induction. It seems that there can be a \u201cthreshold\u201d concentration/gradient of growth factor which is required to induce cell proliferation and migration leading to formation of a vessel or to promote survival of both \u2013 pre-existing and dividing endothelial cells. Thus if transfected tissue generates higher amount of therapeutic protein this can lead to better tissue protection against ischemia preventing necrosis, fibrosis or pathological proliferation. In our work we used explant model and tissue homogenate ELISA to confirm human protein secretion by transfected murine muscle. This allowed us to detect secreted human transgene and to ensure that vector activity is sufficient to provide significant amount of target proteins.in situ protein production and explant cultures are a great tool for long-term detection of transgene due to protein accumulation in condition medium over time.Regarding combined angiogenic factors gene transfer little is known about administration of several plasmids in one \u201cshot\u201d. Our data indicates that in early terms (2\u20133 days after injection) protein production after combined delivery is similar to single gene transfer. However, ELISA tests of muscle homogenate samples show that at approximately 1 week there is a significant difference in production of a \u201ccouple\u201d of factors by transfected muscle compared to single factor delivery. Tissue VEGF and HGF concentration after single gene delivery were higher than after combined plasmid injection. This finding that creates an image of discrepancy at first glance makes increase in perfusion we observed after VEGF+HGF treatment even more intriguing. It can point out a possible case for biological amplification of effect as total \u201ctherapeutic protein\u201d amount is constant yet perfusion is enhanced. Thus, our data can indicate that chosen therapeutic factors in lesser quantities can deliver greater benefit when acting together over a period of time after delivery to ischemic tissue. Regarding later time-points we found that by day 14 both proteins are secreted in comparably low quantities and sole plasmid groups have the same protein yield as combined transfer. Similar data was obtained by Korpisalo et al nd week might be the most appropriate time point for administration of a repeated injection of plasmid DNA to support angiogenic response and factor concentration.We evaluated expression period of pC4W-based vector in explants and tissue samples harvested at different time-points after injection. Protein production was maximum in specimen taken at days 3\u20137 with major decline in samples taken at day 14 reflecting drastic fall of protein production. Plasmid vector activity decrement after 2 weeks since administration to skeletal muscle is attributed to plasmid DNA efflux and degradation resulting in lower copy number and subsequent protein extinction. This \u201cfading period\u201d during the 2In vivo tests were performed in mice with unilateral limb ischemia. VEGF- and HGF-induced limb reperfusion after plasmid injection has been reported and reproduced in many works and has been translated to clinical trials. In our study efficient stimulation of angiogenesis after injection of constructed vectors to ischemic skeletal muscle was shown both by blood flow increase and histological findings which included higher CD31+ and SMA+ vessel counts reflecting dense capillary network formation and arteriogenic properties of both factors. Novel finding in this experiment is increasing (up to 75%) perfusion after injection of VEGF+HGF plasmids mixture. We found perfusion in combined VEGF+HGF group to be the highest compared to single plasmid or control. Laser Doppler data was supported by higher capillary and arteriolar density found in combined transfer group animals compared to sole HGF or VEGF groups.in vitro tests in HUVEC cultures indicate that one of putative pathways responsible for angiogenic effect amplification could the ERK1/2 signaling cascade. In our studies we didn\u2019t find increase in Akt or p38 phosphorylation yet in all cases HGF was a more potent inductor of studied pathways than VEGF165. Thus, we can speculate that one of possible mechanisms for amplified angiogenic response is ERK1/2-mediated increase of endothelium proliferation. Most published data covers several possible mechanisms of interaction including thorough study of signaling cross-talk yet comprehensive explanation is still elusive and will be the subject of further studies.Our Ex vivo experiments showed that gene of interest expression retains for at least 14 days after single administration of pDNA. Further we also showed that angio- and arteriogenesis in skeletal muscle can be successfully induced and augmented by administration of combination of pC4W-based plasmids with optimized cDNAs of VEGF165 and HGF.Thus, to conclude, our data indicates that novel pC4W vector is a highly effective and feasible plasmid for therapeutic gene delivery to skeletal muscle. Summarizing our data we can expect the combination of VEGF and HGF to become one of candidates for future translational research along with other candidate pairs: VEGF+angiopoietin-1, VEGF+FGF e t c.in vivo administration as they do in vitro.By the moment all published observations regarding use of several growth factors gene transfer are limited to animal studies due to safety reasons but after first pilot trials and accumulation of safety and prospective data we can expect this approach to become a \u201cnew level\u201d technique in therapeutic angiogenesis. Translation of this method to clinical practice may also require development of next generation bicistronic vectors suitable for human use to administer two genes in one plasmid but use of two separate vectors for reach gene suggested in our study seems to have a great advantage of equal expression of both therapeutic factors which popular IRES-based bicistronic vectors lack after"} +{"text": "We sought to determine normal gender-specific strain values in healthy subjects using Feature Tracking (FT-MRI).Feature tracking (FT-MRI) is a novel MRI based method to analyze myocardial strain that is fast, simple and has potential for clinical use. Similar to echocardiographic \u201cspeckle tracking\u201d myocardial \u201cfeatures\u201d can be tracked on routine steady state free precession or gradient echo images without the need for additional tagged imaging.Global and regional strains including circumferential , radial (Err) and longitudinal (Ell) strains were obtained with FT-MRI (Diogenes v Tomtec Systems) in 60 carefully screened normal subjects. Strains were derived from three long axis and three short axis views after semi automated tracing of endocardial and epicardial borders. Strains were mapped to a 17 segment AHA model and regional strains in the left anterior descending (LAD), circumflex (LCX) and right coronary (RCA) territories derived.There were 29 men and 31 women with mean age 54(14) yrs. The peak global strains were : Ecc (endo) -24(4)%, Ecc (epi)-16(3)%( difference p <0.0001), Ell : -16(5)% and Err : 15(6)%. There was a significant mid to apical gradient in both Ecc (endo) and Ecc (epi) and Ell , but no significant gradient was present in Err. The septum had the lowest Err (8.6(6)% ), while the lateral wall had the highest Err (23.9(9)%) . Err was highest in the LCX territory. Ecc, and Ell were lowest in the RCA territory. Ecc and Ell were generally higher in women than in men, while global Err was lower in women (fig). There was no significant relationship between the global strains and age, blood pressure, cholesterol levels, smoking history and family history of coronary disease. However the strains were significantly associated with height, weight , left ventricular end diastolic volume and ejection fraction . Global Ecc correlated most closely with global ejection fraction . Figure FT-MRI is a novel MRI based method that provides consistent values for global and regional strains in three dimensions from conventional MRI images. There is significant variation in global and regional strains among men and women. There is significant regional and coronary territory heterogeneity of strain patterns."} +{"text": "Bacillus were recovered from 230 inpatient episodes between April and August 2010. An investigation was undertaken.At the National University Hospital in Singapore, the baseline average number of bacillus cultures/month is eight. An outbreak became evident when 274 clinical isolates of Chart reviews of affected patients and extensive environmental sampling was followed by a review of hospital ventilation systems, cleaning protocols and laundry processes. Response to interventions was monitored via clinical case numbers and environmental sampling over a six month period.B. cereus complex constituted 164 cases (71.3%). Bacteraemia comprised 207 patient episodes (90.0%), of which 124 occurred in immunocompromised patients or those with intravascular devices. Physicians treated the organism in 68 episodes (29.5%). Environmental investigations confirmed heavy air contamination particularly within patient rooms and air conditioned wards. Dense airborne contamination outside the hospital adjacent to large earthworks on a construction site was demonstrated (~600CFU/m3). Towels were heavily contaminated even after laundering (7403\u00b11054 spores/cm2). Amplification of spores occurred in clean linen due to storage conditions (165\u00b184 spores/cm2 pre-storage vs 4437\u00b11228 spores/cm2 post-storage). Interventions focusing on laundry protocols, environmental cleaning and air filtration saw clinical case numbers return to baseline levels within three months.Bacillus may be an under-recognised infection risk in hospitals exposed to construction work. Laundering and environmental cleaning processes that are not sporicidal carry a greater risk. Storage conditions of cleaned linen can amplify Bacillus contamination.Environmental contamination with None declared."} +{"text": "Polerovirus tentatively named Pepper yellow leaf curl virus (PYLCV). PYLCV has a wider host range than Tobacco vein-distorting virus (TVDV) and has a close serological relationship with Cucurbit aphid-borne yellows virus (CABYV) (both poleroviruses). The extracted viral RNA was subjected to SOLiD next-generation sequence analysis and used as a template for reverse transcription synthesis, which was followed by PCR amplification. The ssRNA genome of PYLCV includes 6,028 nucleotides encoding six open reading frames (ORFs), which is typical of the genus Polerovirus. Comparisons of the deduced amino acid sequences of the PYLCV ORFs 2-4 and ORF5, indicate that there are high levels of similarity between these sequences to ORFs 2-4 of TVDV (84-93%) and to ORF5 of CABYV (87%). Both PYLCV and Pepper vein yellowing virus (PeVYV) contain sequences that point to a common ancestral polerovirus. The recombination breakpoint which is located at CABYV ORF3, which encodes the viral coat protein (CP), may explain the CABYV-like sequences found in the genomes of the pepper infecting viruses PYLCV and PeVYV. Two additional regions unique to PYLCV (PY1 and PY2) were identified between nucleotides 4,962 and 5,061 (ORF 5) and between positions 5,866 and 6,028 in the 3' NCR. Sequence analysis of the pepper-infecting PeVYV revealed three unique regions (Pe1-Pe3) with no similarity to other members of the genus Polerovirus. Genomic analyses of PYLCV and PeVYV suggest that the speciation of these viruses occurred through putative recombination event(s) between poleroviruses co-infecting a common host(s), resulting in the emergence of PYLCV, a novel pathogen with a wider host range.We determined the complete sequence and organization of the genome of a putative member of the genus Capsicumannum) is an economically important crop worldwide including Israel, where about 3,000 hectares are grown year round for the local and export markets. Since 1998, a viral disease has been found in pepper crops in the south and east of Israel, causing serious economic damage. The disease has been observed mostly in open fields and greenhouses covered with low mesh nets through which insects can easily pass. Disease incidence is higher in greenhouses in which an integrated pest management (IPM) regime or biological control methods are used. The disease symptoms include shortening of stem internodes, inter-veinal yellowing, upward curling of the leaf blade and small, discolored fruit. The disease-causing agent was tentatively named Pepper yellow leaf curl virus (PYLCV) and with TVDV ORF2-ORF4 . Relatively low levels of shared sequence identity were calculated when these sequences were compared to the ORF0, ORF1 and ORF5 sequences of TVDV . The lowest levels of shared sequence identity were calculated for PYLCV and CABYV ORFs (ORF0-ORF5), with the exception of ORF5, for which there was a relatively high level of shared sequence identity (87%) (Polerovirus in the family Luteoviridae and the two poleroviruses Pepper vein yellows virus [PeVYV (AB5948280)] and Tobacco vein-distorting virus [TVDV (EF529624)].Alignment of the deduced amino acid sequences corresponding to the ORF0s of Pepper (TIF)Click here for additional data file.Figure S2Pepper yellow leaf curl virus [PYLCV (HM439608)], Pepper vein yellows virus [PeVYV (AB5948280)], Pepper yellows virus [PepYV (FN600344)], Tobacco vein-distorting virus [TVDV (EF529624)], Potato leaf curl virus [PLRV (Y07496)] and Cucurbit aphid-borne yellows virus [CABYV (X76931)].Alignment of the deduced amino acid sequences of the ORF4s of the poleroviruses (TIF)Click here for additional data file.Figure S3Pepper yellow leaf curl virus [PYLCV (HM439608)], Pepper vein yellows virus [PeVYV (AB5948280)], Cucurbit aphid-borne yellows virus [CABYV (X76931)], Potato leaf curl virus [PLRV (Y07496)] and Tobacco vein-distorting virus [TVDV (EF529624)].Alignment of the deduced amino acid sequences of the N-termini of the ORF5s (CABYV-like region) of the poleroviruses (TIF)Click here for additional data file.Figure S4Potato leaf curl virus [PLRV (Y07496)], Tobacco vein-distorting virus [TVDV (EF529624)], Cucurbit aphid-borne yellows virus [CABYV (X76931)], Pepper vein yellows virus [PeVYV (AB5948280)], Pepper yellow leaf curl virus [PYLCV (HM439608)]. Each protein-coding sequence was aligned using the MUSCLE program and phylogenetic trees were then constructed for each data set using the PhyML software with 100 bootstrap replicates and PLRV, the type member of the genus Polerovirus.Phylogenetic analysis of selected poleroviruses based on the deduced amino acid sequences of the ORF5 of (TIF)Click here for additional data file."} +{"text": "Leptin is the primary hormone in mammals that regulates adipose stores. Arctic adapted cetaceans maintain enormous adipose depots, suggesting possible modifications of leptin or receptor function. Determining expression of these genes is the first step to understanding the extreme physiology of these animals, and the uniqueness of these animals presents special challenges in estimating and comparing expression levels of mRNA transcripts. Here, we compare expression of two model genes, leptin and leptin-receptor gene-related product (OB-RGRP), using two quantitative real-time PCR (qPCR) methods: \u201crelative\u201d and \u201cabsolute\u201d. To assess the expression of leptin and OB-RGRP in cetacean tissues, we first examined how relative expression of those genes might differ when normalized to four common endogenous control genes. We performed relative expression qPCR assays measuring the amplification of these two model target genes relative to amplification of 18S ribosomal RNA (18S), ubiquitously expressed transcript (Uxt), ribosomal protein 9 (Rs9) and ribosomal protein 15 (Rs15) endogenous controls. Results demonstrated significant differences in the expression of both genes when different control genes were employed; emphasizing a limitation of relative qPCR assays, especially in studies where differences in physiology and/or a lack of knowledge regarding levels and patterns of expression of common control genes may possibly affect data interpretation. To validate the absolute quantitative qPCR methods, we evaluated the effects of plasmid structure, the purity of the plasmid standard preparation and the influence of type of qPCR \u201cbackground\u201d material on qPCR amplification efficiencies and copy number determination of both model genes, in multiple tissues from one male bowhead whale. Results indicate that linear plasmids are more reliable than circular plasmid standards, no significant differences in copy number estimation based upon background material used, and that the use of ethanol precipitated, linearized plasmid preparation produce the most reliable results. Both species produce a thick epidermis in addition to blubber and adult bowheads in particular have blubber that may reach up to 50 cm in depth making those depots the largest known in any mammal The majority of leptin studies have focused on the role of the protein in lipid metabolism and energy homeostasis in model mammals (mice and rats) and humans. Leptin is pleiotrophic in its effects and its effect on obesity are well documented in rodent models. Rodents deficient in either leptin Quantitative real-time PCR (qPCR) methodology was developed to allow for highly sensitive and reproducible measurements of relative abundance of mRNA transcripts from a wide variety of species and samples; providing an alternative to more time-consuming traditional methods of gene expression such as RNase protection assays, Northern blots or competitive quantitative PCRs et al., recently demonstrated significant overestimations in copy number for the gene pcna in several species of microalgae when whole circular plasmid standards were used to compare copy numbers estimations from linearized versions of the same plasmid standards A second, less common, method for analysis of target gene expression is \u2018absolute\u2019 quantitative real-time PCR, which shares the highly sensitive and reproducible methods of detection commonly found in relative qPCR assays but differs by not employing the use of an endogenous internal control gene. Absolute qPCR assays allow for quantification of target template copy numbers based on the construction of a standard curve from PCR amplified products, cDNA/DNA, oligonucleotides or plasmid DNA sources \u0394\u0394CT method described by Bustin To assess the expression of leptin and OB-RGRP in cetacean tissues, we first examined how relative expression of those genes might differ when normalized to four common endogenous control genes. We performed relative expression qPCR assays measuring the amplification of two target genes (leptin and OB-RGRP) relative to amplification of 18S ribosomal RNA, ubiquitously expressed transcript (Uxt), ribosomal protein 9 (Rs9) and ribosomal protein 15 (Rs15) endogenous controls. Relative expressions of both target genes were then analyzed following the 2To validate the absolute quantitative qPCR methods used to quantify cetacean leptin and OB-RGRP gene expression we evaluated the effects of plasmid structure (circular and linearized plasmid), the purity of the plasmid standard preparation (precipitated versus un-precipitated linearized standards) and the influence of type of qPCR \u201cbackground\u201d material (RNA or cDNA) on qPCR amplification efficiencies and copy number determination of both model genes in various tissues from one male bowhead whale (09B9). qPCR amplification efficiencies and any significant differences observed in threshold cycle (Ct) values and copy numbers were determined and compared for each test and each gene. Findings from this study strongly support previous findings that linear plasmids are more reliable than circular plasmid standards. In addition we demonstrate that no significant differences were seen in copy number estimation based upon background material used (RNA or cDNA) and that use of ethanol precipitated, linearized plasmid preparations in qPCR assays provides the most reliable results.Balaena mysticetus) and beluga (Delphinapterus leucas) whales by J.G.M. Thewissen (permit: NOAA-NMFS 814\u20131899). The collection permit follows the provisions of the Marine Mammal Protection Act of 1972 as amended (MMPA: 16 U.S.C 1361 et seq) as well as the Endangered Species Act of 1973 as emended (ESA 16 U.S.C 1531 et seq). Post-mortem samples of, liver, whole heart muscle, and testes were taken from one male bowhead (2009B9) approximately 5\u201310 years of age. Post-mortem samples of liver, whole heart tissue and reproductive tissue were taken from a three female bowheads approximately 7\u201312 years for the former and greater than 20 years of age for the later two, an additional male bowhead (10B16) approximately 2 years of age and beluga whales 2009LDL26 (9 years of age), 2010LDL17 (15 years of age) and 2010LDL5 (13 years of age). Age estimations for bowhead whales were done through examination of baleen length Tissue samples were acquired for bowhead using TRI-Reagent\u00ae (Ambion) manufacturer recommended protocols for high fat content samples and quantified via a Nanodrop\u00ae spectrophotometer ND-1000 (Nanodrop). RNAs were PCR analyzed for DNA contamination using designed primers LEPCDNAF and LEPCDNAR that spaEndogenous internal control genes 18S, ubiquitously expressed transcript (Uxt), ribosomal protein 9 (Rs9) and ribosomal protein 15 (Rs15) were amplified from cetacean cDNA using designed primers (Sigma) based on previously published Genbank sequences . PCR ampLeptin and OB-RGPR were amplified from cetacean cDNA using designed primers LEP92F and LEP325R and GRP1F and GRP750R, respectively . PCR ampAll PCR results were evaluated with gel electrophoresis with subsequent ethidium bromide staining. Positive results for each gene were cloned into pCR\u00ae4-TOPO\u00ae vector using a TOPO TA Cloning\u00ae kit (Invitrogen\u2122) following manufacturer recommended protocols and plated on X-Gal-treated LB/AMP plates. Colonies were PCR checked for correct insertion and plasmid preparations completed using QIAprep\u00ae Spin Miniprep Kit (Invitrogen\u2122). Plasmid preparations were quantified via Nanodrop\u00ae spectrophotometer as previously described and sequenced using BigDye\u00ae Terminator v3.1 Cycle Sequencing kit and an ABI 3130xl genetic analyzer to confirm sequence identities prior to real-time PCR assays.\u0394\u0394CT method as described by Bustin Relative qPCR assays were performed with iTaq\u2122 SYBR\u00ae green Supermix with ROX (Biorad). Assays utilized designed leptin-specific qPCR primers Lep258F and Lep325R and OB-RGRP-specific real-time primers GRP620F and GRP750R and endogenous control gene primers for 18S, Rs9, Rs15 and Uxt described previously . 20 \u00b5l rPst1 enzyme (Promega) and manufacturer recommended protocols. Completion of digestion was visualized through gel electrophoresis with ethidium bromide staining as previously described against non-digested plasmids prior to final quantification via Nanodrop\u00ae spectrophotometer. qPCR assays to determine the effect of linear versus circular plasmid structure on copy number estimation and amplification efficiencies were then conducted as described below.Plasmid preparations for vectors containing confirmed inserts for cetacean leptin and OB-RGRP were first analyzed to examine plasmid structure (circular and linearized plasmid). Bacterial plasmids containing either leptin or OB-RGRP were either retained in circular form or linearized through restriction enzyme digestion using The second validation test conducted assessed the importance of precipitating the plasmid standards prior to qPCR analysis. Digested plasmids (digestion described above) were then either used directly in qPCR analysis (for un-precipitated assays) or ethanol precipitated prior to quantification via Nanodrop\u00ae spectrophotometer.Pst1 linearized plasmid standards for leptin and OB-RGRP to examine the effects of background material (either yeast RNA or cDNA) on amplification efficiencies and copy number estimations. Here, the linearized plasmid standards (described above) included either whole yeast RNA or cDNA as background in qPCR assays described below.The third validation test was performed using http://www.uri.edu/research/gsc/resources/cndna.html). For all qPCR assays described below for both genes, the bacterial plasmid standards spanned the full range of copy numbers found in cetacean tissues.Copy number of all bacterial plasmids containing confirmed cetacean leptin and OB-RGRP inserts used in validation test were determined using the following formula: number\u200a=\u200a(ng * number/mole)/(bp * ng/g * g/mole of bp) , designed leptin-specific qPCR primers Lep258F and Lep325R and OB-RGRP-specific real-time primers GRP620F and GRP750R. As before, 20 \u00b5l reactions for all standards were run in triplicate with no template controls and primer controls on an ABI 7300 (Applied Biosystems) under the following conditions for all leptin plasmid validation tests: 60\u00b0C for 2 minutes, 95\u00b0C for 10 minutes, 95\u00b0C for 15 sec for 40 cycles, 60\u00b0C 1 minute. All OB-RGRP qPCR validation assays were run under the following conditions: 55\u00b0C for 2 minutes, 95\u00b0C for 10 minutes, 95\u00b0C for 15 sec for 40 cycles, 60\u00b0C 1 minute. Average Ct values and log (copy number) values were used to obtain a linear regression equation allowing for calculation of copy number of leptin and OB-RGRP in cetacean tissues. The efficiency was calculated using the following equation: Efficiency\u200a=\u200a((10\u2227\u20131/slope)\u20131) Endogenous controls 18S, Rs9, Rs15 and Uxt were selected based on previously published mammalian real-time studies which utilized these genes for normalization qPCR assays of bowhead and beluga tissue cDNAs was performed with iTaq\u2122 SYBR\u00ae green Supermix with ROX (Biorad),leptin, OB-RGRP or endogenous control primers previously described under co10 starting copy number for each standard. Amplification efficiencies for all assays were calculated manually using the following equation: efficiency\u200a=\u200a10(\u22121/slope)\u20131 Threshold cycle values were calculated using Sequence Detection Software version 1.4 (Applied Biosystems) in conjunction with the ABI 7300 (Applied Biosystems). Standard curves for all qPCR assays for all genes and validation tests conducted were produced via linear regressions using threshold cycle (Ct) values and logThe relative expression of two model genes, leptin and OB-RGRP, were examined using four common endogenous control genes, 18S ribosomal RNA, ubiquitously expressed transcript (Uxt), ribosomal protein 9 (Rs9) and ribosomal protein 15 (Rs15). Three tissues from five individual bowhead whales were examined to assess if relative expression values of these model genes varied depending on the control gene utilized. Differences were in fact observed confirming normalization with different controls results in differences in expression .2 values were between 0.9789\u20130.997 for all trials). Threshold cycle values for leptin circular plasmid standards ranged from 20.228 to 29.541 and linearized leptin plasmid standards for leptin ranged from 18.166 to 27.756 indicating no significant difference in threshold cycle values for the two plasmid structures (p\u200a=\u200a0.128). Amplification efficiencies between circular and linearized leptin plasmids were also identical at 97% efficiency. The threshold cycle values for OB-RGRP circular plasmid standards ranged from 22.387 to 29.091 and threshold cycle values for linearized OB-RGRP plasmid standards ranged from 21.136 to 28.724. Again, no significant differences were seen in threshold cycle values when the two plasmid structures were compared (p\u200a=\u200a0.557) and amplification efficiencies were nearly identical at 97.5% for circular OB-GRP plasmid and 97.7% for the linearized plasmid . For OB-RGRP, threshold values for non-precipitated, linearized plasmid standards ranged from 22.295 to 34.067 while precipitated, linearized plasmid threshold values ranged from 19.876 to 31.435. As with leptin, the difference in threshold values between the two plasmid preparations yielded no significant differences (p\u200a=\u200a0.438) . Assays Leptin threshold cycle values using RNA as background ranged from 18.169 to 27.787 and threshold cycles for trials using cDNA as background ranged from 18.166 to 27.756; demonstrating no significant difference in values based on background material selected (p\u200a=\u200a0.998). For OB-RGRP, threshold cycle values ranged from 17.722 to 28.049 using RNA as background and 17.812 to 28.091 using cDNA as background with no significant difference seen (p\u200a=\u200a0.987) . No signBalaena mysticetus) and beluga (Delphinapterus leucas) whale whale . StandarWe applied absolute quantification methods described above to establish standard curves specific to each of the four endogenous control genes used in the relative expression assay . Threshold values ranged from 11.672 to 20.152 for 18S, 11.455 to 20.153 for Rs9, 11.288 to 19.194 for Rs15 and 11.431 to 19.078 for Uxt. No significant difference was seen in the threshold cycle values for the endogenous control genes (p\u200a=\u200a0.990) . SignifiBalaena mysticetus) and beluga (Delphinapterus leucas) whale. Leptin was of particular interest in these species since it is primarily synthesized by adipose cells and these species build and maintain large subcutaneous fat reserves (blubber) throughout their lifetime and the two different species.Examination of physiological systems in marine mammals is complicated by both the life histories of the study animals and access to viable samples. In recent years, relative qPCR assays have been used in marine mammal studies to probe gene expression and provide information regarding ecotype, immunological assessments and physiological response to both metal sensitivity and various pollutants assumed to demonstrate no difference is expression regardless of age, sex or treatment Balaena mysticetus). We normalized amplification of these two model target genes with that of four different endogenous control genes employed previously in relative qPCR studies of other mammalian taxa: 18S ribosomal RNA, ubiquitously expressed transcript (Uxt), ribosomal protein 9 (Rs9) and ribosomal protein 15 (Rs15). Multiple tissues from five individual whales were assayed and results demonstrated significant differences in expression for both genes between both different tissues and different individuals when normalized using the different endogenous controls to determine if this method could also detect significant differences in copy number in the four genes. Whereas no significant differences were observed in the threshold cycle values for the endogenous controls (p\u200a=\u200a0.990), significant differences in copy number were observed among tissues and individuals in the three tissues surveyed. There was also an approximately 10 fold difference seen in the number of copies expressed for 18S compared to Rs9, Rs15 and Uxt expression levels . Of the \u201cAbsolute\u201d qPCR is a second method of analyzing target gene expression. This method forgoes relative expression analysis in favor of measuring actual copy number values via the construction of a standard curve from a variety of different sources pcna gene in various algal species and determined that there was a significant difference in copy number depending on the structure of the plasmid used to construct the standard curve, whether it was circular or enzymatically linearized A recent study examined gene copy number variation of the Validation tests were also conducted to examine the possible effects of plasmid standard purity (precipitated versus un-precipitated linearized standards) on copy number determination and amplification efficiency of linearized bacterial plasmids of cetacean leptin and OB-RGRP. As with the plasmid structure tests, regression curves for both circular and linearized plasmid showed parallel slopes and demonstrated no significant difference in amplification efficiencies with p values of 0.801 (leptin) and 0.438 (OB-RGRP). Results demonstrated a similar pattern of expression in both genes when both the un-precipitated and precipitated standards were used, but significant overestimations of copy number were detected in several tissues for both genes in the cetacean tissues when un-in vivo. To determine the best method for achieving these conditions, validation tests were conducted using both total RNA and total cDNA as \u201cbackground\u201d material to determine what effect, if any, this change had on qPCR amplification efficiencies and copy number determination of leptin and OB-RGRP from cetacean tissues. It is worth noting that the source of this \u201cbackground\u201d material must be carefully selected to insure the absence of any target gene from background source to avoid any co-occurring amplification. In our assays, we employed either total RNA or total cDNA (both extracted from yeast tRNAs) which lack coding sequences for either leptin or mammalian OB-RGRP. Regression curves for both genes yielded nearly identical slopes and p values of 0.998 (leptin) and 0.987 (OB-RGRP) indicative that the choice of RNA or cDNA as background made no significant changes to amplification efficiencies in either gene and a related protein (OB-RGRP) in various tissues of two species of arctic adapted cetacean. Data acquired using only relative qPCR methods provide us only with the former. Data reveals that both species of cetacean produce leptin and that, like other mammals, they produce more leptin in their adipose tissues compared to other tissues (unpublished data). The unique size and extent of cetacean adipose depots, however, makes the question of Our data suggests that individual and sex-specific differences may occur in the expression of the two model genes and that expression levels vary (per 50 ng starting RNA) for the two genes. Furthermore, data regarding the pattern and level of expression for the four endogenous controls demonstrates the range of variation and fold differences seen in between these genes. The absolute quantification methodology described and validated here provides a method for determination of mRNA copy number for genes of interest in any physiological pathway. This technique should prove especially useful in cetacean studies: allowing for examination of a plethora of physiologically relevant genes using only small amounts of starting materials from access to recently dead specimens or non-invasive biopsies or sera samples.in vivo and our data demonstrates that either RNA or cDNA templates are suitable for this purpose.With respect to alternative qPCR methodologies, our results suggest that with the correct precautions the use of absolute quantification can provide both more accurate expression data in addition to those data being more useful in comparative studies. We have provided validation tests to demonstrate this \u201cabsolute\u201d qPCR method reported is valid and provides highly sensitive and reproducible results. We examined the effects of several different variables on the amplification efficiencies and copy number determination of two model genes, leptin and OB-RGRP in various tissues from a bowhead whale in order to determine the best method for conducting absolute quantification qPCR analyses. Based on our results, the best means of standard curve preparation for use in absolute qPCR assays employs the use of a precipitated, enzymatically linearized plasmid containing sequence of a target gene of interest. Additionally, standard curve development and qPCR assays should ideally include the use of a \u201cbackground\u201d material to mimic natural conditions normally found Figure S1Standard curves for linearization tests of leptin and OB-RGRP. Linear regression lines illustrate differences in threshold cycles (Ct) between linear and circular plasmid standards for both leptin and OB-RGRP. Amplification efficiencies calculated using Efficiency\u200a=\u200a ((10\u2227\u20131/slope)\u20131) (TIF)Click here for additional data file.Figure S2Standard curves for plasmid precipitation tests of leptin and OB-RGRP. Linear regression lines illustrate differences in threshold cycles (Ct) between linear and circular plasmid standards for both leptin and OB-RGRP. Amplification efficiencies calculated using Efficiency\u200a=\u200a ((10\u2227\u20131/slope)\u20131) (TIF)Click here for additional data file.Figure S3Standard curves for qPCR background material tests of leptin and OB-RGRP. Linear regression lines illustrate no significant differences in threshold cycles (Ct) between yeast RNA and yeast cDNA background for both leptin and OB-RGRP. Amplification efficiencies calculated using Efficiency\u200a=\u200a ((10\u2227\u20131/slope)\u20131) (TIF)Click here for additional data file.Figure S4Standard curves for qPCR analysis of 18S (A), Uxt (B), Rs9 (C) and Rs15 (D). All linear regressions showed R2 values ranging from 0.9071-0.9915 and amplification efficiencies were calculated using Efficiency\u200a=\u200a ((10\u2227\u20131/slope)\u20131) (TIF)Click here for additional data file."} +{"text": "Hand hygiene compliance is poor among nurses. Hand hygiene compliance is influenced by education. Student nurses\u2019 compliance in clinical practice and perceptions towards hand hygiene have seldom been described.We developed a questionnaire containing demographic data items (i.e. gender and year of education) and items to measure self-reported compliance , attitude towards hand hygiene (7 items), perceived social influence (7 items) and observed environmental conditions during clinical practice (5 items).The questionnaire was filled out by 181 student nurses of the Faculty of Health Care Vesalius, University College Ghent. Students reported a compliance of 50% or below in the following situations: \u2018before and after direct patient contact\u2019 (28%), \u2018after glove removal\u2019 (29%), \u2018after touching patient surroundings\u2019 (65%). Students reported a positive attitude which remained at the same level during the three years. Social influence increased although not significantly . The presence of an observer or mentor during clinical practice was reported to increase social pressure. In general, students were satisfied with the environmental conditions (97%) and with the amount of attention the wards spent on hand hygiene (86%). However, students reported insufficient disinfectant on the carts (21%) and in the patient\u2019s rooms (31%).Undergraduate student nurses reported poor hand hygiene compliance in several specific situations. The students\u2019 attitude towards hand hygiene was positive. Being observed during clinical practice increases social pressure. Some shortcomings in relation to the ward environment were identified.None declared."} +{"text": "Left main stem (LMS) disease is found in approximately 5% of patients with stable angina and in approximately 7% of patients presenting with an acute myocardial infarction. Accurate assessment of the degree of left main stem stenosis has important prognostic and therapeutic implications. Clinically, angiographic LMS stenosis of 50% or more is considered significant. However, it is not known how accurately myocardial perfusion imaging detects LMS disease at this severity threshold.1).1. To measure myocardial blood flow by CMR in patients with LMS stenosis of more than 50% on quantitative angiography in the CEMARC study and blood flow reserve between territories supplied by the LMS and remote territories.2. Following motion correction a circular ROI was selected in the left ventricle to measure the arterial input function. MBF maps were created by model-free analysis; myocardial ROIs were drawn on these maps, one in the LMS territory (ROI1: LAD+LCX) and one in a remote region (ROI2:RCA). MBF for these ROIs was calculated using the Fermi model3. Statistical calculations were performed using SPSS.Nine patients from the CEMARC study who were found to have significant LMS disease on quantitative coronary angiography underwent perfusion-CMR on a Philips 1.5 T Intera system. Myocaridal perfusion imaging was performed every heartbeat during the first pass of 0.05 mmol/kg gadolinium chelate using a T1-weighted fast (spoiled) GE sequence. Stress perfusion imaging was performed using intravenous adenosine infused for 4 minutes (140mcg/kg/min). Perfusion-CMR data were post-processed off-line using the software PMIOf the 9 datasets analysed, the results revealed significant differences p<0.001) in myocardial perfusion seen in LMS diseased territories (ROI1) compared to normal segments (ROI2), Figure in myocaThis study demonstrates reduced myocardial blood flow reserve in patients with LMS stenosis of 50% or more, although reductions are subtle."} +{"text": "The study was carried out to evaluate the antioxidant status in male rats following the dietary consumption of red palm oil.Male wistar rats were randomly divided into three groups. Group 1 (n=5) received no supplementation and served as the control while group 2 (n=6) and 3 (n=6) received 2ml and 4ml red palm oil (RPO), respectively. Plasma total polyphenols, plasma antioxidant capacity [i.e. oxygen radical absorbance capacity (ORAC)], ferric reducing antioxidant power (FRAP) as well as levels of antioxidant enzymes were determined using established techniques.There were no significant differences (p<0.05) in total polyphenols, ORAC, and FRAP in palm oil fed groups when compared with the control group. Catalase levels significantly increased (p<0.05) at both 2ml and 4ml RPO in the liver and erythrocyte. There was no significant difference in the liver GPx levels in palm oil fed groups while erythrocyte GPx level significantly increased at 4ml RPO when compared with the control group. Red palm oil did not significantly increase (p<0.05) liver and erythrocyte SOD levels in all the groups when compared with the control group.Red palm oil did not significantly increase the total antioxidant capacity in the plasma. However, RPO significantly increased the levels of liver and erythrocyte catalase as well as erythrocyte glutathione peroxidase level and hence, its dietary consumption could help to boost antioxidant status in the body and thus promote good health."} +{"text": "Sebaceous neoplasms of the skin (SN) are described in the Muir Torre variant of Lynch syndrome (LS). Guidelines recommend evaluating individuals diagnosed with sebaceous adenomas or sebaceous carcinomas for LS with immunohistochemistry (IHC) for mismatch repair (MMR) proteins and/or microsatellite instability analysis (MSI). The assumption has been that SNs with defective MMR are related to LS.To describe outcomes of genetic testing for LS among individuals with SN.24 individuals with a personal history of SN underwent a genetic evaluation for Lynch syndrome (LS) at Dana-Farber Cancer Institute. 10 had family histories which met Amsterdam criteria, 8 had a personal history of another LS-associated malignancy, 23 had family history of one or more LS-associated cancers, and 1 had no other personal or family history of cancer.MLH1 and 9 MSH2) and each of these either met Amsterdam criteria or had a personal history of another LS-related cancer.11/24 (46%) probands had pathogenic MMR gene mutations . One of the probands whose SN showed absence of MSH2 and MSH6 had a family history which met Amsterdam criteria (PREMM score=33%) and the rest had PREMM model scores <5%.Although prior reports suggest that MSI/IHC can be useful in screening patients with SNs for LS, we found many of these tumors demonstrate features of abnormal MMR even when family history is not suggestive of LS and genetic testing did not reveal MMR mutation. Further study is needed to determine whether other somatic mechanisms may produce the MMR deficient phenotypes seen in many SNs."} +{"text": "During the 2010/2011 winter the H1N1 influenza pandemic placed increased demand on critical care services, prompting our department to devise a modified triage tool for the ICU to be implemented at a time of exceptional bed crisis [2 <90% on FiO2 >85%, respiratory acidosis pH \u22647.2, respiratory failure or airway compromise, systolic pressure <90 mmHg, SOFA score \u22657) or refusal criteria were recorded with outcome data.We retrospectively examined patient notes for all admissions to our adult ICU during December 2010 and January 2011. Patient admission criteria fulfilled at least one admission and no refusal criteria. Two patients (7%) had documented refusal criteria. The first of these had a severe life-limiting condition, staying 29 days in the ICU and a further 65 days in hospital. The second was admitted post non-VF arrest, dying after 2 days in the ICU. Three patients (11%) met no admission criteria. These patients stayed between 4 and 6 days in critical care with total hospital stays of 18 to 98 days, one requiring 30 days of rehabilitation.The proposed admission criteria concurred with clinical decision-making in 81% of admissions. The patients that met refusal criteria required either prolonged hospital stay or had short survival times and may not represent optimal utilization of critical care facilities during a time of increased demand. Those patients not meeting the admission criteria had short critical care stays illustrating that rigid admission requirements may exclude patients who could benefit from critical care. A standardized set of admission criteria may supplement decision-making during times of increased critical care demand and strengthen documentation of those decisions. However, no set of criteria can replace clinical judgement in critical care admission."} +{"text": "Subsequent cytogenetic and PCR analyses demonstrated that homozygosity of a ~500 kb chromosomal segment translocated from BTA6 to BTA29 is the underlying genetic mechanism responsible for gonadal hypoplasia. The duplicated segment includes the KIT gene that is known to regulate the migration of germ cells and precursors of melanocytes. This duplication is also one of the two translocations associated with colour sidedness in various cattle breeds.Impaired migration of primordial germ cells during embryonic development causes hereditary gonadal hypoplasia in both sexes of Northern Finncattle and Swedish Mountain cattle. The affected gonads exhibit a lack of or, in rare cases, a reduced number of germ cells. Most affected animals present left-sided gonadal hypoplasia. However, right-sided and bilateral cases are also found. This type of gonadal hypoplasia prevails in animals with white coat colour. Previous studies indicated that gonadal hypoplasia is inherited in an autosomal recessive fashion with incomplete penetrance. In order to identify genetic regions underlying gonadal hypoplasia, a genome-wide association study (GWAS) and a copy number variation (CNV) analysis were performed with 94 animals, including 21 affected animals, using bovine 777,962 SNP arrays. The GWAS and CNV results revealed two significantly associated regions on bovine chromosomes (BTA) 29 and 6, respectively (P=2.19 x 10 Gonadal hypoplasia is characterised by aberrantly small and underdeveloped gonads. Fertility is generally disturbed when both gonads are hypoplastic. Different types of gonadal hypoplasia have been reported in several mammalian species, including cats, dogs , sheep , horses Hereditary gonadal hypoplasia is a frequent disorder in two Scandinavian cattle breeds, Northern Finncattle and Swedish Mountain cattle . The defect emerged in the early 20th century after pure-breeding of Swedish Mountain cattle was implemented ,7. A fewThe incidence of gonadal hypoplasia in the mentioned breeds is equal in both sexes and comprehensive breeding experiments suggest a recessive mode of inheritance with incomplete penetrance . PenetraGonadal hypoplasia is unique in Northern Finncattle and Swedish Mountain cattle because it is mainly manifested on the left side. The proportions of left-, double- and right-sided gonadal hypoplasia are 82%, 15% and 3%, respectively . AlthougThe severity of gonadal hypoplasia varies considerably from total to partiGonadal hypoplasia of Northern Finncattle and Swedish Mountain cattle is a congenital defect and SettCs [29 allele in BTA29 resulted from a duplication and translocation event of a 492 kb segment of BTA6 including the KIT gene. The Cs6 allele residing on BTA6 is a result of a subsequent duplication and translocation event that moved back the segment comprising fused sequences of the BTA29 and BTA6 to the KIT locus in BTA6. It is expected that in both cases the dysregulation of the KIT gene leads to the colour sidedness [Gonadal hypoplasia occurs only in animals that are 60-100% white coloured [6]. TheCs ,14. ReceCs showed tidedness .KIT gene encodes a type III receptor protein of the tyrosine kinase family. Several studies have shown that KIT protein is crucial for survival, proliferation and migration of melanocyte precursors and primordial germ cells (PGC) during embryogenesis and and15] aFive case samples, two control samples with a duplication of the BTA6 segment, two control samples with both studied duplications and three control samples without the duplications were subjected to sequencing. A 10-20 ng of purified PCR product was mixed with 0.5 \u00b5l BigDye\u00ae Terminator v3.1 (Life Technologies) and 0.5 \u00b5l of the forward or reverse PCR primer (2.5 pmol). The sequencing reaction was performed under the following conditions: initial denaturing at 96 \u00b0C for 10 sec, followed by 35 cycles, including 10 sec in 96 \u00b0C, 5 sec in 50 \u00b0C, and 4 min in 60 \u00b0C. The gel filtration of the sequencing reaction was applied using the MultiScreen filtration plate and Sephadex G-50 Fine (Sigma) followed by capillary electrophoresis carried out on 3130xl Genetic Analyzer (Life Technologies). Base calling, sequence alignment and polymorphism detection were made using the Phred/Phrap/Polyphred software . SequencCAATTTTAAGCATGTGCTGAGG and ACAGCCTCTGGTCTGTCTGG to validate the genotype calls for the SNP on BTA23 at 52,435,290 bp.Seven cases and ten controls were sequenced (see above) with primers Figure S1Examples of the common colour pattern in Northern Finncattle.Most commonly, Northern Finncattle is almost white with black or brown in ears and muzzle. The flanks and legs can also be partly coloured or spotted.(TIF)Click here for additional data file.Figure S2Average log R ratio of animals carrying the ectopic BTA29 segment.The average log R ratio was calculated from 15 affected and 44 unaffected animals that carry the duplicated segment of BTA29. The 5-SNP-sliding window log R ratio is presented for 563 SNPs.(PNG)Click here for additional data file.Figure S3KIT containing segment to chromosome 29.Localisation of a translocation of a Animals carrying two and four copies of the BTA6 segment were compared using Fisher exact tests. The red dots represent significantly associated SNPs (P < 7.71 x 10-8).(EPS)Click here for additional data file.Figure S4Localisation of a translocation of a BTA29 segment to chromosome 6.Animals with and without the presence of a BTA29 CNV were compared using Fisher exact tests. The red dots represent significantly associated SNPs (P < 7.71 x 10-8).(EPS)Click here for additional data file.Figure S5FISH studies.Three Northern Finncattle (NFC) animals with different combinations of the Cs29 allele and one animal of the Western Finncattle (WFC) and Eastern Finncattle (EFC) were analysed by FISH with two BAC probes. The Cs29 allele is associated with both colour sidedness and gonadal hypoplasia and it corresponds to the red FISH signal or the red bar. The Cs6 allele is associated with colour sidedness and corresponds to the green FISH signal or the green bar. Overlapping red and green signals appear yellow. All animals except the solid brown Western Finncattle had one or several Cs alleles. The animal NFC 164 is affected with gonadal hypoplasia and it is homozygous for the Cs29 allele.(TIF)Click here for additional data file.Figure S629 allele.Association of 647,971 SNPs with the affection status of 39 animals homozygous for the CsAssociation analysis was performed using Fisher exact tests of allelic association for 21 affected and 18 unaffected animals homozygous for the Cs29 allele.(JPG)Click here for additional data file.Table S1The association between the proportion of coat pigmentation and total gonadal hypoplasia in the Swedish Mountain breed females (modified from Settergren ).(DOCX)Click here for additional data file.Table S2Nominal p-values for the SNPs significantly associated to gonadal hypoplasia in BTA29 .P-values were calculated with Fisher exact tests in PLINK to determine allelic association in four different case-control cohorts ( cohorts .(XLSX)Click here for additional data file.Table S3Nominal p-values for the SNPs significantly associated to gonadal hypoplasia in BTA29 (genotypic test).P-values were obtained using a 2df genotypic test implemented in PLINK to determine association in four different case-control cohorts ( cohorts .(XLSX)Click here for additional data file.Table S4Nominal p-values for the SNPs located between 71502659 bp and 71990541 bp in BTA6 .P-values were calculated with Fisher exact tests in PLINK to determine allelic association in four different case-control cohorts ( cohorts .(XLS)Click here for additional data file.Table S5Nominal p-values for the SNPs located between 71502659 bp and 71990541 bp in BTA6 (genotypic test).P-values were obtained using a 2df genotypic test implemented in PLINK to determine association in four different case-control cohorts ( cohorts .(XLS)Click here for additional data file.Table S6Predominantly white coloured animals divided according to affection status and alleles in BTA6 and BTA29.The CNVs were studied with PCR and primers designed by Durkin et al. [6 allele could not be distinguished.n et al. and us. (DOCX)Click here for additional data file."} +{"text": "Acute mesenteric ischemia (AMI) is an abdominal emergency caused by: embolism (40-50%), with Superior Mesenteric Artery (SMA) thrombosis (20-25%), mesenteric venous thrombosis (5%) and non occlusive mesenteric ischemia (20%). The mortality rate is high and ranges from 64 to 93%.We present a case of a 75-year-old patient with acute occlusive mesenteric ischemia that was successfully treated with endovascular intervention.Angiography revealed high-grade stenosis of the proximal tract of the SMA. Immediate option for endovascular therapy was made, and a MARIS self-expandable 6x40 mm stent was positioned. The patient was discharged 2 days after with full recovery from the symptoms.Control angiography showed the correct apposition of the stent and regular flow inside. The patient was symptom free 12 months later.We suggest that the endovascular technique is a valid option in patients with AMI preventing intestinal infarction."} +{"text": "The majority of clinical guidelines recommend lateral wedge shoe insoles for medial knee osteoarthritis (OA), despite limited and equivocal evidence of efficacy. The objective of this study was to assess efficacy of lateral wedge insoles for improving symptoms and slowing structural disease progression compared with control insoles in medial knee OA.A randomised participant- and assessor-blinded controlled trial was used. 200 people aged 50 or more with clinical and radiographic diagnosis of mild-to-moderately severe medial knee OA were recruited. The interventions consisted of full-length 5\u00b0 lateral wedged insoles or flat control insoles worn inside the shoes daily for 12 months. The primary symptomatic outcome was change in overall knee pain (past week) measured on an 11-point numeric rating scale and primary structural outcome was change in medial tibial cartilage volume from magnetic resonance imaging. Secondary clinical outcomes included changes in measures of pain, function, stiffness, and health-related quality of life. Secondary structural outcomes included progression of medial cartilage defects and bone marrow lesions.3 (-15.4 to 14.6)). None of the changes in secondary outcomes demonstrated differences between groups.There were no significant between-group differences for the primary outcomes of change in overall pain (-0.3 points 95% CI (-1.0 to 0.3)) and change in medial tibial cartilage volume (-0.4 mmIn this study, lateral wedge insoles worn for 12 months provided no symptomatic or structural benefits compared to a flat control insole."} +{"text": "Cough is a phenomenon frequently associated with upper airway diseases and as a reflex is modulated by many afferent inputs either from respiratory tussigenic areas, but also by afferent drive from other organs. Modulation of cough by nasal afferent inputs could either facilitate cough response or inhibit it in animal models, depending on the type of trigeminal afferents which are stimulated. In recent study we focused on afferents expressing TRPA1, TRPM8 & TRPV3 channels -- channels known as relevant for airway irritants (TRPA1), menthol and other cooling substances (TRPM8) and thymol (TRPV3). Particularly menthol and thymol are substances which are frequently used in over-the-counter medication for cough and common cold based on empirical approach. Objective evidence regarding the modulation of cough in humans has never been reported. 60 human healthy volunteers participated in the study, and they have been challenged by intranasal drops containing agonists of selected ion channels: isocyanate (AITC) & cinnamaldehyde for TRPA1, (-) menthol and (+) menthol for TRPM8 and thymol for TRPV3 ion channels in randomized order . Nasal symptom score, cough threshold (C2), urge to cough (Cu) and cumulative cough response had been assessed using capsaicin cough challenge tests. Nasal challenges of TRPA1 relevant agonists induced considerable nasal symptoms, significantly enhanced urge to cough (p < 0.05) but modulation of C2 and cumulative cough response did not reach significance level. Both TRPM8 agonists and TRPV3 agonist thymol administered to the nose significantly modulated all parameters including C2 (p<0.05), Cu (p <0.01) and cumulative cough response (p < 0.01) documenting strong anti irritating and antitussive potential of menthol isomers and thyme. Nasal afferent drive modulates cough reflex in human healthy volunteers and this knowledge could have clinical application involved in relieving lower airway symptoms in subjects with upper airway diseases. The role of trigeminal afferents, olfactory nerve endings, smell perception process and other supramedullar influences have to be taken into consideration as relevant enough to modulate cough response in humans."} +{"text": "Genotype imputation, used in genome-wide association studies to expand coverage of single nucleotide polymorphisms (SNPs), has performed poorly in African Americans compared to less admixed populations. Overall, imputation has typically relied on HapMap reference haplotype panels from Africans (YRI), European Americans (CEU), and Asians (CHB/JPT). The 1000 Genomes project offers a wider range of reference populations, such as African Americans (ASW), but their imputation performance has had limited evaluation. Using 595 African Americans genotyped on Illumina\u2019s HumanHap550v3 BeadChip, we compared imputation results from four software programs and three reference panels consisting of different combinations of 1000 Genomes populations (February 2012 release): (1) 3 specifically selected populations ; (2) 8 populations of diverse African (AFR) or European (AFR) descent; and (3) all 14 available populations (ALL). Based on chromosome 22, we calculated three performance metrics: (1) concordance (percentage of masked genotyped SNPs with imputed and true genotype agreement); (2) imputation quality score ; and (3) average r2hat . Across the reference panels, IMPUTE2 and MaCH had the highest concordance (91%\u201393%), but IMPUTE2 had the highest IQS (81%\u201383%) and average r2hat . Imputation quality for most programs was reduced by the addition of more distantly related reference populations, due entirely to the introduction of low frequency SNPs (MAF\u22642%) that are monomorphic in the more closely related panels. While imputation was optimized by using IMPUTE2 with reference to the ALL panel (average r2hat\u200a=\u200a0.86 for SNPs with MAF>2%), use of the ALL panel for African American studies requires careful interpretation of the population specificity and imputation quality of low frequency SNPs. Genotype imputation is often conducted in genome-wide association studies (GWAS) as an efficient approach to expand coverage of single nucleotide polymorphisms (SNPs), enabling meta-analysis of GWAS from different genotyping platforms Genotype imputation in admixed populations has not performed as well, because their genetic diversity is greater than the original reference populations Imputation in African Americans has typically relied on some composition of the YRI, CEU, and CHB+JPT populations from HapMap p-value<0.0001. There remained 541,860 autosomal SNPs (96.5%), of which 8,101 SNPs from chromosome 22 were used for imputation.We obtained genome-wide genotyping data available from Illumina\u2019s iControl database for 830 African Americans genotyped for 561,466 SNPs on the Illumina HumanHap550v3 BeadChip. Data were downloaded on January 19, 2011. Quality control (QC) was implemented on SNPs and subjects using PLINK The subsequent subject-level QC procedures are outlined in The iControlDB subjects were then evaluated for population structure to identify ancestral outliers. We implemented the pairwise population concordance test in PLINK, which is based on the observed proportion of IBS loci pairs, and we identified eight subjects who were significantly different (P<0.0005) from 95% of the rest of the population. The ancestral outliers were confirmed using the STRUCTURE program ftp://ftp.1000genomes.ebi.ac.uk/vol1/ftp/release/20110521/, accessed March 7, 2012). We created three reference haplotype panels based on different combinations of 1000 Genomes populations. The first 1000 Genomes reference panel was created by combining YRI (N\u200a=\u200a88), CEU (N\u200a=\u200a85), and ASW (N\u200a=\u200a61) populations, which were specifically chosen to provide a close match to African American study populations. We previously evaluated several different combinations of HapMap phase III reference populations for our African American study population and found that the combined YRI+CEU+ASW panel had optimal imputation performance We obtained prephased reference haplotypes from the February 2012 release of the 1000 Genomes project and the August 2010 release of 1000 Genomes , as shown in Figures S3, S4, S5.The second 1000 Genomes reference panel, having eight populations of African (AFR) or European (EUR) descent, was chosen as a broader match for African Americans. The combined AFR+EUR panel included YRI, CEU, ASW, as well as Kenyans , Finns , Britons , Spaniards , and Italians . Additional comparisons of the YRI+CEU+ASW and AFR+EUR reference panels were made using HapMap phase III . This \u201ccosmopolitan\u201d approach of using all available reference populations has been advocated by others, particularly for diverse study populations http://www.sph.umich.edu/csg/yli/MaCH-Admix/.As reviewed elsewhere Imputation using IMPUTE2 Imputation using BEAGLE For each imputed SNP, we obtained a genotype dosage value and the most likely discrete genotype, either directly from the output of the imputation software or via conversion of the software output (IMPUTE2). For all imputation procedures, we masked 2% of the genotyped SNPs to allow direct comparisons of their true and imputed genotypes. The imputation procedures were repeated 10 times with 10 different sets of randomly masked SNPs, with one exception due to excessive computational runtime . Indel variants from 1000 Genomes panels were not included, in order to focus on evaluating the performance of SNP genotype imputation.http://www.sph.umich.edu/csg/yli/software.html) was needed to generate a single, common metric to assess imputation quality across the programs.For each imputation scenario based on the different software programs and reference panels, we calculated three imputation performance metrics, which captured different features of imputation accuracy and quality. First, after masking 2% of the genotyped SNPs, we calculated the concordance rate as the percentage of genotype calls for which the true genotype matches the most likely discrete imputed genotype. This concordance rate calculation, based on discrete imputed SNP genotypes, has been used often as a measure of imputation accuracy Given the need for computational efficiency with the large number of imputations (10 repetitions for each imputation procedure), our analyses focused on chromosome 22, as other studies evaluating imputation performance have done The performance results comparing the 12 imputation strategies (four different programs and three different reference panels from the February 2012 release of 1000 Genomes) are shown in When focusing on the 1000 Genomes February 2012 release, the highest concordance rates were obtained using IMPUTE2 and MaCH, and these two programs performed equally well across the reference panels . MinimalUnlike concordance, the IQS results revealed that IMPUTE2 had the highest imputation accuracy when taking MAF into account for the 1000 Genomes (February 2012 release) reference panels . The IQSWith regard to average r2hat, IMPUTE2 outperformed all other programs , but incWe evaluated two practical approaches to minimize the occurrence of low frequency SNPs that are most likely monomorphic in African Americans. First, we used the straightforward approach of imposing an r2hat threshold on the imputed SNP set, which reduced, but did not eliminate, the occurrence of problematic low frequency SNPs. For instance, 32.7% of the SNPs imputed based on the ALL panel were monomorphic in the YRI+CEU+ASW panel; after imposing the r2hat\u22650.3 threshold, 11.0% of the remaining SNPs were monomorphic in the more closely related panel. As an alternative approach, SNPs can be filtered out based on their MAF in reference panel subpopulations before or after imputation. For pre-imputation filtering, IMPUTE2 offers a filtering option (\u201c-filt_rules_1\u201d) that removes reference SNPs that are monomorphic in the panel of interest , hence leaving fewer input SNP genotypes and speeding the imputation procedure. For postimputation filtering, SNPs are imputed using all available reference SNPs as input and then those that are monomorphic in the panel of interest are removed. We applied this postimputation filtering strategy to our imputation results. When considering only the 312,474 SNPs that are polymorphic across the reference panels out of 475,371 total SNPs on chromosome 22, use of the ALL panel resulted in the highest average r2hat, both overall , among other uses. Genetic studies using controls from public sources require stringent QC procedures, as demonstrated by our population structure analyses showing that several of the publicly available African American study subjects were ancestral outliers, as previously reported p-values of true association signals might be attenuated for the low frequency SNPs. Further, inclusion of more distantly related reference panels for imputation in African-derived study populations has been suggested to weaken association signals particularly near loci under strong selective pressure Our study is the first to highlight important considerations when using the cosmopolitan approach with 1000 Genomes to conduct imputation in an admixed study population, particularly an African American population. Imputation quality of all low frequency SNPs was relatively low, and imputation of low frequency, population-specific SNPs was especially prone to imputation error. Because imputation error leads to a loss of statistical power Figure S1Quality control procedures for African Americans genotyped on the Illumina HumanHap550v3 BeadChip from Illumina\u2019s iControlDB. Quality control procedures were conducted using PLINK, unless otherwise stated. At each step, the number of excluded subjects is provided. For each pair or cluster of subjects identified in steps 2 and 3, we retained only the one subject having the highest call rate.(DOC)Click here for additional data file.Figure S2STRUCTURE triangle plot showing estimated ancestral proportions of African American study subjects relative to HapMap populations. African American study subjects from iControlDB (in red) were genotyped on the Illumina HumanHap550 BeadChip version 3. Ancestral proportion estimates were based on 10,000 randomly selected HapMap SNPs in linkage equilibrium. The triangle\u2019s vertices represent West Africans (YRI subjects in blue), European Americans (CEU subjects in yellow), and East Asians (CHB subjects in green), and the triangle\u2019s edges indicate the ancestral proportions. African Americans from HapMap (ASW subjects in black) were also included for admixture comparison. African American study subjects with an African ancestry <60% were excluded from further analysis.(DOC)Click here for additional data file.Figure S3Concordance resulting from four different imputation programs and three different reference panels from either HapMap phase III or 1000 Genomes (August 2010 release). Concordance rates were based on masking 2% of the genotyped SNPs on chromosome 22 and comparing imputed and true genotypes. The number of subjects corresponding to each reference panel is shown in parentheses.(DOC)Click here for additional data file.Figure S4Imputation quality score (IQS) resulting from four different imputation programs and three different reference panels from either HapMap phase III or 1000 Genomes (August 2010 release). IQS results were based on masking 2% of the genotyped SNPs and adjusting the concordance rate chance agreement between imputed and true genotypes. The number of subjects corresponding to each reference panel is shown in parentheses.(DOC)Click here for additional data file.Figure S5Average r2hat values resulting from four different imputation programs and three different reference panels from either HapMap phase III or 1000 Genomes (August 2010 release). r2hat values were averaged across all imputed SNPs on chromosome 22. The number of subjects corresponding to each reference panel is shown in parentheses.(DOC)Click here for additional data file.Figure S6Average r2hat, based on imputation using MaCH, across the minor allele frequency (MAF) spectrum. Imputation was conducted for all SNPs available on the YRI+CEU+ASW , AFR+EUR , or ALL reference panels from 1000 Genomes. Imputed polymorphic SNPs were divided into MAF intervals of 1%, and their average r2hat values were calculated within each interval.(DOC)Click here for additional data file.Figure S7Average r2hat, based on imputation using MaCH-Admix, across the minor allele frequency (MAF) spectrum. Imputation was conducted for all SNPs available on the YRI+CEU+ASW , AFR+EUR , or ALL reference panels from 1000 Genomes. Imputed polymorphic SNPs were divided into MAF intervals of 1%, and their average r2hat values were calculated within each interval.(DOC)Click here for additional data file.Figure S8Average r2hat, based on imputation using BEAGLE, across the minor allele frequency (MAF) spectrum. Imputation was conducted for all SNPs available on the YRI+CEU+ASW , AFR+EUR , or ALL reference panels from 1000 Genomes. Imputed polymorphic SNPs were divided into MAF intervals of 1%, and their average r2hat values were calculated within each interval.(DOC)Click here for additional data file.Figure S9Average r2hat values resulting from four different imputation programs and three different 1000 Genomes (February 2012) reference panels, considering only imputed SNPs that were polymorphic on the YRI+CEU+ASW panel. r2hat values were averaged across the 312,474 relevant imputed SNPs out of 475,371 imputed SNPs on chromosome 22. The number of subjects corresponding to each reference panel is shown in parentheses.(DOC)Click here for additional data file.Figure S10Average r2hat, based on imputation using IMPUTE2, across the minor allele frequency (MAF) spectrum. Imputation was conducted for all SNPs available on the YRI+CEU+ASW , AFR+EUR , or ALL reference panels from 1000 Genomes, but only SNPs present across all reference panels are shown. Imputed polymorphic SNPs were divided into MAF intervals of 1%, and their average r2hat values were calculated within each interval.(DOC)Click here for additional data file.Table S1iControlDB subjects genotyped on the Illumina HumanHap550v3 BeadChip, who were identified as African American but their genetic data indicated less than 60% African ancestry. These 179 subjects were excluded due to ancestral misclassification.(DOC)Click here for additional data file."} +{"text": "The African continent bears the greatest burden of HIV/AIDS in the world. Efforts by scientists to find a vaccine for curing the pandemic have proved futile to date. The prevalence in Uganda stands at 6.4%in Adults and 0.7 % in Children, and about 1.1 million Ugandans are living with HIV/AIDS. The AIDS Support Organization started in 1987. Non disclosure of HIV sero status affects uptake of HIV/AIDS health services, increases stigma and discrimination.A retrospective cohort study was done to review records of patients newly registered between January and December 2007 from Management Information System. We analyzed records of patients who had not previously disclosed their HIV/AIDS Sero status at the time of entry into TASO HIV/AIDS clinic. These patients routinely received counseling services for a period of 36 months to assess their disclosure status.Out of 1413 patients counseled, 117(8.3%) patients had not disclosed their HIV sero status, 27(23%) patients were sexually active. During the first individual counselling sessions, patients were given information on condom use 19%, septrine prophylaxis 18%, sexually transmitted infections 15%, family planning 12%, and antiretroviral therapy 9%, safe water 8%, abstinence 5%, life skills 4%, prevention of mother to child transmission 4%, Tuberculosis 4%, voluntary counselling and testing 2%, and faithfulness 1%. Significant number of these sessions focused on opportunistic infections 39%, disclosure 15%, antiretroviral therapy 14%, drug therapy 9%, STD and HIV prevention 5%, nutrition 4% and welfare 4%, discordance 2%, sexuality and abstinence 3%. Other indirect interventions included HIV prevention sensitization through formation of peer support groups, drama sensitizations, group counseling and health talks during clinics. After 36 months of follow-up, 65-56% of 117 patients had disclosed their HIV sero status. Disclosure of HIV status is statistically associated with the number of counseling sessions (p=0.008). Average number of counseling sessions was 6 sessions. Patients who had not disclose after 36months recorded an average of 3 counselling sessions. Revealing of HIV sero status is statistically associated by sex, more female reveal their HIV Sero status in as short life span compared to males (p=0.002).The number of counseling sessions someone receives is associated with supported disclosure. Female patients reveal their HIV sero status in a shorter time span compared to males. Integration and more frequent provision of counseling services to patients in HIV/AIDS care and treatment enables them to make informed decisions regarding disclosure of their HIV sero status to their family members, sexual partners, friends and others. This has created support systems to clients and therefore reducing further spread of HIV."} +{"text": "Patients with neurofibromatosis type 2 (NF2) develop bilateral cochleovestibular schwannomas (CVSs) that cause binaural deafness in most individuals. Hearing loss occurs in an unpredictable manner and the underlying mechanisms are not known. To gain insight into the pathophysiologic basis for hearing loss in NF2, we performed a prospective cross-sectional study of untreated ears in NF2 patients.One hundred consecutive NF2 patients in a prospective natural history study were included. Clinical and audiometric data were analyzed for treatment na\u00efve ears. In addition to standard MR-imaging sequences, alterations in intralabyrinthine protein content were determined utilizing high resolution FLAIR, the presence of cochlear aperture obstruction was determined by examining 3D T2 sequences, and endolymphatic hydrops was identified on delayed post-contrast FLAIR sequences.3, including 14 ears (88%) with block of the cochlear aperture and elevated protein.Eighty-nine ears harboring 84 untreated CVSs in 56 consecutive NF2 patients (age 30\u00b116 years) were analyzed. Thirty-four (38%) ears had varying degrees of hearing loss. Elevated intralabyrinthine protein was identified in 70 (75%) ears by FLAIR MR-imaging and was strongly associated with the presence of hearing loss . Elevated intralabyrinthine protein was associated with the presence of CVS-associated cochlear aperture obstruction in both normal and hearing loss ears. Elevated intralabyrinthine protein was not identified in ears without CVS (5 ears). While larger tumor size was associated with hearing loss (P\u200a=\u200a0.006), 16 hearing loss ears (47%) harbored CVSs less than 0.5 cmThese findings are consistent with a model in which hearing loss develops as a result of cochlear aperture obstruction and accumulation of intralabyrinthine protein. MRI based identification of elevated intralabyrinthine protein may help identify the ear at-risk for developing hearing loss. NF2 tumor suppressor gene located on the long arm of chromosome 22 Neurofibromatosis type 2 (NF2) is an autosomal dominant heritable neoplasia syndrome. It results from a germline mutation of the Despite the significant audiologic morbidity identified in patients with NF2, the mechanisms by which the CVS causes hearing loss are not understood. The most frequently cited hypothesis is the enlarging CVS causes hearing loss through direct compression and stretching of the cochlear nerve. Several phenomena, including the unpredictable onset of hearing loss, frequent association of hearing loss with small tumors and progressive hearing loss in patients with non-growing CVS specifically suggest that such a simple association between tumor size and/or growth rate with hearing loss does not exist Patients enrolled in a National Institute of Neurological Disorders and Stroke (NINDS) institutional review board (IRB)-approved prospective NF2 natural history study (NIH#08-N-0044) were included. Informed written consent was obtained from all adult participants. Informed written consent from the next of kin, carers or guardians on behalf of minors participating in this study was obtained. Both written consent and assent forms were approved for the consenting procedure by the NINDS IRB. Treatment na\u00efve ears with no evidence of conductive hearing loss were used for analysis. Patients had NF2 diagnosed by clinical and/or genetic criteria Detailed histories were obtained and comprehensive otologic and neurologic examinations were conducted in all patients to exclude the presence of middle ear disease. Concurrent audiometric evaluations were performed that included air- and bone- conduction thresholds from 250\u20138000 Hz and 250\u20134000 Hz, respectively. Hearing was stratified utilizing the four frequency pure-tone average (PTA) as normal [PTA\u226420 dB], mild hearing loss [PTA 21\u201340 dB], moderate [PTA 41\u201370 dB], severe [PTA 71\u201395 dB], or profound [PTA>95 dB] Patients underwent magnetic resonance (MR)-imaging with and without contrast (T1 weighted) of the craniospinal axis. A neuroradiologist, blind to the hearing status of individual patients and ears, interpreted all imaging findings. Inner ear MR-imaging was performed with less than 1 mm in plane resolution utilizing a 3T-MR-scanner . Non-contrast enhanced fluid attenuated inversion recovery (FLAIR) sequences were performed to assess for the presence of elevated protein within the inner ear. Because saline-based solutions become hyperintense relative to brain parenchyma at defined protein concentration thresholds, identification of elevated intralabyrinthine protein by pre-contrast FLAIR MR-imaging is dependable and was reported as either being \u201celevated\u201d or \u201cnormal\u201d Volume of the CVS was determined utilizing post-contrast T1-weighted images Statistical analyses were performed as defined in text. A P-value less than or equal to 0.05 was considered significant.Of 100 consecutive patients (200 ears) enrolled into the NF2 natural history study, ears that underwent surgery (84 ears in 58 patients) and/or received radiation therapy (26 ears in 20 patients) for management of cochleovestibular schwannomas were excluded from this study. Additionally, 5 patients (10 ears) receiving chemotherapeutics at the time of this study and 1 patient (2 ears) unable to complete audiometric examination (English second language) were also excluded from evaluation. Three ears with mild hearing loss were also excluded from analysis due to the presence of a conductive hearing loss component by otologic and audiologic evaluation. Therefore, 89 treatment na\u00efve ears in 56 consecutive patients were included. In 23 patients (41%), one ear met criteria for inclusion and in 33 patients (59%) both ears met inclusion criteria. MR-imaging evidence of CVS was present in 84 of the 89 ears; 5 ears had no MR-imaging evidence of CVS. Mean patient age at evaluation was 30\u00b116 years . Forty-nine patients (88%) had audiovestibular symptoms associated with a CVS . Mean agFifty-five ears had normal hearing (PTA\u226420 dB), including 5 (6%) ears without a CVS. Thirty-four ears (38%) had associated hearing loss . Mean PTA was 54\u00b129 dB in the ears with associated hearing loss. Hearing loss occurred gradually over months to years , in a stepwise manner characterized by periodic decrements in hearing associated with intervening periods of stable hearing , in a relapsing-remitting pattern defined by periodic decrements in hearing coupled with partial recovery of hearing during intervening periods , or in a sudden and complete fashion . Subjective hearing loss was not reported in 3 ears (9%) in 3 patients with audiometric evidence of mild hearing loss.3). Other imaging findings and audiovestibular symptoms in the 56 study patients (89 ears) are detailed in Radiographic and audiologic analysis in the 34 hearing loss ears revealed that 32 ears (94% of ears with hearing loss) had elevated intralabyrinthine protein on FLAIR MR-imaging . The 2 hCochlear aperture obstruction by a CVS was closely associated with identification of elevated intralabyrinthine protein in hearing loss ears . While 31 hearing loss ears (91%) with elevated protein in the labyrinth had cochlear aperture obstruction by a CVS, only 1 ear (3%) with elevated intralabyrinthine protein did not have evidence of cochlear aperture obstruction , 2. EarsTwenty-three ears had isolated cochlear aperture obstruction and 8 (26%) had cochlear aperture obstruction associated with either a discrete intralabyrinthine tumor (3 ears) or direcFifty normal hearing ears (91%) harbored a CVS and 5 ears (9%) did not have an associated CVS. Radiographic and audiologic analysis of the 50 normal hearing ears with a CVS demonstrated elevated intralabyrinthine protein in 35 ears and normal intralabyrinthine protein in 15 ears (30%) on FLAIR MR-imaging. The 5 normal hearing ears without CVS did not have evidence of elevated intralabyrinthine protein .Similar to hearing loss ears, cochlear aperture obstruction by a CVS was associated with elevated intralabyrinthine protein in normal hearing ears . While 33 normal hearing ears with elevated intralabyrinthine protein had cochlear aperture obstruction by a CVS, only 2 ears (6%) with elevated intralabyrinthine protein did not have evidence of cochlear aperture obstruction.Specific factors associated with elevated intralabyrinthine protein included isolated cochlear aperture obstruction by CVS , cochlear aperture obstruction with direct labyrinthine invasion by CVS , cochlear aperture obstruction with a discrete intralabyrinthine tumor or unknown mechanism . Endolymphatic hydrops was also detected by MR-imaging in 3 normal hearing ears with elevated protein and block of the cochlear aperture.Twelve normal hearing ears harboring a CVS without elevated intralabyrinthine protein did not have block of the cochlear aperture, including 1 ear (8%) with associated hydrops. Three normal hearing ears (20%) had evidence of cochlear aperture block by CVS without elevated protein.3 (approximately 1.5 cm in maximum linear dimension). While the presence of hearing loss was correlated with larger CVS volume in hearing loss ears , 16 (47%) hearing loss ears had a CVS that was less than approximately 0.5 cm3 .CVSs were identified in 84 ears (94%) and were purely intracanalicular in 38 ears (45%). Mean tumor volume (in 84 ears) was 1.8\u00b14.0 cm 0.5 cm3 . Among 3Previous studies have demonstrated that CVS-associated binaural hearing loss will occur in an unpredictable manner over the lifetime of most NF2 patients The most common imaging findings identified in CVS-associated hearing loss in NF2 was the presence of elevated intralabyrinthine perilymphatic protein and the presence of cochlear aperture obstruction on MR-imaging. Elevated intralabyrinthine protein was found in 94% of hearing loss ears (32 of 34 ears) and in 70% of normal hearing ears (35 of 50 ears). Blockage of the cochlear aperture by an associated CVS was found in 96% of ears (64 of 67 ears) with elevated protein and only in 14% of ears (3 of 22 ears) without evidence of elevated intralabrynthine protein. These findings are consistent with the findings by Silverstein and colleagues The importance that elevated intralabyrinthine protein plays in hearing loss is underscored by previous clinical-imaging studies of the inner ear in other conditions, including sporadic vestibular schwannomas and sudden sensorineural hearing loss (SNHL) The findings of the current study indicate that cochlear aperture block underlies elevation of intralabyrinthine protein. The sensitivity and specificity of cochlear aperture obstruction in identifying elevated perilymphatic protein within the labyrinth was 96% and 87%, respectively. This indicates a pathophysiologic association between cochlear aperture obstruction and the presence of elevated intralabyrinthine protein may exist in NF2-associated CVSs. Based on the results from this study and previous studies examining elevated intralabyrinthine protein in other hearing loss syndromes, NF2-associated tumors may lead to elevated intralabyrinthine protein levels in several ways related to cochlear aperture obstruction that causes abnormal protein clearance and deposition.Elevated intralabyrinthine protein could be the result of cochlear aperture obstruction and disruption of the blood-CSF/labyrinthine barrier caused by permeable tumor vessels Another mechanism by which protein can accumulate within the perilymph associated with cochlear aperture obstruction involves impaired clearance caused by the CVS. It has been shown that the cochlear nerve is integral in the rapid transport of proteins from the cochlea to the cochlear nucleus Elevated intralabyrinthine protein could also be the result of increased protein production or deposition. Analogous to cerebellopontine angle CVSs that are known to cause increased protein content within the circulating cerebrospinal fluid compartment Elevated intralabyrinthine protein on MR-imaging was closely associated with the presence of hearing loss. The importance of elevated intralabyrinthine protein underlying hearing loss is supported by previous studies examining unique perilymphatic proteins that are elevated in CVSs, including \u03bc-Crystallin (CRYM) and low density lipoprotein-related protein 2 (LRP2) A simple association between tumor size and hearing loss does not explain several phenomena. For instance, hearing loss in NF2 has an unpredictable onset and variable progression, which may be gradual, stepwise, relapsing and remitting, or sudden and complete. While previous studies have attempted to link increasing tumor size with hearing loss in patients with NF2, this association has not been established because of the ubiquitous presence of small tumors within all series that are associated with hearing loss. Because hearing loss in NF2 may be caused by end organ degeneration Other pathophysiologic processes of the inner ear structures have been implicated with hearing loss, including the development of endolymphatic hydrops and intralabyrinthine hemorrhage Bevacizumab, initially utilized in NF2 clinical trials for the treatment of growing vestibular schwannomas was associated with hearing improvement in a subset of patients The appropriate stratification of benefit to risk ratio for hearing preservation attempts in NF2 requires delineation of at-risk ears for hearing loss. Because hearing loss is currently the most likely outcome in NF2 ears, it becomes imperative to avoid risk when hearing loss is unlikely to occur. Despite the variable presentation of hearing loss in NF2, most ears with normal perilymphatic protein signal had normal hearing and most ears with hearing loss had elevated perilymphatic protein .Based on the available body of evidence and the current findings that indicate elevated perilymphatic proteins may underlie the final common pathway leading to hearing loss, MR-identification of labyrinthine hyperintensity in normal hearing ears (35 of 50 ears in this study) may serve as a potent imaging biomarker of ears at-risk for developing hearing loss."} +{"text": "Over 25,000 HIV-infected children are currently alive and on highly active antiretroviral therapy (HAART) in India. Limited data are available on the efficacy of treatment and emergence of drug resistance (DR) in pediatric populations. In this study we aimed to characterize the pattern of drug resistance mutations (DRM) in a cohort of perinatally infected children.Blood samples were collected from 61 children who were on first line of therapy for \u2265 6 months. Patients\u2019 demographic and clinical parameters were documented. Viral load was measured using Abbott m2000rt system, Germany. DR Genotyping (using an in-house method) was performed on those with viral load >1000 copies/ml.Among those who had been on ART for a median period of 24 months 51 children (83.6%) achieved virological suppression. There were 10 children with virological failure, and only two amongst these manifested immunological failure. Nine children (90%) had reverse transcriptase-related DRMs, and none had protease inhibitor-related DRMs. The most frequent NRTI mutation was M184V (n=9) followed by two thymidine analogous associated mutations (TAMs) M41L and T215Y (n=2). The most frequent nNRTI mutations were K103N (n=6) and Y181C (n=3).Our study showed a high proportion of children achieving virological suppression with a mean duration of two years of ART. Children with virological failure and drug resistance and without immunological failure will likely promote DRM accumulation and may jeopardise second-line ART options. These data suggest that virological monitoring may help optimise regimen switch in children."} +{"text": "ELN encoding the tissue structural protein elastin +/\u2212Eln mice are hypertensive NCF1 gene as a result of the largest recognized WBS microdeletion\u2014about 1.83 Mb\u2014decreases the risk for hypertension in WBS patients compared to those possessing the more common smaller deletion (1.55 Mb) not incorporating NCF1 ; homozygous deficiency accounts for 20% of patients with chronic granulomatous disease, a disorder associated with repeated infections due to an inability to kill bacteria. p47phox is a major effector of angiotensin II (AngII), as demonstrated by a lack of elevation in blood pressure of Ncf1\u2212/\u2212 mice PLoS Genetics, Campuzano and colleagues replicate the WBS cardiovascular phenotype in a WBS mouse model with and without a deletion of the Ncf1 gene Williams-Beuren Syndrome (WBS) arises when there is a genomic microdeletion at human chromosome 7q11.23 (Mouse 5G2), resulting in various cardiovascular, developmental, metabolic, and mental disorders ing NCF1 , top [5]Limk1 to Trim50 (that contains Eln), manifest a cardiovascular phenotype including hypertension with elevated angiotensinogen (Agt), renin (Ren), and angiotensin converting enzyme (Ace) mRNA throughout life (Gtf2i to Limk1 (not containing Eln) is normotensive independently lowered blood pressure in DD mice and, together, their effect was additive. Blood pressure control was associated with reduced vessel ROS and lowered plasma AngII levels. Further, the anti-ROS therapies reduced the degree of anatomical changes in cardiac hypertrophy and vascular elastic fiber fragmentation. These pharmacologic studies suggest an alternative approach to treating the hypertensive phenotype in WBS patients. Currently, the most common treatments for WBS-induced hypertension are beta-blockers and calcium channel blockers. Using the present WBS mouse studies as an insight, the combined use of a specific antioxidant with losartan appears mechanistically more rational.In addition to elevated AngII levels, DD mice have elevated protein nitrosylation and superoxide anions as determined by dihydroethidium fluorescence in the ascending aorta. In contrast, DD/The use of antioxidants for the treatment of vascular oxidant stress associated with cardiovascular disease has been questioned after failed clinical trials using non-specific antioxidants such as vitamins C and E In addition to hypertension and developmental structural defects in the cardiovascular system, increased vascular ROS may increase arterial thrombosis risk. Several animal models have been associated with increased vascular ROS and higher arterial thrombosis risk. Both heme oxygenase I\u2013deleted mice and prolylcarboxypeptidase-deficient mice have increased vascular ROS and reduced arterial thrombosis occlusion times The studies by Campuzano et al."} +{"text": "Ultrasonic Transcranial Doppler (TCD) of \u201cBIOSS\u201dand\u201cSPECTROMED\u201d companies (Russia),t ranscranial color-coded duplex (TCCD) by \u201cLogic P-5\u201d.All children with headaches were separated according the clinical and ultrasound findings: migraine, tension type of headache, headache with increase or reduction of arterial pressure, headache caused by cerebral venous dysfunction. 30% of them had cerebral anomalies . The children complained of headaches (100%) and also vegetative dysfunction (80%), nasal bleeding (60%), vomiting (40%), dizziness and noise in ears (35%). Venous outflow in the cavernous sinus, straight sinus, great cerebral vein of Galen have been registered by TCD, TCCD. Cerebral venous hemodynamic disturbances (\u201cmarkers\u201d) revealed in all groups children: migraine \u2013 35 %, tension type of headache \u2013 40 %, headache with increase or reduction of arterial pressure \u2013 from 25 % to 55 %. Thus a venous outflow is stimulated and makes influence on intracranial venous circulation. The estimation of cerebral venous hemodynamic disturbances in literature is described mainly in adults, but they are of great importance in clinical manifestations, especially in children. Diagnosis of such disturbances in children is not detected in time, though they often turn out to be one of the main evidence of cerebrovascular pathology. The complex research of cerebral venous hemodynamics presents new possibilities for revealing disturbances of cerebral venous blood circulation. The conservative treatment which has been performed under ultrasonographic control in children with disturbances of cerebral hemodynamics, led to objective improvement in 85% of children."} +{"text": "Recent suggestions to revise guidelines that recommend extending the time for revaccination beyond 10 years may be based on insufficient and conflicting evidence of persistence of immunity did not protect the mice against lethal systemic infection, whereas the highest titer serum (43 U/mL) and the commercial Ig (150 U/mL) protected 50% of the mice. Thus, the levels of residual antibody in vaccinated persons are either not protective or only partially protective in mice. Consistent with the reported protection by the higher levels of antibody, vaccinia immune globulin (VIG), which contains 500 neutralizing U/mL, is effective under some conditions serum from a single-vaccinated study participant containing the low 10 U/mL neutralizing activity (patient A), 2) serum from a single-vaccinated person containing the higher 43 U/mL (patient E), or 3) normal commercial Ig containing 150 U/mL and challenged 24 hours later with one LD"} +{"text": "CMR perfusion (CMRP) imaging using adenosine traditionally requires bilateral arm cannulation. Patients with multiple comorbidities often have difficult venous access and dual cannulation often proves impossible. We used a standard two-way adapter to administer adenosine at a standard rate of 140 mcg/kg/minute over 3 minutes for maximum coronary vasodilatation following a bolus injection of gadolinium. High flow bolus injection may cause sinus arrest caused by a flush of residual adenosine in the same arm vein. We acquired 50 sequential R-wave triggered image frames to assess first pass myocardial perfusion and assessed the effect of significant sinus pauses on image acquisition.2) after administration of a 0.1 mmol/Kg bolus of intravenous Gadolinium (Gadovist). Fifty sequential R-wave triggered frames were acquired to assess first pass perfusion. We assessed the effect of significant sinus pauses on patient safety and quality of image acquisition.First pass perfusion was performed on a Siemens Avanto1.5T MR scanner with a standardised acquisition protocol using intra-venous adenosine 140\u03bcg/kg/min for 3 minutes. Three sequential short axis slices of 8mm thickness were acquired per cardiac cycle using a hybrid EPI sequence developed significant episodes of bradycardia. Mean heart rate 69.1(+/-28.1) Range: 139-18 min-1). Image sequences during first pass perfusion were adversely affected due to gaps in R-wave triggered image frame acquisition during, or just before gadolinium arrival because of the effects of adenosine (Figures Crucial image frames are not acquired at the most important time points during first pass perfusion in 26% of patients with the Octopus Bionector. The introduction of a two-way intravenous adapter resulted in an unacceptably high number of patients having sinus pauses/sinus arrest over several seconds, And as image quality is heavily dependent on R wave intervals being regular, this adversely affected image quality. This could have led to underdiagnosis of perfusion defects in the affected patients.N/A."} +{"text": "The cytoplasmic S100 proteins derived from cells of myeloid origin. Calprotectin (MRP8/14 protein complex) might be a biomarker either for autoinflammation and autoimmunopathy. Since autoinflammatory diseases might be a diagnostic challenge calprotectin may be helpful in the diagnosis of autoinflammatory diseases. Chronic nonbacterial osteomyelitis (CNO) is an autoinflammatory, noninfectious disease. CNO describes a wide spectrum from a monofocal bone lesion to the chronic recurring multifocal osteomyelitis (CRMO). Laboratory and histopathological findings are nonspecific. In some patients systemic inflammatory signs such as elevated acute phase proteins cannot be found.To test the ability of Calprotectin (MRP8/14 protein complex) serum concentrations to monitor disease activity in patients with CNO.Serum concentrations of Calprotectin (MRP8/14 protein complex) in a patient with CNO were determined by a sandwich ELISA.Calprotectin (MRP8/14) level were raised heralding active disease when acute phase proteins . The calprotectin level was 7872,7 ng/ml .Calprotectin (MRP8/14) serum concentrations correlate closely with disease activity and may herald a flare before clinical manifestation. Therefore MRP8/14 serum concentrations are a biomarker indicating disease activity in CNO patients.None Declared."} +{"text": "Sexual selection has been proposed as the driving force promoting the rapid evolutionary changes observed in some reproductive genes including protamines. We test this hypothesis in a group of rodents which show marked differences in the intensity of sexual selection. Levels of sperm competition were not associated with the evolutionary rates of protamine 1 but, contrary to expectations, were negatively related to the evolutionary rate of cleaved- and mature-protamine 2. Since both domains were found to be under relaxation, our findings reveal an unforeseen role of sexual selection: to halt the degree of degeneration that proteins within families may experience due to functional redundancy. The degree of relaxation of protamine 2 in this group of rodents is such that in some species it has become dysfunctional and it is not expressed in mature spermatozoa. In contrast, protamine 1 is functionally conserved but shows directed positive selection on specific sites which are functionally relevant such as DNA-anchoring domains and phosphorylation sites. We conclude that in rodents protamine 2 is under relaxation and that sexual selection removes deleterious mutations among species with high levels of sperm competition to maintain the protein functional and the spermatozoa competitive. The idea that genes involved in reproductive processes evolve rapidly has gained widespread acceptance Since sexual selection is known to drive the rapid evolution of reproductive traits which make sperm more competitive, it has been proposed that it could also drive rapid evolution among genes related to such processes It has been widely assumed that such changes in phenotypic traits would be linked to rapid divergence in the coding sequences of proteins underlying such reproductive processes Mus (Rodentia) found links between the degree of divergence in the promoters of protamine 2, levels of sperm competition, and sperm swimming velocity It has been claimed that protamines are the fastest evolving reproductive proteins and that this is a consequence of sexual selection acting on them Protamines are a diverse family of small, arginine-rich nuclear proteins that replace histones and transition proteins during the process of sperm nucleus condensation in spermatogenesis Expression of protamines is specific to testis and protamine mRNAs are exclusively detected in postmeiotic spermatid stages Protamine 1 is typically 49 or 50 amino acids long and contains a highly conserved arginine rich DNA-binding domain as well as multiple serine and threonine residues that may be used as phosphorylation sites Mus species examined in a previous study . These species represent a more divergent group, which split from other members of the Cricetidae about 18 MYA us study [24], whArvicola sapidus, Arvicola terrestris, Clethrionomys glareolus, Chinomys nivalis, Microtus arvalis, Microtus cabrerae, Microtus agrestis, Microtus gerbei, Pitymys duodecimostatus, Pitymys lusitanicus), 5 to the subfamily Cricetinae , and 1 to the subfamily Sigmodontinae (Sigmodon hispidus)The study includes 16 species of the family Cricetidae, 10 of which belong to the subfamily Arvicolinae ((Prm1) and protamine 2 (Prm2) sequences of Sigmodon hispidus were obtained from NCBI Genebank (Accession EU980395 for Prm1 and EU980396 for Prm2). Prm1 sequence for Phodopus sungorus, Phodopus roborovskii, and Cricetulus griseus, and Prm1 and Prm2 sequences for Mesocricetus auratus were obtained from the literature Protamine 1 Genomic DNA was extracted from different frozen tissues using the E.Z.N.A\u00ae Tissue DNA kit following the manufacturer's recommendations.2 (Roche), 0.8 mM dNTPs mix supplying 0.2 mM of each deoxinucleotide triphosphate , 0.3 mM of forward and reverse primers (Applied Biosystems), 2 U of Taq Gold DNA polymerase (Roche), and 20\u2013100 ng/\u00b5l of genomic DNA template. All PCRs were performed in a Veriti thermocycler (Applied-Biosystems). The conditions of the thermocycler program consisted of 35\u201345 cycles with an initial denaturation of 95\u00b0C for 30\u201340 s, an annealing stage at 52\u201362\u00b0C (depending on template and primers) for 40 s, and an elongation stage at 72\u00b0C for 30\u201350 s (depending on gene length).Protamine sequences were amplified by Polymerase Chain Reaction (PCR). PCR mixtures were prepared in a 50 \u00b5l volume containing PCR Gold buffer 1\u00d7 , 2.5 mM MgClPCR primers were designed on the basis of protamine genomic sequences of other closely related rodent species accessible in the literature or in NCBI GeneBank. All alignments were performed in Bioedit Prm2 and <300 bp size for Prm1 were extracted with E.Z.N.A.\u00ae Gel Extraction Kit (Omega). Purified products were sequenced .PCR products were purified by using the E.Z.N.A.\u00ae Cycle Pure kit (Omega). In cases in which additional nonspecific bands were obtained after separation in a 1.5% agarose gel, bands of <600 bp size for Mus musculus used as outgroup is an indicator of selective pressure at the protein level, with \u03c9\u200a=\u200a1 indicating neutral evolution, \u03c9<1 purifying selection, and \u03c9>1 diversifying positive selection The nonsynonymous/synonymous substitutions rate ratio (\u03c9\u200a=\u200adN/dS) was generated by different models for all/chosen branches of the tree or for each codon site of the alignment . To test for positive selection on branches or sites, the models (see below) are compared with corresponding null-models by means of likelihood-ratio-tests. These tests compare twice the log-likelihoods of the used model and the null-model (conservative) to critical values from a chi-square distribution with the degrees of freedom equal to the difference in the number of parameters between the two models. The positive selection on sites was additionally crosschecked through the program SLR To estimate rates of sequence evolution we used the application Codeml implemented in PAML 4 By computing the clade model comparing Cricetidae as foreground clade against a background of 12 murid species (available from NCBI GeneBank) we obtained the evolutionary rate of the foreground clade in contrast to the background . Three mvs Model 3), Model 3 presents a better fit against the Model 2, and the \u03c9 value estimated is higher than 1. Candidates for relaxation for Cricetidae are reported if two conditions are fulfilled: Cricetidae clade evolves at a significantly different omega than the background (Model 1 vs Model 3), and this omega was not significantly different from 1 (Model 2 vs Model 3) True cases of positive selection for Cricetidae are reported if three conditions are fulfilled: Cricetidae clade and background clade evolve at a significantly different rates of the sum to obtain the \u03c9 value. By calculating \u03c9 in this way we take into account the total accumulated selective pressures in protamines during their evolution, which is more suitable for testing relationships against phenotypic data which do reflect the whole phenotypic evolution To obtain species-specific \u03c9 values to analyse the relation between evolutionary rate and sperm competition levels for each species, we used the free branch model and calculated an \u03c9 value for each species by addition of dWe used the species relative testes mass as a proxy for levels of sperm competition as in our previous studies Species data may not be free of phylogenetic association, since they may share character values as a result of a common ancestry rather than independent evolution, and thus may not be truly independent. To control for this phylogenetic inertia, we used phylogenetic generalized least squares (pGLS) analyses between the \u03c9-values computed from the root and the species relative testes mass. We performed the pGLS analysis using the program COMPARE 4.6b To test evolution along coding sequences and infer amino acids under positive selection we applied likelihood ratio tests comparing a null model that does not allow sites with \u03c9>1 with a selection model that does. We used two likelihood ratio tests. The first compared a nearly neutral model M1a, which assumes values for \u03c9 between 0 and 1, with a model M2a which allows values of \u03c9>1. The second test is more refined and compares two models assuming a \u03b2 distribution for \u03c9 values. In this case, the null model M7 that limits \u03c9 between 0 and 1 is compared to the alternative model M8, that adds an extra class of sites with an \u03c9 ratio estimated that can be greater than 1 Possible target areas of post-translational modifications were determined using the ScanProSite tool of ExPASy Proteomic Server and verified by Net-Phos server 2.0. Proposed phosphorylation sites in the literature DNA-anchoring domains were predicted selecting regions containing 3 or more consecutive arginine or lysine residues flanked by short peptide segments containing cysteine residues, following the literature on structural and functional characteristics of protamines Post-translational processing cleavage sites in the protamine 2 precursor were identified based on previous studies Mus musculus protamines and their domains using the sequence alignment ClustalW implemented in Bioedit A pairwise percentage identity analysis of the amino acid sequences of 5\u2032-AGGAGCAGGGGCAGGGGCAAGGGCTGAG-3\u2032) approximately between bases 70 and 100. This element was found in BLAST searches throughout the mouse genome as well as in virus genomes .NCBI BLAST (Tool: blastn) On the basis of the structural and functional similarities between PRM1 and mature-PRM2, as well as the fact that cleaved-PRM2 shows few similarities to either of them and seems to have a different origin, subsequent analyses were carried out on cleaved-PRM2 and mature-PRM2 separately.Prm1 and the two Prm2 domains suggests a significant difference in the selective constraints of the Cricetidae clade and the rodent background for both domains . The liklaxation .Prm1 showed no significant relationship with relative testes mass although there is a weak negative relationship \u200a=\u200a\u22120.18 to 0.08, correlation\u200a=\u200a\u22120.194) \u200a=\u200a\u221211.54 to \u22122.40, correlation\u200a=\u200a\u22120.69; mature-Prm2: \u03b1\u200a=\u200a8.17, CI 95% (slope)\u200a=\u200a\u22121.98 to \u22120.21, correlation\u200a=\u200a\u22120.574) . In cont\u200a\u22120.574) , Table 2To test selective pressures influencing protamine sequences at the site level, we set a Bayesian Empirical Bayes (BEB) that classifies all sites in three classes in terms of its posterior mean \u03c9. The class differentiation is computed by model 2a or 1a depending on which model shows the best fit (likelihood-ratio test). Class 1 contains sites with \u03c9 values between 0 and 1 and therefore subject to purifying selection. Class contains 2 sites evolving neutrally with an \u03c9 value close to 1. In class 3 (just in model 2a) sites with \u03c9 greater than 1 are indicative of positive selection or functional relaxation. The percentage of sites in each class is shown in The two protamines contain several putative DNA-anchoring domains composed of three or more consecutive arginine and lysine residues Both protamines undergo post-translational modifications through phosphorylation of several motifs containing typically serine and threonine residues. This could regulate the interaction with DNA Phosphorylation sites in cleaved-PRM2 seem to evolve under high selective constraints showing 100% of sites with a low \u03c9 value (class 1). Moreover, evidence of positive selection was not found for any site . In the Precursor-PRM2 is processed after translation by several specific cleaving sites distributed along the cleaved-PRM2 region by a proteolytic process Prm2 sequence of Cricetulus griseus with that of P. sungorus, A. terrestris and M. musculus, representing Cricetidae and non-Cricetidae respectively. The coding sequence of C. griseus Prm2 reveals a complete divergence in relation to its orthologue, suggesting that Prm2 in this species has accumulated successive and defective mutations through evolution thus becoming a dysfunctional gene or pseudogene was found. In contrast, cleaved-protamine 2 and mature-protamine 2 are under relaxation, and there is a negative relationship for both between rate of divergence and relative testes size, suggesting a strong influence of sperm competition.MusPrevious studies on the influence of sexual selection on the evolution of protamines found only weak evidence of positive selection on the protamine 2 precursor in a group of closely related species of It is generally assumed that post-copulatory sexual selection promotes positive selection by driving rapid changes in reproductive genes which result in adaptive advantages It is noteworthy that sexual selection has only been shown to influence proteins belonging to families such as SEMG2 Cricetulus griseus) has a protamine 2 sequence which is highly divergent and mature-protamine 2 is not expressed in sperm. Similarly, other species of this family (Cricetidae) do not present mature-protamine 2 in sperm Cricetulus and Cricetus, do not have mature-protamine 2 in fully-differentiated spermatozoa, while those belonging to the genera Mesocricetus and Phodopus do. Furthermore, among the latter species the proportion of mature-protamine 2 in sperm ranges between 33 and 50%, which is lower than the proportion found in other rodents such as MusCricetulus griseus has become a dysfunctional gene or a pseudogene, supporting the idea that the role of protamine 2 is secondary in this group and that protamine 1, which is present in all species, performs the main function of DNA compaction.The idea that protamine 2 in this group of rodents may suffer from relaxation of selective constraints and degeneration is supported by the fact that one of the species studied and weaker evidence for one additional site (position 38), in agreement with previous findings on rodents Mus found evidence of one site under positive selection In contrast, cleaved-protamine 2 and mature-protamine 2 seem to evolve neutrally. Among cleaved-protamine 2 most sites are under purifying selection, while among mature-protamine 2 most sites are evolving neutrally. No positively selected sites were detected. So far, few studies have analysed the evolution of protamine 2 and they have analyzed the entire precursor protein without distinguishing its two regions Among protamines, functionally important sites include DNA-anchoring domains, phosporylation sites and cleaving sites. DNA-anchoring domains are only present in protamine 1 and mature-protamine 2, phosphorylation sites are present in both protamines, and cleaving sites are by definition only present in the section of protamine 2 which is sequentially cleaved. Most DNA-anchoring domains are under purifying selection in protamine 1, although a substantial proportion are under positive selection. In contrast, all DNA-anchoring domains are evolving neutrally in mature-protamine 2. Similary, most phosphorylation sites are under purifying selection in protamine 1, although a susbtantial proportion are under positive selection. Among cleaved-protamine 2 all phosporylation motifs are under purifying selection, while all of them evolve neutrally in mature-protamine 2. It is important to point out that two positively selected sites detected in protamine 1 are located in phosphorylation motifs, while one (33C) is located in the flanking region of a DNA-anchoring domain, suggesting that they are the targets of selection. The functional importance of DNA-anchoring domains is obvious, while that of phosphorylation sites is currently less clear. Protamines are phosphorylated as soon as they are synthesized and phosphorylation is required for protamines to bind to DNA In conclusion, among rodents protamine 1 and protamine 2 are under different selective constraints, since protamine 1 is under positive selection while cleaved and mature-protamine 2 are under relaxation. The role of sexual selection in rodents seems to be to halt the degree of degeneration of both domains of protamine 2, which in some species has even become dysfunctional and is not expressed in mature sperm. In contrast, protamine 1 is functionally conserved but shows directed positive selection on specific functional relevant sites such as DNA-anchoring domains and phosphorylation sites. While protamine 1 and mature-protamine 2 have many structural similarities, the cleaved region seems to be of retroviral origin. Our findings suggest that since genes in families have many structural and functional similarities, their evolutionary patterns should be studied jointly in order to understand how selective forces act upon all of them in cases where they may play complementary roles or have functional redundancies.Figure S1Phylogenetic trees.A - Tree of study species (Cricetidae). Input tree for branch and site analyses. Mus m. musculus was used as outgroup. B - Tree of study species (Cricetidae) including 12 rodent species as a background. Oryctolagus cuniculus was used as an outgroup. Input tree for clade analyses. Phylogenetic trees were constructed based on literature .(TIFF)Click here for additional data file.Figure S2PAML codeml clade analysis. Models and analysis employed to detect the mode of selection acting on Protamine 1 and Protamine 2 domains. The employed models were compared by means of Likelihood-ratio-tests.(TIFF)Click here for additional data file.Figure S3Amino acid sequence alignment of Protamine 1. Study species (Cricetidae) represented by abbreviated code: Arvicola sadipus (ASA), Arvicola terrestris (ATE), Clethrionomys glareolus (CGL), Cricetulus griseus (CGR), Chionomys nivalis (CNI), Microtus agrestis (MAG), Microtus arvalis (MAR), Mesocricetus auratus (MAU), Microtus cabrerae (MCA), Microtus gerbei (MGE), Mus musculus musculus (MMU), Phodopus campbelli (PCM), Pitymys duodecimcostatus (PDU), Pitymys lusitanicus (PLU), Phodopus roborovskii (PRO), Phodopus sungorus (PSU), Sigmodon hispidus (SHI). Alignment of Protamine 1 including a histogram showing \u03c9 values (red line) estimated under model M2a in each site with standard error (bars). Arrows in Protamine 1 histogram indicate sites subjected to positive selection according to Codeml (PaML 4) site analysis . Evidence for residue 38 was estimated under model M2a and for residues 33 and 39 under M2a and M8. Note that gaps are removed in the alignment but they were included in analysis.(TIFF)Click here for additional data file.Figure S4Amino acid sequence alignment of cleaved-Protamine 2 Study species (Cricetidae) represented by abbreviated code: Arvicola sadipus (ASA), Arvicola terrestris (ATE), Clethrionomys glareolus (CGL), Cricetulus griseus (CGR), Chionomys nivalis (CNI), Microtus agrestis (MAG), Microtus arvalis (MAR), Mesocricetus auratus (MAU), Microtus cabrerae (MCA), Microtus gerbei (MGE), Mus musculus musculus (MMU), Phodopus campbelli (PCM), Pitymys duodecimcostatus (PDU), Pitymys lusitanicus (PLU), Phodopus roborovskii (PRO), Phodopus sungorus (PSU), Sigmodon hispidus (SHI). Alignment of cleaved-Protamine 2 including a histogram showing \u03c9 values (red line) estimated under model M2a in each site with standard error (bars). Note that gaps are removed in the alignment but they were included in analysis.(TIFF)Click here for additional data file.Figure S5Amino acid sequence alignment of mature-Protamine 2. Study species (Cricetidae) represented by abbreviated code: Arvicola sadipus (ASA), Arvicola terrestris (ATE), Clethrionomys glareolus (CGL), Cricetulus griseus (CGR), Chionomys nivalis (CNI), Microtus agrestis (MAG), Microtus arvalis (MAR), Mesocricetus auratus (MAU), Microtus cabrerae (MCA), Microtus gerbei (MGE), Mus musculus musculus (MMU), Phodopus campbelli (PCM), Pitymys duodecimcostatus (PDU), Pitymys lusitanicus (PLU), Phodopus roborovskii (PRO), Phodopus sungorus (PSU), Sigmodon hispidus (SHI). Alignment of mature-Protamine 2 including a histogram showing omega values (red line) estimated under model M2a in each site with standard error (bars). Note that gaps are removed in the alignment but they were included in analysis.(TIFF)Click here for additional data file.Figure S6Codeml output file (M2) for Protamine 1. Included species of Cricetidae represented by abbreviated code: Arvicola sadipus (ASA), Arvicola terrestris (ATE), Clethrionomys glareolus (CGL), Cricetelus griseus (CGR), Chionomys nivalis (CNI), Microtus agrestis (MAG), Microtus arvalis (MAR), Mesocricetus auratus (MAU), Microtus cabrerae (MCA), Microtus gerbei (MGE), Mus musculus musculus (MMU), Phodopus campbelli (PCM), Pitymys duodecimcostatus (PDU), Pitymys lusitanicus (PLU), Phodopus roborovskii (PRO), Phodopus sungorus (PSU), Sigmodon hispidus (SHI).(PDF)Click here for additional data file.Figure S7Codeml output file (M2) for cleaved-Protamine 2. Included species of Cricetidae represented by abbreviated code: Arvicola sadipus (ASA), Arvicola terrestris (ATE), Clethrionomys glareolus (CGL), Cricetelus griseus (CGR), Chionomys nivalis (CNI), Microtus agrestis (MAG), Microtus arvalis (MAR), Mesocricetus auratus (MAU), Microtus cabrerae (MCA), Microtus gerbei (MGE), Mus musculus musculus (MMU), Phodopus campbelli (PCM), Pitymys duodecimcostatus (PDU), Pitymys lusitanicus (PLU), Phodopus roborovskii (PRO), Phodopus sungorus (PSU), Sigmodon hispidus (SHI).(PDF)Click here for additional data file.Figure S8Codeml output file (M2) for mature-Protamine 2. Included species of Cricetidae represented by abbreviated code: Arvicola sadipus (ASA), Arvicola terrestris (ATE), Clethrionomys glareolus (CGL), Cricetelus griseus (CGR), Chionomys nivalis (CNI), Microtus agrestis (MAG), Microtus arvalis (MAR), Mesocricetus auratus (MAU), Microtus cabrerae (MCA), Microtus gerbei (MGE), Mus musculus musculus (MMU), Phodopus campbelli (PCM), Pitymys duodecimcostatus (PDU), Pitymys lusitanicus (PLU), Phodopus roborovskii (PRO), Phodopus sungorus (PSU), Sigmodon hispidus (SHI).(PDF)Click here for additional data file."} +{"text": "Previously, we reported a six-marker gene set, which allowed a molecular discrimination of benign and malignant thyroid tumours. Now, we evaluated these markers in fine-needle aspiration biopsies (FNAB) in a prospective, independent series of thyroid tumours with proven histological outcome.ADM3, HGD1, LGALS3, PLAB, TFF3, TG) in the needle wash-out of 156 FNAB of follicular adenoma (FA), adenomatous nodules, follicular and papillary thyroid cancers (TC) and normal thyroid tissues (NT).Quantitative RT\u2013PCR was performed up to 0.78 and positive predictive values (PPV) up to 0.84. Two FNAB marker gene combinations allowed the distinction of FA and malignant follicular neoplasia with NPV up to 0.94 and PPV up to 0.86.Significant expression differences were found for We demonstrate that molecular FNAB diagnosis of benign and malignant thyroid tumours including follicular neoplasia is possible with recently identified marker gene combinations. We propose multi-centre FNAB studies on these markers to bring this promising diagnostic tool closer to clinical practice. Nodular thyroid disease is one of the most frequent endocrine disorders with an incidence of up to 50%, depending on the diagnostic criteria, in patients beyond the fifth decade of life . Fine-neADM3/HGD1/LGALS3/PLAB/TFF3/TG) for the distinction of benign and malignant thyroid tumours including follicular thyroid neoplasia with a sensitivity of 91%, a specificity of 100%, a positive predictive value (PPV) of 1.0 and a negative predictive value (NPV) of 0.94 , 93 benign thyroid tumours (40 follicular adenoma (FA) and 53 adenomatous nodules) and 45 surrounding normal thyroid tissues.in vivo in a private practice for endocrinology and nuclear medicine in L\u00fcneburg (n=66) and (ii) ex vivo, immediately after surgical removal of the thyroid tumour in the Department of Surgery in Schkeuditz and the University Hospital Halle (n=45). Fine-needle aspirates were gained for cytological and molecular investigation. Cytological investigation was performed by a local pathologist in the cohort of in vivo FNAB and by a pathologist from our University Hospital in the cohort of ex vivo FNAB. Fine-needle aspiration biopsies were classified as recommended by the NCI Thyroid Fine-needle Aspiration (FNA) State of the Science Conference in 2007 . Informed consent was obtained from all patients. The local ethics committee approved the study.In all, 111 patients undergoing surgery for a thyroid tumour were studied. Fine-needle aspirates of the thyroid nodules were obtained prospectively (i) \u03b2-actin (ACTB) was demonstrated in all samples by RT\u2013PCR. Real-time PCR was performed using intron spanning primers for ADM3 (HGD1 (LGALS3 (PLAB (TFF3 (TG and the house-keeping gene \u03b2-actin (ACTB) as previously described (2 concentrations were optimised to create a one-peak-melting curve (primer sequences and PCR conditions are available upon request).Needle remnants were washed out and transferred into TRIzol reagent . RNA extraction and cDNA synthesis were carried out as previously described in upregulation or downregulation of mRNA expression was calculated as follows:The fold difference . For the molecular discrimination of follicular thyroid tumours, mRNA expression levels in 40 FAs were compared with mRNA expression levels in 10 malignant follicular thyroid neoplasias (including three FVPTC). The Mann\u2013Whitney U-test within the SPPS software was used for statistical analysis of mRNA expression differences and a P-value of <0.05 was defined as statistically significant.\u2018Normal tissue\u2019 corresponds to the surrounding thyroid tissue of tumours assembled in the vs malignant thyroid tumours and between FA vs malignant follicular thyroid neoplasia, cross-validation analysis was performed to calculate the optimal discriminative cutoff value between both entities using SPSS software . Additionally, specificity and sensitivity as well as PPV and NPV and accuracy were calculated for each of these marker genes and marker gene combinations.In case of significant expression, differences between benign \u03b2-actin (ACTB) and thyroglobulin (TG) mRNA expression was confirmed in all FNAB samples and was mandatory for further marker gene analysis. Messenger RNA expression for individual marker genes was demonstrated in 109 (TFF3), 99 (LGALS3), 95 (ADM3), 84 (HGD1) and 67 (PLAB) of 111 fine-needle biopsies obtained from the thyroid tumours and in all 45 samples obtained from normal thyroid tissues. There was no difference in mRNA retrieval or LightCycler PCR (threshold cycles) quantification between fine-needle biopsies obtained in vivo or ex vivo.Presence of n=18) showed significantly lower mRNA expression levels for TFF3 and HGD1 and significantly higher mRNA levels for ADM3 and LGALS3 compared with FNAB of benign thyroid tumours (n=93) .vs benign thyroid tumours, with the remainder showing the same trend for upregulation or downregulation as in our previous study . There were no significant differences between benign thyroid nodules and normal thyroid tissues expression in all investigated genes and gene combinations .LGALS3/HGD1 x PLAB/TFF3 or LGALS3/HGD1 x LGALS3/TG.No improvement in diagnostic accuracy was found when calculating the product from best discriminating gene ratios vs FA was possible by quantification of ADM3 expression, and by application of two ADM3-based gene combinations . Furtherene sets . On the ene sets . Furtherene sets .LGALS3/HGD1, LGALS3/TG, ADM3/ACTB, ADM3/TFF3, ADM3/TG, ADM3/ACTB x ADM3/TFF3) revealed correct detection of a malignant follicular thyroid neoplasia in 8 out of 9 FNAB classified as \u2018follicular neoplasm (FN)/suspicious for FN\u2019. One PTC with a non-diagnostic FNAB was correctly identified by the proposed marker set. In three cases of PTC with benign FNAB, one case was correctly classified as malignant by the proposed marker set, one case was misclassified as false negative and in one case the proposed markers were not detected in the needle remnant and ADM3/ACTB as the best predictor of malignancy (sensitivity 89%) with only one misclassified case per group.Furthermore, we analysed ADM3/HGD1/LGALS3/PLAB/TFF3/TG) can be successfully applied for a molecular discrimination of benign and malignant thyroid tumours including follicular thyroid neoplasia can be justified since FNAB from FVPTC evoke the same cytopathological difficulties and uncertainties as FNAB from FA and FTC (utations .LGALS3/HGD1) and a sensitivity of up to 82% (LGALS3/TG) emphasises that molecular analysis of FNAB is a promising tool to improve the differential diagnosis of thyroid nodules (ADM3/ACTB x ADM3/TFF3 provided a two-gene marker tool with a high NPV (0.94) and diagnostic accuracy (0.84) for the exclusion of follicular thyroid malignancy.In this series, we largely confirm our previous results. Thus, we found statistically significant expression differences in 5 out of 7 previously proposed two-marker gene combinations and demonstrate their usefulness for the molecular discrimination of FNAB from benign and malignant thyroid tumours. Using a combination of at least two-marker gene sets, a specificity of up to 87% was much lower in this prospective study than in our previous investigation.TFF3 mRNA as a promising differentiation marker in FNAB of thyroid neoplasia , the classifier used by An important advantage of TFF3 mRNA analysis is that it can serve as an appropriate internal control mRNA, since its expression is restricted to thyroid epithelial cells and cannot be found in blood cells ) and/or patients with accompanying risk factors should be generously referred to surgery after FNAB. Additionally, thyroid nodules presenting with ultrasound criteria, for example, based on the TIRADS classification (However, the implementation and validation of such algorithms can only be achieved by future joint research efforts of multiple thyroid centres with the conviction that molecular FNAB diagnosis should become a supplementary tool in future management of thyroid nodular disease, which could ultimately save patients from unnecessary thyroid surgery and favourably impact thyroid disease-related health care costs."} +{"text": "Mannose binding lectin (MBL) is the activator of the lectin complement pathway. After cerebral ischemia it has been shown that MBL could be a mediator of secondary brain damage, in contrast after traumatic brain injury (TBI) there are data suggesting that it could be linked to neuroprotection. We tested the hypothesis that MBL is involved in the pathophysiology of TBI. We characterized (1) the temporal activation of MBL and (2) the effects of its inhibition in a model of experimental TBI.(1) Male C57/Bl6 mice were subjected to intraperitoneal anesthesia followed by the controlled cortical impact brain injury model of experimental TBI (injury parameters: velocity of 5 m/second and 1 mm depth of deformation). MBL immunostaining was evaluated at various time points after TBI: 30 minutes, 1, 6, 12, 24, 48, 96 hours and 1 week using anti MBL-A and MBL-C antibodies (n = 3). (2) The effects of MBL inhibition were evaluated by comparing functional and histologic outcomes in C57/Bl6 mice (WT) and in MBL knockout (-/-) mice. Functional outcome was tested using the Composite Neuroscore and Beam Walk test weekly up to 4 weeks postinjury (n = 11). Histologic outcome was evaluated by calculating the contusion volume at 4 weeks postinjury (n = 6). Sham-operated mice received identical anesthesia without brain injury.-/-) mice showed neurological motor deficits up to 4 weeks postinjury when compared to their sham controls. Notably, MBL (-/-) mice showed attenuated behavioral deficits when compared to their WT counterpart at 2 to 4 weeks postinjury . In contrast we observed similar contusion volumes at 4 weeks postinjury .We observed a robust MBL-positive immunostaining in the injured cerebral cortex starting at 30 minutes postinjury and up to 1 week, suggestive of an activation of this pathway following TBI. MBL was observed both at endothelial and tissue levels. Consistently, injured WT and MBL (We observed that: (1) MBL deposition and/or synthesis is increased following TBI; and (2) MBL deficiency is associated with functional neuroprotection, suggesting that MBL modulation might be a potential therapeutic target after TBI."} +{"text": "High-throughput screening for interactions of peptides with a variety of antibody targets could greatly facilitate proteomic analysis for epitope mapping, enzyme profiling, drug discovery and biomarker identification. Peptide microarrays are suited for such undertaking because of their high-throughput capability. However, existing peptide microarrays lack the sensitivity needed for detecting low abundance proteins or low affinity peptide-protein interactions. This work presents a new peptide microarray platform constructed on nanostructured plasmonic gold substrates capable of metal enhanced NIR fluorescence enhancement (NIR-FE) by hundreds of folds for screening peptide-antibody interactions with ultrahigh sensitivity. Further, an integrated histone peptide and whole antigen array is developed on the same plasmonic gold chip for profiling human antibodies in the sera of systemic lupus erythematosus (SLE) patients, revealing that collectively a panel of biomarkers against unmodified and post-translationally modified histone peptides and several whole antigens allow more accurate differentiation of SLE patients from healthy individuals than profiling biomarkers against peptides or whole antigens alone. Proteomics research has focused on characterizing the structures and functions of proteins and peptides, the basic functional molecules in biological systems, affording valuable information for understanding fundamental biological processes and developing clinical applications High-throughput screening of the reactivity of large numbers of peptides towards protein targets has been performed using various techniques, including peptide microarrays, which are especially well-suited for screening biomolecules interactions in parallel, given their high-throughput capability in situ synthesis of peptides directly onto substrates and robotic spotting and immobilization of pre-synthesized peptide onto microarray chips et al. recently developed an on silico synthesized peptide microarray for peptide screening down to single amino acid resolution in situ synthesis method, spotted peptide arrays can be fabricated rapidly using pre-synthesized and well characterized peptides, as was demonstrated in the human epigenome peptide microarray platform (HEMP) Previous approaches to peptide microarrays include Due to the relatively low peptide-target binding affinity and a broad span over five orders of magnitude, Here, we present a new peptide microarray platform on non-continuous, nanostructured plasmonic gold films with enhanced NIR fluorescence detection for vastly improving the sensitivity of high-throughput peptide-antibody screening. The gold platform utilizes spontaneously adsorbed avidin for immobilization of biotin-conjugated peptides and biotinylated branched PEG stars to minimize non-specific binding (NSB) background signal. A proof of concept peptide array composed of biotinylated unmodified and post-translationally modified histone peptides was first demonstrated on gold for detecting commercial antibodies with three orders of magnitude improvement in sensitivity over peptide arrays on glass, with the LOD down to the 10 femto-molar (pg/mL) range. The high sensitivity of the peptide array on gold allowed integration with antigen array on the same chip for the first time for simultaneous probing of peptide-antibody and antigen-antibody interactions over a broad dynamic range. Unmodified and post-translationally modified histone peptides together with whole antigens were arrayed on a plasmonic gold substrate for sensitive profiling of antibodies in the sera of systemic lupus erythematosus (SLE) patients vs. healthy controls. A combined set of peptides and antigens was found to give more accurate differentiation of SLE sera from normal sera than peptides or antigens alone, opening the possibility of gold supported peptide-antigen arrays for disease diagnostics.The synthesis of plasmonic gold substrate containing densely packed Au nano-islands capable The NIR fluorescence enhancement phenomenon With the avidin coated plasmonic gold substrate, we performed multiplexed peptide immobilization by robotic spotting of biotinylated small peptide molecules to form a peptide array . Strong th lysine residue acetylated (H2B K20Ac), histone H3 peptide composed of 21 amino acid at the tail region of histone H3 protein, and H3 peptide with the 18th lysine acetylated (H3 K18Ac).Having generated and optimized peptide microarrays on gold, we probed histone peptide-antibody interactions using fetal bovine serum (FBS) solutions spiked with commercial antibodies. Histones are major components of chromatin and also play important biological roles in gene regulation. Post-translational modifications of the N-terminal domains (tail regions) of histone protein/peptides, such as acetylation, phosphorylation, and citrullination are involved in disease development and serve as epigenetic markers Serial dilutions of a mixture of two commercial antibodies against H2B K20Ac and H3 K18Ac each at 1 nM to 10 fM concentration were used to probe the histone peptide microarray followed by secondary antibody-IRDye800 detection . We obseNext, we investigated plasmonic gold peptide-antigen microarrays for profiling autoantibodies in human serum samples. Systemic lupus erythematosus (SLE) is the second most common rheumatic autoimmune disease in the United States and diagnosis is complex and includes clinical and laboratory criteria We constructed peptide-antigen microarrays on plasmonic gold and glass substrates with a panel of 20 histone peptides as well as 6 whole antigens to characterize autoantibodies in the sera of 20 SLE patients . Whole aThe high sensitivity afforded by NIR-FE render integrated peptide and antigen arrays on plasmonic gold useful for profiling antibodies in SLE patients vs. healthy control sera. We profiled peptide interactions with antibodies in serum samples from two groups of subjects, 20 clinically confirmed SLE patients and 20 healthy controls. Serum IgG antibody reactivity against 20 unmodified and modified peptides, together with 6 whole antigens , was obtained for each individual. For a subset of the histone peptides and most of the antigens, we found a clear increase of antibody reactivity of SLE patients compared to healthy control see \u2013S7. Othein situ synthesis onto cellulose membrane (SPOT synthesis) In situ synthesis of peptide microarrays provides a high-density, well-controlled array format for high-throughput binding event analysis. Spotting peptides onto solid substrates is fast and flexible, and additionally the quality of the peptide can be well-controlled. Due to the small molecular weight and variable binding efficiency of peptides, Various methods including high-throughput assays have been developed to investigate peptide-protein, peptide-peptide and protein-protein interactions. Previously, peptide-antibody interaction has been ascertained by ELISA By introducing plasmonic enhanced NIR fluorescence into microarrays, we obtain a new platform for high-throughput screening and probing of biomolecular interactions with high sensitivity. Plasmonic gold film coating on glass is a simple process that affords hundreds of fold NIR fluorescence enhancement. The gold platform is compatible with existing instrumentation used for microarray experiments including printing/arraying and fluorescence scanning. The method of coating plasmonic gold substrates with an avidin layer for immobilization of biotinylated peptides is simple, effective and can be easily performed. The plasmonic microarray platform allows the use of highly diluted serum samples (up to hundreds of fold), useful when sample volumes are limited.There are two fundamental aspects of the highly sensitive peptide microarray platform on gold. The first lies in the physics of plasmonic enhancement of NIR fluorescence by the gold nanostructures, Systemic lupus erythematosus (SLE) is the second most common rheumatic autoimmune disorder affecting 300,000 individuals in the United States, with an annual incidence of 3 cases per 100,000 persons and prevalence of 51 per 100,000 persons th, 12th, 15th or 20th lysine acetylated are reactive to IgG antibody in SLE patients, and the reactivity is higher than that of the unmodified peptide to various degrees (th lysine acetylated) with autoantibodies in SLE patients Our peptide-antigen microarrays on plasmonic gold substrates allow for simultaneous profiling of autoantibodies against modified and unmodified histone peptides and whole antigens, using serum samples derived from SLE patients. Antibody profiling on plasmonic gold substrate was much more sensitive with higher signal to background ratios and broader dynamic range than on glass , allowin degrees . This wath, 12th, 15th, 20th lysine acetylated, Previous histone peptide microarray on streptavidin-glass slide identified several peptides capable of differentiating SLE patient sera and healthy patient sera, including unmodified histone H3 and H2B peptide and H2B peptide with the 5nd and 5th column) or strong reactivity with only a single acetylated H2B peptide (th column and H2B K12Ac in 8th column). We also noticed that some of the SLE sera exhibited obvious reactivity with acetylated H2B peptides but with little reactivity to whole H2B histone antigen (9th row) or unmodified H2B peptide (6th row). This is consistent with previous finding of apoptosis-induced acetylation of histone peptides in the pathogenesis of SLE We observed highly variable IgG antibody reactivity with the histone peptides and whole histone antigens in the sera of the SLE patient group . Some ofWe found that collectively a subset of unmodified and post-translationally acetylated histone peptides in combination with several whole antigens could be used to profile serum antibodies and differentiate SLE patients from healthy individuals with higIn conclusion, we have developed a new peptide-antigen microarray platform on plasmonic gold substrate capable of enhanced NIR fluorescence and background minimization. A model histone peptide microarray afforded hundreds fold NIR fluorescence enhancement and 3 orders of magnitude improvement in sensitivity and dynamic range over commercial streptavidin-glass based peptide arrays. The gold microarray platform is useful for profiling antibodies in human samples to identify peptide and antigen biomarkers, especially those capable of differentiating low abundance or affinity antibodies in SLE patient sera vs. healthy sera. The broad dynamic range allows for integrated peptide and antigen array on the same plasmonic gold chip, affording a powerful approach to profiling human antibodies for better differentiation of disease patients from healthy individuals. Notably, the peptide-antigen/gold platform can be easily adopted for use with existing microarray systems, with little change in procedures and reagents.N-hydroxysuccinimide (NHS) were purchased from Sigma\u2013Aldrich. Ammonium hydroxide (30% ammonia) and hyclone fetal bovine serum were purchased from Fisher Chemicals. Streptavidin coated glass slides were purchased from Arrayit. Unmodified streptavidin was purchased from Jackson Immunoresearch. IRDye800-NHS ester was purchased from Licor Biosciences. 6-armed poly(ethylene glycol)\u2013amine was purchased from SunBio, Korea. Unmodified and modified biotin conjugated peptides were ordered from Keck Biotechnology Resource Laboratory. Goat anti human IgG antibody was purchased from Vector Lab. Histone protein was purchased from Immunovision. Monoclonal mouse anti H2B K20Ac, and H3 K18Ac antibody was purchased from Abcam. Histone antigens and dsDNA were purchased from ImmunoVision. U1\u201370 was purchased from CPC scientific.Superfrost Plus glass slides and quartz slides were purchased from Fisher Scientific and cleaned with acetone, isopropanol (IPA), and methanol. Avidin, chloroauric acid trihydrate, hydroxylamine HCl, sodium borohydride, cysteamine, 1-ethyl-3-(3 dimethylaminopropyl) carbodiimide (EDC), and SLE patients and control serum samples were obtained under Stanford University Institutional Review Board approved protocols and with informed consent (Protocol #14734). Written consent was provided by participants in this study.4 followed by adding ammonium hydroxide at 20 \u03bcL ammonium hydroxide per mL of HAuCl4 solution with rapid shaking for one minute. The slide was washed twice with DI water to remove unbound gold ions and immersed into 1 mM NaBH4 to reduce gold clusters on the glass slide to gold nanoparticle seeds. After further washing twice with water, the slides were incubated in a solution of HAuCl4 and hydroxylamine at a 1\u22361 ratio at 750 \u03bcM each and uniformly shaken for 5 min. The slide was then soaked in 1 \u03bcM avidin in PBS solution at 4\u00b0C overnight, rinsed with PBS and then water.Plasmonic gold substrates were synthesized by immersing glass slides into a solution of 3 mM HAuClScanning electron micrographs were acquired on an FEI XL30 Sirion SEM with FEG source at 5 kV acceleration voltage. XPS was performed with a PHI VersaProbe Scanning XPS Microprobe.Biotin-NHS ester was dissolved in dry dimethyl sulfoxide (DMSO), then mixed with branched PEG-amine in PBS at a 1\u22361 mole ratio and incubated at room temperature for 2 h following excess biotin removal with a G-25 NAP-5 columns .The avidin coated gold slides were loaded into a microarray printing robot (Bio-Rad) where 0.2 mg/ml biotin conjugated peptide in PBS supplemented with 4% glycerol was printed using solid pins (Arrayit) at 25\u00b0C and 60% humidity, resulting in microarray feature diameters of \u223c200 \u03bcm. The microarray layout is designed by the microarray printer software. A 4-plex peptide array was constructed in a 4\u00d75 matrix with one peptide on each row in replicates of 5 . The sliPeptide and antigen printing and blocking procedure was the same as above. 20 \u03bcL of 1\u2236200 dilution of human serum in PBS solution with 20% FBS was applied to the array for 2 h, followed by washing twice with PBST, once with PBS. 1 nM IRDye800 labeled goat anti human IgG in PBS solution with 20% FBS was applied to each array set subsequently for 20 min in dark. After that, the slide was washed twice with PBST, once with PBS, once with deionized water and dried with compressed air.The commercial Licor Odyssey scanner was applied to scan the peptide microarray assays on different substrates with the 800 nm channel. Genepix 6.1 was used to automatically identify features above composite pixel intensity of 5. Fluorescence intensity for each set of features was background corrected average of mean pixel intensity values for features printed in replicates. Multi-experiment viewer was used to implement significance analysis of microarray for the human sera profiling result based on multiplexed peptide microarray. The SAM algorithm was applied to numerical MFI (mean fluorescence intensity) values. Peptide and antigen reactivity that significantly correlated with SLE classes was determined by 1,000 permutations of repeated measurements to have a false discovery rate of 0 . Autoantibody reactivity heatmaps were generated using Multiexperiment Viewer using average linkage Euclidean distance hierarchical clustering.Figure S1Fluorescence enhancement dependence on the number of protein layers. A) For the \u2018avidin IR800\u2019 column: Mean IRDye800 fluorescence signal for a monolayer of IRDye800 labeled avidin adsorbed on plasmonic film and glass slide. Both the plasmonic film and glass slide were soaked in an IRDye800 labeled avidin solution at 4\u00b0C overnight and then rinsed with water prior to fluorescence intensity measurement with a Licor Odyssey scanner. For the \u2018avidin-biotin BSA-avidin IR800\u2019 column: signals measured on 3 layers of avidin \u2013 biotinylated BSA \u2013 IRDye800 labeled avidin on plasmonic film and glass slide respectively. Both the plasmonic film and glass slide were soaked in avidin at 4\u00b0C overnight, after washing with twice PBST and once PBS, the slides was soaked in biotin conjugated BSA solution for 1h, followed by washing with PBST twice and PBS once and incubation in IRDye800 labeled avidin for 1h. Fluorescence intensity was checked by Licor Odyssey scanner. In this case, the IR800 labeled avidin is on the third layer of the protein stack. B) Fluorescence enhancement factor for 1 layer and 3 layer structures based on the data in A), suggesting no significant fluorescence enhancement loss when fluorophore is two protein layer away from the plasmonic substrate.(TIF)Click here for additional data file.Figure S2Peptide microarray profiling with different blocking reagents. The avidin coated gold slides were loaded into a microarray printing robot (Bio-Rad) where 0.2 mg/ml biotin conjugated peptide H2B K12Ac, H3 K18Me1, H2B K5Ac, H2B (aa1\u201321) were printed in 4 rows with triplicates. The slides were dried in a desiccator and then blocked in 200 \u03bcM biotin conjugated PEG-star, biotin conjugated straight PEG chain or biotin only for 20 min, followed by washing twice with PBST and once with PBS. The microarray was probed with SLE serum sample and detected with IRDye800 labeled antihuman IgG antibody. A) Fluorescence images for SLE patient serum probed on avidin/gold slide with biotin-PEG star blocking, linear biotin-PEG blocking, and biotin blocking respectively. B) Corresponding background and spot signals for the three blocking methods in (A). The lowest background was detected with biotin-PEG stars, which facilitated higher signal/noise ratios and peptide arrays with high sensitivity and broad dynamic ranges.(TIF)Click here for additional data file.Figure S3Box plot of serum IgG antibody reactivity from 20 SLE patients and 20 healthy controls against unmodified and modified histone H2A peptides.(TIF)Click here for additional data file.Figure S4Box plot of serum IgG antibody reactivity from 20 SLE patients and 20 healthy controls against unmodified and modified histone H2B peptides.(TIF)Click here for additional data file.Figure S5Box plot of serum IgG antibody reactivity from 20 SLE patients and 20 healthy controls against unmodified and modified histone H3 peptides.(TIF)Click here for additional data file.Figure S6Box plot of serum IgG antibody reactivity from 20 SLE patients and 20 healthy controls against unmodified and modified histone H4 peptides.(TIF)Click here for additional data file.Figure S7Box plot for SLE patient and healthy control sera IgG antibody reactivity against whole antigens including histone H2A, H2B, H3 and H4 proteins, U1\u201370 and dsDNA.(TIF)Click here for additional data file.Table S1Amino acid sequences of printed histone peptides in the peptide-antigen arrays. Ac: acetylated; aa: amino acid; Me1: methylated; Me2: dimethylated; Me3: trimethylated; Ph: phosphorylated; K: Lysine; S: Serine. Number indicates amino acid position from the N-terminus of its corresponding histone proteins.(TIF)Click here for additional data file.Table S2q- and p-values for peptides and antigens included in the peptide-antigen microarray platform in differentiating SLE patient and healthy control groups derived from Significance Analysis of Microarray (SAM).(TIF)Click here for additional data file."} +{"text": "In vivo, cortical pyramidal cells are bombarded by asynchronous synaptic input arising from ongoing network activity. However, little is known about how such \u2018background\u2019 synaptic input interacts with nonlinear dendritic mechanisms. We have modified an existing model of a layer 5 (L5) pyramidal cell to explore how dendritic integration in the apical dendritic tuft could be altered by the levels of network activity observed in vivo. Here we show that asynchronous background excitatory input increases neuronal gain and extends both temporal and spatial integration of stimulus-evoked synaptic input onto the dendritic tuft. Addition of fast and slow inhibitory synaptic conductances, with properties similar to those from dendritic targeting interneurons, that provided a \u2018balanced\u2019 background configuration, partially counteracted these effects, suggesting that inhibition can tune spatio-temporal integration in the tuft. Excitatory background input lowered the threshold for NMDA receptor-mediated dendritic spikes, extended their duration and increased the probability of additional regenerative events occurring in neighbouring branches. These effects were also observed in a passive model where all the non-synaptic voltage-gated conductances were removed. Our results show that glutamate-bound NMDA receptors arising from ongoing network activity can provide a powerful spatially distributed nonlinear dendritic conductance. This may enable L5 pyramidal cells to change their integrative properties as a function of local network activity, potentially allowing both clustered and spatially distributed synaptic inputs to be integrated over extended timescales. In the brains of awake animals, networks are active even when there is no input from the outside world. Neurons embedded within cortical networks experience this intrinsic ongoing firing as \u2018background\u2019 synaptic input. While the effect of this background input on the integration properties of neurons has been studied in the cell body region, little is known about how asynchronous background activity affects integration in distal dendrites, which contain nonlinear mechanisms that boost and dampen synaptic input. Our simulations, using a model of a cortical L5 pyramidal cell, show that the nonlinear NMDA receptor conductance activated by distributed background activity could increase the gain of the dendrite, enabling synaptic inputs to be integrated more effectively over the dendritic tree and over longer time intervals than previously thought possible. This mechanism could potentially enable the integrative properties of individual neurons to change as a function of the activity of the network in which they are embedded. Our work suggests that background network activity could play a key role routing and transforming information as it flows through the cortex. Pyramidal cells are the principal excitatory neurons in the cerebral cortex and those in layer 5 (L5) form its primary output In vivo, cortical neurons are constantly bombarded with synaptic activity In vitro studies show that voltage-dependent synaptic NMDA receptors (NMDARs) can sustain local regenerative responses called NMDAR spikes in the fine dendrites of pyramidal cells 2+ spike in the apical dendrite in vivo experiments show that synaptic input evoked by sensory stimuli are dispersed over the dendritic tree of pyramidal cells in visual in vivo, where inhibition is stronger in vitro conditions where inhibition is often reduced or absent. Nevertheless, NMDAR spikes have been reported in L4 spiny stellate neurons of barrel cortex following whisker stimulation 2+ transients have been recorded in the tufts of L5 pyramidal cells when sensory input occurs during motor activity, but evidence suggests that Ca2+ spikes rather than NMDAR spikes were responsible in vivo, but it is unclear to what extent nonlinear NMDAR conductances contribute to synaptic integration in L5 pyramidal cells and what spatio-temporal patterns these cells respond to in vivo.in vivo-like background network activity on synaptic integration in L5 cortical pyramidal cells using a morphologically and biophysically detailed model that reproduces a wide range of experimentally measured synaptic, dendritic and somatic behaviours in vivoWe have investigated the potential impact of ). Groups of synaptic inputs were generated with the software neuroConstruct in vitro experimental results. However, to understand the role of background input under more physiological conditions, we then examined the properties of pyramidal cell integration when background excitation and inhibition were balanced, as observed in vivo.To investigate how background network activity influences synaptic integration in tufted L5 cortical pyramidal cells, we adapted an existing model consisting of a detailed morphology Fig. 1A in vitro conditions used to study synaptic integration with glutamate uncaging, where background activity is reduced due to long range inputs being severed in acute slices and inhibition is blocked by MNI-glutamate Fig. S1). By stochastically varying the spatial and temporal patterns of synaptic input onto a dendritic branch, and measuring the occurrence of NMDAR spikes over 10 trials, it was possible to build an input-output (I-O) relationship for each individual branch. That is, the relationship between the number of stimulated synapses and the probability of generating an NMDAR spike (P(NMDAR spike)). \u22121We examined how many synapses onto an individual terminal dendritic branch in the apical tuft were required to trigger a regenerative NMDAR event in the absence of background synaptic input. This condition roughly approximates the in vivo from both anatomical and functional measurements. Tufted L5 pyramidal cells are innervated by \u223c15,000 excitatory synaptic contacts (assuming 1 per spine), of which 3500 occur on the apical trunk and dendritic tuft in layers 1\u20133 in vivo2+ hot zone; Blue region, We next examined how asynchronous background excitatory synaptic input affected integration in the tuft. We did not include background inhibition in these initial simulations, in order to study the effects of background excitatory inputs in isolation. We estimated the rate of background excitatory synaptic input We next examined how the branch I-O relationship was affected by background excitatory synaptic input. These simulations revealed that many fewer coincident synaptic inputs were required to trigger an NMDAR spike than for quiescent control conditions Fig. 1G.To investigate whether background excitatory synaptic input also affects the spike output of the pyramidal cell model, we determined the number of stimulated branches required to trigger somatic APs. To do this we randomly selected a group of N branches within our set of 28 terminal branches in the tuft and simultaneously stimulated each branch with 30 near-synchronous synaptic inputs (5 ms window) to ensure a high probability of triggering an NMDAR spike in each branch under all conditions Fig. 1H.By contrast, in the presence of background excitatory synaptic input, the relationship between the probability of triggering APs (P(AP)) and the number of stimulated dendritic branches exhibited a lower activation threshold and was much steeper burst of synaptic input, but activated different branches at random times within a time window that ranged from 5 ms to 200 ms and examined how desynchronization of branch activation affected the probability of triggering APs. In the absence of background excitatory input, 4 stimulated branches produced only a subthreshold response Fig. 3A1By contrast, background excitatory synaptic input substantially reduced the number of stimulated branches required to trigger APs cell-like mixed GABAA/GABAB synapses onto the apical dendritc tree. These inputs were driven at firing rates reported for passive touch whisker experiments in awake animals and are likely to reflect the upper end of the rates observed during active touch, which fall to about half this level on average A receptor component had a fast rise and a decay time of 10 ms B receptor-mediated K+ conductance had a slow rise and decay time of 50 and 80 ms, respectively, with peak values of 0.5 nS and 0.06 nS and an activation delay of 10 ms ). This produced somatic IPSPs with properties comparable to those measured experimentally Fig. S2). Although the inhibitory synapses were fewer in number than the excitatory synapses, their higher firing rate and slower kinetics effectively counterbalanced the AMPAR component of the 1500 excitatory synapses, producing a time averaged GABAR/AMPAR conductance ratio of 1.5, comparable to that estimated from synaptic currents To examine how synaptic integration in L5 pyramidal cells might operate under more physiological conditions, when network activity delivers balanced excitatory and inhibitory background synaptic input, we added 60 SOM-like GABAA receptor mediated conductance, suggests that the slow low-amplitude GABAB component and the GABAA component are equally effective at inhibiting NMDAR spikes mediated inhibition, the NMDAR spike threshold was reduced compared to quiescent conditions, but not as strongly as during background excitation alone Fig. 5A.A and GABAB receptor-mediated inhibitory conductances can counteract the effects of background excitatory input on synaptic integration in the dendritic tuft by raising NMDAR spike threshold and lowering the dendritic gain. Our simulations predict that during periods of lighter dendritic inhibition, for example when both sensory and motor systems are engaged For spatially distributed synaptic input, the number of nearly synchronous inputs required to trigger APs was only marginally higher for 2\u00d7SOM-only balanced background inhibition than background excitation alone Fig. 5B2In the subsequent sections we investigate the mechanisms underlying network activity-dependent spatio-temporal integration in L5 pyramidal cells.Several dendritic mechanisms could be involved in the network activity-dependent changes in synaptic integration we observe, but two are likely to be particularly important. Background excitatory inputs activate glutamatergic synapses that: 1) depolarize the dendrite Fig. 1F Fig. S3A1) that depolarized the apical tree to a level comparable to that during background excitatory synaptic input . Current injections accounted for a substantial part of the change in the single-branch I-O relationship observed with background input . However, when spatially distributed AMPAR-only synapses were used to depolarize the apical tree to a comparable level of depolarization obtained with 1500 AMPAR/NMDAR synapses, they were less effective at altering the branch I-O relationship, due to the shunt introduced by these conductances . Similarly, AMPAR-only background activity that generated comparable dendritic depolarization only accounted for a fraction of the changes observed in the neuronal I-O relationship in response to clustered and spatially distributed inputs . These results show that, while depolarization is important in determining NMDAR spike threshold Since the threshold of NMDAR spikes is voltage-dependent d inputs Fig. 4C.Comparison of the properties of NMDAR spikes in apical branches during different levels of background excitatory synaptic input revealed that the duration of NMDAR spikes systematically increased with the amount of background activity Fig. 6D.; Movie S1). In the absence of background input, triggering an NMDAR spike in branch 11 with 30 nearly synchronous synaptic inputs produced a large voltage depolarization (background\u2212peakcontrol\u226513 mV). The average number of additional activated branches ranged from 0.46 with 900 background excitatory inputs, to 2.19 with 1500 background excitatory inputs (Movie S1). This nonlinear dendritic amplification potentially explains why fewer stimulated branches are required to trigger APs in the presence of background excitatory synaptic input asynchronous background synaptic input lowered NMDAR spike threshold and elongated the duration of NMDAR spikes . Propagation of voltage and activation of spatially distributed NMDAR conductance in neighbouring branches can therefore account for the increased efficacy of NMDAR spikes and the extended spatio-temporal integration we observe in the L5 pyramidal cell model, during background excitatory synaptic input.These results suggest that synaptic AMPARs and NMDARs activated by background excitatory input can mediate active propagation into neighbouring branches, under certain conditions. However, the presence of dendritic NaB receptor inhibition was effective in modulating temporal integration .We next examined the effect of background excitation and inhibition on the spread of NMDAR spikes into neighbouring branches. The average fractional increase in the number of branches exhibiting regenerative potentials was reduced from 2.19 to 1.2 when inhibition was added to background excitation Fig. 9C.in vivo, can extend the spatial and temporal properties of dendritic integration in L5 pyramidal cells. Spatio-temporal integration depends on background network activity, because it introduces spatially distributed synaptic NMDAR conductance over the dendritic tree and depolarizes the dendrites. Increasing the background excitatory synaptic input to levels expected in vivo has three main effects: 1) the number of coincident synaptic inputs required to trigger an NMDAR spike in a branch is reduced; 2) the duration of NMDAR spikes is increased, and 3) NMDAR-mediated regenerative events spread and trigger additional regenerative events in neighbouring dendritic branches. These mechanisms markedly reduce the number of spatially distributed synapses or stimulated terminal dendritic branches required to trigger APs. Inclusion of fast (GABAA) and slow (GABAB) receptor-mediated inhibitory conductances in the background synaptic input in an approximately balanced excitatory/inhibitory configuration only partially counteracted these effects. Our results show that in vivo-like balanced background input could enable L5 pyramidal cells to integrate spatially distributed and/or temporally dispersed synaptic inputs onto the tuft more effectively. Our findings suggest that the activity state of the cortical network can dynamically control the integrative properties of L5 pyramidal cells by adjusting the density of glutamate-bound NMDARs on the dendritic tree. This prediction has important implications for cortical processing.Our simulations suggest that the background synaptic input arising from the ongoing cortical network activity observed +, Ca2+ and K+ channels), which are typically modulated slowly, through plasticity Our simulations show that spatially distributed synaptic NMDAR conductances arising from ongoing network activity extend spatial and temporal integration in the dendritic tuft of L5 pyramidal cells. This network activity-dependent nonlinear dendritic mechanism is distinct from the well-documented effects of noise and linear synaptic conductances on neuronal I-O relationships 2+ below \u221260 mV in vivo-like network activity. Moreover, our control simulations indicate this effect is robust across a range of NMDAR kinetics: the slower the decay of the synaptic NMDAR conductance the longer the absolute window for temporal integration in the absence of other counteracting conductances . However, faster NMDAR kinetics would reduce the impact of background synaptic input unless there was a proportionally higher background input rate or a larger NMDAR/AMPAR peak amplitude ratio to maintain the same time-averaged NMDAR conductance. Another key property of NMDARs that sets them apart from nonlinear dendritic Na+ conductances Several properties of the GluN2A/B containing NMDARs present in L5 pyramidal cells are important for dendritic integration in the presence of background synaptic activity. Their strong block by MgA receptor mediated inhibition to the tuft region in vivo recordings show that SOM cell firing is suppressed for about 1 s during active whisker touch behaviour, suggesting that the tuft region is disinhibited during this behaviour exc/Ginh ratio is higher during in vivo up-states B-mediated inhibition arising from NGF cells By increasing the gain of dendritic integration, background excitation could trigger an unconstrained positive feedback loop, integrating over longer timescales and resulting in prolonged global activation of the dendritic tuft. In practice, cortical microcircuits operate in a balanced configuration with inhibition closely tracking excitation + and Ca2+ conductances clearly aid background induced regenerative activity . Indeed, we found that Ca2+ spikes occur in the tuft during strong tuft stimuli, and help to electrically couple the tuft to the soma, as found experimentally h reduced the NMDAR spike threshold in the presence of background input, but had negligible effect on NMDAR spike duration . Indeed, more complex effects are seen when conductances have activation and/or inactivation properties with significant voltage dependences around \u221255 mV, because background synaptic input depolarizes the dendrite by several millivolts compared to the quiescent condition. For example, transient A-type K+ conductances inactivate strongly over this voltage range making their impact weaker than expected . By contrast, the sustained slow K+ conductance (delayed rectifier) strongly activates during NMDAR spikes and truncates their duration . Increasing the K+ conductances to match levels recently reported experimentally Fig. S7). However, the time course of the resulting dendritic spikes generated under these conditions was markedly different from NMDAR spikes recorded experimentally in L5 pyramidal cells + conductances in the tuft region + conductances appear well placed to modulate spatio-temporal integration in L5 pyramidal cells. SK type channels in spines, which are activated by local Ca2+ influx though NMDARs Fig. S8). Thus the level of expression of the various dendritic conductances and their precise voltage and Ca2+ dependencies are likely to tune the spatio-temporal integration. Irrespective of the natural configuration of the dendritic conductances present in vivo, our results using a passive L5 model, show that the spatially distributed NMDAR conductance arising from ongoing network activity will extend spatio-temporal integration unless other dendritic conductances are specifically configured to counteract this basic property. Indeed, spatially distributed glutamate-bound NMDARs arising from ongoing network activity extend the toolbox of dendritic conductances available to pyramidal cells, potentially enabling them to perform a wider range of behaviours.The dendrites of pyramidal cells contain many different voltage-gated conductances in vivo-like background synaptic input can extend both the spatial and temporal integration in L5 pyramidal cells by providing both depolarization The fine dendrites of pyramidal cells have traditionally been thought to act as coincidence detectors on fast time scales in vivo during network activity than in vitro when the network is largely quiescent. Another important consequence of spatially distributed NMDAR conductance arising from background network activity is that it enables active propagation of NMDAR-mediated regenerative events into neighbouring branches, resulting in a multi-branch regenerative plateau potential. This prediction is consistent with recent in vivo recordings of NMDAR spikes in L2/3 pyramidal cells, which reported that 83% of events occurred in multiple branches in vivo-like levels of background input our L5 model could integrate spatially and temporally dispersed synaptic input. This suggests that spatial clustering of synaptic inputs required for triggering NMDAR spikes under quiescent in vitro conditions in vivo. Hence, while spatial clustering of synaptic input onto the dendrites of pyramidal cells has been found Our simulations show that background synaptic input can lower the number of coincident synaptic inputs required to trigger an NMDAR spike in a distal branch. This suggests that NMDAR spikes are much more likely to occur 2+ influx observed are consistent with a small number of distributed thalamo-cortical inputs being amplified by the dendritic NMDAR conductance arising from local network activity, but alternative scenarios are also possible. In L2/3 pyramidal cells in somatosensory cortex, both spontaneous and hind paw-evoked NMDAR spikes are localized to \u223c30 \u00b5m regions of the dendritic tree and usually encompass multiple branches. These events are effective at triggering APs but do not appear to involve a dendritic Ca2+ spike 2+] signals have been observed in the tuft of L5 pyramidal cells in awake animals performing active whisker touch, when vibrissal sensory input is combined with L1 input arising from the primary motor cortex in vitro when apical trunk Ca2+ spikes were paired with dendritic tuft depolarization or synaptic input. This suggests that in vivo tuft plateau potentials and Ca2+ influx are caused by apical dendritic Ca2+ spikes spreading into the tuft when it is depolarized by L1 input. Although, Ca2+ conductances are clearly important 2+ influx. Indeed, recent in vivo recordings from CA1 pyramidal cells show that both NMDARs and Ca2+ channels are required to generate the slow, widespread regenerative dendritic events that underlie complex spike bursts 2+ channels can set the spatial spread of regenerative dendritic events in pyramidal cells. Our results highlight the role of spatially distributed NMDAR conductances in pyramidal cells under in vivo conditions, by showing how they could enable these cells to integrate both spatially clustered and spatially distributed inputs over longer timescales than previously thought.Widespread NMDAR spikes have been reported in the dendrites of L4 spiny stellate neurons of rat barrel cortex, during whisker deflection + channels 2+ influx in vivo, by faster dynamic modulation conferred by the background synaptic excitation and inhibition arising from ongoing network activity. Thus, both the past and the present activity state of the network determines which spatio-temporal patterns of synaptic input a L5 pyramidal cell can respond to.The finding that synaptic input and nonlinear membrane conductances can produce local regenerative dendritic events has lead to the idea that individual dendritic branches can operate as independent thresholding units Network activity-dependent spatio-temporal integration could have important implications for cortico-cortical and thalamo-cortical signalling. While global network activity is maintained at a relatively constant level due to tight inhibitory control For full methodological details see Supplemental Experimental Procedures.Text S1 and any changes are noted in the figure legends. Simulations were carried out with a fixed integration time step of 0.025 ms and voltages were recorded from the soma and the proximal segment of terminal dendritic branches.The version of the L5 pyramidal cell model gAMPA) component of each quantal glutamatergic synapse was modelled with an instantaneous rise and a single exponential decay:maxg AMPA) was 0.5 nS and the decay time constant (\u03c4decay) was 2 ms The AMPAR conductance (gNMDA) component of each quantal glutamatergic synapse was modelled with an exponential rise and decay, and a voltage-dependence function to mimic Mg2+ block:maxg NMDA is the maximal peak unblocked conductance (1 nS). Time constants for the rise (\u03c4rise) and decay (\u03c4decay) were 3 ms and 70 ms, respectively 2+-dependent inactivation (CDI) of the NMDAR current. This was implemented by multiplying the gNMDA by \u2212he, where the change in h at each time step (h\u2032) is as follows:The NMDAR conductance of 0.5 nS, a \u03c4rise\u200a=\u200a0.3 ms and a \u03c4decay\u200a=\u200a10 ms A is the peak normalization factor as defined above and the Cl\u2212 reverse potential was \u221275 mV. Synapse arising from neurogliaform (NGF) cells, also had a slow GABAB receptor component with a \u03c4rise\u200a=\u200a50 ms and a \u03c4decay\u200a=\u200a80 ms + channels. The maximal peak conductance (gmaxGABA\u200a=\u200a0.06 nS) was estimated from B component accounts for \u223c1/5 of the peak IPSP for NGF cell to L5 pyramidal cell connections. The conductance was obtained by scaling 0.1 nS by the ratio between the driving forces of the GABAA and GABAB components using a K+ reverse potential of \u221287 mV.Quantal GABAergic synapses were modelled with an exponential rise and decay:A receptor mediated synapses, each driven with a 3 Hz random train (2\u00d7SOM configuration) or by a mix of 65 pure GABAA receptor-mediated synapses firing random trains at 3 Hz and 13 GABAA/GABAB receptor-mediated connections (10 synapses each) firing random trains at 14 Hz (SOM+NGF configuration). In both cases this gave a time averaged GABAA receptor conductance of 1.5 times the background AMPAR conductance, comparable to that measured experimentally Two groups of synaptic input were defined. Those that mimicked a stimulus-evoked event and those that mimicked asynchronous background network activity. For simulations where specific dendritic branches were stimulated, one of the 28 terminal branches was randomly selected and the stimulus evoked quantal synaptic inputs randomly placed along it. For spatially distributed stimulus-evoked inputs, quantal synaptic inputs were randomly distributed over the entire apical tuft . NMDAR spikes were identified with a threshold crossing criteria of \u221230 mV . Dendritic branch and neuronal I-O functions were fit using a sigmoid function with the form:n is the rate, 0P is the offset, maxP is the maximum probability of activation and x50 is the number of synapses or stimulated branches at which P(x) reaches half maximum.Voltage traces from the simulations were analysed in IGOR Pro using NeuroMatic software Voltage in randomly selected terminal branches stimulated with 3\u201330 synapses in the absence of background synaptic input. Voltage responses were classified by eye as EPSPs or NMDAR spikes on basis of shape. (B1\u20134) As for (A), but with 1500 background excitatory synapses. (C1\u20134) As for (A), but with balanced background synaptic input . (D) Relationship between decay time and peak depolarization of single events during control. Due to variability in both parameters the populations of EPSPs and NMDAR spikes are partly overlapping. During the 15 synapses stimulation a simple threshold criteria at \u221230 mV (dashed line) misclassified 10% of the NMDAR spikes and 13% of EPSPs (N\u200a=\u200a480). (E) Relationship between decay time and peak depolarization of single events during background excitation. Distributions of EPSPs and NMDAR spikes are reliably separated by the \u221230 mV threshold with only 1% NMDAR spikes classified as EPSPs and none of the EPSPs were misclassified. (F) Relationship between decay time and peak depolarization of single events during balanced background synaptic input. Distributions of EPSPs and NMDAR spikes are reliably separated by the \u221230 mV threshold with 4% of the NMDAR spikes classified as EPSPs and 1% of EPSPs classified as NMDAR spikes.(TIF)Click here for additional data file.Figure S2A and GABAB receptor mediated inhibition.GABA (A) Conductance profile for GABAA receptor synaptic component (red) and GABAB receptor synaptic component mediated by K+ conductance (green). (B) Somatic IPSP produced by a single mixed GABAA/GABAB synapse on a terminal apical branch.(TIF)Click here for additional data file.Figure S3Voltage depolarization alone cannot account for increased probability and efficacy of NMDAR spikes during excitatory background activity. (A1) Cartoon of L5 pyramidal cell indicating the area where currents were injected (red) in order to produce the average depolarization produced by the background activity of 1500 excitatory synapses in the apical tuft. (A2) Probability of NMDAR spike occurrence (P(NMDAR spike)) versus number of stimulated synapses, during control (black dashed line), background excitatory synaptic activity (blue line), apical current injection (red line) and AMPAR-only background synaptic activity reproducing average depolarization and fluctuations (A3 inset). (A3) Probability of action potential P(AP) occurrence versus number of stimulated apical branches (30 synapses per branch) during different conditions. (B1) Cartoon indicating location of current injection to produce the average depolarization produced by the background activity of 1500 excitatory synapses in the Ca2+ spike initiation zone. (B2) P(AP) versus number of stimulated branches, during control (black dashed line) background excitatory synaptic activity (blue line) and Ca2+ zone depolarization (red line). (B3) Absolute decrease in number of stimulated branches required to trigger an AP (P\u200a=\u200a0.5) during apical synaptic input (blue) and Ca2+ zone depolarization. (C1) Cartoon indicating location of current injection to produce the average depolarization produced by the background activity of 1500 excitatory synapses at the soma. (C2) P(AP) versus number of stimulated branches, during control (black dashed line), background excitatory synaptic activity (blue line) and somatic depolarization (red line). (C3) Absolute decrease in number of stimulated branches required to trigger an AP (P\u200a=\u200a0.5) during apical synaptic input and during somatic depolarization.(TIF)Click here for additional data file.Figure S4Effect of changing amplitude and time course of the NMDAR synaptic conductance on NMDAR spikes. (A) Average probability of triggering an NMDAR spike (P(NMDA spike)) in a terminal branch versus number of nearly synchronous stimulus-evoked synapses for peak synaptic NMDAR conductances of 1.0 nS (blue) and 0.5 nS (red) with and without background activity from 1500 excitatory synapses. Because of the higher AMPA/NMDA ratio, NMDAR spikes were identified by eye from their decay time (>15 ms) rather than peak depolarization for this simulation. (B) P(NMDA spike) versus number of nearly synchronous stimulus-evoked synapses for quantal NMDAR conductance amplitudes of 1.0 nS (blue) to 1.5 nS (red) with and without background activity of 900 excitatory synapses (reduced because increased gNMDA triggered NMDAR spikes with background activity from 1500 excitatory synapses). (C) P(NMDA spike) versus number of nearly synchronous stimulus-evoked synapses for NMDAR decay time constants of 70 ms and 35 ms (red) with and without background activity from 1500 excitatory synapses. Note: to compensate for the faster decay kinetics the background input rate was doubled (1.7 Hz) in order to deliver the same time averaged NMDAR conductance and depolarization. (D) NMDAR spike decay time distributions (N\u200a=\u200a100 across randomly selected branches) during control (dashed lines) and during background excitatory activity of 1500 synapses (solid lines), in the original model and in the model with faster NMDAR kinetics . With a decay constant of 35 ms NMDAR spikes are significantly shorter, both during control and during background excitatory activity. However, the increase over control produced by background was significant .(TIF)Click here for additional data file.Figure S5+, Ca2+ and Ih conductances on changes in NMDAR spike threshold and duration during background excitatory activity.Effect of changing dendritic Na (A1) Average probability of triggering an NMDAR spike (P(NMDA spike)) versus number of nearly synchronous stimulus-evoked synapses in the original model (black) and in a model where Na+ channels were removed from the apical tree (red) measured in the absence and presence of background activity from 1500 excitatory synapses . (A2) NMDAR spike decay time distribution for conditions in (A1). (B1) Average probability of triggering an NMDAR spike (P(NMDA spike)) versus number of nearly synchronous stimulus-evoked synapses in the original model (black) and in a model where L-type Ca2+ channels (Ca-L) were removed from the apical tree including Ca2+ initiation zone, during control (black) and background excitatory activity (B2) NMDAR spike decay time distribution for conditions in (B1). (C1) Average probability of triggering an NMDAR spike (P(NMDA spike)) versus number of nearly synchronous stimulus-evoked synapses in the original model (black) and in a model where Ih current was exponentially increased from 0 to 40 pS/\u00b5m2 along the apical tuft (red) measured in the absence and presence of 1500 background excitatory inputs . (C2) NMDAR spike decay time distribution for conditions in (C1).(TIF)Click here for additional data file.Figure S6+ conductances on changes in NMDAR spike threshold and duration during background excitatory activity.Effect of increasing dendritic K (A1) Average probability of triggering an NMDAR spike (P(NMDA spike)) versus number of nearly synchronous stimulus-evoked synapses in the original model (black) and in a model where K+ A-type (KA) channels density was doubled (60 pS/\u00b5m2) in the apical tree (red), measured in the absence and presence of 1500 background excitatory input . (A2) NMDAR spike decay time distribution for conditions in (A1). (B1) Average probability of triggering an NMDAR spike (P(NMDA spike)) versus number of nearly synchronous stimulus-evoked synapses in the original model (black) and in a model where K+ delayed rectifier (Kdr) channels density was increased ten-fold (10 pS/\u00b5m2) in the apical tree (red), during control and background excitatory activity . (B2) NMDAR spike decay time distribution for conditions in (B1).(TIF)Click here for additional data file.Figure S7+ conductances on the dendritic tuft to match levels reported by Harnett et al. 2013.Effect of increasing K (A) Average probability of triggering an NMDAR spike (P(NMDA spike)) versus number of nearly synchronous stimulus-evoked synapses in the original model (black) and in a model where apical KA density was increased to 77 pS/\u00b5m2 and apical Kdr density to 23 pS/\u00b5m2 to match estimates from B) NMDAR spike decay time distribution for conditions in (A). The decay time was computed as the time when the voltage decayed to 30% of the peak depolarization (instead of 37%) to account for the fluctuations present when the K+ conductance was increased (see G). (C) Average profile of NMDA spikes (N\u200a=\u200a100) triggered in the original model (grey) and in a model with K+ conductances equal to 1 and 2 times the levels reported by + conductance was required to completely counteract the background NMDAR component, matching the average NMDA spike recorded during background excitation with 6000 AMPAR-only synapse, which matched the depolarization during mixed AMPAR+NMDAR excitatory background (green). Note: the time course of this dendritic spike is markedly different from NMDA spikes recorded experimentally in L5 pyramidal cells D) Average number (N\u200a=\u200a28) of additional branches activated during background input \u00b1 SE when a single branch is nearly synchronously stimulated in the modified model (blue) compared to the original model (grey). (E) Example voltage traces from branch 12 (red) and branch 9 (blue) in the absence (dashed lines) and presence of 1500 background excitatory inputs (solid lines). (F) Peak depolarization induced by an NMDAR spike in branch 12 versus terminal branch number in absence and presence of background activity. (G) Local voltage profile of an NMDA spike triggered in the modified model by the near synchronous activation of 30 glutamatergic synapses. Current generated by each of the 30 NMDAR synapses compared with the current densities of the three major ions . Note: another recent study dr (3 pS/\u00b5m2) in the apical dendrites of L5 pyramidal cells.(TIF)Click here for additional data file.Figure S82+-dependent local feedback mechanisms on NMDAR spikes and spatio-temporal integration.Effect of Ca (A) Individual quantal synaptic NMDAR conductance with Ca2+-dependent inactivation (CDI) present in the original model (black trace) and when CDI was removed (red line), both recorded at \u221260 mV. (B) Ca2+ current through the NMDAR quantal conductance with (black trace) and without (red line) CDI at \u221260 mV, together with scaling factor for NMDA conductance (lower plot). (C) Branch voltage during a single NMDAR spike with (dashed traces) and without (solid traces) CDI of NMDARs, in the absence (red) and presence of background activity from 1500 excitatory synapses (blue). The effect of CDI became evident during the longer plateau potential, because Ca2+ accumulation lead to shortening of NMDAR conductances. (D) NMDAR spike decay time distribution for conditions in (C). During background excitatory input the average NMDAR spike duration increased 4.8 fold with background excitation without CDI, compared to 1.4 in the original model. (E) Average NMDAR spike decay time in the absence and presence of background excitatory input from 1500 excitatory synapses (solid marker), with different levels of negative feedback implemented with an SK-like Ca2+-dependent K+ conductance at each synapse (SKsyn). SKsyn at a density of 40 pS/synapse reproduced the average decay time in the original model (orange symbol). (F) Cumulative distributions of NMDAR spike decay times with different levels of SKsyn in the absence and presence of background excitation (solid markers). All levels tested show a significant increase during background excitation compared to control (Kolmogorov-Smirnov test: P<0.05). (G) Average probability of action potential (P(AP)) with different levels of negative feedback in the absence and presence of background excitatory input , for different numbers of distributed synapses stimulated with random trains at 10 Hz for 100 ms.(TIF)Click here for additional data file.Movie S1Movie 1 shows the membrane voltage in the Layer 5 Pyramidal cell model during nearly synchronous stimulus evoked synaptic stimulation with 30 synapses on a terminal dendritic branch (each synapse firing at 200 Hz from 100\u2013105 ms). The voltage response to the synaptic input is shown under 3 conditions: 1) Control with no background activity. 2) With background activity from 1500 excitatory synapses firing randomly at 0.85 Hz. 3) With balanced background synaptic input including inhibition from SOM-like (GABAA) and NGF-like (GABAA and GABAB) mediated inhibition. While the spread of depolarisation in the control case is limited to the stimulated branch and its near neighbours, the excitatory background input causes regenerative depolarisation in branches further away. The spread of depolarisation is also widespread for balanced background synaptic input, but the presence of inhibition shortens the duration compared with background excitation alone.(AVI)Click here for additional data file.Movie S2Movie 2 shows the membrane voltage in the Layer 5 Pyramidal cell model during nearly synchronous stimulus evoked synaptic stimulation of 100 synapses spatially distributed over the apical tuft (10 Hz from 100\u2013200 ms) under 3 conditions: 1) Control with no background activity. 2) With background activity from 1500 excitatory synapses firing randomly at 0.85 Hz3) With balanced background synaptic input including inhibition from SOM-like (GABAA) and NGF-like (GABAA and GABAB) mediated inhibition. Only in the case of stimulation in the presence of excitatory input did the spatially distributed synaptic input trigger somatic action potentials which back propagates into the apical tree.(AVI)Click here for additional data file.Text S1Supporting information. Supplemental methods, results, table and references.(DOCX)Click here for additional data file."} +{"text": "Quinpramine exerted its anti-proliferatory effect on antigen presenting cells, but not on responder T cells. Our data suggest that quinpramine represents a candidate pharmaceutical for inflammatory neuropathies.Activation of inflammatory cells is central to the pathogenesis of autoimmune demyelinating diseases of the peripheral nervous system. The novel chimeric compound quinpramine\u2014generated from imipramine and quinacrine\u2014redistributes cholesterol rich membrane domains to intracellular compartments. We studied the immunological and clinical effects of quinpramine in myelin homogenate induced Lewis rat experimental autoimmune neuritis (EAN), a model system for acute human inflammatory neuropathies, such as the Guillain-Barr\u00e9 syndrome. EAN animals develop paresis of all limbs due to autoimmune inflammation of peripheral nerves. Quinpramine treatment ameliorated clinical disease severity of EAN and infiltration of macrophages into peripheral nerves. It reduced expression of MHC class II molecules on antigen presenting cells and antigen specific T cell proliferation both Acute inflammatory neuropathies are rare The prototypic acute inflammatory neuropathy Guillain\u2013Barr\u00e9 syndrome (GBS) most commonly manifests as ascending flaccid tetraparesis with less pronounced sensory deficits in vitro screen for compounds inhibiting prion protein amplification the chimeric drug quinpramine \u2013 fused from the anti-depressant imipramine and the malaria treatment quinacrine \u2013 was highly effective A promising novel compound for the possible future treatment of inflammatory disorders of the nervous system has recently been identified. In an In experimental autoimmune encephalomyelitis (EAE) \u2013 the animal model of multiple sclerosis \u2013 quinpramine reduced clinical disease severity in both a preventive and therapeutic paradigm EAN was induced as previously described in vitro application, quinpramine was dissolved in DMSO (1 mg/ml) and used at a final concentration of 50\u2013200 nM for 1\u201324 hours in quadruplicate wells.Quinpramine was synthesized as previously described 2 narcosis and sciatic nerves were dissected and paraffin or epoxy resin embedded as previously described At the peak of EAN severity (day 14 post immunization) a randomly chosen half of the treated group (n\u200a=\u200a5) was sacrificed by COThe murine macrophage cell line RAW264.7 (ATCC) was cultured in the presence of quinpramine (200 nM) for six days before staining for cholesterol with filipin in 90% glycerol and 1.5 mg/ml glycin in PBS for 30 minutes at room temperature. Stained cells were photographed using a conventional microscope .5/well) were cultured in flat bottom 96-well plates in standard T cell medium in the presence of varying concentrations of BPNM (0.1\u2013100 \u00b5g/ml) in quadruplicate wells for 96 hours. 3H-Thymidin was added for the last 24 hours and cell proliferation was measured by detection of incorporated radioactivity as counts per minute.At day 14 post immunization spleens from randomly chosen quinpramine and vehicle treated EAN rats were dissected under sterile conditions and passed through a 40 \u00b5m cell strainer followed by ammonium chloride based erythrocyte lysis (BD Bioscience). Derived responder splenocytes (2\u00d710323\u2013339) (OTII cells) . OTII cells were cryopreserved in 50% DMEM, 40% FCS and 10% DMSO and thawed immediately before usage. Cocultures were prepared using responder OTII cells and irradiated (1000 Gy) syngenic antigen presenting cells (APC) in the presence of Ova323\u2013339 and proliferation was measured by 3H-Thymidin incorporation. Either APCs or responder cells were pre-incubated before coculturing with quinpramine at varying dosages (0\u2013200 nM) for 12 hours. Combinations of differently pre-treated cells were tested for proliferation in quadruplicate wells and two independent experiments were performed.Mouse splenocytes were prepared from OTII mice expressing a T cell receptor recognizing the MHC- II restricted Ovalbumin peptide amino acids 323\u2013339 -\u03b3 in the presence or absence of quinpramine (100 nM). The percentage of MHC-II+ cells and the fluorescence intensity of MHC-II staining were assessed. Three independent experiments with quadruplicate wells were performed. To assess the fluorescence pattern of quinpramine and its precursor substances, splenocytes were cultured under routine conditions for 1\u20136 hours in the presence of quinpramine (100 nM), quinacrine (100 nM), imipramine (100 nM) or a mixture of the later two (each at 100 nM). Cells were washed three times with PBS and analyzed for fluorescence in the channels FITC, PE, PerCP-Cy5.5, PE-Cy7, APC, APC-Cy7, PacBlue, AmCyan. All flow cytometry was performed using a FACSCanto II flow cytometer. Two independent experiments with triplicate wells were performed. All flow cytometry data were analyzed using FlowJo software (v7.2.5 TreeStar).For extracellular staining, splenocytes from quinpramine or vehicle treated rats were co-stained for cell surface CD11b/c (BD 554862) and MHC class II using a DyLight649 labelled goat-anti-mouse secondary antibody (Serotec STAR74D649). For intracellular staining cells were first stained with CD11b/c, fixed with 4% paraformaldehyde solution, permeabilized with PermBuffer and subsequently stained for intracellular MHC-II according to the manufacturers protocol (BD Biosciences). To analyze the MHC-II distribution Data were statistically analyzed using GraphPadPrism 5.0 (GraphPad Software). The Wilcoxon-Mann-Whitney was used to test for statistically significant differences in clinical score values. Student's t-test for unrelated samples was used to test for statistically significant differences in all other analyses. Differences were considered significant at p-values<0.05.quinacrine did not ameliorate clinical signs of EAN and whens of EAN , suggestar cells . We founar cells . The nerar cells . The denar cells and of Mar cells cells waar cells . This inex vivo isolated splenocytes between the three cohorts. Splenocytes from quinpramine treated animals had lower percentages of MHC-II+ cells in total and of CD11b/c+ APCs were sufficient to reduce peak EAN severity on day 14 suggestiGiven that numerous compounds are effective in EAN, but none of these has reached the bench-to-bedside transfer In conclusion, our data suggest that quinpramine represents a possible candidate pharmaceutical to further establish in acute inflammatory neuropathies and that quinpramine takes effect on MHC class II expression on APCs. Such a mechanism may also be effective in other autoimmune disorders and may extend our therapeutic armentarium in autoimmune mediated disorders.Figure S1Quinacrine treatment does not ameliorate EAN. Moderate EAN was induced in female Lewis rats (n\u200a=\u200a6 per group), who received oral vehicle or quinacrine (2 mg/kg/week) treatment \u2013 one of the precursor substances of quinpramine \u2013 starting at day 2 after immunization. Quinacrine treatment (green inverted triangles) did not significantly alter severity or course of moderate EAN in comparison to vehicle treatment (black points). Plots represent mean \u00b1 SEM.(TIF)Click here for additional data file.Figure S2Quinpramine treated cells exhibit a specific pattern of fluorescence. (A) Cultured rat splenocytes were left untreated (red line) or incubated for six hours with quinpramine or a mixture of quinacrine and imipramine and analyzed by multicolor flow cytometry. Flow cytometry histograms depict that both quinpramine and mixture treated cells returned green fluorescence in the FITC (left panel) and AmCyan (middle panel) channel. (B) Average fluorescence intensities were calculated for each channel and quinpramine (green bars), mixture of quinacrine and imipramine (blue bars) and quinacrine only (yellow bars) all returned specific patterns of fluorescence in the FITC (left panel) and AmCyan (middle panel) channel. Imipramine (white bars) did not generate fluorescence in comparison to untreated controls (red bars). Notably, a mixture of quinacrine and imipramine did not return the same fluorescence intensity as quinpramine alone indicating that quinpramine is not cleaved to its precursors within the cells.(TIF)Click here for additional data file."} +{"text": "To assess if flow-sensitive four-dimensional velocity encoded cine magnetic resonance imaging (4D VEC MRI) adds value in diagnosing patients with suspected partial anomalous pulmonary venous drainage (PAPVD).In six patients with echocardiographically suspected PAPVD, anatomy was evaluated using standard magnetic resonance imaging including angiography. Functional analysis included shunt calculations from standard flow-measurements. Furthermore, 4D VEC MRI was used for visualization of maldraining pulmonary veins and quantification of flow via the maldraining veins and interatrial communications, if present.In all patients, the diagnosis of PAPVD was confirmed by standard magnetic resonance imaging. Shunt volumes ranged from 1.4:1 to 4.7:1. Drainage sites were the superior caval vein (n=5) or the vertical vein (n=1). Multiple maldraining pulmonary veins were found in three patients. Pulmonary arteries and veins could be clearly distinguished by selective visualization using 4D VEC MRI. Flow measured individually in maldraining pulmonary veins (n=6 patients) and across the interatrial communication (n=3 patients) revealed a percentage of the overall shunt volume of 30 to 100% and 58 to 70%, respectively. Different flow characteristics and directions of maldraining pulmonary veins and across the interatrial communication could be visualized during ventricular systole and diastole.Selective visualization of individual vessels and their flow characteristics by 4D VEC MRI facilitates to distinguish adjacent pulmonary arteries and veins and thus improves accurate diagnosis of maldraining pulmonary veins. By detailed quantification of shunt volumes additional information for planning of treatment strategies is provided. This method adds clinical value and might replace contrast-enhanced magnetic resonance angiography in these patients in the future.Selective visualization of streamlines in a child diagnosed with anomalous pulmonary venous drainage of the right upper pulmonary vein into the superior caval vein (SCV) and presence of an interatrial communication in different perspectives.Red streamlines represent the flow of the maldraining right upper pulmonary vein (RUPV) entering the right atrium (RA) together with the blue streamlines of the SCV (panel A and B).The green streamlines in the pulmonary trunk (PT), right and left pulmonary artery (RPA and LPA) allow distinguishing clearly peripheral pulmonary arteries (green) and veins .The yellow streamlines of the anatomic correctly draining right lower pulmonary vein (RLPV) merge with the red streamlines representing a left to right shunt across the interatrial communication and thus functional maldrainage during ventricular systole (panel D)."} +{"text": "Postoperative ventricular dysfunction (VnD) occurs in 9\u201320% of coronary artery bypass graft (CABG) surgical patients and is associated with increased postoperative morbidity and mortality. Understanding genetic causes of postoperative VnD should enhance patient risk stratification and improve treatment and prevention strategies. We aimed to determine if genetic variants associate with occurrence of in-hospital VnD after CABG surgery.A genome-wide association study identified single nucleotide polymorphisms (SNPs) associated with postoperative VnD in male subjects of European ancestry undergoing isolated primary CABG surgery with cardiopulmonary bypass. VnD was defined as the need for \u22652 inotropes or mechanical ventricular support after CABG surgery. Validated SNPs were assessed further in two replication CABG cohorts and meta-analysis was performed.\u22124), with one SNP (rs17691914) encoded at 3p22.3 reaching genome-wide significance (Padditive model\u200a=\u200a2.14\u00d710\u22128). Meta-analysis of validation and replication study data for 17 SNPs identified three SNPs associated with increased risk for developing postoperative VnD after adjusting for clinical risk factors. These SNPs are located at 3p22.3 , 3p14.2 and 11q23.2 .Over 100 SNPs were associated with VnD (P<10additive model>2.1) of developing in-hospital VnD after CABG surgery. However, three genetic loci identified by meta-analysis were more modestly associated with development of postoperative VnD. Studies of larger cohorts to assess these loci as well as to define other genetic mechanisms and related biology that link genetic variants to postoperative ventricular dysfunction are warranted.No SNPs were consistently associated with strong risk (OR Each year almost 250,000 patients undergo coronary artery bypass graft (CABG) surgery in the United States alone, with the primary aim of surgery being to prevent major adverse cardiovascular events NPPA/NPPB and NPR3 natriuretic peptide system genes that associate with VnD after CABG surgery with adjusted odds ratios for association >2.0 We have previously examined candidate genetic loci and have identified single nucleotide polymorphisms (SNPs) within the Ethics Statement: In compliance with the Declaration of Helsinki, respective Institutional Review Board approval and subject written informed consent were obtained for all subjects .Data were collected prospectively from primary CABG surgical patients at three institutions: Brigham and Women's Hospital , the Texas Heart Institute , and Vanderbilt University Medical Center . These institutions enrolled subjects into two cardiac surgical cohorts: the CABG Genomics Program (BWH and THI) and the Vanderbilt Cardiac Surgery Registry (VCSR). De-identified control genotypes were provided for the GWAS from an ambulatory multiple sclerosis study cohort of European ancestry subjects. CABG subjects were prospectively excluded from analyses if they had emergency surgery, re-operative cardiac surgery, concurrent valve surgery, no aortic cross-clamp, or CPB >220 minutes, or if preoperatively they had moderate to severe mitral regurgitation, LVEF <20%, severe preoperative renal dysfunction , or were receiving inotrope, intra-aortic balloon pump or ventricular assist device support. To minimize false positive SNP associations related to population stratification, all analyses were restricted to European ancestry subjects. With the exception of the GWAS' controls, analyses did not include women, as gender may influence development of heart failure, and the number of female subjects with VnD enrolled in the CABG Genomics and VCSR cohorts (<20%) was insufficient to support gender stratified SNP association analyses Postoperative VnD cases were identified in accordance with prior studies \u22126). SNPs were also excluded if they had a plate association (P<10\u221210) or differential missingness either between cases and controls (P<0.001) or based on flanking haplotypes .Genotypes were ascertained using the Affymetrix 6.0 Genome-Wide Human SNP Array and were analyzed using the Birdseed (version 2) algorithm. SNPs were excluded from analyses for call rates <95%, minor allele frequencies <1%, or for not exhibiting Hardy Weinberg equilibrium and the manufacturer's protocols. Sequenom raw data were analyzed with the SpectroTyper 3.4 software . Spectra and cluster plots were checked by visual inspection of intensity plots with manual curation of genotype calls.Subjects were excluded from GWAS analyses if they did not self-identify to be of European ancestry. In addition, population stratification outliers were excluded from GWAS analyses as described in Because the cohort size of this exploratory study would detect only genetic variants with strong associations with development of postoperative VnD, a Type 1 error rate <0.0005 was utilized for statistical power assessments. This P value threshold was chosen with the understanding that a proportion of SNP associations detected at this level of significance could be false positive associations and that further assessment would be conducted beyond the initial GWAS. Based on the available case to control ratio of 1\u223614 and assuming an additive genetic model, 12% incidence of VnD, 80% power, minimum estimated detectable genetic risk ratios for the GWAS were 2.1, 2.2, 2.3, 2.5, and 2.8 for SNPs with minor allele frequencies of >0.25, 0.25, 0.20, 0.15, and 0.10 respectively Association between SNPs and VnD were estimated using the trend test for the additive genetic model, the Pearson chi-square test for allelic and dominant genetic models, and the Fisher's Exact test for the recessive genetic model. Using genomic inflation factor lambdas genomic-control corrections were made to account for residual population stratification effects suggested by Q-Q plots . ClusterCategorical and continuous patient demographic and clinical characteristics were compared between case and control groups for the CABG Genomics validation study, Vanderbilt replication study, and CABG Genomics replication study using Pearson's chi-squared, Fisher's Exact or student's t-tests where appropriate . PLINK (version 1.07) was used for genetic analyses Univariate SNP associations with VnD were assessed using Pearson chi-square tests for the allelic model and logistic regression for the additive, dominant and recessive models. SNP associations were additionally adjusted for age, LVEF, CPB time, and institution using logistic regression. Univariate and multivariable adjusted odds ratios (OR) and 95% confidence intervals (CI) were determined for each SNP. Point-wise permutation analyses were conducted for each SNP (univariate and multivariable covariate adjusted assessments). In order to assess that observed SNP associations with VnD were not a consequence of associations with presenting LVEF, linear regression was used to conduct univariate assessments of CABG Genomics validation study SNPs for association with preoperative LVEF. Meta-analyses were conducted for SNPs successfully genotyped for both replication studies . All VnD cases had significantly lower preoperative LVEFs and longer CPB times than CABG controls (P<0.05), and CABG Genomics cases used for validation had more subjects with a >30 pack year history of smoking than CABG controls (P<0.05). VnD cases assessed in validation and replication studies differed significantly between groups with regards to preoperative LVEF, but not other characteristics.We surveyed the genomes of 92 male VnD cases using Affymetrix 6.0 arrays. Sixteen samples were subsequently excluded from analysis based upon genotyping quality control, gender mismatch, population stratification outliers, and cryptic relatedness (methods described in \u22124 (\u22128) and additive models (P\u200a=\u200a2.1\u00d710\u22128). This SNP had minor allele frequencies of 21% and 7% in VnD cases versus controls, respectively. An additional 10 SNPs associated with VnD with P values <10\u22125. Three of these 10 SNPs reside within the same chromosome 3 locus as rs17691914 . Of the SNPs with P values <10\u22125, rs2171325 was not assessed in further validation or replication studies because the minor allele frequency was <10% in both cases and controls. Analyses of 709,355 SNPs identified 123 SNPs (63 genetic loci) that were associated with VnD with P values <10\u22124 . One SNP\u22124 for association with VnD were selected for further assessment in CABG Genomics cases and controls. Five samples that failed \u226590% of validation study genotyping and one SNP (rs11060480) that did not technically validate were excluded from further study. After clinical and genotyping exclusions, data for 71 of the 76 GWAS VnD cases and 715 male CABG controls (subjects without VnD) were analyzed. The technical validation aspect of this study compared minor allele frequencies for the VnD cases for Affymetrix 6.0 versus Sequenom genotyping in order to identify potential false positive GWAS associations due to technical calling errors. In addition, this study refined the initial genetic association study in order to: 1) assess if GWAS associations persisted when the VnD case data was compared to male CABG control data; 2) allow assessment of additional SNPs with moderate linkage disequilibrium with identified GWAS SNPs (correlation r2 with GWAS SNPs\u200a=\u200a0.30\u20130.70) in order to identify SNPs within GWAS loci with potentially stronger associations with VnD; 3) allow covariate adjustment of SNP associations for age, institution, LVEF and duration of CPB; 4) allow assessment of SNP associations with preoperative LVEF.GWAS SNPs with a minor allele frequency>10% and P<10Sixty-three SNPs (36 GWAS SNPs and 27 SNPs in linkage disequilibrium with these SNPs) were assessed in 71 VnD cases and 715 CABG controls . Twenty-additive model\u200a=\u200a1.65, P\u200a=\u200a0.04; covariate adjusted ORadditive model\u200a=\u200a1.69, P\u200a=\u200a0.056). This association is not significant after Bonferroni adjustment.The 19 SNPs selected for replication were assessed initially in the VCSR cohort. Two SNPs, rs9837024 and rs10773689, were excluded due to failed genotyping and Hardy Weinberg Equilibrium testing in controls, respectively. Results for the remaining 17 SNPs (encoded at 13 loci) are shown in We also assessed replication in the CABG Genomics cohort using previously unstudied cases. All 19 SNPs were successfully genotyped, and for 12 SNPs (encoded at 10 loci), the direction of the ORs was consistent across the GWAS and validation studies. However, none of these 12 SNPs achieved significant association with VnD (P<0.05) in univariate or multivariable adjusted analyses .2 heterogeneity index\u200a=\u200a0). However, this association does not achieve statistical significance after Bonferroni adjustment.To assess for more modest risk (OR <2.1) that might be associated with putative VnD SNPs, we performed a meta-analysis of the VCSR and CABG Genomics cohort replication studies. SNP rs17061085 was associated with postoperative VnD with the same direction of effect as the GWAS, even after adjusting for age, preoperative LVEF and CPB time and non-VnD CABG control (n\u200a=\u200a1200) data from the validation and replication studies using meta-analysis . Table 3VnD after cardiac surgery is associated with significant patient morbidity and mortality. Although clinical parameters such as preoperative LVEF and duration of CPB are predictive of risk for developing postoperative VnD PDCD6IP). Despite a paucity of other genes within close proximity to rs17691914, we are intrigued that this SNP achieved genome-wide significance in the initial GWAS and that the 3p22.3 is also tagged by a SNP (rs12638540) within an intron of the CKLF-like MARVEL transmembrane domain containing 7 gene (CMTM7) that is reported to be significantly associated with heart failure mortality in a GWAS meta-analysis of 2,526 ambulatory patients Future study of the genes at the 3 loci tagged by these VnD-associated SNPs may enhance understanding of the pathophysiology of postoperative VnD. The 3p22.3 locus (defined by rs17691914) is located within 1 Mb of the programmed cell death 6-interacting protein gene (FHIT) gene that encodes a protein implicated in multiple cancers FHIT has not been well studied in cardiac tissue, human atrial expression of FHIT is diminished in the setting of decreased LVEF SNP rs17061085, which maps to 3p14.2, resides near the fragile histidine triad gene that encodes a protein implicated in facilitating the invasion, migration and metastasis of human tumor cells via epithelial-mesenchymal transitions IL10RA), which may participate in the oxidative-stress responses that promote cardiac remodeling and heart failure SCN4B and SCN2B) that are expressed in cardiac myocytes and are mutated in long-QT syndrome and atrial fibrillation, respectively The 11p11.2 intronic SNP rs12279572 lies within the We recognize several important limitations of our study, including the small size of the study cohorts and the narrow demographic profiles we were consequently able to examine. We consider this study to be an exploratory assessment for strong genetic influences on the postoperative ventricular dysfunction outcome, as cohort size restricted our ability to detect SNPs associated with VnD with genetic risk ratios <2.1 for an additive genetic model. We secondarily assessed dominant and recessive genetic models, albeit with statistical power to detect substantially higher genetic risk ratios for these models than for the additive model. Our findings have important implications for design of future studies, in that no SNPs consistently associated with postoperative VnD with additive model ORs >2.1. However, GWASs of cardiovascular disease in very large non-surgical populations illustrate the potential to discover SNPs associated with more modest risks of developing common disease. For example, loci found to be associated with coronary artery disease and/or myocardial infarction NPPA/NPPB gene complex on chromosome 1 that significantly associate with VnD after CABG surgery (additive model ORs 1.85\u20132.29)NPPA/NPPB SNPs were not assessed in the Affymetrix 6.0 SNP array platform. However, the Affymetrix 6.0 Genome-Wide array is estimated to cover \u223c83% of genome-wide SNP variability, thus providing very good potential for this study to detect novel SNP associations with VnD after CABG surgery We recognize that gaps in the Affymetrix 6.0 Genome-Wide array may still potentially miss important SNP associations with VnD, as all commercially available genome-wide SNP arrays have some gaps in coverage of genetic variability across the genome Another important limitation of this study was the narrow demographic profile of the analyzed study cohorts. There were not enough women enrolled within the CABG Genomics and VCSR cohorts to allow for SNP association analyses stratified by gender. Nor were there sufficient non-European ancestry subjects of either gender to allow for SNP association analyses stratified by ethnicity. Limiting this study to men of European ancestry enhanced homogeneity of the study cohort for the purpose of best allowing detection of real SNP associations with VnD that were not confounded by influence of gender and racial background. However, we therefore propose that further evaluations of VnD-associated SNPs be conducted in women and in both men and women of non-European ancestry when cohorts are available that contain sufficient sample sizes of these populations.In conclusion, at least 3 loci identified by genome-wide and subsequent validation and replication assessments were associated with modest risk for VnD after primary CABG surgery. These and other SNPs within the surrounding genetic loci warrant further replication in large cardiac surgical cohorts. With continued identification of genotypes associated with postoperative VnD, we expect to improve risk stratification and gain insights into biologic pathways linking these variants to postoperative myocardial dysfunction. Such knowledge could lead to new preventions and therapies for postoperative VnD, and, possibly other forms of heart failure.Figure S1Genome wide association study Q-Q plots of the expected and observed \u2013log P values for SNP associations with ventricular dysfunction after primary coronary artery bypass graft surgery. Genomic inflation factor lambdas derived from these Q-Q plots were used to adjust GWAS association findings for potential population stratification: allelic model lambda\u200a=\u200a1.08, additive model (trend test) lambda\u200a=\u200a1.07, dominant model lambda\u200a=\u200a1.05, and recessive model lambda\u200a=\u200a1.00.(TIF)Click here for additional data file.Figure S2Examples of single nucleotide polymorphism (SNP) genotyping cluster plots with good and poor differentiation between heterozygotes and homozygotes. SNP\u200a=\u200asingle nucleotide polymorphism; NC\u200a=\u200ano genotype call; AA\u200a=\u200ahomozygote genotype for one allele; AB\u200a=\u200aheterozygote genotype call; BB homozygote genotype call for other allele Two investigators visually inspected intensity cluster plots for genotyping calls for those SNPs with GC adjusted association P values<1.0\u00d710\u22124 in the GWAS . Intensity cluster plots for recessive model SNPs with association GC adjusted P values<10\u22125 in the GWAS as determined by chi-square tests were also reviewed, and P values for SNPs with good cluster plot differentiation were then assessed using Fisher's exact tests Click here for additional data file.Information S1Supplementary Methods.(DOC)Click here for additional data file.Table S1Genome wide association study results: SNP associations with ventricular dysfunction after primary coronary artery bypass graft surgery.(DOC)Click here for additional data file.Table S2Validation study results: SNP associations with ventricular dysfunction after primary coronary artery bypass graft surgery in 786 European ancestry men from the CABG Genomics Study Cohort. 63 SNPs were selected from 34 genetic loci identified to be associated with ventricular dysfunction in the GWAS.(PDF)Click here for additional data file.Table S3Vanderbilt cohort replication study results: 17 SNP associations (13 genetic loci) with ventricular dysfunction after primary coronary artery bypass graft surgery in 337 European ancestry men.(DOC)Click here for additional data file.Table S4CABG Genomics cohort replication study results: 19 SNP associations (15 genetic loci) with ventricular dysfunction after primary coronary artery bypass graft surgery in 980 European ancestry men.(DOC)Click here for additional data file.Table S5Covariate adjusted meta-analysis results: 17 SNP associations (13 genetic loci) with ventricular dysfunction after primary coronary artery bypass graft surgery in 1388 European ancestry men.(DOC)Click here for additional data file."} +{"text": "Women with genetic cardiovascular diseases,or repaired congenital defects require additional monitoring during pregancy and may need planned surgical delivery of the infant. CMR is a simple, accurate multi parametric imaging techniques which provides assessment of cardiac anatomy, function and flow in the pregnant patient.A retrospective analysis of the cardiac and obstetric notes of 5 women was carried out to evaluate the use of CMR, the need for further imaging/intervention and outcome of the pregnancy.5 women aged 22-30 (mean 26.6 years) were referred for CMR during 1st (3) pregnancy and 2nd pregnancy(2). Patients were 16-28 weeks pregnant(mean 22 weeks) at the time of imaging Previous medical history and indications for CMR were: severe aortic stenosis and bicuspid valve with syncopal episodes (1), Marfans syndrome with chest pain, aortic dissection on trans oesophageal echo (1), aortic coarctation and increasing aortic root dimensions on trans thoracic echocardiography (1), aortic coarctation and bicuspid aortic valve, patent ductus arteriosus and ventricular septal defect presenting with chest pain (1) and investigation of a cardiac source of emboli causing ischaemic episodes in right leg (1).Imaging was carried out on a 1.5T Siemens Sonata or Avanto using a spine array posteriorly and 2 channel flex array placed over the anterior chest. Patients were placed supine then a wedge was placed under the right hip to minimise aorto-caval compression. All images were acquired using ECG gating and sequences limited to answer the clinical question. Acquisitions were kept at normal operating mode with an specific absorption rate (SAR) of 2w/kg. Imaging protocol comprised SSFP (steady state free precession) localisers in three orthogonal planes; anatomical assessment in axial, coronal and sagittal/sagittal planes using HASTE long and short axis SSFP retrospectively gated breathold cines for the visual assessment of ventricular function. Short axis images were acquired for accurate analysis of ventricular function and mass . Phase shift velocity mapping was added for the evaluation of flow and peak velocity in patients with valvular disease or aortic coarction.All patients tolerated CMR- average imaging time of 25mins. 4 patients delivered at 34-37 week (mean 35 weeks) 3 planned caesarean sections,1 spontaneous vaginal delivery.Birth weight 2370-3076 grams (mean 2589 grams).Balloon valvuloplasty was carried out within 1 week of CMR in the patient with severe aortic stenosis and surgical aortic valve replacement 3 months post delivery. No further CMR has been performed. No evidence of dissection found in patient with Marfans syndrome but aortic root was dilated(4.1 cm) which remained stable 2 months post delivery and 3 years later. No cardiac cause of emboli was found in the patient with right leg ischaemic episodes and no cause for chest pain or recurrence of re-coarctation seen in the remaining patients and no further CMR has been performed.CMR is a safe reliable imaging method for the diagnostic assessment of the aorta and cardiac function in pregnancy It can be used as a screening tool pre, during and post pregnancy and can guide intervention and aid planning delivery.Jane Francis is funded by OCMR- Oxford Centre for Clinical Magnetic Resonance Research, University of Oxford."} +{"text": "Plasmodium falciparum isolates.Rapid diagnostic tests (RDTs) have revolutionized the diagnosis of malaria. Among the various factors affecting RDTs sensitivity is genetic variation of the antigen used. The genetic variation in PfHRP2 and PfHRP3 proteins was studied among the Indian P. falciparum were collected from six geographical regions of India. Target genes encoding PfHRP2 and PfHRP3 antigens were sequenced to study genetic polymorphism. Minimum detection limit giving a positive rapid diagnostic test was also determined.One hundred and forty isolates of Extensive variations were observed in amino acid repeat types of PfHRP2 and PfHRP3. PfHRP2 exhibited more polymorphism than PfHRP3. Significant relation was observed between type 2 and type 7 repeats and RDT detection rate as higher number of these repeats showed better sensitivity with RDTs.Pfhrp2 and Pfhrp3 genes among Indian P. falciparum population and its relation to RDT sensitivity.The results provide insights into the genetic diversity of Plasmodium falciparum-specific histidine rich protein 2 (PfHRP2). Other RDTs use lactate dehydrogenase and aldolase and can also detect non-falciparum infections. Most of the RDTs show excellent sensitivity and specificity for P. falciparum at a parasitaemia greater than 500 parasites per microlitre (parasites/\u03bcl) but at lower parasitaemia, the sensitivity is variable for P. falciparum hrp3 sequences.The nucleotide sequences reported here have been deposited in the GenBank database under accession numbers for RDTs: Rapid Diagnostic Tests; HRP-2: Histidine Rich Proteins-2; PCR: Polymerase Chain Reaction; WHO: World Health Organization; ACT: Artemisinin based Combination Therapy; CQ: Chloroquine; NVBDCP: National Vector Borne Disease Control Programme; SPSS: Statistical Package of Social Services; CHC: Community Health Centre; PHC: Primary Health Centre.Authors declare that they have no any competing interests.NK, JPNS, RK, BS performed experimental work, data analysis and manuscript writing. VP and NM gave constructive advice and review the manuscript. NV and AA were involved in all stages of this study. All authors have read and approved the final version of manuscript.Table S1. Minimum parasite densities among P. falciparum isolates/lines from different regions of India.Click here for file"} +{"text": "To the Editor: To date, the genus Kobuvirus has consisted of 2 officially recognized species, Aichi virus and Bovine Kobuvirus of 60 stool samples collected from pigs in Hungary were positive for porcine kobuvirus by reverse transcription\u2013PCR (RT-PCR) . Phylogenetic and molecular evolutionary analyses were conducted by using MEGA 4 . Thirty representative strains of porcine kobuvirus detected in this study were randomly selected, sequenced, and analyzed to determine their evolutionary relationships with other kobuvirus reference strains. The partial 3D region among all 30 porcine kobuvirus strains was highly conserved, with nucleotide sequence identities >90%. In addition, our strains were most closely related to 2 porcine kobuvirus reference strains (S-1-HUN and Swine/2007/CHN) available in GenBank, with the nucleotide sequence identity ranging from 91.5% to 96.3%. Phylogenetic analysis of partial 3D nucleotide sequences of our porcine kobuvirus strains, together with published sequences of porcine kobuvirus reference strains (and those of Aichi virus and bovine kobuvirus), is shown in the Porcine kobuviruses have previously been reported only in healthy pigs ("} +{"text": "Identification of genetic variations in genes related to metabolism of selenium, including glutathione peroxidases (GPX) and thioredoxin reductases (TXNRD) as markers for breast cancer and/or ovarian cancer risks in carriers of BRCA1 gene mutation: analysis of selected changes in the subgroups, depending on the plasma selenium concentration.A group of 50 newly diagnosed breast and/or ovarian cancer patients carrying BRCA1 mutation was analyzed by exon-by-exon sequencing of 3 genes coding selenoproteins. Simultaneously a nested case-control study of 39 women with breast cancer and 7 women with ovarian cancer and 92 controls matched 1 to 2 cases has been conducted and changes detected by sequencing have been analyzed. Additional case \u2013 control studies were performed on 45 pairs 1:2 matched for GPX1 (rs1050450). All cases and controls were matched for age at enrolment, past history of breast cancer and oophorectomy. All these patients were carriers of one of three Polish founder BRCA1 mutation. In these patients plasma selenium level has been determined.The following techniques for laboratory analyses have been applied: a) sequencing on (ABI310), b) SimpleProbe or Taqmqan analysis (a melting-curve genotyping with fluorescence-labeled probes based on the LightCycler 480 System (Roche Applied Science)), c) determination of selenium concentration in plasma using atomic absorption spectrometer AAnalyst600 (Perkin Elmer).The strongest association has been found for GPX1 (rs1050450). Multivariate statistics allowed the analysis of each risk factor separately as well as their interaction with the help of logistic regression model. Subjects having plasma selenium levels under or equal 80 \u03bcg/l showed a generally decreased risk of being diseased . The genotype of rs1050450 in the gene GPX1 was not a risk factor for itself (p=0.70). However, when analyzed in combination, an interaction effect could be proven where carriers of the minor allele of rs1050450 having plasma selenium levels above or equal 80\u00b5g/l were under an increased disease risk .This effect was even more exacerbated among those women which joined the study after adnexectomy , while among women without adnexectomy the effect could not be detected at all . Adnexectomy was not a risk factor itself in the logistic regression model (p=0.49). Cases and controls in triplets with the same genotypes of GPX1 were also taken into account. Although no significant result was achieved, there was a tendency for more unaffected subjects of CC genotype-carriers in GPX1 with serum selenium levels above or equal 80\u03bcg/l (5:14 instead of expected 5:10). The opposite was shown for non-CC genotype carriers in GPX1 with plasma selenium level above or equal 80\u03bcg/l (12:18 instead of expected 12:24)."} +{"text": "Stress perfusion Cardiac MR is increasingly used in patients with established coronary artery disease (CAD) to guide further management. Our tertiary cardiac centre caters to a population of 1.8 million, performing over 4500 coronary angiograms and 1500 adenosine stress CMR scans/year.Our aim was to audit our clinical practice to determine the influence of stress CMR imaging on the management of coronary disease and its impact on prognosis.We retrospectively studied 100 consecutive patients with established CAD who underwent adenosine CMR perfusion imaging performed after a median duration of 4 months (IQR: 2.3 - 7months) after their most recent coronary procedure. We examined their hospital electronic records to determine how stress CMR imaging influenced clinical management and outcome.Mean age was 62 \u00b1 12 years, and 24% were female. 19 patients had a history of previous coronary artery bypass grafting. A stress perfusion defect in viable myocardium was identified in 71 patients (71%). Of these, 52 (73%) subsequently underwent revascularization by either PCI (n = 49) or CABG (n = 3). Revascularisation was either unsuccessful or not performed in 19 patients (27%) due to the absence of significant symptoms, presence of mild perfusion abnormality or poor revascularisation targets. Among the 29 patients in whom no inducible perfusion defects were identified, revascularization was performed in 3 patients (1%) either due to ongoing symptoms (n = 2) or presence of significant luminal area reduction on intravascular ultrasound (IVUS) (n = 1). Follow-up data was available in 93 patients (93%). During a mean follow-up of 10.2 months, no adverse events were observed in patients without inducible perfusion defects on stress perfusion CMR. Among those patients with inducible perfusion defects on CMR who were treated initially with PCI, one patient died during follow-up and four patients were referred for surgical revascularisation due to significant disease progression/restenosis post PCI.Adenosine stress CMR has significantly influenced contemporary management of coronary disease in our centre and forms an invaluable tool to assess ischaemic burden. These findings need to be explored in a larger cohort of patients with longer term follow up."} +{"text": "Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by a progressive loss of motor neurons. ALS patients, as well as animal models such as mice overexpressing mutant SOD1s, are characterized by increased energy expenditure. In mice, this hypermetabolism leads to energy deficit and precipitates motor neuron degeneration. Recent studies have shown that mutations in the gene encoding the dynein heavy chain protein are able to extend lifespan of mutant SOD1 mice. It remains unknown whether the protection offered by these dynein mutations relies on a compensation of energy metabolism defects.Cramping allele (Cra/+ mice). Dynein mutation increased adipose stores in compound transgenic mice through increasing carbohydrate oxidation and sparing lipids. Metabolic changes that occurred in double transgenic mice were accompanied by the normalization of the expression of key mRNAs in the white adipose tissue and liver. Furthermore, Dynein Cra mutation rescued decreased post-prandial plasma triglycerides and decreased non esterified fatty acids upon fasting. In SOD1(G93A) mice, the dynein Cra mutation led to increased expression of IGF-1 in the liver, increased systemic IGF-1 and, most importantly, to increased spinal IGF-1 levels that are potentially neuroprotective.SOD1(G93A) mice were crossbred with mice harboring the dynein mutant Cramping mutation in the dynein gene is, at least partially, mediated by a reversal in energy deficit and increased IGF-1 availability to motor neurons.These findings suggest that the protection against SOD1(G93A) offered by the Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by a progressive loss of motor neurons in the motor cortex, brainstem and spinal cord. ALS patients develop progressive muscle weakness and paralysis leading to death 3 to 5 years after first symptoms. Despite most cases of ALS occur sporadically, 5% are genetically inherited. Out of these familial forms of ALS, a subset is caused by mutations in the gene encoding the Cu/Zn-superoxide dismutase 1 (SOD1).Studies on patients and transgenic mutant SOD1 mice showed that ALS-linked neurodegeneration was usually associated with defects in energy homeostasis ,3. In splegs at odd angles\" (Loa), \"Cramping\" (Cra) and \"Sprawling\" (Swl) have been identified in ENU-induced mouse strains [Loa mutation disrupts, at least partially, the dynein complex [Cra/+ and Loa/+ mice are hyperactive [Cra/+ mice with striatal atrophy and compromised neurite outgrowth of striatal neurons [Cra/+ and Loa/+ mice display a major phenotype in adipose tissues [Cra/+ and Loa/+ mice show strikingly increased adipose stores, along with compromised thermogenesis. This is most likely due to defective stimulated lipolysis [Recent studies have shown that mutations in cytoplasmic dynein heavy chain gene protect motor neurons against death and extend survival of mutant SOD1 mice -8. Cytop strains ,10. All complex . In the complex . Cra/+ aeractive ,14, and neurons . Besides tissues . Indeed,ipolysis .The extension in lifespan offered by mutations in dynein heavy chain gene has been attributed to several mechanisms, including compensation of axonal transport defects and mitoCra/+ mice and studied the energetic physiology of compound heterozygotes before any obvious motor symptoms. At 16 weeks of age, SOD1(G93A) nor Cra/SOD1(G93A) mice did not show obvious clinical signs such as gait impairment but both groups of SOD1(G93A) mice were lighter than wild type littermates. Body weight deficit of Cra/SOD1(G93A) mice was less than SOD1(G93A) mice (not shown) as previously shown [Cra/SOD1(G93A) fat pads were larger than that of SOD1(G93A) mice energy deficit in early symptomatic animals.We hypothesized that the protection offered by dynein mutation against SOD1(G93A) neurodegeneration was linked to a compensation of the energy deficit of SOD1(G93A) mice. To test this hypothesis, we crossbred SOD1(G93A) mice with ly shown and bothe Figure . This waCramping dynein mutation compensated for energy deficit through decreased SOD1(G93A) linked hypermetabolism. For this, we measured energy expenditure of the four groups of mice. As previously shown [Cramping dynein mutation. Furthermore, SOD1(G93A) associated increased total energy expenditure was also unchanged and Cra/SOD1(G93A) but also Cra/+ mice were increased during the nocturnal period as compared with +/+ littermates mice was higher than that of +/+ and SOD1(G93A) mice at the same period mice mice mice mice. Indeed, spinal IGF-1 was increased in Cramping dynein mutant mice and this was further enhanced by the SOD1(G93A) transgene mice, and to increase IGF-1 levels in the spinal cord of SOD1(G93A) animals. These results, along with previous studies, suggest that two complementary protective pathways are acting in dynein mutant mice to provide the paradoxical protection towards SOD1(G93A) pathology.Our current studies provide evidence that the Loa or Cra mutation [Cra /SOD1(G93A) mice. Such a metabolic picture is fully consistent with the observed improved energy status. Thus, a straightforward interpretation would be that the dynein mutant mediated injury compensates mutant SOD1 injury through its effect on energy homeostasis, thereby alleviating neurodegeneration.This study stems from the seminal observation of a moderate lifespan extension of SOD1G93A) mice when bearing either mutation -8. These3A mice wmutation ,12. In tmutation . Furthermutation . The permutation , SOD1(G9Cra/SOD1(G93A) mice.We further provide mechanistic insights into how the dynein mutant peripheral phenotype might provide protection to motor neurons. Liver IGF-1 expression is under the control of nutritional cues, and is for instance decreased in starved animals . ConsistOur study does not directly address whether increased IGF-1 is responsible for the extension in lifespan. This is however plausible as most studies observed protective effect of increasing IGF-1 in SOD1(G93A) mice ,24. A diCra and Loa mutations were able to increase the lifespan of SOD1G93A mice. Several mechanisms have been proposed to explain the underlying mechanisms of mutant dynein protection towards mutant SOD1 pathogenesis. To conclude, we would like here to discuss critically these different hypotheses in the light of our results and provide a working model summarizing these different potential mechanisms.Several research groups, including ours, have previously shown that dynein In the first study, Kieran et al. suggesteLegs at odd angles and Cramping, also impact bioenergetics' mechanisms and revert the energy deficit of SOD1(G93A) mice. In the compound mice, this leads to increased production of IGF-1. We propose that the proprioceptive degeneration and CNS linked hyperactivity converge to modify neuronal activity and facilitate the net transcytosis of IGF-1 in the vicinity of motor neurons, thus achieving neuroprotection. This working model is summarized in Figure The complex phenotype of dynein mutant mice render very plausible that dynein mutation could also act at multiple levels besides motor neurons. First, dynein mutation is likely to reduce glutamate excitotoxicity through the degeneration of proprioceptive glutamate inputs to motor neurons . AlthougCramping mutation in the dynein gene is, at least partially, mediated by a reversal in energy deficit and increased IGF-1 availability to motor neurons. Our study provides a rationale explanation to increased survival in double dynein mutant/SOD1(G93A) mice and suggests that the neuroprotection results from dynein mutant phenotype in various tissues.These findings suggest that the protection against SOD1(G93A) offered by the Cra/+ females were crossed with SOD93A males and were identified by tail DNA genotyping for the human transgene SOD1 (G93A) and the Cra mutation as described previously [ad libitum. All animal experiments were performed under the supervision of authorized investigators and followed current EU regulations. Animals were treated in accordance with the European Union guide for the care and the use of animals in research . LD has been allowed to perform mouse experiments .Heterozygous eviously ,9. Exper2 consumption and CO2 production by using an open-circuit indirect calorimetry system . Concentrations of O2 and CO2 in the outgoing air were successively measured in five different cages. The system was rinsed for 90 s between each measurement. Final values of gas concentrations were the mean of 10 measures obtained during 40 s. Each cage was sampled every 11 min, and one cage was left vacant as reference of ambient gas concentrations. Measurements were performed continuously over 23 1/2 h, a 30-min period being required for calibration of the O2 and CO2 analyzers. In total, 127 measures were collected per day and mouse. The average of the five lowest values of O2 consumption was considered as resting energy expenditure. Energy expenditure was obtained by using an energy equivalent of 20.1 J/ml O2. The respiratory quotient was the ratio of CO2 production over O2 consumption.We measured O\u00ae . Real time RT quantitative PCR was performed with one microgramme of total RNA as described [RNA were extracted by using Trizolescribed . PCR anaMouse IGF-1 levels were measured using Mouse/Rat IGF-I Quantikine ELISA Kit (R&D systems), using manufacturer's instructions. Plasma triglycerides and NEFAs were measured using Randox kits using manufacturer's instructions.Statistical comparisons were accomplished with the unpaired Student t test, unless otherwise indicated, or ANOVA followed by the post hoc Newman-Keuls multiple comparisons test using PRISM version 2.0a software .Cra: Cramping mutation of the Dyn1hc1 gene; FAS: fatty acid synthase; IGF-1: Insulin-like growth factor 1; LPL: lipoprotein lipase; SOD1: copper-zinc superoxide dismutase.AchR\u03b1: alpha subunit of the nicotinic cholinergic receptor; ALS: amyotrophic lateral sclerosis; BBB: blood brain barrier; BicD2: bicaudal D2; CNS: central nervous system; The authors declare that they have no competing interests.AF performed animal handling and follow up, participated in the indirect calorimetry experiments, performed qPCR assays and most of the biochemical assays; JE performed qPCR assays and some biochemical assays, analyzed indirect calorimetry results and drafted the manuscript; HO performed indirect calorimetry experiments; YL performed animal follow up and analyzed results; BS performed animal handing, genotyping and analyzed results; ACL and JPL conceived of the study, participated in its design and helped to draft the manuscript; LD conceived of the study, designed and coordinated the study and drafted the manuscript. All authors read and approved the final manuscript."} +{"text": "Incidence of urinary tract infections is elevated in patients with diabetes mellitus. Those patients show increased levels of the saturated free fatty acid palmitate. As recently shown metabolic alterations induced by palmitate include production and secretion of the pro-inflammatory cytokine interleukine-6 (IL-6) in cultured human bladder smooth muscle cells (hBSMC). Here we studied the influence of palmitate on vital cell properties, for example, regulation of cell proliferation, mitochondrial enzyme activity and antioxidant capacity in hBSMC, and analyzed the involvement of major cytokine signaling pathways.HBSMC cultures were set up from bladder tissue of patients undergoing cystectomy and stimulated with palmitate. We analyzed cell proliferation, mitochondrial enzyme activity, and antioxidant capacity by ELISA and confocal immunofluorescence. In signal transduction inhibition experiments we evaluated the involvement of NF-\u03baB, JAK/STAT, MEK1, PI3K, and JNK in major cytokine signaling pathway regulation. We found: (i) palmitate decreased cell proliferation, increased mitochondrial enzyme activity and antioxidant capacity; (ii) direct inhibition of cytokine receptor by AG490 even more strongly suppressed cell proliferation in palmitate-stimulated cells, while counteracting palmitate-induced increase of antioxidant capacity; (iii) in contrast knockdown of the STAT3 inhibitor SOCS3 increased cell proliferation and antioxidant capacity; (iv) further downstream JAK/STAT3 signaling cascade the inhibition of PI3K or JNK enhanced palmitate induced suppression of cell proliferation; (v) increase of mitochondrial enzyme activity by palmitate was enhanced by inhibition of PI3K but counteracted by inhibition of MEK1.Saturated free fatty acids cause massive alterations in vital cell functions of cultured hBSMC involving distinct major cytokine signaling pathways. Thereby, certain cytokines might counteract the palmitate-induced downregulation of cell proliferation and vitality. This could be an important link to clinical findings of increased risk of metabolic related bladder diseases such as overactive bladder (OAB) and bladder pain syndrome/interstitial cystitis (BPS/IC). Abnormal urinary storage symptoms are linked to obesity and poor general health Palmitate is an inflammatory stimulus and can induce IL-6 and MCP-1 expression and secretion in cultured human bladder smooth muscle cells Various studies have shown palmitate dependent regulation of cytokines such as IL-6 Since basic bladder function depends on balanced cellular interactions, cytokines are a major regulation factor in normal and pathological bladder states Therefore we investigated the effect of palmitate onto four major pathways involved in cytokine signaling: (1) janus kinase (JAK) activation of Signal transducers and activators of transcription (JAK/STAT); (2) Phosphoinositol-3-kinases (PI3K); (3) Mitogen-activated protein kinase kinase 1 (MEK1); (4) c-Jun N-terminal kinase (JNK).The JAK/STAT pathway regulates vital cell processes such as cell proliferation, survival and inflammation PI3K pathway is involved in cytokine modulation of cell proliferation, cell motility and cell survival MEK1 regulates activation of extracellular signal-regulated kinase (ERK) JNK is a mitogen-activated protein kinase, which regulates gene transcription via cJUN activation, associated with oxidative stress and inflammation In the present study we investigated palmitate effects on vital cellular functions of cultured human bladder smooth muscle cells and their modulation by inhibition of some major cytokine signaling pathways. We found that palmitate inhibited cell proliferation, and increased mitochondrial activity and antioxidant capacity. Those functions are differentially modulated by inhibition of major cytokine signaling pathways.The study was approved by the Ethics Committee of the University of Leipzig (Reg. No. 773) and was conducted according to the principles expressed in the Declaration of Helsinki. Written informed consent was obtained from all patients.Human bladder smooth muscle cell (hBSMC) cultures were established from human detrusor muscle after radical cystectomy of tumor patients as described earlier For pathway analyses, cells were pre-incubated for 1 h with 40 \u00b5M MG132 , 2 \u00b5M AG490 , 20 \u00b5M PD98059 in serum free medium. For transfection HiPerFect Transfection Reagent (Qiagen) were used. As negative, non-silencing control the AllStars Negative Control siRNA, including in this kit, was used. The Negative Control siRNA was labeled with Alexa Fluor 488 fluorescence dye, which allowed monitoring of transfection efficiency by LSM-5 Pascal confocal laser scanning microscope . SOCS3 knockdown efficiency was detected by quantitative PCR after 24 h of transfection.Total RNA was isolated with AllPrep DNA/RNA/Protein Mini Kit (Qiagen). Quantitative PCR was performed with the real-time PCR-System realplex2 Mastercycler using the SYBR-Green quantitative PCR Mastermix and custom primers . 2 \u00b5g total protein were transferred in triplicates on nitrocellulose membrane by Dot Blotting . After blocking with Odyssey blocking buffer for 1 h the membranes were incubated with anti-SOCS3 rabbit IgG over night at 4\u00b0C. For detection we used anti-rabbit IRDye 680 for 2 h. Membranes were scanned with Odyssey Infrared Imager and evaluated by Odyssey Infrared Imaging Software 3.0 (Licor Biosciences). Total protein was visualized by SYPRO Ruby blot stain .For cell proliferation and mitochondrial enzyme activity analysis cells were used at 80% confluency in 96-well plates and incubated over night.Cell proliferation was measured by BrdU colorimetric cell proliferation ELISA by quantifying BrdU incorporated into the newly synthesized DNA of replicating cells.Mitochondrial enzyme activity was analyzed using 3--2,5-diphenyltetrazolium bromide (MTT) assay. MTT is reduced to purple formazan in living cells. Hydrochloric acid is added to dissolve the insoluble purple formazan product into a colored solution. The absorbance of this colored solution can be quantified by measuring at 570 nm by a spectrophotometer.The antioxidant capacity was measured by Antioxidant Assay , which show the cumulative effect of all antioxidants present in cell lysate include enzymes , macromolecules and small molecules .Secreted IL-6 was quantified by Quantikine human IL-6 immunoassay . IL-6 in cell culture supernatants were measured after being centrifuged in 1\u2236100 diluted samples.Cells were cultured on cover slips. At 80% confluence cells were fixed in 4% buffered paraformaldehyde. For detection of protein carbonylation cells were pre-incubated 1 h with 0.1% 2,4-dinitrophenyl hydrazine which reacts with protein carbonyls. Cells were incubated over night at 4\u00b0C with primary antibodies for DiniComplete data analysis was performed using Prism 5.0 statistical software. The data are presented as the mean +/\u2212 SEM from at least six independent experiments. Statistical differences were analyzed by ANOVA. A P-value <0.05 was considered statistically significant.Treatment of hBSMC with FFA palmitate (up to 48 hrs) resulted in significant decrease of cell proliferation in a time dependent manner . FurtherIt is well known that palmitate induces NF-\u03baB via protein kinase-C (PKC)-activity and that NF-\u03baB release can be blocked by MG132, a potent inhibitor of proteasomal degradation of IkB In gene expression experiments we found down-regulation by NF-\u03baB of genes related to cell proliferation we investigated the JAK/STAT receptor signaling using AG490 for direct inhibition of the cytokine receptor, and we used SOCS3 inhibition to release full STAT3 signaling cascade, (ii) we analyzed the signalling downstream JAK/STAT. Palmitate stimulation under direct inhibition of the cytokine receptor by the specific JAK/STAT inhibitor AG490 significantly decreased cell proliferation and antioxidant capacity while mitochondrial activity was unaltered . This waAs illustrated in Downstream of pSTAT3, the IL-6-JAK/STAT pathway can be regulated by MEK 1, PI3K and JNK signaling . InhibitA more prominent regulation of cellular response was observed when phosphoinositol (PI)-3-kinase signaling was inhibited (LY294002). Palmitate reduced cell proliferation was dramatically further inhibited when palmitate was applied together the PI3K inhibitor LY294002 . MitochoOn gene expression level inhibition of PI3K during palmitate stimulation showed varying effects on the examined genes . We founAnother major downstream pathway of cytokine signaling is characterized by its key-enzymes c-Jun N-terminal kinases JNKs; . SelectiIn the present study we were interested in the effect of palmitate on various cellular functions, on gene expression related to vital cell properties, and the involvement of the major cytokine signaling pathways into palmitate stimulated effects. We investigated the modulation of the cytokine receptor signaling cascade always related to palmitate stimulation and therefore we did not evaluate the effects of inhibitors alone on naive cells, which may be some limitation of the study regarding the analysis of cytokine receptor induced signaling cascades proper.Several studies have consistently demonstrated that elevated levels of plasma FFA, such as palmitate, were found in obesity related disorders Palmitate inhibits cell proliferation in hBSMC in time dependent manner and simuBesides TLR4 mediated NF-\u03baB release , NF-\u03baB cInhibition of NF-\u03baB by the proteasome inhibitor MG132 enhanced these effects , Fig. 2AReduced cell proliferation might influence the capability of tissue remodeling and regeneration following insults to the urinary bladder. However, up to date data on morphological changes in detrusor smooth muscle cells in overactive bladder and other bladder dysfunctions are sparse. Most studies indicate alterations of basic smooth muscle cell functions on a cellular basis, i.e. cell-cell communication In addition, our studies show that elevated palmitate levels lead to increased mitochondrial enzyme activity and antioxidant capacity in hBSMC . Indeed,We here provide evidence that palmitate modulates protein carbonylation and lipid peroxidation . PalmitaLipid peroxidation, which is known to impair lipid function, is another major direct effect of ROS and enhanced antioxidant capacity should prevent lipid peroxidation. However, we found increased staining for malone dialdehyde MDA, followinFor the first time our study reveals important insights, that elevated levels of FFAs cause the increased expression of peroxisome proliferator\u2013activated receptor (PPAR \u2013coactivator-1 (PGC-1) , a transMultiple intracellular pathways could be involved in the regulation of proliferation, apoptosis induction, mitochondrial activity and increase of antioxidant capacity. When blocking NF-\u03baB signaling by inhibition of proteasome degradation of phosphorylated IkB-P during palmitate stimulation cell proliferation and antioxidant capacity was suppressed, while mitochondrial activity was not affected , Fig. 2.In our previous studies, the free fatty acid palmitate induced time and concentration dependent IL-6 release in hBSMC, followed by autocrine and paracrine regulation of the IL6 pathway Studies investigating cytokine signaling in human bladder smooth muscle cells are rare; most studies have investigated cytokine induced IL-6 release by hBSMC Previously, we could show IL-6 release after palmitate stimulation . The IL-Direct cytokine signaling can be analyzed either by inhibition of the JAK/STAT3 inhibitor SOCS3 To identify the regulation of cell proliferation downstream JAK/STAT we inhibited MEK1 by PD98059, JNK by SP600125 and PI3K by LY294002 . Both PIIn our experiments the eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) was down-regulated by inhibition of PI3K . PI3K/AKFurthermore, inhibition of JNK leads to increased BCL-XL gene transcription a gene, Since those pathways are especially activated by IL-6, it seems that this cytokine may have an overall beneficial effect in case of cell deterioration. Other cytokines may, however, show divergent effects, depending on the degree of their activation of certain downstream pathways.Our study indicates that high levels of the saturated free fatty acid palmitate induce down regulation of cell proliferation through NF-\u03baB and the formation of reactive intermediates leading to programmed cell death. Fatty acid-induced mitochondrial and antioxidant activity may contribute to human bladder disorders related to obesity. Additionally, palmitate-mediated production of ROS may cause significant cellular dysfunction that could contribute to the pathogenesis of those diseases without massive tissue reorganization due to programmed cell death. Cytokine signaling is involved in the process of cell proliferation and regulation of genes related to mitochondrial and antioxidant activity. Our findings suggest that metabolic and endocrine status is an important factor for the pathogenesis of bladder dysfunction. Pharmacologic approaches targeting and aiming to restore cytokine network in the bladder could be a novel therapeutic option.Figure S1Palmitate effect on cell proliferation. Phase contrast images of cultured hBSMC after 24 h and 48 h stimulation with 0.25 mM palmitate. Inset represents 2 fold magnification showing cellular alterations by 48 h stimulation with 0.25 mM palmitate: membrane blabbing (arrows) and detaching of cells (arrowheads).(TIF)Click here for additional data file.Figure S2Alteration in IL-6 secretion of hBSMC. Cultured hBSMC were pre-incubated 1 h with cytokine signaling pathway inhibitors and following stimulated 48 h with 0.25 mM palmitate (black column). IL-6 secretion was measured using ELISA and data are shown as mean + SEM from three different cultures. Significant differences related to palmitate are indicated by bars.(TIF)Click here for additional data file.Figure S3Palmitate influence pSTAT3 expression. Confocal immunofluorescence of pSTAT3 (green) in cultured hBSMC after 48 h stimulation with 0.25 mM palmitate and palmitate plus NF-\u03baB inhibitor MG132. Nuclei were stained with DAPI (blue). The inset represents negative staining control without using primary antibody against pSTAT3.(TIF)Click here for additional data file."} +{"text": "Infections, new onset diabetes mellitus (NODAT) and rejection are frequent complications after renal transplantation and may be related to innate immunity alterations. We evaluate the relationship between serum mannose-binding lectin (MBL) levels, chronic inflammation, infection, NODAT and subclinical rejection after renal transplantation.Between March 2005 and October 2006 consecutive non-diabetic renal transplant recipients were recruited. Serum levels of MBL, soluble tumor necrosis factor receptor 2 (sTNFR2) and neutrophil gelatinase associated lipocalin (NGAL) were determined before transplant and at 1 and 3 months. An oral glucose tolerance test was performed at 3 months. A surveillance 3-month renal biopsy was performed in a subset of 60 patients with stable renal function.A total of 125 patients were recruited and 111 had a functioning graft at 3 months. MBL serum levels remained unchanged following transplantation. Subjects with low MBL (lower tertile) had higher pretransplant sTNFR2 and NGAL , an increased incidence of bacterial/fungal infection (p=0.021) and an increased prevalence of NODAT at 3 months . Multivariate analysis confirmed that MBL was a risk factor for NODAT adjusting for age, pre-transplant impaired fasting glucose and body mass index. Subclinical rejection in the 3-month surveillance biopsy was observed in 7 of 18 (38.9%) low MBL patients and in 3 of 42 (7.1%) high MBL patients (p=0.005). Induction and maintenance immunosuppression was not different in patients with low and high MBL levels.Low MBL serum levels in renal transplants are associated with major outcome variable after renal transplantation such as bacterial/fungal infections, NODAT and subclinical rejection. These results suggest that alterations of the innate immunity may play an important role in renal transplantation."} +{"text": "The emergence of pandemic (H1N1) 2009 virus highlighted the need for enhanced surveillance of swine influenza viruses. We used real-time reverse\u2013transcription PCR\u2013based genotyping and found that this rapid and simple genotyping method may identify reassortants derived from viruses of Eurasian avian-like, triple reassortant-like, and pandemic (H1N1) 2009 virus lineages. Co-infection of influenza A viruses enables viral gene reassortments, thereby generating progeny viruses with novel genotypes. Such reassortants may pose a serious public health threat, as exemplified by the emergence of pandemic influenza (H1N1) in 2009 2009 virus or virus segments that are closely related to this lineage from triple reassortant swine linage . We also tested various probe and SYBR green concentrations under different PCR conditions. The condition described above gave the most robust and consistent DNA amplification (data not shown). We tested 31 human pandemic (H1N1) 2009 and 63 human seasonal influenza viruses as controls. As expected, all human pandemic influenza viruses were double positive and all seasonal influenza samples were double negative in all 8 assays.2 to 108 copies/reaction (2 to 108 copies/reaction (data not shown). However, to avoid nonspecific SYBR green signals, we purposely limited the number of amplification cycles to 30.To evaluate the sensitivity of the assays, we tested serial diluted plasmid DNA of the corresponding segments of influenza A/California/4/2009 virus as a standard. The fluorescent signals generated from the SYBR green reporter dye in all assays were highly similar to those previously reported (reaction . As expereaction (9). ThiUsing these assays, we tested 41 swine virus isolates collected during January 2009\u2013January 2010. In all 8 reactions, 10 pandemic (H1N1) 2009 virus samples transmitted from humans to pigs . In thesTo demonstrate the potential use of these assays in studying swine viruses circulating in other geographic locations, we tested 7 recent swine isolates collected in the United States. Genotyping results agreed 100% with data deduced from sequence analyses . We alsoThe emergence of pandemic (H1N1) 2009 has highlighted the need for global systematic influenza surveillance in swine. Our results demonstrated that the addition of locked nucleic acid hydrolysis probes specific for pandemic (H1N1) 2009 virus into previously established SYBR green assays can help differentiate segments of pandemic (H1N1) 2009, Eurasian avian-like, and triple reassortant virus lineages. These assays might provide a rapid and simple genotyping method for identifying viruses that need to be fully genetically sequenced and characterized. They may also help provide better understanding of the viral reassortment events and viral dynamics in pigs. Although at present, genes derived from human seasonal viruses cannot be characterized with our modified assays, the performance of our assays warrants similar investigations for genotyping human influenza viruses.Genotyping results , viral sequences, and Combined SYBR green/hydrolysis probe quantitative RT-PCR assays for swine influenza virus"} +{"text": "RP2) gene are associated with X-linked RP, which is a phenotypically heterogenic form of retinal degeneration. The purpose of this study was to assess the functional consequence of disease-associated mutations in the RP2 gene using an in vivo assay. Morpholino-mediated depletion of rp2 in zebrafish resulted in perturbations in photoreceptor development and microphthalmia . Ultrastructural and immunofluorescence analyses revealed defective photoreceptor outer segment development and lack of expression of photoreceptor-specific proteins. The retinopathy phenotype could be rescued by expressing the wild-type human RP2 protein. Notably, the tested RP2 mutants exhibited variable degrees of rescue of rod versus cone photoreceptor development as well as microphthalmia. Our results suggest that RP2 plays a key role in photoreceptor development and maintenance in zebrafish and that the clinical heterogeneity associated with RP2 mutations may, in part, result from its potentially distinct functional relevance in rod versus cone photoreceptors.Genetic mutations are frequently associated with diverse phenotypic consequences, which limits the interpretation of the consequence of a variation in patients. Mutations in the retinitis pigmentosa 2 ( Recent advances in whole genome sequencing and exome capture techniques have exploded the field of identification of new disease in patients with genetic diseases Degeneration or dysfunction of photoreceptors is frequently associated with variable clinical presentation, likely due to their unique structure and metabolic demands Retinitis Pigmentosa (RP) represents one such disorder that exhibits both clinical and genetic heterogeneity in patients RP2 gene (NM_006915.2)encodes a polypeptide of 350 amino acids The We and others have reported the association of wide-spectrum of clinical phenotype in patients with RP2 mutations. These clinical features include severe to late-onset disease, typical RP, as well as macular dystrophy It has been shown that analysis of zebrafish embryos provides an excellent platform to analyze in vivo effect of silencing of genes followed by disease progression and pathogenesis rp2 function, we designed a translation blocking (AUG) morpholino against the zebrafish rp2 gene (NM_213446.1) and injected into 2\u20134 cells stage wild-type (WT) zebrafish embryos rp2-MO injected embryos (\u223c15% of defective embryos) and abnormal photoreceptor development in a majority (\u223c90%) of embryos, although retinal development seemed to proceed normally, at least up to 5 dpf. However, injection of higher doses of the rp2-MO resulted in defective retinal lamination (data not shown). Measurement of the eye diameters on the histological sections indicated approximately 32% reduction in size of defective embryo eyes compared to control embryos compared .rp2 results in early developmental defects in zebrafish embryos (data not shown).Histological analysis of zebrafish retinas at 4 days post fertilization (dpf) further revealed very few to no outer segment development in the defective embryos . Disruptrp2 on the expression and localization of photoreceptor-specific proteins in the embryo retinas. To this end, we performed immunofluorescence analysis of control and rp2-MO treated 4 dpf embryo retina using Zpr1 and Zpr3 antibodies. As shown in rp2 results in a loss of both rod and cone photoreceptor specific staining pattern, which also indicates a defect in photoreceptor morphology at its carboxyl-terminus. We found that the human mRNA could efficiently rescue the retinopathy phenotype associated with suppression of rp2, whereas mRNA encoding GFP alone did not result in a rescue. Histological and immunofluorescence analyses revealed preservation of photoreceptor morphology and expression of rod and cone-specific proteins in the retina of embryos treated with mRNA encoding RP2-GFP fusion protein. As shown in rp2-MO treated embryos injected with mRNA encoding GFP alone did not show OS development and expression of Zpr1 or Zpr3 but not in cone photoreceptors .www.Zfin.org. Rhodopsin 1D4 antibody was a kind gift of Dr. Robert S. Molday.Rabbit anti-RP2 antibody was generated by immunizing rabbits with full length recombinant human GST-RP2 and characterized for specificity, as described Histological analysis on zebrafish retina was performed on JB4 plastic sections as previously described [38]. Paraformaldehyde fixed embryo were analyzed for immunofluorescence as described earlier. Electron Microscopy was done using standard protocols. Briefly, zebrafish were fixed in perfused with 4% paraformaldehyde and 2.5% glutaraldehyde in PBS. The fish were treated with osmium tetroxide and dehydrated using graded ethanol series followed by treatment with propylene oxide and embedded in Epon-812 resin. Gold interference (50\u201380nM) sections mounted on grids were post stained uranyl acetate (5% in ethanol), and lead citrate (4% aqueous solution), washed with water and dried before imaging under Philips C10 TEM at 80 KV.RP2 gene was cloned into pEGFP-N1 (Clonetech) vector and mutations were created using QuickChange mutagenesis kit (Stratagene). Capped mRNA was synthesized in vitro with mMessenger mMachine T7 kit (Ambion).To knock down zebrafish RP2, we synthesized a translation blocking morpholino (Gene-Tools) (AUG-MO 5\u20323\u201d) (Danio rerio) were reared and maintained as previously described Zebrafish Figure S1RP2 is expressed in both rod and cone photoreceptors. Immunofluorescence analysis of 4 dpf zebrafish embryos injected SNC-MO and rp2-MO was performed using anti-rhodopsin (1D4) antibody or PNA (red). Nuclei are stained with DAPI (blue). RPE: retinal pigmented epithelium; OS: outer segment; IS: inner segment; ONL: outer nuclear layer; INL: inner nuclear layer; GCL: ganglion cell layer.(TIF)Click here for additional data file."} +{"text": "TSH-secreting tumours are extremely rare case of hyperthyroidism. Most important clinical feature is preserved TSH level in subjects with apparent thyrotoxicosis. Possible misdiagnosis of primary thyroid hyperfunction could lead to mistreatment with anti-thyroid medications. This worsens disease course and outcome. Neurosurgery success rate is limited by tumour size and its extrasellar expansion. Native somatostatin is key negative regulator of TSH secretion. In most cases tumour cells express somatostatin receptors. This feature creates potential use of somatostatin analogues for medical treatment of TSH-secreting tumours.The aim was to determine potential value of somatostatin analogues in primary TSH-oma treatment. Secondary objective was to evaluate efficacy of long-acting somatostatin analogues in cases after unsuccessful neurosurgery.Material comprised of 17 patients with secondary thyrotoxicosis, 7 women and 10 men, aged 20 to 69 years (mean 39), presenting with pituitary macroadenoma (16) and one with microadenoma and empty sella. Before diagnosis was established 8 out of 17 patients received antithyroid medication, in 2 cases strumectomy was performed and 2 patients received 131I therapy.Somatostatin analogue octreotide long acting repeatable (LAR) administration at least 3 months (3 injections) before surgery, and in cases of unsuccessful surgery stable octreotide LAR therapy.Initially, all patients had elevated fT4 and \u03b1-SU levels . 3 months of octreotide LAR therapy led to significant fT4 reduction (to mean 12.2 pmol/l) and TSH reduction from mean 6.5 mIU/ml to 0.3 mIU/l). In all cases clinical improvement was observed.In 14 out of 17 pre-treated with octreotide LAR decreased tumour volume and in 2 improvement in visual field was observed.Patients in euthyroid state were referred to neurosurgery department. Transsphenoidal adenomectomy was successful 13 out of 17. In four cases after unsuccessful neurosurgery intervention stable euthyroidism was achieved with octreotide LAR treatment. In one case secondary thyrotoxicosis relapsed 2 years and now is controlled by octreotide treatment.Somatostatin analogue treatment is efficient in TSH-secreting tumours in the terms of TSH secretion, thyroid function and clinical improvement. In cases of surgery failure prolonged octreotide treatment could be safe and efficient option of disease management."} +{"text": "Lynch syndrome is characterized by germline mutations in DNA mismatch repair (MMR) genes and carries up to a 70% lifetime risk of colorectal cancer. Impaired MMR gene function results in an abundance of small aberrant nucleotide repeat sequences termed microsatellite instability (MSI). MSI is present in 80-85% of colorectal cancers associated with Lynch syndrome. Prior studies have demonstrated that MMR gene function (as measured by immunohistochemistry) is often lost in Lynch-associated adenomas but is preserved in hyperplastic polyps. The aim of our study was to evaluate the prevalence of MSI in adenomatous and hyperplastic polyps from individuals with Lynch Syndrome.We identified 63 polyps (37 adenomas and 26 hyperplastic polyps) from 34 subjects with known germline MMR gene mutations. Clinicopathological information for each polyp was obtained from retrospective review of pathology reports. MSI analysis was performed on microdissected polyp DNA using pentaplex PCR for a panel of five quasimonomorphic mononucleotide repeat sequences . If \u22652/5MSI was identified in 14/37 (38%) adenomas, compared with 3/26 (12%) hyperplastic polyps (p = 0.021). Prevalence of MSI was significantly higher in larger polyps; among the lesions \u226510mm, 6/7 (86%) adenomas and 1/1 (100%) hyperplastic polyps demonstrated MSI. There was no association between MSI status and patient sex, polyp location, or the patient\u2019s underlying MMR gene mutation. Although MSI appeared more prevalent in adenomas from individuals \u2265age 50 years (53% vs 22% for age<50), this did not achieve statistical significance (p = 0.057).Overall, MSI was detected in 38% of adenomas and 12% of hyperplastic polyps from individuals with MMR mutations. MSI analysis of small colorectal adenomas would have low sensitivity for identifying patients who should be considered for genetic testing for Lynch syndrome. The finding of MSI in a small fraction of hyperplastic polyps raises questions about their neoplastic potential in patients with Lynch syndrome."} +{"text": "PEEP selection during mechanical ventilation (MV) for patients with acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) remains a challenge for clinicians. Clinicians often rely on experience and intuition in setting MV, resulting in a more variable treatment and outcome. We hypothesise that monitoring patient-specific respiratory system elastance (Ers) during PEEP change may provide an insight into the patient's condition.2O) recruitment manoeuvre (RM) until peak airway pressure reaches 45 cmH2O. Airway pressure and flow profile were recorded using a pneumotachometer. The change of patient's respiratory system elastance (Ers = 1/compliance) and the end of expiratory lung volume (EELV) during RM were estimated and studied. The trials were approved by New Zealand South Island Regional Ethics Committee.Thirteen patients with ALI/ARDS underwent a step-wise PEEP increase (5 cmH2O/l (26.1 to 51.0), reflecting the heterogeneity of the patients and their response to PEEP. This outcome supports the idea that MV/PEEP should be individualised. During RM, patients' Ers decreased with PEEP increase until a specific minimum and increase at higher PEEP. The decreased of Ers suggest alveolar recruitment whereas an increase of Ers at higher PEEP shows potential overinflation. An example is shown in Figure The median (IQR) Ers over all patients was 34.0 cmHThe change of patient-specific Ers and EELV during minimally invasive PEEP titration provides an insight into the patient's lung condition, and thus could potentially be used as a method to individualise MV treatment and, in particular, PEEP selection."} +{"text": "Idiopathic intracranial hypertension (IIH) typically manifests with headaches and visual disturbance. Unusual presenting features of IIH are sometimes described. Hemi facial spasm (HFS) in women has an incidence of 0.81 per 100,000 compared to an IIH incidence rate of 3.5-20 per 100,000. We describe the 4th case of HFS induced by IIH and we propose a new nomenclature.We present a case of a 33 year old right handed woman with sequential new onset headache causing secondary HFS. Neurological examination revealed bilateral papillaoedema and confirmed right sided facial spasms. Cerebrospinal fluid examination showed an opening pressure of 33cm H2O and normal CSF constituents. High resolution brain MRI and MRV were normal including no evidence of neurovascular compression in the cerebello-pontine angle. EEG was entirely normal. The headache and HFS resolved after lowering CSF pressure. At three months post lumbar puncture she remained asymptomatic with no further episodes of HFS. Conclusion: There are only three prior cases of IIH associated with HFS in the published literature. We describe a further case of a very rare clinical manifestation of IIH and propose a possible pathophysiological mechanism and a new name for this entity \u201cIIH-Spasm syndrome\u201d."} +{"text": "The aim of the study was to assess (1) the feasibility and the outcomes of secondary neck dissections in well differentiated, medullary and poorly differentiated thyroid cancer and (2) to evaluate complications associated with surgical treatment.We assessed the results of secondary neck dissections in 51 patients previously operated for thyroid cancer: 33 well differentiated thyroid cancer, 15 medullary cancer, 3 poorly differentiated thyroid cancer.Reoperations covered I-VII neck levels. Radical neck dissection was performed in 20 patients, selective neck dissection (SND) in 31 patients. These included 16 central compartment (CC) and 10 mediastinal excisions. Postoperative complications were stated in 13 patients: 4 chyle leaks, 3 massive bleedings, 8 injuries of RLN, hypoparathyroidism in 22 patients, 2 patients died in perioperative period. Stimulated Tg<2mg/ml was observed in 7 patients with WDTC during the first control after neck dissection; in 6 patients Tg level decreased after operation; 7 patients had still notably elevated Tg levels (>30 ng/ml). None of the patients with medullary cancer achieved calcitonin level lower than 10 pg/ml; 9 patients developed distant metastases.Patients with nodal metastases deriving from thyroid cancer present a challenging group for surgeons. The policy is to operate due to strong indications. It is important to be aware of possible complications. The outcomes of neck dissections in patients with medullary and poorly differentiated thyroid cancer were unsatisfactory."} +{"text": "Shocked patients with pelvic fractures are amongst the most severely injured patients requiring trauma care. They present a complex management challenge with mortality rates of up to 60%, the majority being the result of early exsanguination. Errors aWe analysed the clinical care and PIP activities of all adult patients presenting to our Major Trauma Centre with exsanguinating pelvic trauma, for a period of 4 years (2007 \u2013 2010) following the incorporation of the American College of Surgeons Trauma PIP into locOne hundred eighty-five shocked patients with pelvic fractures were included. Median Injury Severity Score was 34 and median base deficit was 8.25 mEq/L. Sixty-two patients (34%) died from their injuries. The PIP identified pitfalls in the structure or process of care in one third of deaths. The most common were \u2018errors in decision-making leading to delays or inappropriate treatment\u2019 (32%) and \u2018delays in blood or blood product availability\u2019 (27%). Implemented changes included: 1) a decision-making algorithm for the management of exsanguinating pelvic trauma, 2) a massive haemorrhage protocol and 3) appointment of specialist pelvic orthopaedic surgeons. These changes were associated with significant annual improvements in the targeted processes of care, including: time to massive haemorrhage protocol activation (p=0.04); volume of crystalloid transfused (p<0.01); ratios of blood to blood-products transfused (p<0.0001); time to primary surgical haemorrhage control (p=0.05) and proportion of patients undergoing definitive pelvic fixation (p=0.01). Survival improved from 45% (2007) to 79% (2010) with a significant annual reduction in mortality (p=0.002) that remained after adjusting for baseline patient and injury characteristics. This trend was not evident nationally.The institution of a PIP allowed the identification and targeted improvement of aspects of trauma care that impact outcome and resulted in rapid and sustained mortality reductions in this severely injured patient group."} +{"text": "After the publication of our article , we founThe mathematical definition of expanded temporal block given in Definition 3 should have been \"the corresponding expanded temporal block And the subscript of some variable involved in partial correlation calculation (formula (2)) given in Definition 4 should have been \"X, Z\" rather than \"X, 2\". The corrected formula is shown here:We regret any inconvenience these errors may have caused.The authors declare that they have no competing interests."} +{"text": "OsGSTU4. OsGSTU4 fusion protein was found to be localized in nucleus and cytoplasm. The over-expression of OsGSTU4 in E. coli resulted in better growth and higher GST activity under various stress conditions. Further, we raised over-expression transgenic Arabidopsis plants to reveal its in planta function. These transgenic lines showed reduced sensitivity towards plant hormones, auxin and abscisic acid. Various analyses revealed improved tolerance in transgenic Arabidopsis plants towards salinity and oxidative stresses, which may be attributed to the lower accumulation of reactive oxygen species and enhanced GST activity. In addition, microarray analysis revealed up-regulation of several genes involved in stress responses and cellular detoxification processes in the transgenic plants as compared to wild-type. These results suggest that OsGSTU4 can be used as a good candidate for the generation of stress-tolerant crop plants.Glutathione S-transferases (GSTs) are multifunctional proteins encoded by large gene family in plants, which play important role in cellular detoxification of several endobiotic and xenobiotic compounds. Previously, we suggested the diverse roles of rice GST gene family members in plant development and various stress responses based on their differential expression. In this study, we report the functional characterization of a rice tau class GST gene, Abiotic stresses are the major environmental factors affecting crop production worldwide by reducing growth and productivity of plants. Exposure of plants to various abiotic stresses leads to oxidative damage Glutathione S-transferases (GSTs) are ubiquitous enzymes encoded by a large family of genes, which play an important role in cellular detoxification to a wide variety of endobiotic and xenobiotic substrates by conjugating the tripeptide glutathione 2O2GSTs have been found to be differentially regulated by various abiotic stresses, including dehydration OsGSTU4 (Genbank accession number FN428745). We raised and analyzed Arabidopsis transgenic plants over-expressing OsGSTU4. Our results showed increased tolerance in the transgenic plants to salinity and oxidative stresses at various developmental stages.In our previous study, we observed preferential expression of GST genes in various developmental stages, and differential expression in response to various stresses Arabidopsis thaliana ecotype Columbia was used as wild-type (WT) for all experiments and genetic background for the generation of transgenic plants. Seeds were obtained from Arabidopsis Biological Resource Centre, USA. Plants were grown in plates containing MS medium and in pots containing vermiculite supplemented with liquid Arabidopsis nutrient medium in a culture room under 80\u2013100 \u03bcmol m\u22122 S\u22121 at 22\u00b11\u00b0C with 14/10 h of day/night photoperiod, as described previously For gene expression analysis in rice, tissue samples from various developmental stages, hormone treatments, and abiotic stress conditions were used as described previously Escherichia coli, the complete open reading frame (ORF) of OsGSTU4 was amplified by PCR using gene-specific primers (TAAGGATCCATCCATGGCCGGTGGAG and GCAAAGCTTTCAGTTGGTGGCAG) and cloned into the expression vector, pET28a, in BamHI and HindIII restriction sites. The resulting recombinant plasmid, pET28a:OsGSTU4 was transformed into E. coli strain BL21 (DE3) pLysS. For the expression of OsGSTU4 protein, E. coli cells were induced by addition of 1 mM isopropyl \u03b2-D-1-thiogalactopyranoside (IPTG) at 37\u00b0C. Induction of OsGSTU4 protein was analyzed in crude protein extracts by polyacrylamide gel electrophoresis (PAGE). Induced protein was purified using nickel-charged affinity column under native conditions as per manufacturer's protocol .For over-expression in E. coli cells transformed with empty vector (pET28a) and pET28a:OsGSTU4 were cultured overnight at 37\u00b0C and inoculated in fresh Luria-Bertani (LB) medium. After obtaining OD600 of 0.4\u20130.6, the expression of OsGSTU4 protein was induced by adding 1 mM IPTG and bacterial cultures were allowed to grow with or without 1 mM methyl viologen (MV), 1 mM hydrogen peroxide (H2O2), 300 mM mannitol and 300 mM sodium chloride (NaCl). After 3\u20134 h of incubation, growth of cultures was measured using spectrophotometer . For measurement of GST activity, total protein was extracted from bacterial cultures by resuspending the pellet in lysis buffer supplemented with 1 mM phenylmethanesulfonyl fluoride (PMSF) and 2 mg/ml lysozyme followed by sonication. The supernatant was used to measure GST activity in pET28a and pET28a:OsGSTU4 transformed cells under non-stressed and stressed conditions using 1-chloro-2, 4-dinitrobenzene (CDNB) as a substrate, as described earlier psGFPcs:OsGSTU4 construct was made by fusing the complete ORF of OsGSTU4 in psGFPcs vector ApaI and XmaI restriction sites. psGFPcs is a green fluorescent protein (GFP) tag containing vector used in this study to visualize the localization of OsGSTU4. The empty vector (psGFPcs) and fusion construct (psGFPcs:OsGSTU4) were transformed on onion peels by particle bombardment method as described previously For transient expression in onion epidermal cells, a OsGSTU4 was PCR amplified using gene-specific primers (TATTCTAGAATGGCCGGTGGAGGA and TATGGATCCTCAGTTGGTGGCAG) and cloned in pBI121 vector in XbaI and BamHI restriction sites. Further, pBI121:OsGSTU4 construct was confirmed by restriction digestion and sequencing and transformed into Agrobacterium strain GV3101. Agrobacterium-mediated transformation of Arabidopsis plants was performed via floral dip method 4 and T5 generation) as described previously The complete ORF of 2O2) and root growth of WT and transgenics were measured as described earlier 2O2. Percentage of germination was recorded after 5 days as described previously To assay the response of WT and Arabidopsis transgenic seedling roots towards plant hormones, auxin (IAA) and abscisic acid (ABA), and abiotic stresses , seedlings were grown vertically on Petri plates containing MS medium. After 5 days of growth, seedlings were transferred onto fresh MS medium supplemented with various concentrations of plant hormones and abiotic stresses conditions or in the presence of 100 mM and 200 mM NaCl for salinity stress and 5 \u03bcM and 10 \u03bcM MV for oxidative stress as described previously To evaluate tolerance towards salinity and oxidative stresses, WT and transgenic Arabidopsis seedlings were grown in Petri plates. Seedlings with uniform growth for WT and transgenic lines were transplanted into plastic pots after 15 days. Stress tolerance assay was performed on one-month-old Arabidopsis seedlings by irrigating them with 200 mM NaCl for salinity and 25 \u03bcM MV for oxidative stress. Phenotypic and growth parameters, such as plant height (growth), biomass and survival rate were scored after imposing one week of stress treatments.2O2 and superoxide (O\u2022\u20132) levels. The detached leaves from WT and transgenic Arabidopsis plants grown under non-stressed (water), salinity (200 mM NaCl) and oxidative (25 \u03bcM MV) stress conditions, were placed in 1 mg/ml solution (pH 3.8) of 3, 3\u2032-diaminobenzidine (DAB) for detection of H2O2 and nitro blue tetrazolium (NBT) solution for O\u2022\u20132 and incubated at room temperature for 4 h in dark. After staining, accumulation of H2O2 and O\u2022\u20132 were detected as brown color precipitate and blue spots, respectively. The stained leaves were kept in absolute ethanol for 4 h to remove chlorophyll. Subsequently, the bleached leaves were transferred in 70% ethanol and photography was performed.To detect ROS levels in WT and transgenic Arabidopsis plants, we estimated HGST activity in the protein extracts of WT and transgenic plants under non-stressed, salinity and oxidative stress conditions was measured as described above. The estimation was carried out from at least three independent biological replicate samples.UBQ5 and PP2A genes were used as internal control for gene expression data analysis in rice and Arabidopsis, respectively.Total RNA was isolated from transgenic Arabidopsis seedlings by using TRI reagent according to manufacturer's instructions . First strand cDNA was synthesized using 1 \u03bcg total RNA. Real time PCR analysis was performed in three biological replicates as described previously For microarray analysis, transgenic and WT plants were grown for two weeks under normal growth condition and three independent biological replicates were harvested. RNA isolation was done using TRI reagent (Sigma), and quality and quantity of RNA were checked by Agilent Bioanalyzer . Microarray experiment was performed using Affymetrix GeneChip 3\u2032-IVT express kit according to manufacturer's instructions as described previously P-value of \u22640.05 was considered significant.Data of 10\u201312 seedlings for root growth measurement, 40\u201360 seeds for seed germination assay, four to five leaves for leaf disc assays and 10\u201312 plants for phenotypic evaluation from WT and transgenic plants were collected. Each experiment was repeated at least three times. Mean value and standard error (SE) were calculated. Student's t-test was performed to reveal significant differences between WT and transgenic lines. A OsGSTU4, was induced by dehydration, salt and cold stresses. This gene was found to be preferentially expressed in the root and seed development stages and also found to be highly up-regulated by plant hormones, abiotic and biotic stress conditions. Here, we sought to validate the role of OsGSTU4 in various abiotic stress responses. We examined the expression of OsGSTU4 in various tissues/developmental stages , and hormone and abiotic stress treated samples via real-time PCR analysis and oxidative (MV and H2O2) stress conditions as well. The expression analysis of OsGSTU4 in this study corroborated our previous observation In our previous study, we observed differential expression of several GST gene family members in the presence of various plant hormones, and abiotic and biotic stresses; and majority of them belonged to tau class, based on microarray data analysis analysis . We obseOsGSTU4 coding sequence in psGFPcs vector and expressed the fusion protein transiently in onion epidermis cells and Arabidopsis leaf epidermal cells under the control of 35S promoter , osmotic (20%) and oxidative stresses (2O2 (2O2 (11\u201314%) in different transgenic lines. However, we did not find any difference due to osmotic stress in seed germination and root growth under various concentrations of mannitol (data not shown).Further, we analyzed the response of transgenic and WT plants to various abiotic stresses. In root growth assays, we observed increased tolerance in roots of transgenic lines under salinity and oxidative stresses. The root growth of transgenic lines was better under salinity and oxidative stress as compared to WT. The root length of transgenic seedlings was significantly higher in the presence of 150 mM NaCl (11\u201320%), 1 \u03bcM MV (3\u20135%) and 4 mM H(15\u201317%) . In seed7%) , 200 mM NaCl and 25 \u03bcM MV (oxidative stress). There were no phenotypic differences between WT and transgenic lines under control conditions. However, transgenic plants showed tolerant phenotype (remained green and less growth retardation) and were healthier than WT under stress conditions. After treatment, WT plants showed more wilting as compared to transgenic lines. After one week of treatments, phenotypic features (height of the plants (growth), biomass (plant weight) and survival rate) were recorded. These parameters further confirmed the tolerant phenotype of transgenic lines at quantitative level. During salinity stress (200 mM NaCl), we observed tolerant phenotype in transArabidopsis and WT plants. The results showed that GST activity was slightly higher in the transgenics as compared to WT plants and was comparable among the transgenic lines under control conditions levels were also measured in leaves of transgenic and WT plants, respectively, under non-stressed and stressed conditions. The leaves of WT plants showed higher level of H2O2 grown under normal growth condition was performed. The data analysis identified a total of 63 genes (45 up-regulated and 18 down-regulated) significantly differentially expressed in the transgenic line as compared to WT in transgenic rice Suaeda salsa into rice The overexpression of 2O2 and O\u2022\u20132 accumulation in Arabidopsis transgenic plants under stress conditions. Although the transgene mRNA levels varied among the transgenic lines analyzed, the protein (GST) activity was similar under control conditions. We did not observe significant correlation between mRNA level/protein activity and response to hormones and abiotic stresses in the transgenic lines. The GST activity was found to be significantly higher in over-expressing transgenic lines under various stress conditions. The transcript level of putative GST and antioxidant enzyme encoding genes was also higher in the transgenic line. The E. coli cells transformed with OsGSTU4 showed higher GST activity and thus increased tolerance to various abiotic stresses as compared to the cells transformed with empty vector, suggesting the protective role of enhanced OsGSTU4 expression. The exposure of plants to various abiotic stresses leads to oxidative damages OsGSTU4 can eliminate/reduce the level of ROS species under stress conditions. Several reports showed enhanced tolerance to various abiotic stresses, such as salinity, oxidative, osmotic and drought, with increased GST activity due to the over-expression of GSTs 2O2 and O\u2022\u20132 due to the higher antioxidant (GST) activity, at least in part, might directly or indirectly contribute to the enhanced stress tolerance of OsGSTU4 over-expression transgenic Arabidopsis plants.Interestingly, we observed reduced HOsGSTU4, in response to various abiotic stresses. In E. coli, the expression of OsGSTU4 imparts tolerance to various stresses. The transgenic Arabidopsis plants expressing OsGSTU4 also showed better seed germination, root and plant growth and increased capacity to retain chlorophyll content in plant leaves as compared to WT plants under salinity and oxidative stress conditions. The over-expression Arabidopsis transgenic lines showed lower accumulation of ROS and increased GST activity. The increased salinity and oxidative stress tolerance might be related to the higher level of GST activity in transgenic lines due to the overexpression of OsGSTU4. These results suggest that OsGSTU4 can be used as a suitable candidate to improve salinity and oxidative stress tolerance in crop plants.In conclusion, we investigated the role of a rice tau class GST gene, Figure S1Sub-cellular localization of OsGSTU4 in Arabidopsis leaf epidermis. Images were observed in GFP filter by confocal microscopy after 24 h of incubation.(PDF)Click here for additional data file.Figure S2Growth/phenotype of OsGSTU4 over-expression transgenic and WT plants under normal growth condition. (A) One-month-old plants grown in plastic pots in vermiculite. (B) Appearance of WT and transgenic plants in mature stage.(PDF)Click here for additional data file.Figure S3Leaf disc assays showing salinity and oxidative stress tolerance. Effect of salinity (A) and oxidative stress (B) on the leaves of transgenic and WT plants. For stress treatment at least 20 plants and 4\u20135 leaves from each line were used.(PDF)Click here for additional data file.Figure S4Abiotic stress tolerance of OsGSTU4 expressing transgenic lines. Phenotypic characterization of four-week-old transgenic and WT under salinity (A) and oxidative (B) stress. Photographs of plant phenotypes were taken after one week of stress treatment using 200 mM NaCl and 25 \u03bcM MV.(PDF)Click here for additional data file.Table S1List of primers used for real time PCR analysis in this study.(PDF)Click here for additional data file.Table S2List of genes differentially expressed in transgenic line as compared to wild-type.(PDF)Click here for additional data file."} +{"text": "Delphi methods are common in the development of core sets with panels including patients and healthcare professionals (HCPs) as important key stakeholders. Individuals are provided with feedback between survey rounds but methods for considering different stakeholders' views are lacking. This study explored the influence of randomised stakeholder feedback on responses.In a survey to develop a core set for oesophageal cancer surgery, 185 patients and 126 HCPs rated 67 items on a scale of 1: not essential to 9: absolutely essential (Round 1). Feedback on each item was provided in Round 2 with participants randomly allocated to 1) patients receiving own-group feedback, 2) patients receiving both patient and HCP feedback, 3) HCPs receiving both patient and HCP feedback and 4) HCPs receiving own-group feedback. \"Important\" items were defined using a predetermined cut-off (rated 7-9 by >70% of participants). Ratings were summarised as mean scores and differences between randomised groups compared.147 patients and 107 HCPs completed Round 2. Patients receiving only own-group feedback rated more items as important (Group 1: 20 important items versus Group 2: 18 important items). However, HCPs receiving both-group feedback rated more items as important (Group 3: 21 important items versus Group 4: 16 important items). For individual items, the difference in mean scores between randomised groups was small, although there was a trend for HCPs ratings to be influenced by patient feedback.Evidence suggests panel composition and the feedback provided may influence results. Researchers should carefully consider these methodological factors when designing a Delphi study."} +{"text": "Single nucleotide polymorphism (SNP) validation and large-scale genotyping are required to maximize the use of DNA sequence variation and determine the functional relevance of candidate genes for complex stress tolerance traits through genetic association in rice. We used the bead array platform-based Illumina GoldenGate assay to validate and genotype SNPs in a select set of stress-responsive genes to understand their functional relevance and study the population structure in rice.indica accessions compared with other rice sub-populations based on different population genetic parameters. The inferred ancestry of 16% among rice genotypes was derived from admixed populations with the maximum between upland aus and wild Oryza species.Of the 384 putative SNPs assayed, we successfully validated and genotyped 362 (94.3%). Of these 325 (84.6%) showed polymorphism among the 91 rice genotypes examined. Physical distribution, degree of allele sharing, admixtures and introgression, and amino acid replacement of SNPs in 263 abiotic and 62 biotic stress-responsive genes provided clues for identification and targeted mapping of trait-associated genomic regions. We assessed the functional and adaptive significance of validated SNPs in a set of contrasting drought tolerant upland and sensitive lowland rice genotypes by correlating their allelic variation with amino acid sequence alterations in catalytic domains and three-dimensional secondary protein structure encoded by stress-responsive genes. We found a strong genetic association among SNPs in the nine stress-responsive genes with upland and lowland ecological adaptation. Higher nucleotide diversity was observed in SNPs validated in biotic and abiotic stress-responsive rice genes can be used in association analyses to identify candidate genes and develop functional markers for stress tolerance in rice. Single nucleotide polymorphisms (SNPs) represent a robust class of molecular markers . SNP marindica (93\u201311) and japonica (Nipponbare) of these loci were used to classify the 91 rice genotypes into one of three clusters: (i) homozygous AA (japonica \u201cNipponbare\u201d), (ii) homozygous BB (indica \u201c93\u201311\u201d), and (iii) heterozygous AB of 32,942 genotype calls were identified as missing data. The remaining 32,155 yielded successful genotype calls giving a high average call rate of 98% per valid SNP for rice genotypes. When we increased the Gen-Train score cut-off value to \u22650.4, the average missing data rate per successful SNP in rice genotypes decreased to \u22640.5%. Three hundred twenty-five (90%) of the 362 SNP loci, which produced 29,575 genotype calls, showed polymorphism . SNP loci that differentiated any two individual genotypes belonging to two different groups were considered polymorphic between the groups to which they belonged. Based on this criterion, 254 (78%) were polymorphic between O. nivara and O. sativa, 168 (52%) between O. rufipogon and O. sativa, and 28 (9%) between O. rufipogon and O. nivara between O. nivara and indica, 215 (66%) between O. nivara and long-grained aromatics, and 76 (23%) SNPs between O. rufipogon and short-grained aromatics. Within O. sativa, the most polymorphism was observed between indica and japonica and the least was between japonica and short-grained aromatics and lowest between the two japonica genotypes transitions and 118 (36%) transversions, were validated using the \u201crice OPA-1\u201d high-throughput bead array-based assay. The higher frequency of transitions versus transversions in validated SNPs was comparable to the indica and two genotypes of wild species from 30 lowland indica and 19 aromatic rice genotypes. Four of these SNPs \u2014 found in a MYB family transcription factor, a sucrose transporter, a calcium dependent protein kinase and a WRKY family transcription factor and NB-ARC domains (LOC_Os04g30930) differentiated the bacterial leaf blight (BLB) resistant indica rice genotypes Aditya from BLB susceptible aromatic rice genotypes. Another two SNP loci found in genes containing serine threonine protein kinase (LOC_Os09g37949) and leucine-rich repeat (LOC_Os04g19750) domains differentiated eight rice blast resistant indica genotypes from the susceptible indica genotypes followed by transcription factors \u2014 showed amino acid sequence change in DNA binding/signal sensing domains (Table OsWRKY35V2), which was found substituted by serine (S) in the lowland genotypes. This possibly resulted in decreased binding affinities of WRKY domains to invariant \u2018TGAC\u2019 core of W box. It is likely that substitution mutations in WRKY impart differential hydrophobic interactions and beta sheet\u2019s stability thus creating a varied zinc finger and DNA binding protein structure between drought tolerant upland and sensitive lowland rice genotypes compared with long (0.40) and short (0.32) grained aromatics, japonica (0.2841) and aus/wild species (0.25) was evident. We found the average nucleotide diversity (PIC =0.44) across 263 SNP loci in candidate abiotic stress response genes to be higher specifically in upland indica genotypes than that observed in the 62 candidate biotic stress-related genes (PIC = 0.27).The polymorphism information content (PIC) based on SNPs in the stress-related rice genes varied widely across the 89 indica and japonica/aromatic , two wild species O. rufipogon and O. nivara. Molecular genetic variation among and within these four sub-populations (determined by pair-wise estimate of divergence (mean FST) and genetic distance (Dij) based on 325 SNP loci), revealed a wide quantitative genetic differentiation in O. sativa and minimum between aromatics and japonica (0.28) levels revealed that at K value of 2 all genotypes were classified into two distinct sub-populations, r Figure . At thisr Figure . The ricindica (possibly upland aus type) and the wild Oryza species while between japonica and long-grained aromatics it was\u2009~\u20098%. Tripura Medicinal Rice had maximum (54%) admixtures of japonica followed by that of indica (18%), traditional Basmati (15%), short-grained aromatics (5%) and the wild (2%) species followed by chromosomes 7 (59%) and 1 (57%); chromosome 6 contained maximum introgression (68%) of indica. In contrast there was equal sharing (~ 49%) of genomic regions in chromosomes 2 and 8 of japonica and indica. Introgression frequency (based on the number of recombination events) was the most in chromosome 1 showed strong association with ecological adaptation at P < 0.01 and R2 = 1.00.Genetic association analysis using genotyping data of 325 SNPs in stress-responsive genes and phenotypic information of 91 rice genotypes belonging to different ecosystems revealed 18 non-synonymous SNPs associated with ecological adaptation at Illumina GoldenGate platform has shown exceptional performance with regard to throughput, reproducibility, genotype call rate and assay development success rate among several SNP genotyping assays involving human and a few plant species, ,34,43. Iindica genotype 93\u201311 contributing to false SNPs at 37 of 362 loci for which genotype calls were made. With the recent availability of high quality genomic sequence from multiple genotypes , which is expected to be much more informative in plant genomes [The intensity data for each SNP were normalized and cluster positions were assigned using Illumina BeadStudio Genotyping software module. The quality scores represented by GenCall and GenTrain scores were estimated for each SNP call that reflected the degree of separation between homozygous and heterozygous clusters for each SNP locus and placement of individual SNP call for each genotype within a cluster . Minimum genomes . All allThe SNPs in stress-responsive rice genes were categorized according to nucleotide substitutions as either transitions (C/T or G/A) or transversions , and their frequency of occurrence determined individually in different rice genotype groups. The physical position of each validated SNP locus showing allelic variation was determined based on their annotations on the 12 rice chromosomes as described above. The physical locations (bp) of validated SNPs on the 12 chromosomes were used in Graphical GenoTypes Version O. sativa and wild rice genotype groups using the \u201cpreferred\u201d and \u201cunpreferred\u201d concepts of codon usage pattern in Oryza[ab intio three dimensional secondary protein structure and active catalytic domain binding sites with ligands. The high quality protein model of correct topology and protein-ligand complex active binding sites was selected based on high confidence (C\u2265\u22121.5) and binding site (BS\u22650.5) cut-off scores.The divergence of coding sites in each of the variant SNP loci based on derived non-synonymous substitutions (degree of amino acid changes) was analyzed individually for in Oryza. Amino a in Oryza to deter in Oryza,78 for pDNaSp 4.0 [ij) across the genotypes. The genotypic data were used in a model-based program \u201cSTRUCTURE\u201d [indica, aromatic, japonica and aus/wild rice genotype groups at \u03b1 value less than 0.2 was selected. Twenty independent runs with K = 4 were carried out to determine the consistency of results obtained. Various population genetic parameters including fixation of different SNP loci in different sub-populations and their efficiency for detecting genetic variability (FST) and degree of admixture within and between groups were estimated.The SNP loci validated in the stress-responsive gene sequences across 91 diverse rice genotypes were aligned using CLUSTALW multiple sequence alignment tool in MEGA 4.0 and NaSp 4.0 to calcuNaSp 4.0 and geneRUCTURE\u201d to deter2 value \u22650.90 were selected for further analysis.Genotyping data of validated SNP loci in stress-responsive rice genes and phenotypic information of 91 rice genotypes belonging to three different ecosystems namely, upland, medium land and lowland domain (LOC_Os04g19750) showing differentiation between eight known blast resistant indica and 30 blast susceptible indica rice genotypes.Click here for fileOptimization of number of sub-populations varying from K = 2 to K = 5 to determine the best possible population structure for 91 rice genotypes.Click here for filePair-wise estimates of genetic variance (FST) among four O. sativa sub-populations.Click here for fileDomesticated and wild Oryza genotypes used in the study and their inferred ancestry coefficients in population genetic structure analysis.Click here for fileGraphical genotyping using 325 SNP loci validated through Illumina GoldenGate assay across 91 rice genotypes based on their ascending order of physical location (bp) on 12 rice chromosomes giving allele sharing maps of individual rice genotypes.Click here for fileGenomic constitution of rice genotypes based on introgression of indica and japonica alleles on the 12 rice chromosomes.Click here for fileGraphical genotyping of 12 Tripura Medicinal rice chromosomes.Click here for file"} +{"text": "Diabrotica virgifera virgifera Le Conte) provides a novel tool for pest control. Previous studies have shown that RNAi of DvSnf7, an essential cellular component of endosomal sorting complex required for transport (ESCRT), caused deficiencies in protein de-ubiquitination and autophagy, leading to WCR death. Here we investigated the detailed mechanism leading to larval death by analyzing the ultrastructural changes in midgut enterocytes of WCR treated with double-stranded RNA (ds-DvSnf7). The progressive phases of pathological symptoms caused by DvSnf7-RNAi in enterocytes include: 1) the appearance of irregularly shaped macroautophagic complexes consisting of relatively large lysosomes and multi-lamellar bodies, indicative of failure in autolysosome formation; 2) cell sloughing and loss of apical microvilli, and eventually, 3) massive loss of cellular contents indicating loss of membrane integrity. These data suggest that the critical functions of Snf7 in insect midgut cells demonstrated by the ultrastructural changes in DvSnf7 larval enterocytes underlies the conserved essential function of the ESCRT pathway in autophagy and membrane stability in other organisms.The high sensitivity to oral RNA interference (RNAi) of western corn rootworm (WCR, Diabrotica virgifera virgifera (WCR) is the most important insect pest of maize in North America, and recently also has become a problem in Europe Bacillus thuringiensis (Bt) Cry toxins have been widely implemented in North America and have largely replaced the use of chemical insecticides for WCR control Western Corn Rootworm 50 of 4.3 ng dsRNA per mL diet, demonstrating the potential of DvSnf7 as a RNAi target for WCR control.Oral RNA interference (RNAi) is a potential next-generation biotechnology that can be used to suppress a target gene of a pest insect thus offering a new approach for rootworm control D. melanogaster, Snf7 suppression is associated with endosomal cargo sorting The Snf7 protein (Snf7/vps32 in yeast and vps32 in nematodes) belongs to a family of proteins with an evolutionary conserved function and is part of the endosomal sorting complex required for transport-III (ESCRT-III). Multiple functions for Snf7 include: 1) autophagy Targeting Snf7 with ds-DvSnf7 disrupts a vital cellular function that leads to WCR stunting after five days Western corn rootworm (WCR) larvae were obtained from Crop Characteristics and reared at Monsanto Company . Second-instar larvae were fed an artificial diet containing ds-DvSnf7 or ds-GFP (1000 ng dsRNA/mL diet) or without dsRNA following the protocols described 2O, submerged in fixative , incubated under vacuum (620 mm Hg) for 4 hours, and then further fixed overnight (14 hours) without vacuum at room temperature (RT). When sampling specifically for regions in the 3rd to 4th abdominal segments pertaining to the midgut region, an additional incubation with fixative for 24 hours was performed after samples were cut into smaller pieces along their lengths. All fixed samples were washed (3\u00d730 min) and then incubated overnight at 4\u00b0C in 0.1 M cacodylate buffer pH 7.2 (wash buffer) and refixed in 1% osmium tetroxide in 0.1 M cacodylate buffer pH 7.2 . The refixed samples were washed 3 times for 1 hour each and dehydrated in ethanol series (50-100%), 3\u00d720 min each. Samples were stored for 2 days at 4\u00b0C in 100% ethanol. Spurr's low-viscosity resin was used for infiltration and embedding. Dehydrated samples from the posterior larval abdomen were resin infiltrated at RT as follows: 30 min in 100% propylene oxide (PO), 4 hours in PO:resin (2\u22361), overnight (14 hours) in PO:resin (1\u22361), 1 hour in PO:resin (1\u22363), 4 hours in 100% resin under vacuum (680 mm Hg), and an overnight (14 hours) incubation rd or 4th segment and polymerized at 60\u00b0C for 2 days. The ultrathin resin sections (\u223c95 nm) were cut with an ultramicrotome (Reichert Leica) using a diamond knife , transferred to copper grids and contrasted with 2% uranyl acetate . Sections were observed at 80 kV using a Philips CM-100 transmission electron microscope . Images were captured with an AMT digital image capturing system at optimal magnification , and image processing was performed using Adobe Photoshop CS5. In this study, three individuals were used for each treatment.Whole larvae were briefly washed in deionized HPrevious studies have shown that oral RNAi for DvSnf7 in WCR efficiently suppresses the target gene DvSnf7 in a systemic manner Electron microscopy of control/dsGFP-treated enterocytes revealed the presence of regularly shaped and sized endosomes, lysosomes and autolysosomes D.In contrast, structures in macroautophagic complexes were observed in ds-DvSnf7 treated enterocytes, which resemble multi-lamellar bodies [21]. Sivps32 (Snf7 ortholog) mutant Drosophila. Taken together, it is likely that accumulation of macroautophagic complexes due to the lack of fusion with lysosomes and absence of autolysosomes in ds-DvSnf7-treated enterocytes indicates malfunctioning of autophagic processes.The observed absence of autolysosomes in ds-DvSnf7 treated enterocytes in this study corroborates our previous findings showing lack of acidic lysosomal activity in ds-DvSnf7-treated enterocytes of WCR In addition, macroautophagy is a subtype of autophagy, in which the non-specific engulfment of bulk cytoplasm or organelles occurs The progression of pathological symptoms in enterocytes from WCR fed ds-DvSnf7 can be summarized into 4 phases. In the first phase, cell sloughing and apical swelling were observed in midgut enterocytes with the frequent occurrence of macroautophagic complexes . In the One of the most important functions of Snf7 is to recruit the de-ubiquitinating enzyme (DUB) for recycling ubiquitinated proteins Taken together, the major ultrastructural pathological effects in DvSnf7 RNAi enterocytes were macroautophagic complexes that are likely the consequence of the defective fusion between the multi-lamellar bodies and lysosomes. Through EM analysis we were able to visualize the possible essential function of Snf7 in the fusion of autophagic complexes, which is essential for autophagy. The loss of cell membrane integrity and cell-cell junctions points to an impairment in the process of de-ubiquitination of membrane signaling proteins. The severe cellular defects in the enterocytes described in this study are likely the major cause of death at the organismal level after ds-DvSnf7 treatment of WCR. The lethality caused by oral DvSnf7RNAi in WCR suggests malfunctioning of cellular processes such as autophagy and membrane stability."} +{"text": "Knowledge of the effectiveness of cardioprotective medication in women with suspected ischemia is hampered by limited clinical trial data and heterogeneous risk. We assessed the effectiveness of cardioprotective medical treatment in women while adjusting for disease severity determined by cardiovascular magnetic resonance imaging (CMRI).Effectiveness of cardioprotective medication will be evident in women at elevated risk determined by CMRI.Women (n=113), mean age 58\u00b112 years, undergoing coronary angiography for symptoms suggestive of myocardial ischemia additionally underwent myocardial perfusion and cardiac function CMRI evaluation. Previously we identified four ischemic heart disease components associated with adverse cardiac events: 1) coronary artery stenosis > 50%, 2) low global myocardial perfusion, 3) high cardiac energy utilization, and 4) myocardial wall thickness > 10 mm. The latter three disease components were measured using CMRI. Women were stratified into 5 groups based on number of ischemic heart disease components (0-4). Self-reported medication use included angiotensin converting enzyme inhibitors, beta blockers (BB), calcium channel antagonists (CCA) and nitrates. During follow-up (32\u00b117 months) time to first adverse event was analyzed. Cox proportional hazard regression was performed to examine the cardioprotective effectiveness of medications when accounting for ischemic heart disease severity.Medication usage did not differ across ischemic heart disease strata. Ischemic heart disease stratification predicted adverse cardiac events and the medication combination of a BB or CCA and no nitrate were independently associated with reduced risk of events . The model remained significant after adjustment for age and Framingham risk score.Using a combined model of ischemic heart disease severity and medication use, the medication combination of beta-blocker or calcium antagonist and no nitrate were associated with a decreased risk of adverse cardiac events in women. Further investigation of these results should be done with a prospective randomized clinical trial."} +{"text": "We investigated whether human protein C (PC) concentrate to restore physiological values in adult septic shock patients can influence microcirculatory blood flow.n = 18). In both groups, NE was titrated to achieve a MAP between 65 and 75 mmHg. Data from right heart catheterization and sidestream dark-field imaging were obtained at baseline and after 24, 48 and 72 hours.We enrolled 36 septic shock patients with plasma protein C activity <60%. Patients were randomly allocated to be treated with either a continuous infusion of PC concentrate at 3 UI/kg/hour for 72 hours to reach plasma protein C activity between 70 and 120% or a standard treatment and perfused vessel density (PVD). Results are summarized in Table The administration of human PC concentrate did not influence microcirculatory blood flow in septic shock patients."} +{"text": "To evaluate the ability of an inexperienced observer (IO) to reliably assess mid-wall late enhancement (MLE) and to assess the prevalence of MLE in patients with various cardiac diseases.Late gadolinium enhancement (LGE) in cardiac MRI (cMRI) has been described as a valid tool to discriminate between cardiac diseases. It has been postulated that MLE especially occurs in patients with dilated cardiomyopathy (DCM) and myocarditis with a prognostic impact in these patients. Nevertheless, it can be difficult to differentiate true MLE from common artifacts as motion blur, partial volume effects (PVE) or wrong inversion times (TI), especially for the IO.We examined 97 consecutive patients , which were referred to our department for a cMRI for various clinical indications (37 ischemic heart diseases (ICM), 16 myocarditis, 5 DCMs, 2 restrictive cardiomyopathies (RCM), 5 hypertrophic obstructive or non-obstructive cardiomyopathies, 8 congenital heart diseases (CHD), 12 patients with arrhythmias and 12 others. Besides Cine-sequences, standard LGE-sequences (IR-GRE) and phase sensitive inversion recovery (PSIR) sequences were applied and evaluated by two independent blinded observers (1 inexperienced observer (IO) with 2 months of cMRI experience and 1 experienced observer (EO) (3 years of experience). The results of the EO (Table The IO described suspected MLE in 43/97 patients (44%), which were false positive in 28/43 (65%). Only 18/97 (19%) were true MLE. Reasons for false positives were wrong TI in 39% (Table MLE is a common finding not only in patients with DCM and myocarditis, but also in patients with ICM, RCM, CHD or patients with different arrhythmias without an underlying structural heart disease. Standardized criteria for the detection/definition of MLE are mandatory to reduce the number of false positive results, which can be higher than 50%, especially when cMRI is interpreted by an inexperienced cardiac MRI user."} +{"text": "Brassica napus\u2005L. from a trace elements contaminated field and a non-contaminated control field were characterized genotypically and phenotypically. Correspondence analysis of the genotypic data revealed a correlation between soil and rhizosphere communities isolated from the same field, indicating that local conditions play a more important role in influencing the composition of (rhizosphere) soil bacterial communities than root exudates. In contrast, endophytic communities of roots showed a correlation between fields, suggesting that plants on the two fields contain similar obligate endophytes derived from a common seed endophytic community and/or can select bacteria from the rhizosphere. The latter seemed not very likely since, despite the presence of several potential endophytic taxa in the rhizosphere, no significant correlation was found between root and rhizosphere communities. The majority of Cd/Zn tolerant strains capable of phosphorus solubilization, nitrogen fixation, indole-3-acetic acid production and showing 1-aminocyclopropane-1-carboxylate deaminase capacity were found in the rhizosphere and roots of plants growing on the contaminated field.Cultivable bacterial strains associated with field-grown Due to atmospheric deposition from four zinc ore smelters in the Dutch-Belgian border region, soils in the Campine region got enriched with cadmium (Cd), zinc (Zn) and lead (Pb) (Vangronsveld Brassica napus L. (rapeseed) as a candidate phytoextraction crop because it combines high biomass production with a good tolerance to Cd and Zn ] and a non-contaminated field in Alken . The main objectives of this study were to extend our knowledge on the poorly known bacterial communities associated with B.\u2009napus and to identify PGP, Cd tolerant and Cd solubilizing bacteria which might increase biomass production and Cd uptake by rapeseed growing under the unfavourable environmental conditions occurring on contaminated fields.Since the natural habitat is considered as an interesting model for the evolution of TE tolerant PGP microorganisms and a contaminated field (TE-F) Table\u2009.For both fields, the number of cultivable strains recovered from the bulk soil and roots were significantly lower compared with the rhizosphere soil. Significantly more cultivable strains were isolated from the bulk soil compared with the roots at the contaminated field. Considering compartments, no differences in the amount of isolated strains between fields were observed. At both fields, the number of different bacterial genera was higher in the rhizosphere soil than in bulk soil and root.B.\u2009napus were characterized by amplified 16S rDNA restriction analysis (ARDRA) using the restriction enzyme HpyCH4IV. One representative member of all strains with identical fingerprints was sequenced for identification by means of Sequence Match at the Ribosomal Database Project II. All (except Chryseobacterium DQ337589) strains have a sequence match score higher than 0.900, which indicates a confident identification to the genus level (Appendix\u2009S1). Also the neighbour-joining tree clustered strains belonging to the same genus together, confirming the results of the 16S rRNA genes-based identification procedure (Appendix\u2009S2). The identification resulted in 37 different bacterial genera recovered from the contaminated field and 29 from the control field , Bacillus (38.7%) and Leifsonia (5.8%) and at the contaminated field by Arthrobacter (26.9%), Leifsonia (15.7%), Staphylococcus (10.0%), Burkholderia (7.4%), Variovorax (6.4%) and Rhodopseudomonas (5.6%). The major part of the cultivable rhizosphere bacteria at the control field consisted of Bacillus (43.2%), Pseudomonas (19.3%), Variovorax (13.7%) and Staphylococcus (8.3%) and at the contaminated field of Leifsonia (29.3%), Staphylococcus (22.7%), Plantibacter (12.3%), Serratia (6.9%) and Microbacterium (6.0%). Variovorax (34.5%), Pseudomonas (29.4%), Bacillus (12.4%), Caulobacter (8.6%) and Pantoea (6.5%) dominated the cultivable root endophytes at the control field, while at the contaminated field it were Pseudomonas (33.0%), Rhizobium (16.1%), Caulobacter (14.9%), Pedobacter (14.7%) and Variovorax (7.8%) , Burkholderia (FJ786047) and Leifsonia (AB278552); in the rhizosphere it were Agrobacterium (GQ428123), Arthrobacter (AB288059), Bacillus (AB188212), Leifsonia (AB278552), Lysinibacillus (AY907676), Pseudomonas (AB369347), Rhodococcus (DQ060386), Sphingobacterium (AJ438176), Staphylococcus (GQ222398) and Variovorax ; and in the roots it were Caulobacter (DQ337549), Labrys (DQ337554), Mycobacterium (FJ719354), Pantoea (EU598802), Pedobacter (GU385862), Plantibacter (AM396918), Pseudomonas , Rhizobium (DQ337581) and Variovorax (GQ861460).The number of genotypically different bacterial strains occurring in the same compartment at both fields (see intersections) increases from bulk soil to rhizosphere soil and roots Fig.\u2009. BacteriMycobacterium (FJ719354), Pedobacter (GU385862), Pseudomonas (FN377713) and Variovorax (GQ861460) were exclusively found in the roots. Pantoea (EU598802), Plantibacter (AM396918) and Rhizobium (DQ337581) were isolated from roots at both fields; at the control field they were not found in bulk neither rhizosphere soil, like at the contaminated field Caulobacter (DQ337549), Labrys (DQ337554) and Pseudomonas (FJ772042) did not occur in other compartments. Other root endophytes were also found in bulk and/or rhizosphere soil . Bacterial strains restricted to the bulk and rhizosphere soil at the control field were Bacillus (GU321095) and Leifsonia (AB278552) and at the contaminated field Agrobacterium (GQ428123), Arthrobacter (AB288059), Bacillus (AB188212), Brevundimonas (EF088675), Methylobacterium (AB220076) and Variovorax (EF419341).Bacterial strains exclusively occurring in one of the investigated compartments at the control respectively the contaminated field are underlined in Fig.\u2009Based on the correspondence analysis (CA) of these data Fig.\u2009, we can anova) test indicated a significant field effect especially when considering tolerance to the highest Cd and Zn concentrations .All purified isolates were screened for their TE tolerance Table\u2009A. PercenAll strains were screened for their potential PGP characteristics Table\u2009B. The peanova test indicated a significant field effect considering phosphorus solubilization, nitrogen fixation and siderophore and acetoin production .The highest percentages of strains able of solubilizing phosphorus, fixating nitrogen and producing siderophores, ACC deaminase and IAA at the control field were found in the roots, while proportionally the most organic acid and acetoin producing strains were detected in the rhizosphere soil. At the contaminated field, percentages of siderophore and ACC deaminase producing strains were highest in the bulk soil, while percentages of phosphorus solubilizing, nitrogen fixating and organic acid/acetoin producing strains were proportionally highest in the rhizosphere soil. The most IAA producing strains at the contaminated field were found inside the roots. The two-way 3)2 selective extraction.Total TE concentrations were determined in bulk soil and plant parts Table\u2009. In addi3)2-extractable Cd, Zn and Pb in the bulk soil were significantly higher at the contaminated field as compared with the control field, like also the Cd and Zn concentrations measured in the roots, stems, leaves and seeds. Lead concentrations in plant parts were below detection limit while total and Ca(NO3)2-extractable Pb concentrations in the soil at the contaminated field were significantly higher than at the control field. Plants grown at the contaminated field contained significantly more Cd and Zn in their leaves compared with the roots and stems, their seeds accumulated acceptable Cd concentrations (<\u20091.14\u2009mg Cd kg\u22121) according to the European standards for animal feed (Appendix\u2009S3). At both fields, plants contained adequate tissue levels of Ca, K, Mg, Na and Zn (data not shown). The Cu and Fe concentrations in plant tissues were just below the prescribed levels for adequate growth but are not expected to be limiting. Total bulk soil Cu and Fe concentrations were significantly higher at the contaminated field whereas the Ca(NO3)2-extractable concentrations were similar at both fields. When plant parts were compared between fields, total Fe concentrations were significantly higher in roots from the control and seeds from the contaminated field.Amounts of both total and Ca according to the European standards for animal feed, unlike the other plant parts harvested on the contaminated field and a contaminated field as well as the characteristics of the isolated bacterial communities that might contribute to improve biomass production and TE uptake and translocation. Approximately 500 morphologically different bacterial strains were isolated from bulk soil, rhizosphere soil and roots of B.\u2009napus at both fields and identified based on 16S rDNA sequencing profiles, a tentative identification method and roots of field-grown B.\u2009napus. Additionally, Kaiser and colleagues , Pedobacter (GU385862), Pseudomonas (FN377713) and Variovorax (GQ861460) Fig.\u2009 who menterium FJ79354, Pedet\u2009al., et\u2009al., All isolated strains were tested for their Cd and Zn tolerance. The highest numbers of strains tolerant to 1.6\u2009mM Cd and 2.5\u2009mM Zn originated from the contaminated field Table\u2009, caused 3)2-extractable Fe concentrations in the soils from both fields were not significantly different (Table\u2009\u22121) was noticed in all parts of plants from the contaminated field; this might inhibit chlorophyll synthesis and chloroplast development and increase ethylene production in plant tissues, eventually leading to decreased remediation efficiency as well as by their host plant (i.e. selection from the rhizosphere/bulk soil and present seed endophytes). The environmental conditions at the contaminated field seem inductive for the occurrence of rapeseed-associated bacteria with potential to enhance Cd phytoextraction. Enriching the rhizosphere with these PGP, siderophore and/or organic acid producing bacteria might enhance TE uptake while endophytes equipped with a TE sequestration system might reduce Cd phytotoxicity. In future inoculation experiments, the B.\u2009napus, soils and plants were sampled after the flowering stage (June 2010). Sampling was performed on a TE -contaminated former maize field in Lommel . Polymerase chain reaction (PCR) amplification of the 16S rRNA genes was performed on aliquots of the extracted DNA using the universal primers, 16S-prokaryotic-R (5\u2032-ACGGGCGGTGTGTRC-3\u2032) and 16S-prokaryotic-F (5\u2032-AGAGTTTGATCCTGGCTCAG-3\u2032) as described previously by Weyens and colleagues .For amplified 16S rDNA restriction analysis (ARDRA), 20\u2009\u03bcl of the PCR products were digested with the HpyCH4IV enzyme and visualized by gel electrophoresis as described by Weyens and colleagues . Bacteriet\u2009al., 4. Strains, not able to grow in the test media (pH\u20097) during incubation [5 (liquid media) to 7 days (solid media) at 30\u00b0C], were considered as not detectable (nd). Media without cell suspension were used as controls.All purified bacterial isolates were screened for TE tolerance (Cd and Zn) and potential PGP characteristics . Before screening, strains were grown in 869 medium (Mergeay 4) or 0.0, 0.6, 1 and 2.5\u2009mM Zn (ZnSO4), tolerance was rated visually medium containing per litre 17\u2009g of MRVP medium (Sigma-Aldrich). After 48\u2009h of incubation, a colorimetric reaction was induced according to Romick and Fleming 2 extraction , a principal component analysis related ordination technique based on chi-square distances, illustrating correlations between compartments. A logit transformation was performed on the proportional data gathered during the phenotypic analyses before analysing them using a two-way anova and post hoc multiple comparison testing (Tukey Kramer). The same method (without the logit transformation) was used to test the bacterial amounts isolated from the different compartments at both fields. TE concentrations were statistically compared between both fields using one-way anova. Transformations were applied when necessary to approximate normality and/or homoscedasticity. In case normality could not be reached, data were analysed using Kruskal\u2013Wallis multiple comparisons test.Percentages of genotypic and phenotypic different strains per mixed sample and their mean percentages per compartment were calculated but not appropriate for"} +{"text": "Dear Editor,Benign Prostatic Hyperplasia (BPH) is a common disease observed in 90 percent of men over 60 . Benign"} +{"text": "Lymph node metastasis is a key event in the progression of breast cancer. Therefore it is important to understand the underlying mechanisms which facilitate regional lymph node metastatic progression.EED and EZH2 in lymph node metastatic tumor cells over primary tumor cells which were validated through real-time PCR. Additionally, immunohistochemical (IHC) staining and quantitative image analysis of whole tissue sections showed a significant increase of EZH2 expressing tumor cells in lymph nodes over paired primary breast tumors, which strongly correlated with tumor cell proliferation in situ. We further explored the mechanisms of PRC2 gene up-regulation in metastatic tumor cells and found up-regulation of E2F genes, MYC targets and down-regulation of tumor suppressor gene E-cadherin targets in lymph node metastasis through GSEA analyses. Using IHC, the expression of potential EZH2 target, E-cadherin was examined in paired primary/lymph node samples and was found to be significantly decreased in lymph node metastases over paired primary tumors.We performed gene expression profiling of purified tumor cells from human breast tumor and lymph node metastasis. By microarray network analysis, we found an increased expression of polycomb repression complex 2 (PRC2) core subunits in situ. Concurrently, PRC2 target protein E-cadherin was significant decreased in lymph node metastases, suggesting PRC2 promotes epithelial mesenchymal transition (EMT) in lymph node metastatic process through repression of E-cadherin. These results indicate that epigenetic regulation mediated by PRC2 proteins may provide additional advantage for the outgrowth of metastatic tumor cells in lymph nodes. This opens up epigenetic drug development possibilities for the treatment and prevention of lymph node metastasis in breast cancer.This study identified an over expression of the epigenetic silencing complex PRC2/EED-EZH2 in breast cancer lymph node metastasis as compared to primary tumor and its positive association with tumor cell proliferation Axillary lymph node metastasis is the single most important prognostic factor for patient survival in situ. The PRC2 complex belongs to polycomb-group (PcG) proteins and functions as epigenetic repressors which silence specific sets of genes through its histone 3 lysine 27 (H3K27) methyltransferase activity In this study, we performed gene expression profiling of freshly isolated tumor cells from primary breast tumor tissues and tumor positive lymph nodes with the goal of identifying therapeutic targets for the treatment of lymph node metastasis. We undertook an integrative approach by combining gene set and network based microarray analyses with validations through real-time PCR and quantitative image analysis of whole tissue sections using histological methods. We found increased expression of polycomb repression complex 2 (PRC2) genes in lymph node metastatic tumor cells compared to primary tumor cells. Additionally, over expression of EZH2, the catalytic unit of PRC2, in lymph node metastasis strongly correlated with tumor cell proliferation To capture subtle yet biologically important changes, the Gene eXpression Network Analysis (GXNA) program To validate our findings, we examined whether the LN Met Set identified in this study could distinguish metastasis from primary tumor in previously published breast cancer microarray datasets using Gene Set Enrichment Analysis (GSEA) http://string-db.org/) EZH2, EED, SUZ12, RBBP4 and their interacting neighbors formed a prominent tightly connected protein network. The relative expressions of these genes are presented as a heat map in EZH2, EED, RBBP4) were found to be significantly up-regulated in lymph node metastasis using the program GXNA. Although not identified through GXNA analyses, E2F3 was up-regulated in 5 out of 6 lymph node metastasis and clustered with EZH2 and EED. However, expression of E2F1 and E2F2 were relatively low in all samples, possibly due to insensitive probes on the microarrays. Two additional PRC2 genes, SUZ12 and RBBP4, were up-regulated in 4 out of 6 metastatic lymph node samples and co-expressed with HDAC1-2. Overall, the network analysis of LN Met Set highlighted up-regulations of the PRC2 complex and the pRB-E2F growth control pathway in lymph node metastasis.To characterize the LN Met Set, we generated a protein-protein interaction network using the STRING database (EZH2 and its binding partner EED in purified tumor cells from 8 paired primary tumor and lymph node samples (EZH2 in metastatic tumor cells compared to primary tumor cells (p\u200a=\u200a0.007); EED demonstrated an increased trend in 6 out of 8 paired samples (p\u200a=\u200a0.054). Real-time PCR analyses of RBBP4 was performed in 6 paired primary tumor and lymph node samples . The C2 collection includes 3272 gene sets collected from various sources such as online pathway databases, publications in PubMed, and knowledge of domain experts.To further explore the mechanisms of PRC2 gene up-regulation in lymph node metastasis, GSEA was used to investigate enrichment patterns of prior defined gene sets in lymph node metastasis and primary tumor. We used gene set C2 from GSEA Molecular Signatures Database version 3.0 were significantly down-regulated in lymph node metastasis, although CDH1 itself did not show differences at the transcriptional level. Lymph node metastasis also over expressed genes associated with resistance to doxorubicin, a cancer chemotherapy drug. Concurrently, lymph node metastasis down-regulated gene sets associated with drug-responses, including cisplatin, a cancer chemotherapy drug and MP470, a novel c-Kit/AXL kinase inhibitor. Collectively, the transcriptional scheme of lymph node metastatic cells exhibited a highly proliferative and aggressive phenotype.GSEA analysis of the gene sets C2 showed that 18 gene sets were up-regulated in lymph node metastasis and 19 gene sets were up-regulated in primary tumor . ComparePreviously it has been shown that EZH2 mediates transcriptional silencing of E-cadherin by trimethylation of H3K27 Metastatic tumor cells arise within the primary tumor cell population and acquire additional genetic and epigenetic changes to enable metastatic outgrowth to the draining lymph nodes and distant organs. Microarray analysis has been widely used to study genetic changes implicated in tumor initiation, progression and metastasis : EZH2, EED and SUZ12. EZH2 is the catalytically active unit of the PRC2 complex which participates in the transcriptional repression of target genes by trimethylation of H3-K27, providing an epigenetic mark for the PRC1 complex binding In vitro studies suggested that EZH2 drives neoplastic transformation of breast epithelial cells by promoting anchorage-independent and invasive growth MYC and E2F through microarray analyses. Bracken et al. previously showed that expressions of the two PRC2 core subunits EZH2 and EED were tightly controlled by the E2F transcription factors. EZH2 and EED are required for cell growth to confer a proliferative advantage and act as essential downstream mediators of E2F function EED-EZH2 genes in LN metastasis, our microarray data showed an increase of E2F3 in 5 out of 6 lymph node metastasis compared to primary tumor. As such, PRC2/EED-EZH2 components could act downstream of E2F genes to provide additional growth advantage for LN metastatic tumor cells. Additionally, MYC may also be responsible for the up-regulation of the PRC2 genes through regulation of microRNA expressions in lymph node metastases. MYC not only stimulates EZH2 expression by repression of its negative regulator miR-26a By profiling purified tumor cells from human breast tumor and lymph nodes, we discovered an up-regulation of epigenetic silencing complex PRC2 core subunits (EED-EZH2) in lymph node metastatic tumor cells compared to primary tumor cells. The PRC2 complex functions as epigenetic repressors by silencing specific sets of genes through its histone 3 lysine 27 (H3K27) methyltransferase activity Furthermore, we found that lymph node metastatic tumor cells showed down-regulation of genes activated by tumor suppressor gene E-cadherin through GSEA enrichment analyses. Further validation of E-cadherin expression through IHC revealed a significant decrease in E-cadherin expressing tumor cells in LN metastasis compared to paired primary tumor, which coincided with an up-regulation of EZH2 in LN metastasis. E-cadherin down-regulation is one of the hallmarks of epithelial-mesenchymal transition (EMT), which is essential for tumor invasion and metastasis Previous microarray studies have focused on potential genetic targets for treating breast cancer metastasis. Our findings showed an over expression of epigenetic silencing complex PRC2/EED-EZH2 in breast cancer lymph node metastasis compared to primary tumor, suggesting epigenetic regulations mediated by PRC2 proteins may provide additional advantage for the outgrowth of lymph node metastasis. In the lymph node metastatic process, PRC2/EED-EZH2 might act downstream of E2F genes and promote EMT through repression of E-cadherin. Epigenetic regulation represents a key, yet poorly understood mechanism contributing to the metastatic process. Future studies are warranted to identify critical epigenetic regulatory mechanisms involved in the metastatic process. These findings provide novel insights into potential therapeutic strategies using epigenetic agents for the treatment/prevention of metastatic disease.Written informed consent was obtained from all patients, and the study was approved by Stanford University\u2019s Institutional Review Board.A total of 20 primary breast tumors and 9 metastatic lymph nodes (LNs) were obtained from breast cancer patients at Stanford Hospital. The patient and tumor characteristics are shown in +CD45\u2212CD140\u03b2\u2212 cells were sorted using FACSAria (BD Bioscience) with gating strategies shown in Fresh breast tumors and tumor positive lymph nodes were minced and enzymatically dissociated with 200 units/ml type III Collagenase and 10 Kunitz units/ml DNase I (Sigma) for 30 minutes to 1 hour at 37\u00b0C. The digestion process was stopped by addition of M199 media containing 10% FBS. Single cell suspensions were generated by filtering cells through a 70 micron cell strainer followed by a 40 micron cell strainer (BD Biosciences). Cells were stained with pan-leukocyte marker CD45 PE-Cy7, fibroblast marker CD140\u03b2 PE (both BD Biosciences), epithelial specific antigen ESA FITC (Biolegend), and a dead cell exclusion marker ViViD (Invitrogen). ESATotal RNA was extracted using the TRIzoL method and amplified using TrueLabeling-PicoAMP\u2122 kit (QIAGEN), followed by Cy3/Cy5 labeling . Cy-labeled patient samples were mixed with the same amount of reverse color Cy-labeled universal human reference (UHR) cRNA (Stratagene Corp.) and hybridized to Agilent's Whole Human Genome Microarray 4\u00d744 K. Image files were generated from microarray slides using Agilent Microarray Scanner G2505B. Quantile normalization and LOESS normalization were used to normalize the raw data. All arrays were calibrated to the same scale and variance stabilizing transformation was applied to the data. After normalization, the generalized log ratio values for each gene were taken to obtain a relative expression level in samples over the UHR. Array data are MIAME compliant and available at National Center for Biotechnology Information\u2019s Gene Expression Omnibus (GSE30480). Additional nonspecific filtering was performed using R-package and 8386 genes were used for the analyses. Briefly, multiple probes for the same genes were averaged when they were highly correlated; otherwise they were left as is. Probes without entrez ID or Gene Ontology annotation were removed. Variance filtering was used to remove genes which showed little or no variability. Average linkage was performed using Cluster 3.0 and heat maps were generated using JavaTreeview. The STRING database was used to generate protein-protein networks GXNA was employed for finding significantly changed networks/subnetworks GSEA was used to determine whether the members of a given gene set were randomly distributed throughout the differentially expressed genes between two phenotypes or associated with a particular phenotype T method. The relative expression of the target gene was expressed as a fold change over UHR. Two-sided paired-Wilcoxon signed rank test was used to assess the statistical significance of differential gene expressions. P-values <0.05 were considered statistically significant.Total RNA was reverse transcribed using sensiscript reverse transcriptase (QIAGEN). Real-time PCR reactions were performed in duplicates with iQ\u2122 SYBR\u00ae Green Supermix (Bio-Rad Laboratories) and specific PCR primers using thImmunohistochemistry was performed on formalin fixed paraffin-embedded tissue sections using a biotin-free polymer detection system. Prior to staining, tissue sections were deparaffinized and antigens retrieved with Diva Decloaker\u2122 in the Decloaking Chamber. A three step staining procedure was performed on tissue sections for the detection of 1) EZH2, proliferation marker Ki67 and pan cytokeratin marker AE1/AE3 and a two step staining procedure was used for the detection of 2) E-cadherin and pan cytokeratin marker AE1/AE3. For the former, tissue sections were incubated with rabbit monoclonal antibody Ki67, followed by incubations with anti-rabbit ALP secondary antibody and color developed by chromogen Vulcan Fast Red. Endogenous peroxidase was blocked by incubation in PEROXIDAZED 1 for 10 minutes and mouse monoclonal antibody EZH2 was applied to the tissue sections followed by anti-mouse HRP secondary antibody. The color for EZH2 was developed by incubating with chromogen DAB. A denaturing solution was applied to denature bound antibodies before staining with the pan cytokeratin antibody , followed by anti-mouse ALP secondary antibody and color was developed with chromogen Ferangi Blue. For the latter, tissue sections were blocked with PEROXIDAZED 1, incubated with mouse monoclonal primary antibody E-cadherin followed by anti-mouse ALP and color developed by Vulcan Fast Red. A denaturing solution was applied before staining with AE1/AE3, followed by anti-mouse ALP secondary antibody and color was developed with chromogen Ferangi Blue. All tissue sections were counterstained in CAT Haematoxylin and mounted. All antibodies and reagents were purchased from Biocare Medical .Whole tissue section images were acquired at 200\u00d7magnification using the imaging system Vectra\u2122 Figure S1GSEA enrichment results of LN Met Set in breast cancer microarray dataset GSE2741. A: Lymph node metastasis vs Primary Tumor. B: Lymph node metastasis vs Normal.(TIF)Click here for additional data file.Figure S2Validation of RBBP4 mRNA expressions by real-time PCR. White bars indicate primary tumor and the adjacent black bars indicate their matched lymph node metastasis.(TIF)Click here for additional data file.Figure S3FACS plots indicating sorting gates for isolation of tumor cells (ESA+CD45-CD140\u03b2-ViVID-). First, fibroblasts and dead cells were excluded by gating CD140\u03b2-ViVID- cells on a CD140\u03b2 versus ViVID plot. Within the CD140\u03b2- live cells gate, a further gate was set on a CD45 versus ESA plot to exclude immune cells and identify ESA+ cells as indicated in the plots. Left Panel: tumor tissue; Right Panel: lymph node metastasis.(TIF)Click here for additional data file.Figure S4Decomposition of multiple chromogen stained histological sections. Vectra is able to decompose the original image into individual stains according to the corresponding spectrum for each chromogen. Pseudocolor can be assigned for each chromogen for better visualization. A: 200\u00d7 original image. B: pseudo-colored image of blue membrane staining of cytokeratin, green nuclear staining of EZH2 and red nuclear staining of Ki67. C: pseudo-colored image of blue membrane staining of cytokerain and green nuclear staining of EZH2. D: pseudo-colored image of blue membrane staining of cytokerain and red nuclear staining of Ki67.(TIF)Click here for additional data file.Table S1Gene sets (C2) differentially expressed in lymph node metastasis vs primary tumor.(XLS)Click here for additional data file.Table S2Patient and Tumor Characteristics.(XLS)Click here for additional data file.Table S3PCR primer pairs.(XLS)Click here for additional data file."} +{"text": "Acute kidney injury (AKI) is a serious complication associated with increased morbidity and mortality in pediatric patients undergoing surgery for congenital heart disease. The aim of this study was to evaluate children with AKI after pediatric cardiac surgery using pediatric-modified RIFLE (pRIFLE) criteria and to investigate the value of serum cystatin C in patients with AKI.Eighty-one children undergoing cardiopulmonary bypass (CPB) for surgical correction of acyanotic congenital heart disease were prospectively enrolled in the study. Serial blood samples were collected to measure serum cystatin C and creatinine levels. The primary outcome measure was AKI, defined as \u226550% increase in serum creatinine from baseline.P = 0.002). No differences were noted with respect to CPB and aortic cross-clamp durations in those with and without AKI (P > 0.05). Postoperative 24-hour inotrope scores were significantly higher in children who developed AKI (P = 0.003). Serum cystatin C concentrations were significantly increased in AKI patients at 2 hours after CPB (P = 0.029) and remained elevated at 24 hours (P < 0.001) and 48 hours (P = 0.001). There was a significant positive correlation between presence of AKI and serum cystatin C levels (P < 0.05). A significant negative correlation was found between age and AKI .Twenty-one children (26%) developed AKI, in which risk occurred in 12 (15%), injury in three (4%) and failure in six (7%) of the patients diagnosed with serum creatinine. Patients with AKI were significantly younger than patients without AKI (AKI develops in 26% of patients after pediatric cardiac surgery. Our results suggest that patients with AKI were younger and had postoperative higher serum cystatin C levels and higher inotrope scores when compared with patients without AKI."} +{"text": "To the Editor: In 2010, detection of henipavirus (Hendra or Nipah virus) and rubulavirus (Tioman or Menangle virus) antibodies in fruit bats in Papua New Guinea (PNG) was reported , New Britain (1997), and Lae (1999 ) . The 20 Of the 20 samples from Madang, 2 (10%) reacted in the Hendra virus ELISA. Of the 147 samples from New Britain and Lae that yielded definitive VNT results, 11 (7.5%) yielded neutralizing antibodies to Hendra virus and 5 (3.4%) to Nipah virus. All samples with antibodies against Nipah virus also had antibodies against Hendra virus; titers against Hendra virus were greater (4 samples) or equivalent (1 sample) to those against Nipah virus. Reciprocal titers against Hendra virus were 5\u2013160 (median\u00a010) and against Nipah virus, 5\u201380 (median\u00a010). None of the 103 samples from Lae had antibodies against Menangle virus .Pteropus conspicillatus) from proximate locations (Lae and Madang), the significant differences in antibody prevalence remained: 0 (95% CI 0\u201323%) in 1999 and 65% (95% CI 50%\u201378%) in 2009.The common and contrasting findings between the study of Breed et al. that had a neutralizing antibody titer to Nipah virus had a higher neutralizing titer to Hendra virus , suggesting that the circulating henipavirus was more similar to Hendra virus than to Nipah virus. These findings are supported at a regional level by those reported for nearby Indonesian islands by Sendow et al. associated with negative ecologic effects (More robust interpretation of the serologic findings of both studies is constrained by the lack of henipavirus sequence data from PNG and neighboring countries. We concur with Breed et al. ("} +{"text": "Smad ubiquitination regulatory factor 2 (Smurf2).A novel, cancer-fighting function was recently discovered for Smad ubiquitination regulatory factor 2, which is an E3 ubiquitin ligase involved in the signaling control of the TGF-\u03b2 and BMP super-family of poly-peptide growth factors [A new guardian of genomic stability against cancerous mutations has been discovered, or so reported in a recent article published in Nature Medicine from Dr. Ying E. Zhang's laboratory at the National Cancer Institute in Bethesda, Maryland . This neSmurf2 and its related protein Smurf1 are members of HECT domain containing ubiquitin E3 ligases. Both of them play pleiotropic roles in many aspects of cellular functions ranging from regulating planar cell polarity during embryonic development to osteogenic differentiation in adult bone formation . HoweverThe authors declare that they have no competing interests.KZ and YBS wrote the manuscript. All authors read and approved the final manuscript."} +{"text": "Post partum cardiomyopathy (PPCM) can occur between the last trimester of pregnancy or within 5 months post partum, in the absence of any history of heart disease. Although rare, it may become unresponsive to medical treatment and can in such cases, require mechanical support or even a heart transplant. Predicting its evolution is unfortunately not possible.Late gadolinium enhancement CMR (LGE) can identify myocardial fibrosis and has shown its prognostic significance in DCM. However, the role of LGE in PPCM as a predictor of outcome has not been reported.The purpose of this study was to assess whether LGE imaging provides prognostic information in the outcome in post partum cardiomyopathy.From 2003 to 2012, 36 women (mean age 34\u00b110 years) diagnosed as PPCM underwent standard CMR with LGE following Gd-DTPA injection at the time of PPCM suspicion and after a mean follow-up period of 33 \u00b120 months for 21 of them (58%).At the time of diagnosis: All patients showed increased left ventricular (LV) volumes , 17 patients showing reduced LVEF (mean LVEF 47.8% \u00b114%) and 15 patients had additionally dilated RV . Only 7 patients displayed additionally reduced right ventricular (RV) function (mean RVEF 54.85\u00b113.5%).CMR data analysis from all patients in Table Initial study showed mid wall LGE in 6 patients (30%) indicating myocardial fibrosis. Among them, one patient had a follow-up scan displaying persistence of myocardial enhancement but improvement in LV volumes and ejection fraction (EF). Four patients out of 6 had dilated LV with reduced LVEF; one showed increased volumes but normal EF and 1 had normal volumes with impaired EF. The RV appeared dilated with impaired EF in 3 patients at baseline (3/6 pts) without improvement on the follow up scan.All the remaining patients without LGE (30/36 pts) had initially a dilated LV, with associated reduced LVEF in 13 pts. Among them, 3 had a complete normalization of their LV volumes and 4 patients had normal LVEF at follow up. RV volumes and RVEF did not change significantly at follow up in this subgroup.None of the patients showed active myocardial inflammation or oedema on STIR images.In our cohort, the presence or absence of myocardial late gadolinium enhancement was not predictive of LV functional recovery in PPCM suggesting that mechanisms other than myocardial fibrosis are involved in the genesis of ventricular dysfunction. None of our patients displayed myocardial edema. However, follow-up results are still awaited.None"} +{"text": "Segmental duplications (SDs) on 22q11.2 (LCR22), serve as substrates for meiotic non-allelic homologous recombination (NAHR) events resulting in several clinically significant genomic disorders.Alu SINEs, are associated much more frequently with duplicated sequences that have undergone active expansion, confirming their role in mediating recombination events. Many copy number variations (CNVs) exist on 22q11.2, some flanked by SDs. Interestingly, two chromosome breakpoints for 13 CNVs (mean length 65 kb) are located in paralogous subunits, providing direct evidence that SD subunits could contribute to CNV formation. Sequence analysis of PACs or BACs identified extra CNVs, specifically, 10 insertions and 18 deletions within 22q11.2; four were more than 10 kb in size and most contained young AluYs at their breakpoints.To understand the duplication activity leading to the complicated SD structure of this region, we have applied the A-Bruijn graph algorithm to decompose the 22q11.2 SDs to 523 fundamental duplication sequences, termed subunits. Cross-species syntenic analysis of primate genomes demonstrates that many of these LCR22 subunits emerged very recently, especially those implicated in human genomic disorders. Some subunits have expanded more actively than others, and young AluYs are implicated in the past and current duplication events, and moreover suggests that DNA rearrangements in 22q11.2 genomic disorders perhaps do not occur randomly but involve both actively expanded duplication subunits and Alu elements.Our study indicates that Alu repeat elements and AT-rich repeats were enriched and likely involved in many of the past unequal crossover duplications that have shuffled DNAs among blocks and given rise to the current complex genomic architecture of LCR22s .Click here for file"} +{"text": "Appropriate surgical management of colorectal cancers (CRC) in Lynch Syndrome patients, i.e. carriers of germline mutation in a mismatch repair (MMR) gene, is controversial. The decision to remove more or less of the colon requires the consideration of relatively high risk of metachronous CRC with the functional consequence of more extensive surgery. Our aim was to estimate and compare the risks of metachronous CRC for MMR gene mutation carriers following segmental versus extensive removal of colon for their first colon cancer.MLH1, 167 MSH2, 23 MSH6 and 20 PMS2) mutations from the Colon Cancer Family Registry, who had colorectal surgery for their first colon cancer. Age-dependent cumulative risks were calculated using Kaplan-Meier method. Multivariate Cox proportional hazards regression was used to estimate the association between the length of bowel removed and metachronous CRC risk.Risk of metachronous CRC was estimated for 382 carriers of MMR gene (172 P <0.001). Cumulative risk of metachronous CRC was 20% (95%CI 15\u201326%) at 10 years, 44% (95%CI 35\u201355%) at 20 years and 66% (95%CI 52\u201378%) at 30 years after segmental colectomy for a first colon cancer. Risk of metachronous CRC reduced by 24% for every 10 cm of bowel removed. There was no difference in the frequency of colonoscopy after surgery for first colon cancer between extensive and segmental resection (one colonoscopy per 16 (95%CI 13\u201320) months after extensive resection compared to one colonoscopy per 20 (95%CI 18\u201321) months after segmental resection; P 0.2).Of 50 individuals who had extensive colectomy for first colon cancer, none were diagnosed with metachronous CRC over 414 person-years . Of 332 individuals who had segmental colorectal resection for first colon cancer, there were 79 (24%) diagnoses of metachronous CRC over 3,131 person-years . This incidence rate was statistically different from that for individuals who had extensive surgery (Lynch Syndrome patients with colon cancer who have had a segmental colorectal resection have a high cumulative risk for a metachronous CRC. Patients with more extensive resection of the first colon cancer had a lower risk of metachronous CRC compared to less extensive surgery. These findings will better inform decision making about the extent of primary surgical resection."} +{"text": "We evaluated the therapeutic effects of the histone deacetylase inhibitor PXD101 alone and in combination with conventional chemotherapy in treating thyroid cancer.invitro.We studied eight cell lines from four types of thyroid cancer . The cytotoxicity of PXD101 alone and in combination with three conventional chemotherapeutic agents was measured using LDH assay. Western blot assessed expression of acetylation of histone H3, histone H4 and tubulin, proteins associated with apoptosis, RAS/RAF/ERK and PI3K/AKT/mTOR signaling pathways, DNA damage and repair. Apoptosis and intracellular reactive oxygen species (ROS) were measured by flow cytometry. Mice bearing flank anaplastic thyroid cancers (ATC) were daily treated with intraperitoneal injection of PXD101 for 5 days per week. PXD101 effectively inhibited thyroid cancer cell proliferation in a dose-dependent manner. PXD101 induced ROS accumulation and inhibited RAS/RAF/ERK and PI3K/mTOR pathways in sensitive cells. Double-stranded DNA damage and apoptosis were induced by PXD101 in both sensitive and resistant cell lines. PXD101 retarded growth of 8505C ATC xenograft tumors with promising safety. Combination therapy of PXD101with doxorubicin and paclitaxel demonstrated synergistic effects against four ATC lines PXD101 represses thyroid cancer proliferation and has synergistic effects in combination with doxorubicin and paclitaxel in treating ATC. These findings support clinical trials using PXD101 for patients with this dismal disease. The incidence of thyroid cancer has increased over the past three decades worldwide . IncreasHistone deacetylases (HDACs) remove acetyl groups from lysine residues in histone and non-histone substrates, including transcription factors and proteins controlling cell cycle, proliferation and apoptosis. HDACs are divided into 4 classes according to their homology to their yeast orthologues and as appear to be promising targets for cancer therapy -8. Inhib+/I- symporter expression and iodine accumulation could be induced by HDAC inhibitors in poorly differentiated and undifferentiated thyroid cancer . The dose-effect analysis was produced using CompuSyn software , followiFigure S1The combination therapy of PXD101 and chemotherapeutic agents enhances cytotoxicity against ATC. The interactions between PXD101 and chemotherapeutic agents after a 4-day treatment in four ATC cancer lines were evaluated using LDH assays. The combination of PXD101 and chemotherapeutic agents revealed favorable therapeutic effects in all cell lines.(TIF)Click here for additional data file.Figure S2invivo.PXD101 represses RAD51 rapidly and transiently PXD101 greatly reduced RAD51 to less than 21% protein remaining at 3 hours, followed by overexpression of RAD51 from 6 to 24 hours.(TIF)Click here for additional data file."} +{"text": "The aim of this study was to assess the prognostic value of dobutamine stress cardiovascular magnetic resonance (DCMR) in patients with previous myocardial revascularization. The role of DCMR in the risk stratification of patients with previous coronary revascularization has not been well defined.Clinical data and DCMR results were analyzed in 687 consecutive patients with previous percutaneous or surgical coronary revascularisation undergoing DCMR between 2000 and 2004. Follow up was successful for 654 (95.2%) patients. Two hundred and nineteen patients who underwent early revascularisation (\u22643 months) after the test were excluded from analysis. The remaining 435 patients were followed up for a mean of 37 \u00b1 18 months. WMA at rest and the presence of stress-induced WMA (ischemia) were assessed for each patient. Cox proportional hazards regression models were used to identify independent predictors of the composite of cardiac events defined as cardiac death and non-fatal myocardial infarction.Thirty four cardiac events were reported, documented cardiac death in twenty six and non-fatal myocardial infarction in eight patients. In multivariate analysis of clinical data, independent predictors of late cardiac events were the number of dysfunctional segments at rest (hazard ratio (HR) 1.2; 95% confidence interval (CI) 1.1 to 1.3; p<0.001) and stress-induced WMA on DCMR . (Figure Myocardial ischaemia during DCMR is independently predictive of cardiac events among patients with previous myocardial revascularisation."} +{"text": "Etv2, a master regulator of endothelial cell fate, can induce fast skeletal muscle cells to transdifferentiate into endothelial cells in the zebrafish embryo. Etsrp/Etv2 (Etv2) is an evolutionarily conserved master regulator of vascular development in vertebrates. Etv2 deficiency prevents the proper specification of the endothelial cell lineage, while its overexpression causes expansion of the endothelial cell lineage in the early embryo or in embryonic stem cells. We hypothesized that Etv2 alone is capable of transdifferentiating later somatic cells into endothelial cells. Using heat shock inducible Etv2 transgenic zebrafish, we demonstrate that Etv2 expression alone is sufficient to transdifferentiate fast skeletal muscle cells into functional blood vessels. Following heat treatment, fast skeletal muscle cells turn on vascular genes and repress muscle genes. Time-lapse imaging clearly shows that muscle cells turn on vascular gene expression, undergo dramatic morphological changes, and integrate into the existing vascular network. Lineage tracing and immunostaining confirm that fast skeletal muscle cells are the source of these newly generated vessels. Microangiography and observed blood flow demonstrated that this new vasculature is capable of supporting circulation. Using pharmacological, transgenic, and morpholino approaches, we further establish that the canonical Wnt pathway is important for induction of the transdifferentiation process, whereas the VEGF pathway provides a maturation signal for the endothelial fate. Additionally, overexpression of Etv2 in mammalian myoblast cells, but not in other cell types examined, induced expression of vascular genes. We have demonstrated in zebrafish that expression of Etv2 alone is sufficient to transdifferentiate fast skeletal muscle into functional endothelial cells in vivo. Given the evolutionarily conserved function of this transcription factor and the responsiveness of mammalian myoblasts to Etv2, it is likely that mammalian muscle cells will respond similarly. The endothelial cell is a specialized cell type that lines blood vessels. These cells are involved in normal cardiovascular function and become damaged in cardiovascular disease states such as atherosclerosis and stroke. We have discovered that developing muscle cells in the zebrafish embryo can be converted into endothelial cells by the expression of a transcription factor called Etv2. Etv2 normally functions during embryonic development to specify blood and blood vessels. When expressed in muscle cells, Etv2 induces the expression of genes that are normally expressed in endothelial cells; it also represses muscle gene expression. On expressing Etv2, muscle cells change shape and go on to form lumenized blood vessels that connect to the existing circulatory system and support blood flow. The Wnt and VEGF signaling pathways are required for this fate transformation. Our results suggest that muscle cells may be a viable source for the de novo generation of endothelial cells for use in transplantation therapies and they highlight signalling pathways that might be manipulated to improve the efficiency of this process in mammalian cells. In zebrafish, Etv2 is expressed in the lateral plate mesoderm at early somitogenesis stages and defines the first population of angioblasts to arise in the embryo Xenopus causes ectopic expression vascular genes including flk1kdrl, fli1a, and tal1 amongst hundreds of others recently defined by microarray and RNA-seq studies The very early expression of Etv2 and its necessity in vascular development suggest that it may be near the top of the transcriptional hierarchy regulating endothelial cell specification. Overexpression of Etv2 in mouse embryonic stem cells induces the expression of many hematopoietic and vascular genes and increases the yield of these cells when differentiated etv2 and tal1 results in rostral expansion of myocardial precursor cells at the expense of endothelial and primitive myeloid cells etv2 alone is sufficient for expansion of myocardium etv2 and tal1 repress myocardial fates while expanding vascular and primitive myeloid fates Etv2:EYFP transgenic mouse embryos, Rasmussen et al. Tal1 and Fli1 as well as endocardial genes Nfatc1 and Sox17 among the highly expressed genes in this cell population. When Etv2 was knocked out in the EYFP population in vivo, elevated expression of myocardial genes such as Nkx2\u20135, Gata4, and Tbx5 as well as skeletal muscle and smooth muscle genes were noted. In total, all of the studies described above place Etv2 at the top of the genetic hierarchy that defines the balance of endothelial and hematopoietic lineages from cardiac and myogenic lineages in the embryonic mesoderm.A group of recent studies have established that Etv2 is also a critical factor regulating the balance between endocardial versus myocardial cell fates within the anterior lateral plate mesoderm that forms the heart. In zebrafish, knockdown of The regenerative medicine field is very interested in the possibility of generating patient-specific replacement tissues or organs from adult or induced pluripotent stem cells or other cellular sources. Sometimes complex drug and growth factor treatment protocols are necessary to achieve the desired result. However, overexpression of one or a few key developmental transcription factors is often all that is needed to drive a cell from one fate to another. Given Etv2's place atop the genetic hierarchy for the endothelial cell lineage, we were interested in testing the ability of this transcription factor to induce blood vessel formation as a possible treatment for vascular disease.In this study we used zebrafish as a model to demonstrate that overexpression of Etv2 by itself is sufficient to generate functional ectopic blood vessels in vivo. Intriguingly, we found that the additional endothelial cells derived from fast skeletal muscle, revealing an unknown potential for embryonic muscle cells to convert to vasculature. In contrast, two other transcription factors downstream of Etv2 but still near the top of the endothelial lineage, Fli1a and Tal1, are not sufficient to induce blood vessels from muscle. Using time-lapse microscopy of transgenic fish we show that fast muscle fibers change morphology, repress muscle specific genes, and induce vascular genes. Muscle fiber-derived endothelial cells initially change shape, forming large vacuoles and retracting from their long fiber-like shape. They then extend processes and contact other transdifferentiating cells or established endothelial cells, and form patent vessels that support blood flow. To identify signaling pathways supporting this process, we performed a chemical compound screen. We found that precise levels of Wnt signaling are necessary for the efficient induction of vascular genes downstream of Etv2 in transdifferentiating muscle. Additionally, we found that VEGF signaling is necessary for the morphological differentiation of these cells into vessels but is dispensable for the induction of vascular genes by Etv2. Our findings suggest that activation of Wnt signaling coupled with Etv2 expression may be a viable method for inducing transdifferentiation of muscle precursor cells into endothelial cells for therapeutic purposes.hsp70l promoter (named hsp70l:etv2 hereafter). We used the Etv2-mCherry fusion so we could visualize ectopic Etv2 expression directly. We tested the fusion protein for functionality by mRNA injection into kdrl:GFP zebrafish embryos and found that it was as potent as untagged Etv2 at inducing ectopic GFP expression (unpublished data). We confirmed the function of the new transgene by heat shocking double transgenic embryos (hsp70l:etv2 crossed to kdrl:GFP) at dome stage and examining them for ectopic GFP expression. Similar to mRNA-injected embryos, hsp70l:etv2/kdrl:GFP embryos exhibit robust GFP expression restricted mainly to the trunk and tail of embryos that were heat shocked between 24 hpf and 30 hpf . Both transgenic reporters displayed ectopic expression in the trunk following heat shock expression of Etv2, suggesting a broad genetic response was occurring . Surprisingly, kdrl expression was broad at 8 h post\u2013heat shock and not restricted to the trunk as seen with the kdrl:GFP transgene at 24 h. Therefore we performed ISH on 24 h post\u2013heat shock embryos for kdrl and found expression remained fairly broad but was enriched in the trunk . In n\u200a=\u200a20). We also noted that some strongly GFP positive cells located within the fast muscle domain appeared to have reduced or no fast muscle myosin expression by 24 h post\u2013heat shock suggesting these cells might be losing their muscle cell fate through 22 h post\u2013heat shock to red (mCherry+). Hydroxytamoxifen (5 \u00b5M) was added immediately after heat shock to induce Cre activity. Injection of the mylpfa:cre-ERt2 plasmid into ubi:Switch fish resulted in many muscle fibers switching on mCherry expression with little to no nonmuscle switching (unpublished data). In heat-shocked kdrl:GFP/hsp70l:etv2/ubi:Switch embryos, many mCherry+ muscle fibers were observed and 5/15 embryos exhibited +kdrl:GFP, mCherry+ blood vessels (+ blood vessels (0/13) but had robust switching in muscle fibers . By 10 h post\u2013heat shock, +/mylpfa:mRFP+kdrl:GFP cells can be observed although the mylpfa:mRFP is weak due to the rapid decrease in expression following Etv2 expression (+kdrl:GFP cells start taking on a branched morphology and by \u223c40 h these cells take on a lumenized morphlolgy (+mylpfa:mRFP muscle fibers induced kdrl:GFP and 12.5% (11/88 cells) of these cells became part of functional vessels or 3.5% of the total muscle fibers observed (11/312 cells) . Cells t3 cells) . These rkdrl and kdr, VEGF receptors, are induced following Etv2 overexpression in muscle (kdrl:GFP when Etv2 is overexpressed. Inhibition was accomplished in two ways: morpholino-mediated antisense knockdown of the VEGFAa ligand (VEGF-MO) and chemical inhibition of the VEGF receptors with SU5416. Treatment of control embryos with VEGF-MO resulted in lack of intersomitic vessels as reported previously .We also wanted to explore whether increased VEGF signaling could enhance transdifferentiation of muscle into endothelium. To activate VEGF signaling, we chose to overexpress the VEGFAae fibers . VEGF OEat shock and acceat shock . The morFrom these results we conclude that VEGF signaling is not necessary for the induction of transdifferentiation by Etv2 overexpression. However VEGF signaling is necessary for the maturation of these induced cells into endothellial cell-lined vascular networks. Also, activation of VEGF signaling can accelerate the maturation of these induced cells but does not expand the competency for transdifferentiation.kdrl:GFP expression following Etv2 overexpression. We tested multiple pathways including the BMP, Notch, and Wnt pathways described to be upstream of Etv2 in embryonic stem cell cultures We next wanted to know if any common developmental signaling pathways were necessary for Etv2-induced transdifferentiation. Given the zebrafish models utility in drug screens, we decided to test commonly used pathway inhibitors for either increased or decreased hsp70l:etv2/kdrl:GFP fish with XAV939, a chemical inhibitor of the Wnt pathway that acts by indirectly stabilizing the \u03b2-catenin degradation complex by inhibiting tankyrase enzymes that activate the degradation of Axin kdrl:GFP (hsp70l:tcf\u0394C-EGFPhsp70l:tcf\u0394C-EGFP into the hsp70l:etv2/kdrl:GFP fish resulted in a near complete inhibition of ectopic GFP expression following heat shock (kdrl:GFP expression in muscle fibers (hsp70l:ca\u03b2-catenin-2A-TFP) to activate Wnt signaling. Following heat shock, ca\u03b2-catenin suppressed kdrl:GFP expression in muscle fibers nearly as well as Tcf\u0394C and more completely than CHIR99021 . These results demonstrate that a tightly controlled level and timing of Wnt signaling is necessary for fast muscle fibers to transdifferentiate into endothelial cells.Since the response of muscle cells to Etv2 expression changed over time, we reasoned that the cells might also change in responsiveness to Wnt modulation over time. We treated embryos with XAV939 or CHIR99021 and heat shocked them at 22, 24, 26, and 28 hpf. XAV939 significantly inhibited e points . Howevert 28 hpf . This ret 28 hpf . CHIR990t 28 hpf . A dose t 28 hpf . Doses ot 28 hpf . Additiot 28 hpf , suggestIn summary, overexpression of Etv2 can induce fast muscle cells to transdifferentiate into functional endothelial cells in vivo. This competency is developmentally restricted to a time between 22 and 30 hpf and is dependent upon Wnt signaling. Following Wnt-dependent induction, VEGF signaling mediates the maturation and survival of transdifferentiating cells. Following 30 hpf, muscle cells become refractory to Etv2-induced transdifferentiation through an unknown mechanism. Our results are summarized in To determine if mammalian cells have a similar capacity for transdifferentiation, it was tested whether overexpression of Etv2 in the C2C12 mouse myoblast cell line could induce expression of vascular genes. As shown in Here we have described the ability of a single transcription factor, Etv2, to transdifferentiate fast skeletal muscle cells into functional vascular endothelium in vivo. The ability of differentiated muscle cells to become endothelium has not been described before. However, there is a close developmental relationship between these two cell types. In the zebrafish tail, a cell fate choice is made between somite and endothelium and this process is Wnt signaling dependent The developmental window in which we observed this transdifferentiation, 22 hpf to 30 hpf, is quite late in myotome development. Although muscle cells are contractile and expressing muscle-specific genes in this window, they are still undergoing myofusion to become multinucleated and innervation from motor neurons is still expanding within the myotome. Could either of these process be the signal that terminates the competency window for transdifferentiation? This question awaits further study. Interestingly, we observed a wave of competency for transdifferentiation from anterior to posterior. This makes sense as somite development and myogenesis in the trunk normally progress in an anterior to posterior wave. However, wave-like progressions of morphological or signaling events at this late stage of myotome development are not well described. It will be very desirable to identify the factors that define the capacity of fast muscle cells to respond to Etv2.Mesp1 promoter in somites, no fate switch to endothelium occurred Mesp1:Etv2 mice might have been too low to induce fate switching. In either case, if Etv2 is to be used in regenerative medicine applications, its expression will need to be tightly regulated.Etv2 expression is dynamically regulated during development with peak expression when blood and vasculature is being specified Using a targeted small molecule screen, we found that both the VEGF and Wnt signaling pathways were important for muscle cells to become functional endothelial cells. VEGF is a well-established mediator of endothelial cell morphogenesis and survival. We found that VEGF signaling is dispensable for induction of transdifferentiation but mediates the maturation and survival of Etv2-induced muscle cells. Importantly, transcripts encoding the VEGF receptor, Kdrl, did not appear until Etv2-mCherry expression was extinguished in the muscle fibers 8\u201310 h post\u2013heat shock. This most likely corresponds with the onset of VEGF dependency.Canonical Wnt signaling plays important roles in mesoderm specification and the specification of multiple cell types derived from this tissue kdrl:GFP at 8\u201310 h post\u2013heat shock. This defines a small time window from which we can dissect this interaction in the future. Since activation of Wnt signaling alone is not sufficient for transdifferentiation, the Wnt signal must be in parallel or downstream of Etv2. In ES cells overexpression of Etv2 resulted in down-regulation of Wnt-related genes, but no direct interactions were established The point of interaction of Wnt signaling with Etv2 is currently unknown. The function of Wnt signaling in mesoderm development is complicated. Activation or repression of signaling at different developmental stages can result in opposite effects in the same cell population. In mouse ES cells, overexpression of Etv2 can increase the yield of angioblasts and hematopoietic cells A recent report by Ginsberg et al. hsp70l:etv2 transgene, the zebrafish etv2 open reading frame minus the stop codon was cloned into pDONR221 using BP clonase to generate pME-etv2NS. A LR clonase II Plus reaction was then performed with p5E-hsp70l, pME-etv2NS, p3E-mCherrypA, and pDESTTol2pA to generate the Tg(hsp70l:etv2-mCherrypApAT2) plasmid. This plasmid was then injected along with tol2 mRNA (20 pg plasmid and 12 pg mRNA per embryo) into kdrl:GFP+/+ embryos to generate kdrl:GFP/hsp70l:etv2 founder fish. The offspring of injected fish were screened by heat shock for the expression of nuclear mCherry. Three out of 23 screened fish were positive for mCherry expression following heat shock, and of those three, we chose to maintain the line with the strongest expression and the ability to robustly induce kdrl:GFP expression following heat shock at dome stage (see Tg(hsp70l:fli1a-mCherry), Tg, and 121)TgTg transgene was generated using p5E-mylpfa, pME-cre-ERt2, p3E-pA, and pDESTpAT2. Hydroxytamoxifen (5 \u00b5M) was added immediately after heat shock to the fish water to induce recombination in lineage tracing experiments. The Tg(hsp70l:ca\u03b2-catenin-2A-TFP) line was produced using Xenopus laevis \u03b2-catenin with mutations in the GSK3 phosphoryation site that renders it constitutively active hsp70l promoter in a Tol2 vector. Et was isolated by us in an enhancer trap screen (unpublished data). Tg(mylpfa:mRFP) was generated by cloning the mylpfa promoter sequence from genomic DNA into the Tol2mini vector la116Tg(kdrl:GFP)y1Tg(fli1a:EGFP)Tg(ubi:Switch)w74Tg(hsp70l:tcf\u0394C-EGFP)All transgenes were generated using the Tol2kit tage see . The Tg on a Stratagene Mx3005P qPCR system. RNA was isolated using Trizol reagent (Invitrogen) and cDNA generated using Superscript III reverse transcriptase (Invitrogen) with oligo dT primers. Gene expression levels were calculated relative to uninjected controls as previously described kdrl:GFP/hsp70l:etv2/mylpfa:mRFP cells into wild-type embryos was performed as described by Kemp et al. Transplantation of Images were captured on an Axioskop 2 plus microscope (Zeiss) or a Stemi2000-C (Zeiss) using 5\u00d7 or 10\u00d7 objectives with an AxioCam camera and Openlab 4.0 software (Improvision). Confocal images were collected on a Zeiss LSM 510 with a 20\u00d7 water objective. Adobe Photoshop was used to adjust brightness and contrast and assemble composite images.Figure S1Hsp70l:etv2 is as effective as etv2 mRNA injection at inducing kdrl:GFP expression. (A) A shield stage embryo that was heat shocked at dome stage and imaged for mCherry expression. Nuclear-localized Etv2-mCherry is visible. (B) Heat shock of hsp70l:etv2-mCherry transgenic embryos at dome stage induces kdrl:GFP expression at tailbud stage similar to that seen with mRNA injection. (C) mRNA injection of etv2 induces kdrl:GFP expression at tailbud stage. The embryos overexpressing Etv2 in (C) and (D) gastrulated abnormally due to overexpressed Etv2.(TIF)Click here for additional data file.Figure S2qRT-PCR quantification of Etv2-mCherry following heat shock. Etv2-mCherry expression is very high 4 h post\u2013heat shock but quickly decreases to levels similar to control by 24 h post\u2013heat shock. This time course of expression is very similar to that observed by nuclear Etv2-mCherry fluorescence.(TIF)Click here for additional data file.Figure S3Kdrl:GFP, fli1a:EGFP and tal1:GFP are all induced in the trunk of hsp70l:etv2 embryos following heat shock. Hsp70l:etv2 was crossed into kdrl:GFP , fli1a:EGFP , or tal1:GFP , and the resulting embryos were either left at control temperature or were heat shocked at 22 hpf and then imaged at 48 hpf. Control embryos never exhibited ectopic GFP expression in any group. Heat-shocked embryos (+HS) always exhibited ectopic GFP expression in the trunk (right column is high magnification image of the trunk corresponding to the adjacent embryo in the left column), although tal1:GFP was significantly weaker than the other two transgenes. At least 20 embryos were observed for each treatment with similar results.(TIF)Click here for additional data file.Figure S4Fli1a (A), tal1 (B), erg (C), and kdr (D) are all induced in the trunk of embryos 8 h post\u2013heat shock (HS+8 h). Normally these genes are specifically expressed in the vasculature at this time point (control), although tal1 is more strongly expressed in the blood and neurons in the CNS. Note that fli1a and erg have almost ubiquitous expression at this time point, while tal1 and kdr are more restricted to ectopic expression in the trunk. Quantification of the number of embryos demonstrating ectopic expression over the number observed is in the bottom right corner of the corresponding high-magnification trunk image for each group. Eight hours post\u2013heat shock was chosen because it is the time when ectopic expression of kdrl:GFP was first noted in our transgenic analysis.ISH of vascular genes following Etv2 expression. (TIF)Click here for additional data file.Figure S5Myog (A), myf6 (B), mylpfa (C), and tnnt3a (D) are all repressed in the trunk of embryos 4 h post\u2013heat shock (HS+4 h). Normally these genes are strongly and specifically expressed in the musculature at this time point (control). Expression of myog and myf6 is almost completely abolished . Mylpfa and Tnnt3a are reduced but much less so . Quantification of the number of embryos demonstrating normal muscle expression over the number observed is in the bottom right corner of the corresponding high-magnification trunk image for each group. Four hours post\u2013heat shock was chosen since it is the peak of heat shock\u2013induced Etv2 expression.ISH of muscle genes following Etv2 expression. (TIF)Click here for additional data file.Figure S6kdrl:GFP expression at 48 hpf. (A) Heat shock\u2013induced etv2, fli1a, or tal1 are all capable of inducing ectopic +kdrl:GFP cells in the early embryo. Kdrl:GFP embryos were injected with the indicated heat shock\u2013inducible transgenes, heat shocked at shield stage, and imaged at 16 somite stage. Nuclear mCherry (NLS-mCherry) was not able to induce ectopic kdrl:GFP while etv2, fli1a, and tal1 were . The number of embryos exhibiting ectopic GFP expression over the total number observed is represented in the top right corner of each panel. (B) Kdrl:GFP transgenic embryos were injected with hsp70l:etv2, hsp70:fli1a, or hsp70:tal1 transgenes and heat shocked at 24 hpf. Each transcription factor was labeled with mCherry and expression was confirmed by imaging 3 h post\u2013heat shock (inset). GFP expression was imaged at 48 hpf. Etv2 overexpression resulted in strong ectopic GFP expression, but neither Fli1a nor Tal1 was sufficient for inducing the same response. The ratio in the bottom right corner of each panel represents the GFP positive embryos over the total observed.Fli1a and Tal1 overexpression at 24 hpf is not sufficient to induce ectopic (TIF)Click here for additional data file.Figure S7Slow muscle fibers do not respond to Etv2 overexpression. Immunostained sections through the trunk of 48 hpf hsp70l:etv2/fli1a:EGFP embryos that were untreated (control) or heat shocked at 24 hpf (HS+24 h). Sections were stained for GFP and slow muscle myosin. Nuclei are stained with DAPI in the mergeDAPI panels. fli1a:EGFP is normally expressed in the intersomitic vessels (ISVs) and axial vessels (AVs) of control sections. No co-staining of GFP and slow muscle myosin was observed (arrows). ROI is the region of interest highlighted by the dashed box in each panel. One section from 20 different embryos was observed for each treatment group with similar results within each group.(TIF)Click here for additional data file.Figure S8mylpfa:mRFP is co-expressed with kdrl:GFP following overexpression of Etv2. (A) Confocal image of the trunk of a kdrl:GFP/mylpfa:mRFP/hsp70l:etv2 triple transgenic embryo heat shocked at 24 hpf and imaged at 12 h post\u2013heat shock. A GFP/mRFP double positive muscle fiber is highlighted by the arrow and x-axis and y-axis z-plane projections are presented below and to the right of the image respectively. (B) Fast muscle myosin and GFP colocalize in the trunk of kdrl:GFP/hsp70l:etv2 transgenic fish heat shocked at 24 hpf and imaged at 48 hpf. Red muscle fibers colocalize with GFP (white arrows). A strongly GFP positive cell located where a muscle fiber normally would be is negative for fast muscle myosin (large white arrowhead), suggesting this cell has lost its muscle cell identity.Fast muscle\u2013specific (TIF)Click here for additional data file.Figure S9kdrl:GFP expression. (A) Time lapse imaging of an intersegmental vessel angiogenic sprout from hsp70l:etv2 embryos prevent ectopic kdrl:GFP expression. Embryos were initially heat shocked at 24 hpf and then either maintained at normal temperatures or were treated with heat shock every 12 h the indicated number of times. Embryos treated with multiple heat shocks displayed abnormal morphology whose severity correlated with the number of heat shocks, including cardiac edema suggestive of circulatory failure (left column and inset). Control embryos heat shocked four times appeared similar to non\u2013heat shocked embryos, HS(\u2013). When Etv2 expression was maintained for the whole period (4\u00d7 heat shocks), kdrl:GFP was not ectopically induced and was reduced or absent in the normal vasculature (right column).Etv2 overexpression is toxic to angiogenic sprouts and its maintained expression prevents (TIF)Click here for additional data file.Figure S10mylpfa:cre-ERt2 injected kdrl:GFP/hsp70l:etv2/ubi:Switch fish demonstrates endothelial cells derived from muscle fibers (arrow). The ubi:Switch transgene changes from GFP to mCherry following recombination initiated by Cre. The mylpfa promoter specifically drives Cre expression in fast muscle fibers. In non\u2013heat shocked embryos (\u2212HS) only mCherry muscle fibers are observed (n\u200a=\u200a13), while following heat shock mCherry+, kdrl:GFP+ vessels were observed in 5 out of 15 embryos observed. Note that the kdrl:GFP transgene in the vasculature is significantly brighter than the ubi:Switch GFP+ background. All embryos were treated with hydroxytamoxifen (5 \u00b5M) immediately after heat shock or the equivalent time for non\u2013heat shocked controls.Lineage tracing of muscle cells demonstrate they are a source of new blood vessels following Etv2 expression. Lineage tracing of fast muscle fibers in (TIF)Click here for additional data file.Figure S11mylpfa:mRFP/hsp70l:etv2/kdrl:GFP+, into wild-type embryos. Approximately 10 cells were transplanted per embryo. Transplanted embryos were raised until 22 hpf at which point they were selected for embryos displaying mylpfa:mRFP expression in distinct regions absent in kdrl:GFP. These embryos were then either heat shocked or left as no heat shock controls. Embryos were then analyzed for mylpfa:mRFP/kdrl:GFP coexpression at 10 h post\u2013heat shock and followed out to 44 h post\u2013heat shock. Two kdrl:GFP positive muscle fibers, one still mylpfa:mRFP positive (arrow), under-go transdifferentiation to form functional vessels supporting blood cell flow (see Movie S4).Etv2 cell autonomously initiates transdifferentiation of muscle cells. Blastula cell transplantation was performed from triple transgenic, (TIF)Click here for additional data file.Figure S12Hsp70l:etv2/kdrl:GFP embryos were heat shocked at 28 hpf, treated with various doses of CHIR99021, and GFP+ myotomes were quantified at 48 hpf. Doses between 5 and 25 \u00b5M significantly increased the number of GFP+ myotomes but not to the levels seen when heat shock was administered at earlier time points. t test, (*) p<0.05, (**) p<0.01.Activation of Wnt signaling with different doses of CHIR99021 can expand transdifferentiation at 28 hpf. (TIF)Click here for additional data file.Figure S13Etv2 induces vascular marker gene expression in C2C12 cells but not in other cell types. (A) Induction of endothelial cells marker expression in myoblast cell line C2C12. Fli1, Scl, Kdr, and vascular endothelial cadherin (Vecad) expression levels detected by qPCR were compared between Etsrp/Etv2 transfection and control mCherry transfection. Fold of induction compared to nontransfected cells is shown. (B) PCR detection of vascular marker expression in three tumor cell lines that were transfected with Etv2. No induction of the listed endothelial cell markers was detected.(TIF)Click here for additional data file.Movie S1Kdrl:gfp/mylpfa:mRFP/hsp70l:etv2 non\u2013heat shocked control.(MOV)Click here for additional data file.Movie S2Kdrl:gfp/mylpfa:mRFP/hsp70l:etv2 heat shocked at 24 hpf and imaged from +8 h to +22 h.(MOV)Click here for additional data file.Movie S3Timelapse imaging of a single fast muscle fiber becoming kdrl:GFP positive and changing morphology. This movie corresponds to the right panels of (MOV)Click here for additional data file.Movie S4+kdrl:GFP cells in the upper left are muscle derived and support blood circulation.Blood flow through muscle-derived endothelial cells in a chimeric embryo. Video corresponds to the region highlighted in (MOV)Click here for additional data file.Table S1Gene-specific primers for qPCR.(DOCX)Click here for additional data file."} +{"text": "The presens of myocardial scar tissue as a potential arrhythmogen substrate may influence the treatment of patients with coronary artery disease (CAD). Delayed enhancement MRI (DE-MRI) allows accurate detection and visualization of myocardial scar tissue. We thought to detect myocardial scar tissue in patients with CAD and preserved left ventricular (LV) function without regional wall motion abnormalities (RWMA) in transthoracic echocardiography (TTE) using DE-MRI.83 patients with a history of CAD and with normal LV function without RWMA in TTE were examined for viability determination using DE-MRI .45 54.2%) of these patients suffered from an acute coronary syndrome shortly before the MRI examination while 8 patients were found with a history of ACS further in the past. Myocardial scar tissue was found in 57 patients (68.7%), whereas 26 patients (31.3%, Figure % of thesFrom our preliminary results, a significant numbers of patients with CAD, preserved LV function and without regional wall motion abnormalities suffer from myocardial scar tissue. DE-MRI examinations of the heart for determination of myocardial viability should be considered in these patients to assess the extent of myocardial injury as TTE might underestimate the extent of myocardial scar tissue in patients with preserved left ventricular function.none"} +{"text": "The device provides proof of concept for programmable, function-independent DNA selection in vivo and provides a unique example of a logical-functional interface of an engineered synthetic component with a complex endogenous cellular system. Further research into the design, construction and operation of synthetic devices in vivo may lead to other functional devices that interface with other complex cellular processes for both research and applied purposes.The extraordinary fidelity, sensory and regulatory capacity of natural intracellular machinery is generally confined to their endogenous environment. Nevertheless, synthetic bio-molecular components have been engineered to interface with the cellular transcription, splicing and translation machinery in vivo by embedding functional features such as promoters, introns and ribosome binding sites, respectively, into their design. Tapping and directing the power of intracellular molecular processing towards synthetic bio-molecular inputs is potentially a powerful approach, albeit limited by our ability to streamline the interface of synthetic components with the intracellular machinery in vivo. Here we show how a library of synthetic DNA devices, each bearing an input DNA sequence and a logical selection module, can be designed to direct its own probing and processing by interfacing with the bacterial DNA mismatch repair (MMR) system Although the cellular machinery is orders of magnitude more complex than any synthetic biological device produced so far Synthetic biology often assimilates existing knowledge gained through basic research into new functional synthetic components and/or systems We introduce an approach to engineer a synthetic DNA device which directs the probing and processing power of the endogenous MMR machinery towards the selection of a specific, but arbitrary DNA sequence according to rules embedded within the device's sequence and structure . More coThe device is a circular dsDNA molecule with twoThe Selection module is embedded within the coding region of an Ampicillin (Amp) resistance gene and contains two functional elements : (1) a lMore concretely, upon transformation of the device to bacteria the MutHLS machinery detects whether a mismatch is present in the input module. Positive (mutation) diagnosis induces (1) MMR scanning for the closest hemi-methylated Dam site within a 1 Kb range Description of experimental procedures .Error-prone PCR was done using GeneMorph II Random Mutagenesis Kit (Stratagene) according to standard protocol except for Thermal Cycler program: Activation 95\u00b0C for 6 min, 15\u201319 cycles of: Denaturation 95\u00b0C for 30 sec, annealing at 55\u00b0C for 30 sec and extension 72\u00b0C for 1\u223630 min.On ice, 1\u20132 \u00b5l of purified DNA, eluted in double deionized water (DDW) was mixed with 25 \u00b5l of electrocompetent bacterial cells. The mixture was transferred into an electroporation cuvette (BTX) followed by the employment of a 1.8 kV pulse using the Gene Pulser Xcell total system (Bio-Rad). After electroporation, 975 \u00b5l of SOC were added and a recovery step of 1 hour at 37\u00b0C was performed before inoculation into selective petri dishes.We used Sanger sequencing using the BigDye\u00ae Terminator v1.1 Cycle Sequencing Kit (ABI). We purified the sequencing reaction using the Performa\u00ae DTR Ultra 96-Well Plate Kit (EdgeBio) and analyzed the products using the ABI 3130 genetic Analyzer.See Mismatches between the various DNA molecules cloned into the input module were exposed by melting and re-annealing the input modules, which generated both hetero and homo-duplex input modules. The fractions of homo-duplex input modules are known since the concentrations of each input module variant in the mixture were controlled by us. We calculate that the small differences in sequence between input modules had a negligible effect on the Tm between the different variants (see sequences in de novo DNA construction (i.e. when there are many more erroneous than error-free molecules) in vivo or in culture in vivo operation of the device is re-annealing dependent and, at the same time, integrated a functional Kanamycin resistance gene into a different location on the device. This eliminated false positive colonies originating from inefficient Selection module integration and/or its incomplete restriction out of the device prior to its integration, while enabling propagation the device for preparative purposes using Kan selection. We added a control step for the proper digestion of the vector by the restriction enzymes , optimizin vivo using a library of 38 erroneous GFP gene variants generated randomly by error-prone PCR and an additional variant for which we attempted to enrich to the molecules in the library, thereby enriching its fraction within the homoduplex population.Figure S1In silico simulation of enrichment potential. The initial fraction of devices with an error free input module (X axis) is plotted against the fraction of devices with an error free, homo-duplex input module out of the total population of homo-duplex devices. The curves (from right to left) represent increasing numbers of initial devices with erroneous DNA inputs. These graphs exemplify the fact that the enrichment factor of our system increases with library size.(TIF)Click here for additional data file.Figure S2Comparison between the enrichment factor of W.T and Dam- strains. Comparison between the enrichment factor of from two bacterial strains, E.cloni (Lucigen) and GM48, with and without a functional MMR system (Dam-), respectively. (A) Comparison between E. cloni enrichment factor negative control and operative device (colonies tested: n\u200a=\u200a47 and n\u200a=\u200a47 respectively). (B) GM48 enrichment factor comparison between negative control and a functional device (colonies tested: n\u200a=\u200a92 and n\u200a=\u200a80 respectively).(TIF)Click here for additional data file.Figure S3Capillary electrophoresis analysis of digestion with MboI. MboI is a restriction enzyme which digests dam sites (GATC) but is blocked by methylated and hemimethylated dam sites. We used MboI assay to test the methylation efficiency of in the Selection site (A) Digestion of dsDNA constructed of (1) the specially modified methylated strand, and (2) a complementary unmethylated oligonucleotide, labeled by HEX fluorophore. (TIF)Click here for additional data file.Figure S4pIVEC (\u223c4000 bp) restriction enzymes control. We have digested the vector with different restriction enzymes to verify their proper activity. Lane 1: undigested plasmid (supercoiled) lane 2: XhoI & SspdI (KasI), expected size \u223c1050; lane 3: XhoI & HindIII, expected size \u223c1000; lanes 4, 5 and 6: Single digestion using XhoI, SspdI (KasI) & HindIII respectively; lane 7: Digestion using SspdI (KasI) & HindIII , expected size \u223c50 bp.(TIF)Click here for additional data file.Figure S5Colony count results. (A) Series name indicates Selection site: Vector ratio in ligation reaction and x-axis determine the type of Selection site structure Tra(TIF)Click here for additional data file.Figure S6Example of sequencing analysis of a colony containing two different DNA molecules. Two kinds of populations are detectable by sequencing the plasmid from one direction: A division to two kinds of sequences is exhibited at the start location of the loop structure (heteroduplex DNA)..(TIF)Click here for additional data file.Table S1Mutation analysis of GFP variants, produced using error-prone PCR, compared with error-free GFP sequence reference.(PDF)Click here for additional data file.Table S2Selection sites structures which serve as controls for experiments and their expected result after transformation to E. coli. (A) positive control - restores the ampicillin resistance, bacteria should live; (B) negative control - inserts a frame shift to \u03b2-lactamase gene, bacteria should die; (C) the tested system with a functional selection site, bacteria should live only if no mismatch was found (See ound See .(TIF)Click here for additional data file.Table S3Selection sites structures and their expected viability decision, in transformed E. coli.(TIF)Click here for additional data file.Text S1Supplementary methods.(DOC)Click here for additional data file.Text S2Supplementary sequences.(DOC)Click here for additional data file.Text S3In silico simulation of enrichment potential.(DOC)Click here for additional data file.Text S4Device operation in wild type Vs. Dam deficient bacteria - In vivo Dam methylation may reprogram the device.(DOC)Click here for additional data file.Text S5Methylation evaluation experiment - Selection module hemi-methylation is highly efficient.(DOC)Click here for additional data file.Text S6Restriction control for the insertion of the Selection module.(DOC)Click here for additional data file.Text S7Selection module loop optimization - Integration of the Selection module loop into the device can be optimized.(DOC)Click here for additional data file.Text S8Invisibility of the selection module loop to the MMR system -The selection module loop is partially invisible to the MMR system.(DOC)Click here for additional data file.Text S9Generating the library of input modules using error-prone PCR.(DOC)Click here for additional data file.Text S10Description of experimental procedures.(DOC)Click here for additional data file."} +{"text": "Macrophages play a critical role in innate immunity, and the expression of early response genes orchestrate much of the initial response of the immune system. Macrophages undergo extensive transcriptional reprogramming in response to inflammatory stimuli such as Lipopolysaccharide (LPS).TNF), was characterized by H3Ac, high CpG content and lack of TATA boxes. The second group, containing inflammatory mediators (interleukins and CCL chemokines), was up-regulated upon LPS stimulation despite lacking H3Ac in their annotated promoters, which were low in CpG content but did contain TATA boxes. Genome-wide analysis showed that few H3Ac peaks were unique to either +/\u2212LPS condition. However, within these, an unpacking/expansion of already existing H3Ac peaks was observed upon LPS stimulation. In contrast, a significant proportion of S5P RNAPII peaks (approx 40%) was unique to either condition. Furthermore, data indicated a large portion of previously unannotated TSSs, particularly in LPS-stimulated macrophages, where only 28% of unique S5P RNAPII peaks overlap annotated promoters. The regulation of the inflammatory response appears to occur in a very specific manner at the chromatin level for specific genes and this study highlights the level of fine-tuning that occurs in the immune response.To identify gene transcription regulation patterns involved in early innate immune responses, we used two genome-wide approaches - gene expression profiling and chromatin immunoprecipitation-sequencing (ChIP-seq) analysis. We examined the effect of 2 hrs LPS stimulation on early gene expression and its relation to chromatin remodeling and promoter binding of Sp1 and RNA polymerase II phosphorylated at serine 5 (S5P RNAPII), which is a marker for transcriptional initiation. Our results indicate novel and alternative gene regulatory mechanisms for certain proinflammatory genes. We identified two groups of up-regulated inflammatory genes with respect to chromatin modification and promoter features. One group, including highly up-regulated genes such as tumor necrosis factor ( Macrophages play a critical role in innate immunity, and recognize \u2018microbial non-self\u2019 via receptors known as toll-like receptors (TLRs) that recognize pathogen-associated molecular patterns (PAMPs) The expression and repression of genes is associated with alterations in chromatin structure mediated by enzymatic modifications of the N-terminal ends of the core histones in nucleosomes The recruitment of the RNA polymerase II (RNAPII) in the core promoter is essential for gene expression. However, there is not necessarily a correlation between the rate of its recruitment and mRNA levels of a gene Studies of gene regulation have advanced rapidly in recent years, and this is due to the development of next generation sequencing techniques. Chromatin immunoprecipitation followed by massively parallel sequencing (ChIP-seq) has become a valuable experimental tool for examining protein-DNA interactions 2. For differentiation of THP-1 cells into macrophages a protocol using conditioned media as previously described by Whatling et al CD11b expression was measured using FACS (THP-1 cells (monocyte cell line) were maintained in culture in RPMI 1640 medium containing 10% fetal bovine serum, 1 mM sodium pyruvate, 100 units/ml penicillin and 100 \u00b5g/ml streptomycin at 37\u00b0C with 5% COing FACS and quaning FACS .Escherichia coli O55:B5 for 2 hours to induce an acute inflammatory response. To confirm an induction of the inflammatory response in the THP-1 macrophages, TNF mRNA levels were measured in +/\u2212LPS stimulated samples (THP-1 macrophages were then either left unstimulated (\u2212LPS) or stimulated with 1 \u00b5g/ml LPS (+LPS) from samples .6) were incubated for 2 hours and total mRNA was extracted using TRIZOL\u00ae reagent and purified using the RNeasy Mini Kit following the manufacturer's instructions, including DNase treatment . RNA concentration and quality were analyzed using Nanodrop 1000 Spectrophotometer and Agilent 2100 Bioanalyzer respectively. The RNA was then hybridized and scanned on the Affymetrix\u00ae Genechip\u00ae Human Exon 1.0 ST array following manufacturer's protocols at the Karolinska Institute microarray core facility. Standard Affymetrix quality controls were followed (http://www.genomatix.de).THP-1 macrophages (0.5\u00d710followed . The subChromatin immunoprecipitation (ChIP) was performed on differentiated THP1 macrophage cells +/\u2212LPS stimulation for 2 hours using a method previously described by Rada-Iglesias et al http://main.g2.bx.psu.edu/) Sequencing was performed on the Illumina Genome Analyzer I . The data is available from gene expression omnibus under accession number GSE32325. All reads that passed mapping quality checks were anain silico analysis and a matrix similarity value of 0.9 was used as the cut-off point.TATA-box patterns were identified in the promoter sequences of genes downloaded from the UCSC browser. Using GENOMATIX, classical TATA boxes matrices were used for Using the genes identified in the microarray gene expression analysis, a list was generated of those that had peaks that were replicated in the biological replicas. Data for CpG island coordinates were downloaded from the UCSC genome browser CD11b expression using FACS TSS position, annotated transcription start site (hg19); H3AC common peak: presence (YES) or absence (NO) of a significant H3Ac peaks in promoter (\u22122 Kb/+1 Kb of TSS) for both +/\u2212LPS conditions.; the effect of LPS, Peak extension; direction of chromatin expansion ; S5P RNAPII & Sp1 peaks: presence of a significant peak in unstimulated condition (\u2212LPS), LPS treated cells (+LPS); common in both conditions (+/\u2212LPS) or absence in either condition (NO) in promoters (\u22122 Kb/+1 Kb of TSS); CpG island presence or absence in promoter (\u22122 Kb/+1 Kb of TSS); TATA box in silico finding (\u2212150 bp/+50 bp of TSS). RED BOLD: 105 genes included in MeSH inflammatory category. Tables are divided into gene families and classified groups.(DOC)Click here for additional data file.Table S7MatchInspector in silico TATA Box finding. Tables show the results of in silico TATA box finding in the subset of up- and down-regulated genes upon LPS stimulation in a distance of \u2212150 bp/+50 bp of RefSeq gene annotated TSS. Gene symbol, Position of the TATA box from 5\u2032UTR in base pairs (bp), gene strand direction , promoter sequence (Lowercase DNA sequence letters: promoter area (Upstream 5\u2032UTR), Uppercase DNA sequence: 5\u2032UTR; and Underlined and in bold: TATA box sequence), TATA box category , and matrix similarity (cut off \u22650.9). .(DOCX)Click here for additional data file."} +{"text": "Effect of long term cholesterol diet withdrawal on accelerated atherosclerosis in iliac artery of New Zealand White (NZW) rabbits has not been explored so far. Atherosclerosis was thus induced in rabbits by a combination of balloon injury and atherogenic diet (AD) (1% cholesterol and 6% peanut oil) feeding for 8 weeks (baseline) followed by chow diet (CD) feeding for 4, 8, 16, 32, 50 and 64 weeks. The plaque characterization was done using histology, real time RT-PCR and vasoreactivity studies. Significant elevation in plasma lipids with AD feeding was normalized following 16 weeks of CD feeding. However, baseline comparison showed advanced plaque features even after 8 weeks of CD period with significant elevation in intima/media thickness ratio and plaque area later showing reduction at 50 and 64 weeks CD periods. Lesion lipid accumulation and CD68 positivity was maintained till 16 weeks of CD feeding which significantly reduced from 32 to 64 weeks CD periods. Baseline comparison showed significant increase in ground substance, MMP-9 and significant decrease in \u03b1-actin and collagen content at 8 weeks CD period indicating features of unstable plaque. These features regressed up to 64 weeks of CD. Partial restoration of functional vasoconstriction and vasorelaxation was seen after 64 weeks of CD feeding. mRNA expression of MCP-1, VCAM-1, collagen type I and III, MMP-9, TIMP-1, IFN-\u03b3, TNF-\u03b1, IL-10 and eNOS supported the above findings. The study thus reveals insights into initial plaque instability and subsequent regression on AD withdrawal in this model. These results are suggestive of an appropriate window for drug intervention for plaque stability/regression and restenosis as well as improves understanding of plaque regression phenomenon in this model. Atherosclerosis is the predominant underlying pathology of cardiovascular disease, the most common cause of premature death in the industrialized world Animal experimental protocols were approved by the Institutional Animal Ethical Committee, Council for Scientific and Industrial Research- Central Drug Research Institute (CSIR-CDRI) . Procedures were carried out in strict accordance with the Guidelines of Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) which conforms to the international norms of Indian National Science Academy (INSA). All efforts were made to improve animal welfare and minimize suffering.Male New Zealand White rabbits with an initial body weight of 2.0\u20132.5 kg, maintained at Central Animal House facility of CSIR-CDRI, Lucknow were used. All animals consumed an atherogenic diet (1% cholesterol and 6% peanut oil) for 8 weeks to induce atheroma formation ad libitum chow diet (CD) and atherogenic diet (AD) for 8 weeks respectively. Animals of the normal group (n\u200a=\u200a7) received CD for 8 weeks and were immediately sacrificed thereafter. At the end of 8 weeks period of AD feeding, sixty four animals were divided into seven groups on the basis of similar ranges of hypercholesterolemia. Baseline group animals were sacrificed immediately after the 8 weeks of AD feeding, while other groups were sacrificed after AD cessation and CD resumption for 4 weeks , 8 weeks , 16 weeks , 32 weeks , 50 weeks and 64 weeks respectively , triglycerides (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) were estimated using Automated analyzer using commercial kits At termination, iliac artery portion near to aorta-iliac bifurcation was isolated and preserved in 10% buffered formalin solution for 24 hours. One proximal portion of the artery was frozen in OCT compound for cryostat sectioning and other portion was processed for paraffin embedding. Arteries of all groups underwent simultaneous and similar fixing, dehydration and wax infiltration procedures to nullify the dimension changes induced by solvent treatments. Serial paraffin sections (5 \u00b5m) were collected throughout arterial length for staining procedures. For general architecture of plaque, Hematoxylin and Eosin (HE) staining and for cellular components Movat Pentachrome staining wherein muscle is stained as red, fibrinoid tissueshows as intense red, mucin/ground substance is stained blue to blue green, black color reflects nuclei and elastic fibres and yellow color depicts collagen and reticular fibres2), plaque area (PA) (mm2) and percentage cross sectional narrowing (%CSN) was carried out in at least 25\u201330 sections throughout the arterial segment by blinded investigators employing computer-assisted image analysis software on Movat Pentachrome-stained arterial sections . Intra-observer variation was less than 5%. The intimal thickness was measured by drawing lines perpendicular to the lumen at five different locations of a stained section and the mean value was calculated Structural changes in lesions of each group of animals were assessed in consecutive sections stained with Movat Pentachrome. The following features were employed for describing general lesion morphology of respective groups: i) internal elastic lamina (IEL) rupture , ii) media atrophy, iii) media breakdown, iv) adventitial breakdown and v) inflammation Paraffin-embedded consecutive sections were stained with the following antibodies: Mouse monoclonal anti rabbit CD68 , alpha smooth muscle actin and MMP-9 . Immunoreactivity was revealed using the Novacastra Novolink Polymer detection system and counterstained with hematoxylin. Appropriate positive control was used for all antibodies. Negative controls included omission of the primary antibody and use of non-immune mouse IgG or secondary antibody only. In all cases, negative controls showed insignificant staining.2\u20135% CO2. The tissue was kept under a constant tension of 2 g throughout the experiment and was equilibrated by intermittent changing of Kreb's buffer for 90 min 4 mm long iliac artery segments were mounted in a 10 ml organ bath containing Krebs solution, maintained at 37\u00b0C and continuously oxygenated with 95% O\u0394\u0394Ct) method using GAPDH as an internal standard Quantitative gene expression analysis was performed by using SYBR Green technology. Total RNA was extracted from freshly isolated injured iliac artery of different groups using TRIZOL isolation procedure as described previously The results were calculated as mean \u00b1 SEM. The statistical significance of difference between different groups was determined by one way ANOVA followed by Dunett's post-hoc test using GraphPad Prism 5 software. P value of 0.05 or less was chosen for statistical significance.vs baseline) and Reg 8 week groups respectively, although it was still elevated in comparison to normal group. Aged animals till 5 years of age fed chow diet do not exhibit any significant changes in plasma lipid levels in comparison to young rabbits The baseline group showed significant increase in TC, TG, LDL-C and HDL-C (p<0.001) compared to normal animals. During regression period, plasma cholesterol levels dropped significantly in Reg 4 week and CD68 positive cells was observed both in the neointimal area towards lumen as well as towards IEL. Interestingly, compared to baseline group, IMT ratio and plaque area was significantly increased (p<0.001) at Reg 8 week group signifying enhanced plaque burden even after 8 weeks of AD withdrawal. The percentage lipid area and CD68 positive area was also maintained up to the baseline group levels in this group, which was progressively decreased in late regression groups (Reg 32 week to Reg 64 week) (vs baseline at Reg 64 week) (vs baseline) respectively along with corresponding decrease in percentage cross sectional narrowing (p<0.01 vs baseline) with no incidence of foam cell formation. In baseline group, atherosclerotic lesion showed significant increase in arterial IMT ratio, plaque area and % CSN (p<0.001) and significant decrease in lumen area (p<0.001) compared to normal animals . The bas64 week) . The res64 week) . Concomiaseline) . These rvs normal) thus depicting enhanced ground substance accumulation was increased in baseline group (vs baseline) and MMP-9 protein (p<0.01 vs baseline) and its mRNA expression pattern demonstrating a tendency for plaque stabilization and its evolution was also ascertained using Movat Pentachrome and MMP-9 staining in serial sections in respective groups. Ground substance is composed of extracellular matrix components including proteoglycans and other matrix substances including MMPs in a proenzyme form responsible for lipid entrapment and plaque instability mulation . MMP-9 mne group . Comparene group with sublization . The resvs normal) hence depicting a fibromuscular hyperplasia response (vs baseline) (Plaque collagen and smooth muscle content are known to provide stability to atheromatous lesions and prevent their rupture response . Concomiaseline) and collaseline) . CumulatThe atherosclerotic plaque in baseline group was presented as fibrocellular hyperplasia. In baseline group, the lesion prominently composed of foam cells occupying on an average 10% to 15% of the plaque area, with the remainder of the plaque area being occupied by smooth muscle cells and fibrous tissue matrix. There was general incidence of IEL duplication at the site of IEL breakage and medivs normal) (vs baseline) along with significant decrease in eNOS mRNA expression (vs baseline), signifying revival of functional endothelium . The vasaseline) . Likewis normal) . Howeverothelium . This efothelium .To explore possible cause or effect of plaque regression on key genes involved in atherosclerosis, we did real time mRNA expression of atherosclerotic iliac arteries. The baseline group showed enhanced mRNA expression of VCAM-1 (4 fold), MCP-1 (4 fold), TNF-\u03b1 (4 fold), TGF-\u03b21 (10 fold) in comparison to normal . Howevertunica intima due to a complex array of molecular and cellular events. However, depending on their surrounding milieu, atherosclerotic plaques might progress, stabilize or regress Atherosclerosis is a focal pathology that occurs as an asymmetric thickening of the arterial milieu as shown by increased mRNA expression of TNF-\u03b1, IFN-\u03b3 and MCP-1. A corresponding decrease in alpha smooth muscle actin and collagen content was also observed in this group. Hence, the plaques developed in this group fulfilled some criteria of an unstable human atherosclerotic plaque Primarily, microscopic level regression in atherosclerosis includes (1) arrest of intimal cell proliferation; (2) restored integrity of the endothelium lining the plaques; and (3) decrease in the amount of intracellular and interstitial lipid A prominent similarity between rabbit and human lesions is the presence of foam cells Collagen and muscle content in plaques and their reorganization is suggested to play a pivotal role in maintaining plaque stability via SMC migration viz. Reg 4 week and Reg 8 week. The results are in tune with previously published observations in rabbits milieuBlood flow variations may influence the relative residence time of lipoproteins, blood borne molecules and inflammatory cells in various arteries thus altering gene expression profile via reduced eNOS expression via various mechanisms. Results obtained indicate recovery of functional responses of endothelium during plaque regression in Reg 50 week and Reg 64 week groups which might be due to availability of NO via increased eNOS expression. This needs to be confirmed in future studies. It is plausible that the migration of arterial ECs from adjacent intact endothelium (from aorta near iliac bifurcation) or bone marrow-derived endothelial precursor cells might be involved in endothelial regeneration Endothelial regeneration is one of the necessitating factors for atherosclerosis regression. In our experimental model, balloon injury is the key triggering and pronounced feature for removal of endothelium. Endothelial cell (EC) injury in this setting is more pronounced and needs a larger area to be regenerated especially in the setting of hyperlipidemia as in our case. The impairment in endothelium dependent relaxation in baseline group may be due to reduced elaboration or increased degradation of endothelium derived NO via alteration of specific pro-atherogenic cellular and molecular pathways. The results support current clinical concepts that long-term plaque reorganization leads to plaque stabilization due to normalized atherogenic conditions. These components appear to be completely reversed in humans who have substantial and sustained lowering of their plasma lipids One needs to appreciate that plaque regression is not simply a rewinding of the sequences of lesion progression, but instead involves mobilization of plaque pathological components Currently, there is large unmet need for a direct disease modifying drug in atherosclerosis. One of the reasons might be due to lack of complete pathological understanding of atherosclerosis at various artery sites in preclinical models thus resulting in clinical failure of investigative agents. Hence, there is an urgent need to characterize atherosclerotic features at various artery sites in animal models and interpret results from that perspective. Majority of arteries employed for atherosclerosis studies in rabbits are carotid, aorta, iliac and femoral. A head to head comparison of atherosclerosis presentation at these sites under similar experimental conditions will not only help to maintain uniformity in preclinical understanding of artery specific changes but also accelerate atherosclerosis drug discovery.The study results indicate that atherosclerotic iliac artery of NZW rabbits presents features of human unstable plaques after a brief period of AD removal. Also, collective changes in plaque intracellular and extracellular lipid, restoration of endothelium functionality, increase in fibrous component (collagen and SMCs), decreased metalloproteinase and modulation of gene expression orchestrated plaque regression associated with this model . The stuFigure S1Histological examples of morphological changes in rabbit plaques of various groups stained with Movat Pentachrome. A\u200a=\u200aIEL duplication at the site of IEL breakage , B\u200a=\u200a Medial elastic fibres duplication , C\u200a=\u200a Focal medial breakdown with IEL breakdown (Baseline group section), D\u200a=\u200aMedial fibrotic reaction (Baseline group section), E\u200a=\u200a Lipid core with extracellular lipid and foam cell deposition (Baseline group section), F\u200a=\u200a Medial breakdown with no EEL breakage (Reg 8 week section), G\u200a=\u200aFocal medial compression extending towards atrophy (Reg 8 week section), H\u200a=\u200aMedial atrophy (Reg 8 week section), I\u200a=\u200aIEL breakage with no damage to media (Reg 64 week section) and J\u200a=\u200a Focal cholesterol clefts (Reg 50 week and Reg 64 week sections). Scale bar\u200a=\u200a50 \u00b5m.(TIF)Click here for additional data file.Figure S2mRNA expression of key genes involved in atherosclerosis as determined by real time PCR. (A) VCAM-1 (B) MCP-1 (C) TNF-\u03b1 (D) IFN-\u03b3 (E) TGF-\u03b21 (F) IL-10. *p<0.05 and ***p<0.001 vs normal; #p<0.05, ##p<0.01 and ###p<0.001 vs Baseline.(TIF)Click here for additional data file.Table S1Oligonucleotide primers and specific annealing temperatures for real-time RT-PCR.(DOC)Click here for additional data file."} +{"text": "One specific ZFN pair facilitated non-homologous end joining (NHEJ)-mediated gene disruption when expressed in mouse zygotes. We also observed a single homologous recombination (HR)-driven gene replacement event when this ZFN pair was co-injected with a targeting vector. Our experiments demonstrate the feasibility of achieving both gene ablation through NHEJ and gene replacement by HR by using the OPEN ZFN technology directly in mouse zygotes.Zinc finger nucleases (ZFNs) enable precise genome modification in a variety of organisms and cell types. Commercial ZFNs were reported to enhance gene targeting directly in mouse zygotes, whereas similar approaches using publicly available resources have not yet been described. Here we report precise targeted mutagenesis of the mouse genome using Oligomerized Pool Engineering (OPEN) ZFNs. OPEN ZFN can be constructed using publicly available resources and therefore provide an attractive alternative for academic researchers. Two ZFN pairs specific to the mouse genomic locus Mouse lines carrying genes that have been disrupted (knocked-out) or modified (knocked-in) by homologous recombination (HR) are important tools that are widely used in biomedical research. Such lines are generated by gene targeting in mouse embryonic stem (ES) cells and subsequent morula aggregation or blastocyst injection of positive clones to generate chimeric animals FokI http://www.zincfingers.org) Zinc finger nucleases (ZFNs) have been conceived as an alternative means of selectively altering the eukaryotic genome DSBs caused by ZFN activity can be repaired by either error-prone non-homologous end joining (NHEJ), the dominating DNA repair mechanism in most eukaryotes gt(ROSA26)Sor (ROSA26) locus. Our findings demonstrate that OPEN ZFNs can be used to achieve gene ablation through NHEJ and gene targeting by HR directly in mouse zygotes.Here, we tested two OPEN ZFNs designed to target the mouse ROSA26 locus is a \u201csafe harbour\u201d frequently used for site-specific insertion of transgenes by HR. Previous studies have demonstrated the feasibility of gene targeting in the ROSA26 locus by use of commercially available ZFNs ROSA26 locus. Due to constraints in the targeting range of the OPEN system it was, however, not possible to target a ZFN pair directly to the XbaI site in the ROSA26 locus that is frequently used to insert transgenes. Instead, two ZFN pairs were used in our experiments that could mediate DNA cuts in proximity of this XbaI site. These ZFN pairs, 90/91 and 204/205, target the ROSA26 sequence 75 and 403 bp upstream of the XbaI site, respectively . This toxicity also persisted upon co-injecting reduced concentrations (2 ng/\u00b5l) of 204/205 homodimeric mRNAs. Thus, none of the experiments performed with the heterodimeric pair 90/91 and homodimeric pair 204/205 led to any discernable activity in mouse zygotes.Initially, we injected ZFN pairs as FspI digestion and confirmed through sequence analysis , C57BL/6, and CD1 mice were purchased from a commercial breeder . All animals were maintained in temperature- and light-controlled rooms with food and water B6D2F1 or C57BL/6 female mice underwent ovulation induction by intra peritoneal (i.p.) injection of 5 IU pregnant mare's serum gonadotrophin , followed by i.p. injection of 5 IU human chorionic gonadotropin 48 h later. For the recovery of zygotes, the B6D2F1 and C57BL/6 females were mated with the males of the same strain immediately after the administration of hCG. All zygotes were collected from oviducts 24 h after the hCG injection and were then freed from any remaining cumulus cells by a 1\u20132 min treatment of 0.1% hyaluronidase (Sigma) dissolved in M2 medium.2. For micromanipulation the embryos were transferred into M2 medium (Sigma).Mouse embryos were cultured in M16 (Sigma) medium at 37\u00b0C and 5% COAll microinjections were performed using a microinjection system comprised of an inverted microscope equipped with Nomarski optics , set of micromanipulators and a FemtoJet microinjection unit . ZFN mRNAs were injected into the cytoplasm whereas the DNA expression constructs and DNA targeting fragments were injected into the male pronuclei; in experiments where mRNA and DNA were co-injected the RNA DNA mixture was first injected into the male pronucleus and subsequently into the cytoplasm upon the withdrawal of the microinjection capillary. Specific concentrations of injected mRNAs and DNA constructs are compiled in Embryos that survived the microinjection were transferred on the same day into the oviducts of 8\u201316 weeks old pseudopregnant CD-1 females (0.5 days post coitus) that have been mated with sterile TgV males XbaI site within the ROSA26 intron 1 were selected using the previously described OPEN method XbaI-BamHI fragment into either the expression vectors pST1374 or pMLM290/pMLM292 that express homo- or heterodimeric ZFNs, respectively Zinc finger proteins binding target sites 75 and 403 bp upstream of the In vitro mRNA transcription, capping and polyadenylation, was performed using the mMESSAGE mMACHINE T7 Ultra Kit. Prior to injection the mRNAs were purified using the NucAway Spin Columns (Ambion). mRNA quality was verified by denaturing gel electrophoresis and concentration was quantified using spectrophotometry.ROSA26 homology arm and a 1.8 kb 3\u2032 ROSA26 homology arm flanking a central SwaI restriction site. An expression cassette consisting of a 1.6 kb CAG promoter/enhancer followed by the 720 bp EGFP coding region and the 531 bp rabbit beta-globin polyadenylation site was inserted by blunt cloning into the SwaI site to generate targeting vector GTR26-EGFP. To generate targeting vector GTR26-tdT a cassette including the 104 bp Ad2 splice-acceptor followed by a 590 bp triple-STOP-pCMV-IRES fragment, the 1.4 kb tdTomato coding region and the 256 bp TK polyadenylation signal was PCR-amplified from pXLBluescriptII PTS tdTomato (gift of J. Ruiz and K. Rector) using primers AGG GCG CAG TAG TCC AGG GTT TCC and GGC TAT GGC AGG GCT TGC CGC C with Pfu polymerase and cloned into the SwaI site of the GTR26 targeting vector. To generate a linear fragment all GTR26\u2212based targeting vectors were PacI digestion prior to microinjection.Targeting vector GTR26 includes a 1.4 kb 5\u2032 Genomic DNA was extracted from mouse biopsies or fetal tissue using a buffer containing 10 mM Tris-HCl pH 9, 50 mM KCl, 0.45% Nonident p40, 0.45% Tween 20 and Proteinase K. Extracts were subjected to Phenol/Chloroform/Isoamyl alcohol purification, precipitated with Isopropanol, and dissolved in EB buffer (Qiagen).ROSA26 locus, primers RF (GCC GCC CAC CCT CCC CTT CCT C) and RR (CGC CTA CT CCA CTG CAG CTC CC) were used to amplify a 474 bp fragment surrounding the ZFN204/205 target site. 25 \u00b5l of each PCR product were digested with FspI and subsequently resolved on a 2% agarose gel. Samples including undigested PCR fragments were cloned into pGEM-T easy (Promega) for Sanger sequencing.For detecting NHEJ repair at the ROSA26 targeting primers GF (GCC GGG ATC ACT CTC GGC ATG) and RR2 (CAC CAC TGG CTG GCT AAA CTC TGG) amplified the 3\u2032 junction that is specific for the integration of GTR26-204/205-CAG-EGFP into the mouse ROSA26 locus. For Southern Blot analysis 10 \u00b5g of genomic DNA were digested overnight at 37\u00b0C with EcoRI, resolved on a 0.7% agarose gel, and transferred to nylon membranes. Membranes were heat-fixated at 65\u00b0C for 1 h and incubated with prehybdrization solution as described EcoRI/PacI fragment, was generated from the \u201cOrkin\u201d plasmid. The ROSA26 3\u2032 probe, a 615 bp, EcoRI fragment, was generated from plasmid pCRII-Rosa 3\u2032. Hybridization probes were heat denatured, labeled with P32 marked dCTP (Perkin Elmer) using the Ladderman Labelling Kit (Takara). The labeled probe was purified with illustra MicroSpin S-200 HRcolumns and heat-denatured probe in hybridization buffer was added to the membranes for overnight rotation at 65\u00b0C. Membranes were washed three times (5 min) using 2\u00d7 SSC. The membranes were exposed at room temperature for 1\u20133 days and imaged using a Storm 840 phospho-imager (Molecular Dynamics). Digital images of Southern Blots were processed with ImageJ.Targeted integration of donor vectors was assessed by junction PCR and Southern blotting. In case of ZFN204/205 \u2013mediated Figure S1Sequencing of ZFN204/205 cleavage site within ROSA26 locus. (A) Sequencing traces for NHEJ-modified ROSA26 alleles. One representative trace per founder is shown. (B) NHEJ-modified alleles in founder ZGFP112. A deletion of 23 bp around the ZFN204/205 cleavage site could be identified in both ROSA26 alleles in founder ZGFP112. The presence of a C/T SNP (red arrow) 33 bp upstream of the ZFN cleavage site (underlined in red) in this founder enabled the identification of individual ROSA26 alleles. Possible regions of microhomology, which can attract NHEJ repair and increase the likelihood of certain NHEJ repair outcomes are underlined in black.(TIF)Click here for additional data file.Figure S2Sequencing of junction PCR product amplified from a founder carrying a targeted ROSA26 allele. The upper panel shows the parts being sequenced with (1) covering parts of the EGFP open reading frame and the polyadenylation signal and (2) covering the junction of ROSA26 genomic DNA and the 3\u2032 homology arm of integrated targeting construct GTR26_EGFP.(TIF)Click here for additional data file.Table S1Sequences of ZFN target sites. Capital letters denote Zinc finger module binding sequences, bold letters highlight binding to the parallel or antiparallel strand, respectively.(PDF)Click here for additional data file.Text S1Sequences of OPEN Zinc Finger modules used in this study.(PDF)Click here for additional data file."} +{"text": "There is a graded inverse relationship from early stages of chronic kidney disease (CKD) (GFR<75ml/min/1.73m2) with increased cardiovascular risk that is driven by high rates of heart failure and sudden cardiac death. Abnormal left ventricular (LV) deformation is an independent predictor of poor prognosis in end-stage CKD and is thought to reflect the extensive interstitial fibrosis and myocyte disarray documented on LV biopsy in uremic cardiomyopathy (UC). There remains a paucity of data relating to the onset of UC in early stage CKD.Eighteen patients (mean age 51 +/- 15 years) with non-diabetic stage 3 CKD (mean eGFR 50 +/- 16 ml/min/1.73m2 MDRD) were compared with 18 healthy age and sex matched controls. Patients all had well controlled blood pressure (mean 24 hour BP 123 \u00b1 14 / 73 \u00b1 6 mmHg) on an average of 1.1 antihypertensive agents and had no history of cardiovascular disease. Cardiac MRI (1.5T Siemens Avanto) with (SPAMM) tagging was performed. Data was analysed using commercially available software to assess global and transmural LV deformation at SAX basal, mid, apical and long-axis HLA levels.Longitudinal and circumferential strain (S) increased in a graded fashion from epicardium (epi) to mid-wall (mw) to endocardium (endo) at basal, mid and apical ventricular levels in both CKD and controls. Global basal circumferential S and strain rate (SR) were reduced in CKD. Basal circumferential S and SR at epi, mw and endo levels were reduced but there was no transmural difference at mid and apical ventricular levels (Table The earliest systolic abnormalities of UC are altered circumferential basal S and SR and reduced longitudinal function. These parameters might serve as early imaging biomarkers that could be used to activate pharmacological intervention. The circumferential abnormalities differ to those observed with aging, hypertension and HFPEF, suggesting a different pathophysiology and necessitate future studies to address whether focal changes in myocardial interstitial fibrosis are responsible for these changes in deformation.British Heart Foundation FS/11/17/28700, PG /04/109"} +{"text": "Vitamin D deficiency, hypocalcemia and acute immobilization negatively affect bone metabolism and are present in the majority of critically ill patients. Although high bone turnover is highly prevalent in the ICU and might compromise long-term outcome, there are currently no data on fracture risk after critical illness.We assessed bone turnover comparing placebo (P) with a cholecalciferol loading dose (VITD) over a 1-week observation period in critically ill medical patients with vitamin D deficiency (25(OH)D \u226420 ng/ml). Markers of bone and mineral metabolism were analysed. Analyses were repeated at days 3 and 7 after 540,000 IU cholecalciferol or matched placebo were given enterally.Twenty-five critically ill patients with an expected ICU stay of more than 48 hours were included . Bone turnover was accelerated indicating bone loss and further deteriorated during the ICU stay. Calcium levels increased significantly in the vitamin D group only (Table Increased bone resorption is frequent in patients in the medical ICU. Intravenous bisphosphonates have been suggested to mitigate bone loss in patients at risk; however, correction of vitamin D deficiency might be a prerequisite for optimal efficacy in this vulnerable population."} +{"text": "The prognosis of patients with metastatic melanoma is poor and not influenced by systemic therapy with cytotoxic drugs. New targeted agents directed against the RAS-RAF-MEK-ERK pathway show promising activity in early clinical development and particular interest is focused on selective inhibitors of mutant BRAF, which is present in one half of the cases of metastatic melanoma. The majority of patients on early trials of these drugs develop secondary resistance and subsequent disease progression and it is, therefore, critical to understand the underlying escape mechanisms leading to resistance. Metastases develop in 10\u201315% of patients with cutaneous melanoma and there is no evidence from phase-III trials that systemic treatment prolongs survival, nor is there an effective adjuvant therapy after resection of high risk Stage I\u2013III disease (Understanding the genetic heterogeneity in melanoma has become increasingly important with the development of therapies aimed at targeting specific genetic aberrations. Of particular importance in melanoma is the mitogen-activated protein kinase (MAPK) pathway, which normally regulates cell growth, proliferation and differentiation . AberranConstitutively activating somatic missense mutations in BRAF were discovered to be present in a wide variety of human cancers, including papillary thyroid cancer (39\u201369%), cholangiocarcinoma (22%), colorectal cancer (5\u201312%) and borderline ovarian cancer (30%) website.Today more than 75 somatic mutations in the BRAF gene have been identified in melanoma, and all mutations at V600 in Exon 15 constitutively activate BRAF . In BRAFThe frequency of BRAF mutation in primary melanomas ranges from 36 to 45% >wild type (wt) BRAF (IC50 22\u2009nM)>mutant BRAFV599E (IC50 38\u2009nM)), but was also found to have a much broader inhibitory profile, including kinases of the vascular endothelial growth factor receptor 2 and 3 (VEGFR-2 (IC50 90\u2009nM) and VEGFR-3 (IC50 20\u2009nM)), the platelet-derived growth factor receptor-\u03b2(IC50 57\u2009nM), Flt-3 (IC50 58\u2009nM) and c-Kit (IC50 68\u2009nM).The first RAF kinase inhibitor entering early clinical trials was the oral diphenyl urea, sorafenib (Bay 43-9006). Sorafenib has potent RAF isoform kinase inhibitor activities , CRAF (IC50 2.5\u2009nM) and mutant BRAFV600E (IC50 6\u2009nM) kinases and demonstrated potent inhibitory effect in a variety of human xenograft models. The phase-I study of XL281 (NCT00451880) in an unselected patient population showed prolonged disease stabilisation in two patients with BRAFV600E mutant papillary thyroid cancer and some minor responses in nine patients with colorectal, ovarian and non-small-cell lung cancer. Overall XL281 was well tolerated with acceptable rates of grade 3/4 nausea, vomiting and diarrhoea (XL281 (famotidine) is an orally administered inhibitor of the wt BRAF (IC50 100\u2009nM) with high selectivity for the mutant allele (BRAFV600E IC50 31\u2009nM) and has shown selective tumour suppression in mutant BRAF cell lines and xenograft models. In a phase-I clinical trial enriched for patients with mutant BRAF metastatic melanoma 11/16 (68%) achieved partial response (PR) and four patients had minor responses leading to a progression-free survival (PFS) of 8\u20139 months (V600E melanoma patients (n=32) at the MTD dose level of 960\u2009mg twice daily (V600E mutant metastatic melanoma is underway.PLX4032 (RO 5185426) is an ATP competitive, orally administered BRAF inhibitor , wt BRAF (IC50 12\u2009nM) and CRAF (IC50 5\u2009nM) kinases with promising preclinical efficacy data in melanoma. A phase-I trial (NCT00880321) of this oral compound enroled >100 patients with BRAF mutations (primarily melanoma patients) in BRAF mutant melanoma patients, a strategy of tandem MAPK inhibition.Other selective B-RAF inhibitors are currently in preclinical and early clinical development and include GDC-0879, ARQ736 and AZ628, among others.Selective RAF inhibitors display good tolerability with infrequent severe toxicities. Among the common grade 1\u20132 adverse events are skin changes (50\u201370%), fatigue (30\u201350%), diarrhoea (10\u201330%) and nausea (10\u201320%). In the GSK 2118436 phase-I trial, 29% of patients reported mild grade 1/2 headaches and a possible cytokine-related fever in 37% of the patients.Most concerning was the development of cutaneous lesions, including KA and SCC in 15\u201330% of patients on the GSK 2118436 and the PLX4032 studies , selective RAF inhibitors can lead to drastic and impressive early tumour responses in humans, which may be of short duration in some patients. Approximately 20% of patients with mutant BRAF melanoma show no response, and most patients relapse, with a median PFS of 8\u20139 months.V600E mutant cell lines demonstrated increased CRAF signalling via BRAF/CRAF heterodimerisation and resulted in a shift from B-RAF to CRAF dependency. In addition post-transcriptional elevation of CRAF protein levels are thought to decrease intracellular bioavailability of the selective RAF inhibitor AZ628 and subsequent development of resistance of the tumour suppressor gene PTEN . Both, gV600E mutant cells, AKT activation was required for melanoma initiation, demonstrating the inter-dependence of these two pathways in melanoma.AKT has a central role in regulating apoptosis and over-expression (via amplification or mutation) of the isoform AKT3 correlates with tumour progression. Recent preclinical studies have shown that inhibitors of PI3K and AKT3 increased apoptosis and stimulated tumour regression is a molecular chaperone responsible for degradation, stabilisation and activation of a variety of proteins, including HER2, CRAF, BRAF, AKT and MET. In melanoma, HSP90 stabilises CRAF and ARAF and is required for the activity of BRAFanamycin , which pEpigenetic changes via DNA hyper-methylation or changes in DNA remodelling have an important role in cancer development. For example, epigenetic silencing is a frequent mechanism of functional loss of the tumour suppressor gene PTEN in melanoma. Agents, such as the histone deacetylating inhibitors (HDACIs), may restore the tumour supressor function by reversing the effects of epigenetic silencing (Clinical trials of new selective RAF inhibitors are currently recruiting, and two recent \u2018proof of concept\u2019 phase-I studies have shown activity in metastatic melanoma, a disease notoriously resistant to systemic treatment.Despite this progress there are important questions to be addressed in current and planned clinical trials. These include the impact of selective BRAF inhibitors on overall survival and quality of life, the identification of biomarkers of early resistance and relapse, the selection and scheduling of drugs to combine with BRAF inhibitors, and the mechanism and prevention of adverse events, such as SCC."} +{"text": "Genomic instability with frequent DNA copy number alterations is one of the key hallmarks of carcinogenesis. The chromosomal regions with frequent DNA copy number gain and loss in human gastric cancer are still poorly defined. It remains unknown how the DNA copy number variations contributes to the changes of gene expression profiles, especially on the global level.We analyzed DNA copy number alterations in 64 human gastric cancer samples and 8 gastric cancer cell lines using bacterial artificial chromosome (BAC) arrays based comparative genomic hybridization (aCGH). Statistical analysis was applied to correlate previously published gene expression data obtained from cDNA microarrays with corresponding DNA copy number variation data to identify candidate oncogenes and tumor suppressor genes. We found that gastric cancer samples showed recurrent DNA copy number variations, including gains at 5p, 8q, 20p, 20q, and losses at 4q, 9p, 18q, 21q. The most frequent regions of amplification were 20q12 (7/72), 20q12\u201320q13.1 (12/72), 20q13.1\u201320q13.2 (11/72) and 20q13.2\u201320q13.3 (6/72). The most frequent deleted region was 9p21 (8/72). Correlating gene expression array data with aCGH identified 321 candidate oncogenes, which were overexpressed and showed frequent DNA copy number gains; and 12 candidate tumor suppressor genes which were down-regulated and showed frequent DNA copy number losses in human gastric cancers. Three networks of significantly expressed genes in gastric cancer samples were identified by ingenuity pathway analysis..This study provides insight into DNA copy number variations and their contribution to altered gene expression profiles during human gastric cancer development. It provides novel candidate driver oncogenes or tumor suppressor genes for human gastric cancer, useful pathway maps for the future understanding of the molecular pathogenesis of this malignancy, and the construction of new therapeutic targets Gastric cancer is one of the most common malignancies and the second most common cause of cancer related death worldwide Genetic alterations are key events in the development of most tumors, including gastric cancer Expression profiling experiments identified a large numbers of genes which are differentially expressed in normal and tumor tissues. However, most of these genes are likely to be passenger genes which have limited contribution to tumorigenesis. The key challenge has been to identify driver oncogenes or tumor suppressor genes that play important roles during tumor initiation and progression, Genomic DNA copy number variation is an important type of genetic alteration observed in tumor cells, and it contributes to tumor evolution by alterations of the expression of genes within the region Previous studies have reported DNA copy number changes or expression profiles in gastric cancer samples. The studies have also identified common chromosome gains and losses, as well as hundreds of genes that may distinguish tumors from normal tissues The use of archival gastric specimen for the current study was approved by the Ethics Committee of the University of Hong Kong and the Internal Review Boards of University of California, San Francisco.Tumor samples were collected from gastrectomy specimens from the Department of Surgery, Queen Mary Hospital, The University of Hong Kong. Eight gastric cancer cell lines AGS, BGC823, N87, NUGC3, SNU16, SNU5, KATOIII and MNK45 have been described in our previous publications The clinico-pathological parameters of the tumors have been previously published http://cc.ucsf.edu/microarray/). They consisted of 2353 bacterial artificial chromosome (BAC) clones that covered the human genome at 1.5 Mb resolution. For hybridization, 1 \u00b5g of tumor DNA and 1 \u00b5g of gender matched reference DNA was labeled by random priming using Cy3-dCTP and Cy5-dCTP, respectively, with the Bioprime Kit (Invitrogen). Unincorporated fluorescent nucleotides were removed using a Sephadex G-50 column . Sample and reference DNA were mixed with 100 \u00b5g Cot-1, precipitated, and resuspended in hybridization solution. The hybridization solution was denatured for 10 min at 72\u00b0C and then incubated for 1 h at 37\u00b0C. Hybridization was performed for 48\u201372 hrs in a moist chamber on a slow rocking table. Arrays were washed for 10 min in 50% formamide and 2\u00d7SSC at 45\u00b0C, and 10 min in phosphate buffer at room temperature. Slides were mounted in mounting solutions containing 0.3 \u00b5g/ml DAPI. Three single-color intensity images were collected for each array using a charge coupled device camera.Human 1.14 arrays were obtained from the UCSF Cancer Center Array Core . \u201cAnalysis of Copy Errors\u201d (ACE) was performed using a false discovery rate (FDR) of 0.0001 and medium sensitivity. Clustering of all samples was performed in TreeView version 1.60.Samples were categorized based on the experimental results and compared with the clinical data using sip-values for the gene expression and copy number correlations were obtained based on permutation. The labels of expression data were randomly shuffled and the Pearson correlation between gene expression clones and copy number BAC clones were calculated as described previously. This was repeated 1000 times. For each gene expression clone, the p-value was determined as the proportion of times the permutation based correlation was greater than or equal to the observed correlation. The p-values were then corrected for multiple testings by controlling for the false discover rate (FDR) using the Benjamini-Hochberg method R/Bioconductor software, including the CBS program, was used to compute the correlation between copy number change and gene expression. The expression data of the 6688 cDNA clones used in the previous gene expression analysis Functional analysis of the significant genes was performed using Ingenuity Pathway software .To identify DNA copy number alterations in gastric cancers, we applied BAC aCGH to 64 human gastric cancer tissue samples and 8 gastric cancer cell lines. The raw data are available in Next, we analyzed DNA copy number variations in gastric cancer samples with different clinico-pathological features including tumor stage, tumor type, tumor site, tumor differentiation, Helicobacter pylori and EBV infection, as well as the difference between gastric tumor samples and cell lines, . We founHER2, TOP2A, CyclinE, TGFB1, AKT2, MYC) and tumor suppressor genes are found to be located in these loci. Interestingly, a higher number of amplifications and homozygous deletions were identified in the cell lines than in the primary gastric cancer tumor samples.High-level amplifications and homozygous deletions are summarized in In our previous study, we reported the gene expression profile in 90 primary gastric cancer samples compared with their 14 metastatic counterparts and 22 non-neoplastic gastric mucosae, with 6688 cDNA clones showing significant variation across these samples Overall, our data confirm that genomic DNA copy number variations contribute to the regulation of regional gene expression profiles in human gastric cancer samples.To pinpoint candidate oncogenes or tumor suppressor genes, we applied two criteria to the list of 1352 cDNA clones which showed statistically significant correlation between gene expression and corresponding DNA copy number changes. First, we searched for genes that showed 5 more gains than losses or 5 more losses than gains in gastric cancer samples. Next, we matched the gene list with the 3329 cDNA clones that were identified to be differentially expressed between non-tumor gastric tissues and human gastric cancer samples To further illustrate how DNA copy number variations influence gene expression, we analyzed the expression patterns of the 333 candidate oncogenes and tumor suppressor genes in the 62 gastric cancer samples using hierarchical clustering . No assoThe Ingenuity Pathway Analysis (IPA) software was employed to investigate the interactions among the candidate oncogenes or tumor suppressor genes identified by expression array and aCGH. Gene copy number alterations are particularly important as deregulating events in cancer progression. In this study, we analyzed Copy Number Aberrations (CNAs) by array CGH. Frequent gains and losses were identified from the study. Furthermore, chromosomal regions with high levels of amplifications and homozygous deletions were also described. Additionally, correlation between gene expression and DNA CNAs were investigated. Candidate oncogenes and tumor suppressor genes were identified by performing integrated analyses of genome copy number and gene expression. Finally, relationships among these candidate genes and their biological function were described in 3 networks using the Ingenuity pathway analysis. The data support that combining aCGH and gene expression array analysis is a powerful method to identify candidate oncogenes or tumor suppressor genes in human gastric cancer. Consistent with this paper, previous studies have applied similar approaches to identify driver genetic events in other tumor types, such as liver cancer AIB1 and BCAS1. AIB1 (20q12), a steroid receptor co-activator first found amplified in breast and ovarian cancer, is involved in gastric cancer cell proliferation through interaction with nuclear receptors BCAS1 (20q13.2), breast carcinoma amplified sequence 1, is amplified in a variety of tumor types and is associated with more aggressive tumor phenotypes. Up-regulated expression of BCAS1 is significantly correlated with the high level amplification of 20q13 in adenocarcinomas of the gastro-esophageal junction MYC is the most representative one. It is one of the most studied oncogenes, which contributes to the malignancy of many different aggressive and undifferentiated human cancers MYC has been ascribed to its ability to control multiplecellular processes such as cell growth, differentiation, apoptosis, DNA damage response, genomic instability, angiogenesis, and tumor invasiveness In the aCGH analysis, frequent gains and amplifications were detected in gastric cancer samples. Of note, consistent with previous studies ERBB2. Overexpression and/or amplification has been observed in many kinds of cancers, including gastric cancer ERBB2 amplification and overexpression is noted by comparing aCGH and expression array data in our gastric cancer data set Whole genome DNA copy number profile of gastric cancer tissue sample HKG24T. Note that this sample showed the following DNA copy number variations: +3q, +5p, +8q, +13q, +17p, \u22124q, \u221210p and \u221218q. (B) Whole genome DNA copy number profile of gastric cancer cell line N87. Note that this sample showed the following DNA copy number variations: +5p, +8q, +11q, +20q, \u22123p, \u22125q, \u22126p, \u22126q, \u22127q, \u22128p, \u221211p, \u221214q, \u221217p and \u221221q. In addition, it also has amplification at 8q21, 8q24, 11q22 and 17q21.(PDF)Click here for additional data file.Figure S2DNA copy number variations in gastric cancer samples with different tumor differentiation. Data presented are ordered by chromosomal map position of the clones. Lower green bars represent losses or deletions, and the upper red bars represent gains or amplifications. (A) Well & moderate differentiated tumor. (B) Poor differentiated tumor.(PDF)Click here for additional data file.Figure S3DNA copy number variations in gastric cancer samples with different tumor site. Data presented are ordered by chromosomal map position of the clones. Lower green bars represent losses or deletions, and the upper red bars represent gains or amplifications. (A) Tumor site: antrum. (B) Tumor site: body. (C) Tumor site: cardia.(PDF)Click here for additional data file.Figure S4DNA copy number variations in gastric cancer samples with different tumor stage. Data presented are ordered by chromosomal map position of the clones. Lower green bars represent losses or deletions, and the upper red bars represent gains or amplifications. (A) Stage 1 & 2 tumor. (B) Stage 3 & 4 tumor.(PDF)Click here for additional data file.Figure S5DNA copy number variations in gastric cancer samples with different tumor type. Data presented are ordered by chromosomal map position of the clones. Lower green bars represent losses or deletions, and the upper red bars represent gains or amplifications. (A) Tumor type: diffuse type. (B) Tumor type: intestinal type.(PDF)Click here for additional data file.Figure S6DNA copy number variations in gastric cancer tumors or cell lines. Data presented are ordered by chromosomal map position of the clones. Lower green bars represent losses or deletions, and the upper red bars represent gains or amplifications. (A) Tissue samples. (B) Cell lines.(PDF)Click here for additional data file.Figure S7DNA copy number variations in helicobacter pylori negative or positive gastric cancer samples. Data presented are ordered by chromosomal map position of the clones. Lower green bars represent losses or deletions, and the upper red bars represent gains or amplifications. (A) Helicobacter pylori negative. (B) Helicobacter pylori positive.(PDF)Click here for additional data file.Figure S8DNA copy number variations in EB virus negative or positive gastric cancer samples. Data presented are ordered by chromosomal map position of the clones. Lower green bars represent losses or deletions, and the upper red bars represent gains or amplifications. (A) EB virus negative. (B) EB virus positive.(PDF)Click here for additional data file.Figure S9Correlation between DNA copy number variations and global gene expression patterns. Each chromosomal arm was divided into equal number of parts or bins of size 20 Mb and then average pairwise Pearson correlation between gene expression and copy number was calculated for all pairs of binned regions. (A) Box plots of correlation between pairs along the diagonal (cDNA clones with surrounding BAC clones) and pairs off diagonal (cDNA clones with unrelated BAC clones). (B) Heatmap of the average correlation between gene expression and copy number.(PDF)Click here for additional data file.Figure S10Correlations between the clustering pattern and clinical features of gastric tumors. (A) Hierarchical clustering of the patterns of variation in expression of genes in 62 gastric tumors. Each row represents a separate cDNA clone on the microarray and each column represents the expression pattern in a separate tumor sample. The ratio of abundance of transcripts of each gene to its mean abundance across all tissue samples is depicted according to the color scale shown at the bottom. Gray indicates missing or excluded data. The dendrogram at the top of the figure represents the hierarchical clustering of the tumors based on similarity in their pattern of expression of these genes. (B) Clinical features of the 62 gastric tumors.(PDF)Click here for additional data file.Figure S11Top biological functions of candidate genes identified by the ingenuity pathway analysis.(PDF)Click here for additional data file.Figure S12Compared ERBB2 expression values with the corresponding DNA copy number changes in 62 gastric cancer samples.(PDF)Click here for additional data file.Table S1Clinical parameters of 64 gastric cancer samples.(XLS)Click here for additional data file.Table S2The raw data of array-based CGH.(XLS)Click here for additional data file.Table S3DNA copy number variations in gastric cancer samples with different clinical parameters.(XLS)Click here for additional data file.Table S4Loci exhibiting high-level amplification or possible homozygous deletion.(PDF)Click here for additional data file.Table S5Genes show statistical significant correlation between expression values and DNA copy number variations.(XLS)Click here for additional data file.Table S6Candidate oncogenes and tumor suppressor genes identified by correlating expression arrays with aCGH data.(PDF)Click here for additional data file.Table S7Summary of analysis (IPA).(PDF)Click here for additional data file.Table S8The raw data of gene expression array.(XLS)Click here for additional data file."} +{"text": "The quality of water in Viet Nam for handwashing with soap or other disinfectant solutions is unknown. We assessed the risk for hand contamination and compared the efficacy of five hand hygiene methods to remove bacterial contamination in a tertiary Vietnamese hospital.Five fingertip imprints of the dominant hand of 134 healthcare workers (HCWs) were sampled to establish the average bacterial count before and after hand hygiene action using: 1) alcohol-based handrub (ABHR); 2) plain soap and water handwashing with filtered and unfiltered water; 3) 4% chlorhexidine gluconate (CHG) hand antisepsis with filtered and unfiltered water.10. Acinetobacter baumannii, Klebsiella pneumonia, and Staphylococcus aureus were the most commonly isolated bacterial pathogens. Highest average count before hand hygiene was recovered from HCWs without direct patient contact (2.10 \u00b1 0.11 log10). Bacterial counts were markedly reduced after hand hygiene with ABHR and CHG with filtered water . Use of unfiltered water was associated with non-significant reduction in bacterial counts.Average bacterial contamination of hands before hand hygiene was 1.65 logHCWs carry high levels of bacteria on their dominant hand, even without direct patient contact. ABHR as an additional step may overcome the effect of high bacterial counts in unfiltered water when soap and water handwashing is indicated.None declared"} +{"text": "Cross-feedback activation of MAPK and AKT pathways is implicated as a resistance mechanism for cancer therapeutic agents targeting either RAF/MEK or PI3K/AKT/mTOR. It is thus important to have a better understanding of the molecular resistance mechanisms to improve patient survival benefit from these agents. Here we show that BRAFV600E is a negative regulator of the AKT pathway. Expression of BRAFV600E in NIH3T3 cells significantly suppresses MEK inhibitor (RG7167) or mTORC1 inhibitor (rapamycin) induced AKT phosphorylation (pAKT) and downstream signal activation. Treatment-induced pAKT elevation is found in BRAF wild type melanoma cells but not in a subset of melanoma cell lines harboring BRAFV600E. Knock-down of BRAFV600E in these melanoma cells elevates basal pAKT and downstream signals, whereas knock-down of CRAF, MEK1/2 or ERK1/2 or treatment with a BRAF inhibitor have no impact on pAKT. Mechanistically, we show that BRAFV600E interacts with rictor complex (mTORC2) and regulates pAKT through mTORC2. BRAFV600E is identified in mTORC2 after immunoprecipitation of rictor. Knock-down of rictor abrogates BRAFV600E depletion induced pAKT. Knock-down of BRAFV600E enhances cellular enzyme activity of mTORC2. Aberrant activation of AKT pathway by PTEN loss appears to override the negative impact of BRAFV600E on pAKT. Taken together, our findings suggest that in a subset of BRAFV600E melanoma cells, BRAFV600E negatively regulates AKT pathway in a rictor-dependent, MEK/ERK and BRAF kinase-independent manner. Our study reveals a novel molecular mechanism underlying the regulation of feedback loops between the MAPK and AKT pathways. The MAPK and AKT pathway represent the most frequently mutated signaling pathways in human cancers. The high prevalence of dysregulation of these two pathways has provided a rationale for the development of target-based therapeutics for cancer treatment. In malignant melanoma, more than 50% of tumors carry BRAFV600E mutation and 70% have elevated AKT phosphorylation and/or activated mTOR activities BRAF inhibitor vemurafenib has shown remarkable clinical efficacy for the treatment of metastatic or unresectable melanoma with a BRAF V600E mutation In this study, we identified a novel crosstalk mechanism between the two pathways, in which BRAFV600E negatively regulates AKT pathway. This mechanism provides a potential explanation why a limited subset of BRAFV600E melanoma cells are exquisitely sensitive to MEK inhibition and supports the rationale combination of AKT and MEK inhibition as a viable cancer therapeutic strategy.in vitro and in vivo were treated with BRAF inhibitor vemurafenib, it induced the paradoxical elevation of ERK phosphorylation as expected in wild-type BRAF cells AKT phosphorylation is regulated by PDK1 and mTORC2. PDK1 phosphorylates AKT at Thr308 and partially activates AKT To understand how BRAFV600E affects pAKT through mTORC2, we examined a possible interaction of BRAF and mTORC2. When rictor was immunoprecipitated in A375 melanoma cells, BRAF was detected in the rictor complex, in addition to mTOR and sin1 (core components of mTORC2), but not raptor (a component of mTORC1), suggesting that BRAFV600E may interact with mTORC2 directly . Notablyin vitro kinase assay. Using an equal amount of recombinant unphosphorylated AKT as a substrate and adding increasing amounts of rictor immunoprecipitates, we found that mTORC2 purified from NIH3T3 (BRAFV600E) cells had lower enzyme activity compared to that purified from the isogenic paired control NIH3T3 cells , we demonstrated that BRAFV600E not only suppresses MEK inhibitor or rapamycin-induced AKT pathway activation but also negatively regulates basal AKT pathway signaling. Importantly, we show that this regulation is through a direct interaction between BRAFV600E and rictor (mTORC2) complex and independent of BRAF kinase activity and downstream MEK and ERK signals. However, the PTEN mutation was sufficient to override the suppressive role of BRAFV600E as in a subset of melanoma cell lines with concurrent BRAFV600E and PTEN deletion, a BRAFV600E effect on pAKT could not be demonstrated. Our finding provides a new interaction node between the MAPK and AKT pathways and suggests a potential resistance mechanism that may emerge in MEK inhibitor-treated melanoma patients.The detailed molecular mechanism of BRAFV600E modulation of mTORC2 remains to be determined. There is evidence that other RAF family members may mediate signaling transduction in a MEK-independent manner. One example is ASK1, which was shown to be suppressed by CRAF independent of MEK-ERK pathway Our conclusion that BRAFV600E negatively regulates AKT pathway is consistent with a long-held, but poorly understood, observation in melanoma biology. It has been shown that a high percentage of benign nevi harbor BRAV600E, with no detectable AKT phosphorylation. These nevi likely underwent an initial phase of growth upon acquisition of BRAFV600E but eventually succumbed to senescence. In contrast to the benign nevi, a high percentage of primary melanomas have concurrent mutations in BRAF and components of AKT pathway Several previous studies have already suggested that BRAFV600E may negatively regulate AKT pathway. LKB1-AMPK signaling is a negative regulator of AKT pathway and has been shown to be modulated by BRAFV600E in melanomas There is accumulating evidence that MAPK and AKT pathways intersect at multiple levels. RAS activates both pathways through interaction with RAF and PI3K respectively. AKT modulates the MAPK pathway through AKT-dependent phosphorylation of BRAF and CRAF. ERK regulates AKT pathway through restraining either upstream GAP1 or downstream TSC1/2 activities. Our study demonstrates a new regulatory mechanism between MAPK pathway and AKT pathway in that BRAFV600E negatively regulates the AKT pathway through interacting with the rictor (mTORC2) complex. This new regulatory mechanism may provide an explanation of why a subset of melanoma cell lines are exquisitely sensitive to MEK inhibition. Our study adds an additional rationale to support combination therapies with RAF/MEK inhibitors and PI3K/AKT/mTOR inhibitors for the treatment of melanoma patients.All cell lines were purchased from the American Type Culture Collection (ATCC). A375, CHL1, SK-MEL-1, A2058, HT-144, and WM266-4 cell lines were grown in DMEM with 10% FBS. LOX cell line was grown in RPMI with 10% FBS. All cell culture reagents were purchased from Life Technologies. All primary antibodies were purchased from Cell Signaling Technology except the following: anti-BRAF from Santa Cruz Biotechnology, anti-CRAF from BD Biosciences, anti-rictor and anti-raptor from Bethyl Laboratories, anti-sin1 from Millipore and anti-\u03b2-actin from Sigma-Aldrich. Secondary antibodies were purchased from Cell Signaling Technology and Thermo Scientific. The MEK inhibitor RG7167 was synthesized at Chugai Pharmaceuticals and the BRAF inhibitor vemurafenib was synthesized at Roche. Rapamycin was purchased from EMD Chemicals. siGENOME siRNAs and the transfection reagents were purchased from Thermo Scientific and transfected according to manufacturer's protocols. Transient transfection of CHL1 was conducted with X-tremeGENE 9 DNA transfection reagent (Roche). Plasmids expressing wild-type BRAF or BRAFV600E were purchased from OriGene Technologies.Human BRAFV600E cDNA was cloned into pcDNA3-based vector (Promega) and plasmids were transfected into parental NIH3T3 cells (ATCC) for selection. Transfected cells were grown in DMEM with 10% calf serum and 1 mg/ml Geneticin and single clones were selected. Five representative clones were further expanded in DMEM with 10% calf serum and 0.5 mg/ml Geneticin for subsequent experiments.All cells were seeded \u223c30% confluency 1 day before compounds or siRNAs were applied. Cells were harvested at designated times for compound treatment or 18\u201324 hours after siRNA transfection. Most samples were collected under subconfluency (<70%) to minimize cell contact-induced change in AKT phosphorylation.in vitro rictor complex kinase assay, rictor complex was purified from cells lysed with IP buffer. The immunoprecipitates were then captured with protein A agarose beads (Millipore) and incubated in rictor-mTOR kinase buffer with inactive AKT1 (Upstate Biotechnology) as described previously For rictor and raptor complex purification, the immunoprecipitation lysis buffer (IP buffer) was prepared as described previously Western blotting was carried out as previously described in vivo xenograft model was conducted as previously described For anchorage independent growth assay, cells were plated in 0.4% agar in DMEM containing 20% FBS and allowed to grow for 2\u20133 weeks before counting the colony. For low serum/growth factor assay, cells were seeded in the regular medium overnight and subsequently grew in the medium with 0.1% FBS. The CellTiter-Glo Luminescent Cell Viability assay (Promega) was used to assess cell proliferation at designated time points. The Figure S1Western blot analysis of the time course response of AKT, MEK and ERK phosphorylation in NIH3T3 isogenic pair treated with RG7167.(TIF)Click here for additional data file.Figure S2Top left panel: Western blot analysis of MEK and ERK phosphorylation in NIH3T3 isogenic pair. Top right panel: Growth kinetics analysis of NIH3T3 isogenic pair in low serum growth medium in 2D cell culture plates. Lower left panel: Analysis of the anchorage dependency of NIH3T3 isogenic pair in soft agar assay. Lower right panel: Growth kinetics analysis of NIH3T3 isogenic pair in nude mice xenograft model.(TIF)Click here for additional data file.Figure S3Western blot analysis of AKT phosphorylation in A2058 and HT144 melanoma cell lines 24 hours after knock-down of BRAF.(TIF)Click here for additional data file."} +{"text": "The presence and absolute amount of microvascular obstruction (MO) are associated with adverse left ventricular remodelling and prognosis in patients with acute myocardial infarction. MO can be detected with cardiovascular magnetic resonance imaging (CMR) using the technique of first pass perfusion imaging, late contrast enhanced CMR or early contrast enhanced CMR. It has not been shown which standardized definition of hypoenhancement related to the signal of remote myocardium is appropriate to evaluate the presence and the extent of MO on first pass perfusion images.We studied 52 patients with reperfused acute myocardial infarction which showed MO defined as persistent subendocardial hypoenhancement on rest perfusion images. We analyzed rest first-pass perfusion images, early contrast enhanced images and late contrast enhanced images scanned in a 1.5-T magnetic resonance imaging system. The size of MO was measured on rest perfusion images visually and quantitatively using signal intensity (SI) below 2SD and 3SD of remote myocardium as a threshold. The size of MO was also measured visually on early contrast enhanced images and quantitatively on late contrast enhanced images quantitatively defined as the area which did not show SI more than 5 SD above the remote myocardium surrounded by abnormal enhancement.2 and 2.40\u00b11.25 cm2, p=N.S.), whereas those using threshold of \u22123SD of remote myocardium was significantly smaller . MO size analyzed by early contrast enhanced images showed good agreement with visual analysis on perfusion images (ICC=0.881). Mean MO size assessed by late contrast enhanced images was significantly smaller than those on by visual analysis on perfusion images .On rest perfusion images, standardized analysis using the threshold of \u22122SD of remote myocardium showed good agreement with visual analysis (ICC= 0.793). Mean MO size using threshold of \u22122SD of remote myocardium was not significantly different from those by visual assessment (2.22\u00b11.25 cmIn patients with acute, reperfused MI, standard late enhancement images underestimate the size of myocardial areas with microvascular obstruction. Using a cutoff of 2 SD below mean SI of remote myocardium, rest perfusion images should be used to quantify the extent of microvascular obstruction."} +{"text": "Prevalence of colorectal cancer (CRC) in the British Bangladeshi population (BAN) is low compared to British Caucasians (CAU). Genetic background may influence mutations and disease features.We characterized the clinicopathological features of BAN CRCs and interrogated their genomes using mutation profiling and high-density single nucleotide polymorphism (SNP) arrays and compared findings to CAU CRCs.-5) and this difference was not due to Lynch syndrome or the polyposis syndromes. KRAS mutations in BAN microsatellite stable (MSS) CRCs were comparatively rare (5.4%) compared to CAU MSS CRCs , which correlates with the high percentage of mucinous histotype observed (31%) in the BAN samples. No BRAF mutations was seen in our BAN MSS CRCs . Array data revealed similar patterns of gains (chromosome 7 and 8q), losses and LOH in BAN and CAU CRCs. A small deletion on chromosome 16p13.2 involving the alternative splicing factor RBFOX1 only was found in significantly more BAN (50%) than CAU CRCs (15%) cases (p=0.04). Focal deletions targeting the 5\u2019 end of the gene were also identified. Novel RBFOX1 mutations were found in CRC cell lines and tumours; mRNA and protein expression was reduced in tumours.Age of onset of BAN CRC was significantly lower than for CAU patients (p=3.0 x 10KRAS mutations were rare in BAN MSS CRC and a mucinous histotype common. Loss of RBFOX1 may explain the anomalous splicing activity associated with CRC. APC) tumour suppressor gene, MAP patients carry biallelic mutations in the MUTYH oxidative damage repair gene and LS is caused by germline mutations in DNA mismatch repair genes . Colorectal tumours showing RBFOX1 deletions were identified and then clustered using a k-means algorithm. Genomic coordinates are in Human Genome Build hg18.For copy number analysis, SNP data for the tumour samples were pre-processed by regressing Log R Ratio values for the tumour against corresponding values for the paired normal to reduce the effect of array artefacts. The SNP data was then examined for copy number and LOH changes in paired tumour/normal tissue using OncoSNP (version 2.25) [PMID: PMC2965384]. We used the intra-tumour heterogeneity option with 10 EM iterations and sub-sampling window size of 30. Two early-onset BAN and 4 CIN samples were excluded on the basis of poor data quality. For comparisons, copy number and LOH calls from the Illumina Human Hap500 and 610-Quad SNP genotyping data were remapped using nearest neighbour interpolation on to the HumanHap370-Duo probe set. KRAS and BRAF were analysed by direct sequencing (Life Technologies). RBFOX1 exons were screened using high resolution melt curve analysis on xthe ABI 7500 Fast Real Time PCR system (Life Technologies) and analysed using the HRMv2.0.1 software (Life Technologies). Somatic mutations were then validated by bidirectional Sanger sequencing twice and confirmed to be absent in paired normal tissues. Primer sequences are available from authors. Mutations were analysed for the effect of amino acid substitutions on protein structure and function, using Sorts Intolerant From Tolerant (SIFT) at http://sift.bii.a-star.edu.sg/ and Polymorphism Phenotyping (PolyPhen-2) at http://coot.embl.de/PolyPhen/.Mutation hotspots in RBFOX1 (Hs00251554_m1) and thecontrol housekeeping gene HPRT . Immunohistochemistry was carried out using rabbit polyclonal primary antibody against RBFOX1 .Quantitative Real-time polymerase chain reaction (QPCR) was performed using TaqMan assays (Applied Biosystems) against Ethical approval for this study was obtained from the Human Research Ethics Committees of all sites and all patients gave informed consent.The author(s) declare that they have no competing interests.NSe, ASa, NSu, GPE, CL performed the experiments and data analysis. CY, DM, JBC carried out bioinformatics and biostatistics. PG, AG, MT, SA, RF, DP, SD provided tissue samples and clinical data. RF performed analysis for immunohistochemistry. OS, ASi and CL contributed to design the studies, interpretion of the data and wrote the manuscript. All authors read and approved the final manuscript.Table S1. Loss of mismatch repair proteins MLH1, MLH6 and PMS2 in a subset of British Bangladeshis colorectal cancers. No tumour showed loss of MSH2.Click here for fileFigure S1. UCSC Genome Browser showing hemizygous deletions/LOH (Red) and homozygous deletions (Yellow) of RBFOX1 in The Cancer Genome Atlas colorectal adenocarcinoma sample set. Deletions are typically hemizygous loss of the entire gene and focal deletions primarily targeting 5\u2019 end of the gene.Click here for fileTable S2.RBFOX1 mRNA expression in CRC cell lines and paired CRC and normal tissues samples.Click here for fileTable S3. Somatic mutations and single nucleotide polymorphisms found in CRC cell lines and paired CRC and normal tissues samples.Click here for file"} +{"text": "Investigation of pulmonary pathology with computed tomography also allows visualisation of the heart and major vessels. We sought to explore whether clinically relevant cardiac pathology could be identified on computed tomography pulmonary angiograms (CTPA) requested for the exclusion of pulmonary embolism (PE). 100 consecutive CT contrast-enhanced pulmonary angiograms carried out for exclusion of PE at a single centre were assessed retrospectively by two cardiologists.Evidence of PE was reported in 5% of scans. Incidental cardiac findings included: aortic wall calcification (54%), coronary calcification (46%), cardiomegaly (41%), atrial dilatation (18%), mitral annulus calcification (15%), right ventricular dilatation (11%), aortic dilatation (8%) and right ventricular thrombus (1%). Apart from 3 (3%) reports describing cardiomegaly, no other cardiac findings were described in radiologists' reports. Other reported pulmonary abnormalities included: lung nodules (14%), lobar collapse/consolidation (8%), pleural effusion (2%), lobar collapse/consolidation (8%), emphysema (6%) and pleural calcification (4%).CTPAs requested for the exclusion of PE have a high yield of cardiac abnormalities. Although these abnormalities may not have implications for acute clinical management, they may, nevertheless, be important in long-term care. Computed tomography pulmonary angiography (CTPA) has become a first line investigation for suspected pulmonary embolism (PE) -3. AlthoOne hundred consecutive patients aged 64 (19) (mean [SD]) underwent CTPA in the period April to July 2007. The scans, acquired on a non-gated conventional 16-slice CT scanner using bolus tracking, were analysed retrospectively and independently by two cardiologists . Any disagreements in findings were referred to a third cardiologist (HEG) with Level 2 cardiac CT accreditation. These reports were compared with previously issued radiological reports. Both atrial and ventricular dimensions were made in the transverse plane at the mid-ventricular level.Absolute numbers of findings for each type of pathology was converted into a percentage of the total number of scans analysed.As shown in Table According to the retrospective report from cardiologists, 78% of scans had at least one incidental cardiac finding Table . AccordiComputed tomography pulmonary angiography has become a routine investigation for the investigation of acute PE. The British Thoracic Society and European Society of Cardiology guidelines recommend the use of CTPA as the initial investigation method of choice in non-massive PE . In thisAlthough incidental cardiac findings may not be relevant to the immediate clinical management of patients with suspected PE, they may nevertheless influence long-term clinical management. For example, coronary and aortic wall calcification are markers of atherosclerosis ,11. AccoWe conclude that thoracic CT scans frequently yield evidence of incidental cardiovascular pathology. Whilst these findings may not be relevant to immediate clinical management, they may well be clinically relevant in the long-term. Radiologists' reports of thoracic CT scans should therefore include cardiac findings.1. Incidental cardiac findings of significant pathology are common on CT scans of the thorax.2. Reporting of cardiac abnormalities is important as potentially management may be changed as a result3. Cardiac abnormalities are more common than pulmonary emboli on CT scans4. Cardiac failure is associated with cardiomegaly. The presence of cardiomegaly should prompt further cardiac investigation.5. Incidental cardiac findings are usually not reported.The authors declare that they have no competing interests.AH and PWXF analysed the scans together and wrote the manuscript. HEG arbitrated where there was dispute in the findings and contributed to the manuscript. FL inspired the study and as senior author re-wrote parts of the manuscript. All authors have read and approved the final manuscript."} +{"text": "The impact of bundled infection control practices introduced in the Intensive care units (ICU) is analyzed to identify the most effective infection control practice in reducing Ventilator Associated Pneumonia (VAP) rates.VAP data over a period of 24 months (Jan\u201909 to Dec\u201910) in a Tertiary care Indian ICU was retrospectively analyzed. The impact of each intervention of prevention bundle introduced in Dec\u201909 was critically evaluated on VAP rates in the subsequent year 2010.The VAP rates over the entire study period varied between 0 and 30.9/1000 ventilator days. The VAP rate for the year 2009 was 9.72 (22 VAP infections in 2263 ventilator days) which reduced to 3.43 (11 VAP infections in 3202 ventilator days) in the following year 2010 as a result of the interventions. The study showed hand hygiene compliance of the healthcare workers and length of stay of patient as the major risk factors associated with VAP. The most effective intervention strategies analyzed were head of bed elevation (p < 0.0001), sub glottic suction (p < 0.0001), hand hygiene compliance of healthcare workers (p < 0.0001) and daily assessment of weaning and extubation for ventilated patients (p < 0.0001). The other measures like closed endotracheal suctioning (p= 0.257), humidification with heat and moisture exchangers (p=0.091), chlorhexidine mouthcare every 2 hours (p=0.002) and routine drainage of ventilator circuit condensate (p=0.0112) proved to be less significant.Head of bed elevation, hand hygiene compliance of healthcare workers, sub glottic suction and daily assessment of weaning and extubation for ventilated patients contributed as the most effective infection control practices in VAP prevention.None declared."} +{"text": "We demonstrate the feasibility of 3D myocardial perfusion CMR at 3 Tesla against single photon emission computer tomography for the detection and estimate of ischaemic burden and show good agreement between the techniques. This novel technique shows promising use of this method in a small cohort of patients to estimate ischaemic burden for purpose of risk stratification of patients with known or suspected coronary disease.The extent and severity of ischaemia on single photon computed tomography (SPECT) is commonly used to risk-stratify patients with suspected coronary artery disease (CAD). Accurate estimates of ischaemia burden by CMR is limited because conventional two-dimensional myocardial perfusion methods cover the heart in a limited number of non-contiguous sections. More recently, three-dimensional (3D) myocardial perfusion CMR has been proposed to overcome the limitation of spatial coverage but has yet to be validated against SPECT.To compare ischaemia burden on 3D myocardial perfusion CMR with (99m)Tc-tetrofosmin myocardial perfusion SPECT (MPS).Ten consecutive patients with known or suspected CAD who were clinically referred for MPS underwent 3D CMR perfusion. The 3D datasets were analysed by an experienced observer blinded to the MPS data and images were scored for the presence of inducible ischaemia in accordance to the AHA 17 segment model.Semi-quantitative analysis of the ischaemic burden was calculated from the sum stress difference between stress and rest tracer uptake for MPS. The 3D CMR perfusion images were similarly analysed for inducible perfusion abnormalities .3D myocardial perfusion CMR and MPS agreed in nine of the ten patients for the detection of any inducible ischaemia. In one patient, CMR detected ischaemia not seen on MPS. Preliminary diagnostic sensitivity was 100%, specificity 75%, postive predictive value 0.86, negative predictive value 1.0) using SPECT as the gold standard. The mean percentage of inducible ischaemia was 7.3% range 0-22.2%) for CMR and 10.3% for SPECT (range 0-25.0%). Overall there was no significant difference (P=0.47)(Figures 2.2% for 3D myocardial perfusion CMR agrees well with SPECT for the detection of coronary artery disease. Both techniques produced similar volumes of ischaemia in the small cohort studied. 3D myocardial perfusion CMR offers a promising alternative method of detecting ischaemia with the added benefits of improved spatial resolution and avoiding the need for ionising radiation. The method holds promise for the risk stratification of patients with known or suspected CAD.SP is funded by British Heart Foundation fellowship FS/10/62/28409.SP/EN receives research grant support from Philips Healthcare."} +{"text": "Nicotiana tabacum) lines were engineered to ectopically over-express AtMYB90 (PAP2), an R2\u2013R3 Myb gene associated with regulation of anthocyanin production in Arabidopsis thaliana. Independently transformed transgenic lines, Myb27 and Myb237, accumulated large quantities of anthocyanin, generating a dark purple phenotype in nearly all tissues. After self-fertilization, some progeny of the Myb27 line displayed an unexpected pigmentation pattern, with most leaves displaying large sectors of dramatically reduced anthocyanin production. The green-sectored 27Hmo plants were all found to be homozygous for the transgene and, despite a doubled transgene dosage, to have reduced levels of AtMYB90 mRNA. The observed reduction in anthocyanin pigmentation and AtMYB90 mRNA was phenotypically identical to the patterns seen in leaves systemically silenced for the AtMYB90 transgene, and was associated with the presence of AtMYB90-derived siRNA homologous to both strands of a portion of the AtMYB90 transcribed region. Activation of transgene silencing in the Myb27 line was triggered when the 35S::AtMYB90 transgene dosage was doubled, in both Myb27 homozygotes, and in plants containing one copy of each of the independently segregating Myb27 and Myb237 transgene loci. Mapping of sequenced siRNA molecules to the Myb27 TDNA (including flanking tobacco sequences) indicated that the 3\u2032 half of the AtMYB90 transcript is the primary target for siRNA associated silencing in both homozygous Myb27 plants and in systemically silenced tissues. The transgene within the Myb27 line was found to consist of a single, fully intact, copy of the AtMYB90 construct. Silencing appears to initiate in response to elevated levels of transgene mRNA (or an aberrant product thereof) present within a subset of leaf cells, followed by spread of the resulting small RNA to adjacent leaf tissues and subsequent amplification of siRNA production.Transgenic tobacco ( Dramatic variability of transgene expression, including complete silencing of the introduced gene or genes, has been a factor impacting the success of plant genetic engineering since its inception. The observed variability in expression levels of what appear to be identical transgene constructs has been linked to multiple molecular factors such as high transcription levels, alterations to the copy number and orientation of introduced DNA, and the characteristics of closely linked plant genetic material The \u2018gold standard\u2019 for inducing gene silencing in plants involves the production of double-stranded RNA (dsRNA), usually from genetic constructs engineered to produce self-complementary hairpin RNA transcripts Arabidopsis myb transcription factor is consistently and reproducibly associated with doubling of the transgene copy number that occurs within homozygotes (over at least 3 generations). The structurally intact, single copy, 35S::AtMYB90 transgene provides a simple visual indicator of silencing Nicotiana tabacum (Petite Havana SR1) by Agrobacterium tumifaciens transformation (AtMYB90 by producing a darkly pigmented (anthocyanins) phenotype in nearly all plant tissues . All 27Hmo plants were found to reproducibly produce reduced anthocyanin levels relative to their hemizygous (27Hmi) parent or siblings (A. tumefaciens strain containing a hairpin construct (pKOihpMyb) that produces dsRNA homologous to the AtMYB90 coding region were used for subsequent mRNA and siRNA analysis.Quantitative reverse transcription polymerase chain reaction (qRTPCR) confirmed the expected gene-dosage associated increase in e levels . The redA levels . The reli leaves . Except AtMYB90 cDNA (generated by rapid amplification of cDNA ends [RACE]) identified the same transcription start and stop points for mRNA isolated from both silenced (27Hmo) and unsilenced plants and 793 bp adjacent to the TDNA left border (TL) that were shortened by NdeI digestion to one smaller band for each probe to GenBank produced no perfect matches but did identify multiple GenBank entries with highly similar sequences . The matIn summary, the single TDNA insert in the Myb27 line, although lacking \u223c650 bp at the Bar gene end of the original TDNA, was not internally rearranged during insertion into the tobacco genome. The Myb27 TDNA locus is flanked by three non-contiguous tobacco DNA segments that are likely to have been brought into proximity during TDNA integration into the tobacco genome.Agrobactrium infiltration site). These RNA samples were size fractionated (<40 nt), ligated to adaptors, converted to cDNA, and gel purified for deep-sequencing using the SOLiD\u2122 platform Total leaf RNA was isolated from several plant samples: SR1 (wild type tobacco); 27Hmo ; 27Hmi ; 237Hmo ; 27Hmi\u2013237Hmi (silenced leaves); and 27Hmi-SysSil was consistently associated with the presence in leaf tissues of abundant siRNA with homology to both strands of the 3\u2032 end of the AtMYB90 transcribed region. The two independent 27Hmo replicates (O-1 and LL6) showed very similar summation patterns. The 27Hmo profiles, however, differed from the those generated using 27Hmi-SysSil and 27Hmi\u2013237Hmi sequence reads were randomly distributed across the Myb27 TDNA target sequence, Nt-TDNA-Nt . The NtLAtMYB90 transcribed region. A chi-square test using siRNA alignment densities from 27Hmi, 27Hmo and 237Hmo samples, paired with the randomly distributed SR1 sample, indicated that sequence reads from both 27Hmo and 27Hmi samples clustered to the AtMYB90 transcribed region (sequence range 2010\u20133009), relative to the remaining TDNA sequence (including flanking tobacco DNA). While not as obvious as the AtMYB90 clustering seen in the silenced 27Hmo (p<0.0001), clustering of the 27Hmi reads to the AtMYB90 transcribed region was still statistically significant (p\u200a=\u200a0.0019). Although alignments from the 237Hmo sample appear to weakly cluster within the AtMyb90 region, the number of aligned sequence reads was insufficient for statistical confirmation (p\u200a=\u200a0.0659).The few TDNA-derived sequence reads identified within purple, unsilenced, transgenic plant samples (27Hmi and 237Hmo) were similar in number to those seen in the SR1 sample . However, when compared to the remainder of the Nt-TDNA-Nt sequence, mapped 27Hmi siRNA reads were enriched within the AtMYB90 transcribed region were primarily 21 nt long (AtMYB90 homology), the unsilenced 27Hmi sample displayed a size profile similar to that observed in the silenced samples, with 21 nt siRNAs comprising roughly half the total number of reads is skewed towards shorter 19 nt lengths consistent with the SR1 alignments representing a background of random sequence reads. A short representative segment of the AtMYB90 coding region (2659\u20132796), displaying matching 21 nt siRNA sequences (from the 27Hmo-O1 and 27Hmo-LL6 samples) directly aligned with the transgene sequence, is shown in ArabidopsisFor all four silenced plant samples, siRNAs homologous to the nt long , with thof reads . The proAtMyb90 transgene was reproduced in two biological replicates of silenced 27Hmo leaf material. The siRNA profiles for 27SysSil and 27Hmi\u2013237Hmi samples were similar, but not identical, to each other and to the 27Hmo profile. Unsilenced 27Hmi leaf samples contained a limited number of sequence reads homologous to the Myb27 Nt-TDNA-Nt sequence (\u223c0.01% of the number of reads in 27Hmo silenced samples) that cluster to the same segment of the AtMyb90 transcribed region targeted in the silenced samples.In summary, the distribution of siRNAs targeted to the Transgene silencing has always played an important, if poorly understood, role in both research and commercial work involving plant genetic engineering rolBAtMYB90 transgene in silenced versus non-silenced plants.Of the published reports describing homozygosity-linked transgene silencing, most have involved transformed tobacco, have made use of the highly active CaMV-35S viral promoter, and have been described as involving PTGS. It has been generally assumed that the doubling of transgene transcription in homozygous plants resulted in the mRNA levels exceeding a threshold and triggering transgene silencing AtMyb90 transgene that is co-linear with that occurring in the original TDNA construct. This leaves open the question as to what characteristic(s) of the Myb27 transgene are responsible for promoting silencing in 27Hmo plants. The structure of the cloned Myb27 insertion indicates that a deletion of the TDNA right border region, and some reorganization of the flanking tobacco DNA, occurred during integration. However, there is no indication of any duplication and/or inversion of the AtMYB90 coding region. In support of there being a single functional copy of the AtMYB90 transgene in Myb27, a spontaneous single-base nonsense mutation within the AtMYB90 coding region converted the phenotypically dominant Myb27 transgene locus (indicated by anthocyanin accumulation) into a dominant-negative marker that interfered with normal tobacco flower pigment production The genetics of the Myb27 dominant purple phenotype and structure of Myb27 TDNA insertion are consistent with the presence of a single 35S::AtMyb90 coding region make it unlikely that the siRNA is the result of transitive spread of silencing from flanking tobacco DNA. It appears that phenotypically detectable transgene silencing in these purple plant lines is initiated by an alteration to some aspect of the AtMYB90 mRNA associated with the doubled transgene copy number present in 27Hmo or 27Hmi\u2013237Hmi plants.The observed juxtaposition of what appear to be unlinked and repetitive tobacco sequence segments adjacent to the Myb27 TDNA insert indicate that alterations to the tobacco genomic DNA arose during integration. However, there is no indication that these flanking sequences are highly transcribed and/or DICER-LIKE 4 (DCL4), in the processing of siRNA precursor(s). The sequenced samples showed a relatively low abundance of 24 nt siRNAs, the product of DICER-LIKE 3 (DCL3) activity, making it unlikely that the observed 27Hmo transgene silencing results from DNA methylation and transcriptional silencing , ath-TAS4-siRNA81. Work done in Arabidopsis identified the tasiRNA binding site in AtMYB90 mRNA as a target of in vivo cleavage TAS4 transcripts is required for production of ath-TAS4-siRNA81 molecules in ArabidopsisAtMYB90 transcript in an area near the 5\u2032 boundary of the of siRNA cluster or unsilenced (27Hmi) leaves. Also, AtMyb90 mRNA from those tissues could be examined for miR828 or TAS4-siRNA81 directed cleavage of the AtMyb90 transcript using rapid amplification of cDNA Ends (RACE).One possible source of aberrant samples . The possequence . HoweverAtMYB90 coding region indicated in The pZP35SMYB construct (diagrammed in th\u20139th true leaves) were used for mRNA analysis.Protocols for PCR and qRTPCR are as described previously The TDNA borders plus adjacent plant sequences were cloned from 27Hmo total DNA using thermal asymmetric interlaced polymerase chain reaction TAIL-PCR and adapAtMYB90 mRNA was converted to cDNA as described previously AtMYB90 transcripts.The transcription start and polyA addition sites were determined using an invitrogen\u2122 GeneRacer\u2122 kit, following manufactures instructions designed to specifically amplify capped 5\u2032 mRNA ends and insure the correct 5\u2032 transcription start site of mature 35S::Small RNA sequence data from tobacco samples were obtained using the Applied Biosystems SOLiD\u2122 v3.0 or v4.0 platforms. For all samples, 100 \u00b5g of total RNA was enriched for small RNA (\u226440 nt) using the Ambion\u00ae FlashPAGE\u2122 system. Enriched smRNA was cleaned and concentrated using Ambion\u00ae spin columns and all concentrations were determined using the Invitrogen\u00ae Qubit\u00ae fluorometer and the Quant-iT\u2122 RNA assay kit (Molecular Probes\u00ae detection technologies). The Quant-iT\u2122 RNA assay kit was performed according to manufacturers protocols with 3 \u00b5l of the enriched RNA sample. Based on measured concentrations, a total of 50 ng of each of the smRNA enriched samples was used to construct smRNA libraries with the Ambion\u00ae Small RNA Expression Kit (SREK). SREK protocols have been optimized to generate fragments for sequencing representing all small, non-coding RNAs in a sample that range in size from \u223c18\u201340 nt. Briefly, an adapter (Adapter mix A) recognizing the 5\u2032-monophosphate was ligated to each smRNA sample to ensure sequencing of all templates from the 5\u2032 end. Following adapter ligation, cDNA was synthesized by reverse transcription with subsequent RNAseH treatment to remove any RNA/cDNA duplexes and any free adapters or adapter by-products. After conversion into cDNA each library was amplified using one of 10 primer sets included with the SREK kit in order to both append the terminal sequences necessary for SOLiD\u2122 sequencing and to incorporate unique barcodes onto each library. The primer sets used for amplification are identical except for a 6 bp barcode (sequence tag) incorporated into the 3\u2032 primer. Amplification was followed by polyacrylamide gel electrophoresis (PAGE) purification on 6% non-denaturing gels and size selection of 100\u2013150 bp products that were used for attachment to sequencing beads and emulsion PCR (ePCR). All ePCR and bead deposition techniques were done following Applied Biosystems standard protocols. Specifically, a total of 60 pg from each of the purified smRNA libraries was used in two different ePCR reactions. After bead clean up, enrichment of p2 beads, and 3\u2032modification according to standard protocols a total of 70 million beads (35 million from each ePCR) containing the 6 barcoded libraries were deposited into two quadrants of a SOLiD\u2122 sequencing slide. Sequencing was done following a bar-code sequencing run to identify the six different libraries and as suggested for eukaryotic miRNA library sequencing 35 bp (V3.0), or 50 bp (V4.0) reads were collected using the appropriate SOLiD\u2122 fragment library sequencing kit.www.Softgenetics.com), GeneSifter\u2122 from Geospiza, Inc (www.geospiza.com) and BioEdit v7.9.1 (www.mbio.ncsu.edu/bioedit/bioedit.html). Local BLAST alignments used BioEdit v7.9.1 and a 4029 bp target sequence representing the flanking tobacco DNA and TDNA present in the Myb27 line . The area of sequence containing 95% of all the mapped siRNA sequence reads from silenced samples is boxed. The locations of ath-miR828 and ath-TAS4-siR81 homologies are underlined equences . The tra(TIF)Click here for additional data file.Figure S2Reversal of silencing in culture. Leaf segments from green sectors of 27Hmo leaves were surface sterilized and placed on MS media with and without hormones. Callus resulting from induced cellular division did not display silencing of the 35S::AtMYB90 transgene as indicted by the accumulation of high levels of anthocyanin pigments.(TIFF)Click here for additional data file.Figure S3Myb27 Nt-TDNA-Nt PCR from genomic DNA. PCR results from the indicated primer sets (27Hmo) and wild type (SR1) plants. Restriction enzyme digests (PstI) of lambda bacteriophage DNA were used as size markers (sizes are indicated). The PCR product sizes predicted for each primer set are indicated in parentheses.mer sets using to(TIF)Click here for additional data file.Table S1PCR primer sets. The DNA sequence for PCR primer pair sets 1through 7 and for the AtMYB(90) qRTPCR primers are listed, including product size for each product (in base pairs).(DOC)Click here for additional data file."} +{"text": "Mutations in BRAF V600E oncogene (BRAFMT) occurs in 8-15% of colorectal cancer (CRC) patients1. This mutation constitutively activates MAPK signalling, resulting in a proliferative and survival advantage for the tumour cells and oncogenic BRAF status has been linked with poor prognosis2. Despite introduction of the BRAFMT specific inhibitor Vemurafenib in metastatic melanoma3, there is no effective treatment strategy for BRAFMT CRC patients. This study aimed to assess the effectiveness of Ganetespib (HSP90 inhibitor), the multi-kinase inhibitor (CRAF/VEGFR/PDGFR) Sorafenib and the BRAFMT inhibitor Vemurafenib in BRAFMT CRC cell line models.BRAF MT RKO F6-8 (MT/WT) and isogenic wild-type T29 (null/WT) cell lines were used. MTT assays were used to determine IC50 values. Protein expression was determined by Western Blotting. Levels of apoptotic cells were assessed by flow cytometry.The RKO BRAFMT cell line was equally sensitive to Sorafenib and Ganetespib compared to the isogenic BRAFWT clone (IC50 1.4\u00b5M vs. 0.9 \u00b5M and 0.86 nM vs. 0.79nM respectively). Vemurafenib treatment resulted in a strong decrease in MAPK signalling and showed greater specificity towards BRAFMT cells (IC50 of 2.4\u00b5M vs. 3.3 \u00b5M) than BRAFWT cells in cell viability assay. Western blotting showed that neither Sorafenib nor Ganetespib had an observable effect in targeting mutant BRAF. Sorafenib did have some effect at targeting downstream MAPK signalling however Ganetespib had no observable effect on targeting the MAPK pathway in BRAF mutant cell lines.. Vemurafenib targeted BRAF and downstream MEK activation in both mutant-type cell lines but not in the wild-type cell lines. We also found that treatment with Vemurafenib induced activation of the STAT3 survival pathway, highlighting a resistance pathway which may be activated in response to treatment in BRAFMT cell lines.Despite the number of compounds which can target BRAFMT cell lines through inhibition of mutant BRAF directly or indirectly through inhibition of other pathways, our studies have shown that Vemurafenib is the most effective treatment strategy in the cell line model tested. Effects of Ganetespib or Sorafenib treatment did not display specificity towards BRAFMT cells alone nor an effect on mutant BRAF. Vemurafenib has shown selectivity towards BRAFMT cells in which a reduction in MAPK signaling is achieved along with induction of apoptosis. Evidence of the emergence of a potential resistance mechanism via STAT3 following Vemurafenib treatment was also found giving insight into the kinome reprogramming event which takes place following treatment."} +{"text": "CHDF using a polymethylmethacrylate membrane is currently widely applied for nonrenal indications in Japan; this technique is used in the treatment not only of patients with sepsis but also of those with cytokine-induced critical illness such as acute respiratory distress syndrome (ARDS) and pancreatitis. This study aimed to investigate the clinical efficacy of continuous haemodiafiltration using a polymethylmethacrylate membrane haemofilter (PMMA-CHDF) in the treatment of patients with sepsis and ARDS.n = 10), pyelonephritis (n = 5), cholangitis (n = 5), tsutsugami in Scrub typhus disease (n = 1), mamushi snake bite (n = 1), haemophagocytic syndrome (n = 1), antineutrophil cytoplasmic antibody lung disease (n = 1), beriberi heart disease (n = 1) and unknown causes (n = 8)) were enrolled in this study between August 2010 and November 2011. The common cause for ARDS in older patients was aspiration pneumonia. Our study group comprised 15 men and 20 women, aged 35 to 85 years (median age 68 years).Thirty-five patients diagnosed with sepsis belonged to the unknown group. The highest survival rate for patients with ARDS was 95%. Moreover, the urine output significantly increased in the survival group.The present study suggests that cytokine-oriented critical care using PMMA-CHDF might be effective in the treatment of sepsis and ARDS, particularly in the treatment of ARDS associated with aspiration pneumonia in older patients."} +{"text": "The text under the subheading \"Determination of the Median Effective Concentration (EC50)\" is a repeat of the text in the section above it (\"Cell culture and reagents.\"). The correct text is: Determination of the Median Effective Concentration (EC50) For each cell line 1500 cells per well were plated onto 384-well plates 18 hrs before drug additions and then incubated in +37\u02daC, 5 % CO2. EC50 values were determined using CellTiter-Glo cell proliferation assay after four day incubation with the drug dilutions. Six replicate wells were made for each drug dilution point and assay was repeated three times. Luminescence was measured with EnVision 2100 Multilabel platereader . The EC50/IC50 values were calculated using sigmoidal dose response equation in GraphPad Prism version 5.0, GraphPad Software, San Diego, CA, USA."} +{"text": "The bone and cartilage destruction seen inrheumatoid arthritis (RA) is caused by synovial pannus formation, which is characterized by aberrant proliferation of synovial fibroblasts. Inhibition of synovial proliferation has recently been reported to be a promising therapeutic strategy for RA.However, the specific mechanism underlyingdysregulated proliferation of synovial fibroblasts remains unclear.We aimed toidentify and characterize genesthat are involved in the aberrant proliferation of synovial fibroblasts.Microarray analysiswas performed to identifythe genes that had upregulated expression inmice with collagen-induced arthritis (CIA). The effect of candidate genes on the proliferation of synovial fibroblasts was screened using antisense oligodeoxynucleotides and small interfering RNAs (siRNAs).SPACIA1/SAAL1 (synoviocyte proliferation-associated in CIA 1/serum amyloid A-like 1)that was associated with aberrant proliferation of synovial fibroblasts. Immunohistochemicalanalysis indicated that SPACIA1/SAAL1 was strongly expressed in the foot joints of mice with CIA and in the thickened synovial lining of the human RA synovium. Transfection of siRNA targeting SPACIA1/SAAL1into RA synovial fibroblastscould inhibit tumor necrosis factor (TNF)\u03b1-induced proliferation more effectively thanit could inhibit serum-induced proliferation.In addition,the antiproliferative effect of SPACIA1/SAAL1 siRNA was caused byinhibition of cell cycle progression and not by induction of apoptosis.We established transgenic (Tg) mice that overexpressed SPACIA1/SAAL1. These Tg mice did not spontaneously develop arthritis or cancer. However,inducing CIA causedgreatersynovial proliferation and worse diseasein Tg mice thanin wild-type mice.We identified a novel gene named SPACIA1/SAAL1 plays an important role in the aberrant proliferation of synovial fibroblasts under inflammatory conditions."} +{"text": "Introduction. Leptomeningeal carcinomatosis occurs in about 5% of cancer patients. Ocular involvement is a common clinical manifestation and often the presenting clinical feature. Materials and Methods. We report the case of a 52-year old lady with optic neuritis as isolated manifestation of neoplastic meningitis and a review of ocular involvement in neoplastic meningitis. Ocular symptoms were the presenting clinical feature in 34 patients (83%) out of 41 included in our review, the unique manifestation of meningeal carcinomatosis in 3 patients (7%). Visual loss was the presenting clinical manifestation in 17 patients (50%) and was the most common ocular symptom (70%). Other ocular signs were diplopia, ptosis, papilledema, anisocoria, exophthalmos, orbital pain, scotomas, hemianopsia, and nystagmus. Associated clinical symptoms were headache, altered consciousness, meningism, limb weakness, ataxia, dizziness, seizures, and other cranial nerves involvement. All patients except five underwent CSF examination which was normal in 1 patient, pleocytosis was found in 11 patients, increased protein levels were observed in 16 patients, and decreased glucose levels were found in 8 patients. Cytology was positive in 29 patients (76%). Conclusion. Meningeal carcinomatosis should be considered in patients with ocular symptoms even in the absence of other suggestive clinical symptoms. Leptomeningeal carcinomatosis results from dissemination of malignant cells to leptomeninges and can be observed in about 5% of patients with malignancies, but it is likely to become more frequent with the increase of life expectancy in cancer patients . NeoplasA 52-year old lady was referred to our hospital for acute onset, ten days before hospitalization, of left orbital pain and visual loss associated with mild frontal throbbing headache. As symptoms progressed, she underwent ophthalmological evaluation as outpatient six days after symptoms onset, without evidence of significant visual loss and normal fundus oculi examination. Ocular computed tomography performed at that time was unremarkable. On subsequent ophthalmological evaluation five days later a significant visual loss in the left eye was evident with substantially normal fundus oculi examination. She underwent right mastectomy and hormonal therapy (tamoxifen) for infiltrative breast carcinoma in 2007 followed by chemotherapy with AC (cyclophosphamide and doxorubicin) for six months, followed by letrozole. She was on regular followup.On admission in August 2011 neurological examination was normal except for mild anisocoria (left > right) with detectable Markus Gunn sign, left eye visual loss, and global reduction of deep tendon reflexes. She underwent contrast-enhanced cerebral MRI showing nonspecific signal alteration involving frontal subcortical and periventricular white matter and focal contrast enhancement involving the left optic nerve sheath see . VEP revIn about two months she started to complain of left hearing loss. Brainstem auditory-evoked potentials were normal on the left see and audiReferences for this review were identified through a search of PubMed from 1966 to March 8, 2012 with the terms \u201cmeningeal carcinomatosis,\u201d \u201cmeningeal carcinomatosis and review,\u201d \u201cmeningeal carcinomatosis and optic neuritis,\u201d \u201cmeningeal carcinomatosis and ocular manifestations,\u201d and \u201cneoplastic meningitis.\u201d Reference lists of relevant articles were also reviewed. Only articles with ocular manifestation as presenting or associated clinical features were included in our review. We found a total of 34 papers including 33 case reports (34 patients) and 1 case series (7 patients). Information about demographic details along with clinical, instrumental, and radiological findings were recorded and summarized in Ocular symptoms were the presenting clinical feature in 34 out of 41 patients (83%) and were the only manifestation of meningeal carcinomatosis in 3 patients (7%). Visual loss was the presenting clinical manifestation in 17 patients (50%) and was the most common ocular symptom (70%). Visual loss was bilateral in 8 patients and unilateral in 2 patients. Sequential optic nerve involvement was observed in 5 patients. Three patients had sudden onset of blindness. Visual loss was progressive in other six patients. Additional ocular manifestations were diplopia , ptosis (8 patients), papilledema (4), anysocoria (3), exophthalmos (2), orbital pain (2), scotomas (2), hemianopsia (1), and nystagmus (1). Other common symptoms were headache (24 patients), altered consciousness (10 patients), meningism (8 patients), hemiparesis (7), ataxia (7), dizziness (6), and seizures (4). V (4), VII (6), VIII (4), IX and X (2), and XII (1) cranial nerves involvement has also been reported.Twenty-three patients underwent enhanced MRI which was diagnostic in 8 patients (35%). In 7 patients generalized meningeal enhancement was noted, and in 2 patients meningeal enhancement was associated with optic nerves enhancement, which was bilateral in 1 patient and unilateral in the other one. Three patients had cranial nerves enhancement, while isolated optic nerve enhancement was seen in only 1 patient . Other MRI findings included hydrocephalus (1), infarction of basal nuclei (1), and cerebellar enhancement (3). 8 patients had normal MRI findings. 3), increased protein levels were observed in 16 patients (range: 62\u2013334\u2009mg/dL), and decreased glucose levels were found in 8 patients (range: 5\u201343\u2009mg/dL). Citology was positive in 29 patients (76%). The histology of the original tumor was highly variable with solid tumors being more frequently associated with meningeal carcinomatosis. The most common was gastric cancer followed by lymphoma, breast, and lung cancer.All patients except five underwent CSF examination which was normal in 1 patient, and increased cells count was found in 11 patients and melanoma 20%). Meningeal involvement occurs in about 5% of breast cancer patients 0%. Menin. Diagnostic workup includes CSF examination and neuroradiological studies. The presence of malignant cells in the CSF is diagnostic, other supportive features are increased opening pressure, pleocytosis, elevated proteins, and decreased glucose levels. CSF cytology may be negative in 65% of patients on initial examination, but only in 20% of patients on second lumbar puncture .Gadolinium-enhanced MRI is also useful for the diagnosis of meningeal carcinomatosis because enhancement on MRI will reveal any irritation of leptomeninges resulting in cranial nerves or intradural extramedullary enhancement on spinal MRI.Ocular involvement represents a frequent manifestation of meningeal carcinomatosis. The reported frequency of ocular signs may be as high as 90% . The preOcular involvement is a frequent and early clinical manifestation of meningeal carcinomatosis. Moreover clinicians should be aware that ocular involvement may mimic different diseases as shown in our case report, where neoplastic optic nerve involvement was indistinguishable from optic neuritis. Thus meningeal carcinomatosis should be included in the differential diagnosis of diplopia and visual loss in selected patients because diagnosis is often challenging."} +{"text": "CYP2C8 has been associated with ZA-related ONJ in MM (The aminobisphosphonate zoledronic acid (ZA) is the most important antiresorptive agent for the treatment of multiple myeloma (MM)-related bone disease (BD). Osteonecrosis of the jaw (ONJ) is an important complication of ZA-treated MM patients ] or not ONJ. We used the novel Affymetrix DMET\u2122 plus platform , which interrogates 1936 genetic variations in 225 genes associated with phase I\u2013II drug metabolism, disposition and transport . The stuP \u2264 0\u00b705) associated with ONJ occurrence. PPARG (peroxisome proliferator-activated receptor gamma), ABP1{amiloride binding protein 1 [amine oxidase (copper-containing)]}, CHST11 [carbohydrate (chondroitin 4) sulfotransferase 11] and CROT (carnitine O-octanoyltransferase). The different distribution of SNP alleles and genotypes between ONJ patients and control cases are reported in PPARG, showed the strongest association with ONJ. We detected a highly significant (P = 0\u00b70055) differential occurrence of the C/C homozygous (HOM) genotype in 77\u00b77% of ONJ cases (7/9) versus only 10% of controls (1/10) . Moreoves (1/10) . The freDirect nucleotide sequencing was carried out on patient specimens to further confirm the presence of genetic variations, using an Applied Biosystems ABI 3100 Genetic Analyser. We found a concordance rate of 100% between DMET genotyping and sequence analysis .PPARG, at position 12477055 of chromosome 3 (Genome Build 37.1). Although no clinical correlation has been reported for the rs1152003 variant, polymorphisms in PPARG have been associated with increased risk of a variety of diseases . ABP1 encodes a membrane glycoprotein that is expressed in many epithelial and haematopoietic tissues. Moreover, a further three ONJ-associated SNPs map to CHST11, which was recently described as a factor required for proper chondroitin sulfation and cartilage morphogenesis. Expression of the chondroitin sulfotransferase genes is crucial for the correct mammalian bone morphogenesis. Finally, the ONJ-associated rs2097937 maps to CROT, whose protein is involved in the trans-esterification of acyl-CoA molecules.The present study also showed that three SNPs identified in PPARG strongly suggests this genetic variant as candidate biomarker for the identification of MM patients at risk of ONJ if treated with ZA. In fact, the C/C genotype demonstrated an odds ratio of 31\u00b75 for developing ONJ following ZA treatment. Differently from the recent report (Our findings indicate that genetic polymorphisms are involved in the pathogenesis of ONJ in MM patients. The highly significant association of ONJ with the rs1152003 SNP polymorphism in"} +{"text": "Drug reaction with eosinophilia and systemic symptoms (DRESS) is among the most severe forms of drug hypersensitivity . AntiepiWe describe a 55 years old female who developed DRESS with acute liver and kidney failure after being treated with ceftazidime and vancomycine. She was successfully treated with corticosteroids although while tapering prednisone she experienced a recurrence of the skin eruption without any systemic symptoms. She was taken off corticosteroids after 9 months of treatment.Fourteen months after the drug reaction, in vitro tests to identify the causal agent were performed. The lymphocyte transformation test (LTT) showed a marked proliferation to ceftazidime (stimulation index (SI): 17 at 100mcg/mL). CD25 was upregulated on CD4+ (induced expression: 17%) and CD8+ (induced expression: 8%) T cells as shown by flow cytometry when cultured with ceftazidime 50 mcg/mL. IFN-\u03b3 was markedly elevated in the supernatant of peripheral blood mononuclear cells (PBMC) cultured with ceftazidime 50 mcg/mL when compared to the control media (946 vs 13 pg/mL). Vancomycin did not induce a significant response when compared to the control media in the flow cytometry and the IFN-\u03b3 assays.This is the first report of ceftazidime-induced DRESS to be subsequently proven by allergy tests. This case illustrates the importance of considering every susceptible drug as the potential etiologic agent. We also show the usefulness of in vitro tests in their identification."} +{"text": "Interventricular dyssynchrony evaluated with cardiac magnetic resonance is an independent predictor of systolic dysfunction and left ventricular remodeling while intraventricular dyssynchrony is an independent predictor of LV diastolic dysfunction in patients with isolated LBBB. Comprehensive evaluation of both these parameters of dyssynchrony would be useful in selecting patients for cardiac resynchronization therapy.Dyssynchrony in patients with left bundle branch block (LBBB) plays an important role in the development of left ventricular (LV) dilation, systolic dysfunction, progression of heart failure and mortality. Intraventricular dyssynchrony (IntraVD) and interventricular dyssynchrony (InterVD) may have different impacts on myocardial structure and function in patients with LBBB. Our objective was to characterize the independent effects of IntraVD and InterVD on myocardial structure and performance.Thirty-two patients with isolated LBBB were assessed using cardiac magnetic resonance imaging. IntraVD was defined as the difference between the time of maximum systolic wall thickness of the septum and lateral wall of the left ventricle. InterVD was defined as the time difference between the onset of pulmonic and aortic flow. Peak filling rate (PFR) and time to peak filling rate (TPFR) were measured as surrogates of diastolic function.InterVD had a moderate inverse negative correlation with LV ejection fraction (r2=-0.5), and a moderate positive correlation with LV end-diastolic volume index (LVEDVI) (r2=0.4) and LV end-systolic volume index (LVESVI) (r2=0.5). The IntraVD had no significant correlation with LV remodeling and function parameters. Multivariate analysis also demonstrated that InterVD was an independent predictor of left ventricular systolic dysfunction (p<0.0001), increased LVEDVI (p<0.01), and increased LVESVI (P<0.001).In terms of diastolic dysfunction, IntraVD had a moderate positive correlation with THFR (r2= 0.5) and moderate negative correlation with PFR (r2=-0.5). InterVD was not significantly correlated with any diastolic dysfunction parameter. Multivariate analysis confirmed that IntraVD was an independent predictor of left ventricular diastolic dysfunction .InterVD evaluated with CMR is an independent predictor of systolic dysfunction and left ventricular remodeling. IntraVD evaluated with CMR is an independent predictor of LV diastolic dysfunction. A comprehensive evaluation of both these independent measures of dyssynchrony could be helpful in improving the patient selection algorithm for cardiac resynchronization therapy.None."} +{"text": "The accumulation of benzodiazepines (BZD) or their metabolites in critically ill patients may prolong wakening times after its discontinuation increasing their morbidity and leading to the overuse of complementary testing to assess neurological function. Route urine/serum determination of BZD are used in some centers to determine BZD clearance, however the correlation of BZD urine/blood concentration and its clinical action has not been determined in critically ill patients.We aimed to investigate the correlation between the presence of BZD in urine and blood (midazolam-MDZ) with the time of wakening after discontinuation of prolonged intravenous (iv) infusion of MDZ in critical care patients. We also aimed to determine the impact of renal failure (ARF) in the efficacy of those test.Prospective, observational study in a 30-bed adult medical-surgical intensive care unit (ICU) from Oct 2014 to Apr 2015. All patients admitted with GCS>8 who were sedated with iv MDZ infusion >72 hours were followed from sedation discontinuation until complete wakening. Patients with chronic use of BZD previous to ICU admission or treated with oral BZD after MDZ iv discontinuation were excluded. Daily urine qualitative and blood quantitative BZD determinations were taken until the first negative urine BZD result. Renal function was determined using RIFLE scale. Time to wake up was defined as the number of hours from BZD discontinuation to the ability to obey simple commands.A total of 32 patients were included, 8 of whom were excluded due to chronic use of BZD (2[25%]) or treatment with oral BZD after MDZ iv discontinuation (6[75%]). The remaining 24 patients years old, APACHE II 23(SD 8), SAPS 3 40(SD 22) were sedated with 2382.32mg (SD 3402.20) of MDZ during 7(SD 6) days.A strong correlation between MDZ serum levels and urine BZD positivity was found. We also found an statistically significant correlation between the number of days of positive BZD urinary determinations and the time to wake up (p = 0.002), but with a low clinical relevance (R\u00b2 0.392). Discrimination power (ROC) of MDZ serum levels to predict the wakening showed an AUC of 0.71 .Patients with ARF (14 [58%]) had more days of positive BZD urinary determinations compared with the group without ARF (10[41%]), despite receiving lower total doses of MDZ respectively.Qualitative BZD urine determination is not a useful tool to predict wake up times in critically ill patients receiving prolonged MDZ infusions. The determination of serum levels of midazolam can readily predict the wakening time in more than 70% of ICU patients who had received long-term MDZ sedation."} +{"text": "Severity of acute kidney injury (AKI) and fluid overload (FO) are not incorporated into current severity of illness measures and are invisible to the practitioner. The causal relationship and timing between AKI and FO and oxygenation is not clear. The Fluid Overload Kidney Injury Score (FOKIS) is a daily score incorporating subscores for AKI (pRIFLE (creatinine (Cr) and urine output (UOP))), FO / ICU admission weight) >15% in five percentile increments, and exposure to nephrotoxic medications. We previously reported that FOKIS outperforms PRISM in mortality prediction in our pediatric intensive care unit (PICU). We hypothesized that patients with AKI on admission to the PICU developed worse fluid overload and in turn worse oxygenation.We prospectively calculated daily FOKIS scores and subscores in PICU patients. We excluded patients with <7 day stays in order to properly explore the association between timing of AKI and FO and oxygenation by oxygenation index (OI).P = 0.03).Over 18 months, there were 2,830 patients, 436 patients with PICU stay >7 days, 361 patients had complete data for all 7 days. Mortality was 4.5% overall and 11% cohort. A total of 246 patients (68%) had AKI (by FOKISCr or FOKISuop); 205 patients (57%) on day 1, 85 patients (24%) on day 3. Admission or day 3 AKI by either FOKIS subscore (FOKISCr or FOKISuop)) did not predict maxFO or mortality. Increasing total FOKIS score was associated with increasing mortality and increasing OI (Table 1). FOKIS, controlled for PRISM, was an independent predictor of OI (In PICU patients, admission or day 3 AKI alone did not predict maxFO. A composite score that includes both AKI and FO parameters correlated with OI and discriminated survivors from nonsurvivors. FO seems to result from combination of increased fluid exposure with underlying AKI but cannot fully be explained by oliguria in pediatric patients."} +{"text": "Global longitudinal strain (GLS), has shown utility in detecting early subclinical LV dysfunction and has demonstrated significant efficacy in evaluating patients for chemotherapy cardiotoxicity. However, echocardiography is often reliant on operator experience and adequate quality windows for assessment. CMR is not subject to limitations of poor image quality due to its superior endocardial definition. We evaluated CMR derived strain by implementing a tissue tracking algorithm and compared it to GLS by 2D echocardiography in a prospective study of chemotherapy patients.10 patients with breast cancer and receiving cardiotoxic chemotherapy (anthracycline and/or trastuzumab) underwent concurrent transthoracic echocardiography (TTE) for GLS and CMR to assess myocardial function and strain (tissue tracking). 2D TTE was performed by an experienced operator using established techniques (speckle tracking). A novel CMR tissue tracking program (Circle Cardiovascular Imaging Inc) was performed by delineating endocardial and epicardial contours on cine images during diastole in 3 long axis views and automated algorithm was applied to derive strain values for each image. These were averaged, to derive global longitudinal strain by CMR and comparison with GLS by TTE was done using Bland-Altman analysis.The mean age of the women was 46 \u00b1 9.3 (SD) years. Myocardial function was preserved in all patients with mean ejection fraction 71 \u00b1 3% (SD). Bland-Altman analysis of TTE GLS versus CMR GLS reveals a bias of -1.322 \u00b1 1.50 (SD) which is within the 95% limits of agreement . (See Figure CMR derived strain may be used interchangeably with TTE derived GLS. This needs to validated in a larger cohort with a wider range of myocardial dysfunction.N/A."} +{"text": "Staphylococcus aureus (MRSA) have become increasingly prevalent as a community acquired infection. As a result limited treatment options are available with conventional synthetic antibiotics. Bioprospecting natural products with potent antimicrobial activity show promise for developing new drugs against this pathogen. In this study, we have investigated the antimicrobial activity of a purple violet pigment (PVP) from an Antarctic bacterium, Janthinobacterium sp. Ant5-2 on 15 clinical MDR and MRSA strains. The colorimetric resazurin assay was employed to determine the minimum inhibitory concentration (MIC90) of PVP against MDR and MRSA. The MIC90 ranged between 1.57 \u00b5g/mL and 3.13 \u00b5g/mL, which are significantly lower than many antimicrobials tested from natural sources against this pathogen. The spectrophotometrically determined growth analysis and total microscopic counts using Live/dead\u00ae BacLight\u2122 fluorescent stain exhibited a steady decrease in viability of both MDR and MRSA cultures following treatment with PVP at the MIC levels. In silico predictive molecular docking study revealed that PVP could be a DNA-targeting minor groove binding antimicrobial compound. The continued development of novel antimicrobials derived from natural sources with the combination of a suite of conventional antibiotics could stem the rising pandemic of MDR and MRSA along with other deadly microbial pathogens. Multiple drug resistant (MDR) and methicillin-resistant"} +{"text": "The intracellular transport is fundamental for cellular functions, morphogenesis and survival in general including neurons composed of a very long axon and dendrites.We discovered most of the kinesin superfamily motor proteins, KIFs, 45 genes in mammals, elucidated their molecular structures and functional roles by molecular cell biology, molecular genetics, biophysics and structural biology and successfully disclosed the mechanism of intracellular transport fundamental for neuronal functions.In the axon and dendrites KIFs transport their cargos such as synaptic vesicle precursors (KIF1A/KIF1Bbeta), mitochondria , plasma membrane proteins (KIF3/KIF5s), NMDA glutamate receptors (KIF17), AMPA receptors (KIF5s) and mRNA with large protein complex (KIF5s). KIFs mainly recognize and bind their cargoes through adaptor protein complex and release them via phosphorylation of KIFs or GTP hydrolyses of cargo G-protein. Furthermore, using molecular genetics we successfully uncovered that KIFs play significant roles for fundamental physiological phenomena in development and functions of nervous system and that deletion of KIFs causes certain diseases by clarifying followings: 1) KIF1A/KIF1B beta hetero mice serve as a model for neuropathy, 2) KIF3 determines left/right asymmetry by generating cilia and nodal flow in the node of early embryos, 3) KIF17 plays a fundamental role on learning and memory by not only transporting NMDA glutamate receptor in dendrites but also controlling transcription and translation of KIF17 and NMDA receptor mRNAs by enhancing phosphorylated CREB, 4) KIF1A is essential for hippocampal synaptogenesis and learning enhancement in an enriched environment, 5) KIF2A is fundamental for brain wiring by controlling unnecessary elongation of growth cones by depolymerizing microtubules, 6) KIF4 plays a crucial role in the activity-dependent survival of postmitotic neurons in brain development by regulating poly(ADP-ribose) polymerase-1 activity, 7) KIF26A is essential for enteric neuronal development by regulating GDNF-Ret signaling, 8) KIF3 suppresses tumorigenesis by transporting beta-catenin from Golgi to plasma membrane for serving as cell-cell adhesion molecules, inhibiting its accumulation in the nucleus and suppressing hyper proliferation of progenitor cells, 9)KIF5A is essential for GABAa receptor transport and KIF5A deletion causes epilepsy, 10)KIF19A is a microtubule depolymerizing KIF for ciliary length control and its deletion causes female infertility and hydrocephalus based on affected fluid flows, 11)KIF13A transports serotonin receptors to plasma membranes and its deletion causes elevated\u2013anxiety phenotypes.Thus, KIFs play significant roles not only at cellular level, but also in brain function and development. Further, their malfunctions cause diseases such as neuropathy, epilepsy, dementia, elevated anxiety, tumor, megacolon and hydrocephalus."} +{"text": "Alu elements make up the largest family of human mobile elements, numbering 1.1 million copies and comprising 11% of the human genome. As a consequence of evolution and genetic drift, Alu elements of various sequence divergence exist throughout the human genome. Alu/Alu recombination has been shown to cause approximately 0.5% of new human genetic diseases and contribute to extensive genomic structural variation. To begin understanding the molecular mechanisms leading to these rearrangements in mammalian cells, we constructed Alu/Alu recombination reporter cell lines containing Alu elements ranging in sequence divergence from 0%-30% that allow detection of both Alu/Alu recombination and large non-homologous end joining (NHEJ) deletions that range from 1.0 to 1.9 kb in size. Introduction of as little as 0.7% sequence divergence between Alu elements resulted in a significant reduction in recombination, which indicates even small degrees of sequence divergence reduce the efficiency of homology-directed DNA double-strand break (DSB) repair. Further reduction in recombination was observed in a sequence divergence-dependent manner for diverged Alu/Alu recombination constructs with up to 10% sequence divergence. With greater levels of sequence divergence (15%-30%), we observed a significant increase in DSB repair due to a shift from Alu/Alu recombination to variable-length NHEJ which removes sequence between the two Alu elements. This increase in NHEJ deletions depends on the presence of Alu sequence homeology . Analysis of recombination products revealed that Alu/Alu recombination junctions occur more frequently in the first 100 bp of the Alu element within our reporter assay, just as they do in genomic Alu/Alu recombination events. This is the first extensive study characterizing the influence of Alu element sequence divergence on DNA repair, which will inform predictions regarding the effect of Alu element sequence divergence on both the rate and nature of DNA repair events. DNA double-strand breaks (DSBs) are a highly mutagenic form of DNA damage that can be repaired through one of several pathways with varied degrees of sequence preservation. Faithful repair of DSBs often occurs through gene conversion in which a sister chromatid is used as a repair template. Unfaithful repair of DSBs can occur through non-allelic homologous or homeologous recombination, which leads to chromosomal abnormalities such as deletions, duplications, and translocations and has been shown to cause several human genetic diseases. Substrates for these homologous and homeologous events include Alu elements, which are approximately 300 bp elements that comprise ~11% of the human genome. We use a new reporter assay to show that repair of DSBs results in Alu-mediated deletions that resolve through several distinct repair pathways. Either single-strand annealing (SSA) repair or microhomology-mediated end joining occurs \u2018in register\u2019 between two Alu elements when Alu sequence divergence is low. However, with more diverged Alu elements, like those typically found in the human genome, repair of DSBs appears to use the Alu/Alu homeology to direct non-homologous end joining in the general vicinity of the Alu elements. Mutagenic NHEJ repair involving divergent Alu elements may represent a common repair event in primate genomes. DNA double-strand breaks (DSBs) are the most dangerous type of DNA damage due to their tendency to lead to chromosomal rearrangements, a hallmark of tumorigenesis, when they are repaired . One wayRecombination between Alu elements can occur through completely identical (homologous) Alu sequences, but most events involve Alu elements with approximately 20% mismatch relative to one another (homeologous), which reflects the average sequence divergence of proximal elements. For the purposes of this study, we will refer to any DNA repair event that occurs through either homologous or homeologous recombination between two non-allelic Alu elements and generates a single chimeric, Alu element, as Alu/Alu recombination. We consider only genomic deletions in this study because our reporter system only detects DNA repair products that result in a deletion. A number of different pathways can give rise to these Alu/Alu deletions, including single-strand annealing (SSA) repair that may predominate when there are high levels of homology, and mechanisms such as microhomology-mediated end joining (MMEJ) where the microhomology happens to be \u2018in register\u2019 between the two Alu elements, allowing formation of a single chimeric Alu element . This lacis to form a DNA duplex [Repair of a DNA DSB via the SSA pathway occurs between two direct-repeat elements on the same DNA duplex, and results in deletion of the intervening genetic material. It is initiated by nuclease-mediated resection of the DNA ends at the break to generate 3\u2019-single-stranded DNA (ssDNA) overhangs that expose the repeat elements for base pairing (reviewed in ). Rad52 A duplex . The annA duplex . If mismA duplex ,12. AlteA duplex .MMEJ is a form of alternative non-homologous end joining . Like SSIn this study, we characterized the impact of sequence divergence on the repair of DNA DSBs between Alu elements by Alu/Alu recombination and a subset of NHEJ events using an Alu/Alu recombination reporter cassette. For the first time, we have extensively quantified the impact of homeologous sequences on the rate of intrachromosomal Alu/Alu recombination. Using our Alu/Alu recombination reporter system, we observed a striking decrease in the rate of intrachromosomal Alu/Alu recombination when the sequence divergence between the Alu elements used for the repair increased. Additionally, as sequence divergence increased, there was a significant increase in NHEJ repair in the vicinity of the Alu elements that did not result in Alu/Alu recombination. These heterogeneous NHEJ repair events appear to be stimulated by the overall Alu homeology, and therefore represent a new alternative influence on the NHEJ pathway that we have termed homeology-influenced NHEJ (HI-NHEJ). These results will aid in predicting which regions of the human genome may be particularly susceptible to Alu-mediated deletions and will give greater insight into genomic recombination hot spots that lead to human disease.R) gene, a polyadenylation signal (pA), and an I-SceI endonuclease cleavage site to generate a DNA DSB between the two Alu elements (R) gene downstream of the Alu sequences is not initially expressed because it is separated from the EF1\u03b1 promoter by the two Alu sequences, the neoR gene, and a polyadenylation signal promoter upstream of two Alu sequences separated by approximately 1100 bp of sequence encoding a neomycin resistance to direct integration into a FRT site at a single, chromosome-specific location . We stabr) colonies were counted to assess the level of recombination. Each of the three independent cell clones testing Alu/Alu recombination between homologous Alu elements (0%-AARP HEK293FRT cells) gave comparable numbers of puror colonies after I-SceI expression, with an average of 1000 colonies per T75 culture flask gene and green fluorescent protein (eGFP+) to create Alu/Alu recombination blasticidin (AARB) and Alu/Alu recombination eGFP (AARG), respectively. These reporters produced similar results to AARP in HEK293FRT cells (r or blasticidin resistant (blastr) colonies, which indicates recombination resulted in a significant reduction in overall colony number relative to 15%-30% sequence divergence (r colonies with 15%-30% sequence divergence between Alu elements relative to 5%-10% is due to a shift in the type of DNA repair responsible for resolution of the DNA DSB (see below).Because Alu elements of various levels of divergence exist throughout the human genome , we sougelements , which ivergence . This obr colonies, and subjected it to PCR and sequence analysis . Repair via NHEJ was responsible for the vast majority of products that involve two Alu elements with \u226515% sequence divergence (r colonies as sequence divergence between the Alu elements increased from 30% to 75% (scrambled sequences) indicated that there is a sequence homology-driven component to the NHEJ DSB repair that makes use of homeologous Alu sequences to stimulate repair in the vicinity of the Alu elements of repair was due to NHEJ and measured the rate of Alu-mediated deletions . We obseWe next asked whether the MLL Alu elements tested in our study and others displayer colonies from Rad52 knockdown experiments revealed that some (29%) were the result of NHEJ repair between the identical Alu elements pathway might influence the resolution of DNA DSB repair between homeologous Alu elements. In order to address the role of an essential MMR protein, Mlh1, in repair of a DSB flanked by homologous or homeologous Alu sequences, we introduced AARP reporters into the FRT site of HCT116FRT cells (denoted HCT116D cells in ). HCT116vergence , which iRT cells . These rIn comparison with HEK293FRT cells, a greater proportion of repair events recovered from HCT116FRT cells were the result of Alu/Alu recombination in most diverged AARP reporters tested , providicis factors on Alu/Alu recombination in a defined chromatin environment resulted in lower levels of Alu/Alu recombination. Presumably, this inhibitory effect is the result of heteroduplex rejection of the intermediate that forms during Alu/Alu recombination Fig and 6 11,12,43. mlh1-deficient HCT116FRT cells formation may occur at a higher frequency in the vicinity of Alu elements relative to other regions ,49. SomePCR and sequence analysis of DNA repair products from AARP HEK293FRT cells with modest levels of sequence divergence (\u226410%) shows 88% of the Alu/Alu recombination events contained a single-crossover region of homology within the Alu element. As others have suggested , these tThe observation that Alu/Alu recombination occurs preferentially near the 5\u2019 region of Alu elements has been seen previously in naturally occurring examples . AlthougThere are several predicted consequences from our studies regarding patterns of genomic instability due to Alu elements . The firhttp://www.blueheronbio.com) and sub-cloned in pENTR 221. The pAARP/ENTR221 construct consists of two identical Alu elements (Ya5 consensus sequence). Alu1Ya5 is flanked by HindIII and BglII restriction sites, followed by a neomycin resistance gene flanked by NcoI and NdeI restriction sites, a SV40 poly A signal and an I-SceI restriction site. The second Alu element sequence, Alu2Ya5, is flanked by BamHI and EcoRI sites and followed by a puromycin resistance gene flanked by EcoRI and AgeI sites. The Alu/Alu recombination blasticidin (AARB) cassette was constructed by insertion of a blasticidin resistance gene (codon optimized bsr) at the EcoRI and AgeI sites . LR Clonase II plus reactions were performed as per the manufacturer\u2019s protocol.The Alu/Alu recombination puromycin (AARP) cassette was synthesized by Blue Heron Biotechnology, Inc (eI sites . The AluThe following Flp-In cell lines were obtained from Life Technologies: human embryonic kidney cells (HEK 293), Baby Hamster Kidney cells (BHK), Mouse (NIH Swiss) embryonic fibroblast cells (NIH-3T3), and Chinese Hamster Ovary (CHO) cells. HCT116FRT cells were kindly provided by J. Issa (denoted HCT116D in ). The Fl6 cells for HEK293FRT and HCT116FRT and 0.5 \u00d7106 cells for BHK, NIH-3T3 and CHO were seeded in a T75 culture flask 24 h prior to transfection. 48 h after transfection, medium was replaced with fresh medium containing 75 \u03bcg/ml of hygromycin for HEK293FRT cells, 150 \u03bcg/ml of hygromycin for HCT116FRT and 250 \u03bcg/ml hygromycin for all other cell lines. The medium was changed every three days. Twelve days after hygromycin selection, three independent clones were selected for Alu-mediated deletion analysis. Individual hygromycin resistant clones were expanded and maintained in medium containing 500 \u03bcg/ml neomycin (Life Technologies). As a negative control, all Alu/Alu recombination destination vectors were transfected without the pOG44 recombinase vector and no hygromycin resistant clones were detected.To develop stable Alu/Alu recombination cell lines, all Alu/Alu recombination destination vectors were specifically integrated into the FRT recombination site of each parental cell line using 100 ng of destination vector and 900 ng of the FLP recombinase expression plasmid pOG44 using Lipofectamine and Plus reagent as the delivery system according to manufacturer\u2019s protocol (Life Technologies), except for HCT116FRT cells, which used 1 \u03bcg of destination vector and 9 \u03bcg of pOG44. For these experiments, 1.0 \u00d710Primers and probes were designed to the neomycin resistance gene of the AARP construct. The neomycin cassette probe was labeled with 5\u2019 6-FAM and quenched with 3\u2019 BHQ1. Primers and probes were synthesized by IDT . The RPPHindIII and BglII digestion , and similarly, the MLL Alu 2 (Sx) was isolated as a fragment obtained by BamHI and EcoRI digestion. These fragments then were cloned into the same sites within a linearized pAARP/ENTR221 vector and subsequently inserted into pEF5/FRT/V5-DEST using the Gateway system and recombined into cell lines using the Flp-In system as described above (Life Technologies).Sixteen sequence diverged Alu/Alu recombination constructs were generated for this study . All div6 AARP-, AARB- or AARG-HEK 293FRT, 0.5 \u00d7 106 AARP-HCT116FRT, 0.5 \u00d7 106 AARP-BHKFRT, 0.5 \u00d7 106 AARP-NIH-3T3FRT, 0.5 \u00d7 106 AARB-CHOFRT cells were seeded into T75 culture flasks one day before transfection. Cells were transfected the next day with 1.0 \u03bcg I-SceI expression vector crystal violet in 5% (v/v) acetic acid and 2.5% (v/v) isopropanol) to quantify the rate of Alu-mediated deletions. Colonies were counted using a ColCount automated colony counter from Oxford Optronix . Each transfection was performed a minimum of three times in parallel using independent clones in duplicates for each construct and cell type. An additional transfection was performed in parallel to isolate genomic DNA in order to analyze the DNA repair events by PCR and sequencing.To perform Alu-mediated deletion assays, 1.0 \u00d7 10M. Jasin ) or emptr clones was extracted using the DNeasy Blood and Tissue kit . 200\u2013500 ng of gDNA was PCR amplified in a 50 \u03bcL GoTaq reaction as per manufacturer recommendations with the addition of 5% DMSO, using primers EF1-FP and Puro-RP as per manufacturer recommendations.Silencing of Rad52 was performed by transient gene knockdown experiments with small interfering RNAs (siRNA). Rad52 siRNA and non-specific control siRNA were obtained from Santa Cruz Biotechnology . 1x10Protein was extracted from cell lysates using TLB-SDS buffer (0.5% (w/v) SDS, 0.5% (v/v) Triton X-100, 150 mM NaCl, 10 mM EDTA, 50 mM Tris pH 7.2). Protein was separated on a 4\u201312% Tris-Glycine gel (Life Technologies) and blotted using a mouse monoclonal antibody specific to Rad52 (Santa Cruz Biotechnology). Rad52 detection was done using a goat anti-mouse HRP conjugated antibody (Santa Cruz Biotechnology) and visualized and quantified on a ChemiDoc (BioRad).AARG-HEK293FRT cells were collected 72 hours after induction of DSB by I-SceI and eGFP fluorescence was analyzed on an LSR II flow cytometer .P values were considered significant if p <0.05.For most experiments, statistical evaluation was performed by two-tailed, Student\u2019s t-tests using GraphPadPrism 5 software. For statistical analysis between 0%-AARP and diverged AARP constructs, , one-wayS1 FigA) Diagram of the pFRT/lacZeo Flp-In target site vector integrated in the parental cell lines containing FRT sites (Life Technologies). Cells with integrated pFRT/lacZeo were zeocin resistant (zeor). (B) The AARP stable cell lines were created by site-specific recombination at the FRT site. The AARP reporter cassette contains a hygromycin resistance (hygroR) gene used to select a stable cell line upon Flp-recombinase-mediated integration of the cassette. The AARP reporter cassette contains a human elongation factor 1\u03b1 (EF1\u03b1) promoter upstream of a neomycin resistance (neoR) gene. An I-SceI endonuclease cleavage site is positioned between the two Alu elements. Prior to DNA repair, the puromycin resistance (puroR) gene was not expressed due to distance from the EF1\u03b1 promoter and interruption by the neoR gene and ployadenylation (pA) site, which results in puromycin sensitivity (puros). (C) Repair of I-SceI-induced DNA double-strand breaks (DSBs) through Alu/Alu recombination or NHEJ that deletes a sufficient portion of sequence allowed puroR gene expression and selection of repaired AARP cells in media containing puromycin.((TIF)Click here for additional data file.S2 FigA) Schematic of the ddPCR assay. A droplet digital PCR (ddPCR) assay was used to confirm that the AARP cassette was integrated as a single copy into HEK293FRT cells. Two Taqman style primer/probe sets were used (R gene integrated as part of the AARP reporter using a 5\u2019 6-FAM-labelled probe (6-FAM moiety indicated by the blue circle). The second primer/probe set used a HEX-labelled RPP30 probe (HEX moiety indicated by the green circle), which detected a cellular housekeeping gene, RPP30, present as diploid in HEK293FRT cells. (B) Results of the ddPCR assay with parental HEK293FRT cell genomic DNA. During ddPCR, genomic DNA for each sample was partitioned into droplets, which are then thermocycled to the plateau phase of PCR. Each droplet was then measured for FAM and HEX fluorescence and plotted accordingly . Droplets with HEX fluorescence were readily detected from ddPCR with parental HEK293FRT cell genomic DNA, whereas very few droplets displayed FAM fluorescence . These FAM-positive droplets were likely due to non-specific amplification of genomic DNA in these droplets. (C) Results of the ddPCR assay with AARP HEK293FRT cell genomic DNA. Droplets were measured and plotted as in (B). In ddPCR assays with AARP HEK293FRT cell genomic DNA both HEX (RPP30 gene) and FAM (neoR gene) fluorescence was detected. (D) The concentration of the neoR and RPP30 gene DNA in samples tested. The concentration of the neoR and RPP30 gene DNA is shown as calcutated by the QuantaSoft ddPCR software (Bio-Rad) as copies/\u03bcL of input genomic DNA from parental HEK293FRT cells and five different clones of AARP HEK293FRT cells, showing the RPP30 gene exists at roughly twice the copy number of the neoR gene. (E) The copy number of the neor gene relative to the RPP30 gene in samples tested. The relative copy number of the neoR gene with respect to the RPP30 gene is plotted for genomic DNA tested in (D). These results confirm that the neoR gene exists at half the copy number of the RPP30 gene in various AARP HEK293FRT clones.(ere used . The fir(TIF)Click here for additional data file.S3 FigA) baby hamster kidney (BHK) FRT cells and (B) mouse embryonic fibroblast (NIH-3T3) FRT and the AARB reporter was stably integrated into (C) Chinese hamster ovary (CHO) FRT cells Click here for additional data file.S4 FigThe 0% Alu (Alu Ya5 consensus sequence) is the sequence of the Alu elements used in the 0%-AARP cassette. In each of the diverged AARP reporter cassettes, Alu 1 was replaced with a diverged Alu element by subcloning. The sequence alignments of each diverged Alu sequence used is shown with diverged nucleotides relative to 0% Alu marked at the respective positions in red.(TIF)Click here for additional data file.S5 Figr colonies is plotted for these cells, along with 15%-, 20%-, 30%-AARP HEK293FRT cells. Data from at least three independently isolated clones for each AARP cell line are averaged with error bars indicating standard error.Three genomic Alu element pairs as well as two scrambled-sequence Alu element pairs were introduced into AARP and integrated into HEK293FRT cells see . The ave(TIF)Click here for additional data file.S6 FigThe size of the Alu/Alu recombination product is ~1.5 kb while NHEJ repair products have variable sizes (lanes 3 and 7). M = 1 kb DNA ladder (Promega).(TIF)Click here for additional data file.S7 FigR gene has been deleted along with portions of the Alu elements on both sides. The NHEJ repair junctions recovered from isolated puror colonies in the diverged AARP constructs indicated are shown as the size of the variable deletions along with the sequences flanking the deletion on both sides. Deletions ranged in size from 897 base pairs to 1901 base pairs. Alu element sequences are in bold italics. Alu1 or flanking AARP reporter cassette sequence is shown to the left of the NHEJ deletion and Alu2 or flanking AARP reporter cassette sequence is shown to the right of the NHEJ deletion. Underlined sequences indicate a microhomology found at the NHEJ repair junction. The same sequences are shown more schematically in At the top is a schematic of the NHEJ product from our AARP vector system where the Neo(TIF)Click here for additional data file.S8 Figr AARP HEK293FRT colonies tested.A schematic representation of the data presented in (TIF)Click here for additional data file.S9 FigA) The distribution of Alu/Alu recombination junctions for 5%e-AARP (40 bp stretches of sequence homology at the 5\u2019 and 3\u2019 ends of Alu1) HEK293FRT cells was determined by PCR and sequence analysis of DNA repair products from isolated puror colonies. The Alu/Alu recombination product is divided into segments according to the intervals of homology in which the Alu/Alu recombination junction can be mapped. The number of Alu/Alu recombination junctions expected and observed in each interval is shown. While not statistically significant at p<0.05, there is still a slight preference for resolution of repair at the 5\u2019 end of the Alu element. (B) The distribution of Alu/Alu recombination junctions for 5%2M-AARP (contains two stretches of 40 bp of sequence homology in the middle of Alu1) HEK293FRT cells was determined as described in (A). An asterisk (*) marks an interval in which p<0.05 significance as determined by a chi-square test for observed vs. expected. No increase in recombination within the stretches of homology was observed. In fact, we again observe a statistically significant bias for recombination resolution within the first 100 base pairs of the Alu element. (C) The distribution of Alu/Alu recombination junctions for 5%4M-AARP (contains 100 bp of sequence homology in the middle of Alu1) HEK293FRT cells was determined as described in (A). An asterisk (*) marks an interval in which p<0.05 significance as determined by a chi-square test for observed vs. expected. No significant increase in recombination within the stretch of homology was observed. Again, however, we observed a statistically significant increase in recombination repair junctions within the first 100 base pairs of the Alu element compared to the number expected based on a random distribution.The Alu1 sequence in 5%-AARP was modified to include larger stretches of homology to Alu2 (Alu Ya5 consensus sequence) and subsequently inserted into HEK293FRT cells see . (A) The(TIF)Click here for additional data file.S10 FigThe NHEJ repair junctions recovered from isolated puror colonies are shown as the size of the variable deletions along with the sequences flanking the deletion on both sides. Deletions ranged in size from 897 base pairs to 1901 base pairs. Alu element sequences are in bold italics. Alu1 or flanking AARP reporter cassette sequence is shown to the left of the NHEJ deletion and Alu2 or flanking AARP reporter cassette sequence is shown to the right of the NHEJ deletion. Underlined sequences indicate a microhomology found at the NHEJ repair junction.(TIF)Click here for additional data file.S11 FigBlack bars above the sequence represent the interval containing the Alu/Alu recombination junctions reported in this study. Similarly, blue bars represent those reported by and oran(TIF)Click here for additional data file.S12 Figr colonies is plotted for the indicated AARP HCT116FRT cells tested. Data from at least three independent experiments using at least three independently isolated clones for each AARP cell line are averaged with error bars indicating standard error.Stably integrated AARP HCT116 cell lines were generated using the Flp-In system see and tran(TIF)Click here for additional data file.S13 Figr colonies for AARP HEK293FRT cells (r colony formation within the 15\u201330% Alu element sequence divergence as observed with HEK293FRT cells.The average number of puroRT cells and AARPRT cells is shown(TIF)Click here for additional data file.S14 Figr colonies is shown.The percentage of Alu/Alu recombination or NHEJ as determined by PCR and sequence analysis of DNA repair products from isolated puro(TIF)Click here for additional data file.S15 Fig(A) A schematic of the Alu/Alu recombination product as shown in (B) The distribution of Alu/Alu recombination junctions in diverged AARP HCT116FRT cells as determined by PCR and sequence analysis of DNA repair products from isolated puror colonies. The Alu/Alu recombination product is divided into three segments of equal length (100 bp), which each contain the same extent of sequence divergence. The number of Alu/Alu recombination junctions expected and observed in each 100 bp interval is shown.(TIF)Click here for additional data file.S16 Fig(A) Schematic showing the typical Alu/Alu recombination product observed. The majority of intact Alu elements show a single conversion location within the Alu element, which implicates MMEJ as the repair pathway used for the majority of Alu/Alu recombination events between diverged Alu elements. (B) Schematics showing the complex Alu element complex chimeras generated in a minority of Alu/Alu recombination events in 5%-, 10%-, and 15%-AARP HEK293FRT cells. Multiple conversion locations can be observed in these Alu/Alu recombination events, which implicates repair of mismatched tracts within a SSA heteroduplex intermediate to generate \u201cpatches\u201d of Alu1 and Alu2 in each AARP clone characterized.(TIF)Click here for additional data file.S17 Fig(A) 5%-AARP, (B) 10%-AARP, and (C) 15%-AARP HEK293FRT cells.Sequence alignments of the complex chimeras generated during Alu/Alu recombination events described schematically in (TIF)Click here for additional data file.S18 Fig(A) A schematic of the Alu/Alu recombination product as shown in (B) The distribution of Alu/Alu recombination junctions in indicated AARP HEK293FRT cells treated with Rad52 siRNAs as determined by PCR and sequence analysis of DNA repair products from isolated puror colonies. The Alu/Alu recombination product is divided into three segments of equal length (100 bp), which each contain the same extent of sequence divergence. The number of Alu/Alu recombination junctions expected and observed in each 100 bp interval is shown. An asterisk (*) marks an interval in which p<0.05 significance as determined by a chi-square test for observed vs. expected.(TIF)Click here for additional data file.S19 FiglacZeo vector at Chr 12 in HEK293FRT cells are shown as determined by whole genome sequencing with Illumina Next Generation Sequencing.Both sequence junctions of the integration of the pFRT/(TIF)Click here for additional data file.S1 TableThe names incorporate the percent divergence between Alu2 (Ya5 consensus) and the variant Alu1 placed in most of the vectors. Most of the actual Alu1 sequences for these vectors are shown in (DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 TableThe results of this experiment are shown in (DOCX)Click here for additional data file.S4 Table6 cells are transfected with control (No DSBs) or I-SceI expression plasmid. Because no replating occurs before colony formation, the number of repair events measured by puror colonies directly correlates with the repair frequency and it has been adjusted to frequency per 100,000 cells for more direct comparison with other studies and given with standard error.In each experiment 10(DOCX)Click here for additional data file."} +{"text": "Errors were made during the preparation of this manuscript that deviate from accepted scientific norms. Specifically, (1) one figure is flipped horizontally in the control panel of two figures representing the same experiment and S1A;The authors provide the correct versions of Figs The subsection \u2018Cloning of amiRs.\u2019 in the Materials and Methods section should read:in silico with flanking KpnI and HindIII sites and synthesized at GeneScript (USA). amiRs were subcloned into EF1:BJ36 using KpnI and HindIII and later into binary vectors using NotI/ScaI and blue white selection. amiRMpIAA9 was cloned into HART (a binary vector conferring Hygromicin Resistance in plants) and amiRMpIAA7 was cloned into KART (Binary Vector conferring G418 Resistance).Cloning of amiRs. amiRs 268 bp in length were designed S8 Fig(A) Phenotypes of several independent lines constitutively expressing amiRMpIAA9. (B) Semi-quantitative RT-PCR showing transgene (amiRMpIAA9) and full-length target (MpIAA) levels in thallus tissues. (C) Transcript levels relative to EF1-alfa control. Lines with the weakest phenotype (line 1 and 4 as seen in A) have the lowest amiR transgene levels.(JPG)Click here for additional data file."} +{"text": "The prognostic value of intra-tumoral cytotoxic T cells has been demonstrated in multiple tumor types. However, immune infiltrate heterogeneity can lead to potentially significant misrepresentations of marker prevalence in routine histological sections. We examined step sections of breast and colorectal cancer samples for CD8+ T cell prevalence by standard chromogenic immunohistochemistry to determine marker variability and inform practice of T cell biomarker assessment in formalin-fixed, paraffin-embedded (FFPE) tissue samples.Serial sections were cut at approximately 25 um intervals from 13 primary colorectal and 12 breast carcinoma FFPE samples until blocks were exhausted and stained for CD8 using DAB-based chromogenic immunohistochemistry. Stained sections were digitally imaged and CD8+ lymphocytes within defined regions of interest (ROI) including the tumor and surrounding stroma were enumerated. Using a linear model/ANOVA framework, statistical analyses of CD8+ cell count variance between patients as well as between levels within a patient sample were performed. Means, medians, and dispersion values of percent CD8 positive cells were calculated. Similarity of CD8 prevalence between slices of a given sample was measured by percent of variability accounted for by slice as a factor in a linear model and an intra-class correlation coefficient (ICC) ranging from zero (no similarity) to 1 .The mean and median CD8+ cell percentages varied between breast ductal adenocarcinoma and colonic adenocarcinoma samples and were highest in medullary breast carcinoma . However, minimal variation in cytotoxic T cell counts between levels within any given tumor block was observed. Percent variability (%) in CD8 counts between step sections from colonic adenocarcinoma (0.1%), medullary breast carcinoma (0.1%) and ductal adenocarcinoma (0.1%) blocks were minor. ICC calculated for percent CD8 positive cells between levels of a sample was 0.99 for both tumor types. Resampling-based methods show CD8 as reliable marker for classifying patients as CD8-positive or negative over a range of cut-offs.Our results show that CD8+ T cell distribution is highly homogeneous within a standard tissue sample in both colorectal and breast carcinomas. As such, cytotoxic CD8 T cell prevalence by immunohistochemistry on a single level can be considered representative of cytotoxic CD8 T cell infiltration for the entire tumor section within the block. These findings support the technical validity of biomarker strategies relying on CD8 immunohistochemistry in FFPE tumors."} +{"text": "Trifolium subterraneum (subterranean clover) and Cicer arietinum (chickpea) and compare these to a partial viral genome sequence obtained from Macroptilium lathyroides (phasey bean). We propose the name phasey bean mild yellows virus for this novel polerovirus.We present here the complete genome sequences of a novel polerovirus from Luteoviridae consists of three separate genera, Luteovirus, Enamovirus, and Polerovirus. Each contains either 5 or 6 open reading frames (ORFs), but only four of these are common to all genera (\u2013The family l genera \u20133. One oTrifolium subterraneum (subterranean clover) collected in 2013 from Esperance, Western Australia (isolate ESPCL15), and one sample of Cicer arietinum (chickpea) collected in 2012 from Liverpool Plains, New South Wales, Australia (isolate NSWCP15), were extracted for total RNA using the Spectrum plant total RNA kit . The total RNA extracts were sent to the Australian Genome Research Facility (AGRF) for library preparation and 100-bp paired-end sequencing on an Illumina HiSeq 2000. The reads from each sample were assembled and the genomes annotated using CLC Genomics Workbench 6.5 (CLC bio) and Geneious 6.1.6 (Biomatters), as described by Kehoe et al. with mild yellowing symptoms were collected in 2011 from Emerald, Queensland, Australia. A reference sample (isolate 2805) was lodged in the Queensland Government plant virus collection. From this, total RNA was extracted using a BioSprint 15 plant DNA kit (Qiagen), as per the manufacturer\u2019s instructions, except that RNase A was not used. PCR products were amplified using previously described conditions and cycling parameters , KT963000 (isolate ESPCL15), and KT906372 (isolate 2805).The sequences were deposited in GenBank with the accession numbers"} +{"text": "Pichia pastoris is a methylotrophic species of yeast, which means that it can grow on methanol as its sole carbon and energy source [Pichia expression system has several advantages: short length of the oligosaccharide chains added to proteins, correct folding and very high cell densities [P. pastoris vectors are designed for homologous integration into either AOX I locus, one of the two homologous AOX I genes present in this species or his4 locus. Gene insertion events at the AOX I (GS115) loci arise from a single crossover events between the loci and any of the three AOX I regions in the vector, the AOX I promoter, the AOX I transcription termination region (TT), or sequences even further downstream of AOX I (3\u2032 AOX I) [y source . The Picensities . P. pastof AOX I \u2032 AOX I [of AOX I \u2032 AOX I [P pastoris GS115 (phenotype His+/Muts) with pPG Vector (pPIC9/ glucoamylase cDNA of Aspegillus awamori) integrad in the AOX locus were transformad with pPAmy vector (pPIC9 vector/\u03b1-amylase gene of Bacillus licheniformis), both previouly contructed in our laboratory. The pPAmy was linearizad by digesting with SacI enzyme to HIS4 locus integration. Transformation was carried out by electroporation of freshly prepared competent cells. The electroporated cells were recoved in 1 M sorbitol and spread onto MM agar plates containg starch 1 % (p/v) to identify positive clones. Transformants with larger halos and clear were considered as positive recombinant strains expressing tow enzymes in comparition the control strain (Strain previously constructed). Positive strains were then inoculated in 50 ml BMGY at 28 C for 48 h in shaking flasks. The cells were harvested by centrifugation and then grown in 25-ml BMMY with methanol induction at 28 C for 120 h. The supernatants were collected by centrifugation and subjected to enzyme activity assay by Fuwa and DNS methods. Extracellular culture supernatant samples were used to proteins detection by SDS-PAGE 10%.Recombinants clones A total of 744 transformants were screened on MM plates. Eight positive transformants with the highest halos amylase activities were analyzed by SDS-PAGE. The gel electrophoresis revealed expression two protein. One protein band with molecular weight of 116 kDa and another with 58 kDa, correponding to glucoamylase and \u03b1-amylase respectively. When we compared the protein pattern of secreted enzymes is agree with protein pattern observed in control supernadant, previously studed in our laboratory. However, in preliminary analyse, none enzymatic activety was detectad in enzymatc assay by Fuwa and DNS methods. However, new transformations and integrations strategies has been performing by study group."} +{"text": "Kawasaki Disease is a generalised systemic vasculitis involving blood vessels throughout the body, although the coronary arteries virtually always are involved. Previous evidence from echocardiographic, Single Photon Emission Computed Tomography (SPECT) and Positron Emission Tomography (PET) studies of myocardial blood flow and flow reserve in KD patients revealed reduced hyperemic flows and flow reserve, suggestive of an impaired vasodilatory capacity. The aim of the study was to determine whether myocardial blood flow and flow reserve derived from perfusion Cardiac Magnetic Resonance Imaging (CMR), is impaired in children with a previous history of KD.Patients with known chronic KD and coronary involvement (mean age: 10.2 \u00b1 7.2 years/mean duration from KD acute illness to CMR exam: 4.95 \u00b1 5.6 years) underwent functional imaging, first-pass stress/rest perfusion, Late Gadolinium Enhancement (LGE) and Magnetic Resonance Angiography (MRA). All exams were analysed visually and quantitatively. Perfusion myocardial reserve was assessed by semi- quantitative methods (Mean Perfusion Reserve Index- MPRI). Mean per patient and segmental results were derived. Results were correlated to the epicardial vessels patency.Fourteen patients with known chronic KD underwent the CMR protocol (mean age: 10.2 \u00b1 7.2 years/mean duration from KD acute illness to CMR exam: 4.95 \u00b1 5.6 years). 5/14 patients presented with persisting epicardial lesions in MRA. 1/14 patient presented inducible perfusion defect. All patients had significantly impaired mean perfusion reserve index compared to normal adult subjects . Perfusion reserve impairment concerned all segments analysed, irrespective of the coronary artery status involved. There was no statistical difference in mean MPRI between patients without and with persisting CALs . comprehensive Biomedical Research Centre award to Guy's & St Thomas' NHS Foundation Trust in partnership with King's College London and King's College Hospital NHS Foundation Trust. The Division of Imaging Sciences receives also support as the Centre of Excellence in Medical Engineering as well as the BHF Centre of Excellence (British Heart Foundation award RE/08/03). Dr Bratis acknowledge receiving training grant by the Hellenic Society of Cardiology. The rest of the authors have no financial activities related to the present article to disclose."} +{"text": "The figures in the PDF version of this paper are of lower quality than intended. The publisher apologizes for the error. Please see the original figures provided by the authors here, as Supporting Information.S1 FigTribes with ambiguous placement are highlighted as follows: most ancient Tanganyika tribes (continuous line); Eretmodini (dashed line); haplochromine lineages (dotted line). Numbers in brackets correspond to the number of individuals.(TIF)Click here for additional data file.S2 FigThe topology of the best scoring ML tree is based on ND2 sequences (1010bp). Numbers at nodes refer to bootstrap-values (BS 500 replicates), 100% BS support is indicated by filled, black circles. Significant deviations from the nc-based NJ-topology are highlighted in red.(TIF)Click here for additional data file.S3 FigThe NJ consensus topology is based on Jaccard\u2019s distances [62] of 3312 nc loci. Nodes affected by homoplasious effects are designated with letters A-M and indicated by open, red circles. Numbers at nodes refer to bootstrap-values (BS 500 replicates) and a 100% BS support is indicated by filled, black circles. Geographic distribution of taxa is depicted vertically on the right and colour shaded in the tree . Major effects detected with HET and inference of cyto-nuclear discordances are delineated in coloured boxes on the left and correspond to those in Fig 6 and Fig 8. Arrows and coloured branches point to clades which especially introduce homoplasy in the dataset. Strong alternative signal (BS support >30) is denoted with dotted lines on the left.(TIF)Click here for additional data file.S4 FigConsensus topology with branch support values depicted at nodes, dots correspond to 100% Bayesian Posterior Probability.(TIF)Click here for additional data file.S5 Fig50% majority rule consensus topology of 1000 BS replicates. BS support is depicted at respective nodes, dots correspond to 100 BS support.(TIF)Click here for additional data file.S6 FigFor all affected nodes the bootstrap (BS) support in the consensus AFLP topology as well as BS support after the influential removals is shown and removed taxa are specified. Node IDs correspond to those in the nc NJ-consensus phylogeny . Single effects were grouped according to four major effects and are represented by different colours. Potentially artificial BS increases due to \u2018support carryover\u2019 (see methods) are highlighted with red frames.(TIF)Click here for additional data file.S7 FigTo infer phylogenetic relationships in an altered variance space, major sistergroups of ingroup taxa according to the consensus NJ topology were stepwise eliminated. NMDS plots are based on Jaccard\u00b4s distances [62] of nc (AFLP) data. Kruskall\u2019s stress values as well as the corresponding 1% cutoff values [74] are given for each projection.(TIF)Click here for additional data file.S8 FigThe NeighborNet network topology is based on Jaccard\u2019s distances [62]. Specimens of distinct lineages are grouped and informal groups used in this work are depicted in different colours. Funnels highlight conflicting signal corresponding to the four major effects detected with HET and inference of cyto-nuclear discordances and are coloured according to Fig 3 and Fig 6.(TIF)Click here for additional data file."} +{"text": "The presence of myocardial late gadolinium enhancement (LGE) in patients with sarcoidosis is a powerful predictor of major adverse events . In this study, we aim to determine if the burden of LGE, left ventricular (LV) and right ventricular (RV) end-diastolic and end-systolic volume index (EDVi & ESVi) and ejection fraction (EF) can be used to improve risk stratification in patients with cardiac sarcoidosis (CS).We identified 25 consecutive subjects with a diagnosis of CS (biopsy-proven extra-cardiac sarcoidosis and presence of LGE on cardiovascular magnetic resonance). Imaging was performed on a 1.5T scanner. Short axis cines spanning the entire LV were acquired to determine LV and RV volumes. Short axis slices were also obtained 10 minutes after contrast (gadodiamide 0.15 mmol/kg) using a phase sensitive inversion recovery reconstruction. LGE was identified on each slice as regions with signal intensity (SI) > 5 standard deviations above the mean SI of normal remote myocardium. The total amount of LGE as a percentage of LV mass (%LGE) was determined using Diagnosoft Virtue software were female. Mean follow up was 39 \u00b1 15 months. Six (24%) of patients had MAE. There was a significant difference in %LGE, RVEF and RVESVi but not LVEF, LVESVi, LVEDVi or RVEDVi between those who had MAE and those who did not (Table The burden of LGE and RV size and function (not LV size and function) further improve prediction of death and significant ventricular arrhythmia in patients with cardiac sarcoidosisNone."} +{"text": "Hypoplastic left heart syndrome (HLHS) is a fatal congenital heart disease in which the left side of the heart is underdeveloped, impairing the systemic circulation. Underdeveloped left ventricle exerts biomechanical stress on the right ventricle that can progress into heart failure. Genome-wide transcriptome changes have been identified at early stages in the right ventricle (RV) of infants with HLHS, although the molecular mechanisms remain unknown. Here, we demonstrate that the RNA binding protein Rbfox2, which is mutated in HLHS patients, is a contributor to transcriptome changes in HLHS patient RVs. Our results indicate that majority of transcripts differentially expressed in HLHS patient hearts have validated Rbfox2 binding sites. We show that Rbfox2 regulates mRNA levels of targets with 3\u2019UTR binding sites contributing to aberrant gene expression in HLHS patients. Strikingly, the Rbfox2 nonsense mutation identified in HLHS patients truncates the protein, impairs its subcellular distribution and adversely affects its function in RNA metabolism. Overall, our findings uncover a novel role for Rbfox2 in controlling transcriptome in HLHS. Hypoplastic left heart syndrome (HLHS) is one of the least understood, fatal congenital heart diseases that affects 1 in 4,344 newborns1145Although genome-wide gene expression changes were identified in HLHS patient RVsde novo detrimental Rbfox2 mutations significantly associated with HLHS phenotype8101114The RNA binding protein Rbfox2 has been recently identified as a major risk allele for HLHSIn this study, we provide evidence that Rbfox2 has a role in HLHS pathogenesis. We demonstrate that HLHS patient specific mutation in Rbfox2 induces a dramatic change in Rbfox2 protein subcellular localization and affects its function in RNA splicing. In the future, our findings may allow identification of innovative therapy strategies such as the use of modified oligos or small molecules to restore Rbfox2 function in patients with HLHS.To determine regulators that account for abnormal gene expression in HLHS patient RVs, we focused on the RNA binding protein Rbfox2, because it was identified as a major risk allele for HLHS using a large cohort of congenital heart disease patients2021We first checked Rbfox2 mRNA levels in HLHS patient hearts by real time qRT-PCR using gene specific primers . No chanSince Rbfox2 has several spliced variants including a dominant negative (DN) isoform that has a lower molecular weight due to the exclusion of exon 6, which encodes a large portion of the RNA recognition domainSince Rbfox2 HLHS specific nonsense mutation is expected to truncate the protein potentially affecting its nuclear localization, we checked Rbfox2 subcellular distribution in HLHS patient RVs. Neonatal patient hearts with Tetratology of Fallot (TOF) were used as controls for evaluating HLHS specific changes independent of hemodynamic effects on systemic RV dysfunction. Rat newborn and adult, mouse embryonic and human adult hearts were also used as additional controls for determining Rbfox2 localization because of the difficulty in obtaining heart biopsy samples from healthy infants. DAPI/TO-PRO-3 was used to mark the nucleus and cardiac troponin I to mark cardiomyocytes . Using fvs. Ank2 as efficiently as WT Rbfox2 or HLHS specific nonsense mutant of GFP-Rbfox2 in COS M6 cells. WT Rbfox2 was mostly nuclear whereas HLHS specific Rbfox2 nonsense mutant was predominantly cytoplasmic in punctate bodies around the nuclear envelope resembling its pattern seen in HLHS patient hearts vs. 2a. T Rbfox2 . In summBecause Rbfox2 protein was mostly cytoplasmic in HLHS patient RVs, we tested whether transcripts differentially expressed in HLHS RVs are Rbfox2 targets. We used HLHS patient exon array data to extract genomic locations for differentially expressed transcripts in HLHS patient hearts. Using the available HLHS patient exon array data, we extracted sequences for differentially expressed genes in HLHS patient hearts. These sequences were mapped to the human genome to access Rbfox2 CLIP-binding density information10Pnn, Spcs1, Ddx39, Mcm7, Phkb) that were identified as the top changers in HLHS patient RV transcriptome analysisPolr2a)) were shown in Pnn, Spcs1, Ddx39, Mcm7 and Phkb mRNAs were downregulated in HLHS patient RVs when compared to control RVs obtained from Tetralogy of Fallot patients (vs white bar). To determine if Rbfox2 regulates mRNA levels of putative targets altered in HLHS, we ectopically expressed Flag tagged wild type Rbfox2 (Rbfox2WT) or RNA binding deficient mutant (Rbfox2DD). We chose COS M6 cells for this study, as endogenous Rbfox2 levels are very low in COS M6 cells and should not interfere with the overexpressed Rbfox2 proteins. Expression of Flag-Rbfox2WT increased mRNA levels of Pnn, Phkb, and Spcs1 significantly (vs black bars). Expression of the RNA binding deficient mutant of Rbfox2 (Rbfox2DD) did not affect mRNA levels of Pnn, Phkb and Spcs1 demonstrating that Rbfox2 binding ability is necessary to regulate mRNA levels of these putative targets (vs gray bars). Ddx39 mRNA levels were undetectable in these cells. Mcm7 mRNA levels were unaffected in Rbfox2 expressing cells suggesting that change in Mcm7 mRNA levels in HLHS patients are independent of Rbfox2 (data not shown). Rbfox2DD was generated by mutating 2 critical phenylalanine residues in the RNA recognition motif essential for RNA binding activity2627WT and Rbfox2DD mutant proteins were expressed at similar levels in these cells were transfected using FuGENE 6 transfection reagent (Promega E2692) and 1\u2009\u03bcg of Flag or GFP tagged wild type (Rbfox2WT) or RNA binding deficient mutant (Rbfox2DD) or nonsense mutant of Rbfox2 (Rbfox2Nonsense) plasmid DNA according to the manufacturer\u2019s protocol. Corresponding vector DNAs were used as negative controls. Cells were harvested 72\u2009hrs post-transfection for RNA or protein extraction as described previouslyCOS M6 cells were cultured and maintained as described previouslyRNA was extracted from cells and human heart tissues using TRIzol (Invitrogen) according to the manufacturer\u2019s protocol. RNA concentrations were measured using EPOCH Microplate Spectrophotometer (BioTek) and quality of the RNA was assessed using the bioanalyzer at the University of Texas Medical Branch Next Generation Sequencing Core Facility.Ank2 and Roche LightCycler 480 (Roche diagnostic). Specific primers for qPCR were designed to detect the total mRNA levels for Pnn, Spcs1, Ddx39, Mcm7, and Phkb genes with random OligodT primers (Invitrogen) as detailed previouslykb genes . Polr2a 2D gel electrophoresis was performed as described previouslyDD) was generated by mutating two critical Phenylalanine 153 and 155 residues into aspartic acid residues necessary for RNA binding using Quick-change site directed mutagenesis kit (Agilent Technologies Inc.)2627Human Flag-tagged Rbfox2 cDNA clone (transcript variant 6) was generously provided by Dr. Douglas L. Black (UCLA). An Rbfox2 mutant lacking RNA binding activity . Normal human fetal heart protein lysates were purchased from American Life Sciences. Non-diseased fetal heart RNA was purchased from Agilent (pool of 10 hearts from 19\u201321 weeks old fetuses), Biochain (1 heart from 31 week-old fetus) and Clontech (pool of 5 hearts from 25\u201340 week old fetuses). RV biopsy samples from neonates with Tetralogy of Fallot (n\u2009=\u20092 age range 2 to 5 months) were obtained from the Heart Center Tissue Bank at Texas Children\u2019s Hospital.All human heart tissues used in this study were de-identified and pre-existing, and thus were exempt approved by UTMB Institute Review Board (IRB# 11-087). All animal experiments were conducted in accordance with the National Institute of Health Guidelines for the care and the use of animals approved by the Institutional Animal Care and Use Committee of University of Texas Medical Branch (IAUCUC #1101001).How to cite this article: Verma, S. K. et al. Rbfox2 function in RNA metabolism is impaired in hypoplastic left heart syndrome patient hearts. Sci. Rep. 6, 30896; doi: 10.1038/srep30896 (2016)."} +{"text": "Emergency medicine (EM) has undergone rapid development in Rwanda and the delivery of healthcare has steadily improved over the past two decades . Life exCore Content: participants received didactic lectures from US and Rwanda based EM and surgical faculty. The 14 lectures covered basic trauma concepts including shock, head trauma and spinal injuries as shown in Procedure Labs: procedure stations were designed in collaboration with residency directors to address skill deficits of the residents. Didactic sessions were conducted covering proper technique and methodology. Participants were then divided into smaller groups and rotated through two procedure stations: tube thoracostomy and focused assessment sonography for trauma (FAST). Simulation equipment was designed using low cost materials including rehabilitated CPR manniquins and locally available equipment so that participants could replicate this teaching technique with providers at their home institutions.Practicum: on days 1 and 3, participants were randomly assigned to Group A or Group B and underwent pre- and post-training evaluations consisting of a 23-question written exam (Test A or B) and 2 distinct simulation cases (cases 1 and 2 or cases 3 and 4), (See 4), See . All forSimulation Cases: medical simulation cases teaching trauma assessment and management were modified for the Rwandan context. Simulations were performed utilizing course faculty as actors in roles as patient, nurse and case facilitator . Simulation cases received two scores: 1) a completion checklist scored out of 100 and 2) time elapsed to complete pre-identified actions. Themes for the four simulation cases are shown in"} +{"text": "MLH1 hypermethylated tumors with a clear family history.Lynch Syndrome (LS) is caused by pathogenic germline variants in one of the mismatch repair (MMR) genes. However, up to 60% of MMR-deficient colorectal cancer cases are categorized as suspected Lynch Syndrome (sLS) because no pathogenic MMR germline variant can be identified, which leads to difficulties in clinical management. We therefore analyzed the genomic regions of 15 CRC susceptibility genes in leukocyte DNA of 34 unrelated sLS patients and 11 patients with Using targeted next-generation sequencing, we analyzed the entire non-repetitive genomic sequence, including intronic and regulatory sequences, of 15 CRC susceptibility genes. In addition, tumor DNA from 28 sLS patients was analyzed for somatic MMR variants.MLH1 c.1667+1delG). Leukocyte DNA of 11 patients with MLH1 hypermethylated tumors was negative for pathogenic germline variants in the tested CRC susceptibility genes and for germline MLH1 hypermethylation. Somatic DNA analysis of 28 sLS tumors identified eight (29%) cases with two pathogenic somatic variants, one with a VUS predicted to pathogenic and LOH, and nine cases (32%) with one pathogenic somatic variant (n = 8) or one VUS predicted to be pathogenic (n = 1).Of 1979 germline variants found in the leukocyte DNA of 34 sLS patients, one was a pathogenic variant (This is the first study in sLS patients to include the entire genomic sequence of CRC susceptibility genes. An underlying somatic or germline MMR gene defect was identified in ten of 34 sLS patients (29%). In the remaining sLS patients, the underlying genetic defect explaining the MMRdeficiency in their tumors might be found outside the genomic regions harboring the MMR and other known CRC susceptibility genes. MLH1, MSH2, MSH6, PMS2) . As. AsPOLE-patients , 60MLH1 hypermethylated tumors, together with assessment of somatic variants, provides a broad impression of possible genetic causes of tumor formation in MSI-H and/or MMR-deficient tumors. No likely pathogenic MMR gene variants or germline MLH1 hypermethylation were found that explained the familial aggregation of cancer susceptibility in any of the families with MLH1 hypermethylated tumors. With the MMR deficiency of around one-third of the 34 sLS tumors now explained, MMR deficiency in two-thirds of sLS tumors remains genetically unaccounted for. A logical next step is whole exome sequencing (WES) or whole genome sequencing (WGS) to further elucidate the causative genetic defect(s) in the remaining patients.In conclusion, sequencing of the entire genomic region of 15 CRC susceptibility genes in 34 unrelated sLS patients and 11 patients with S1 Table(XLSX)Click here for additional data file."} +{"text": "P < 0.05) with primary important traits . The results of two-way hierarchical clustering demonstrated that Cluster III had outperforming genotype H12 (Sultan \u00d7 Soneri) along with its most closely related traits . Our data shows that H12 (Sultan \u00d7 Soneri) possessed the highest grain yield per plant under environmentally stress conditions, which are most likely to exist in arid and semiarid climatic conditions, such as in Pakistan.This study was carried out to evaluate F1 single cross-maize hybrids in four crop growing seasons (2010\u20132012). Morphological traits and physiological parameters of twelve maize hybrids were evaluated (i) to construct seed yield equation and (ii) to determine grain yield attributing traits of well-performing maize genotype using a previously unexplored method of two-way hierarchical clustering. In seed yield predicting equation photosynthetic rate contributed the highest variation (46%). Principal component analysis data showed that investigated traits contributed up to 90.55% variation in dependent structure. From factor analysis, we found that factor 1 contributed 49.6% variation ( Zea mays L.) is one of the most commonly cultivated crop worldwide .(Suppl. Table S2). Duncan Multiple Range Test (DMRT) for grain yield and its components of four parental lines [see section materials and methods for hybrids and traits description] .(Suppl. Table S3). Duncan Multiple Range Test (DMRT) for grain yield and yield attributing traits of twelve maize hybrids (combined over both seasons) [see section materials and methods for hybrids and traits description].Suppl. Table S4. Eigen vectors (loadings) of first three principal components of yield and yield attributing traits [ see section materials and methods for traits description ]. Suppl. Table S5. Principal factor matrix after rotated factor loadings for yield and yield attributing traits [ see section materials and methods for traits description ]."} +{"text": "Advances in the characterization of allergens have allowed the development of new diagnostic tools based on purified and recombinant allergens. The complex information provided by these multiplexed systems needs a careful interpretation in the light of the local characteristics of patients. Therefore, there is a need to define the sensitisation profiles of each population. The aim of the study was to define specific molecular allergic sensitisation patterns in pediatric polysensitised patients from the Mediterranean area (Barcelona).Pediatric patients experiencing food symptoms who were sensitised to two or more unrelated food groups (excluding milk and egg) by skin prick test (SPT) were included. These patients may or may not also have respiratory symptoms (rhinoconjunctivitis and/or asthma). SPT were performed with standardized food and inhalant allergen extracts. The following parameters were measured: total serum IgE, specific IgE to those allergens shown positive on the SPT, specific IgE to a panel of recombinant allergens by the commercial microarray ImmunoCAP-ISAC\u00ae (Thermofisher Scientific), version 112, that contains 112 individual components. Data analyses were performed using the SPSS Package (release 22.0).120 patients were included with an average age of 11 years (range 4-18). 73% of patients had a family history of allergy. Lipid transfer protein (LTP) was the most prevalent protein sensitisation in our population (74%), followed by storage proteins (57%), tropomyosin (40%) and parvalbumin (29%). Regarding LTP sensitisation, the most frequent molecule determined was Jug r 3 (69%), followed by Pru p 3 (67%) and Pla a 3 (63%), while Jug r 2 (42%), Jug r 1 (38%), Gly m 6 (31%) and Cor a 9 (29%) were the most frequent storage proteins sensitisations shown in our Mediterranean population. There was no statistical association between sensitization to Pru p 3 and anaphylaxis due to vegetable foods (p=0.37). However, as the patient\u2019s age increases, the rate of anaphylaxis caused by vegetable foods rises (p<0.05).Poly-sensitised food-allergic pediatric patients from Barcelona are mostly sensitised to LTP, followed by storage proteins. However, in our population LTP and storage proteins are not associated with increases rates of anaphylaxis. Multiplexed molecular diagnosis delivers added information which may be useful in the management of these patients."} +{"text": "DYT1-DYT25) and sixteen genes have been reported for dystonia. The present study reports the screening of TOR1A (DYT 1) and THAP1 (DYT 6) among primary torsion dystonia patients of India.Dystonia is the third most common movement disorder. It is manifested by involuntary and sustained muscle contractions with frequent twists and repetitive movement, abnormal posture and functional impairment. Genetic factors play significant role for causing dystonia. Till date twenty five loci and 254 controls . All three exons and their flanking sequences including exon-intron boundaries were screened by PCR , sequencing and/or RFLP analysis.First, the most common \u0394GAG mutation and the rs1801968 (p. Asp216His) in TOR1A gene were screened following the published method . THAP1 in 214 patients, a total of five nucleotide variants were identified including a reported missense mutation , a novel heterozygous deletion mutation in two juvenile onset primary dystonia patients and a rare variant in 3\u02b9 UTR region (c. 1030 T>C) in a patient having Blepharospasm. In addition two SNPs, (rs71521601 and rs111989331), were also found both in patients and controls. The association study using rs111989331 (IVS2-87 A>G) demonstrate that the major allele (A) can play as a risk factor for primary dystonia.A total of 321 patients were screened and \u0394GAG mutation was identified in two brothers in a family suffering from primary generalized dystonia. The analysis of rs1801968 (Asp216His) demonstrated that the minor allele (216His) is significantly over represented in patients , suggesting a risk for the disease. On screening TOR1A and THAP1 genes play significant role in the pathogenesis of Indian dystonia patients. This is the first report on mutation detection in TOR1A and THAP1 among Indian dystonia patients.Our preliminary results suggest that the"} +{"text": "Those with single ventricle circulations often have a normal ventricular volumetric ejection fraction (VV EF) which is not in keeping with their symptoms and prognosis. Recently four-dimensional flow (4Dflow) magnetic resonance imaging (MRI) has been used to extract parameters of intra-cardiac kinetic energy (KE). There has been recent interest in measuring ventricular KE in health and disease to see if this could provide an additional tool in the assessment of ventricular function. We propose a new simple measure of ventricular function based on kinetic energy, the kinetic energy ejection fraction (KE EF), and assess its usefulness in patients with single ventricle circulation.Kinetic energy measured by 4D flow MRI was prospectively quantified in 41 patients with a single ventricle circulation (aged 0.5 - 28 years) and compared to 43 healthy volunteers (aged 1.5 - 62 years) who acted as negative controls, and 14 patients with left ventricular (LV) dysfunction (aged 28 - 79 years) who acted as positive controls. The ratio of ejected kinetic energy to total kinetic energy was used to devise KE EF.p = 0.0001). Although VV EF in healthy LVs and single ventricle hearts did not differ (p = 0.99) the respective KE EF was significantly lower with a wider range. Those reporting the greatest symptomatic impairment (NYHA II) had lower KE EF than asymptomatic (NYHA I) subjects (p= 0.0067) whilst in comparison VV EF remained unchanged (p = 0.54).Figure Kinetic energy parameters offer new insight into the function of the heart. KE EF is a marker of healthy cardiac function displaying a very small range of values in comparison to the broader range shown in VV EF. In those with single ventricle circulations it identifies a spectrum of diminished cardiac performance corresponding to a patient's perceived exercise tolerance. This is particularly useful in conditions such as single ventricle physiology where standard ejection fraction values are often normal. Further work to determine its relationship with other prognostic outcomes would allow full implementation as a clinical tool.N/A."} +{"text": "A recent study identified the trithorax group gene Drosophila melanogaster for their role in inducing normal body patterning by positively regulating Homeobox (Hox) gene expression [Hox genes also play critical roles in mammalian development and in specific adult tissues, including the hematopoietic system. Although multiple members of the TrxG gene network interact genetically in flies [Epigenetic modifiers have essential functions in normal hematopoiesis and are frequently dysregulated in hematological malignancies. Trithorax group (TrxG) genes were discovered in pression . Hox genin flies , much leMixed Lineage Leukemia 1 (MLL1) is the mammalian homolog of fly trithorax. MLL1 was originally discovered as a recurrent translocation partner in acute leukemias 3-9, Enhancer-of-zeste and Trithorax (SET) domain with H3K4 histone methyltransferase (HMT) activity that is lost in MLL1 fusion proteins [iewed in ). MLL1 mMLL1, other TrxG members are poorly characterized in mammals. The TrxG gene ash1 was discovered in flies and shown to interact genetically with other TrxG genes [Ash1-like (Ash1l) encodes a large protein that contains a SET domain with in vitro H3K36 histone methyltransferase activity [Ash1l is a critical regulator of adult hematopoietic stem cells (HSCs) [Ash1l-deficient mice generate normal numbers of fetal and neonatal long-term HSCs, but these cells become rapidly depleted in young adult mice. Phenotypically defined Ash1l-deficient HSCs display decreased quiescence and fail to give rise to long-term tri-lineage hematopoietic output after transplantation to irradiated recipients, indicating profoundly defective function. Moreover, Ash1l-deficient HSCs compete poorly for niche space, as evidenced by potent engraftment of wild-type HSCs in Ash1l-deficient recipients even in the absence of irradiation. Despite these defects, Ash1l-deficient mice maintain steady-state hematopoiesis and display enhanced self-renewal of progenitors downstream of HSCs. As observed for Mll1 in normal and malignant hematopoiesis, Ash1l regulates expression of Hoxa cluster genes [Ash1l or Mll1 results in decreased but not abolished Hoxa expression [Mll1 and Ash1l but not either gene alone leads to rapid hematopoietic failure [in vivo demonstration of cooperativity between mammalian TrxG proteins. More work is needed to define the critical pathways operating downstream of Ash1l and Mll1 in HSCs and the molecular mechanisms of their cooperative effects.Besides xG genes . Its mamactivity . We recein vivo therapeutic activity in mouse models of MLL1-driven leukemia, but minimal negative impact on normal hematopoiesis [Men1 and Ash1l in mice results in bone marrow hypocellularity and HSC loss [Men1 loss alone or menin/MLL1 disruption in normal hematopoiesis may result from compensatory activity of the TrxG gene Ash1l. It is also possible that individual molecular pathways differentially regulate recruitment of MLL1 and MLL1 fusion proteins to target genes, opening a therapeutic window. Systematic studies of TrxG gene networks in normal and malignant hematopoiesis could point to the Achilles' heel of MLL1-driven leukemia.As the field moves forward in targeting TrxG proteins in leukemia, careful consideration should be given to understanding how these proteins function during steady-state hematopoiesis in order to exploit critical structural and functional differences. For example, recent work showed that small molecular inhibitors of the menin/MLL1 interaction have substantial opoiesis . Menin iopoiesis and is p"} +{"text": "We examined the concordance of compound-induced transcriptional changes using data from several sources: rat liver and rat primary hepatocytes (RPH) from Drug Matrix (DM) and open TG-GATEs (TG), human primary hepatocytes (HPH) from TG, and mouse liver / HepG2 results from the Gene Expression Omnibus (GEO) repository. Gene expression changes for treatments were normalized to controls and analyzed with three methods: 1) gene level for 9071 high expression genes in rat liver, 2) gene set analysis (GSA) using canonical pathways and gene ontology sets, 3) weighted gene co-expression network analysis (WGCNA). Co-expression networks performed better than genes or GSA when comparing treatment effects within rat liver and rat vs. mouse liver. Genes and modules performed similarly at Connectivity Map-style analyses, where success at identifying similar treatments among a collection of reference profiles is the goal. Comparisons between rat liver and RPH, and those between RPH, HPH and HepG2 cells reveal lower concordance for all methods. We observe that the baseline state of untreated cultured cells relative to untreated rat liver shows striking similarity with toxicant-exposed cells in vivo, indicating that gross systems level perturbation in the underlying networks in culture may contribute to the low concordance.The effect of drugs, disease and other perturbations on mRNA levels are studied using gene expression microarrays or RNA-seq, with the goal of understanding molecular effects arising from the perturbation. Previous comparisons of reproducibility across laboratories have been limited in scale and focused on a single model. The use of model systems, such as cultured primary cells or cancer cell lines, assumes that mechanistic insights derived from the models would have been observed via in vivo vs. in vitro culture systems thought to serve as useful models. The rat liver is by far the most extensively studied model evaluating effects of drugs and other perturbations, and existing data allowed us to assess the level of concordance between rat liver and rat primary hepatocytes cultured in collagen-coated plates (i.e. \u201cflat\u201d culture) for hundreds of drugs. We found that the mouse liver serves as a better model of the rat liver than do rat primary hepatocytes, even after allowing for differences due to pharmacokinetics. The low concordance observed between rat liver and rat hepatocytes suggests that validating the utility of \u2018omics data generated on emerging cell culture approaches using rat cells and comparison to the rat liver may be necessary in order to gain confidence these approaches substantially improve on traditional culture models of human cells.Gene expression studies in model systems are widely used for understanding the mechanism of drugs and other perturbations in biological systems. Other researchers have examined the reproducibility of microarray studies between laboratories, or comparing microarrays and/or RNA sequencing. However, no large scale studies have compared results from protocols which differ in minor details, or results generated Transcriptional changes in model systems are often used to elucidate mechanism-based effects of drug treatment and the relevance for humans , 2. Whilin vivo model using gene expression profiling. Calls to eliminate animal tests in favor of human in vitro models increase the need to understand the relevance of conclusions from gene expression studies across models and species around the solid line (i.e. an arbitrary definition of \u201csimilar performance\u201d). For each axis, increasing self-ID success towards 1 denotes increasing performance.(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.S4 Table(DOCX)Click here for additional data file.S5 Table(DOCX)Click here for additional data file.S6 Table(DOCX)Click here for additional data file.S7 Table(DOCX)Click here for additional data file.S8 Table(DOCX)Click here for additional data file.S9 Table(DOCX)Click here for additional data file.S10 Table(DOCX)Click here for additional data file.S11 Table(DOCX)Click here for additional data file.S12 Table(DOCX)Click here for additional data file.S1 Dataset(XLS)Click here for additional data file.S2 Dataset(XLS)Click here for additional data file.S3 Dataset(XLS)Click here for additional data file.S4 Dataset(XLS)Click here for additional data file.S5 Dataset(XLS)Click here for additional data file.S6 Dataset(XLS)Click here for additional data file.S7 Dataset(XLS)Click here for additional data file.S8 Dataset(XLS)Click here for additional data file."} +{"text": "We and others recently reported the preclinical activity of CIK cells against several solid tumors including mMel [Metastatic Melanoma (mMel) is considered refractory to conventional chemotherapies. New molecular targeted approaches, inhibiting mutated forms of the serine-threonine kinase B-RAF, significantly increased the response rate but patients almost invariably relapse and prognosis remains severe ,2. Open Aim of our research is to explore the preclinical activity of CIK cells against autologous mCSCs surviving treatments with chemo or molecular targeted therapies.We set a preclinical autologous immunotherapy model with primary melanoma cultures and CIK cells generated from patients treated at our Center. To visualize mCSCs we transduced tumor cells with a lentiviral CSC-detector vector encoding enhanced Green Fluorescent Protein (eGFP) under control of the stem-gene oct4 promoter. We treated all mMel cultures with chemotherapy drug fotemustine (IC50 dose). Melanoma cultures harboring BRAF V600E mutations were also treated with BRAF inhibitor (BRAFi) dabrafenib (IC50 dose). We evaluated the presence of residual mCSCs among mMel cells surviving 72h of treatment and explored their susceptibility to immunotherapy with autologous CIK cells.We visualized mCSCs within 11 mMel cultures (3/11 BRAF V600E mutated); median value of eGFP+mCSCs was 15% (range 3.5-26.4%). Putative mCSCs displayed a relative resistance to conventional treatments. The presence of eGFP+mCSCs increased of 39% and 25% among melanoma cells that survived exposure to fotemustine and BRAFi respectively, compared to untreated controls (n=11). CIK cells effectively killed autologous mMel and mCSCs. Tumor specific killing, equally involving bulk mMel targets and mCSCs, ranged between 75% and 33% at effector target ratios of 40:1 and 1:1 respectively (n=8).We provided first proof of concept that putative mCSCs are relatively resistant to conventional treatments with chemo or molecular targeted therapy and susceptible to immunotherapy killing by autologous CIK cells. These preclinical findings support the hypothesis that mCSC may be responsible for disease relapses and support designing of experimental immunotherapy clinical trials with CIK cells in mMel settings."} +{"text": "BRAF V600E mutation (>50% of patients). Patients with BRAF-mutant ECD can respond to BRAF inhibitors. Unfortunately, the lack of adequate archival tissue often precludes BRAF testing. We hypothesized that cell-free DNA (cfDNA) from plasma or urine can offer an alternative source of biologic material for testing. We tested for BRAF V600E mutation in cfDNA from the plasma and urine of 6 ECD patients. In patients with available archival tissue, the result of BRAF mutation analysis was concordant with plasma and urine cfDNA results in all 3 patients . In all 6 patients, BRAF mutation analysis of plasma and urine cfDNA was concordant in 5 of 6 patients . Testing for BRAF V600E mutation in plasma and urine cfDNA should be further investigated as an alternative to archival tissue mutation analysis.Erdheim-Chester disease (ECD) is a rare histiocytosis with a high prevalence of BRAF V600E mutations in at least 50% of patients with ECD.[BRAF mutations can benefit from targeted inhibition of BRAF protein with BRAF inhibitors.[Erdheim-Chester disease (ECD) is a rare form of non-Langerhans cell histiocytosis affecting adults, which is associated with xanthogranulomatous infiltration of foamy macrophages.-3 ECD iswith ECD. FurthermKRAS mutations in urine cfDNA were concordant in 95% of cases with KRAS mutation status in tumor tissue.[BRAF V600E mutation testing in patients with ECD.Cell-free DNA (cfDNA) is released to the circulation from cells undergoing apoptosis, necroptosis and active secretion and has been identified in the plasma and urine of patients with cancer., 8 Detecr tissue. We examiBRAF V600E mutation testing with targeted next-generation sequencing and/or allele-specific PCR in the CLIA-certified laboratory was requested for all 6 patients, but 3 patients had insufficient tissue samples, precluding mutation analysis (Table BRAF V600E mutations were detected in 2 (67%) of 3 tested tumor tissue samples, 3 (50%) of 6 plasma cfDNA samples and 4 (67%) of urine cfDNA samples . Finally, only one patient (patient 1) was treated with a BRAF inhibitor; however, the treatment outcomes were not available at the time of analysis.A total of 6 patients with ECD were enrolled. Their median age at diagnosis was 46 years and most patients were white 4 (67%) and male 4 (67%). Tumor tissue BRAF V600E mutations have been reported in more than 50% of patients with ECD.[BRAF inhibitors such as vemurafenib in ECD patients with BRAF V600E mutations.[BRAF mutations is not feasible in up to 60% of patients.[BRAF inhibitors in general can be effective in patients with BRAF mutations but detrimental in patients without them.[with ECD. In additout them. Thereforout them., 8 cfDNAout them.-14 Arguaout them., 10Our study suggests that mutation analysis of plasma and/or urine cfDNA from patients with ECD can be concordant with archival tissue and should be investigated as its alternative in furthering personalized therapy for patients whose tumor tissue is in short supply.BRAF in tumor tissue were used as a negative control group. Database registration of patients and pathology assessment were performed at MD Anderson. The study and all treatments were conducted in accordance with MD Anderson Institutional Review Board guidelines. A total of 10mL blood samples and approximately 60-120mL of urine from each consented patient were used for DNA isolation.Patients with ECD were referred to the Clinical Center for Targeted Therapy at The University of Texas MD Anderson Cancer Center (MD Anderson) and prospectively enrolled starting in January 2013. In addition, 14 patients with metastatic cancers with known wild-type (wt) Urine cfDNA was isolated by adding urine to an ion-exchange resin . Nucleic acid was eluted with a chaotropic agent and subsequently purified by a silica-based resin . The eluate was further purified using a specific molecular cutoff filter concentrator , followed by a size exclusion column . Plasma cfDNA was isolated using the QIAamp Circulating Nucleic Acid Kit according to the manufacturer's instructions.RNase P, a single-copy gene. Quantified DNA (12.4ng to 60ng) was used for a two-step PCR assay for rare mutant allele detection of a 31bp region containing BRAF V600E and VIC (wt BRAF) TaqMan probes to enable differentiation of mutant versus wild-type quantification, respectively. The RainDrop ddPCR instrument was used for PCR droplet separation, fluorescent reading, and counting droplets containing mutant sequence, wt sequence, or unreacted probe. For a given patient sample, the assay reported BRAF V600E mutation fragments detected as a percentage of detected wt BRAF. Previous to this study, accuracy of the urine-based ddPCR BRAF V600E assay was verified in 89 urine specimens from 50 healthy control samples and 39 samples from 20 patients with known positive BRAF V600E mutation tissue biopsies as determined in a CLIA laboratory.[BRAF metastatic cancer was used to determine a threshold for detection of BRAF V600E mutations. For plasma, >0.094% mutant, equivalent to three standard deviations (0.021%) above the mean of wt BRAF controls (0.031%), was considered positive for BRAF V600E mutation.Urine and plasma cfDNA were quantified by a droplet digital PCR assay to a 44bp amplicon of boratory. Threshol"} +{"text": "Genomic technologies are redefining the understanding of genotype\u2013phenotype relationships and over the past decade, many bioinformatics algorithms have been developed to predict functional consequences of single nucleotide variants. This article presents the data from a comprehensive computational workflow adopted to assess the biomedical impact of the DNA variants resulting from the experimental study \u201cMolecular analysis of sarcomeric and non-sarcomeric genes in patients with hypertrophic cardiomyopathy\u201d \u00a0 Specifications TableValue of the data\u2022These data delineate a prompt informatic pipeline for the prioritization of the most likely pathogenetic DNA variants in a clinical context.\u2022These data are supportive for the researchers to evaluate the prevalence of sarcomeric and non-sarcomeric gene variants in hypertrophic cardiomyopathy.\u2022The described computational strategy is helpful to researchers for the rapid interpretation of Variants of Unknown Significance (VUS) implicated in rare, common and complex diseases.1. DataHere we report the in silico predictions data of the non-synonymous changes found in 41 HCM patients and in 3 HCM-related cases 22.1Nucleotide-specific estimates of evolutionary constraint were explored by (i) GERP++ (Genomic Evolutionary Rate Profiling); (ii) PhyloP placental; (iii) PhyloP veterbrate and (iv) SiPhy.2..2The analysis of intronic variants leading to splicing defects was tested by Human Splicing Finder (HSF) 3.0.2.3The effect of missense changes on the structure and function of a human protein was predicted by: (i) SIFT (Sorting Intolerant From Tolerant), (ii) PolyPhen-2 (Polymorphism Phenotyping v2) HDIV, that identifies human damaging mutations by assuming differences between human proteins and their closely related mammalian homologs as non-damaging; (iii) PolyPhen-2 HVAR, that identifies human disease-causing mutations by assuming common human nsSNPs as non-damaging; (iv) Provean ; (v) LRT (Likelihood Ratio Test) that identifies conserved amino acid positions and deleterious mutations using a comparative genomics data set of multiple vertebrate species; (vi) Mutation Taster; (vii) Mutation Assessor; (viii) FATHMM ; (ix) RadialSVM ; (x) LRT (Logistic Regression Test); (xi) CADD v1.3 (Combined Annotation\u2013Dependent Depletion), a method for objectively integrating many diverse annotations into a single measure (C score) for each variant; and (xii) molecular modeling.indel regions in the structures. Then, ten different models were built for each target protein and evaluated using several criteria. The model displaying the lowest objective function Regarding the molecular modeling, protein structure were experimentally determined by X-ray crystallography, or were inferred by homology modeling means . Protein models were built using the homology modeling approach implemented in modeler-9 package"} +{"text": "Tuberculosis (TB) remains an important cause of death among patients infected with HIV. Genetic variation influences differential development of TB particularly in Immunosupressed patients. Natural Killer cells play an important role in the innate immune system by providing the first line of defence against viral infections and tumors. Their activity is partially controlled by distinct inhibitory and activating killer Ig like receptors (KIR) that recognize specific ligands on potential target cells. Objective of this study is to determine the association between KIR and HIV co-infection.This nested case control study comprised of 68 HIV+ individuals (34 treatment naive HIV-1 slow progressors and 34 on ART patients with TB co-infection). Genomic DNA was extracted from peripheral blood samples by salting out procedure and KIR genotyping was performed for 15 KIR genes (Activating and Inhibitory) by Multiplex PCR sequence specific primer method.All 15 KIR genes were observed in this study cohort. We could observe more number (>4 aKIR) of activating KIR genes in HIV+ TB\u2013 individuals (88%) when compared with HIV+ TB+ patients (74%). Homozygosity for KIR2DL3 was more frequent in HIV+ TB+ patients (64%) than HIV+ TB\u2013 individuals (32%) which may confer susceptibility to co-infection.NK cell receptor KIRs limits HIV disease progression. In the present study, HIV positive individuals who were homozygous for inhibitory KIR2DL3 are at higher risk of acquiring TB co-infection and disease progression with decreased CD4 cells."} +{"text": "We present a fully-crossed factorial mesocosm study and assess how warming and acidification affect the abundance, body size, and fatty acid composition of copepods as a measure of nutritional quality. The experimental set-up allowed us to determine whether the effects of warming and acidification act additively, synergistically, or antagonistically on the abundance, body size, and fatty acid content of copepods, a major group of lower level consumers in marine food webs. Copepodite and nauplii abundance were antagonistically affected by warming and acidification. Higher temperature decreased copepodite and nauplii abundance, while acidification partially compensated for the temperature effect. The abundance of adult copepods was negatively affected by warming. The prosome length of copepods was significantly reduced by warming, and the interaction of warming and CO2 antagonistically affected prosome length. Fatty acid composition was also significantly affected by warming. The content of saturated fatty acids increased, and the ratios of the polyunsaturated essential fatty acids docosahexaenoic- (DHA) and arachidonic acid (ARA) to total fatty acid content increased with higher temperatures. Additionally, here was a significant additive interaction effect of both parameters on arachidonic acid. Our results indicate that in a future ocean scenario, acidification might partially counteract some observed effects of increased temperature on zooplankton, while adding to others. These may be results of a fertilizing effect on phytoplankton as a copepod food source. In summary, copepod populations will be more strongly affected by warming rather than by acidifying oceans, but ocean acidification effects can modify some temperature impacts.Concerns about increasing atmospheric CO The uptake of CO2 by the surface ocean has caused profound changes in marine carbonate chemistry: increased aqueous CO2, bicarbonate (HCO3-), and hydrogen ion (H+) concentrations, while the concentration of carbonate ions (CO32-) declined , which was based on the sum of their individual effects. If the observed combined response of both stressors exceeded their expected additive response then interaction was identified as being synergistic. In contrast, if the observed response was less than the additive response, the interaction was denoted as antagonism [stressorwarm/high \u2013 controlcold/low)] and predicted additive effects, which indicates the direction and magnitude of the interaction.The observed combined impact of both stressors was compared with their expected net additive effect and (B) mean copepod abundance (C1-adult) [ind * L-1]. Error bars denote for \u00b1 1 SD (n = 3). Open symbols represent high pCO2 (1400 \u03bcatm) and closes symbols low pCO2 (560 \u03bcatm) concentrations. Symbols for the treatment combinations as in key.(TIFF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 TableOA (9\u00b0C / 1400 \u03bcatm), OW (ocean warming: 15\u00b0C / 560 \u03bcatm), and OW/OA (15\u00b0C / 1400 \u03bcatm). Values in bold notice significant ANOVA results (see Tables (DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.S4 Tablevs zooplankton biomass. (B) Tukey Honest Significance Test results on phytoplankton vs zooplankton biomass correlation coefficients.(A) ANOVA results of correlation coefficients of phytoplankton Values in bold are significant at p <0.05.(DOCX)Click here for additional data file.S5 Table(DOCX)Click here for additional data file."} +{"text": "RYR1 (ryanodine receptor type 1) are linked to the majority of malignant hyperthermia (MH) families (75%) and some cases of Exertional Heat Stroke. Two CACNA1S variants associated with MH have been functionally characterised. Historically, because of the large size of the RYR1 gene, MH families were screened for 31 diagnostic RYR1 variants only. Methods based on PCR and Sanger sequencing were used. The new opportunity arose since next generation sequencing techniques became available. With this technique sequencing is considerably quicker therefore even long gene sequences such as RYR1 can be easily screened for variations.Mutations in RYR1 and CACNA1S. We applied two different target enrichment methods, Long-Range PCR and HaloPlex (Agilent), to be able to focus on particular regions of the genome.We used Next-Generation Sequencing to look for coding sequence non-synonymous variants of RYR1 and were found negative. Another group consisted of discordant individuals, whose in vitro contracture test (IVCT) phenotype was at odds with their genotype for a familial diagnostic RYR1 mutation. The third studied cohort (n=38) were Exertional Heat Stroke patients, IVCT tested susceptible or normal.In our study we included a group of 80 unrelated MHS individuals who were previously screened for diagnostic mutations in RYR1 variants is not sufficient. Studying the full length of RYR1 and CACNA1S coding sequence may also resolve some phenotype/genotype discordant cases. Finally, Next Generation Sequencing techniques provide opportunities for studying larger gDNA fragments or the whole genome, which may result in discovering some other genes linked to MH.Our results illustrate that Next Generation Sequencing provides broad genetic information quicker and at lower cost than conventional approaches. To improve understanding of malignant hyperthermia genetics, screening of susceptible individuals only for a limited number of diagnostic"} +{"text": "The initiating mechanisms of migraine attacks are very complex but may involve the cyclic adenosine 3',5'-monophosphate (cAMP) signaling pathway. It is unknown whether intracellular cAMP accumulation induces migraine attacks.To investigate whether administration of cilostazol, which causes cAMP accumulation, may induce migraine attacks.We included 14 migraine patients without aura in a double-blinded, placebo-controlled crossover study. All participants received oral cilostazol or placebo on two separate days. We recorded migraine headache characteristics and associated symptoms using a questionnaire.Cilostazol induced delayed migraine-like attacks in 12 patients (out of 14) compared to 2 (out of 14) patients after placebo (P=0.002). The median time to onset for migraine-like attacks was 6 h (range 3-11 h). Patients reported that the attacks mimicked their usual migraine attacks and that cilostazol induced attacks responded to their usual migraine treatment. The median time of medication intake was 6 h (range 4-11).The present study suggests that intracellular cAMP accumulation plays a crucial role in migraine induction. This knowledge is a further step in our understanding of the intracellular pathway of migraine initiation.No conflict of interest."} +{"text": "They were involved in cellular immune response, cell death, and cell survival pathways. Distinct expression signatures based on usage of 308 exons (292 genes) differentiated the groups . This pilot study demonstrates that alternatively spliced genes from whole blood differ in ICH compared to ischemic stroke and differ between different ischemic stroke etiologies. These results require validation in a separate cohort.Whole transcriptome studies have used 3\u2032-biased expression microarrays to study genes regulated in the blood of stroke patients. However, alternatively spliced messenger RNA isoforms have not been investigated for ischemic stroke or intracerebral hemorrhage (ICH) in animals or humans. Alternative splicing is the mechanism whereby different combinations of exons of a single gene produce distinct mRNA and protein isoforms. Here, we used RNA sequencing (RNA-seq) to determine if alternative splicing differs for ICH and cardioembolic, large vessel and lacunar causes of ischemic stroke compared to controls. RNA libraries from 20 whole blood samples were sequenced to 200\u00a0M 2\u2009\u00d7\u2009100\u00a0bp reads using Illumina sequencing-by-synthesis technology. Differential alternative splicing was assessed using one-way analysis of variance (ANOVA), and differential exon usage was calculated. Four hundred twelve genes displayed differential alternative splicing among the groups contains supplementary material, which is available to authorized users. Stroke is diagnosed based on patient history, neurological exam, and brain imaging. Differential diagnosis can be difficult particularly for distinguishing ischemic stroke (IS) from intracerebral hemorrhage (ICH) when imaging is unavailable in the acute setting. Thus, an accurate, inexpensive, and rapid blood test would be useful.Blood transcriptomes show promise as diagnostic biomarkers and have provided insight into understanding the nature of the immune response following human stroke \u20136. HowevAlternative splicing is the process whereby exons from a single gene are included or excluded in the final mRNA transcript or had ICH. IS diagnosis and causes were assessed as described previously , 9. ICH bam files for analysis [Whole blood RNA was used to prepare mRNA libraries using the TruSeq RNA Sample Prep v2 kit and protocol (Illumina). Two hundred million PE 100-bp RNA-seq reads were obtained from each mRNA library using Illumina Solexa sequencing by synthesis on the Illumina HiSeq 2000. TopHat v2.0.7 (Bowtie v2.0.6) was used with default parameters to map reads to a reference genome (Hg19) and generate bam files using featureCounts against AceView (NCBI 37) [p\u2009<\u20090.05), and differential exon usage was assessed between each two groups .The raw reads for genes displaying DAS are shown in Supplementary Table\u00a0NCBI 37) with optNCBI 37) . HoweverNCBI 37) . DAS wasPrincipal components analysis (PCA) and hierarchical clustering were performed in Partek Genomics Suite. Ingenuity Pathway Analysis (IPA\u00ae) and DAVID identified regulated pathways and processes as described previously .Subject demographics and clinical characteristics are presented in Table\u00a0RNA sequencing alignment statistics for all samples among the five groups are presented in Supplementary Table\u00a0p\u2009<\u20090.05; Supplementary Table\u00a0A total of 412 genes displayed differential alternative splicing (DAS) in the whole blood transcriptomes of the five groups of patients with ischemic stroke , intracerebral hemorrhage (ICH) and controls , ICH, and controls is implicated in many human diseases, this is the first study to show that DAS differs between intracerebral hemorrhage (ICH), ischemic stroke, and control subjects. In addition, it is the first study to show that DAS differs between different etiologies of ischemic stroke including cardioembolic, large vessel, and lacunar causes. Identification of DAS in RNA from whole blood for specific stroke etiologies and ICH suggests the immune response varies for each condition. This will be important for understanding the pathogenesis of each condition and will be important for developing biomarkers to differentiate ischemic stroke from ICH and for developing biomarkers to differentiate the different causes of ischemic stroke.This study identified several pathways, molecular functions, and genes previously reported in human ischemic stroke using 3\u2032-biased microarrays , 15. TheThis study is the first to describe genes with DAS and pathways unique for ICH. Among the genes that differentiated ICH from IS were INPP5D (inositol polyphosphate-5-phosphatase) and ITA4 . The INPP5D enzyme regulates myeloid cell proliferation and programming, and its expression correlates with hemorrhagic transformation of ischemic stroke . ITA4 isOther genes with DAS associated with ICH in this study included NAV1 (neuron navigator 1), PDGFC (platelet derived growth factor C), and CCM2 which participate in vascular endothelial growth factor (VEGF) signaling, which predisposes the brain to hemorrhage because of new vessel formation . Of inteSample sizes in this pilot study were small. Thus, we cannot rule out splicing changes due to vascular risk factors. However, hierarchical clustering of the differentially expressed exons demonstrates separation on diagnosis and not on vascular risk factors which are translated into three different proteins which are all derived from a single gene. (GIF 42 kb)High Resolution (TIFF 317 kb)Supplementary Figure 2n\u2009=\u20094), Ischemic Strokes (n\u2009=\u200912) and Control Subjects (n\u2009=\u20094). Dendrograms are not displayed. This is similar to Fig.\u00a0X-axis. Subjects are on the Y axis. Red indicates increased expression and green indicates decreased expression. (GIF 145 kb)Unsupervised Hierarchical Clustering of 308 exons (292 genes) with differential exon usage among Intracerebral Hemorrhage (nar) n\u2009=\u20092 and ConHigh Resolution (TIFF 2305 kb)Supplementary Table 1Over all details of the RNA sequencing reads and quality. (PDF 36 kb)Supplementary Table 2Raw sequencing count for the 412 genes displaying Differential Alternative Splicing (DAS). IS, Ischemic Stroke; CE, Cardioembolic IS; LV, Large Vessel IS; ICH, Intracerebral Hemorrhage. (PDF 60 kb)Supplementary Table 3p\u2009<\u20090.05). (PDF 72 kb)The 412 genes with DAS among Large Vessel Ischemic Stroke (IS), Cardioembolic IS, Lacunar IS and ICH and Controls Canonical pathways of the 412 genes with DAS among Large Vessel Ischemic Stroke (IS), Cardioembolic IS, Lacunar IS, Intracerebral Hemorrhage (ICH) and Control subjects. Benjamini-Hochberg corrected Supplementary Table 5Functions associated with the top 2 molecular and cellular functions over-represented in the 412 differentially alternatively spliced genes. (PDF 113 kb)Supplementary Table 6Raw sequencing count for the 308 exons displaying differential exon usage for Ischemic Stroke , ICH and Controls. (PDF 53 kb)Supplementary Table 7p\u2009<\u20090.0005, FC\u2009>\u2009|1.2|). (PDF 84 kb)Differential exon usage for the 308 exons for Large Vessel IS, Cardioembolic IS, Lacunar IS, ICH and Controls . (PDF 65 kb)"} +{"text": "However, it is not known as to the role of these genes in oxidative stress mediated vaso-occlusive clinical complications of SCD patients.Oxidative stress constitutes one of the significant cause of vaso-occlusive clinical episodes in sickle cell disease (SCD) patients. It brings about the generation of reactive oxygen species and consequent damage to DNA. DNA damage repair genes such as To see the possible association of DNA repair gene polymorphisms with clinical manifestation of SCD patients. Methods: Genotyping of DNA damage repair genes by PCR-RFLP, measurement of oxidant and anti-oxidant status, along with a clinical evaluation of 250 SCD patients and their comparison with normal individuals.hOGG1 gene, XRCC1 gene (codon 280 Arg>His) were found to be significantly higher among SCD patients as compared to controls. However, SCD patients did not show clinical association with any of these DNA repair gene polymorphisms.The level of oxidants were high, and that of antioxidants were low in SCD patients compared to normal individuals. The prevalence of mutant alleles of hOGG1, p53and XRCC1 gene polymorphisms have no clinical association with SCD patients in India.This indicates that Polymorphisms of hOGG1 Ser>Cys, p53 Arg>Pro and XRCC1 have been variably associated with repair of oxidative stress mediated DNA damage in normal human.Sickle cell disease (SCD) patients are usually associated with activation of enzymatic and non-enzymatic sources of reactive oxygen species (ROS) leading to oxidative stress. The resultant oxidative stress leads to microvascular dysfunction, tissue injury and vaso-occlusive crisis led morbidity and mortality among SCD patients.The study sample involved about 250 SCD patients and250 normal individuals (having phenotype Hb AA) from unrelated families belonging to the state of Madhya Pradesh (India). The samples were collected at the sickle cell clinic of Regional Medical Research Centre for Tribals (ICMR), in the the campus of NSB Medical College Jabalpur, Madhya Pradesh (India). The study sample included individuals belonging to Scheduled Caste (SC), Scheduled Tribe (ST) and general population of state of Madhya Pradesh, where both SCD patients and normal individuals were matched with each other with respect the age and ethnicity. Among the study samples, 120 females, 130 males were SCD individuals whereas 118 females and 132 males were normal individuals.The study was initiated after obtaining written consent of patients/their parents and approval by an ethical committee of RMRCT (ICMR), India and NSB Medical College. The patients having recent or multiple blood transfusions and infection and pregnant women were excluded from the study. Besides the present study was conducted in accordance with ethical standards of Helsinki Declaration. Clinical complications such as chest pain, abdominal pain, bone joint pain, history of blood transfusion, the degree of splenomegaly, frequency of blood transfusion were recorded from the date of onset of disease.Oxidant and anti-oxidant status of sickle cell disease and normal individuals were assessed by measurement of plasma/serum level of Vitamin C, Vitamin E, 8-OHdG (8-hydroxy-2\u2032-deoxyguanosine), reduced glutathione (GSH), malondialdehyde (MDA), albumin, activity of lactate dehydrogenase (LDH) and albumin using commercially available Elisa kits and automated biochemical analyzer wherever necessary. Hemoglobin level was measured by an automated blood cell counter .0C 5 min, 35 cycles of 40 s at 950C, 40 s at 600C and 40 s at 720C) was cleaved by Bst UI enzyme (New England Bio Lab) and run in 2% agarose gel.XRCC1 gene, Arg to Gln substitution in exon 10 (codon 399) was amplified to form an undigested fragment of 242 bp using primer pairs 5\u2032 CCC CAA GTA CAG CCA GGT 3\u2032 and 5\u2032 TGT CCC GCT CCT CTC AGT AG3\u2032. The PCR product was digested with Msp1 and analyzed in 2% agarose gel. Homozygous Gln-Gln individuals reflected a single product of 242 bp, homozygous Arg-Arg individuals demonstrated both 148- and 94-bp fragment whereas Arg-Gln individuals revealed all three of the fragments.XRCC1 gene), a 485 bp PCR fragment was obtained using primer pairs 5\u2032 GCC AGG GCC CCT CCT TCA A3\u2032 and 5\u2032TAC CCT CAG ACC CAC GAG T3\u2032. The PCR products were digested with PVU II and analyzed in a 2% agarose gel. Homozygous Arg-Arg individuals reflected a single product fragment of 485 bp whereas homozygous Trp-Trp individuals demonstrated both 396- and 89- bp fragments and heterozygous Arg-Trp individuals revealed all three of fragments.XRCC1 Arg280His codon, a PCR product of 304 bp fragment demonstrates homozygous dominant whereas digestion of PCR product by Rsa I into 246 bp- and 58 bp fragments demonstrate homozygous mutant. Primers used for it were 5\u2032CCC CAG TGG TGC TAA CCT AA3\u2032 and 5\u2032CTA CAT GAG GTG CGT GCT GT3\u2032.XRCC1 gene, PCR conditions were denaturation at 950C 5 min, 35 cycles of 40 s at 950C, 40 s at 600C and 40 s at 720C). Genotyping of hOGG1 Ser 326 Cys was done by PCR-RFLP involving primers \u2013 5\u2032TTG CCT TCG GCC CTG TTC CCC AAG GA3\u2032,5\u2032 TTG CTG GTG GCT CCT GAG CAT GGC CG3\u2032 and restriction enzyme Msp I. The PCR product was 168 bp and MspI digested products were 142, 26 bp for homozygous mutant. The PCR condition was same as described for XRCC1 gene except the annealing temperature kept at 65.50C.12Genotyping of all DNA repair genes gene was performed by PCR - Restriction Fragment Length Polymorphism technique as per published procedure.Fisher\u2019s exact X2 test using the odds ratio (OR), 95% con dence interval were performed by statistical software Graph Pad Prism version 5.0 . The X2 test was also used to test the Hardy-Weinberg equilibrium among the study subjects. The value of P < 0.05 was considered to be significant.Statistical analysis for hOGG1 326Cys (51.8%) and XRCC1 280 His (54.8%) were found to be significantly high in SCD individuals as compared to normal individuals (33.8% and 36.4% respectively). On the other hand there were no significant differences in the frequency of XRCC1 codon 194 and codon 399 alleles as well as mutant allele of p53 genes years, and that of the control group was 17.6 years. The frequency of mutant alleles 53 genes between 53 genes . There w disease .XRCC1 280 His allele and hOGG1 326Cys allele have an increased risk of chromosomal aberrations and many individuals having these mutant alleles have higher incidence of malignancy in normal population.XRCC1 and hOGG1 genes is found to be elevated in a present study of SCD patients; these alleles seem incapable of explaining the usual clinical complications of SCD patients. Rather, adhesion of the sickle cell to endothelium due to oxidative stress cause inflammation that leads to the major cause of clinical symptoms in SCD. 13Clinical complications of sickle cell disease (SCD) involve generation and impairment of oxidative stress. There is a relationship between markers of oxidative stress and common secondary diseases in SCD such as acute chest syndrome and pulmonary hypertension. Autoxidation of sickle hemoglobin (HbS) along with repeated cycle of sickling and unsickling cause premature destruction of erythrocytes and generation of reactive oxygen species (ROS) leading to oxidative stress.XRCC1 280 His allele in normal individuals of the present study is different from the normal population in Asian (7 to 15%), African (3%), Caucasians (5 to 9%). Similarly, frequency of hOGG1 326 Cys allele is different from other countries including earlier Indian study (28 to 29%).Therefore, it appears that there is absence of association between DNA repair gene polymorphisms and clinical symptoms, as reflected in the present study. Frequency of hOGG1, XRCC1, and p53 gene polymorphisms do not seem to play a significant role in clinical manifestations of SCD patients of India.However, further study of SCD populations from different communities of India may give insights on the role of DNA damage repair gene polymorphisms in clinical manifestation of SCD patients. Thus given the present study findings, it may be concluded that hOGG1, p53and XRCC1 gene polymorphisms among SCD patients in India.There is no significant differences in the distribution and clinical impact of"} +{"text": "Stone tool use by wild chimpanzees of West Africa offers a unique opportunity to explore the evolutionary roots of technology during human evolution. However, detailed analyses of chimpanzee stone artifacts are still lacking, thus precluding a comparison with the earliest archaeological record. This paper presents the first systematic study of stone tools used by wild chimpanzees to crack open nuts in Bossou (Guinea-Conakry), and applies pioneering analytical techniques to such artifacts. Automatic morphometric GIS classification enabled to create maps of use wear over the stone tools , which were blind tested with GIS spatial analysis of damage patterns identified visually. Our analysis shows that chimpanzee stone tool use wear can be systematized and specific damage patterns discerned, allowing to discriminate between active and passive pounders in lithic assemblages. In summary, our results demonstrate the heuristic potential of combined suites of GIS techniques for the analysis of battered artifacts, and have enabled creating a referential framework of analysis in which wild chimpanzee battered tools can for the first time be directly compared to the early archaeological record. Pan troglodytes as a model for understanding the origins of technology , , \u201364, but S1 FigDigital Surface Models showing the elevation distribution in the stone tools faces. a) Continuous distribution of the elevation. b) Elevation distribution classified in ten groups defined by the statistical natural breaks of the data according to the Jenk\u00b4s method (ArcGIS 10.2.1).(JPG)Click here for additional data file.S2 FigDigital Surface Models showing the slope distribution in the stone tools faces. Slope measures the rate of change of elevation in the direction of steepest descent. a) Continuous distribution of the slope. b) Slope distribution classified in ten groups defined by the statistical natural breaks of the data according to the Jenk\u00b4s method (ArcGIS 10.2.1).(JPG)Click here for additional data file.S3 FigDigital Surface Models showing the curvature distribution in the stone tools faces. Curvature is the second derivate of the elevation, calculated following the steepest direction (profile curvature), its perpendicular (plan curvature), or a sum of both .(JPG)Click here for additional data file.S4 FigCurvature classification of the stone tools topography based on Dikau 1989, using profile and plan curvatures.(JPG)Click here for additional data file.S5 FigDigital Surface Models showing the roughness distribution in the stone tools faces, estimated through the Terrain Ruggedness Index (TRI), the Vector Ruggedness Measure (VRM), and the 3D/2D area ratio. Roughness models calculated considering the minimum neighborhood (radius = 0.1 mm).(JPG)Click here for additional data file.S6 FigDigital Surface Models showing the roughness distribution in the stone tools faces, estimated through the Terrain Ruggedness Index (TRI), the Vector Ruggedness Measure (VRM), and the 3D/2D area ratio. Roughness models calculated considering a wider local neighborhood (radius = 0.5 mm).(JPG)Click here for additional data file.S7 FigDigital Surface Models showing the value distribution of the Topographic Position Index in the stone tools faces, estimated considering neighborhood radius of 0.1 mm (a), 0.5 mm (b) and 1 mm (c).(JPG)Click here for additional data file.S1 TableElevation (mm), slope (degrees) and roughness (dimensionless) basic statistics of the stone tools, estimated from DSM.(DOC)Click here for additional data file.S2 TablePercentage of curvature classes in the depressions and ridges for every stone tool face.(DOC)Click here for additional data file.S3 Table(DOC)Click here for additional data file.S4 Table(DOC)Click here for additional data file.S1 TextMorphometric analysis and spatial pattern analysis from visual identification.(DOCX)Click here for additional data file."} +{"text": "BRAF, cKIT, CyclinD1) have been evaluated in patients with multiple primary melanoma (MPM).Alterations in key-regulator genes of disease pathogenesis were included into the study. Paired synchronous/asynchronous MPM tissues (N\u2009=\u2009229) were analyzed for BRAF mutations were identified in 109/229 (48%) primary melanomas, whereas cKIT and CyclinD1 amplifications were observed in 10/216 (5%) and 29/214 (14%) tumor tissues, respectively. While frequency rates of BRAF mutations were quite identical across the different MPM lesions, a significant increase of cKIT (p\u2009<\u20090.001) and CyclinD1 (p\u2009=\u20090.002) amplification rates was observed between first and subsequent primary melanomas. Among the 107 patients with paired melanoma samples, 53 (49.5%) presented consistent alteration patterns between first and subsequent primary tumors. About one third of subsequent melanomas presented a discrepant pattern of BRAF mutations as compared to incident primary tumors.The low consistency in somatic mutation patterns among MPM lesions from same patients provides further evidence that melanomagenesis is heterogeneous and different cell types may be involved. This may have implications in clinical practice due to the difficulties in molecularly classifying patients with discrepant primary melanomas. Incidence of cutaneous melanoma has increased during last decades in Western population ,2. Severmitogen-activated protein kinase (MAPK) signal transduction pathway has been reported to play a major role in both the development and progression of melanoma .About one third of patients presented a discrepant pattern of BRAF mutations) among synchronous or asynchronous multiple primary melanomas. Our findings further support evidence that molecular events underlying development and progression of melanoma are really complex. A better comprehension of the factors crucially involved in activating one or the other pathogenetic molecular mechanism, even in the same individual, might have an impact on the disease management. Since the future of melanoma therapy is likely to focus on targeting multiple pathways, advancing technologies will permit to simultaneously investigate multiple genes and targets toward more accurate correlations between molecular signatures and clinical outcome.In our study, we contributed to provide additional clues about the prevalence of alterations in some candidate genes for Click here for filevs. second primary melanomas) for BRAF/cKIT/CyclinD1 alterations.Mutation patterns in patients presenting discrepancies in tumor lesions (7 subsequent Click here for file"} +{"text": "Misdiagnosis of ATTR and late diagnosis may be detrimental hampering adequate management and delaying therapy onset. Objective of the present study was to investigate in a large single-centre cohort of genetically\u2013confirmed ATTR patients the prevalence, type and causes of misdiagnoses. Given the high frequency of cases erroneously diagnosed as having chronic inflammatory demyelinating polyneuropathy (CIDP), we investigated the electrodiagnostic (EDx) features which can help distinguish ATTR from CIDP.Retrospective study design. Review of clinical notes and EDx studies of ATTR patients referred to Amyloid Research and Treatment Centre and C. Mondino National Neurological Institute (Pavia) between 1999 and 2013. EDx of thirty-five patients diagnosed with CIDP were used as control for comparison.Out of 150 patients with ATTR 51(32%) were initially misdiagnosed including 30(59%) CIDP and 11(22%) lumbar spinal stenosis. Eleven (22%) patients underwent spine surgery and 38(74%) were treated with immunotherapies. Patients misdiagnosed had a significant longer delay before diagnosis of 47\u00b13.7 months vs 34\u00b12.7 months (p= 0.01). Lack of family history and onset after 56 years were significantly associated with misdiagnosis(p<0.01). Out of 30 patients misdiagnosed as having CIDP, 17 had original EDx available for review. Six (35%) had definite and 3(17%) possible CIDP according to EFNS criteria, while 8(47%) did not show demyelinating features. Eleven(37%) had a negative tissue biopsy and 4/5(80%) had raised proteins in cerebrospinal fluid (CSF). We next compared EDx of 53 ATTR with EDx from 35 matched CIDP patients. Conduction slowing and prolongation of distal motor latencies were less prominent in ATTR vs CIDP while conduction blocs were almost invariably absent in ATTR. Conversely, in ATTR motor nerves were more often not excitable both at upper and lower limbs.ATTR was misdiagnosed in 1/3 of cases, particularly in patients with late onset and without family history. CIDP was the most common alternative diagnosis, which was supported by EFNS EDx criteria for demyelinating neuropathy in half of them. However conduction slowing is less prominent in ATTR while severe axonal loss is the major EDx feature. DNA testing for TTR should be performed in patients with progressive axonal or mixed axonal-demyelinating peripheral neuropathy, who do not respond to immunotherapies, regardless the lack of family history and a late onset. Raised proteins in CSF and a negative biopsy do not rule out the diagnosis of ATTR."} +{"text": "An integrative study combining phenotypic and transcriptional analysis reveals extensive maternal control of seed size in maize, contributing to a better understanding of the developmental and molecular basis of seed size regulation. 1 crosses were used to investigate developmental and molecular mechanisms governing seed size. Seed morphological characteristics showed striking differences between KLS and KSS inbred lines, and the reciprocal cross experiment revealed a strong maternal influence on both seed weight and seed size. Quantification of endosperm area, starchy endosperm cell size, and kernel dry mass accumulation indicated a positive correlation between seed size, endosperm cell number, and grain filling rate, and patterns of grain filling in reciprocal crosses mirrored that of the maternal parent. Consistent with the maternal contribution to seed weight, transcriptome profiling of reciprocal F1 hybrids showed substantial similarities to the maternal parent. A set of differentially expressed genes between KLS and KSS inbreds were found, which fell into a broad number of functional categories including DNA methylation, nucleosome assembly, and heat stress response. In addition, gene co-expression network analysis of parental inbreds and reciprocal F1 hybrids identified co-expression modules enriched in ovule development and DNA methylation, implicating these two processes in seed size determination. These results expand our understanding of seed size regulation and help to uncover the developmental and molecular basis underlying maternal control of seed size in maize.Seed size is an important component of grain yield and a key determinant trait for crop domestication. The Krug Yellow Dent long-term selection experiment for large and small seed provides a valuable resource to dissect genetic and phenotypic changes affecting seed size within a common genetic background. In this study, inbred lines derived from Krug Large Seed (KLS) and Krug Small Seed (KSS) populations and reciprocal F Zea mays L.) breeding programs, seed size is an important breeding target because of the requirements of both end-use quality and consumer preference is a gene encoding a known endosperm transfer cell-specific transcriptional activator, which is involved in the expression of basal endosperm transfer layer (BETL)-specific genes including BETL-1, Basal layer-type antifungal protein 2 (BAP2), and Maternally expressed gene 1 (Meg1), and plays a central role in the regulatory pathways controlling transfer cell differentiation and associated maternal nutrition allocation the seed coat, which comprises the maternal genotype and imposes mechanical constraints on seed development; (ii) maternal provisioning during seed development; (iii) maternal determination of progeny plasticity in response to developmental signals and environmental cues; and (iv) the effect of the triploid endosperm where gene imprinting occurs most often (miniature 1 (mn1) and shrunken-2 (sh2) were generated from these four parental inbred lines by manual pollination. These hybrids are coded such that the character on the left signifies the maternal parent and the character on the right signifies the paternal parent. Ample pollen was used for each pollination to ensure well-filled ears for consistent kernel phenotyping. Kernels from the center of three different primary ears per plot were either dried for seed weight, fixed in ethanol for imaging, or stored at \u201380 \u00b0C for transcriptional analysis.Thirty cycles of divergent mass selection for seed size were performed in the open pollinated population Krug Yellow Dent to generate KLS30 and KSS30 . Inbred The bulk mature seeds from each plot were used for calculating 100-seed weight measured in grams. To generate the grain filling rate, 10 kernels per ear were sampled at each time point with six replications. Kernel dry weight was determined after drying samples at 65 \u00b0C for 1 week. To obtain the seed size distribution, images were captured using an Epson Perfection V700 Photo desktop scanner and VueScan scanning software without image enhancement and saved as TIFF (tagged image file format) files. One hundred mature dry seeds were spread uniformly with the embryo facing the glass platen and scanned at a resolution of 1200 dpi. Using MatLab image analysis software, the major (depth) and minor (width) axes and total area of the kernels was quantified.http://rsbweb.nih.gov/ij/). Confocal imaging was performed at the Newcomb Imaging Center, Department of Botany, University of Wisconsin-Madison.Kernels freshly isolated from the middle of developing ears at 17 DAP were fixed in 70% ethanol (v/v) and stored in 4 \u00b0C. Kernels were first rinsed with distilled water twice and trimmed on both sides to form a 2\u20133mm thick longitudinal median section containing the embryo. The slices were imaged with a Zeiss AxioZoom fluorescence stereo microscope to obtain the endosperm area. These slices were also stained with 0.1% (w/v) berberine sulfate for 5\u201315min depending on the slice thickness to visualize the cell wall on a Zeiss LSM 510 META confocal microscope. A rectangular region of interest of the approximate location covering most of the endosperm area at 17 DAP was selected and extracted from at least 10 kernels of each of the KLS and KSS inbred lines. Endosperm size and endosperm cell size were measured using ImageJ software , checked for quality, and further purified using an RNeasy MinElute Cleanup kit following the manufacturer\u2019s instructions. Isolation of mRNA, cDNA synthesis, and construction and sequencing of RNA sequencing (RNA-Seq) libraries were performed at the University of Wisconsin Biotechnology Center using the Illumina TruSeq RNA sample preparation kit v2 protocol . Sixteen samples were pooled per lane and sequenced using an Illumina HiSeq 2500 to generate 151 nucleotide paired-end sequence reads. Raw sequence reads are available through the National Center for Biotechnology Information Sequence Read Archive (BioProject PRJNA287557). Quality of the raw sequences was checked using the FastQC program (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) and reads were trimmed to 100 nucleotides to remove low quality bases with the fastx_trimmer program within the FASTX toolkit (http://hannonlab.cshl.edu/fastx_toolkit/index.html). Reads were mapped to the B73 version 2 reference sequence were identified by pairwise comparisons using edgeR \u22640.05 and |log2 fold change| \u22651. A heatmap with dendrograms was produced with the pheatmap R package and there was no overlap in the spread between group L and group S. However, for the paternal groups, the spread overlapped across all four group medians (P>0.05).Significant variation for seed size among the KLS30 and KSS30 populations has previously been shown . Comparisons between group samples that shared either the same KSS maternal parent or the same KLS maternal parent only showed a significant difference in kernel width . L1L1 endosperm had smaller cell area than S1S1 and S3S3 by 32% and 9%, respectively, and this reduction for L3L3 was 26% and 4% in comparison with S1S1 and S3S3, respectively . The relative abundance of transcripts calculated as the FPKM was provided (Supplementary Table S2). We found that 19 909\u201322 443 of 39 456 maize gene models were expressed with FPKM >1 among the samples (Supplementary Table S1). Intriguingly, hierarchical clustering of the transcriptome profiles grouped the samples into four major clades corresponding to the four maternal parents, and reciprocal crosses always formed the primary cluster with their maternal parent . GO enrichment analysis was performed on these four groups of DEGs to discover over-represented functional categories (Supplementary Table S3). No significant GO terms were found enriched in DEGs up-regulated in KSS inbreds at 14 DAP, while up-regulated DEGs in KSS inbreds at 17 DAP were enriched in major metabolic processes such as \u2018carbohydrate metabolic process\u2019 and \u2018\u05b2fatty acid metabolic process\u2019; nutrient reservoir activity was one subcategory of molecular function showing the most marked over-representation in KSS inbreds at both 14 DAP and 17 DAP (see Supplementary Table S3). Significantly over-represented GO terms among DEGs up-regulated in KLS inbreds at 14 DAP were all related to stimulus responses (VIM103) and two DNA methyltransferase genes (MET8 and MET1) , suggestsponse\u2019) . Genes esponse\u2019) . Up-reguSupplementary Table S4). Of the transcription factors up-regulated in KSS inbreds, Prolamin-box binding factor1 (Pbf1) and WRINKLED1 transcription factor 2 (ZmWri1b) from the DOF family and AP2/EREBP gene family, respectively, have been documented as regulators of storage protein and seed oil . The identified modules were then selected on the basis of the module\u2013trait relationship, which was calculated by correlating the module\u2019s eigengene value to the mature seed weight. Thirteen of these modules were found strongly correlated with seed weight , containing between 57 (M13) and 168 (M5) genes . The molecular functions and biological processes that were most significantly enriched in these modules negatively correlated with seed weight were protein autophosphorylation (M1), nutrient reservoir activity (M5), hexose catabolic process (M10), and trehalose biosynthetic process (M13) , Zea mays MADS1 (ZMM1), and ZMM2. According to the maize gene expression atlas . Interestingly, ZAG2 has been identified as a maternally expressed imprinted gene in the maize endosperm in our transcriptome profiling fell into 52 modules, with each containing at least 50 genes (5) genes , and inc5) genes , B. Amonss (M13) . M4 and ss (M13) . Consistss (M13) , which ielopment . This moVariation in seed size is common within and among plant species. Underlying this variation, and thus regulation of seed size, is a complex array of interactions involving genetic factors, developmental signals, and environmental cues. Although maternal effects in plants have long been recognized , the mec1 crosses closely mirrored the phenotype of the self-pollinated maternal parent in terms of seed weight, seed size, and seed development provides strong support for a maternal influence on seed size in maize. The endosperm in cereals serves as the primary nutrient source for embryo and seed development. The endosperm development depends on both sink capacity and assimilates supplied by sporophytic maternal tissues, thus implicating the maternal genotype in the process. The endosperm\u2019s strength as a nutrient sink is proposed to be the function of the number of endosperm cells and/or the number of starch granules formed during grain filling . Therefore, while our study did not focus on identification of imprinted genes, transcriptional differences in genes controlling DNA methylation provide indirect support for the role of gene imprinting as a molecular mechanism underlying the observed maternal effect on seed size.Differential expression and WGCNA analysis both identified DNA methylation as a key process distinguishing large and small seed. We also found a robust association between the DNA methylation GO term and seed size when we compared the current meta-analysis with the previous transcriptional data from ZAG2, its paralogous gene ZMM1, and the C-type MADS gene ZMM2 and SEPALLATA (SEP) in the control of carpel and ovule development in Arabidopsis. ZAG2 was also identified as a maternally expressed imprinted gene in maize by examining the overlap between WGCNA-generated co-expression modules and the imprinted genes identified in developing endosperm genes including ene ZMM2 . AGL genOur comprehensive analyses of seed morphology, endosperm cytology, and seed transcriptome revealed a notable role for the maternal parent in determining seed size. The identification of DEGs and co-expression module genes involved in maternal source constraints extends our understanding of the complex molecular and cellular events in this process and provides a foundation for future studies on seed size in crops.JXB online.Supplementary data are available at Figure S1. Gene clustering tree (dendrogram) for identifying consensus modules obtained by hierarchical clustering of adjacency-based dissimilarity based on FPKM values of all RNA-Seq samples.Figure S2. Heatmaps and barplots of eigengenes for WGCNA-generated co-expression modules that were negatively correlated with seed weight.Figure S3. Spatial and temporal expression of ZAG2 and ZMM1 identified by WGCNA-generated co-expression module M12 based on the Maize B73 Gene Atlas.Table S1. Number of reads, mapping percentage, and number of expressed genes in 24 RNA-seq samples which include four parental inbreds and eight reciprocal F1 hybrids collected at 14 DAP and 17 DAP.Table S2. Gene expression values of B73 RefGen_v2 Filtered Gene Set (FGS) in each sample.Table S3. Differentially expressed genes and Gene Ontology in KLS and KSS inbred lines.Table S4. Transcription factors identified in differentially expressed genes between KLS and KSS inbreds.Table S5. Gene models in co-expression modules significantly associated with seed weight.Table S6. Relationships of the differentially expressed genes and co-expression modules to the maternally expressed gene sets previously described."} +{"text": "The fourth author's name is spelled incorrectly. The correct name is: Xiaoyuan D. Zhang.In the Methods section, there is an error in the last sentence of the first paragrapgh under the heading \u201cAnimal Experiments.\u201d The correct sentence is: (1) Control group (CON): females maintained on a purified control diet ad-libitum throughout pregnancy and lactation; (2) CLA group : females fed a purified control diet supplemented with 1% CLA throughout pregnancy and lactation; (3) High fat group (HF): females fed a purified high fat diet throughout pregnancy and lactation; (4) High fat/CLA group (HFCLA): females fed a HF diet supplemented with 1% CLA throughout pregnancy and lactation."} +{"text": "Capra ibex ibex). At a usually very polymorphic MHC exon involved in pathogen recognition (DRB exon 2), Alpine ibex carried only two alleles. We found that one of these DRB alleles is identical to a DRB allele of domestic goats (Capra aegagrus hircus). We sequenced 2489 bp of the coding and non-coding regions of the DRB gene and found that Alpine ibex homozygous for the goat-type DRB exon 2 allele showed nearly identical sequences (99.8%) to a breed of domestic goats. Using Sanger and RAD sequencing, microsatellite and SNP chip data, we show that the chromosomal region containing the goat-type DRB allele has a signature of recent introgression in Alpine ibex. A region of approximately 750 kb including the DRB locus showed high rates of heterozygosity in individuals carrying one copy of the goat-type DRB allele. These individuals shared SNP alleles both with domestic goats and other Alpine ibex. In a survey of four Alpine ibex populations, we found that the region surrounding the DRB allele shows strong linkage disequilibria, strong sequence clustering and low diversity among haplotypes carrying the goat-type allele. Introgression at the MHC is likely adaptive and introgression critically increased MHC DRB diversity in the genetically impoverished Alpine ibex. Our finding contradicts the long-standing view that genetic variability at the MHC is solely a consequence of ancient trans-species polymorphism. Introgression is likely an underappreciated source of genetic diversity at the MHC and other loci under balancing selection.The major histocompatibility complex (MHC) is a crucial component of the vertebrate immune system and shows extremely high levels of genetic polymorphism. The extraordinary genetic variation is thought to be ancient polymorphisms maintained by balancing selection. However, introgression from related species was recently proposed as an additional mechanism. Here we provide evidence for introgression at the MHC in Alpine ibex , a crucial component of the defense against pathogens, contains the most polymorphic functional genes in vertebrate genomes. The extraordinary genetic variation is generally considered to be ancient. We investigated whether a previously neglected mechanism, introgression from related species, provides an additional source of MHC variation. We show that introgression from domestic goat dramatically increased genetic variation at the MHC of Alpine ibex, a species that had nearly gone extinct during the 18 The MHC is one of the most gene-dense regions and contains the most polymorphic functional genes in vertebrate genomes Balancing selection at a locus may predate speciation events and maintain a set of highly divergent alleles termed ancient trans-species polymorphism Recently, introgression was proposed as an additional mechanism contributing to high levels of genetic diversity at the MHC Capra ibex ibex) is a species of wild goat occupying high-alpine niches of the European Alps spanning from Northern Italy and France to Slovenia. Several related ibex species are found in mountain ranges of Southern Europe, Central Asia, Northeast Africa, and the Arabian Peninsula . One of these species, the bezoar (Capra aegagrus) is the ancestor of the domestic goat th century due to overhunting, Alpine ibex were reintroduced to most parts of the European Alps from the only remaining population in Northern Italy . The reintroduction was very successful, and the species has recovered to more than 40'000 individuals living across the European Alps. Therefore, the Alpine ibex is considered a flagship species of the restoration of large mammals. However, the re-introduction caused several bottlenecks of less than 100 individuals, which substantially depleted genetic variability DRB locus of the MHC class II. Only two alleles (Caib-DRB*1 and Caib-DRB*2) were reported at the exon 2 of DRB in at total of 125 individuals C. aegagrus hircus) and its wild ancestor the bezoar (C. aegagrus) are highly polymorphic at this exon. We show that introgression from domestic goat is responsible for the fact that Alpine ibex are polymorphic at all at the exon 2 of the DRB locus, suggesting that introgression can be an important evolutionary force shaping the evolution of the MHC.The Alpine ibex was identical to the Cahi-DRB*16 allele of the domestic goat first reported from the Japanese breed Shiba (Caib-DRB exon 2 sequence variants (236 bp) as Caib-DRB alleles throughout the manuscript . The frequency of the allele Caib-DRB*2 ranged from 0% to 31% was substantially lower and Chamois (Rupicapra rupicapra) showed even lower mean allele divergence from eight species of the Caprinae subfamily comprising domestic goats and Alpine ibex. With the exception of the sequence shared between domestic goats and Alpine ibex, we found no shared alleles among species. However, we found evidence of shared sequences among species by including sequences of shorter minimum length representing 11 species. Six pairs of species shared 13 alleles . The haplotype of Caib-DRB*1 Alpine ibex individuals did not show any close similarity to sequences found in domestic goats . In contrast, the two haplotypes associated to the Caib-DRB*1 and Caib-DRB*2 allele, respectively, did not show any evidence for recombination . This suggests that Alpine ibex haplotypes associated with Caib-DRB*1 and Caib-DRB*2 alleles, respectively, did not co-exist for a long period in the populations.We tested for the presence of recombinant sequences at the MHC DRB gene, we performed restriction site associated DNA sequencing (RAD-seq). We genotyped 15 Alpine ibex homozygous for Caib-DRB*1 and 15 Alpine ibex carrying Caib-DRB*2 of which 14 were heterozygous for Caib-DRB*2. The sampling covered individuals from four populations. Additionally, we included nine domestic goat individuals (representing four breeds). The RAD sequences were mapped to the domestic goat genome Caib-DRB*2 showed high rates of expected heterozygosity in a region of about 750 kb surrounding the second exon of DRB (24.5 to 25.25 Mb) on chromosome 23 clustered strongly in the section covering the MHC DRB. However, we found no such association of haplotypes containing SNP16397/G at either end of the chromosome and 177 domestic goat individuals (six breeds) using the 52 K Illumina Goat SNP Chip enotyped . We aimeosome 23 . We founromosome . The tigCaib-DRB*2 and two proximal microsatellite markers (OLADRB1 and OLADRB2) covering a physical distance of 107 to 161 kb in the cattle and sheep genome, respectively were observed in the Alpine ibex population with the lowest frequency of the Caib-DRB*2 allele and linkage disequilibria decreased more steeply with pairwise distance between SNPs (2>0.8) were generally shorter (0\u20130.5 Mb) on the chromosomes other than chromosome 23 of Alpine ibex . The non-recombined segment of haplotypes containing an allele under positive selection is expected to be much longer and less diverse than haplotypes containing alleles not under selection, because the latter have experienced recombination or mutation events. The EHH measures the length of such conserved haplotypes on both sides of a specified core SNP Successful introgression of the Caib-DRB*2) were substantially longer and were less diverse than haplotypes carrying SNP16397/A was substantially higher in haplotypes carrying the Caib-DRB*2 than in haplotypes carrying the native Alpine ibex allele, suggesting a substantial increase in allele frequency since the original hybridization event that lead to this introgression. High linkage disequilibria, sequence clustering and increased EHH are consistent with introgression and a selective sweep. We suggest that these signals stem from both the initial introgression event and a subsequent Caib-DRB*2 frequency increase in Alpine ibex populations.The chromosomal region in proximity to the MHC Caib-DRB*2) among all sampled populations across Switzerland and in the Gran Paradiso National Park, the founder population of all extant Alpine ibex populations. The most parsimonious explanation for the introgression of Caib-DRB*2 is that the introgression originated from a successful hybridization between domestic goats and Alpine ibex in the Gran Paradiso National Park prior to the reintroduction of Alpine ibex across the Alps. The domestic goat breed Valdostana bred in the vicinity of the Gran Paradiso National Park shows striking phenotypic similarities to Alpine ibex (http://eng.agraria.org). This is indicative of efforts to interbreed Alpine ibex with domestic goats in this region. We suggest that the Caib-DRB*2 allele was introduced to Swiss populations through animals in the captive breeding program at a period of historically low Alpine ibex population sizes. The low extant frequency of Caib-DRB*2 in the population of Gran Paradiso may be explained by the fact that the population passed through a bottleneck after animals were reintroduced to Switzerland We identified a single introgressed goat allele in a total volume of 50 \u00b5l . For the P1 ligation, adapters containing a unique 6 bp barcode . Size selection (300\u2013700 bp) of the purified DNA fragments was performed using a CALIPER. The excised DNA was purified and blunt-ended (New England Biolabs). 1 \u00b5l dATP and 3 \u00b5l Klenow enzyme (New England Biolabs) were added (30 min at 37\u00b0C) and P2 adapters (1 \u00b5l of 10 mM stock) were ligated . After purification ligation products were PCR amplified using Phusion High-Fidelity DNA polymerase in a total volume of 120 \u00b5l: 50.4 \u00b5l H2O, 60.0 \u00b5l Phusion High Fidelity Master Mix, 2.4 \u00b5l primers (10 \u00b5M), 4.8 \u00b5l library template. Amplification master mixes were divided into 6 separate 20 \u00b5l reactions: 98\u00b0C 30 s; 14 cycles {98\u00b0C 10 s, 65\u00b0C 30 s, 72\u00b0C 30 s}; final extension for 5\u2032 at 72\u00b0C. All purification steps were performed using a MinElute PCR purification kit (Qiagen) according to the manufacturer's recommendations. The library was sequenced on an Illumina HiSeq 2000 platform .RAD library preparation was performed according to http://hannonlab.cshl.edu/fastx_toolkit/index.html) and Trimmomatic 0.30 (www.usadellab.org/cms/index.php?page=trimmomatic) was used for adapter and quality trimming. Reads were then aligned to the domestic goat reference genome The FASTX-toolkit was used for P1 barcode splitting estimates among SNPs were obtained using the R package {genetics}. Heatmaps of pairwise LD estimates were produced using the function LDheatmap in R package {LDheatmap}. In order to investigate the decay of LD over distance, a regression of r2 values against pairwise distance was plotted (2 were computed using non linear least squares (nls in R) as shown in the following function by We constructed haplotype networks of phased SNP data . All LD analyses were performed for each Alpine ibex population and domestic goat breed separately.Where n is the number of haplotypes, The software STRUCTURE {rehh}Measures of the extended haplotype homozygosity EHH, , S6, S7 DRB exon 2 sequences among species of the subfamily Caprinae, a BlastN of Caib-DRB*1 was performed in Geneious version 6.05 . Sequences were aligned using MAFFT v7.023b dist.dna {ape} to calculate genetic distances between all pairs of sequences. As most published sequences were shorter than our sequenced DRB exon 2 fragment, we used a universal 227 bp sequence length to calculate percent identities. We used R to identify sequences shared among species. This comparison was done both for 236 bp (112 sequences unique within species) and 227 bp (332 sequences unique within species).For the comparisons of MHC DRB using the \u03a6w-statistic developed by We tested for evidence of recombination at the MHC Figure S1Genetic clustering of domestic goat and Alpine ibex based on microsatellites. The software STRUCTURE was used to identify the group assignment of 1781 Alpine ibex (yellow) and 182 domestic goats (blue) genotyped at 30 neutral microsatellites [this study and 25]. We also included three known recent F1 hybrids (see zoomed section). Except for the three known F1 hybrids, we did not detect any other recent hybrids (q<0.02). The parameters for the STRUCTURE analysis were as follows: K\u200a=\u200a2, length of burnin period: 20'000; number of MCMC replicates after burnin: 80'000; ancestry model: admixture; allele frequency model: allele frequencies correlated among populations.(EPS)Click here for additional data file.Figure S22), where R2 is the multiple correlation coefficient for a SNP being regressed on all other SNPs in a certain window simultaneously . A VIF of 1 implies that the SNP is completely independent of all other SNPs within the window. We did not detect any recent hybrids (q<0.08). The parameters for the STRUCTURE analysis were as follows: K\u200a=\u200a2, length of burn-in period: 20'000; number of MCMC replicates after burnin: 80'000; ancestry model: admixture; allele frequency model: allele frequencies correlated among populations. STRUCTURE results were consistent when running the analyses either using all 677 SNPs or using different models (admixture model or the linkage model).Genetic clustering of domestic goat and Alpine ibex based on SNP chip data. The software STRUCTURE was used to identify the group assignment of 95 Alpine ibex (yellow) and 177 domestic goats (blue) based on 546 genotyped SNPs. STRUCTURE assumes that markers are either at linkage equilibrium or weakly linked. Therefore, we pruned out loci in strong linkage disequilibrium within species using the option \u201c\u2014indep 50 5 2\u2033 in PLINK threshold: 2). VIF is calculated as VIF\u200a=\u200a1/(1\u2212R(EPS)Click here for additional data file.Figure S3DRB in Alpine ibex carrying Caib-DRB*2. (A) Sliding window of expected heterozygosity for nine domestic goat individuals genotyped using RAD sequencing (window size: 500 kb). (B) Sliding window of expected heterozygosity for 15 Alpine ibex individuals homozygous for Caib-DRB*1 (blue) and 15 Alpine ibex individuals carrying Caib-DRB*2 (green). (C) Number of RAD sequencing SNPs per 100 kb polymorphic in domestic goats (light blue) and Alpine ibex (dark blue).Increased heterozygosity in a 750 kb region surrounding the MHC (EPS)Click here for additional data file.Figure S4Caib-DRB*2 and all chromosomes except the sex chromosome (chromosome 30). The distribution of LD is shown for three different classes: strong (r2>0.8), moderate (0.5A in exon 7) was found in one 4-year-old boy in heterozygous form. The sister of this child had VPA induced fatal hepatotoxicity. The E1143G (c.3428 A>G in exon 21) polymorfism was detected in 7 cases in heterozygous form. We have found in 10 patients the Q1236H (c.3708 G>T in exon 23) substitution in 2 cases homozygous, and in 8 cases heterozygous form.In mitochondrial disorders there is a high prevalence of VPA toxicity predicting genomic alterations. It has a major role in the disease management of these patients. In such cases before VPA treatment genotyping should be recommended to prevent the fatal hepatotoxicity."} +{"text": "The optimum timing of surgery in asymptomatic patients with severe degenerative mitral regurgitation (MR) is controversial and there are no randomised data to separate early repair from watchful waiting. Chronic volume overload may be a stimulus for progressive myocardial fibrosis, which could potentially be an additional marker for left ventricular (LV) recovery after surgery.In total, 24 patients (mean age 62 +/- 16 years) with degenerative moderate or severe asymptomatic MR with no class 1 indication for surgery were recruited from a dedicated valve clinic in a University Hospital and compared with gender matched controls. All subjects underwent cardiac MRI (1.5T Siemens Avanto) for assessment of LV volumes, deformation (SPAMM tagging), late gadolinium enhancement (LE) and T1-mapping using a modified look-locker inversion recovery sequence (MOLLI) before and 15 minutes post gadolinium (0.1 mmol/Kg) for myocardial extracellular volume (ECV). Global ECV was the average measure from basal and mid short axis slices excluding LGE indicative of infarcted myocardium.Patients with MR (mean regurgitant volume 42 ml \u00b1 20) had impaired longitudinal and circumferential systolic deformation Table and incrPatients with MR demonstrate a spectrum of myocardial fibrosis associated with reduced longitudinal and circumferential deformation and reduced peak VO2 max. These changes occur before changes in conventional markers of LV structure and function or an increase in BNP. The routine use of CMR may provide a rationale for improved risk stratification in patients with asymptomatic moderate-severe primary MR to optimise the risk/benefit of early surgery.Nil."} +{"text": "The aim was to evaluate the role of intrathecal lactate as an early predictor of spinal cord injury during thoracoabdominal aortic aneurysmectomy. Forty-four consecutive patients were scheduled to undergo thoracoabdominal aortic aneurysmectomy. Two patients had a type B dissecting aneurysm; all other 42 patients suffered from degenerative aneurysm.During surgery, samples of cerebrospinal fluid and arterial blood were simultaneously withdrawn to evaluate lactate concentration. Samples were collected at five fixed times during and after surgery: T1 (beginning of the intervention), T2 (15 minutes after aortic cross-clamping), T3 (just before unclamping), T4 (end of surgery), and T5 (4 hours after the end of surgery).(P = 0.04) higher than those of the other 40 patients preoperatively (2.12 \u00b1 0.35 vs. 1.79 \u00b1 0.29 mmol/l).Mean lactate levels in cerebrospinal fluid rose consistently from the beginning of the intervention steadily until after surgery . Seven patients developed spinal cord injury; two of them had delayed injury occurring 24 hours after the end of surgery; the remaining five had early onset. In this group of five patients, preoperative cerebrospinal fluid lactate levels were significantly The preoperative cerebrospinal lactate concentration is elevated in patients who will develop early-onset spinal cord injury after thoracoabdominal aortic aneurysmectomy. This may allow a better stratification of these patients, suggesting a more aggressive strategy of spinal cord function preservation and possibly guaranteeing them a better outcome."} +{"text": "These AA-induced changes contribute to the limitation/amelioration of colitis pathology. Investigating strategies to manipulate and modulate different aspects of these pathways would offer novel insight into inflammatory bowel disease pathogenesis, and will augment the development of new therapeutic options to manage recalcitrant colitis.The appendix contains copious lymphoid tissue and is constantly exposed to gut flora. Appendicitis and appendectomy (AA) has been shown to prevent or significantly ameliorate ulcerative colitis. In our novel murine AA model, the only existing experimental model of AA, the appendiceal pathology closely resembles that of human appendicitis; and AA offers an age-, bacteria- and antigen-dependent protection against colitis. Appendicitis and appendectomy performed The vermiform appendix or simply the appendix, contains copious lymphoid tissue and is constantly exposed to gastrointestinal bacteria. Appendicitis is the most common abdominal emergency requiring surgery , its hig1) In mice, the major caecal lymphoid patch is the equivalent of the human appendix Balb/c mice , are intraperitonealy anaesthetised with xylazine and ketamine followed by allocation into 2 treatment groups, the appendicitis group or the sham surgery group .\u2022\u2003Other permutations (appendicitis only or appendectomy only) do not ameliorate colitis pathology, and hence are excluded.\u2022\u2003Mice are randomised to have either appendicitis or sham operation.(3) Appendicitis is induced by constructing an appendiceal pouch from the caecal lymphoid patch, which is the equivalent of the human appendix.\u2022\u2022This appendiceal pouch Figure\u00a0 is obstr(4) Sham surgery entails a similar procedure, but without continuous obstruction by band ligation of the caecal patch.\u2022\u2003It also involves the placement of a sterile rubber band in the abdominal cavity as a control for foreign body reaction.\u2022\u2003While most human ulcerative colitis patients would not have undergone any form of abdominal surgery at diagnosis, it is essential that we control for the colonic and peritoneal milieu changes induced by a simple laparotomy in our experimental AA mice group.\u2022\u2003Additionally, the small rubber ring used to encircle the main caecal follicle in the AA group needs to be controlled for too. Hence, in each SS group mouse, a rubber ring is introduced in the peritoneal cavity via a small laparotomy incision.(5) Seven days following initial surgery, appendicitis mice undergo appendectomy by ligation and excision, while sham mice undergo a second sham surgery .(6) All mice are monitored daily.(7) Distal colons are harvested (3 days or 14 days or 28 days post-AA) without the recta - a 1 cm-segment (from the anus) are excluded prior to harvesting the distal colon.(8) Transmural distal colonic segments are chosen to minimise artifactual changes and maximise pathophysiological relevance; as not only superficial mucosal tissue, but also colonic lymphoid nodules/aggregations and other tissue may \u201ctransmit\u201d the protective immunological changes induced by appendicitis and appendectomy to the distal colon.\u2022\u2003The proximal colon would be much more likely to be affected by the inflammatory changes experimentally induced by AA, owing to the close anatomical proximity of the main mouse caecal patch (\u2248human appendix) to the proximal colon.\u2022\u2003The novelty of our study is that the most distal regions of the large gut sustain major persistent immunological changes (protective against colitis), by manipulation at the caecum, the most proximal region of the large gut., 15596-026), snap-freezing in liquid nitrogen, and storing at -80\u00b0C until the microarray analysis.(9) The processing of transmural distal colonic samples consists of flushing out of faecal contents with normal saline and immediately transfer to TRIzol\u00ae reagent After chloroform and isopropanol treatment and centrifugation followed by washing the resultant pellet with 75% ethanol, air-drying and final re-constitution in nuclease-free H(11) Concentration and purity of RNA is determined by automated optical density evaluation (OD 260/OD 280\u2009\u2265\u20091.8 and OD 260/OD 230\u2009\u2265\u20091.8) using Nanodrop ND-1000 .(12) The degree of RNA degradation is analyzed by the Agilent electrophoresis bioanalyzer 2100 with the RNA integrity number (RIN) values consistently above 7.(1) Fine forceps and scissors(2) Autoclaved bands (rings) and transfer pipettes(3) Suture holder and sutures (Needle Eithlon 4-0)(4) Syringes and needles (22G)(5) Surgical platform and sterile gloves(6) Sterilised distilled water, Falcon tubes, and tube holder frame(7) P1000 pipettes and corresponding sterilized tips (with prior tailor-made bevelling)(8) Sterile dressing packs (9) Shaving apparatus with shaving cream(1) 50 ml of sterile Phosphate Buffered Saline (PBS)(2) Sterile saline (Sodium Chloride Injections BP 0.9% tubes)(3) 5 ml anaesthetic stock solution composed of Ketamine/xlyazine/sterile PBS - A standard 5 ml anaesthetic stock solution would contain Ketamine (100mg/ml) 1 ml, Xylazine (20mg/ml) 0.25 ml, and Sterile PBS 3.75 ml.Ketamine (100 mg/ml) - 1ml for 100 mg/kg\u2022\u2003Xylazine (20 mg/ml) - 0.17 ml for 3.4 mg/kg OR 0.25 ml for 5 mg/kg\u2022\u2003Sterile PBS - 3.83 ml for 3.4mg/kg Xylazine OR 3.75 ml for 5 mg/kg Xylazine\u2022\u2003(4) Spray bottle with 70% Ethanol(5) Povidone-iodine(1) Shave a circumscribed area of each mouse\u2019s abdomen (left side) a day before surgery.(2) Weigh each mouse and make identification marks on each mouse\u2019s tail a day before surgery.(3) Wrap each P1000 pipette with sterile plastic sheet (from dressing pack).(4) Anaesthetise mouse with ketamine/xylazine/sterile PBS \u2013 Intraperitoneal injection of 0.1 ml of anaesthetic stock solution per 20 g mouse weight.(5) Prepare surgical area observing sterile conditions meticulously:\u2022\u2003Use sterile gloves throughout.\u2022\u2003Cover bench with sterile plastic sheet and tape the corners on the hood bench.\u2022\u2003Use sterile plastic forceps to pick up the apparatus from the pack and put aside.\u2022\u2003Load autoclaved equipment onto sterile plastic sheet by the \u201cno touching\u201d method.\u2022\u2003Load bands, and pour Povidone-iodine onto sterile tray.(6) Load band onto bevelled pipette tip with fine forceps and set aside.(7) Clean surgical platform with ethanol and place on bench.(8) Fix mouse to the platform by taping their arms/legs with taps.(9) Moist eyes with a drop of saline to prevent xerosis and keratitis.(10) Wipe abdomen with ethanol and clean the left part of the shaved abdominal wall with Povidone-iodine.(11) Cover mouse with the sterile draper from dressing pack and make a hole in the draper above surgical area with fine forceps.(12) Perform a ~1cm laparotomy on the left sided abdomen, using fine forceps to pick up skin and peritoneum.(13) Mobilise caecum out to the surface with blunt curved forceps.(14) Frequent moistening of the caecum with PBS is essential.(15) Identify the 2 to 4 white appendiceal lymphoid patches, with one being far bigger than the others \u2013 this would be the one to do the procedure on.(16) Attach bevelled pipette tip with band to pipette, and aspirate the lymphoid patch with firm suction pressure in gentle, but firm circular motion. Maintain suction pressure for 10 seconds before slowly unloading the band (with fine forceps) onto the neck of the newly created pouch.\u2022\u2003Include some colonic tissue during suction because the created pouch will get smaller as each mouse ages.(17) Using a curved forceps carefully return bowel into abdominal cavity by gently grasping the band.(18) Dispense 1ml of PBS in the abdominal cavity to moisten the peritoneum.(19) Suture the abdominal wall and the skin in layers.(20) Give 1 ml normal saline subcutaneously (scruff of neck) after the procedure.(21) Place mice on their side and put them close to each other in the cage to prevent hypothermia.(22) Check post-op recovery in 1 hour for complete recovery. Check recovery for 1 week post operation and record observations in post operative period table.(23) Appendectomy is performed by 7 days after appendicitis induction by gentle separation of adhesions, ligation, and excision of the inflamed area distal to the ligature.(24) The mice are sacrificed 3 days or 14 days or 28 days after that for distal colon harvesting.Distal colonic gene expression studies; 3 days after surgery, or 28 days after surgery; reveal the various genes and gene-sets that are responsible for the durable protective effect of AA against colitis . The genin the most proximal colon induces the following gene expression changes in the most distal colon Substantial curbing of T helper 17 cell -recruitment, -differentiation, -activation, and \u2013effector (interleukin) expression contributing to suppression of Th17 pathway-mediated immunopathologial damage in colit(2) Significantly and globally curbing autophagy gene expression , contribLate but significant suppression of genes and gene-sets pertaining to endothelin activity - This would suppress endothelin vasoactivity-mediated immunopathologial damage and vascular remodelling in inflammatory colitis (Manuscript submitted).(3) (4) No changes in interferons; but upregulation of genes pertaining to specific IFN activity-associated molecules, and downregulation of genes pertaining to other IFN activity-associated molecules (Manuscript submitted).Using our unique murine AA model, elucidating the pathways involved in these changes will enhance the development of approaches and techniques to manipulate different genes, enzymes, and proteins related to these pathways; towards improving therapeutic options in IBD. Investigating strategies, involving monoclonal antibodies, combinatorial peptides, and small molecules (identified by high throughput screening) to manipulate and modulate different aspects of these pathways, would augment the development of new therapeutic options to manage IBD.AA: Appendicitis-appendectomy; SS: Sham-sham; IBD: Inflammatory bowel disease; TNBS: Trinitrobenzene sulfonic acid.The author declares that he has no competing interests."} +{"text": "Sonic hedgehog (Shh) signaling pathway during mouse embryonic development, but its exact role and mode of mechanism have remained elusive. Since modulation of Shh signaling depends on normal functioning of primary cilia, and Evi5L's overexpression (Rab23's RabGAP) led to decreases in primary ciliogenesis, Rab23 likely has a role at the cilium. We found Rab23 wild-type and constitutively active Rab23Q68L mutant enriched at the primary cilia. In testing the effect of Rab23's manipulations on the ciliary localization of several known ciliary cargoes, ciliary localization of a kinesin-II motor protein Kif17 turned out to be disrupted by overexpression of dominant negative Rab23S23N and in Rab23 depleted cells. In addition, Kif17's ciliary mislocalization could be rescued after expression of a non-degradable wild-type Rab23 and FRAP experiments showed that Rab23 silenced cells exhibited reduced recovery of ciliary Kif17 after photobleaching. Co-immunoprecipitation studies further revealed that Rab23 exists in a tripartite complex with Kif17 and Importin \u03b22 (Kif17's ciliary import carrier), implying that Kif17 requires binding to regulatory proteins like Rab23 for its ciliary transport. Although a ciliary-cytoplasmic gradient of nuclear Ran is necessary in regulating Kif17's ciliary import, both small GTPases Rab23 and Ran appear to have independent roles in the ciliary entry of Kif17. Our findings have uncovered a hitherto unknown effector of Rab23 and demonstrated how Rab23 could mediate Kif17's trafficking to the primary cilium.Rab GTPases are regulators of the intracellular vesicular transport and interorganelle protein trafficking of both endocytic and secretory pathways. The small GTPase Rab23 is an antagonist of the"} +{"text": "Investigators from Danish Headache Centre, Glostrup Hospital, and the University of Copenhagen investigated whether intracellular cyclic AMP accumulation induced migraine attacks among 14 migraine patients without aura. P = 0.003). The headaches induced were similar to their usual migraines and were effectively aborted by their usual migraine treatment. Nausea was significantly different between the two groups (P = 0.027). Photo- and phonophobia were not significantly different between groups. The authors suggest that the intracellular accumulation of cyclic AMP induces migraine attacks, which furthers our understanding of the migraine pathophysiology. [Investigators from Danish Headache Centre, Glostrup Hospital, and the University of Copenhagen investigated whether intracellular cyclic AMP accumulation induced migraine attacks among 14 migraine patients without aura. Patients either received placebo or cilostazol, a selective inhibitor of phosphodeisterase type 3 (PDE3) which prohibits the breakdown of cyclic AMP (cAMP). Patients were instructed to record their headache symptoms, localization and characteristics by a self-administered questionnaire every hour until 13h post-administration. Twelve of 14 patients (86%) developed migraine-like attacks after cilostazol compared with two patients (14%) after placebo. The median time to migraine onset was 6h post cilostazol (range 3-11). Eleven of 14 (79%) took rescue medication cilostazol and 2 (14%) after placebo has been well established to cause migraines. Studies in healthy volunteers and migraine patients with sildenafil, a highly selective inhibitor of phosphodiesterase type 5 (PDE5) showed that cyclic guanosine monophosphate (cGMP) and cAMP are likely mediator of headache responses elicited by NO . Calcito"} +{"text": "The rapid development and refinement of techniques over the past decade have led to the realization that a Minimally invasive (MI) approach enables complex (open) valve surgery to be performed with results equivalent to those of conventional valve surgery done in experienced center. Rheumatic Heart Disease (RHD) continues to be endemic in developing countries like India.To compare minimally invasive surgery with standard sternotomy procedure among patients of rheumatic mitral valve disease.A retrospective comparative study was carried out in tertiary care hospital of central India. From April 2013 to December 2014, a total of 128 eligible study subjects [range 7 to 66 years] presented to CVTS department of our hospital were included. Majority of were in NYHA III (52.9%) and NYHA IV (29.7%). All study subjects were considered for minimally invasive (MI) approach, however only 63 study subjects fulfilled the eligibility criteria for the above procedure. Clamp time (CT), Cardiopulmonary bypass (CPB) time, intensive care unit (ICU) stay and hospital stay was observed among study subjects. These results were compared to 63 age and sex matched study subjects from 2012 and also with 65 study subjects who had not undergone minimally invasive surgery from the same period.Among MI surgery, operative mortality was 02 (3%), re-exploration were 3 (5%) and 3 (5%) re-admission. 01 re-admission was for delayed tamponade and 02 were for non-compliance with oral anticoagulants. Statistically significant lower CT, CPB time and ICU stay was found in minimally invasive surgery when compared with standard sternotomy approach (p < 0.006).Minimally invasive approach for rheumatic valve yields similar morbidity and mortality with reduced surgical time and length of ICU stay. Peripheral cannulation does not cause problem and cosmetic results were excellent."} +{"text": "Felis catus gammaherpesvirus 1 (FcaGHV1) from lymph node DNA of an infected cat. The genome includes a 121,556-nucleotide unique region with 87 predicted open reading frames (61 gammaherpesvirus conserved and 26 unique) flanked by multiple copies of a 966-nucleotide terminal repeat.We sequenced the complete genome of Felis catus gammaherpesvirus 1 (FcaGHV1) is a newly identified virus which clusters phylogenetically with members of genus Percavirus, subfamily Gammaherpesvirinae, family Herpesviridae (ing MIRA as well ing MIRA . Contigsing MIRA and PRICing MIRA to produing MIRA .We defined open reading frames (ORFs) by prediction with GeneMarkS and FGENKT595939.The FcaGHV1 genome sequence was deposited in GenBank under accession no."} +{"text": "Bronchopulmonary dysplasia (BPD), a chronic lung disease of preterm infants, is characterized by impaired alveolar growth and pathologic vascularization.Fgf10 expression deficiency on embryonic lung development and hyperoxia lung injury (BPD mouse model).To investigate the impact of Fgf10+/- and Fgf10+/+[WT]) were harvested at E12.5 and E18.5. Lung branches were quantified and H&E staining was performed.Embryonic lungs , gene expression analysis , FACS were performed at P3.Fgf10 heterozygous lungs exhibit epithelial branching defects and decreased Fibroblast growth factor signaling.Embryonic Fgf10+/- lungs show decreased epithelial proliferation.Embryonic Fgf10+/- embryos display structural defects and abnormal gene expression.Lungs of E18.5 Fgf10 heterozygous pups display increased sensitivity to hyperoxia exposure associated with significant structural lung defects. Transcriptomic analyses show epithelial defects linked to cell cycle dysregulation and increased Tgfb signaling. Fgf10 heterozygous vessels are more sensitive to hyperoxia injury and exhibit a less muscularized phenotype. SPC staining shows significant decrease in SPC+ cells after hyperoxia injury. FACS analysis revealed an increase of AEC I on the expenses of AEC II (progenitor cell).Fgf10 deficiency leads to impaired embryonic lung development and death upon postnatal hyperoxia lung injury due to vulnerability of the epithelium."} +{"text": "Out-of-Hospital Cardiac Arrest (OOHCA) is associated with a poor prognosis. Targeted temperature management (TTM) within Intensive Care (ICU) including therapeutic hypothermia (TH) aims to reduce cerebral reperfusion injury and improve neurological outcomes.\u00a9, Alsius UK\u00ae)Within Northern Ireland (NI), Craigavon Area Hospital (CAH) is the only ICU to implement TH using an intravascular cooling device , were identified from the Intensive Care National Audit and Research Centre (ICNARC) database.35 patients (87.5%) had available relevant and complete data.Overall our outcomes for a mixed ICU population with broad inclusion criteria for TH are comparable with those of published studies .The use of intravascular cooling for TH was associated with minimal use of muscle relaxants allowing early neurological prognostication in our patient group.However intravascular cooling to 32-34\u00b0C was associated with prolonged periods of rebound hyperthermia in a significant minority of patients .We believe that TH to 32-34\u00b0C, using intravascular cooling, increases the risk of developing a rebound hyperthermia that could potentially exacerbate acquired neurological injury.Our data supports the use of TTM to 36\u00b0C to mitigate any potential effect of rebound hyperthermia is this patient group."} +{"text": "This article was republished on February 23, 2016 to remove or replace copyrighted material and images with privacy issues in two Supporting Information files, S1 PowerPoint(PPT)Click here for additional data file.S2 PowerPoint(PPT)Click here for additional data file."} +{"text": "Vernicia fordii) provides the sole source of tung oil widely used in industry. Lack of fatty acid composition and molecular markers hinders biochemical, genetic and breeding research. The objectives of this study were to determine fatty acid profiles and develop unigene-derived simple sequence repeat (SSR) markers in tung tree. Fatty acid profiles of 41 accessions showed that the ratio of \u03b1-eleostearic acid was increasing continuously with a parallel trend to the amount of tung oil accumulation while the ratios of other fatty acids were decreasing in different stages of the seeds and that \u03b1-eleostearic acid (18\u22363) consisted of 77% of the total fatty acids in tung oil. Transcriptome sequencing identified 81,805 unigenes from tung cDNA library constructed using seed mRNA and discovered 6,366 SSRs in 5,404 unigenes. The di- and tri-nucleotide microsatellites accounted for 92% of the SSRs with AG/CT and AAG/CTT being the most abundant SSR motifs. Fifteen polymorphic genic-SSR markers were developed from 98 unigene loci tested in 41 cultivated tung accessions by agarose gel and capillary electrophoresis. Genbank database search identified 10 of them putatively coding for functional proteins. Quantitative PCR demonstrated that all 15 polymorphic SSR-associated unigenes were expressed in tung seeds and some of them were highly correlated with oil composition in the seeds. Dendrogram revealed that most of the 41 accessions were clustered according to the geographic region. These new polymorphic genic-SSR markers will facilitate future studies on genetic diversity, molecular fingerprinting, comparative genomics and genetic mapping in tung tree. The lipid profiles in the seeds of 41 tung accessions will be valuable for biochemical and breeding studies.Tung tree ( Vernicia fordii) is a native woody oil plant in subtropical areas of China. This important economical tree has been grown in China for the production of tung oil or ornamental garden for centuries cis, 11trans, 13trans octadecatrienoic acid) Tung tree or tung oil tree V. fordii clones were released in China in the 1980s for cultivation on the basis of field survey, collection and evaluation data V. fordii germplasm resources has been more widely recognized. We initiated germplasm collection in 2007. Some superior germplasm were collected from main distribution areas of V. fordii in China and planted at the Central South University of Forestry and Technology Germplasm Repository.Understanding fatty acid composition and genetic diversity among tung tree germplasm resources is essential for tung tree breeding and clonal improvement. A series of elite V. fordii-specific EST-SSR markers V. fordii genomic DNA by AFLP of Sequences Containing repeats protocol Microsatellites, also known as simple sequence repeats (SSRs) or short tandem repeats, are repeating sequences of 2\u20136 base pairs of DNA Great progress has been developed in high throughput sequencing technology, i.e. Next Generation Sequencing, utilizing the Roche/454 Genome Sequencer FLX Instrument, the ABI SOLiD System and the Illumina Genome Analyzer. These new sequencing technologies not only offer fast, cost-effective and reliable approaches for the generation of large expression-data sets in both model and non-model plants with large and complex genomes The objectives of this study were to evaluate fatty acid profiles and develop unigene-derived SSR markers in 41 tung tree accessions collected from five Provinces in China. Gas-chromatography (GC) analyzed fatty acid profiles in the mature and developing seeds. We utilized Illumina platform-based transcriptome sequencing of cDNA library from developing tung seeds and characterized microsatellites from the transcriptome sequences and developed 15 new polymorphic genic-SSR markers. We also analyzed the expression levels of the identified polymorphic SSR-associated unigenes in developing tung tree seeds and correlated their expression levels with oil content and fatty acid composition in the seeds. The fatty acid composition profiles and novel genic-SSR markers will be useful for biochemical and genetic research and tung tree improvement.Vernicia fordii, a diploid plant) were collected from Henan (HEN), Hunan (HUN), Hubei (HB), Guizhou (GZ) and Shanxi (SX) Provinces in China. Collecting the samples did not require specific permits because the trees were public-owned and the field studies did not involve protected species. These tung trees were planted at Central South University of Forestry and Technology Germplasm Repository. Vouchers of the sampled accessions were deposited in the University\u2019s Herbarium. Forty-one cultivated accessions at 4-year old were used in this study. The voucher numbers, original locations and geographical coordinates of these 41 tung tree accessions are described in Tung trees .Genomic DNA was isolated from young leaves of the 41 Tung seeds from accession HUN42 were selected because its seeds contained the highest amount of seed oils among the 41 accessions and exhibited a typical lipid profile. The seeds were collected at lipid synthesis initiation phase , peak phase magnetic beads and fragmented into 200\u2013500 bp pieces using divalent cations at 94\u00b0C for 5 min. The cleaved mRNA fragments were reverse transcribed into first-strand cDNA using SuperScript II reverse transcriptase and random primers (Life Technologies). After double-stranded cDNA synthesis, fragments were end repaired and A-tailed. The final cDNA library was created by purifying and enriching the above products with polymerase chain reaction (PCR).The cDNA sequences were determined through a paired-end flow cell using an Illumina Solexa HiSeq 2000 Sequencing System at Beijing Genomics Institute . The clean reads after DNA sequencing were de novo assembled using Trinity with default K-mers\u200a=\u200a25 The microsatellites were detected from the assembled unigenes using the MIcroSAtellite tool 2, 200 \u00b5M of each dNTPs, 0.2 \u00b5M of each primer and 0.25 unit of Taq DNA polymerase . The PCR conditions were set at 95\u00b0C for 5 min, 35 cycles of 30 s at 94\u00b0C, 30 s at 56\u00b0C and 30 s at 72\u00b0C and a final extension of 10 min at 72\u00b0C on a DNA Engine thermal cycler . The amplification products were resolved on 2% agarose gels.Genomic DNAs from leaves of three tung tree accessions were used for testing PCR primers corresponding to 98 loci by agarose gel electrophoresis. PCR primer pairs were designed using Primer Premier 5.0. The parameters for primer design were set for primer length from 18 to 26 nucleotides, PCR product size from 100 to 400 bp and annealing temperature from 50\u00b0C to 60\u00b0C. The sequences of the forward and reverse primer pairs for 98 unigenes tested, the SSR repeated motifs and the amplicon sizes of PCR products are described (5\u2032-TGTAAAACGACGGCCAGT-3\u2032) sequence-tag method The loci that generated PCR products with expected sizes on agarose gel were assessed for polymorphisms by high-resolution capillary electrophoresis. PCR products were generated by Touchdown PCR with fluorescently labeled M13 (\u201321) using SYBR Green method essentially as described Gray correlation analysis software (V2.1) was used to generate correlation coefficient between gene expression levels and oil content or fatty acid composition Na), effective number of alleles , expected heterozygosity and observed heterozygosity (Ho) were estimated based on the capillary electrophoresis data with POPGENE version 1.31 PIC) values at each locus were calculated as described The number of alleles was detected by capillary electrophoresis of the PCR-amplified products. Genetic parameters including the number of alleles in tung oil extracted from the seeds. We therefore analyzed the fatty acid profiles of mature seeds from all 41 tung tree accessions. GC typically identified 7 fatty acid peaks in tung oil corresponding to palmitic acid (16\u22360), stearic acid (18\u22360), oleic acid (18\u22361), linoleic acid (18\u22362), linolenic acid (18\u22363), \u03b1-eleostearic acid (18\u22363) and \u03b2-eleostearic acid and high 28S:18S rRNA ratio (close to 2.0) in the RNA preparations .The pooled RNA from the three seed stages were used to construct cDNA library for better representation of the whole seed developmental stages. Sequences of the complete cDNA library were assembled into 81,805 unigenes with a mean length of 945 bp . These uMIcroSAtellite tool was used to screen the types of microsatellites from the unigene dataset obtained from tung tree seeds. A total of 6,366 SSRs in 5,404 unigenes contained di-, tri-, tetra-, penta- or hexa-nucleotide repeats . They reThe most abundant SSR motif was (AG/CT), which accounted for 31.3% of the total SSR motif . Other aAfter eliminating undesirable unigenes and avoiding duplications of those published SSRs Capillary electrophoresis is more accurate to estimate the sizes of DNA molecules than agarose gel electrophoresis. The positively identified 56 loci by agarose gel electrophoresis were used for polymorphism analysis by capillary electrophoresis. V. fordii accessions per locus ranged from two to ten, the expected heterozygosity (eH) per locus ranged from 0.140 to 0.692 and the polymorphism information content (PIC) per locus ranged from 0.134 to 0.624 (PIC>0.5) and five were moderate polymorphic (0.2548 hours). A third sample was drawn when infection was diagnosed (Tx). We evaluated platelet activation by measuring the expression of P-selectin (CD62P) and fibrinogen binding on the cell surface before and after stimulation with major platelet agonists through flow cytometry. Clinical scores were obtained at admission.n = 18) presented with significantly higher platelet fibrinogen binding at T1 compared with patients who did not get infected . At T1, ROC AUC for association of basal fibrinogen binding with the occurrence of sepsis was 0.79 (95% CI: 0.68 to 0.89). Elevated basal CD62P expression level was associated with increased 90-day mortality ). Kaplan-Meier survival curves illustrated that mortality was significantly higher after stratification based on T1 basal CD62P level . Multivariate logistic regression analysis using clinical scores indicated that addition of CD62P level or of bound fibrinogen level significantly improved prediction of mortality and sepsis , respectively.Patients who developed sepsis (Predisposition to severe infection in selected critically ill medico-surgical adults can be identified on day 1 of admission based on circulating basally activated platelets. Levels of activated platelets may add incremental prognostic information to clinical scoring."} +{"text": "Superparamagnetic iron oxide nanoparticles (SPIONs) are used as T2 magnetic resonance (MR) contrast agents. Nanorods (NRs) offer an interesting alternative to the more widely used nanospheres as they have shown to offer enhanced T2 relaxivities. The combination of MRI with nuclear imaging modalities such as positron emission tomography (PET) or single photon emission computed tomography (SPECT) increases the data available from a single diagnostic scan . Radiolabelling of SPIONs allows high sensitivity nanoparticle biodistribution data which can both aid in future construct design and be used directly for precise liver lesion imaging. In this work we report the synthesis and characterisation of silica shell iron oxide NRs functionalised with varying ratios of polyethylene glycol (PEG) and the tetraazamacrocyclic chelator, DO3A. Direct and facile radiolabelling of the constructs with the radioisotope gallium-68 (t1/2 = 68 min) proceeded with quantitative radiochemical yields in 15 min and no evidence of radioisotope dissociation was observed after 3 h in both serum and in competition with apo-transferrin. Interestingly, it was observed that neither the radiolabelling process nor stability in vitro or in vivo was compromised by the absence of the bifunctional chelating moiety. Consequently silica shell NRs with 100 % PEG coating were evaluated for potential use as SPECT/MR imaging agents; direct radiolabelling with technetium-99m (t1/2 = 6.02 h) proceeded with analogous radiochemical yields and stabilities. In vivo imaging studies showed rapid liver uptake with high T2 contrast, demonstrating the application of silica shell iron oxide NRs as bimodal PET/MR and SPECT/MR liver imaging agents. Preliminary magnetic hyperthermia evaluation indicates the potential future use of the constructs developed as multimodal theranostic agents."} +{"text": "Innominate artery cannulation is indicated in operations for acute and chronic aortic disease and also in case of porcelain aorta or reoperations. We routinely used innominate artery cannulation in 81 elective operations which included ascending aorta replacement.Aim of our study is to determine whether routine innominate artery cannulation is safe and effective for elective surgical procedures including ascending aorta replacement.We excluded from the study all patients who underwent emergent or urgent operation for acute aortic syndrome, any aortic surgery distal to innominate artery and patients who had other than innominate artery cannulation . The final cohort was consisted of 81 patients who underwent elective ascending aorta replacement alone or with concomitant procedures such as aortic root replacement, coronary or/and valve surgical interventions, which were performed only with innominate artery cannulation. Open distal anastomosis after clamping of the aortic arch branches and selective antegrade cerebral perfusion with a flow rate of 10 ml/kg/min, was performed in all patients. 43 peri-operative variables have been investigated.The operations that have been performed are categorized into ascending aorta replacement with interposition graft , Bentall procedure , Bentall plus coronary artery bypass surgery , ascending aorta replacement with interposition graft plus aortic valve replacement , ascending aorta replacement with interposition graft plus coronary artery bypass surgery and ascending aorta replacement with interposition graft plus aortic valve replacement plus coronary artery bypass surgery . In hospital mortality rate was 1.2% (n = 1). Three patients (3.7%) had postoperative stroke, 2 had transient ischemic attack (2.5%) and 7 developed cognitive dysfunction (8.6%). Average antegrade cerebral perfusion period was 20 minutes at a temperature of 29.4 oC. In three cases bilateral (left common artery) antegrade cerebral perfusion was also used.Innominate artery cannulation performed for elective surgical procedures including ascending aorta replacement is safe and effective. It poses low mortality rate and a low risk of neurological events and cognitive dysfunction."} +{"text": "Congenital heart disease (CHD) is associated with brain dysmaturation, increased risk of perioperative white matter (WM) injury and neurodevelopmental delay (1). Fetal Doppler studies have shown altered cerebrovascular flow dynamics in CHD, suggesting \u2018brain-sparing physiology'. We sought to determine whether postnatal cerebral WM microstructural abnormality relates to abnormal fetal hemodynamics.This prospective IRB approved study included 15 fetuses with CHD and 25 normal fetuses at 36 gestational weeks, who underwent phase-contrast MRI using metric optimized gating and T2-mapping for MR oximetry in the major vessels, according to our previously published technique (2). Neonatal MRI was performed at 5 days (SD 5 days) including diffusion tensor imaging and axial T2W-FSE. Apparent diffusion coefficient (ADC) values were measured in the centrum semiovale (CSO), frontal and parietal deep WM, and mean cerebral WM-ADC was calculated. Visual scoring of WM was performed using a 0 - 6 scale (number of T2-hyperintense regions).2), VO2 and CVO2 were significantly lower in the CHD group . Although mean SVC flow was higher in CHD fetuses, the difference was not statistically significant. However, there was a moderately strong correlation between SVC flow and mean cerebral WM-ADC and VO2 Figure . ADC was) Figure and 1c.FWe conclude that neonatal WM abnormalities in CHD are due to reduced fetal oxygenation. The correlation between more severe WM abnormality and elevated SVC flow, a known marker of acute fetal hypoxia (the \"brain sparing\" response) is further evidence that WM changes are driven by reduced cerebral oxygenation in utero.This study was funded through an operating grant from the CIHR and a PHN New Scholar Award (NIH)."} +{"text": "FBXW7 is deleted and mutated in many different types of human cancers. FBXW7 primarily exerts its tumor suppressor activity by ubiquitinating different oncoproteins including mTOR. Here we used gene transcript profiling to gain a deeper understanding of the role of FBXW7 in tumor development and to determine the influence of mTOR inhibition by rapamycin on tumor transcriptome and biological functions. In comparison to tumors from p53 single heterozygous (p53+/\u2212) mice, we find that radiation-induced thymic lymphomas from Fbxw7/p53 double heterozygous (Fbxw7+/\u2212p53+/\u2212) mice show significant deregulation of cholesterol metabolic processes independent of rapamycin treatment, while cell cycle related genes were upregulated in tumors from placebo treated Fbxw7+/\u2212p53+/\u2212 mice, but not in tumors from rapamycin treated Fbxw7+/\u2212p53+/\u2212 mice. On the other hand, tumors from rapamycin treated Fbxw7+/\u2212p53+/\u2212 mice were enriched for genes involved in the integrated stress response, an adaptive mechanism to survive in stressful environments. Finally, we demonstrated that the Fbxw7 gene signatures identified in mouse tumors significantly overlap with FBXW7 co-expressed genes in human cancers. Importantly these common FBXW7 gene signatures between mouse and human are predictive for disease-free survival in human colon, breast and lung adenocarcinoma cancer patients. These results provide novel insights into the role of FBXW7 in tumor development and have identified a number of potential targets for therapeutic intervention.The tumor suppressor gene FBXW7 encodes an F-box protein, mutated and deleted in cancers from a wide spectrum of human tissues, such as bile duct [FBXW7 in human cancers is approximately 6% [Fbxw7 gene leads to embryonic lethality, but heterozygous mice develop normally [Fbxw7 and p53 show acceleration of tumor development after radiation exposure [The human tumor suppressor gene ile duct , blood [ile duct \u20135, bone ile duct , brain [ile duct , breast ile duct , colon [ile duct , endometile duct , stomachile duct , lung [1ile duct , ovary [ile duct , pancreaile duct , and proile duct . The ovenormally , 18. Altexposure . Mechaniexposure , 21, c-Jexposure , CCNE [2exposure \u201325, diffexposure \u201328, Auroexposure , 29, andexposure .FBXW7 gene may result in impaired degradation of multiple targets and their consequent accumulation, which may cooperatively contribute to tumor development. Our previous studies using mouse models showed that temporal pharmacological inhibition of the mTOR pathway was sufficient to suppress the tumor development contributed by Fbxw7 loss, suggesting that the Fbxw7-mTOR pathway plays a major role in this radiation-induced carcinogenesis mouse model [How does the decrease in FBXW7 function result in tumor development? Deletion or mutation of the se model . In thisFbxw7 significantly reduced radiation-induced tumor latency in p53+/\u2212 mice [FBXW7 alleviates the inhibitory effect on mTOR signaling resulting in activation of the mTOR pathway. Our previous study showed that temporal inhibition of mTOR by rapamycin significantly delayed tumor development in Fbxw7/p53 double heterozygous (Fbxw7+/\u2212p53+/\u2212) mice [n = 11) and vehicle (n = 13) treated Fbxw7+/\u2212p53+/\u2212 mice or from vehicle treated p53 single heterozygous (p53+/\u2212) mice (n = 8) mice (n = 8). When we combined the transcript data from this tumor set with the transcript data of thymic lymphomas from vehicle treated p53+/\u2212 mice (n = 8) significant fold-changes were found for FBXW7, SREBF2 and HMGCS1 and significant enrichment of pathways involved in cholesterol metabolism mice . To detee Figure . Interese Figure . We vali1 Figure confirmim Figure . SREBF1 se liver , 33.Fbxw7+/\u2212p53+/\u2212 mice consistent with the tumor suppressor functions of Fbxw7 in cellular growth and division pathways. Interestingly, we observed significant enrichment of spingosine-1-phosphate (S1P) signaling . These results indicate that radiation induced tumors that arise in a Fbxw7+/\u2212p53+/\u2212 background exhibit decreased latency mediated at least in part through increased cell proliferation mechanisms, and temporal rapamycin treatment delayed tumor development through the inhibition of cell proliferation pathways. We next addressed how rapamycin treatment after radiation exposure can alter tumor transcript profiles.We next analyzed tumor transcript level changes that were unique to the vehicle treated g Figure . Cross-tFbxw7+/\u2212p53+/\u2212 mice. We found 976 transcripts differentially expressed in rapamycin treated Fbxw7+/\u2212p53+/\u2212 thymic lymphomas when compared to p53+/\u2212 tumors , breast and lung cancer , respectively had significantly increased survival compared to patients with a low score (bottom 25%) revealed that ectively , 37. Inty Figure . Given tvailable \u201340. We d) Figure . Taken tFBXW7 significantly reduced tumor latency and that temporal inhibition of mTOR pathway was sufficient to suppress tumor development contributed by Fbxw7 loss in this model, suggesting that Fbxw7-mTOR pathway plays a major role. Here we used transcript profiling of radiation induced thymic lymphomas to investigate the molecular signaling pathways that contribute to tumorigenesis in Fbxw7 heterozygous mice and their dependence on mTOR. We observed significant enrichment of genes regulated by SREBF1 and 2 . The FBXW7-SREBF axis has been proposed in mouse models of non-alcoholic fatty liver disease [Fbxw7 heterozygous mice treated with rapamycin compared to vehicle control treated tumors can cause the accumulation of unfolded proteins in the ER resulting in the unfolded protein response (UPR). Challenges include anoxia, amino acid starvation, glucose deprivation and oxidants. The initial response to this stress is to restore homeostasis promoting cell survival by increasing expression of genes involved in protein folding and degradation of misfolded proteins. When the initial response fails, prolonged ER stress can result in apoptosis. Our study shows upregulation of ATF4, one of the mediating transcription factors of the UPR. Tumors often grow under suboptimal conditions in an environment low in oxygen and nutrients. Thus, it may not be surprising that adaptation to ER stress is a determinant of tumor progression [Fbxw7 wild type, but otherwise isogenic mice concurrently treated with rapamycin did not exhibit enrichment of this response suggesting cross-talk between FBXW7 and mTOR and ER stress. Recently, bidirectional cross talk between mTOR and the UPR have been suggested with mTOR functioning both upstream and downstream of ER stress signals [In gression , 49. It signals \u201352. Inhi signals , 54. Ourp53+/\u2212 and p53+/\u2212 Fbxw7+/\u2212 mice were generated by crossing p53\u2212/\u2212 mice with Fbxw7+/\u2212 mice. At 5 weeks of age, mice were exposed whole-body to a single dose of 4 Gy X-ray irradiation. Mice were divided randomly into two groups and treated with rapamycin or placebo [All animal experiments were performed at Lawrence Berkeley National Laboratory and the study was carried out in strict accordance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The animal use protocol was approved by the Animal Welfare and Research Committee of the Lawrence Berkeley National Laboratory. Detailed methods were described previously . Briefly placebo .Total RNA quality and quantity were determined using Agilent 2100 Bioanalyzer and NanoDrop ND-1000. Agilent SurePrint G3 Mouse GE 8 \u00d7 60 K Microarrays were used according to the manufacturer's protocol . All processes were performed by Ambry Genetics . Microarray data have been deposited at NCBI GEO (accession number: GSE71975).Fbxw7+/\u2212p53+/\u2212 mice and p53+/\u2212 mice were identified by the unpaired Student's t-test with a p-value cut-off of less than 0.05 and a fold change criteria of more than 1.3. Gene lists were annotated with biological functions using Ingenuity Pathway Analysis (IPA), KEGG pathway analysis and DAVID GO gene ontology .Data normalization was performed using GeneSpring GX12.5 (Agilent Technologies). Signal intensities for each probe were normalized to the 75th percentile without baseline transformation. Genes that were differentially expressed in thymic lymphomas between vehicle and rapamycin treated FBXW7 in colon, lung and breast cancer were obtained from the TCGA study at cBioPortal. The overlap between FBXW7 co-expressed genes and the set of genes deregulated in Fbxw7+/\u2212/p53+/\u2212 mouse tumors was used to test for association with disease-free survival. Gene expression microarray datasets of colon, lung and breast cancer for which disease-free survival was available were downloaded from the NCBI GEO website. Kaplan-Meier plots were constructed and a long-rank test was used to determine differences among disease free survival according to a score of FBXW7 gene signature for each patient as the sum of the normalized expression intensities of all genes in each signature.The list of genes that are co-expressed with"} +{"text": "PTPN22 SNPs as non-HLA genetic variants to be associated with susceptibility to autoimmune diseases. Based on these, we looked for association of genetic variants of STAT 3, STAT 5B and GWAs identified PTPN22 with RHD in North Indian population.Rheumatic heart disease (RHD) is an inflammatory, autoimmune disease, occurring as a consequence of group A streptococcal infection complicated by rheumatic fever (RF). Cytokines are important mediators of inflammatory and immune responses. JAK-STATs have been demonstrated to be critical elements in signaling by certain families of cytokines. GWAS has identified STAT3 CG and GG genotypes were significantly associated with RHD (p=0.024 & p=0.027 respectively), STAT5b CT&TT genotypes were significantly associated with RHD (p=0.001 & p=0.002 respectively) while both the SNPs of PTPN22 gene did not show any association with RHD. Further categorization of RHD patients into mitral valve disease (MVD) and combined valve disease (CVD) subgroups revealed that STAT3 CG&GG genotypes were associated with MVD and STAT5b CT&TT genotypes were also associated with both MVD&CVD.This case-control study included 400 RHD patients and 200 controls. The polymorphisms were identified using RFLP/Taqman probes. Statistical analysis was performed by using SPSS. We observed that STAT3 &STAT5b gene polymorphisms may play an important role in the pathogenesis of RHD but GWAS identified PTPN22 SNPs may not be associated with susceptibility of RHD."} +{"text": "Nosocomial infection is becoming a leading problem in medical practitioners now-a-days placing an extra burden on individual patients worldwide. Nosocomial urinary tract infection caused by multidrug resistant (MDR) pathogens is a major threat of the patients in developing country which are increasing numbers in Nepal.The aim of this study was to determine the etiology of nosocomial urinary tract infection caused by multidrug resistant bacterial pathogens.A total of one hundred twenty two bacterial strains isolated from the patients diagnosed of nosocomial urinary tract infection were studied during 2011-2012 at Tribhuvan University Teaching Hospital (TUTH). Antibiotic sensitivity test was determined by modified Kirby Bauer Disc Diffusion method as described by Clinical and Laboratory Standards Institute (CLSI). Data were analysed by using SPSS version 17.0 software.Escherichia coli 51(41.8%) was found to be more predominant which was followed by Acinetobacter calcoaceticus baumannii (Acb) complex 19(15.6%), Klebsiella pneumoniae 11(9%) Enterococcus spp. 18(14.8%) and Staphylococcus aureus 11(9%). Of the total isolates, 74.6% was MDR which is much higher in Klebsiella pneumoniae 100% which was followed by Escherichia coli 90.1%.Nosocomial urinary tract infection was caused by The emergence of MDR bacterial strains causing nosocomial urinary tract infection are increasing in number. The high prevalence of MDR has demanded the special attention to the management of such patients and prevention of dissemination of such strains into hospitals.None declared."} +{"text": "Stagonospora nodorum, resulted in the isolation of three aromatic compounds (1\u20133), including a novel natural butyrophenone, (+)-4\u2032-methoxy-(2S)-methylbutyrophenone (1), and two known polyketides, alternariol (2) and (\u2212)-(3R)-mellein methyl ether (3). Fungal aromatic compounds comprise an important and structurally diverse group of secondary metabolites. Several genome sequencing projects revealed many putative biosynthetic gene clusters of fungal aromatic compounds, but many of these genes seem to be silent under typical laboratory culture conditions. To gain access to this untapped reservoir of natural products, we utilized chemical epigenetic modifiers to induce the expression of dormant biosynthetic genes. As a result, the concomitant supplementation of the histone deacetylase inhibitors suberoylanilide hydroxamic acid (500 \u03bcM) and nicotinamide (50 \u03bcM) to the culture medium of a fungal pathogen,"} +{"text": "IFN\u03b1/\u03b2R1) null hNTCP-Tg mice indicated that in addition to induction of type I IFN response, HDV infection was also associated with up-regulation of novel cellular genes that may modulate HDV infection. Our work has thus proved the concept that NTCP is a functional receptor for HDV infection in vivo and established a convenient small animal model for investigation of HDV pathogenesis and evaluation of antiviral therapeutics against the early steps of infection for this important human pathogen.Hepatitis D virus (HDV) is the smallest virus known to infect human. About 15 million people worldwide are infected by HDV among those 240 million infected by its helper hepatitis B virus (HBV). Viral hepatitis D is considered as one of the most severe forms of human viral hepatitis. No specific antivirals are currently available to treat HDV infection and antivirals against HBV do not ameliorate hepatitis D. Liver sodium taurocholate co-transporting polypeptide (NTCP) was recently identified as a common entry receptor for HDV and HBV in cell cultures. Here we show HDV can infect mice expressing human NTCP (hNTCP-Tg). Antibodies against critical regions of HBV envelope proteins blocked HDV infection in the hNTCP-Tg mice. The infection was acute yet HDV genome replication occurred efficiently, evident by the presence of antigenome RNA and edited RNA species specifying large delta antigen in the livers of infected mice. The resolution of HDV infection appears not dependent on adaptive immune response, but might be facilitated by innate immunity. Liver RNA-seq analyses of HDV infected hNTCP-Tg and type I interferon receptor 1 . HDV is the smallest virus known to infect human. With co-infection of its helper hepatitis B virus (HBV), viral hepatitis D is considered as the most severe form of viral hepatitis. No specific anti-HDV drugs are available; antivirals against HBV do not ameliorate hepatitis D. We report mice expressing a human bile acids transporter sodium taurocholate co-transporting polypeptide (NTCP) in the liver support HDV infection, providing a useful model for studying antivirals against HDV and understanding how the simplest virus interacts with a host Hepatitis D virus (HDV) is the smallest virus known to infect humans with a single\u2014stranded, circular RNA genome of about 1.7 kilobases in length. It is enveloped by surface proteins from its helper hepatitis B virus (HBV) and undergoes a unique replication cycle via an intermediate, antigenomic RNA . Prevalein vitro. For instance, exogenous expression of human NTCP (hNTCP) rendered HDV infection of multiple mammalian cell lines, while successful HBV infection has only been achieved in hNTCP\u2014expressing human hepatoma cells dCTP labeling method as reported by Casey et al [Liver tissue RNA was extracted using TRIzoley et al or quant+/-) mice, and HDV-inoculated mice including three hNTCP+/-, three hNTCP+/-/IFN\u03b1/\u03b2R-/-, and three wild-type mice. Mice were inoculated on day 9 after birth, and sacrificed 6 days after the inoculation. RNA-seq was performed using Illumina Genome Analyzer IIx system. Sequence data was deposited at Sequence Read Archive (SRA) of the NCBI under BioProject PRJNA236433. See RNA-seq analysis was conducted using the total RNA of livers from 3 mock-inoculated hNTCP transgenic (hNTCPS1 Table(XLSX)Click here for additional data file.S1 Text(DOCX)Click here for additional data file.S1 Fig(TIF)Click here for additional data file.S2 Fig(TIF)Click here for additional data file.S3 Fig(TIF)Click here for additional data file.S4 Fig(TIF)Click here for additional data file.S5 Fig(TIF)Click here for additional data file."} +{"text": "Zoysia japonica Steud.) is commonly found in temperate climate regions and widely used for lawns, in part, owing to its uniform green color. However, some zoysiagrass cultivars accumulate red to purple pigments in their spike and stolon tissues, thereby decreasing the aesthetic value. Here we analyzed the anthocyanin contents of two zoysiagrass cultivars \u2018Anyang-jungji\u2019 (AJ) and \u2018Greenzoa\u2019 (GZ) that produce spikes and stolons with purple and green colors, respectively, and revealed that cyanidin and petunidin were primarily accumulated in the pigmented tissues. In parallel, we performed a de novo transcriptome assembly and identified differentially expressed genes between the two cultivars. We found that two anthocyanin biosynthesis genes encoding anthocyanidin synthase (ANS) and dihydroflavonol 4-reductase (DFR) were preferentially upregulated in the purple AJ spike upon pigmentation. Both ANS and DFR genes were also highly expressed in other zoysiagrass cultivars with purple spikes and stolons, but their expression levels were significantly low in the cultivars with green tissues. We observed that recombinant ZjDFR1 and ZjANS1 proteins successfully catalyze the conversions of dihydroflavonols into leucoanthocyanidins and leucoanthocyanidins into anthocyanidins, respectively. These findings strongly suggest that upregulation of ANS and DFR is responsible for tissue-specific anthocyanin biosynthesis and differential pigmentation in zoysiagrass. The present study also demonstrates the feasibility of a de novo transcriptome analysis to identify the key genes associated with specific traits, even in the absence of reference genome information.Zoysiagrass ( Zoysia japonica Steud.) is a perennial creeping grass commonly found in Southeast and East Asia and Australia ((TIF)Click here for additional data file.S19 FigThe guide bar indicates 0.5 mm.(TIF)Click here for additional data file.S1 File(DOC)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(DOCX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S6 Table(DOCX)Click here for additional data file.S7 Table(DOCX)Click here for additional data file.S8 Table(DOCX)Click here for additional data file.S9 Table(DOCX)Click here for additional data file.S10 Table(DOCX)Click here for additional data file."} +{"text": "Endothelial dysfunction plays a key role in sepsis physiopathology, leading to multiorgan failure and death. Circulating endothelial cells (CECs) and circulating endothelial progenitor cells (CEPCs) may be elevated in septic syndrome as a result of endothelial damage. Previous studies have shown conflicting results and their role in septic syndrome remains unclear , 2.Compare the number of circulating endothelial cells (CECs) and circulating endothelial progenitor cells (CEPCs) in septic and non-infectious systemic inflammatory response syndrome (SIRS) and controls; and evaluate their association with outcome.Blood samples from patients affected with septic syndrome and SIRS were prospectively collected within the first 24 hours and after 96 hours of symptoms onset. Patients with malignancies were excluded. A healthy control group was included (n= 15). CECs and CEPCs count were determined by flow cytometry and were calculated to obtain their absolute number per 1 mL of whole blood.Fifty-seven patients were included in the study: sepsis (n = 20), severe sepsis (n = 10), septic shock (n = 16) and non-infectious SIRS (n = 11). Sixty-five percent were male and median age was 68 years. Most of the patients presented non-surgical pathologies (93%). Patients of all groups presented higher number of CECs and CEPCs than healthy volunteers in baseline (p < 0.001 and 0.034 respectively). A positive correlation was found between CECs and CEPCs count in the first day in all groups (p < 0.001). CECs and CEPCs count decreased significantly in patients with severe sepsis/septic shock at 96 hours (p = 0.034). There were not significant differences between CECs and CEPCs count either between the different groups or survivors and non-survivors. We did not find any correlation between the number of CECs or CEPCs and severity or organ dysfunction at any time point.Patients with SIRS of infectious and non-infectious origin present an increase of CECs and CEPCs count. However, different septic syndrome and non-infectious SIRS do not present differences in these count. Finally, CECs and CEPCs count seem not associated with survival or illness severity in these patients.FIS PI12/00832, Instituto de Salud Carlos II y cofinancaido con Fono Europeo de Desarrollo Regional (FEDER), Uni\u00f3n Europea, Una manera de hacer Europa."} +{"text": "To develop and evaluate a non-invasive 3D model of Myocardial Wall Stress (MWS) in subjects with severe aortic valve stenosis (AVS).We studied 20 subjects (71\u00b110 years) with severe AVS including 19 with a preserved ejection fraction (EF) and 14 elderly controls (59\u00b16 years). All subjects underwent within 4 hours, transthoracic echocardiogram (TTE) and cardiac magnetic resonance (CMR) as well as applanation tonometry of the carotid artery immediately after CMR examination. CMR included an ECG gated cine SSFP acquisition of left ventricular (LV) short axis slices, positioned regularly and without inter-slice gap between base and apex. Carotid tonometry, by adjusting by the mean brachial pressure obtained during CMR acquisition is a reliable measurement of central aortic blood pressure. 3D MWS can provide a LV afterload estimate which is well known to be strongly related to EF, except in case of depressed contractility. Evaluation of 3D MWS relied on the combination of: 1) a geometrical factor (GF), estimated according to myocardial thickness and LV cavity radius, while accounting for the 3D curvature of the LV, and 2) LV peak systolic pressure provided by tonometric measurement and Doppler maximal transvalvular pressure gradient.For all patients, TTE revealed the presence of severe AVS according to ESC criteria . When compared to 3D MWS evaluation of controls, GF indicated a significant decrease in patients with AVS whereas 3D MWS remained equivalent between the two groups. These data reflect LV adaptation to pressure overload, leading to an overall normalization of MWS in severe AVSwith preserved EF. Furthermore, our 3D model of MWS is strongly related to EF , reflecting the robustness of this non-invasive method based on CMR and applanation tonometry.This 3D non-invasive evaluation of MWS based on CMR and carotid tonometry was able to characterize LV adaptation to pressure overload in severe AVS. Since, MWS provides a LV afterload estimate, which is strongly related to EF, except in case of depressed contractility, future studies should be performed to reveal the particular usefulness of such non-invasive 3D MWS for prediction of recovery after aortic valve replacement in subjects with low EF.This study was funded by Assistance Publique dex H\u00f4pitaux de Paris."} +{"text": "Rib fracture nonunion represents failure of normal fracture healing. Although randomized controlled trials have demonstrated benefit to acute rib stabilization, the role of open reduction and internal fixation (ORIF) for symptomatic nonunion fractures is unknown and limited to case reports.We review and report our recent consecutive series of ORIF for symptomatic nonunion rib fractures.All consecutive patients who underwent rib stabilization for symptomatic nonunion between 2010 and 2014 were retrospectively reviewed. Indications included persistent fracture on imaging accompanied with pain. Outcomes were analysed on 1) radiographic criteria including postoperative chest X-ray at 2 weeks, and CT scans at 3 and 6 months and 2) patient symptoms.Eight patients underwent non-union rib stabilization of 1 to 4 ribs during the study period. Median age was 56 years . Mean BMI was 31.8 and median interval from index injury to rib fracture surgical repair was 14.5 months . 75% of this cohort used tobacco chronically within the 3 years preceding repair. One patient underwent stabilization with ORIF alone and the remaining 7 patients underwent ORIF plus autologous bone grafting. There was no operative mortality. Median length of stay was 3.5 days . Complications included 2 surgical site infections treated with surgery and 1 patient with pneumonia requiring antibiotics. At a mean follow up of 9.8 months (range 1-27), all patients reported symptomatic improvement. Radiographic healing was present in 100%.Rib stabilization with bone grafting may be a successful alternative in the management of symptomatic non-union rib fractures. With increased experience with this thoracic surgical option, earlier intervention in select cases may permit more rapid symptom control and better outcomes."} +{"text": "Cicer spp.) comprising of cultivated and wild species, six abiotic stress treatments , and five diverse tissues . The geNorm, NormFinder and RefFinder algorithms used to identify stably expressed genes in four sample sets revealed stable expression of UCP and G6PD genes across genotypes, while TIP41 and CAC were highly stable under abiotic stress conditions. While PP2A and ABCT genes were ranked as best for different tissues, ABCT, UCP and CAC were most stable across all samples. This study demonstrated the usefulness of new generation reference genes for more accurate qPCR based gene expression quantification in cultivated as well as wild chickpea species. Validation of the best reference genes was carried out by studying their impact on normalization of aquaporin genes PIP1;4 and TIP3;1, in three contrasting chickpea genotypes under high vapor pressure deficit (VPD) treatment. The chickpea TIP3;1 gene got significantly up regulated under high VPD conditions with higher relative expression in the drought susceptible genotype, confirming the suitability of the selected reference genes for expression analysis. This is the first comprehensive study on the stability of the new generation reference genes for qPCR studies in chickpea across species, different tissues and abiotic stresses.Quantitative Real-Time PCR (qPCR) is a preferred and reliable method for accurate quantification of gene expression to understand precise gene functions. A total of 25 candidate reference genes including traditional and new generation reference genes were selected and evaluated in a diverse set of chickpea samples. The samples used in this study included nine chickpea genotypes ( GAPDH), actin (ACT) tubulin (TUB) cyclophilin (CYP), elongation factor (EF1) and ribosomal RNA (18S and 28S rRNA) have been widely used as reference genes in normalization of qPCR data. However, several studies have revealed that the expression levels of many commonly used housekeeping genes vary across tissues, treatments and species [Gene expression studies have become extremely important to obtain insights into gene function and to understand the molecular mechanisms. Among various techniques used for gene expression studies, quantitative Real-Time PCR (qPCR) has become the most important and reliable method owing to its accuracy and high-throughput analysis . While afactor EF and ribo species \u20135. Hence species , 6. Sinc species , 8. More species , NormFin species , BestKee species and comp species have beeArabidopsis [Brassica juncea [Brassica napus [Coffea species [Gossypium hirsutum [Oryza sativa [Solanum tuberosum [Solanum lycopersicum [Triticum aestivum [Vitis vinifera [Zea mays [Atropa belladonna [Caragana korshinskii [Panicum virgatum [Pennisetum glaucum [Phalaenopsis [Populus euphratica [Saccharum officinarum [Glycine max [Arachis hypogaea [Vicia faba [Pisum sativum [Lens culinaris [Cicer arietinum [Studies on the evaluation of reference genes have been carried out in several plant species such as bidopsis , Brassica juncea , Brassicca napus , 16, Cof species , Gossypihirsutum , Oryza sa sativa , Solanumuberosum , Solanumaestivum , Vitis vvinifera , and ZeaZea mays . More relladonna , Caraganshinskii , Panicumvirgatum , Pennise glaucum Phalaenoaenopsis Populus phratica and Saccicinarum . Among tcine max and Araccia faba , in Pisu sativum in Lens ulinaris and in Crietinum . Since, nutraceutical because of its high nutritional value [Ascochyta blight, Fusarium wilt and pod borer). With chickpea genome sequencing allowing high-powered functional genomics studies to proceed [Chickpea is an important food legume of the semi-arid tropical (SAT) regions of the world, known to be a al value . Despite proceed , these cABCT), alcohol dehydrogenase class-3 (ADH3), calcium-dependent protein kinase 4 (CDPK4), clathrin adaptor complexes medium (CAC), cyclophilin (CYP), eukaryotic elongation factor 1-alpha (ELF1a), eukaryotic elongation factor 1-beta (ELF1b), galactose oxidase/kelch repeat superfamily protein (FBOX), glucose-6-phosphate dehydrogenase (G6PD), glyceraldehyde3-phosphate dehydrogenase (GAPDH), heat shock cognate protein 80 (HSP80), translation initiation factor IF-3 (IF3), eukaryotic initiation factor 4A-15 (IF4a), peroxin4 (PEX4), protein phosphatase2A subunit A3 (PP2A), pentatrico peptide repeat superfamily protein (PPR), s-adenosyl methionine decarboxylase (SAMDM), SAND-family protein (SAND), F-box protein SKIP16-like (SKIP16), TIP41-like protein (TIP41), un-characterized conserved protein UCP022280 (UCP), unknown protein (UNK), ubiquitin-protein ligase 7 (UPL7), vacuolar protein sorting-associated protein 53 homolog (VPS) and yellow leaf specific protein 8 (YLS8). The expression stability of these genes was evaluated across gene pools, different tissues and stress treatments using geNorm, NormFinder and RefFinder algorithms. Further, we have selected the best and least stable reference genes from the all samples set and validated by normalizing the expression levels of two aquaporin genes PIP1;4 (Plasma membrane intrinsic protein) and TIP3;1 (Tonoplast intrinsic protein) of chickpea. These two aquaporin genes express differentially in susceptible and tolerant genotypes of chickpea under different abiotic stress conditions (Unpublished data). The expression levels of the selected aquaporin genes under vapor pressure deficit (VPD) stress were tested in three chickpea genotypes with different tolerance levels.Keeping this in view, we have evaluated a set of reference genes including traditional (commonly used) and new generation reference genes in a diverse set of biological samples of chickpea, including nine genotypes representing cultivated and wild species across primary, secondary and tertiary gene pools, plant tissues from various developmental stages, and six abiotic stress treatments . To identify the most stable reference gene(s) for normalization of qPCR data, this study evaluated 25 reference genes, including ATP-binding cassette transporter , Hyderabad, India. Chickpea plants were grown in 8-inch pots containing 4.5 kg of black clay soil under glasshouse conditions with 28/20\u00b0C day/night temperature. Nine chickpea genotype representations from cultivated genotypes and wild species across primary, secondary and tertiary gene pools were useArabidopsis and other grain legumes such as soybean, chickpea and peanut followed by an in silico analysis using BLAST tools of the NCBI database. A total of 28 candidate reference genes that showed highly stable expression in previous studies including CAC, FBOX, GAPC2, PEX4, PP2A, PPR, PTB1, SAMDM, SAND, TIP41, UCP, UNK, UPL7 and YLS8 in Arabidopsis [ABCT, CDPK4, IF3 and SKIP16 in soybean [ELF1a, GAPDH, HSP80 and IF4a in chickpea [ACT, ADH3, CYP, ELF1b and G6PD in peanut [VPS gene from chickpea (unpublished) were selected for evaluation. To retrieve the orthologous mRNA and their corresponding DNA sequences in chickpea, mRNA sequences of the Arabidopsis genes, EST sequences of the chickpea, soybean and peanut were used to query databases of the National Center for Biotechnology Information (NCBI) using BLASTX. The retrieved mRNA sequences of the chickpea sequences were used to design PCR primers using Primer3 software [The candidate genes were selected based on a review of studies in model plant bidopsis ; ABCT, C soybean ; ELF1a, chickpea ; ACT, AD2, 200 \u03bcM dNTP and 1 U Taq polymerase . PCR samples were amplified in an Eppendorf Thermal Cycler with an initial denaturation at 95\u00b0C for 5 min followed by 35 cycles of 95\u00b0C for 1 min, 62\u201364\u00b0C for 1 min and 72\u00b0C for 3 min, followed by a final extension at 72\u00b0C for 10 min, and PCR products tested by using 1% agarose gel electrophoresis.The genomic DNA from leaves of chickpea variety JG11 was isolated using NucleoSpin Plant II DNA isolation kit . PCR reactions for all the candidate genes were performed using genomic DNA as template and gene-specific primers designed for qPCR. The PCR was performed in a total volume of 25 \u03bcl using 100 ng of genomic DNA, 200 nM of each primer, 1.5 mM MgCl\u00ae III first strand cDNA synthesis kit and oligodT primer in 25 \u03bcL reaction, according to manufacturer\u2019s protocol. The cDNA preparations were diluted 12-times with nuclease-free water to use as template in qPCR analysis. To confirm the total absence of genomic DNA, cDNA was used as a template for PCR amplification using ADH3 and G6PD primer pairs spanning an intron and PCR was performed as mentioned above.Total RNA was extracted from 100 mg tissue by using NucleoSpin RNA plant kit including in-column DNAse1 treatment. The concentration and purity of all RNA samples was tested using NanoVue plus spectrophotometer at 260/280 nm absorbance, selecting the ones that ranged from 1.8 to 2.0. Integrity of the RNA was tested by using 1.4% agarose gel electrophoresis with SYBR safe DNA gel stain . The total RNA sample (2.5 \u03bcg) was reverse transcribed to cDNA using SuperScriptTM SYBR No-ROX mix, 400 nM of each primer, 1.0 \u03bcL of diluted cDNA and nuclease-free water to make up the final volume. The reaction conditions were set as 2 min at 95\u00b0C (polymerase activation); 40 cycles of 15 s at 95\u00b0C, 30 s at 62\u00b0C with fluorescent signal recording. At the end, a final step of 15 s at 95\u00b0C, 30 s at 58\u00b0C and fluorescence measurement at each 0.5\u00b0C variation from 58\u00b0C to 95\u00b0C in 20 min was included to obtain the melting curve. No-template controls were included for each primer combinations. Pooled and diluted cDNA sample was used in qPCR to check the specificity of all the primer pairs and verified by using 2% agarose gel electrophoresis with SYBR safe DNA gel stain prior to sequencing the amplified products. The sequences of the PCR amplified products of each primer combination were compared with GenBank sequences using the BLASTN algorithm to check the PCR product specificity. For expression profiling, all the cDNA samples were tested in qPCR with each primer pair and three technical replicates performed for each cDNA sample of each biological replicate. The quantitative cycle (Cq) values were recorded using default settings of Real time PCR system where baseline was corrected automatically and threshold value was estimated by setting to noise band mode. Statistical analysis (Mean and CV) of the Cq values was carried out using Microsoft Excel spreadsheet 2010. The PCR efficiency of each primer pair was evaluated by the dilution series method using a pooled cDNA sample of the chickpea variety JG11. The 12-times diluted pooled cDNA sample was taken and 2-fold serial dilutions were carried out. Five serially diluted cDNA samples were used as templates to construct the standard curves of each primer pair, where PCR composition and conditions were same as above. Standard curves were constructed using the RealPlex real time PCR instrument software by using linear regression based on the quantitative cycle (Cq) values for all dilution points in a series. The correlation coefficients (R2) and slope values were obtained from the standard curve, and corresponding PCR amplification efficiencies (E) were calculated using the slope of the calibration curve according to the following equation: E = (10\u22121/slope-1).All qPCRs were carried out in Realplex Real-Time PCR system using SYBR Green in 96 well optical reaction plates sealed with ultra-clear sealing film (Platemax). The PCR reaction was performed in a total volume of 10 \u03bcL containing 5 \u03bcl of 2X SensiFASThttp://www.leonxie.com/referencegene.php) tool that integrates the currently available four major computational programs {geNorm [All experimental samples were divided into different sets based on their nature. The tissue set comprised of leaf, flower, root, seedling and seed of chickpea variety JG11 grown under normal growth conditions (five samples). The abiotic stress treatment set comprised of eight leaf samples of chickpea variety JG11 grown under drought, salt, high VPD, ABA, cold, heat shock, drought control and control. The genotypes sample set comprised of leaf sample of four cultivars and five wild species and TIP3;1 (Tonoplast intrinsic protein) of chickpea were selected as target genes for quantification of gene expression levels under high vapor pressure deficit (VPD) treatment. Three chickpea varieties with contrasting drought tolerance viz. a susceptible variety ICC1882 and two tolerant varieties ICC4958 and JG11 were selected for reference gene validations. Experimental conditions and sample collection followed were same as mentioned above. Gene expression levels of PIP1;4 and TIP3;1 were normalized using the two most stable reference genes (ABCT and UCP) and two least stable genes (CYP and SKIP16) individually and in combination. Relative expressions of these two aquaporin genes in drought stressed leaf samples were estimated by comparing with expression levels of leaf sample collected from GH grown control plants of same variety using the REST [Two aquaporin genes the REST softwareArabidopsis, soybean, peanut and chickpea. Primers designed for 28 candidate reference genes were tested with cDNA of chickpea variety JG11. Since the primer pairs of ACT, GAPC2 and PTB1 genes yielded more than one band with cDNA as template, these genes were eliminated from further analysis. All the remaining 25 primer pairs except for PP2A (The PP2A forward primer was designed from the exon-exon junction region) yielded single PCR product of expected size with the genomic DNA (E) of the candidate reference genes ranged from 0.90 to 1.09 and CDPK the highest (27.55). Amongst all the tested genes, the GAPDH, HSP80 and ELF1b showed higher expression where mean Cq values were below 22. The expression levels of CDPK, PPR, SKIP16, UNK, and FBOX were lower (mean Cq above 26), while the rest of the genes showed intermediate expression levels (mean Cq 22 to 26). The coefficient of variation (CV) values calculated for each reference gene indicated lower gene expression variation in PPR and FBOX and the high variation in the expression of ELF1a (CV: 14.17) , five different tissues and nine genotypes representing cultivated and wild species across the chickpea gene pools. To minimize the variability associated with qPCR analysis, all RNA samples were taken in equal concentration and quality for conversion to cDNA. The expression levels of the all candidate reference genes were individually determined in each of the samples as the quantification cycle value (Cq). A relatively wide range of Cq values across all 20 samples of chickpea was observed for the 25 reference genes, suggesting various levels of transcript abundance of the genes analyzed . The Cq : 14.17) .CAC/ABCT (M = 0.46) corresponding to the most stable expression in all sample set, whereas the M values for SKIP16 and CYP was considerably higher than the rest of the candidate genes. In tissue sample set, VPS and ABCT were most stable and PPR, CDPK were least stable . Under abiotic stress conditions, while TIP41 and G6PD were most stably expressed than all other candidate reference genes tested, ELF1a and PPR were unstable. The genes YLS8 and PEX4 were most stable in the tested genotypes across various gene pools while CYP and SKIP16 were found to be least stable showed UCP as the most stable gene followed by ABCT and G6PD in the all sample set, whereas CYP, SKIP16 and ELF1a genes were least stable among the all candidate reference genes tested. The TIP41 expressed most stably under abiotic stress conditions followed by CAC and PEX4, whereas ELF1a and PPR remained least stable. Across genotypes and species, UCP, G6PD and VPS genes were identified as the most stable, whereas the CYP gene was identified as least stable followed by SKIP16 and ELF1a genes. In the tissue sample set, PP2A, CAC, ABCT were identified as highly stable genes, whereas PPR, CDPK, CYP were among the least stable ones, according to NormFinder analysis.Since the stability value of the best reference gene or the best combination of genes may vary from one experimental set to another, the stability ranks of candidate genes in four sample sets was determined separately to identify the most suitable reference genes using geNorm and NormFinder algorithms. The geNorm program defines a stability measure (M) as the average pairwise variation between a gene and reference genes in a set of samples where genes with the lowest M values have the most stable expression. The lowest M value was observed for the pair t stable . Similart stable showed UUCP, G6PD, CAC and YLS8 reliably expressed in chickpea genotypes across species, ABCT, UCP, CAC and G6PD were most stable in all samples set. Similarly, while PP2A, ABCT, VPS and CAC genes were ranked as top four in differential tissue sample set, TIP41, CAC, G6PD and PEX4 were observed to be highly stable under abiotic stress conditions. The comprehensive ranking revealed that the CYP gene was most unstable gene in all the sample sets, except under abiotic stresses. Various other genes such as ELF1a and GAPDH in abiotic stress, CDPK in differential tissues, SKIP16 in genotypes across species and all samples were also found unreliable in this analysis was higher than 0.15, an ideal normalization will require to include as 0.125 .PIP1;4 and TIP3;1 were investigated in three chickpea varieties, ICC1882, ICC4958 and JG11 under VPD stress. For normalization of the relative expression level of two aquaporin genes, two most stable and two least stable reference genes as evaluated by RefFinder in all sample set were used among all the tested genes must be determined statistically. The stability value of the most suitable reference gene or the best combination of genes may vary from one experimental set to another, and for that reason the choice for reference genes for optimal normalization was made from a set of candidate genes for each experiment and for all of them together. To identify most stable reference gene under different experimental conditions, we grouped the diverse chickpea tissue samples into four sample sets and validated the 25 candidate reference genes using geNorm and NormFinder, two distinct statistical algorithms. Although, several studies have reported differences between the outputs of geNorm and NormFinder , 47, 48,CAC (Arabidopsis homologue: AT5G46630) was identified among the top four reference genes in all four-sample sets, indicating that CAC transcript levels were stable in chickpea under different experimental conditions tested. In this study, the CAC gene was selected as a candidate reference gene based on its highly stable expression in the Arabidopsis [CAC has been reported to be the best reference gene for normalization in tomato [CAC gene is stably expressed in different experimental conditions.Interestingly, new generation candidate reference genes selected for the current study performed better than the traditionally employed reference genes. In particular, n tomato , 17, durn tomato , across n tomato , and at n tomato . Based oUCP, Arabidopsis homologue: AT4G26410) was most stably expressed in different genotypes of chickpea and ranked second in all sample set. Previously, UCP gene was identified as stably expressed gene in Arabidopsis microarray analysis [UCP gene expression has reportedly been moderately stable in Arabidopsis under metal stress [UCP gene was most unstable in vegetative tissues and maturing embryos of rapeseed [ABCT) gene known to play a variety of cellular roles such as auxin transport was also selected for validation in the present study based on the microarray and qPCR validation in soybean, where ABCT has been reported as most suitable for gene expression normalization [ABCT gene was most stable when all samples were analyzed together and second most stable gene in differential tissue sample set, which is in agreement with the study in soybean where the ABCT gene was expressed stably under various abiotic stress conditions [ABCT as the stable reference gene in crop species. Similarly, the TIP41-like family protein coding gene that was selected based on the Arabidopsis microarray data [TIP41 gene stably expressed under different abiotic conditions in desert shrub [TIP41 gene also showed stable and similar expression regulation across Brassica species regardless of experimental conditions [The uncharacterized conserved protein UCP022280 gene under different experimental conditions in peanut [G6PD gene ranked in top four most stably expressed genes across different chickpea genotypes under abiotic stress conditions and all samples set. However, these results are in contrast to the findings in soybean where G6PD gene expression was least stable under developmental stages and different photoperiodic treatments [VPS) gene selected as candidate reference gene in the present study has not been previously explored as reference gene for qPCR analysis, it was identified as low copy gene in the legumes (unpublished data) and subsequently cloned. Interestingly, VPS gene expressed stably in different tissues and showed moderate stability across all samples and genotypes, ranking 5th and 6th respectively, indicating its potential for inclusion as reference gene in expression studies in other crop species.Since our previous studies indicated stable expression of glucose-6-phosphate dehydrogenase that belong to two different aquaporin sub-families (unpublished data) were assessed. Plant aquaporin proteins play major role in water transport through cell membranes and these genes are expressed differentially in different tissues that are also altered under different abiotic stresses in higher plants. The chickpea TIP3;1 gene was significantly up regulated under high VPD conditions in leaf tissues of three chickpea genotypes with relatively higher expression in susceptible genotype ICC1882, when normalized with ABCT and UCP reference genes. These results were in accordance with our previous study in sorghum [TIP genes in leaf tissues was reported under different abiotic stress conditions, but the normalization was obscured when the least stable reference gene(s) (CYP and SKIP16) were used. The validation results clearly indicated the demerits of using unstable reference gene(s) for normalization and confirmed the reference gene stability of the selected candidate genes.To validate the selected candidate reference genes based on RefFinder ranking, expression of two chickpea aquaporin genes selected for the present study performed significantly better than the traditionally employed reference genes that were stably expressed across different experimental conditions. However, the CYP and ELF1a genes that were most unstable should be avoided for use as reference genes in gene expression studies in chickpea. Pairwise variation analysis indicated that two genes are enough for normalization of gene expression data in different sample sets except when all samples were analyzed together, where three genes are required. The validation of selected reference gene by normalizing aquaporin gene expression data further confirmed the stability of the ABCT and UCP as reference genes under high VPD conditions. These results provide information that can ensure more accurate qPCR based gene expression quantification towards chickpea functional genomics.The new generation candidate reference genes Click here for additional data file.S2 Fig(TIF)Click here for additional data file."} +{"text": "Trials and evaluations of interventions for preterm birth prevention have reported many different outcomes resulting in an inability to synthesise results across studies. Our objective was to produce a minimum set of important and critical outcomes (core outcome set) for studies, reviews, evaluations and guidelines on primary prevention of preterm birth.Between May and November 2014 we went through the process of identification and selection of outcomes. All possible outcomes were drawn from systematic reviews, interviews and questionnaires. From this initial list, a selection process was performed using an online 2-round Delphic survey and a consultation meeting. Target stakeholders were approached to contribute in this selection through purposive sampling in relevant networks: patient organisations, midwife networks, Global Obstetrics Network (GONet), the Cochrane collaboration and journal editors from the Core Outcomes in Women\u2019s health (CROWN) initiative.From an initial list of 249 items, 29 outcomes were identified for the process of consensus in the Delphi survey. A total of 228 participants were approached for the Delphi survey of whom 194 (86%) completed the first survey and 174 (89%) the second survey. Responders of both surveys represented all stakeholder groups: parents (n=25), midwives, (n=25), obstetricians (n=54), neonatologists (n=34), methodologists (n=34) and industry (n=2) from 11 low and 16 high resourced countries. The following eleven outcomes were selected as being \u2018critical outcomes\u2019 in the Delphi survey and one more was included after the consultation meeting: [1] maternal mortality; [2] maternal infection or inflammation; [3] gestation age at delivery; [4] offspring mortality; [5] birth weight; [6] offspring early developmental morbidity; [7] offspring late neurodevelopmental morbidity; [8] offspring gastro-intestinal morbidity; [9] offspring infection; [10] offspring respiratory morbidity; [11] prelabor rupture of membranes; [12] harm.We developed a core outcome set for studies on primary prevention of preterm birth. We encourage researchers to start to collect all outcomes that are in this core outcome set.COMET Registration Number: 603.http://www.comet-initiative.org/studies/details/603."} +{"text": "Inferior venecava (IVC) invasion in renal cell carcinoma (RCC) has an incidence of 4-10% and extension in right atrium is <1%, Lung metastasis is most frequent than at other locations. Supradiaphragmatic IVC thrombus with solitary lung metastasis does not affect the prognosis adversely, provided complete resection is achieved.To achieve necessary exposure with complete removal of supradiaphragmatic IVC thrombus, minimise blood loss with support of cardio pulmonary bypass (CPB) with pulmonary metastasectomy in such high risk group of patients.We treated 31 patients of RCC with IVC invasion with lung metastasis in one, after categorising under Mayo Classification. Preoperatively metastasis and thrombus extent were determined by computed tomography. Sternotomy or thoracotomy was performed for supradiaphragmatic IVC thrombus, 2 patients were found unfit to undergo surgery on CPB so operated by off-pump intrapericardial IVC clamping under guidance of transoesophageal echocardiography (TOE) and rest were operated on CPB, pulmonary metastasectomy for solitary lung metastasis was performed in one. Patients followed up every 3 months.Out of 31 patients, 4 had level 0, 27 had IVC thrombus of these 13 had level I, 4 had level II with solitary pulmonary metastatic lesion in one, 3 had level III and 7 had level IV thrombus and out of these 7 patients, 3 were operated on CPB of which 1 died of disseminated intravascular coagulopathy, 2 patients were operated by off-pump intrapericardial IVC clamping under TOE guidance and 2 patients were inoperable and none had recurrence.Role of cardio thoracic surgeon is pivotal in achieving a significantly longer overall survival in patients of metastatic RCC with supradiaphragmatic thrombus with solitary lung metastasis, by complete resection of thrombus on or off CPB with metastasectomy and this multidisciplinary approach may benefit patients awaiting fatal outcome of their natural history."} +{"text": "The aim was to determine whether the degree of enhancement on hepatocellular phase gadoxetic enhanced MRI and the apparent diffusion coefficient (ADC) before and after neoadjuvant chemotherapy could identify pathologic complete responders in colorectal liver metastases (CRLM).In this retrospective study, 22 patients with CRLM were evaluated with gadoxetic acid enhanced MRI before and after chemotherapy. Regions of interest were drawn encompassing metastases on T1W images and ADC map by an expert radiologist to record their average signal intensities (SI) normalised to the SI of paravertebral muscle and the average ADC value. We compared the median ADC value; pre-contrast and hepatocellular phase normalised SI and their percentage change in pathologic complete responders and pathologic non complete responders before and after chemotherapy using Mann-Whitney test. Receiver operating curve characteristics (ROC) of these parameters were determined. A p-value of < 0.05 was deemed statistically significant.All patients received FOLFOX/FOLFIRI based-chemotherapy, while 8 received in addition bevacizumab. There were 37 CRLM at histology, of which 10 showed complete pathological response. There was a significant difference in the median percentage increase in the hepatocellular phase normalised SI of CRLM after neoadjuvant chemotherapy between pathologic complete responders (18%) and pathologic non complete responders (2.5%) (p = 0.04). By ROC analysis, an increase in the median hepatocellular phase normalised SI of 6% after chemotherapy has a sensitivity of 85% (95%CI:55-98%) and specificity of 70% (35-93%) for identifying pathologic complete responders (p=0.03). The other parameters including ADC were not statistically significant.An interval increase in the hepatocellular phase normalised SI of CRLM is associated with pathologic complete response following neoadjuvant chemotherapy."} +{"text": "Foot involvement is commonly described in children with JIA and management of foot disability is necessary to maximize physical function in children. The Juvenile Arthritis Foot disability Index (JAFI) is a questionnaire for assessing self-reported foot-related disability in children and adolescences with arthritis. The JAFI is a valid and reliable survey which is divided into three dimensions; impairment, activity limitations and participation restrictions (Andr\u00e9 2004). However, its ability to measure change in foot related disability has not been established.(a) To evaluate the responsiveness of the JAFI among a cohort of children with JIA undergoing treatment with Intra-articular corticosteroid injections (ICI). (b) To determine the association between JAFI change scores and the Child Health Assessment Questionnaire lower extremity dimension (CHAQ-low) change scores. We hypothesized that (a) the change in JAFI and CHAQ-low scores at 3 months following treatment from a group without foot impairments would demonstrate greater standardized response means (SRM = mean change/ SD change scores) and effect sizes (ES = mean change/ SD baseline scores) compared to scores from children with persistent foot disability. We also hypothesized that (b) JAFI change scores would moderately correlate (r = 0.4) with CHAQ-low change scores.35 children with JIA were consecutively recruited from a large tertiary care medical center pediatric rheumatology clinic. Twenty-eight participants were female; mean age (SD) 11(4) yrs. and mean (SD) disease duration of 4(4) yrs. 66% were diagnosed with polyarthritis. All children had active lower extremity inflammation and were scheduled for ICI in one or both feet. The children or parents (if child <10 yrs.) answered the JAFI and the CHAQ prior to injections, 3 months and 6 months after ICI. Scores from the CHAQ walking, rising, reaching and activity scales were summarized and divided into quartiles (CHAQ-low). To determine the responsiveness of the JAFI, children were separated into 2 groups based on standardized clinical examination, those without foot impairments at 3 months (n = 13) and those with foot impairments (n = 22) and standardized response means and effect sizes were calculated. Spearman correlations were performed to determine the association between JAFI dimensions and CHAQ- low change scores.s) between JAFI dimensions and CHAQ-low change scores between baseline and 3 month were 0.44-0.67 and between baseline and 6 month were 0.46-0.48.Both the SRM and the ES scores of the JAFI dimensions were higher for the group without foot impairments after 3 month as compared to the group with persistent foot impairments . The JAFI impairment dimension showed the highest SRM and ES scores after ICI while the JAFI participation dimension showed the lowest scores. Correlations (rIn this study the JAFI appears to be a useful questionnaire for evaluating change in self-reported foot disability after treatment with ICI in children with JIA.None declared."} +{"text": "TLR10 is monomorphic, while the other nine TLR genes have between two and 12 alleles. 40 SNPs (16 non-synonymous) were identified across the ten TLR genes. These markers provide a springboard to future studies on innate immunity in the koala, a species under threat from two major infectious diseases.Toll-like receptors (TLRs) play a crucial role in the early defence against invading pathogens, yet our understanding of TLRs in marsupial immunity is limited. Here, we describe the characterisation of nine TLRs from a koala immune tissue transcriptome and one TLR from a draft sequence of the koala genome and the subsequent development of an assay to study genetic diversity in these genes. We surveyed genetic diversity in 20 koalas from New South Wales, Australia and showed that one gene, Phascolarctos cinereus) is an arboreal herbivorous marsupial which was widespread across eastern Australia until the end of the 19th century, when populations have suffered declines due to the fur trade, habitat degradation and disease [Environmental Protection and Biodiversity Conservation Act 1999. Genetic diversity in koalas is extremely low on the introduced islands and in Southern Australia due to founder effects [Trichosurus vulpecula) [The koala ,8.Chlamydia pecorum and C. pneumoniae [C. pecorum shedding in individual koalas with and without clinical disease [Many koala populations are currently threatened by habitat loss and vehicular injuries as well eumoniae , obligateumoniae . In wildeumoniae . However disease , would s disease studies disease . KoRV, o disease .Homo sapiens; TLR1\u201310), cow (Bos taurus) and pig (Sus domesticus), 12 in house mouse (Mus musculus) [Monodelphis domestica) [Sarcophilus harrisii) [Toll-like receptors (TLRs), encoded by a range of TLR genes, are key components of the innate immune response. TLRs are the first receptors to interact with invading microorganisms by recognising pathogen-associated molecular patterns (PAMPs) on a wide spectrum of pathogens . TLRs arusculus) \u201318 and 1mestica) and Tasmarrisii) . The TLRarrisii) , ssRNA (arrisii) and unmearrisii) . Non-virarrisii) ,25, lipoarrisii) , flagellarrisii) . Althougarrisii) . Interesarrisii) , raisingGenomics technologies have facilitated rapid elucidation of the basic architecture of the koala\u2019s immune system. Recent studies have described the characterisation of MHC class I and II ,8,30, inThe koala transcriptome dataset consists of four cDNA libraries which were established from immune related tissues including liver, lymph node, spleen and bone marrow . All tisHomo sapiens) and mouse (Mus musculus) TLR coding sequences. Koala TLR13 was obtained by searching the draft koala genome using BLAST using a Tasmanian devil (Sarcophilus harrisii) TLR13 nucleotide coding sequence [Gallus gallus), one amphibian and one fish species were analysed in MEGA 5 [Nine koala TLR genes were identified by searching all 4 koala cDNA libraries using BLsequence . Phylogesequence , one birn MEGA 5 using thn MEGA 5 with 100Genetically diverse koalas were selected in an attempt to maximize the TLR gene diversity discovered in this study. Control Region mitochondrial diversity was used as a proxy measure to choose 20 genetically diverse animals, representing six koala mitochondrial DNA (mtDNA) Control Region haplotypes . These koalas were all from New South Wales ranging from Narrandera (southern NSW) to Kyogle (northern NSW). Koala samples were either collected by the Port Macquarie Koala Hospital when animals were brought in for veterinary care or from the Australian Museum Tissue Collection. Australian Museum registration numbers are provided in 4)2SO4, 0.2 mM each dNTP, 2.0 mM MgSO4, 0.5 uM each forward and reverse primers, 1.5U of Platinum Taq DNA Polymerase High Fidelity and approximately 30 ng template DNA. The general cycle conditions were initial denaturation at 94\u00b0C for 2 min, followed by 35 cycles of 30 s denaturation at 94\u00b0C, 30 s annealing at 55\u201366\u00b0C and non-synonymous substitutions per non-synonymous sits (dN) in each gene were conducted using the Nei-Gojobori method with Jukes-Cantor adjustment in MEGA 5 [The overall rate of synonymous substitution per synonymous site . The evolutionary relationships of these genes are depicted in Based on the well characterized human TLR sequences, the koala TLR genes contained all of the key functionally conserved residues in the extracellular domain S2\u2013S11 FThe phylogenetic tree shown in The phylogenetic tree allows us to speculate about the evolutionary history of the TLR1 gene family. A single TLR1-like gene and a single TLR2 gene are seen in zebrafish and Xenopus . The chiThe TLR genes evolve independently in different lineages in response to species-specific pathogens . Genes iGenetic polymorphisms were detected in all koala TLR genes, except TLR10. In the nine polymorphic genes, the number of alleles per gene ranged from 2\u201312, and the number of SNPs per gene between 1\u20138 . In thisThe level of TLR diversity that was observed in koalas in this study is comparable to that observed in other species . For insS) and non-synonymous (dN) substitutions [S is higher than dN in all nine koala TLR genes characterized here. No non-synonymous substitutions were observed in TLR2 and TLR1/6-like Click here for additional data file.S2 FigBoxes and arrows means the locations of primers . Vertical lines show the boundaries of Leucine-Rich Repeats, transmembrane and cytoplasmic region.(TIF)Click here for additional data file.S3 Fig(TIF)Click here for additional data file.S4 Fig(TIF)Click here for additional data file.S5 Fig(TIF)Click here for additional data file.S6 Fig(TIF)Click here for additional data file.S7 Fig(TIF)Click here for additional data file.S8 Fig(TIF)Click here for additional data file.S9 Fig(TIF)Click here for additional data file.S10 Fig(TIF)Click here for additional data file.S11 Fig(TIF)Click here for additional data file.S12 FigAmino acid alignment of coding sequences of TLR2 in koala, Tasmanian devil, gray short-tailed opossum, house mouse, cattle and human. Dashes in the sequences represent gaps. Dots represent conservation of amino acids with the koala sequence. The ruler has been adjusted according to the koala TLR sequence. The LRR motifs are in green and are marked above with the consensus pattern: LxxLxLxxNxL according to human TLRs . The pos(TIF)Click here for additional data file.S13 FigThe stars above the residues are predicted pathogen binding positions in human TLR3 . The pre(TIF)Click here for additional data file.S14 FigThe predicted pathogen binding positions are in box according to human TLR4 .(TIF)Click here for additional data file.S15 FigThe predicted pathogen binding positions are in box according to human TLR5 .(TIF)Click here for additional data file.S16 FigThe predicted pathogen binding positions are in box according to human TLR6 .(TIF)Click here for additional data file.S17 FigThe predicted pathogen binding positions are in box according to human TLR7 .(TIF)Click here for additional data file.S18 FigThe predicted pathogen binding positions are in box according to human TLR8 .(TIF)Click here for additional data file.S19 FigThe predicted pathogen binding positions are in box according to human TLR9 .(TIF)Click here for additional data file.S20 FigThe predicted pathogen binding positions are in box according to human TLR10 .(TIF)Click here for additional data file.S21 Fig(TIF)Click here for additional data file.S1 TableXenopus tropicalis TLR9 that was from Ensemble.Amino acid sequence accession numbers were all obtained from GenBank, except (XLS)Click here for additional data file.S2 Table(XLS)Click here for additional data file.S3 Table(XLS)Click here for additional data file.S4 Table(XLS)Click here for additional data file."} +{"text": "Callithrix jacchus) as a preclinical nonhuman primate model to study reproductive and stem cell biology. The neonatal marmoset monkey ovary contains numerous primitive premeiotic germ cells (oogonia) expressing pluripotent stem cell markers including OCT4A (POU5F1). This is a peculiarity compared to neonatal human and rodent ovaries. Here, we aimed at culturing marmoset oogonia from neonatal ovaries. We established a culture system being stable for more than 20 passages and 5 months. Importantly, comparative transcriptome analysis of the cultured cells with neonatal ovary, embryonic stem cells, and fibroblasts revealed a lack of germ cell and pluripotency genes indicating the complete loss of oogonia upon initiation of the culture. From passage 4 onwards, however, the cultured cells produced large spherical, free-floating cells resembling oocyte-like cells (OLCs). OLCs strongly expressed several germ cell genes and may derive from the ovarian surface epithelium. In summary, our novel primate ovarian cell culture initially lacked detectable germ cells but then produced OLCs over a long period of time. This culture system may allow a deeper analysis of early phases of female primate germ cell development and\u2014after significant refinement\u2014possibly also the production of monkey oocytes.We use the common marmoset monkey ( The cultured ovarian cells were highly proliferative at low passages as revealed by Ki-67 staining. At higher passages (P9), OCCs showed a reduced number of Ki-67-positive cells. Highly proliferative ES cells were used as positive control. The primary antibody was used in a 1:300 dilution. The scale bar represents 50 \u03bcm.Supplementary Table 1. Ovary vs. OCCs top 50 up-regulated genes corresponding to data base identifier."} +{"text": "Coronary artery disease (CAD) accounts for half of all deaths in patients with end stage kidney disease (ESKD) who have undergone renal transplantation. In patients with ESKD, \"angiographically\" significant CAD may be asymptomatic. Identification of these patients, with subsequent revascularisation, may reduce the prevalence of adverse cardiovascular events in the peri-transplant and post-transplant period. We aimed to evaluate the effectiveness of diagnostic (>85% maximum age-predicted heart rate) dobutamine stress cardiac magnetic resonance (DSCMR) imaging in identifying \"angiographically\" significant CAD in asymptomatic patients.Over a five year period, 62 high-risk patients were referred to this program by the nephrology team. All of these subjects had an invasive coronary angiogram (ICA). These subjects had ESKD and were being considered for renal transplantation. 42 (68%) were male. All had at least one traditional cardiovascular risk factor. 58 (94%) were on renal replacement therapy. Of the 62 enrolled patients, 43 (69%) had a diagnostic DSCMR followed by an ICA. ICA reporters were blinded to results of DSCMR and vice-versa. 19 patients (31%) were excluded from the analysis due to non-diagnostic DSCMR scans. The most common reasons for a nondiagnostic DSCMR included insufficient augmentation in heart rate with dobutamine stress (in 8 patients) and claustrophobia (in 5 patients). Significant CAD was defined on ICA as a coronary stenosis of \u2265 70%.Of the 43 included patients, 12 (28%) had significant CAD, and all of these patients had evidence of inducible myocardial ischaemia on DSCMR. 3 (7.0%) patients had false positive DSCMR scans. There were no false negative scans. Of the 19 patients with non-diagnostic CMR scans, 7 patients had significant CAD on ICA. In this cohort studied with a diagnostic DSCMR: sensitivity = 100%, specificity = 90%, positive predictive value = 80%, negative predictive value = 100%. Over this period, 26 patients have undergone successful renal transplantation and 8 patients have died .When feasible, a diagnostic DSCMR can reliably detect \"angiographically\" significant CAD in patients with ESKD being considered for renal transplantation.Nothing to declare."} +{"text": "Global longitudinal strain and mitral annular plane systolic excursion (MAPSE) have been validated as an echocardiographic measure of longitudinal function in patients with aortic stenosis (AS). Cardiac magnetic resonance imaging (CMR) offers a simple and easy measure of MAPSE and been validated in various patient groups. Longitudinal function in aortic stenosis is usually reduced before a reduction in ejection fraction is seen. We sought to investigate the impact of TAVI upon longitudinal function and global ejection fraction in patients treated for symptomatic severe AS.We prospectively enrolled 52 patients with symptomatic severe aortic stenosis undergoing TAVI over a 5 year period from March 2009 to March 2014. Patients with contraindications to CMR were excluded and all patients provided informed written consent. All patients underwent an identical 1.5T CMR protocol at baseline and at a median of 6 months following TAVR. Multi-slice, multi-phase cine imaging was performed using a standard steady-state free procession pulse sequence in the short and long axis 340 mm) to cover the entire left and right ventricle. Longitudinal atrioventricular motion was measured at the septal and lateral mitral annulus in the 4 chamber cine view were studied. There was no significant change in the overall LV ejection fraction 6 months following TAVI compared to baseline. All three measures of longitudinal function were significantly improved 6 months following TAVI . Presence or absence of late gadolinium enhancement did not impact on the baseline MAPSEaverage or have impact on the change in MAPSEaverage following TAVI .52 patients male, AVA 0.6 \u00b1 0.2 cmLongitudinal left ventricular function, as assessed using MAPSE, improved following TAVI for severe aortic stenosis. MAPSE may be a more sensitive marker of left ventricular recovery following TAVI than global LV ejection fraction.This study was part funded by the British Heart Foundation (PG/11/126/29321)."} +{"text": "Neisseriameningitidis serogroup W strain circulating in England. The recent introduction of this vaccine into the United Kingdom national immunization program should also help protect infants against this endemic strain.Serum samples from children immunized with a meningococcal serogroup B vaccine demonstrated potent serum bactericidal antibody activity against the hypervirulent Neisseriameningitides (MenW) have been causal organisms for 1%\u20132% of IMD cases annually. An increase in invasive MenW disease associated with travel to the Hajj pilgrimage route during 2000\u20132002 was rapidly controlled after the introduction of mandatory vaccination for pilgrims has been declining in the United Kingdom since the early 2000s . Bexsero is composed of NHBA , NadA (Neisseria adhesin A) and fHbp (factor H binding protein), with meningococcal outer membrane vesicles from MenB strain from an outbreak in New Zealand has historically been recommended for high-risk persons and travelers to disease-endemic regions and for controlling outbreaks and fHbp variants (variant 2 peptide 22) among endemic MenW:cc11 isolates (http://pubmlst.org/perl/bigsdb/bigsdb.pl?db\u00a0=\u00a0pubmlst_neisseria_mrfgenomes) for nadA and nhba and, where present, their respective allelic and peptide variants.A total of 73 invasive MenW:cc11 isolates from England and Wales were received by the Public Health England Meningococcal Reference Unit during July 2010\u2013June 2013. These isolates were queried within the Meningitis Research Foundation Meningococcus Genome Library against 6 invasive MenW:cc11 isolates from patients 4 months\u201391 years of age in whom meningitis or septicemia was diagnosed in different regions of England and Wales during 2011\u20132012. SBA titers were expressed as the reciprocal of the serum dilution corresponding to nadA allele 5 (for peptide NadA-2/3.6) and nhba allele 17 (for NHBA peptide 29). Of the remaining 2 isolates, 1 isolate had a nadA allele that differed at a single nucleotide (C476A), causing a single amino acid change ; the other isolate had an nhba allele that differed at a single nucleotide (A376C), causing a single amino acid change .Of the 6 isolates tested, 4 possessed >1:32 against all 6 MenW isolates, independently of the immunization schedule . In adolescents, a single MenACWY dose would provide direct protection and, by targeting the age group most likely to carry meningococci . However, the effectiveness of Bexsero against meningococcal carriage and, therefore, its ability to provide herd protection, which is a major objective of an adolescent programme, is less certain than with conjugate vaccines (Conversely, infants would require"} +{"text": "We present comprehensive comparative evolutionary analysis of cGAS and OAS1 primate sequences and observe positive selection at nucleic acid binding interfaces and distributed throughout both genes. Our data revealed homologous regions with strong signatures of positive selection, suggesting common mechanisms employed by unknown pathogen encoded inhibitors and similar modes of evasion from antagonism. Our analysis of cGAS diversification also identified alternately spliced forms missing multiple sites under positive selection. Further analysis of selection on the OAS family in primates, which comprises OAS1, OAS2, OAS3 and OASL, suggests a hypothesis where gene duplications and domain fusion events result in paralogs that provide another means of escaping pathogen inhibitors. Together our comparative evolutionary analysis of cGAS and OAS provides new insights into distinct mechanisms by which key molecular sentinels of the innate immune system have adapted to circumvent viral-encoded inhibitors.A diverse subset of pattern recognition receptors (PRRs) detects pathogen-associated nucleic acids to initiate crucial innate immune responses in host organisms. Reflecting their importance for host defense, pathogens encode various countermeasures to evade or inhibit these immune effectors. PRRs directly engaged by pathogen inhibitors often evolve under recurrent bouts of positive selection that have been described as molecular \u2018arms races.\u2019 Cyclic GMP-AMP synthase (cGAS) was recently identified as a key PRR. Upon binding cytoplasmic double-stranded DNA (dsDNA) from various viruses, cGAS generates the small nucleotide secondary messenger cGAMP to signal activation of innate defenses. Here we report an evolutionary history of cGAS with recurrent positive selection in the primate lineage. Recent studies indicate a high degree of structural similarity between cGAS and 2\u2019-5\u2019- A pathogen\u2019s ability to infect new individuals within and across species is largely driven by its capacity to hijack cellular machinery and overcome the immune system. Pathogens have evolved multiple means to evade and shut down host immunity. Typically, mechanisms of inactivation involve direct interactions between host and pathogen factors. To escape inhibition over the course of generations, host factors frequently evolve in a manner that disrupts interactions at specific interfaces with pathogen factors. Likewise, pathogens adapt to restore such interactions, and these genetic tug-of-wars have been described as \u201cmolecular-arms races.\u201d Here we focus on the adaptation of two critical host immune factors, cGAS and OAS that share identity in protein structures despite very limited genetic similarity. Our analysis identifies a variety of ways, including amino acid changes on protein surfaces, by which these host factors appear to escape pathogen-mediated inhibition. Surprisingly, some amino acid substitutions are located at equivalent sites suggesting that cGAS and OAS may have adapted to evade common pathogen encoded inhibitors. These data also identify protein surfaces that are targeted by viruses to inhibit host immunity. Taken together our results indicate the existence of critical, yet-to-be identified viral antagonists of cGAS and OAS. The bpattern recognition receptors (PRRs), initiate immune responses upon recognition of pathogen macromolecular structures (Reviewed in [oligoadenylate synthase (OAS) family of proteins [A set of host genes, termed iewed in ,5). Becaiewed in ,6. PRRs iewed in ,5. Multiiewed in ,5. Two oproteins and the proteins which approteins \u201311. Becaproteins ,12,13, wn) (where n > = 2 and <20) upon RNA binding [nucleotidyltransferase (NTase) within the ClassI-CCase family and OAS1-C terminal domain [OAS proteins are cytoplasmic dsRNA binding proteins that generate the second messenger 2\u2019-5\u2019 oligoadenylate [in vitro [cGAS provides complementary surveillance as a cytoplasmic double-stranded DNA binding protein that app (STING) ,29\u201331, w (STING) ,29. cGAS (STING) ,32,33 an (STING) ,35, whicin vitro . cGAS hain vitro ,37 and ein vitro .Vibrio cholerae [The initial characterization of cGAS highlighted several parallels with OAS mediated defenses : 1) nuclcholerae , which acholerae ,41, suggHere, we focus on more recent evolution of cGAS and OAS to compare how these nucleic acid sensors have been influenced by selection from pathogens. Consistent with their vital role in immune surveillance ,13,39, wCyclic GMP-AMP synthase (cGAS), previously referred to as C6ORF150, provides a primary block against viruses ,38 and idN/dS). The sites model implemented in Phylogenetic Analysis by Maximum Likelihood (PAML) [dN/dS values per amino acid position and compares models that omit or accommodate elevated dN/dS to test for positive selection. Our alignment of primate cGAS orthologs revealed signatures of positive selection (PARtitioning approach for Robust Inference of Selection (PARRIS) algorithm from the HyPhy package [To determine if cGAS evolved under positive selection in primates, we cloned and sequenced cDNA of cGAS from 22 simian primates calculat<0.0001) . We furt package , which a package . PARRIS dN/dS values at each branch in our primate phylogeny using the free-ratio model in PAML. Consistent with a critical role as a host defense gene antagonized by specific viral inhibitors, cGAS exhibits dN/dS ratios exceeding one\u2014a hallmark of positive selection\u2014on various branches in hominoid, Old World, and New World monkey lineages : 1 synonymous (S) amino acid changes). We carried out complementary analysis of episodic selection using the GA-Branch and aBSREL test in HyPhy [To investigate whether cGAS has been subject to episodic positive selection during primate evolution, we calculated lineages . The brain HyPhy binding of viral nucleic acids, 2) generation of small nucleotide secondary messengers containing 2\u2019-5\u2019 phosphodiester bonds, and 3) use of these secondary messengers to activate an antiviral response ,39. In adN/dS values, similar to cGAS , consist to cGAS . We obse to cGAS supportidN/dS ratios compared to our analysis of cGAS. The complementary MEME, and FUBAR tests (HyPhy package) identified multiple residues overlapping with PAML analysis under positive selection in OASSimilar to cGAS, multiple amino acid positions are under selection in OAS1 . PhylogedN/dS mapped to protein surfaces of cGAS contact dN/dS values on matching branches of the primate lineage. This analysis uncovered evidence of a surprising correlation , while OAS3 was near the significance cut-off are well-documented for contributions to transcript diversity and regulation . AlternaWe recovered several alternatively spliced cDNAs of cGAS in hominoid, Old World, and New World Monkey species , consistThe Red Queen hypothesis provides a useful framework for investigating recurrent genetic conflicts like those unfolding at host-pathogen interfaces . To dateOAS proteins are encoded by an ancient and dynamic gene family characterized by extensive duplications in some mammalian lineages ,16,17. IIntriguingly, only a few sites appear under positive selection in OAS3 with even the more sensitive methods of detection , despiteAs another potentially adaptive mechanism we identified multiple primate cGAS isoforms that encode intact ORFs. Intriguingly we found four isoforms that cleanly excise all of exon 3 from cGAS, which contains three sites under positive selection. Importantly, spliceforms that lack exon 3 but maintain exon 2 still contain the cGAMP catalytic residues. Based on published cGAS domain deletion data and the Regardless of mechanism, alternative splicing has been noted in several cases for evasion of pathogens. Alternative splicing of human APOBEC3G, 3F, and 3H has been documented with varying impacts on antiviral activity and susceptibility to Vif antagonism ,60. SuppConsistent with their critical role as PRRs ,64, our in vitro and in vivo [dN/dS ratios for these sites (posterior probability >0.99) suggest that this domain may be a prime target for pathogen inhibitors of cGAS.The localization of amino acid positions under positive selection can identify new interfaces involved in protein-protein interactions between host and pathogen factors . Notably in vivo . However in vivo , the stadN/dS values) for matching branches in the primate tree using BLAT searches. For other primates, sequences were obtained by Sanger sequencing of PCR amplicons using cDNA as a template or genomic DNA. Briefly, cDNA was synthesized using Superscript III mastermix (Life Technologies) or Maxima cDNA synthesis kit (Thermo) from total RNA extracted from fibroblast cell lines obtained from Coriell. Sequences of interest were PCR amplified from cDNA using Phusion High-Fidelity mastermix (Thermo) according to the manufacturer\u2019s instructions and analyzed by 1\u20132% agarose gel electrophoresis. Amplicons of interest were excised, purified using Zymo gel extraction kit, and subject to Sanger sequencing or TOPO cloned (Life Technologies) followed by sequencing. For cGAS sequences from New World Monkeys, each exon was PCR amplified from genomic DNA. DNA sequences were analyzed using Geneious software.DNA sequence alignments were carried out using MUSCLE with default settings in Geneious. All sequences are available in DNA sequences were manually trimmed to remove indels and aligned using Geneious v6.1.7 (Biomatters Ltd.) using default settings. This alignment and a species trees representing currently accepted primate relationships were useWe carried out permutation tests by generating two vectors representing cGAS and OAS1 of length 40 to represent 40 amino acids of the helical spine. Executing 1,000,000 trials we determined the probability of getting three sites overlapping between the two vectors (the R script is included in http://www.cgl.ucsf.edu/chimera/)[Amino acids identified as being under positive selection using PAML and Datamonkey were mapped onto the three-dimensional crystal structures of the apoform of cGAS (PDB: 4KM5) and DNA chimera/).Total RNA from primate fibroblast cell lines treated with 1000 U of interferon/mL was extracted using the RNAeasy kit (Qiagen). 1\u20132 \u03bcg of total RNA was reverse-transcribed using the Maxima cDNA synthesis kit (Thermo). cDNA was diluted to a final volume of 50 \u03bcL of which 1 \u03bcl was used as a template for PCR. PCR was carried using Phusion according to the manufacturer\u2019s protocol for 35 cycles using cGAS Fint 5\u2019-accgggagctactatgagca-3\u2019 and cGAS Rint 5\u2019-tgtcctgaggcactgaagaa-3\u2019primers. PCR amplicons were analyzed using 2% agarose gel electrophoresis.S1 FigA phylogenetic tree produced by the PhyML plugin in Geneious using 22 primate cGAS cDNA sequences.(EPS)Click here for additional data file.S2 FigLineages identified by GA-Branch as being subject to positive selection are labeled in red.(EPS)Click here for additional data file.S3 FigIn instances where aBSREL was unable to calculate a value (S = 0), the number of nonsynonymous changes relative to synonymous changes calculated by PAML free-ratio analysis are shown. Lineages displaying \u03c9 > 1 or at least 3 nonsynonymous changes are highlighted in red.(EPS)Click here for additional data file.S4 Fig(A) The co-crystal of the mouse cGAS dimer with dsDNA (PDB:406A) with sites under positive selection (red). (B and C) Two interfaces where cGAS sites under positive selection interact with DNA. Each cGAS monomer is individually colored either blue or turquoise. (D) Amino acid alignment of primate variation for the two cGAS rapidly evolving sites that contact DNA in the mouse dimer crystal structure.(EPS)Click here for additional data file.S5 FigA phylogenetic tree produced by the PhyML plugin in Geneious using 22 primate cGAS cDNA sequences.(EPS)Click here for additional data file.S6 FigLineages identified by GA-Branch as being subject to positive selection are labeled in red.(EPS)Click here for additional data file.S7 Fig(A) The co-crystal of OAS1 (yellow) and dsRNA (silver) with sites under positive selection labeled in red. (B and C) Two OAS1 protein surfaces that interact with dsRNA are highlighted. (D) Amino acid alignment showing primate variation for OAS1 at rapidly evolving sites that contact RNA in the crystal structure.(EPS)Click here for additional data file.S8 FigdN/dS, specifically those with dS > 0.01.Models of sequence evolution were used to estimate the rate of each protein on each branch of our primate tree. Plotting each branch by its rate in OAS1 and cGAS reveals a clear linear correlation, indicating that rapid evolution in one gene is typically paralleled by rapid evolution in the other gene. Rates were estimated in the free-ratio branch model of PAML. The correlation analysis was restricted to branches with sufficient divergence to provide reliable estimates of (EPS)Click here for additional data file.S9 Fig. In instances where aBSREL was unable to calculate a value (S = 0), the number of nonsynonymous changes relative to synonymous changes calculated by PAML free-ratio analysis are shown. Lineages displaying \u03c9 > 1 or at least 3 nonsynonymous changes are highlighted in red.(EPS)Click here for additional data file.S10 Fig(A) cGAS RT-PCR splicing assay. (B) cGAS spliceform RT-PCR amplicons resolved by 2% agarose gel electrophoresis. Numbering of spliceforms the same as in (EPS)Click here for additional data file.S11 FigcGAS spliceform variant predicted sequences are high(EPS)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.S4 Table(DOCX)Click here for additional data file.S5 Table(DOCX)Click here for additional data file.S6 Table(DOCX)Click here for additional data file.S7 Table(XLSX)Click here for additional data file.S8 Table(DOCX)Click here for additional data file.S9 Table(DOCX)Click here for additional data file.S10 Table(DOCX)Click here for additional data file.S11 Table(DOCX)Click here for additional data file.S12 Table(DOCX)Click here for additional data file.S13 Table(DOCX)Click here for additional data file.S1 Dataset(TXT)Click here for additional data file."} +{"text": "Trafficking and expansion of myeloid derived suppressor cells (MDSCs) plays a major role in immune suppression of tumor. MDSCs express chemokine receptors which likely mediate their recruitment to the tumor microenvironment. Suppression of T cells is also mediated by the interaction of programmed death-1 (PD-1) and its ligands which are abundantly expressed in cancer cells and immune infiltrates, including MDSCs. Here, we show that targeting both pathways through administration of a small molecule chemokine receptor antagonist (CCX9588 which blocks CCR1) and an anti-PDL1 antibody significantly reduced tumor burden in an orthotopic breast cancer mouse model. Primary tumor growth and lung metastasis were modestly reduced by either agent (anti-PDL-1 or CCR1 inhibitor) alone, but the combination of CCR1 inhibitor plus anti-PDL1 antibody resulted in significantly reduced tumor burden as compared to either of the single agents. Orthotropic breast cancer cell engraftment induces robust expansion of CD11b+Ly6Ghi Ly6Chi MDSCs, a subpopulation of which express CCR1. Analysis of the tumor infiltrating cells revealed that CCX9588 significantly reduced the number of MDSCs in primary tumors, suggesting that CCR1 blockade of MDSC trafficking translates into reduced tumor burden. Finally, analysis of human breast cancer patient samples from The Cancer Genome Atlas (TCGA) database revealed that the CCR1 ligands, CCL3 (MIP-1a) and CCL5 (RANTES) are present at significantly higher levels in breast cancers as compared to normal breast tissue. These data are the first to show that CCR1 chemokine receptor antagonists can act synergistically with PDL-1 inhibitors, and suggest a novel approach for potentiating the activity of immune cell checkpoint inhibitors."} +{"text": "The somatic embryogenesis tissue culture process has been utilized to propagate high yielding oil palm. Due to the low callogenesis and embryogenesis rates, molecular studies were initiated to identify genes regulating the process, and their expression levels are usually quantified using reverse transcription quantitative real-time PCR (RT-qPCR). With the recent release of oil palm genome sequences, it is crucial to establish a proper strategy for gene analysis using RT-qPCR. Selection of the most suitable reference genes should be performed for accurate quantification of gene expression levels.pOP-EA01332, PD00380 and PD00569) outperformed classical housekeeping genes . PD00380 and PD00569 were identified as the most stably expressed genes in total samples, MA2 and MA8 tissue culture lines. Their applicability to validate the expression profiles of a putative ethylene-responsive transcription factor 3-like gene demonstrated the importance of using the geometric mean of two genes for normalization.In this study, eight candidate reference genes selected from cDNA microarray study and literature review were evaluated comprehensively across 26 tissue culture samples using RT-qPCR. These samples were collected from two tissue culture lines and media treatments, which consisted of leaf explants cultures, callus and embryoids from consecutive developmental stages. Three statistical algorithms confirmed that the expression stability of novel reference genes (PD00380 and PD00569 were selected for accurate and reliable normalization of gene expression data from RT-qPCR. These data will be valuable to the research associated with the tissue culture process. Also, the method described here will facilitate the selection of appropriate reference genes in other oil palm tissues and in the expression profiling of genes relating to yield, biotic and abiotic stresses.Systematic selection of the most stably expressed reference genes for RT-qPCR was established in oil palm tissue culture samples. Elaeis guineensis), which originated from West Africa, is a diploid monocotyledon that belongs to the Arecaceae family and Elaeidinae sub-tribe Oil palm , EgER6 (Ethylene responsive 6), Eg707 (unknown protein) and EgIAA9 (putative member of the AUX/IAA gene family) However, the challenge encountered by tissue culturists is the low efficiency of the process itself. The callogenesis rate of leaf explants is around 19% Accurate quantification of gene expression levels using RT-qPCR is highly dependent on the normalization of GOI with the most suitable reference genes. Recent developments have shown that more than one reference gene is required for optimum normalization of non-biological sample-to-sample variation introduced during RT-qPCR ACTINGAPDH) GRAS), cyclophilin 2 (CYP2) and pre-mRNA splicing factor 7 (SLU7) as reference genes in the study of oil palm leaf discs subjected to various abiotic stresses Similar to other plant species, RT-qPCR studies in oil palm utilized the classical housekeeping genes such as PD00380), manganese superoxide dismutase (PD00569), predicted protein IFH-1 like (pOP-EA01332), GAPDH, NADH dehydrogenase subunit 5-like gene (NAD5), alpha-tubulin 1 (TUBULIN), polyubiquitin (UBIQUITIN) and actin (ACTIN)] were investigated in this study across samples collected from various consecutive developmental stages of oil palm tissue culture, with cultured leaf explants sampled at different days, callus and embryoids (EMB). Different tissue culture lines and media treatments were also used. Detailed and systematic analyses were carried out using geNorm PD00380 and PD00569) were selected as the most stably expressed genes compared to classical housekeeping genes. Application of these genes to normalize the expression levels of an ethylene-responsive transcription factor 3-like gene (PD00088) in oil palm is also discussed.Given the significance of the tissue culture process to the oil palm industry, expression stability of eight candidate reference genes suggested in preliminary studies by Low PD00380 (EY397675), PD00569 (EL682210) and pOP-EA01332 (EY406625), which were identified from an oil palm cDNA microarray study across embryogenic callus (EC), non-embryogenic callus (NEC), EMB, shoot from polyembryoids (ST), female inflorescence (INF), kernel at 12 weeks after anthesis (WAA), mesocarp at 15 WAA and roots from six months old seedling palms GAPDH (DQ267444), NAD5 (DQ872924), TUBULIN (EL685625), UBIQUITIN (EL689143) and ACTIN (AY550991), which were selected based on a literature review A total of eight candidate genes were selected for determination of the most stable reference genes across various developmental stages of oil palm tissue culture. These samples which constituted leaf explant cultures, callus and EMB from consecutive developmental stages were collected from two different tissue culture lines and media treatments . The selected genes were novel reference genes or classical housekeeping genes. As shown in pOP-EA01332, which was only associated to three GO terms under the biological process. The majority of the genes are involved in cellular and metabolic processes, in cell or organelle components and engaged in binding or catalytic activities values for most of the genes were greater than 0.99, which signified a strong correlation between the cycle threshold (Ct) values and the amount of cDNA template used in the amplification reactions.For each of the candidate reference genes, a standard curve was generated across each tissue culture line with different media treatments . The estNAD5 and UBIQUITIN, with mean Ct values in the range of 17 to 24 cycles. The pOP-EA01332 gene exhibited the lowest expression levels compared to other candidate reference genes. Preliminary statistical analysis using the coefficient of variation (CV) was performed to determine the most stably expressed gene across this set of samples GAPDH, NAD5 and TUBULIN exhibited higher variation in their gene expression levels. The CV values for these genes were from 4.76 to 8.91 of reference genes. In the event the value of V is lower than the recommended cutoff value of 0.15, addition of expression data from another reference gene is not required for calculation of a normalization factor based on the geometric mean 2+3 for each set of data were less than 0.15. Therefore, only two reference genes, PD00380 and PD00569 are needed for accurate normalization of all the datasets in this study.In addition to ranking the genes according to ACTIN, PD00380 and PD00569 were the top three most stably expressed genes across all the tissue culture samples, MA2 and MA8 datasets followed by GAPDH (3.91\u00b10.83) exhibited least stable expression levels. Grouping of reference genes according to expression stability were consistent with the output generated from geNorm and NormFinder. A distinct expression stability cluster was detected between novel and classical reference genes. The former was always grouped as the most stable cluster while the latter formed the least stable cluster.BestKeeper is an Excel based spreadsheet software application .95\u00b10.80 . SimilarPD00380 and PD00569 as the most significantly correlated genes in total samples across the tested samples and Week_3 (W3) leaf explants from MA2 and MA8 tissue culture lines that were cultured on media T527 and T694. PD00088 encodes for a putative ethylene-responsive transcription factor 3-like gene that contains a binding site for the AP2 DNA binding domain, which is postulated to be involved in somatic embryogenesis. A previous study using a cDNA microarray platform showed that the transcript for PD00088 was highly expressed in W1 and W3 leaf explants from media T527 as compared to media T694 across MA2 and MA8 tissue culture lines oil palm reference genes and five classical oil palm housekeeping genes were chosen for evaluation across tissue culture samples. Primer pairs for these genes were carefully designed to ensure amplification of specific PCR products from reverse transcribed cDNA. The presence of a single amplicon peak in the melting curve analysis as the best combination of reference genes in the longan system MnSOD) was classified as the least stable gene in longan SE as opposed to oil palm. Studies in Nicotiana plumbaginifolia revealed that the expression of MnSOD was induced in the plant cells during conditions of metabolic stress in tissue culture The oil palm SE process in this study was initiated from leaf explants, which dedifferentiated into callus and subsequently somatic embryos. The presence of tissue culture samples from undifferentiated and differentiated phases across two auxin concentrations have made the selection of reference genes quite challenging as distinct groups of genes will be expressed. In order to increase the chances of selecting the most suitable reference genes, classical housekeeping genes were evaluated in parallel with novel reference genes. This approach also proved useful for longan SE and resulted in the recommendation of Medicago truncatulaHevea brasiliensisMtSERF1 from M. truncatula was detected in the embryogenic calli and globular somatic embryo, while several AP2/ERF genes from Hevea were highly expressed in the calli from the embryogenic line. Oil palm PD00088 (a putative ethylene-responsive transcription factor 3-like) belongs to this superfamily. Due to its potential substantive role in SE, this gene was selected to validate the applicability of PD00380 and PD00569 as reference genes. Results from RT-qPCR , tenera fruit type \u00d7100%PCR amplification efficiencies, Ex \u200a=\u200a[10(slope represents the slope of linear regression)This was followed by the RT-qPCR of these primers across individual cDNA samples (10 ng) from MA2T527, MA2T694, MA8T527 and MA8T694.The Ct values for each sample were retrieved using Realplex software version 2.2 . Data analysis was carried out in Microsoft Excel. Average Ct values from three replicates were calculated and transformed into relative expression quantities using the delta Ct method, Ex\u2227(minCt - sampleCt). The most stable reference genes across the tissue culture samples were selected based on geNorm v3.4 File S1List of top 75 gene clones identified from analysis of cDNA microarray datasets from tissue culture materials and mature tissues.(XLS)Click here for additional data file.File S2Determination of the most stably expressed genes across tissue culture materials and mature tissues using geNorm software. Expression levels for each reference gene were measured across tissue culture materials and mature tissues . Average expression stability values (M) was calculated for each reference gene. The least stable genes with higher M values were excluded in a stepwise manner until the most stable reference genes were shortlisted.(DOC)Click here for additional data file.Figure S1Determination of PCR amplification efficiencies (Ex) and correlation coefficient (2R) values for PD00569 using the slope of standard curve. The estimated Ex for PD00569 ranged from 88 to 104% and the 2R were given as 0.9926 to 0.9989.(DOC)Click here for additional data file.Figure S2Determination of the most stably expressed reference genes across media treatment T527 and T694 using geNorm software. Average expression stability values (M) were calculated for each reference gene. The least stable genes with higher M values were excluded in a stepwise manner until the two most stable reference genes were obtained for the tested tissue culture media.(DOC)Click here for additional data file.Figure S3Melting curve generated for PD00569 across tissue culture samples collected from MA2 and MA8 tissue culture lines. The presence of a single amplicon peak indicated the amplification of a specific PCR product.(DOC)Click here for additional data file.Table S1Preliminary statistical analysis of oil palm candidate reference genes using the coefficient of variation (CV).(DOC)Click here for additional data file.Table S2Pair-wise correlation analysis and correlation analysis of oil palm candidate reference genes across the total samples (MA2 and MA8 tissue culture lines).(DOC)Click here for additional data file.Table S3Pair-wise correlation analysis and correlation analysis of oil palm candidate reference genes across the MA2 tissue culture line.(DOC)Click here for additional data file.Table S4Pair-wise correlation analysis and correlation analysis of oil palm candidate reference genes across the MA8 tissue culture line.(DOC)Click here for additional data file."} +{"text": "Oxidative stress occurs with disturbed blood flow, inflammation and cardiovascular disease (CVD), yet free-radical scavenging antioxidants have shown limited benefit in human CVD. Thioredoxin-1 (Trx1) is a thiol antioxidant protecting against non-radical oxidants by controlling protein thiol/disulfide status; Trx1 translocates from cytoplasm to cell nuclei due to stress signaling, facilitates DNA binding of transcription factors, e.g., NF-\u03baB, and potentiates inflammatory signaling. Whether increased nuclear Trx1 contributes to proatherogenic signaling is unknown.In vitro and in vivo atherogenic models were used to test for nuclear translocation of Trx1 and associated proinflammatory signaling. Disturbed flow by oscillatory shear stress stimulated Trx1 nuclear translocation in endothelial cells. Elevation of nuclear Trx1 in endothelial cells and transgenic (Tg) mice potentiated disturbed flow-stimulated proinflammatory signaling including NF-\u03baB activation and increased expression of cell adhesion molecules and cytokines. Tg mice with increased nuclear Trx1 had increased carotid wall thickening due to disturbed flow but no significant differences in serum lipids or weight gain compared to wild type mice. Redox proteomics data of carotid arteries showed that disturbed flow stimulated protein thiol oxidation, and oxidation was higher in Tg mice than wild type mice.Translocation of Trx1 from cytoplasm to cell nuclei plays an important role in disturbed flow-stimulated proatherogenesis with greater cytoplasmic protein oxidation and an enhanced nuclear transcription factor activity. The results suggest that pharmacologic interventions to inhibit nuclear translocation of Trx1 may provide a new approach to prevent inflammatory diseases or progression. Blood flow generates shear stress on vascular endothelial cells (EC) and regulates endothelial biology and cardiovascular disease (CVD), including atherosclerosis. Atherosclerotic lesions develop at arterial bifurcations and branch points that are exposed to patterns of disturbed or oscillatory blood flow, whereas straight arterial segments exposed to uni-directional, laminar flow are atherosclerosis-resistant 2O2, NO, UV, viral infection, and cadmium Thioredoxin (Trx) containing a pair of redox-active cysteine (Cys) in its catalytic site is the major cellular oxidoreductase enzyme regulating cellular redox homeostasis 62) in the DNA binding region of p50 NF-\u03baB and inhibits DNA binding 38 of p65 NF-\u03baB plays a crucial role in DNA binding 2O2-dependent transcriptional termination mechanism.Compartmental regulation of NF-\u03baB, AP-1 and Nrf-2 involves opposing redox-sensitive steps in cytoplasm and nuclei, i.e., upstream cytoplasmic oxidative activation involves kinase signaling, and downstream stimulation of DNA binding activity occurs through Trx1-dependent reduction of Cys in the DNA-binding domain Modulation of NF-\u03baB signaling by nuclear Trx1 raises the possibility that excessive nuclear Trx1 could cause a proinflammatory response accompanying hyper-responsive immune signaling. Indeed, our previous study using a transgenic mouse model in which nuclear Trx1 was increased due to expression of a fusion protein containing human Trx1 with a nuclear localization signal (NLS) showed that NF-\u03baB activity is enhanced by nuclear Trx1 In the present study, we examined the role of nuclear Trx1 in atherogenic signaling using mouse and cultured EC models. The results from an atherogenic mouse model show that increased Trx1 in cell nuclei stimulated disturbed flow-induced carotid wall thickening, activated NF-\u03baB and elevated VCAM level with no significant increase in mouse weight gain or blood lipid levels. Proteins in the carotid artery were substantially oxidized by disturbed flow and this oxidation was enhanced by nuclear overexpression of Trx1. Transient overexpression of Trx1 in cell nuclei of EC potentiated OS-dependent NF-\u03baB activation and elevated proinflammatory cytokine and cell adhesion molecule genes. Together, the data show that increased nuclear Trx1 causes disruption of nuclear function that potentiates atherogenic signaling and represents a potential target for therapeutic intervention.This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the Emory University. All protocols involving mice in this study were reviewed and approved by the Emory University Institutional Animal Care and Use Committee (IACUC).2 as described previously in vitro, cells were transiently transfected with NLS-Trx1 in nuclei Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza , cultured in M199 media with 20% fetal bovine serum and used between passages 5 and 6 on 0.1% gelatin-coated 100-mm dishes at 37\u00b0C and 5% COTo examine Trx1 nuclear translocation by shear stress, subcellular fractionation of endothelial cells exposed to shear stress was performed using Qproteome kit following the procedures provided by the manufacturer. Isolated fractions were then confirmed by Western blotting probed with \u03b2-actin and lamin for cytoplasm and nuclei antibodies, respectively. Trx1 levels in these fractions were determined by Western blotting probed with Trx1 antibody . Alexa-Fluor-680-conjugated anti-rabbit or anti-mouse secondary antibody (Invitrogen) was used and a band corresponding to each protein was visualized using an Odyssey scanner and Odyssey 2.1 software .Total RNA was polyadenylated and reverse transcribed for use in a two-step qRT-PCR using the High-capacity cDNA Synthesis kit (ABI) and qRT-PCR kits (Stratagene) as described d-flow) in LCA and stable flow (s-flow) in RCA was confirmed at one day post-ligation by Doppler ultrasonography using the Vevo770 system with a 30-MHz probe (RMV707B) Transgenic male mice (8 weeks of age) expressing human Trx1 in cell nuclei , with eThe present study shows that disturbed oscillatory shear stress induces translocation of Trx1 into vascular endothelial cell nuclei and that increased nuclear Trx1 potentiates proinflammatory signaling by NF-\u03baB and atherogenesis in a mouse model of CVD. Earlier studies show that Trx redox state is controlled differently in subcellular compartments. The mitochondria are the most reduced compartment and mitochondrial Trx2 is highly sensitive to oxidation in vivo studies with this well-established partial carotid ligation model for CVD clearly show an important contribution of increased nuclear Trx1 to the known pathophysiologic pathway.The present studies show that redox regulation of NF-\u03baB activation due to increased nuclear Trx1 has an important role in potentiating vascular pathophysiology in the mouse partial carotid ligation model of atherogenesis. The results with the NLS-Trx1 Tg mouse showed that NF-\u03baB activity was increased, transcripts for cell adhesion molecules were increased, and significant carotid artery wall thickening and increased Oil-Red-O staining were evident at the region exposed to disturbed blood flow. In these studies with increased nuclear Trx1, no detectable changes in serum lipid levels or weight gain was observed, indicating that the proatherogenic effects observed in NLS-Trx1 Tg mouse were not due to indirect effects on lipid metabolism. In comparative studies (not shown), we also examined effects of overexpression of Trx1 in cytoplasm using a transgenic mouse line expressing Trx1 fusion protein containing a nuclear export signal and Trx2 in mitochondria using a transgenic mouse line expressing human Trx2 and observed no stimulation of inflammation or proatherogenic signaling. Thus, the in vivo results were confirmed in vitro; overexpression of Trx1 in HUVEC nuclei showed potentiation of NF-\u03baB activation and elevation of cell adhesion molecule and cytokine by OS. In addition to NF-\u03baB regulation, elevated Trx1 in nuclei resulted in increased mRNA expression of Bax and procaspase-3 but decreased Bcl-2 (data not shown). These genes are closely associated with activation of cell death and regulated by HIF1\u03b1 and AP-1 transcription factors The in vitro data also show differential responses of EC to LS and OS. However, it is still largely unknown how OS and LS induce signaling differently ultimately resulting in opposite cellular responses, atheroprotection and atheropromotion. Therefore, further studies with systematic approaches are required, for instance, identification of the source for oxidant generation because redox signaling is involved in both LS and OS responses. This study will be needed to provide information for Trx1 nuclear translocation mechanism since previous studies Shear stress is mechanotransduced into a biochemical signal that results in changes in vascular behavior. Maintenance of steady laminar blood flow is crucial for normal healthy vascular function while disturbed oscillatory flow near arterial bifurcations and curvatures is associated with atherosclerosis. In addition, vascular endothelial cells respond differently to different shear flow at the molecular and cellular levels, and such responses are associated with opposite consequences either preventing or promoting atherosclerosis. In the present study, 2O2-stimulated endothelial cell death was prevented by direct interaction of Trx1 with actin 73 residue by acrolein, resulted in stimulation of monocyte adhesion, an early step in atherogenesis Shear stress regulates endothelial cell alignment and structure by controlling actin cytoskeletal reorganization As shown in our previous study In summary, the current study shows that increased Trx1 in cell nuclei potentiates atherogenesis. The results show that oscillatory shear induced nuclear translocation of Trx1 and that increased Trx1 caused enhanced carotid wall thickening, lipid accumulation and proinflammatory signaling in a disturbed flow model of CVD. Redox proteomics showed that disturbed flow resulted in oxidation of actin cytoskeleton proteins in mouse carotid artery and this oxidation was elevated by increased nuclear Trx1. Taken together, the data show that nuclear translocation of Trx1 is a critical step of endothelial dysfunction induced by disturbed oscillatory flow. Because cytoplasmic oxidative stress and other stress signals stimulate nuclear translocation of Trx1 in different cell types, the results suggest that excessive nuclear Trx1 due to diverse stressors may be a general contributor to the prevalence of CVD. Consequently, the results suggest new therapeutic targets to modulate Trx1 nuclear translocation and compartmental redox systems to prevent or delay atherosclerosis.Table S1Redox ICAT/MS-measured % oxidation of peptidyl Cys/protein of RCA and LCA in WT mice. The data show 146 peptidyl Cys with % oxidation (%ox) value identified from RCA and LCA of WT mice.(PDF)Click here for additional data file.Table S2Redox ICAT/MS-measured % oxidation of peptidyl Cys/proteins of RCA and LCA in NLS-Trx1 Tg mice. The data show 193 peptidyl Cys with % oxidation (%ox) value identified from RCA and LCA of Tg mice.(PDF)Click here for additional data file.Table S3Redox ICAT/MS-measured % oxidation of peptidyl Cys/proteins of LCA in WT and Tg mice. The data show 156 peptidyl Cys with % oxidation (%ox) value identified from LCA of WT and Tg mice.(PDF)Click here for additional data file."} +{"text": "We reported a case of patent ductus arteriosus (PDA) with congenital anomaly of coronary arteries as abnormal origin of right coronary artery (RCA) and left coronary artery (LCA) from a single ostium of the right coronary sinus. A 21-year-old man referred to our institution for evaluation of cardiac murmur. He has suffered from palpitation and atypical chest pain for three months. On physical examination, a continuous murmur was heard in the second left parasternal space. Transthoracic echocardiography showed normal left and right ventricular size and systolic function (LVEF = 55%). Main pulmonary artery (PA) and left pulmonary artery (LPA) branch were considerably dilated. Considering normal coronary flow, lack of clinical evidence of myocardial ischemia and echocardiography findings, patient underwent surgical closure of PDA via left thoracotomy and after five days discharged uneventfully. We reported a case of patent ductus arteriosus (PDA) with congenital anomaly of coronary arteries as abnormal origin of right coronary artery (RCA) and left coronary artery (LCA) from a single ostium of the right coronary sinus. 2 saturation study showed normal RV pressure and pulmonary artery pressure. Aortic root injection depicted that both RCA and LCA originate from the right coronary sinus through a single ostium, LAD was located superiorly with a benign anterior course . Main pulmonary artery (PA) and left pulmonary artery (LPA) branch were considerably dilated. A small (5-6mm) PDA with insignificant left to right shunt was detected without r course . AortogrIn an anatomic collection of congenital heart disease by Frescura et al. , an isolated anomalous origin of coronary arteries was reported to be 2.2% , both right and left coronary artery from the right sinus were reported to be 15% (4 of the 27 patients) . In revi"} +{"text": "They smoked deniconized (denic) and average nicotine (avnic) cigarettes during the PET scans.Twenty healthy American male smokers who abstained from tobacco overnight were genotyped and completed positron emission tomography (PET) scans with the mu opioid receptor agonist, [ND) of [11C]carfentanil in the right medial prefrontal cortex , left anterior medial prefrontal cortex , right ventral striatum , left insula , right hippocampus and left cerebellum , and increased the BPND in left amygdala , left putamen and left nucleus accumbens . In the AA allele carriers, avnic cigarette smoking significantly changed the BPND compared to after denic smoking in most brain areas listed above. However in the *G carriers the significant BPND changes were confirmed in only amPfc and vStr. Free mu opioid receptor availability was significantly less in the *G than the AA carriers in the Amy and NAcc.Smoking avnic cigarette decreased the binding potential (BPND changes induced by avnic smoking in OPRM1 *G carriers were blunted compared to the AA carriers. Also *G smokers had less free mu opioid receptor availability in Amy and NAcc.The present study demonstrates BP Many studies with mice have demonstrated that nicotine induces endogenous brain opioid release . FurtherAdditional basic science studies support the importance of the opioid system, especially the mu opioid receptor (OPRM1), in drug addiction. Humanized h/mOPRM1-118 AA or h/mOPRM1-118 GG receptors (knockin) mice show different reinforcement of alcohol. The GG mice have a four-fold greater vStr/NAcc DA release to alcohol than the former . Also th11C]carfentanil binding potentials (MOR BPND) compared to AA carriers with 0.6 mg of nicotine (nic) cigarette smoking. Reduced BPND assumes increased endogenous opioid release and less free mu opioid receptors (activation). They also found that *G carriers had a positive association between decreased MOR BPND and smoking reward. The present study reports the role of OPRM1 A118G in brain endogenous opioid release following tobacco smoking as measured by [11C]carfentanil displacement with denic and 1.0 mg nicotine avnic cigarettes.11C]raclopride by . Significant decreases in BPND after avnic compared to after denic were found in the ventral striatum (vStr), left insula (Ins), right hippocampus (Hippo), left anterior medial prefrontal cortex (amPfc), left medial prefrontal cortex (mPfc), and left cerebellar vermis in the amygdala (Amy), nucleus accumbens (NAcc), and putamen (Put) in the left hemisphere , left Ins , right Hippo , left amPfc , left mPfc , left Cbl , Amy and NAcc . In the *G carriers, the BPNDs were significantly different between cigarette smoking conditions in vStr and left amPfc .The subjects were genotyped and separated into two groups based on the existence OPRM1 G allele. There were 15 AA and 5 *G (GA and GG) carriers. For each genotype, BPThere were fewer free mu opioid receptors in the left Ins , vStr and right Hippo and NAcc for human PET studies with [11C]carfentanil (11C]carfentanil is less than 15 % of the therapeutic dose for conscious sedation (ND was only measured in this study. The BPND after denic smoking was considered the baseline to compensate for non-specific binding (which was unlikely to occur). Before tobacco smoking MOR BPND in abstinent smokers and nonsmokers should be studied.The present study has major limitations: (1) Limited number of subjects. (2) Lack of controls for the tobacco smokers. (3) Females were not studied (due to a lack of approved research funds for the role of sex differences). The number of G allele carriers was also limited. Larger groups including tobacco smokers and nonsmokers, males as well as females must be studied for Effect size and Power to be >0.8. (4) Since [fentanil . The dossedation and is oIn conclusion this small genetic/PET study indicates that the OPRM1 A118G genotype altered tobacco smoking/nicotine effects on endogenous brain opioid release. The AA allele smokers had more free mu opioid receptor availability."} +{"text": "Synthetic folic acid has been shown to cause serious, potentially life-threatening IgE mediated anaphylaxis -3. SynthA search of the Canada Vigilance Database from April 1, 2008 to August 31, 2013 identified 15 cases of anaphylaxis and 16 reports of serious non-anaphylactic allergic reactions.All 15 cases of anaphylaxis involved multi ingredient products with NHP doses (up to and including 1 mg). Fourteen cases were female (93%). One case of anaphylaxis and 3 cases of serious allergic reaction involved women taking prenatal supplements, two of which reported spontaneous abortion. There was insufficient case information available to assess any causal association between the allergic reaction and the fetal loss. Three medically important cases involved pharmaceutical doses of folic acid (5 mg), one of which was a pediatric case involving a multi ingredient intravenous total parental nutrition product (Multi-12/K1 pediatric) containing folic acid.Confirmatory allergy tests for folic acid were not available for any of the cases. None of the reports noted a known diagnosis of folic acid allergy.Folic acid allergy appears to be rare and would not be expected to be well known among health providers or consumers. Increasing awareness amongst health providers regarding folic acid allergy may improve the identification and counselling of patients with this allergy."} +{"text": "We analyzed signs and symptoms in 90 patients diagnosed with acute HIV infection in a community-based program that offered universal HIV-1 nucleic acid amplification testing. Forty-seven (52%) patients reported ongoing signs or symptoms at the time of testing. Another 25 (28%) reported signs or symptoms that had occurred during the 14 days before testing. The detection of acute HIV infection (AHI) is critical to HIV prevention and treatment strategies has been provided to all rapid antibody\u2013negative participants since June 2007 (samples for NAT are obtained at the time of rapid antibody testing) (>2 signs/symptoms) was defined according to criteria described by Braun et al. , we obtained blood samples for CD4 and viral load testing and collected detailed information regarding occurrence, duration, and start and stop dates for 11 signs and symptoms associated with AHI were excluded. The University of California San Diego\u2019s Human Research Protections Program approved the study protocol, consent process, and all study-related procedures.For statistical analysis, SPSS version 21 was used. For analysis on signs or symptoms compatible with AHI, signs or symptoms that started All 90 participants were male and self-identified as men who have sex with men (MSM). Median age was 29 (range 18\u201367) years. Half (50%) of participants reported white race; 29% reported Hispanic ethnicity. Median number of male partners reported for the previous 12 months was 20 (IQR 14\u201331). A total of 72 (80%) patients had signs or symptoms associated with AHI that occurred within 2 weeks before undergoing NAT; of these 72 patients, 47 (52% of the study population) had ongoing signs or symptoms, while signs or symptoms had resolved by the time of testing for 25 (28% of the study population). Twelve (13%) reported signs or symptoms starting after testing, while 6 (7%) reported the absence of signs or symptoms . A totalOverall, 69 patients (77%) reported signs or symptoms that met criteria of compatibility with AHI . Onset oData on whether a patient sought medical attention because of signs or symptoms were available for 42 (47%) of 90 patients; of these, 12 (29%) reported that they sought medical attention because of their signs or symptoms and 30 (71%) did not. Significantly higher viral loads were observed for those who sought medical attention compared with those who did not .Overall, 70 (78%) of the 90 patients fulfilled criteria for having typical AHI and 20 (22%) did not . Patients with typical AHI had significantly higher viral loads compared with patients without (p<0.01). A total of 61 (85%) of 72 patients with signs or symptoms before NAT testing fulfilled criteria for having typical AHI. In addition, 40 (85%) of 47 patients who had ongoing signs or symptoms at the time of AHI testing fulfilled criteria for having typical AHI at that time.We characterized signs or symptoms relative to the date of AHI diagnosis among persons seeking HIV testing in a program offering universal AHI screening. Two findings are notable: 1) 52% of participants reported ongoing signs or symptoms at the time of AHI testing, and 2) 80% reported signs or symptoms occurring within 2 weeks before undergoing testing.These findings may have major clinical implications for community-based settings that restrict AHI testing to persons with ongoing signs or symptoms. This practice may be relatively insensitive in settings where MSM undergo HIV screening frequently . Estimates on frequency of signs and symptoms in HIV-negative persons ranged widely in previous studies. Although in one study a specificity of 65% was estimated for influenza illness\u2013like symptoms may increase the yield of AHI diagnoses by more than half."} +{"text": "Proliferative thyroid lesions including nodular goiter and hyperplastic lesion are very common in the Middle East and North African (MENA) region . HyperplWhole transcript expression profiles were generated in 17 goiters, 14 hyperplastic lesions and 7 TN samples utilizing Affymetrix HuGene 1.0 ST arrays. We used the default analysis method for generating a threshold of significance for differential expression . Partek Genomics Suite and Ingenuity Pathway Analysis software packages were utilized to interpret data sets.Expression profiles of goiters and hyperplastic lesions were highly related and no transcripts were differentially expressed between these two thyroid lesions under the given statistical threshold values. However, more than 10000 genes were differentially expressed between goiters, as well as hyperplastic lesions, and TN samples. The most differentially expressed transcripts were in fact downregulated in both thyroid lesions in comparison to TN samples and include genes like olfactory receptor, family 6, subfamily N (OR6N2), glial cells missing homolog 1, Drosophila (GCM1), family with sequence similarity 138, member B (FAM138B), prostate-specific P704P mRNA (P704P), and olfactory receptor, family 5, subfamily H, member 14 (OR5H14). The most upregulated transcripts in goiters and hyperplastic lesions vs TN samples include genes as cytochrome c oxidase assembly protein COX15 homolog (COX15), dyskeratosis congenita 1, dyskerin (DKC1), and DnaJ (Hsp40) homolog, subfamily A, member 2 (DNAJA2). Networks which were most deregulated in goiter and hyperplastic lesion in comparison to TN tissue share similar functions although certain pathways seem to be differentially affected in both thyroid lesions.Our study indicates that goiter and hyperplastic lesion share common deregulated expression profiles in comparison to TN tissue. As a certain number of goiters and hyperplastic lesions bear the capacity to develop to thyroid neoplasms, knowledge of deregulated genes in these lesions may help to identify patients which are at elevated risk for developing thyroid carcinoma. Further studies have to reveal which expression signatures in these benign thyroid lesions are in common with malignant cases.This study was supported by King Abdulaziz City for Science and Technology (KACST) grants 09-BIO707-03 and 09-BIO820-03."} +{"text": "Leishmania spp. are protozoan parasites that have two principal life cycle stages: the motile promastigote forms that live in the alimentary tract of the sandfly and the amastigote forms, which are adapted to survive and replicate in the harsh conditions of the phagolysosome of mammalian macrophages. Here, we used Illumina sequencing of poly-A selected RNA to characterise and compare the transcriptomes of L. mexicana promastigotes, axenic amastigotes and intracellular amastigotes. These data allowed the production of the first transcriptome evidence-based annotation of gene models for this species, including genome-wide mapping of trans-splice sites and poly-A addition sites. The revised genome annotation encompassed 9,169 protein-coding genes including 936 novel genes as well as modifications to previously existing gene models. Comparative analysis of gene expression across promastigote and amastigote forms revealed that 3,832 genes are differentially expressed between promastigotes and intracellular amastigotes. A large proportion of genes that were downregulated during differentiation to amastigotes were associated with the function of the motile flagellum. In contrast, those genes that were upregulated included cell surface proteins, transporters, peptidases and many uncharacterized genes, including 293 of the 936 novel genes. Genome-wide distribution analysis of the differentially expressed genes revealed that the tetraploid chromosome 30 is highly enriched for genes that were upregulated in amastigotes, providing the first evidence of a link between this whole chromosome duplication event and adaptation to the vertebrate host in this group. Peptide evidence for 42 proteins encoded by novel transcripts supports the idea of an as yet uncharacterised set of small proteins in Leishmania spp. with possible implications for host-pathogen interactions. Leishmania are single-celled parasites that are transmitted between animal hosts by the bite of sand flies. Once inside their animal hosts they abandon their extracellular habit and invade cells of the immune system, called macrophages. This oscillation between hosts requires the parasite to be able to adapt to dramatically different environments. To help unravel the multitude of biochemical, ultrastructural and lifestyle differences that distinguish these specialised life cycle stages we characterised and contrasted the transcriptomes of insect and mammalian adapted forms. Using bioinformatic approaches we revised the genome annotation and discovered nearly 1,000 new genes that had not been described before. We found that over 3,000 genes change in their expression to facilitate the change in host environment including those involved in specifying cell shape, extracellular appearance and biochemistry. Furthermore we reveal that an ancient chromosome duplication shared by all Leishmania species may have contributed to the adaptation of these globally important parasites to parasitism of vertebrates. Trypanosoma brucei, T. cruzi and Leishmania spp. affect mostly people in developing countries and together account for 4.4M disability-adjusted life years (XLSX)Click here for additional data file.S2 TableGFF feature record Columns are , , , , , (\u201c.\u201d denotes no score), (\u201c.\u201d denotes not relevant), , [attribute](XLSX)Click here for additional data file.S3 TableGFF feature record Columns are , , , , , (\u201c.\u201d denotes no score), (\u201c.\u201d denotes not relevant), , [attribute](XLS)Click here for additional data file.S4 TableGFF feature recordColumns are , , , , , (\u201c.\u201d denotes no score), (\u201c.\u201d denotes not relevant), , [attribute](XLS)Click here for additional data file.S5 TableGFF feature record Columns are , , , , , (\u201c.\u201d denotes no score), (\u201c.\u201d denotes not relevant), , [attribute] Every novel transcripts was given a unique IDs in the format LmxM.[number of chromosome]_[position of last nucleotide of stop codon of predicted CDS], for example: LmxM.01_107651.(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S7 Table(XLSX)Click here for additional data file.S8 Table(XLSX)Click here for additional data file.S9 Table(XLSX)Click here for additional data file.S10 Table(XLSX)Click here for additional data file.S11 TableGFF feature record Columns are , , , , , (\u201c.\u201d denotes no score), (\u201c.\u201d denotes not relevant), , [attribute](XLS)Click here for additional data file.S12 Table(XLSX)Click here for additional data file.S13 Table(XLSX)Click here for additional data file.S14 Table(XLSX)Click here for additional data file.S15 Table(XLSX)Click here for additional data file.S16 Table(XLSX)Click here for additional data file.S17 Table(XLSX)Click here for additional data file.S18 Table(DOCX)Click here for additional data file.S19 Table(XLSX)Click here for additional data file.S20 Table(XLSX)Click here for additional data file.S21 Table(XLSX)Click here for additional data file.S22 Table(XLSX)Click here for additional data file."} +{"text": "Gait has been generally viewed as a largely automated motor task, requiring minimal higher-level cognitive input. Increasing evidence, however, suggest that attention demanding cognitive tasks to disturb gait,2. Movem21 athletes (age 25.57\u00b14.77 years) with clinically diagnosed FAI were recruited. All participants completed 5 trials of normal walking and 5 trials of normal walking while performing a cognitive task. The cognitive task consisted of subtracting seven from a randomly selected number between 11 and 99 repeatedly whilst walking. Three dimensional rotations of the affected ankle (measured by an eight-camera motion capture system at 100 Hz) were calculated by visual3D during gait cycles. Between trials variability of ankle rotations time curves during stance phase and during 200ms before and after heel strike were calculated using the coefficient of multiple correlations (CMC) and intraclass correlation (ICC)The results indicate that mean CMC was decreased during dual task condition in the sagittal and frontal planes. This was statistically significant in frontal plane during 200ms before and after heel strike (p<0.05) Table . There wThe athletes with FAI demonstrated greater ankle movement variability during dual task condition which may indicate diminished neuromotor control. Cognitive load may increase episodes of ankle instability in these athletes.Nester declares a personal commercial interest in the insoles tested in this study."} +{"text": "Crush syndrome is often encountered in natural disasters. It is a critical condition leading to multiple organ failure. However, the mechanisms by which the local traumatic injuries affect distant organs remain unknown. We paid attention to bone marrow-derived mononuclear cells (BMMNCs) as therapeutic strategy against crush injury. Transplantation of BMMNCs can elicit protection and regeneration against damaged organs through their paracrine properties and its clinical application has been realized to treat ischemia reperfusion-related various diseases. We have previously reported that multiple organ damage based on systemic inflammatory response is induced in crush injury and the pathogenesis is largely dependent on massive ischemia-reperfusion. We investigated whether BMMNCs could suppress systemic inflammation and improve mortality in a rat model of crush injury.7 cells in 1 ml phosphate-buffered saline (PBS)) immediately after weight removal (BMMNC group) and PBS alone was administered in the control group (CR group). The sham group underwent the same procedure without compression of hind limbs (SH group). The rats were observed over 7 days after the injury to evaluate survival. To estimate antiinflammatory effects of BMMNCs, sera were collected 3 hours, 6 hours and 24 hours after the injury. The levels of interleukin 6 (IL-6) and tumor necrosis factor alpha (TNFa) were measured by ELISA and statistically analyzed.To develop crush syndrome, both hind limbs of rats were compressed for 6 hours under weights (3.0 kg each). Rats in the treated group were intravenously administrated BMMNCs (1 \u00d7 10P < 0.05). Crush injury-induced upregulation of serum IL-6 was significantly reduced by the BMMNC treatment at all time points (P < 0.05). The level of TNFa decreased significantly in the BMMNC group compared with that in the CR group 24 hours after the compression release (P < 0.05). These findings suggest that transplantation of BMMNCs has an ability to evade the devastating condition following crush injury by suppressing systemic inflammation.The survival rate at day 7 in the BMMNC group was 80.0%, and that in the CR group was 47.6%, revealing that the 7-day survival was significantly improved by the BMMNC injection (The administration of BMMNCs reduced production of inflammatory cytokines and improved survival rate in a rat model of crush injury. Cell therapy using BMMNCs might become a novel therapy against crush injury."} +{"text": "There has been an increase in the use of non-invasive ventilation (NIV) in the intensive care setting. Guidelines suggest more favourable outcomes when used in patients with chronic obstructive pulmonary disease (COPD) and cardiogenic pulmonary oedema (CPO), yet itsTo review the practice and outcomes of NIV in our institution over a 1 year period.st January 2014 and 31st December 2014. Patients were divided into 3 groups: decompensated respiratory failure (DRF) (pH< 7.35 + pCO2>6KPa pre-NIV), non-decompensated respiratory failure (NDRF) (pH>7.35 or PCO2< 6Kpa pre-NIV) or post extubation. For each patient, the indication and duration of NIV was recorded along with important outcomes that included requirement and duration of mechanical ventilation (MV), length of ICU stay and survival to ICU discharge.We conducted a retrospective analysis of all patients receiving NIV for the first time at any stage of their ICU admission between 1103 patients received NIV as a first line respiratory supportive therapy for a number of indications of which only 20% included COPD or CPO , median duration of NIV received was similar (23 [IQR 11-105] and 17 [IQR 8-73.5] hours for DRF and NDRF respectively (p = 0.13)). There were no significant differences between DRF and NDRF with respect to % of patients requiring subsequent mechanical ventilation , median duration of mechanical ventilation for those subsequently intubated or % surviving to ICU discharge . There was a non-significant trend towards reduction in ICU length of stay (LOS) for those with DRF compared with NDRF .NIV following a period of mechanical ventilation was given for 31 patients. In this cohort, NIV was administered for a median duration of 21 hours (range 1-617 hours). 13/31 patients (42%) required re-intubation and further periods of mechanical ventilation.In our institution, the majority of patients received NIV for indications outside those recommended by guidelines. Those with DRF receiving NIV fared no worse compared with NDRF in this cohort. This along with those receiving NIV following extubation may provide a cohort of patients who warrant further investigation."} +{"text": "The origins of neointimal smooth muscle cells that arise following vascular injury remains controversial. Studies have suggested that these cells may arise from previously differentiated medial vascular smooth muscle cells, resident stem cells or blood born progenitors. In the current study we examined the contribution of the previously differentiated vascular smooth muscle cells to the neointima that forms following carotid artery ligation.Myh11) or smooth muscle \u03b1-actin (Acta2) driven tamoxifen regulated cre recombinase. Following treatment of adult mice with tamoxifen these mice express mEGFP exclusively in differentiated smooth muscle cells. Subsequently vascular injury was induced in the mice by carotid artery ligation and the contribution of mEGFP positive cells to the neointima determined.We utilized transgenic mice harboring a cre recombinase-dependent reporter gene (mTmG). These mice express membrane targeted tandem dimer Tomato (mTomato) prior to cre-mediated excision and membrane targeted EGFP (mEGFP) following excision. The mTmG mice were crossed with transgenic mice expressing either smooth muscle myosin heavy chain .Analysis of the cellular composition of the neointima that forms following injury revealed that mEGFP positive cells derived from either These data demonstrate that the majority of the neointima that forms following carotid ligation is derived from previously differentiated medial vascular smooth muscle cells. Vascular smooth muscle cells (VSMCs) are the major contractile components of the vascular system. They are critically important for regulating blood pressure and flow throughout the vascular system. Unlike skeletal and cardiac muscle cells, VSMCs are remarkably plastic and modulate their phenotype in response to extracellular cues during the development and progression of a variety of diseases including atherosclerosis, hypertension, stenosis following injury and restenosis following vascular interventions. Cardiovascular disease is the leading cause of death among the US population, yet despite intense research efforts a number of basic questions regarding the etiology of cardiovascular disease remain elusive. Classically these diseases are described as being associated with dedifferentiated VSMCs that have decreased expression of proteins required for the normal contractile function, increased expression of extracellular matrix proteins and increased cell proliferation. These pMyh11) creER(T2)-/+ mice[Acta2) creER(T2)-/+ mice[tm4Luo/J) . Slides were mounted in Prolong Gold containing DAPI (Invitrogen) and visualized by confocal microscopy (Olympus Fluoview FV1000). For immuno-fluorescent staining cryosections were permeabilized in 0.2% triton in TBS for 5\u00a0minutes, washed in TBS for 5\u00a0minutes them blocked in 5% goat serum diluted in TBS at room temperature for 1\u00a0hour. Blocked sections were incubated for 4-5 hours at 37\u00b0 with primary antibodies to, the SM2 isoform of smooth muscle myosin heavy chain (1:500), CD31 . Cells were collected by brief centrifugation , washed in the same buffer and recentrifuged. Pelleted cells were resuspended in phosphate buffered saline containing 0.5% bovine serum albumen, 0.5\u00a0mM EDTA and subjected to FACS analysis. Red blood cell fragments were excluded based on side scatter analysis and the remaining cells sorted based on their red and green fluorescence. A minimum of 10,000 cells were counted in each sample. For each analysis, cells obtained from a mouse that expresses EGFP ubiquitously (CAG-EGFP: C57BL/6-Tg(CAG-EGFP)131Osb/LeySopJ) were used as a positive control for EGFP-expressing cells. Cells obtained from a cre negative mTmG mouse were used as a positive control for mTomato expressing cells and cells obtained from a nontransgenic mouse were used as negative controls. Gates were established based on the distribution of the control cell groups and the numbers of cells in the experimental mice that fall into each group were determined.For FACS blood was harvested from mice immediately prior to tissue collection. Blood collected into heparinized tubes was diluted 10 fold into red blood cell lysis buffer Luo/J) in which all cells express a membrane localized mTomato tandem dimer in the absence of cre recombinase activity. Following cre-mediated excision of the mTomato cassette, cells express membrane localized EGFP[Myh11[Acta2[Myh11 or Acta2.Results of a recent study suggest that neointimal cells that form following vascular injury are derived from a stem cell population resident within the vascular wall rather than from previously differentiated VSMCs. In thatzed EGFP -/+ mTmG-/+ double transgenic mice[Acta2 creER(T2)-/+ mTmG-/+[Myh11 creER(T2)-/+ mTmG-/+ double transgenic mice we observed a significant decrease in expression of the endogenous Myh11 gene within the medial layer of carotid arteries as evidenced by decreased smooth muscle myosin isoform, SM2, immunostaining medial VSMCs. Immunostaining with antibodies to CD31 demonstrate that there are also a number of CD31 positive, mTomato positive endothelial like cells within the neointima -/+ mTmG-/+ double transgenic mice 28\u00a0days following carotid ligation the majority of neointimal cells also expressed mEGFP medial VSMCs it is possible that these cells may be mobilized from distant sites rather than being derived from locally resident medial VSMCs. To evaluate the existence of a population of mobile mEGFP positive cells we performed a fluorescence activated cell sorting (FACS) analysis of blood obtained from tamoxifen treated Myh11 creER(T2)-/+ mTmG-/+ double transgenic mice, 7,14 and 28\u00a0days following injury transgenic mice to show that following femoral artery wire injury the neointima that formed contained LacZ positive cells[Our data are consistent with and extend previous fate mapping studies using a cre-dependent LacZ reporter. In thisve cells. Togetheve cells. In furtAlthough our studies do not rule out the possibility that under appropriate, in vitro, culture conditions the expansion of a progenitor cell population may be favored, the current studies provide compelling evidence that, in vivo, the majority of neointimal cells that arise following carotid ligation are derived from differentiated medial VSMCs. The lack of detectable mEGFP positive cells circulating in the blood, further suggests that the neointimal cells likely arise from the dedifferentiation and migration of locally derived medial VSMCs.The authors declare that they have no competing interests.myh11 creER(T2) transgenic mice, aided in data interpretation and reviewed and edited the final manuscript. All authors reviewed and approved the final manuscript.BPH prepared the manuscript and figures, harvested tissues, obtained cryosections, performed all confocal microscopy, immunohistology and quantitative analysis. AMH maintained mouse lines, performed carotid ligations and harvested tissues. CB provided access and training in the use of the confocal microscope and performed FACS studies. SO generated the"} +{"text": "In managing women with large uterine fibroids, GnRH agonists can be used to reduce fibroid volume and decrease vascularity to enhance treatment outcomes with MRgFUS. GnRH agonists have been shown to reduce fibroid volume by as much as 30-40%. The purpose of this study was to examine the responsiveness of fibroids to pre-treatment with a GnRH agonist in relation to their appearance on T2 weighted images and MRgFUS treatment outcomes.Fifteen women (ages 34-52) with symptomatic uterine fibroids were pre-treated with a GnRH agonist prior to undergoing MRgFUS treatment using the ExAblate device. 7 patients received treatment for 3 months, 4 received treatment for 4 months, 2 received treatment for 6 months and 2 received treatment for an unknown time before the MRgFUS procedure. Other than the 2 patients with unknown GnRH agonist therapy times, all patients obtained their last GnRH agonist injection 3-4 weeks before MRgFUS treatment. These women were selected to receive a GnRH agonist if they possessed fibroids in excess of 10 cm or had a fibroid volume greater than 300 cc. Fibroids were classified by their intensity on T2 weighted images relative to normal myometrium . A total of 22 fibroids were treated with MRgFUS: 17 hypointense, 3 heterogeneously hypointense, 1 heterogeneously hyperintense and 1 isointense fibroid. Data regarding fibroid volume reduction following GnRH administration, Joules of energy delivered per cc of fibroid tissue ablated and the final non-perfused volume (NPV) were investigated.The average reduction in size of fibroids following GnRH treatment based on T2 appearance was: hypointense (22%), isointense (44%) heterogeneously hypointense (33%) and heterogeneously hyperintense (40%). The average Joules of energy delivered per sonication was greater for isointense (4550 J) and heterogeneously hyperintense (4200 J) fibroids than for heterogeneously hypointense (3227 J) and hypointense (2654 J) fibroids. Additionally, the volume of tissue ablation per Joule of energy applied was significantly larger for the heterogeneously hypointense (0.066 cm3) and heterogeneously hyperintense (0.057 cm3) than for the hypointense (0.036 cm3) and isointense (0.018 cm3). The NPV per fibroid was greater for heterogeneously hyperintense (85%), heterogeneously hypointense (63%) and hypointense (72%) fibroids than for isointense (32%) fibroids. Fibroid image characteristics may be used to predict the effectiveness of GnRH agonist therapy prior to MRgFUS treatment. While more vascular fibroids require greater energy to treat, they show a more favorable response to pre-treatment with a GnRH agonist in terms of fibroid volume reduction and thermoablative treatment effectiveness than hypointense fibroids."} +{"text": "Animal studies have reported that treatment with angiotensin II receptor blockers reduced kidney sodium-dependent glucose cotransporter expression. We therefore hypothesized that patients with hypertension treated with an angiotensin II receptor blocker (candesartan) would probably have an increased response to sodium-dependent glucose cotransporter inhibitor therapy (ipragliflozin) compared with patients treated with alternative hypertensive medications such as calcium channel blockers (nifedipine).Although sodium-dependent glucose cotransporter inhibitor (ipragliflozin) is a new anti-diabetic medicine, the clinical efficacy in the Japanese population has not been fully evaluated. We compared the combined effect of angiotensin II receptor blocker candesartan plus ipragliflozin with nifedipine plus ipragliflozin therapy and found that the combination of candesartan plus ipragliflozin was more effective in increasing glycosuria and lowering plasma glucose.P<0.05).A 57-year-old Japanese man with essential hypertension was treated with candesartan. Candesartan was switched to nifedipine for the initial 10 days of an observation period and 5 days later he was started on ipragliflozin (day 6 of nifedipine treatment) with nifedipine for the next 5 days. Thereafter (from day 11 to day 20), candesartan was started instead of nifedipine and ipragliflozin was continued. In the last 5 days ipragliflozin was stopped and he was treated with candesartan alone. Neither nifedipine alone (0.038+/-0.004) nor candesartan alone (0.048+/-0.006) produce any trace amount of glycosuria. However, the extent of glycosuria under ipragliflozin with candesartan treatment (37.5+/-8.45) was significantly greater than that of ipragliflozin with nifedipine (23.75+/-0.35; Candesartan demonstrated additive actions with ipragliflozin to increase glycosuria compared to ipragliflozin with nifedipine treatment. Sodium-dependent glucose cotransporter 2 (SGLT2) inhibitors are a new class of anti-diabetic agents and are currently attracting clinicians\u2019 attention in our country, Japan. HoweverA 57-year-old Japanese man with essential hypertension diagnosed 2 years earlier was treated with candesartan (8mg/day) following the initial diagnosis. He did not have any complications including diabetes mellitus except for essential hypertension based on annual comprehensive medical check-up. After he gave informed consent, candesartan (8mg/day) was switched to nifedipine 10mg/day) for the initial 10 days of an observation period .In Figure\u00a0This report describes a very important clinical observation that patients treated with an ARB (candesartan) will probably have an increased response to SGLT2 inhibitor therapy (ipragliflozin) compared with patients treated with alternative hypertensive medications such as CCBs (nifedipine). Although CCBs are not known to modulate SGLT2 function, ARB was recently reported to downregulate SGLT2 expression in a hypertension rat model. Thus, cThis case report also indicates another important aspect of SGLT2 therapy. SGLT2 inhibition is reported to reduce the threshold of glycosuria from 180mg/dL plasma glucose to 21mg/dL plasma glucose. TherefoBased on our observation, we speculated that patients treated with an ARB (candesartan) would probably have an increased response to SGLT2 inhibitor therapy (ipragliflozin) compared with patients treated with alternative hypertensive medications such as CCBs (nifedipine). Care will be required to carefully monitor patients treated with both ARBs and SGLT2 inhibitors to determine the extent of drug\u2013drug interactions to increase glycosuria and dehydration particularly in the aging patient population.Written informed consent was obtained from the patient for publication of this case report and any accompany images. A copy of the written consent is available for review by the Editor-in-Chief of this journal."} +{"text": "Cicer accessions. High intra- and inter-specific polymorphic potential and wider molecular diversity (11\u201394%) along with a broader genetic base (13\u201378%) specifically in the functional genic regions of wild accessions was assayed by mapped markers. It suggested their utility in monitoring introgression and transferring target trait-specific genomic (gene) regions from wild to cultivated gene pool for the genetic enhancement. Distinct species/gene pool-wise differentiation, admixed domestication pattern, and differential genome-wide recombination and LD estimates/decay observed in a six structured population of wild and cultivated accessions using mapped markers further signifies their usefulness in chickpea genetics, genomics and breeding.Characterization of natural allelic diversity and understanding the genetic structure and linkage disequilibrium (LD) pattern in wild germplasm accessions by large-scale genotyping of informative microsatellite and single nucleotide polymorphism (SNP) markers is requisite to facilitate chickpea genetic improvement. Large-scale validation and high-throughput genotyping of genome-wide physically mapped 478 genic and genomic microsatellite markers and 380 transcription factor gene-derived SNP markers using gel-based assay, fluorescent dye-labelled automated fragment analyser and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass array have been performed. Outcome revealed their high genotyping success rate (97.5%) and existence of a high level of natural allelic diversity among 94 wild and cultivated Cicer arietinum (L.)] is one of the most important food legume crops in the world and serves as an important source of protein with essential amino acids in human diet. To improve the productivity and sustainability of agriculture, the development of high-yielding durable stress tolerant (climate resilient) cultivars is the prime objective in current chickpea genomics and molecular breeding research. The systematic and extensive breeding efforts of chickpea have resulted in the development of a number of high-yielding stress tolerant cultivars through intra- and inter-specific hybridization of a few selected germplasm accessions. Therefore, the genetic base of modern chickpea cultivars is narrow which in turn limits the yield enhancement and increased stress tolerance. Furthermore, the earlier archaeological, phenotypic and molecular phylogeny studies inferred that chickpea domestication might have led through maximum adaptation-based selection pressure, followed by punctuation by a series of four sequential evolutionary bottlenecks during their evolutionary transition from wild C. reticulatum, which progressively narrows down the genetic base in cultivated gene pool Cicer includes many annual and perennial wild species that represent a valuable and potential source of diverse gene pool for improving yield and abiotic/biotic stress tolerance in cultivated chickpea Chickpea for each 384 SNP loci were designed using MassARRAY Assay Design software v3.1. All other default parameters except for the optimal amplicon size containing the SNP sites of 80\u2013120 bp were set in the software for analysis. The EP and UEP were optimized for 34-plex assays and procured oligos from IDT, USA. A 10-mer tag (5\u2032-ACGTTGGATG-3\u2032) was added to the 5\u2032-end of each forward and reverse pre-amplification primers to improve the PCR performance and mass spectrometry. The amplification of synthesized pre-amplification primers with multiplex PCR assay, shrimp alkaline phosphate (SAP) incubation for neutralizing unincorporated dNTPs, primer extension, resin cleanup and mass spectrometry were performed following the manufacturer's instructions of Sequenom iPLEX Gold amplification kit (http://www.sequenom.com). The SNP genotyping results obtained in iPLEX spectrochip bio-arrays were analyzed using MassARRAY Typer 3.4 software tool. The different SNP alleles among 94 accessions were visualized and documented based on allele-specific differences in mass between extension products.These TF gene-derived 384 SNPs were further genotyped in the genomic DNA of 94 cultivated and wild Cicer species The genotyping information of validated 478 microsatellite markers and 380 TF gene-derived SNP markers was analysed in PowerMarker v3.51 Cicer accessions. The cluster analysis among accessions was performed based on Nei's genetic distance The genotyping data of 858 markers (including 478 microsatellite and 380 SNP markers) physically mapped across eight chromosomes of chickpea were used to determine functional molecular diversity and population structure and for establishing genetic relationships among 94 cultivated and wild Cicer accessions were used in a model based program STRUCTURE v2.3 ad hoc method of Pritchard et al. Cicer accessions was constructed. Various population genetic parameters including the efficiency of markers for estimating genetic divergence (FST) and degree of admixture among different population groups was estimated. Analysis of molecular variance (AMOVA) among accessions and populations, and within populations and accessions was performed using both GenALEX 6.4 The genome-wide 858 marker genotyping data generated from 94 wild and cultivated 2 (average correlation coefficient) among 94 cultivated and wild Cicer accessions belonging to diverse population groups (as inferred by population structure). The significance of LD estimates was determined at 100,000 permutations. The marker loci mapped on the same and different chromosomes were considered as linked and unlinked markers, respectively. The LD was estimated using the genotyping information of linked, unlinked and global (both linked and unlinked) markers. To determine the LD occurrence due to physical linkage of marker loci, 99th percentile of r2 distribution for unlinked markers was considered as background level of LD 2 values of marker-pairs located in the physical intervals of 0\u2013200, 200\u2013400, 400\u2013600, 600\u2013800, 800\u20131000 and 1000\u20131200 kb across eight chromosomes of chickpea. The graph was plotted between pooled r2 and physical distance (kb) based on nonlinear regression model considering the r2 value \u200a=\u200a1 at marker physical distance of 0 kb Cicer accessions. The statistical significance test comparing r2 values of LD across population groups was performed using ANOVA interface tool of SPSS v17.0 .The correlation in frequency among pairs of alleles across a pair of 858 marker loci that are physically mapped on eight chromosomes were analysed using TASSEL v2.1 Cicer accessions using the agarose gel-based assay and fluorescent dye-labelled automated fragment analyser . Four hundred seventy-eight (including 334 genic and 144 genomic microsatellite markers) of 496 microsatellite markers (96%) showed polymorphism among 94 Cicer accessions . Moreover, we discovered 384 novel TF gene-derived high-quality SNPs showing differentiation between ICC 4958 and ICC 17160 based on amplicon sequencing. These identified SNPs showing genome-wide physical distribution (bp) on eight chickpea chromosomes were genotyped on 94 cultivated and wild Cicer accessions using a maximum of 34-plex high-throughput MALDI-TOF Sequenom mass array genotyping assay. The optimization and genotyping of these multiplex assays on 94 accessions validated 380 SNPs with high degree of reproducibility (100%) and genotyping success rate (98.9%). For each SNP locus, a call cluster plot was generated depicting the low and high mass homozygote and heterozygous yields . The derived genotype calls of each accessions were automatically called and compared the masses of the peaks with the pre-determined masses estimated during pre-PCR amplification and single nucleotide extension primers designing. The multiplex assay of 384 SNPs was overall successful in discriminating homozygous and heterozygous alleles for 380 SNP loci , which produced 35720 genotype calls showing polymorphism among 94 accessions. The remaining four SNPs with 376 genotyping data were identified either as monomorphic or without genotype calls. The functional annotation of experimentally validated informative 334 microsatellite markers- and 380 SNP markers-containing genes revealed their maximum correspondence with TF gene families like bZIP (basic leucine zipper), bHLH (basic helix-loop-helix) and AP2-EREBP (APETALA2 ethylene-responsive element binding proteins) , followed by growth and metabolism-related enzymes and expressed proteins .A set of 496 (including 343 genic and 153 genomic microsatellite markers) microsatellite markers distributed across eight chickpea chromosomes showing reproducible amplification were genotyped in 94 cultivated and wild cessions using thcessions using thcessions using thCicer accessions . These polymorphic markers amplified a total of 3703 alleles which varied from 2 to 13 with an average of 4.26 alleles per marker locus . The PIC and gene diversity estimated for the informative microsatellite and SNP markers ranged from 0.12 to 0.98 and 0.32 to 0.99 with a mean of 0.56 and 0.60, respectively. Even though the extent of polymorphism detected by microsatellite (96.4%) and SNP (98.9%) markers among 94 accessions was comparable with each other, the microsatellite markers, however revealed higher allele numbers , PIC and gene diversity in contrast to that estimated using SNP markers (Table S4). The heterozygosity and major allele frequency among accessions detected by both microsatellite and SNP markers varied from 0.01 to 0.32 and 0.20 to 1.0 with an average of 0.02 and 0.70, respectively . Maximum heterozygosity and major allele frequency among accessions were detected by SNP markers rather than by microsatellite markers . Inter-specific polymorphism between cultivated and wild species was higher in contrast to intra-specific polymorphism within cultivated (desi and kabuli) and wild species . Intra-specific polymorphic potential detected by markers was maximum in C. reticulatum , followed by C. judaicum and minimum in C. echinospermum . Maximum heterozygosity of markers was observed in wild C. judaicum (0.035), while it was completely absent in the cultivated species. In spite of detecting low PIC, gene diversity and allele numbers by SNP markers as compared to microsatellite markers, the trend of intra- and inter-specific polymorphism observed using combined analysis of microsatellite and SNP markers among 94 Cicer accessions remained alike with that of individual marker assays. The degree of polymorphism detected by markers among the species/accessions belonging to secondary gene pool was higher compared to primary gene pool . We classified the 94 cultivated and wild accessions under study into two groups as Fertile Crescent and Central Asia (India) based on their corresponding geographical origin. Remarkably, the markers revealed much higher polymorphic potential among the species/accessions originating from Fertile Crescent in contrast to those from Central Asia .The large-scale validation and genotyping of 880 comprising of 496 microsatellite and 384 SNP markers identified 858 (97.5%) markers (including 478 microsatellite and 380 SNP markers) as informative showing polymorphism among 94 cultivated and wild Cicer accessions revealed exact correspondence of fragment length polymorphism (bp) detected by markers among seven diverse wild and cultivated Cicer accessions with expansion and contraction of their repeats . However, substitutions of single nucleotides in the sequences flanking the microsatellite repeat-motifs across seven wild and cultivated Cicer accessions was evident.The cloning and sequencing of size variant amplicons of a selected 50 microsatellite marker sets from the Cicer species utilizing the genotyping information of genome-wide well distributed 858 markers (including 478 genic and genomic microsatellite markers and 380 TF gene-based SNP markers) was estimated. The microsatellite marker-based genetic distance among 94 accessions varied from 0.11 to 0.94, while SNP marker-based genetic distance in these accessions ranged from 0.10 to 0.42. A strong correlation (R2\u200a=\u200a0.77) between genetic distance estimated by two marker types (microsatellite and SNP markers) based on allele sharing for all pairs of accessions was evident . The use of both microsatellite and SNP markers revealed a broad range of genetic distance from 0.11 (kabuli cv. ICCV96329 - kabuli cv. BG2024) to 0.94 (C. reticulatum cv. ICC17160 - C. judaicum cv. ILWC283) with an average of 0.62. Remarkably, we observed a wider range of genetic distance (0.13 to 0.78 with a mean: 0.58) among 94 accessions using genic microsatellite and SNP markers. The mean genetic distance between cultivated and wild species varied from 0.44 (C. arietinum - C. microphyllum) to 0.70 (C. arietinum - C. judaicum), while between wild species it ranged from 0.54 (C. echinospermum - C. bijugum) to 0.68 (C. reticulatum - C. judaicum) . Maximum average genetic distance was observed among the accessions within C. reticulatum (0.52), followed by C. judaicum (0.46) and minimum within C. bijugum (0.35). The cultivated desi and kabuli accessions belonging to C. arietinum had least genetic diversity as evident from their pair-wise genetic distance (0.26) . The species/accessions included under secondary gene pool (mean genetic distance: 0.46) had higher molecular diversity compared to primary gene pool (0.37). A greater genetic diversity among the accessions/species originated from Fertile Crescent (mean genetic distance: 0.47) in contrast to those from Central Asia (0.23) was evident.The pair-wise distance matrix among 94 wild and cultivated accessions belonging to seven different Cicer accessions were depicted in a neighbour-joining unrooted phylogram . The microsatellite and SNP markers clearly differentiated all the accessions from each other and resulted in a definite grouping of six clusters (I\u2013VI) according to their species and gene pools of origination with high boot strap values (91\u201397). However, no distinct differentiation among accessions was observed by use of only TF gene-based SNP markers . Even though microsatellite markers could differentiate all 94 accessions from each other, they clustered such accessions randomly into only six different groups with much deviation from their species/gene pools of origination . Interestingly, we identified a minimum of two genomic and five genic microsatellite markers and two TF gene-derived SNP marker combinations as most informative , which could discriminate all the 94 accessions representing seven different Cicer species from each other . The cultivated species C. arietinum was highly divergent from other five wild species grouped in separate clusters with 91% occurrences, but was more closer to C. reticulatum and C. echinospermum and one Indian originated accession of C. microphyllum.The genetic relationships among 94 Cicer accessions was determined using genome-wide well distributed 858 markers employing STRUCTURE. The admixture model-based STRUCTURE simulations were conducted at varying levels of possible population numbers (K\u200a=\u200a1 to 10) with 20 replications. The average LnP(D) (log-likelihood) value increased continuously with the increase in K from 1 to 10, however its most apparent inflection was obtained at one of the best replicate of K\u200a=\u200a6 . The results of population numbers (K) were further confirmed using the second order statistics of STRUCTURE by \u0394K estimation.77 A sharp peak with a maximum value of \u0394K was obtained at K\u200a=\u200a6 , which confirms the classification of 94 accessions into six genetically distinct population groups (POP I\u2013VI) with high resolution population structure , C. echinospermum (8), C. pinnatifidum (16), C. reticulatum (16) and C. judaicum (22), respectively. The STRUCTURE-based six population group assignment of 94 accessions was further comparable to their clustering patterns, species and gene pools of origination and phylogenetic relationships ranged from 0.23 between POP II and POP III to 0.89 between POP I and POP VI (Table S6). Notably, the trends of significant genetic divergence detected by FST among six population groups was consistent with that of genetic distance . The population differentiation based on AMOVA revealed that 7.5%, 4.5% and 2.6% (P<0.01) of total molecular variation in the six population groups was attributed to genetic differentiation among populations, within populations (among accessions) and within accessions, respectively. Based on the proportion of FST estimation, divergence between population groups (59%) was higher compared to that estimated among accessions within populations (41%). Among the five wild population groups, the divergence was maximum between POP V and POP VI (FST\u200a=\u200a0.80) and minimum between POP II and POP III (0.23). Higher genetic differentiation between population groups than that within populations based on AMOVA and FST in a self-pollinated crop species like chickpea is quite expected.The population genetic structure among 94 cultivated and wild tructure Figure 4Cicer accessions used in this study clearly belonged to a structured population in which about 95% of their inferred ancestry was derived from one of the model-based population and remaining \u223c5% contained admixed ancestry . Maximum admixed ancestry (\u223c13%) was observed in POP V, followed by POP II (\u223c10%) and minimum in POP I (\u223c0.1%). The POP V had highest admixed ancestry because of their maximum allelic admixtures with POP III (\u223c6.8%) and POP I (\u223c3.7%) . Likewise, we observed maximum admixtures of POP IV (\u223c9.1%) with POP II and thus its higher admixed ancestry in POP II is expected.All the 94 cultivated and wild 2) and extent of LD decay using all possible pair-combination of genome-wide well distributed 858 markers on eight chickpea chromosomes was determined among 94 Cicer accessions belonging to six population groups. In the association panel and whole population groups, average r2 of global markers varied from 0.19 in POP VI to 0.27 in POP III (mean: 0.22) and 17% global marker-pairs exhibited significant LD (P<0.001) , indicating the existence of moderate LD level in the panel of cultivated and wild accessions. Remarkably, 43% and 14% of the linked and unlinked marker-pairs revealed significant LD (P<0.001) and average r2 of linked and unlinked markers were estimated as 0.30 and 0.20, respectively. However, the trend of LD estimates and significant LD (%) observed among six population groups using global markers remained alike with that of linked and unlinked marker-pairs. The determination of LD estimates and significant LD (%) using linked marker-pairs that are physically mapped on eight chickpea chromosomes indicated maximum estimates of average r2 on chromosome 7 (0.40) and significant LD on chromosome 4 (99.4%) . In contrast to unlinked markers, the linked markers detected higher LD estimates and proportion of significant LD in all six population groups (94 accessions) and also across eight chickpea chromosomes. Further, the LD decay of 858 marker-pairs by pooling the r2 estimates across eight chromosomes and plotting their average r2 against the physical distance of equal intervals (100 kb) up to 1200 kb was determined across six population groups. The non-linear regression curve exhibited a decreasing trend of LD decay with increase in the physical distance (kb) in all the six populations . Overall, all the population groups sustained a significant level of LD up to a physical distance of 600 kb. In population groups POP I, POP II and POP III, the LD did not decay below r2\u200a=\u200a0.1 up to 800 kb physical distance. However, the genetically more diverse population groups POP IV, POP V and POP VI showed faster LD decay (500\u2013600 kb) than that of less diverse populations POP I, POP II and POP III .The LD estimates (rn: 0.22) and 17% n: 0.22) and 17% Cicer accessions is essential for enhancing the genetic potential of cultivated chickpea. Natural genetic variation scanned in the wild Cicer, which is expected to be much wider than cultivated species Cicer accessions Medicago (http://www.medicagohapmap.org) Cicer accessions using gel-based assay, fluorescent dye-labelled automated fragment analyser and MALDI-TOF Sequenom mass array.Large-scale validation and high-throughput genotyping of sequence-based microsatellite and SNP markers and their utilization in realistic estimate of genetic diversity pattern, population genetic structure and phylogenetic relationships among wild and cultivated Cicer accessions is much higher than that estimated earlier in annual and perennial wild accessions (\u223c40 accessions) using a smaller set of genomic microsatellite markers detected by genic and genomic microsatellite markers among 94 s 30\u201340% . The genCicer accessions is higher to that obtained earlier using genomic microsatellite markers and positive correlation of both PIC and gene diversity with number of alleles/locus are in line with the findings of Upadhyaya et al. Cicer accessions confirms that the allelic variation estimated in this study across diverse wild and cultivated species was due to variation in their microsatellite repeat-length, but not because of size homoplasy (insertions/deletions in the sequences flanking the repeat-motifs) of microsatellite markers as expected during inter-specific polymorphism Cicer accessions would thus have relevance for assessing the polymorphic potential and understanding the diversity pattern, population genetic structure and phylogenetics in chickpea.The inter- and intra-specific polymorphism detected by validated 478 microsatellite and 380 SNP markers among 94 Cicer accessions using genome-wide physically mapped 478 genic and genomic microsatellite markers and 380 TF-gene-based SNP markers is higher than the molecular diversity level detected previously with the EST-derived (3 to 49% Cicer accessions is relevant. The genic microsatellite and SNP markers showing high inter-specific polymorphism (98.2%) and wider genetic diversity specifically among wild and cultivated accessions could be utilized for precise identification of true inter-specific hybrids. Besides, such markers could also have utility in monitoring the introgression breeding to access the transfer of target genic regions controlling the traits of agricultural importance including abiotic and biotic stress tolerance from wild relatives to the backgrounds of cultivated gene pool for their genetic improvement. Henceforth, a wider genetic base and functional molecular diversity detected by genic markers in the expressed sequence component of genome among cultivated and wild accessions would be of significance in selection of useful diverse parental cultivated and wild accessions in cross-breeding program for the perspective of chickpea varietal improvement. Remarkably, seven microsatellite and two SNP markers were identified as most informative that could differentiate all 94 accessions representing seven cultivated and wild Cicer species from each other. Therefore, these smaller set of microsatellite and SNP markers selected by us might assay more relevant regions of chickpea genome for establishing distinctness among accessions.A wide range of genetic diversity observed in this study among 94 cultivated and wild 3 to 49% and geno3 to 49% microsatCicer accessions using both microsatellite and SNP markers with their species and gene pools of origination than that of individual microsatellite and SNP markers suggested that the functional genetic diversity assayed in our study by combining both kinds of sequence-based markers is realistic and thus would be useful in chickpea genomics and breeding when applied to a larger set of contrasting accessions. The converse genetic attributes of microsatellite and SNP markers specifically in terms of their high/low mutation rate, multi-/bi-allelic nature and abundance in genomic distribution might have complemented the level of polymorphic potential detected by each of these two marker system in 94 accessions with each other to produce a realistic estimate of molecular diversity in the chickpea genome. It is clearly evident from high degree of genetic distance-based linear correlation (\u223c80%) between microsatellite and SNP markers as also have been observed in previous studies of many crop plants including maize and rye The present study have made use of unrooted tree to decipher the phylogenetic relationships among 94 cultivated and wild chickpea accessions. The use of unrooted tree as a reliable approach for explaining evolutionary relationships among diverse crop genotypes has extensively been reported C. arietinum, C. reticulatum and C. echinospermum species into three different population groups (determined by population genetic structure) as members of primary gene pool and accessions representing from C. bijugum, C. pinnatifidum and C. judaicum species under secondary gene pool as three diverse populations was clearly evident. These observation are fairly comparable with earlier genetic diversity, population structure and phylogenetic relationship studies on the cultivated and wild Cicer accessions using morphological traits, inter-specific hybridization, karyotyping, seed storage protein and isozyme profiling ST among three wild species accessions of primary gene pool was estimated to be 0.55, whereas it was 0.60 between primary and secondary gene pools. Thus, the evaluated FST is in line with expected evolutionary relationships among the cultivated and wild species accessions belonging to primary and secondary gene pools. In most of these studies, no distinct differentiation/groupings among the accessions belonging to Cicer species of primary gene pools and also no collinear conclusive findings on origination of accessions representing C. bijugum and C. pinnatifidum either from primary or secondary gene pools was observed. In this context, our observation on distinct differentiation of all accessions included under six different annual/perennial Cicer species into six population groups based on their species/gene pools of origination is relevant. It could be due to combined use of genome-wide well distributed genic and genomic microsatellite and SNP markers that are physically mapped on eight chickpea chromosomes and thus might provide realistic estimate of diversity pattern and population structure among wild and cultivated Cicer accessions.The inclusion of accessions belonging to Cicer species belonging to POP I (C. arietinum), POP III (C. echinospermum) and POP V (C. reticulatum) as members of primary gene pools indicate that their evolutionary closeness could be due to their domestication at the archaeological sites of South Eastern Turkey about 10,000 years ago C. reticulatum accessions under POP V with accessions of C. echinospermum (POP III) and C. arietinum (POP I), we observed low genetic variability among cultivated desi and kabuli accessions of C. arientinum. It could be due to maximum adaptation-based selection pressure during the domestication of cultivated C. arientinum from wild C. reticulatum, followed by four sequential evolutionary bottlenecks which in turn narrow down the genetic base in cultivated chickpea desi and kabuli population as compared to other wild population groups. The estimation of higher genetic diversity particularly among the wild species/accessions originated from Fertile Crescent than that of cultivated accessions from Central Asia supported these findings. A maximum admixture (\u223c9.1%) between POP II (C. bijugum) and POP IV (C. pinnatifidum) is expected as they both belong to secondary gene pools. Remarkably, population genetic structure of a perennial wild accession C. microphyllum included under POP V (C. reticulatum) contained more admixed ancestry from C. arietinum (POP I), C. reticulatum (POP V), C. echinospermum (POP III) of primary gene pools rather than C. pinnatifidum (POP IV) of secondary gene pools. The perennial C. microphyllum accession used in our study is of Indian origin and thus showed higher admixtures with Indian originated POP I (C. arietinum) and its two other evolutionarily closely related population groups (POP III and POP V) of primary gene pools. The diversity, population genetic structure, domestication and phylogenetics-related information generated in this study for 94 wild and cultivated Cicer accessions have implications for many genotyping applications including association analysis and genetic/QTL mapping targeting different qualitative and quantitative traits of agricultural importance in chickpea.The existence of about 5% admix ancestry among six population groups suggests that evolutionary divergence of all six cultivated and wild chickpea was from a common ancestor. It further concurs with the earlier reports on origination, diversification and domestication of early Neolithic chickpea along with founder crops in the Fertile Crescent (eastern Mediterranean region) about 10,000\u201312,000 years ago desi and kabuli accessions of C. arietinum is expected because of four sequential evolutionary bottlenecks occurring during their evolutionary divergence and domestication with C. reticulatum crop wild relative Cicer accessions. Overall, we inferred the population-specific LD patterns by considering the genome-wide recombination rates in 94 cultivated and wild Cicer accessions. It gave us an insight about the emerging strategies that have been utilized in HapMap projects of many crop plants Higher LD estimates and proportion of significant LD obtained in all six population groups (comprising of 94 accessions) and also across eight chickpea chromosomes using the linked markers in contrast to unlinked markers reflected the direct correlation of LD patterning with physical linkage of markers on chromosomes and marker density required to cover the genomic regions. The extensive LD estimates and extended chromosomal LD decay (\u223c500\u2013600 kb) determined in the six population groups using the microsatellite and SNP markers were much higher than that reported previously in cross-pollinated Cicer species using gel-based assay, fluorescent dye labelled automated fragment analyser and MALDI-TOF mass array. A wider genetic base (13\u201378%) and thus a higher natural allelic diversity specifically in the functionally relevant genic regions of wild chickpea was unraveled suggesting their utility as potential resource for trait enhancement of cultivated gene pool. The informative 858 mapped markers thus have potential for selecting divergent cultivated and wild accessions and identifying true inter-specific hybrids for successful introgression breeding program, which will then facilitate the transfer of target genomic/gene regions controlling important agronomic traits from wild to cultivated chickpea for their genetic improvement. Wider genetic base, distinct species/gene pool-wise differentiation, admixed domestication pattern, differential genome-wide recombination, extended LD decay (500\u2013600 kb) and extensive LD estimates (0.19\u20130.27) in a six structured wild and cultivated population were observed. The molecular diversity and LD-based information generated in our study considering the genome-wide recombination rate in six diverse wild and cultivated population gave clues to select informative markers for precise allelic diversity estimation and marker-based large-scale genotyping applications in chickpea. It will further help us to identify natural targets for chickpea genomics-assisted breeding by deriving the causal impact of natural selection on adaptation of diverse wild and cultivated chickpea populations that are domesticated across various agro-climatic regions.High genotyping success rate and intra-/and inter-specific polymorphic potential (97.5%) was detected by large-scale validation of genome-wide physically mapped 478 genic and genomic microsatellite markers and 380 TF gene-derived SNP markers in 94 annual/perennial Figure S1A representative 34-plex extension mass spectra for 94 cultivated and wild Cicer accessions obtained through MALDI-TOF mass array genotyping assay. The SNP IDs and their corresponding SNP loci are indicated on the top of each spectra.(PDF)Click here for additional data file.Figure S2Functional annotation of informative 334 microsatellite and 380 SNP markers validated in the genes showed maximum correspondence to transcription factor gene families (73.5%), followed by genes controlling growth and metabolism enzymes (23.8%) and expressed proteins (2.7%).(PDF)Click here for additional data file.Figure S3n and further corresponded with their expansion/contraction [(TTG)9/(TTG)11] of number of repeat-units.The sequencing of cloned amplicons from a microsatellite marker showing fragment length polymorphism (156 and 162 bp) among cultivated and wild accessions, validated the presence of expected repeat-motifs (TTG)(PDF)Click here for additional data file.Figure S4Correlation between genetic distances detected by microsatellite and SNP markers among 94 accessions belonging to seven wild and cultivated species. Each blue colored dot represents the genetic distance between a pair of accessions based on allele sharing of microsatellite (y-axis) and SNP markers (x-axis).(PDF)Click here for additional data file.Figure S5Unrooted phylogenetic tree depicting the genetic relationships among 94 cultivated and wild accessions belonging to seven Cicer species based on Nei's genetic distance using 380 TF gene-derived SNP markers. Molecular classification is not able to differentiate accessions into six different clusters as expected based on their species and gene pools of origination.(PDF)Click here for additional data file.Figure S6Unrooted phylogenetic tree depicting the genetic relationships among 94 cultivated and wild accessions belonging to seven Cicer species based on Nei's genetic distance using 478 microsatellite markers (including 334 genic and 144 genomic microsatellite markers). Molecular classification clearly differentiated 94 accessions into six different clusters, which corresponds to their species and gene pools of origination with deviations.(PDF)Click here for additional data file.Figure S7Optimization of number of populations varying from K\u200a=\u200a1 to 10 to determine best possible population number for 94 cultivated and wild Cicer accessions using the ad hoc procedure (A) of STRUCTURE documented by Pritchard et al. (2000) and second order statistics (delta K) (B) of Evanno et al. (2005).(PDF)Click here for additional data file.Table S1Wild and cultivated Cicer accessions used in the study and their inferred ancestry coefficients in population genetic structure analysis.(PDF)Click here for additional data file.Table S2Summary of 496 including 343 genic and 153 genomic microsatellite markers genotyped in 94 annual/perennial cultivated and wild Cicer accessions using gel-based assay and fluorescent dye labelled automated fragment analyzer.(PDF)Click here for additional data file.Table S3Summary of 384 TF gene-derived SNPs genotyped in 94 annual/perennial cultivated and wild Cicer accessions using high-throughput MALDI-TOF mass array.(PDF)Click here for additional data file.Table S4Polymorphic potential detected by 478 genic and genomic microsatellite markers and 380 TF gene-derived SNP markers among 94 cultivated and wild Cicer accessions.(PDF)Click here for additional data file.Table S5Molecular diversity detected by 478 genic and genomic microsatellite markers and 380 TF gene-derived SNP markers among 94 cultivated and wild accessions within and between Cicer species.(PDF)Click here for additional data file.Table S6ST) and genetic distance among six model-based population groups.Pair-wise estimates of genetic divergence (F(PDF)Click here for additional data file.Table S7LD estimates in six model-based individual population groups and entire population as well as across eight chromosomes.(PDF)Click here for additional data file."} +{"text": "To date, no data are available regarding native left ventricular (LV) T1 relaxation time in AF patients referred for pulmonary vein isolation (PVI). The purpose of this study was to compare native myocardial T1 value between AF patients undergoing PVI and control subjects.Native T1 mapping images were acquired using a Modified Look-Locker imaging sequence (MOLLI) in 3 short-axis planes using an ECG-triggered single-shot acquisition with a balanced SSFP readout . Late gadolinium enhanced (LGE) MRI was acquired to evaluate for myocardial scar.Thirty one AF patients referred for PVI and 17 control subjects were retrospectively identified. All patients were in sinus rhythm during the CMR scan. No AF patient had a history of myocardial infarction but 7 of 31 (23%) AF patients had history of hypertension. Non-contrast T1 was significantly elevated in AF patients . Multivariate linear regression analysis selected presence of AF as an independent and significant predictor of elevated native T1 after adjustment for age and gender .LV ejection fraction was similar between groups . No LGE myocardial scar was observed in any AF patient or control subject. Myocardial native T1 between AF patients referred for PVI. These data suggests that T1 mapping may be useful for early detection of subclinical LV myocardial abnormalities in the AF population.Our study suggests that there are subclinical differences in native myocardial TShingo Kato, MD receives scholarship from Banyu Life Science Foundation International."} +{"text": "LMNA gene mutations to evaluate clinical and molecular features associated with different phenotypes. To this purpose we included 90 LMNA-mutated myopathic patients and 36 LMNA-mutated familial cases without muscle involvement. Among the myopathic patients LGMD1B was by far the most frequent phenotype, observed in 43 (48%) patients, followed by L-CMD in 21 (23%), EDMD2 in 20 (22%) and an atypical myopathy in 6 (7%). The different myopathic phenotypes shared a similar cardiac impairment. On the other hand comparing cardiac features between myopathic and familial cases without muscle involvement we observed that cardioverter defibrillator or pacemaker were implanted more frequently in myopathic patients (n=43) (p=0.006). In addition heart transplantation and death were observed only in myopathic subgroup, respectively in 8 (9%) and 10 (11%) patients. In conclusion LMNA-related myopathies represent a continuum clinical spectrum; their clinical course appears to be dominated by cardiac involvement, considering the relatively low frequency of other complications, including loss of ambulation, assisted ventilation, surgery for scoliosis or gastrostomy. Longitudinal studied are needed to better investigate their natural history and provide indications for early management of heart involvement, in particular in first decades of life, to prevent the risk of fatal events.We conducted a retrospective study in a large cohort of myopathic patients carrying"} +{"text": "Intracellular calcium signaling is critical for initiating and sustaining diverse cellular functions including transcription, synaptic signaling, muscle contraction, apoptosis and fertilization. Trans-membrane 203 (TMEM203) was identified here in cDNA overexpression screens for proteins capable of modulating intracellular calcium levels using activation of a calcium/calcineurin regulated transcription factor as an indicator. Overexpression of TMEM203 resulted in a reduction of Endoplasmic Reticulum (ER) calcium stores and elevation in basal cytoplasmic calcium levels. TMEM203 protein was localized to the ER and found associated with a number of ER proteins which regulate ER calcium entry and efflux. Mouse Embryonic Fibroblasts (MEFs) derived from Tmem203 deficient mice had reduced ER calcium stores and altered calcium homeostasis. Tmem203 deficient mice were viable though male knockout mice were infertile and exhibited a severe block in spermiogenesis and spermiation. Expression profiling studies showed significant alternations in expression of calcium channels and pumps in testes and concurrently Tmem203 deficient spermatocytes demonstrated significantly altered calcium handling. Thus Tmem203 is an evolutionarily conserved regulator of cellular calcium homeostasis, is required for spermatogenesis and provides a causal link between intracellular calcium regulation and spermiogenesis. Transmembrane protein 203 (TMEM203)\u2013[Hs_00540709_s1]. For TG and Iono treated MEF cells the following taqman gene expression assays were used\u2014Careticulin [Mm_00482936_m1], Calcitonin receptor [Mm_00516989_m1].Same RNA preparations were used for microarray study and data confirmation. 2ug of RNA was reverse transcribed using High Capacity cDNA RT kit (Applied Biosystems). For expression analysis in Mouse tissue\u2019s cDNA were purchased from Clonetech (Mouse MTC panel). Real time PCR reactions were performed using TaqMan Universal PCR Master mix in an ABI7500 Fast Real-Time PCR system as per manufacturer's instructions (Applied Biosystems). Average threshold values (CT) from three to four PCRs were determined for target genes, and these values were normalized to average GAPDH (\u0394CT). Changes in gene expression were calculated as-fold change compared with control samples using the comparative \u0394\u0394CT method. Taqman gene expression assays : transmembrane protein 203 domains, short N- and C-terminal domains with no obvious functional domains . domains . The preCellular localization of a TMEM203-GFP fusion protein was examined by imaging with ER, mitochondria and plasma membrane (PM) markers. Based on the linescan co-lozalization tool TMEM203-GFP was predominately co-localized with the ER marker protein in HeLa cells . ConsideTMEM203 might increase cytoplasmic calcium levels via either directly activating SOCE or via lowering ER calcium stores. To distinguish between these models, we first examined ER calcium stores indirectly by measuring the increase in cytoplasmic calcium levels after release of ER stores. HEK293 cells stably containing an IPTG-inducible TMEM203 construct were treated with various levels of IPTG and the ER flux into the cytosol was measured in the absence of extracellular calcium after exposure to thapsigargin (TG) or ionomycin. IPTG induced TMEM203 expression resultedTmem203 null MEF cells as compared to Het or WT MEFs (Tmem203 null MEF cells (TMEM203 deficient mice were created See and ER c WT MEFs . TreatmeEF cells . These dCarl) and calcitonin receptor [Carl and Calcr were induced as previously reported. Induction of Carl and Calcr were significantly reduced in Tmem203 null MEF cells as compared to WT-MEF cells ,42 upon EF cells indicatiTmem203 null male mice failed to produce any litter during the duration of the experiments. Female Tmem203 null mice were fertile although the total numbers of litters (12 litters) were lower than that produced by WT or heterozygous females .Tmem203 null male and female mice were born at normal Mendelian frequency and showed no gross morphologic alterations, except for a ~10% reduction in weight. While heterozygotes were fully fertile, homozygous mating pairs produced no offspring despite the presence of copulatory plugs. Uninterrupted mating between mice of different genotypes were followed for over a period of 17 weeks to deterTmem203 null mice by light microscopy (data not shown). Further, the serum levels of Testosterone, follicle stimulating hormone and luteinizing hormone (LH) in wild type and Tmem203 null mice were comparable (data not shown).Tmem203 expression, determined by quantitative real-time PCR from various tissue RNAs, was most abundant in testes , consistTmem203 heterozygous mice (data not shown).Tmem203 deficient mice lacked mature spermatozoa (complete azoospermia) as detected by a computer assisted sperm analyzer CASA: . HistoloTmem203 null mice, spermatogenesis was examined in detail. We first evaluated the distribution of propidium iodide stained testicular cells from 35 day old or 30 week old Tmem203 null and wild type mice. DNA staining during the first wave of spermiogenesis at day 35 was indistinguishable from WT mice an overall but subtle reduction in numbers of late stage post-meiotic spermatids (steps 9\u201316) in the stages of the cycle of spermatogenesis in which progressive elongation and condensation of the nucleus occurs [Tmem203 null mice in non-spermatogenic cells including Sertoli cells and interstitial Leydig cells. In Tmem203 null mice, atrophy of individual seminiferous tubules was occasionally observed multifocally scattered randomly throughout testes (data not shown).Microscopic evaluation of the testes harvested from either 38-week-old or 48-week-old male mice showed prominent defective spermiogenesis in Tmem203 deficient mice, despite a relatively normal overall structural appearance and organization of the seminiferous tubular epithelium . The pres occurs ,31, b) dTransmission electron microscopy performed on testes harvested from 32-week-old Tmem203 null mice and age-matched wild type mice confirmed changes observed by bright field light microscopy including degenerative intra-cytoplasmic vacuolar changes as well defective spermiation. In addition, ultra-structural evaluation of the testes from Tmem203 null mice revealed abnormalities associated with the shape of the head of individual elongated spermatids (teratozoospermia) despite apparent normal condensation of chromatin and morphology of the acrosome . NumerouTmem203 wild type and null mice showed significant changes in testes and ATPase, Ca++ transporting, plasma membrane 1 (Atp2b1 / Pmca1) RNAs and reduction ofTrpv6, Trpm5 and Trpm8 calcium channel RNAs in Tmem203 deficient testes. The differential expression of these genes was further validated by real-time quantitative RT PCR . The mechanism by which TMEM203 overexpression releases ER calcium stores is not clear. Exogenously expressed TMEM203 was localized to the ER and was TMEM203 depletion, either via siRNA or through gene disruption also lowered ER calcium stores . Thus, aIn this regard it is worth noting that other proteins, notably of the BCL-2 family, have been reported to have bidirectional/opposing effects on ER calcium stores. For example, overexpression of BCL-2 lowers ER calcium stores by several reported mechanisms including acting as a leak channel, inhibiting refilling by SERCA and by enhancing activation and calcium release by the IP3R . Conversin vitro with primary spermatocytes. The altered expression of calcium pumps and channels might be due to either an attempt by cells to compensate for loss of Tmem203 or indirectly due to developmental defects. It is worth noting that loss of Bcl2 also appears to result in a dramatically increased rate of calcium extrusion in pancreatic acinar cells due to increased function of the Pmca calcium pump [Tmem203 deficient male mice were infertile with a failure to complete spermatogenesis and spermiogenesis. At present, morphologic analysis does not pinpoint a single aspect of spermatogenesis affected by TMEM203 deficiency. Haploid spermatids appear to be reduced in numbers and spermiogenesis fails after nuclear condensation, resulting in a large reduction in elongated spermatids, and accumulation of small numbers of degenerative malformed sperm heads which fail to undergo spermiation .While thium pump .C elegans lacking expression of mammalian homologs of calreticulin (Crt-1)\u2014the calcium binding chaperone protein or calcineurin (Cna or Cnb)\u2014the calcium dependent phosphatase are sterile due to defects in sperm development in addition to oocyte development and fertilization [A clear role for calcium stores in spermiogenesis has not been directly described although a number of genes regulated by calcium have been shown to be essential for sperm development. lization ,51Tmem203 null mice was described with Calcium and integrin binding 1 (CIB1)/calmyrin null mice [A similar spermiogenesis defect as in ull mice . CIB1 isull mice . It willull mice and as mull mice suggestsThe defective calcium handling in testicular cell preparations, which were predominantly haploid spermatocytes , suggestTmem203 is a highly conserved gene and is absolutely requirement for fertility. Tmem203\u2019s role in maintaining steady state ER calcium levels suggests it may modulate a variety of biological processes. Indeed, preliminary analysis suggests that TMEM203 deficiency also results in other phenotypes. TMEM203 deficient mice are about 10% smaller than WT siblings, have mild immunologic defects at least in vitro, and exhibit evidence for impaired neurologic function, though these phenotypes have not been sufficiently characterized for publication yet. The absolute requirement for TME203 in fertility would seem to be sufficient to explain its high level of evolutionary conservation. Tmem203 null mice may serve as an excellent tool to understand the regulation of intracellular calcium levels and its interdependence on carefully orchestrated modulation of ER calcium stores. Tmem203\u2019s role in spermatogenesis adds to the complexity of this fascinating process and strongly suggests a role of intracellular calcium homeostasis during mammalian spermatogenesis.S1 FigTMEM203-FLAG over-expression in 293 cells for 48 hrs induced NFAT dependent luciferase expression in a dose dependent manner, Treatment with 1uM PMA for 8 hrs enhanced TMEM203 mediated NFAT activation whereas a treatment with 5uM cyclosporine A or 10 nM SKF96365 inhibited TMEM203\u2019s activity. Data is representative of at least 3 independent experiments .(TIF)Click here for additional data file.S2 Fighttp://www.ibi.vu.nl/programs/pralinewww/). The following TMEM203 protein sequences were retrieved from NCBI\u2014Homo sapiens NP_444273; Mus musculus NP_796318; Xenopus laevis NP_001085810; Macaca mulatta NP_001248131; Gallus gallus XP_001234196; Danio rerio NP_001002519.(A)-Trans-membrane domain prediction of the TMEM203 sequence of using the TMHMM algorithm. TMEM203 is predicted to have 4 pass TM domains (red) with 3 loops and both N terminal and C terminal (green) facing the same side. (B)- TMEM203 amino acid sequences from various organisms were analyzed for conservation using PRALINE multiple sequence alignment tool. ((TIF)Click here for additional data file.S3 FigWestern analysis of complexes immune-precipitated with endogenous STIM1 from HEK293 cells with indicated antibodies shows specific interaction with overexpressed TMEM203-FLAG. (Representative of atleast 2 independent experiments).(TIF)Click here for additional data file.S4 FigTmem203 cDNA from lung tissue. The 102 bp fragment from the vector (marked in bold and underlined) disrupts the adjoining Tmem203 gene sequence in the Tmem203 null genome. The disrupted Tmem203 gene in the Tmem203 null genome lacks the start codon (ATG) for Tmem203 orf. While antibody adequate for detection of TMEM203 protein are not available, sequencing of cDNAs demonstrate that remaining tmem203 mRNA from knockout animals could only encode the last 38 amino acids of the predicted tmem 203 protein.(A)- As described in the methods section. WT: wild type allele; KO+neo: knockout allele with neo; KO: knockout allele without neo. B: BamHI. (B). Gene disruption was confirmed by sequencing (TIF)Click here for additional data file.S5 Fig(A) MEF cells derived from Tmem203\u2014WT and null mice were stimulated with 100 nM TG for 4 hrs and subsequently the expression of calcium/NFAT dependent genes\u2014Calreticulin or Cacitonin receptor (Caclr) were determined by quantitative real-time PCR. . Significant difference in Carl expression and Caclr expression was observed between WT and Tmem203 null MEF cells. (B) As described in (A), the expression of Carl and Caclr was determined in response to stimulation with 1uM ionomycin. Significant difference in Carl expression and Caclr expression was observed between WT and Tmem203 null MEF cells.(TIF)Click here for additional data file.S6 FigQuantitative real-time PCR analysis of Tmem203 transcript from various mouse tissue types showing abundant expression of Tmem203 in testes.(TIF)Click here for additional data file.S7 FigTmem203 null mice had significantly lower (A) Mean absolute body weights; 32.93 \u00b1 0.80 grams versus 27.66 \u00b1 1.98 grams, respectively; p value = 0.0006. (B) Mean absolute combined left and right testes weights; 0.2216 \u00b1 0.103 grams versus 0.1608 \u00b1 0.0142 grams, respectively; p value < 0.0001. (C) Mean testes to body weight ratios; 0.0067 \u00b1 0.004 versus 0.0058 \u00b1 0.0007 grams, respectively; p value = 0.0458. (D) Mean testes to brain weight ratios; 0.4544 \u00b1 0.0570 grams versus 0.3504 \u00b1 0.0332 grams, respectively; p value = 0.0078.Compared to wild type control mice, (TIF)Click here for additional data file.S1 TableList of cDNAs, in addition to TPRV6 and PKA that induced translocation of CRTC1 to the nucleus in a HeLa CRTC1-eGFP expressing cell line. However, only TMEM203, could efficiently translocate TORC1 without inducing gross morphologic and/or apoptotic changes.(DOCX)Click here for additional data file.S2 Table2 FC < -0.5 are shown.Only genes with Avg Log(DOCX)Click here for additional data file.S3 Table2 FC >0.5 are shown.Only genes with Avg Log(DOCX)Click here for additional data file."} +{"text": "Recent reports have shown than many identically named genetic lines used in research around the world actually contain large amounts of uncharacterized genetic variation as a result of cross contamination of stocks, unintentional crossing, residual heterozygosity within original stocks, or de novo mutation. 27 public, large scale, RNA-seq datasets from 20 independent research groups around the world were used to assess variation within the maize (Zea mays ssp. mays) inbred B73, a four decade old variety which served as the reference genotype for the original maize genome sequencing project and is widely used in genetic, genomic, and phenotypic research. Several clearly distinct clades were identified among putatively B73 samples. A number of these clades were defined by the presence of clearly defined genomic blocks containing a haplotype which did not match the published B73 reference genome. The overall proportion of the maize genotype where multiple distinct haplotypes were observed across different research groups was approximately 2.3%. In some cases the relationship among B73 samples generated by different research groups recapitulated mentor/mentee relationships within the maize genetics community. How. How2]. e genome . Resequee genome . The proe genome .Here we set out to quantify how severely these issues of divergence among samples labeled as belonging to the same genetic background impact maize (Zea mays), a preeminent model for plant genetics over the past century. Unlike soybean and arabidopsis, maize is a naturally outcrossing species, so reference genotypes must be maintained by manually controlled self-pollination in each generation. Previous studies using small sets of individually scored markers have identified genetic variation between different sources of the same maize inbred . This stA search of NCBI\u2019s sequence read archive identified 25 Illumina RNA-seq data sets deposited by 19 independent research group in three countries . Two addftp://ftp.ensemblgenomes.org/pub/plants/release-22/fasta/zeamays/dna/) using GSNAP in version 2014-12-29 [52 [5248] [In soybean, the published reference genome was found to consist of a mosaic of sequences observed in two separate sources of the reference variety and likely is not representative of the haplotype present in any individual plant . In maizde novo mutations, segregation of heterozygous loci in the original B73 founder accession [The interspersed SNPs distributed over ten chromosomes of maize may result from ccession , or fals~1% of the total maize genome) likely represents residual sequence from the knotted1 mutant donor parent line and is consistent with at least 5 generations of backcrossing (expected contribution of the donor parent = ~1.56%). Similar accidental fixations of unlinked regions may have occurred during the intentional introgression of other traits into a B73 background, such as disease resistance genes [Instead we propose a model based on the results from Sample USA 12. USA 12 consists of homozygous wild-type plants selected from family segregating for the Knotted1 . Therefoce genes .The monophyletic placement of Chinese B73 datasets suggests that the B73 seed available in China likely originated from a single transfer from the USA, apparently of seed belonging to the USA North clade and is an indicator of current tight controls on seed import/export which limit the ease with which seed change be exchanged between collaborators in China and the United States. Samples from Germany did not consistently form a monophyletic group. The concordance of academic lineages and genomic relationships in the UC Berkeley subclade acts as a notable positive control. More extensive sampling of B73 samples from many labs which employ this genotype in maize genetics research but have not, to date, published RNA-seq datasets may identify further B73 clades and subclades and additional cases where specific genomic variations have dispersed across the country as graduate students and postdocs leave a given lab for faculty positions of their own.The existence of genomic variation among samples labeled as belonging to the same accession creates barriers to reproducibility, one of the core requirements of the scientific method . In thisS1 Fig(TIF)Click here for additional data file.S2 Fig(TIFF)Click here for additional data file.S3 Fig(A) The maximum likelihood phylogenetic tree of 27 data sets by imputed SNP set; (B) One most parsimonious phylogenetic tree of 27 data sets by imputed SNP set.(TIF)Click here for additional data file.S4 Fig(TIFF)Click here for additional data file.S5 Fig(TIF)Click here for additional data file.S6 Fig(TIF)Click here for additional data file.S7 Fig(TIFF)Click here for additional data file.S8 Fig(TIF)Click here for additional data file.S9 Fig(TIFF)Click here for additional data file.S10 Fig(TIF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file."} +{"text": "Natural killer activity is believed to be important contributor of a patient\u2019s immune system to fight cancer. However, cancer patients have reportedly defective NK activity and the malignant target frequently has developed mechanisms to escape detection of NK cells. Our research is aimed at overcoming this NK cell deficiency.Malignant autologous epithelial cells of 10 colorectal carcinoma patients were separated by cell culture procedures. Peripheral blood mononuclear cells (PBMCs) were stimulated with their mitomycin treated autologous tumour cells or allogeneic SW742 colorectal carcinoma cell line. The expression of CD3, CD56, NKG2D and NKp44 were detected with flowcytometry and reverse transcription-PCR. NK activity of PBMCs against K562 target cell line was measured by MTT colorimetric assay.Stimulation with autologous tumour cells and allogeneic SW742 colorectal carcinoma cell line augmented CD56+ and CD56+CD3+ cells and up-regulated NKG2D and NKp44 expression. NK activity of PBMCs after co-incubation with autologous tumour cells or SW742 was significantly raised.Our results demonstrated that stimulation of PBMCs by SW742 can significantly improve NK activity as much as by autologous tumour cells which was confirmed by the higher expression of NKp44 and NKG2D. Since the separation of autologous tumor cells is difficult and time consuming the allogeneic tumour cell line could be a good replacement for large scale short term generation of activated NK cells. These data may help to improve cancer immunotherapy protocols."} +{"text": "Maternal lower genital tract during pregnancy is a complex niche of microbes that normally inhabit or cause infections in few instances. Association of various microbial flora and adverse pregnancy outcomes is being increasingly explored. The study was aimed to determine the prevalence of lower genital tract infections (LGTI) among pregnant women and to determine the common etiologies of LGTI and their association with adverse pregnancy outcomes, Pre term birth (PTB) and Low birth weight (LBW).A hospital based observational cohort study comprising 486 asymptomatic, antenatal women in the age group of 18-35 and the gestational period of 8-24 weeks was carried out. High vaginal and endocervical swabs were tested for Nugent\u2019s score, culture and PCR for Mycoplasmas and Chlamydia. All the women were followed until delivery.G.vaginalis (2%), Candida sp (16%), anaerobic GNB (12%), anaerobic GPB (9%) and Mycoplasmas (8%). Prevalence of PTB and LBW were; 30(6%) and 59(13%) respectively. Presence of anaerobic GNB and GPB (Nugent\u2019s Grade II organisms) was statistically significant with both PTB and LBW .Prevalence of LGTI s among study population was 134(28%) comprising BV (2%), candidiasis (13%) , trichomoniasis (8%) and anaerobic vaginitis (9%). Nugent\u2019s scoring delineated study subjects as Grade I (84%), Grade II (14%) and Grade III (2%). Prevalence of various microbes were; Incorporation of a simple diagnostic modality like Nugent\u2019s scoring system in antenatal care can help in early diagnosis and management of asymptomatic LGTIs; thereby reduce associated adverse pregnancy outcomes."} +{"text": "HIV induced endothelial dysfunction leads to premature atherosclerosis which may manifest as peripheral arterial disease (PAD) of lower limbs. Studies are not available on prevalence of PAD among antiretroviral therapy (ART) na\u00efve HIV infected population. Our objective was to explore the prevalence of PAD among HIV seropositive cases, and to determine relation of PAD with HIV infectivity and its correlation with CD4 count.This hospital based cross sectional study included 100 consecutive newly diagnosed HIV seropositive ART na\u00efve cases (age 20-49 years) and 100 age and sex matched HIV seronegative controls. PAD was diagnosed by Doppler study of lower limb arteries.Prevalence of PAD was significantly more among HIV infected cases (71%) in comparison to HIV negative controls (p <0.001). Among the cases of PAD, 53% (38/71) patients were asymptomatic. Active tuberculosis was present in 25% (18/71) of HIV infected PAD patients. The mean CD4 count among the HIV positive PAD cases was 220 cells/ dl. HIV sero-positivity was found to be independently contributing to development of PAD. Male gender, concurrent tuberculosis and low CD4 count came out to be individually contributing to PAD among HIV sero-positive cases in multivariate analysis.Prevalence of symptomatic and asymptomatic PAD was high in the ART na\u00efve HIV-infected cases of relatively younger age group. HIV infected patients should undergo evaluation for lower limb peripheral arterial diseases in appropriate background. Larger population based prospective study involving HIV seropositive patients would enlighten the temporal association of PAD and other cardiovascular morbidities."} +{"text": "Low back pain (LBP) is one of the most common musculoskeletal disorders . Exercis21 patients (age: 35.5\u00b11.4 years) with LBP were randomly assigned to a rollover footwear and lumbar extension exercise group (n=11) or a lumbar extension exercise only group (n=10). Baseline and 4 weeks post intervention measures were pain , disability (Oswestry LBP disability) and lumbar posture when standing barefoot .Participants in both groups showed significant decreases in pain levels and disability after four weeks. Participants in the shoe and exercise group had significantly greater decreases in pain (p=0.04) and demonstrated 11.8% greater reduction in disability, but this did not reach statistical significance (p>0.05) (Table This result suggests that the rollover footwear could be part of a treatment protocol for greater reduction in pain level in patients with LBP. However, the effects on lumbar biomechanics and association with changes in pain and disability remain unclear and requires further investigation."} +{"text": "Mycobacterium, yielded novel enzymes and tools for the genetic manipulation of mycobacteria. We report here the complete genome sequences of nine mycobacteriophages, including a new singleton, isolated using Mycobacterium smegmatis mc2155 as a host strain.Genome analyses of a large number of mycobacteriophages, bacterial viruses that infect members of the genus The analysis of those sequences has revealed impressive phage diversity, generating 21 clusters (A\u00b7U) of which the phages belonging to the same group have at least 50% nucleotide similarity , Argentina, by whole-genome shotgun sequencing using a Life Sciences GS-FLX 454 sequencer. When the genomes had defined ends, these were determined by PCR and Sanger sequencing of the product, as previously reported (http://cobamide2.bio.pitt.edu), while probable function assignment was done with HHpred and Pfam (5orf47) encoding a putative integrase belonging to the tyrosine recombinase family. Bioinformatics analysis suggested a possible tRNA gene (MSMEG_5758) for the integration of this mycobacteriophage into the M.\u00a0smegmatis chromosome. Roughly 50% of the orf genes of 32HC displayed homology to Mycobacterium abscessus genes encoding hypothetical proteins of unknown function; however, preliminary experiments indicated that 32HC does not infect this mycobacterial species.We isolated >40 novel mycobacteriophages from soil samples of several geographic locations in Argentina using nd Pfam 58. Three KJ028219 (32HC), KJ192196 (40AC), KJ410132 (20ES), KJ410133 (Jolie2), KJ410134 (CRB1), KJ433974 (Hosp), KJ433973 (39HC), KJ433975 (40BC), and KJ433976 (Jolie1).The complete genomes of these nine mycobacteriophages have been deposited in GenBank under the following accession no.:"} +{"text": "DNAH6 can cause heterotaxy and ciliary dysfunction similar to PCD. We provide the first evidence that trans-heterozygous interactions between DNAH6 and other PCD genes potentially can cause heterotaxy. DNAH6 was initially identified as a candidate heterotaxy/PCD gene by filtering exome-sequencing data from 25 heterotaxy patients stratified by whether they have airway motile cilia defects. dnah6 morpholino knockdown in zebrafish disrupted motile cilia in Kupffer\u2019s vesicle required for left-right patterning and caused heterotaxy with abnormal cardiac/gut looping. Similarly DNAH6 shRNA knockdown disrupted motile cilia in human and mouse respiratory epithelia. Notably a heterotaxy patient harboring heterozygous DNAH6 mutation was identified to also carry a rare heterozygous PCD-causing DNAI1 mutation, suggesting a DNAH6/DNAI1 trans-heterozygous interaction. Furthermore, sequencing of 149 additional heterotaxy patients showed 5 of 6 patients with heterozygous DNAH6 mutations also had heterozygous mutations in DNAH5 or other PCD genes. We functionally assayed for DNAH6/DNAH5 and DNAH6/DNAI1 trans-heterozygous interactions using subthreshold double-morpholino knockdown in zebrafish and showed this caused heterotaxy. Similarly, subthreshold siRNA knockdown of Dnah6 in heterozygous Dnah5 or Dnai1 mutant mouse respiratory epithelia disrupted motile cilia function. Together, these findings support an oligogenic disease model with broad relevance for further interrogating the genetic etiology of human ciliopathies.Heterotaxy, a birth defect involving left-right patterning defects, and primary ciliary dyskinesia (PCD), a sinopulmonary disease with dyskinetic/immotile cilia in the airway are seemingly disparate diseases. However, they have an overlapping genetic etiology involving mutations in cilia genes, a reflection of the common requirement for motile cilia in left-right patterning and airway clearance. While PCD is a monogenic recessive disorder, heterotaxy has a more complex, largely non-monogenic etiology. In this study, we show mutations in the novel dynein gene DNAH6 as a novel gene that can cause both heterotaxy and PCD. We further showed DNAH6 can interact with other PCD genes to mediate a more complex oligogenic etiology of disease. Thus experimental modeling with double gene knockdown showed digenic interactions of DNAH6 with DNAH5 or DNAI1 could disrupt motile cilia function in the respiratory epithelia and also cause heterotaxy in zebrafish embryos. These findings provide the first experimental evidence indicating oligogenic interactions can contribute to the complex genetics of heterotaxy.Heterotaxy is a birth defect involving randomization of left-right body axis. Its genetic etiology is still poorly understood, but recent studies suggest mutations in genes causing primary ciliary dyskinesia (PCD), a sinopulmonary disease, also can cause heterotaxy. Moreover, heterotaxy patients can show airway cilia dysfunction reminiscent of PCD. The link between these two seemingly disparate diseases reflects the common requirement for motile cilia in both left-right patterning and airway mucus clearance. Sequencing analysis of heterotaxy patients together with experimental modeling identified Heterotaxy and primary ciliary dyskinesia are both rare heritable disorders with prevalence of approximately 1 in 10,000[A link between PCD and heterotaxy is further supported by the recent finding of a high prevalence of airway ciliary dysfunction similar to that seen with PCD in heterotaxy patients with congenital heart disease (CHD). This is of clinical importance, as heterotaxy is highly associated with complex CHD and CHD/heterotaxy patients are known to have high postsurgical morbidity and mortality associated with more respiratory complications. These fDNAI1 mutation known to cause PCD. This would suggest a more complex genetic model of disease mediated by heterozygous mutations in multiple PCD/cilia related genes, ie. trans-heterozygous interactions. Such a model of disease is attractive, given ciliogenesis is a complex multi-step biological process involving hundreds of proteins and mediated by many large multiprotein complexes. As yet, such multigenic interactions in heterotaxy have not been experimentally investigated.PCD is a monogenic recessive disorder that is genetically heterogeneous with over 30 PCD genes identified. These mostly encode proteins in cilia or are required for motile cilia function. As these genes only account for approximately 60% of PCD cases. While hDNAH6 as a new heterotaxy/PCD candidate gene. We showed DNAH6 knockdown disrupted motile cilia function in the human and mouse airway, and in zebrafish Kupffer\u2019s vesicle. Furthermore, Dnah6 knockdown caused heterotaxy in zebrafish embryos. We further showed dual haploinsufficiency of Dnah6 and either Dnai1 or Dnah5 to model trans-heterozygous Dnah6/Dnai1 and Dnah6/Dnah5 interactions can cause airway ciliary dysfunction and heterotaxy. These findings show DNAH6 is required for motile cilia function mediating airway clearance and left/right patterning, and this may involve oligogenic interactions that can contribute to the complex genetics of heterotaxy and PCD.In the present study, we functionally tested the potential contribution of an oligogenic model of disease in heterotaxy, focusing our analysis on PCD/cilia-related genes. Exome sequencing analysis of heterotaxy patients identified dynein gene DNAH6 as the only ciliome candidate gene, with two heterotaxy patients with CD identified with heterozygous DNAH6 mutations and 12 exhibit normal motile cilia function, referred to as having no-CD [utations . DNAH6 id mother .Dnah6 transcripts are also highly expressed in the mouse embryonic node where motile cilia play an important role in establishing the left-right axis knockdown analysis in zebrafish embryos. We observed dnah6 MO knockdown caused a constellation of phenotypes associated with heterotaxy. This included abnormal curved body axis . Similarhomolog) . We demonscripts .dnah6 is required for motile cilia function and the specification of the left-right body axis in the zebrafish embryo. We note it was not possible to directly test for MO rescue of the various human DNAH6 mutations, given the very large size of the DNAH6 coding sequence precluded construction of a dnah6 expression construct. Nevertheless, together these ex vivo and in vivo gene knockdown analyses demonstrated DNAH6 is essential for motile cilia function and its deficiency can cause airway ciliary dysfunction and heterotaxy.Analysis of Kupffer\u2019s vesicle (KV), the zebrafish embryonic structure with motile cilia that is equivalent to the mouse embryonic node, revealed no change in the total number of cilia , but ratDNAH6 mutations in heterotaxy, we conducted sequencing analysis of an additional 149 heterotaxy patients. This included whole exome sequencing analysis of 23 patients from Cincinnati Children\u2019s Hospital had no effect on ciliogenesis or cilia motility (Dnai1 or Dnah5 mutant mice resulted in very sparse ciliation and dyskinetic ciliary motion (Dnah6 siRNA dose (50nM) was used to ablate Dnah6 function in wildtype respiratory epithelia , and is the ortholog of Chlamydomonas DHC2 encoding the heavy chain of inner arm species d[Chlamydonomas heavy chain gene is known, this species is missing in several mutant strains with defects in non-motor subunits. Our EM analysis showed DNAH6 knockdown caused unexpected central pair defects in human respiratory cilia. The mechanism for this defect remains unknown, but it is worth noting coupling of IDA and central pair defects has been observed in PCD patients with CCDC39 and CCDC40 mutations [species d precluded the pursuit of such experiments. An oligogenic model of disease is yet to be demonstrated clinically for heterotaxy or PCD, but there are several reports of PCD patients with only a single heterozygous pathogenic PCD mutation, reminiscent of the pathogenic DNAI1 mutation found in heterotaxy patient 9002 [BBS genes in Bardet-Biedl syndrome and digenic inheritance of RDS and ROM1 loci in retinitis pigmentosa[CEP290, RPGRIP1L, AHI1 and KIF7 have been reported to act as genetic modifiers in a spectrum of ciliopathies[An oligogenic model of disease for heterotaxy is compelling, given the complexity of ciliogenesis. Furthermore, this is consistent with clinical studies showing heterotaxy has largely a complex multi-genic etiology. Our analysis has been focused on assaying the effects of deficiency or haploinsufficiency, since most PCD mutations are in fact loss of function, such as the ent 9002 , 18, 26.igmentosa, 28. Addiopathies. While tiopathies. Finallyiopathies.DNAH6, as a candidate gene for heterotaxy and PCD. Our experimental modeling showed for the first time, DNAH6 is essential for motile cilia function mediating left-right patterning and in airway mucociliary clearance. Our findings suggest DNAH6 can act both in a recessive manner and also possibly in trans-heterozygous interactions with other PCD genes to mediate more complex oligogenic model of disease. Overall, these findings have broad relevance for interrogating the complex genetics of heterotaxy, and other ciliopathies. More insights into the genetic etiology of heterotaxy may have clinical translational application in allowing better patient stratification to identify those at risk for post-surgical respiratory complications related to airway ciliary dysfunction. This can help optimize clinical care and improve the prognosis of high-risk heterotaxy/CHD patients.In conclusion, functional analysis of airway ciliary motion in heterotaxy patients identified the novel dynein, Patient subjects in this study were recruited at Children\u00b9s National Medical Center, Cincinnati Children\u00b9s Hospital, Tokyo Women's Medical University, and Children\u00b9s Hospital of Philadelphia. All of the human subject recruitments were conducted with human study protocols approved by the respective Institutional Review Board (IRB) for the Protection of Human Subjects. Informed consents and assents were obtained at each of the recruitment sites as per approved IRB protocols at the respective institutions. All animal experiments were conducted with approved IACUC protocols at University of Pittsburgh.25 heterotaxy patients were previously recruited from the Children\u2019s National Medical Center[Dnai1 knockout mice (MGI: 4415703) and Dnah5 mutant mice (MGI: 3722326) were housed in the mouse facility of the University of Pittsburgh Rango\u2019s Research Center. All animal experiments were conducted with approved IACUC protocols at University of Pittsburgh.Zebrafish were housed and in the Zebrafish Aquaria facility of University of Pittsburgh. DNAH6 mutation from Tokyo Women\u2019s Medical University and Children\u2019s Hospital of Philadelphia (https://www.broadinstitute.org/gatk/).Targeted exome sequencing analysis of 25 patients from Children\u2019s National Medical Center was carried out with a custom Agilent SureSelect sequence capture kit . We compadelphia , pink wewww.openbioinformatics.org/annovar) and custom scripts.Sequence variants were annotated using annovar (www.1000genomes.org) with matched race/ethnicity and NHLBI exome (evs.gs.washington.edu/) databases. Pathogenicity of missense variants were assessed by PolyPhen-2, SIFT and CADD Score algorithms[Ciliome genes were examined for novel or rare variants, defined as those with <0.8% allele frequency in the 1000 Genomes (lgorithms\u201341.DNAH6 (primer sequences will be made available upon request). PCR products from each sample were then pooled and sequenced using IonTorrent Personal Genome Machine. Sequence data was analyzed using the CLCBio Genomics Workbench software as described above. The same bioinformatics pipeline as described above for the exome sequencing analysis was used to analyze for DNAH6 sequence variants. Candidate DNAH6 coding variants identified were confirmed using Sanger capillary sequencing.PCR amplifications were carried out for all 77 exons and flanking splice junctions for Dnah6 cDNA clone obtained from OpenBioSystem (www.openbiosystem.com) was used to generate the in situ hybridization probe. In situ hybridization analyses of E8.0 mouse embryos were performed using a digoxigenin-labeled Dnah6 antisense riboprobe following published protocol[Partial mouse protocol. After sDNAH6 short hairpin RNA (shDNAH6) or non-specific scramble control shRNA (shSCRAMBLE) and incubated for 3 hrs at 37\u00b0C, after which the cells were removed with collagenase and cells placed in suspension at 37\u00b0C on an orbital shaker. By expressing the short hairpin RNA during the process of reciliation, it is possible to assay protein function required for ciliogenesis. Reciliation is usually observed in 7\u201314 days, and videomicroscopy was carried out at 200 fps using a Phantom v4.2 camera to assess ciliary motion.Nasal tissue from healthy human subjects was collected using Rhino-Probe curettage of the inferior nasal turbinate and the collected tissue was resuspended in L-15 medium for videomicroscopy using a Leica inverted microscope with a 100x oil objective and differential phase contrast optics. The collected human nasal epithelia tissue was cultured using Ultroser G medium on collagen coated plates and grown to confluence. ConflueDNAH6 knockdown, total RNA from shDNAH6 and control shSCRAMBLE treated reciliated human airway epithelial cells were isolated using RNeasy micro kit with on-column DNase I digestion method (QIAGEN). cDNA were prepared and amplified using Ovation RNAseq kit version 2 (NuGen). Real-time PCR was done using Applied Biosystem HT7900 with Power SYBR Green PCR Master Mix (Applied Biosystem) following standard protocol. The ubiquitously expressed \u03b2-actin gene and cilia-related genes DNAI1 and DNAAF2 were used as internal controls. Similar analysis was also conducted using DNAH6 siRNA treated mouse epithelial cells at different dose , with cilia gene IFT80 was used as an internal control. Similar experiments were performed to assess the expression of CCNO, MCIDAS and FOXJ1 transcripts in DNAH6 knockdown and control siRNA treated samples.To assess the efficacy of Reciliated respiratory epithelial cells were suspended in cell culture medium and were blast by homogenizer for 3s to break the spheroids. Samples were spread onto glass slides, air-dried and stored at \u221280\u00b0C until use. Cells were treated with 4% paraformaldehyde, 0.1% Triton-X 100 and 1% BSA before incubation with primary (at least 1 h at room temperature (18\u201320\u00b0C) or overnight at 4\u00b0C) and secondary (30 min at room temperature) antibodies. Polyclonal anti-DNALI1, anti-DNAH5 and monoclonal mouse anti-acetylated-\u03b1-tubulin and monoclonal mouse gamma\u2013tubulin antibodies were purchased from Sigma. Highly cross-absorbed secondary antibodies were from Molecular Probes (Invitrogen). DNA was stained with DAPI (Sigma). Immunofluorescence images were taken with a Zeiss Apotome Axiovert 200 and processed with Openlab v5.1.Reciliated human respiratory epithelia were fixed in 2.5% (v/v) glutaraldehyde in 0.1 M sodium cacodylate buffer at 4\u00b0C, washed overnight and postfixed in 1% (v/v) osmium tetroxide. After dehydration, the samples were embedded in epoxy resin. Ultrathin sections were obtained from the embedded specimen and the sections were contrasted with Reynold's lead citrate and 4% uranyl acetate in ethanol. Transmission electronmicroscopy was performed with a Philips CM10 and images were captured using a Gatan, Erlangsher CCD camera system.dnah6 and dnai1 antisense morpholinos (MO) were designed and synthesized by GeneTools, LLC .dnah6 AUG-MO 5\u2019-AAGAATACATGACCAACTTCCCTGC-3\u2019dnah6 Spl-MO E24I24 5\u2019- TGAGCGTCATCAGGCCGGACCTGTT-3\u2019dnai1-134914 5\u2019-GGCTGTTTCTCCTCAGACATTTTTC-3\u2019dnai1-80431 5\u2019- TCTTTAAATAACAAGACTGCTCCGC-3\u2019dnai1 I8E9 5\u2019- GCTTGAAATCTGAGGTTTGGGTTAA-3\u2019in situ hybridization or direct phenotype visualization under a Leica stereomicroscope and photographed using a Retiga camera (Q-imaging).MO doses ranging from 1-10ng were injected into the 2-cell stage embryos as previously described along with the standard control scrambled MO. Embryos were incubated to the desired stage, then processed for fixation and spaw, myl7 and foxa3. Whole mount immunofluorescence was performed to detect monocilia in Kupffer\u2019s vesicle as described[Zebrafish embryos were fixed with 4% paraformaldehyde and analyzed by whole mount in situ hybridization using the protocol described previously. The foldescribed. AcetylaFor imaging cilia generated flow in Kupffer's vesicle (KV), zebrafish embryos (6\u20138 somites) were removed from their chorion and positioned KV facing upwards in a shallow trough cut into a 0.020\u201d thick silicone sheet glued to the bottom of a 35mm glass bottomed culture dish and placed under a 63x immersion lens of a upright microscope (Leicia DMLFSA). Microinjection pipettes were fabricated using borosilicate capillary glass (1mm OD/0.75 mm ID) on a P-97 Flaming/Brown micropipette puller , and then beveled using a BV-10 microelectrode beveler . Micropipettes were filled with 0.20 \u03bcm fluorescent polystyrene latex microspheres and mounted onto a Leicia micromanipulator. A pneumatic PicoPump was then used to inject a small amount of fluorescent microspheres into each KV. To track microsphere movement epifluorescence microscopy was conducted using a high-speed CCD camera . Video movies of cilia generated flow were captured at a frame rate of 15 frames/sec. The ImageJ software package with MTrackJ plugin was used to manually track fluorescent bead movement within each KV. High-spDnai1 knockout and Dnah5 mutant mice were collected, diced into very fine pieces and cultured using Ultroser G medium on collagen coated 6-well plates and grown to confluence[Dnah6 siRNA ranging from 0-50nM using Qiagen RNAiMax kit to determine the subthreshold dose. The Dnah6 siRNA was obtained from Qiagen (SI04413444). Cells were incubated for 4hrs at 37\u00b0C, after which the cells were removed with collagenase and placed in suspension at 37\u00b0C on an orbital shaker. Reciliation occurs after 7 days in suspension, videomicroscopy was carried out at 200 fps using a Phantom v4.2 camera to assess ciliary motion.Tracheal tissue from both wildtype and heterozygous onfluence. ConflueS1 FigDNAH6 c.6182G>A allele, but is wildtype for the DNAI1 allele. In contrast, the mother is heterozygous for the DNAI1 IVS1+2_3insT allele, but is wildtype for the DNAH6 allele.Sanger sequencing confirmed the father of patient 9002 is heterozygous for the (TIF)Click here for additional data file.S2 FigDNAH6 transcripts in human respiratory epithelia after shDNAH6 knockdown as compared to control. This is confirmed by normalizing using expression of housekeeping gene beta-actin , as well as cilia-related genes DNAI1 and DNAAF2 .Quantitative analysis by real time PCR showed marked reduction in (TIF)Click here for additional data file.S3 FigDNAH6 knockdown show no apparent change in the distribution of DNAH5 (an outer dynein arm marker), DNALI1 (an inner dynein arm marker) or RSPH4A in the ciliary axoneme. Acetylated tubulin antibody staining was used to visualize the ciliary axoneme. DAPI staining (blue) was used to visualize the nucleus. Note image for DNAH5 staining with DNAH6 knockdown contains two ciliated cells while all the other panels contain only a single multiciliated cell.Human respiratory epithelia after (TIF)Click here for additional data file.S4 Fig qPCR analysis showed CCNO, FOXJ1 and MCIDAS transcripts were not affected in human and mouse respiratory epithelia after Dnah6 knockdown as compared to control. (C) In situ hybridization showed foxj1 expression level and pattern did not change in zebrafish embryos after dnah6 antisense morpholino gene knockdown.(TIF)Click here for additional data file.S5 Figdnah6 AUG (AUG-MO) and splicing (Spl-MO) morpholinos caused heart looping defects including right sided (R), or straight (St) heart looping phenotypes.Both (TIF)Click here for additional data file.S6 FigIn situ hybridization analysis showed dnah6 transcripts are ubiquitously expressed as maternally derived transcript in the early zebrafish embryo.(TIF)Click here for additional data file.S7 Fig(A) RT-PCR analysis confirmed dnah6 splice morpholino (dnah6 splMO) disrupted proper splicing of intron 24 in MO-injected embryos vs. control (ContMO). (B) The dnah6 AUGMO provided higher percentage of curly tail phenotype as compared to the dnah6 splice MO.(TIF)Click here for additional data file.S8 Figdnah6 AUG MO or splice MO knockdown affects the KV cilia number as compared to the control.Neither (TIF)Click here for additional data file.S9 FigDnah6 siRNA reduced Dnah6 expression to ~65% compared to the control, 50nM siRNA knockdown further reduced Dnah6 expression to less than 50%.Quantitative analysis by real time PCR showed that subthreshold knockdown with 30nM (TIF)Click here for additional data file.S1 TableTable provides HGNC gene symbols and Refseq IDs for genes used for the targeted exome sequencing analysis described in (XLSX)Click here for additional data file.S2 TableDNAH6 mutations, the detailed mutation annotation was also provided in the table.This table provides detailed phenotypic data for heterotaxy patients identified with (PDF)Click here for additional data file.S3 TableDnah6 gene knockdown.Summary of the phenotypes observed in zebrafish embryos and in mouse and human respiratory epithelia upon (PDF)Click here for additional data file.S1 MovieReciliated human respiratory epithelia treated with control shRNA knockdown displayed cilia with normal coordinated ciliary motion and normal cilia waveform. Original movie was captured at 200 fps, movie playback is 30 fps . Scale Bar = 10 \u03bcm(MOV)Click here for additional data file.S2 MovieDHAH6 shRNA displayed cilia with a range of abnormal motility defects, including cilia that exhibited abnormal dyskinetic motion to completely immotile cilia. Original movie was captured at 200 fps, movie playback is 30 fps . Scale Bar = 10 \u03bcmReciliated human respiratory epithelia treated with (MOV)Click here for additional data file.S3 Moviednah6 MO injected embryos KV cilia displayed a spectrum of motility phenotypes, ranging from more normal motion (top right) to random rotational movement (middle panels) and backward/forward motion (bottom left), to being nearly immotile (bottom right). Original movies were captured at 200 fps, movie playback is 30 fps . Scale Bar = 2.5 \u03bcmKupffer\u2019s vesicle (KV) cilia in control MO injected zebrafish embryos displayed normal rotational movement (top left), but in the (MOV)Click here for additional data file.S4 Moviednah6 MO injections, KV flow (bottom panels) was not observed. Videomicroscopy was conducted using DIC imaging (top left), and epifluorescence microscopy (top right). Scale Bar = 10 \u03bcmMicroinjection of fluorescent beads into control MO injected zebrafish embryos revealed robust rotational flow, but after (MOV)Click here for additional data file.S5 MovieDnai1 knockout mouse tracheal epithelia was much more sensitive to the effects of Dnah6 siRNA knockdown. Knockdown at 30nM Dnah6, which had no effect on wildtype tracheal epithelia, caused reduced cilia density and a spectrum of ciliary motion defects ranging from immotile to restricted dyskinetic ciliary motion in heterozygous Dnai1 knockout mouse tracheal epithelia. These are similar to the phenotypes observed with 50nM Dnah6 siRNA knockdown in the wildtype tracheal epithelia. This contrasts with the completely immotile cilia observed in the tracheal epithelia of homozygous Dnai1 knockout mice. Movie was originally captured at 200 fps, and movie playback is set at 30 fps for ease of visualization . Scale Bar = 10 \u03bcmIn wildtype tracheal epithelial cells, 30nM siRNA knockdown had no effect on ciliogenesis or cilia motility, as the respiratory epithelia continued to show high ciliation density and normal ciliary motion. However, with 50nM knockdown, cilia density was significantly reduced with many areas entirely devoid of cilia, and ciliary motion was abnormal and dyskinetic. In comparison, heterozygous (MOV)Click here for additional data file.S6 MovieDnah6 siRNA knockdown had no effect on ciliogenesis or cilia motility in wildtype (+/+) tracheal epithelial cells. In comparison, 30nM siRNA in heterozygous Dnah5 (+/m) mutant tracheal epithelia cells showed significantly reduced cilia density, and ciliary motion was largely dyskinetic/immotile. Movie was originally captured at 200 fps, and movie playback is set at 30 fps for ease of visualization . Scale Bar = 10 \u03bcm30nM (MOV)Click here for additional data file."} +{"text": "Chronic Obstructive Pulmonary Disease (COPD) in the future will be increased according to increasing air pollution by smoke from factories or vehicles and smoking habits. Asthma, chronic bronchitis, bronchiectasis, emphysema and others chronic and critical conditions require minimal invasive surgery.Video Assisted Thoracoscopic Surgery (VATS) is a minimalist operation using a fiber optic cable that is connected to a video screen television for imaging objects and surgery will be done through several small incisions like keyhole of the chest cavity. VATS advantages compared to conventional surgery is the recovery time is shorter and very useful in situations whereas conventional thoracotomy surgery is not possible due to severe conditions. The drawback with this minimal incision exposes less due to obstructed ribs, but can be overcome with proper planning beforehand through a 3D reconstruction of CT Scans Thorax.Case control studyNine cases of COPD with pneumothorax complications between 44-84 years old performed VATS Lung Volume Reduction Surgery (LVRS). Comorbidities before surgery in 4 cases, such as Diabetes Mellitus, Coronary Arteriosclerosis Disease, emphysema subcutis, ulcer pepticum. Length of Stay (LOS) 5-40 days, morbidity infection in 3 cases, 1 case of gastrointestinal bleeding, prolonged pneumothorax 2 cases, 1 recurrent pneumothorax and no mortality. Functional testing of lung vital capacity about 21.5% with mean forced expiratory volume of 20%. Spirometry evaluations during the year showed an increase of 10-20%, but the functional of daily activities showed improvement and free from oxygen dependency. Four patients developed congestion right heart began three months after the respiratory rehabilitation with one person showed moderate pulmonary hypertension. 4-year follow-up to the dismissal of contamination to cigarette smoke showed significant changes in lung function of ventilation and perfusion test in Nuclear MedicineVATS LVRS showed good outcome for selected COPD."} +{"text": "Depression is a common comorbid condition of chronic pain. The current mainstay of managing comorbid chronic pain uses combination drug therapy including opioid analgesics and antidepressants. However, the cellular mechanism underlying the comorbid interaction between chronic pain and depression remains unclear.Using rat models of genetically predisposed [Wistar-Kyoto (WKY) rats] or induced depressive behavior combined with persistent nociception, we examined whether 1) brain IDO1 (a rate-limiting enzyme in tryptophan metabolism) expression and activity would be increased in rats with genetically predisposed or induced depressive behavior, 2) rats with depressive behavior would exhibit exacerbated nociceptive behavior, and 3) brain IDO1 upregulation would be contributory to both depressive and nociceptive behaviors. Depressive behaviors were assessed by using forced swimming test, sucrose preference test, tail suspension test, and open field test. Nociceptive behaviors were assessed using von Frey filaments and hind-paw withdrawal to radiant heat stimulation. RT-PCR, Western blotting, HPLC, ELISA, immunohistochemistry and cell culture were used in the study.We demonstrate that brain IDO1 expression critically contributes to the comorbid interaction between pain and depression.(1) IDO1 expression was elevated in the hippocampus of rats with either genetically predisposed or anhedonia-induced depressive behavior. (2) Rats with elevated IDO1 expression exhibited a lower baseline mechanical and thermal nociceptive threshold, whereas depressive behavior was exacerbated in rats with persistent nociception.(3) IDO1 upregulation was mediated by the IL-6/JAK/STAT signaling, resulting in a shift of tryptophan metabolism toward the IDO pathway as well as a low serotonin content in the hippocampus. (4) Inhibition of IDO1 activity or IL-6/JAK/STAT-mediated IDO1 upregulation concurrently attenuated nociceptive and depressive behaviors in the same rats.The results indicate that brain IDO1 expression critically contributes to the comorbid interaction between pain and depression and suggest that targeting brain IDO1 activity may offer a new therapeutic method for the treatment of comorbid chronic pain."} +{"text": "With increased usage of antiretroviral drugs (ARVs) in HIV uninfected persons proper reporting on suspected unexpected serious adverse reactions (SUSARs) and continued insight into serious adverse events (SAEs) is needed for adequate information on ARVs safety in such populations.We have evaluated medical documentation of persons receiving ARVs after non-occupationally HIV exposure (nPEP) during five concomitant years (2009\u20132013). SAEs and SUSARs were evaluated by two HIV physicians and defined according to international standards. In statistical methods, Kaplan Meier survival analysis was used to estimate the probability of SAE and Cox proportional hazard models to identify independent predictors of developing SAE. Only the first SAE was included in these analyses.In total, 375 persons received nPEP. The most common reason was needle stick (43%), followed by unprotected sexual intercourse (17%), rape (10%) and first aid (10%). In 84 (22%) cases, the source patient was either known to be HIV positive or within a high risk group (active injecting drug user). In total, 170 SAEs were reported, 139 persons had only one SAE and majority developed it within first two weeks. The most frequent first SAEs were gastrointestinal disorders (22%), followed by general symptoms (9%), hypersensitivity reactions (1.6%) and CNS symptoms (1.3%). The remaining events were laboratory abnormalities of liver and kidney function, haematological disorders, other and unknown, each contributing to less than 1% of all patients. 8 (2.1%) patients have developed a SUSAR . 22 (5.9%) persons discontinued nPEP due to adverse event and 19 (5.1%) required a paid sick leave from work. In multivariate analyzes, only age was independent predictor of developing SAE .In our observation, SAEs in reaction to nPEP were frequent yet usually mild events, mostly occurring in first two weeks and rarely causing discontinuation. The only significant factor increasing the risk of SAE was age. SUSARs were rare and moderately significant. More insight into this important area is required in order to ascertain proper pharmacovigilance of ARVs usage in HIV uninfected persons."} +{"text": "Severe asthma are linked with high morbidity, significant mortality and high treatment costs. Omalizumab has been shown to decrease the risk of hospitalization or Emergency Department (ED) visits in patients with uncontrolled severe allergic asthma. We aim to describe the conditions under Omalizumab was prescribed in patients followed in a Reference Center for Severe Asthma Treatment in Nova Igua\u00e7u, Rio de Janeiro; and assess the effects of Omalizumab through the Asthma Control Test (ACT) in those patients who had at least a 16 week course.Asthmatic patients treated with omalizumab between February 2013 and June 2014 were evaluated retrospectively. The conditions under Omalizumab was prescribed and ACT improvements were evaluated.A total of 19 patients were prescribed omalizumab. Prescribing criteria were: one or more ED visits in the last year (100%); high dose inhaled corticosteroid and long-acting beta2agonist use (94.7%); systemic corticosteroid use more than 3 times the last year (89.5%); FEV<80% (78.9%); daily short-acting beta2agonist use (68.4%); fast pulmonary function deterioration after systemic corticosteroid withdrawal (52.6%); death threatening asthma exacerbation episode (42%). Seven of these patients had a 16 week course of omalizumab with a significant improvement in ACT total score in six of them (86%).In our casuistic, the main criteria omalizumab was prescribed for severe asthma was ED visits. Omalizumab promoted a significant improvement in most patients' ACT total score."} +{"text": "Midwall septal fibrosis on late gadolinium enhancement (LGE) cardiovascular magnetic resonance (CMR) imaging has been shown to be an independent predictor of adverse events in patients with non-ischemic dilated cardiomyopathy (NICM). Recent studies in other cardiomyopathy cohorts, such as Hypertrophic Cardiomyopathy, suggest that LGE extent provides incremental prognostic value over its binary presence or absence. The objective of this study was to investigate the prognostic value of septal LGE quantification for the prediction of arrhythmic events among patients with NICM.42, CircleCVI). Left ventricular volumes and function were determined from SSFP cine images. The binary presence or absence of mid-wall septal LGE was determined by an expert reader. A separate blinded observer quantified total Septal LGE mass using a \u22655SD signal threshold. Strict anatomic criteria were used to define the septal region for all analyses.A total of 122 consecutive patients with NICM , defined as a LVEF<55% and no history of myocardial infarction, revascularization, or ischemic LGE by MRI were followed for the occurrence of sudden cardiac death (SCD) or appropriate implantable cardiac defibrillator (ICD) therapy. CMR imaging was performed using a 3T MRI scanner using a standard imaging protocol. All image analysis was blindly performed using commercially available software . Septal LGE mass was significantly higher among those having an event versus those without . Multivariate analysis inclusive of the 2 variables of Septal LGE mass and LVEF showed Septal LGE mass to be an independent predictor of the primary outcome . ROC analysis of Septal LGE mass was performed (AUC 0.74) with a threshold of 2.67g providing optimal sensitivity (0.69) and specificity (0.83). Using this threshold those with high Septal LGE mass (N=29) were at a markedly increased risk of sudden cardiac death or appropriate ICD therapy (HR=16.01).Signal threshold based quantification of Septal LGE in NICM patients provides superior prediction of sudden cardiac death or appropriate ICD therapy versus its binary scoring (HR 16.0 versus 4.7). LGE quantification may be a valuable tool for the risk stratification of patients with NICM.This study was funded in part by: Heart and Stroke Foundation of Canada (NA 6488 Ontario research Fund - Imaging in Cardiovascular Therapeutics), Canada Foundation for Innovation, and Calgary Health Trust."} +{"text": "Universal screening and brief intervention (SBI) for drug use among primary care (PC) patients lacks efficacy but the efficacy of SBI for low risk drug use is unknown. This 3-arm pilot study tested the efficacy of two brief interventions (BIs) for drug use compared to no BI in PC patients with low risk drug use identified by screening.We randomly assigned participants identified by screening with Alcohol Smoking and Substance Involvement Screening Test (ASSIST) drug specific scores of 2 or 3 (consistent with low risk drug use) to: no BI, a brief negotiated interview (BNI), or an adaptation of motivational interviewing (MOTIV). BNI was a 10-15 minute structured interview conducted by health educators. MOTIV was \u00bf45 minutes with an optional booster conducted by trained master's-level counselors. Primary outcome was number of days use of self-identified main drug in the past 30 as determined by validated calendar method at 6 months. Analyses were performed using negative binomial regression adjusted for baseline use and main drug.Of 142 eligible adults, 61(43%) consented and were randomized. Participant characteristics were: mean age 41; 54% male; 77% black. Main drug was marijuana 70%, prescription opioid 10%, cocaine 15%; 7% reported injection drug use and mean days use of main drug (of 30) was 3.4. At 6 months, 93% completed follow-up and adjusted mean days use of main drug were 6.4 (no BI) vs 2.1 (BNI) and 2.3 (MOTIV) .BI (both BNI and MOTIV) appears to have efficacy for preventing an increase in drug use in primary care patients with low risk use identified by screening. These findings raise the potential that less severe patterns of drug use in PC may be uniquely amenable to brief intervention and warrant replication in a larger trial."} +{"text": "S1 TableBold type indicates parameters that are biologically significant.(DOCX)Click here for additional data file.S2 TableEl Nino Southern Oscillation (ENSO) estimates are calculated by using the average Southern Oscillation Index (SOI) values from July to April of each year. Nesting success (NS) and daily nest survival rates (DNSR) were estimated using the logistic-exposure method.(DOCX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file."} +{"text": "This study aimed to investigate the virological status in liver (both tumor and adjacent non-tumor tissue), the clinical features and the contribution of occult HBV infection (OBI) to postoperative prognosis in HBeAg-negative(\u2212) hepatocellular carcinoma (HCC) patients in China. Using quantitative TaqMan fluorescent real-time PCR assays, HBV covalently closed circular DNA (cccDNA) and total DNA (tDNA) were both quantified in 11 (HBsAg(\u2212)) and 57 (HBsAg-positive(+)) pairs of tumor tissue (TT) and adjacent non-tumor tissue (ANTT) obtained from HBeAg(\u2212) HCC patients who received no antiviral treatment and were negative for anti-HCV before surgical treatment. Of 11 HBsAg(\u2212) patients, 36% were with HBsAb(+) HBeAb(+) HBcAb(+). However, only 9% of the HBsAg(\u2212) patients were HBsAb(\u2212) HBeAb(+) HBcAb(+), which accounted for the majority (93%) in the HBsAg(+) group. TT and ANTT HBV tDNAs in 11 HCC patients with HBsAg (\u2212) and HBeAg (\u2212) were all detectable. HBV cccDNA and tDNA were all lower in the HBsAg(\u2212) group than those in the HBsAg(+) group. By Kaplan-Meier analysis, patients with OBI were associated with a lower risk of cirrhosis and better overall survival (OS). The intracellular HBV DNAs, such as HBV cccDNA and tDNA are valuable biological markers for the diagnosis of occult HBV infection in HCC patients. This would assist the clinical implementation of a more personalized therapy for viral re-activation control and improve the survival rate of OBI patients. Hepatocellular carcinoma (HCC) is the third most common cause of cancer death worldwide OBI can occur not only in individuals with anti-HBs and/or anti-HBc antibodies but also in those who are negative for HBV markers HBV covalently closed circular DNA (cccDNA) is an important intermediate in the life cycle of HBV, from which the HBV pregenomic RNA and all HBV messenger RNA transcripts originate We therefore conducted a prospective study. The primary aim was to investigate the virologic status in the liver (both TT and ANTT) among these HCC patients with HBsAg (\u2212) and HBeAg (\u2212). The second aim was to determine the clinical features and the contribution of occult HBV infection (OBI) to postoperative prognosis for HCC patients with HBsAg (\u2212) and HBeAg (\u2212) in China.This study included a HBsAg-negative group (n\u200a=\u200a11) and a HBsAg-positive (+) group (n\u200a=\u200a57) of HCC patients with HBeAg (\u2212) (between March 2007 and May 2009) who received no antiviral treatment and were negative for anti-HCV before surgical resection at the Shanghai Eastern Hepatobiliary Surgery Hospital (EHBH) in Shanghai, China. The study was approved by the Chinese Ethics Committee of Human Resources at the Second Military Medical University. All study participants provided written informed consent.The inclusion criteria were patients with no evidence of hepatitis C virus (HCV) or hepatitis D virus (HDV) co-infection; no previous antiviral treatment; complete resection of tumor with sufficient safety margin (R0) and histologically proven HCC.The HBsAg-negative group: HBsAg-negative and HBeAg-negative for at least 6 months, undetectable serum HBV DNA.The HBsAg-positive group: HBsAg-positive and HBeAg-negative for at least 6 months.The exclusion criteria included a history of liver transplantation and other malignancies, tumors of uncertain origin, metastatic liver cancer, autoimmune liver diseases, drug-related liver diseases, alcoholic hepatitis and other causes of chronic liver diseases diagnosed before enrollment.Details of patient clinical diagnosis, follow up are included in 2, 0.5 \u00b5M of forward and reverse primers, and a 0.4 \u00b5M probe. Forward and reverse primers were F1 and R1 for cccDNA amplification, respectively and F2 and R2 for total intrahepatic HBV DNA amplification, respectively. TaqMan probes were TaqP1 for cccDNA quantification and TaqP2 . HBV cccDNA and tDNA were detected using real-time polymerase chain reaction (PCR) with TaqMan fluorescent probes according to the method described by Bettina et al. with a slight modification nd TaqP2 for totaP value of <0.05 was considered as statistically significant. Details are included in All statistical analyses were two sided and performed using SPSS 17.0 for Windows . A The immunological characteristics of HBeAg(\u2212) HCC patients are summarized in In the HBsAg(\u2212) group, there were 6 HBsAb(+) patients (54%) and 5 HBsAb(\u2212) patients (46%), 5 HBeAb(+) patients (46%) and 6 HBeAb(\u2212) patients (54%), and 9 HBcAb(+) patients (82%) and 2 HBcAb(\u2212) patients (18%). However, in the HBsAg(+) group, there were only 1 patients (2%) with HBsAb(+) and 56 patients (98%) with HBsAb(\u2212),54 patients (95%) with HBeAb(+) and 3 patients (5%) with HBeAb(\u2212), and 57 patients (100%) with HBcAb(+).The paired TT/ANTT of HCC patients with HBeAg (\u2212) were stratified into different group (A\u2013E) according to the immunological characteristics, and examined for intracellular HBV cccDNA and tDNA levels . Interes10 copies/106 cells, P\u200a=\u200a0.013; ANTT: 3.35\u00b10.94 vs. 4.83\u00b11.16 log10 copies/106 cells, P\u200a=\u200a0.004). Similarly, the HBV tDNA was also significantly lower in the HBsAg(\u2212) group than in the HBsAg(+) group . However, no statistical significant difference in TT/ANTT ratios was observed between the HBsAg(\u2212) and the HBsAg(+) group .The difference in TT/ANTT HBV cccDNA and tDNA between the HBsAg(\u2212) group and the HBsAg(+) one were shown in Eleven Patients with occult HBV infection had higher albumin (ALB), well-differentiated tumors (E-S grades I and II) and a lower risk to develop cirrhosis . HoweverP\u200a=\u200a0.173 and P\u200a=\u200a0.386, By Kaplan-Meier analysis, although patients with OBI did not differ significantly in overall survival (OS) and disease-free survival (DFS) and HBeAg(\u2212) were all detectable in TT or ANTT at the time of surgical resection. There were approximately half of these patients with HBsAb(+) (54%) and with HBeAb(+) (46%), and 82% of patients with HBcAb(+) in the HBsAg(\u2212) and HBeAg(\u2212) group. We confirmed that HBV could persist in liver after the disappearance of HBsAg in individuals with previous exposure to the virus, retaining the serological footprint of HBcAb positivity with such a virologic status To clarify the virological characteristics of HBV, we detected the cccDNA levels in cancerous tissues and non-cancerous tissues. cccDNA does not take part in replication directly, because it is maintained as a stable pool inside the hepatocyte nuclei in vitro study showed that occult viral isolates could fully restore replication, transcription, and protein synthesis abilities once the viruses are taken out of the host liver microenvironment Although the cause of OBI reactivation is yet to be understood, it is necessary to consider the following factors: the host's immune surveillance, restored virus, the impact of liver cancer cells and coinfection of other types of HBV. (1) Virus factors: In most cases there is no change in the \u03b1 determinant that could explain the lack of HBsAg detection In this study, we found that patients with occult HBV infection are less likely to develop cirrhosis and had better overall survival. This observation strongly supported the possible contribution of OBI to the establishment of cirrhosis and the possible direct or indirect role in the development of HCC. In patients with diagnosable/detectable low-grade HBV replication, the virus retains its pro-oncogenic properties Our study has limitations. Firstly, samples were from a single department and the size was limited. In future studies, larger sample size would be preferred in order to validate the findings shown in this study. Besides, studies should continue to functionally characterize viral mutations and the relevant viral genes.In summary, our findings suggest that the intracellular HBV DNAs, such as HBV cccDNA and tDNA are valuable biological markers for the diagnosis of occult HBV infection in HCC patitents. This would assist the clinical implementation of a more personalized therapy for viral re-activation control and improve the survival rate of OBI patients.Table S1Sequences of primers used in the study for cccDNA and tDNA.(DOC)Click here for additional data file.Table S2Correlations among HBV DNAs in the HBsAg-positive group.(DOCX)Click here for additional data file.File S1Supplemental Materials and Methods.(DOC)Click here for additional data file."} +{"text": "Cysteinyl leukotrienes (LTs) contribute to airway inflammation by interacting with type 1 and 2 cysteinyl leukotriene receptors as well as not clearly known type 3 leukotriene receptors. In contrast to LTC4 and LTD4, LTE4 is weak agonist at type 1 and 2 cysteinyl leukotriene receptors. Although LTE4 plays a key role in airway inflammation, our understanding of its potential receptor remains unclear. We investigated the effects of clopidogrel in the development of allergic airway inflammation using mouse asthma model and eosinophil cell-line.BALB/c mice were sensitized by intraperitoneal injection of ovalbumin (OVA) on days 0 and 14, followed by 3 nebulized OVA challenges on days 28-30. On each challenge day, 30mg/kg clopidogrel was administered through intragastric administration 30 minutes before challenge. 48 hours after OVA challenge, mice were assessed for airway hyperresponsiveness (AHR), cell composition and cytokine levels in bronchoalveolar lavage (BAL) fluid. Human eosinophil EOL-1 cells were treated with LTE4 with or without clopidogrel, and intracellular and extracellular eosinophil cationic protein (ECP) expression were investigated by Western blot and ELISA, respectively. Finally, CC ligand 5 (CCL5) levels in BAL fluid were measured by ELISA. Our experiments were approved by Institutional Animal Care and Use Committee of Ajou University (IACUC #152)P<0.01, respectively). These results were associated with decreased levels of Th2 cytokines and CCL5, but not Th1 cytokine in BAL fluid. In histological analysis, the inflammatory cells in peribronchial and perivascular areas as well as mucus-containing goblet cells were also decreased in the clopidogrel administered mice compared to vehicle treated mice (P<0.01). LTE4 stimulation decreased intracellular expression of ECP but this expression was attenuated by pre-treatment of clopidogrel.The administration of clopidogrel decreased AHR and airway inflammatory cell numbers including eosinophil in BAL fluid following OVA challenge (Clopidogrel could prevent the development of AHR, airway inflammation, and cytokine production in allergen challenged mice through the inhibition of eosinophilic activation. Clopidogrel could be a novel therapeutic target for asthma treatment."} +{"text": "Intercalating age-specific DNA methylation and microbiome changes were identified, which may have significant translational relevance in the developmental origins of IBD and CRC.Epigenetic and microbiome changes during pediatric development have been implicated as important elements in the developmental origins of inflammatory bowel diseases (IBDs) including Crohn's disease (CD) and ulcerative colitis (UC), which are linked to early onset colorectal cancer (CRC). Colonic mucosal samples from 22 control children between 3.5 and 17.5 years of age were studied by Infinium HumanMethylation450 BeadChips and, in 10 cases, by 454 pyrosequencing of the bacterial The epidemiology of inflammatory bowel diseases (IBDs) strongly implicates their environmental origin . IBDs (CHigh monozygotic twin discordance rates and other epidemiologic observations highlight the importance of nongenetic (such as epigenetic) processes, which are vulnerable to environmental influences, in the etiology of IBD . The incThe potential importance for epigenetics in IBD is only recently receiving more attention . There avy) mouse [The first example for nutritional imprinting via environmental exposure was described in the viable yellow Agouti (Ay) mouse . A handfy) mouse and obsey) mouse , 15. We y) mouse . We havey) mouse . Howevery) mouse .Here, we analyzed our recently published cohort of 22 control children between 3.5 and 17.5 years of age in respep < 0.05) and at least 10% methylation difference between the oldest and youngest proband. More specifically, transverse colonic mucosal biopsies were used for DNA extraction. Genomic DNA was isolated by bead beating and Qiagen TissueLyser [https://www.promega.com/~/media/files/products%20and%20services/instruments/detection/tbs%20technical%20support%20docs/s-0041.pdf)before processing towards the microarrays. The samples that passed quality control were processed by Infinium HumanMethylation450 BeadChip Kits according to the manufacturer's recommendations through automated processes in the Core Laboratory for Translational Genomics of the Baylor College of Medicine. Arrays were imaged with BeadArray Reader using standard Illumina scanner settings. The R Bioconductor minfi package [http://www.rforge.net/IMA/snpsites.txt), 390,433 CpG probes on the array were used for subsequent analysis. Beta values at these CpG sites (according to GRCh37/hg19) were then correlated with the age of the 22 control proband. CpG sites with significant (Spearman rank test p < 0.05) correlation between DNA methylation and patient age were selected. Thereafter, only those CpG sites were included into the analysis of this report where 10% or more methylation difference between the oldest (17.5 years old) and youngest (3.5 years old) patient was present. To further increase the biological relevance of our findings we focused on genomic regions where at least 2 CpG sites met selection criteria within the same genomic region (an arbitrary cutoff of 15\u2009kb was used as the \u201csame genomic region\u201d). DNA methylation at the CpG sites within these regions was examined in the transverse colonic mucosa of pediatric UC and CD patients. IBD dependent DNA methylation difference was examined by Student's t-test at these CpG sites. Those CpG sites were highlighted where t-test p value was less than 0.05.DNA isolated from left sided colonic mucosal samples was interrogated by Infinium HumanMethylation450 BeadChip Kits . We determined beta values at each CpG site and selected those with significant age-dependent correlation and the mucosal microbiome. This comparison was performed in a subset of 10 children and adolescents from the 22 controls. The DNA isolated from these patients' biopsy samples was analyzed not only by the bead-chip kits but by 454 pyrosequencing of the bacterialp < 0.05) manner during childhood and adolescence according to our criteria . DNA methylation increased at 852 sites with the same measures (Supplementary Table 2). Thereafter, we further delineated those genomic loci where at least 2 CpG sites met selection criteria within the same genomic region (an arbitrary cutoff of 15\u2009kb was used as the \u201csame genomic region\u201d). The chance for such association to occur between 2 CpG sites is <10\u22126 (these regions are highlighted in Supplementary Tables 1-2). These genomic loci were arbitrarily defined as pediatric age-dependent differentially methylated regions or CRC . Conversely, DNA methylation at 63 DMRs increased significantly during this pediatric developmental period. Thirty-seven (37 = 58.7%) of these DMRs associated with genes, but only 6 (16.2%) of those have been implicated in either colitis or CRC .There were 621 CpG sites in somatic chromosomes where methylation decreased in an age-dependent (correlation with age ns (DMRs ). Twentyp < 0.05) differentially methylated between UC and controls (Supplementary Table 3). In the meantime, only 3 of these DMRs were significantly differentially methylated in the colonic mucosa of CD patients (Supplementary Table 4). Furthermore, 7 (70%) out of the 10 genes associated with the UC specific DMRs overlapping with age-dependent DNA methylation decline have already been observed to be differentially expressed in the colonic mucosa of adult UC patients [Importantly, we observed that average DNA methylation at 11 (47.8%) of the decreasing methylation DMRs was also significantly were differentially methylated in pediatric UC patients (Supplementary Table 3), while only 4 (6.4%) were differentially methylated in CD patients (Supplementary Table 4). Three (37.5%) of the 8 genes associated with the UC DMRs in this group have been observed to be differentially expressed in the colonic mucosa of adult UC patients, while none of the CD DMRs had such an association.p for multiple testing <10\u22127) decreased between fetal and pediatric life (809 CpGs) (Supplementary Table 7). The chance for such overlap between the two compendiums to occur randomly is <10\u22127775. Four (80%) of the associated genes with these CpG sites have been linked to either ulcerative colitis (IL4R and SAA1) or CRC (LAT2 and SAA2). In addition to shared CpG sites with decreasing methylation, we found 12 CpG sites where direct overlap between age-dependent increase in DNA methylation occurred between our compendium (852 CpGs) and that of Kraiczy et al. increased) (Supplementary Table 7). The chance for such overlap between the two compendiums to occur randomly is <10\u22122,000,000,000. Five (42%) of the associated genes with these CpG sites have been linked to either ulcerative colitis (FLT1 and ELTD1) or CRC . The shared CpG sites with age-dependent DNA methylation changes between our report and the results of Kraiczy and colleagues implicate that ~0.6\u20131% of colonic mucosal epigenetic changes take place and/or proceed during childhood (beyond 3.5 years of age). Such developmentally plastic epigenetic processes are likely the most vulnerable to postnatal environmental influences thereby providing a link between environment and colonic diseases affecting the mucosa [ IL4R [ SAA1 [To further highlight the biological relevance of the developmental DNA methylation changes identified herein, we compared our results to a recently published work on intestinal DNA methylation changes from fetal to pediatric development in humans . This ree mucosa . Importaa [ IL4R and SAA1R [ SAA1 have beep < 0.1) secondary to the low number of samples available. The abundance of Roseburia, Parabacteroides, and Bacteroides increased with age ( Streptococcus and Blautia decreased during postinfantile pediatric development (p < 0.1) with the select genera abundance. Among the genes with decreasing methylation DMRs, SLC9A3 and PON1 had the highest number of genera (3/5 = 60%) correlations ( KHDC3L (C6orf221) correlated with the greatest number of genera (4/5 = 80%) ( Roseburia (Supplementary Table 5) and Streptococcus (Supplementary Table 6) were the genera with the highest number of correlations with epigenetically plastic DMRs during childhood and adolescence.We then went on to examine age related pediatric microbiome variation in the large intestine and how that may correlate with epigenetic development in children. First, we determined if bacterial genera displayed age-specific variation with respect to abundance during childhood and adolescence. Here we relaxed the significance cutoff . Conversp = 0.0007)) and DMRs with decreasing methylation during late pediatric development. The compendium of DMRs outlined herein has functional relevance in respect to mammalian (including human) colitis and CRC based upon published mucosal gene expression and functional genomic data.We detected colonic mucosal epigenetic and metagenomic plasticity during late pediatric development, which demonstrated numerous significant interactions, highlighting the importance of both epidemiological and systems biology considerations when approaching the developmental origins of gastrointestinal diseases. We observed a remarkable link between age-dependent and IBD-specific DNA methylation variation, especially in respect to UC was detected with decreased abundance in UC patients [ SLC9A3 and commensal bacteria appears to exist, which may be relevant for modulating colitis susceptibility.An outstanding example for the interactive epigenetic-metagenomic developmental plasticity is colitis . Importa Figures . In factpatients , consist KHDC3L had the highest number of microbiome associations . In facle in UC , for exap values in regard to significance are acknowledged. In the meantime, our work is the first to delineate age-dependent epigenetic and metagenomic changes in the large intestinal mucosa during human postnatal pediatric development. The selection and focus on genomic regions with at least 2 CpG sites meeting selection criteria (\u201cDMRs\u201d) increased the significance of our findings. Our previous work utilizing similar DMR criteria [p < 0.05) postnatal methylation changes in this report and the CpG sites from Kraiczy et al. [p < 10\u22127) DNA methylation changes occurred from human fetal to pediatric development. This latter result validates the findings of both independent studies and indicates that epigenetic changes in human colonic mucosa take place and/or proceed during postnatal development, similarly as in mice [The limitations of our study linking to small sample sizes and uncorrected criteria has beencriteria in the fy et al. where hi in mice .We trust that the compendium of the developmentally plastic colonic mucosal DMRs identified herein will provide high impact targets for the biomedical field in respect to the generation of novel epigenetically and metagenomically focused preventative and therapeutic measures for both IBD and CRC.https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE32146) and GSE42921 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE42921). The bacterial 16S rRNA sequences are available at NCBI SRA (BioProject: PRJNA284397).Infinium HumanMethylation450 BeadChip data for colon mucosa samples are available through the NCBI Gene Expression Omnibus under accessions GSE32146 (These tables (1\u20137) contain the developmentally shifting, and microbiome abundance linked CpG sites, which are highlighted within this publication."} +{"text": "MCPH1, significantly associating with breast cancer susceptibility both in familial and unselected cases . A total of 21 mutation positive families were identified, of which one-third exhibited also brain tumors and/or sarcomas (P = 0.0007). Mutation carriers exhibited significant increase in genomic instability assessed by cytogenetic analysis for spontaneous chromosomal rearrangements in peripheral blood lymphocytes (P = 0.0007), suggesting an effect for MCPH1 haploinsufficiency on cancer susceptibility. Furthermore, 40% of the mutation carrier tumors exhibited loss of the wild-type allele. These findings collectively provide strong evidence for MCHP1 being a novel breast cancer susceptibility gene, which warrants further investigations in other populations.Breast cancer is strongly influenced by hereditary risk factors, a majority of which still remain unknown. Here, we performed a targeted next-generation sequencing of 796 genes implicated in DNA repair in 189 Finnish breast cancer cases with indication of hereditary disease susceptibility and focused the analysis on protein truncating mutations. A recurrent heterozygous mutation was identified in early DNA damage response gene, MCHP1 gene. The genetic data combined with the evidence of genomic instability related to identified MCPH1 mutation and also loss of the other functional gene copy in several mutation carrier tumors establish MCHP1 as a novel breast cancer susceptibility gene. This provides further tools for the clinical risk assessment of individuals with family burden of breast cancer. Our results reinforce the essential involvement of DNA damage response pathway in prevention of malignancy and indicate that parallel sequencing of the genes from this pathway provides an excellent approach for the identification of novel rare inherited mutations predisposing to this common disease.Although the contribution of hereditary susceptibility to breast cancer is well-established, the majority of predisposing factors still remain unidentified. Here, we have taken advantage of recent technical and methodological advances and performed a massive parallel sequencing of hundreds of DNA damage response genes in breast cancer cases with indication of hereditary disease susceptibility. We identify a recurrent breast cancer predisposing mutation in BRCA1 and BRCA2 along with PALB2, are all characterized by several rare, loss-of-function mutations, usually protein truncations. These heterozygous germline mutations predispose carrier individuals to breast, but to some extent also to ovarian cancer . 16/189 of the sequenced patients carried known mutations in BRCA1/2, PALB2 or TP53 .(TIF)Click here for additional data file.S2 FigMutated allele has C (blue) and wild-type allele T (red) at the site indicated by the arrow.(TIF)Click here for additional data file.S3 Fig(A) Percentage of cell survival following olaparib administration in MCF7 cells that had been transfected with the indicated siRNAs. Vertical bars represent the standard error of the mean of four independent experiments. (B) Knockdown efficiency in the transfected MCF7 cells measured by qPCR. Corresponding figures from assays using MCF10A cells (C) and (D).(TIF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file."} +{"text": "To investigate the relationship between iloprost, cigarette smoke, and Fzd9 expression, we used human samples, mouse models, and in vitro studies. Fzd9 expression was low in human lung tumors and in progressive dysplasias. In mouse models and in vitro studies, tobacco smoke carcinogens reduced expression of Fzd9 while prostacyclin maintained or increased expression. Expression of miR-31 repressed Fzd9 expression, which was abrogated by prostacyclin. We propose a model where cigarette smoke exposure increases miR-31 expression, which leads to decreased Fzd9 expression and prevents response to iloprost. When smoke is removed miR-31 is reduced, prostacyclin can increase Fzd9 expression, and progression of dysplasia is inhibited. Fzd9 and miR-31 are candidate biomarkers for precision application of iloprost and monitoring of treatment progress. As we continue to investigate the mechanisms of prostacyclin chemoprevention and identify biomarkers for its use, we will facilitate clinical trials and speed implementation of this valuable prevention approach.Half of lung cancers are diagnosed in former smokers, leading to a significant treatment burden in this population. Chemoprevention in former smokers using the prostacyclin analogue iloprost reduces endobronchial dysplasia, a premalignant lung lesion. Iloprost requires the presence of the WNT receptor Frizzled 9 (Fzd9) for inhibition of transformed growth Lung cancer continues to be the leading cause of cancer death in the United States, with most cancers diagnosed in former smokers. While avoidance of tobacco abuse is clearly the strongest deterrent of lung cancer, chemoprevention in former smokers may be a more effective intervention than treatment of established lung cancers. Former smokers treated with iloprost in a phase II clinical trial had improved endobronchial dysplasia (a precancerous lung lesion)35in vitroAs a known activator of PPAR\u03b3, Frizzled 9 (Fzd9) was proposed as an alternative receptor for prostacyclin in the lung. Fzd9 is a G-protein coupled transmembrane receptor that is required for the tumor suppressive activity of the Wnt7a signaling pathway in the lung. Binding to Fzd9 by its endogenous ligand Wnt7a leads to increased PPAR\u03b3 activity and maintenance of a normal lung epithelium7To determine whether Fzd9 expression changes in human lung cancer, we measured Fzd9 mRNA levels in matched normal and tumor lung tissue. Seventy-five percent (13/17 pairs) of patient samples had higher Fzd9 expression in uninvolved lung tissue compared to tumor tissue. Higher Fzd9 in normal tissues resulted in normal to tumor Fzd9 expression ratios greater than 1 (positive with log(2)Y transformation), while higher Fzd9 in the tumor tissue led to normal to tumor ratios of less than 1 (negative with log(2)Y transformation) . We prev3. In whole lungs from this model, smoke exposed mice had decreased expression of Fzd9 compared to unexposed controls , and iloprost in order to elucidate mechanisms involved in prostacyclin chemoprevention. W measured Fzd9 mRNA by qPCR at multiple CSC and iloprost exposure time points to investigate the relationship between Fzd9, cigarette smoke, and prostacyclin. The first culture of HBEC was split into three plates for CSC or iloprost and three plates for control. Plates were then carried and exposed continuously for the indicated time points. In HBEC, Fzd9 expression decreased below baseline at all times points after CSC exposure gene expression occur early in the conversion of normal epithelium in smoke exposed lung, so we used EMT gene expression as a surrogate for dysplasia measurement in our HBEC 20 week modelin vitro after exposure to iloprost. The array was focused on cancer stem cell miRNA, which may affect the earliest changes in lung epithelial cells caused by carcinogen exposure and reversed by iloprost treatment. HBEC cells were exposed to 24 weeks of CSC followed by 4 weeks DMSO or 24 weeks of CSC followed by 4 weeks of iloprost. In these cells, expression of 23 miRNA was decreased with previous smoke exposure and iloprost compared to previous smoke exposure alone and cigarette smoke. Increased prostacyclin in these models leads to increase or maintenance of Fzd9 expression, supporting a relationship between prostacyclin and Fzd9 expression in the context of lung cancer development. If the mouse models parallel the human experience, Fzd9 expression will be lower in cigarette smoke-exposed populations at high risk for lung cancer. We propose a model where cigarette smoke exposure decreases Fzd9 expression, in part by increasing miR-31 and preventing normal signaling between prostacyclin and Fzd9. When cigarette smoke exposure is removed, prostacyclin reduces miR-31 expression and Fzd9 expression begins to return for endpoint analysis in patients. Data from this study has provided valuable preclinical data supporting clinical implementation of lung cancer chemoprevention. Our group recently began a Phase I trial using inhaled iloprost. The oral iloprost trial has and this trial will have specimens available for validation of Fzd9 as a prediction and response marker. This work demonstrates the value of \u201cbedside to bench\u201d translational research, where clinical data motivates biomarker and mechanistic studies that have immediate clinical relevance.We demonstrated that exposure to cigarette smoke represses Fzd9 expression and since Fzd9 is required for iloprost activation of PPAR\u03b3, this could explain the lack of response in current smokers in the oral iloprost clinical chemoprevention trial. If Fz9d expression increases with smoking cessation, this could explain the range of response seen in former smokers in the trial, which may be dependent on quit time. Our 2 incubator and passaged twice per week. Primary dysplastic lung cell cultures isolated from biopsies at the University of Colorado Cancer Center were grown in Bronchial Epithelial Basal Media (Lonza) at 37\u2009\u00b0C in a humidified 5% CO2 incubator. Morphology of cells lines was verified twice weekly. Cigarette smoke condensate (CSC) was applied at 5\u2009ug/mL concentration and Iloprost at 10\u2009uM in cell growth media. Cells were allowed 24\u2009hours for recovery after plating before drug or carcinogen application. HBEC with CSC and iloprost exposure were carried in triplicate as individual cell lines with twice a week passaging for 1\u201324 weeks. DMSO was used for a vehicle control. All long-term HBEC cultures were grown and handles in a dedicated incubator and biosafety cabinet.Human bronchial epithelial cells (HBEC) (A gift from Dr. John Minna) were cultured in Keratinocyte Serum Free Medium (GIBCO) at 37\u2009\u00b0C in a humidified 5% COThe University of Colorado Cancer Center SPORE in Lung Cancer Tissue Bank and Biomarkers Core (Tissue Bank) provides sets of publicly available, deidentified samples for small studies. The Colorado Multiple Institutional Review Board approves all the human experimental protocols that generate specimens for the Tissue Bank and collects written, informed consent from all subjects. RNA from 17 pairs of deidentified matched human normal lung and NSCLC tumor tissues were obtained from the Tissue Bank after application to the Tissue Use Committee.3\u2009All procedures were approved by the Denver Veterans Administration Medical Center institutional animal care and use committee. Mouse tissues used in this study were previously generated and frozen in RNALater (Qiagen). For urethane studies, wild type AJ or wildtype and prostacyclin synthase transgenic (PGIStg) FVB mice, one 1\u2009mg/g urethane i.p. injection was given and mice were sacrificed 20 weeks laterRNA was extracted from HBEC and dysplastic cells using the AllPrep Universal kit (Qiagen) and from mouse tissue using the AllPrep DNA/RNA kit (Qiagen) on the Qiacube automated processor (Qiagen). mRNA was reverse transcribed using the ABI HC cDNA kit (Thermo Fisher Scientific) and miRNA using the miScript II RT Kit (Qiagen). qPCR primers included: hu and ms Fzd9, hu and ms GAPDH, ms 18sRNA, hu Snail, hu Ecadherin, hu Vimentin, hu Crumbs3, and hu Zona Occludens-1 mRNA (Bio-Rad) and for hsa-miR-31 and RNU6 miRNA (Qiagen). qPCR was conducted using standard protocols for Sso Advanced SYBR Green Master Mix (Bio-Rad) or the miScript SYBR Green PCR Kit (Qiagen) on a CFX96 Touch (Bio-Rad). Genes for normalization of mouse model PCR were determined by screening 8 genes with the Mouse Housekeeping Genes PCR Array . All PCR reactions were conducted in triplicate.For initial miRNA screening, HBEC cells were treated with 24 weeks CSC followed by 4 weeks with DMSO or iloprost. Total RNA was extracted and reverse transcribed with a miScript II RT kit (Qiagen). miRNA expression was screened using a miScript miRNA PCR Array focused on cancer stem cells . The array profiled expression of 84 miRNAs and included a set of six normalization controls as well as controls to assess RNA recovery, reverse transcription performance, and PCR performance. Results were analyzed through the SABiosciences data analysis portal.HBEC cells were cultured with iloprost for 6 weeks. Cells were then transfected with hsa-miR-31 mimic (2.5\u2009nM) (Qiagen) or a negative control (2.5\u2009nM) and 1.1\u2009ul TransIT-X2 (Mirus Bio) per well in 24-well plates. After 48\u2009hours, total RNA was extracted for qPCR analysis. Transfections were done in duplicate.For single comparisons, a one-sided t-test was used to calculate p-values. For multiple comparisons, a planned comparison approach was usedHow to cite this article: Tennis, M. A. et al. Prostacyclin reverses the cigarette smoke-induced decrease in pulmonary Frizzled 9 expression through miR-31. Sci. Rep. 6, 28519; doi: 10.1038/srep28519 (2016)."} +{"text": "PTEN) is a well established tumor suppressor gene. Recently, increasing studies investigated the association between PTEN IVS4 polymorphism (rs3830675) and risk of various types of cancer. However, the results from the individual studies were controversial. The aim of this meta-analysis was to elucidate whether PTEN IVS4 polymorphism was associated with cancer risk.Phosphatase and tensin homolog were systematically searched to identify potentially eligible literatures. Odds ratios (OR) and their 95% confidence interval (CI) were used to assess the strength of association between PTEN IVS4 (\u2212/\u2212) genotype were significantly associated with increased risk of cancer and subgroup of digestive tract cancer compared with (+/+) genotype. The allele analysis revealed that (\u2212) allele was significantly associated with increased risk of cancer and subgroup of digestive tract cancer compared with (+) allele. No significant association was observed between PTEN IVS4 (+/\u2212) genotype and risk of cancer.A total of seven case-control studies were finally included in this meta-analysis. The pooled analysis suggested that individuals with PTEN IVS4 (\u2212/\u2212) genotype was significantly associated with increased risk of cancer especially for digestive tract cancer compared with (+/+) genotype. The (\u2212) allele of PTEN IVS4 (rs3830675) polymorphism was significantly associated with increased risk of cancer especially for digestive tract cancer compared with (+) allele. The recessive effect model and dominant effect model also demonstrated significant association between PTEN IVS4 (rs3830675) polymorphism and increased cancer risk especially for digestive tract cancer. Further large-scale and well-designed studies regarding different ethnicities are still required to confirm the results of our meta-analysis. The global burden of cancer continues to increase largely with approximately 12.7 million cancer cases and 7.6 million cancer deaths each year worldwide PTEN), a well established tumor suppressor gene, is mapped to chromosome 10q23.3. PTEN gene spans 105 kb including nine exons and eight introns. The 403 amino-acid PTEN protein is a dual phosphatase, acting at both serine-threonine and tyrosine sites PTEN mutations have been widely reported in various types of cancer including prostate cancer, breast cancer, endometrial cancer and so on PTEN gene. PTEN polymorphisms have been reported to be involved in multiple cancers, such as breast cancer Phosphatase and tensin homolog (PTEN IVS4 polymorphism (rs3830675) with ATCTT insertion at 109 bp downstream of exon 4 in intron 4 was one of the common PTEN polymorphisms. Most recently, increasing studies investigated the association between PTEN IVS4 polymorphism (rs3830675) and risk of various types of cancer including breast cancer PTEN IVS4 (\u2212/\u2212) genotype was associated with increased risk of colorectal cancer PTEN IVS4 polymorphism and susceptibility to prostate cancer PTEN IVS4 polymorphism (rs3830675) with susceptibility to cancer. To explore whether PTEN IVS4 polymorphism (rs3830675) was associated with risks of cancer and specific cancer subtypes, we performed a meta-analysis on the association between PTEN IVS4 polymorphism (rs3830675) and cancer risk in the present study.Until now, no meta-analysis has been performed to investigate the association of Electronic databases of Web of Science, PubMed and Chinese National Knowledge Infrastructure (CNKI) were systematically searched using different combinations of the search terms including \u201cphosphatase and tensin homolog/PTEN\u201d, \u201cpolymorphism/mutation/variant\u201d and \u201ccancer/neoplasm/malignancy\u201d. The corresponding Chinese search terms were used when searching Chinese database. References cited in each included literatures were further searched manually to identify potential available studies. If the information provided in the literature was not sufficiently clear, the author was contacted for additional raw data. When overlapping data exists, only the study with more complete information was adopted. The last search date was November 12th, 2013.PTEN gene IVS4 polymorphism (rs3830675) and risk of cancer; studies published in English or Chinese; studies with sufficient raw data for estimating odds ratios (OR) and their 95% confidence interval (CI). The main reasons for exclusion were: duplicate data; reviews; not relevant to cancer or specific polymorphism; animal experiments; not case-control designed; no raw data even after contacting the author.All the included studies must meet the following criteria: case-control study; studies concerning the association between PTEN gene IVS4 polymorphism, source of controls. The conflict was resolved after discussion and consensus was finally reached on all of the extracted data.Two authors (Liping Sun and Jingwei Liu) independently extracted the data from the included studies. The following data were extracted from each individual study: first author, publication year, ethnicity of the studied population, cancer type, numbers of each genotype in cases and controls, genotyping methods for PTEN gene polymorphism and cancer risk. P value<0.05 was regarded as statistically significant. Heterogeneity was assessed by I-squared (I2) value and using Q statistic (P<0.10 suggests significant heterogeneity between studies) All the statistical analysis was performed by Stata software . ORs and their 95%CI were used to assess the strength of association between This meta-analysis was organized based on PRISMA statement (PRISMA Checklist). A total of 492 literatures were obtained from electronic databases after duplicates removal. After reviewing the titles and abstracts, 468 articles were excluded mainly due to no relevance, reviews, animal experiments or not about cancer. Subsequently, the left 24 publications were further evaluated for eligibility. Seventeen literatures were removed because of not in English or Chinese, no raw data, not case-control designed or not concerning IVS4 polymorphism. Finally, seven full-text articles with eligibility were included in the present meta-analysis. The detailed flow chart of study selection was shown in PTEN IVS4 polymorphism in each study was all detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The studied cancer types of the publications consisted of breast cancer, colorectal cancer, gastric cancer, esophageal cancer and prostate cancer. Cancer types including esophageal cancer, gastric cancer and colorectal cancer were integrated as digestive tract cancer when performing subgroup analysis based on cancer type.The baseline characteristics of the studies included in this meta-analysis were summarized in PTEN IVS4 (\u2212/\u2212) genotype were significantly associated with increased risk of cancer genotype was observed to be associated with increased risk of digestive tract cancer but no significant association was found with breast cancer or prostate cancer . As for different ethnicities, carriers of PTEN IVS4 (\u2212/\u2212) genotype were significantly associated with increased risk of cancer in Turkish and Chinese, but not in American population. In the subgroup analysis based on source of control, increased cancer risk was observed in population-based subgroup rather than hospital-based subgroup.The results of the quantitative synthesis of the data were summarized in PTEN IVS4 (+/\u2212) genotype were not significantly associated with risk of cancer genotype was significantly related with increased risk of digestive tract cancer ; while no such association was detected in subgroups of breast cancer or prostate cancer. In addition, no significant association was observed in different subgroup based on ethnicities. It was worth noting that subgroup analysis of population-based (PB) and hospital-based (HB) demonstrated controversial outcomes , which suggested that the selection of the controls might influence the result of the relation between PTEN IVS4 (+/\u2212) genotype and cancer risk.Individuals with PTEN IVS4 (rs3830675) polymorphism and increased cancer risk (\u2212/\u2212 vs. (\u2212/+ and +/+): OR\u200a=\u200a1.56, 95% CI\u200a=\u200a1.19\u20132.05, P\u200a=\u200a0.001, The recessive effect model and dominant effect model also demonstrated significant association between PTEN IVS4 (rs3830675) polymorphism was significantly associated with increased risk of cancer allele was observed in Turkish and Chinese populations, but no significant relation was found in American.The allele analysis revealed that (\u2212) allele of PTEN IVS4 (\u2212/\u2212) genotype with (+/+) genotype, no significant heterogeneity was observed in the overall analysis and all of the subgroup analyses. Therefore, fix-effect model was adopted. In several comparisons of PTEN IVS4 (+/\u2212) genotype with (+/+) genotype and allele analysis, random-effect model were used due to significant heterogeneity (In the comparison of ogeneity . SensitiPTEN gene IVS4 (rs3830675) polymorphism. Besides, funnel plot that qualitatively assessed the publication bias of association between PTEN IVS4 (\u2212/\u2212) genotype and cancer risk was presented in The Begg\u2019s test and Egger\u2019s test were performed to evaluate the publication bias of the studies quantitatively. The detailed information for publication bias test was shown in PTEN gene IVS4 (rs3830675) polymorphism with cancer risk turn out to be controversial PTEN IVS4 polymorphism in carcinogenesis. By performing the current meta-analysis, we suggested that PTEN IVS4 (\u2212/\u2212) genotype and the (\u2212) allele were significantly associated with increased risk of cancer especially for digestive tract cancer, respectively. However, no significant association was observed between PTEN IVS4 (+/\u2212) genotype and risk of cancer.Results of previous studies concerning the relationship of PTEN gene in carcinogenesis has attracted special interest. Somatic mutations have been detected in a variety of cancer including breast, prostate, melanoma and endometrial PTEN predispose carriers to develop Cowden\u2019s disease and Bannayan-Zonana syndrome PTEN gene and risk of cancer has been investigated in various types of cancer. One of the most commonly studied polymorphism was PTEN gene IVS4 (rs3830675) polymorphism in intron 4. As individual studies demonstrated inconsistent results, we performed the present meta-analysis to elucidate the exact role of PTEN gene IVS4 (rs3830675) polymorphism in carcinogenesis.After its first identification as a tumor suppressor gene in 1997, the role of PTEN IVS4 (\u2212/\u2212) genotype was significantly associated with increased risk of cancer (OR\u200a=\u200a1.45) and subgroup of digestive tract cancer (OR\u200a=\u200a1.67) compared with (+/+) genotype; no significant association was observed between (\u2212/\u2212) genotype and risks of breast cancer or prostate cancer. The recessive effect model and dominant effect model also demonstrated significant association between PTEN IVS4 (rs3830675) polymorphism and increased cancer risk (\u2212/\u2212 vs. (\u2212/+ and +/+): OR\u200a=\u200a1.56; (\u2212/+ and \u2212/\u2212) vs. (+/+): OR\u200a=\u200a1.26) especially for digestive tract cancer. Similarly, (\u2212) allele of PTEN IVS4 (rs3830675) polymorphism was also significantly associated with increased risk of cancer (OR\u200a=\u200a1.30) and digestive tract cancer (OR\u200a=\u200a1.42) compared with (+) allele; no such significant association was found in subgroups of breast cancer or prostate cancer. Different cancer has its distinct mechanisms of initiation and progression, the obvious different outcome of diverse cancer subtypes in this meta-analysis indicated that PTEN IVS4 (rs3830675) polymorphism might confer altered risks to various types of cancer. The consistent association between PTEN IVS4 (rs3830675) polymorphism and risk of digestive tract cancer in different genetic models suggested that PTEN might have specific role in the carcinogenesis of digestive tract cancer, which requires further studies to explore. As for different ethnicities, PTEN IVS4 (\u2212/\u2212) genotype and (\u2212) allele were associated with increased risk of cancer in both Turkish and Chinese but not in American. Although different associations were observed in Asian and Caucasian, conclusion based on different ethnicities should be drawn carefully due to the limited study number for Caucasians. In addition, heterozygous (+/\u2212) genotype was not significantly associated with altered cancer risk according to the pooled estimate in this meta-analysis.By performing the current meta-analysis, we found that PTEN as a tumor suppressor has been firmly established. PTEN antagonizes the phosphoinositol-3-kinase/AKT signaling pathway and suppresses cell survival as well as cell proliferation, thereby safeguards important cellular machineries against tumorigenesis PTEN polymorphisms mainly focused on the exon region of PTEN gene. Quite recently, the intron polymorphism of PTEN drew increasing attention. Although introns were originally believed to be nonfunctional because they do not code for proteins, it has been suggested that some of these sequences possess crucial functions PTEN IVS4 (rs3830675) polymorphism may lead to a splicing error or may be by linkage disequilibrium with another locus to affect the expression and function of the PTEN. The alternation of PTEN expression would inevitably change the role of PTEN in maintaining genome stability, and loss of function of this tumor suppressor might therefore leads to carcinogenesis. Although the above-mentioned possible mechanism might partially explain the observed association between PTEN IVS4 (rs3830675) polymorphism and cancer susceptibilities, rare functional study has been carried out and the exact mechanism remain largely elusive. Future functional studies are still needed to clarify the exact mechanism of the role of PTEN IVS4 (rs3830675) polymorphism in carcinogenesis.The role of Several limitations should be acknowledged in this meta-analysis when interpreting the results. First, the sample size of this meta-analysis was not sufficiently large especially for the subgroup analyses. Second, only the studies published in English or Chinese were included in this meta-analysis, which might cause publication bias. Third, the ethnicities of most studies were Asian populations with only one study carried out in Caucasian, which deserves further confirmations in other ethnicities. Finally, because other important data such as sex, age, family history and environment risk factors were not available, we could not obtained results with adjustments by other co-variables.PTEN IVS4 (\u2212/\u2212) genotype was significantly associated with increased risk of cancer especially for digestive tract cancer compared with (+/+) genotype. The (\u2212) allele of PTEN IVS4 (rs3830675) polymorphism was significantly associated with increased risk of cancer especially for digestive tract cancer compared with (+) allele. The recessive effect model and dominant effect model also demonstrated significant association between PTEN IVS4 (rs3830675) polymorphism and increased cancer risk especially for digestive tract cancer. Further large-scale and well-designed studies regarding different ethnicities are still required to confirm the results of our meta-analysis.To be concluded, this meta-analysis indicated that Figure S1Funnel plot for studies of association between PTEN IVS4 (rs3830675) polymorphism and cancer risk (\u2212/\u2212 vs. +/+).(TIF)Click here for additional data file.Figure S2Funnel plot for studies of association between PTEN IVS4 (rs3830675) polymorphism and cancer risk (\u2212/\u2212 vs. [\u2212/+ and +/+]).(TIF)Click here for additional data file.Figure S3Funnel plot for studies of association between PTEN IVS4 (rs3830675) polymorphism and cancer risk ([\u2212/\u2212 and \u2212/+] vs. +/+).(TIF)Click here for additional data file.Checklist S1PRISMA checklist.(DOC)Click here for additional data file."} +{"text": "Management of cardiac surgery patients is a very standardized procedure in respective local institutions. Yet only very limited evidence exists concerning optimal indication, safety and efficacy of hemodynamic monitoring catecholamine and fluid therapy.Between April and May 2013, all 81 German anaesthesia departments involved in cardiac surgery care were asked to participate in a questionnaire addressing the institutional specific current practice in hemodynamic monitoring, catecholamine and volume therapy.51 (63%) questionnaires were completed and returned. All participating centers used basic hemodynamic monitoring , supplemented by transesophageal echocardiography. Pulmonary arterial catheter and calibrated trend monitoring devices were also routinely available. In contrast, non-calibrated trend monitoring and esophageal doppler ultrasound devices were not commonly in use. Cerebral oximetry is increasingly emerging, but lacks clear indications. The majority of patients undergoing cardiac surgery, especially in university hospitals, required catecholamines during perioperative care, In case of low cardiac output syndrome, dobutamine (32%), epinephrine (30%) or phosphodiesterase inhibitors (8%) were first choice. In case of hypotension following vasoplegia, norepinephrine (96%) represented the most common catecholamine. 88% of the participating centers reported regular use of colloid fluids, with hydroxyethyl starches (HES) being first choice (64%).Choice of hemodynamic monitoring is homogenous throughout German centers treating cardiac surgery patients. Norepinephrine is the first line catecholamine in cases of decrease in peripheral vascular resistance. However, catecholamine choice for low cardiac output syndrome varies considerably. HES was the primary colloid used for fluid resuscitation. After conduct of this survey, HES use was restricted by European regulatory authorities in critically ill patients and should only be considered as second-line fluid in surgical patients without renal impairment or severe coagulopathy. Large clinical studies addressing catecholamine and fluid therapy in cardiac surgery patients are lacking. An estimated 100.000 cardiac surgical procedures are performed each year in Germany Despite the fact that the intraoperative anesthesiological approach to cardiac surgical procedures are usually standardized within the setting of the respective institutions, very little to no clinical data is available concerning appropriate intraoperative hemodynamic monitoring, vital parameter goals, and catecholamine and volume therapy in cardiac surgery patients In 2005 the German Society for Thoracic and Cardiovascular Surgery (DGTH) and the German Society for Anaesthesiology and Intensive Care Medicine (DGAI) initiated and then updated S3 guidelines for postoperative intensive care in cardiac surgery patients We present the results of a postal survey evaluating the current intraoperative practice regarding hemodynamic monitoring, catecholamine and volume therapy at German cardiothoracic anaesthesia centers. The results of this survey could serve as a basis for the development of guidelines for the intraoperative care of cardiac surgery patients.The study was approved by Jena University Ethics Committee which waived informed consent because of the anonymous nature of the study.A postal questionnaire was sent by the DGAI to the department heads of the 81 institutions that have a cardiac surgery department in Germany. There was a covering letter explaining the aims of the study and a stamped addressed return envelope for return postage. The letters were sent to the hospitals in the period between 01 April 2013 and 31 May 2013. All letters were delivered by mail, and no letters were returned because of an invalid address. Due to the fact that the acquisition of the data was performed anonymously and the questionnaires were collected by the society, no estimate of survey characteristics for nonrespondents and respondents can be made to assess the potential nonresponse bias.The questionnaire consisted of 23 questions covering four major areas: a) structural data regarding hospital structure and patient care, b) standard procedures of hemodynamic monitoring as well as implementation of advanced regional perfusion monitoring devices, c) use of first- and second line vasoactive agents or inotropic drugs in hypotension following low cardiac output syndrome or vasoplegia and d) different volume replacement strategies, with a special focus on colloidal fluids and crystalloids. Frequencies in the use of different monitoring devices or volume replacement were categorized on a one to five graded Likert scales ranging from one - always present/is always true to - five - very rare/not present. The questionnaire itself is provided in Evaluation of the questionnaires were performed anonymously after having collected all the returned sheets. Due to the study design all missing values represented missing answers. Categorical data were assessed and depicted by frequencies. Values graded on Likert scales were analyzed by descriptives calculated with IBM SPSS statistics Version 21.51 of the 81 (63%) institutions with a cardiac surgery department in Germany answered and returned the questionnaire. Of these, 50 questionnaires were eligible for further evaluation. One questionnaire from a pediatric cardiac surgery department was excluded from further analysis, because of specific pediatric operative procedures and pathophysiology that could possibly influence fluid administration and catecholamine use.17 (34%) centers conducted 1000 to 1500 cardiac operations per year, 11 (22%) centers between 1500 and 2000 and 7 (14%) more than 2000 operative procedures per year. 8 (16%) and 6 (12%) centers performed between 750 and 1000 or less than 750 cardiac operative procedures per year, respectively (All centers participating in this survey (n\u200a=\u200a50 (100%)) used intraoperative transesophageal echocardiography (TEE) to monitor macrohemodynamic parameters in addition to bedside basic monitoring ) reported the use of basic monitoring . With a median value of 1 measurement of central venous pressure (CVP) was also a common parameter of intraoperative macrohemodynamic monitoring. Moreover, TEE was frequently used with a median value of 2 , followed by PA catheterization and calibrated trend monitoring systems . The actual application of uncalibrated trend monitoring devices , transesophageal doppler sonography or other devices (i.e. TCD or LAP) only played a minor role in measuring intraoperative hemodynamics (36 (72%) of all returned questionnaires reported that 80\u2013100% of patients received catecholamines within the perioperative period. Another 7 (14%) centers reported 60\u201380% and 4 (8%) of 40\u201360% catecholamine dependency in patients during perioperative care. Only 2 (4%) centers reported that less than 35% of patients needed catecholamines (Standard operating procedures (SOP) for intraoperative catecholamine use were available in 23 (46%) of the participating centers, while 25 (50%) had no SOP for intraoperative catecholamine use. In the postoperative period 18 (36%) centers applied an existing SOP for catecholamine administration, while 22 (44%) centers had no such SOP. In 20% the availability of a SOP for postoperative catecholamine use remained unknown . Nevertheless, 62% of the participating centers applied fluid challenges and/or used catecholamines in case of decreasing mean arterial blood pressure (MAP) or systolic blood pressure (systBP).Seven centers reported the use of intraoperative fluid challenges or catecholamines in case of decreasing MAP below 60 mmHg or systBP below 80 mmHg (n\u200a=\u200a2). In cases of absent severe comorbidity lower median MAP limit (n\u200a=\u200a18) or systBP (n\u200a=\u200a6) was 57.5 mmHg or 90 mmHg , respectively. On the other hand, when severe comorbidity was present, the lower median MAP limit (n\u200a=\u200a18) or systBP (n\u200a=\u200a5) increased to 70 mmHg or 100 mmHg , respectively (In case of hypotension caused by low cardiac output the most commonly used catecholamine was dobutamine (n\u200a=\u200a16 (32%)), followed by epinephrine (n\u200a=\u200a15 (30%)) and phosphodiesterase inhibitors (PDE-inhibitors (n\u200a=\u200a4 (8%)). All other substances were rarely used figure 448 (96%) of all centers used norepinephrine as the first line catecholamine in patients with hypotension caused by vasoplegia, followed by ephedrine (n\u200a=\u200a1 (2%) or phenylephrine+norepinephrine , right ventricular filling pressures (i.e. CVP) or TEE were predominantly used for monitoring of fluid administration. Urine output , PA occlusion pressure and trend monitoring systems (i.e. PICCO or Vigileo) were more seldom used figure 688% of the participating centers reported regular colloid use in their patients , while 32% (n\u200a=\u200a16) of the participating centers used HES fluids. In 10% (n\u200a=\u200a10) gelatine or albumin (4% (n\u200a=\u200a2)) was used for priming figure 858% (n\u200a=\u200a29) and 44% (n\u200a=\u200a22) of all participating centers applied a SOP for intraoperative or postoperative erythrocyte transfusion, respectively.table S1 in File S2). However, intraoperative use of calibrated trend monitoring devices were almost equally ranked on the 1 to 5 Likert scale among the different level of the participating centers (table S2 in File S2). Intraoperative regional perfusion monitoring with cerebral oximetry devices were equally distributed among all centers, whereas continuous mixed venous or central venous saturation monitoring were almost exclusively available in university hospitals (table S3 in File S2). Specialized heart centers or maximal care hospitals more frequently applied no intraoperative regional perfusion monitoring device whatsoever (table S4 in File S2).Catecholamine- and volume therapy substantially varied between the different levels of hospital care. Calibrated trend monitoring devices seemed to be more commonly available in university and maximal care hospitals in comparison to specialized heart centers (figure S1 in File S2). Interestingly, 4 (22.2)% of all participating cardiac anesthesia departments located at university hospitals reported to be influenced in their particular choice of catecholamines, e.g. by the surgeons (table S5 in File S2).The participating university hospitals reported that 80\u2013100% of all patients had some form of catecholamine therapy , Specialized heart centers and maximal care hospitals mainly performed cardiopulmonary bypass priming exclusively with crystalloids, while almost 60% of the participating university hospitals used colloidal fluids as priming solutions (table S7 in File S2). Furthermore, specialized heart centers more frequently applied standard operating procedures for intraoperative (81%) or postoperative (75%) red blood cell transfusion, while this was less common among the participating university and maximal care hospitals (table S8 in File S2).Among all participating centers, postoperative fluid administration was mainly conducted with crystalloids, while intraoperative colloidal fluid administration was much less common among university hospitals in Germany. Epinephrine (41.8%), dobutamine (30.9%) and PDE-III inhibitors (14.5%) were the most common drugs The wide range of catecholamine approaches in LCOS is certainly an indicator of the lack of sound clinical data pointing towards an ideal drug therapy. Hence, more clinical studies are needed to investigate catecholamine use in LCOS in patients undergoing cardiac surgery.When this survey was conducted, the latest S3 guideline for intensive care in cardiac surgery still recommended both crystalloids and HES or other colloids for postoperative care patients Volume replacement strategies in cardiac surgery have changed during the last years, reflecting increasing doubt about the efficacy and safety of HES solutions Limitation of our survey include (1) lack of a prospective study design due a focused survey, (2) limited spectrum of questions within a postal survey and (3) the fact that the survey is mainly based on answers by one or few members of the cardiac anesthesia team rather than the whole team involved in cardiac surgery care of the respective institutions.This questionnaire focused on basic and regional monitoring, catecholamine as well as volume therapy among patients undergoing cardiac surgery in Germany. This survey shows a highly standardized basic hemodynamic monitoring among all participating centers. Regional perfusion monitoring, especially cerebral oximetry, is used as an additional monitoring. Catecholamine therapy in the treatment of LCOS is heterogeneous and therefore further clinical research as well as the development of clinical guidelines is warranted. Momentarily synthetic colloid fluids are a common part of cardiac anesthesia procedures in Germany. In the light of potential risk factors associated with synthetic colloids further clinical research is also urgently needed.File S1Questionnaire hemodynamic monitoring, catecholamine and volume therapy in cardiac surgery patients.(DOC)Click here for additional data file.File S2Distinctions in hemodynamic monitoring, catecholamine- and volume therapy among different levels of hospital care - Tables and Figures - Table S1: Availability of devices for intraoperative macrohemodynamic control/global perfusion monitoring among different levels of hospital care. Table S2: frequency of the intraoperative use of hemodynamic monitoring devices among different levels of hospital care. Table S3: Availability of special monitoring devices for regional perfusion control or for oxygen consumption among different levels of hospital care. Table S4: Frequency of the intraoperative use of the regional perfusion monitoring devices among different levels of hospital care. Table S5: Influence of catecholamine therapy by others . Table S6: Intraoperative application of colloidal fluids among different levels of hospital care. Table S7: Priming solution for cardiopulmonary bypass among different levels of hospital care. Table S8: Presence of a standard operating procedure for perioperative transfusion of packed red blood cells among different levels of hospital care. Figure S1: Percentage of catecholamine use in hospitals of different levels of care.(DOC)Click here for additional data file."} +{"text": "Marianna Ricci and Paola Ulivi are not affiliated with #2 but with #3 Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy.The affiliations for the 10"} +{"text": "However, how dmy makes the male decision via initiating testicular differentiation has remained unknown. Here we report that autosomal gsdf serves a male sex initiator. Gene addition and deletion revealed that gsdf was necessary and sufficient for maleness via initiating testicular differentiation. We show that gsdf transcription is activated directly by dmy. These results establish the autosomal gsdf as the first male sex initiator. We propose that dmy determines maleness through activating gsdf and dmrt1 without its own participation in developmental processes of sex initiation and maintenance. gsdf may easily become a sex determiner or other autosomal genes can be recruited as new sex determiners to initiate gsdf expression. Our findings offer new insights into molecular mechanisms underlying sex development and evolution of sex-controlling genes in vertebrates.Sex is pivotal for reproduction, healthcare and evolution. In the fish medaka, the Y-chromosomal Sex development is a multi-step process including sex determination, initiation, differentiation and maintenance, culminating in the production of sperm or eggs for germline transmission. Defects in each of these steps may lead to sex abnormalities including infertility and sex reversal. Animal sex control has an impact in animal husbandry and even human healthcareOryzias latipes) is an excellent vertebrate model for sex development8sry, namely dmydmrt1ydmrt1bYdmy activates dmrt1dmrt1 mutation causes male-to-female sex reversal after the initiation of testicular differentiationdmy exerts its primary role in male decision via triggering testicular differentiation has remained unknown. Paradoxically, there are several cases where dmy is dispensable for maleness13181418The enormous diversity of genetic sex determination mechanisms is a long-standing mystery and also a major challenge for understanding sex developmentgsdf is emerging as a novel sex related factor in several distantly related fish species. This teleost-specific genegsdf is located on chromosome 12 and is predominantly expressed in the Sertoli cells and granulosa cells in mature gonadsgsdf acts as a male sex initiator downstream of dmy and renders itself a prime candidate for the searched autosomal gene essential for maleness.Recently, the gene dmygsdf\u2009Y, a Y-chromosomal copy of gsdf acting as the SD in O. luzonesisgsdf was similar to gsdf\u2009Y in function. To this end, pGsdf2Agfp expressing the Gsdf and GFP from the \u03b2-actin promoter was constructed and XY males (n\u2009=\u20092). Intercrossing between F1 heterozygotes (gsdf+/\u2212) produced 120 F2 adults and a typical Mendelian segregation of wildtype , gsdf+/\u2212 (n\u2009=\u200962) and homozygous animals or male (n\u2009=\u200937) strictly according to their genetic sex again developed according to genetic sex, whereas gsdf\u2212/\u2212 adults were invariantly female in phenotype 27. A malnslation . Upon crgsdf\u2212/\u2212) . gsdf dietic sex . Interese female ,f. The ghenotype . The Menin situ hybridization (FISH) and immunofluorescence (IF). We first focused on dmrt1 and foxl2. The former is expressed in Sertoli cells and essential for the maintenance of testicular differentiationdmrt1 riboprobe hybridized to the dmy transcripts due to sequence similaritydmrt1 or dmy transcripts were evident in the Sertoli cells surrounding spermatogonia, spermatocytes and sperm of the wildtype XY (wtXY) testis (gsdf-mutant XY (mtXY) gonad, the dmrt1 riboprobe produced a barely detectable signal testis but hard) testis . In the e signal , possiblY testis but presY testis ,c, sugget testes and pre-t testes ,f, in acY testis , and wasY testis , which iXY gonad . IF by uXY gonad . Gsdf prXY gonad , demonstgsdf disruption. Next-generation sequencing of adult gonads of from wtXY, wtXX and mtXY fish produced a total of 35073 unigenes, 25588 of which were widely expressed in gonads of the three genotypes. The wtXX ovary and wtXY testis had 27872 and 32212 unigenes, respectively. The mtXY gonad generated 32153 unigenes, indicating that it is not different from the wtXY testis in total number of expressed unigenes 28gsdf, dmy12sdgc, dmrt1 and sox9b. sdgc is linked closely to dmy and expressed predominantly in spermatogonia but weakly in early oocytesdmrt1 and sox9b are autosomal male markers, as dmrt1 is expressed in the Sertoli cell lineage and essential for the maintenance of testis differentiationsox9b is expressed in both sexes during early gonadal sex differentiationvasa was used as a germ cell marker30foxl2 and cyp19a1 were dramatically increased to a level comparable to that in the wtXX ovary, whereas testicular markers dmrt1 and sox9b were concurrently decreased obviously in a male gonad are entering into mitotic arrest and thus dramatically fewer than in its female counterpart, where germ cells continue propagation and enter into meiosis29gsdf+/\u2212 male was crossed to a Vg female homozygous for transgene Olvasa-gfp that expresses GFP specifically in germ cells, and embryos with GFP-labeled PGCs from the F3 generation were used for PGC observation (gsdf+/\u2212 (n\u2009=\u20099) and gsdf\u2212/\u2212 (n\u2009=\u20098) embryos (gsdf+/\u2212 (n\u2009=\u20096) embryos in several insectsdmrt1 or its paralog can be repeatedly recruited as the SD in a wide variety of organisms including medakaCynoglossus semilaevis)gsdf as a male sex initiator has a direct and important implication on the evolution of SDs as well as sex chromosomes. It is generally accepted that autosomes bearing genes capable of exerting a male determining function may evolve into Y chromosomes that will compete and possibly replace the present Y chromosomegsdf may serve as a core in understanding the evolution of SDs and cascades . The medaka strain gsdf and gfp was constructed, which predicts a nascent fusion protein that is self-cleaved by 2A sequence into Gsdf and GFP. Vectors pZN1gsdf and pZN2gsdf expressing ZFNs for targeting the medaka gsdf gene were described previouslydmy coding sequence with BamHI and EcoRI into the pCS2cherryHis backbone. Plasmids for synthesizing riboprobes were constructed by TA-cloning of PCR products into pGEM-T Easy vectors (Promega). Recombinant plasmids were sequenced on the 3130XL Sequencer (Applied Biosystems). Sequence analyses were run on Vector NTI and DNAman software.Plasmid pGsdf2Agfp expressing a chimeric mRNA between medaka Microinjection of DNA and RNA into 1-cell stage embryos was done as previously describeddmy and dmrt1 transcripts as described previouslyPhenotypic sex was judged based on the secondary sex characteristics, especially the shapes of the dorsal and anal fins in medaka adultsIsolation of genomic DNA and total RNA were done as described9oC for 30 s and 60\u2009\u00b0C for 1 min. Droplets were read by QX200 Droplet Reader and analyzed with the QuantaSoft software that determines concentration of target cDNA as copies per microliter (copies/\u03bcl) from the fraction of positive droplets using Poisson statistics\u03b2-actin.RNA quantification was performed on the QX200 automated Droplet Digital PCR system . Briefly, cDNA templates were first mixed with 2\u00d7 QX200 ddPCR EvaGreen Supermix to 20-\u03bcl volume. The mixture was transferred to a 96-well plate (Eppendorf) and subjected to oil droplets generation on the automated generator . PCR in oil droplets was run for 40 cycles of 94\u2009in situ hybridization were performed as described previouslyvasa, foxl2 probes were labeled with fluorescein isothiocyanate (FITC) and gsdf, dmrt1 probes with digoxigenin (DIG) . Horseradish peroxidase-conjugated anti-FITC and anti-DIG antibodies were used for signal detection. The TSA Plus Fluorescein/TMR system was used to amplify the fluorescence signals.Riboprobe synthesis, cryosectioning and fluorescence Immunostaining and nuclear staining with DAPI on gonadal cryosections were done as describedhttp://rsat.ulb.ac.be/rsat/, last accessed March 15, 2015). Testis samples (20\u2009mg) from dmrt1bY:GFP transgenic fish and GFP antibody (Upstate) were used for in vivo chromatin immunoprecipitation (ChIP) by using the EpiQuik Tissue Chromatin Immunoprecipitation kit (Epigentek)1616Binding sites for transcription factors were predicted using the matrixTransIT-X2 Dynamic Delivery System (Mirus). Cells were transfected at a density of ~80% confluence and propagated or analyzed at 24\u201372\u2009h post transfection.Medaka embryonic stem cell line MES1 and spermatogonial cell line SG3 were maintained as described8Adult fish were observed with stereomicroscope and photographed with the Evolution VF digital camera (MediaCybernetics) and digital camera SELP1650 (SONY). Slides were observed and photographed with the LSM 510 Meta confocal microscope (ZEISS). Cultured cells were observed with Axiovert upright microscope (Zeiss) and photographed with the AxioCam digital camera (Zeiss).http://www.ensembl.org/index.html) by using the reads mapper Tophat2 . Aligned sequences were assembled into transcripts by using Cufflinks . Cuffmerge (http://cole-trapnell-lab.github.io/cufflinks/cuffmerge/) was used to merge assemblies into a master transcriptome for analyzing differentially expressed genes (DEGs). Htseq-countRNA samples from adult gonads were sequenced as 2\u2009\u00d7\u2009100-bp paired-end reads on the HiSeq 2000 platform (Illumina) by the custom service provider AITbiotech PTE LTD (Singapore). Clean reads were mapped to the medaka genome ."} +{"text": "Cold exposure alters muscular function. Muscle cooling influences the neuromuscular activation during maximal isometric voluntary contractions (MVC) and the amplitude of surface electromyography (sEMG) ,2. It alSixteen healthy adults , with a mean age of (SD) 27.0 (2.9) years; body mass 66.3 (9.8) kg; and height 169.5 (7.8) cm participated in this experimental study. The median frequency (MF) and mean power frequency (MPF) of sEMG from tibialis anterior (TA) and gastrocnemius medialis (GM) muscles during MVC in ankle planter (PF) and dorsi-flexion (DF) against a hand-held dynamometer as well as contact times on a force plate during gait before and after cooling were measured and analysed.The MF and MPF were significantly lower P < 0.01*) in both TA and GM muscle during MVC and in TA during gait trials after cooling. However, the frequency analysis for GM muscle showed no significant difference either in MF or MPF during walking on level surface .The significant time difference might be caused by the cold induced MF and MPF decrease in sEMG. Our previous investigation reported that cooling increased the sEMG amplitude and produced fatigue like responses in the leg muscles . MoreoveModerate degree and duration of cooling may affect muscle motor unit firing rates, thus shifting the sEMG spectrum to lower frequencies, therefore decreasing the leg muscle force production. The result suggests that muscle cooling can cause cold induced frequency decrease in sEMG similar to fatigue response and lead to reduced muscle performance."} +{"text": "Single nucleotide polymorphisms of Toll like receptors modify cellular immune response and induce pro-inflammatory cytokine production and therefore could be associated with enthesitis related arthritis (ERA) and/or oligoarticular and polyarticular juvenile idiopathic arthritis (JIA).To determine whether polymorphisms of TLR2 and TLR4 influence susceptibility to ERA or JIA.DNA was extracted from blood samples of 19 ERA patients, 10 patients with oligoarticular or polyarticular JIA and 40 healthy controls, all diagnosed according to ILAR criteria. Polymorphisms of the TLR2 (Arg753Gln) and TLR4 were determined using real time and multiplex PCR.All JIA patients were carriers of wild type allele for all three polymorphisms. Regarding Arg753Gln polymorphism of TLR2, only one patient with ERA (5.56%) and 2 healthy controls (5%) were carriers of heterozygous allele. There were no homozygous mutants. All ERA patients had wild type allele for Asp299Gly polymorphism of TLR4. For Thr399Ile polymorphism of TLR4, 21.05% ERA patients were heterozygous (CT variant), and none of the ERA patients was homozygous (TT variant), . In group of healthy controls, TLR4 polymorphisms Asp299Gly and Thr399Ile were in linkage disequilibrium; 2 controls were hetrozygous and 6 homozygous variant carriers for both polymorphisms, whereas linkage disequilibrium was not found among patient groups .Polymorphisms of TLR2 and TLR4 are not associated with oligoarticular/polyarticular JIA. There was also no evidence that variants of TLR are major risk factors for ERA, however, lack of susceptibility should be confirmed on larger group of patients, since four out of 19 patients were heterozygous. A study on larger cohort is currently ongoing.None declared."} +{"text": "HrCRY2 was significantly expressed in female flower buds only while HrCO had significant expression in male flowers only. Among the three male and female floral development stages (FDS), male stage II had significant expression of most of the GISD. Information on these sex-specific expressed genes will help in elucidating sex determination mechanism in seabuckthorn.Seabuckthorn is an economically important dioecious plant in which mechanism of sex determination is unknown. The study was conducted to identify seabuckthorn homologous genes involved in floral development which may have role in sex determination. Forty four putative Genes involved in sex determination (GISD) reported in model plants were shortlisted from literature survey, and twenty nine seabuckthorn homologous sequences were identified from available seabuckthorn genomic resources. Of these, 21 genes were found to differentially express in either male or female flower bud stages. Hippophae rhamnoides commonly known as seabuckthorn belongs to the family Elaeagnaceae. Seabuckthorn berries are among the most nutritious and vitamin-rich fruits found in the plant kingdom. In general, the flesh of berries contains a diverse complex of vitamins, mineral substances such as sodium salts, potassium, calcium, carbohydrates, proteins, sugars and amino acids. [o acids. , 2. Moreo acids. . Seabucko acids. . Variouso acids. . Thus tho acids. . This inFor development of superior seabuckthorn, breeding projects target both females and male cultivars . MoreoveIn dioecious plants gender determination is regulated at genetic level by X/Y chromosome system . Many moThalictrum dioicum, floral organ identity genes were differentially expressed in early development stages of male and female flowers. This led to the conclusion that regulation of these homeotic genes resulted in gender determination in this species [Humulus lupulus). Northern hybridisation in H. lupulus showed that M1 (DEFICIENS homologue) and M2 (Petunia FLORAL BINDING PROTEIN 2 homologue) transcripts were present in the early stages of floral development of both sexes, but at later stages, expression of both genes increased in male flowers and decreased in female flowers [Differences between male and female plants are primarily detected in reproductive organs, which occur through differential growth, repression or abortion of sex organs in unisexual flowers , 14. Var species . Also th flowers , 19. Mor flowers .Silene latifolia [Cucumis sativus [Salix [S. latifolia, Rumex acetosa, and Actinidia chinensis,. different spatial and temporal development stages of flower were used [APETALA 2, CLAVATA 1 and SEPTALA 3 showed differential expression among male and female flowers of plants like Z. mays, S. latifolia, A. Officinalis [The genetic control of sex determination is well-kown in several model plant systems like atifolia \u201323, Cucu sativus \u201326, Salis [Salix , 28, etcs [Salix . Thus, gs [Salix . In ordeere used \u201333. Numeicinalis \u201336, whicHippophae rhamnoides collected from Defence Institute of High Altitude Research (DIHAR), J&K, India were used in this investigation . Three different samples of floral buds for current study were collected on the basis of phenological observations on flowering of seabuckthorn in the region of study. Flower buds start developing from the month April and flowers open in the start of May to mid-May. The flower bud samples were collected in the month of April at ten days interval, starting from dormant winter bud to when buds are about to open. This is period when female and male reproductive tissues are formed in the flower buds. Flower buds were immediately frozen in liquid nitrogen and were stored at -80 \u00b0C till further use. Male and female flower bud stages were designated as Male Stage I (MST I), Male Stage II (MST II), Male Stage III (MST III) and Female Stage I (FST I), Female Stage II (FST II) and Female Stage III (FST III) respectively as shown in The flower buds of Arabidopsis which could be potential candidates for sex determination in seabuckthorn . The sequences with longest open reading frame were used for repeats, domain and protein family identification using EBI Interpro server (http://www.ebi.ac.uk/interpro/). For Phylogenetic reconstruction of potential GISD in seabuckthorn, protein sequences of known GISD characterized in model plant species were downloaded from NCBI Genbank database , with Tm of 55\u201360\u00b0C and amplicon size between 100 bp and 250 bp (http://genex.gene-quantification.info).Primers for candidate genes were designed using the Primer3 web application , CRY1 (2), FRI (2) and TFL (2) in H. rhamnoides across three developmental stages of male and female flowers of analysed . 21 GISDanalysed details HrAP1 showed female specific expression. The expression of this gene was notably 1347 fold higher in FST II as compared to MST II. On the contrary, expression of gene HrAP2 was higher in all the male flower developmental stages withthe maximum differential expression being in MST II (7.70 fold) as compared to FST II. HrLFY and HrCLV1 showed stage specific expression in male and female FDS. The expression of HrLFY was notably higher in MST I (32.16 fold) and FST II (10.15 fold) as compared to their corresponding stages. HrCLV1 was significantly expressed in MST II (9.11 fold) and FST III (4.15 fold). On the basis of this data it is concluded that expression of gene HrAP1 is female specific while that of HrAP2 is male specific. However, the expression of gene HrLFY and HrCLV1 in male and female flowers was stage dependent.As data presented in HrFIL was higher in all FDS of female flowers showed stage dependent expression pattern which was higher in FST I (12.55 fold) and MST II (6.34 fold) as compared to their corresponding stages. Thus from the data recorded it can be concluded that expression of floral organ identity gene HrSEP3 and HrYAB5 was higher in male FDS and that of HrFIL was higher in female FDS. Also the relative expression of gene HrAG was flower developmental stage dependent rather than sex of flower.Among floral organ identity genes the expression of floral organ polarity gene flowers . The difCRYPTOCHROME2 (HrCRY2) was higher across all the female FDS as compared to male FDS. The expression of this gene was 129.3 fold higher. CRYPTOCHROME1(HrCRY1) was relatively expressed higher in all male FDS with MST II and MST III showing 6.6 fold and 2.33 fold higher expression as compared to corresponding female FDS (PHYTOCHROME B (HrPHYB) was higher in all male FDS notably MST II and MST III, which showed 25 fold and 7.5 fold higher expression with respect to FST II and FST III respectively (CONSTANS (HrCO) responsible for flowering in long days was higher in all male FDS (CO (HrCOLK) showed similar pattern of expression but relative difference in a expression level was less pronounced in male and female FDS as compared to HrCO. FRIGADIA (HrFRI) and its second hoologue HrFRILK responsible for delayed flowering in absence of cold temperatures, were also found to have elevated expression in male FDS as compared to their corresponding female FDS . The relETHYLENE RESPONSE SENSOR 1 (HrERS1) and ETHYLENE RECEPTOR 1 (HrETR1) was higher in male flowers. HrERS1 suggested that these genes could play an important role in sex determination [HrAP1 (HrAP2 (AP2 plays an important role for sex determination in maize [TASSELSEED 4 (TSL4). Similarly expression of another floral meristem identity gene HrCLV1 Click here for additional data file.S2 File(TXT)Click here for additional data file.S3 File(TXT)Click here for additional data file.S4 File(TXT)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file."} +{"text": "Ldlr\u2212/\u2212 mouse monocytes rapidly accumulate cytoplasmic neutral lipid vesicles during hyperlipidaemia. Functional analysis in vivo revealed impaired monocyte chemotaxis towards peritonitis following high fat diet due to retention of monocytes in the greater omentum. In vitro assays using human monocytes confirmed neutral lipid vesicle accumulation after exposure to LDL or VLDL. Neutral lipid accumulation did not inhibit phagocytosis, endothelial adhesion, intravascular crawling and transmigration. However, lipid loading led to a migratory defect towards C5a and disruption of cytoskeletal rearrangement, including an inhibition of RHOA signaling. These data demonstrate distinct effects of hyperlipidaemia on the chemotaxis and cytoskeletal regulation of monocyte subpopulations. These data emphasise the functional consequences of blood monocyte lipid accumulation and reveal important implications for treating inflammation, infection and atherosclerosis in the context of dyslipidaemia.Blood monocytes are heterogeneous effector cells of the innate immune system. In circulation these cells are constantly in contact with lipid-rich lipoproteins, yet this interaction is poorly characterised. Our aim was to examine the functional effect of hyperlipidaemia on blood monocytes. In the Whereas the Ly6C/GR1-low (GR1low) and homologous CD16-positive CD14-low (CD16pos) monocytes are \u2018non- classical\u2019, and respond to viral and TLR7/8 cues, and have been shown to patrol the endothelium347Monocytes are a heterogeneous, key population of the mononuclear phagocyte system that fulfil a variety of innate immune functions and have independent phenotypes from their polarised macrophage descendantsThe synthesis, processing, transport and catabolism of circulating lipid species is complex and involves many different cell types and metabolic processes10131415in-vivo since the 1960s, which are most likely neutral lipid positive blood monocytes after high fat feeding181920212223Previous work on lipoprotein biology and the mononuclear phagocyte system has mainly focused the role of LDL on macrophages and dendritic cells (DC) in atherosclerosis progressionin vitro and in the hypercholesterolemic Ldlr\u2212/\u2212 mouse. Strikingly, the extravascular chemotaxis of monocytes is impaired by lipid accumulation, in part mediated by RHOA inactivation. These findings underscore the functional role of blood monocytes and suggest that dyslipidaemia associated with monocyte neutral lipid accumulation, may result in monocyte immunosuppression.We demonstrate here that monocytes accumulate cytoplasmic neutral lipid droplets in response to LDL and VLDL, which subsequently alters their cytoskeletal dynamics both All animal procedures were carried out according to the Institutional guidelines for the care and use of experimental animals and the ARRIVE guidelines. Animal studies were approved by the UK Home Office. Blood from healthy human donors was collected under institutional guidelines with informed consent approved by NRES Committee London.Ldlr\u2212/\u2212 mice were maintained on HFD or chow for 16 weeks. To induce peritonitis, mice were injected intraperitoneally (IP) with 1ml sterile 4% thioglycollate medium. After 72\u2009hours, mice were culled and the peritoneal cavity was lavaged with 10\u2009ml ice-cold PBS. Approximately 3\u2009\u00d7\u2009105 cells were stored in Tri-Reagent (Sigma Aldrich) for RNA extraction, the remainder were stained in PBS-0.5% BSA for flow cytometric analysis in a saturating concentration of anti-CD16/32 (2.4G2) using combinations of the following antibodies: anti-CD115 (AF598), anti-CD11c (N418), anti-CD45 (30-F11) (eBioscience), anti-CD11b (M1/70), anti-GR1 (RB6-8C5) anti-F4/80 (BM8) (BD Biosciences). Monocytes were defined as CD115pos CD11bpos SSCint . Mouse monocytes were purified using FACS. Surface protein expression was assessed using mouse antibodies (as above) and human HLA-DR (TU36), CD16 (3G8), CD11c (Bu15) and CD14 (M5E2) (BD Biosciences).Cells were stained for neutral lipid using LipidTox-Green (Life Technologies). To image the actin cytoskeleton cells were stained with phalloidin- AlexaFluor 488 (Life Technologies). Mounted cells were imaged using either a Zeiss AxioObserver widefield or a Leica SP5 confocal microscope with a 63\u00d7/1.4 objective. Images were analysed using Imaris software (Bitplane) or ImageJ, as indicated.Cx3cr1gfp/+ mice as previously describedPurified monocyte subsets were analysed for migration using transwells (Corning) and 2D-chemotaxis chambers (IBIDI). Adhesion and transendothelial migration (TEM) was assessed in static co-cultures using transwells and chamber slides (IBIDI). Intravital imaging of monocyte intravascular migration was assessed in high fat fed Phagocytosis was examined using fluorescence latex bead (Life Technologies) and cell death examined using Annexin V (eBioscience).Purified monocytes were treated with LDL and VLDL (100\u2009\u03bcg/ml) for 2\u2009hours and lysates collected for RHOA activation using luminescence-based G-LISA RhoA activation kit according to manufacturer instructions . CDC42 activity was assessed using colorimetric-based G-LISA kit according to manufacturer instructions and PAK activity was assessed using anti-phospho-PAK1/2/3 via Western blot.Experimental data is presented as mean +/\u2212 standard error of the mean (SEM). Populations were compared using a two-tailed Mann\u2013Whitney U test to avoid assumptions of parametric distribution. P\u2009<\u20090.05 was considered significantly different.hi and GR1low) with blood lipoproteins in the hyperlipidaemic Ldlr\u2212/\u2212 mouse with raised plasma VLDL and LDLlow monocytes . A similon (TEM) .Cx3cr1gfp/+ mice fed HFD or chow for 6 weeks. Without further genetic manipulation we were able to significantly increase both total cholesterol and LDL levels after 6 weeks HFD monocyte endothelial adhesion or transendothelial migration is not reduced after lipid treatments in vitro. Under our conditions, LDL but not VLDL increased monocyte adhesion to endothelial cells. This is in contrast to a previous report showing increased adhesion after VLDL treatmentRegardless of activation phenotype, a feature of the advanced atherosclerotic plaque is the inhibition of cellular efflux or migration from the plaque45RhoA transcripts were down regulated during HFD peritonitis; therefore we investigated whether lipid loading inhibited monocyte cytoskeletal signaling. Cytoskeletal regulation involves many proteins including the small GTPases RAC, CDC42 and RHO and is a complex and dynamic network that responds to a variety of agonistspos non-classical monocytes, which may be involved with inhibition of uropod retraction and impairment of forward cell migration\u2212/\u2212 macrophagesOur studies on the RHOA pathway were performed using a direct RHOA activator (CN03), which is based on the catalytic domain of bacterial cytotoxic necrotizing factor toxinspos monocytes, which have a migratory phenotype. This confirms Pagler et al.During atherosclerosis others have already shown that dyslipidaemia is able to perturb macrophage migration through modulation of cytoskeletal rearrangement by free cholesterol in the cell membrane3132In summary we demonstrate the effects of hyperlipidaemia on the migration and cytoskeletal regulation of monocyte subpopulations. This work extends the function of \u2018foamy monocytes\u2019 or lipophages beyond their recently demonstrated role in delivering lipid during early atherogenesisHow to cite this article: Jackson, W. D. et al. Very-low and low-density lipoproteins induce neutral lipid accumulation and impair migration in monocyte subsets. Sci. Rep. 6, 20038; doi: 10.1038/srep20038 (2016)."} +{"text": "Likelihood of sentinel node metastases can be estimated from patients\u2019 demography and primary tumor characteristics, but is precisely determined only by microscopic evaluation of excised node(s). Molecular/Genetic techniques, such as RT-PCR, Gene Expression Microarray, detection of metastasis-associated gene signatures and gene sequencing are likely to increase precision in future. Recent 10 year results of Multicenter Selective Lymphadenectomy Trial 1 (MSLT1) [Lymphatic mapping and sentinel node biopsy are widely used to stage and manage patients with intermediate and thicker primary cutaneous melanoma. (MSLT1) . Here ag"} +{"text": "ATP-binding cassette transporter A1 (ABCA1) gene, an important modulator of high-density lipoprotein cholesterol and reverse cholesterol transport, has been previously associated with plasma lipid levels, aging and CAD, but the association with CAD has yet to be replicated.Previous studies have suggested that DNA methylation contributes to coronary artery disease (CAD) risk variability. DNA hypermethylation at the ABCA1 DNA methylation levels were measured in leucocytes of 88 men using bis-pyrosequencing. We first showed that DNA methylation at the ABCA1 gene promoter locus is associated with aging and CAD occurrence in men (P\u2009<\u20090.05). The latter association is stronger among older men with CAD , who showed at least 4.7% higher ABCA1 DNA methylation levels as compared to younger men with CAD or men without CAD . Higher ABCA1 DNA methylation levels in older men were also associated with higher total cholesterol , low-density lipoprotein cholesterol and triglyceride levels . Furthermore, we showed that acetylsalicylic acid therapy is associated with 3.6% lower ABCA1 DNA methylation levels (P\u2009=\u20090.006), independent of aging and CAD status of patients.ABCA1 epigenetic profile is associated with CAD and aging, and highlights that epigenetic modifications might be a significant molecular mechanism involved in the pathophysiological processes associated with CAD. Acetylsalicylic acid treatment for CAD prevention might involve epigenetic mechanisms.This study provides new evidence that the ABCA1 gene in humans are responsible for Tangier disease (OMIM: 2054000) and familial hypoalphalipoproteinemia (OMIM: 604091)[The ATP-binding cassette transporter A1 (ABCA1) catalyzes the transfer of lipids from various tissues and cells to apolipoprotein A1 containing lipoproteins[se OMIM: 054000 anse OMIM: 054000 ania OMIM: 04091[6]ABCA1 gene promoter locus was associated with lower HDL-C levels and a previous history of CAD in familial hypercholesterolemia (FH)). The first exon of ABCA1 is in red and bold type. The epigenotyped region is shown in green. Arrows indicate both PCR primer sequences. The analysed CpG dinucleotides have been numbered relative to the 5\u2019 of the amplicon.Click here for file"} +{"text": "Sus scrofa, Sus barbatus, Sus verrucosus, Sus celebensis, Sus scebifrons, Babyrousa babyrussa, Potamochoerus larvatus, Potamochoerus porcus and Phacochoerus africanus genomes. Specifically, analyses were performed to identify evidence of positive selection using Maximum likelihood (ML) methods within a phylogenetic framework for bacterial and viral sensing Suidae TLR extracellular domains. Our analyses did not reveal evidence of positive selection for TLR3 and TLR7, suggesting strong functional conservation among these two genes for members of the Suidae. Positive selection was inferred for Suidae TLR1, TLR2, TLR6 and TLR8 evolution. ML methods identified amino acid sites of the bacterial sensing TLR1, TLR2, TLR6 and the viral sensing TLR8 to be under persistent positive selection. Some of these sites are in close proximity to functionally relevant sites, further strengthening the case for pathogen mediated selection for these sites. The branch leading to the genus Sus demonstrated evidence of episodic positive selection for TLR1, indicating selection mediated by infectious agents encountered within the specific geographic origin of the Sus. These results indicate that species of the Suidae have positively selected residues within functional domains of TLRs reflective of prior infections. Thus, TLR genes represent candidates for experimental validation to determine their functional role in antibacterial and antiviral activity within members of the Suidae.Members of the family Suidae have diverged over extended evolutionary periods in diverse environments, suggesting that adaptation in response to endemic infectious agents may have occurred. Toll-like receptors (TLRs) comprise a multigene family that acts as the first line of defense against infectious microbes at the host-environment interface. We hypothesized that across the Suidae, positive selection mediated by infectious agents has contributed to the evolution of TLR diversity. Thus, we analyzed Bacterial and viral infectious diseases constitute a significant threat to host survival. Host species have developed various strategies to combat these threats, including the development of innate and acquired immune defenses. The innate immune system provides an immediate defensive response against pathogenic infections while the acquired immune system response to pathogenic infections may require weeks to develop. As pathogens evolve to subvert the host immune response, host immune genes evolve in response. The arms race between hosts and microbial pathogens (host-parasite co-evolution) influences variation in the response to infectious disease agents at individuals, population, species levels and within higher order taxonomic units .TLR1, TLR2, TLR4, TLR5 and TLR6) recognize predominantly bacterial ligands and several fungal and parasite ligands while TLR3, TLR7 and TLR8 are expressed within the endosome and recognize single and double-stranded viral RNA [Adaptive evolution (positive selective) is selective pressure through a change in environment placed on a protein in order to improve the fitness of the organism in that environment . With reiral RNA . TLRs arPrevious studies have documented purifying selection and overSus scrofa (wild boar) is across most of Eurasia while all other species of the genus Sus are restricted to Southeast Asia [Babyrussa babyrussa (babyrussa) is also found in Southeast Asia and Potamochoerus larvatus (bush pig), Potamochoerus porcus (red river hog) and Phacochoerus africanus (common warthog) are restricted to sub-Saharan Africa [Members of the family Suidae have a widespread distribution. The natural occurrence of ast Asia . The Babn Africa . Such diTLR1, TLR2 and TLR6 and viral sensing TLR3, TLR7 and TLR8 as viruses and bacteria are the dominant parasites threatening wild mammals [The aim of this study was to determine whether there is evidence of positive selective pressure in the family Suidae in a phylogenetic framework. We hypothesized that positive selection has contributed to the evolution of bacterial and viral sensing TLRs in the family Suidae. The specific aims of this study were to 1) identify evidence of persistent positive selection at TLRs across members of the family Suidae and 2) to determine whether restricted lineages within the Suidae demonstrate TLR positive selection. We focused on the bacterial sensing mammals . IdentifSus scrofa (wild boar) was represented by a European wild boar (Sus scrofa Europe) and a Asian wild boar (Sus scrofa Asia) to reflect the wide distribution of this species. Southeast Asian suids were represented by Sus verrucosus (javan warty pig), Sus celebensis (sulawesi warty pig), Sus scebifrons (visayan warty pig), Sus barbatus (bearded pig) and Babyrousa babyrussa (babirusa). Suidae species of African origin were represented by Potamochoerus larvatus (bush pig), Potamochoerus porcus (red river hog) and Phacochoerus africanus (common warthog).Ten animals representing 9 species of the family Suidae were utilized in this study. A range map showing the natural distribution of these species is shown in TLR1, TLR2 and TLR6 encoding receptors for bacterial ligands and TLR3, TLR7 and TLR8 recognizing viral ligands were selected for this study. The extracellular domains were the focus since they encode the functional sites involved in pathogen ligand recognition.Sus scrofa reference genome build 10.2 [Sus scrofa reference genome were stored as bam files for each individual animal.DNA extraction, library preparation and sequencing was performed as previously described . Brieflyild 10.2 using thhttp://www.ensemble.org). The accession numbers of sequences obtained from the public databases were TLR1: NM_001031775, TLR2: NM_213761, TLR3: HQ412796, TLR6: NM_213760, TLR7: NM_001097434, TLR8: ENSSSCG00000012118. When a TLR gene was found to have more than one transcript, the longest transcript was chosen. The genomic coordinates of the porcine TLR mRNA sequences within the Sus scrofa genome assembly 10.2 were obtained from Ensemble. Based on these genomic coordinates, sequences of TLR gene orthologs were then retrieved from aligned bam files of Sus scrofa (Sus scrofa Europe and Sus scrofa Asia), Sus verrucosus, Sus celebensis, Sus scebifrons, Sus barbatus Babyrousa babyrussa, Potamochoerus larvatus, Potamochoerus porcus and Phacochoerus africanus to identify TLR gene orthologs. The resulting sequences for each species were then blast screened against the Sus scrofa genome to ensure similarity with the porcine TLR mRNA sequences. Exonic regions were then obtained from these sequences and concatenated to obtain coding sequences. The coding sequences were further trimmed to obtain sequences of the extracellular domain for each TLR in each species. Sequences were aligned using ClustalW 1.81 [Porcine TLR mRNA sequences were obtained from Ensemble database with the number of synonymous substitutions per synonymous site (dS) in a maximum likelihood (ML) framework was used to test for positive selection for every codon, defining a dN/dS ratio (\u03c9)>1 in a codon as evidence of positive selection. First, we determined whether \u03c9 varied among codon sites for each TLR alignment by comparing CODEML program models in PAML version 4 [Comparison of the non-synonymous substitutions per non-synonymous site implemented on the Datamonkey web server were utilized. The Branch-Site REL model estimates proportion of sites under selection along tree branches and allows evolutionary rates to simultaneously vary along phylogenetic branches and sites . The MEMPositively selected sites detected in this study were compared to human TLR Swiss-Prot database to determine their possible link to function. Sites under positive selection were also mapped to three dimensional (3D) protein structures using MuPIT Interactive in orderTLR1, TLR2 and TLR6 and viral sensing TLR3, TLR7 and TLR8 from species within the family Suidae were obtained. The length of the extracellular domains in terms of number of nucleotides of the TLRs ranged from 1668 bases for TLR1 to 2445 bases for TLR7. Amino acid length ranged from 556 amino acids for TLR1 to 792 amino acids for TLR7.The sequences of the extracellular domains of bacterial sensing dN/dS ratio (\u03c9) of some TLR genes varied among codons, implying that selective constraints were heterogeneous between sites. We detected significant (p<0.01 for 2ln\u0394l) heterogeneity of \u03c9 along TLR1, TLR2, TLR6 and TLR8 (2) is relatively smaller than the proportion of sites with evidence of purifying (p0) or neutral (p1) selection. Thus, the majority of sites within the proteins of TLR1, TLR2, TLR6 and TLR8 were functionally constrained. TLR3 and TLR7 sequences did not reveal heterogeneity of selection pressure \u03c9 among their codons and are thus functionally conserved along their entire extracellular domains within the members of Suidae involved in this study.To determine whether selective pressures varied amongst codon sites for each TLR gene, the M0 and M3 models of CODEML program was utilized. Comparison of M0 vs M3 indicated that and TLR8 . For theTLR1, TLR2 and TLR6 and viral sensing TLR8. Specifically, comparisons of nested models available in CODEML program indicated that models including codons with \u03c9>1 (M2a and M8) demonstrated a better fit than did neutral models (M1a and M7) for all the four TLR genes and on TLR2 species branch corresponding to Sus verrucosus (Potamochoerus porcus is under positive selective pressure (TLR1 (codon position 434) in the lineage leading to the genus Sus and the species branch corresponding to Sus verrucosus was identified as under positive selection. Another codon position (codon position 338 in TLR2) was found to be under positive selection in the species branch corresponding to Potamochoerus porcus , 434 (LRR16), 451 (LRR17), 559 (LRR carboxy termini (LRRCT)); TLR2 216 (LRR7), 338 (LRR12); TLR6 183 (LRR6), 334 (LRR12), 452 and 459 (LRR17), 501(LRR19), 554 and 560 (LRRCT); TLR8 178 (LRR5), 388 (LRR13). Positively selected sites which can be inferred to affect protein function based on their location within TLR protein 3D crystallographic structures are shown in TLR1 sites 434 and 451) are within TLR1/TLR2 interface have experienced radical amino acid changes pathogens through heterodimerization with TLR1 and TLR6 [TLR2 also been shown to affect IFN production, making the TLR2 gene evolutionary constrained [Our finding of small proportion of sites of on TLRs ,12. As w on TLRs ,38, more on TLRs , fewer sLR1/TLR2 . Therefoand TLR6 ,41. TLR2strained .Apart from persistent pathogen mediated positive selection acting over long evolutionary time across members of the Suidae, the evolutionary histories of members of the Suidae may have been affected by periodic pathogenic infections confined to specific lineages within certain geographic locations, leading to episodic positive selection within such lineages. Such a signal of adaptive evolution is usually masked by a background signal of purifying selection, which makes their identification difficult. Both Branch Site REL and the MEME methodology implemented in Datamonkey revealed the same lineages were evolving under episodic positive selection, suggesting that sites within these branches are under positive selection. MEME is a recently developed method that allSus clade was found to have undergone episodic positive selection at TLR1 amino acid site 434 indicating that the ancestors of species within the genus Sus had to undergo adaptive changes at this site in response to their environment. With the exception of Sus verrucosus which had methionine at TLR1 site 434, other Sus species had the leucine residue while the African suids and Babyrousa babyrussa had methionine. This finding suggests a possible selective advantage for leucine at TLR1 site 434 in the environment in which ancestors of the Sus species originated. Indeed, methionine seems to be very rare at TLR1 site 434 within the domesticated breeds of Sus scrofa [Sus species stabilizes the TLR1 protein prior to heterodimerization with TLR2 for efficient recognition of diverse bacterial ligands (peptidoglycans and triacyl lipoproteins). This finding of positive selection on branches leading to Sus verrucosus for TLR1 and Potamochoerus porcus for TLR2 requires a cautious interpretation, since only one sequence from one animal is involved in each case. Sus verrucosus is thought to represent a distinct lineage following a deep split with other species of the genus Sus [Sus verrucosus may have exerted selective pressure on its TLR1 gene. Related to Potamochoerus porcus, positive selection on TLR2 gene could partly be due to adaptation to infectious agents within the African rain forest, a location outside of which they are rarely found [Our lineage specific analysis showed that the branch leading to the s scrofa supportienus Sus . It is ply found .TLR1 is within disulphide bonds region. Disulphide bonds are important to the overall function of proteins as they are associated with their folding and stability [TLR1 is adjacent to a glycosylation site. One conclusion would be that positive selection at this site is of consequence as glycosylation of TLRs is thought to influence receptor surface presentation, trafficking and ligand recognition [TLR1, adjacent to a site that leads to impairment of NF-kB activation, suggests a role for this site in regulating inflammatory response to bacterial infection.The case for positive selection within TLR amino acid sites involved in this study is strengthened by the location of specific sites in close proximity to functionally relevant regions. Site 117 of tability . Site 43ognition . The posTLR1/TLR2 heterodimer formation is required for ligand recognition and signal initiation [TLR1 suggest residues at these sites could be under selective pressure to improve the TLR1/TLR2 heterodimer formation\u2026. As was the case with the study of [itiation . Thus chstudy of involvinstudy of . Such siSus are also susceptible to diseases of domestic swine [Results obtained here have implications for present day domestic pigs. African wild suids are susceptible to some viral, bacterial and parasitic diseases of domestic pigs. As European and Asian wild boars are the progenitors of most domestic pigs, it is likely that species of the genus ic swine . Thus, ric swine , are impic swine .TLR1, TL2, TLR6 and viral sensing TLR8 of members of the Suidae that have undergone persistent and episodic positive selection were identified. The evidence of positive selection on the TLR genes reveals that pathogen mediated selective pressure has shaped Suidae TLR evolution. The case for positive selective at amino acid sites is strengthened by location of these sites in close proximity to functionally relevant sites and the radical changes in amino acids at some of these sites across members of the Suidae. Sites under positive selection may have aided in the adaptation of the Suidae to infectious agents that evolved rapidly or that were encountered in new environments.In conclusion, residues within bacterial sensing S1 FigTLR1:Q15399; TLR2:O60603; TLR3: NP_003256.1; TLR6:Q9Y2C9; TLR7:NP_057646; TLR8:NP_619542. Porcine TLR sequences accession numbers: TLR1:NP_001026945; TLR2:NP_998926.1; TLR3:DQ266435.1 TLR6:NP_998925.1; TLR7: DQ647699; TLR8: NP_999352.1. For TLR6, human LRRs were determined from their alignment with murine TLR6. Asterisks, colons and periods under aligned the aligned sequences indicate complete match, strong conservation and weaker conservation of amino acid respectively./ ligand binding residues, d residues involved in dimerization, + residues involved in both ligand binding and dimerization. LRR represents leucine rich repeat. LRRNT represents LRR amino termini. LRRCT represents LRR carboxy termini. Human TLR sequences accession numbers: (DOCX)Click here for additional data file.S1 File(ZIP)Click here for additional data file.S1 TableaExon encoding the extracellular domain.(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file."} +{"text": "Chiari malformation (CM) describes cerebellar tonsillar descent below the level of the foramen magnum. It is commonly associated with syringomyelia and often presents with headache (1). The conventional surgical treatment for symptomatic patients is foramen magnum decompression (FMD) (2) which carries a significant burden of operative morbidity (3). Altered cerebrospinal fluid (CSF) dynamics have been demonstrated in CM patients and CSF diversion has been used as an alternative treatment modality. Patients with chronic headache and radiological evidence of CM represent a therapeutic challenge. In our unit, these are primarily investigated with intracranial pressure (ICP) monitoring aiming to detect objective evaluation of CSF dynamics prior to surgical intervention.In this single centre, retrospective study, CM patients presenting with headaches were extracted from our departmental ICP monitoring database. Patients with an existing CSF diversion shunt or a previous foramen magnum decompression were excluded. ICP monitoring results were analysed with emphasis on median intracranial pressure (mICP) and median pulse amplitude (mPA). Clinical records were reviewed for clinical presentation, surgical management and outcome.16 patients with CM and ICP monitoring were identified. 7 had associated syringomyelia. The mean mICP across the group was 2.97 \u00b1 3.13 mmHg. Mean mPA was 5.23 \u00b1 1.27 mmHg . All patients had mICP < 10 mmHg. 2 patients had mICP < 0 mmHg. 14 out of 16 patients had abnormal pulsatility (mPA > 4 mmHg). 6 patients were treated with primary ventriculoperitoneal shunt (VPS) insertion, 3 underwent FMD, 1 was treated medically with acetazolamide and 5 were managed conservatively. There were no significant surgical complications in either the VPS or the FMD group. At 2 month follow-up, all patients in the VPS group experienced symptomatic improvement. 2 patients in the FMD group experienced symptomatic improvement and one was unchanged.The majority of patients with symptomatic untreated Chiari malformation have increased ICP pulsatility but a \u201cnormal\u201d overall mICP. Raw ICP values are not sensitive in identifying abnormalities of compliance in this patient group. A small group of patients may develop tonsilar descent due to relatively low ICP.VPS insertion may be a safe, effective alternative to FMD for patients with symptomatic CM, even in the absence of hydrocephalus.We propose routine intracranial pressure monitoring in CM patients with headache and, if indicated, cranial CSF diversion as first line management."} +{"text": "Scientific Reports5: Article number: 15678; 10.1038/srep15678published online: 10262015; updated: 01062016There is an error in the Introduction of this Article.Pi54 (formerly known as Pi-kh) was first identified in the Indian rice cultivar HR22 and was cloned from the indica type rice cultivar Tetep.\"\"should read:\"Pi54 was cloned from the indica type rice cultivar Tetep.\""} +{"text": "Low cardiac output syndrome (LCOS) which features classic symptoms of hypotension, tachycardia, oliguria and poor peripheral perfusion after cardiac surgery is not rare in the immediate postoperative period, and usually requires high dose of inotropes and volume replacement. In this setting of critical illness and stress, Critical Illness-Related Corticosteroid Insufficiency (CIRCI) may occur and aggravate hemodynamic instability. However, lack of evidence or guideline exists regarding diagnostic criteria of CIRCI in LCOS or use of steroids.We aimed to investigate clinical features of CIRCI in patients with LCOS after cardiac surgery and assessed the efficacy of steroid use in this subset of patients.We retrospectively reviewed the patients who were suspected as combined CIRCI in the setting of LCOS after cardiac surgery between February 2010 and September 2014. Diagnosis of CIRCI was made by a delta total serum cortisol of < 9\u00b5g/dL after ACTH 250 \u00b5g administration or a random total cortisol of < 10 \u00b5g/dL. Patients who were exposed to steroids before surgery for any medical causes were excluded.28 patients underwent adrenocorticotropic hormone (ACTH) stimulation test in suspicion of CIRCI. Among them, 20 patients met the diagnostic criteria. Mean age was 63.8 \u00b1 12.4 years. Majority of patients (80%) underwent valvular surgeries or pericardiectomy. Patients who were diagnosed with CIRCI (+) showed higher Sequential Organ Failure Assessment (SOFA) score at the time of diagnosis compared to CIRCI (-) patients and higher total bilirubin level . Glasgow Coma Scale (GCS) score was lower in CIRCI (+) group . 3 patients (10.7%) died only in CIRCI (+) group without statistical significance. Among CIRCI (+) patients, only 6 patients (30%) received glucocorticoid therapy at surgeon's discretion. Initial dose of steroid therapy varied from 50 to 240 mg per day. Main reason not to use steroid to the other patients was the concern of infection. Mean blood pressure was elevated by the average of 22.2 \u00b1 8.7mmHg after steroid therapy and duration of inotropic support was significantly shorter in steroid therapy group compared to non-steroid therapy group . Infection developed in 3 patients (15%) only in non-steroid therapy group without significant difference.Critical Illness-Related Corticosteroid Insufficiency should be suspected in patients with Low cardiac output syndrome after cardiac surgery especially when patients shows signs of persistent multi-organ failure such as high SOFA score or total bilirubin level. Glucocorticoid replacement therapy may be considered in patients with Critical Illness-Related Corticosteroid Insufficiency after cardiac surgery to reduce the use of inotropes without increasing additional infection risk."} +{"text": "To study this further, we constructed a bipartite network representing structurally resolved PPIs from all available human complexes in the Protein Data Bank . Analysis of frequently mutated cancer genes within this network revealed that tumor-suppressors, but not oncogenes, are significantly enriched with functional mutations in homo-oligomerization regions . We present two important examples, TP53 and beta-2-microglobulin, for which the patterns of somatic mutations at interfaces provide insights into specifically perturbed biological circuits. In patients with TP53 mutations, patient survival correlated with the specific interactions that were perturbed. Moreover, we investigated mutations at the interface of protein-nucleotide interactions and observed an unexpected number of missense mutations but not silent mutations occurring within DNA and RNA binding sites. Finally, we provide a resource of 3,072 PPI interfaces ranked according to their mutation rates. Analysis of this list highlights 282 novel candidate cancer genes that encode proteins participating in interactions that are perturbed recurrently across tumors. In summary, mutation of specific protein interactions is an important contributor to tumor heterogeneity and may have important implications for clinical outcomes.Recently it has been shown that cancer mutations selectively target protein-protein interactions. We hypothesized that mutations affecting distinct protein interactions involving established cancer genes could contribute to tumor heterogeneity, and that novel mechanistic insights might be gained into tumorigenesis by investigating protein interactions under positive selection in cancer. To identify protein interactions under positive selection in cancer, we mapped over 1.2 million nonsynonymous somatic cancer mutations onto 4,896 experimentally determined protein structures and analyzed their spatial distribution. In total, 20% of mutations on the surface of known cancer genes perturbed protein-protein interactions (PPIs), and this enrichment for PPI interfaces was observed for both tumor suppressors (Odds Ratio 1.28, P-value < 10 Tumor genome sequencing is increasingly being used to inform clinical decisions for cancer patients. This is motivated in large part by the availability of targeted therapies that selectively kill cells harboring specific protein-coding mutations. However, each tumor is characterized by a unique profile of mutated genes with little overlap between patients. Few of these genes can be effectively targeted and nearly one fourth of patients do not harbor any clinically actionable mutations. The recet al.\u22122, ANOVAInteractions were ranked on the basis of the ratio of missense mutations to silent mutations, divided by the ratio of probabilities of the occurrence of a missense versus silent mutation given the codon composition of the interface, similar to the approach in . To deteS1 Figa) Violin plots showing the distribution of the number of PDB structures per protein for oncogenes (n = 56), tumor suppressors (n = 47) and other genes (n = 4600) in our analysis. b) Violin plots showing the distribution of the number of publications in PubMed associated per protein for oncogenes, tumor suppressors and other genes. c) A summary of PDB structures available for cancer genes. \u201cComplex structure\u201d means that there exists at least one structure with a cancer gene bound to another protein or nucleic acid. \u201cProtein structure\u201d means that there is at least a tertiary structure present in the PDB. \u201cDriver genes\u201d simply reflects the number of genes classified as an oncogene or tumor suppressor. d) Violin plots showing the distribution of the number of tumor samples with mutations at a given core residue. e) Violin plots showing the distribution of the number of tumor sample with mutations at a given interface residue. In d) and e), the most frequently altered residues for each set of genes are labeled. f) A density plot showing the distribution of VEST scores for mutations occurring at protein core (green), surface non-interface (blue) and interface residues (pink).(EPS)Click here for additional data file.S2 Figa) surface versus core residues, b) surface interface versus surface non-interface residues.Fisher\u2019s exact tests were performed separately for each set of genes. Shown are the odds ratios and 95% confidence intervals within each set of genes when making comparing the number of mutations located at (EPS)Click here for additional data file.S1 File(XLSX)Click here for additional data file.S2 File(XLSX)Click here for additional data file.S3 File(XLSX)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 TableCancer genes and their interaction partners are listed using Entrez gene ids in columns 2 and 4.(DOCX)Click here for additional data file.S4 Table(DOCX)Click here for additional data file.S5 Table(DOCX)Click here for additional data file.S6 Table(DOCX)Click here for additional data file.S7 Table(DOCX)Click here for additional data file.S8 Table(DOCX)Click here for additional data file.S9 Table(DOCX)Click here for additional data file.S10 Table(DOCX)Click here for additional data file.S11 Table(DOCX)Click here for additional data file.S12 Table(DOCX)Click here for additional data file.S13 Table(DOCX)Click here for additional data file.S14 TableThese tests are performed with residue numbers in binding sites or interfaces.(DOCX)Click here for additional data file.S15 Table(DOCX)Click here for additional data file.S16 Table(DOCX)Click here for additional data file."} +{"text": "Bombus spp.; identified to six colour groups) visiting focal plants of lavender (Lavendula spp.) were carried out by about 13 000 primary school children (7\u201311 years old) from over 400 schools across the UK. 3948 reports were received totaling 26 868 bumblebees.There is an error in the Methodology/Principal Findings subsection of the Abstract. The number of schools is incorrect. The correct sentence should read: Timed counts of bumblebees ("} +{"text": "In addition, detection and optical imaging of 90Y radioactivity using Cerenkov luminescence will also be reviewed. Methods and approaches for qualitative and quantitative 90Y imaging will be briefly discussed. Although challenges remain for 90Y imaging, continued clinical demand for predictive imaging response assessment and target/nontarget dosimetry will drive research and technical innovation to provide greater clinical utility of 90Y as a theranostic agent.This paper overviews Yttrium-90 ( The rare-earth lanthanide, Yttrium-90 (90Y), is almost exclusively a high-energy beta-particle emitting radionuclide used for radiotherapy with a maximum particle energy of 2.28\u2009MeV (average energy of 0.94\u2009MeV) that allows for high dose deposition with an average and maximum soft tissue penetration of 2.5\u2009mm and 11\u2009mm, respectively [90Y has a physical half-life of 64.1\u2009h [90Y-labeled colloid [90Y and its use for radiotherapy are complex and necessitate close collaboration among various medical specialties including nuclear medicine, interventional radiology, medical oncology, and radiation medicine [90Y can be administered via direct injection into a space or cavity , intravenously for peptide receptor radionuclide therapy (PRRT) and radioimmunotherapy (RIT), and intra-arterially for radioembolization (RE) therapy.In general, theranostics are agents that possess diagnostic and therapeutic attributes for personalized patient treatment for various diseases . A commo colloid , 8, tumo colloid , and resmedicine . 90Y can\u03b2\u2212 emitting radioisotopes for osseous metastases [90Y is that it is an almost pure \u03b2\u2212 emitting radioisotope which lacks such gamma photons [90Y, conventional scintigraphic imaging and assessment of the posttherapy distribution of its radioactivity are challenging. This lack of gamma photons led to the development and use of surrogate gamma-emitting radioisotopes labeled peptides and antibodies) with analogous chemical properties as a tracer for 90Y dosimetric assessment and pharmacokinetics [99mTc-) labeled macroaggregated albumin (MAA) is currently used as a surrogate radiotracer for planning 90Y microsphere RE therapy [90Y radiotherapy effects in vivo and such discrepancies may result in unanticipated and unintended toxicities [90Y, subsequent imaging assessment of 90Y radioactivity is an important adjunctive step to assess and verify delivery and dosimetric distribution of the 90Y agent to the target(s) and exclude any nontargeted delivery [90Y radioactivity in both targeted lesions and nontargeted tissues would allow for improved comparisons of radiotherapy outcomes in patients. This review will subsequently discuss the different diagnostic imaging approaches used for therapeutic 90Y radioactivity assessment (Other therapeutic d cancer and Samatastases ) also pr photons . On the kinetics , 17. Lik therapy \u201320. It ixicities \u201323. Givedelivery . Likewissessment .99mTc) have driven the evolution of current planar gamma cameras with optimized collimators and detector crystals for detecting and counting primary photons in discrete energy windows. \u03b2\u2212 particle emission from 90Y produces bremsstrahlung photons which can also be imaged scintigraphically [90Y bremsstrahlung photons are generated when the high-energy \u03b2\u2212 particle is emitted from the 90Y nucleus and then slows while interacting with adjacent atoms. As the electron slows down, its kinetic energy is converted into the continuous energy spectrum of both primary and scattered photons with no dominant energy photopeak for conventional scintigraphic imaging .Conventional scintigraphic imaging and quantification of monoenergetic gamma-emitting medical radioisotopes [90Y-labeled RIT [90Y-labeled microspheres [90Y radioactivity using a phantom model simulating soft tissue extravasation of an intravenous 90Y dose [In 1967, Simon and Feitelberg described posttherapy bremsstrahlung imaging assessment of intra-arterially administered patients . Furthervectomy) , 6, intreled RIT , and intospheres \u201331. In a90Y dose .90Y bremsstrahlung is limited by overlying tissue attenuation, internal photon scattering, variable count rates of emitted bremsstrahlung photons, a wide range of photon energies produced, low spatial resolution (which worsens with increasing source distances to the camera), type of collimation employed , and image processing. In particular, attenuation coefficients may not be constant for the range of photon energies acquired by the gamma camera. Likewise, lower energy bremsstrahlung photons are more likely to scatter than high-energy photons. On the other hand, higher energy photons are more likely to penetrate collimator septae and detector crystals which degrade image quality and limit quantification [90Y bremsstrahlung imaging studies. Subsequent efforts to optimize planar 90Y bremsstrahlung imaging have used Monte Carlo simulation modeling [90Y bremsstrahlung radioactivity is likewise challenging but advances in both qualitative 90Y bremsstrahlung imaging and quantitative 90Y bremsstrahlung imaging have been described using optimized photon energy windows, collimation, attenuation correction, image filtering, and reconstruction [Although technically feasible, image quality for fication , 33\u201337. modeling and thestruction , 37\u201344.90Y radioactivity with limited potential for distinguishing overlapping sources of 90Y radioactivity [90Y bremsstrahlung imaging allows for improved three-dimensional (3D) visualization and anatomic discrimination of discrete adjacent foci of 90Y radioactivity as well as improving the potential for quantification [90Y bremsstrahlung photons but, like planar imaging, SPECT cannot distinguish between primary and scattered bremsstrahlung photons and this limits quantitation [90Y bremsstrahlung SPECT with X-ray computed tomography (CT) allows for attenuation correction and 3D anatomical localization of SPECT findings [It should be noted though that planar quantification is a two-dimensional 2D) assessment of activity . Comparefication . The useD assessmtitation . The fusPECT/CT) . This rePECT/CT) .90Y bremsstrahlung SPECT imaging was described in patients following direct injection of 90Y-colloid and confirmed 90Y bremsstrahlung radioactivity within the complex 3D knee joint space [90Y [90Y-labeled RIT [90Y-labeled microspheres (resin [90Y bremsstrahlung planar and SPECT imaging. The American Association of Physicists in Medicine (AAPM) has issued recommendations for post-RE bremsstrahlung imaging in 2011 which included the use of medium-energy collimation and an energy window of 68\u201392\u2009keV [In 1988, nt space . Subsequace [90Y , intraveeled RIT , 26 and eled RIT , and ints . Table8\u201392\u2009keV .90Y bremsstrahlung imaging can be more readily adopted for whole-body assessment of 90Y distribution [90Y radioactivity [90Y RE assessment of liver radioactivity but image quality is still less than ideal [Given that SPECT imaging requires much more time than planar imaging approaches, planar ribution . On the activity . Recentlan ideal , 65.90Y decays result in therapeutic \u03b2\u2212 particle emission, 32 per million decays result in internal pair production that produces annihilation radiation that can be also imaged in vitro using positron emission tomography (PET) imaging systems [90Y internal pair production represents a promising approach for even more accurate 90Y quantification in vitro and in vivo by minimizing the previously noted challenges associated with 90Y bremsstrahlung imaging [Although the vast majority of systems \u201368. Whil systems . PET det imaging .90Y radioactivity following 90Y-labeled resin microsphere RE for colorectal liver metastases, which demonstrated the feasibility of imaging 90Y in vivo using an existing conventional PET/CT system [90Y radioactivity correlated well with the targeted intrahepatic lesion. Likewise, quantitative assessments of 90Y radioactivity in phantoms could also be performed with further improvement in quantitative accuracy using Time-of-Flight (ToF) PET reconstruction [90Y radioactivity assessment when compared with non-ToF PET imaging systems [90Y bremsstrahlung SPECT/CT imaging [90Y internal pair production PET imaging for patients following direct injection of 90Y [90Y-labeled RIT [90Y-labeled microspheres ..90Y radi90Y internal pair production is limited by its very small branching fraction and therefore necessitates longer acquisition times than traditional positron-emitting radioisotopes which has a branching fraction of 967 per 1000 decays). It was also noted that measureable background radioactivity was dependent upon the PET imaging system used. The presence of a small fraction of radioactive Lutetium-176 (176Lu) within the detection crystals of PET imaging systems contributes to this measureable background radioactivity [176Lu background radioactivity be corrected for in order to obtain any accurate 90Y radioactivity assessment using these PET systems [176Lu background radioactivity is not present on PET imaging systems which utilize bismuth germinate (BGO) detector crystals [90Y radioactivity quantification [90Y radioactivity quantification when compared with LYSO-dependent PET systems due to the slower response rate and poorer contrast performance of BGO PET systems [90Y bremsstrahlung radiation.Image quality for activity , 78. Thicrystals and the fication . It has systems . There a90Y and background radioactivity of some PET imaging systems, PET/CT imaging demonstrates better spatial resolution and image contrast than bremsstrahlung imaging [90Y radioactivity compared with even bremsstrahlung SPECT/CT [90Y internal pair production imaging has been studied in vitro and in vivo using a variety of different PET imaging systems, different acquisition times, and different reconstruction algorithms, no standardized or consensus imaging protocol has been described for 90Y PET/CT imaging studies to date. 90Y internal pair production PET imaging studies. In 2013, Kao et al. [90Y PET/CT imaging following RE therapy in order to (1) confirm successful deposition of the 90Y microspheres within the target lesion(s) and (2) detect any nontarget 90Y radioactivity. In this study, 90Y PET/CT imaging was consistently superior to 90Y bremsstrahlung SPECT/CT imaging in the qualitative assessment of post-RE patients, especially in the detection of nontarget 90Y radioactivity [Despite the low branching fraction for PECT/CT) , 44, 51 SPECT/CT . Althougo et al. describeactivity .90Y is real time detection of Cerenkov radiation (CR), that is, ultraviolet and visible light emitted in the presence of high-energy \u03b2\u2212 particle and positron-emitting radionuclides [\u03b2\u2212 particle or positron. Visible and ultraviolet light photons are emitted as the displaced electrons in the water molecules restore themselves to equilibrium and these light photons can be detected with existing high-sensitivity bioluminescence imaging systems. This optical imaging of CR has been designated as Cerenkov luminescence imaging (CLI) [ in vitro for a number of positron-emitting radioisotopes and \u03b2\u2212 particle emitting radioisotope [90Y is the most efficient medical radioisotope for Cerenkov luminescence production [ in vivo CLI has been performed in mouse models following intravenous administration of 90Y salt solution [90Y-labeled peptide [Another innovative approach for imaging of nuclides \u201383. CR ing (CLI) . Detectand 131I) \u201387. To doduction . In precsolution and 90Y- peptide , 88.90Y radioactivity in vitro and in vivo presents many exciting opportunities. High spatial resolution images of 90Y radioactivity using CLI can be obtained within seconds as opposed to several minutes with conventional planar, SPECT, and PET imaging systems. CLI systems also allow for imaging multiple animals simultaneously as opposed to individually using micro-SPECT/PET imaging systems. These CLI systems are also much less expensive when compared with conventional- or micro-SPECT/PET imaging systems. This CLI approach for the preclinical development of targeted 90Y theranostics will be tremendously enabled for researchers and clinicians. Clinical proof-of-concept has recently been described for radiotherapy using 131I [90Y Cerenkography have been described in the literature.This novel optical imaging approach for noninvasively detecting ing 131I . To date90Y imaging is the lack of consensus guidelines for the technical acquisition, imaging reconstruction, and qualitative/quantitative interpretation of planar, SPECT, and PET imaging by the nuclear medicine community and European Association of Nuclear Medicine (EANM)). An initial consensus guideline would establish the basis for future imaging studies to design, develop, and optimize 90Y imaging approaches and reporting. Likewise, a consensus guideline would describe relevant imaging signs following 90Y radiotherapy for imagers [90Y imaging applications. While some manufacturers have provided assistance and expertise to adapt existing imaging systems for 90Y imaging [90Y image acquisition, image reconstruction, and, ideally, quantification. In addition, new technical advances incorporated into the state-of-the-art PET/CT imaging systems like digital PET/CT and continuous bed motion PET acquisition will need to be methodically assessed for advantages and limitations. Although a single case report on respiratory-gated PET/CT imaging for 90Y RE has been described [90Y PET imaging will also need to be addressed.One current challenge for imagers . Another imaging , most imescribed , the adv90Y imaging has largely focused on 3D modalities like SPECT/CT and PET/CT . Although PET/CT imaging systems are more readily accessible today, 90Y PET imaging may be more challenging to incorporate into routine clinical workflows due to the low branching fraction and corresponding low count rates for 90Y [90Y imaging with PET integrated with magnetic resonance imaging (MRI) [Recently, the trend in d PET/CT . The majstudies) . There ing (MRI) . Given t90Y bremsstrahlung SPECT/CT imaging will continue in the future as (1) a reference standard for comparing different 90Y imaging modalities and (2) a more widely accessible imaging modality for qualitative assessment of 90Y radioactivity. As such, continued technical and methodological advances will likely improve SPECT/CT image quality, consistency, and quantification. Although 90Y bremsstrahlung imaging is better with SPECT/CT than planar imaging, planar imaging approaches may represent a more accessible and less expensive qualitative imaging modality capable of performing faster whole-body assessment of 90Y radioactivity than existing SPECT/CT technology. If any gross irregularity is detected with qualitative planar imaging, the patient could be referred for SPECT/CT or PET/CT assessment. The ever-present limitation of 2D planar bremsstrahlung imaging of 90Y radioactivity is the inability to resolve adjacent foci of 90Y radioactivity in target and nontarget tissues. In terms of patient safety and quality control/assurance during 90Y radiotherapy administration , planar bremsstrahlung imaging may play an important role in the future to document successful administration, confirm systemic circulation for nonembolic agents, and exclude any focal soft tissue extravasation or nontarget 90Y radioactivity. To this end, it has been recently proposed to optimize conventional Anger camera technology for interventional 90Y bremsstrahlung imaging applications [Review of current literature suggests that 90Y assessment is CLI. This technology may help to facilitate rapid and more cost-effective preclinical development of a wide array of targeted 90Y-labeled theranostic agents. One challenge for clinical implementation for CLI is the current requirement for no ambient light within the field of view of the CLI system . Ambient light can saturate the highly sensitive CLI imaging system and obscure the true Cerenkov luminescence emissions. Despite this limitation and challenge, human Cerenkography following 131I radiotherapy has already been described [ in vivo/ex vivo assessment of posttherapy 90Y-labeled target or nontarget lesions using CLI-capable endoscopes and specimen analyzers.Another exciting potential imaging modality for escribed . Future 90Y [90Y-labeled resin microspheres was described [90Y-labeled resin microspheres within the source vial in order to obtain a more homogeneous 90Y activity distribution followed by primary measurement of the triple to double coincidence ratio (TDCR) of the sample using both Cerenkov and liquid scintillation detection techniques. The goals for the MetroMRT project as well as other future collaborations will be to develop and validate new approaches for accurately calibrating, assessing, quantifying, and verifying patient dosimetry related to targeted molecular radiotherapy. Such approaches that are ultimately traceable to a primary standard will enable more accurate individual patient dosimetry.An international collaborative project (metrology for molecular radiotherapy or MetroMRT) has been initiated to address the known variability in absorbed dose for patients following radiotherapy, including 90Y . Recentlescribed . This ap90Y imaging will impact future prospective clinical trials which investigate the efficacy and safety of new 90Y theranostics. The long-term value for improved qualitative and quantitative 90Y imaging will be in confirming targeted delivery of the theranostic agent, evaluating nontarget radioactivity, estimating the absorbed dose to the target lesion(s) and nontarget tissue(s), evaluating and predicting treatment response, assessing the predictive power of existing non-90Y surrogate imaging agents, and promoting personalized medicine.Recognizing and addressing the challenges for multimodality 90Y is a theranostic agent which has been used clinically for direct radiation therapy, RIT, PRRT, and RE but it has been and remains a challenging radiotracer in terms of conventional nuclear medicine imaging approaches. The utilization of 90Y targeted radiotherapies is anticipated to increase. There is continued interest in developing and validating noninvasive imaging strategies to assess both targeted 90Y radioactivity and nontargeted 90Y radioactivity that are readily accessible, easy to implement, easy to interpret, and reported in a concise and consistent manner. In general, the 90Y imaging approaches discussed in this review are compatible with a theranostic paradigm [90Y delivery and provide absorbed dose estimates for the target(s) and nontarget tissues. These novel imaging approaches have the potential to further improve the efficacy of targeted 90Y radiotherapies, provide objective treatment monitoring and assessment, and ensure patient safety. Further innovations in qualitative and quantitative nuclear medicine imaging of 90Y radioactivity will continue to impact posttherapy patient management in this era of personalized medicine. The potential for optical imaging of 90Y radioactivity in vitro and in vivo using Cerenkov luminescence may promote more timely and cost-effective preclinical development of targeted theranostics. Clinical and interventional applications for 90Y CLI are also likely to evolve.paradigm . Intrapr"} +{"text": "The assumption of the presence of diffuse myocardial fibrosis in long standing cyanotic congenital heart disease (CHD) inspired us to noninvasively determine the myocardial extracellular volume (ECV) using contrast CMR (T1 mapping).T1 maps were measured pre and 3-10 minutes after the infusion of 0.15 mmol/kg of gadolinium on 25 subjects. Seven adult patients with longstanding cyanotic CHD and no previous surgical history (aged 16-53 yrs and oxygen saturations of 69-90%), nine normal subjects (aged 14-49 yrs), and nine patients with previously cyanotic CHD after total repair during which a heart lung machine was used (aged 2 months-58 yrs). Images were obtained in a mid-ventricular short axis plane. Late gadolinium enhancement using the phase sensitive inversion recovery (PSIR) sequence was performed to exclude scar areas. The T1 values were measured in two areas of the myocardium, in the septum and in the left ventricular posterior or inferior wall, such that same areas were assessed in every patient in the pre and post contrast T1 scan. ECV was calculated according to (1-hematocrit)*(\u0394R1 myocardium/\u0394R1 blood).Patients with cyanosis had significantly lower ECV percentage than the previously cyanotic patients after total repair . No significant differences were found between patients with cyanosis and normal controls . ECV values were significantly different between the three groups in both septum and LV wall .Long standing cyanosis in congenital heart disease without cardiac surgery does not cause diffuse myocardial fibrosis or expansion of the myocardial extra cellular volume.All authors have no conflict of interest."} +{"text": "The optimal timing and modality of therapeutic intervention during early phases of HIV infection is still debated; in our prospective observational study we evaluated immunological and virological outcome in HIV+ patients treated during acute or recent HIV infection.A total of 25 na\u00efve patients with acute or recent (documented infection within six months) HIV infection were recruited at the Infectious Diseases Units of the University of Milan and Turin from 2009 to 2014. Patients received treatment with two NRTIs+one NNRTI/bPI, with or without an induction phase with an additional fourth drug until HIV-RNA undetectability maintained for six months. Blood samples for HIV-RNA, lymphocyte subsets and tropism assessment were obtained at the beginning of the treatment (BL). Patients underwent subsequent six-monthly follow up for clinical outcome, CD4 cell count and HIV-RNA up to 18 months.2 test). Modification or interruption of therapy for tolerability took place in 4 out of 25 patients, all while receiving four drugs; two patients underwent STI between 12 and 18 months following virological success.Median increase in CD4 cells from 0 to 12 months was greater in patients treated during acute (n=18) versus recent (n=7) infection . This higher value was maintained through 18 months, although failing to reach statistical significance. Patients with acute or recent infection did not significantly differ in virological success (83.3% versus 85.7% at 12 months). We considered CD4 cells gains at six months (multivariate analysis, ANCOVA; Treatment of primary infection appeared to be effective in preserving the pool of CD4 cells in acute more than recent infection. There was no evidence of a different outcome through the addition of a fourth drug to the standard treatment."} +{"text": "Introduction: Current guidelines for the functional evaluation of bioprosthetic heart valves recommend the effective orifice area (EOA) as the product of the trans-valvular stroke volume divided by Doppler derived diastolic time velocity integral (TVI). Phase contrast CMR may offer an alternative imaging modality to assess bioprosthetic valve EOA when Doppler methods are technically limited or unreliable.Our circulatory loop includes a mock ventricle and a heart valve imaging chamber that has been fabricated using MRI-compatible components. In this study 3 different sized stented porcine mitral valve bioprostheses were evaluated replicating three different hemodynamic conditions with forward stroke volume of 70 ml, 90 ml and 110 ml respectively at a beat rate of 70 bpm. Imaging was performed with a 1.5T Siemens Avanto scanner. CMR imaging parameters consisted of 25 phases, slice thickness 4 mm, spatial resolution of 138 \u00d7 256 and temporal resolution of 49 msec. Phase contrast pulse sequences were acquired and peak instantaneous velocities were plotted throughout the diastolic filling period. Stroke volume was measured by MRI-compatible high fidelity flow transducer. The velocity time integral of peak velocity was calculated by tracing the area under the curve and CMR-EOA (cm2) was calculated by dividing the forward flow (cm3) by the MRI-derived TVI (cm). Doppler derived EOA was determined by dividing the stroke volume by the continuous wave Doppler TVI.Bioprosthetic mitral valve diastolic flow area was assessed for 3 different sized valves each at 3 flow volume conditions (N = 9). Doppler-EOA and CMR-EOA were measured and compared for each condition. CMR-derived TVI demonstrated a strong and statistically significant correlation with Doppler-derived TVI . CMR-EOA also revealed a strong and statistically significant correlation with Doppler derived EOA . Mean EOA difference was 0.2 cm2 \u00b1 0.13. The lower temporal resolution of phase contrast CMR velocity determination may have led to the lower TVI values and slightly larger EOA calculation compared to Doppler TVI methodIn this study we report a novel method to determine mitral prosthetic valve effective orifice area using phase contrast CMR values. We demonstrate a strong correlation with the current Doppler standard method to derive EOA. CMR-derived EOA may be an important parameter of prosthetic valve function when Doppler methods are unobtainable or unreliable.AHA beginning grant in aid."} +{"text": "Acute chest pain, ST-changes on EKG and elevation of cardiac troponin in patients without obstructive coronary artery disease represent a clinical challenge. Cardiovascular magnetic resonance (CMR) can be used to diagnose causes other than obstructive coronary artery disease.The aim of this study was to evaluate the usefulness of CMRto diagnoseconditions in the emergency room that otherwise would be consideredas acute coronary syndrome (ACS) in patients with normal coronary arteries.Forty-seven patients with chest pain and/or electrocardiographic changes and elevated troponin concentration occurring in the absence of significant coronary artery stenosis were selected and prospectively submitted to CMR exam in a 1.5T Philips scanner between May 2013 and June 2014. Ventricular function by cine MR with SSFP technique, and myocardial tissue characterization using late gadolinium enhancement (LGE) were evaluated in patients referred to the Emergency room. LGE patterns were analyzed visually by 2 observers and classified as ischemic and non-ischemic .Among 47 patients, all with interpretableCMR exams, diagnosis of acute myocarditis was found in 21 patients (45%), acute myocardial infarction in 9 patients (19%) and Takotsubo cardiomyopathy in 4 patients (9%). Other final diagnoses were hypertrophic cardiomyopathy (7%), coronary embolism (4%), cardiomyopathy (4%), sepsis (2%), aortic stenosis (2%) and noncompaction myocardium (2%). In 34 patients (72%), CMR changed theinitial ACS diagnosis to another final diagnosis. Additionally,2 patients (5%) primarily considered as having myocarditis received a final diagnosis of myocardial infarction.In the study, 77% of patients had theirprimary diagnosis and treatment changed after CMR study. The presence, distribution and pattern of late gadolinium enhancement by CMR were crucial in establishing a precise final diagnosis and appropriately changing patient management.Not applicable."} +{"text": "Drosophila, the transcription factors Pph13 and Orthodenticle (Otd) direct both aspects of differentiation: rhabdomeric opsin transcription and rhabdomere morphogenesis. We demonstrate that the orthologs of both proteins are expressed in the visual systems of the distantly related arthropod species Tribolium castaneum and Daphnia magna and that their functional roles are similar in these species. In particular, we establish that the Pph13 homologs have the ability to bind a subset of Rhodopsin core sequence I sites and that these sites are present in key phototransduction genes of both Tribolium and Daphnia. Furthermore, Pph13 and Otd orthologs are capable of executing deeply conserved functions of photoreceptor differentiation as evidenced by the ability to rescue their respective Drosophila mutant phenotypes. Pph13 homologs are equivalent in their ability to direct both rhabdomere morphogenesis and opsin expression within Drosophila, whereas Otd paralogs demonstrate differential abilities to regulate photoreceptor differentiation. Finally, loss-of-function analyses in Tribolium confirm the conserved requirement of Pph13 and Otd in regulating both rhabdomeric opsin transcription and rhabdomere morphogenesis. Taken together, our data identify components of a regulatory framework for rhabdomeric photoreceptor differentiation in Pancrustaceans, providing a foundation for defining ancestral regulatory modules of rhabdomeric photoreceptor differentiation.A hallmark of visual rhabdomeric photoreceptors is the expression of a rhabdomeric opsin and uniquely associated phototransduction molecules, which are incorporated into a specialized expanded apical membrane, the rhabdomere. Given the extensive utilization of rhabdomeric photoreceptors in the eyes of protostomes, here we address whether a common transcriptional mechanism exists for the differentiation of rhabdomeric photoreceptors. In Drosophila (fruit fly) transcription factors, Pph13 and Orthodenticle, are expressed in photoreceptors of Pancrustaceans, Tribolium (red flour beetle) and Daphnia (water flea), and are capable of executing conserved functions of rhabdomeric photoreceptor differentiation. In particular, Tribolium and Daphnia orthologs are capable of substituting and rescuing the photoreceptor differentiation defects observed in their corresponding Drosophila mutants. Furthermore, loss of function analysis in Tribolium of both Pph13 and orthodenticle genes demonstrate they regulate opsin transcription and morphogenesis of the photoreceptor apical membrane. Our data illuminate a framework for rhabdomeric photoreceptor differentiation and provide the foundation for defining the ancestral regulatory modules for rhabdomeric differentiation and potential modifications that underlie the functional diversity observed in rhabdomeric photoreceptors.Visual systems are populated by one of two fundamental types of photoreceptors, ciliary and rhabdomeric. Each photoreceptor type is defined by the opsin molecule expressed and the final morphological form adapted to house the phototransduction machinery. Here we address whether a common transcriptional mechanisms exists for the differentiation of rhabdomeric photoreceptors. We demonstrate that orthologs of two Drosophila (fruit fly) visual system (reviewed in Rhabdomeric (r) photoreceptors are one of two fundamental types of photoreceptors that have been described Drosophila, two homeodomain proteins have been identified that are critical for regulating r- photoreceptor differentiation. The first, Orthodenticle (Otd), is the Drosophila ortholog of a conserved family of Otd/Otx homeodomain transcription factors, which are essential for head and brain development across species Drosophila eyes, otd is required for both aspects of differentiation Drosophila retinas sensitive r- opsin, and rh5, a blue (B) sensitive r- opsin in the two inner photoreceptors of Drosophila ommatidia rh6, the Drosophila ancestral long-wave (LW) opsin Drosophila ommatidium In Drosophila eye Drosophila, suggesting that the two proteins cooperate and have overlapping functions with respect to photoreceptor morphology rh6 and rh2otd mutants, phototransduction is abolished in Pph13 mutants due to the loss and reduced transcription of several key components of the phototransduction machinery Drosophila Rhodopsin promoters Drosophila Pph13 is necessary and sufficient for driving photoreceptor specific reporter gene expression from the artificial 3XP3 promoter Drosophila, the question we address here is whether Pph13 and Otd functions represent a common regulatory pathway of arthropod r-visual photoreceptor differentiation.The second critical transcription factor is PvuII-PstI homology 13 (Pph13), a paired-class homeobox protein that is similar to the vertebrate Aristaless-related homeodomain (Arx) proteins Tribolium castaneum (red flour beetle), a second insect, and Daphnia (water flea), a crustacean. Together, insects and crustaceans define the superclade Pancrustacea within the Arthropoda Daphnia, Tribolium, and Drosophila r- photoreceptor differentiation would indicate a pathway common to the ancestor that generated both lineages, at least 500 million years ago Tribolium and Daphnia and found only within LW r- opsins. Further, the Tribolium and Daphnia Pph13 homologs have retained similar DNA binding capabilities to their respective endogenous RCSI sites and we confirmed their functional equivalency to direct photoreceptor differentiation in Drosophila photoreceptors by transgenic rescue. The Otd paralogs of Tribolium and Daphnia are comparable in their ability to direct rhabdomere morphogenesis but exhibit differential abilities with respect to r- opsin regulation in Drosophila. Lastly, functional analyses in Tribolium reveal that both Pph13 and Otd homologs are essential for both aspects of photoreceptor differentiation, rhabdomere creation and r-opsin expression. In particular, Pph13 is a critical factor for LW r-opsin expression and Otd2 is necessary for the transcription of UV sensitive r-opsin. In summary, our data identify common components for rhabdomeric photoreceptor differentiation among Pancrustaceans, providing a foundation for defining the ancestral transcriptional mechanisms for rhabdomeric photoreceptor differentiation throughout Bilateria.To examine whether Pph13 and Otd could represent a common set of transcription factors required for r- visual photoreceptor differentiation, we chose to investigate their orthologs from two key nodal species, Drosophila r- visual photoreceptor differentiation was conserved, we investigated the conservation of orthologs in the genome sequences of the distantly related arthropod species, Tribolium castaneum and Daphnia pulexTribolium had been previously shown to possess two paralogs of Otd: Otd1 and Otd2 Parhyale hawaiensisDrosophila were previously considered unresolved due to the low level of sequence conservation outside the homeodomain; within Diptera there has been a reduction to only one otd paralog was incorrect. Further examination of upstream genomic regions revealed that the DNA encoding the 5\u2032 portion of the homeodomain was present. This conclusion was confirmed by RT-PCR (data not shown) and in the genome draft of a second related species: Daphnia magna. The complete Daphnia pulex Pph13 cDNA sequence thus included sequence from previously annotated loci JGI_V11_8835 and JGI_V11_313449. Sequence conservation between Daphnia, Tribolium, and Drosophila homologs was confined to the homeodomain that may be of similar ancient origin based on conservation in non-arthropod invertebrates studies of the development of the axonal photoreceptor connections with lamina neurons predict a model in which the photoreceptors begin to differentiate at the midline and move laterally as they mature Daphnia magna. In Daphnia pulex, there are 27 annotated r-opsin paralogs Drosophila, Daphnia pulex contain representatives of UV, LW and B- light sensitive r-opsins. This diversity includes 23 LW opsins that split between the LOPA and LOPB clades Daphnia magna but for our examination we assayed the expression of a pool of three putative representatives from LOPA Tribolium contains two r- opsins, one of which belongs to the LW opsin subfamily and one of which belongs into the UV sensitive subfamily Tribolium homolog of Drosophila visual G\u03b2 (G\u03b276C), LOC662674 (beetlebase.org), we also detected a RCSI site contained a potential RCSI site Drosophila RCSI sites . These experiments revealed that SI sites . Interesin vivo expression patterns and in vitro binding assays provided strong evidence that Pph13 and Otd regulate r- visual photoreceptor cell differentiation in Drosophila, Tribolium and Daphnia. To test for functional equivalency among the orthologs and, more importantly, the ability to direct photoreceptor differentiation, we examined whether Daphnia and Tribolium orthologs were capable of rescuing the photoreceptor defects observed in Drosophila Pph13 and otd mutants.The Drosophila Pph13 mutants have two distinct characteristics. First, Pph13 is necessary for expression of opsin rh6 against each corresponding mRNA for injection into Tribolium larvae. None of the DsRNAs affected developmental timing, viability, or external morphological structures as compared to mock injections; scanning electron microscopy of the adult eye did not reveal any major effects on the external organization of the compound eyes Daphnia Otd1 is orthologous to Drosophila Otd or Tribolium Otd1, it has the ability to rescue both rh6 repression and rh3 activation whereas Tribolium Otd1 the ortholog of Drosophila Otd rescued only the activation of rh3. Moreover, even though Tribolium Otd1 is expressed in all photoreceptors and has the ability to activate r- opsin transcription in Drosophila, this ortholog is apparently dispensable for r- opsin expression within Tribolium. Interestingly, the three vertebrate Otx homologs also exhibited differential rescuing activities of rh3, rh5 and rh6 regulation otd locus in insect and crustacean lineages contributed to different subfunctionalization trajectories. While the functional diversification that resulted from this appears bewildering, it implies continued evolutionary interchangeability, which may have been key to consolidating all Otd-related functions onto a single homolog during the loss of otd2 in the lineage to dipteran species.Consistent with the evidence of independent duplication events in the insect and crustacean lineages, we find that there is no simple correlation between the expression profiles of Drosophila eye is a good example of this paradigm. Interestingly, our functional analysis in Tribolium reveals differences in how Pph13 and Otd are employed in directing rhabdomere morphogenesis compared to Drosophila. First, within Tribolium, the reduction of either Otd1 or Otd2 generates non-overlapping defects in rhabdomere morphogenesis while the simultaneous knockdown of both genes leads to complete failure of rhabdomere formation. This outcome could result from incomplete subfunctionalization, leaving a limited degree of genetic redundancy in place. Alternatively, the two paralogs may have limited capacities to compensate for the downregulation of the sister paralog via expression level increase. Second, in Drosophila, both Pph13 and Otd are providing independent and overlapping functions to generate the rhabdomere Tribolium, the knockdown of either Pph13 alone or the knockdown of both otd paralogs is sufficient to eliminate rhabdomeres. Thus, there appears to be significantly less redundant control of rhabdomere formation in Tribolium in contrast to Drosophila. Assuming the general conservation of rhabdomere structure target genes, this finding implies evolutionary turnover of Otd and Pph13 dedicated target genes in the context of rhabdomere formation. Given the well defined binding properties of Pph13 and Otd and the conservation of the Pph13 binding sites between Drosophila and Tribolium, it should be feasible to elucidate the diverged distribution of Otd versus Pph13 targets in Tribolium versus Drosophila. Such studies will expand our understanding of the evolution of gene regulatory networks by specifically testing the proposal that downstream network components enjoy greater degrees of evolutionary freedom than intermediate upstream components The activation of structural gene batteries forms the endpoint in the gene regulatory network control of cell differentiation Daphnia Pph13 and otd2 have not been successful. Fortunately, with the advent of TALEN and CRISPR technology Pph13 relates to the diversification of ciliary and rhabdomeric photoreceptors during early metazoan evolution and whether Pph13 or Otd is required for the differentiation of non-visual rhabdomeric photoreceptors, as exemplified by intrinsic photosensitive retinal ganglion cells (ipRGC) Ever since Darwin pondered about the evolution of the eye Tribolium Pph13 and otd2 were constructed from RT-PCR reactions from total RNA isolated from beetle heads. The otd1 cDNA was a gift from Dr. G. Bucher. cDNAs representing Daphnia magna Pph13 and otd2 were constructed from RT-PCR reactions from total RNA isolated from whole adults. The Daphnia magna cDNA of otd1 was isolated by screening an embryonic cDNA library Pph13-GAL4 was generated by inserting the immediate upstream 1.6 kb of genomic DNA extending from the first coding Methionine of the Pph13 locus into pCHS-GAL4. Drosophila strains utilized: cn bw, hzycn bw Pph13uviotdy w; Sp/Cyo; UAS-Flag-otd/TM2uvi; otd-GAL4, Pwiz6/Cyo; TM2/TM6Botdotd rescue experiments females of uvi; otd-GAL4, Pwiz6/Cyo; TM2/TM6Botd were crossed to w; +; UAS \u2013 otd X transgenic lines and only non CyO;TM6B male progeny were examined. For Pph13 rescue experiments w; cn bw Pph13/CyO; Pph13-GAL4/TM6B homozygous flies were crossed to w; cn bw Pph13/CyO; UAS- Pph13 X/TM6B and only non CyO; TM6B progeny were examined.cDNAs representing All cDNAs were cloned into pCDNA 3.1(Invitrogen) and EMSAs were performed as described in pu11, wm26, or v late stage larvae; pu11 and m26 contain a 3XP3-GFP and a 3XP3-RFP reporter, respectively To generate RNAi knockdown animals, 1 ug/ul of total probe, DsRNA was injected into th) of RT reaction and samples were collected at 25 or 30 cycles. All reactions were repeated three times with two independent sets of total RNA. Equal amounts of PCR reactions were analyzed by gel electrophoresis. The list of PCR primers used can be found in Total RNA was isolated using Trizol and DNAase treated. First strand synthesis was accomplished with the Superscript III (Invitrogen) kit. Each reaction contained 2 ug of total RNA and oligo-dT as primers. PCR amplification was performed with 1 ul (1/20Tribolium release 3.0 gene models (http://www.Beetlebase.org/) using TopHat2 version 2.0.9 Tribolium, read counts were normalized across samples using the DESeq package (version 1.12) in R/Bioconductor Tribolium LW and UV opsin are listed in For each condition, duplicate sets of total RNA from entire animals (<12 hours old) were isolated. Stranded RNA sequencing libraries were constructed using the TruSeq Stranded mRNA Sample Preparation Kit according to manufacturer's instructions. Libraries were quantified using the KAPA SYBR FAST Roche LightCycler 480 2X qPCR Master Mix , pooled in equal molar amounts, and sequenced on a HiSeq2000 instrument using a 100 bp paired-end run. HISeq read sequences were cleaned using Trimmomatic version 0.30 Drosophila and Tribolium heads were prepared as previously described Tribolium Pph13 polyclonal antibody, 171, was created by injecting rats with a GST-fusion protein representing amino acids 104\u2013220 of the protein. The Daphnia magna Otd2 polyclonal antibody was prepared in guinea pigs against a bacterially expressed C-terminal portion of the protein (amino acids 235\u2013395). The Daphnia magna Otd1 polyclonal antibody was prepared in rats against a bacterially expressed portion of the protein (amino acids 220\u2013351).The in situ hybridization in Tribolium and Daphnia was performed as previously described Tribolium Pph13 (1\u223620), rat anti- Daphnia Otd1 , guinea pig anti- Daphnia Otd2 mouse anti-Rh3 , mouse anti-Rh5 and rabbit anti-Rh6 . For Otd2, in Daphnia, signals were amplified with the Tyramide Signal Amplification (TSA) system (PerkinElmer). FITC and Rhodamine conjugated secondary antibodies were utilized (Jackson ImmunoResearch). Immunofluoresence studies were performed as previously described Drosophila samples were from newly emerged adults and for each genotype at least three retinas from three different heads were examined.Whole mount RNA Many of the sequences utilized in this study can be found in Figure S1http://homeodb.zoo.ox.ac.uk/).Phylogenetic tree of Orthodenticle and related homeodomain transcription factors. Maximum likelihood tree estimated in MEGA 5.2.2 (TIF)Click here for additional data file.Figure S2http://homeodb.zoo.ox.ac.uk/).zPhylogenetic tree of Pph13/Aristaless and related homeodomain transcription factors. Maximum likelihood tree estimated in MEGA 5.2.2 (TIF)Click here for additional data file.Figure S3in situ hybridizations with sense probes in the developing retina of adult Tribolium. A\u2013C. Dissected retinas 30\u201340 hrs APF of developing Tribolium castaneum adult visual system. None of the sense probes for Pph13 (A), otd1 (B), otd2 (C) demonstrate a specific expression pattern in the developing retina. At this developmental time point it is impossible to discern the developing retina field from surrounding tissue without a molecular marker for the photoreceptors.RNA (TIF)Click here for additional data file.Figure S4Daphnia pulex. Phylogeny of 27 Daphnia pulex opsin proteins inferred using the Neighbor-Joining method with bootstrap test (500 replicates). Scale bar represents genetic distance as number of amino acid substitutions per site. Tree modified from original data presented in Phylogenetic tree of r- opsins of (TIF)Click here for additional data file.Figure S5Daphnia magna. Mixed staged embryos were examined for expression of long wave (LW) A r-opsins , long wave B r-opsins , ultra violet (UV) r-opsin , otd2 , and Pph13 . Circles represent expression in the putative eye. None of the sense probes demonstrate a specific expression pattern in the developing retina.RNA in situ hybridizations with sense and antisense probes in the developing retina of (TIF)Click here for additional data file.Figure S6D. melanogaster, T. castaneum and three D. pulex homologs inferred from Neighbor-Joining method with bootstrap test (500 replicates). Scale bar represents genetic distance as number of amino acid substitutions per site. Loci in red indicate G\u03b2 containing an RCSI site. G\u03b276C-flybase.org, LOC662674-beetlebase.org.Phylogenetic tree of G\u03b2 comparisons. Evolutionary relationships of G\u03b2 proteins of (TIF)Click here for additional data file.Figure S7Daphnia magna Pph13. Electrophoretic mobility shift assays of Daphnia magna Pph13 protein on its endogenous G\u03b2 RCSI site with or without 25 fold excess of cold competitor RCSI sites Click here for additional data file.Figure S8Daphnia magna Otd2 in an otd mutant background. Utilizing the established paradigm of +white tissue\u201d.Expression of (TIF)Click here for additional data file.Figure S9In vivo rescue of Rh6 opsin repression in Drosophila orthodenticle mutant background. A\u2013E. Rh6 opsin protein expression in adult Drosophila retinas. A. Wild-type retina. Rh6 protein accumulates in approximately 70% of the central R8 photoreceptors. B. otd mutant. Rh6 expression is detected in numerous photoreceptors in each ommatidium and accumulation is diffuse due to the disruption of rhabdomere morphology. C\u2013G. Rescue of otd mutant with: C. Drosophila (Dmel) otd. D. Tribolium (Tcas) otd1. E. Tribolium (Tcas) otd2. F. Daphnia magna (Dmag) otd1. G. Daphnia magna otd2. All Otd orthologs, except Tcas Otd1, are capable of repressing rh6 transcription and limiting Rh6 expression in a subset of R8 photoreceptors. Insets represent a magnified view of a region of each panel.(TIF)Click here for additional data file.Figure S10In vivo rescue of Rh5 opsin expression in Drosophila orthodenticle mutation. A\u2013E. Rh5 opsin protein expression in adult Drosophila retinas. A. Wild-type retina. Opsin protein accumulates in the rhabdomeres and thus appears as a tube like structure. B. otd mutant. The absence of Otd activity results in the absence of Rh5 transcription and thus no protein is detected. C\u2013G. Rescue of otd mutant with: C. Drosophila (Dmel) otd. D. Tribolium (Tcas) otd1. E. Tribolium (Tcas) otd2. F. Daphnia magna (Dmag) otd1. G. Daphnia magna otd2. Only with Tcas otd2 did we see the expression of Rh5 in the absence of additional melted signaling (TIF)Click here for additional data file.Figure S11Tribolium otd2 affects rhabdomere biogenesis. Transmission electron microscopy analyses of Tribolium adult rhabdomeres. (A\u2013C) Three additional samples of phenotypes observed with the knockdown of otd2. All samples were from newly emerged adults. Scale bar is 2 um. The removal of otd2 resulted in rhabdomere degeneration as observed by the large separations between microvilli and extension of microvilli into the cell body.Knockdown of (TIF)Click here for additional data file.Table S1Daphnia, Tribolium and Drosophila RCSI sites. Motif search: Motif search for TAATNNNATTA carried out using perl script \u201cmotifsearch.pl\u201d on 1000 bp upstream regions of 27 Daphnia pulex opsin genes and confirmed by eye. Daphnia pulex genome release 1. Due to gaps in genome coverage, we could not determine whether an RCSI site exists in Daphnia r- opsins: LOPB12, LOPB13, LOPB14, and UNOP1.Identification and Sequence of (DOCX)Click here for additional data file.Table S2Quantification of RNAi injections scored for the absence or presence of 3XP3-RFP.(DOCX)Click here for additional data file.Table S3in situ hybridizations.Probes regions for RNAi and RNA (DOCX)Click here for additional data file.Table S4PCR primers for reverse transcriptase-polymerase chain reaction assays.(DOCX)Click here for additional data file.Table S5RNAseq data for Tcas LW and UV opsins.(DOCX)Click here for additional data file.Table S6RNAseq total read counts.(DOCX)Click here for additional data file.Table S7Gene sequences utilized in this study.(DOCX)Click here for additional data file.Movie S1Daphnia magna. The nuclei are counter stained with DAPI.Confocal stack of Otd2 expression (green) in the two visual clusters at 48 hrs after egg deposit (AED) in (MOV)Click here for additional data file."} +{"text": "Streptomyces ambofaciens DSM 40697 was completely determined. The genome consists of an 8.1-Mbp linear chromosome with terminal inverted repeats of 210\u00a0kb. Genomic islands were identified, one of which corresponds to a new putative integrative and conjugative element (ICE) called pSAM3.The sequence of Streptomyces ambofaciens DSM 40697 (http://www.ncbi.nlm.nih.gov/genome/annotation_prok/).We report here the complete genome sequence of SM 40697 , which iS.\u00a0ambofaciens DSM 40697 genome consists of a single linear replicon of 8,137,876\u00a0bp, with long terminal inverted repeats of 212,655\u00a0bp . Such a region showed most of the typical elements of actinomycete integrative and conjugative elements (AICEs [int), excision (xis), replication (repSA), transfer (traS), intramycelial transfer (spdABCD), and regulation (pra). The element presents an average G+C content of 65.25%. It was shown to excise between flanking direct repeats (21 bp) and to circularize when S.\u00a0ambofaciens DSM 40697 was cocultured with strains devoid of pSAM3 .The S.\u00a0ambofaciens DSM 40697 allowed us to map loci involved in early reports of genomic plasticity (reference hasR and hasL [The availability of the genome sequence of and hasL ), were lCP012949. Strain DSM 40697 is available from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ).The genome sequence has been deposited at GenBank under the accession no."} +{"text": "Sepsis lacks a reliable measure of disease activity , 2. TherDetect trends in vital signs and routine biomarker levels during sepsis resuscitation in the emergency department (ED).0) and 3 hours later (T1) and the differences between T0 and T3 (delta) were analyzed.Prospective observational pilot study in the ED of a tertiary care teaching hospital. Adult non-trauma patients with two or more SIRS criteria and suspected infection were included. Blood samples were taken and vital signs measured at admittance to the ED (TIn total data of 99 patients was analyzed. Of these patients, 63 presented with sepsis, 30 with severe sepsis and 6 patients had septic shock. Trends in vital signs and routine biomarker levels are respectively shown in Figure Vital signs and biomarker levels showed descending trends during resuscitation, except for parameters directly affected by treatment modalities. Despite these trends patients clinically improved. Trends in vital signs and routine biomarkers might be helpful in predicting clinical course and response to treatment in sepsis patients."} +{"text": "Patients undergoing THA/TKA were screened pre- & postoperatively with urinalysis (UA) including urine dipstick & microscopy. We hypothesized that: 1) many patients without evidence of urinary tract infection (UTI) receive antibiotics if clinicians base treatment on UA results alone; 2) a protocol for screening patients for UTI would decrease treatment for UTI without increasing surgical site infection (SSI) rates.To identify determinants of treatment for UTI among patients undergoing THA or TKA; to assess the effect of implementing a protocol for screening patients for UTI before THA/TKA on antimicrobial use and SSI rates.We conducted a retrospective cohort study of 200 consecutive patients undergoing THA/TKA from 2/21 - 6/30/2011 & a prospective cohort study of 50 patients undergoing these procedures from 5/21 - 7/17/2012 to identify factors influencing treatment for UTIs. We conducted a before-after study to assess the outcome of implementing a screening protocol.The strongest determinants of pre- or postoperative treatment for UTI were positive leukocyte esterase and urine white blood cell count > 5 . At least 59.7% of patients treated did not have clinical evidence of UTI. The screening protocol was revised such that all patients with a positive LE or nitrite test have urine cultures. Patients were treated for UTIs if the cultures grow > 100,000 CFU of 1 organism. Subsequently, the number of patients receiving antimicrobial treatment for presumed UTI decreased 80.2%; the SSI rate did not increase.A new protocol for diagnosis and treatment of UTIs was associated with a significant decrease in treatment for presumed UTI but the incidence of SSI did not increase. These results suggest that most patients with positive LE do not need treatment for UTI before THA/TKA.None declared."} +{"text": "Bacillus subtilis mutants lacking the SMC-ScpAB complex are severely impaired for chromosome condensation and partitioning, DNA repair, and cells are not viable under standard laboratory conditions. We isolated suppressor mutations that restored the capacity of a smc deletion mutant (\u0394smc) to grow under standard conditions. These suppressor mutations reduced chromosome segregation defects and abrogated hypersensitivity to gyrase inhibitors of \u0394smc. Three suppressor mutations were mapped in genes involved in tRNA aminoacylation and maturation pathways. A transcriptomic survey of isolated suppressor mutations pointed to a potential link between suppression of \u0394smc and induction of the stringent response. This link was confirmed by (p)ppGpp quantification which indicated a constitutive induction of the stringent response in multiple suppressor strains. Furthermore, sublethal concentrations of arginine hydroxamate (RHX), a potent inducer of stringent response, restored growth of \u0394smc under non permissive conditions. We showed that production of (p)ppGpp alone was sufficient to suppress the thermosensitivity exhibited by the \u0394smc mutant. Our findings shed new light on the coordination between chromosome dynamics mediated by SMC-ScpAB and other cellular processes during rapid bacterial growth. Efficient chromosome organization and segregation, as well as maintenance of genome integrity, are essential for accurate transmission of hereditary genetic information. Proteins from the SMC family are key players in chromosome dynamics that involve chromosome condensation and segregation, cohesion of sister chromatids and DNA repair , 2. GenePseudomonas aeruginosa.Whereas eukaryotic genomes encode several functionally specialized SMC complexes , a singlDeinococcus radiodurans and Staphylococcus aureus . To label B. subtilis previously published protocol [-1 32P orthophosphate for 4\u20135 generations. Cells were concentrated by centrifugation (1 minute at full speed), then nucleotides were extracted in 1 M formic acid and loaded on PEI cellulose plates (JT. Baker). Plates were developed in 1.5 M KH2PO4 (pH 3.4), exposed onto a Storage Phosphor Screen, and scanned using a GE Storm Scanner. Spots were quantified using ImageQuant software. All nucleotides were normalized by phosphate number and expressed as molar ratios to GTP.For (p)ppGpp dosage cells were grown in modified phosphate starvation medium [50 mM morpholinopropanesulfonate (adjusted to pH 7.0 with KOH), 0.4 mM KHprotocol was adapS1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file.S3 Fig(PDF)Click here for additional data file.S1 Table(PDF)Click here for additional data file.S2 Table(PDF)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(PDF)Click here for additional data file."} +{"text": "Advances in human genomics have allowed unprecedented productivity in terms of algorithms, software, and literature available for translating raw next-generation sequence data into high-quality information. The challenges of variant identification in organisms with lower quality reference genomes are less well documented. We explored the consequences of commonly recommended preparatory steps and the effects of single and multi sample variant identification methods using four publicly available software applications on whole genome sequence data of 65 key ancestors of Swiss dairy cattle populations. Accuracy of calling next-generation sequence variants was assessed by comparison to the same loci from medium and high-density single nucleotide variant (SNV) arrays.The total number of SNVs identified varied by software and method, with single (multi) sample results ranging from 17.7 to 22.0 (16.9 to 22.0) million variants. Computing time varied considerably between software. Preparatory realignment of insertions and deletions and subsequent base quality score recalibration had only minor effects on the number and quality of SNVs identified by different software, but increased computing time considerably. Average concordance for single (multi) sample results with high-density chip data was 58.3% (87.0%) and average genotype concordance in correctly identified SNVs was 99.2% (99.2%) across software. The average quality of SNVs identified, measured as the ratio of transitions to transversions, was higher using single sample methods than multi sample methods. A consensus approach using results of different software generally provided the highest variant quality in terms of transition/transversion ratio.Our findings serve as a reference for variant identification pipeline development in non-human organisms and help assess the implication of preparatory steps in next-generation sequencing pipelines for organisms with incomplete reference genomes (pipeline code is included). Benchmarking this information should prove particularly useful in processing next-generation sequencing data for use in genome-wide association studies and genomic selection.The online version of this article (doi:10.1186/1471-2164-15-948) contains supplementary material, which is available to authorized users. Taking both polygenic additive (pedigree) effects and genomic (SNV) effects into account, between 71 and 85% of the genetic variance observed in phenotypic traits of interest in cattle can be explained solely by SNV effects. Fueled The translation of raw NGS reads into tangible variants via re-sequencing is a specific, delicate and computationally demanding task and compBos taurus reference genome UMD3.1 is similar in size to the human genome and contains ~2.8 billion base pairs, approximately 10% of which are not assigned to any chromosome).The objective of this study was to investigate which methods and software work best for detection of high quality genetic variants using NGS data in cattle, with a specific focus on single nucleotide variants. Using whole-genome sequence information from 65 individuals, we a) explore the implications of preparatory steps commonly recommended in human analysis, b) compare results of single and multi sample variant detection achieved using four publicly available variant detection software programmes, c) provide a comparison of computational processing time, and d) compare accuracy and completeness of SNVs identified in NGS data by comparing them to genotypes from the same individuals generated with either high- or medium-density SNV arrays, as well as to analyse genome-wide Ti/Tv ratios. Through benchmarking different variant detection methods in cattle, preliminary recommendations for variant identification in other organisms can be extrapolated. Our findings can serve as a reference for choosing variant identification software and can help assess the implication of preparatory steps in NGS pipelines for species with lower-quality or unfinished reference genomes.Approximately 24 billion paired-end reads were obtained for the 65 sequenced animals. An average of 96.8% of these reads (range 86.6% - 98.2%) were mapped to 30 chromosomes of the bovine reference genome assembly UMD3.1. ApproxiThe results of single sample variant detection are in Figure\u00a0InDel realignment had a slight effect on the number of SNVs and InDels identified, with the largest effects observed in UG. InDel realignment reduced the number of SNVs identified , increased the number of InDels detected , and decreased the number of multi-allelic sites identified and UG , but not in SAM are shown in Table\u00a0The total number of SNVs identified by combining single sample results of all 65 animals was higher for PL and SAM than when multi sample variant identification was carried out on all 65 animals simultaneously Table\u00a0. This waSurprisingly, the total number of InDels identified using single sample variant detection methods was also higher for PL (+4%) and SAM (+34%) than when multi sample methods were used. Again, this was not the case for the UG (-10%). In contrast, Liu et al. analysedConsensus SNVs and InDels for multi sample results may provide a simple method to ensure higher variant accuracy, although this approach is computationally intensive. Analysis of variant sets produced in phase one of the human 1000 genomes project showed tBy using default parameters, we did not fine-tune all possible options available in the individual software applications. Nevertheless, using default settings in both single and multi sample variant identification yielded good performance while maintaining output quality. Our goal was to provide an initial overview of methods using the default settings recommended; it should be noted that each dataset must be treated uniquely and alternative parameter settings may deliver more accurate results.If possible, we recommend a consensus approach for variant identification using SNVs identified by all software, which resulted in the highest SNV quality and should be considered the \u201cgolden standard\u201d for variant identification in organisms with lower-quality reference genomes. If computational constraints do not allow a consensus approach to variant identification, the tradeoff between quality and quantity of SNVs must be faced (computation time is discussed in the next section). The UG identified the highest number of SNV in both single and multi sample methods, however the number of false positive SNVs was also highest. SNVs identified with PL had the highest quality of single sample methods, however the number of SNV identified by PL may appear to be low if \u201chidden\u201d SNVs are not split into allelic primitives. SAM identified a good number of SNVs, which were of comparable quality to those identified by PL. Both PL and SAM are likely a good choice of software for organisms with lower-quality reference genomes, as the built-in InDel realignment algorithm seems to efficiently remove false positives, making the use of lower quality resources superfluous.Average computation time required for IR and IR\u2009+\u2009BQSR is depicted in Figure\u00a0InDel realignment and base quality score recalibration had only slight effects on the number of SNVs, InDels and multi-allelic sites identified. The effect of InDel realignment on Ti/Tv ratio was only positive for UG, and the effect of base quality score recalibration on Ti/Tv ratio was negligible (PL) or even slightly negative . Given that computational costs in terms of time are very high, we recommend InDel realignment only in combination with UG. The use of BQSR for organisms with lower-quality resource information seems superfluous until better resources become available.Concordance, measured as non-reference sensitivity (NRS), non-reference discrepancy (NRD), SNV concordance and genotype concordance, was calculated by comparing variants identified in NGS data to array information from the Illumina BovineSNP50 v1 DNA Analysis BeadChip\u00ae (n\u2009=\u200917 samples) and the Illumina BovineHD BeadChip\u00ae using single sample methods was higher than that of multi sample calling for PL and UG and slightly lower for SAM, whereby a consensus approach using results of different software generally provides the highest variant quality. Computation time for single and multi sample methods was similar when calculated on a per-sample basis. These findings can serve as a reference for variant detection pipeline development in various organisms and help assess the value of preparatory steps in NGS pipelines for species with lower-quality reference genomes.m animals were selected with, where p is a vector that contains the percentage of gene pool diversity captured by m animals,m animals and c is a vector representing the average relationship of the m animals selected to the entire genotyped population. The subset of selected sires consisted of key Brown Swiss (n\u2009=\u20097), Braunvieh (n\u2009=\u200917) and Original Braunvieh (n\u2009=\u20098) Simmental (n\u2009=\u200912), Swiss Fleckvieh (n\u2009=\u20094) and (Red) Holstein (n\u2009=\u200917) ancestors that accounted for 74% of the genetic diversity of currently available genotyped populations of these breeds. All animals selected were male. Pairwise identity by descent was estimated for the merged dataset of the 65 sequenced sires by using the \u2013genome function implemented in PLINK on array genotypes[We selected 65 key ancestors of the main Swiss dairy populations with an iterative algorithm, which uses the numerator relationship matrix to rank animals according to percentage of genetic diversity they explain in a given population. Specifienotypes. A heat Sequencing was done at the Helmholtz Center in Munich, Germany in collaboration with the Technical University of Munich. Genomic DNA was extracted from semen samples and sequenced using an Illumina HiSeq2000 . Individual samples were sequenced on individual lanes of the flow cell. The bases of the resulting paired-end reads (101\u00a0bp) were identified with the Illumina BaseCaller; FASTQ files were proSequence alignment was done according to the sequence alignment guidelines for producing binary alignment mapping (BAM) files for the 1000 bull genomes project. BrieflyBoth single and multi sample methods for variant detection were applied. Single sample variant detection was performed with three different software applications: 1) SAM , 2) PL For single sample variant identification, three levels of quality recalibration were compared for each animal and software application: a) no quality recalibration (RAW), b) local realignment around insertions and deletions using the GATK IndelRealigner walker (IR) and c) Computation time required for preparatory steps and variant detection of a 5Mbp portion of BTA24 (single and multi sample methods) was tested on an Intel Xeon E5-2650 machine with two eight-core processors and a total of 256Gb RAM. No parallelism options were used for time calculations. For the whole genome analysis, single sample variant identification was done on a 24-node cluster at Iowa State University. Each computational node has two six-core processors, 64Gb of RAM, and 1\u00a0TB scratch space, for a total of 288 processors and 1536Gb RAM. Chromosomes were split into regions of similar size for parallelization; one chromosome was run per core and optimal threading options in GATK were implemented. Multi sample variant identification was done on the CyEnce cluster of Iowa State University, which includes 288 nodes with 128Gb RAM each.The accuracy and completeness of SNVs identified in NGS data were assessed by comparing them to genotypes of the same animals generated with either the Illumina BovineHD BeadChip\u00ae or the Illumina BovineSNP50 v1 DNA Analysis BeadChip\u00ae . The high-density array contained 774,660 SNVs mapped to the 29 autosomal chromosomes and the X chromosome . For the high-density array, 772,821 SNVs were assigned refSNP cluster ID numbers from dbSNP build 137 using SNFour measures of concordance and discrepancy between array data and NGS data were calculated as described in Table\u00a0http://www.1000bullgenomes.com/. The identified variants were submitted to the Database of Single Nucleotide Polymorphisms (dbSNP) and are available from: http://www.ncbi.nlm.nih.gov/SNP/.All DNA references used were taken from the publically available bovine assembly UMD3.1 available for download from No animal experiments were performed.Alignment and coverage. Total number of lanes, libraries/pool, reads, number of duplicates, number of mapped reads, net number of mapped reads, net number of bases, and net average coverage per animal. (PDF 54 KB)Additional file 1: Variant counts by animal. a) Number of single nucleotide polymorphisms (SNPs) identified per animal using different software and various pre-variant identification processing steps. b) Number of insertions and deletions (INDELs) identified per animal using different software and various pre-variant identification processing steps. c) Number of multiallelic sites identified per animal using different software and various pre-variant identification processing steps. (PDF 77 KB)Additional file 2: Concordance with the Illumina BovineSNP50 v1 DNA Analysis BeadChip\u00ae (n\u2009=\u200917). a) Non-reference sensitivity (NRS) for single nucleotide variants identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineSNP50 v1 DNA Analysis BeadChip\u00ae as a gold standard (BTA1-BTA29). b) Non-reference discrepancy (NRD) for single nucleotide variants identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineSNP50 v1 DNA Analysis BeadChip\u00ae as a gold standard (BTA1-BTA29). c) Single nucleotide variant concordance identified using Platypus Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineSNP50 v1 DNA Analysis BeadChip\u00ae as a gold standard (BTA1\u2012BTA29). d) Single nucleotide variant concordance by genotypes identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineSNP50 v1 DNA Analysis BeadChip\u00ae as a gold standard (BTA1\u2012BTA29). e) Concordance for homozygous reference genotypes identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineSNP50 v1 DNA Analysis BeadChip\u00ae as a gold standard (BTA1\u2012BTA29). f) Concordance for heterozygous genotypes identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineSNP50 v1 DNA Analysis BeadChip\u00ae as a gold standard (BTA1\u2012BTA29). g) Concordance for homozygous alternative genotypes identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineSNP50 v1 DNA Analysis BeadChip\u00ae as a gold standard (BTA1\u2012BTA29). (PDF 82 KB)Additional file 3: Concordance with the Illumina Concordance with the Illumina BovineHD BeadChip\u00ae (n\u2009=\u200948). a) Non-reference sensitivity (NRS) for single nucleotide variants identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineHD BeadChip\u00ae as a gold standard (BTA1-BTA29). b) Non-reference discrepancy (NRD) for single nucleotide variants identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineHD BeadChip\u00ae as a gold standard (BTA1-BTA29). c) Single nucleotide variant concordance identified using Platypus Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineHD BeadChip\u00ae as a gold standard (BTA1-BTA29). d) Single nucleotide variant concordance by genotypes identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineHD BeadChip\u00ae as a gold standard (BTA1-BTA29). e) Concordance for homozygous reference genotypes identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineHD BeadChip\u00ae as a gold standard (BTA1-BTA29). f) Concordance for heterozygous genotypes identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineHD BeadChip\u00ae as a gold standard (BTA1-BTA29). g) Concordance for homozygous alternative genotypes identified using Platypus (Primitives), Samtools, UnifiedGenotyper and Haplotype Caller (single and multi sample variant identification) using variants identified with the Illumina BovineHD BeadChip\u00ae as a gold standard (BTA1-BTA29). (PDF 123 KB)Additional file 4: PipelineCode.(PDF 52 KB)Additional file 5:A schematic overview of variant identification pipelines and methods examined in this study.(PDF 241 KB)Additional file 6:"} +{"text": "Aortic stenosis (AS) is one of the most common valvular heart disease nowadays Independent factors affecting the postoperative outcome had been studied long time ago.Identification of the predictors of left ventricular mass regression after aortic valve replacement is our aim of this study.Randomized selection of 100 patients, underwent aortic valve replacement with a single type of bio-prosthesis (Medtronic Mosaic) for pure aortic stenosis. The study population showed that, 25/100 (25%) patients had prosthesis-patient mismatch of a moderate degree (indexed effective orifice area (IEOA) from 0.65 cm2/m2 - 0.85 cm2/m2). The effect of prosthesis-patient mismatch on the postoperative echocardiographic findings mainly the regression of left ventricular mass after aortic valve replacement and follow up comparison of the unmatched group with the matched group in addition to the other possible related factors through the multivariate analysis was studied.In multivariate analysis, hypertensive patients, preoperative New York Heart Association (NYHA) class >II and a higher preoperative left ventricular mass \u2264250 g/m2 are independent predictors of incomplete left ventricular mass regression. Age and Gender was found to be insignificant predictors. There was a good correlation between the postoperative left ventricular mass regression (LVMR) and the projected indexed effective orifice area. There was a significant reduction of left ventricle (LV) mass in both groups and a significant reduction of LV mass index among Non PPM group while it was of a no significant reduction in PPM.This study shows that in patients with pure aortic stenosis prosthesis-patient mismatch is associated with lesser regression of left ventricular hypertrophy after aortic valve replacement. Hypertension, preoperative (NYHA) class >II and a left ventricular mass \u2264250g/m2 are other independent predictors."} +{"text": "Tert-/- (G5 Tert-/-) mice with shortened telomeres have significant anemia, decreased erythroblasts and reduced hematopoietic stem cell (HSC) populations associated with neutrophilia and increased myelopoiesis. Intracellular multiparameter analysis by mass cytometry showed significantly reduced cell proliferation and increased sensitivity to activation of DNA damage checkpoints in erythroid progenitors and in erythroid-biased CD150hi HSC, but not in myeloid progenitors. Strikingly, Cre-inducible reactivation of telomerase activity restored hematopoietic stem and progenitor cell (HSPC) proliferation, normalized the DNA damage response, and improved red cell production and hemoglobin levels. These data establish a direct link between the loss of TERT activity, telomere shortening and defective erythropoiesis and suggest that novel strategies to restore telomerase function may have an important role in the treatment of the resulting anemia.Telomere shortening is common in bone marrow failure syndromes such as dyskeratosis congenita (DC), aplastic anemia (AA) and myelodysplastic syndromes (MDS). However, improved knowledge of the lineage-specific consequences of telomere erosion and restoration of telomere length in hematopoietic progenitors is required to advance therapeutic approaches. We have employed a reversible murine model of telomerase deficiency to compare the dependence of erythroid and myeloid lineage differentiation on telomerase activity. Fifth generation Telomeres are composed of repetitive TTAGGG sequences located at the ends of chromosomes , 2 and aTelomere erosion is prevented by the activity of the telomerase ribonucleoprotein complex consisting of the reverse transcriptase (TERT), the RNA template (TERC), and the dyskerin (DKC1) protein complex \u20139. TelomTert-/- and Terc-/-, have been useful models for studying consequences of telomere dysfunction on HSC function, proliferative potential, and replicative lifespan during serial transplantation . HS. HS45]. affected . Similaraffected . Flow cyTert-/- mice, although CD34-LKS CD150hi numbers were reduced, the ratio between CD34-LKS CD150hi, lo and negative cells was also altered, with a higher frequency of CD150lo and CD150negative in G5 Tert-/- mice relative to age matched G0 Tert+/- mice cells divides HSCs into myeloid-biased long-term repopulating cells (CD150hi) and myeloid-lymphoid balanced HSCs with low potential for self-renewal (CD150lo) 19, 46], 4619, 4+/- mice . These dTert-/- (TxG5) mice that was absent in vehicle treated mice (Tert-/- mice was not detected (data not shown), this is most likely attributable to the lack of sensitivity of the techniques available. However, it has been established that small increments in telomere length are sufficient to elicit cellular responses [To establish the reversibility of the effects of TERT loss on hematopoiesis, mice were treated with tamoxifen or vehicle control. LSL cassette deletion resulted in a PCR product in tamoxifen treated G5 ted mice . Tamoxifted mice and alsoted mice . Signifited mice . Neutropted mice . Flow cyted mice . These desponses .We used the novel technology of mass cytometry that allows for the simultaneous measurement of 19 surface markers and 13 intracellular markers to furthTert-/- mice in G5 Tert-/- mice as measured by Ki-67 was significantly reduced as was phospho Rb (pRb) expression, while the percentage of non-proliferating Ki-67 negative and pRb negative MEPs increased and phosphorylated \u03b3H2AX (p-\u03b3H2AX). The differences in median levels of expression were restricted to the non-proliferating cell fraction (Ki-67 negative) in MEPs from non-proliferating progenitors using mass cytometry led to the observation that the p-\u03b3H2AX and pATM markers of DNA damage were elevated in proliferating GMPs, whereas they were not elevated in proliferating MEPs. These data suggest, but do not prove, that myeloid progenitors may continue to proliferate in the face of DNA damage, perhaps due to an altered DNA damage checkpoint response. They are also consistent with previous studies demonstrTert-/- mice also showed evidence of a DNA damage response and had reduced proliferation. Thus, the differences in myeloid and erythroid cell numbers may also be partially explained by the reduction in CD150 hi HSC and within CD34-LKS cells in G5 Tert-/- mice. Previous in vitro colony formation studies have shown that the majority of colonies formed from CD150hi CD34-LKS cells were mixed colonies (CFU-GEMM), while very few (<10%) were formed from CD150 negative CD34-LKS cells. The opposite pertained to neutrophil/macrophage colonies that arise predominantly from CD150 negative CD34-LKS cells [Tert-/- BM cells further support this hypothesis. The susceptibility of CD150hi HSC compared to the CD150lo or CD150negative HSC to telomere erosion and consequent DNA damage is somewhat surprising, as CD150 hi cells proliferate at lower rates [Terc-/- mice, promoting lymphoid differentiation [Accumulation of DNA damage also underlies a diminished long term repopulating capability of HSC stem cells with short telomeres , 16. We KS cells . The siger rates . Recentlntiation . It is tTert+/- and G5 Tert-/- mice (data not shown).Recently, telomere dysfunction has been linked with mitochondrial defects as it results in repression of PGC-1 genes and an associated decline in mitochondrial biogenesis in tissues including CD34loLKS cells . AlthougTert-/- mice and the control mice (WT Tert+/+ and G0 Tert+/-) were of mixed background. However, to control for strain-specific effects and to avoid a \u201cfounder\u201d effect, G5 Tert-/- mice as well as the control mice (WT Tert+/+ and G0 Tert+/-) were generated through five generations of breeding of multiple pairs to produce age-matched controls. The absence of erythropoietic defects in any of the control mice strongly suggests that the consistent phenotype in G5 mice is due to lack of TERT activity and shortening of telomeres rather than to any allele variability among the mice.Both the G5 TERT mRNA delivery [In summary, the use of the combined modalities of flow and mass cytometry conclusively demonstrate that telomere length is highly relevant to erythroid maturation. They suggest that proliferating erythroid progenitors are more sensitive to DNA damage checkpoints triggered by telomere shortening, while myeloid progenitors continue to proliferate. It is possible that anemia and myeloid malignancies associated with aging , 57, 58 delivery , for bonS1 File(DOCX)Click here for additional data file.S1 Fig(DOCX)Click here for additional data file.S2 Fig(DOCX)Click here for additional data file.S3 Fig(DOCX)Click here for additional data file.S4 Fig(DOCX)Click here for additional data file.S5 Fig(DOCX)Click here for additional data file.S6 Fig(DOCX)Click here for additional data file.S7 Fig(DOCX)Click here for additional data file.S8 Fig(DOCX)Click here for additional data file.S9 Fig(DOCX)Click here for additional data file.S10 Fig(DOCX)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file."} +{"text": "EGFP (enhanced green fluorescent protein) or nuclear-targeted (n) LacZ) or secreted (human \u03b11-antitrypsin (hA1AT)) reporter genes were intravenously injected into two different mouse strains, followed by analyses of transgene expressions and serum alanine aminotransferase (ALT) levels at different time points. We found that addition of empty particles to the fixed doses of rAAV8 preparations repressed liver transduction up to 64% (serum hA1AT) and 44% (nLacZ) in C57BL/6 mice, respectively. The similar trend in inhibiting EGFP expression together with concurrent elevations of serum ALT levels were observed in the BALB/c mice, indicating that empty particles may also exacerbate side effects of rAAV8 EGFP transduction. Our results suggest that removal of empty particles from rAAV preparations may improve efficacy and safety of AAV in clinical applications.Empty virions are inadvertent by-products of recombinant adeno-associated virus (rAAV) packaging process, resulting in vector lots with mixtures of full and empty virions at variable ratios. Impact of empty virions on the efficiency and side effects of rAAV transduction has not been well characterized. Here, we generated partially and completely empty AAV8 virions, fully packaged rAAV8 lots, and mixtures of empty and fully packaged virions with variable ratios of empty virions. The aforementioned dosing formulations of rAAV8 expressing either cellular ( Recently, combination of AAV8 capsid with self-complementary vector genome to target liver in another hemophilia B gene therapy trial indeed led to sufficient FIX transgene expression and improved bleeding phenotype.14 However, several patients with high doses of AAV8 vector delivery also had transient increases in transaminases associated with increased AAV capsid\u2013specific T cells and decreased circulating hF.IX levels, although such a vector-related immunotoxicity seemed to be resolvable by anti-inflammation steroid regimens of prednisolone.15Adeno-associated virus (AAV), a small single-stranded DNA-containing nonpathogenic human parvovirus, is an efficient gene transfer vehicle for gene transfer to different tissues including liver, without apparent vector-related toxicities.17 Therefore, it has been speculated that nonfunctional empty virions in clinical vector lots may reduce efficiency of therapeutic gene transduction in the liver by competing with the fully packaged therapeutic vector particles for receptor uptake, internalization, and intracellular trafficking; they may also exacerbate vector-related side effects. However, these hypotheses have not yet been formally investigated in in vivo animal studies.Moreover, it is reported that clinical grade AAV vector lots may consist of mixtures of empty and full virions at variable ratios of empty virions (REVs) up to 90%, depending on purification methods.et al.,18 we generated AAV8 capsid preparations with partially empty (PE (60\u201390%)) and completely empty (CE (100%)) virions as well as rAAV8 lots with >99% of fully packaged virions. We mixed the fully packaged rAAV8 vectors expressing cellular or nuclear-targeted (n) LacZ) and secreted (human \u03b11-antitrypsin (hA1AT)) reporter genes, respectively, with either CE or PE virions at different REVs to simulate different scenarios of REVs with real rAAV8 clinical lots. We then intravenously dosed adult male mice of two different strains with the vector preparations. At different time points after injections, we analyzed the serum hA1AT levels , EGFP (BALB/c), and nLacZ (C57BL/6) expression in the liver sections in the corresponding groups as well as serum alanine aminotransferase (ALT) levels in all groups of the treated mice. Our results revealed that as the percentages of empty virions increase within a certain range in the dosing vectors: (i) transgene expression decreases (as much as 70%) for all three reporter genes in the both strains of mice and (ii) serum ALT levels elevate (as much as threefold) in BALB/c mice treated with EGFP vector. Also, the empty capsids generated in the vector production/purification process appear to be more toxic than those produced in the absence of vector genome plasmid. In summary, our study substantiated the negative impact of the empty virions in dosing AAV vectors on the gene transfer efficiency and total viral particle dose-limiting side effects and highlighted the importance of removal empty particles from clinical grade rAAVs to further improve the efficacy and safety of rAAV gene therapy.Here, using the method described by Ayuso 19 in CE (hA1AT genomes and EGFP genomes) AAV8 capsids well as fully packaged rAAV8nLacZ of each sample were analyzed by sliver-stained sodium dodecyl sulfate\u2013polyacrylamide gel electrophoresis, which showed that viral protein (VP)1, VP2, and VP3 capsid proteins were the only detectable protein components in all preparations to examine morphology of negative-stained virions19 in CE and PE of highly purified rAAV8nLacZ vector consisting of >99% of fully packaged viral particles with none, and 44% reductions in nLacZ transduction as compared with fully packaged rAAV8-nLacZ alone dose and a fixed REV at 75%. This experiment generated the following findings. First, at all GC doses tested, only the lowest dose (3\u2009\u00d7\u2009109) seemed to show significant repression of hA1AT transduction by AAV8 CE particles at all six time points tested inhibition of hA1AT expression by AAV8 CE particles became statically significant, 33\u201334% reductions of hA1AT expression were also noticeable in the 1\u2009\u00d7\u20091010 and 3\u2009\u00d7\u20091010 GC dose groups .As aforementioned, one of the possible mechanisms for the interference of rAAV23 First, although ALT elevations in the group treated with fully packaged rAAV8EGFP only were not as dramatic as what were previously reported,20\u201322 which may be attributed to possible differences in purification methods, serum ALT elevations were indeed observed as early as 1 week after vector injection, peaked at day 14 and not declined to those in the phosphate-buffered saline control group until the third week and (ii) at a REV of 75%, exacerbation of vector-induced transaminitis was seen with empty AAV8 particles derived from rAAV8 production processes but not from the process without vector plasmid were much more pronounced than those with PE AAV8 particles that contained much less rAAVEGFP genome-bearing virions (<10%), even though EGFP is much more immunogenic and toxic than hA1AT were more immunotoxic than the PE capsids containing less than 10% of rAAV8EGFP genome-bearing virions, even though EGFP is known to be much more immunogenic than hA1AT in BLAB/c mice excessive empty capsids are immunotoxic in rAAV liver gene transfer and (ii) PE capsids as a by-product of rAAV production, which contain AAV virions packaged with full or partial viral genomes, are more immunotoxic.The main objective of our current study was to substantiate the negative impact of empty virions on liver-directed gene transduction by rAAV8 but not to elucidate underlining biological mechanisms which is largely unknown at this point. However, as both PE capsids isolated from rAAV8hA1AT and rAAV8EGFP production processes exacerbate transaminitis elicited by rAAV8EGFP liver transduction in BALB/c mice, it is unlikely that the side effects of PE capsids are transgene derived. Among other possibilities, it is plausible that capsid-specific cytotoxic CD8B/c mice . Using t25 This hypothesis was further supported by some new insight for structural biology of AAV capsids. In fact, encapsidation of full or partial AAV genomes may lead to capsid conformational changes, the unique N terminus of the capsid VP1 (VP1u) which contains a phospholipase A2 domain, and two nuclear localization signals could be efficiently externalized through fivefold pore.26 Such conformational change on capsid structure exposes both phospholipase A2 domain and nuclear localization signal domains and facilitates endosome escape and viral infection.25 In contrast, the structural conformation of the CE AAV capsids prevents externalization of unique N terminus of the capsid VP1 in the late endosomes and, therefore, endosome escaping and nucleus trafficking, subsequently resulting in poor antigen presentation. Additionally, inefficient uncoating of CE AAV virions as compared with PE particles could be another possible reason for their lower antigen presentation and immunotoxicity.26 Nonetheless, further studies may be warranted to elucidate those potential mechanisms.A possible mechanism for the differences between CE and PE viral particles in side effects could be related to structural biology and intracellular trafficking and processing of AAV capsids. It was speculated that CE AAV capsid particles are inefficient in endosome escape and therefore less efficient in capsid antigen cross-presentation.17 highlighting its potential clinical benefits in human gene therapy studies. The another novel design of this strategy was to abolish the receptor-binding capability and reserve the neutralizing ability of the empty capsids by site-directed mutagenesis, specifically titrating out host capsid response to rAAV without competing with rAAVs for liver transduction.17 Nevertheless, as indicated in the report, this strategy will still face the challenge of crossly presented capsid antigens from overloaded empty capsids, suffering the consequences of adaptive immunotoxicity.17 However, the findings from our study might have shed some lights on designing this AAV empty decoy strategy, suggesting that the use of CE instead of PE capsids as decoys will not only overcome host immunity but also potentially reduce cross-presentation of excessive capsids and potential CD8+ T-cell response. Therefore, it may be safer and beneficial to add noninfectious mutant CE capsid decoys instead of the PE particles to the therapeutic dosing rAAV vector to blunt the preexisting neutralizing antibodies and reduce potential side effects of capsid decoys in gene therapy recipients. This approach is simple and safe and can be potentially used for vectors derived from all AAV serotypes, which should greatly expand the utilization of AAV vectors in the patients with positive neutralizing antibodies.On the other hand, empty capsids in rAAV preparations may play a beneficial role in overcoming preexisting B-cell immunity to rAAV capsid and enabling rAAV transduction in the patient population that are not acceptable to rAAV gene therapy. This highly innovative concept was clearly demonstrated in a recently report where empty capsids were deliberately added as capsid decoys to the rAAV8 dosing formulation to overcome preexisting humoral immunity against systemically delivered rAAV8,hA1AT dosing formulations at a REV of 75% to study rAAV8 transduction in C57BL/6 mice. Our preliminary data showed no significant inhibition of rAAV transduction by CE AAV2 virions (data not shown). Another issue that remains unclear with our study is the narrow dose range at which statistically significant inhibition of rAAV8hA1AT transduction can be detected in C57BL/6 mice as well as nuclear-targeted LacZ (nLacZ) and EGFP report genes have been described by Ayuso et al.18 The transgenes were driven by chicken \u03b2-action promoter in these constructs. An AAV8 preparation with 100% complete empty (CE) virions was produced in the same way except for the exclusion of vector genome plasmid (cis plasmid) in the transfection step. PE AAV8 virions were produced from the empty virion fractions (low density of ~1.3\u2009g/ml) collected after two steps of CsCl gradient centrifugation in the purification processes for rAAV8EGFP and hA1AT vectors (high density of ~1.4\u2009g/ml). The REV of viral particles was characterized and semiquantified by high-resolution transmission EM and silver-stained sodium dodecyl sulfate\u2013polyacrylamide gel electrophoresis as previously described.19 All the AAV8 vectors and empty virions were produced by the Viral Vector Core at University of Massachusetts Medical School Gene Therapy Center .Preparation and purification of empty virion-free single-stranded rAAV8 vectors expressing secreted human \u03b11-antitrypsin were purchased from Harlan Laboratories . Different vector dosing formulations were administered into C57BL/6 or BALB/c mice intravascularly via tail vein injections. Mouse sera were collected at different time points for detection of A1AT expression and ALT levels. The mouse liver was collected at different time points for detecting GFP and nLacZ expression.31 Rabbit anti-human A1AT antibody and human A1AT standard were purchased from Sigma-Aldrich . Horseradish peroxidase\u2013conjugated goat anit-A1AT antibody was obtained from Fitzgerald . Other reagents for enzyme-linked immunosorbent assay were purchased from KPL . The samples were measured for optical density (OD) at 405 and 600\u2009nm in a luminometer . The \u0394OD (OD405\u2013OD600) was used for data analyses, which are based on a linear regression comparison and interpolation of unknown samples against standard dilutions.An enzyme-linked immunosorbent assay\u2013based assay was used to detect serum hA1AT level as described previously.31 the EGFP transduction efficiency was measured directly by GFP imaging in fixed sections using a Leica DM 5500B fluorescence microscope made by Leica microsystems . LacZ reporter gene expression on X-gal histochemically stained nonfixed liver sections was examined under a light microscope. Images from five visual fields were analyzed quantitatively using ImageJ analysis software . Transgene expression was assessed as total area of green fluorescence (GFP) or blue staining (LacZ) (pixel)2 per visual field (mean \u00b1 SE).Fresh or fixed optimal cutting temperature\u2013embedded liver sections from three hepatic lobes collected from the mock- and vector-injected mice at different postinjection time points were mounted on slides. As described in previous publication,The ALT levels in mouse sera were detected by ALT detection kits purchased from Teco Diagnostics . This colorimetric method for ALT detection is based on dinitrophenylhydrazine formation and is a modification of the classical Reitman Frankel colorimetric end point reaction. The samples were analyzed for OD at 505\u2009nm in a luminometer (BioTek Instruments).t-test or analysis of variance was used to compare between test results and the control, and they were determined to be statistically significant.Student\u2019s"} +{"text": "We sought to determine in a swine model whether a novel echocardiography method, speckle tracking echocardiography (STE), could determine deterioration in right ventricle (RV) function induced by escalating levels of positive end-expiratory pressure (PEEP), and to compare STE with RV fractional area change (FAC). Acute cor pulmonale and hypotension can be induced by high levels of PEEP used in the management of acute respiratory distress syndrome . Quantif2O). RV images were obtained using intracardiac echocardiography and were analyzed offline for FAC and RVfwS .Ten pigs, 40 to 90 kg, anaesthetized, fully mechanically ventilated at 6 to 8 mg/kg were subject to stepwise escalating levels of PEEP at 2-minute intervals , FAC and RVfwS . Paired t tests indicated significant reductions in RVfwS with each step increase in PEEP. FAC only showed significant deterioration at 15 cmH2O PEEP. Significant hypotension (a decrease of more than 20 mmHg) occurred after 10 cmH2O PEEP. RVfwS decreased by a larger extent and earlier than FAC and mean blood pressure with increasing levels of PEEP.Escalating levels of PEEP were strongly associated with significant reductions in mean blood pressure (RVfwS measured by STE is a sensitive method for determining RV dysfunction induced by PEEP. RVfwS displays a stronger association, greater deterioration and earlier reduction than FAC and mean blood pressure with escalating levels of PEEP. This potentially has interesting implications for the role of STE in managing PEEP levels in critically ill patients with acute lung injury."} +{"text": "Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV). Transient expression studies on ten amiRNAs targeting 21\u00a0nt conserved sequences of P1(CBSV and UCBSV), P3(CBSV and UCBSV), CI(UCBSV), NIb(CBSV and UCBSV), CP(UCBSV) and the un-translated region (3\u2032-UTR) were tested in Nicotiana benthamiana. Four out of the ten amiRNAs expressed the corresponding amiRNA at high levels. Transgenic N. benthamiana plants were developed for the four amiRNAs targeting the P1 and NIb genes of CBSV and the P1 and CP genes of UCBSV and shown to accumulate miRNA products. Transgenic plants challenged with CBSV and UCBSV isolates showed resistance levels that ranged between \u223c20\u201360% against CBSV and UCBSV and correlated with expression levels of the transgenically derived miRNAs. MicroRNAs targeting P1 and NIb of CBSV showed protection against CBSV and UCBSV, while amiRNAs targeting the P1 and CP of UCBSV showed protection against UCBSV but were less efficient against CBSV. These results indicate a potential application of amiRNAs for engineering resistance to CBSD-causing viruses in cassava.Artificial miRNAs (amiRNA) were generated targeting conserved sequences within the genomes of the two causal agents of Cassava brown streak disease (CBSD): RNA silencing is a gene regulatory mechanism that controls transcript levels, either by suppressing transcription of messenger RNAs , or by activating a sequence-specific RNA degradation process . RNA silUgandan cassava brown streak virus (UCBSV) and Cassava brown streak virus (CBSV); family Potyviridae, genus Ipomovirus, and poses a major threat to cassava production in East and Central Africa (Bemisia tabaci (Gennadius) and dissnnadius) . Transgennadius) . The impN. benthamiana. Virus sequences for targeting by amiRNAs were identified using Clustal W (Lasergene Version 8) by aligning NCBI accessions of CBSV and UCBSV, namely GQ329864, GU563327, FN434437, FN434436, GQ169761, GQ169760, GQ169759, GQ169758, FJ185044, FJ039520, FN434109 and HM181930. Conserved oligo sequences of \u223c21\u00a0nt in size were identified from the P1, P3, CI, NIb and CP genes and 3\u2032-UTR of CBSV and UCBSV (Supplementary Table 1). Sequences with a 5\u2032-A were preferentially selected to match the sequence requirements of amiRNAs, as previously described by A. thaliana, using primers shown in Supplementary Table 1. PCR-amplified DNA fragments of the amiRNA precursor were subsequently ligated into shuttle vector CGT11003 (Cassava vein mosaic virus (CsVMV) promoter and P1 and CP genes (UCBSV) in a manner similar to that reported previously (P1(CBSV) and NIb(CBSV) showed 25\u201365% resistance against CBSV across the seven T1 transgenic lines tested, with the four lines targeting P1(CBSV) and the three lines targeting NIb(CBSV) showing \u226550% protection against this CBSD causal agent (P1(CBSV) and NIb(CBSV) were also resistant against challenge with UCBSV at levels comparable to those against CBSV showing \u226550% resistance to this pathogen (P1(CBSV) had more amiRNA expressing lines conferring resistance against CBSV and UCBSV, with three out of four lines tested showing \u226550% resistance against both pathogens. None of the other constructs tested had lines expressing amiRNAs with resistance \u226550% against both pathogens.amiRNA constructs targeting (UCBSV) , which wthamiana in additthamiana , was inc to soil with vireviously . amiRNA al agent A and C. nst CBSV A and C. nst CBSV . Three ppathogen B and D. Cucumber mosaic virus (CMV) and Potato virus Y (PVY) and Potato virus X (PVX) amiRNA-mediated resistance the level of expression of some amiRNAs was below that detectable by Northern blot analysis; and (2) a mechanism possibly mediated by non-cleavage of the target is operational. While cleavage is the most predominant mechanism of action in plants , recent CP is the most conserved, while Ham1 is the least conserved between CBSV and UCBSV isolates examined to date were tolerant to the virus construct including 1 or 2 mismatches showed different possible combinations of siRNAs against CBSV and UCBSV (Levels of disease resistance obtained by expressing amiRNAs in this study were generally 50% lower than those previously reported through expression of the \u0394FL-UCBSV CP hairpin siRNA construct . This wand UCBSV . Since ind UCBSV , durablePI and Nib for co-expression within the same plant and investigating the efficacy of this approach within transgenically modified cassava.In conclusion, we report that expression of amiRNA targeting conserved genes in the CBSV and UCBSV genomes results in resistance against these viruses. These results add another potential source of resistance against CBSD-causing viruses in cassava. Future studies will focus on stacking amiRNA expression cassettes such as The authors have no conflict of interest to declare."} +{"text": "Cigarette smoking can harm fertility, but the existing research has targeted primarily on ovarian follicles, embryos or sex hormone. In this study, we tested cigarette smoke extract on ovulation, oocyte morphology and ovarian gene expression associated with inhibition of oxidative stress using C57BL/6 mice. Mice in the experimental group were administered a cigarette smoke extract (CSE) solution (2 mg/ml) orally daily, while the blank control group was given dimethylsulfoxide (DMSO). A positive control group (menadione) was used that received an intraperitoneal injection of 15 mg/kg menadione in oil solution daily. We found that the CSE group manifested a reduced diameter of zona pellucida-free oocyte (ZP-free OD) and a morphologically misshapen first polar body (PB). Our results suggest that CSE exposure is associated with a shrink size and poor quality of oocytes. Quitting smoking is a wise choice to ensure good fertility. B[a]P, a component of cigarette smoke, caused few of ovarian follicles 2] and hydrogen peroxide [H2O2], is a physiological process and occurs in the cell mainly during the mitochondrial energy metabolism. O2 is transformed into a more stable ROS, H2O22O2 concentrations in the cytoplasm reach above the physiological threshold, it can be removed by cytosolic antioxidant systems of the cell. These antioxidant defense mechanisms may include both enzymatic such as catalase, glutathione peroxidase (GPx) Besides, production of reactive oxygen species (ROS), which include superoxide anion p exposure could induce oxidative stress in ovarian follicles. Similarly, Gannon Tobacco smoke as a source of exogenous pro-oxidants, such as ROS and free radical generators, can cause oxidative damage In vitro, B[a]P was shown to inhibit gap junction formation The effects of cigarette smoke on oxidative stress are well known as are the effects of smoke on cellular apoptosis 2) level during ovarian stimulation in IVF 2 synthesis were demonstrated in-vitro maturation, IVF, or in-vitro culture The studies about the effect of smoke on oocyte are few. Smokers present a lower estradiol in each group to prove this standpoint in the future.Synthesizing all the researches mentioned above, it's reasonable to suppose that cigarette smoking may potentially emerge a lower rate of fertility and successful pregnancy, producing oocyte of poor quality, through oxidative stress. For further study, we will perform In the research from Whitcomb Combined with many evidences that smoke and its component causing follicle depletion According to our study, we suggested that CSE exposure was associated with a shrink size and poor morphology of oocytes and oxidative stress maybe the underlying mechanism. We certainly recommend that quitting smoking is a wise choice to ensure good fertility.Dataset S1Data of ovulation quantity.(SAV)Click here for additional data file.Dataset S2Data of ZP thickness.(SAV)Click here for additional data file.Dataset S3Data of ZP-free OD.(SAV)Click here for additional data file.Dataset S4Data of PVS.(SAV)Click here for additional data file.Dataset S5Data of OD.(SAV)Click here for additional data file.Dataset S6Data of PB.(XLS)Click here for additional data file.Dataset S7Ct of 3 replication of ACTIN.(SAV)Click here for additional data file.Dataset S8Ct of 3 replication of GSTM1.(SAV)Click here for additional data file.Dataset S9Ct of 3 replication of GSTM2.(SAV)Click here for additional data file.Dataset S10Ct of 3 replication of GSTP1.(SAV)Click here for additional data file.Dataset S11Ct of 3 replication of HMOX1.(SAV)Click here for additional data file.Dataset S12Ct of 3 replication of NRF2.(SAV)Click here for additional data file.Dataset S13Ct of 3 replication of SOD2.(SAV)Click here for additional data file.Dataset S14Ct of 3 replication of GSTA3.(SAV)Click here for additional data file.Dataset S15Ct of 3 replication of GCLC.(SAV)Click here for additional data file.Dataset S16Ct of 3 replication of GCLM.(SAV)Click here for additional data file."} +{"text": "Wdr13 knockout mice, we show that this gene is important for proper placental development. Wdr13, a X-linked gene, expresses in multiple trophoblast cell types of placenta and the mutant placenta had reduced size after 17.5 dpc due to reduction of junctional zone (JZ) and labyrinth zone (LZ). We observed reduction in levels of angiopoietin-2 and cd44 mRNA in Wdr13 mutant placenta as compared to that in the wild type. Our findings show that Wdr13 is required for normal placental development and cell differentiation. Wdr13 heterozygous female placenta when the mutant allele was of maternal origin showed similar defects as those in case of Wdr13 null placenta. Using two types of heterozygous females carrying either maternally and paternally derived mutant Wdr13 allele we provide genetic evidence that development of placenta determines body weight of mice for the entire life.Placental development is essential for implantation and growth of foetus in the uterus of eutherian mammals. Numerous growth factors are responsible for placental development and cell lineage differentiation. Gene knockout mice have shown role of various genes in the placenta. Here using Trophoectoderm surrounding the blastocyst, differentiate into variety of trophoblast cell types. The trophoectoderm in direct contact with inner cell mass (ICM) is known as polar trophoectoderm and other which is not in contact with ICM is known as mural trophoectoderm124Placenta is an essential organ for normal embryonic development in uterus. In the mouse uterus, soon after implantation at around 4.5 Tpbpa positive and negative EPC progenitors. However spiral artery-associated TGCs (SpA-TGCs) are originated from Tpbpa positive and sinusoids TGCs (S-TGCs) from Tpbpa negative EPC progenitorsTpbpa is a marker for both SpT and GCs. The origin of GCs is not understood properly. Some studies suggest that GCs originate from SpT while others suggest that GCs may originate directly from progenitors present in EPC8All the trophoblast giant cells (TGCs) are polyploid in nature with endocrine activity. However, they differ in their polyploidy, developmental origin, gene expression profiling and locationWdr13 is a member of WD-repeat protein gene family and it expresses in various tissues in mouse and humans1010Wdr13 mutant mice have slightly lower body weight at one month of age and become mildly obese with age as compared to Wild type littermates12Wdr13 is expressed in placenta and has a role in placental development. Further, we took advantage of the preferential inactivation of paternally derived X-chromosome in the female placentaWdr13 heterozygous placenta- when the mutant allele is of maternal origin, have similar defects as in the case of Wdr13 null placenta and provide genetic evidence that placental development affects the lifetime body weight.Wdr13 genotype on the weight of embryos. To generate Wdr13 null and wild type female and male mice, we crossed Wdr13 heterozygous female with either hemizygous male (XWdr13Y) or wild type male (XY) [XWdr13designates an X chromosome carrying Wdr13 mutation and maternally derived X chromosome is designated first]. At 19.5 dpc the weight of Wdr13 null mice were significantly lower than wild type controls .dpc Wdr13 null mice as compared to wild type mice may either be due to deficiency of Wdr13 gene felt in placenta or due to deficiency of this gene in embryo or both. To distinguish among these possibilities, weight of Wdr13 null and their wild type littermate\u2019s embryo/placenta were measured at various time points. At 15.5 dpc there was no difference either in placenta or in the embryo weights between these two genotypes placenta/ embryos and hemizygous males were crossed with wild type females to generate paternally derived heterozygous (XXWdr13) placenta/embryos. These crosses provided us two types of heterozygous female embryos: those developing in Wdr13 null placenta due to inactivation of wild type allele on paternal X chromosome and the maternal allele being the mutant, and those developing in heterozygous placenta where mutant allele was present on inactivated paternal chromosome and wild type allele on maternal chromosome was intact. Indeed, qRT-PCR and western blot analysis of placenta and embryo confirmed this duality were transferred to wild type CD1 pseudo pregnant female. At 17.5 dpc pregnant mice were dissected and weight of placenta/embryo was measured. Consistent with our above-mentioned results embryo transferred Wdr13 null mice placenta showed less weight as compared to wild type littermates staining was performed at 17.5 dpc. MT staining showed significant reduction in the number of S-TGCs in Wdr13 mutant placenta as compared to that in wild type placenta (Wdr13 mutant as compared to wild type placenta (After establishing that WDR13 expresses in placenta and placenta due to rplacenta . To furt and GCs in Wdr13placenta . Interesplacenta probablyWdr13 mutant placenta: 1) reduced cell proliferation, 2) increased apoptosis and 3) affected EPC, chorion differentiation. So we performed cell proliferation assay in placenta by injecting BrdU in pregnant female mice. At 17.5 dpc there was no difference in number of BrdU positive in Wdr13 mutant and wild type placenta involved in solute transport, angiogenesis and differentiation. There was no difference in glucose and amino acid transporter in Wdr13 mutant and wild type placenta at 17.5 dpc . So we analyzed some AP1 target genes having role in placental trophoblast cell differentiation. We observed that cd44 was down regulated in Wdr13 null placenta as compared to wild type placenta whereas there was no change in axin2, igr5 and c-Jun levels.At least three possibilities exist for the reduction in the number of various trophoblast cells in placenta . Next weplacenta . It appe17.5 dpc . Interesplacenta where asWdr13 null placenta phenotype on postnatal growth of mice. As shown above that XWdr13X heterozygous placenta was deficient in WDR13 protein and XXWdr13 heterozygous placenta showed WDR13 protein levels as comparable to wild type while the two heterozygous female embryos (XWdr13X and XXWdr13) had comparable levels of Wdr13 expression and its high affinity receptor tyrosine kinases VEGFR-1, VEGFR-2 along with angiopoietin-1 (Ang-1), angiopoietin-2 (Ang-2) and its receptor Tunicainterna endothelial cell kinase-2 (Tie-2)212224enotypes . Reducedcd44 in Wdr13 knockout placenta as compared to wild type placenta placenta and embryos. Hemizygous males were crossed with wild type females to generate paternally derived heterozygous (XXWdr13) placenta and embryos. At given time point of pregnancy female mice were dissected and weight of individual placenta and embryos were recorded. Genotyping of individual placenta and embryo was confirmed by isolating DNA from embryo tail as described previouslySry primers (Wdr13X), paternally derived heterozygous (XXWdr13) and wild type females body weight was measured fortnightly on normal diet and on high fat diet.All the mice experiments were approved by and performed under guidelines of the institutional animal ethics committee of CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India. All methods were carried out in accordance with the approved guidelines. dpc and incubated in M16 medium. Same day embryos were transferred to oviduct of pseudo-pregnant female (CD1) after mating with vasectomized male.Isolation and transfer of mouse embryo in oviduct were performed as described previouslyin situ of Wdr13 gene in placenta, primer pairs 5\u2032-aacgcctaccgtacaccaac-3\u2032 and 5\u2032-acatggtacatgcctgcaaa-3\u2032 were used to generate the anti sense and sense probe. Paraffin embedded placenta sections were used to perform RNA in situ using DIG RNA labeling kit from Roche applied sciences (cat no-11175025910) as per manufacturers instructions. To assay the cell proliferation in placenta pregnant female mice were injected BrdU (100\u2009mg/kg body weight), sacrificed 1.5h after injection and fixed and processed as described above. BrdU staining was performed using Anti-BrdU Antibody (Sigma) followed by staining with anti mouse HRP conjugated secondary antibody and developed with DAB substrate . The number of positive cells were counted manually using Axioskop (Axivision software). TUNEL assays were performed using Dead END Fluorometric TUNEL System (Promega). Images were recorded in confocal microscope and TUNEL-positive cells were counted manually from the images from total of 4 mice per genotype.Placentas were collected in PBS, fixed in 4% paraformaldehyde overnight and processed as described previouslyhttp://rsb. info.nih.gov/ij/) on H&E stained samples. A minimum of four placenta for each genotype and at least three sections from each placenta was used to quantify placental area. For counting S-TGCs in labyrinth layer Masson\u2019s trichrome stained placenta were imaged with 1000X magnification. At least four placentas from each genotype were analyzed with three to four field from each placenta. For MBS Masson\u2019s trichrome stained placenta were imaged and ImageJ software was used to calculate MBS per unit area. MBS was calculated from minimum of four placentas per genotype with three to four field per placenta. For determining statistical significance an unpaired two-tailed student\u2019s t-test with unequal variance was used to compare genotypes.Cavalieri technique (Simple grid method) was used to measure area of placenta, JZ, LZ and MBS2dpc placenta was isolated using RNeasy Mini Kit (Qiagen) and reverse transcription was performed using ImProm-IITM kit (Promega) after DNase (Promega) treatment of RNA samples. Real time PCR was performed for various genes using Syber Green master mix (Invitrogen) and list of primers sequences is provided in dpc placentas were lysed in RIPA buffer, quantified and blotted on PVDF membrane. Anti-WDR13 purified antibody (HPA000913) from Sigma and beta actin (sc-47778) from Santacruz were used for visualization of the protein.Total RNA from 17.5 How to cite this article: Singh, V. P. et al. Role of mouse Wdr13 in placental growth; a genetic evidence for lifetime body weight determination by placenta during development. Sci. Rep.5, 13371; doi: 10.1038/srep13371 (2015)."} +{"text": "To compare delayed enhancement DECT with CCTA and LGE MRI for detection of scars after myocardial infarction and to analyze the possible additive value of delayed DECT as part of CCTA protocol.19 patients with history of myocardial infarction ( >1 year) were prospectively enrolled in the study. The CCTA protocol consisted of prospectively gated CTA and DECT. DECT was performed with single-tube 64-row CT in gemstone spectral imaging (GSI) mode with 8 min delay after contrast media injection. Using a 4-point transmurality scale CCTA images were visually assessed for first-pass arterial enhancement deficit and late enhancement in DECT images using iodine distribution maps. Per-segment analysis was performed by 2 observers independently. LGE MRI was performed after CT (range: 1-3 days) as a reference standard. Test characteristics between normal myocardium and scar tissue) for detection of myocardial scar were calculated both for CCTA and DECT. Per segment agreement between modalities was investigated with Spearman rank correlation coefficient.141/323 of LV segments showed LGE on MRI. At segmental level delayed DECT had good accuracy (90%) for scar evaluation with excellent specificity (99%) and satisfactory sensitivity (78%). CCTA protocol without integration of delayed DECT had accuracy, sensitivity and specificity 92%, 88%, 95%, respectively. Addition of delayed DECT results did not improve CCTA performance . CR of scar tissue was higher for CTA 274%\u00b129% vs. 123\u00b16% for DECT, p=0.008). Scores of transmurality were significant lower for CCTA and delayed DECT than for MRI .Detection of ischemic scars with delayed DECT and CCTA showed a good correlation with MRI. Delayed DECT detects myocardial scars with good accuracy but integration of delayed DECT into CCTA protocol did not improved performance of CCTA for detection of chronic scars and could be omitted from cardiac CT protocols in order to reduce radiation exposure to patient.N/A."} +{"text": "Bilaterally absent cortical somatosensory evoked potentials (SSEPs) predict poor outcome after cardiac arrest. A threshold for the amplitude of early cortical SSEPs above which patients may survive with good outcome has not been determined. Thus, tolerable noise levels for the interpretation of cortical SSEPs are poorly defined. Furthermore, it has not been systematically investigated whether high amplitudes of cortical SSEPs may exclude severe hypoxic encephalopathy incompatible with re-awakening.We prospectively studied SSEPs after median nerve stimulation obtained 24 hours to 4 days after cardiac arrest in patients treated with targeted temperature management at 33\u00b0C for 24 hours. Amplitudes of cortical SSEPs were determined, if at least two peripheral, spinal and cortical recordings per side were available, spinal potentials were bilaterally reproducible and cortical noise level was below 0.25 \u03bcV. Cortical SSEP amplitude was defined as largest amplitude of a reproducible cortical SSEP of four cortical recordings (two per side) within 50 milliseconds after stimulation. Outcome was assessed upon ICU discharge using the Cerebral Performance Category (CPC) scale. CPC 1 to 3 was defined as good outcome, CPC 4 to 5 as poor outcome.Of 318 consecutive patients examined, 293 had complete SSEP recordings with reproducible spinal potentials and cortical noise levels below 0.25 \u03bcV. Of those, 137 (47%) had good outcome and 156 (53%) had poor outcome. The lowest amplitude of the early cortical SSEPs in a survivor with good outcome was 0.62 \u03bcV. All 79 patients with amplitudes below this threshold had poor outcome. None of 27 patients who survived with CPC 4 (unresponsive wakefulness syndrome) had cortical SSEP amplitudes above 2.5 \u03bcV. Twenty-four patients with amplitudes above this limit died. Detailed case review indicated a cause of death other than hypoxic encephalopathy in these patients.Our data indicate that the prognostic value of SSEP after cardiac arrest extends beyond a mere absent-present dichotomy. Bilaterally absent as well as very low amplitude SSEPs predict poor outcome with high positive predictive value. SSEPs should not be used for prognostication, if noise in cortical recordings could mask critically low amplitudes. High amplitudes of early cortical SSEPs strongly argue against severe hypoxic encephalopathy incompatible with re-awakening."} +{"text": "To characterize bone metabolism in AIS patients using bone metabolism markers.Although osteopenia is often associated with AIS, bone metabolism in this condition has not been assessed.Bone mineral density (BMD) of the lumbar spine and bilateral proximal femurs and bone metabolism markers {bone formation marker: serum bone alkaline phosphatase (BAP); bone resorption marker: serum tartrate-resistant acid phosphatase serum band 5 (TRAP5b)} were measured in 55 consecutive AIS subjects aged 10 to 18 years-old (mean: 15.6+/-1.7). BMD, body mass index (BMI), and age of menarche were compared between subjects with normal and high values of TRAP5b.Nineteen subjects (34%) had osteopenia and 17 subjects (31%) had osteoporosis. In 51 AIS subjects (93%), values for BAP were within normal range, while 33 subjects (60%) had high values for TRAP5b. Subjects with high values for TRAP5b had BMDs of the lumbar spine significantly lower than BMDs of patients with normal values of TRAP5b.The primary cause of low BMD in AIS was increased bone resorption."} +{"text": "FISH analysis is superior to chromosome analysis in detecting important prognostic genetic abnormalities in PCD. However, its sensitivity is hampered due to paucity of plasma cells in whole bone marrow and often shows false-negative results when frequency of abnormal cells is below the cut-off values. Studies have shown that the abnormality detection rate in enriched plasma cells (EPC) is greater than unselected plasma cells, but purification techniques are limiting to only FISH when bone marrow volumes are inadequate. The inability to perform chromosome analysis may compromise patient care since chromosome analysis is equally important for detecting non-plasma cell related abnormalities, such as secondary myelodysplastic syndrome. To resolve this critical issue and optimize limited quantity received, we designed a study where an immuno-magnetic CD138 enriched positive selection of plasma cells was used for FISH while the negative selection was used to retrieve the remaining cellular components (RCC) for chromosome analysis. After validating this approach in a pilot study, we implemented this strategy in 2012 for routine clinical diagnosis in patients with PCD. When there was adequate sample volume available, both whole bone marrow and RCC were used for chromosome analysis.We found 100% success rate for chromosome analysis using RCC. Identical PCD related genetic abnormalities were observed by both chromosome analysis using whole bone marrow and EPC FISH in 4.2% (4/96) cases while 8.3% (8/96) cases showed discordant results. Population variants such as loss of Y chromosome were observed in 6.3% (6/96) cases. Karyotypic abnormalities of myeloid origin or population variants were found both in whole bone marrow and RCC cultures. Karyotypes with PCD related aberrations were seen only in whole bone marrow cultures in 37.5% of the cases (3/8) while the remaining 62.5% (5/8) of cases showed them in both whole bone marrow and RCC cultures. All PCD related aberrations showed concordant EPC FISH results.Our results confirm the feasibility of retrieving RCC from the CD138 negative fraction, and prove to be an innovative strategy for performing chromosome analysis on PCD patients with insufficient sample volumes."} +{"text": "Ursus maritimus) circumpolar genetic variation during the last two decades of decline in their sea-ice habitat. We sought to evaluate whether their genetic diversity and structure have changed over this period of habitat decline, how their current genetic patterns compare with past patterns, and how genetic demography changed with ancient fluctuations in climate. Characterizing their circumpolar genetic structure using microsatellite data, we defined four clusters that largely correspond to current ecological and oceanographic factors: Eastern Polar Basin, Western Polar Basin, Canadian Archipelago and Southern Canada. We document evidence for recent (ca. last 1\u20133 generations) directional gene flow from Southern Canada and the Eastern Polar Basin towards the Canadian Archipelago, an area hypothesized to be a future refugium for polar bears as climate-induced habitat decline continues. Our data provide empirical evidence in support of this hypothesis. The direction of current gene flow differs from earlier patterns of gene flow in the Holocene. From analyses of mitochondrial DNA, the Canadian Archipelago cluster and the Barents Sea subpopulation within the Eastern Polar Basin cluster did not show signals of population expansion, suggesting these areas may have served also as past interglacial refugia. Mismatch analyses of mitochondrial DNA data from polar and the paraphyletic brown bear (U. arctos) uncovered offset signals in timing of population expansion between the two species, that are attributed to differential demographic responses to past climate cycling. Mitogenomic structure of polar bears was shallow and developed recently, in contrast to the multiple clades of brown bears. We found no genetic signatures of recent hybridization between the species in our large, circumpolar sample, suggesting that recently observed hybrids represent localized events. Documenting changes in subpopulation connectivity will allow polar nations to proactively adjust conservation actions to continuing decline in sea-ice habitat.We provide an expansive analysis of polar bear ( Ursus maritimus), a specialist carnivore whose survival depends on sea-ice for foraging, migration and mating, is expected to be increasingly at risk due to changes in sea-ice habitat The distribution and viability of animal populations are dependent on the quantity, quality and connectivity of habitat. As habitat undergoes change, populations expand or become increasingly isolated. Because habitat change due to climate warming has been and is predicted to be most dramatic at the poles and especially for sea-ice habitat Though movement of individual polar bears across present-day subpopulation boundaries is evident from capture-recapture data and hunter tag returns U. arctos). Both species experienced fluctuations in effective population size throughout their history, including prolonged declines over the past 500,000 years, although this decline is more pronounced in polar bears Microtus spp.) and shrews have suggested that polar bear evolution and effective population size have tracked key climate events, similar to the paraphyletic brown bear were used, unless the accompanying mother and/or sibling were not already in the sample set. Ninety-seven percent of the samples had known latitude and longitude , and 411 samples from 15 subpopulations at the mtDNA control region . Some saongitude ., but anDNA extraction, PCR amplification, and genotyping or sequencing protocols for microsatellite and mtDNA data of ursids are described in the literature We quantified genetic variation and tested for neutrality in both microsatellite and mtDNA data using a variety of computer programs routinely used to analyze genetic data We used microsatellite data to test for differences in the distribution of allele frequencies ST variance based on microsatellite loci were calculated We assessed genetic differentiation among the subpopulations of polar bears using both microsatellite and mtDNA data. Overall standardized estimates of FST values from microsatellite and mtDNA data of each subpopulation, calculated after accounting for differences in the effective size between the two genomes (FST(nu)\u200a=\u200a1\u2013eFST(mt)]0.25*ln and nucleotide diversity [\u03c0]); and coalescent-based simulation methods beauti, part of the v1.6.1 software package Using the mtDNA dataset, we assessed historical signatures of population growth/expansion within subpopulations and/or within larger regional groupings of polar bears using: extended Bayesian skyline plots o; range, 0.64 to 0.73) were similar across subpopulations (o was 0.70. Significant departures from Hardy-Weinberg Equilibrium (HWE) were detected in 5 of 338 cases (locus by subpopulation), with no individual locus or subpopulation accounting for a disproportionate number of significant test results. Accordingly, no loci were dropped from further analyses due to deviation from HWE. Overall, significant (P<0.05) associations of alleles between markers were identified for 47 of 210 pairs of loci (10.5 positive tests would be expected due to Type I error), suggesting that the global population has significant admixture or substructuring. LD patterns were absent with the exception of one pair of loci, suggesting that the observed LD was due to population structure rather than physical linkage on the chromosome. LD within the DS subpopulation, especially in association with, but not isolated to, locus G10X, produced 30 of 67 significant (P<0.002) test results. That the LD was relatively restricted to one subpopulation is suggestive of significant population substructuring within DS.We collected multi-locus microsatellite genotypes from samples of 2,748 polar bears from 18 of 19 circumpolar polar bear subpopulations . Both allelic richness and observed heterozygosity ranged from 0.00 to 0.95 and nucleotide diversity (\u03c0) ranged from 0.000 to 0.007 (Genetic Differentiation of Subpopulations), though most private haplotypes were only represented by a single or few individuals . The upper limit of FST for this data set, taking into account the level of genetic diversity, is 0.295, therefore our overall estimate accounts for 11.5% of the maximum possible level of genetic structure. Thirty-one (20%) of 153 possible pairwise comparisons among subpopulations showed a signal of significant differentiation (adjusted for multiple-comparisons) based on at least one of four metrics . Population pairwise \u03a6ST values ranged from \u22120.046 to 0.805, and 77 of 105 pairwise comparisons were significant in all comparisons that involved subpopulations characterized by three or fewer mtDNA samples . In 33 of 39 comparisons involving these three subpopulations, we rejected the null hypothesis of equal distribution of haplotypes . Among these, the hypothesis that grouped subpopulations into two broad geographic regions yielded the highest mean \u03a6CT value across both marker types and all analyses. In the microsatellite data set, biases in the directionality of gene flow estimated using the coalescent were not as strong as those estimated using allelic frequency ; Fig. 3.Among gene flow estimates based on mtDNA, there was a signal of effective dispersal from the Southern Canada cluster into both Eastern and Western Polar Basin clusters .g\u200a=\u200a915.33, SD\u200a=\u200a48.01), and the mismatch distribution did not deviate significantly from a sudden expansion model but did show significant raggedness . The directional gene flow from the Eastern Polar Basin to the Canadian Archipelago and Western Polar Basin is specifically contemporary , occurring during this current era of sea-ice decline. Estimates of asymmetry in historical gene flow did not show a similar pattern: coalescent estimates of asymmetry in gene flow did not show directionality based on maternally-inherited mtDNA (late Pleistocene signature), or on microsatellite DNA (Holocene signature). The northern parts of the Canadian Archipelago have been predicted to retain polar bear ice habitat farther into the future than other Arctic areas We detected an increase in directional gene flow of polar bears from the Eastern Polar Basin towards the Canadian Archipelago and Western Polar Basin, and from Southern Canada to the Canadian Archipelago, within the last 1\u20133 generations , indicate that demographic fluctuations during the past several generations have been insufficient to influence genetic partitioning of subpopulations within regions.Mismatches between expansions and contractions of polar bears and brown bears appear to coincide with past climate fluctuations. We timed the expansion of Western Polar Basin polar bears to the early Wisconsin glacial period , preceding the expansion of Western Beringian brown bears before the Last Glacial Maximum (LGM), approximately 31,000 years ago, and Eastern Beringian brown bears during the last glacial at \u223c56,000 years ago. Deep nuclear and mitogenomic sequencing attributed to differential responses to climate changes and hybridization The phylogenetic relationships and timing of lineage divergence within and between polar and brown bears remain controversial, due to the difficulty in discerning between incomplete lineage sorting and ancient and recent (periodic) hybridization between the two species The Svalbard region has been proposed as a previous interglacial refugium, retaining a source population for range expansion during cooler periods It is not possible to place radio collars on male polar bears due to the males' neck circumferences exceeding the circumference of the skull, and tracking male polar bears by other means has had mixed success. Limited data have revealed no differences between the male and female movement distances but see . In contThough we increased the geographic sampling of polar bears, our study was limited by lack of modern samples from the Kara Sea in the Russian Arctic (Eastern Polar Basin cluster). This region is of conservation concern due to hydrocarbon exploration and unquantified levels of poaching Our work updates and expands previous circumpolar genetic analyses of polar bears S1 FigLocations of polar bears, a., sampled at known latitude and longitude in 18 circumpolar subpopulations of polar bears, recognized by the IUCN/Polar Bear Specialist Group, and amplified at microsatellite loci: Baffin Bay (BB); Barents Sea (BS); Chukchi Sea (CS); Davis Strait (DS); East Greenland (EG); Foxe Basin (FB); Gulf of Boothia (GB); Kane Basin (KB); Kara Sea (KS); Laptev Sea (LP); Lancaster Sound (LS); M'Clintock Channel (MC); Northern Beaufort Sea (NB); Norwegian Bay (NW); Southern Beaufort Sea (SB); Southern Hudson Bay (SH); Viscount Melville (VM); and Western Hudson Bay (WH). Circles identify bears sampled at known latitude and longitude in the 1980s (n\u200a=\u200a157), 1990s (n\u200a=\u200a613), 2000s and 2010s (n\u200a=\u200a183). b. Locations of 402 polar bears samples in 15 subpopulations with known latitude and longitude amplified at the mitochondrial DNA control region.(TIFF)Click here for additional data file.S2 Figstructure (1), in the circumpolar Arctic.The average of 5 runs per cluster, of the negative log likelihood of the probability of the microsatellite data given the number of clusters of polar bears, K, simulated by the program (TIFF)Click here for additional data file.S3 Figstructure (1): a. two most likely sub-clusters within the Canadian Archipelago Cluster with Baffin Bay (BB) and Davis Strait b. two most likely sub-clusters within the Southern Canada Cluster and c. the two most likely sub-clusters within the Polar Basin: the Eastern Polar Basin Cluster and the Western Polar Basin Cluster .Genetic substructure of polar bears within the three broad clusters identified by program All individuals are sorted left to right by high to low latitude.(TIF)Click here for additional data file.S4 FigST versus pairwise microsatellite Scatter plot of observed pairwise mtDNA \u03a6FST values for 21 microsatellite loci (circles) for 15 subpopulations of polar bears. The line represents the expected microsatellite FST value given the genetic differentiation observed at mtDNA (2): FST(nu)\u200a=\u200a1\u2013eFST(mt)]0.25*ln), Fu's Fs, Tajima's D, raggedness (rg), and Deviation from a sudden expansion (SSD). Bold values for microsatellite data signify microsatellite data from subpopulations that were out of Hardy-Weinberg Equilibrium at \u03b1\u200a=\u200a0.05/number of loci. Bold values for demographic statistics indicate significance at P\u22640.05. Abbreviations of subpopulations are as follows: Baffin Bay (BB); Barents Sea (BS); Chukchi Sea (CS); Davis Strait (DS); East Greenland (EG); Foxe Basin (FB); Gulf of Boothia (GB); Kane Basin (KB); Kara Sea (KS); Laptev Sea (LP); Lancaster Sound (LS); M'Clintock Channel (MC); Northern Beaufort Sea (NB); Norwegian Bay (NW); Southern Beaufort Sea (SB); Southern Hudson Bay (SH); Viscount Melville (VM); and Western Hudson Bay (WH).(DOCX)Click here for additional data file.S2 TableDistribution of haplotypes (and GenBank accession numbers) at the mitochondrial DNA control region in 18 subpopulations of polar bears: Baffin Bay (BB); Barents Sea (BS); Chukchi Sea (CS); Davis Strait (DS); East Greenland (EG); Foxe Basin (FB); Gulf of Boothia (GB); Kane Basin (KB); Kara Sea (KS); Laptev Sea (LP); Lancaster Sound (LS); M'Clintock Channel (MC); Northern Beaufort Sea (NB); Norwegian Bay (NW); Southern Beaufort Sea (SB); Southern Hudson Bay (SH); Viscount Melville (VM); Western Hudson Bay (WH).(DOCX)Click here for additional data file.S3 TableGenetic differentiation results of comparisons of microsatellite data from earlier and later samples within nine global subpopulations and regions of polar bears: the Svalbard portion of the Barents Sea (BS); Baffin Bay (BB); Chukchi Sea (CS); Foxe Basin (FB); Gulf of Boothia (GB); the Labrador portion of Davis Strait (DS); Lancaster Sound (LS); Southern Beaufort Sea (SB) and Western Hudson Bay (WH). Degrees of freedom are shown in parentheses. The K metric represents the likely number of clusters for the subpopulation or region with decadal data pooled as ascertained using the Bayesian clustering program baps. Values in bold show significant differentiation between the groups .(DOCX)Click here for additional data file.S4 TableST, and genotypic differentiation (the latter only for comparisons of subpopulations that were found to be out of Hardy-Weinberg equilibrium) for pairwise comparisons of 18 subpopulations of polar bears using microsatellite data.Below the diagonal, significant (+) and non-significant (\u2212) differences for genic differentiation, F Bonferroni-corrected significance levels were adjusted for the number of loci compared for each pair (0.05/number of loci compared). Shaded blocks delineate the four clusters based on our analysis: Eastern Polar Basin, Western Polar Basin, Canadian Archipelago and Southern Canada. Above the diagonal: significant (+) and non-significant (\u2212) differences for FST, \u03b8ST and haplotypic differentiation using mtDNA data for pairwise comparisons of 15 subpopulations. Abbreviations for subpopulations are as follows: Baffin Bay (BB); Barents Sea (BS); Chukchi Sea (CS); Davis Strait (DS); East Greenland (EG); Foxe Basin (FB); Gulf of Boothia (GB); Kane Basin (KB); Kara Sea (KS); Laptev Sea (LP); Lancaster Sound (LS); M'Clintock Channel (MC); Northern Beaufort Sea (NB); Norwegian Bay (NW); Southern Beaufort Sea (SB); Southern Hudson Bay (SH); Viscount Melville (VM); and Western Hudson Bay (WH).(DOCX)Click here for additional data file.S5 TablePairwise estimates of population differentiation among 15 or 18 (microsatellite DNA) circumpolar subpopulations of polar bears: Baffin Bay (BB); Barents Sea (BS); Chukchi Sea (CS); Davis Strait (DS); East Greenland (EG); Foxe Basin (FB); Gulf of Boothia (GB); Kane Basin (KB); Kara Sea (KS); Laptev Sea (LP); Lancaster Sound (LS); M'Clintock Channel (MC); Northern Beaufort Sea (NB); Norwegian Bay (NW); Southern Beaufort Sea (SB); Southern Hudson Bay (SH); Viscount Melville (VM); Western Hudson Bay (WH). Significant values are in bold text.(DOCX)Click here for additional data file.S6 TableGenetic differentiation within the Southern Hudson Bay (SH) and Davis Strait (DS) subpopulations of polar bears sampled during autumn while bears are on land. In the two-way comparison within DS: Northern indicates samples from polar bears north of Hudson Strait; Southern indicates samples south of Hudson Strait; samples from the Hudson Strait portion of the Foxe Basin subpopulation are not included in this comparison. In the three-way comparison: Southern, as above; Northern Baffin includes samples north of Frobisher Bay on Baffin Island to the border with the Baffin Bay subpopulation; Hudson Strait includes samples south of Frobisher Bay on Baffin Island and along Hudson Strait, including adjacent Hudson Strait samples from the Foxe Basin subpopulation. Akimiski Island is in James Bay, which is in the SH subpopulation; the Hudson Bay Coast is also within SH. All values are significant at \u03b1\u200a=\u200a0.05/number of loci.(DOCX)Click here for additional data file.S7 TableCT, \u03b8SC) and mitochondrial alleles to test hypotheses of groupings of the global subpopulations of polar bears: Baffin Bay (BB); Barents Sea (BS); Chukchi Sea (CS); Davis Strait (DS); East Greenland (EG); Foxe Basin (FB); Gulf of Boothia (GB); Kane Basin (KB); Kara Sea (KS); Laptev Sea (LP); Lancaster Sound (LS); M'Clintock Channel (MC); Northern Beaufort Sea (NB); Norwegian Bay (NW); Southern Beaufort Sea (SB); Southern Hudson Bay (SH); Viscount Melville (VM); and Western Hudson Bay (WH).Hierarchical analysis molecular variance (AMOVA) among subpopulations and various groupings of microsatellite or 0.05 (for mtDNA data).(DOCX)Click here for additional data file.S8 Tablebayesass (1).The proportion of non-migrants (95% CI) over the last ca. 1\u20133 generations for four genetic clusters of polar bears calculated using program The Eastern Polar Basin Cluster includes polar bears from East Greenland, Barents Sea, Kara Sea and Laptev Sea subpopulations. The Western Polar Basin Cluster includes polar bears from the Chukchi Sea, Southern Beaufort Sea and Northern Beaufort Sea subpopulations. The Canadian Archipelago Cluster includes Viscount Melville, M'Clintock Channel, Gulf of Boothia, Lancaster Sound, Norwegian Bay, Kane Basin, Baffin Bay MUs, and the region north of Hudson Strait in the Davis Strait subpopulations. The Southern Canada Cluster includes Foxe Basin, Southern Hudson Bay, Western Hudson Bay MUs, and the region south of Hudson Bay in the Davis Strait subpopulations. Theta (\u03b8) for each cluster is calculated from microsatellite (effective population size (Ne), scaled to mutation rate (\u00b5) and mtDNA data , scaled to mutation rate) using the program MIGRATE.(DOCX)Click here for additional data file.S9 TableCoalescent times (thousands of years ago) to most recent common ancestor for major mitochondrial lineages of brown and polar bears based on 581 bp of the mitochondrial DNA control region, excluding indels. Provided are nodal support for lineages based on Bayesian analysis in BEAST and median age of nodes with 95% confidence intervals (CI). ABC bears refer to brown bears from the Alexander Archipelago, Alaska, USA.(DOCX)Click here for additional data file.S10 TableAverage pairwise distances within and among haplotypes from polar bears sampled from 15 subpopulations, the ancient Poolypenten (GenBank Accession No. GU573488)* polar bear and haplotypes found within the three clades of Alaskan brown bears. Values were generated using the Tamura-Nei (I+G0.69) model of substitution.(DOCX)Click here for additional data file.S11 TableMetadata associated with samples of polar bears collected for microsatellite and mitochondrial DNA (mtDNA) analysis.Table in MS Excel Format.(XLSX)Click here for additional data file.S12 TablePermissions and permits for collection of tissue samples of polar bears used in this study.(DOCX)Click here for additional data file.S1 Supporting InformationDetailed materials and methods.(DOCX)Click here for additional data file."} +{"text": "Haemorrhagic shock (HS)-induced hypotension impairs cerebral perfusion pressure and decreases oxygen supply to the brain (1), but the consequences for brain mitochondrial function are poorly understood.in vivo.To investigate the effects of HS on mitochondrial function in the cerebral cortex of rats n=7) compared with sham (n=6) controls. Mitochondrial function was assessed by observing mitochondrial membrane potential (using tetramethyl rhodamine methyl ester (TMRM); ex: 543 nm; em: 585 nm), and endogenous flavin adenine dinucleotide (FAD) autofluorescence by confocal imaging. The TMRM and FAD signals were registered before induction of HS (baseline), at MAP of 40 mmHg (shock), and at 30 (T30) and 60 (T60) minutes after shock.A flap of skull was removed and cerebral cortex exposed in isoflurane-anaesthetised rats undergoing HS of 40 mmHg for 30 minutes) (p < 0.001) and reduction of the FAD/FADH2 pair at both T30 and T60 post-shock, except for a striking protection observed as a 'halo' extending ~30-40 mm from arterioles (Figure There was a loss of mitochondrial membrane potential (decreased TMRM signal; HS-induced hypotension causes a marked loss of mitochondrial membrane potential and FAD fluorescence in cerebral cortex, except in regions in immediate proximity to arterioles. This study reveals a profound spatial vulnerability of cortical tissue to reduced blood flow."} +{"text": "Department of Radiology is one of the intermediate cost centres for supporting patient treatment in wards and clinics in Universiti Kebangsaan Malaysia Medical Centre (UKMMC). A total of 121,221 procedures were performed in 2011. Unfortunately, since UKMMC began its operations, the actual cost of each procedure in this particular department has never been studied. Hence, it is important to determine the actual cost of each procedure in preparation for future autonomous university governance, particularly budgeting.A cross-sectional study was conducted from January to December 2012 in all units in Department of Radiology. Activity Based Costing (ABC) methodology was used to analyse the cost of each procedure. Three hundred procedures had been collected using a micro costing form. Seven cost components were collected for each procedure: (1) staff salary (2) consumables (3) equipment (4) reagents (5) administration (6) maintenance (7) utilities. The results were analysed using SPSS version 22.0.The five highest costs of radiological procedures had been identified. The highest mean cost procedures were Endovascular Interventional Radiology (EIR) (RM2214\u00b11636), followed by Medical Nuclear (RM1741\u00b1 908), Magnetic Resonance Imaging (MRI) (RM1323\u00b1232), Computed Tomography (CT Scan) (RM395\u00b1152) and Mammogram (RM322\u00b14). In addition, the highest component cost for EIR procedure were consumables , for Medical Nuclear and MRI procedures, cost of equipment was the highest component which was RM875 (50%) and RM797 (60%), respectively. For CT scan procedures, cost of reagents was the highest component , which was followed by cost of consumables for Mammogram .An accurate and precise cost per radiological procedure can be determined by using ABC method. The important component that contributes to each of the activities performed at each procedure can be further justified."} +{"text": "ECG revealed sinus rhythmwith complete right bundle brunch block. Transthoracic echocardiography demonstrated asecundum atrial septal defect (ASD) and Qp/Qs ratio was 1.6. The right ventricle wasseverely dilated, inconsistent with the size of the defect. Two-dimensional and colorDoppler transesophageal echocardiographic examination confirmed the secundum type ASD(asterisk) and revealed an additional sinus venosus type ASD between the right atriumand the superior vena cava (SVC); Video 1ava (SVC). The pat1 In patients withseverely dilated right ventricle and high Qp/Qs ratio, inconsistent with defect size,physicians should consider the presence of additional septal defects. These patientsmust be evaluated with advanced imaging modalities.The interatrial septum is anatomically divided into 5 septal zones. A mixed atrial septaldefect involves 2 or more of the 5 septal zones and accounts for 7 of all atrial septaldefects."} +{"text": "Occupational health surveillance data are vital to effective intervention. Limited information is available on the magnitude of occupational injuries among laboratory personnel in Kenya.We set out to quantify the prevalence of hazardous injuries among laboratory personnel in Kenya.As part of the Kenya\u2019s premier national public health laboratory\u2019s training on bio-safety and bio-security, laboratory personnel were invited to take part in a survey on occupational hazards and the safety climate at their workstations. Descriptive statistics were used to summarize types of hazardous incidents experienced by laboratory personnel. Logistic regression was used to describe factors associated with reporting a hazardous injury.One hundred and sixteen laboratory personnel drawn from 108 heath facilities participated. Majority were drawn from public health facilities (90%); the others were from private health facilities (8%) and faith based organizations (2%). Twenty-five (22%) were from facilities that had reporting mechanisms for occupational exposures. The median duration of service was 4 years (Range 0.2-33.0) and 18 (16%) had ever been trained on bio-safety. Eighty-nine (77%) personnel experienced by 127 incidents, these were: spills (46), sharps injuries (38), hazardous gas inhalation (19), subcutaneous chemical exposures (17), falls (6) and hazardous agents ingestion (1). Incidents occurred during spillage (44%), laboratory procedures (35%), waste handling (28%), surface-contamination (22%), maintaining equipment (9%), device use (7%),while others were due to inappropriate dressing (8%), food stuff in work area (4%), fires (2%) and heavy lifting (1%). At the time of incident, PPE donned by the majority were gloves (87%) and lab coats (82%). Only 63% (56) reported their incidents; sharp injuries were more likely to have been reported .Due to the magnitude of occupational hazards, an integrated information and incident management system should be implemented to routinely document occupational hazards.None declared."} +{"text": "Respiratory syncytial virus (RSV) is a major cause of community-acquired severe respiratory illness in infants, immunocompromised individuals and the elderly. Limited information exists on healthcare associated RSV infections in developing countries.To describe hospital-acquired RSV infections in three Kenyan referral hospitalsOngoing surveillance for healthcare associated infections is conducted at three referral hospitals in Nairobi: Kenyatta National Hospital (KNH), Jaramogi Oginga Odinga Teaching and Referral Hospital (JOOTRH) and Mbagathi District Hospital (MDH). We collected nasopharyngeal and oropharyngeal samples from patients with new-onset fever (\u226538\u00b0C) and either cough or sore throat, after being afebrile for at least three days in the wards. Specimen were tested for RSV using real time polymerase chain reaction (RT-PCR) and those positive with a cycle threshold value of 30 and below were further grouped as RSV A or B using the same method. The ectodomain of the attachment G glycoprotein was sequenced and phylogenetically analyzed.Among 255 cases tested from September, 2009 to September, 2011, 37 (14.5%) were positive for RSV, including 13 (35%) subgroup A, 6 (16%) B, 1 (3%) mixed AB and 17 (46%) could not be determined. Seventeen samples were successfully sequenced out of the twenty samples on which this was attempted. Majority of our RSV A isolates belonged to NA1 genotype prototype strain and all RSV B sequences clustered with the BAIV genotype. Three RSV A and 2 RSV B sequences from patients on the same ward at KNH were 100% identical in the G ectodomain suggesting potential common source. One RSV A positive specimen from MDH and one from JOOTRH showed 100% sequence identity.Presence of identical sequences indicates potential patient to patient transmission of RSV within the hospitals. Effective and feasible infection control strategies should be enhanced in the Kenyan public hospitals.None declared."} +{"text": "Products secreted from nerve terminals can have profound effects on the phenotype and functioning of T cells. One such product, vasoactive intestinal peptide (VIP) is a small neuroendocrine peptide hormone with potent anti-inflammatory activities. Binding to either of its type B G protein coupled receptors VPAC1 and VPAC2 initiates a signaling cascade that ultimately inhibits the secretion of pro-inflammatory cytokines and reduces proliferative capacity via inhibition of NF-\u03baB["} +{"text": "In contrast, no changes were observed in cardiac vagal tone and bradycardic response to both baroreflex and chemoreflex between LAD and Sham groups. The immediate sympathoexcitatory response in LAD rats was dependent on an excitatory spinal sympathetic cardiocardiac reflex, whereas at 3 h an angiotensin II type 1 receptor mechanism was essential since Losartan curbed the response by 34% relative to LAD rats administered saline (P<0.05). A spinal reflex appears key to the immediate sympathoexcitatory response after coronary ligation. Therefore, the sympathoexcitatory response seems to be maintained by an angiotensinergic mechanism and concomitant augmentation of sympathoexcitatory reflexes.Our aim was to assess the timing and mechanisms of the sympathoexcitation that occurs immediately after coronary ligation. We recorded thoracic sympathetic (tSNA) and phrenic activities, heart rate (HR) and perfusion pressure in Wistar rats subjected to either ligation of the left anterior descending coronary artery (LAD) or Sham operated in the working heart-brainstem preparation. Thirty minutes after LAD ligation, tSNA had increased , being even higher at 60 min ; while no change was observed in Sham animals. HR increased significantly 45 min after LAD (P<0.01). Sixty minutes after LAD ligation, there was: (i) an augmented peripheral chemoreflex \u2013 greater sympathoexcitatory response ; (ii) an elevated pressor response and a reduced baroreflex sympathetic gain (1.3\u00b10.1 versus Sham 2.0\u00b10.1%.mmHg Coronary artery ligation is accepted to mimic myocardial infarction (MI) causing sympathetic nervous system over activity Most of our knowledge on changes in cardiovascular autonomic balance has been obtained days after MI or later in the course of chronic heart failure, using indirect methodological approaches such as measuring plasma catecholamine levels Experiments were performed on male Wistar rats (60\u2013100 g) using an anaesthetic-free preparation with a preserved functional brainstem \u2013 the working heart-brainstem preparation (WHBP) 2\u22125% CO2), until loss of the paw withdrawal reflex. Following sub-diaphragmatic transection, rats were exsanguinated and anaesthesia terminated. Preparations were submerged in ice-chilled Ringer solution , decerebrated pre-collicularly and a 6.0 suture passed around LAD. The preparation was placed into a recording chamber and a double-lumen catheter inserted into the descending aorta for retrograde perfusion. The second lumen of the catheter was used to monitor perfusion pressure (PP). The perfusate was carbogen gassed (to pH 7.4), warmed {31\u00b0C; which was optimal for viability as originally described by Paton Animals were pre-treated with heparin sodium and deeply anaesthetized with isoflurane and thoracic sympathetic nerve activities (tSNA), HR, ECG and PP, either LAD ligation was carefully performed without affecting nerve recordings; or the suture remained untied in Sham operated preparations. At the end of each experiment, the heart was rapidly excised and via the ascending aorta perfused with 1% Evans blue dye (Sigma) to indicate the myocardial area at risk as reported previously \u22121). The percentage of sympathoinhibition was obtained from the ratio of mean tSNA during the peak of the PP increase against the mean activity from a preceding equivalent time period, at an equivalent stage of the respiratory cycle. The bradycardic baroreflex gain was quantified as \u0394HR/\u0394pressure (bpm.mmHg\u22121). (III) Cardiac sympathetic tone was assessed by adding atenolol to the perfusate to block cardiac \u03b21-adrenoceptors and inferred from the HR change. (IV) Bilateral cervical vagotomy was performed to measure the cardiac vagal tone as expressed by the HR change. (V) Angiotensin II type 1 receptor (AT1R) antagonist, Losartan potassium (Sigma), was used to evaluate the role of an AT1R in the initiation and maintenance of sympathetic activity post LAD ligation. Either Losartan or saline was added to the perfusate 1 h and re-applied 2 h post LAD ligation and tSNA recorded up to 3 h after LAD ligation. Half the dose of Losartan used in this study is able to block angiotensin II effects in the mediation of the baroreceptor reflex in the nucleus of the solitary tract Our aim was to examine changes in respiratory and cardiovascular autonomic tone and its reflex control immediately after coronary ligation while HR increased by 45 min [The sympathoexcitatory component of the response to peripheral chemoreceptor activation was greater 60 min post LAD ligation compared to Sham . The baroreflex bradycardia was unaffected after LAD ligation (Both the pressor effect and the sympathetic (non-cardiac) baroreflex gain evoked by phenylephrine were significantly different 60 min after LAD ligation versus Sham rats after atenolol. Cardiac vagal tone was unaffected by LAD ligation . The level of mean tSNA was also not attenuated by bilateral vagotomy , ruling out a vagal mechanism for sympathoexcitation post LAD ligation.We assessed whether AT1R were important for the sympathoexcitation post LAD ligation. The mean level of tSNA became different from 3 h onwards (33.8% of difference) between Losartan (n\u200a=\u200a10) and Control (n\u200a=\u200a9) LAD groups , inspiration (I), post-inspiration (PI) and mid-expiration (Mid E) 30 and 60 min after LAD ligation (n\u200a=\u200a21) and in Sham (n\u200a=\u200a19), we observed that tSNA was higher in all expiratory and inspiratory (I) phases in the LAD group [P<0.01 compared to basal; see The chronic over active sympathetic nervous system post coronary artery ligation has been intensively researched. In contrast, the acute responses of the autonomic nervous system, their onset and mechanisms following MI remain to be well clarified. In the last decades, studies have reported sudden (in the first minute) cardiac afferent and efferent sympathetic responses to transient ischaemia via short-term coronary occlusion in anaesthetized cats Most experimental studies in humans and animals models have used indirect methods/indices to provide information about changes in the balance of autonomic activity after MI; this reflects the technical difficulty of obtaining direct measurements of autonomic nervous activity. Some studies have investigated plasma catecholamine levels Graham et al. In accordance with the increase observed in basal sympathetic activity, the sympathetic components of two major homeostatic reflexes were also altered within 60 min post LAD ligation. We observed a greater chemoreflex evoked sympathoexcitation and depressed sympathetic (non-cardiac) baroreflex gain (i.e. impaired sympathoinhibition), suggesting up-modulation of pro-sympathetic reflex pathways, which is consistent with studies where renal sympathetic nerve activity was measured in pacing-induced chronic heart failure rabbits In contrast to widespread sympathoactivation following LAD ligation, we found that both baseline cardiac vagal tone and reflex bradycardia evoked from both the peripheral chemo- and baro- receptors were unaffected. Variable information is available regarding both tonic and reflex cardiac parasympathetic modulation immediately post infarction. Some studies in humans and cats suggest cardiac vagal over activity in the first minutes/hours following MI The role of the brain renin-angiotensin II system (RAS) in modulating sympathetic outflow and baroreflex sensitivity Our data revealed that an angiotensinergic mechanism mediated via AT1R underpins the elevation in sympathetic outflow which is functionally detected from 3 h post LAD ligation. Evidence reveals AT1R located in circumventricular organs , hypothalamic paraventricular nucleus and the supraoptic nucleus are activated following MI and cause sympathoexcitation The initiating mechanism triggering sympathetic over activity after cardiac ischaemia in rats may include a spinal sympathetic cardiocardiac reflex. This reflex was firstly reported by Malliani et al. Besides the cardiac sympathetic afferents, arterial baroreceptor reflex may be altered simultaneously during myocardial ischaemia, because of their unloading triggering reflex increases in efferent sympathetic nerve activity The present findings indicate selective up-regulation of sympathetic activity with no change in cardiac vagal tone or reflex modulation. It is reasonable to assume that this initial modulation leads to a protective effect to the heart supporting cardiac function In summary, our work shows both increased basal sympathetic drive and chemoreflex sympathoactivation but no changes in vagal regulation in the first hour following coronary artery ligation. Both spinal cord mechanisms and later elevated angiotensinergic activity contribute to increased sympathetic activity generation, some of which is directed to the heart. A detailed knowledge of any mechanisms that are responsible for the autonomic responses following cardiac ischaemia may assist in the temporal design of future effective pharmacological interventions.Figure S1HR changes post LAD ligation in spinal animals. Means of HR before and after spinal cord transection, and in the following 15, 30, 45 and 60 min post LAD ligation and in Sham operated . **P<0.01, *P<0.05 and ##P<0.01.(TIF)Click here for additional data file.Figure S2Respiratory-sympathetic activity coupling. Phrenic-triggered mean integrated tSNA from four respiratory phases \u2013 late expiration (Late E), inspiration (I), post-inspiration (PI) and mid-expiration (Mid E) \u2013 before and at 30 and 60 min post LAD ligation and in Sham . **, ##, &&, \u2020\u2020P<0.01 compared to respective baseline; the magnitude of increase at 30 and 60 min in relation to baseline is given in percentage.(TIF)Click here for additional data file."} +{"text": "Neurally adjusted ventilatory assist (NAVA) is used to adapt mechanical ventilation to patient demand while unloading respiratory muscles. Titration methods are focused on sustained unloading of the diaphragm while maintaining a stable tidal volume . It is hTo assess the extra-diaphragmatic muscle activity (EDMA) during 100%, 50% and 150% of titrated NAVA level.EDMA was measured in ventilated patients with mild ARDS. EDMA was defined as the amplitude of the combined surface EMG at the scalene and sternomastoid muscle . A baseline NAVA level (NAVA100) was titrated using the diaphragm activity (EAdi) response to changing NAVA levels, according to Brander et al. [Twenty-one patients were included. In six patients EDMA was absent during NAVA100, so NAVA titration was sufficient to unload accessory respiratory muscles. Fifteen patients (71%) showed EDMA during NAVA100. In seven patients (33%) EDMA increased at NAVA50 and decreased during NAVA150. In one patient, EDMA decreased at NAVA150. One patient only showed EDMA during NAVA50. In three patients EDMA decreased at NAVA50 or increased at NAVA150. Nine patients (43%) showed no change in EDMA after a change of NAVA level.Measurement of extra-diaphragmatic muscle activity using surface EMG during NAVA ventilation might be helpful in titration of ventilatory assist level in order to optimize patient\u00b4s comfort."} +{"text": "Suidae speciation and domestication, porcine IFNs evolutionarily engender both molecular and functional diversification, which have not been well addressed in pigs, an important livestock species and animal model for biomedical sciences. Annotation of current swine genome assembly Sscrofa10.2 reveals 57 functional genes and 16 pseudogenes of type I IFNs. Subfamilies of multiple IFNA, IFNW and porcine-specific IFND genes are separated into four clusters with \u223c60 kb intervals within the IFNB/IFNE bordered region in SSC1, and each cluster contains mingled subtypes of IFNA, IFNW and IFND. Further curation of the 57 functional IFN genes indicates that they include 18 potential artifactual duplicates. We performed phylogenetic construction as well as analyses of gene duplication/conversion and natural selection and showed that porcine type I IFN genes have been undergoing active diversification through both gene duplication and conversion. Extensive analyses of the non-coding sequences proximal to all IFN coding regions identified several genomic repetitive elements significantly associated with different IFN subtypes. Family-wide studies further revealed their molecular diversity with respect to differential expression and restrictive activity on the resurgence of a porcine endogenous retrovirus. Based on predicted 3-D structures of representative animal IFNs and inferred activity, we categorized the general functional propensity underlying the structure-activity relationship. Evidence indicates gene expansion of porcine type I IFNs. Genomic repetitive elements that associated with IFN subtypes may serve as molecular signatures of respective IFN subtypes and genomic mechanisms to mediate IFN gene evolution and expression. In summary, the porcine type I IFN profile has been phylogenetically defined family-wide and linked to diverse expression and antiviral activity, which is important information for further biological studies across the porcine type I IFN family.Type I interferons (IFNs), key antiviral cytokines, evolve to adapt with ever-changing viral threats during vertebrate speciation. Due to novel pathogenic pressure associated with Interferons (IFNs) are key cytokines that regulate immune responses against viral infection in vertebrates Mammalian type I IFNs probably emerged during tetrapod evolution from fish Type I IFNs are multifunctional despite their central role in antiviral responses Of porcine type I IFNs, IFN-\u03b1 and IFN-\u03b2 have been studied mostly for their response to and activity during viral infections Our objective was to determine the molecular diversity of porcine type I IFN genes, and to correlate this to their expression and function to porcine development and immunity. Extensive annotation of the current swine genome assembly (Sscrofa10.2) identified a total of 57 type I IFN functional genes plus 16 pseudogenes, clearly indicating gene expansion comparable to that in cattle but much larger than the gene composition of type I IFNs in humans and mice [7], [8]Further examination of the 57 porcine type I IFNs functional genes revealed 18 duplicates, which contain ORFs encoding protein precursors identical to one of the 39 previously defined IFNs. These duplicates belong to the subclasses of IFN-\u03b1, IFN-\u03b4 and IFN-\u03c9 with multiple genes. To examine whether these duplicates are assembly artifacts or authentic porcine IFN paralogs, we extracted the genomic sequence adjoining each ORF for approximately 4 Kb, which flanks 2 Kb 5\u2032-upstream and 1 Kb 3\u2032-downstream from the ORF and tentatively includes major gene elements of promoter and untranslated regions. Despite the high identity of ORF IFNs (xIFN1\u20135) were included in our phylogenic analysis. As shown in Gallus gallus) IFNs (gIFNs) and selected mammalian type I IFNs, primarily porcine types. Notably, zIFNphi1 forms a clade with xIFN3\u20135 and chicken gIFNs, whereas the other two zebrafish IFNs, zIFNphi2 and zIFNphi3, have a closer relationship with amphibian xIFN1 and xIFN2 and, thus, are more ancient than mammalian type I IFNs. Considering that genes of both zebrafish and frog IFN precursors contain several introns, but genes of chicken and mammalian type I IFNs are intronless, the retroposition events leading to loss of introns in IFN genes could have occurred independently in birds or mammals because their type I IFNs evolved from different prototypes Phylogenic analysis of type I IFNs in pigs and other species implies both a paralogous relationship within the porcine subclasses and cross-species orthologous speciation http://www.ncbi.nlm.nih.gov/genome/), we analyzed gene conversion among porcine type I IFN sequences. Gene conversion was analyzed with GENECONV software using the default setting to compute both the Sim P-value (estimated based on permutation) and the Bonferroni-corrected (BC) KA (BLAST-like) P-value In addition to gene duplication proposed as a major mechanism for IFN diversification, gene conversion, which refers to fragmental replacement between homologous sequences , also might be involved in evolution of the type I IFN gene family. Using the ORF-centered 4 Kb-long gene pieces (tentatively including 5\u2032-promoter and 3\u2032-untranslated contexts) extracted from the NCBI genome database are remnants of ancestral retroviral integration into the genome of germ-line cells constituting 4\u201310% of genome sequences in different animal species http://www.ncbi.nlm.nih.gov/protein/). The subtype consensus sequences (IFN-con) within or across species were generated and manually curated to represent most conservative residues among the aligned IFN peptides. The 3-D structures of individual or consensus IFN peptides were modeled based on the structures of closely related homologs in structure databases To reveal the potential structure-activity relationship, we collected approximately 200 representative IFN sequences from lower vertebrates to main mammalian species deposited in the NCBI protein database http://www.ncbi.nlm.nih.gov/gene/) and further curated using BLASTP against the current swine genome assembly (Sscrofa10.2) Annotation was done primarily with Otterlace/Zmap manual annotation software via web-based interaction with the Wellcome Trust Sanger Institute (WTSI) SingleSignOn system (affiliated with the Immune Response Annotation Group (IRAG) of the swine genome project) As previously defined, genes that yielded a one:many relationship during the orthology search were subjected to an additional round of similarity analysis. Artifactual duplication status was designated when genes possessed approximately 99% identity at the nucleotide level and were in a cluster of proximal genes tandemly duplicated at the same level of identity Gene conversion was analyzed with GENECONV http://services.cbu.uib.no/tools/kaks) for calculation of synonymous (dS) or non-synonoymous (dN) changes in the coding regions, and the dS and dN values were plotted against one another. Significant estimation (p<0.05) of positive and purifying selections were also calculated using MEGA5 Each gene pair in the IFN gene subfamilies containing multiple function genes were analyzed pairwise using a dN/dS calculation tool (http://www.repeatmasker.org/) in default mode. The IFN sub-locus association of repetitive elements was examined with the Censor program http://www.mas.ncl.ac.uk/~ntmwn/compare2trees) About 1.1 Mbp region in Chromosome (SSC) 1 containing the porcine type I IFN locus was extracted from NCBI Reference Sequence NC_010443.4, and repetitive elements within were determined using the RepeatMasker program and their associated repetitive elements. The phylogenies of Newick strings of both IFN genes and associated repetitive elements were generated using the MEGA http://www.mas.ncl.ac.uk/~ntmwn/compare2trees). The overall topological scores (at the bottom of each comparison) between the IFN genes and allied REs were reported in (PDF)Click here for additional data file.Table S1Repetitive elements (RE) associated with the genes of porcine type I IFN subtypes. The genomic sequence was extracted from chromosome 1 (SSC1) contig NC_010443.4 (http://www.ncbi.nlm.nih.gov/nuccore/NC_010443.4|:224375846\u2013225698870). The association of repetitive elements (REs) was examined with the Censor program http://www.girinst.org/censor/index.php) to screen the 1.3 Mbp sequence on SSC1, which contains all porcine Type I IFN genes except the IFNK gene. The program reported the name, class, and length of the detected REs. The \u201csimilarity value (Sim)\u201d reports the number of matches in the alignment normalized by the length of the alignment, whereas the \u201cpositive value (Pos)\u201d reports the normalized number of alignment positions that produce positive scores in the alignment matrix. The location of either REs or IFN genes (ORFs) is related to the nt positions in the 1.3 Mbp region, and \u201cdistance to IFN ORF\u201d was obtained to subtract the start position of a IFN ORF from the position of the closer end of the corresponding RE(s); the negative numbers indicate REs posited prior to 5\u2032-end of the ORFs, and positive numbers for that after 3\u2032-end of the ORFs. The IFN genes and corresponding REs are organized according to the order localized on SSC1, as shown in (XLSX)Click here for additional data file.Table S2Gene recombination events deteted by the RDP3 package In brief, multiple sequence alignment files generated from the MUSCLE (XLSX)Click here for additional data file."} +{"text": "In vivo, plasma from mice anesthetized with isoflurane had significantly higher endothelial cell-derived CD144+ CD73+ microparticles and had increased microparticle CD73 activity compared to plasma from pentobarbital-anesthetized mice. Supporting a critical role of CD73 in isoflurane-mediated endothelial protection, a selective CD73 inhibitor (APCP) prevented isoflurane-induced protection against human endothelial cell inflammation and apoptosis. In addition, isoflurane activated endothelial cells Rho kinase evidenced by myosin phosphatase target subunit-1 and myosin light chain phosphorylation. Furthermore, isoflurane-induced release of CD73 containing microparticles was significantly attenuated by a selective Rho kinase inhibitor (Y27632). Taken together, we conclude that the volatile anesthetic isoflurane causes Rho kinase-mediated release of endothelial microparticles containing preformed CD73 and increase adenosine generation to protect against endothelial apoptosis and inflammation.Endothelial dysfunction is common in acute and chronic organ injury. Isoflurane is a widely used halogenated volatile anesthetic during the perioperative period and protects against endothelial cell death and inflammation. In this study, we tested whether isoflurane induces endothelial ecto-5\u2032-nucleotidase (CD73) and cytoprotective adenosine generation to protect against endothelial cell injury. Clinically relevant concentrations of isoflurane induced CD73 activity and increased adenosine generation in cultured human umbilical vein or mouse glomerular endothelial cells. Surprisingly, isoflurane-mediated induction of endothelial CD73 activity occurred within 1 hr and without synthesizing new CD73. We determined that isoflurane rapidly increased CD73 containing endothelial microparticles into the cell culture media. Indeed, microparticles isolated from isoflurane-treated endothelial cells had significantly higher CD73 activity as well as increased CD73 protein. More than 1 trillion endothelial cells in human body cover the entire circulatory system and play an integral role in maintaining homeostasis of all organs Volatile anesthetic is administered to virtually all patients subjected to general anesthesia making it one of the most frequently used medications during the perioperative period After inhalation, volatile anesthetics are first taken up by the pulmonary circulatory system and all endothelial cells in the body are rapidly exposed. We and others previously demonstrated that volatile anesthetics protect against endothelial cell necrosis, apoptosis and inflammation due to hypoxia, lipopolysaccharide and cytokine exposure All animal work was approved by Columbia University Institutional Animal Care and Use Committee.Immortalized human umbilical vein endothelial cells cells were grown in high-glucose DMEM plus 10% FBS. Immortalized mouse glomerular endothelial cells (GENC) were obtained from Dr. M. Madaio (Georgia Regents University) and grown in low glucose DMEM/Ham\u2019s F12 medium plus 10% FBS, 2 mM l-glutamine, and 10 mM HEPES Endothelial cells were exposed to isoflurane as described previously After Columbia University Institutional Animal Care and Use Committee approval, we anesthetized adult male C57BL/6 to 4 hr of equipotent doses of either pentobarbital or isoflurane (1.2% or \u223c1 MAC) as described previously et al. Microparticles were isolated by differential centrifugation as described by Amabile We also subjected isolated endothelial and mouse plasma microparticles to flow cytometric analysis with a Quanta SC flow cytometer . To determine the relative expression of CD73, microparticles isolated from endothelial cell culture media were stained with Annexin V and CD73 for 5 min at room temperature. Mouse plasma microparticles were stained with CD73 and CD144 to determine the CD73 expression in endothelial microparticles.After exposure to 2.5% isoflurane or with carrier gas for 1 hr, EA.hy9262 cells were exposed to tumor necrosis factor-alpha plus cycloheximide (10 \u00b5g/ml) for 16 hr to induce apoptosis or to TNF-\u03b1 (20 ng/ml) for 6 hr to induce inflammation as described previously We measured mRNA encoding human CD73 after isoflurane treatment as described EA.hy9262 or mouse glomerular endothelial cell culture media were collected after isoflurane treatment and assayed for adenosine by HPLC as described et al. CD73 activity was measured by tracking the conversion of AMP to adenosine with or without 100 \u00b5M APCP using a modified protocol according to Gelain After 30 min treatment with 2.5% isoflurane or with carrier gas, we measured EA.hy9262 cell Rho kinase activity with a commercial assay that measures myosin phosphatase target subunit-1 (MYPT-1) Threonine residue 696 phosphorylation . We also assessed Rho kinase activation by detecting myosin light chain (MLC) Serine-19 phosphorylation. EA.hy9262 cell lysates were probed with anti-phosphor-MLC 2 antibody as well as total-MLC 2 antibody and subjected to immunoblotting as described The data were analyzed with Student\u2019s t-test when comparing means between 2 groups or with one way analysis of variance plus TUKEY\u2019s post hoc multiple comparison test to compare mean values across multiple treatment groups. All data are expressed throughout the text as means \u00b1 SEM.2) cells or mouse glomerular endothelial cells (GENC) were treated with 0\u20132.5% isoflurane for 6 hr and we measured adenosine levels in cell culture media with HPLC as described previously 2) cells .The next set of experiments determined whether isoflurane stimulates CD73 activity to increase adenosine generation in cultured endothelial cells. EA.hy9262 cells had significantly increased cell surface CD73 activity (conversion of AMP to adenosine) 1 to 3 hr after treatment with 2.5% isoflurane . We alsoSince isoflurane transiently increases cell plasma membrane CD73 activity without inducing new CD73 synthesis in cultured endothelial cells, we tested whether isoflurane induces the release of plasma membrane microparticles containing CD73. Indeed, microparticles isolated from cell culture media of EA.hy9262 cells treated with 1.25 or 2.5% isoflurane for 1 hr had significantly increased CD73 activity compared to microparticles isolated from carrier gas-treated cells . IsoflurWe next determined whether isoflurane anesthesia increases endothelial cell-derived CD73 containing microparticles in plasma of mice anesthetized with isoflurane. We then tested whether CD73 is critical for isoflurane-mediated protection against endothelial apoptosis and inflammation. EA.hy9262 cells pretreated with carrier gas underwent apoptosis with robust PARP and caspase-3 fragmentation after TNSince previous studies showed that Rho kinase activation increases endothelial microparticle generation Overactive inflammatory response is a detrimental complication of surgery and perioperative surgical infections. Anti-inflammatory and cytoprotective effects of volatile inhalational anesthetics are well recognized in vivo and in vitroIn addition to its analgesic and anesthetic properties, volatile anesthetics have non-anesthetic effects in virtually every cell type. Importantly, volatile anesthetics protect against cell death and inflammation in several key organs including the heart, brain, kidney and intestine. For example, several clinically utilized volatile anesthetics including isoflurane precondition the heart against ischemia and reperfusion injury in vivo attenuates systemic cytokine (IL-1\u03b2 and IL-6) upregulation and well as lung injury in vitroin vivo and in vitro, the detained cytoprotective mechanisms remained elusive.Previous studies have shown that isoflurane anesthesia in mice and rats attenuates lipopolysaccharide induced endothelial inflammation 1, A2a, A2b and A3 adenosine receptors] 1, A2a or A2bARs protects against ischemia reperfusion injury in the kidney, heart, liver and brain We recently showed that isoflurane protected against renal tubular necrosis, apoptosis and inflammation by direct induction of CD73 enzyme and activity leading to enhanced adenosine generation In this study, we demonstrate that isoflurane rapidly released endothelial microparticles containing preformed CD73 in cultured endothelial cells as well as in plasma of mice. Indeed, CD73 was directly responsible for isoflurane-mediated endothelial cell protection. Plasma membrane microparticles are phospholipid microvesicles of submicron (0.1 to 1.0 \u00b5m) fragments that originate from plasma membrane blebbing and are subsequently shed et alWe also demonstrate an important role for Rho kinase activation in isoflurane-mediated CD73 containing endothelial microparticle release. We show in this study that isoflurane-mediated endothelial microparticle release and induction of CD73 activity were significantly attenuated by a selective Rho kinase inhibitor. In many cell types including endothelial cells, Rho kinase regulates cytoskeleton architecture, migration and growth In summary, we demonstrate that a commonly utilized volatile anesthetic isoflurane rapidly increases endothelial cell adenosine generation via releasing microparticles containing preformed CD73. Release of CD73 and subsequent adenosine generation may result in cellular protection in neighboring and remote endothelial and epithelial cells via activation of adenosine receptors. Our current findings in endothelial cells differ considerably from findings in renal tubular epithelial cells as increased adenosine generation occurred without the induction of new CD73 synthesis. Taken together, our current study provides additional mechanistic insight into the mechanism of isoflurane-mediated endothelial cell protection."} +{"text": "Investigation of metabolic responses to high night temperatures in 12 differently sensitive rice cultivars showed strong effects on central metabolism in sensitive and metabolic pre-adaptation in tolerant cultivars. Oryza sativa L.). Little is known about metabolic responses of rice to high night temperature (HNT) conditions. Twelve cultivars with different HNT sensitivity were used to investigate metabolic changes in the vegetative stage under HNT compared to control conditions. Central metabolism, especially TCA cycle and amino acid biosynthesis, were strongly affected particularly in sensitive cultivars. Levels of several metabolites were correlated with HNT sensitivity. Furthermore, pool sizes of some metabolites negatively correlated with HNT sensitivity under control conditions, indicating metabolic pre-adaptation in tolerant cultivars. The polyamines putrescine, spermidine and spermine showed increased abundance in sensitive cultivars under HNT conditions. Correlations between the content of polyamines and 75 other metabolites indicated metabolic shifts from correlations with sugar-phosphates and 1-kestose under control to correlations with sugars and amino and organic acids under HNT conditions. Increased expression levels of ADC2 and ODC1, genes encoding enzymes catalysing the first committed steps of putrescine biosynthesis, were restricted to sensitive cultivars under HNT. Additionally, transcript levels of eight polyamine biosynthesis genes were correlated with HNT sensitivity. Responses to HNT in the vegetative stage result in distinct differences between differently responding cultivars with a dysregulation of central metabolism and an increase of polyamine biosynthesis restricted to sensitive cultivars under HNT conditions and a pre-adaptation of tolerant cultivars already under control conditions with higher levels of potentially protective compatible solutes.Global climate change combined with asymmetric warming can have detrimental effects on the yield of crop plants such as rice ( Environmental changes have an increasing influence on crop yields all over the world due to the reduced availability of agricultural land and water resources, and especially accelerating global climate change (Oryza sativa L.) and for HNT conditions it was 30\u00b0C/28\u00b0C (day/night). First stress symptoms (chlorosis) were visible after 7 d and fully developed after 23 d of HNT. At this time point a clear separation of all cultivars regarding their HNT sensitivity indicated by chlorosis was possible.Seeds, plant material, cultivation and HNT stress treatment were the same as outlined in After 23 d of HNT treatment (48 DAS), fully expanded leaves were harvested randomized, 2\u22124h after the beginning of the light period and immediately frozen in liquid nitrogen. Two independent experiments were performed for both control and HNT conditions in two growth chambers.A detailed phenotypic and physiological characterization, including measurements of photosynthetic yield, respiration, as well as carbohydrates, is described in Metabolite profiling was performed as in 2 transformed. Hierarchical clustering with Pearson correlation was applied using the analysis, visualization and data-mining software MultiExperiment Viewer . PCA was performed using R (www.r-project.org)Average mass spectral intensities over all replicate samples were calculated for every metabolite in each cultivar. Only metabolites identified in all replicate experiments (two control and two HNT experiments) were used for further analysis. Further, these averages were divided by the median of all averages of each metabolite over all cultivars and logAnalysis of polyamines was performed according to Protein content was measured using the amido black assay based on the method of actin-1 (Os03g50890) gene (Quantitative RT-PCR (qRT-PCR) was performed for nine selected cultivars according to actin-1. Relative expression was calculated as 2\u2013\u0394Ct, with \u0394Ct calculated by subtracting the average Ct values from three technical replicates of the housekeeping gene from the averages of the gene of interest. Fold change was calculated as log2 of the ratio of relative expression of genes under stress conditions to relative expression under control conditions.Data were analysed using the SDS 2.0 software (Applied Biosystems) and normalized based on the expression of the housekeeping gene p>0.01; **, 0.01>p >0.001; ***, p<0.001. Correlation analysis was performed with SigmaPlot 12.3 using Spearman\u2019s rank order correlation.The significance of differences between measured values was analysed using paired or unpaired, two-sided T-tests with SigmaPlot 11.0 . Significance levels are indicated as: *, 0.05>japonica or indica subspecies (Supplementary Table S1) with different HNT tolerance were investigated under HNT (30\u00b0C day/28\u00b0C night) and control conditions (28\u00b0C day/21\u00b0C night). Classification of HNT tolerance was based on ranks of a characteristic leaf chlorosis phenotype (To investigate the impact of high night temperature (HNT) on central metabolism, 12 previously characterized rice cultivars from either the Metabolite profiles were investigated for leaf blades after 23 d (48 DAS) under HNT or after 48 d under control conditions using GC-MS analysis. The vegetative stage was used to investigate symptoms of HNT sensitivity very early during development when the damage by chlorosis is already clearly separating sensitive and tolerant cultivars.Supplementary Table S2). Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were performed on a reduced dataset of 75 metabolites common to all four experiments. Principal components (PC) 1 and 2, which together explained 51.7% of the total variance, clearly separated the metabolite profiles of the three most sensitive cultivars under HNT conditions from all others . Selected representative metabolites of PC1 and PC3 with the 20 highest PC loading values responsible for this separation are shown in Supplementary Table S3.In total, 156 metabolites were identified in two control and two HNT experiments and annotated according to the MPIMP-Golm inventory list (l others . Loadingl others . For PC2l others . These mHCA was performed using Pearson correlation and resulted in five main clusters (CI\u2212V) of metabolites with similar response patterns . In agreSupplementary Fig. S2, Cluster CIII-CV) with M202 again showing the highest number of positive fold changes, followed by DR2 and IR62266-42-6-2.Supplementary Table S4). HNT sensitivity is expressed as a rank determined by leaf chlorosis estimates, with high ranks representing HNT sensitivity were found under HNT conditions. An overlap of six metabolites between the two data sets including erythronic acid, glycerophosphoglycerol, pyroglutamic acid, serine, threonic acid and the unknown metabolite A217004 (not shown) pointed to an equal importance of these metabolites for a more pronounced chlorosis phenotype and reduced FW under HNT. To visualize metabolic pathways affected by HNT conditions, log2 fold changes in the pool sizes of metabolites of glycolysis, the TCA cycle and related pathways, mainly for the biosynthesis of amino acids, are shown in When a correlation analysis was performed between metabolite pool sizes and previously collected FW data, which negatively correlated with HNT .A possible reason for elevated amino acid levels after a stress treatment is increased protein degradation. Total protein content was therefore measured in the same plant material, but no significant changes were detected under HNT compared to control conditions (One striking difference between sensitive and tolerant cultivars after 23 d of HNT was the increase of putrescine (Put), which was also significantly positively correlated with HNT sensitivity. Put is the first compound in the synthesis of spermidine (Spd) and spermine (Spm) and all three polyamines are known to be involved in various stress responses in plants (p=0.0076), Spd and Spm under HNT but not under control conditions.Under control conditions Put levels were highly cultivar dependent with the lowest levels in the sensitive cultivar M202 . In agreSince the content of all three polyamines was significantly correlated with the HNT sensitivity of the cultivars, we wanted to further elucidate the connection of this pathway with the central metabolism. We therefore performed a Spearman\u2019s rank order correlation analysis of polyamine pool sizes with those of all other measured metabolites. Significant correlations differed widely between control and HNT conditions, with six and 19 correlations, respectively . While SInterestingly, the contents of Spm and glutamic acid were correlated under HNT conditions. Glutamic acid is a precursor for polyamine biosynthesis and is derived from the TCA cycle via 2-oxo-glutaric acid . In addiSupplementary Table S5).We monitored the expression of 17 genes encoding polyamine biosynthetic enzymes to investigate whether the differences in polyamine levels in the cultivars were related to differential gene expression. Transcript abundance was measured by qRT-PCR using RNA from the same leaf samples used for polyamine analysis from eight selected cultivars . The selected genes encode enzymes for Put synthesis starting directly from ornithine via ornithine decarboxylase (ODC) or indirectly from arginine via agmatine including arginine decarboxylase (ADC), agmatine iminohydrolase (AIH) and N-carbamoylputrescine amidohydrolase (CPA) . However, ADC2 transcript levels were not significantly correlated with HNT sensitivity (Supplementary Table S7). AIH showed a similar expression pattern but in this case, a significant negative correlation with HNT sensitivity was observed. Similar correlations were also found for CPA1, SPD/SPM1 and SPD/SPM2 . As under control conditions, the most highly expressed genes in all cultivars were SAMDC1 and 2, although expression of both genes was slightly reduced in most cultivars. SAMDC1 expression was positively correlated with HNT sensitivity and the same was true for the expression levels of ADC2, CPA4, ODC1 and SPD/SPM3 between the log2 fold change in expression between control and HNT conditions and HNT sensitivity ranks of the cultivars (Supplementary Table S7). This indicates that the strong down-regulation of polyamine biosynthetic genes in tolerant cultivars were negatively correlated with HNT sensitivity under control conditions (i.e. higher content in tolerant cultivars), while five metabolites showed positive correlations. An induction of genes encoding enzymes involved in the shikimate pathway and related aromatic amino acid metabolism is known as a response to abiotic stress and shikhttp://www.irgcis.irri.org). An additional potentially adaptive reaction of tolerant cultivars under HNT conditions was the reduced expression of almost all genes encoding polyamine biosynthetic genes. In combination with the up-regulation of ADC2, ODC1 and SPD/SPM3 in sensitive cultivars, this resulted in strong correlations of expression levels with the HNT sensitivity rank. The log2 fold change ratios of gene expression of eight out of 17 polyamine biosynthetic genes were positively correlated with HNT sensitivity in agreement with higher polyamine levels in sensitive cultivars. Interestingly, however, the down-regulation of gene expression in tolerant cultivars was not reflected in reduced polyamine levels suggesting that maybe polyamine turn-over rates are important. The ratio of polyamine catabolism and polyamine anabolism has been suggested as the crucial factor in polyamine-mediated stress tolerance of metabolite profiles measured by GC-MS after 48 DAS under control conditions in 12 cultivars of rice.Supplementary Fig. S2. (A) Hierarchical cluster analysis (HCA) with Pearson correlation used for log2 fold changes of 75 metabolites measured by GC-MS after 23 d (48 DAS) of HNT in comparison to control conditions in 12 cultivars of rice. (B) Normalized responses of selected metabolites represent the response patterns of clusters CI\u2212V.Supplementary Fig. S3. Protein content in leaves of 12 rice cultivars after 48 DAS under control and 23 d (48 DAS) under HNT.Supplementary Table S1. Cultivars of Oryza sativa L. used for HNT stress experiments.Supplementary Table S2. Normalized metabolite profiles measured by GC-MS after 48 DAS under control and 23 d (48 DAS) under HNT conditions for all samples.Supplementary Table S3. Top ten most positive and negative loadings of PC1 and PC3 separating indica and japonica cultivars in a PCA of metabolite profiles measured by GC-MS after 48 DAS under control conditions.Supplementary Table S4. Spearman\u2019s correlations for 75 metabolites with HNT sensitivity of genes encoding enzymes involved in polyamine biosynthesis after 48 DAS under control and 23 d (48 DAS) under HNT conditions and the respective log2 fold change in nine cultivars of rice.Supplementary Table S7. Spearman correlation calculated for 17 genes encoding enzymes involved in polyamine biosynthesis with HNT sensitivity [expressed as rank of damage by chlorosis, (2 fold change."} +{"text": "After cardiopulmonary resuscitation (CPR), following an out of hospital cardiac arrest (OHCA) hemodynamic failure is common, due to a combination of heart failure and ischemia reperfusion injury. Comatose post-cardiac arrest patients are treated on the intensive care unit (ICU) with mild therapeutic hypothermia (33\u00b0), nowadays referred to as target temperature management (TTM) for an assumed neuroprotective effect.In previous reports both microcirculatory alterations and impaired vascular reactivity have been described in post cardiac arrest patients treated with mild therapeutic hypothermia. As of now it is unknown whether these alterations are related to the temperature management itself. Aim of the present study was to investigate the potential difference in microcirculatory alterations and vascular reactivity in patients after out of hospital cardiac arrest treated with target temperature management of 33\u00b0C (TTM33) in comparising to patients treated with 36\u00b0C (TTM36).Our study was designed as a a priory substudy of the open label randomized controlled TTM trial in 2 Dutch mixed ICU's. Microvascular flow index (MFI) was assessed by Side Stream Darkfield imaging and vascular reactivity by near infrared spectroscopy. Variables, including systemic haemodynamics were recorded at start study (T1), after 12 hours (T2) and after 24 hours (T3).22 patients were included, 13 in TTM33 and 9 in TTM36. At T1 MFI between groups did not differ significantly . The difference between groups remained insignificant over time. At T1 tissue oxygenation (StO2) was significantly lower in TTM36 in comparison to TTM33 . Over time this difference between groups disappeared. However, vascular reactivity, expressed as the descending and ascending slope of StO2 after a standardized ischemic occlusion test was similar between groupsIn this relatively small sample size study microcirculatory blood flow and vascular reactivity did not change between TTM33 and TTM36Clinicaltrials.gov nr. NCT01850485"} +{"text": "Noninvasive ventilation through a mask is commonly applied in pressure support ventilation (nPSV). Recent studiesshowed that noninvasive neurally adjusted ventilatory assist (nNAVA) improves patient-ventilator interaction and synchrony. More recently we described a new setting for nNAVA (nNAVA15) able to reduce the peak of electrical activity of the diaphragm (EAdipeak) and dyspnea , compared with both nPSV and nNAVA, in patients undergoing NIV through a helmet, by improving the rate pressurization. We therefore designed a physiological study to evaluate and compare the effects of nNAVA15 with nPSV and nNAVA on VASd, EAdipeak, pressurization rate and arterial blood gases (ABGs).2O, fastest rate of pressurization); nNAVA (NAVA level to achieve a comparable EAdipeak as during nPSV); nNAVA15 (NAVA level at 15 cmH2O/jV and the maximum inspiratory airway pressure (Paw) set at the value corresponding to PEEP + inspiratory support during nPSV). Oxygen inspiratory fraction and PEEP remained unmodified throughout the study period. The last minute of each trial was analyzed. Paw-time products of the initial 200 ms from the onset of ventilator pressurization (PTP200), of the initial 300 and 500 ms from the onset of the EAdi swing indexed to the ideal PTP , and of the triggering area (PTPt) were computed. ABGs and VASd were assessed at the end of each trial.Fourteen patients undergoing noninvasive ventilation because of acute respiratory failure underwent three randomized 30-minute trials: nPSV (inspiratory support above positive end- expiratory pressure (PEEP) \u22658 cmHP < 0.001), without affecting ABGs and EAdipeak. nNAVA15 improved PTP300i and PTP500i and 63% %, respectively) compared with nPSV % and 44 %, P < 0.001) and nNAVA % and 46 %, P < 0.001). PTP200 was lower in nNAVA cmH2O*second) with respect to both nPSV cmH2O*second) and nNAVA15 cmH2O*second) (P = 0.001). PTPt was significantly improved by both nNAVA cmH2O*second) and nNAVA15 cmH2O*second) as opposed to nPSV cmH2O*second, P < 0.001).nNAVA15 reduced VASd ), compared with both nPSV ) and nNAVA ) (Compared with nPSV and nNAVA, nNAVA15 through a mask reduces VASd, assuring an optimal pressurization rate and triggering performance, without affecting the breathing pattern, neural drive and ABGs."} +{"text": "New Daily Persistent Headache (NDPH) is a chronic primary headache with uncertain outcome. Comorbities may play important role in choosing the right treatment in order to improve the management of this disorder.To investigate the comorbidity of depression or anxiety of the patients suffering from NDPH.One hundred and forty four (144) headache outpatients (62 men and 82 women) and one hundred and fifty (150) healthy controls (56 men and 94 women) were screened prospectively using a questionnaire that included the Hamilton scales for both Anxiety and Depression.The mean scores of both HAM-A and HAM-D scales were significantly higher in NDPH patients (19\u00b17.3 and 16\u00b16.3 respectively) comparing with the healthy population . No significant difference was detected in the Hamilton scaling scores between NDPH and Chronic Tension Type Headache sufferers.NDPH sufferers showed more depression or anxiety symptoms than healthy controls, as headache sufferers from chronic TTH. Treating properly psychiatric comorbities in NDPH sufferers, or choosing drugs that influence both conditions simultaneously may improve outcome.No conflict of interest."} +{"text": "Combined BRAF and MEK inhibition may delay the onset of resistance compared with BRAF inhibition alone. The safety and tolerability of the BRAF and MEK inhibitors, vemurafenib and cobimetinib were evaluated in a phase 1B trial, BRIM7 (NCT01271803). Modulation of signaling pathways, transcriptional outputs and T-cell dynamics were assessed in tumor samples obtained from BRIM7.+ T-cells in tumor biopsies were assessed by immunohistochemistry. Changes in transcription profiles upon treatment were measured by qRT-PCR using the Fluidigm/Nanostring platforms.Tumor tissues were collected at baseline prior to treatment, on-treatment at cycle 1 day 14 (C1D14) and at disease progression (PD). Modulation of the MAPK and PI3K pathways, cell proliferation, and CD8133 tumor samples have been collected representing 82 unique patients. There are six paired biopsies (baseline and C1D14) from the same patients, and seven evaluable PD samples. In the six paired biopsies (baseline and C1D14), robust inhibition of the MAPK pathway was observed in tumors following vemurafenib + cobimetinib treatment as measured by reduction of pERK levels compared to baseline (mean H-score inhibition 88 +/- 12%). This was observed in samples from BRAF inhibitor-na\u00efve patients (n=2) and also from patients who had progressed while on vemurafenib (n=4). The inhibition of pERK correlated with the reduction of the proliferation marker Ki67 in C1D14 tumor samples (77 +/- 10%), while inhibition of the PI3K pathway marker pS6 showed greater variability (68 +/- 28%). At PD, varying levels of MAPK pathway activity and moderate-to-high expression of Ki67 were seen in tumors. Ongoing analyses of combination treatment effects on the MAPK pathway, immune regulatory gene transcription and CD8+T-cell status will be presented.The combination of vemurafenib + cobimetinib results in MAPK pathway inhibition in both BRAF inhibitor na\u00efve patients and patients who have progressed on vemurafenib. Increased levels of pERK and Ki67 at PD suggest the renewal of proliferation as tumors may have escaped the inhibitory effects of the combination therapy. Effects of the combined treatment on the MAPK pathway and immune gene signatures will also be discussed."} +{"text": "LPA is generated through the action of autotaxin or phospholipases, and degradation begins with lipid phosphate phosphohydrolase (LPP)-dependent removal of the phosphate. While the effects of LPA on ovarian cancer progression are clear, the effects of LPA metabolism within the tumor microenvironment on peritoneal metastasis have not been reported. We examined the contribution of lipid phosphatase activity to ovarian cancer peritoneal metastasis using mice deficient in LPP1 expression. Homozygous deletion of LPP1 (LPP1 KO) results in elevated levels and decreased turnover of LPA in vivo. Within 2 weeks of intraperitoneal injection of syngeneic mouse ovarian cancer cells, we observed enhanced tumor seeding in the LPP1 KO mice compared to wild type. However, tumor growth plateaued in the LPP1 KO mice by 3 weeks while tumors continued to grow in wild type mice. The decreased tumor burden was accompanied by increased apoptosis and no change in proliferation or angiogenesis. Tumor growth was restored and apoptosis reversed with exogenous administration of LPA. Together, these observations demonstrate that the elevated levels of LPA per se in LPP1 KO mice do not inhibit tumor growth. Rather, the data support the notion that either elevated LPA concentration or altered LPA metabolism affects other growth-promoting contributions of the tumor microenvironment.Lysophosphatidic acid (LPA) is a bioactive lipid that enhances ovarian cancer cell proliferation, migration and invasion Lysophsphatidic acid (LPA) is a bioactive lipid that regulates several cellular functions critical for tumorigenesis and metastasis including proliferation, survival, cytoskeletal reorganization, migration, invasion and cytokine production \u20135. The i1 (PLA1) and PLA2 families remove a fatty acid chain from phosphatidic acid to form LPA. Autotaxin (ATX), an extracellular lysophospholipase D also generates LPA following the removal of choline from lysophosphatidylcholine. PLA2 and autotaxin are elevated in ovarian cancer patients [LPA is produced by a variety of cells within the tumor microenvironment including platelets, mesothelial cells, adipocytes, endothelial cells and ovarian cancer cells ,13, and patients ,17, and patients .LPA catabolism is initiated by lipid phosphate phosphohydrolases (LPPs), types 1, 2 and 3, which remove the phosphate to generate monoacylglycerol (MAG). MAG is further cleaved by monoacylglycerol lipase to release the fatty acid chain from glycerol. LPP1 and LPP3 expression is reduced in human ovarian cancers relative to normal ovarian tissue , while fin vitro, the ability of LPA to promote ovarian cancer invasion and growth has been demonstrated in mouse models of peritoneal metastasis; daily injection or implantation of pumps producing high concentrations of LPA increased tumor burden in immune compromised and syngeneic mouse models [In addition to regulating ovarian cancer cell proliferation, survival, migration and invasion e models ,22. Howee models and synge models . Lipid pe models . Here, wID8 cells were generously provided by Dr. K. Roby (University of Kansas) and cult6 ID8ip2Luc cells in 200 \u03bcL PBS. Experiments were performed with groups of 3\u20135 mice per genotype and/or treatment; the total number of mice analyzed (n) is indicated in the figure legend. Mice were observed 2\u20133 times per week by laboratory personnel and monitored for signs of distress and weighed; mice showing signs of distress or losing greater than 15% body weight were euthanized and examined for tumor. LPA was suspended in 0.5% fatty acid-free bovine serum albumin in phosphate-buffered saline (PBS) at a concentration of 100 \u03bcmol/L, and 200 \u03bcL were delivered IP once daily beginning 1 day after tumor initiation and continued for 6 weeks. Daily administration of vehicle served as negative control. Tumor burden was monitored weekly by measuring light emission following IP luciferin administration as an indication of luciferase activity using an IVIS imaging system . Total flux (photons/sec) was determined for the entire abdominal cavity. Upon experimental termination, mice were euthanized and tumor burden evaluated upon necropsy by counting the number of tumor nodules, and weighing the omentum (primary site of tumor implantation) and any additional tumor nodules. Formalin-fixed, paraffin-embedded tissues were sectioned and H&E stained (University of Virginia Research Histology Core) to evaluate microscopic tumor burden, the extent of peritoneal invasion, and mitotic index.All animal experiments were performed following approval from the Institutional Animal Care and Use Committee at the University of Virginia. Six to 8 week-old female wild type (C57BL/6) or LPP1 hypomorphic (LPP1 KO) mice were injected intraperitoneally (IP) with 10One section of tumor containing omentum per mouse was subjected to immunohistochemistry for Ki67 , cleaved caspase 3 , CD31 , or VCAM-1 (University of Virginia Biorepository Tissue Research Facility). Briefly, sections were boiled in low pH or high pH EnVision FLEX Target Retrieval Solution (Dako) and stained with the indicated concentrations of each antibody. Antigen\u2013antibody complex was detected using Envision Dual Link (Dako) followed by incubation with 3,3\u2019-diaminobenzidine tetrahydrochloride (DAB+) chromogen (Dako) and counterstained with hematoxylin. Ki67, and cleaved caspase 3 staining was quantified by summing the number of positively and negatively stained tumor cells in 5 high-powered (400X magnification) fields per section (one section per mouse) to calculate the percent positive cells. The extent of vessel formation was determined by counting the number of CD31 stained vessels and normalizing it to the total tumor area per 200X field . The percentage of mesothelial cells expressing membranous VCAM-1 was scored positive at >50%.Growth factor-reduced matrigel (BD Biosciences) was mixed with ID8ip2Luc cell conditioned media (1:1) or with 1 \u03bcg/ml FGF, 20 \u03bcg/ml VEGF and injeAll data were analyzed using GraphPad Prism 6 software. Luminescence data were analyzed using 2-way analysis of variance (ANOVA) followed by Tukey\u2019s multiple comparisons test. One-way ANOVA followed by Tukey\u2019s multiple comparisons test was used to analyze omentum weights and tissue staining from mice treated with LPA or vehicle. The incidence of invasive tumors or VCAM-1 staining was analyzed using Fisher\u2019s Exact test. The remaining data were analyzed with an unpaired t-test or the Wilcoxan-Mann-Whitney test depending on whether the data were parametric.In vivo imaging of mice following intraperitoneal (IP) injection of ID8ip2Luc cells revealed increased luminescence in LPP1 KO mice compared to wild type within 2 weeks factor, the presence of an inhibitor of growth (inducer of cell death), the inability to recruit auxiliary cells necessary to promote growth, inappropriate cell-cell interactions within the tumor or any combination of these or additional effects. We hypothesized that exogenous administration of high concentrations of a growth factor would override the negative consequences of the growth inhibitory environment. LPA is a well-established stimulator of ovarian cancer cell growth ,4. DailyLPA is an important regulator of ovarian cancer growth and metastasis. In addition to the increased levels found in ascites, which is accompanied by increased ATX activity, the tumors themselves have de novo expression of LPA receptors LPA2 and LPA3) and LPA3,20. Herein vivo [LPP1 KO mice are characterized as having increased plasma LPA levels and reduced LPA turnover . LPA stiin vivo , althougin vivo . InteresWhile LPP1 KO mice have increased tumor seeding in the presence of elevated levels and reduced turnover of LPA, subsequent tumor growth was stunted and accompanied by increased apoptosis, a result that contradicts well-established pro-growth, pro-survival effects of LPA. The mechanisms by which loss of LPP1 in the tumor microenvironment halts tumor growth are unclear. One possibility is that elevated LPA as a result of LPP1 deficiency affects other cellular components of the tumor microenvironment to negatively impact tumor growth. The tumor microenvironment contains a complex mixture of cytokines, growth factors and multiple cell types, including endothelial and immune cells that function together to promote tumor growth . While tThe effects of loss of LPP1 on tumor growth could be due to alterations in the metabolism of other phosphoglycerides. In addition to LPA, LPP1 acts on other phosphoglycerides including phosphatidic acid and sphingosine 1-phosphate (S1P) , which aTogether, the data presented here indicate that the loss of LPP1 creates an environment insufficient to support tumor growth. Exogenous supply of higher concentrations of LPA was able to overcome the inhibitory environment to stimulate tumor growth and diminish apoptosis demonstrating that LPA stimulates tumor cell growth regardless of any potential negative effects created by the loss of LPP1 expression. Further investigation of the mechanisms responsible for reduced tumor growth in the absence of LPP1 would provide additional important information regarding the effects of phosphoglycerides and their metabolism on the tumor microenvironment as well as components within the microenvironment that might be important in promoting ovarian cancer progression.S1 Fig(A) Omentums were removed and weighed 2 weeks after tumor initiation from wild type and LPP1 KO mice. Each point indicates a single individual, and the data represent the mean \u00b1 std err. (B) H&E stained sections of omentum were evaluated for microscopic tumors and the extent of invasion. Arrows indicate the mesothelium; arrowheads indicate tumor. Non-invasive tumors were characterized as having a smooth interface between the tumor and underlying tissue. Invasive tumors were scored as those spidering through and taking over the underlying tissue, in this case, omental fat. (C) Omentums from wild type (top panel) and LPP1 KO (bottom panel) mice were obtained 2 weeks after tumor initiation and stained for VCAM-1 expression using IHC (see IHC see . Represe(TIF)Click here for additional data file.S2 Fig(A) or FGF/VEGF (B) are shown.Representative images of CD31 positive vessels (indicated by arrows) in matrigel plugs containing conditioned media from ID8ip2Luc cells (TIF)Click here for additional data file."} +{"text": "Antiphospholipid antibodies (aPL) play the central pathogenic role in antiphospholipid syndrome (APS) characterized by arterial and venous thrombosis, recurrent fetal loss and persistent circulating aPL. The diagnostic criteria of definite APS are not entirely applicable in pediatric population. An international multicentric project named Ped-APS registry included 121 patients with APS onset before 18th birthday. Almost half of them (49.5%) had an associated autoimmune disease. In this study a large percentage of children with aPL-related thrombotic event had at the time of the initial thrombotic event associated nonthrombotic clinical manifestations including: hematological manifestations (38%), dermatological manifestations (18%) and nonthrombotic neurological manifestations (16%).2-glycoprotein I antibodies (anti-\u00df2GPI) and lupus anticoagulant (LA), among children with positive laboratory results when tested for the presence of aPL.To evaluate the spectrum of thrombotic and non-thrombotic clinical manifestations associated with aPL, namely anticardiolipin antibodies (aCL), anti-\u00df2GPI and/or positive for LA at least once in the period from January 1997 to July 2012. Clinical manifestations of these patients were then evaluated, specifically thrombosis, nonthrombotic neurological and hematological manifestations, skin disorders and cardiac valve disease.Pediatric patients in a tertiary care hospital were according to the clinical judgment of the treating physician tested for the presence of aPL and the laboratory results were saved in a computer database. In this single-center bidirectional study we included 159 consecutive patients from the database that tested medium or highly positive for aCL and/or positive for anti-\u00dfOf all 159 patients 55 had an underlying systemic disease and 8 had other autoimmune disease. Sixty-six out of 159 (42%) patients presented with one aPL-related clinical manifestation, 18 (11%) with two and 5 (3%) patients presented with three or more aPL related clinical manifestations. Of the aPL related clinical manifestations thromboses occurred in 25 patients , nonthrombotic neurological disorders were present in 25 patients , hematological disorders in 48 , skin disorders in 19 and a cardiac valve disease in 5.In an unselected group of children with positive aPL nonthrombotic clinical manifestations were more frequent than thrombotic events. The most common clinical manifestations associated with aPL in children are thrombocytopenia, followed by venous thrombosis, seizures and Raynaud's phenomenon.None declared."} +{"text": "Bicuspid aortic valve (BAV)-related aortopathy is characterized by histologic abnormalities that result in aortic wall stiffening. Aortic stiffness has been shown to be an independent predictor of cardiovascular mortality in many settings. We sought to determine the range of aortic stiffness seen in unselected patients with BAV, and investigate associations between stiffness and various standard clinical and imaging parameters.BAV (n=65) and normal patients (n=10) were studied at one time point with axial conventional phase-contrast MRI through the ascending aorta. Aortic stiffness was estimated by measuring pulse wave velocity (PWV) using the flow-area (QA) method. Repeat PWV measurements were made by two independent reviewers, and average values were used for analysis. Associations between imaging and clinical/demographic parameters were investigated with Pearson's correlations. Multiple linear regression models were performed to identify independent predictors of PWV.Inter-rater reproducibility of PWV measurements was high (ICC= 0.97). There was no significant difference in age between BAV and normal patients . There was an overall trend toward higher PWV in patients with BAV compared to normal patients with a considerably higher standard deviation in BAV patients (SD of 5.88 versus 0.92 m/s).-Swedish Research Council (PD).-NIH T32 Training Grant 5T32EB001631-10 (NSB)."} +{"text": "To assess the relationship between the presence of myocardial interstitial fibrosis as reflected by the increase in native T1 values and alterations in left ventricular (LV) diastolic function evaluated by phase contrast cardiac magnetic resonance (PC-CMR), in subjects with severe aortic valve stenosis (AVS).We studied 20 subjects (71\u00b110 years) with severe AVS including 19 with a preserved ejection fraction. All patients underwent transthoracic echocardiogram (TTE) and cardiac magnetic resonance (CMR) exams. CMR included conventional LV systolic function and delayed enhancement evaluations as well as a native T1 mapping acquisition using the modified Look-Locker inversion recovery sequence and velocity encoding data of the transmitral inflow for the evaluation of LV diastolic function. These latter CMR data were analyzed using custom software resulting in segmental T1 values and diastolic parameters such as transmitral peak velocities , peak flow rates , filling volume (FV), and myocardial peak velocities.For all patients, TTE revealed the presence of severe AVS according to ESC criteria . When compared to CMR data of 34 elderly controls (60\u00b18 years) despite the preserved ejection fraction , diastolic parameters indicated an impaired LV relaxation in patients with severe AVS. Importantly, while dense fibrosis volume quantified from delayed enhancement images was not related to diastolic function parameters, a significant relation was found between native myocardial T1 values and parameters of LV filling such as: the ratio between the peak filling rate and the peak atrial rate EfMR/AfMR ; the ratio between the peak atrial rate and the filling volume Af/FVMR ; and the peak atrial rate Af .Interstitial myocardial fibrosis assessed non-invasively by native T1 is related to the severity of diastolic dysfunction in subjects with severe AVS.This study was funded by Assistance Publique des H\u00f4pitaux de Paris."} +{"text": "In HIV+ patients exhibiting multidrug resistance (MDR), NRTIs often have little activity, increased toxicity, drug interactions and add unnecessary treatment costs. The 48 week VERITAS study demonstrated that these patients can have a safe and effective simplification of salvage regimen by removing inactive NRTIs as determined by genotypic data. Virological, immunological, clinical and financial outcomes were evaluated at an additional 96 weeks of follow-up.MDR patients with an undetectable viral load (VL) on a stable regimen containing at least four ARVs (including one inactive NRTI) were enrolled in an open-label, prospective simplification trial, where one inactive NRTI was removed at baseline (BL). A second NRTI could be removed at week 24 if the regimen contained at least five ARVs at enrolment.31 male patients participated. The mean length of treatment was 14 years, with a median CD4 count of 525. The BL regimen consisted of 4 ARVs in 22 patients (71%) and 5 ARVs in 9 patients (29%). 3TC or FTC was removed in 29 patients (94%), and either AZT or TDF was interrupted in 2 others. Four patients had a second NRTI stopped. One patient was removed at W26 as an active NRTI was removed for creatinine elevation. 30 well-controlled patients continued follow-up after W48. At W144, six patients had additional changes in their ARV regimen. Half were due to toxicity while the other half were the result of treatment simplification. None of the patients exhibited virologic failure at the time of treatment change and maintained undetectable VLs throughout the entire follow-up. These six patients had a mean gain of 79 CD4 (p=0.17) compared to baseline. 22 of the 24 patients (92%) with no changes in ARV therapy after W48 had undetectable VLs. The other two had confirmed virologic failure, one with genotypic resistance. All 24 had elevated CD4 counts . No deaths or serious adverse events were observed. One or two ARV removals translated to a mean annual saving of $3319 CDN (11%) and $8630 (24%) respectively.Final results indicate that removing one or two inactive NRTIs from a regimen in patients taking four or more ARVs with controlled VL appears to be safe, maintains virological suppression through 144 weeks and significantly reduces treatment costs."} +{"text": "The experiment was conducted to investigate potential causes of grain sterility in widely cultivated rice variety in Malaysia, MR219 and its two mutant lines (RM311 and RM109) by examining the source-sink relations. RM311 produced increased dry matter yield both at heading and maturity and also showed higher grain yield with greater proportion of grain sterility than the other two genotypes (RM109 and MR219) resulting in the lowest harvest index (49.68%). In contrast, harvest index was greater in RM109 (53.34%) and MR219 (52.76%) with less grain sterility percentage than MR311 indicating that dry matter partitioning to economic yield was better in RM109 and MR219 than in MR311. Results indicated that dry matter allocation per spikelet from heading to maturity was important for reducing grain sterility in rice. The greater above-ground crop dry matter per spikelet was observed in RM109 and MR219 as compared to high dry matter producing genotype; RM311 implies that poor grain filling may not have resulted from dry matter production or source limitation. These findings suggest that grain sterility or poor grain filling in rice is the result of poor translocation and partitioning of assimilates into grains (sink) rather than of limited biomass production or source limitation. Oryza sativa L.) is one of the most important food crops in the world. Rice is consumed by more than 50% of the world's population which provides 45\u201360% of the dietary calories \u00f7 NSC in stems at heading \u00d7 100 . TransfeThe collected data were analyzed statistically following the analysis of variance (ANOVA) technique and the mean differences among the treatments were compared by LSD using the statistical computer package program, MSTAT-C.\u22121 (\u22121) due to higher number of tillers hill\u22121, spikelets panicle\u22121, and larger grain size. In contrast, the lowest grain yield was recorded in RM109 (21.14\u2009g hill\u22121) for fewer number of spikelets panicle\u22121. However, filled spikelet (grain plumpness) percentage of RM311 was the lowest (68.9%) which was significantly less than the other two genotypes, RM109 and MR219. The highest filled grain percentage was recorded in MR219 (79.2%) and this genotype also showed the lowest partially filled (12.4%) and unfilled grains (8.4%) followed by RM109 . The results indicate that a greater number of spikelets per panicle did not show maximum efficient yield potential because of their low filled spikelet percentage. Results further revealed that spikelet sterility was positively correlated with number of spikelets panicle\u22121 and grain size (The effect of genotypes on grain yield and yield attributes in rice was significant except for the number of tillers hill\u22121 . The higain size . Yamamoty filled .4% and u\u22121) and heading to maturity (20.11\u2009g\u2009hill\u22121) followed by MR219 . The greater biomass production both at heading and heading to maturity of RM311 associated with higher leaf area index (LAI) and photosynthesis (Pn) rate at heading with the highest unfilled grains (12.1%), while RM109 produced the lowest TDM at heading (25.12\u2009g\u2009hill\u22121) with less unfilled grains (8.8%). Results further revealed that the proportion of dry matter production from heading to maturity was greater in MR109 (37.8%) and MR219 (38.8%) than in RM311 (33.4%) suggesting that the increase in dry matter accumulation after heading would be greatly beneficial to enhance grain filling process because 80\u2013100% of the yield comes from assimilates which are produced during grain filling period [There was a significant variation in above-ground dry matter accumulation and harvest index (HI) in rice genotypes . The geng period . This re\u22121) and also showed the lowest grain sterility . The greater above-ground crop dry matter per spikelet was found in RM109 and RM219 with lower LAI as compared to high TDM producing genotype (RM311) which implies that poor grain filling may not have resulted from dry matter production or source limitation. This result indicates that grain sterility is mainly due to insufficient assimilate supply to grain. The percentage of remobilized carbon (C) reserve and transfer ratio of total assimilate (TRA) to grains during grain filling period were lower in RM311 than the other two genotypes, RM109 and MR219 . The low remobilization of assimilates of RM311 resulted in a large amount of nonstructural carbohydrate (NSC) left in stems at maturity (106.6\u2009mg\u2009g\u22121\u2009dw) causing greater grain sterility , thereby poor harvest index. These results indicate that grain sterility is mainly due to low translocation rate from source to sink since little C reserve was remobilized and much of the NSC remained in the stems at maturity.Considering the dry matter allocation per spikelet, results showed that dry matter accumulation from heading to maturity was higher in MR219 and RM109 than in RM311 , though The pattern of photosynthesis (Pn) rate and dry weight of stem during the grain growth period were different among the genotypes Figures and 2. PBased on source-sink relationships, rice varieties can be categorized as sink-limiting, source-limiting, or intermediate types . Most inEfficient and higher dry matter allocation per spikelet is more important than increasing the total dry matter production to reduce grain sterility in rice. Grain sterility is mainly due to low translocation rate from source to sink rather than limited biomass production or source limitation. Therefore, it is possible to improve grain filling of indica rice by selecting suitable parents."} +{"text": "Selecting probable idiopathic normal pressure hydrocephalus (INPH) patients for shunt insertion presents a challenge because of coexisting comorbidities and other conditions that could mimic NPH. The characteristic appearance of DESH on brain imaging has been shown to have a high positive predictive value in identifying shunt responsive INPH patients . However, the negative predictive value of this radiological sign was not clearly demonstrated.A single centre retrospective study of probable INPH patients, who underwent ventriculoperitoneal shunt insertion. Case notes were reviewed for clinical presentation, prognostic tests used and postoperative improvement. Shunt responsive INPH patients were identified as those having objective improvement in their walking speed, neuropsychological assessment as well as subjective improvements in gait, continence and memory, one year post operatively. Preoperative images were reviewed for DESH sign (2 independent reviewers). Negative and Positive Predictive Values (NPV and PPV) of DESH sign were determined post analysis.A total of 103 probable INPH patients were included (31 were DESH positive (30%) and 72 were DESH negative (70%)). A total of 78 patients showed measurable improvement 1 year post shunt insertion (76%); 24 (31%) of these patients were DESH positive and 54 (69%) were DESH negative (P = <0.001). Therefore, the DESH sign had a PPV of 77% and a NPV of 25%.In our data, the presence of DESH sign demonstrates a high positive predictive value of 77%, in agreement with SINPHONI trial data. However, it has shown a low negative predictive value. We conclude that probable NPH patients should not be excluded from having shunt insertion based on the presence of DESH sign alone.DESH: Disproportionately Enlarged Subarachnoid Space Hydrocephalus; NPH: Normal Pressure Hydrocephalus."} +{"text": "We found loss of tumoral Eya2 expression in 63% of pancreatic cancers (120/189 cases). Silencing of EYA2 expression in pancreatic cancer cell lines correlated with promoter methylation and histone deacetylation and was reversible with DNA methyltransferase and HDAC inhibitors. EYA2 knockdown in pancreatic cancer cell lines increased cell proliferation. Compared to parental pancreatic cancer cells, pancreatic cancers stably-expressing EYA2 grew more slowly and had fewer metastases in orthotopic models. The transcriptional changes after stable expression of EYA2 in pancreatic cancer cells included induction of genes in the TGFbeta pathway. Epigenetic silencing of EYA2 is a common event in pancreatic cancers and stable expression EYA2 limits the growth and metastases of pancreatic adenocarcinoma.To identify potentially important genes dysregulated in pancreatic cancer, we analyzed genome-wide transcriptional analysis of pancreatic cancers and normal pancreatic duct samples and identified the transcriptional coactivator, EYA2 (Drosophila Eyes Absent Homologue-2) as silenced in the majority of pancreatic cancers. We investigated the role of epigenetic mechanisms of EYA2 gene silencing in pancreatic cancers, performed KRAS, CDKN2A, TP53 and SMAD4 were injected subcutaneously into male CD1 nu/nu athymic mice . One week after the tumor cell injection, subcutaneous tumor volumes (V) were measured weekly with digital calipers (Fisher Scientific) and calculated using the formula V = 1/2(ab2), where a is the biggest and b is the smallest orthogonal tumor diameter. After 3 weeks, the subcutaneous tumors from Panc2.5 stable transfectants and controls were harvested and cut into cubes of approximately 1 mm3 and orthotopic xenografts by surgical implantation were performed as described previously . We . We 43].The presence of transcription factor binding sites within the promoters (from 2000 bp upstream the transcription start site) of potential target genes was assessed using the MAPPER Search Engine . A custoTotal cellular extracts from Panc2.5- and Panc3.014-EYA2-expressing and control clones were prepared using standard procedures and immuStatistical analysis of gene expression array data was completed with Partek Genomic Suite 6.4 software. Raw Affymetrix intensity measurements of all probe sets were background corrected and normalized by the Robust Multichip Average method. Gene expression intensities were summarized by the one-step Tukey's biweight method. Survival rates were calculated by the Kaplan-Meier method, and statistical significance was examined by the log-rank test and the Cox proportional hazards regression model. One-way ANOVA analysis was performed to identify significant expression changes between EYA2-overexpressing and control pancreatic cancer cells and between control pancreatic cancer cells and the parental, non-transfected cell line. P-values of less than 0.05 were considered statistically significant. Statistical analyses were performed using SPSS version 11 ."} +{"text": "We have provided evidence in former studies that cytokines measured in blood correlate negatively with lung function in deltaF508 homozygous patients. GAP junction proteins might be of importance for the influx of blood cells into the lung. Our aim was to assess the relationship between connexin genotypes and cytokines in induced sputum and serum, and lung disease.1(%) 77) were examined. Sequence analysis was performed for genes encoding GAP junction protein alpha 1 (GJA1/connexin 43) and gap junction protein alpha 4 (GJA4/connexin 37). Cytokines were assessed in serum and induced sputum (IS) by chemiluminescence as well as leukocyte counts.36 patients homozygous for deltaF508 in one patient, four common SNPs were detected in GJA4. Two were synonymous changes, but the third variant (rs41266431) predicts an amino acid substitution as well as the fourth SNP . For rs41266431 patients with homozygosity for the G variant had higher IL-8 levels in serum as well as leukocytes in sputum (median: 2050/421 /\u00b5l p=0.041) than those showing heterozygosity (G/A). In individuals > 30 years lung function was worse.DNA analysis was performed in 35 patients. Whereas GJA4 a potential disease modifying gene.SNP rs41266431 seems a promising candidate for further investigations, suggesting"} +{"text": "The number of complicated oncologic patients is increasing accordingly to the high overall prevalence of cancer and the improved survival rate due to advances in cancer treatments. Progression disease and treatment-related complications are the most common causes. In this presentation, we review the pathophysiology of selected oncologic emergencies in which radiologists play a critical role in timely diagnosis and thus have a significant impact on patients\u2019 care.We selected patients who attended the emergency department of our hospital between 2010 to 2014 with different acute symptoms.Of the oncological emergencies presented we found most commonly: compression of the spinal cord (4.5%), carcinomatous meningitis (13.6%); obstruction of the central airway (9%), oesophageal fistula (9%), pulmonary embolism (9%), superior vena cava syndrome (4.5%), pelvic abscess (18%), intra-abdominal haemorrhage (9.5%) and intestinal intussusception (13.6%).Oncologic emergencies represent important causes of death in cancer patients. Radiologists should be familiar with imaging findings of acute conditions to optimize patient care. Recognition of key imaging findings allows prompt diagnosis and facilitates treatment, reducing morbidity and mortality with consequent better outcome."} +{"text": "The development of stable tissue-engineered autologous bone grafts in the field of regenerative medicine is still a challenge. Perfusion bioreactors not only provide continuous nutrition supply and waste removal, but are also suitable for the controlled application of mechanical forces like fluid shear stress. Mechanical loading is known to cause mechanotransductive effects like the induction of differentiation, resulting in enhanced deposition of extracellular matrix .In our study, we determined the optimal flow rate for the osteogenic differentiation of human adipose-derived mesenchymal stem cells (MSC) by applying fluid shear stress that mimics the physiological environment normally experienced by bone progenitor cells in vivo. For this, we first analyzed the porosity of cell substrates with nanofocus-computed tomography as well as their specific permeability at different flow rates. To investigate the effect of controlled application of physiologic fluid shear stress a flow rate of 0.3 ml/min was used to cultivate MSC in a self-developed perfusion bioreactor. Cells were seeded on a three-dimensional macro-porous zirconium dioxide ceramic scaffold (0.3\u00b7106 cells/scaffold) and cultivated in standard growth medium (GM)or osteogenic differentiation medium (ODM) under normoxic (21% O2) or hypoxic (5% O2) conditions for a period of 21 days. After cultivation cell viability was examined using MTT assay. Furthermore DAPI staining was used to evaluate cell distribution. Glucose consumption and lactate production were monitored and histological stainings were used to evaluate osteogenic differentiation.Flow rates between 0.3 - 5 ml/min result in fluid shear stress between 0.01 - 2.5 Pa which is in the range of physiologic shear stress bone progenitor cells are subjected to in vivo (0.3 - 3 Pa)[Physiologic fluid shear stress together with physiologic oxygen conditions (5% O2) lead to higher cell viability. Furthermore the glucose metabolism is elevated under perfusion due to enhanced mass transfer. The application of fluid shear stress results in a stronger differentiation regarding matrix deposition even in standard growth medium without any osteoinductive supplements. These results underline the positive effects of dynamic cultivation and physiologic oxygen concentration which together mimic in vivo conditions. Consequently other factors like medium composition should be adjusted to be more physiologic and taken into consideration during a tissue engineering process to ensure a physiologic tissue maturation.Part of this work was performed with financial support of the FFG BRIDGE project \"3D Tissue\"."} +{"text": "Triticum turgidum L. ssp. durum). Water stress during early reproductive stages can result in significant yield loss in durum wheat and this study describes genotypic differences in the miRNAome between water deficit tolerant and sensitive durum genotypes. Small RNA libraries were constructed from flag leaf and developing head tissues of four durum genotypes, with or without water stress to identify differentially abundant miRNAs. Illumina sequencing detected 110 conserved miRNAs and 159 novel candidate miRNA hairpins with 66 conserved miRNAs and five novel miRNA hairpins differentially abundant under water deficit stress. Ten miRNAs were validated through qPCR. Several conserved and novel miRNAs showed unambiguous inverted regulatory profiles between the durum genotypes. Several miRNAs also showed differential abundance between two tissue types regardless of treatment. Predicted mRNA targets (130) of four novel durum miRNAs were characterised using Gene Ontology (GO) which revealed functions common to stress responses and plant development. Negative correlation was observed between several target genes and the corresponding miRNA under water stress. For the first time, we present a comprehensive study of the durum miRNAome under water deficit stress. The identification of differentially abundant miRNAs provides molecular evidence that miRNAs are potential determinants of water stress tolerance in durum wheat. GO analysis of predicted targets contributes to the understanding of genotypic physiological responses leading to stress tolerance capacity. Further functional analysis of specific stress responsive miRNAs and their interaction with targets is ongoing and will assist in developing future durum wheat varieties with enhanced water deficit stress tolerance.MicroRNAs (miRNAs) are small non-coding RNAs that play critical roles in plant development and abiotic stress responses. The miRNA transcriptome (miRNAome) under water deficit stress has been investigated in many plant species, but is poorly characterised in durum wheat ( Triticum turgidum L. ssp. durum) is the only tetraploid wheat species grown commercially throughout the world. Water deficit stress is one of the main abiotic factors that cause durum yield loss in Mediterranean environments. Water deficit stress in early reproductive stages has been shown to adversely affect grain yield and biomass through reduced grain number in durum as the reference gene.In order to evaluate the expression of miRNA candidates, poly-A tailing combined with qPCR was performed for a select group of seven conserved and three novel stress responsive miRNAs with the 96 durum total RNA samples which were used for sRNA library construction. For each sample, 1 \u03bcg of total RNA was poly-A tailed and reverse-transcribed with the NCode VILO miRNA cDNA synthesis kit following the manufacturer\u2019s instructions. The final cDNA product was diluted to 100 \u03bcL. qPCR was performed using the ViiA\u2122 7 Real-Time PCR system . In each 10 \u03bcL qPCR reaction , 1 \u03bcL diluted cDNA template and primers (3 pmol of each forward and reverse) were mixed with SYBRequences . The revhttp://plantgrn.noble.org/psRNATarget/) with the following parameters: prediction score cut-off value = 3.0, length for complementarity scoring = 20, and target accessibility = 25. Mature novel miRNA sequences were used as queries and the wheat DFCI gene index (TAGI) version 12 was used as the reference genome dataset [http://www.blast2go.com) [\u0394\u0394 CT) method with GAPDH as the reference gene [GenBank: AF251217]. Target primers were designed to include the predicted miRNA/mRNA binding region in the amplified product ensuring the quantification of uncleaved targets, in order to examine the correlation of miRNA and regulated targets. Target transcript sequences, primer locations and primer sequences are listed in The putative mRNA targets of stress responsive novel miRNAs were identified using psRNA Target Server ( dataset . All the2go.com) ,78. BLASS1 Fighttp://www.ncbi.nlm.nih.gov/geo/), and are accessible under the accession number GSE69339.Only one representative library is shown. All sequencing reads were submitted to the NCBI GEO database ((EPS)Click here for additional data file.S2 FigMature miRNAs are highlighted in blue while miRNA* are highlighted yellow. The secondary structures of the novel durum wheat miRNA hairpins (A) Ttu-pre-miR007, (B) Ttu-pre-miR008, (C) Ttu-pre-miR038, (D) Ttu-pre-miR109, and (E) Ttu-pre-miR119 are shown.(TIF)Click here for additional data file.S3 FigIdentified target gene transcripts are searched in the species-specific entries registered in the GO database. Species distribution is based on the number of BLAST hits aligned in each species.(TIF)Click here for additional data file.S1 TableData is shown for 96 libraries presented in 16 different biological library pools (four genotypes \u00d7 two tissue types \u00d7 two treatments). CG = Control group; WG = Water deficit stress group; FL = Flag leaf libraries; H = Head libraries.(XLS)Click here for additional data file.S2 TableCG = Control group; WG = Water deficit stress group; FL = Flag leaf libraries; H = Head libraries.(XLS)Click here for additional data file.S3 TableCG = Control group; WG = Water deficit stress group; FL = Flag leaf libraries; H = Head libraries. The Hairpin Alignment Identifier is derived from the genome location information of the hairpin sequence in the IWGSC CSS , as well as the alignment position and the length of the reads used to identify putative microRNA hairpins. The Hairpin Alignment Identifiers take the following form as an example: 1AL_3896362:3010\u20133120,21. 1AL_(XLS)Click here for additional data file.S4 TableArabidopsis thaliana; Bdi = Brachypodium distachyon; Gma = Glycine max; Hvu = Hordeum vulgare; Osa = Oryza sativa; Sbi = Sorghum bicolor; Tae = Triticum aestivum; Zma = Zea mays.Fold changes have been determined by comparing the RPM between the control treatment libraries and the water deficit stress treatment libraries in the flag leaf and the developing head of four durum wheat genotypes. Fold changes are shown when greater than 1.5 fold. Green values indicate that miRNA reads were more abundant in the water deficit treatment libraries. Red values indicate that the miRNA reads were more abundant in the control treatment libraries. CG = Control group; WG = Water deficit stress group; FL = Flag leaf libraries; H = Head libraries; Be = EGA Bellaroi; Ta = Tamaroi; Tj = Tjilkuri; Ya = Yawa; Ath = (XLS)Click here for additional data file.S5 TableArabidopsis thaliana; Bdi = Brachypodium distachyon; Gma = Glycine max; Osa = Oryza sativa; Zma = Zea mays.Fold changes have been determined by comparing the RPM in Illumina sequencing or comparing relative expression ratio in qPCR between the control treatment and the water deficit stress treatment in different tissues of four durum wheat genotypes. Bold fold change value indicates that the miRNA was more abundant in the water deficit stress treatment libraries whereas unbolded fold change indicates that the miRNA was more abundant in the control treatment libraries. FL = Flag leaf libraries; H = Head libraries; CG = Control group; WG = Water deficit stress group; Be = EGA Bellaroi; Ta = Tamaroi; Tj = Tjilkuri; Ya = Yawa; Ath = (XLS)Click here for additional data file.S6 TableDefinitions: Column E (Expectation)\u2013The expectation scoring of the complementarity between miRNAs and their targets. The maximum expectation threshold score was set at 3.0. Column F (Target Accessibility (UPE))\u2013The maximum energy required to open (unpair) the secondary structure around the target site on the target mRNA. Column O (Multiplicity)\u2013Multiplicity of the target site representing the number of target sites within a specific target transcript.(XLS)Click here for additional data file.S7 TableDefinitions: Column A (Level)\u2013The GO level represents the position of a GO term in the GO hierarchy. The level of a GO term is the number of GO terms between that term and the Root Term of the Ontology. Column E (Node score)\u2013The node score is the sum of sequences directly or indirectly associated to a given GO term weighted by the distance of this term to the term of its direct annotation, i.e. the GO term the sequence is originally annotated to. This confluence score takes into account the number of sequences converging at one GO term and at the same time penalises by the distance to the term where each sequence was actually annotated. Column F (%Seq)\u2013The percentage of sequences annotated with a particular GO term among all the sequences annotated within the same GO level. Column G (#Seq)\u2013The number of target sequences annotated with that particular GO term.(XLS)Click here for additional data file.S8 TableGreen values indicate that the targets were up-regulated under water deficit stress, while red values indicate that the targets were down-regulated under water deficit stress. Bold fold change values indicate negative correlation with Ttu-miR008. FL = Flag leaf libraries; H = Head libraries.(XLS)Click here for additional data file.S9 TableEach forward primer was designed based on the full sequence of the mature miRNA.(XLS)Click here for additional data file.S10 Table(XLS)Click here for additional data file."} +{"text": "In Sebacinaceae, diversification of species with ectomycorrhizal lifestyle presumably started during the Paleocene. Lineage radiations of the core group of ericoid and cavendishioid mycorrhizal Sebacinales started probably in the Eocene, coinciding with diversification events of their hosts. The diversification of Sebacinales with jungermannioid interactions started during the Oligocene, and occurred much later than the diversification of their hosts. Sebacinales communities associated either with ectomycorrhizal plants, achlorophyllous orchids, ericoid and cavendishioid Ericaceae or liverworts were phylogenetically clustered and globally distributed. Major Sebacinales lineage diversifications started after the continents had drifted apart. We also briefly discuss dispersal patterns of extant Sebacinales.Patterns of geographic distribution and composition of fungal communities are still poorly understood. Widespread occurrence in terrestrial ecosystems and the unique richness of interactions of Sebacinales with plants make them a target group to study evolutionary events in the light of nutritional lifestyle. We inferred diversity patterns, phylogenetic structures and divergence times of Sebacinales with respect to their nutritional lifestyles by integrating data from fossil-calibrated phylogenetic analyses. Relaxed molecular clock analyses indicated that Sebacinales originated late Permian within Basidiomycota, and their split into Sebacinaceae and Serendipitaceae Arbutus, Arctostaphylos and Pyrola) mya and that Sebacinales originated around 223 [159\u2013309] mya. Scenario 2 estimated the origin of Basidiomycota at around 521 [452\u2013766] mya and 252 [166\u2013391] mya for Sebacinales . In geneet al. search criteria. The following criteria were used to filter the ITS sequence dataset: a) sequences with a length < 500 bp, b) aligned sequences with less than 60% of longest sequence, c) sequences generated from soil, d) fruit bodies, hyphae or mycelia with unknown ecology, e) without ecological information and f) sequences incorrectly labelled as Sebacinales.(XLS)Click here for additional data file.S4 Table(XLS)Click here for additional data file.S1 Data(FASTA)Click here for additional data file.S2 Datarpb1 and atp6 sequences with a length of 4,438bp.The alignment comprises 18S, 28S, (NEX)Click here for additional data file.S3 DataIncluding a table of all Basidiomycota fossils with species name (if available), times, epochs, citations and calibration usages.(DOC)Click here for additional data file.S4 Data(TXT)Click here for additional data file.S5 Data(XML)Click here for additional data file.S1 FigBootstrap values \u2265 70% are given.(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file."} +{"text": "Cajanus cajan) despite the fact that genome sequence has become available for the crop. To identify the stable housekeeping genes in pigeonpea for expression analysis under drought stress conditions, the relative expression variations of 10 commonly used housekeeping genes were studied on root, stem and leaves tissues of Asha (ICPL 87119). Three statistical algorithms geNorm, NormFinder and BestKeeper were used to define the stability of candidate genes. geNorm analysis identified IF4\u03b1 and TUB6 as the most stable housekeeping genes however, NormFinder analysis determined IF4\u03b1 and HSP90 as the most stable housekeeping genes under drought stress conditions. Subsequently validation of the identified candidate genes was undertaken in qRT-PCR based gene expression analysis of uspA gene which plays an important role for drought stress conditions in pigeonpea. The relative quantification of the uspA gene varied according to the internal controls (stable and least stable genes), thus highlighting the importance of the choice of as well as validation of internal controls in such experiments. The identified stable and validated housekeeping genes will facilitate gene expression studies in pigeonpea especially under drought stress conditions.Gene expression analysis using quantitative real-time PCR (qRT-PCR) is a very sensitive technique and its sensitivity depends on the stable performance of reference gene(s) used in the study. A number of housekeeping genes have been used in various expression studies in many crops however, their expression were found to be inconsistent under different stress conditions. As a result, species specific housekeeping genes have been recommended for different expression studies in several crop species. However, such specific housekeeping genes have not been reported in the case of pigeonpea ( Quantitative real-time PCR (qRT-PCR) is one of the most precise, sensible and widely applied techniques to investigate the candidate genes expression , 2. GeneACT), glyceral-dehyde-3-phosphate dehydrogenase (GAPDH), 18S ribosomal RNA (18SrRNA), 25S ribosomal RNA (25SrRNA), polyubiquitin (UBQ), ubiquitin conjugating enzyme (UBC), elongation factor 1-A (EF1A) and tubulin (TUB) etc. have been used as reference genes in different expression profiling studies in many plant species \u00d7 100%, where S represents the slope of the linear regression line.The X-axis represents the log10 cDNA dilution series, and the Y-axis represents the cycle threshold (Ct). The reaction efficiency (E) is given by [10(TIF)Click here for additional data file.S3 FigDissociation curves for ten housekeeping genes with single peak obtained from two technical replicates of 12 different cDNA pools. X-axis represents temperature (\u00b0C) and Y axis represents Derivative reporter (-Rn).(TIF)Click here for additional data file.S4 FigThis figure shows Ct distribution of each candidate reference gene among the 12 samples calculated through geNorm algorithm.(TIF)Click here for additional data file.S5 FigThis figure shows Ct distribution of each candidate reference gene among the 12 samples calculated through NormFinder algorithm.(TIF)Click here for additional data file.S6 FigThis figure shows a heat map of candidate genes plotted based on Ct mean values. Clustering of genes was based upon the Ct mean values of individual candidate genes across tissues. The detailed description of samples is provided in (TIF)Click here for additional data file.S7 FigThis figure shows a heat map of candidate genes plotted based on Ct mean values. Clustering of genes was based upon the Ct mean values of individual candidate genes across tissues. The detailed description of samples is provided in (TIF)Click here for additional data file.S1 TableThis table shows list of stable housekeeping genes identified under different biotic and abiotic stress conditions in different crops.(DOC)Click here for additional data file.S2 TableThis table shows descriptive statistics of all 10 candidate housekeeping genes used in the study for drought stress conditions using BestKeeper algorithm.(DOCX)Click here for additional data file."} +{"text": "Our results demonstrate that normal cfDNA results inconsistent with high-resolution abnormal ultrasounds should be confirmed by karyotyping following a substantial frequency of incorrect cfDNA results.The American College of Obstetrics and Gynecology (ACOG) and Maternal Fetal Medicine (MFM) Societies recommended that Historical review of our ~4,000 signed prenatal karyotypes found ~24% of reported abnormalities would not have been detected by cfDNA. Akron Children\u2019s Hospital Cytogenetics Laboratory has completed 28 abnormal cfDNA cases among the 112 amniocenteses karyotyped.normal newborn with a complex mosaic karyotype substantially decreasing the newborn\u2019s eventual reproductive fitness. This second case establishes the importance of\u00a0karyotyping the placenta and cord or peripheral blood when inconsistent or mosaic results are identified following an abnormal cfDNA result with a normal newborn phenotype without a prenatal karyotype.Following abnormal cfDNA results our karyotypes confirmed only 60% of the cfDNA results were consistent. Our cases found a normal cfDNA test result followed by a 20\u00a0weeks anatomical ultrasound detected a false negative trisomy 18 cfDNA result. One cfDNA result that reported trisomy 21 in the fetus was confirmed by karyotyping which also added an originally undetected balanced reciprocal translocation. Another reported karyotyped case followed by a repeated microarray of pure fetal DNA, together revealed one phenotypically These Maternal Fetal Medicine referrals demonstrate that positive NIPT results identify an increased abnormal karyotypic frequency as well as a substantial proportion of discordant fetal results. Our results found: (1) a normal NIPT test result followed by a 20\u00a0week anatomical ultrasound detected a false negative trisomy 18 NIPT result, (2) a substantial proportion of abnormal NIPT tests identify chromosomal mosaicism that may or may not be confined to the placenta, (3) follow up karyotyping should be completed on the newborn placenta and peripheral blood when the amniocyte karyotype does not confirm the NIPT reported abnormality in order to identify ongoing risk of developing mosaic symptoms, and (4) karyotyping all high risk fetuses tested by amniocentesis defines the 24% of chromosome abnormalities not currently screened by NIPT.The online version of this article (doi:10.1186/s12967-015-0569-y) contains supplementary material, which is available to authorized users. Interest in circulating cell free DNA fragments began when transplacental transmission of fetal cells into the maternal circulation was demonstrated by detection of fetal erythrocytes . Subsequa. This was selected as a preferred source of fetal DNA to be tested because it has an average half-life of 16.3\u00a0min post-delivery so that levels are undetectable within hours post-partum .Term placentas from positive testing fetuses were placed into medium, stored cold, and transported at 4\u00b0C prior to analysis. Pathologists Dr. Kimberly Eickholt or Dr. Dimitris Agamanolis obtained biopsied samples of villous rich tissue from each placenta. According to our placental Products of Conception (POC) protocol, cytogenetic technologists further dissected the chorionic villi from the maternal deciduas, disbursed the cells with collagenase, and seeded the cells in two independent flasks. The dissected chorionic villous sampled (CVS) cells that grew in two flasks were karyotyped. In addition, an aliquot of collagen digested villous cells were prepared in hypotonic and fixed for possible additional FISH analysis. Chorionic villous cells that grew in culture were karyotyped according to standard protocols.The placenta from the circulating placental DNA testing positive for Turner Syndrome Table\u00a0, case 5 Akron Children\u2019s Hospital Internal Review Board Chairman Dr. Robert Novak discussed and assured compliance with required clinical and research applications of these results at each step in the process. The patients\u2019 results reported in detail have a signed permission to publish these data.One hundred three 103) amniocentesis samples Table\u00a0 have bee03 amniocFive NIPT results reflecting fetal trisomy 18 were confirmed by follow up karyotypes of amniocytes. In contrast, one fetus was reported in an early test to be normal for all chromosomes tested by MaterniT21 including chromosome 18 with a specificity of 99.6% and a CI 99.2\u201399.6% for chromosome 18 NIPT results indicating each fetus had trisomy 21 were confirmed by follow up amniocyte karyotypes. The fifth patient was reported to be carrying a 16\u00a0week 5\u00a0day fetus positive for Trisomy 21 by Ariosa (q13q21.3)[41] (q21.1q21.3) does not modify patient prognosis. Given a single XIST gene locus on both the patient\u2019s normal and derivative X chromosomes without an extra XIST copy by FISH, this copy number minimally modify a female newborn\u2019s phenotype provided the duplication breakpoints did not result in an X-linked dominant genetic disease. This is in contrast to an X chromosome with two XIST genes that typically inactivate nearly all X chromosome genes to mimic a Turner phenotype . HoweverOne of five trisomy 21 patients tested by NIPT and confirmed to have trisomy 21 also had a balanced reciprocal translocation characterized by G-banded karyotyping: 46, XY, t5;10). Table\u00a0, 10 A foThese cases illustrate the following principles: (1) Noninvasive prenatal testing is enhanced by thorough ongoing patient counseling. (2) Abnormal ultrasounds with a recognizable genetic etiology should be offered an invasive procedure to test for a genetic abnormality even when placental DNA screening was normal. (3) The reported frequencies of abnormal circulating placental DNAs is lower than the reported ~1.5% mosaic CVS frequency, suggesting that many chromosomally mosaic placentas are not detected by NIPT. (4) Given that ~20% of reported placental mosaicism is also detectable in the fetus , amniocyOur initial reported karyotyped case demonstrated that fetuses with abnormal ultrasounds and a recognizable etiology should be offered an invasive diagnostic procedure to test for the most likely genetic origin of the phenotype. Improved ultrasound devices, training, and experience enable obstetricians to characterize abnormal and normal fetal phenotypes in spite of an erroneous test result. Thus judiciously interpreted ultrasounds provide an important check to any laboratory test result.Placental mosaicism has an overall frequency of ~1.5% in five large chorionic villus sample studies , 13\u201317. One of our trisomy 21 NIPT results completing invasive prenatal diagnosis of all abnormal ultrasounds in the face of existing normal placental DNA results, and (2) completing results on newborn bloods to either confirm a normal invasive prenatal karyotype is not mosaic in another tissue or to test the newborn karyotype when a prenatal karyotype was not obtained following an abnormal ultrasound.This manuscript 1) reports and explains the predicted increased frequency of discordant placental DNA results to fetal karyotypes at Maternal Fetal Medicine referral centers, (2) demonstrates the importance of follow up ultrasound with normal NIPT results, (3) karyotyping phenotypically normal newborns with abnormal NIPT results, (4) ongoing thorough counseling, and (5) provides our laboratory protocol for follow up analysis of placenta along with cord or peripheral newborn blood are attributed to the enrichment of positive testing trophoblastic cases referred to Maternal Fetal Medicine specialists. Our last case correctly reported monosomy X in a mosaic placenta, but found a different major chromosome abnormality in a phenotypically normal newborn predicting high risk of major unanticipated difficulties at reproductive age. Presented cases highlight our attention to (1) the abnormal subsequent ultrasound following a normal circulating trophoblastic DNA (cfDNA) result on a trisomy 18 fetus, (2) the substantial proportion of abnormal mosaic placental karyotypes among all abnormal cfDNA results that may or may not be identified in the fetus, (3) the correct fetal information in advance of patient care even when the patient elects to carry an abnormal fetus, and (4) aFetal DNA circulating in maternal blood originating in the placenta is referenced as the target of Noninvasive Prenatal Testing (NIPT) in the remainder of this study.bLicensed tests initially offered commercially, 2013:A.Harmony Prenatal Test\u2122 , employs directed analysis of cell-free DNA fragments. This test selected regions (DANSR\u2122) with a proprietary algorithm, FORTE\u2122, to selectively analyze cell-free DNA in maternal blood for trisomy 21, 18 and 13.The B.Verifi\u2122 Prenatal Test , utilizes massively parallel DNA sequencing to identify an increased representation of chromosomes 21, 18, and13. Verinata Health Inc also offers testing for Monosomy X (Turner Syndrome) as an additional option to the verifi(TM) prenatal test;The C.MaterniT21\u2122 Plus Test , utilizes massively parallel DNA sequencing to identify trisomy of chromosomes 21, 18, 13, 16, and 22; 22q deletion syndrome (DiGeorge), 5p (Cri-du-chat syndrome), 15q (Prader-Willi/Angelman syndromes), 1p36 deletion syndrome.D.Natera, Inc. 201 Industrial Road, Suite 410, San Carlos, CA 94070, (650) 249-9090] provides the Panorama prenatal test based upon single nucleotide polymorphisms.cThe 4 CVS samples were further analyzed by FISH for chromosome 21 and chromosome 13 copy number. Following collagenase digestion of dissected villous cells, hypotonic, and immediate fixation, FISH analysis found 50 of 50 cells with two copies of the chromosome 13q probe but 32% of these nuclei with three copies of the 21q probe . The other three biopsied CVS samples had 68, 40, and 80% trisomy 21 nuclei among 25 scored cells each while all nuclei had two copies of the control chromosome 13 locus."} +{"text": "Ischemic preconditioning induces complex physiological adaptations to improve the tolerance of cells and tissues to future ischemic episodes. This phenomenon has been most studied in myocardial cells but also occurs in other tissues.Ischemic preconditioning could be used to improve the microcirculatory response to sepsis related hypoperfusion.Sixteen adult sheep (24-34 Kg) were anesthetized , mechanically ventilated and invasively monitored. Abdominal sepsis was induced by injecting autologous feces into the peritoneal cavity. Animals were randomly allocated to undergo remote ischemic preconditioning followed by intermittent post-conditioning (PRECON) or not (CONTROL). Controlled ischemic episodes of the lower extremities and pelvis were obtained by inflating an intravascular balloon in the aortic bifurcation. We performed 4 cycles of 2 min ischemia (4 min apart) 1 hour before sepsis induction and, thereafter, 1 inflation every 4 hours until death. Sublingual microcirculation was evaluated using side-stream dark field (SDF) video-microscopy capturing 5 videos of 12 sec at baseline and every 6 hours thereafter for later blinded analysis. We calculated the perfused vessel density (PVD), proportion of perfused vessels (PPV), mean flow index (MFI) and heterogeneity of PPV (PPV HI). Animals were followed until death or for a maximum of 30 hours. Data are presented as median values with inter-quartile ranges. Repeated measurement data were analyzed using a Generalized Estimating Equations approach in SPSS 19.0 with a p < 0.05 considered as significant.See table Repeated ischemic pre- and post-conditioning of the pelvis and lower extremities protected the microcirculation in this sheep model of severe abdominal sepsis."} +{"text": "Aborted sudden cardiac death (SCD) is a common indication for CMR. No studies have investigated the prognostic value of late gadolinium enhancement (LGE) solely in patients with secondary prevention implantable cardioverter-defibrillators (ICD) or aborted SCD.58 consecutive patients who had CMR and ICD implantation after aborted SCD between 2006 and 2013 were identified. Clinical notes and CMR results were reviewed. Patients were scanned with a CMR protocol that included a short axis cine stack and LGE imaging. CMR images were analysed blinded to clinical data using standard software . Patients were followed up after ICD implantation at six weeks, three months and six months and then every six months. Appropriate ICD therapy was defined as any anti-tachycardia pacing (ATP) or shock therapy delivered for a ventricular arrhythmia.Follow up data was available on 55/58 patients. Four patients died during the follow up period and one patient was lost to follow up 14 days post-implantation. Median follow up was 791 days (interquartile range 370-1142 days). 11 patients had appropriate ICD therapy. 6 had ATP only and 5 had appropriate shocks \u00b1ATP for ventricular arrhythmias. The clinical aetiology of the aborted SCD was ischaemic heart disease (IHD) 24, primary arrhythmia 14, arrhythmogenic right ventricular cardiomyopathy 7, dilated cardiomyopathy 6, myocarditis 2 and other 5.Of 55 patients on whom clinical outcome data was available 20 patients had ejection fraction (EF)<35% and 35 had EF>35%. 5/20 (25%) of those with EF<35% had appropriate ICD therapies and 6/35 17%) of those with EF>35% had appropriate ICD therapies on follow up. There was no significant difference in the likelihood ratio (LR) of appropriate therapy using an EF cutoff of 35% , confirmed on log rank analysis of Kaplan Meier curves LGE +ve patients had appropriate ICD therapies on follow up and only 1/20 (5%) LGE-ve patients had appropriate ICD therapies. LGE +ve patients were significantly more likely to have an appropriate ICD therapy on follow up , confirmed on log rank analysis of Kaplan Meier curves (P=0.036).In patients who have suffered aborted SCD, CMR is able to identify patients at risk of future arrhythmic events according to the presence or absence of LGE.PPS (FS/12/88/29901) and SP (FS/10/62/28409) are funded by British Heart Foundation fellowships."} +{"text": "MicroRNAs play an important role in cardiac remodeling. MicroRNA 499 (miR499) is highly enriched in cardiomyocytes and targets the gene for Calcineurin A (CnA), which is associated with mitochondrial fission and apoptosis. The mechanism regulating miR499 in stretched cardiomyocytes and in volume overloaded heart is unclear. We sought to investigate the mechanism regulating miR499 and CnA in stretched cardiomyocytes and in volume overload-induced heart failure.Rat cardiomyocytes grown on a flexible membrane base were stretched via vacuum to 20% of maximum elongation at 60 cycles/min. An in vivo model of volume overload with aorta-caval shunt in adult rats was used to study miR499 expression. Mechanical stretch downregulated miR499 expression, and enhanced the expression of CnA protein and mRNA after 12 hours of stretch. Expression of CnA and calcineurin activity was suppressed with miR499 overexpression; whereas, expression of dephosphorylated dynamin-related protein 1 (Drp1) was suppressed with miR499 overexpression and CnA siRNA. Adding p53 siRNA reversed the downregulation of miR499 when stretched. A gel shift assay and promoter-activity assay demonstrated that stretch increased p53 DNA binding activity but decreased miR499 promoter activity. When the miR499 promoter p53-binding site was mutated, the inhibition of miR499 promoter activity with stretch was reversed. The in vivo aorta-caval shunt also showed downregulated myocardial miR499 and overexpression of miR499 suppressed CnA and cellular apoptosis.The miR499-controlled apoptotic pathway involving CnA and Drp1 in stretched cardiomyocytes may be regulated by p53 through the transcriptional regulation of miR499. Oligonucleotide sequences of p53 were the consensus 5\u2019-AGTATGGTCTAGCCTGGCCC-3\u2019. The mutant oligonucleotides sequence was 5\u2019-AGTATGGTACCACCTGGCCC-3\u2019. After the p53 was radiolabelled, the nuclear extracts (4\u03bcg of protein in 2\u03bcL of nuclear extract) were mixed with 20pmol of the appropriate [\u03b332-p]ATP-labelled consensus or mutant oligonucleotides in a total volume 20\u03bcL for 30 min at room temperature. The samples were then resolved on a 4% polyacrylamide gel. Gels were dried and imaged using autoradiography. A control was performed in each case with mutant or cold oligonucleotides to compete with the labeled sequence.Nuclear protein concentrations from cultured cardiomyocytes were determined using a protein assay . Consensus and control oligonucleotides were labeled using polynucleotide kinase incorporation of [\u03b3A bp -941 to -442 rats miR499 promoter construct was generated as follows. Rat genomic DNA was amplified with forward primer 5\u2019-ATAACGCGTAGGAATCTCCCCCTCT-3\u2019 and reverse primer 5\u2019-CCTAGATCTGTAAGTGATGGTCGTCC-3\u2019. The amplified product was digested with Mlul and Bglll restriction enzymes and ligated into pGL3-basic luciferase plasmid vector . The miR499 promoter contained p53 conserved sites (CTAG) at -694 to -690bp. Rat CnA promoter genomic DNA was amplified with forward primer 5\u2019-TGACGCGTGTTTAATCCATCTCTGTTGG-3\u2019 and reverse primer 5\u2019-TGAGATCTCTTCTATCTGGCAAAGAAATTTTA-3\u2019. The CnA promoter contained miR499 conserved sites (AAGCAGTCATGCAATGGCTTAA) at 908 to 929 bp. For the mutant, the p53 binding sites in miR499 promoter and miR499 binding site in CnA promoter were mutated using a mutagenesis kit . Site-specific mutations were confirmed using DNA sequencing. Plasmids were transfected into cardiomyocytes using a low pressure-accelerated gene gun per the manufacturer\u2019s protocol (Bioware Technologies). Briefly, 2\u03bcg of plasmid DNA was suspended in 5\u03bcL of PBS and was delivered to the cultured cells at a helium pressure of 15psi. The transfective efficiency using this method was approximately 30%. The cell extracts were prepared using a dual luciferase reporter assay system and measured for dual luciferase activity with a luminometer . Renilla luciferase activity was normalized to firefly luciferase activity.Cell viability after the application of cyclic stretch was monitored using a trypan blue staining procedure and the 3--2,5-diphenyl tetrazolium bromide (MTT) assay to detect for stretch-induced cell injury. Cytotoxicity studies were performed as previously described.Apoptotic cells were quantified as the percentage of cells with hypodiploid DNA (sub-G1). Cardiomyocytes were fixed with 70% ethanol and treated with RNase. Nuclei were then stained with propidium iodide and fluorescein isothiocyanate\u2013annexin V. Cardiomyocytes that were negative for both annexin V and propidum iodide were considered to be alive. Cells that were positive for annexin V and negative for propidium iodide were considered to be undergoing apoptosis. Cells that were positive for both annexin V and propidium iodide were considered to be in the end stage of apoptosis, called second apoptosis. DNA content was measured using a FACSCalibur flow cytometer and Cell Quest software . Ten thousand cells were counted in all assays.Cellular ATP levels and protease activity were measured after a 90-minute incubation using the Mitochrondrial ToxGlo assay as per manufacturer\u2019s instructions. These 2 sets of data were combined to represent mitochrondrial dysfunction-related cytotoxic mechanisms. Apoptosis was also determined via caspase-3 activity using the caspase-3 colorimetric activity assay kit . The stretched cardiomyocytes were resuspended in chilled 1X Cell Lysis buffer and then centrifuged for 5 mins (10000 x g). The supernatant was then transferred to a fresh tube, adding 5X assay buffer and caspase-3 substrate. After incubating for 2 hours at 37\u00b0C, the sample was evaluated at 405 nm in a microtiter plate reader.DNA fragmentation was determined via TUNEL assay using the ApopTag peroxidase in situ apoptosis detection kit . The methodology for the TUNEL assay is further described in detail in the The methodologies for the immunohistochemical analysis and in situ hybridization assay in the AV shunt rat model are further described in detail in the All results are expressed as means \u00b1 SEM. Statistical significance was evaluated using analysis of variance . Dunnett\u2019s test was used to compare multiple groups with a single control group. The Turkey\u2013Kramer comparison was used for pair-wise comparisons between multiple groups following the ANOVA. A value of P < 0.05 was considered as significant.To investigate the effect of mechanical stretch on miR499 expression in cardiomyocytes, TaqMan MicroRNA real-time quantitative PCR (Applied Biosystems) was used to measure miR499. When cardiomyocytes were stretched to 10% of elongation, miR499 levels were similar to those of control cells (no stretch). Mechanical stretch of 20% for 2 hours resulted in significant miR499 expression compared with control cells, however stretch for 4 to 12 hours significantly down regulated miR499 expression comparedOverexpression of miR499 significantly inhibited CnA expression and CalcMiR499 regulation of Drp1 accumulation in cardiomyocytes through its effects on CnA expression was also examined. The expression of unphosphorylated (Drp1) and phosphorylated (p-Drp1) were induced by mechanical stretch of 20% for 8 hours . MechaniThe involvement of p53 in the regulation of miR499 expression in mechanical stretch was examined. p53 expression was induced by mechanical stretch of 20% for 4 to 12 hours . The expMechanical stretch of cardiomyocytes for 2 to 8 hours increased p53 DNA binding activity . To studApoptosis was assessed using propidium iodide-annexin V double staining and FACS analysis. The percentage of cells stained with annexin V was elevated following stretch for 8 hours . The obsThe influence of miR499 on the expression of CnA in vivo was examined. AV shunt was performed in adult Sprague-Dawley rats to induce volume overload. Heart weight and heart weight/body weight ratio significantly increased for 14 and 28 days after AV shunt . The heaAV shunt significantly increased myocardial miR499 expression from 1 day to 5 days after shunting, but later decreased significantly from 7 days to 28 days compared to AV shunt at 5 days . These fTo investigate the effect of miR499 on myocardial CnA expression, overexpression of miR499, antagomir-499, and mutant type miR499 (mut-499) in the left ventricle was performed. AV shunt at 14 days significantly increased myocardial CnA protein expression . The carAt 14 days after AV shunt, the presence of miR499 in cardiomyocyte cytoplasm was confirmed using in situ hybridization . ImmunohA significant increase in TUNEL positive nuclei was present in AV shunt hearts . IncreasThe heart weight, heart weight/body weight ratio, LVEDD and LVESD were significantly improved after overexpression of miR499. The fraction shortening (FS) of LV was also improved but statically insignificant. Adding antagomir 499 attenuated the effect of miR499, while mutant miR499 have no effect on heart size after AV shunt. We currently report the following major findings: (i) mechanical stretch, as well as AV shunt, down regulates miR499 in cardiomyocytes; (ii) mechanical stretch of cardiomyocytes induces the expression of CnA at the protein and mRNA levels, and cellular activity of calcineurin; (iii) expression of CnA and calcineurin activity are suppressed by miR499 during stretch; (iv) when exposed to mechanical stretch, dephosphorylated Drp1 expression is suppressed by overexpression of miR499 and CnA siRNA; (v) mechanical stretch induces apoptosis of cardiomyocytes via an miR499-controlled apoptotic pathway involving CnA and Drp1; (vi) suppression of p53 reverses the transcriptional downregulation of miR499 by mechanical stretch; (vii) CnA and myocardial cellular apoptosis were induced in heart failure induced by volume overload; and (viii) overexpression of miR499 suppressed CnA and attenuated myocardial cellular apoptosis after AV shunt.Mechanical stretch triggers cardiomyocyte remodeling characterized by loss of contractile tissue, hypertrophy, and increased fibrotic tissue. Several We currently demonstrate significant miR499 down regulation with mechanical stretch and volume overload. This down-regulation results in increased CnA and expression of dephosphorylated Drp1due to mitochondrial fission and cellular apoptosis. Overexpression of miR499 reduced CnA and dephosphorylated Drp1 protein levels, whereas application of antagomir-499 caused a robust increase in CnA and consequent dephosphorylated Drp1 protein, indicating a relief of tonic repression of CnA and dephosphorylated Drp1 by miR499. Taken together, these results suggest that miR499 plays an essential role in the synthesis of CnA and dephosphorylated Drp1 when placed under mechanical stretch and may provide important new information on the role of miR499 in CnA-Drp1\u2013mediated apoptosis.In the present study, while miR499 was down regulated 4 to 12 hours after mechanical stretch, CnA and dephosphorylated Drp1 protein levels were consistently increased after 6 to 12 hours of stretch. These findings are consistent with previous observations that CnA expression and activity of calcineurin increases under physiological stressful conditions, including heart failure and hypoMiR499 expression may be down regulated by mechanical stretch. The level of p53 is increased after mechanical stretch and is a modulator of cardiomyocyte apoptosis after stretch., 43 ThesThe promoter activity analysis verified the regulation of miR499 expression at the transcriptional level when cardiomyocytes were placed under mechanical stretch. Mechanical stretch increased the binding activity of p53 to DNA and the removal of the p53-binding site in the miR499 promoter area abolished the suppression effect of p53 on miR499 promoter activity. These finding suggest the decreased transcriptional activity of the miR499 promoter due to mechanical stretch is depended on p53. Therefore, it is likely that enhanced p53 activity associated with mechanical stretch down-regulates miR499 transcription, which results in relief of repression of CnA, and thereby an increase in dephosphorylated Drp1, leading to mitochondrial fission and cardiomyocytes apoptosis. In addition, the current report is supported by Liao et al. who demonstrated that p53 is upregulated at relatively late time after mechanical stretch. The regu2O2-induced injury via its effects on Pdcd4 and Pacs2.[Previous study by Wang J et al had demonstrated that miR499 protects cardiomyocytes from Hnd Pacs2. In the pThe current study is the first to demonstrate a link between p53, miR499, CnA, and Drp1 and cardiomyocyte apoptosis when placed under mechanical stretch. CnA is a direct target of miR499 and miR499 inhibits cardiomyocyte apoptosis through the suppression of CnA-mediated Drp1, thereby decreasing Drp1-mediated activation of mitochondrial fission. We also observed the down regulation of miR499 expression by p53 transcription when placed under mechanical stretch. Overexpression of miR499 attenuated cardiomyocyte apoptosis in heart failure induced by volume overload. Modulation of miR499 levels could provide a therapeutic approach for treating heart failure.S1 FigThe green spot is miR499 in situ image.(TIF)Click here for additional data file.S2 Fig(TIF)Click here for additional data file.S3 Fig* p < 0.001 vs. control. \u00a7 p < 0.001 vs. stretch alone.(TIF)Click here for additional data file.S4 Fig(TIF)Click here for additional data file.S5 FigRepresentative microscopic images showing the presence of miR499 (green color) in the cytoplasm of cardiac myocytes from left ventricular myocardium in AV shunt rats. The sham groups or scrambled probe did not detect the presence of miR499.(TIF)Click here for additional data file.S6 FigTUNEL staining is indicative of cell death.(TIF)Click here for additional data file.S1 File(DOC)Click here for additional data file.S1 Table(DOC)Click here for additional data file."} +{"text": "Mesp family proteins comprise two members named mesodermal posterior 1 (Mesp1) and mesodermal posterior 2 (Mesp2). Both Mesp1 and Mesp2 are transcription factors and they share an almost identical basic helix-loop-helix motif. They have been shown to play critical regulating roles in mammalian heart and somite development. Mesp1 sits in the core of the complicated regulatory network for generation of cardiovascular progenitors while Mesp2 is central for somitogenesis. Here we summarize the similarities and differences in their molecular functions during mammalian early mesodermal development and discuss possible future research directions for further study of the functions of Mesp1 and Mesp2. A comprehensive knowledge of molecular functions of Mesp family proteins will eventually help us better understand mammalian heart development and somitogenesis as well as improve the production of specific cell types from pluripotent stem cells for future regenerative therapies. In humans and mice, Mesp family proteins comprise two members named mesodermal posterior 1 (Mesp1) and mesodermal posterior 2 (Mesp2), which have been shown to play critical roles in embryonic mesodermal development. Both Mesp proteins are transcription factors and share an almost identical basic helix-loop-helix (bHLH) motif studies showed that, in mouse embryonic development, the Mesp1-expressing cells were observed to ingress through the primitive streak and later in a wedge-shaped distribution in the nascent mesoderm at embryonic day 6.5\u20137.0 (E6.5\u20137.0) , and repressed genes involved in early primitive streak formation and endoderm specification . Additionally, chromatin immunoprecipitation experiments showed that Mesp1 directly bound to conserved E-Box within genomic regions of several upregulated and downregulated genes , suggesting that Mesp1 directly regulates gene transcription to induce cardiovascular specification and inhibit acquisition of other possible cell fates during this developmental stage cells derived from differentiated ESCs, compared with KDR+ PDGFR\u03b1+ cardiac progenitors in which no enrichment was observed into downstream cells of the 3 embryonic germ layers, although lacking of partial Moreover, significant effort has been made over the past two decades for direct reprogramming of fibroblasts into cardiomyocytes which are required for cardiac regenerative therapies. Recent studies showed that the efficiency for direct reprogramming of fibroblasts into cardiomyocytes could be strongly enhanced with the addition of Mesp1 and contains a unique region at the carboxyl-terminus . At the initial phase of Mesp2 expression in somitogenesis, Mesp2 exhibited a partially co-expressing pattern with NICD as reflected by double immunohistochemistry from the pluripotent stem cells for future cellular therapy and drug screening."} +{"text": "Pressure ulcer trials often use development of category\u22652 pressure ulcers as an endpoint. ResearcPressure ulcer monitoring systems have been introduced in the English NHS including the Safety Thermometer. A project, funded by the Tissue Viability Society and supported by NHS England, comprised a pressure ulcer/wound audit to assess the accuracy of current monitoring systems. The pressure ulcer/wound audit was conducted on the Safety Thermometer census date October 2014) in a stratified random sample of Trusts providing in-patient services in England, using \u2018gold-standard\u2019 prevalence methods[014 in a The results show low accuracy of routine pressure ulcer data sources; the weighted sensitivity estimate(95% CI) for the safety thermometer was 48.2%(35.4%-56.7%). The pressure ulcer/wound audit indicated that pressure ulcers reported on monitoring systems may not be readily obtained from clinical notes. These results support published data comparing ward and research nurse data. Questio"} +{"text": "These changes cause hypercapnic vasodilation and hypocapnic vasoconstriction, which affect the myocardial blood supply and subsequently myocardial oxygenation.Oxygenation-sensitive (OS) imaging relies on alterations of deoxyhemoglobin (dHb) in the blood. Thus signal intensity (SI) is dependent on the factors that shift the dHb fraction; oxygen saturation, tissue oxygen extraction and blood supply. We aimed to physically measure and compare these parameters while manipulating myocardial oxygenation with systemic CO2 = 100 mmHg). Arterial pCO2 levels of 30, 40 and 50 mmHg were targeted through ventilation settings. Coronary flow was measured using a surgically implanted perivascular MRI-compatible ultrasound flow probe around the left anterior descending (LAD) coronary artery. Blood gases were analyzed from blood drawn through a catheter in the femoral artery and in the coronary sinus, which were obtained simultaneously with LAD flow measurements and OS images. The images were analyzed in systole and assessed for the global myocardial %-change SI in comparison to the baseline values at paCO2 = 40 mmHg.In 8 anaesthetized swine, we acquired OS images in 3 short axis slices using an ECG triggered balanced SSFP sequence with standard cine imaging for function acquired in a short-axis stack in a clinical 3T magnet during normoxia (paO2 = 50 mmHg) in comparison to hypocapnia (paCO2 = 30 mmHg, Figure OS-SI increased significantly along with LAD flow during hypercapnia (paCO2-induced vasomotor response in combination with oxygenation-sensitive imaging of the myocardium.As oxygen extraction remained consistent during our maneuvers, the changes in OS-SI are explained by the increase in coronary blood flow, thus demonstrating the potential of COFunding is provided by the Montreal Heart Institute Foundation and the Canadian Foundation for Innovation."} +{"text": "In fact 6.3% of oil palm is commercially redundant (with negative NPV of $-299/ha\u2212yr-$-65/ha\u2212yr) due to palm mortality from flood inundation. These areas would have been important riparian or flooded forest types. Moreover, 30,173 ha of unprotected forest remain and despite its value for connectivity and biodiversity 64% is allocated for future oil palm. However, we estimate that at minimum 54% of these forests are unsuitable for this crop due to inundation events. If conversion to oil palm occurs, we predict a further 16,207 ha will become commercially redundant. This means that over 32,000 ha of forest within the floodplain would have been converted for little or no financial gain yet with significant cost to the ecosystem. Our findings have globally relevant implications for similar floodplain landscapes undergoing forest transformation to agriculture such as oil palm. Understanding landscape level constraints to this crop, and transferring these into policy and practice, may provide conservation and economic opportunities within these seemingly high opportunity cost landscapes.Lowland tropical forests are increasingly threatened with conversion to oil palm as global demand and high profit drives crop expansion throughout the world\u2019s tropical regions. Yet, landscapes are not homogeneous and regional constraints dictate land suitability for this crop. We conducted a regional study to investigate spatial and economic components of forest conversion to oil palm within a tropical floodplain in the Lower Kinabatangan, Sabah, Malaysian Borneo. The Kinabatangan ecosystem harbours significant biodiversity with globally threatened species but has suffered forest loss and fragmentation. We mapped the oil palm and forested landscapes and undertook economic modelling. Within the study region , 250,617 ha is cultivated with oil palm with 77% having high Net-Present-Value (NPV) estimates ($413/ha Elaeis guineensis) which drives biodiversity loss Lowland tropical forest ecosystems contain some of the highest levels of species endemism and biological diversity globally Elaeis guineensis is flood intolerant with mortality occurring from root rot within two weeks of ground saturation and/or low oxygen levels or from saline water Oil palm plantations are limited to low elevation areas and are in direct conflict with tropical lowland forests, including those found within riverine floodplains. Riverine floodplains are characterised by high levels of biodiversity and productivity We investigated forest and oil palm dynamics within a floodplain system in the Malaysian context. In Malaysia it is estimated that between 1990\u20132005, 55\u201359% of Malaysia\u2019s oil palm extent replaced old-growth and secondary forests The study area comprised 520,269 ha of the Lower Kinabatangan floodplain region, in eastern Sabah . The regwww.usgs.gov). Images were orthorectified and registered and indices calculated for: Normalized Difference Vegetation Index (NDVI); Normalized Difference Moisture Index (NDMI); and Soil Adjusted Vegetation Index (SAVI). Spectral transformations were performed including reflectance for SPOT5 and Tasseled Cap transformation on the Landsat TM5 Images Forest vegetation types were identified using SPOT5 10 m satellite images captured on 25/11/2007 and two on 19/06/2009; as well as two Landsat TM5 30 m images captured on 27/07/2006 and 11/08/2009 The eastern and western classifications were exported as shapefiles and \u2018merged\u2019 to form one continuous layer. An accuracy assessment was performed using an error confusion matrix method using the second portion of the ground truth dataset, and overall classification accuracy and kappa statistics were calculated for both portions Oil palm age and productivity classes were digitised using the 2.5 m SPOT5 images. Age classes were defined by characteristics viewable in the images that were associated with planting preparation and/or age associated features (see http://earth.google.com) in our region to validate our oil palm age and productivity map. Although our satellite imagery was 2.5 m, which allowed individual palms to be seen, the higher resolution tiles in Google Earth enabled better interpretation ability and therefore permitted the validation of classes. We defined these tiles within a GIS (ArcGIS 10), generated 120 random points high-resolution tiles from Google Earth images of oil palm for four oil palm class scenarios (see Tables S1 (a+b) to S4 (a+b) in \u221225. For all other models, we vary annual yields through the 25 year model timeframe, by calculating the proportion of yield (in t/FFB/ha) needed against our maximum \u2018Full stand\u2019 (100% at 136 SPH) yield values. In our models we assume a direct relationship between the number of SPH (or percentage thereof) and yield. We discuss the pitfalls and limitations of these assumptions in the discussion.Our economic models assumed a typical 25 year crop life as per industry standard Within our models we retained FFB as our revenue unit rather than converting to crude palm oil (CPO) as for smallholders and commercial estates with no processing mills, revenue is derived from the sale of FFB. We based our model largely around 2011 data. As a result we used a constant FFB price of US$178 t/FFB, based on average 2011 values from the east coast of Sabah . We used a discount rate of 11% per annum in our main models, used by industry in 2011 in Sabah. However, to assess model robustness we undertook sensitivity analyses using discount rates of 5%, 8% and 14% per annum. Cost data were calculated from estates past actual costs, their budgeted future costs and typical industry average costs for 2011 . These costs were cross referenced and supplemented with state wide data on costs from 2008 To predict suitable and unsuitable areas for oil palm outside the protected areas, we used a Classification and Regression Tree (CART) analysis. The CART framework was developed by using data from existing oil palm blocks integrated with biophysical spatial information developed at 1 ha resolution. Predictor variables included: (1) Euclidian distance from main river (and main tributaries); (2) elevation; (3) aspect; (4) slope; and (5) a soil agriculture suitability layer using 4 suitability classes n\u200a=\u200a14) used by commercial land valuation agencies and industry were used to forecast future loss of unprotected forest by quantifying the extent of alienated land. Maps were undated but known to have been drafted in the 1990\u2019s by the Land and Survey Department. Although, these maps have been updated since the 1990\u2019s the extent of alienated land is likely underestimated. Maps were scanned at high resolution and geo-referenced to 2.5 m SPOT5 images. Land parcels were digitised, with title types and identity numbers recorded in a vector file.Publicly available cadastral maps (Land title types included: (1) Native Title (NT), i.e., smallholdings alienated for agriculture for perpetuity and restricted to <40 ha in size mean size\u200a=\u200a679 ha; sd (standard deviation) \u200a=\u200a5,231 ha). Of the forested extent, 12% occurs outside the protected areas (i.e. unprotected forest).Overall classification accuracy was 72.2%, with a Kappa statistic of 0.65 using an error matrix method and 969 ground truth points ; and 51% under Country Land or commercial titles ; the remaining extent was State land (36%) see . Of the Overall classification accuracy was 84% with a Kappa statistic of 0.816 or new reforestation projects requires \u2018river reserves\u2019 to be retained in cultivated areas (up to 20 m from watercourse). This is rarely implemented by companies or enforced by authorities. River reserve widths are based on minimum needs to mitigate soil and bank erosion In this study we highlighted prevalent issues pertaining to forest conversion to oil palm in a floodplain system. Our study provides an exemplary case study of how poor planning can result in unfavourable land cover whilst little serving either profitability for landholders or conservation agendas. Regional studies such as this one should be conducted for other floodplains undergoing land cover change. Greater understanding of opportunities and constraints in these landscapes is needed to promote informed trade-off decision making at multi-scale levels. As global palm oil demand increases, ensuring responsible utility of tropical landscapes is vital in synergising a balancing between agricultural and development needs with long-term biodiversity conservation and ecosystem functioning. This study has global significance beyond our study region as we hypothesis that biophysical criteria used by many governments and international agencies for targeting areas for oil palm cultivation are likely similar to those used in Sabah File S1Oil palm age and productivity map validation areas ; Land ordinance policy under land titles (Appendix S1); Operational habitat map confusion matrix (Appendix S2); Confusion matrix table of the OBIA (Table S6); Confusion matrix table of the oil palm age and productivity (Table S7); Decision Tree from the CART analysis for oil palm suitability .(PDF)Click here for additional data file.File S2Excel economic models for four oil palm classes including: Full stand 100% palm capacity (136 SPH) (Table S1(a)); Full stand at 76% palm capacity (103 SPH) (Table S1(b)); Underproductive at 75% at 75% capacity (102 SPH) (Table S2(a)); Underproductive at 75% at 51% capacity (69 SPH) (Table S2(b)); Underproductive at 50% at 50% capacity (68 SPH) (Table S3(a)); Underproductive at 50% at 26% capacity (35 SPH) (Table S3(b)); Underproductive at 25% at 25% capacity (34 SPH) (Table S4(a)); Underproductive at 25% at 0% capacity (0 SPH) (Table S4(b)); Discounted summaries (Table S5); Yield curve . The authors are solely responsible for the content and functionality of these materials. Queries should be directed to the corresponding author.(XLSX)Click here for additional data file."} +{"text": "AngII and TGF-\u03b2 interact in development of thoracic and abdominal aortic diseases, although there are many facets of this interaction that have not been clearly defined. The aim of the present study was to determine the effects of TGF-\u03b2 neutralization on AngII induced-aortic pathologies. Male C57BL/6J mice were administered with either a rabbit or mouse TGF-\u03b2 neutralizing antibody and then infused with AngII. The rabbit TGF-\u03b2 antibody modestly reduced serum TGF-\u03b2 concentrations, with no significant enhancements to AngII-induced aneurysm or rupture. Administration of this rabbit TGF-\u03b2 antibody in mice led to high serum titers against rabbit IgG that may have attenuated the neutralization. In contrast, a mouse TGF-\u03b2 antibody (1D11) significantly increased rupture in both the ascending and suprarenal aortic regions, but only at doses that markedly decreased serum TGF-\u03b2 concentrations. High doses of 1D11 antibody significantly increased AngII-induced ascending and suprarenal aortic dilatation. To determine whether TGF-\u03b2 neutralization had effects in mice previously infused with AngII, the 1D11 antibody was injected into mice that had been infused with AngII for 28 days and were observed during continued infusion for a further 28 days. Despite near ablations of serum TGF-\u03b2 concentrations, the mouse TGF-\u03b2 antibody had no effect on aortic rupture or dimensions in either ascending or suprarenal region. These data provide further evidence that AngII-induced aortic rupture is enhanced greatly by TGF-\u03b2 neutralization when initiated before pathogenesis. Aortic aneurysm is defined as a permanent dilation of the lumen and represents a potentially fatal condition due to its high risk for rupture . Aortic AngII-induced aortopathies have been associated with regulation of TGF-\u03b2 signaling. TGF-\u03b2 is a multifunctional cytokine that may mediate aortic diseases through complex pathways in both thoracic and abdominal regions ,14. DeveUnlike antagonism of AngII, the effects of neutralizing TGF-\u03b2 activity on aortopathies have been variable. In thoracic aortas, the effects of TGF-\u03b2 neutralization have ranged from reducing aneurysms in fibrillin1 haploinsufficient mice and AngIet al. [et al. [AngII and TGF-\u03b2 interact in the development of aortopathies, although there are many uncertainties regarding their mechanisms. These uncertainties include the variable effects on: 1. aortopathies in the thoracic versus abdominal area; 2. aortic dilatation versus rupture; 3. effects on aortic disease when neutralization was introduced after prolonged AngII infusion. Wang et al. administ [et al. . Using tEight week old, male C57BL/6J mice were purchased from The Jackson Laboratory . Mice were group housed in individually vented cages with negative air exhaust pressure on a light:dark cycle of 14:10 hours. Rodent bedding consisted of Sani-Chip . Mice were fed a normal laboratory rodent diet and provided with drinking water from a reverse osmosis system.The recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health were followed. All procedures were approved by the Institutional Animal Care and Use Committee of the University of Kentucky . Study mice were observed daily for normal behavior and weighed weekly throughout experimental duration. Humane endpoints included deteriorating body condition score, weight loss of >15%, inability to rise or ambulate, dyspnea, and dehydration. None of the study mice exhibited any of the humane endpoint criteria. All surgeries were performed under isoflurane anesthesia and post-surgery care utilized local topical analgesia for minimizing stress and pain. Aortic rupture may occur in the AngII-infusion aneurysm mouse model . In thisAngII was infused via mini-osmotic pumps subcutaneously implanted on the flank of mice .For the first study, mice were injected intraperitoneally at a dose of 10 mg/kg twice a week with either control rabbit nonimmune IgG antibody , or polyclonal rabbit TGF-\u03b2 IgG antibody . One week of IgG injections were given prior to the initiation of AngII infusion and the twice weekly injections were continued throughout the 28 day infusion of AngII .1 antibody or mouse IgG1 isotype control at a range of doses for 1 week. The dose of monoclonal TGF-\u03b2 IgG1 antibody and mouse IgG1 isotype control in the subsequent study was selected based on the outcome of the dosage experiment . One week of IgG injections were given prior to AngII infusion, and injections were continued throughout the 28 day infusion of AngII were injected into mice after 28 days of AngII infusion. During the period of antibody injections, mice were infused continuously with a new pump filled with AngII for an additional 28 days . Serum samples were acidified with HCl (1 M) for 10 minutes to activate latent TGF-\u03b21, and then neutralized with NaOH (0.5 M)/HEPES (1.2 N).To examine whether rabbit IgG antibodies promoted an immune response to stimulate production of mouse anti-rabbit antibodies, serum concentrations of mouse anti-rabbit IgG titers were measured by ELISA. Sera from mice without antibody injection were used as negative controls. Plates were coated with normal rabbit IgG diluted in PBS and incubated overnight in a humidifying chamber. Serial dilutions of sera from study mice diluted in BSA-PBS (1% wt/vol) were added to the wells and incubated for 2 hours. Goat horseradish peroxidase (HRP) conjugated antimouse Ab was added and incubated for 30 minutes. Chromogen ABTS was added to each well and read at 450 nm using a microplate reader.Cell numbers in peripheral blood were counted using a Hemavet 950 (Drew Scientific Inc).TM transducer . The resolution of this 18\u201338 MHz transducer is approximately 50 \u03bcm. Mice were anesthetized using IsofluraneTM . Chest and abdominal hair was removed using clippers and depilatory cream . Ultrasonic gel was placed on the area to be scanned. Two-dimensional images (B mode) of short-axis scans were acquired to measure the maximal diameter of suprarenal aortas and long-axis scans for maximal diameter of ascending aortas at selected intervals . For two groups, comparisons were performed using an unpaired two-tailed Student\u2019s t test for normally distributed continuous variables and a Mann-Whitney U test for non-normally distributed variables. One-way ANOVA with Student-Newman-Keuls was used to test a variable of more than 2 groups. A Fisher\u2019s exact test was used to compare the incidence of aortic rupture. Two way ANOVA with Holm-Sidak post hoc analysis was performed for multiple-group and multiple-manipulation analyses. Kaplan-Meier survival curves were constructed and analyzed using log-rank (Mantel-Cox) test. P<0.05 was considered statistically significant.To determine the effects of TGF-\u03b2 neutralization on ascending and suprarenal aortic aneurysm formation, we injected either IgG control or polyclonal rabbit TGF-\u03b2 antibodies, at a dose of 10 mg/kg twice per week, into C57BL/6J mice infused with AngII. Injections were started 1 week prior to infusion of AngII. Injection of the rabbit TGF-\u03b2 antibody had no overt detrimental effects on mouse health and body weight. Plasma renin was measured as an index of AngII potency, and as expected, was decreased significantly during AngII infusion , Study 1A possible basis for the minimal effects of this antibody was attributable to the injection of a rabbit protein stimulating production of mouse anti-rabbit antibodies, thereby decreasing biological efficacy of this rabbit polyclonal TGF-\u03b2 antibody. Therefore, we measured mouse antirabbit IgG antibodies using an ELISA method. Mice injected with the rabbit polyclonal TGF-\u03b2 antibody had high serum titers of anti-rabbit antibodies. This immune response may contribute to the minor effects observed in the rabbit TGF-\u03b2 antibody injected group .To overcome a potential confounding immune response of injecting a cross-species antibody, we performed further studies using a mouse TGF-\u03b2 neutralizing antibody (1D11) to examine its effects on AngII-induced ascending and suprarenal aortic aneurysms in C57BL/6J mice. First, we determined a dose of 1D11 that produced sustained reductions in serum TGF-\u03b2 concentrations. Injections of 1D11 at 5 mg/kg, 3 times a week, substantially reduced serum TGF-\u03b2 concentrations in 1 week . In subsTo examine whether there were any early changes in aortic dimensions during TGF-\u03b2 neutralization, we used high-frequency ultrasound to measure diameters of ascending and suprarenal aortas prior to, and 4 days after AngII infusion in mice injected with either 1D11 or isotype-match IgG control. Although AngII promoted lumenal dilation in both aortic regions, 1D11 had no effects on early ascending or supraTo determine whether TGF-\u03b2 neutralization had dose-dependent divergent effects on aortopathies, we compared the effects of low and high doses of 1D11 on AngII-induced aortic aneurysms. Both low and high doses of 1D11 had no effect on body weight and platelet count , Study 31 control into mice for a further 28 days. During the additional 28 day period of antibody injections, mice were continuously infused with AngII. As with previous studies, injection of 1D11 had no effect on body weight or platelet count ascending aortic and (B) abdominal aortic rupture in mice infused with AngII. Arrows point to thrombi which are dark red in color.(PDF)Click here for additional data file.S2 FigArrows denote single time points. Red box denotes continuous infusion in vivo. AngII = Angiotensin II . Ctrl = Isotype-matched control IgG.(PDF)Click here for additional data file.S3 FigArrows denote single time points. Red box denotes continuous infusion in vivo. AngII = Angiotensin II . Ctrl = Isotype-matched control IgG.(PDF)Click here for additional data file.S4 FigArrows denote single time points. Red box denotes continuous infusion in vivo. AngII = Angiotensin II . Ctrl = Isotype-matched control IgG.(PDF)Click here for additional data file.S5 FigA. The ascending area of the ascending aorta is indicated by tracing in teal. Diameters of proximal ascending aortas were measured (yellow line). Yellow asterisks indicate the innominate, common carotid and subclavian arterial branches of the aorta. B. The suprarenal abdominal aorta is indicated by tracing in teal. Diameters of suprarenal abdominal aortas were measured (yellow line).(PDF)Click here for additional data file.S6 FigRepresentative photographs of saline and AngII-infused abdominal aortas. Aortas are harvested from mice, fixed overnight in formalin, cleaned of surrounding tissues, and cut in half at the diaphragm. The abdominal portions are pinned and photographed. Black lines depict maximal diameter measurements of suprarenal aortas. Numbers in white box are actual measurement in mm.(PDF)Click here for additional data file.S7 FigAortas were harvested from mice, fixed overnight in formalin and cleaned of surrounding tissue. Thoracic sections were cut open longitudinally along the inner curvature. Outer curvatures were subsequently cut longitudinally from the proximal ascending to the subclavian arterial branch (A). Aortas were pinned onto a wax dish and intimal area of the ascending aortas was measured between the yellow lines (B).(PDF)Click here for additional data file.S8 FigNumbers below images are ascending aortic area measurements.(PDF)Click here for additional data file.S9 FigNumbers below images are suprarenal aortic diameter measurements.(PDF)Click here for additional data file.S10 FigNumbers below images are ascending aortic area measurements.(PDF)Click here for additional data file.S11 FigNumbers below images are suprarenal aortic diameter measurements.(PDF)Click here for additional data file.S12 FigTeal lines indicate the outer curvature and diameter measurements of ascending aortas.(PDF)Click here for additional data file.S13 FigTeal lines indicate the outer curvature and diameter measurements of ascending aortas.(PDF)Click here for additional data file.S14 FigNumbers below images are ascending aortic area measurements.(PDF)Click here for additional data file.S15 FigNumbers below images are suprarenal aortic diameter measurements.(PDF)Click here for additional data file.S16 FigMice were infused with AngII for 28 days then injected with mouse TGF-\u03b2 neutralizing IgG and infused with AngII for an additional 28 days. Numbers below images are ascending aortic area measurements.(PDF)Click here for additional data file.S17 FigMice were infused with AngII for 28 days then injected with mouse TGF-\u03b2 neutralizing IgG and infused with AngII for an additional 28 days. Numbers below images are suprarenal aortic diameter measurements.(PDF)Click here for additional data file."} +{"text": "Multidrug-resistant and lately pan drug-resistant (PDR) Acinetobacter Baumanii presents an increasing challenge to health care and especially in Intensive Care Units (ICU).To evaluate the efficacy of daptomycin and vancomycin against PDR Acinetobacter Baumanii blood stream infections in ICU. In vitro, the administration of antibiotics usually used against Gram positive cocci (like vancomycin or daptomycin) has resulted in good bacterial response against PDR Acinetobacter even though it's a Gram - bacterium. However, clinical data to support the use of these regimens are still lacking.From June 2014 to March 2015 all PDR Acinetobacter Baumanii blood stream infections along with the given therapeutic regimens were recorded. Age, sex, total days of antibiotic administration and culture results were recorded.4 blood stream infections in 4 patients were recorded for the time interval mentioned. Among those, 3 were originally receiving intravenous colistin plus meropenem while one was only taking meropenem. As soon as PDR blood stream infection was confirmed daptomycin 6mg/kg (average 9 days of administration) was started on 2 patients while vancomycin 15mg/kg/12 hours (average 8 days of administration)was added to the other two cases. Subsequent blood cultures came back negative after 2-3 days (2.2 days) of treatment.Despite the small number of patients daptomycin and vancomycin seems to be effective against PDR Acinetobacter Baumanii blood stream infections. Further studies are needed to support the proposed scheme."} +{"text": "Cardiac MRI has been shown to be the gold standard for noninvasive assessment of myocardial fibrosis. The extent of fibrosis has been associated with worse outcomes in patients undergoing CABG surgery. The presence of fibrosis and associated clinical features of patients with chronic mitral regurgitation undergoing mitral valve repair have not been described. The objective of this study is to examine prognostic value of preoperative myocardial fibrosis assessment by cardiac MRI in patients with chronic mitral regurgitation undergoing mitral valve repair.This is a prospectively enrolled consecutive series of 48 subjects with chronic mitral regurgitation who underwent cardiac MRI followed by mitral valve repair using the non-resectional dynamic technique from August 2008 - April 2013. Pre-operative fibrosis burden, other cardiac MRI and clinical variables were collected. Differences among variables between subjects with and without fibrosis were statistically calculated.The cohort had a mean (SD) age of 61(13) years and included 33 men (69%). Median postoperative follow up interval was 53.5 days (Interquartile range 10 and 442 days). Mean (SD) interval between cardiac MRI and mitral valve repair was 19.58 (18.37) days. Fibrosis was present in 19 subjects (39.6%) with a median fibrosis burden of 4% of the left ventricle. Within the fibrosis group, 9 (47.4%) had a non-coronary artery disease fibrosis pattern. Comparison of selected baseline clinical and cardiac MRI variables is shown in the Table Presence of preoperative myocardial fibrosis derived from cardiac MRI is associated with worse postoperative outcomes in patients with chronic mitral regurgitation undergoing mitral valve repair.None."} +{"text": "Presence/absence variation (PAV) of genes was also detected. We found that substantial sequence variation exists among genomic regions targeted in this study, which was particularly evident within coding regions. This diversification has the potential to broaden functional diversity and generate phenotypic variation that may lead to new adaptations and the modification of important agronomic traits. Further, annotated SNPs identified here will serve as useful genetic tools and as candidates in searches for phenotype-altering DNA variation. In summary, we demonstrated that sequencing of captured DNA is a powerful approach for variant discovery in maize genes.A major goal of maize genomic research is to identify sequence polymorphisms responsible for phenotypic variation in traits of economic importance. Large-scale detection of sequence variation is critical for linking genes, or genomic regions, to phenotypes. However, due to its size and complexity, it remains expensive to generate whole genome sequences of sufficient coverage for divergent maize lines, even with access to next generation sequencing (NGS) technology. Because methods involving reduction of genome complexity, such as genotyping-by-sequencing (GBS), assess only a limited fraction of sequence variation, targeted sequencing of selected genomic loci offers an attractive alternative. We therefore designed a sequence capture assay to target 29 Mb genomic regions and surveyed a total of 4,648 genes possibly affecting biomass production in 21 diverse inbred maize lines . Captured and enriched genomic DNA was sequenced using the 454 NGS platform to 19.6-fold average depth coverage, and a broad evaluation of read alignment and variant calling methods was performed to select optimal procedures for variant discovery. Sequence alignment with the B73 reference and Zea mays subsp. mays, commonly referred to as maize, reference genome sequence . Details are provided in The raw 454 DNA sequence data obtained from sequence capture and re-sequencing of the 21 maize inbred lines have been deposited at the European Nucleotide Archive (ENA) in the Sequence Read Archive (SRA) and are available under the following EBI project ID: PRJEB5496, [S1 FigFor each particular setting, the mapping is evaluated for all 21 inbred lines by determining the number of aligned reads, the number of aligned read on target, the mutual agreement of the alignment (\u2018number of reads mapped by other method\u2019), the uniquely aligned reads, and the number of reads aligned by the majority of methods. The graphic shows the results for genotype NC358, which was selected as example.(TIF)Click here for additional data file.S2 FigThe analysis was performed on the random selected genotype \u2018NC358\u2019, and all 504 possible combinations of alignment and variant calling methods (three parameter settings) were included. To construct the heat map, the detected VPs of each individual approach were compared to the 50k data to reveal true positive predictions.(TIF)Click here for additional data file.S3 Fig1-score, shown in yellow, illustrates the impact of false positive and false negative values. The harmonic mean reaches highest values at VCC3.All VPs in our studied lines that overlap a 50k position are considered true positive. The proportion of the total number of true positives is depicted in blue for each VCC value. The F(TIF)Click here for additional data file.S4 Fig(TIF)Click here for additional data file.S5 FigChromosome bins with a size of 2 Mbp were analyzed, and regions that are characterized with high diversity in our genotype collection are indicated in yellow.(TIF)Click here for additional data file.S6 FigChromosomes were analyzed in 500 kbp bins, and the genotypes are shown in the identical order as depicted in (TIF)Click here for additional data file.S7 Fig(JPG)Click here for additional data file.S8 FigThe Blast2GO hierarchy is presented at level three for all three categories.(TIF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 TableThe number of non-synonymous coding SNPs and nonsense SNPs, and the number of genes they affect is also shown.(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S7 Table(XLS)Click here for additional data file.S8 Table(XLSX)Click here for additional data file.S9 TableThe resulting sequence coverage is presented as percentage of gene length.(XLSX)Click here for additional data file.S10 TableA joint analysis links the phenotypic information of lowest yielding (B111 and EA1070) and highest yielding (F2 and F7) inbred lines with the presence and absence information. A gene was declared absent if at least 75% of sequence length is missing, and the captured gene length of maize inbred B73 is used as reference.(XLSX)Click here for additional data file.S11 Table(XLS)Click here for additional data file.S12 Table(XLS)Click here for additional data file.S13 Table(XLSX)Click here for additional data file.S14 Table(XLS)Click here for additional data file.S15 Table(XLSX)Click here for additional data file.S16 Table(XLSX)Click here for additional data file.S17 Table(XLSX)Click here for additional data file.S18 TableThe analysis included 4,342 non-overlapping genes that were mapped uniquely to the maize genome reference (v3). Results revealed different representations of sequence capture probes in UTR, exonic and intronic regions of studied genes.(XLSX)Click here for additional data file.S1 File(TXT)Click here for additional data file.S2 File(TXT)Click here for additional data file."} +{"text": "MYB activation is proposed to underlie development of adenoid cystic cancer (ACC), an aggressive salivary gland tumor with no effective systemic treatments. To discover druggable targets for ACC, we performed global mRNA/miRNA analyses of 12 ACC with matched normal tissues, and compared these data with 14 mucoepidermoid carcinomas (MEC) and 11 salivary adenocarcinomas (ADC). We detected a unique ACC gene signature of 1160 mRNAs and 22 miRNAs. MYB was the top-scoring gene (18-fold induction), however we observed the same signature in ACC without detectable MYB gene rearrangements. We also found 4 ACC tumors (1 among our 12 cases and 3 from public databases) with negligible MYB expression that retained the same ACC mRNA signature including over-expression of extracellular matrix (ECM) genes. Integration of this signature with somatic mutational analyses suggests that NOTCH1 and RUNX1 participate with MYB to activate ECM elements including the VCAN/HAPLN1 complex. We observed that forced MYB-NFIB expression in human salivary gland cells alters cell morphology and cell adhesion in vitro and depletion of VCAN blocked tumor cell growth of a short-term ACC tumor culture. In summary, we identified a unique ACC signature with parallel MYB-dependent and independent biomarkers and identified VCAN/HAPLN1 complexes as a potential target. MYB gene were detected in a subset of human T cell acute lymphoblastic leukemia (T-ALL) [in vitro transforming activity that is largely restricted to hematopoietic animal model systems. Therefore, the identification of a recurrent MYB:NFIB translocation in ACC offered a new opportunity to study MYB biology in a defined epithelial human cancer model system.Malignant epithelial salivary gland tumors (SGTs) represent a heterogeneous group of tumors, including the major subtypes of adenoid cystic carcinomas (ACC), mucoepidermoid carcinomas (MEC), and adenocarcinomas (ADC) . Each of (T-ALL) and over (T-ALL) -24. EctoThe terminal NFIB exon, containing a small conserved open reading frame, is a recurrent translocation 3\u2032 gene fusion partner in salivary gland ACC , pleomorWhile MYB rearrangements were detected by FISH in approximately 50% of cases, there was an overall low rate of somatic mutations suggesting candidate cooperating driver mutations -32. ThesTo address these uncertainties, we have now performed global mRNA and miRNA expression analysis for a large collection of well-annotated malignant SGT and their matched normal tissues and integrated this data with somatic mutational analyses. We also performed functional assays to determine the transforming activity of activated MYB by either 3\u2032 truncation or by fusion with terminal NFIB exon(s). We have detected a distinct ACC mRNA signature that includes over-expression of a network of extracellular matrix (ECM) components that arise from both MYB-dependent and -independent signals. These data suggests a cooperating role between MYB and RUNX1 /NOTCH1 signaling and proposes new therapeutic targets for this lethal disease.To define a diagnostic adenoid cystic cancer (ACC) gene signature and to identify new therapeutic targets, we performed global mRNA and miRNA expression array analysis for 12 ACC tumors and matched adjacent normal samples. We also compared the ACC signatures to 14 mucoepidermoid cancers (MEC) and 11 salivary adenocarcinomas (ADC). All samples showed high RNA integrity (RIN >8) and passed quality controls described in Methods. We first performed principle component analysis Figure , both unAn unbiased inspection of global ACC gene expression identified MYB as the top differentially expressed gene as compared to either matched normal tissue or to other SGT subtypes such as MEC and ADC Figure . We confThe detection of a similar ACC mRNA gene signature in both MYB fusion-positive and fusion-negative tumors as well as a sample with low MYB expression suggested several alternative mechanisms: i) presence of subtle variant MYB fusion events that were not detected in our samples with RT-PCR. ii) alternative, non-fusion somatic events that activate MYB, or iii) activation of an alternative gene pathway that mimics activated MYB signaling. To address the first possibility, we re-analyzed the relative expression levels of MYB exons using detailed exon array expression data. This microarray platform allows for multiple probes covering most MYB exons and we are able to generate \u2018exon-plot\u2019 images to examine exon-specific expression except for MYB exon 10 that was not present in ACC tumors. As expected, we were able to infer the approximate MYB breakpoints by noting relative drops in average probe set intensities immediately after the breakpoints in validated MYB-NFIB fusion positive samples. In contrast, true fusion-negative samples showed consistent probe intensities across the whole gene. With this \u2018exon-plot\u2019 approach, we identified two misdiagnosed \u2018fusion negative\u2019 tumors Figure , asterixAs noted in Figure HAPLN1 expression [We then integrated our new ACC gene signature with data from three recent whole exome ACC sequencing studies -32 and ipression , 42. Thehsa-miR-150 was down-regulated in all ACC tumors ; MYB with only 5\u2032UT (MYB no 3\u2032UTR), truncated MYB with a stop codon inserted at the most common translocation breakpoint in exon 15 (Trunc MYB), and MYB-NFIB fusion (MYB-NFIB) Figure . We obtag Figure . A similg Figure . These do Figure and alsoo Figure with botACC is one of the most common malignant SGT subtypes with no curative strategy for unresectable disease. In addition, lethal metastatic recurrences can present many years after the initial surgical resection that has further stimulated an urgent search for new therapies. Cytotoxic chemotherapy has not been effective for advanced disease and clinical trials testing imatinib to target KIT over-expression or lapatIn fact, we discovered that 30% of the top ranking genes measured by fold overexpression were localized within the extracellular space and this pattern appeared unique for ACC as compared to other SGT subtypes such as mucoepidermoid carcinomas and adenocarcinomas. Unexpectedly, we also detected an ACC sample with low MYB expression that still retained this global ACC gene expression signature. We validated this observation by identifying 3 additional ACC tumor samples from the public microarray databases with low MYB levels suggesting that approximately 5-10% of ACC samples may have this alternate phenotype with negligible MYB levels. Although MYB activation defines ACC tumorigenesis and MYB overexpression has been associated with a worse clinical outcome in patients with ACC , 47-49, in vitro and in vivo with short term ACC cell cultures following depletion of VCAN and the generation of new validated immortalized ACC tumor cell lines and animal models will allow preclinical testing of these new promising extracellular targets for development of new treatment strategies.To pursue this possibility we performed an integrated analysis of gene expression and somatic mutational data and these data proposed a role for MYB, NOTCH1, and RUNX1 in regulating a subset of key ACC target genes. RUNX1, a MYB transcriptional co-factor, may be a specific target for missense or C-terminal truncated activating mutations in ACC that cooperate to deregulate MYB transcriptional activation -55. For A total of 74 salivary gland tumors including 24 salivary gland adenoid cystic cancer, 28 mucoepiderrmoid cancer and 22 adenocarcinoma samples were excised from human patients under approved IRB protocols from University of Texas MD Anderson Cancer Center. MYB-NFIB fusion positive tumors were preselected by RT-PCR. Total RNAs were extracted and evaluated using NanodropTM 8000 and the Agilent 2100 Bioanalyzer with RNA 6000 Nano Labchip\u00ae .\u00ae_human gene 1.0_ST and miRNA_GeneChip_1.0; Affymetrix, CA). Data were processed through the robust multichip average algorithm and quantile normalization methods followed by log2 transformation [\u00aeGenomics . Both baseline filter (intensity >=50) and variation filter (|fold-change|>=2) were applied before mRNA differential expression analysis. Both baseline filter (intensity >=800) and variation filter (|fold-change|>=2) were applied for microRNA chip analysis. Hierarchical cluster analysis and the principle component analysis were performed for all samples using SAS . Paired t-tests were performed to compare tumor samples to their matched normal tissues and Hochberg-Benjamini FDR was set at 0.05 [Microarray assays were performed by following manufacturer's instructions and inserted into the pAcGFP1N1 expression plasmid using NotI and KpnI restriction sites and the terminal NFIB exon with or without NFIB 3\u2032UT sequences was introduced by PCR-mediated cloning. All constructs were validated by sequencing. For stable clone selection, 50 ng/ml Geneticin\u00ae selective antibiotics was introduced after 48 hours of transfection. The pLKO.1 lentiviral VCAN-shRNA constructs were obtained from Open Biosystems . The lentiviral procedures applied for shRNAi were performed as described in [ribed in . The pBA\u00ae). HEK293T cells were cultured in high glucose DMEM medium supplemented with 10% FBS and 1% pen/strep (Gibco\u00ae). RK3E cells were cultured in RPMI 1640 (Sigma) with 10% FBS and 1% pen/strep (Gibco\u00ae). Normal human immortalized salivary gland cells [\u00ae, Cat17005-075). Defined Trypsin Inhibitor was also purchased from Invitrogen (CatR-007). The primary normal salivary gland cell culture for wt FVB mice was performed following procedures described in [NIH/3T3 cells were cultured in DMEM (Sigma) supplemented with 10% FBS and 1% pen/strep and TaqMan Fast Universal PCR Master Mix . All gene expression probes were commercially purchased (Applied Biosystems\u00ae). Relative expression of target gene was calculated in comparison to 18s rRNA values. The quantitative miRNA RT-PCR analysis was performed by following procedures as described in [Reverse transcription and amplification of cDNA was performed in the 7900 HT Fast (Applied Biosystems) system by following manufacture instruction using High-Capacity cDNA Reverse Transcription Kits (Applied Biosystemsribed in .Cells pellet was lysed in RIPA buffer (Boston Bioproducts) and subjected to SDS-polyacrylamide tris-glycine gel separation (Invitrogen). And the membrane transfer was performed using iBlot\u00ae 7 Minute Blotting System (Invitrogen) following the manufacturer's instructions. Membrane was blocked with 3% BSA overnight at 4\u00b0C and then incubated with primary antibodies for 1 hour at room temperature. Antibody against N-terminus MYB was purchased from Abcam (EP769Y). After washed with PBS, the membrane was incubated with a horseradish peroxidase-conjugated secondary antibody for 45 minutes. Bands were visualized with Chemiluminescence (Pierce).The protein half life was determined by cycloheximide blocking assay. Cells were seeded into 100mm dish and subjected to inhibition of protein synthesis with 100 \u03bcg/ml cycloheximide (Sigma) for 10 minutes, 20 minutes, 30 minutes, 60 minutes, 120 minutes or no treatment as control. Cycloheximide treated cells were then harvested and subjected to SDS-PAGE gel separation and western-blot analysis for retained protein. The relative protein quantity were measured using ImageJ .5 trypsinized cells were subjected to soft-agar assay as described in [2 for 3 weeks. And cells were feed once every week. Colonies were counted manually under10x microscope .A total of 1\u00d710ribed in . General6 cells were seeded into 6-well plates in day one, and then viable cells in the culture supernatant were counted with TrypanBlue (Gibco) staining at day 4. Unstained detached cells were washed twice with PBS, reseeded to 6-well plates for growth and then stained with crystal violet for cell density observation after 4 days.A total of 1\u00d7106 cells were washed twice with PBS and then subjected to subcutaneous flank injection into NOD.SCID mice . Freshly prepared human ACC tumor obtained under IRB-approved tissue protocol was also subjected to xenograft tumor growth and short term cell culture. Xenograft tumor volumes were calculated as: volume = (width)2 \u00d7 length/2. The primary tumor cell culture was performed using methods as described in [A total of 1\u00d710ribed in , 76. The"} +{"text": "Cellular levels of cyclic GMP (cGMP) are tightly controlled by synthetic and degradative mechanisms. Pharmacological manipulation of these processes augments cGMP-dependent signalling and is beneficial in treating cardiovascular disease (e.g. pulmonary hypertension). An additional mechanism potentially important in regulating cGMP signalling is cellular extrusion, driven by a family of multidrug resistance proteins (MRPs). Herein, we investigated if inhibition of MRPs modulates vascular reactivity, smooth muscle cell proliferation, and systemic hemodynamics.The functional reactivity of murine aortic rings and proliferation of human pulmonary artery smooth muscle cells (PASMC) were determined in the absence and presence of the MRP inhibitor MK571. Hemodynamic changes in vivo in response to MK571 were analysed acutely by bolus dosing and chronically by radiotelemetry.In vivo, MK571 elicited an acute, dose-dependent hypotensive activity and when delivered via the drinking water caused a more sustained drop in mean arterial pressure (~5 mmHg).MK571 (1nM-50\u00b5M) caused a concentration-dependent relaxation of mouse aortic rings. In the presence of a threshold concentration of MK571 (3\u00b5M), vasorelaxant responses to NO and atrial natriuretic peptide (ANP) were significantly augmented. MK571 (3\u00b5M) also significantly inhibited PASMC proliferation and enhanced the anti-mitogenic properties of ANP (1\u00b5M) and NO. These data suggest that extrusion by MRPs contributes to the dynamic equilibrium regulating intracellular levels of cGMP, and may represent a further target amenable to drug intervention for the treatment of cardiovascular disease."} +{"text": "Autosomal dominant polycystic kidney disease (ADPKD) is a systemic conditions with cardinal symptom of bilateral multiple renal cysts of variable sizes. ADPKD is genetically heterogeneous, Mendelian disorder, shows late onset and contributes 8-10% cases of end stage renal disease (ESRD). Linkage analysis in ADPKD familial cases revealed approximately 85% cases of PKD1, rest cases of PKD2 and an unidentified locus.In the present study occurrence of mutation, amino acid change and three frequently observed SNPs from 3\u2019single copy region (exon 44-46) and in 5\u2019 duplicated region (exon 2-11) of were screened in 47 patients and in control individuals to search the responsible and/or susceptible spots for ADPKD in Indian patients using direct DNA sequencing of specified exons and RFLP for SNP assay. PKD1 exons 44-46 encoding intracellular and exons 2-11 encoding ligand binding domains, Ig-like PKD domains in extracellular compartment with their suggested role in cell signaling and cell adhesion were chosen for this study.Screening of exons 44-46 in 47 patients identified five exonic and two intronic variants. Assessment of the three SNPs viz. I4045V, A4059V, and A4092A screened in a multiplex family with four affected and 30 unaffected members along with 100 ages and sex matched controls showed higher occurrence (2X) in patients compared to controls indicating their role for disease susceptibility. Screening of exons 2-7 in 19 patients uncovered six exonic and two intronic variants while screening of exons 8-11 in 15 patients detected twelve exonic and one intronic (IVS9+14_27del14) variants. Two exonic changes generate 288 amino acids long truncated protein and a stretch of nine altered amino acids sequence in LDL-A domain respectively.The spectrum of changes detected in various affected individuals indicates that the clinical as well as allelic heterogeneity compounding the pathophysiology of ADPKD in both sporadic and familial cases. Screening of remaining region of PKD1 and other candidate genes is therefore planned for future for better understanding."} +{"text": "Post-translational modification of the neural cell adhesion molecule (NCAM) by polysialic acid (PSA) is crucial for nervous system development and brain plasticity. PSA attachment is catalyzed by the two polysialyltransferases, ST8SiaII and ST8SiaIV. ST8SiaII dominates during embryonic and early postnatal development while ST8SiaIV is the prevailing enzyme of the juvenile and adult brain. A growing body of evidence links aberrant levels of NCAM and PSA to neuropsychiatric disorders, including schizophrenia. To investigate whether polysialyltransferase deficiency might cause a schizophrenia-like phenotype, ST8SiaII-/- mice, ST8SiaIV-/- mice and their wild-type littermates were assessed neuroanatomically and subjected to tests of cognition and sensory functions. ST8SiaII-/- but not ST8SiaIV-/- mice displayed enlarged lateral ventricles and a size reduction of the thalamus accompanied by a smaller internal capsule and a highly disorganized pattern of thalamocortical and corticothalamic fibers. Loss of ST8SiaII was associated with reduced phosphorylation of fibroblast growth factor receptor 1 in the frontal cortex of newborn mice and retarded neuronal differentiation of newly generated cells in the dentate gyrus of adults. ST8SiaII-/- and ST8SiaIV-/- mice were both impaired in short- and long-term recognition memory, but only ST8SiaII-/-mice displayed impaired working memory and a deficit in prepulse inhibition, which could be attenuated by clozapine treatment. Furthermore, ST8SiaII-/- mice exhibited anhedonic behavior and increased sensitivity to amphetamine-induced hyperlocomotion. These data reveal that reduced polysialylation in ST8SiaII-/- mice leads to pathological brain development and schizophrenia-like behavior. We therefore propose that ST8SiaII deficiency has the potential to cause a neurodevelopmental predisposition to schizophrenia."} +{"text": "The advent of combined antiretroviral therapy (ART) in the past decade has led to HIV suppression in most cases. Virological failure was the main reason for ART switch a few years ago; however, toxicity and treatment simplification have now gained importance due to the availability of more effective and convenient drugs. This study assessed the reasons for ART switch in daily practice.Observational retrospective study that included patients whose ART was switched between January 2011 and July 2012. Patients with any other switch during the follow-up period (until September 2013) were excluded.A total of 246 patients were included. Main reasons for ART switch were simplification (33%) and toxicity (31%), followed by clinical trial inclusion (13%), virological failure (6%), drug interaction (4%), patient decision (3%), lack of adherence (2%), pregnancy (1%) and other (8%).Eighty patients switched to a simpler regimen . In this case, previous ART mostly included 2NRTI+1PI/r (54%) . The simThe main reasons for ART switch in daily practice are simplification and toxicities, renal and CNS toxicities being the most prevalent. The preferred simplification strategies are nuke-sparing regimens, mainly DRV/r-based monotherapy and dual therapy. ART switch leads to a complete resolution of toxicities in most cases in the short term."} +{"text": "The Least Square Fitting (LSF) method is a statistical approach used for evaluating respiratory mechanics . It alloWe compared the LSF performance during PSV and NAVA. Our hypothesis was that the LSF works better during NAVA than during PSV, since NAVA algorithm allows a more accurate neuro-ventilatory coupling.T/Crs + PEEPtot during inspiration. The coefficient of determination (CD) of the applied equation was used to compare data obtained during NAVA and PSV, the higher being the CD, the better the quality of the data. These data were obtained at the beginning of mechanical ventilation (T0), and after 12 (T12), 24 (T24), 36 (T36), 48 (T48), 60 (T60) and 72 (T72) hours of mechanical ventilation.15 patients undergoing mechanical ventilation for acute respiratory failure were ventilated using randomly either PSV or NAVA. Data of resistance (Rrs), elastance (Ers) and total positive end expiratory pressure (PEEPtot) were obtained by fitting the equation Paw = Rrs x V\u00b4 + VData obtained with LSF were statistically more reliable during NAVA than during PSV (Chi-squared test: p < 0.001). The CD level showed a higher value during NAVA (T0 median 0.9855), that was maintained constantly higher in time, than during PSV (T0 median 0.9288), in which the value of the CD progressively worsened by the hours of mechanical ventilation.The LSF method of the LSF performs better during NAVA then during PSV. By the hours of mechanical ventilation the performance of the LSF method further worsens during PSV while remains constant during NAVA. Our data indirectly confirm more physiological patient-ventilation interactions during NAVA than during PSV."} +{"text": "Objectives: Estrogen deficit following menopause results in cognitive behaviors impairment. This study aimed to evaluate the effects of pomegranate seed extract (PGSE) on avoidance memories after permanent bilateral common carotid arteries occlusion (2CCAO) in ovariectomized (OVX) rats.Materials and Methods: Adult female Wistar rats were divided randomly into eight groups with 8 rats in each group: 1) Sham-operated for ovaries and 2CCAO (ShO); 2) OVX and sham operated for ischemia (OShI); 3-7) OVX with 2CCAO (OI) received PGSE for 14 days ; 8) OShI received most effective dose of PGSE (200 and 400 mg/kg for passive and active avoidance memories respectively). Active and passive avoidance tasks were measured in Y-maze and two-way shuttle box respectively. Data were analyzed with one-way and RM-ANOVA followed by HSD post-hoc test.Results: Sensorimotor impaired in OShI+Veh and OI+Veh (P<0.001 vs. ShO). PGSE improved it significantly in dose dependently manner (P<0.001 vs. OI+Veh). Both types of memories were significantly impaired in OVX rats before and after 2CCAO (P<0.001). PGSE treatment significantly improved memories in OI groups compared with OI+Veh. No toxicity was observed with PGSE consumption . Conclusion: PGSE exhibits therapeutic potential for avoidance memories, which is most likely related at least in part to its phytoestrogenic and also antioxidative actions. The effects of combined gonadal hormones deprivation and cerebral hypoperfusion/ischemia on memory is unknown. Therefore, we hypothesized that phytoestrogenic natural substances such as pomegranate seed extract (PGSE) serves as a neurotrophomodulatory substance for some brain areas neurons thought to be involved in learning and memory. Gonadal hormones, such as estrogen, can alter cognitive performance. Estrogen can have positive mnemonic effects in the inhibitory avoidance task can be effective for the improvement of post-ischemic injuries. On the other hand, female sex hormones such as estrogen are neuroprotective and PGSE also contains phytoestrogenic substances with antioxidative properties, we decided therefore to test the effect of PGSE on cognition deficiency induced by HI in adult female rats. The present study investigated the effects of different doses of PGSE on passive and active avoidance memories following cerebral hypoxia-ischemia induced via bilateral common carotid artery occlusion. Animalsad libitum except during the experiments. Animal handling and experimental procedures performed under observance of the University and Institutional legislation, controlled by the Local Ethics Committee for the Purpose of Control and Supervision of Experiments on Laboratory Animals. All efforts were made to minimize animal suffering, to reduce the number of animals used. Prior to the onset of behavioral testing, all rats were gentle handled for 3 days (daily 10 min). After one week, animals of seven groups were ovariectomized (O) under anesthesia induced by injection of ketamine hydrochloride and Xylazine . While ovaries remained intact in sham operated groups (ShO). Bilateral common carotid arteries were occluded and dissected (2CCAO) in ovariectomized rats (OI groups). Animals were divided randomly into eight groups with 8 in each: 1) sham-operated for OVX (ShO); 2) ovariectomized and sham operated for ischemia with manipulation of both common carotids arteries without occlusion received vehicle (OShI+Veh); 3-7) ovariectomized with 2CCAO (OI) received PGSE for 14 days ; 8) OShI received most effective dose of PGSE which was different in two cognition tests . Eighty adult male albino rats of Wistar strain obtained from Ahvaz Jundishapur University of Medical Sciences (AJUMS) laboratory animal center were used in this study. Animals were housed in standard cages under controlled room temperature (20\u00b12 \u25e6C), humidity (55-60%) and light exposure conditions 12:12 h light\u2013dark cycle (lighted on 07:00 am). All experiments carried out during the light phase of the cycle (8:00 am to 6:00 pm). Access to food and water were 2CCAO procedure Cechetti\u2019s method (2012) with little modification was used. Briefly, rats were anesthetized with ketamine/xylazine . A neck ventral midline incision was made and the common carotid arteries were then exposed and gently separated from the vagus nerve. Carotids were occluded with three days interval between interventions, the right common carotid being the first to be assessed and the left one being occluded three days later. Sham-operated rats were under same surgical procedures without carotid artery ligation and occlusion as large fruit with red barriers were purchased from Shahreza granatum gardens- Iran. Seeds removed from the fruits, air dried in shade for one week and milled to fine powder . The seeds powder was macerated in 70% ethanol for 72 hours at room temperature. The ethanol extract evaporated to remove ethanol and PGSE was obtained as a lyophilized powder (yield 17\u00b12%). Pomegranate fruits everyday 8:00\u20139:00 am for 2 weeks, starting on the 5 days post ischemic injury. Sham treated animals (OI+Veh) were received same volume of normal saline (2ml) for same period. Sensorimotor evaluation It consisted of two tests developed and described by Garcia rat moves around, explores the environment, and approaches at least three walls of the cage; (2) rat moves around in the cage but does not approach all sides and hesitates to move, although it eventually reaches at least one upper rim of the cage (height=10 cm); (3) rat dose not rise up at all and barely moves in the cage; (4) rat does not move at all. Symmetry in the movement of four limbsThe rat is held in the air by the tail to observe symmetry in the movement of the four limbs. Scores indicate the following: (1) all four limbs extended symmetrically; (2) limbs on one side extended less or more slowly than those on the other side; or slow extension of the four limbs; (3) limbs on one or both sides show minimal movements; (4) forelimbs on one or both sides do not move at all.Passive avoidance task The apparatus used for evaluation the passive avoidance task was two-way shuttle box , which consisted of two adjacent plexiglas compartments of identical dimensions (27\u00d714.5\u00d714 cm) with grid floors. The floor of two compartments has been covered with stainless steel bars (2 mm diameter) with 1 cm distance. Light compartment was illuminated by a 5 W lamp mounted on its wall just below a movable transparent plexiglas ceiling. The Tamburella\u2019s procedure with little modification was used for passive avoidance memory test . The Yu and Besnard procedures with little modifications were used for active avoidance memory test and repeated measures (RM) ANOVA (for active avoidance task) followed by HSD post hoc test. A P-value less than 0.05 were assumed to denote a significant difference and levels of significance are indicated by symbols: * vs. ShO group and # vs. OShI+Veh or OI+Veh groups. Motor evaluationTreatment with different doses of PGSE for two weeks improved sensorimotor scores significantly in OI+E200, OI+E400 and OI+E 800 groups, but not with dose 100 mg/kg (P<0.001 vs. OI+Veh), while there was not any difference between them. There were significant differences between OShI+Veh vs. ShO (P<0.001) and OShI+E200 vs. OShI+Veh groups respectively.Passive avoidance memoryAs shown in Treatment OVX animals without ischemia with this effective dose of PGSE (OShI+E200) compensated it and reversed its level in ShO group . TreatmeAs shown in Active avoidance memoryResults of treatment the ovariectomized with 2CCAO groups with different doses of PGSE showed that only doses 400 and 800 mg/kg could improve %CCRs during 2-4 sessions significantly (P<0.001), with no significant difference. But it is steel lower than ShO group significantly during those training sessions and 400 mg/kg (for active avoidance task) had significant improving effects. These results show that 2CCAO induction causes oxidative stress progression and free radicals production in brain tissues and PGSE as an estrogenic compound and a potent antioxidant could activate estrogen beta receptors and beta (ERbeta) in the adult brain modulating memory, learning, mood and acts as a neuroprotector. As estrogen levels change with age, especially in females, it is important to know the effects of low E2 levels on ERalpha distribution; results from previous studies are controversial regarding this issue and reperfusion have been developed in recent years, but none has been highly successful. A fundamental process believed to be responsible for HI damage to neurons is excitotoxicity, triggered mainly by elevated extracellular glutamate concentration showed that middle cerebral artery occlusion (MCAO) causes a larger cortical-striatal infarct in this older acyclic group compared with younger females. Moreover, although estrogen treatment is neuroprotective in younger females, estrogen paradoxically increases infarct volume in acyclic females. These data support the hypothesis that stroke severity in older females is associated with decreased IGF-1 and further indicate that short-term post-ischemic IGF-1 therapy may be beneficial for stroke (Selvamani & Sohrabji, 2010The results of present study shows the PGSE is a natural product with power phytoestrogenic and antioxidative effects without any side effects could improve the cognitive deficit during menopause period and specially as followed by stress oxidative and possible cerebral hypoperfusion induced by vascular obstruction. Our results show that passive and active avoidance tasks impair after ovarectomy in rats because of removing peripheral source of estrogen and treated them with PGSE as a natural product with phytoestrogenic and antioxidant contents could reverse both active and passive avoidance tasks significantly in OVX rats suffer with cerebral HI. Our previous results showed that 2CCAO induction causes oxidative stress progression and free radicals production in brain tissues (Mansouri et al., 2013"} +{"text": "Only a minority of patients suffering out-of-hospital cardiac arrest receive any bystander cardiopulmonary resuscitation (CPR). Bystander CPR is associated with improved odds for survival. The PulsePoint smartphone application alerts users in the vicinity of a cardiac arrest to facilitate immediate citizen bystander resuscitation. Addressing implementation barriers may provide an opportunity to increase effectiveness of the application. PulsePoint is currently active in over 400 communities in the United States. Our objective was to identify modifiable barriers to optimal implementation of the PulsePoint smartphone application.We conducted a structured survey delivered via Survey Monkey optimized for mobile devices. All alerted PulsePoint users between 28 June 2012 and 1 November 2013 were sent an invitation to participate in the survey. Survey responses associated with test activations and emergency medical services (EMS) professionals on active duty were excluded from the analysis.Of 4,827 survey requests sent, 995 responded and completed our survey (response rate 21%). Respondents identified themselves as firefighters (30%), paramedics (19%), EMTs (16%), nurses (4.4%), MDs (0.83%) and other professions (50%). Of those who received a PulsePoint alert, 23% (157/690) responded by making their way towards the location of the emergency. Of those who responded, 70% (110/157) arrived on scene. Most of those who did not arrive said they saw professional rescuers already on scene and turned back . Of those who arrived on scene, only 34% (37/110) found a person unconscious and not breathing normally. The majority found a person on scene who was not suffering cardiac arrest. Of those who arrived on scene prior to EMS and also found a cardiac arrest victim on scene who required resuscitation, 80% (8/10) performed bystander CPR.We observed a very high proportion of bystander CPR (80%) for victims of out-of-hospital cardiac arrest when PulsePoint users arrived before EMS. This suggests that optimized PulsePoint implementation may increase community bystander CPR rates. Alert specificity for cardiac arrest may be too low (the cry wolf scenario) with current default activation criteria. Also, the current activation radius (0.5 mile) may be too large because EMS frequently arrive before the PulsePoint responder."} +{"text": "There was no personal / family autoimmunity history. A paternal grandparent was thought to have type 2 diabetes. She was haemodynamically stable and mildly dehydrated without clinical signs of insulin resistance. She had bilateral stellate posterior subcapsular cataracts. Plasma glucose was 57mmol/L with normal blood ketones and venous pH. HbA1c was 18.2% (175mmol/mol). Mildly elevated urea and creatinine corrected after rehydration. Liver function tests, serum magnesium and urate were normal. Islet autoantibodies were negative.A 13 year old pubertal Caucasian female presented with longstanding polyuria/polydipsia and weight loss of 12kg over 4 months (BMI decreased from 23 to 17.5kg/mth centile), with echogenic parenchyma, reduced corticomedullary differentiation, normal cortical thickness and a 5mm left cortical cyst. Urinary albumin/creatinine and calcium/creatinine were normal.Renal ultrasound (US) identified bilaterally small kidneys (<5Further imaging (US and MRI) demonstrated severe pancreatic atrophy and a m\u00fcllerian duct anomaly (arcuate / subseptate uterus).Faecal elastase 1 in a soft stool sample was low (range of severe exocrine pancreatic insufficiency). However the patient has gained weight with insulin treatment and had normal measures of fat-soluble vitamins.Conduct genetic testing for the autosomal dominant Renal Cysts and Diabetes Syndrome (RCAD), also known as MODY5, caused by heterozygous mutations of the hepatocyte nuclear factor\u20131 beta (HNF1B) gene (chromosome 17q12).Direct testing for HNF1B gene sequence variations was performed by Multiplex Ligation-dependent Probe Amplification (MLPA) and bidirectional DNA sequencing.A heterozygous c390_395del (p.Gln130_Gln131del) was detected. This novel deletion results in the in-frame loss of two glutamine residues in exon 2 of the HNF1B protein. These residues are highly conserved and located in the HNF1 N-terminal domain that contains a dimerization sequence and an acidic region that may be involved in transcription activation. Parental testing has determined that this variant is likely de novo.This de novo variant has not been previously described, hence its pathogenicity is unknown. However the phenotype is consistent with RCAD (MODY5).Written informed consent was obtained from the patient for publication of this Case report. A copy of the written consent is available for review by the Editor-in-Chief of this journal."} +{"text": "Previous studies suggest that extensive heating and food matrix diminish the allergenicity of egg white (EW) proteins, making it possible to be tolerated by some allergic patients.22 children were included: 18 with history of immediate reaction and 4 sensitized to egg (EW sIgE > 50 KU/L). All 22 underwent open OFC with BE performed in 3 days. Day one: OFC with cookies (brand containing egg). Tolerant patients incorporated cookies and food containing egg traces into their diet. A second challenge with home-breaded chicken was performed a week later. Regular consumption was advised for tolerant kids and 15 days later they were challenged with a serving size of home-made cake containing 3 eggs. All children continued regular ingestion and were periodically controlled. Factors including SPT and sIgE levels were used to determine a subsequent challenge with less-heated-egg.All patients tolerated cookies. A 15 month old boy presented mild anaphylaxis (MA) during OFC with breaded chicken (exercise cofactor). Afterwards, he tolerated it and 1 year later passed the OFC with raw egg. A 14 month old girl presented MA during OFC with cake. She continued regular intake of breaded foods and cookies and tolerated omelet 12 months later. All the remaining patients tolerated BE without reactions. In the next 3-12 months 20/22 patients were successfully challenged with hard-boiled egg, 19/22 were challenged with omelet (16 passed/1 MA/ 2 cutaneous symptoms) and 3/22 with raw egg (2 passed/1 cutaneous symptoms). During home dosing only mild symptoms were observed in few patients.BE is well tolerated and safe. Thus, strict dietary avoidance may not be necessary, even from the diagnosis moment. Regular ingestion of HE could also change the natural course of egg allergy."} +{"text": "Melanocortin receptors (MCRs) are seven transmembrane G protein-coupled receptors that are known for their broad physiological relevance. The subtype 4 melanocortin receptor (MC4R) has emerged as a central element in the regulation of energy homeostasis, eating behavior and regulation of sexual functions. MCRs are governed by a complex dynamic homotropic regulation . There i"} +{"text": "Here, we have used transgenesis in zebrafish in order to define regulatory elements that drive lhx5 expression in the forebrain. Through comparative genomic analysis we identified 10 non-coding sequences conserved in five teleost species. We next examined the enhancer activities of these conserved non-coding sequences with Tol2 transposon mediated transgenesis. We found a proximately located enhancer gave rise to robust reporter EGFP expression in the forebrain regions. In addition, we identified an enhancer located at approximately 50 kb upstream of lhx5 coding region that is responsible for reporter gene expression in the hypothalamus. We also identify an enhancer located approximately 40 kb upstream of the lhx5 coding region that is required for expression in the prethalamus . Together our results suggest discrete enhancer elements control lhx5 expression in different regions of the forebrain.The LIM homeobox family protein Lhx5 plays important roles in forebrain development in the vertebrates. The Lhx5 prx domain . LHX5 geophrenia , 4. A fuult mice . Lhx5 re regions \u201313. Prev Xenopus , promote Xenopus , and reg Xenopus .lhx5 gene has complex temporal and spatial expression patterns during embryonic development. LHX5 transcripts are detected in human fetal brain and in various regions of adult central nervous system including the spinal cord, thalamus, and cerebellum [Lhx5 expression is detected in the most anterior portion of the neural tube at the headfold stage. After neural tube closure, Lhx5 is expressed within diencephalic primordium. By mid-gestation, Lhx5 is expressed in the diencephalon and ventral telencephalon. Lhx5 is also expressed in the midbrain, hindbrain, and spinal cord after E10.5 of gestation [Xenopus lhx5 is expressed in entire ectoderm in early gastrula embryo. During neurulation, expression of the lhx5 gene is rapidly restricted to an anterior region in the developing neural plate/keel. In 2-day old Xenopus embryo, this region is more sharply defined, forming a strongly lhx5-expressing domain in the diencephalon anterior to the midbrain-forebrain boundary [lhx5 expression patterns resemble those of Xenopus and by the segmentation stage, zebrafish lhx5 transcripts are detected in the telencephalon, diencephalon, and discrete regions in the hypothalamus and hindbrain [The rebellum . In mousestation . Compareboundary , 20. Zebindbrain .lhx5 genes, the mechanisms underlying lhx5 transcriptional regulation are not well understood. We showed previously a modified bacteria artificial chromosome (BAC) transgenic zebrafish lhx5 line recapitulated endogenous lhx5 expression patterns [lhx5 genomic sequences from various vertebrate species. We find multiple conserved non-coding sequences (CNSs) among different species. We further examine the transcriptional regulatory activity of these CNSs by transient and stable transgenic methods in the zebrafish model.Despite these broadly conserved expression patterns of patterns , 22. Thulhx5 loci to look for CNSs with putative roles in lhx5 gene transcriptional regulation. Twelve vertebrate species representing major groups of the jawed vertebrate lineage were included in our analyses, including genomic sequences from human, mouse, chicken, frog, coelacanth, elephant shark, spotted gar, and five teleost fishes are evolutionary conserved intergenic or intronic sequence elements derived from a common ancestor. We carried out comparative sequence analysis of the t fishes . The sha species . Thus thlhx5 gene upstream genomic sequences [lhx5 genomic region contained the two additional tetrapod specific CNSs and four of the eight teleost specific CNSs, in addition to the two common CNSs conserved in all 12 species and CNS9 both gave rise to robust reporter EGFP expression in the forebrain regions including the telencephalon and the retina and generated stable transgenic animals. The mutated transgenes were mapped to chromosome 13 and chromosome 24 in two independent stable transgenic lines, respectively : chromosome 21:14116198..14290982; Takifugu rubripes (fugu): chromosome 6:5719292..5770520; Oryzias latipes (medaka): chromosome 12:2078644..2245397; Oreochromis niloticus (Nile tilapia): NC_022205:11084704..11177152; Lepisosteus oculatus (spotted gar): NC_023198:14084503..14284612; Astyanax mexicanus : NW_006749407:14473..208111; Latimeria chalumnae (coelacanth): NW_005819674:109551..725575; Gallus gallus (chicken) chromosome 15:12452471..12575304; Homo sapiens (human) chromosome 12:113462889..113966371; Mus musculus (house mouse) chromosome 5:120116513..120441457; Xenopus tropicalis (western clawed frog) NW_004668232:61749600..61904154; Callorhinchus milii (elephant shark) NW_006890147:2032526..2289716. Alignments of long genomic sequences were performed with the mVISTA LAGAN program [lhx5 at the nucleotide level the ClustalX program [Sequences for 12 gnathostome program , 49. ForZebrafish were maintained in a recirculating water system according to standard protocol . ZebrafiThe backbone plasmid pminiTol2-super-Gal4FF contains minimal Tol2 transposable elements flankingTo divide the prethalamic specific CNS4 element into four overlapping fragments, PCR primers listed in zic binding site, we replace the original binding site sequence GACCTCCC with GAAATAAA via chemical synthesis of the ~200 bp F3 fragment.To mutate the lhx5 CNS were prepared over standard columns then further purified by dialysis. Tol2 transposase mRNA was in vitro transcribed from linearized pCS2-TP plasmid (kindly provided by Prof. Koichi Kawakami) with mMESSAGE mMACHINE SP6 Transcription Kit and then column-purified. For injection, equal volumes of 20 ng/\u03bcl of plasmid DNA and 200 ng/\u03bcl of transposase mRNA were mixed and 1~2 nl of the mixture was injected into one-cell stage embryos.Generation of transgenic zebrafish was performed essentially as described . BrieflyTo identify transgenic founder fish, 50\u2013100 of embryos injected with a given plasmid were raised to maturity and outcrossed with Tg(UAS:EGFP) reporter line animal . OutcrosZebrafish embryos were treated with 0.003% 1-phenyl-2-thiourea (PTU) to inhibit pigmentation. Embryos were anesthetized with tricaine, mounted in 3% methyl cellulose, and imaged under a fluorescence microscope (Leica M205FA).Whole-mount in situ hybridization was performed as described . DigoxigTo make probe clones, RT-PCRs were performed with primers listed in To block Fgf signaling, embryos were treated with 20 \u03bcM SU5402 in embryos medium from 10 hpf to 24 hpf at 28.5\u00b0C. Control embryos were treated in embryo medium with DMSO (SU5402 vehicle). Embryos were placed in 12-well plates (30 embryos per well) with 0.5 ml of medium. After drug treatment, embryos were fixed and processed for in situ hybridization.To determine insertion sites of the transgenic enhancer constructs, high efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR) was performed as described , with prS1 FigVista plot of the aligned sequences. Zebrafish sequence is used as the base. The identified CNSs are labeled on the Vista plot.(TIFF)Click here for additional data file.S2 FigA) Figure panels illustrate established transgenic lines carrying corresponding CNS elements. Line id is indicated on the upper left corner on the figure panel. Lateral view of the forebrain regions of embryos, anterior to the left. (B) Text description of the established transgenic lines. The line id used in experiments described in the Result section is indicated.((TIFF)Click here for additional data file.S3 FigThe mutated transgene is inserted at chr13:10057513 and chr24: 1892994 in two independent stable transgenic lines, respectively. The coordinates are based on the Ensembl zebreafish GRCz10 genomic sequence build.(TIFF)Click here for additional data file.S1 TableThe sequence coordinates on the Ensembl zebreafish GRCz10 genomic sequence build and the BAC DKEY-106C24 sequence are listed. The sequence identities of CNS elements between the GRCz10 build and DKEY-106C24 are also listed.(XLSX)Click here for additional data file.S2 TableThe sequence coordinates and sizes of the cloned fragments are listed on sheet named CNS. The primer used in sub-dividing CNS4 are listed on sheet named CNS4.(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file."} +{"text": "Muscle metaboreceptors are afferent signals from working muscles that enhance sweating and attenuate cutaneous vasodilation during a passive heat stress in humans . HoweverStudy \u0399: Fourteen young adults were passively heated for 15.5 min using a upper body water perfused suit (34 \u00b0C) and immersing the participant's legs in hot water (43 \u00b0C). During the heating period, the participants performed 1.5 minutes of isometric hand-grip exercise at 40 % of maximum voluntary contraction with or without (Control) post exercise occlusion of the limb with a pressure cuff to stimulate muscle metaboreceptors for 9 minutes. Study \u0399\u0399: Twelve (6 for each sex) young subjects performed the similar forearm muscle metaboreceptors stimulation while cycling for 13.5 minutes at the exercise intensity of 40 % maximum oxygen consumption while wearing the water perfused suit.tudy \u0399: Forearm muscle metaboreceptors stimulation significantly lowered \u0394mean body temperature thresholds for sweating and cutaneous vasodilation on the oppositional arm of hand-grip exercise during passive heating compared with Control (P <0.05) without differences in thermosensitivity of the responses. Study \u0399\u0399: Forearm muscle metaboreceptors stimulation did not significantly (P > 0.05) affect the onset threshold and thermosensitivity for sweating and cutaneous vasodilation during exercise.SOur results suggest that forearm muscle metaboreceptors activation can modulate heat loss responses during a passive heat stress only. The acceleration of sweating and cutaneous vasodilation through a reduction in the core temperature thresholds of the responses would suggest a central modulation of temperature regulation . It is tForearm muscle metaboreceptors activation lowers core temperature thresholds for heat loss responses during passive heating but not during exercise."} +{"text": "Monogenic autoinflammatory diseases present with overlapping clinical features such as recurrent fever, biological inflammation, abdominal pain, arthritis, and sometimes disabling complications. Over 30 genes have been confirmed or hypothesized as causing diseases. Sanger sequencing is the gold-standard approach for genetic diagnosis, but cannot be exhaustive.We aimed at offering a quick and efficient service for these genetically heterogeneous disorders through NGS sequencing of all published and candidate (N=32) autoinflammatory genes.34 patients were selected on clinical grounds by members of the French reference center for autoinflammatory diseases (CeReMAI). 9 of them were considered positive control samples as they had previously known variants (by Sanger sequencing) and were run in parallel for methodological validation.A custom panel of 108kb was designed to capture the genomic regions of interest using the Illumina Nextera Rapid Capture enrichment DNA preparation kit. MicroV2 chip were then loaded with 12 samples on a MiSeq equipment for multiparallel sequencing. Annotation and filtering were performed with Seqnext software (JSI), and annotation was performed with Alamut Visual (Interactive Biosoftware).CARD14, NOD2, PSTPIP1 and SCL29A3. About 20 new potentially pathogenic sequence variants were found in published genes.We obtained a 545X average coverage and only 2 exons were not captured. Minimal coverage was 40X. Results were concordant in 100% of the cases with Sanger analysis. We uncovered pathological mutations or variants of unknown significance in 17 patients. Of note recurrent genes were This innovative sequencing approach demonstrated high performance in evaluating mutations in known autoinflammatory genes. The current positive rate using sequential Sanger sequencing of 1-4 genes in the French network is quite stable around 10-15%. The NGS approach allowed diagnosis of approximately another 20% of patients."} +{"text": "Development of EMT and consequent activation of stemness programming is responsible for invasion, tumor self-renewal and drug resistance leading to breast cancer progression, distant metastases and poor prognosis. In this study, we employed the luminal ER+ MCF-7 and the IBC SUM149PT breast cancer cell lines to establish the extent to which high grade of CIN and chemoresistance were mechanistically linked to the enrichment of CD44+/CD24low/\u2212 CSCs. Here, we demonstrate that SUM149PT cells displayed higher CIN than MCF-7 cells characterized by higher percentage of structural and numerical chromosomal aberrations. Moreover, centrosome amplification, cyclin E overexpression and phosphorylation of retinoblastoma (Rb) were restricted to the stem-like CD44+/CD24\u2212/Low subpopulation isolated from SUM149PT cells. Significantly, CD44+/CD24\u2212/Low CSCs displayed resistance to conventional chemotherapy but higher sensitivity to SU9516, a specific cyclin-dependent kinase 2 (Cdk2) inhibitor, demonstrating that aberrant activation of cyclin E/Cdk2 oncogenic signaling is essential for the maintenance and expansion of CD44+/CD24\u2212/Low CSC subpopulation in IBC. In conclusion, our findings propose a novel therapeutic approach to restore chemosensitivity and delay recurrence of IBC tumors based on the combination of conventional chemotherapy with small molecule inhibitors of the Cdk2 cell cycle kinase.Inflammatory breast cancer (IBC) is an angioinvasive and most aggressive type of advanced breast cancer characterized by rapid proliferation, chemoresistance, early metastatic development and poor prognosis. IBC tumors display a triple-negative breast cancer (TNBC) phenotype characterized by centrosome amplification, high grade of chromosomal instability (CIN) and low levels of expression of estrogen receptor \u03b1 (ER\u03b1), progesterone receptor (PR) and HER-2 tyrosine kinase receptor. Since the TNBC cells lack these receptors necessary to promote tumor growth, common treatments such as endocrine therapy and molecular targeting of HER-2 receptor are ineffective for this subtype of breast cancer. To date, not a single targeted therapy has been approved for non-inflammatory and inflammatory TNBC tumors and combination of conventional cytotoxic chemotherapeutic agents remains the standard therapy. IBC tumors generally display activation of epithelial to mesenchymal transition (EMT) that is functionally linked to a CD44 Inflammatory breast cancer (IBC) is an angioinvasive form of breast cancer in which cancer cells block the lymphatic vessels in the skin covering the breast and it is associated with a high incidence of early nodal and systemic metastases . This ty+/CD24low/\u2212 phenotype with increased capacity for self-renewal, invasion, drug resistance and tumor progression , centrosome amplification and genomic instability. Significantly, CD44+/CD24low/\u2212 CSCs displayed higher sensitivity to a specific Cdk2 inhibitor than the bulk SUM149PT cells indicating that aberrant activation of cyclin E/Cdk2 oncogenic signaling is essential for the maintenance and expansion of CD44+/CD24low/\u2212 CSCs subpopulation in IBC. In conclusion our findings highlight a novel therapeutic approach based on the combination of conventional chemotherapy with small molecule inhibitors of the Cdk2 cell cycle kinase to treat chemoresistant IBC tumors.In this study we employed the luminal ER2 atmosphere.The human breast cancer cell line MCF-7 was obtained from ATCC , normal human mammary epithelial cells HMEC and IBC SUM149PT cancer cells were kindly provided by Dr Lingle and Dr Couch\u2019s laboratories, respectively . All cell lines were maintained in EMEM medium containing 5 mM glutamine, 1% penicillin/streptomycin, 20 microgram insulin/ml and 10% FBS at 37\u00b0C in 5% CO\u00ae probe were performed as recommended by the manufacturer. Image acquisition and spectral analysis of metaphase cells were achieved by using the SD200 SpectraCube\u2122 Spectral Imaging system mounted on a Zeiss Axioplan2 microscope . Images were analyzed using HiSKY analysis software .Cell harvest and metaphase slide preparation for routine cytogenetic and spectral karyotyping (SKY) analysis were performed as previously described \u201325. Hybr+/CD24\u2212 breast cancer initiating cells by FACS sorting analysis as previously described were employed to identify and isolate CD44escribed .Immunoblot and immunofluorescence studies were performed as previously described . Antibod4 cancer cells were plated in 6-well costar plates and cultured in complete EMEM medium for 48 h. Following 48-h incubation, cancer cells were treated with 1 \u03bcM methotrexate, 0.5 \u03bcM paclitaxel alone or in combination with 1 \u03bcM SU9516 (Sigma) for additional 72 h. Cytotoxicity of conventional chemotherapy alone and in combination with SU9516 was tested by immunofluorescence employing a cleaved-PARP antibody as a marker of apoptosis. Results are derived from three independent experiments.For chemoresistance studies, 5\u00d710+ breast tumors, we employed luminal ER\u03b1+ MCF-7 and IBC SUM149PT cancer cell lines to investigate their level of chromosomal abnormalities; normal mammary epithelial HMEC cells were used as control. The SUM149PT cancer cell line was isolated from an IBC tumor and carries the 2288delT mutation that is linked to loss of BRCA1 function and represents an excellent preclinical model to study the molecular mechanisms responsible for the development of chemoresistance and tumor progression in IBC tumors or SU9516 and cytotoxicity was quantified by analyzing the percentage of cells harboring cleaved PARP as a marker of apoptosis. Significantly, CD44+/CD24\u2212/Low CSCs displayed a higher resistance to methotrexate while they where highly sensitive to the Cdk2 inhibitor SU9516 compared to bulk SUM149PT cancer cells and a specific Cdk2 inhibitor (SU9516) to abrogate the cyclin E/Cdk2 oncogenic signaling. CSCs showed higher resistance to methotrexate than bulk SUM149PT cells, nonetheless CSCs were more sensitive to the cytotoxic effects of SU9516 indicating that molecular inhibition of Cdk2 kinase activity selectively targeted CSCs in IBC. Finally, we showed that combination of paclitaxel with SU9516 in vitro induces a stronger cytotoxic effect characterized by activation of apoptosis in SUM149PT cells. Importantly, the preclinical relevance of our findings is justified by recent studies demonstrating that administration of cyclin E siRNA in vivo inhibited breast tumor growth in nude mice. Moreover, the authors demonstrate that cyclin E siRNA synergistically enhanced the cell killing effects of chemotherapeutic agents in vitro and this combination greatly suppressed the tumor growth in nude mice. In conclusion our study demonstrate for the first time that aberrant activation of cyclin E/Cdk2 oncogenic signaling is restricted in CSCs derived from IBC cells and combination of conventional chemotherapy with small molecule inhibitors of Cdk2 kinase activity may represent a novel targeted therapeutic approach to treat aggressive IBC tumors with consequent benefits on the disease-free and overall survival of patients.The findings presented in this study demonstrate that IBC SUM149PT cancer cells exhibit higher CIN based on structural and numerical chromosome abnormalities than luminal ER\u03b1 and CIN . Signifi and CIN . Concurr and CIN . Because"} +{"text": "Patients receiving anthracyclines (A) and herceptin (H) for treatment of breast cancer require serial monitoring of LV function. Although CMR is considered the gold standard for assessment of volumes and function, most tertiary institutions rely on echocardiography or gated heart pool scans. 2D transthoracic echocardiographic (TTE) image quality relies on appropriate acoustic windows, and accuracy and reproducibility may be particularly limited in this population due to mastectomy and scar. We sought to assess the utility of TTE (LVEF and GLS) compared to CMR in the diagnosis of Stage B heart failure in breaIn total, 44 patients (30 receiving A and 14 receiving T) underwent cardiac magnetic resonance (CMR) for LV function and advanced 2D echocardiography for LV function and global longitudinal strain (GLS) at 3 time-points . We defined stage B heart failure as LVEF < 10% from baseline, and/or LVEF below 55% and/or GLS < -19.7[CMR cine assessment of LV function was performed in all 44 patients (100%). Thirty percent (13/44) could not have accurate echocardiographic EF assessment performed at 1 or more time points. Early LV dysfunction was detected by CMR following commencement of chemotherapy (table). There was poor correlation between CMR LVEF and echocardiography EF (graph) with wide limits of agreement (-23.526 to 13.015%). At 12 months, 8 patients (18%) demonstrated stage B heart failure, however none were below the lower limits of normal (>55%). None of these showed a significant change in TTE EF, however 2 of these patients had GLS < 19.7 at 12 months.A significant number (approx. 1/3) of breast cancer patients are unable to have adequate LV function assessment by echocardiography. Subtle LV dysfunction may be a precursor to overt cardiomyopathy and is currently not adequately diagnosed by echocardiography even with inclusion of 2D global longitudinal strain. Serial monitoring of LV function using a non-radiation imaging technique is best performed by CMR.None."} +{"text": "The cytoplasmic S100 proteins derived from cells of myeloid origin. Calprotectin (MRP8/14 protein complex) might be a biomarker either for autoinflammation and autoimmunopathy. Since autoinflammatory diseases might be a diagnostic challenge calprotectin may be helpful in the diagnosis of autoinflammatory diseases. Chronic nonbacterial osteomyelitis (CNO) is an autoinflammatory, noninfectious disease. CNO describes a wide spectrum from a monofocal bone lesion to the chronic recurring multifocal osteomyelitis (CRMO). Laboratory and histopathological findings are nonspecific. In some patients systemic inflammatory signs such as elevated acute phase proteins cannot be found.To test the ability of Calprotectin (MRP8/14 protein complex) serum concentrations to monitor disease activity in patients with CNO.Serum concentrations of Calprotectin (MRP8/14 protein complex) in a patient with CNO were determined by a sandwich ELISA.Calprotectin (MRP8/14) level were raised heralding active disease when acute phase proteins . The calprotectin level was 7872,7 ng/ml .Calprotectin (MRP8/14) serum concentrations correlate closely with disease activity and may herald a flare before clinical manifestation. Therefore MRP8/14 serum concentrations are a biomarker indicating disease activity in CNO patients.None declared."} +{"text": "Myco-like viruses have been isolated from fungi, feces of various animals, and plant leaves. We report here the isolation of 3 complete genome sequences of gemycircularvirus-related viruses from healthy bovine serum and human brain and serum samples from patients with multiple sclerosis (MS). Their putative capsid proteins share similarity to Torque teno virus (TTV) open reading frame 1 (ORF1) proteins. A large number of circular single-stranded DNA (ssDNA) viruses were recently identified by metagenomic analyses . The groWe analyzed blood samples taken from healthy cattle and serum samples from patients suffering from multiple sclerosis. The bovine sera were subjected to density gradient centrifugation, followed by rolling circle amplification (RCA) on DNA from single fractions. The fragments obtained after restriction digestion (BamH1 and EcoR1) were cloned into vector pUC19 and sequenced by primer walking . Both enThe genome organization of all 3 isolates revealed a putative spliced replication protein encoded on the negative strand and the coat protein (CP) on the positive strand. The CP was arginine rich, and a similarity to the Torque teno virus (TTV) open reading frame 1 (ORF1) protein was indicated in a DomainSweep analysis . The putThe isolation of these 3 gemycircularvirus-related genomes directly from animal and diseased human tissue poses the question of whether they have some etiological relevance for diseases, such as multiple sclerosis.LK931483, LK931484, and LK931485, respectively.The complete sequences of HCBI8.215, HCBI9.212, and MSSI2.225 have been deposited in the EMBL Databank under accession numbers"} +{"text": "Patients with aortic dilation often present an eccentric transvalvular flow jet. The angle of the flow jet from the aorta centerline, or the flow jet angle (FJA), has been reported as a risk factor in bicuspid aortic valve patients . In receThis study included 40 patients with aortic dilation and aortic tricuspid valves participants . Mid-ascending aorta (MAA) diameter and transvalvular peak velocity (Vpeak) were used to assess aortic dilation, aortic stenosis severity , and classify patients into four groups: Group 1 (MAA<35 mm); Group 2 (35 mm35 mm); Group 4 (MAA>35 mm and AS). 4D flow MRI was performed at 1.5T and 3T systems with full thoracic aorta volume coverage in a sagittal oblique slab . 4D flow data were used to compute a PC-MRA image and aorta volume segmentation was performed using Mimics . The isolated aorta segmentation was used to automatically compute the vessel centerline, to mask 4D flow data, and compute 3D JSLD structure using Matlab . FJA workflow is summarized on Figure Patient characteristics and measurements are presented in Table The assessment of FJA can be automated using the volumetric information of 3D JSLD structure which relates the 3D JSLD structure to the aorta centerline, as obtained from 4D flow data. FJA was significantly higher in patients with severe aortic dilation and concomitant AS. Future longitudinal studies are needed to evaluate the impact of FJA in aortic dilation severity and altered flow patterns.Grant support by NIH R01HL115828, NUCATS Dixon Award, AHA 13SDG14360004. CONACyT postdoctoral fellow grant (223355)."} +{"text": "Ticks (Family Ixodidae) transmit a variety of disease causing agents to humans and animals. The tick-borne flaviviruses are a complex of viruses, many of which cause encephalitis and hemorrhagic fever, and represent global threats to human health and biosecurity. Pathogenesis has been well studied in human and animal disease models. Equivalent analyses of tick-flavivirus interactions are limited and represent an area of study that could reveal novel approaches for TBF control.Ixodes scapularis (Lyme disease tick) embryonic ISE6 cells following infection with Langat virus (LGTV) and identify proteins associated with viral infection and replication. Maximal LGTV infection of cells and determination of peak release of infectious virus, was observed at 36 hours post infection (hpi). Proteins were extracted from ISE6 cells treated with LGTV and non-infectious (UV inactivated) LGTV at 36 hpi and analyzed by mass spectrometry. The Omics Discovery Pipeline (ODP) identified thousands of MS peaks. Protein homology searches against the I. scapularis IscaW1 genome assembly identified a total of 486 proteins that were subsequently assigned to putative functional pathways using searches against the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. 266 proteins were differentially expressed following LGTV infection relative to non-infected (mock) cells. Of these, 68 proteins exhibited increased expression and 198 proteins had decreased expression. The majority of the former were classified in the KEGG pathways: \u201ctranslation\u201d, \u201camino acid metabolism\u201d, and \u201cprotein folding/sorting/degradation\u201d. Finally, Trichostatin A and Oligomycin A increased and decreased LGTV replication in vitro in ISE6 cells, respectively.High resolution LC-MS/MS was used to analyze the proteome of In vitro assays using small molecules implicate malate dehydrogenase (MDH2), the citrate cycle, cellular acetylation, and electron transport chain processes in viral replication. Proteins were identified that may be required for TBF infection of ISE6 cells. These proteins are candidates for functional studies and targets for the development of transmission-blocking vaccines and drugs.Proteomic analyses revealed ISE6 proteins that were differentially expressed at the peak of LGTV replication. Proteins with increased expression following infection were associated with cellular metabolic pathways and glutaminolysis. I. scapularis ISE6 cell line infected with the TBF Langat virus (LGTV) to identify proteins and biochemical pathways affected by viral infection. An LC-MS/MS approach was used to identify proteins that were subsequently assigned to putative cellular pathways based on orthology to proteins in the KEGG database. Biochemical pathways common among arthropods in response to infection with flavivirus and possibly unique to tick-flavivirus interactions, were identified. In vitro cellular assays using small molecules suggest the involvement of the ISE6 proteins, malate dehydrogenase (MDH2), and mitochondria in viral replication. These analyses provide a basis for further studies to identify tick proteins associated with viral replication that could be targeted to disrupt TBF transmission.High-throughput proteomics offers an approach to evaluate changes in cell protein levels following arboviral infection. Research to understand the molecular basis of human-flavivirus interactions has advanced significantly over the past decade, but comparatively little is known regarding interactions between ticks and tick-borne flaviviruses (TBFs). Here, we employed a proteomics approach using an Of these, Kyasanur Forest Disease virus (KFDV) is responsible for an estimated 400\u2013500 human cases per year in India ) help to identify cellular pathways in I. scapularis (genome.jp/kegg/ko). To be identified in a KEGG pathway, KO is required. ISE6 proteins with KO and not identified in I. scapularis (KEGG) pathways are also included. (B) Percent cellular function distribution of proteins found in the 66 identified I. scapularis (KEGG) pathways with 16 modules.(A) (TIF)Click here for additional data file.S4 FigExpression of ISE6 proteins with (A) or without (B) orthology and no identified pathways. Refer to (TIF)Click here for additional data file.S5 Fig\u03b1 S5 Fig, S7 Fig, and S11 Fig of Khadka et al. [\u03bc\u03b2Tables 1 and 2 of Pastorino et al.[\u0394Corresponding percentages correspond to the number of tick ISE6 orthologs identified with orthologs identified in: a et al. ; \u03bcS2 Tab et al. \u03bc; \u03b2Tablesno et al.; \u0394S1 Tabo et al.\u0394.(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 TableI. scapularis ISE6 proteins is based off of \u22651 peptide identification and \u22651 statistical analysis (p < 0.05) identification . From The total number of (XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(DOCX)Click here for additional data file.S5 TableI. scapularis cellular pathways, and ISE6 ortholog proteins with mapped cellular pathways in other eukaryotes. This table is organized into these four groups including protein cellular function, protein class, and protein pathway. Fold changes of LGTV/mock and UV-LGTV/mock (\u201cnd\u201d denotes not detected) are listed along with search results as to whether the protein has been identified in other flavivirus-host proteomic studies. Proteins listed from Figs As mentioned in (XLSX)Click here for additional data file."} +{"text": "Sonic Hedgehog (Shh) signaling is one key pathway that regulates multiple aspects of craniofacial development, but whether it co-ordinates cranial neural crest cell and placodal cell interactions during cranial ganglia formation remains unclear. In this study we examined a new Patched1 (Ptch1) loss-of-function mouse mutant and characterized the role of Ptch1 in regulating Shh signaling during cranial ganglia development. Wig/ WigPtch1 mutants exhibit elevated Shh signaling in concert with disorganization of the trigeminal and facial nerves. Importantly, we discovered that enhanced Shh signaling suppressed canonical Wnt signaling in the cranial nerve region. This critically affected the survival and migration of cranial neural crest cells and the development of placodal cells as well as the integration between neural crest and placodes. Collectively, our findings highlight a novel and critical role for Shh signaling in cranial nerve development via the cross regulation of canonical Wnt signaling.Cranial nerves govern sensory and motor information exchange between the brain and tissues of the head and neck. The cranial nerves are derived from two specialized populations of cells, cranial neural crest cells and ectodermal placode cells. Defects in either cell type can result in cranial nerve developmental defects. Although several signaling pathways are known to regulate cranial nerve formation our understanding of how intercellular signaling between neural crest cells and placode cells is coordinated during cranial ganglia morphogenesis is poorly understood. The cranial nerves are part of the peripheral nervous system that governs various critical functions such as sensing and controlling movement within the craniofacial region. Previous studies in avian embryos have shown that some of the cranial nerves including the trigeminal (V) and facial nerves (VII) originate from both cranial neural crest cells and ectodermal placode cells ,2. CraniWiggable (Wig) that carries a mutation in the Patched1 (Ptch1) gene. Ptch1 encodes a receptor for the Hedgehog family of morphogens which includes Sonic Hedgehog (Shh). Unlike Ptch1 null mutant mice which are lethal at E9.5 [Ptch1Wig/ Wig mutants survive until E12.0, allowing an analysis of the effects of aberrant Shh signaling on cranial ganglia morphogenesis. In this study, we took advantage of multiple mouse mutants to clarify the role of cross-talk between the Shh and WNT signaling pathways during the formation of the trigeminal and facial nerves. We discovered that elevated Shh signaling restricts canonical Wnt signaling during cranial ganglia development. This affects the survival of migrating neural crest cells, the pattern of placode development and the integration between neural crest cells and placode cells. Our findings describe the importance of cross-talk between Shh and Wnt signaling in regulating tissue interactions during cranial nerve development.In a previous study, we performed an N-ethyl-N-nitrosourea (ENU) mutagenesis screen in mice and identified multiple recessive alleles important for craniofacial development . Here weThis study was carried out in accordance with recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol (2013\u20130115) was approved by the Institutional Animal Care and Use Committee of The Stowers Institute for Medical Research. Adult mice were euthanized via CO2 and cervical dislocation according to the recommendations of the American Veterinary Medical Association and all efforts were made to minimize any potential suffering.Ptch1Wig, Ptch1LacZ, Wnt1Cre, R26RYFP, TOPgal and HhatCreface mice were maintained as described previously [Ptch1Wig/Wig as Wig homozygous mutants and HhatCreface/Creface;Ptch1Wig/Wig as double-homozygous mice. Either wild-type or heterozygous littermates were used as control mice described in this study. Unless otherwise indicated, we used a minimum of 4 or 5 embryos from multiple distinct litters for each parameter analyzed in this study.eviously \u201314. The Wiggable (or Wig) mutation was generated in a previously described N-ethyl-N-nitrosourea (ENU) screen for recessive mutations leading to craniofacial and neural tube defects [Wiggable to reflect the superficial resemblance to Baroque period English wigs. DNA from mutant and control littermate embryos was collected along with tail biopsies from founder males and subjected to microsatellite and single nucleotide polymorphism (SNP) mapping, which narrowed down the affected region to mouse chromosome 13 (qB2 to qB3). This region is gene dense but contained only a few genes known to regulate embryonic neural development, including Ptch1. We used a candidate approach to sequence the genomic (intron/exon) regions of the genes residing in the minimal interval. Sequencing primers were designed to cover the entire mouse Ptch1 gene (available upon request), which was one of our prioritized gene candidates in the minimal affected region, and we discovered a T to A substitution in the 3\u2019end of intron 15 which segregated with the mutant phenotype (The defects . Brieflyhenotype .Ptch1 gene. Amplicon length was 92 base pairs (bp) for the wild type product and 99bp for the Wig mutant product. The following primers were used to generate the amplicon: CAA GCA ACT TCC CCA AAT GTG and ATC CTC CCA GTT TCC CAG TCA. Actin endogenous control probe and primers were ordered as a pre-designed assay from Applied Biosystems Endogenous Control # 4352933E). For the mutation site, the following dye-coupled sequences were used: Ptch1 wild type probe: 6FAM\u2014TGG CTT CAA GGA CTT C\u2014MGBNFQ; Ptch1Wig mutant probe: VIC\u2014ACT GGC TTC AAG TTT CTA GGA\u2014MGBNFQ. The linear dynamic range of the qPCR assay was determined by making serial dilutions of the RNA and performing the reverse transcription (RT) reaction and subsequent amplification using the wild type, mutant and Actin endogenous control probes and primers. For the reverse transcription (RT) reaction, we utilized the high capacity cDNA reverse transcription kit from Applied Biosystems , using 100ng of RNA per reaction in a total volume of 20\u03bcL. Each sample was then split into 3 reactions where the wild type, mutant and endogenous control primers and probes were added. The cycling conditions for the cDNA amplification were: denaturation at 95\u00b0C for 2 min, followed by 40 cycles of 95\u00b0C for 15 seconds (sec) and 60\u00b0C for 1 minute (min). Each sample was genotyped by amplification of genomic DNA with forward and reverse primers detailed above using the following cycling conditions: 95\u00b0C for 2 min then 32 cycles of 95\u00b0C/30sec, 60\u00b0C/30 sec, and 72\u00b0C/30 sec.) The resulting amplicon was sequenced from both directions using the same primers and aligned to the expected sequence to identify the mutation polymorphism and quantified using a Bioanalyzer. Primer Express v3.0 was used to make the primers and probes covering the splice junction between exons 15 and 16 of the morphism .Wiggable (Wig) founders and their offspring confirmed the mutation introduced a 7 base pair insertion into the Ptch1 cDNA to introduce the 7 bp insertion into the full-length mouse Ptch1 cDNA which was cloned in the expression vector pcDNA3.1 (gift from Dr. Kazushi Aoto). The insertion was confirmed by sequencing. Wild type Ptch1 or Ptch1Wig was transfected into Hek293T cell lines and after 48 hours culture, protein was extracted using a standard lysis buffer . Protein was quantified using a BSA curve and 40\u201360ng of protein was loaded per lane. Blots were transferred onto PVDF membranes and blocked with 5% BSA solution. To detect PTCH1 protein, blots were probed with an amino-terminal specific PTCH1 antibody .The sequencing of ch1 cDNA . This inWig mutation was indeed a loss-of-function mutation in Ptch1, we conducted a functional complementation assay with Ptch1LacZ/ + mice (Jackson Laboratory strain 003081). The Ptch1LacZ allele contains a lacZ-neo fusion gene that disrupts endogenous Ptch1 gene function [Ptch1LacZ/ LacZ embryos exhibit neural tube abnormalities and are typically lethal at E9.5. Ptch1Wig/+ heterozygotes were interbred with PtchLacZ/+ heterozygotes to generate embryos which were genotyped for the presence of both Wig and LacZ alleles. \u03b2-galactosidase staining (described below) was evaluated to determine the spatiotemporal expression of the Ptch1 locus in the Ptch1Wig/LacZ mutants.To test whether the function . Ptch1LWhole-mount immunostaining of mouse embryos was performed as previously described ,15 usingin situ hybridization and sectioning was performed as previously described [Whole-mount escribed .TOPgal reporter mice [Ptch1Wig/+ and HhatCreface/+;Ptch1Wig/+ mice and embryos were stained for LacZ activity using the \u03b2-galactosidase Staining Solution Kit (Chemicon/Millipore) according to the manufacturer\u2019s instructions.ter mice were breWiggable (Wig) due to a kinked and overproliferative neural tube that superficially resembled an English wig (Patched1 (Ptch1), which encodes a receptor for Hedgehog ligands [Wig ENU mutants segregated with a T to A substitution in the 3\u2019end of intron 15 of Ptch1, which creates a novel splice acceptor site and produces a 7 base pair insertion in the 5\u2019 end of Exon 16 -terminal-specific Ptch1 antibodies to probe the resulting Western blot, we confirmed that the Wig allele of Ptch1 did indeed produce a stable truncated form of PTCH1 that migrated at \u223c90kDa expressed only the mutant form of Ptch1, with undetectable levels of wild type Ptch1 mRNA expression (Ptch1Wig/+ heterozygotes (Specimens 220\u2013222) showed approximately equal levels of both the wild type and Wig mutant Ptch1 mRNA. No Ptch1Wig mutant mRNA was detected in wild type embryos. Interestingly, in the Ptch1Wig/Wig embryos Ptch1 mRNA levels were highly elevated, consistent with deregulated Shh signaling. This is because the Ptch1 locus is under the direct transcriptional regulation of Shh signaling through the binding of GLI transcription factors to its promoter [The t \u223c90kDa . To furtrcrosses . Ptch1Wpression . Ptch1WPtch1Wig allele with the Ptch1LacZ mutation [Ptch1LacZ mutation creates a null allele that in Ptch1LacZ/LacZ embryos leads to severe patterning defects and lethality at E9.5. In contrast the Ptch1Wig mutation is less severe, with Ptch1Wig/Wig embryos surviving until E11.5\u201312.5 and facial nerves (VII) develop as clusters of Neuronal Class III \u03b2-tubulin (TUJ1) labeled neurons loss-of-function in migrating neural crest cells leads to neural crest cell death [We recently discovered that Shh signaling restricts the extent of canonical Wnt signaling in the developing craniofacial prominences . Furthersurvival . We founreporter . We obsell death . CollectPtch1Wig/Wig mutant embryos. We used PAX3 and SOX10 co-immunostaining to define the placode cells of the trigeminal region [Ngn1, Ngn2, and NeuroD expression via in situ hybridization as indicators of neuronal maturation in the placode [Ptch1Wig/Wig embryos loss-of function mutant (HhatCreface/Creface) to lower the levels of Shh signaling in Ptch1Wig/Wig mutants by producing compound mutants. The disruption of Hhat diminishes palmitoylation of SHH which perturbs its secretion and long range activity [TOPgal reporter into HhatCreface/Creface;Ptch1Wig/Wig compound mutant mice to visualize the spatiotemporal activity of canonical Wnt signaling. In E9.5 Ptch1Wig/Wig embryos, Wnt signaling as measured by TOPgal activity was diminished in the trigeminal (red arrowhead) and facial (white arrowhead) nerve regions or \u03b1N-catenin (Ctnna2) are important for cranial nerve integration [N-cadherin (Cdh2) expression in Xenopus results in abnormal aggregation and migration of neural crest cells [Ptch1Wig/Wig embryos may also be due in part to disrupted Wnt signaling-dependent cell adhesion in cranial neural crest or/and placode cells, in addition to the reduced numbers of neural crest cells that is caused by apoptosis.In this study, we observed relatively poor integration between neural crest and neurogenic cells in the trigeminal and facial nerves of ls Figs. and 7. Regration ,41. Excest cells . This sust cells . CollectElevated Shh signaling results in various well characterized craniofacial anomalies including anencephaly, hyperterolism, and cleft lip and palate ,44,45 asIn conclusion, we have demonstrated a role for cross-talk between Shh and Wnt signaling in neural crest cells and placodes cells during cranial nerve development. Elevated Shh signaling did not affect the formation and migration of cranial neural crest cells, but did result in selective death of migratory neural crest cells. Elevated Shh signaling also perturbed the development of the neurogenic placodes, particularly the trigeminal placode. Together these events disrupted neural crest and placode cell integration and led to aberrant cranial nerve development. Shh signaling restricts canonical Wnt signaling, and reduced Wnt signaling is associated with neural crest cell death and abnormal cranial nerve development. Using genetic approaches to reduce the levels of elevated Shh signaling, we were able to restore the canonical Wnt signaling in the neural crest and placodal territories of the developing head and rescue cranial ganglia morphogenesis.S1 FigPtch1Wig mutation creates a novel consensus splice acceptor site at the 3\u2019 end of intron 15 due to A to T substitution. This in turn leads of a 7 base pair insertion (gtttctag) and premature truncation (tga) 17 base pairs downstream of the 5\u2019 end of Exon 16 of the Ptch1 gene. Sequencing of cDNAs derived from the biopsies of Wig carrier mice confirmed the presence of the predicted 7 base pair insertion.The (TIF)Click here for additional data file.S2 FigPtch1Wig/Wig mutants (red), Ptch1Wig/+ heterozygotes or Ptch1+/+ wild type (grey) E10.5 embryos. Embryos derived from Ptch1Wig/+ intercrosses were lysed and subjected to RT-PCR. Primers specific for the Wiggable mutation site in exon 16 were used in qPCR experiments, with values normalized to \u03b2-actin transcript levels. Values were plotted as +/- standard deviation. Ptch1Wig/Wig mutants did not display significant levels of wild type Ptch1 transcripts, and show an upregulation of Ptch1Wig levels. This upregulated Ptch1 locus activity was only present in Ptch1Wig mutants and not in herteozygotes. (B) Raw data for the Ptch1 wild type and Wig signal obtained from various embryo specimens.(A) Bar chart of the qPCR levels in (TIF)Click here for additional data file.S3 FigPtch1Wig mutant embryos (G and I) in the ophthalmic and facial nerve. There was increased cell death in SOX10-positive migratory neural crest cells in Ptch1Wig mutants relative to controls (C and E). (K) No statistically significant difference in neuronal cell death numbers between control and Ptch1Wig/Wig embryos. (L) Ptch1Wig/Wig embryos showed significantly increased number of apoptotic SOX10-positive neural crest cells in the ophthalmic region relative to controls. Scale bars: 100\u03bcm (A and F); 20\u03bcm ; 50\u03bcm . *P < 0.05, Student\u2019s t test. Data are represented as mean \u00b1 SEM.(A and F) Whole mount immunostaining of Neuronal Class III \u03b2-tubulin (TUJ1) (red) and DAPI (blue). (B-E and G-J) Horizontal sections across the indicated planes in (A) and (F) of the ophthalmic and facial nerve immunostained for TUJ1 or SOX10 , along with TUNEL (green) and DAPI (blue). No difference in cell death between control (B and D) and (TIF)Click here for additional data file."} +{"text": "Mrps34 mutation causes a significant decrease of this protein, which we show is required for the stability of the 12S rRNA, the small ribosomal subunit and actively translating ribosomes. The synthesis of all 13 mitochondrially-encoded polypeptides is compromised in the mutant mice, resulting in reduced levels of mitochondrial proteins and complexes, which leads to decreased oxygen consumption and respiratory complex activity. The Mrps34 mutation causes tissue-specific molecular changes that result in heterogeneous pathology involving alterations in fractional shortening of the heart and pronounced liver dysfunction that is exacerbated with age. The defects in mitochondrial protein synthesis in the mutant mice are caused by destabilization of the small ribosomal subunit that affects the stability of the mitochondrial ribosome with age.The evolutionary divergence of mitochondrial ribosomes from their bacterial and cytoplasmic ancestors has resulted in reduced RNA content and the acquisition of mitochondria-specific proteins. The mitochondrial ribosomal protein of the small subunit 34 (MRPS34) is a mitochondria-specific ribosomal protein found only in chordates, whose function we investigated in mice carrying a homozygous mutation in the nuclear gene encoding this protein. The Mitochondria make most of the energy required by eukaryotic cells and therefore they are essential for their normal function and survival. Mitochondrial function is regulated by both the mitochondrial and nuclear genome. Mutations in nuclear genes encoding mitochondrial proteins lead to mitochondrial dysfunction and consequently diminished energy production, a major symptom of metabolic and mitochondrial diseases. The molecular mechanisms that regulate mitochondrial gene expression and how dysfunction of these processes causes the pathologies observed in these diseases are not well understood. Messenger RNAs encoded by mitochondrial genomes are translated on mitochondrial ribosomes that have unique structure and protein composition. Mitochondrial ribosomes are a patchwork of core proteins that share homology with prokaryotic ribosomal proteins and mitochondria-specific proteins, which can be unique to different organisms. Mitochondria-specific ribosomal proteins have key roles in disease however their functions within mitochondria are not known. Here we show that a point mutation in a mammalian-specific ribosomal protein causes mitochondrial dysfunction, heart abnormalities and progressive liver disease. This mouse provides a valuable model to elucidate the pathogenic mechanisms and progression of metabolic diseases with age, while enabling a more thorough understanding of mitochondrial ribosomes and protein synthesis. Mitochondria are composed of proteins encoded by the nuclear and mitochondrial genomes. Most of the mitochondrial proteins including the ribosomal proteins and translation factors that are responsible for the expression of the mitochondrial genome are synthesized on cytoplasmic ribosomes and imported into mitochondria post-translationally. In chordates, the mitochondrial genome encodes 22 tRNAs, 2 rRNAs and 11 mRNAs that are translated on mitochondrial ribosomes (mitoribosomes) into 13 polypeptides, all members of the oxidative phosphorylation complexes as described before x 100.Fresh sections of the liver and muscle were frozen in Optimal Cutting Temperature (OCT) medium, sectioned and stained with Haematoxylin and Eosin, Gomori\u2019s Trichrome and Haematoxylin, Oil Red O and Haematoxylin, COX or NADH stains. Images were acquired using a Nikon Ti Eclipse inverted microscope using a Nikon 40x objective and Oil Red O staining was quantified as described previously .S1 TextBehavioral and motor testing.(DOC)Click here for additional data file.S1 FigMrps34wt/wt and Mrps34mut/mut mice was determined by immunoblotting.The distribution of MRPS34 and \u03b2-actin in tissue homogenates from (TIF)Click here for additional data file.S2 FigMrps34wt/wt (n = 5) and Mrps34mut/mut (n = 5) by pulse incorporation of 35S-labeled methionine and cysteine after 1 hour. Equal amount of cell lysate protein (50 \u03bcg) was separated on SDS polyacrylamide gels and stained with Coomassie blue. The data shown are representative results of at least five different experiments. (C) Quantification of the rate of translation measured by pulse incorporation of 35S-labelled methionine and cysteine over time of COXIII and ND3 in heart and liver mitochondria from aged mice. Data are means \u00b1 SEM of three separate experiments.Protein synthesis of mitochondrially encoded proteins was measured in heart and liver mitochondria from young (A) and old (B) (TIF)Click here for additional data file.S3 FigMrps34wt/wt and Mrps34mut/mut mice using an OROBOROS oxygen electrode. Data are means \u00b1 SEM of three separate experiments; *, p < 0.05 compared with control treatments by a 2-tailed paired Student\u2019s t test.State 3 and 4 respiration was measured in mitochondria isolated from hearts and livers of young (TIF)Click here for additional data file.S4 FigMrps34wt/wt (n = 5) and Mrps34mut/mut (n = 5) tracking ability using optokinetic drum, measured in number of time spent tracking in seconds. (B) Comparison of Mrps34wt/wt and Mrps34mut/mut tracking ability measured in number of tracks performed. (C) Comparison of time spent in light versus dark in Mrps34wt/wt (n = 5) and Mrps34mut/mut (n = 5) mice measured in seconds. (D) Quantitation of behavioral studies evaluating number of times the box was reached in hanging wire experiments comparing Mrps34wt/wt (n = 5) and Mrps34mut/mut (n = 5) mice. (E) Quantitation of behavioral studies evaluating distance travelled along the wire in hanging wire experiments comparing Mrps34wt/wt and Mrps34mut/mut mice. (F) Rotarod results measured in seconds spent on the rotorrod over 4 days to show improvement and learning ability. (G) Time spent on the rotarod over 4 days to analyze motor function and learning ability. (H) Cresyl violet/toluidine blue staining of optic nerves from Mrps34wt/wt (n = 5) and Mrps34mut/mut (n = 5) mice visualized at 100x magnification. (I) Muscle sections cut at 10 \u03bcm thickness were stained with Haematoxylin and Eosin, COX and NADH from young and aged Mrps34wt/wt (n = 9) and Mrps34mut/mut (n = 9) mice and visualized at 40X magnification.(A) Comparison of (TIF)Click here for additional data file."} +{"text": "The discovery of cell-free fetal DNA (cfDNA) circulating in the maternal blood has provided new opportunities for noninvasive prenatal diagnosis (NIPD). However, the extremely low levels of cfDNA within a high background of the maternal DNA in maternal circulation necessitate highly sensitive molecular techniques for its reliable use in NIPD. In this proof of principle study, we evaluated the earliest possible detection of cfDNA in the maternal plasma by a bead-based emulsion PCR technology known as BEAMing . Blood samples were collected from in vitro fertilization (IVF) patients at 2 to 6 weeks following embryo transfer and plasma DNA was extracted. The genomic regions of both X and Y chromosome-specific sequences (AMELX and AMELY) were concurrently amplified in two sequential PCRs; first by conventional PCR then by BEAMing. The positive beads either for AMELX or AMELY gene sequences were counted by a flow cytometer. Our results showed that the pregnancies yielding boys had significantly higher plasma AMELY gene fractions (0.512 \u00b1 0.221) than the ones yielding girls (0.028 \u00b1 0.003) or non-pregnant women . Here, we clearly demonstrated that the BEAMing technique is capable of reliably detecting cfDNA in the blood circulation of 4-week-pregnant women, which is only two weeks after the embryo transfer. BEAMing technique can also be used to early detect fetal DNA alterations in other pregnancy-associated disorders. The management of these disorders in affected children incurs high financial cost in health care systems and emotional burden on families. The most viable solution in circumventing these problems is through preventive strategies such as pre-implantation genetic diagnosis (PGD) following in vitro fertilization (IVF) and prenatal diagnosis (PND) in spontaneous pregnancies ..20]. TheThe BEAMing technique was originally developed to detect circulating tumor DNA in the plasma of colorectal cancer patients for a noninvasive and more sensitive monitoring of the treatment response to therapy , 22. In We demonstrated that the BEAMing technique is capable of reliably detecting cfDNA in the blood circulation of 4-week pregnant women (two weeks following embryo transfer). This highly sensitive digital assay can also be used in the noninvasive and early detection of the fetal DNA alterations of other pregnancy-associated disorders for possible interventions or terminating pregnancies.S1 FigBeads are separated by a flow cytometer as negative (unamplified), AMELX positive, AMELY positive and double positives (AMELX and AMELY). The beads, positive only for a single signal type are used for calculations; beads with both AMELY and AMELY DNA are excluded from the analyses.(PDF)Click here for additional data file.S2 FigA) 0.0%, (B) 0.01%, (C) 0.1%, (D) 1%, and (E) 10%.The relative AMELY positive fractions of serially diluted samples; ((PDF)Click here for additional data file."} +{"text": "Hospitals\u2019 safety climates have been correlated with incidents of exposure to blood and body fluids.We examined the relationship between laboratory safety climates and hazardous incidents among laboratory personnel at selected health facilities in Kenya.A survey on history of hazardous incidents and safety mechanisms was conducted among laboratory personnel in Kenya. A hazard was defined as a fall, inhalation of harmful gas, ingestion of hazardous agents, subcutaneous chemical exposure, sharp injury or a hazardous spill. The laboratory safety climate was rated on a ten point scale. We used univariate analyses to describe the relationship between experiencing a hazard and the safety climate.One hundred and thirteen laboratory personnel were interviewed; the median duration of service was 4.0 years (range 0.2 - 33 years), 62 (53%) had received Hepatitis B vaccination and 18 (16%) had been previously trained on biosafety. Health facilities were equipped with; sharps\u2019 disposal facilities (90%), PPE (82%), waste disposal mechanisms (60%), containment of hazardous wastes (38%), fire safety equipment (38%), vaccination measures (28%), reporting mechanisms for exposures (22%), safety equipment (19%), protocols for occupational injuries documentation (17%) and safety audits (10%). Eighty-one (77%) laboratory personnel had experienced a hazard. Having been vaccinated against Hepatitis B, trained on biosafety, availed to safety mechanisms and work duration were not associated with history of a hazard.Although the relationship between hazardous incidents and safety mechanisms is not of statistical significance, the absence of appropriate laboratory safety mechanisms it is still of public health concern. Equipping labs with necessary safety infrastructure as well as assessing personal factors related to injuries may decrease the incidences of laboratory hazards.None declared."} +{"text": "GP55 for gB, GP75 for gH, GP115 for gL, GP100 for gM, GP73 for gN and GP74 for gO) in separate reactions was lethal for virus regeneration on fibroblast cells which demonstrated the essential nature of the GPCMV glycoproteins. The gene knockout results were similar to HCMV, except in the case of the gO homolog, which was non-essential in epithelial tropic virus but essential in lab adapted GPCMV. Overall, the findings demonstrate the similarity between HCMV and GPCMV glycoproteins and strengthen the relevance of this model for development of CMV intervention strategies.Development of a cytomegalovirus (CMV) vaccine is a major public health priority due to the risk of congenital infection. A key component of a vaccine is thought to be an effective neutralizing antibody response against the viral glycoproteins necessary for cell entry. Species specificity of human CMV (HCMV) precludes direct studies in an animal model. The guinea pig is the only small animal model for congenital cytomegalovirus infection. Analysis of the guinea pig CMV (GPCMV) genome indicates that it potentially encodes homologs to the HCMV glycoproteins that form various cell entry complexes on the outside of the virus: gCI (gB); gCII (gH/gL/gO); gCIII (gM/gN). The gB homolog (GP55) has been investigated as a candidate subunit vaccine but little is known about the other homolog proteins. GPCMV glycoproteins were investigated by transient expression studies which indicated that homolog glycoproteins to gN and gM, or gH, gL and gO were able to co-localize in cells and generate respective homolog complexes which could be verified by immunoprecipitation assays. ELISA studies demonstrated that the individual complexes were highly immunogenic in guinea pigs. The gO (GP74) homolog protein has 13 conserved N-glycosylation sites found in HCMV gO. In transient expression studies, only the glycosylated protein is detected but in virus infected cells both N-glycosylated and non-glycosylated gO protein were detected. In protein interaction studies, a mutant gO that lacked N-glycosylation sites had no impact on the ability of the protein to interact with gH/gL which indicated a potential alternative function associated with these sites. Knockout GPCMV BAC mutagenesis of the respective glycoprotein genes ( Congenital human cytomegalovirus (HCMV) infection occurs in approximately 1% of live births in the US and can lead to symptomatic disease including mental retardation and hearing loss . In congAnimal model pathogenicity, vaccine and antiviral studies of CMV are carried out with animal-specific CMVs such as guinea pig (GPCMV), mouse (MCMV), rat (RCMV) and rhesus macaques (RhCMV). The genomes of all of these animal CMVs have been sequenced \u201310. The http://www.ensembl.org/Cavia_porcellus/Info/Index) with subsequent follow up with RNA seq analysis which potentially enables the generation of new guinea pig specific reagents. Manipulation of an infectious GPCMV BAC has allowed the preliminary study of some viral genes [A drawback in GPCMV and the guinea pig model has been a lack of development at the molecular level. This has largely been overcome by the recent sequencing of the viral genome and the development of infectious BAC clones of the GPCMV genome , 20, 21.al genes , 22\u201326 bal genes , 28.UL128-131 genes is unstable upon passage of clinical HCMV strains on fibroblast cells and this locus rapidly acquires point mutations or deletions with the subsequent loss of epi/endothelial viral tropism associated with the inability to form a functional complex [In HCMV, a number of proteins have been identified as glycoproteins that are associated with purified virions and extra cellular dense bodies . However complex . The pen complex . A poten complex , 39\u201341. GP75, GP115, GP100, GP73 and GP74 respectively) which are co-linear with their counterparts in the HCMV genome. Presumably, the GPCMV homologs can generate the equivalent HCMV glycoprotein complexes to gCI, gCII and gCIII. Neutralizing antibody responses are generated in GPCMV infection [The essential nature of the GPCMV glycoproteins and their role in the viral life cycle has been largely unexplored with the exception of gB, which has been investigated as a neutralizing vaccine target antigen against congenital CMV \u201345. Howenfection and passnfection . In convGPCMV , first and second generation GPCMV BAC , 21 derihttp://www.cbs.dtu.dk/services/SignalP/ [http://sigpep.services.came.sbg.ac.at/signalblast.html; http://www.csbio.sjtu.edu.cn/bioinf/Signal-3L/ [http://www.cbs.dtu.dk/services/TMHMM/. (3) BLAST alignments performed on MacVector.The predicted protein sequences of the GPCMV glycoproteins were analyzed by various programs. (1) Signal peptide sequence was predicted by various on line programs: SignalP/ ; http://gnal-3L/ . (2) Pro2 overdose followed by cervical dislocation in accordance with IACUC protocol and NIH guidelines.Guinea pig (Hartley) animal studies were carried out under IACUC (Texas A&M University) permit 2013#013. All study procedures were carried out in strict accordance with the recommendations in the \u201cGuide for the Care and Use of Laboratory Animals of the National Institutes of Health.\u201d Animals were observed daily by trained animal care staff, and animals requiring care were referred to the attending veterinarian for immediate care or euthanasia. Terminal euthanasia was carried out by lethal COAll animals were verified by anti-GPCMV ELISA to be seronegative prior to their inclusion in the study. Convalescent antisera to GPCMV was generated by subcutaneous inoculation of GPCMV negative guinea pigs (n = 6) with 1x10^5 pfu salivary gland (SG) stock GPCMV (strain 22122). Seroconverted animals were verified 1 month post infection by anti-GPCMV ELISA. A booster inoculation of virus was given and final bleeds were taken at 8 weeks post initial inoculation. Serum samples were subsequently pooled for studies. Control negative sera was obtained from GPCMV negative animals, verified by ELISA, prior to initial GPCMV challenge.GP55 (gB) 94,164\u201396,869; GP73 (gN) 117,644\u2013118,045; GP74 (gO) 117,992\u2013119,104; GP75 (gH) 119,553\u2013121,724; GP100 (gM) 157,482\u2013158,531; GP115 (gL) 180,216\u2013180,992. All genes are encoded on the complementary strand with the exception of GP73. The predicted GPCMV encoded proteins were verified as homologs of HCMV glycoproteins based on the co-linear location of the GPCMV viral gene with the HCMV genome and the percentage identity of the predicted GPCMV protein with HCMV glycoproteins [GP73 oligos FGP73 & RGP73; for GP74 oligos FGP74 & RGP74; for GP100 oligos GP100F & GP100R; for GP115 oligos FGPgL & RGPgL. The PCR primers also carried an additional 5\u2019 EcoR I restriction enzyme site to introduce flanking EcoR I sites at the ends of the GP73, GP74, GP100 and GP115 coding sequences for ease of cloning of the PCR product. The various ORFs were cloned as EcoR I fragments into pNEB193 except for GP74 which was cloned as an EcoR I fragment into plasmid pNEB193dSalI, which had the Sal I site removed from the poly-cloning region. Individual glycoprotein gene plasmid clones were verified by sequencing. The GP75 ORF was cloned as a Hind III fragment using primers FGP75 and RGP75 tagged with Hind III sites drug resistance marker cassette [Kpn I; EcoR V; Sal I; Nru I; or BamH I. Disruption of each glycoprotein gene on each shuttle vector was carried out as follows. Plasmid pNEBgNT was digested with Nru I and an EcoR V Km cassette was inserted to disrupt the GP73 (gN) ORF at codon 71 (pNEBgNTKm). Plasmid pNEBGP74 was digested with Sal I and a Sal I Km cassette inserted to disrupt the GP74 ORF at codon 110. Plasmid pNEBGP100 was transformed into a dam- strain of E.coli K12 bacteria and cut with Bcl I and a BamH I Km cassette inserted to disrupt the GP100 ORF at codon 110 (pNEBGP100km). Plasmid pNEBgL was cut with EcoR V to insert an EcoR V Km cassette to disrupt GP115 ORF at codon 45 (pNEBgLKm). The GP75 gene in pNEBgH was digested with Nru I and EcoR V to delete 791 bp of the coding sequence. The linearized DNA was band isolated as previously described and a EcoR V Km cassette inserted to disrupt the GP75 (gH) ORF at codon 200 (pUCGP75Km). In the case of GP55, a shuttle vector was designed to encode the homolog gB AD-1 and trans-membrane domains of GP55 [GP55 coding sequence was PCR amplified using primers FgBEc and RgBPst (EcoR I and Pst I restriction sites at the 5\u2019 and 3\u2019 ends of GP55 respectively. The GP55 PCR product was cloned into pUC19 cut with EcoR I and Pst I to generate pUCgB. An EcoR V Km cassette was next introduced into pUCgB at a unique EcoR V site that disrupts the GP55 coding sequence at codon 528. The modified shuttle vector was designated pUCgBKm. The various glycoprotein knockout shuttle vectors (carrying kanamycin insertion markers) were verified by sequencing and subsequently used to create individual glycoprotein gene mutants on the GPCMV BAC in bacteria. Modified plasmids encoding Km cassettes are described in The predicted GPCMV glycoprotein coding sequences were based on the complete viral genome sequence (Genbank accession #AB592928.1). The coding sequence co-ordinates are: II sites . The glycassette to disrud RgBPst that alsGP55 ORF. The HCMV MIE promoter and polycloning linker sequence was isolated from pcDNA3 (Invitrogen) as a Bgl II/ EcoR V fragment and cloned into pACYC177 isolated as a BamH I/Nru I fragment to remove the kanamycin cassette to generate pACYCIE. The SV40 polyA cassette was then subcloned as an EcoR I fragment from an existing clone into pACYCIE cut with EcoR I to generate pACYCIESV. A full length gB expression construct was also generated by PCR cloning the complete ORF as a Bgl II fragment using forward primer Fgp55Bgl and reverse primer Rgp55fullBgl (First a low copy number expression plasmid was generated on the backbone of pACYC177 (New England Biolabs) to enable stable maintenance of the 5fullBgl . The fulGP75), gL (GP115) and gO (GP74) ORFs were additionally separately cloned into expression vectors that also tagged the C-terminal domain for easy detection of the recombinant protein in transfected cells. For GFP tagged gH, the GP75 ORF (missing the stop codon) was PCR amplified as a BamH I fragment using primers FgHBm and RgHBmNostop was PCR amplified as a Hind III fragment using primers FgLHd and RgLHdNostop as a EcoR V and Xho I fragment. This introduced an in frame 3x FLAG tag epitope tag into the C-terminal domain of gO and placed the gene under HCMV MIE promoter control. The modified plasmid was designated pGP74TAG8. A mutant version of the GP74 ORF was generated synthetically (DNA2.0) where all potential N-linked glycosylation sites (13 identified) were modified by changing the codon sequence of the putative N-linked glycosylation sites (NXT to NXA) to prevent glycosylation (see GP74 mutant ORF (designated GP74def) was placed under HCMV MIE promoter control in a mammalian expression plasmid pJ603 (DNA2.0 Inc.).For FLAG tagged gO, the hoNostop digestedtion see section.GP100) and gN (GP73) ORFs were separately cloned into expression vectors that also tagged the C-terminal domain . For GFP tagged gM, the GP100 ORF was PCR cloned as a BamH I (5\u2019) EcoR I (3\u2019) fragment lacking a stop codon into pAcGFP-N1 (Clontech) using primers gMTagF and gMTagR. For mCherry tagged gN, The GP73 ORF minus a stop codon was similarly PCR cloned as a BamH I (5\u2019) EcoR I (3\u2019) fragment into pmCherry-N1 (Clontech) using primers gNTagF and gNTagR. For FLAG tagged gN, both full length (codons 1\u2013132) and a N terminal truncated (codons 40\u2013132) GP73 ORF were PCR cloned separately (lacking a stop codon) into the expression vector pCMV3Tag8 (Agilent technologies Inc.) as BamH I / Hind III fragments to C-terminal epitope tag the ORFs using forward primers FGP73BmShort for truncated GP73 or FGP73BmFull for full length GP73 plus reverse primer RGP73nostopHd was introduced into DH10B bacterial cells containing a first or second generation GPCMV BAC plasmid , 21 usinmutation , 21. Insach gene . PCR reaHind III site encoded in the Km ORF. Modified GPCMV genomes were analyzed separately by EcoR I and Hind III restriction enzyme profile analysis. In an effort to limit redundancy the profiles shown for each mutant are either Hind III or EcoR I analysis. Additionally, two clonal mutants were generated for each knockout but only one is described. Comparative restriction fragment profiles of wild type and mutant GPCMV BAC genomes correctly demonstrated specific sub-genomic fragment modification for all mutants. Original designated GPCMV restriction fragment band nomenclature described by Gao and Isom [EcoR I profiles are shown for GP73, GP74, GP75, GP100 and GP115 knockouts. Hind III profile is shown for GP55 knockout. The GP73 ORF is encoded in the EcoR I \u2018C\u2019 fragment and was modified by a 1.1 kb Km marker insertion. This resulted in the modified \u2018C\u2019 fragment shifting and partially overlapping with the EcoR I \u2018B\u2019 fragment , see EcoR I \u2018C\u2019 fragment with a 1.1 kb shift in size because of the Km marker insertion, see EcoR I \u2018C\u2019 and \u2018B\u2019 fragments. An internal deletion within the ORF during mutagenesis , see EcoR I \u2018E\u2019 band and a 1.1kb insertion results in a shift in the band to 12.3 kb which partially overlaps with the \u2018D\u2019 fragment , see GP115 is encoded in the \u2018F\u2019 fragment and 1.1 kb insertion modifies the subgenomic fragment from 9,882bp to 10,982bp, see Hind III \u2018K\u2019 fragment was modified by Km cassette insertion which increased the size of the fragment by 700bp because of the presence of a Hind III site within the Km cassette, see GPCMV glycoproteins genes were individually knocked out by targeted mutagenesis of the GPCMV BAC in bacteria using shuttle vectors carrying a Km drug resistance marker to disrupt each ORF. The specific site of disruption for each glycoprotein is summarized in and Isom is used agenesis resultedThe modified loci for the GPCMV glycoprotein gene mutants were also confirmed by specific PCR analysis of wild type and disrupted glycoprotein genes using common flanking primers for each gene . The preRecombinant defective adenoviruses (serotype 5) encoding either GFP tagged gH or mCherry tagged gL or gB were generated as high titer stocks by Welgen Inc. on HEK293 cells. The C-terminal tagged ORFs from plasmids pAcGFPNgH and pmCherryNgL or the non-tagged complete ORF of gB from pACYCIEgB (described above) were each placed under HCMV MIE enhancer promoter control in the E1 locus of the defective Ad vectors using a IE1 shuttle vector (Welgen Inc.) to generate recombinant defective adenoviruses designated AdgHGFP, AdgLmCherry and AdgB respectively. A defective Ad vector encoding GFP (AdGFP) was also used in control expression studies.E. coli DH10B strain using a maxi plasmid kit (Qiagen). BAC DNA was transfected onto GPL cells in six well dishes using Lipofectamine 2000 (Invitrogen) as previously described [For generation of recombinant viruses, large-scale GPCMV BAC DNA was purified from escribed . GPCMV Bescribed . Non-infGP55, GP73, GP74, GP75, GP100 and GP115) were generated as previously described [trans. The GP75 ORF was placed under HCMV MIE promoter control in an existing sleeping beauty construct [For glycoprotein gene knockout mutants, each mutant was rescued back to wild type phenotype by co-transfection with the appropriate rescue plasmid. Rescue plasmids were generate as full length glycoprotein genes cloned individually into pUC19, pNEB193 (New England Biolabs) or pLITMUS28 (New England Biolabs). Rescue plasmids provided sufficient flanking homologous recombination sequence around the site of Km insertion on the mutated GPCMV BAC to enable efficient generation of rescue virus. All rescue viruses using primers previously described [Time point samples were taken from wild type GPCMV infected GPL cells in a six well dish (moi = 1 pfu/cell) at 0, 4, 8, 16, 24, 48 hr post infection. RT-PCR was performed essentially as described in McGregor et al. . RT-PCR escribed .Western blot and immunofluorescence assays were carried out as previously described , 51. HowAn in house anti-GPCMV IgG ELISA was carried out as previously described . BrieflyFor specific glycoprotein complex ELISA , GPL cells were transfected with expression plasmids encoding glycoprotein(s) or control GFP following previously described transfection protocol . As descAnti-GPCMV and anti-gB ELISA were performed on gB antibody depleted GPCMV convalescent pooled serum as described above. For anti-gB depletion, HEK 293 cells were transduced with recombinant defective adenovirus expressing gB (moi = 10 TDU/cell). Cells were harvested, washed twice with PBS, then fixed with 1:1 ratio acetone:methanol fixation mixture for 20 mins at -20\u00b0C. Fixed cells were pelleted then resuspended in 500\u03bcl of PBS + 1% tween 20. Equal volume of convalescent serum was used for preabsorption overnight at 4\u00b0C in a tube rotator. Cells were centrifuged at 10,000 x g for 20 mins at 4\u00b0C to pellet, serum collected then stored at -80\u00b0C until needed.Immunoprecipitation (IP) assays were carried out on plasmid transfected or recombinant Ad transduced fibroblast cells using commercial GFP-trap reagent (ChromoTek) following manufacturer\u2019s protocol and inclusion of protease inhibitor cocktail (Pierce) in cell lysates. Samples were subsequently analyzed by SDS-PAGE (4\u201320% gradient gel) and western blot using specific anti-epitope tag antibodies: HA ; FLAG ; GFP (Santa Cruz); Myc-c ; and mCherry (Clontech). Appropriate secondary anti-mouse or anti-rabbit HRP conjugate were also used following standard western blot protocol.GPCMV neutralization assays were conducted using convalescent GPCMV positive sera. Serially diluted guinea pig serum was incubated with approximately 50 pfu of GFP positive GPCMV, vAM403 , in mediGP55 (gB), GP73 (gN), GP74 (gO), GP75 (gH), GP100 (gM), GP115 (gL). Analysis of the predicted proteins indicated potential N-terminal signal leader peptide sequence of 23 and 24 amino acids in gB and gH respectively for gM (26 amino acids), gL (31 amino acids) and gN (44 amino acids), see GP74 is predicted to be heavily N-glycosylated as is the case for HCMV gO and the potential glycosylation sites are conserved between HCMV and GPCMV. This is discussed in more detail in a later section on the gH/gL/gO complex.Based on the GPCMV genome sequence , the GPCectively . N-termiGP55, GP75 and GP115 respectively) have been previously described.[GP73, GP74 and GP100 were analyzed by RT-PCR at various time points post infection (input virus moi = 1pfu/cell). The time course assay ; early, E, (GP54); and late, L, (GP83) [GP73, GP74 and GP100) are expressed as early/late genes. The use of transcription chemical inhibitors (cycloheximide and phosphonoacetic acid) more precisely place GP73 (gN), GP74 (gO) and GP100 (gM) as late transcripts , 58, 59.nscripts as transnscripts which deIn order to evaluate cellular location and interactions of the GPCMV glycoproteins, the various ORFs were cloned into commercial mammalian expression vectors and epitope tagged at the C-terminus. Construction of mammalian expression vectors encoding the GPCMV glycoprotein ORFs was as described in materials and methods. Transient expression studies were carried out on GPL cells and subsequently analyzed by: western blot, to verify protein size; immunofluorescence, for cellular co-localization of proteins; immunoprecipitation assay, to demonstrate specific protein:protein interactions.In HCMV, gM/gN proteins form a glycoprotein complex and poteIn contrast to gN, the gM (GP100) protein is not predicted to be as extensively glycosylated with two N-glycosylation sites predicted see . WesternCamelidae make a second antibody called heavy chain antibody that lack a light chain and bind their antigen via a single variable domain. This approach reduces potential background associated with the use of regular antibodies in IP assays. Manufacturer\u2019s protocol (ChromoTek) was followed to immunoprecipitate any proteins that interacted with gMGFP , see Fluorescence and immunofluorescence assays of transiently expressed GPCMV gH, gL and gO homolog proteins demonstrated that gH and gL co-localized in the cytoplasm of GPL fibroblast cells . ConsequA series of further transient co-expression studies were carried out in GPL cells with gHGFP and gO (both wild type and mutant versions) to evaluate the ability of gH to complex with gO in the absence of gL. The immune response to GPCMV and viral glycoprotein homolog complexes were investigated for pooled sera from animals convalescent for GPCMV. An in-house anti-GPCMV ELISA in comparison to the wild type locus was as predicted. The GPCMV mutant PCR products were additionally sequenced to further verify knockout mutations (data not shown).In order to determine the essential nature of the GPCMV glycoproteins, each gene was individually knocked out by targeted mutagenesis of the GPCMV BAC in bacteria using shuttle vectors carrying a kanamycin (Km) drug resistance marker to disrupt each ORF as described in materials and methods. The specific site of disruption for each glycoprotein is summarized in UL128-131 genes). All knockout GPCMV mutants could be successfully rescued by co-transfection of the appropriate rescue plasmid to generate wild type virus . Additionally, each mutant BAC was co-transfected with the appropriate rescue plasmid or rescue PCR product encoding the respective wild type glycoprotein gene to generate rescue virus from lethal knockout mutants see . The respe virus .The viable gO mutant virus was highly cell associated and produced virtually no detectable cell release virus. Viral spread on GPL cells was relatively restricted to primary plaques and was incapable of spreading across the full monolayer of cells. This was presumably due to the limited ability of the fibroblast cells to enable virus entry via endocytosis pathway. In epithelial cells, the gO mutant virus could successfully spread across the complete monolayer but had delayed growth kinetics compared to wild type virus . Given the lack of cell release virus produced by the gO mutant virus, it is likely that the gO protein has a role in viral maturation but confirmation awaits further study.This is the first report of a systematic knockout of the encoded glycoprotein genes of an animal cytomegalovirus and characterization of their glycoprotein complexes. The level of identity that exists between HCMV and GPCMV glycoproteins, as well as the conserved essential nature of viral proteins, implies that both HCMV and GPCMV glycoproteins have similar function in the viral life cycle. Since homolog glycoproteins to gB, gH, gL, gO, gM and gN are encoded by other animal CMV, it is probable that homolog glycoproteins from other species have a similarly conserved essential nature. In HCMV, variations in specific glycoprotein amino acid sequences have been identified in different clinical isolates of HCMV vs lab strains of HCMV. Two glycoproteins in particular (gN and gO) exhibit the greatest variation in sequence . A recenHCMV gO protein is extensively N-glycosylated with 18 potential of N-X-S/T sites and thisIn our GPCMV ELISA assays, we demonstrated that the antibody immune response in convalescent animals is directed to various glycoprotein complexes . In HCMVIn HCMV, both gH/gL and gB complexes are neutralizing antibody targets. Recently, the GPCMV gH/gL complex has also been shown to generate neutralizing antibodies in mice which woThe gM/gN complex in HCMV is important for virus entry and the essential nature of these proteins in GPCMV would also suggest a similarly important role. HCMV and GPCMV gM and gN proteins exhibit the highest identity of all the homolog proteins as well as conserved predicted multiple transmembrane domains which would suggest conservation of structure and function. The lack of specific antisera to GPCMV gM/gN precludes the direct demonstration of this protein complex role in cell entry. However, the GPCMV gM protein does have heparin binding capability (data not shown) which is a conserved feature of other gM proteins . In markOverall, the results from these studies suggest that GPCMV forms glycoprotein complexes similar to HCMV and that there is conserved function and properties in these complexes between HCMV and GPCMV. The complexes are important immunogenic targets in guinea pigs convalescent for GPCMV and importantly the neutralizing immune response is not solely directed to gB. Knockout mutagenesis of the individual viral glycoproteins confirmed their essential nature. A second method of entry into cells potentially exists in GPCMV with intact tropism to epithelial cells which enables gO independent infection of both fibroblasts and other cell types. However, our current data could also suggest that the pentameric complex can inefficiently perform the same function as the triplex via the same mechanism. A pending paper from our lab on epithelial tropism mutants would argue that the former is the case. Overall, these findings strengthen the guinea pig model for CMV and the continued development of intervention strategies based on this model.After more than fifty years of research, an effective intervention strategy against congenital cytomegalovirus remains an elusive goal. A complication with HCMV studies is the inability to directly utilize an animal model because of HCMV species specificity. GPCMV, a guinea pig specific virus, has emerged as highly relevant animal model because of the unique ability of the virus to cause congenital infection unlike mouse or rat models. An important aspect of CMV is the viral glycoproteins complexes that are present on the outside of the viral membrane. These are necessary for cell entry and are consequently important vaccine neutralizing target antigens. In this paper, we characterize the homolog glycoproteins and respective complexes in GPCMV and demonstrate conserved function between GPCMV and HCMV glycoproteins. Based on newly established assays for GPCMV, the GPCMV glycoprotein complexes are highly immunogenic. Additionally, manipulation of the GPCMV genome was carried out via herpesvirus BAC technology to generate specific mutants to characterize essential function of individual glycoprotein gene which showed similarity to HCMV. Overall, these studies are an important step in the continued use of this model for development of vaccine intervention strategies against CMV.S1 FigEcoR I (Ec) and Hind III (Hd) restriction profile analysis were performed for each mutant GPCMV BAC but only one profile is shown for each mutant to reduce repetition. Specific band shift are indicated as original wild type band (yellow) and modified mutant band (red). (i) Map of the GPCMV genome with individual glycoprotein genes indicated: GP55 (gB); GP73 (gN); GP74 (gO); GP75 (gH); GP100 (gM); GP115 (gL). Red indicates the gene is essential and green that the gene is semi-essential/non-essential for viable virus . Both (TIF)Click here for additional data file.S2 FigGP73), gO (GP74), gH (GP75), gM (GP100) and gL (GP115) the entire ORF was cloned and modified by kanamycin cassette insertion using indicated restriction sites. The GP75 ORF was modified by a collapse between two sites (Nru I and EcoR V). For the gB (GP55) the homolog AD-1 domain was PCR cloned as a shuttle vector with Km inserted into a unique EcoR V site as described in materials and methods to disrupt the ORF. The sizes of the original genes by PCR analysis are indicated and the sizes of the modified genes after kanamycin cassette insertion are also indicated Click here for additional data file.S3 FigGP74, GP75, GP100, GP115. (ii) GP73. (iii) GP55. GPCMV BAC mutant and wild type GPCMV analysis via PCR. Sample lanes: (1) GP74 wt; (2) GP74 mutant; (3) GP75 wt; (4) GP75 mutant; (5) GP100 wt; (6) GP100 mutant; (7) GP115 wt; (8) GP115 mutant (9) GP73 wt; (10) GP73 mutant; (11) GP55 wt; (12) GP55 mutant.Common primers were used to amplify the genes of wild type and mutant GPCMV. PCR primers as described in materials and methods and (TIF)Click here for additional data file.S4 Fighttp://www.cbs.dtu.dk/services/SignalP/ [http://sigpep.services.came.sbg.ac.at/signalblast.html. (E) gN leader sequence determined by http://www.csbio.sjtu.edu.cn/bioinf/Signal-3L/ [Various web based programs were used to predict the presence of a signal peptide sequence associated with individual proteins. (A) gB and (B) gH leader sequences determined by SignalP/ . (C) gM gnal-3L/ . Data sh(TIF)Click here for additional data file.S5 FigRT-PCR assays were carried out with GPCMV strain 22122 infected GPL cells in 6 well dish (moi = 1 pfu/cell) at different time points in the presence or absence of specific chemical inhibitors. Cycloheximide was used to prevent transcription of all but the IE transcripts and phosphonoacetic acid was used to prevent late transcripts as described in materials and methods. RT-PCR was carried out as described in materials and methods. Lanes: 1, bp ladder (Invitrogen); 2, mock infected; 3, 6 hour CHX treated; 4, 24 hour CHX treated; 5, 24 hour PAA treated; 6, 48 hour PAA treated; 7, no template control; 8, infected cell lysate no reverse transcriptase stage; 9, untreated (no inhibitor) GPCMV infected cell lysate. GP122 (IE2) RT-PCR is a positive control for GPCMV at specific assay time points treated with inhibitors. GAPDH is a positive cellular RNA control for all time point samples.(TIF)Click here for additional data file.S6 Fighttp://www.cbs.dtu.dk/services/TMHMM/). Potential transmembrane helices indicated in red in alignment with the predicted protein sequence .The predicted amino acid sequences for HCMV and GPCMV gM and gN proteins were analyzed for potential transmembrane domains by the web based program TMHMM Server v. 2.0 Prediction of transmembrane helices in proteins ((TIF)Click here for additional data file.S7 FigAnti-GPCMV sera depleted for anti-gB antibodies by preabsorption against Ad-gB transduced HEK 293 cells was verified for depletion by Western blot analysis as described in Materials and Methods. Lanes 1, 4, 7 mock infected GPL cells; Lanes 2, 5, 8 Ad-gB transduced GPL cell lysates (moi = 20 TDU/cell); Lanes 3, 6, 9 late stage GPCMV infected GPL cell lysates (moi = 1 pfu/cell). GPCMV convalescent sera (1:500) used for lanes 1\u20133, anti-gB depleted GPCMV sera (1:100) used for lanes 4\u20136. GPCMV gB monoclonal antibody (29\u201329) used for lanes 7\u20139 (1:500). Black arrow shows gB.(TIF)Click here for additional data file.S1 Table(DOC)Click here for additional data file."} +{"text": "Children and adolescents with systemic lupus erythematosis (SLE) are at risk for osteopenia and osteoporosis. Sex hormones protect against bone loss and as SLE itself and the immunosuppressive drugs given might interfere with normal puberty, this could add to the risk of bone loss among these patients.To evaluate the frequency of osteoporosis and hypogonadism among adolescents with SLE and how far the two conditions co-exist. The effect of disease characteristics and immunosuppressive drugs on both conditions are studied as well.Thirty-six adolescents with SLE were evaluated to determine the characteristics of the disease. SLE disease activity index (SLEDAI) was used to assess SLE status. Beside routine laboratory investigations of SLE, measurement of follicle stimulating hormone(FSH), leutinizing hormone (LH) and estrogen (E2) for females or testosterone for males before and 4 hours after LH releasing hormone analogue (0.1 ml) injection. Dual emission X-ray absorptiometery (DEXA) scan was done for all patients.Eighteen (50%) patients had low bone mass density(BMD), of these patients, 11 (31%) had osteoporosis (BMD < -2 Z-score) and 7(19%) had osteopenia (BMD <-1 and >-2 Z-score). Fifteen (42%) patients had hypogonadism: 12 (33%) had secondary hypogonadism and 3 (9%) had primary hypogonadism. The frequency of hypogonadism was comparable among patients with osteopenia/osteoporosis and those with normal BMD (p > 0.05). The age of the studied patients with osteopenia/osteoporosis had significant negative correlation with Z-score of DEXA scan (p < 0.05). Longer duration of SLE was associated with higher frequency of osteopenia/osteoporosis as well as hypogonadism among the studied patients (p = 0.01). While patients with osteopenia/osteoporosis had significantly higher SLEDAI as compared to those with normal BMD (p = 0.01), SLEDAI was comparable among patients with hypogonadism and those with normal gonadal function (p > 0.05). A significant negative correlation was found between the cumulative dose of steroids and Z-score of DEXA scan among patients with osteopenia and osteoporosis (p = 0.01). Patients with hypogonadism had received comparable cumulative doses of steroids and cyclophosphamide to those with normal gonadal function (p > 0.05).Osteopenia/osteoporosis and hypogonadism are common co-morbid conditions among adolescents with SLE. Lupus flare and steroids seem to affect BMD rather than gonadal function. Whether hypogonadism adversely affect BMD, this remains to be verified on a larger scale.None declared"} +{"text": "Induced pluripotent stem cells (iPSCs) derived from dilated cardiomyopathy (DCM) patients offer an unprecedented platform for in vitro disease modeling. Time coIn this study, dermal fibroblasts were isolated from skin biopsies of two unrelated patients who carry the RBM20 R636S mutation. The dermal fibroblasts were reprogrammed to iPSCs and then differentiated to cardiomyocytes to model the cardiogenesis for DCM patients. During the differentiation process, Cell samples at five stages were collected and RNA was extracted for time course transcriptome analysis. The iPSCs from a healthy subject was used as control.RBM20 familial DCM patient-specific cell lines and control showed hundreds of differential genes with 50 of them showing consistent differential expression patterns between the two disease cell lines. Gene function enrichment analysis performed on these 50 genes highlighted a vital functional group of pattern specification process including TBX18, CYP26B1, HHIP, and LHX2 (p \u2264 2.8E-8) which regulates cell response to differentiation during heart development.Unsupervised hierarchical clustering on genome-wide expression profiles defined clearly separated developmental stages containing pluripotent samples (day 0), early cardiac samples (day 10 and 15), and late cardiac samples (day 20 and 25). Furthermore, Principal Component Analysis revealed dramatic transcriptome differences on patients with severe and minor phenotypes. The comparison of transcriptome profiles of two RBM20 familial DCM due to dysfunctional cardiac gene expression in cardiogenesis. Insights gained from using patient-specific stem cells enables the anticipation of disease outcomes and targeting molecular therapy at the root cause of DCM.This study highlights developmental defects linked to the causative etiology of"} +{"text": "We read with great interest the recently published article \u201cExtraction of Iron from the Rabbit Anterior Chamber with Reverse Iontophoresis\u201d by Sun et al. . In thisFirst, as is known, ocular siderosis is caused generally by penetrating eye injury. Intraocular foreign body is most of the time located in vitreous cavity and scleral and/or corneal laceration is seen after penetrating eye injuries. Chiquet et al. reported that, after penetrating eye injury, IOFBs are commonly located in retina 23%), vitreous (48%), sclera (16%), and anterior segment (13%). The common ocular findings upon admission were corneal wound (66%), prolapse or damage of the iris (48%), hyphema (42%), and lens damage (42%) [3%, vitreOur experimental study showed that surgery should be performed within 14 days after penetrating eye injury with iron foreign body ; RI migh"} +{"text": "Calothrix strain 336/3, an N2-fixing heterocystous filamentous cyanobacterium isolated from a natural habitat. Calothrix 336/3 produces higher levels of hydrogen than Nostoc punctiforme PCC 73102 and Anabaena strain PCC 7120 and, therefore, is of interest for potential technological applications.We announce the draft genome sequence of The preCalothrix 336/3 is an N2-fixing heterocystous filamentous cyanobacterium isolated from the En\u00e4j\u00e4rvi lake, Laukilanlahti, Finland , using the Rapid Annotations based on Subsystem Technology (RAST) server , PCC 6303 (CO003610), and PCC 7103 (ALVJ00000000), respectively.The draft genome was automatically annotated with the NCBI Prokaryotic Annotation Pipeline (PGAP); server , 8 and t) server . The resCalothrix 336/3 genome contains nif (nifHDK1) and hup (hupLS) operons encoding nitrogenase and uptake hydrogenase enzymes but lacks hoxEFUYH genes encoding bidirectional hydrogenase and sets of nifHDK2 and vnfDGK genes encoding alternative nitrogenases, in line with the results obtained by Leino et al. with enzyme activity and Southern hybridization analyses (2 photoproduction capacity after immobilization in thin alginate films (Calothrix 336/3 genome opens new opportunities for potential technological applications in the development of biohydrogen production.Genome annotation revealed that the te films , 11. InvCalothrix strain 336/3 has been deposited at GenBank under the accession number JPKF00000000.The draft genome sequence of"} +{"text": "Arabidopsis thaliana and related plant species with similar characteristics as the prototype pattern, bacterial flagellin. Characteristic differences in flagellin and nlp20 plant responses exist however, as nlp20s fail to trigger extracellular alkalinization in Arabidopsis cell suspensions and seedling growth inhibition. Immunogenic nlp20 peptide motifs are frequently found in bacterial, oomycete and fungal NLPs. Such an unusually broad taxonomic distribution within three phylogenetic kingdoms is unprecedented among microbe-derived triggers of immune responses in either metazoans or plants. Our findings suggest that cytotoxic NLPs carrying immunogenic nlp20 motifs trigger PTI in two ways as typical patterns and by inflicting host cell damage. We further propose that conserved structures within a microbial virulence factor might have driven the emergence of a plant pattern recognition system mediating PTI. As this is reminiscent of the evolution of immune receptors mediating ETI, our findings support the idea that there is a continuum between PTI and ETI.Microbe- or host damage-derived patterns mediate activation of pattern-triggered immunity (PTI) in plants. Microbial virulence factor (effector)-triggered immunity (ETI) constitutes a second layer of plant protection against microbial attack. Various necrosis and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) produced by bacterial, oomycete and fungal microbes are phytotoxic virulence factors that exert immunogenic activities through phytotoxin-induced host cell damage. We here show that multiple cytotoxic NLPs also carry a pattern of 20 amino acid residues (nlp20) that triggers immunity-associated plant defenses and immunity to microbial infection in Eukaryotic host immunity to microbial infection requires recognition systems sensing the presence of potential invaders. Microbial surface structures (patterns) or host breakdown products generated during microbial attack serve as ligands for host immune receptors (pattern recognition receptors) mediating activation of immune responses. Microbial pathogens employ, however, host-targeting effector proteins to establish infection, and the efficiencies of microbial pathogen attack and host defense mechanisms determine the outcome of microbe-host interactions. Necrosis and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) from bacteria, oomycetes and fungi are cytotoxic virulence factors (effectors) that trigger plant immunity through toxin-induced host cell damage. Here we show that, in addition, numerous NLPs harbor a characteristic 20-mer sequence motif (nlp20) that is recognized by Brassicacae plant species and perception of which confers immunity to infection by bacterial, oomycete and fungal pathogens. Our findings provide evidence that cytotoxic NLPs are virulence factors that trigger plant immunity by pattern recognition and by inflicting host cell damage. We further conclude that NLPs from prokaryotic and eukaryotic microorganisms and from three organismal kingdoms evoke plant defense. Such an exceptionally wide taxonomic distribution of microbe-derived triggers of immunity has neither been reported before from metazoans nor from plants. Plants make use of a bipartite immune system to cope with microbial infection Sclerotinia sclerotiorum-derived proteinaceous SSCF1 or Xanthomonas campestris-derived EMAX are recognized by Brassicaceae only Arabidopsis thaliana ecotypes have further revealed that individual pattern recognition specificities might also be lost during evolution Plants recognize a wide range of proteinaceous, carbohydrate or lipophilic PAMPs Plant pathogenic microbes produce multiple effector proteins that are secreted into the plant apoplastic space or that are translocated into host cells by means of specialized translocation systems, such as type III secretion systems of Gram-negative bacteria Pythium aphanidermatum or Moniliophthora perniciosa NLPs, respectively, revealed substantial fold conservation with cytolytic, pore-forming actinoporins from marine organisms, suggesting that NLPs destabilize plant plasma membranes during infection thereby facilitating host cell death Pectobacterium carotovorum pv. carotovorum (PccNLP), Pythium aphanidermatum (PyaNLP) or Phytophthora parasitica (PpNLP) were shown to be key virulence factors sharing identical fold requirements for NLP phytotoxin and virulence activities NLPs form a superfamily of proteins that are produced and secreted by bacterial, fungal and oomycete species Mycosphaerella graminicola produces MgNLP that is toxic on dicot plants, but not on its monocot host, wheat Verticillium dahliae resulted not only in reduced virulence on host plants, but also in reduced vegetative growth and conidiospore formation, suggesting a role of this NLP in asexual reproduction in addition to its role in fungal pathogenicity Hyaloperonospora arabidopsidis was shown to produce up to 10 NLP proteins all of which failed to cause necrosis in dicot plants including the host ArabidopsisPhytophthora sojae NLPs tested lacked phytotoxic activities There is accumulating evidence that NLP effectors have diversified in function PccNLP inactive with respect to cytotoxicity, host virulence and plant immune activation, also abolished the cytotoxic activity of another NLP (PpNLP), but surprisingly left intact its ability to trigger plant defenses. This suggested the presence of another, yet unidentified immunogenic activity of PpNLP. The elicitor activity of mutated PpNLP could be pinpointed to a peptide fragment (nlp20) that triggered plant defenses in a manner comparable to that of bacterial flagellin. Importantly, immunogenic nlp20 fragments were found frequently in NLPs of bacterial, oomycete and fungal origin. In sum, we demonstrate that a common microbial effector harbors a PAMP motif that is found in both prokaryotic and eukaryotic microbes. Thus, its widespread occurrence is unique among microbial triggers of metazoan or plant innate immunity. In addition, the identification of two independent plant immunogenic mechanisms (PAMP- and toxin-induced immunity) within a particular microbial virulence factor is unprecedented and reveals an intricate complexity of microbial virulence and plant immune activation.In this study, we have investigated plant immunogenic activities of NLP virulence factors in greater detail. Mutations that rendered Pectobacterium carotovorum pv. carotovorum-derived PccNLP and Phytophthora parasitica-derived PpNLP cause necrosis upon infiltration into leaves of Arabidopsis thaliana (PpNLP) abolished necrotic did not affect plant defense-eliciting activity proved both to be able to elicit plant defense-associated responses. These peptides share residues G100-D113 or C-terminal deletion of residues K112 and D113 (peptide 2) substantially reduced elicitor activity, suggesting that both motifs are important for the immunogenic potential of PpNLP (112D113 (peptide 2) or peptides with C-terminal extensions, but lacking residues Y102-Y106 were all inactive. Substantial N-terminal extension (peptide 8) did not increase elicitor activity of this peptide in comparison to peptide 1, suggesting that no further sequence information N-terminal of the G100-Y106 motif is required for elicitor activity of PpNLP. To refine C-terminal sequence requirements for PpNLP elicitor activity, we further tested peptides containing the Y102-Y106 motif or a fragment thereof (A103-Y106) and different C-terminal extensions beyond residues K112D113 or 1,5 nM (peptide 13), respectively, as the most elicitor-active peptides, which are both substantially more active than peptide 1 and nlp24 (PpNLP), respectively. To identify amino acids within both peptides that are essential for their elicitor activities, an alanine-scanning mutagenesis was conducted (103W) identified residues I104, Y106, W108, and Y109, of which replacement reduced immunogenic activities of mutant peptides more than 1,000-fold as compared to nlp24 (PpNLP). All other exchanges had significantly less or no effect on the activities of the mutant peptides or nlp24 (PpNLP), respectively, affected immunogenic activities of the mutant peptides in a rather moderate manner.Typically, small epitopes within microbial patterns are sufficient for their immunogenic activities pression . Two oves 322 nM . N-termiof PpNLP . In agreK112D113 . These septide 1 . As bothonducted . Individpeptides . ImportaPpNLP) motif derived from cytotoxic NLP would retain both immunogenic and cell death-causing activities, leaf necrosis and plasma membrane permeabilization assays were performed using equimolar concentrations of intact PpNLP and of PpNLP-derived nlp20 (PpNLP) as well as 10-fold higher concentrations of the latter. As shown in PpNLP) failed to trigger either response, suggesting strongly that its immunogenic activity is not linked to cell death or plasma membrane disintegration. This conclusion is further supported by our findings that heat treatment or mutations within intact PpNLP abolished its necrosis-inducing activity, but not its ability to trigger immunity-associated defenses (PpNLP) are required to trigger plant defenses (PpNLP)-induced defenses from the cytotoxic potential of intact PpNLP.To test whether the nlp20 , fungus- or oomycete-derived (Pythium aphanidermatum) sequences orthologous to nlp20 (PpNLP) were analyzed for their immunogenic potential. As shown in PR1::GUS expression, and callose apposition or orthologous nlp20 peptides were tested , proved largely inactive with respect to activating ethylene formation, MAPK activation and PR1::GUS expression (50 5520 nM), which was approximately 400 times less active than nlp20 (PpNLP) (EC50 14 nM) (As shown in pression . Residua0 14 nM) .Brassicaceae species beside Arabidopsis thaliana responded to this peptide. As shown in Arabis alpina, Thlaspi arvense, and Draba rigida mounted an ethylene response to nlp20 (PpNLP) treatment, suggesting that nlp20 recognition is widespread among the Brassicaceae family. Notably, another species from the genus Arabidopsis, Arabidopsis lyrata, did not respond to nlp20 (PpNLP), but did so to the control treatment with flg22. Although surprising in the first place, this finding might just reflect that PAMP responsiveness is often even not entirely conserved among ecotypes of the same species. Neither solanaceous plants nor parsley or wheat responded to nlp20 (PpNLP) (PpNLP) responses in parsley is in agreement with our previous studies showing that this plant species lacks the ability to recognize NLP peptide fragments Asteraceae family) with nlp20 (PpNLP) (PpNLP) derivative lacking PAMP activity in Arabidopsis thaliana (PpNLP) perception systems in both plants exhibit similar ligand specificities. Whether nlp20 recognition is even more widespread among plant families requires comprehensive, systematic surveys of its immunogenic activity.To analyze the relative distribution of nlp20 recognition systems among plants, we first tested whether other (PpNLP) . Failure (PpNLP) . As lettthaliana we conclArabidopsis plants prior to infection with virulent Pseudomonas syringae pv. tomato DC3000 reduced bacterial growth by about 100-fold within three days post infection when compared to bacterial growth rates on mock-treated plants (PpNLP) treatment limited bacterial growth rates on ecotype Col-0 to a similar extent as did flg22 treatment, suggesting that both patterns have an immunogenic activity (PpNLP) also reduced bacterial growth on an fls2 efr genotype (PccNLP) (PpNLP) treatment also primed Arabidopsis plants for enhanced immunity to infection by the fungal phytopathogen Botrytis cinerea. Lesion sizes in plants pretreated were significantly smaller than those observed in mock-treated plants. Likewise, pre-treatment of lettuce with an nlp24 peptide derived from H. arabidopsidis nlp24 (HaNLP3) enhanced resistance to infection with Bremia lactucae or pepti(PccNLP) did not (PccNLP) . As furtlactucae . AltogetPccNLP Fusarium spp.-derived fumonisin, Phomopsis amygdali-derived fusicoccin or Cochliobolus victoriae-derived, victorin. Toxin-induced immunity is thus considered a hallmark of innate immunity not only in metazoans, but also in plants Cytotoxic NLPs are microbial virulence factors facilitating both microbial infection and activation of plant immunity-associated responses. Toxin-mediated release of diffusible DAMPs from lyzed plant cells and subsequent PRR-mediated plant immune activation in neighboring cell layers or local systemic tissues has been proposed as the likely molecular mechanism underlying immunogenic activity of, for example, PccNLP non-cytotoxic and non-immunogenic failed to have the same effect in other cytotoxic NLPs, such as PpNLP. We have now been able to identify a peptide motif (nlp20 motif) within PpNLP and other NLPs that is missing in PccNLP. This strongly suggests that cytotoxic NLPs carrying the nlp20 motif are potentially capable of evoking plant immunity by two different mechanistic modes, by toxin action and by a classical PAMP motif. To our knowledge, this is an unprecendented finding as microbial patterns with dual immunogenic activities are currently unknown in both metazoan and plant immunity. These results shed light on how intricately complex and mechanistically diverse microbe sensing in individual plant microbe encounters might be. In support of this notion, Arabidopsis thaliana alone is capable of recognizing at least seven structurally different patterns derived of pseudomonads To our surprise, we have been able to unveil a second molecular mechanism by which cytotoxic NLPs are able to evoke plant immunity. This discovery was spurred by findings that mutations that rendered PpNLP1 sequence as query . Preliminary inspection of these sequences for the presence of the nlp20 motif and of those residues that are crucial for its PAMP activity revealed that a remarkably low number of bacterial sequences (20 out of 221), but a majority of fungal and virtually all oomycete NLPs likely contain an elicitor-active nlp20 motif. In sum, this motif is a predominant feature within a vast number of NLP sequences particularly in eukaryotic NLP-producing microorganisms. Importantly, in comparison to the relatively small numbers of NLP-encoding genes in fungal genomes, the number of NLP genes has expanded significantly in oomycete species. For example, the P. sojae genome harbors 33 NLP genes 20 of which have been shown to be expressed, whereas H. arabidopsidis encodes 12 NLP genes 8 of which are expressed early during plant infection P. carotovorum) or whether nlp20 motif-containing NLPs have been acquired from eukaryotic species via horizontal gene transfer are difficult to make as of now.PAMPs triggering immunity in metazoans or plants are supposed to be widespread among microbial species Arabidopsis cell suspensions and Arabidopsis seedling growth retardation The nlp20 motif exhibits molecular features similar to that of the prototype immunogenic pattern, bacterial flagellin (flg22) In summary, we here report the identification of a common immunogenic pattern within a microbial virulence factor. The nlp20 motif of bacterial, fungal or oomycete NLPs possesses the ability to trigger plant immune responses in a manner comparable to bacterial flagellin. Unique features of this pattern comprise (i) its presence in both prokaryotic and eukaryotic microbes and (ii) the fact that it constitutes a second immunogenic principle within cytotoxic NLPs. Further, we suggest that a microbial effector might have driven the emergence of plant pattern recognition systems mediating PTI. This is important as it is reminiscent of the evolution of immune receptors mediating recognition of pathogen race-specific microbial effectors and activation of ETI Arabidopsis Col-0 and efr fls2 plants were grown in soil at 22\u00b0C, 8 h light and used for the experiments at an age of 5\u20136 weeks. Plants used for infection assays were grown under translucent cover.Arabidopsis thaliana Col-0 plants were primed 24 hours before bacterial or fungal infection by leaf infiltration of nlp20 (PpNLP), flg22, C6 (1 \u00b5M peptide solution) or mock-treatment, respectively. To assess bacterial growth rates, Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) strain was used. The strain was maintained at 28\u00b0C on King's B medium containing rifampicin and cycloheximide (50 \u00b5g ml\u22121). Overnight cultures were centrifuged, washed twice in 10 mM MgCl2 and adjusted to a bacterial density of 104 cfu ml\u22121. Primed leaves were pressure-infiltrated with the bacterial solution and the plants were kept under high humidity. Leaves were harvested and surface sterilized in 70% EtOH and ddH20 for 1 minute each. Two leaf discs per plant were stamped out, ground in 10 mM MgCl2, diluted serially 1\u223610 and plated on LB plates containing the appropriate antibiotics. After 2 days of incubation, colony-forming units were counted. For fungal infection, primed Arabidopsis leaves were drop-inoculated with 5 \u00b5l droplets of Botrytis cinerea isolate BO-10 containing 5\u00d7106 spores ml\u22121 in PDB and kept under high humidity. Photographs were taken 2 days after infection and lesion sizes were determined using the Photoshop CS6 Lasso tool. Selected pixels were counted and the lesion size in cm2 was calculated using a 0,5 cm2 standard. For oomycete Bremia lactucae infection, L. sativa leaf discs were vacuum-infiltrated with 1 \u00b5M nlp24 (HaNLP3) and 24 hours later treated with a 20 \u00b5l droplet spore suspension (120 spores/\u00b5l). Sporulation was assessed 8 days post inoculation.5\u20136 weeks old Pichia pastoris GS115 or in the NLP-deficient Pectobacterium carotovorum subsp. carotovorum SCC3200 strain (\u2212Pcc nlp). Isolation of PccNLP proteins from the periplasmic space of transgenic \u2212Pcc nlp was performed by osmotic shock as described PpNLPs from P. pastoris culture medium or from P. carotovorum subsp. carotovorum SCC3200 periplasmic protein solution was achieved by ion exchange chromatography followed by gel filtration . As ion exchanger either HiTrap Q FF (equilibrated in 20 mM Tris-HCl pH 8.5: PpNLP) or HiTrap SP FF (equilibrated in 50 mM MES pH 5.7: PccNLP) was used. Following elution (0\u2013500 mM KCl in equilibration buffer), NLP containing fractions were pooled and subjected to HiLoad\u2122 16/60 Superdex 75, equilibrated in 150 mM KCl in the corresponding buffer. NLP containing fractions were finally pooled and dialyzed against H2O. Protein concentrations were calculated by UV spectroscopy (wavelength \u03bb280) using the protparam tool (http://web.expasy.org/protparam) to determine protein-specific extinction coefficients \u03b5280 for each protein. Determinations were verified by SDS-PAGE using a standard protein solution.For functional studies, secretory expression of NLPs was performed either in Peptides were purchased from Genscript Inc., prepared as 10 mM stock solutions in 100% DMSO, and diluted in water prior to use. DMSO concentrations corresponding to those in peptide solutions used in this study did not trigger themselves any of the responses shown here.2O overnight, were placed in one well of a 96-well plate, containing 100 \u00b5l of a 20 \u00b5M L-012 and 0.5 \u00b5g ml\u22121 peroxidase solution. Background was measured shortly in a 96-well Luminometer, before elicitation with a peptide solution or control treatment respectively. The detection of ethylene was performed as described Arabidopsis cells and detection of GUS enzyme activity in PR1::GUS transgenic Arabidopsis plants were performed as described previously Arabidopsis Col-0 seeds were grown in \u00bd MS liquid medium supplemented with 1 \u00b5M of nlp20 (PpNLP) peptide or its orthologs respectively, and flg22 or H2O serving as controls. Root length of two weeks-old seedlings was determined upon transfer onto agar plates.For MAPK activity assays, infiltrated plant material was harvested after 15 minutes and frozen in liquid nitrogen before used for protein extraction in 20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0,1% SDS, 5 mM DTT, Complete Protease Inhibitor Mini, EDTA-free , PhosStop Phosphatase Inhibitor Cocktail . After pelleting the cell debris , the supernatant (30 \u00b5g protein) was separated on a 10% SDS-PAGE and transferred to a nitrocellulose membrane and activated MAPK6, 3 and 4 were detected by western blotting using the anti phospho p44/42-MAPK antibody from rabbit . For ROS burst measurements two leaf pieces, floated on ddHArabidopsis leaves were infiltrated with 300 nM PpNLP or PccNLP, heat-denatured proteins or mutant versions (H121A D124A), respectively. RNA was isolated using the RNeasy Plant MiniKit (Qiagen) and synthesis of cDNA was performed by means of the RevertAidTM MuLV reverse transcriptase (Fermentas). Quantitative real-time PCR amplification was carried out in the presence of SYBR Green (Bio-Rad) with an iQ5 iCycler (Bio-Rad). Amplification of EF1-\u03b1 served as internal standard. Data were analyzed according to the 2\u2212\u0394\u0394CT-method 2O infiltration.Arabidopsis plasma membrane vesicles was performed as described Calcein release from intact Figure S1Comparison of cytotoxic and immunogenic activities of PpNLP, PccNLP and nlp20 (PpNLP). Calcein release induced by PpNLP, PccNLP or nlp20 (PpNLP) from purified plasma membrane vesicles prepared from Arabidopsis thaliana leaves. Vesicles were treated with either 333 nM wild-type NLP, heat-treated NLP (95\u00b0C), mutant (mut) NLP or nlp20 (PpNLP) peptide. Calcein release is calculated as the percentage of the maximum release as determined by addition of Triton X-100 at the end of the assay (A). Ethylene formation triggered upon Arabidopsis leaf infiltration of different concentrations of wild-type, heat-treated (95\u00b0C), mutant (mut) NLP variants (B). Data points represent n\u200a=\u200a3 repetitions, and one representative experiment of three is always shown. Image of a Coomassie-stained SDA-PAGE gel documenting the purity of recombinant proteins used (C). Leaf necrosis (D) and calcein release (E) triggered by nlp20 (PpNLP) peptide. One representative experiment of three is shown (C\u2013E).(EPS)Click here for additional data file.Figure S2Immunity-associated responses in Arabidopsis elicited by nlp20 (PpNLP) and its orthologs. Ethylene formation (A), MAPK activation (B), production of reactive oxygen species (C), PR1::GUS expression (D) and callose apposition (E) were determined upon infiltration into leaves of 100 nM nlp20Pp orthologous peptides derived from microorganism given in A) and (C) share the same color code. In (E), the diagram depicts callose apposition in % \u00b1 SD of three image sections of the leaf surface, counted as pixels. Photographs show the microscopic images after clearing callose depositions from background and leaf-veins. Arabidopsis seedlings were grown for two weeks under short day conditions in liquid \u00bd MS medium supplemented with 1 \u00b5M nlp20Pp peptide, its orthologs, or flg22 as a positive control respectively, and root length was determined after transfer onto agar plates. The upper panel documents quantification of 3 representative seedlings shown in the lower panel (F). Arabidopsis cell suspensions were supplemented with the nlp20Pp concentrations indicated or 100 nM flg22 and changes in extracellular pH were monitored continuously (G). All assays were performed in triplicate with similar results using the protocols described in (EPS)Click here for additional data file.Figure S3Nlp20 (PccNLP) does not induce defense responses in Arabidopsis. Elicitation of ethylene formation (A), MAPK activation (B) and PR1::GUS expression (C) in leaves infiltrated with peptide concentrations as indicated (A) or 100 nM . Peptides used were nlp20 (PpNLP) (closed circles) and nlp20 (PccNLP) . (B) Activation of MAPK6, 3 and 4 detected 15 minutes after leaf infiltration as visualized detected by anti p44/p42 antibody staining. Ponceau S staining served as a loading control. (C) For PR1::GUS expression analysis, leaves were harvested 24 hours after treatment and stained histochemically. One of three experiments is shown.(EPS)Click here for additional data file.Figure S4Lactuca sativa recognizes nlp20 (PpNLP). Leaf pieces of Lactuca sativa were infiltrated with 1 \u00b5M nlp20 (PpNLP) or an inactive variant Click here for additional data file.Figure S5Treatment with nlp20 (PpNLP) renders Arabidopsis efr fls2 less susceptible to bacterial infection (A), but elicitor-inactive nlp20 derivatives fail to prime plants for immunity to subsequent infection (B). Leaves were infiltrated with 1 \u00b5M synthetic nlp20 (PpNLP) (A), nlp20 (PccNLP) or peptide 20 (see B) 24 hours before inoculation of the same leaf with 104 cfu ml\u22121Pseudomonas syringae pv tomato strain DC3000 (Pst DC3000). Bacterial growth was determined at 0 and 3 days after leaf infiltration. Flg22 and water served as positive and negative controls respectively. Data represent means \u00b1 SD of six replicate measurements per treatment and data point. Asterisks indicate statistically significant differences to water control treatments . One of three independent experiments is shown.e 20 see (B) 24 h(EPS)Click here for additional data file.Figure S6Treatment with Hyaloperonospora arabidopsidis nlp24 (HaNLP3) renders Lactuca sativa less susceptible to the oomycete, Bremia lactucae.L. sativa cv. Olof leaf discs were vacuum-infiltrated with 1 \u00b5M nlp24 (HaNLP3) 24 hrs prior to inoculation with 20 \u00b5l of a B. lactucae isolate Bl:24 spore suspension (120 spores/\u00b5l). Oomycete sporulation was assessed 8 days post inoculation. Asterisks indicate statistically significant differences to water control treatments ***P\u22640.001, Student's t test). Experiments were repeated in triplicate with similar results.(EPS)Click here for additional data file."} +{"text": "Pneumothorax, though rare, is a recognized cause of respiratory distress in the immediate newborn period. It may occur spontaneously or secondary to various underlying lung diseases. Here we share our experience of a neonate with spontaneous pneumothorax with mild to moderate respiratory distress, who recovered completely with conservative management with an oxygen-enriched atmosphere and no surgical intervention. A 3350 g female neonate was born at term gestation by caesarean section to a primigravida mother after an uneventful antenatal period. There was no history of trauma during delivery or meconium stained liquor. She cried immediately after birth and did not require any resuscitation. At 30 min of life, she developed tachypnea with grunting and cyanosis (SpO2 varying from 80 to 83%). Supplemental oxygen via hood (FiO2 40%) was started after which cyanosis resolved (SpO2 increased to 88-90%), but respiratory distress persisted. Physical examination revealed a heart rate of 156/min, capillary refill time of 3 seconds, good volume pulses, and respiratory rate of 68/min. Breath sounds were decreased bilaterally without any crepitations or wheeze. Rest of the systemic examination was within normal limits. A chest x-ray was ordered. Arterial blood gas measurement (while the neonate was on 40% FiO2) showed combined respiratory with metabolic acidosis without hypoxemia . Meanwhile, the chest radiograph revealed bilateral pneumothoraces with normal pulmonary vasculature and normal cardiac silhouette . Sepsis screen , and blood culture were negative.The oxygen flow increased to 100% (via oxygen hood). Oro-gastric tube feeding with mother\u2019s milk was continued along with continuous monitoring of the clinical condition. Respiratory distress decreased within 24 hours. Gradually the FiO2 was decreased, and she was put off the oxygen after 72 hours. She did not require any surgical intervention in form of needle thoracocentesis or chest tube placement. Repeat chest radiograph done after 24 hours showed resolution of pneumothoraces .Pneumothorax can cause neonatal respiratory distress. It can occur spontaneously or secondary to respiratory distress syndrome, aspiration of meconium, etc. The incidence of pneumothorax is 1 to 2% in term newborns. It increased to about 6% in premature babies because of poor lung compliance.[3] In one study the frequency of pneumothorax was found to be 3 per 1000 live births.[4]Spontaneous pneumothorax at birth results either from rupture of alveoli secondary to high pressure needed to expand previously uninflated lungs or from uneven distribution of inflating pressures among alveoli. In familial cases of spontaneous pneumothorax, folliculin gene disorders or \u03b11-antitrypsin deficiency should be ruled out.[5] Sometimes cystic fibrosis may present with bilateral spontaneous pneumothorax during newborn period.[6]Administration of high flow or 100% oxygen (nitrogen washout therapy) accelerates the resolution of pneumothorax.[7] To conclude, all cases of pneumothorax in newborns do not require intercostal drain. Newborn with spontaneous pneumothorax having mild or moderate distress may recover completely with no treatment other than observation in an oxygen-enriched atmosphere. For prolonged treatment (> 48 hours) with oxygen, a lower FiO2 (40%-60%) can be used safely without causing much harm.Source of Support: NilConflict of Interest: None declared"} +{"text": "Dusp1 (DUSP1 knockout mouse).Osteoarthritis (OA) is a degenerative joint disease with poorly understood etiology and pathobiology. Mitogen activated protein kinases (MAPKs) including ERK and p38 play important roles in the mediation of downstream pathways involved in cartilage degenerative processes. Dual specificity phosphatase 1 (DUSP1) dephosphorylates the threonine/serine and tyrosine sites on ERK and p38, causing deactivation of downstream signalling. In this study we examined the role of DUSP1 in spontaneous OA development at 21 months of age using a genetically modified mouse model deficient in Utilizing histochemical stains of paraffin embedded knee joint sections in DUSP1 knockout and wild type female and male mice, we showed similar structural progression of cartilage degeneration associated with OA at 21 months of age. A semi-quantitative cartilage degeneration scoring system also demonstrated similar scores in the various aspects of the knee joint articular cartilage in DUSP1 knockout and control mice. Examination of overall articular cartilage thickness in the knee joint demonstrated similar results between DUSP1 knockout and wild type mice. Immunostaining for cartilage neoepitopes DIPEN, TEGE and C1,2C was similar in the cartilage lesion sites and chondrocyte pericellular matrix of both experimental groups. Likewise, immunostaining for phosphoERK and MMP13 showed similar intensity and localization between groups. SOX9 immunostaining demonstrated a decreased number of positive cells in DUSP1 knockout mice, with correspondingly decreased staining intensity. Analysis of animal walking patterns (gait) did not show a discernable difference between groups.Loss of DUSP1 does not cause changes in cartilage degeneration and gait in a mouse model of spontaneous OA at 21 months of age. Altered staining was observed in SOX9 immunostaining which may prove promising for future studies examining the role of DUSPs in cartilage and OA, as well as models of post-traumatic OA. Osteoarthritis (OA) is a degenerative joint disease which is estimated to afflict at least 10% of the US population over the age of 25 .At the tissue level, chondrocytes are the only active cellular component of the cartilage that caps the bone in articular joints such as the knee, elbow and ankle. These cells maintain tissue homeostasis by balancing anabolic buildup and catabolic turnover of surrounding extracellular matrix (ECM) proteins . The ECMin vitro, and inhibition of EGFR in vivo partially protects against surgically induced osteoarthritis in rats , TEGE, and C1,2C with appropriate rabbit normal IgG control and no primary added controls. (B) Immunostaining for MMP13 and phosphorylated ERK (phERK) with rabbit normal IgG control and no primary added controls. (C) Immunostaining for SOX9 with goat normal IgG and no primary added controls. Scale bars = 100 \u03bcm. Representative images shown. N\u22653.Female frontal knee sections were immunostained for (TIF)Click here for additional data file.S4 FigDusp1 KO mice show decreased numbers of SOX9 positive cells relative to WT controls. Data is presented as individual data points with mean \u00b1 SEM. Data analyzed using Mann-Whitney test.The number of SOX9 positive cells within a 200 x 100 \u03bcm box set at the articular cartilage surface of the lateral tibial plateau were counted and normalized to the animal\u2019s weight to correct for any variability caused by differences in joint loading and animal size. (TIF)Click here for additional data file.S1 TableDusp1-/-), Het (Dusp1+/-), KO (Dusp1-/-) animals live and healthy (Live) at the end of the experiment, or removed from the experiment due to recurrent skin ulcerations/died of unknown causes (Dead/Rem.).DUSP1 WT ((DOCX)Click here for additional data file.S2 TableArticular cartilage damage was evaluated using the OARSI recommended scoring system. SEM, standard error of the mean; WT, wild type; KO, knockout; LTP, lateral tibial plateau; LFC, lateral femoral condyle; MTP, medial tibial plateau; MFC, medial femoral condyle.(DOCX)Click here for additional data file.S3 TableArticular cartilage thickness was measured from the cartilage surface to the subchondral bone. SEM, standard error of the mean; \u03bcm, micrometers; WT, wild type; KO, knockout; LTP, lateral tibial plateau; LFC, lateral femoral condyle; MTP, medial tibial plateau; MFC, medial femoral condyle.(DOCX)Click here for additional data file."} +{"text": "We present a case report of a patient with glioblastoma multiforme (GBM) complicated by extracranial metastasis (ECM) whose survival of nearly four years surpassed the anticipated life expectancy given numerous negative prognostic factors including EGFRvIII-mutation, unmethylated MGMT promoter status, and ECM. Interestingly, while this patient suffered from locally aggressive disease with multiple intracranial recurrences, the proximal cause of death was progressive extracranial disease and complications related to pulmonary metastases. Herein, we review potential mechanisms of ECM with an emphasis upon glioblastoma molecular and genetic profiles and the potential implications of targeted agents such as bevacizumab. Glioblastoma multiforme (GBM) is one of the most lethal human malignancies. The median survival of patients with newly diagnosed GBM treated with adjuvant radiation therapy (RT) with concurrent and adjuvant temozolomide (TMZ) remains dismal at 14.6 months. The majority of patients ultimately succumb to local recurrence in the central nervous system , 3. HoweCurrent literature suggests that the median time from initial diagnosis to detection of ECM is approximately 8.5 months and the interval of time between detection of distant metastasis and death is only 1.5 months, suggesting that development of ECM is a harbinger of morbidity and mortality , 3. The A previously healthy 51-year-old right-handed African American gentleman initially presented following an unwitnessed seizure. MRI revealed focal enhancement in the posterior temporal lobe measuring approximately 9\u2009mm as well as T2 hyperintensity within the right insular cortex and right temporal lobe. He was empirically treated for HSV encephalitis until CSF HSV PCR assay returned negative. Given persistent mental status changes, MR imaging at 4 and 9 months after initial presentation revealed an enhancing temporoparietal mass with interval enlargement between scans. He underwent right temporal craniotomy with postoperative pathology consistent with WHO grade IV astrocytoma (GBM) with small cell features . Specifi2/day) followed by adjuvant TMZ (150\u2009mg/m2/day \u00d7 5 days every 28 days). Follow-up MRI after two cycles of adjuvant TMZ revealed increased enhancement within the surgical cavity. TMZ was switched to metronomic scheduling (120\u2009mg daily) during the third cycle due to concern of progression versus pseudoprogression for a total of 8 additional cycles. MR imaging a month later revealed marked progressive enhancement and the patient underwent repeat resection with postoperative imaging indicating gross total resection and pathologic confirmation of recurrent GBM (He was treated with adjuvant radiotherapy (RT) delivered (6000\u2009cGy in 30 fractions) with concurrent daily temozolomide .At 27 months following his initial diagnosis, the patient initiated bevacizumab monotherapy which was discontinued after four cycles due to radiographic progression with development of a mildly enhancing intracavitary nodule. A third craniotomy was performed with pathology demonstrating mitotically active, recurrent GBM . The patAfter two months of therapy in the aforementioned clinical trial he developed left homonymous hemianopia with brain MRI revealing infiltrative enhancement around the right temporal lobe surgical cavity and increased vasogenic edema throughout the right temporal and parietal lobes. He underwent reirradiation (3600\u2009cGy in 20 fractions) with concurrent bevacizumab. He continued with adjuvant bevacizumab for an additional two cycles and maintained a good performance status throughout therapy.The patient developed back pain and abdominal discomfort approximately 40 months following initial diagnosis and 3 months following reinitiation of bevacizumab therapy. CT imaging of the chest demonstrated multiple pulmonary nodules, including a 2.5\u2009cm right middle lobe nodule and a 1.2\u2009cm left lower lobe nodule along with bilateral hilar adenopathy. A staging fluorodeoxyglucose positron emission tomography (FDG PET) scan noted abnormal uptake within several right hilar lymph nodes and diffuse osseous FDG uptake within the sternum, thoracic, lumbar, and pelvic bones. The pulmonary nodules identified on CT imaging were also found to be FDG-avid, right middle lobe (SUV 11.5), two left upper lobe nodules (SUV 6.4 and 5.1), and right upper lobe (SUV 5.0) . FlexiblOne month later, he resumed treatment with bevacizumab and, in the 43rd month, carboplatin. Restaging FDG PET imaging after 2 cycles revealed extracranial progression of disease with worsening pulmonary and skeletal metastases. Of note, synchronous brain MRI was stable at this time with no evidence of intracranial recurrence or progression . He contIt is currently unknown if the use of bevacizumab is implicated in the potentiation of ECM. It is notable that extracranial dissemination of this patient's disease coincided with administration of bevacizumab therapy.Concerns have been raised that anti-VEGF therapies, including bevacizumab, lead to transformation of primary and recurrent GBM into a more infiltrative phenotype , 6. The The rationale for bevacizumab use in glioblastoma is that inhibition of VEGF normalizes tumor vasculature, thus decreasing tumor interstitial pressure. This improves access to chemotherapeutic drugs as well as oxygen delivery, thus improving efficacy of radiation therapy \u201312. HoweIt remains unknown if extracranial dissemination is impacted by the interaction of targeted treatments and biological factors such as MGMT methylation status or EGFR amplification . A landmExtrapolation from other disease sites may suggest a potential relationship between MGMT status and ECM \u201319. KohoEGFR gene amplification is another biological factor potentially implicated in ECM of primary brain tumors . EGFR geAs we enter an era of emerging novel therapeutics including immunotherapies and targeted agents, there is potential for unforeseen side effects and unintended detrimental sequelae as a consequence of treatment. While unmethylated MGMT promoter status and amplified EGFR expression, as seen in our case report above, predict a poor prognosis, the relation of these biomarkers to success of targeted antiangiogenic therapies warrants further investigation."} +{"text": "Introduction. Buschke-Ollendorf syndrome (BOS) is an uncommon syndrome characterized by osteopoikilosis and other bone abnormalities, accompanied by skin lesions, most frequently connective tissue nevi. BOS is caused by mutations in the LEMD3 gene, which encodes the inner nuclear membrane protein Man1. We describe a unique case of osteopoikilosis associated with late-onset localized scleroderma and familial LEMD3 mutations. Case Report. A 72-year-old woman presented with adult-onset diffuse morphea and bullous skin lesions. Evaluation revealed multiple hyperostotic lesions (osteopoikilosis) suggestive of BOS. DNA sequencing identified a previously undescribed nonsense mutation (Trp621X) in the LEMD3 gene encoding Man1. Two additional family members were found to have osteopoikilosis and carry the same LEMD3 mutation. Conclusions and Relevance. We report a unique familial LEMD3 mutation in an individual with osteopoikilosis and late-onset morphea. We propose that this constellation represents a novel syndromic variant of BOS. LEMD3 (LEM domain containing 3) gene or in the 1000 Genomes Project database (http://www.1000genomes.org/).Index case DNA was extracted from peripheral blood using a commercial kit . Sanger sequencing of the entire domains scleroderma in childhood ( LEMD3 mutations [ LEMD3 mutation-proven cases of melorheostosis (A review of over 100 published cases of BOS showed that connective tissue nevi (dermatofibrosis lenticularis disseminata) were the most frequent cutaneous manifestation. The diagnosis of BOS was characteristically made before the age of 16. A survey of cases ofhildhood . Howevereostosis have coiLEMD3 mutations show variable penetrance. There is extreme variability in the associated phenotypes, even among individuals harboring identical mutations [ LEMD3 mutation in a specific phenotype is difficult to discern. Although the TGF-\u03b2/Smad signaling pathway plays a pivotal role in both skin and bone homeostasis, it remains unclear how Man1-Smad interactions are affected by the BOS mutations, and whether they contribute to clinical features. While the novel LEMD3 mutation described in this report is predicted to alter the C-terminal domain of Man1 required for R-Smad interactions [\u03b2/Smad signaling in the BOS skin fibroblasts.utations . Given sractions , our funThe coexistence of morphea and lichen sclerosus et atrophicus (LSA) changes is also of note. While this combination has been previously reported as a cause of bullous changes \u201343 in mo LEMD3. We propose that in this case morphea and osteopoikilosis are linked, representing a novel BOS variant that is on the continuum of LEMD3-associated skin and bone manifestations. In light of the known involvement of Man1 in modulating canonical TGF-\u03b2 signaling, we hypothesize that the skin and bone abnormalities associated with LEMD3 mutations might be related to altered TGF-\u03b2 signaling. Future studies will characterize the functional consequences of LEMD3 mutations and their role in the clinical manifestations of the syndrome. Given the diverse phenotypes associated with such mutations and poorly understood mechanisms of how Man1 protein changes contribute to the phenotypic manifestations of BOS, such studies may reveal new roles for this diverse molecule in mesenchymal cell biology.In summary, we describe a case of osteopoikilosis associated with late-onset generalized morphea and associated LSA changes in an elderly individual carrying a previously undescribed familial mutation in"} +{"text": "In addition, the neutralizing effect of Mg(OH)2 was evaluated by degradation study.Biodegradable poly(L-lactic acid) (PLLA) is one of the most widely used polymer in biomedical devices, but it still has limitations such as inherent brittleness and acidic degradation products. In this work, PLLA blends with poly(L-lactide-\u03b5-caprolactone) (PLCL) and Mg(OH)g) of PLLA was slightly reduced from 61 to 52 \u00b0C by adding PLCL additive. Mg(OH)2 in polymeric matrix not only improved the molecular weight reduction and mechanical strength of PLLA, but also neutralized the acidic byproducts generated during polyester degradation.The elongation of PLLA remarkably increased from 3 to 164.4 % and the glass transition temperature One Mg2+ of Mg(OH)2 also needs two anionic compounds to neutralize the charge and thus binds to anionic terminal group of PLLA and/or acidic degradation product; [Mg2+] \u00b7 2[lactic acid\u2013OH\u2212]. The [Mg2+] \u00b7 2[lactic acid\u2013OH\u2212] form inactivates hydroxyl groups of PLLA and byproducts which are capable of backbiting reaction and hydrolysis of ester linkages incorporated into the polymer backbone [2 in PLLA90/PLCL10/Mg5 matrix inhibited consecutive ester pyrolysis and thermal-hydrolysis reactions with binding anionic terminal group of polymer and acidic byproducts generated during thermal processing. However, the conclusive mechanism was not fully understood yet. The inorganic Mg(OH)2 particles slightly enhanced the tensile strength affected by molecular weight, but reduced the elongation at break due to poor interfacial adhesion, as shown in Fig.\u00a02 and PLCL, respectively, compared with PLLA control. Mg(OH)2 not only affected the molecular weight reduction and mechanical strength under thermal processing, but also alleviated reduced pH value and molecular weight during degradation.The PLCL (75:25) and Mg(OH)backbone \u201334. It sg value below 60 \u00b0C. Meanwhile, the Mg(OH)2-containing matrix such as PLLA100/Mg5 and PLLA90/PLCL10/Mg5 slightly compensated the pH as compared with PLLA control, corresponding to 7.1 in 14 days. For pH neutralization of the degradation medium, many researchers studied some inorganic compounds such as sodium bicarbonate and calcium carbonate and incorporated these inorganic compounds into the PLLA matrix to evaluate their effect on the degradation process [2 particles can neutralize the acidic environment since the dehydrated magnesium ion can bind with two moles of anionic compounds [2 particles, and also the accelerated formation of degradation byproducts was prevented by binding of acidic terminal groups of PLLA and PLCL to Mg(OH)2.Figure process . In partompounds . The aci2 on molecular weight reduction as well as pH balance 2 hindered the accumulation of acidic byproducts and averted backbiting and intermolecular transesterification. These results demonstrated that the Mg(OH)2 affected the prevention of molecular weight reduction by neutralizing acidic substance.The molecular weights of degraded samples were evaluated to confirm the effects of Mg(OH)nce Fig.\u00a0. The PLLg medium . The aci2 additives successfully fabricated by thermal processing, and the effects of PLCL and Mg(OH)2 additives on physico-chemical and thermal properties under thermal decomposition of PLLA matrix were assessed using various analyses. With increasing the amount of PLCL, the PLLA/PLCL blends exhibited the alleviation of molecular weight reduction and the improvement of flexibility compared with thermally processed PLLA control. The PLLA/PLCL blends with increasing proportion of polycaprolactone depicted proper thermal stability as well as mechanical strength including tensile strength and elongation. In Mg(OH)2-containing matrix, the molecular reduction and mechanical strength were dramatically improved because the dehydrated magnesium ion inhibited the decomposition of polyester substrate by counteracting acidic compounds, which is a kind of nucleophile at ester linkage. In particular, the Mg(OH)2 in matrix certainly neutralized the acidic byproducts involved during polyester degradation, that caused the acid-induced inflammatory reaction in vivo. The obtained results suggested that PLCL and Mg(OH)2 additives were effective to enhance flexibility and control degradation behavior of biodegradable PLLA matrix, and therefore the PLLA/PLCL/Mg(OH)2 composites have the potential as a material for bio-absorbable biomedical devices such as implants and stents.The new PLLA blends with PLCL and Mg(OH)"} +{"text": "Firefighting is a hazardous task associated with a heavy workload where task duration may be limited by air cylinder capacity. Increased fitness may lead to better air ventilation efficiency and task duration at a given heavy work intensity.2peak: 47.8 \u00b1 5.1 mLO2.min-1.kg-1) completed the following tests on 3 different days while wearing firefighting protective clothing (FPC), self-contained breathing apparatus (SCBA) and air cylinder: 1- The graded walking test (GWT) for measuring different physiological parameters while connected to a metabolic system (gas exchanges); 2- The 10 METS treadmill test (T10) designed to measure the time to ventilate air from the cylinder at 10 METS, the intensity needed to complete the fire fit test work simulation described by Deakin et al. (1) within 8 min (2); 3- The simulated work circuit (SWC) to measure the time needed to perform a test mimicking different firefighting tasks while wearing FPC and breathing with the SCBA. Participants performed the SWC as quickly as possible while respecting regulations of the test protocol. Moreover, skeletal muscle oxygenation was measured during all three tests.Thirteen male firefighters , better peak oxygen consumption during the GWT and performed longer until exhaustion on the GWT . Participants who completed the SWC more rapidly and reached a higher VO2peak also had lower VE and VE/VO2 values during submaximal workload on the GWT. Moreover, they had greater skeletal muscle deoxygenation during the SWC .Firefighters who performed the SWC in a shorter time had lower air cylinder ventilation values on the T10 .Greater aerobic fitness was associated with greater air ventilation efficiency of faster firefighters on the SWC. According to Holm\u00e9r and Gavhed (3), cardiovascular strain is lower in individuals with higher maximal aerobic capacity for a given submaximal intensity. Moreover, correlation between SWC completion time and HHb suggests that better aerobic fitness enhances deoxygenation in the These results demonstrate that the fastest participants on the SWC had better air ventilation efficiency that could prolong interventions in difficult situations requiring air cylinder use. Moreover, the fastest participants had a greater skeletal muscle deoxygenation during the SWC."} +{"text": "Elective intracranial pressure (ICP) monitoring is a useful tool in the diagnosis and evaluation of simple and complex cerebrospinal fluid dynamic disturbances. Whilst many previous research papers have focused on patients undergoing ICP monitoring acutely following traumatic brain injury (TBI), few have looked into the duration of monitoring required to achieve an accurate picture of a patients intracranial dynamics in non acute, elective cases. At our institution we currently complete monitoring for a period of >48hrs.A retrospective audit, assessing any patient admitted electively to our institution for ICP monitoring over a 3 month period. Exclusion criteria included acute admissions and patients who underwent a change in their treatment whilst undergoing ICP monitoring . ICP results were analysed focusing on median ICP and Median pulse amplitude over three time periods: total data collected v first 48hrs of data collection v first 24hrs of data collection.18 patients met the desired criteria. Mean length of monitoring was 3 days (range 2-5) for the total number of patients. There was no significant difference between 24hrs and 48hrs duration of monitoring for the median ICP (p=>0.05) and ICP pulse amplitude (p=>0.05).24 hour monitoring of ICP in elective patients in a stable condition without changes to their current treatment is sufficient to detect mean ICP and pulse amplitude. Further studies may be appropriate to assess if fewer than 24hrs monitoring can also prove an accurate method of monitoring ICP."} +{"text": "Plasmodium falciparum (Pf) incorporate active infection detection strategies to target the subclinical transmission reservoir. Currently available immunochromatographic lateral flow tests lack the sensitivity required for Pf malaria elimination; the limit of detection (LOD) of these rapid diagnostic tests (RDTs) is above that which is required to identify all transmissible infections. Malaria RDTs with a significantly improved LOD would enable more effective elimination interventions while retaining the critical advantages of low cost, ease of use, and rural deployment capability. Histidinerich protein 2 (HRP2) is a high-priority target analyte for identifying individuals at risk of transmitting Pf. HRP2 protein is secreted by Pf parasites in relatively high concentrations, is widely conserved among Pf strains, and persists in human plasma for up to four weeks. These characteristics allow HRP2 antigen detection even in the absence of circulating parasites during cyclical sequestration. However, the highly polymorphic nature of HRP2 makes optimization of RDTs challenging. Current HRP2-detecting RDT reagents primarily target Type 2 (AHHAHHAAD) and Type 7 (AHHAAD) HRP2 motifs, and independent antibody discovery confirms the most common selection of a C1-13/PTL-3 antibody conjugate pair yields the best RDT performance for the majority of parasite strains. In scenarios where malaria transmission rapidly decreases, exclusive use of a diagnostic whose sensitivity varies with the number of HRP2 epitope repeats can result in single axis evolutionary pressure, increasing selection for less-detectable parasite strains. To evaluate the feasibility of developing an HRP2-based RDT that is highly effective in mass testing and treatment programs, we have assessed the technical characteristics of HRP2 reagents. Our aim is to facilitate the development of improved malaria HRP2 RDTs by: (1) incorporating an understanding of HRP2 structure-function relationships and their impact on epitope-antibody interactions and adaptation, (2) correlating HRP2 concentration levels (in pg/mL) with traditional malaria diagnostic metrics (parasites/mL) for comparative evaluation of tests, and (3) predicting optimal capture reagents to improve and sustain HRP2 RDT performance. Based on our ongoing analysis, we present a product development strategy that combines capture reagent and platform innovations that can be used to make highly sensitive Pf malaria RDTs. In addition, we present criteria for the rational selection of HRP2 protein and parasite culture controls to support the product lifecycle from development through commercialization and scale-up.Malaria programs aimed at eliminating"} +{"text": "Tenofovir is a commonly used component of antiretroviral therapy (ART) to reduce vertical transmission of HIV. Although systematic review of tenofovir use in pregnancy concluded it to be low risk for foetal abnormalities , data isWe retrospectively analyzed data on renal function in pregnancy from a cohort of women attending a busy inner city London antenatal clinic. All women were screened for renal function throughout pregnancy via serum creatinine and estimated glomerular filtration rate (eGFR) calculated using modification of diet in renal disease (MDRD) and corrected for ethnicity.Ninety-seven HIV-1 positive women were registered at Homerton Hospital antenatal service of a total of 105 pregnancies between January 2010 and September 2013. Tenofovir was prescribed in 71/105 pregnancies (67.6%). Of the 71 pregnancies, 41 were prescribed tenofovir pre-conception (57.7%). Of the pregnant women who started tenofovir in pregnancy, 21/31 (67.7%) were initiated before week 24 of pregnancy, in line with British HIV association (BHIVA) guidelines [Consistent with current guidelines and experience, this study shows tenofovir did not cause decline in renal function in pregnancy in our cohort of HIV-1 positive women, whether started during pre-conception or during pregnancy. More evidence should be prospectively collected looking at effects of tenofovir on other measures of tubular renal function in pregnancy such as proteinuria and protein-creatinine ratio."} +{"text": "Sulfite hypersensitivity (SH) may have different clinical presentations, ranging from mild cutaneous symptoms to anaphylaxis. Our aim was to evaluate the prevalence and clinical characteristics of individuals with suspected sulfite allergy in our Food Allergy Unit (FAU).\u00ae capsules) was performed.Clinical files of the 335 patients referred to the FAU of a Portuguese University Hospital for suspected food allergic reactions between January 2010 and April 2014 were retrospectively reviewed; those with suspicion of SH were selected. Patients\u2019 demographics, medical history and SH diagnostic procedures reviewed. The allergy work-up was considered complete when an open food challenge (OFC) with 390mg of sodium metabisulfite of the patients were referred due to suspected SH. They had a median (inter quartile range) age of 32 years, 52% were female, 42% atopic, 29% had asthma, 29% rhinitis and 16% chronic urticaria. Involved foods were: wine (19 reactions); canned food (11); seafood (8); bottled soft drinks (7) and processed food (e.g.: pizza (5), delicatessen meats (3)). The most common manifestations were urticaria (84%) and dyspnea (36%); 3 individuals reported anaphylaxis. Patients with asthma had a higher proportion of respiratory symptoms (p=0.020). All except 4 individuals completed the study with OFC. Twenty-five (93%) had negative OFC (1 had a mild adverse event (emesis) not consistent with the previous reactions). Two (7%) women with a history of urticaria and angioedema (one with concomitant dyspnea) after wine and bottled beverage ingestion had positive OFC (both at a cumulative dose of 190mg); they presented facial and oral pruritus and urticaria; the one with previous respiratory complaints also presented coughing and emesis.In our sample, the prevalence of sulfite hypersensitivity suspicion was low and sulfite allergy was ruled out in the majority; only 2 patients had positive oral challenges. When sulfite hypersensitivity is considered, oral food challenge with sulfite is both a safe and valuable tool in the diagnostic work-up."} +{"text": "The bovine leukemia virus (BLV), a retrovirus structurally and functionally related to the human T-lymphotropic viruses HTLV-1 and HTLV-2, is the etiological agent of bovine leucosis. B cell immune mechanisms may play a major role in protection against BLV infection. A battery of 157 synthetic peptides, 15-mer length, 4 amino acids overlapped, was used to mapping B cell epitopes on BLV envelope glycoprotein gp5l and capsid protein by peptide microarrays. Two susceptible cattle with the homozygote BoLA-DRB3 *1601 allele and two non-susceptible cattle with DRB3 *1501/*2703 alleles and DRB3 *1501/*0503 alleles were infected with BLV, and serums were collected from those cattle each of having challenged BLV before and after. Only one epitope A out of 157 synthetic peptides responds with all of four BLV positive serums not their negative serums. To demonstrate whether epitope A is common B cell epitope or not by our established peptide ELISA system that uses maleimide activated mariculture keyhole limpet hemocyanin (mcKLH) carrier protein. To found, there are 7 kinds of BLV positive serum no response with epitope A among 232 kinds of positive serum. Furthermore, we searched other peptides respond with BLV positive serum that contain 7 kinds of serum no responded with epitope A and found epitope B among peptides of non-specifically responded on peptide microarray. Epitope B strongly responded with all of 232 kinds of serum as estimated by peptide ELISA system. Our results shows that epitope B has a tendency to react some BLV negative serums, thus we will perform further experiments to determine the common B cell epitope just responses with positive serum by peptides that are 3 alanine substitutions as shifting one amino acid on epitope sequence."} +{"text": "Plasma neutrophil geletinase-associated lipocalin (NGAL) has been regarded as one of the valuable markers of development of acute kidney injury (AKI) in ICU admitted patients. However, clinical significance of short-term change of plasma NGAL was not clearly investigated.50 patients who admitted intensive care unit (ICU) was included by prospective manner. We analyzed plasma NGAL level by Triage immunoassay on ICU admitted time and 12 hours after. Change of estimated glomerular filtration rate (by MDRD equation) and need of renal replacement therapy was investigated. Patients with chronic kidney disease were excluded.0-12 hr was independent predictors of unfavorable renal outcome. However, \u0394NGAL0-12hr did not significantly correlated with the change of glomerular filtration rate between day 1 and day3.Persistent elevation of plasma NGAL above 1000 ng/mL on 0 and 12 hours was significantly associated with need for early renal replacement therapy. Patient with increased level of NGAL at 12 hours showed unfavorable outcome which is related to renal replacement therapy or progressive renal impairment . Conversely, 2/17 (11.8%) of patients had poor renal outcome when NGAL decrease more than 20% from baseline at 12 hours after ICU admission. On multivariate analysis, baseline NGAL and \u0394NGALOur data suggest that measurement of early serum NGAL change may be useful marker in predicting the renal impairment and need for renal replacement therapy in intensive care unit."} +{"text": "Bile acids (BA) are signaling molecules with pleiotropic paracrine and endocrine functions is a ligand activated transcription factor responsive to different bile acids and highly expressed in hepatocytes. FXR plays an important role in the regulation of bile acid synthesis, detoxification and secretion. Activation of FXR contributes to liver regeneration following partial hepatectomy and alleH4 is a lTGR5 Gpbar-1, M-Bar) is a GPCR responsive to various unconjugated and conjugated bile acids with taurine-conjugated secondary bile acids, such as taurolithocholic acid (TLC) and taurodeoxycholic acid (TDC), being the most potent TGR5 ligands , M-Bar i. ExpressAim of our studies is to elucidate the role of TGR5 in bile acid induced liver damage and regeneration. TGR5 knockout and wildtype mice were fed ad libitum with a bile acid containing diet for 7 days. Liver injury was assessed by serum biochemistry and liver histology. Cellular proliferation was made visible using immunohistochemistry of liver sections and an antibody against proliferating cell nuclear antigen (PCNA). Ductular proliferation was assessed and quantified by cytokeratin (CK)-19 immunofluorescence staining and confocal laser scanning microscopy. Cholangiocytes were cultivated from isolated ducts from livers of TGR5 wildtype and knockout mice and cell proliferation was measured by BrdU incorporation after stimulation with bile acids or specific TGR5 agonists. The role of TGR5 downstream signalling pathways was analyzed with different kinase inhibitors. The generation of reactive oxygen species (ROS) was measured using a fluorescent dye and shedding of EGF was determined by an ELISA assay. Western blotting was carried out to confirm the phosphorylation of the EGFR and ERK1/2. Bile acid feeding resulted in more elevated liver function tests in TGR5 knockout mice as compared to their wildtype littermates on the same diet. Livers from TGR5 knockout mice fed with bile acids for 4 days showed larger areas of hepatocytes necrosis as livers from wildtype animals on the same diet. In contrast, livers from bile acid-fed wildtype animals displayed increased PCNA-positive hepatocytes and cholangiocytes, as compared to the livers from the respective knockout animals, indicating a rise in cell proliferation following bile acid feeding in wildtype livers. Since TGR5 is highly expressed in cholangiocytes, we focused on bile acid mediated cholangiocyte proliferation. While the amount of CK-19 positive bile ducts was comparable between TGR5 wildtype and knockout mice on chow diet, a significant increase of bile duct proliferation as measured by CK-19 staining was detected in livers from wildtype mice following 7 days on CA diet as compared to livers from TGR5 knockout mice on the same diet.Incubation of isolated cultivated cholangiocytes from TGR5 wildtype and knockout mice with taurolithocholic acid (TLC 10 and 25 \u00b5M) or TGR5 specific agonists led to a significantly increased cholangiocyte proliferation exclusively in wildtype derived cells. Preincubation of wildtype derived cholangiocytes with inhibitors of Src kinases, epidermal growth factor receptor (EGFR) kinase or ERK1/2 kinases significantly reduced TLC and TGR5 agonist mediated cell proliferation as measured by BrDU incorporation. Treatment with inhibitors of ROS formation, such as N-acetylcystein or apocynin also suppressed TLC dependent BrdU incorporation. In contrast, inhibition of adenylate cyclase by SQ22536 or dideoxyadenosine showed no effect on TLC or TGR5 agonist mediated cholangiocyte proliferation. Stimulation of wildtype derived cholangiocytes with TLC and a TGR5 agonist significantly elevated ROS formation. Increased shedding of EGF was also detected after incubation of wildtype cholangiocytes with TLC or the TGR5 agonist. Furthermore, TGR5 activation significantly induced tyrosine phosphorylation of the EGFR at amino acid positions 845 and 1045 and increased ERK1 and ERK2 phosphorylation. Thus, bile acids mediate cholangiocyte proliferation through a TGR5-ROS-Src-EGFR-ERK signaling pathway, which is independent of adenylate cyclase activation.In summary, TGR5 knockout mice show reduced hepatocyte and cholangiocyte proliferation and more pronounced liver injury in response to bile acid feeding. This is in line with a recent publication, demonstrating that liver regeneration is impaired in TGR5 knockout mice following partial hepatectomy ."} +{"text": "The combination of a global demographic shift and increased survival following critical illness has led to an increasing number of patients requiring prolonged mechanical ventilation (PMV) and longer critical care stay. This is a prospective observational study evaluating the characteristics and speciality-based outcome of critically ill patients undergoing prolonged mechanical ventilation in the North of England Critical Care Network (NoECCN).A weekly survey was conducted over a 1-year period screening patients older than 16 years of age requiring PMV in all 18 adult critical care units within the NoECCN. Patient data collected included patient demographics, admission diagnosis and speciality, hospital length of stay (LOS) pre and post critical care admission, severity of illness scores, critical care LOS and status at hospital discharge.During the study period 134 patients met the criteria for PMV representing 1% of annual admissions and 6.9% NoECCN bed-days. The majority of patients receiving PMV were medical (50.7%), followed by emergency surgery (20.1%), elective surgery (16.4%) and specialist services such as spinal cord injury (8.2%) and cardiothoracic transplant (4.5%). The commonest admission diagnosis in the medical population was pulmonary infection followed by acute neurological disorders, while 89.4% of surgical patients were admitted to critical care during the perioperative period. At the end of the study period the highest hospital mortality was observed in the nonspecialist surgical population (26.5%). In contrast, the medical population had one of the lowest hospital mortality rates (11.8%), lower than predicted using the intensive care national audit research network illness severity score. Comparable rates of hospital discharge were found in both medical (85%) and nonspecialist surgical patients (88.9%).The results of this study highlight an expanding proportion of NoECCN critical care bed-days occupied by stable patients undergoing PMV. In keeping with published UK data, elevated hospital mortality was observed in the nonspecialist surgical subpopulation. Although the literature suggests the medical cohort of patients has poorer prognosis, within our region all were liberated from mechanical ventilation and over 80% were discharged from hospital."} +{"text": "Haemophagocytic lymphohistiocytosis (HLH) is a severe condition in which there is extreme uncontrolled inflammation, and may progress rapidly to multi-organ failure and death. HLH may be genetic (primary HLH), or secondary to infection or autoimmune/ autoinflammatory conditions; if the latter, it is also referred to as macrophage activation syndrome (MAS). Distinguishing between primary HLH and MAS is challenging but important since the former requires different therapeutic approaches including allogeneic haematopoietic stem cell transplantation (HSCT) for long-term survival. HLH screening tests are now being used in patients presenting with suspected MAS. In systemic Juvenile Idiopathic Arthritis (sJIA), some patients demonstrate temporary perforin expression abnormalities that resolve with disease control. The utility of other screening tests in a rheumatology context is unknown.The purpose of this study was to describe the performance of screening tests used in the HLH/MAS work up of children presenting to a specialist paediatric rheumatology centre, and review outcomes of those with screening abnormalities.A database exists of patients who had screening tests for suspected HLH/MAS. Screening tests (flow cytometry) included: intracellular expression of perforin in CD56+ Natural Killer (NK) cells; CD107a Granule Release Assay (GRA) in response to PHA in NK cells or anti-CD3 stimulation of CD8 lymphocytes; in male patients Signal Lymphocyte Activating Molecule Associated Protein and X-linked Inhibitor of Apoptosis Protein expression. All tests requested by paediatric rheumatology over a 5 year period (2007-2011) were included. Patient records and laboratory parameters were retrospectively reviewed.22 patients , median age 6.5 years (range 0.6-16) underwent screening tests, with median follow-up of 16 months (range 3-51). At presentation only 2/22 (9%) clinically met HLH criteria. Screening results were available for 20 patients; 7 (35%) had at least one persistent abnormality in any one of the tests; this group was associated with 57% mortality or need for HSCT, compared to 8% with no abnormality on any of the tests (p = 0.03). 6/20 (30%) had persistently abnormal GRA: their final diagnoses were sJIA with MAS (n = 3); primary HLH (n = 2); and overlap syndrome (n = 1). 1/4 boys screened for XIAP had absent expression with subsequent genetic confirmation of XLP2. 18 patients had perforin screened, and 5 boys were screened for SAP expression; all had normal results.Primary HLH and MAS may overlap clinically and screening in patients with suspected MAS is warranted; overall 14% had an eventual diagnosis of primary HLH. Persistently abnormal GRA defines a high risk group with poor outcome possibly due to an unidentified HLH gene. The effect of immunosuppression on the GRA was not assessed. Further research is required in those with abnormal GRA to help understand the pathogenesis of HLH/MAS.None declared."} +{"text": "Advance directives (AD) and/or a health care surrogate decision maker (HCS) are potentially helpful for caregivers to respect the patients' autonomy whenever their competence is affected. Patients planned for major cardiovascular surgical procedure requiring intensive care may consider AD/HCS as important or necessary because of the coming exposure to a potential life-threatening situation.To investigate whether a major cardiovascular surgical procedure requiring intensive care impacts on patients', interests for AD/HCS.gA) met the day before and after ICU discharge, or to a group B (gB) met only after ICU discharge. At each meeting, they were interviewed according to the same questionnaire.Patients planned for major cardiovascular surgery were randomized either to a group A (361(89%) patients (of 405 eligible) were interviewed. Male: 256(71%); age(mean \u00b1 SD):68 \u00b1 15 years. 95(27%) had a last will, 77(21%) a life insurance, 119(33%) a funeral plan and 43(12%)an organ donor card. 181(50%) patients were randomized in the gA, 180(50%) in the gB.After surgery, 164(91%) of the gA patients remembered the interview - before, 90(50%) what AD are and 61(34%) could give a correct definition of AD.Few patients, even when scheduled for major surgical procedure, knew what AD or HCS are and even fewer had AD/HCS. Their incidence was much lower than other plans for the future . Undergoing major surgery requiring intensive care modified significantly the attitudes of patients towards AD/HCS, decreasing their interest. Further analyses regarding these patients' reasons for or against AD/HCS will provide more information to understand the rarity of advance care planning.This study is sustained by the FNRS (CR31I3_127135/1)"} +{"text": "Athletes use ergogenic aids in an attempt to increase training-adaptations, which serves to enhance their performance during competition. Creatine monohydrate is one of the most studied ergogenic aids. Although many studies have reported the efficacy and effectiveness of creatine monohydrate supplement manufacturers continually introduce newer forms of creatine into the marketplace. The newer forms of creatine purport to be more effective than creatine monohydrate alone. However, there is little evidence to support most manufacturers' claims.We examined 28d of randomly assigned (1) placebo (PL), (2) Creatine monohydrate , (3) creatine nitrate and (4) CrN2X on bench press performance. Participants presented for fasting (12 h) testing after abstaining from exercise and alcohol for 48 h. Performance (reps at 70% of bench press 1 RM) was measured using a Tendo Fitrodyne at 0 & 28d and analyzed by MANOVA or one-way ANOVA. Mean changes (95% CI) were reported.FASEB J, 29(1):LB248, 2015) that all treatment groups increased bench press repetitions after 28d of supplementation; however, total work (reps \u00d7 weight lifted) during bench press was greater at 28d for CrN2X vs. CrN and PL . MANOVA univariate analysis of power data indicated a significant time effect with all power output variables , average power (AP), and average velocity (AV)). No significant group by time effects were observed among groups. One-way ANOVA of the 3rd set of exercise performed to exhaustion revealed no significant differences among groups in changes from baseline after 28d of supplementation. However, pairwise comparison of 95% CIs revealed a significant difference in peak power and average power between CrN2X and PL and CrN2X and PL , respectively. Average power was also significantly different between CrN2X and CrN . Average velocity during bench press test was also significantly different between CrN and PL .We previously reported (Results suggest some ergogenic value of consuming these types of creatine containing pre-workout supplements on bench press power adaptations during training in comparison to PL responses."} +{"text": "Ruffs (Philomachus pugnax) exhibit major dark/light color differences in melanin-based male breeding plumage which is closely associated with alternative reproductive behavior. A previous study identified a microsatellite marker (Ppu020) near the MC1R locus associated with the presence/absence of ornamental plumage. We investigated whether coding sequence variation in the MC1R gene explains major dark/light plumage color variation and/or the presence/absence of ornamental plumage in ruffs. Among 821bp of the MC1R coding region from 44 male ruffs we found 3 single nucleotide polymorphisms, representing 1 nonsynonymous and 2 synonymous amino acid substitutions. None were associated with major dark/light color differences or the presence/absence of ornamental plumage. At all amino acid sites known to be functionally important in other avian species with dark/light plumage color variation, ruffs were either monomorphic or the shared polymorphism did not coincide with color morph. Neither ornamental plumage color differences nor the presence/absence of ornamental plumage in ruffs are likely to be caused entirely by amino acid variation within the coding regions of the MC1R locus. Regulatory elements and structural variation at other loci may be involved in melanin expression and contribute to the extreme plumage polymorphism observed in this species.Sequence variation in the Philomachus pugnax) is a lekking sandpiper which exhibits major dark/light color differences in melanin-based male breeding plumage that is closely associated with a genetic polymorphism for alternative male mating behavior gene explains major dark/light plumage color variation in several avian species as well as coat color variation in many mammals (MC1R gene perfectly associate with dark and light plumage types in bananaquit (Coereba flaveola), lesser snow goose (Anser c. caerulescens), Arctic skua (Stercorarius parasiticus) and chestnut-bellied monarch (Monarcha castaneiventris) (Faeder locus) . In ruffr locus) was assos) . Sequences were aligned in MEGA.v5.0 (Genomic DNA was extracted from blood samples stored in absolute ethanol (50 \u03bcL of blood in 1.5mL of absolute ethanol) using an ammonium acetate precipitation method . A segmed MSHR74 . Each 10d MSHR74 . Purified MSHR74 and Phrad MSHR74 softwareEGA.v5.0 using ClMC1R gene corresponding to positions 79\u2013899 of the aligned chicken MC1R gene sequence and 1 nonsynonymous substitution (A/G: His207Arg); the latter polymorphism is shared with dark/light plumage differences in the red-footed booby (Sula sula) . At all omorphic . None ofomorphic .MC1R does not explain the major plumage color differences of male ruffs, we conclude that the control of plumage variation in ruffs is more complex than in other bird species with simple dark and light morphs. It is unlikely that the presence of dark or light coloration or the presence/absence of ornamental plumage in male ruffs is solely determined by amino acid variation within the MC1R locus. Because only a partial amino acid sequence of the coding region of the MC1R gene was sequenced, we cannot rule out the possibility that functional non-synonymous substitutions affecting plumage color may be present in the regions (approximately 9% based on other bird species) not sequenced. However, it seems more likely that regulatory polymorphisms account for the observed variation. The extensive individual variation suggests that ornamental plumage coloration is a polygenic trait, which might nonetheless involve MC1R along with other genes that affect the deposition of melanin ; UK Biotechnology and Biological Sciences Research Council grant (BB/J018937/1 to T.B.); NSERC PGS-D3 (L.L.F.); and Marie Curie Intra-European Fellowship (C.K.)."} +{"text": "We used a target-centric strategy to identify transporter proteins upregulated in pancreatic ductal adenocarcinoma (PDAC) as potential targets for a functional imaging probe to complement existing anatomical imaging approaches. We performed transcriptomic profiling (microarray and RNASeq) on histologically confirmed primary PDAC tumors and normal pancreas tissue from 33 patients, including five patients whose tumors were not visible on computed tomography. Target expression was confirmed with immunohistochemistry on tissue microarrays from 94PDAC patients. The best imaging target identified was SLC6A14 . SLC6A14 was overexpressed at the transcriptional level in all patients and expressed at the protein level in 95% of PDAC tumors. Very little is known about the role of SLC6A14 in PDAC and our results demonstrate that this target merits further investigation as a candidate transporter for functional imaging of PDAC. Early detection and surgical resection of pancreatic ductal adenocarcinoma (PDAC) confined to the pancreas offers the best hope for cure or extension of lifespan. Recent breakthroughs in serum profiling, most notably mass spectral and antibody array technologies, provide hope for screening patients with asymptomatic disease , 2. Howe18F]fluoro-D-glucose positron emission tomography (18FDG-PET), combined with CT or MRI, is a highly sensitive diagnostic tool for many tumor types, but its utility in PDAC is hampered by low tumor signal-to-background ratios that limit its sensitivity for detection of lesions below the realized resolution of PET (approximately 1\u2009cm). A new functional imaging probe that selectively targets PDAC with high sensitivity is a critical unmet need in PET/CT or PET/MRI that could transform patient management by allowing earlier PDAC detection and surgical intervention and that could improve preoperative staging of disease to decrease the number of unwarranted surgeries in patients who might benefit from experimental systemic therapy.The majority of large PDACs are detected with anatomical imaging techniques such as computed tomography (CT), magnetic resonance imaging (MRI), and ultrasound. Multidetector, helical CT with intravenous administration of contrast material is the most commonly used imaging procedure to detect and stage suspected PDAC. Diagnostic accuracy decreases, however, with decreasing tumor size \u20135 and in131I (iodine 131) radiotherapy for thyroid cancer. Another well-characterized SLC family member useful in functional imaging is SLC2A1 (GLUT1), a major glucose membrane transporter that is upregulated in tumor cells dihydroxyphenylalanine [11C-methyl]methionine fluoroethyl)methylamino)ethyltyrosine or ([18F]FEMAET), a cationic amino acid PET probe that demonstrates positive uptake in SLC6A14-positive xenografts [Further understanding of the function, substrate specificities, and tissue distribution of SLC6A14 will aid in the potential future development of a selective functional imaging probe or the discovery of targeted therapeutic agents. An exciting development towards this goal was the recent synthesis ofnografts , 41. Ant O-2((2-Ffluoroetnografts .SLC6A14 appears to be a good candidate transporter for further exploration as a functional imaging target for PDAC that could complement existing anatomical imaging techniques for both diagnosis and staging."} +{"text": "Women diagnosed with breast cancer are at increased risk of contralateral breast cancer; some of these cancers will be synchronous and mammographically occult (M1). Ultrasound may detect M1 breast cancers but also benign lesions that necessitate needle testing, conferring additional patient morbidity that could be termed 'over investigation'. Local guidelines for ultrasound of the M1 contralateral breast vary between units. We present a retrospective audit of contralateral M1 breast ultrasound within our screening assessment clinics.Screening and pathology hospital databases of 2013 and 2014 identified records of 331 women with screen-detected breast cancer. Descriptive statistics were performed.All 331 women underwent ipsilateral breast ultrasound; 288 (87 %) underwent ultrasound of their contralateral mammographically normal (M1) breast. Six contralateral breast lesions were needle sampled: four B2 lesions, two B3 without atypia. No subsequent breast cancer has been detected in any of these patients to date.Two years of routine contralateral ultrasound has yielded no cancers but also very few benign biopsies. Ongoing audit and discussion of risk/benefit to patients is indicated."} +{"text": "Data on serum lipids and lipoproteins were collected at DCCT baseline (1983\u201389) and were correlated with common and internal carotid IMT determined by ultrasonography during the observational follow-up of the DCCT, the Epidemiology of Diabetes Interventions and Complications (EDIC) study, at EDIC \u2018Year 1\u2019 (199\u20131996) and EDIC \u2018Year 6\u2019 (1998\u20132000). This article contains data on the associations of DCCT baseline lipoprotein profiles with carotid IMT at EDIC Years 1 and 6, stratified by gender. The data are supplemental to our original research article describing detailed associations of DCCT baseline lipids and lipoprotein profiles with EDIC Year 12 carotid IMT Type 1 diabetes (T1DM) is associated with increased risk of macrovascular complications. We examined longitudinal associations of serum conventional lipids and nuclear magnetic resonance (NMR)-determined lipoprotein subclasses with carotid intima-media thickness (IMT) in adults with T1DM ( Specifications TableValue of the data\u2022Previously unreported longitudinal associations of baseline lipoprotein subclasses with carotid IMT at two follow-up time-points in a large well-characterized cohort of people with T1DM.\u2022May stimulate further research on the clinical utility of lipoprotein subclasses as prognostic markers for cardiovascular disease in diabetes.\u2022May facilitate new therapies to target lipids and lipoprotein classes significantly associated with vascular events in diabetes.1The Diabetes Control and Complications Trial (DCCT) examined the effects of intensive diabetes management on the development and progression of diabetic retinopathy The baseline characteristics of the participants have been described previously https://www.niddkrepository.org/home/). This website includes information regarding the data sets that are available, and details of the data request, review, and approval process.22.1n=730) or intensive (n=711) diabetes treatment The original DCCT cohort comprised 1441 T1DM participants aged 13\u201339 years at study entry (1983\u20131989). They had no dyslipidemia or hypertension and were randomly assigned to conventional using a 400-MHz proton NMR analyzer at LipoScience Inc. as described 2.41c was measured by high-performance ion exchange liquid chromatography Total cholesterol, triglyceride, and HDL-C levels were determined using previously reported enzymatic methods 2.51c, statin use and ultrasound imaging device. Two-tailed p<0.05 was considered to be statistically significant. Data were analyzed using SAS/STAT software .Multiple regression analyses were performed to examine correlations of conventional lipids and NMR-LSP at DCCT baseline with common and internal carotid IMT at EDIC Years 1 and 6, stratified by gender. Each lipid/lipoprotein measure was included as an independent variable in the linear model simultaneous with a fixed group of covariates that were measured at DCCT baseline: diabetes duration, smoking (yes/no), DCCT treatment group, body mass index (BMI), urinary albumin excretion rate (AER), and HbA"} +{"text": "Telaprevir (TVR) plus peg interferon (PEG-IFN) and ribavirin (RBV) substantially increase treatment efficacy for genotype 1 chronic hepatitis C virus (HCV) infection but data about its safety in HIV patients with cirrhosis are lacking. Our purpose was to evaluate estimated Glomerular Filtration Rates (eGFR) variations during combination therapy in a difficult-to-treat HIV/HCV population with advanced fibrosis/cirrhosis through three different scores commonly used in clinical practice: CKD-EPI (Chronic Kidney Disease Epidemiology Collaboration), Modification of Diet in Renal Disease (MDRD) scores and MDRD 6 variable . Second objective was to identify any association between creatinin clearance and haemoglobin (Hb) variation during combination therapy.We conducted an observational retrospective study including 18 HIV/HCV patients attending our clinic who started combination therapy for HCV, including in the analysis the first 16 weeks of therapy. Treatment included TVR 1125 mg BID (twice a day) for 12 weeks, PEG-IFN \u03b1-2a 180 mcg QW and RBV daily dose according to body weight . Advanced fibrosis was defined as Metavir score=F3 or Ishak 3\u20134 and cirrhosis was defined as Metavir score F4 or Ishak 5\u20136 assessed by liver biopsy or transient elastography respectively. P per trend were assessed to evaluate any change of SCL and eGFR (according to different formulas). Multilevel linear regression analysis was performed to estimate factors associated with reduction of Hb.Baseline characteristics and HCV virological responses were collected. In our experience, combination therapy with TPV in cirrhotic HIV-HCV patients did not significantly affect renal function. As expected, Hb levels decreased during treatment and it is likely related to RBV exposure. In multivariable analysis, reduction of Hb appeared to be related to SCL higher levels, thus suggesting even a mild decrease of renal function."} +{"text": "Coronary artery disease (CAD) and left ventricular hypertrophy are prevalent in the chronic kidney disease (CKD) and renal transplant population. Advances in cardiovascular magnetic resonance (CMR) with the blood oxygen level-dependent (BOLD) technique provides unprecedented capability to assess myocardial oxygenation as a measure of ischaemia. We hypothesised that myocardial oxygenation would be reduced in advanced CKD and renal transplant patients and may provide a novel strategy for assessing myocardial ischaemia.We prospectively studied 20 advanced CKD subjects ml/min and 12 CKD group with median eGFR 14 (range 8-18) ml/min), 8 renal transplant (RT) recipients with median eGFR 74.5 (range 57-114) ml/min and 7 hypertensive (HT) controls with median eGFR 107 (range 57-144) ml/min. All patients were asymptomatic for CAD and none had prior history of CAD. All groups had cine and BOLD CMR at 3T, and RT and HT groups also had late gadolinium CMR to assess infarction/replacement fibrosis. CKD group additionally underwent 2D echocardiography strain to assess fibrosis. Myocardial oxygenation was measured at rest and under stress with adenosine (140 \u00b5g/kg/min) using BOLD Signal Intensity (SI). Analyses were performed using linear mixed models.A total of 1074 myocardial segments of the advanced CKD group [522 myocardial segments of dialysis group and 552 myocardial segments of CKD group], 456 myocardial segments of RT and 324 myocardial segments of HT controls were analysed and compared using linear mixed modeling. Mean interventricular septal thickness and left ventricular mass indexed to body surface area was similar between the groups . None of the advanced CKD group had impaired global longitudinal strain (GLS) (mean GLS -18.39) and none of the RT/HT groups had late gadolinium hyperenhancement. The mean BOLD SI change was lower in advanced CKD and RT groups compared to HT controls . The global myocardial BOLD SI change was also lower in the advanced CKD subjects compared to RT recipients (p=0.045). In the advanced CKD and RT groups, the BOLD SI Change was associated with eGFR .Our study suggests myocardial oxygenation is impaired in advanced chronic kidney disease patients and renal transplant recipients, and unlikely to be related to LVH or myocardial scarring. The impaired myocardial oxygenation may be associated with declining renal function. Non-contrast BOLD CMR is a promising tool to detect myocardial ischaemia in advanced chronic kidney disease population.None."} +{"text": "PTPN22 gene locus results in aberrant function of PTPN22 protein and protects from Crohn\u2019s disease (CD). Here, we investigated associations of PTPN22 SNP rs2476601 in inflammatory bowel disease (IBD) patients in the Swiss IBD Cohort Study (SIBDCS).Protein tyrosine phosphatase non-receptor type 22 (PTPN22) plays an important role in immune cell function and intestinal homeostasis. The single nucleotide polymorphism (SNP) rs2476601 within the 2\u2019028 SIBDCS patients (1173 CD and 855 ulcerative colitis (UC) patients) were included. The clinical characteristics were analysed for an association with the presence of the PTPN22 SNP rs2476601 genotypes \u2018homozygous variant\u2019 (AA), \u2018heterozygous\u2019 (GA) and \u2018homozygous wild-type\u2019 (GG).PTPN22 variant featured malabsorption syndrome (p = 0.026).13 patients (0.6%) were homozygous variant (AA) for the PTPN22 polymorphism, 269 (13.3%) heterozygous variant (GA) and 1\u2019746 (86.1%) homozygous wild-type (GG). In CD, AA and GA genotypes were associated with less use of steroids and antibiotics, and reduced prevalence of vitamin D and calcium deficiency. In UC the AA and GA genotype was associated with increased use of azathioprine and anti-TNF antibodies, but significantly less patients with the Our study for the first time addressed how presence of SNP rs2476601 within the PTPN22 gene affects clinical characteristics in IBD-patients. Several factors that correlate with more severe disease were found to be less common in CD patients carrying the A-allele, pointing towards a protective role for this variant in affected CD patients. In UC patients however, we found the opposite trend, suggesting a disease-promoting effect of the A-allele. A single nucleotide polymorphism (SNP) within the gene locus encoding protein tyrosine phosphatase non-receptor type 22 has been associated with an increased risk to develop autoimmune disorders, including rheumatoid arthritis (RA), systemiPTPN22 gene locus and results in the substitution of arginine 620 with a tryptophan residue in the PTPN22 protein product (PTPN22-620W). Although initial studies demonstrated that presence of the variant results in increased in vitro dephosphorylation capacity2 test or the Fisher's exact test (Fisher's exact test used if strata comprised a sample size \u22645). A multiple logistic regression model was calculated to identify the associations for this gene variant. Differences about the association of the PTPN22 variant in relation to age at diagnosis were assessed using a Wilcoxon rank-sum test. A p-value smaller than 0.05 was considered significant.Crude differences about the association of the The Swiss IBD cohort study is approved by the local ethical committees . Written informed consent was obtained before inclusion in the cohort.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file."} +{"text": "Staphylococci are one of the foremost causes of urinary tract infections (UTIs) in humans. The emergence of multiple drug resistance (MDR) among Staphylococci poses serious challenges in antimicrobial therapy for UTIs. Most work has been done on S. aureus while coagulase negative Staphylococci (mainly S. epidermidis) are often neglected. This study was conducted to establish a baseline profile of drug resistance in local S. epidermidis isolates from UTIs. Eighty urine samples were collected from suspected UTIs cases and screened for S. epidermidis. Twenty isolates were suspected as S. epidermidis based on colony morphology and Gram staining. Molecular detection by polymerase chain reaction (PCR) confirmed 13 isolates as S. epidermidis. Using disc diffusion method, phenotypic drug resistance of the isolates was observed towards erythromycin (100 %), gentamycin, azithromycin and tetracycline (92.3 %), ampicillin and oxytetracyclin (84.6 %), amikacin and srteptomycin (76.9 %), methicillin (69.2 %), cephradine, cefaclor and cefazolin (53.8 %) and vancomycin (15.3 %). Eighteen most commonly reported genes responsible for conferring resistance towards these drugs were targeted by PCR: among these tetM gene was found most prevalent (46.1 %) followed by tetK (30.7 %), aac(6')/aph(2\u201d) (30.7 %), aacA-aphD (23 %), ermA (23 %), blaZ (23 %), mecA (23 %) blaTEM-1 (23 %), MeccA (23 %) and mecA (15.3 %). No gene fragment for vancomycin resistance was detected. The salient finding was that all S. epidermidis isolates were multiple drugs resistant as they showed resistance against at least three structurally different antimicrobial agents. It is concluded that in addition to the mostly used antimicrobial agent vancomycin, the cephalosporins including cephradine, cefaclor and cefazolin are also the drugs of choice against UTIs caused by S. epidermidis. The UTild, 2007). There ild, 2007). UTIs iild, 2007).The risk of UTIs is more in females; one reason is the shorter urethra in women that allows bacteria quicker access to reach the bladder . RecurrEscherichia coli is reported as the most common bacterium and it accounts for about 85 % of community-acquired and 50 % of hospital acquired urinary tract infections followed by Staphylococcus species that contribute up to 15 % .Staphylococcus epidermidis is a coagulase negative Staphylococci and one of the most often reported Staphylococcus in nosocomial infections. It usually enters the urinary tract through urethra and resides on the skin or mucous membrane was found resistant to more antibiotics than S. epidermidis colonizing healthy volunteers. It clearly suggests the S. epidermidis as potential cause of catheter related infections due to its antimicrobial resistance based adaptability to survive in the hospital environment . To theStaphylococcus colonies. The suspected isolates were sub-cultured on the same medium for obtaining pure cultures and processed for biochemical and molecular identification.Urine samples (no = 80) were aseptically collected from patients of all age groups and both sexes. The samples were collected from different laboratories and hospitals of Faisalabad. Sterile leak resistant containers were used for collection and transportation of urine from hospitals to our laboratory and inoculated into 0.6 % nutrient agar in tryptic soya broth. The well-mixed unspun urine was streaked on nutrient ager plates using sterile swab and incubated overnight at 37 \u00b0C. The agar plates were observed for the presence of Staphylococcus were processed for Gram-staining and observed under light microscope on glasStaphylococci was extracted from the overnight culture at 37 \u00b0C in tryptic soy broth (TSB) using the phenol-chloroform method , followed by 35 cycles of 95 \u00b0C for 1 min, 60 \u00b0C for 40 sec, 72 \u00b0C for 50 sec and a final extension of 72 \u00b0C for 7 min.Total genomic DNA from all isolates suspected as al., 1989). The inal., 2000) targetiAfter PCR confirmation of the isolates, antimicrobial susceptibility testing was performed by disc diffusion method against six different antimicrobial groups, using discs of thirteen representative antimicrobials: amikacin (30 \u03bcg), ampicillin (10 \u03bcg), cefradine (30 \u03bcg), cefaclor (30 \u03bcg), cefazolin (30 \u03bcg), gentamicin (10 \u03bcg), streptomycin (10 \u03bcg), vancomycin (30 \u03bcg), azithromycin 15 \u03bcg), erythromycin (15 \u03bcg), methicillin (10 \u03bcg), tetracycline (30 \u03bcg) and oxytetracycline (30 \u03bcg). The diameter of zone of inhibition, if present was measured and results for all antimicrobials used were interpreted according to the guidelines of Clinical And Laboratory Standard Institute the resistance genes (aac(6')/aph(2\u201d) were taal., 2000) for cepal., 2003). For teal., 2003). For beenes (aac'/aph(2\u201d)al., 1995). Seven Staphylococci based on colony morphology and Gram staining (positive). All these isolates were found coagulase negative as no clumping of human plasma was observed with them while the positive control Staphylococcus aureus showed clear clumping so all the isolates were considered as multiple drug resistant (MDR) isolates . With daacA-aphD gene in 3 isolates (23 %), erm(A) gene fragment in 3 isolates (23 %), tetK gene fragment (360 bp) in 4 isolates (30.7 %) and tetM in 6 (46.1 %) isolates. The blaZ gene which confers resistance to penicillin was detected in 3 isolates (23 %). No amplification was observed with primer sets used for genes reported to confer resistance against vancomycin. In the duplex PCR for MeccA and mecA, responsible for conferring resistance to methicillin, the MeccA was amplified in 3 isolates (23 %) and mecA was amplified in 2 isolates (15.3 %) while in 3 phenotypically resistant isolates, no gene was detected for methicillin resistance. The gene fragment of bla TEM-1 was amplified (962 bp) in 3 isolates (23 %) while the gene aac(6')/aph(2\u201d) responsible to confer resistance to gentamicin was found (1184 bp) in 4 isolates (30.7 %). The representative amplified products are shown in Figure 3The PCR based detection of different antimicrobial resistance genes showed the presence of S. epidermidis in different ways, as each antimicrobial drug has a specific mode of action are usually caused by pathogenic microorganisms . These are considered the second common category of infections in the body . The urNeu, 1992). Staphylococci has been reported to be associated with the presence of PBPs encoded by the MeccA gene , were identified in three and two isolates respectively. The possible reason for this difference in genotypic and phenotypic antimicrobial sensitivity patterns is the presence of other drug resistance genes which were not included in this study. Results about resistance to methicillin found in this study were higher than those reported (34.2 %) from Turkey of bacteria also have resulted in reduced affinity for \u00df-lactams . Methical., 2008; Choi etal., 2008). The hial., 2007) and Eural., 2007) but weral., 2007).aac(6')/aph(2\u201d) and aacA-aphD gene fragments was also checked. The gene aac(6')/aph(2\u201d) responsible to confer resistance against gentamicin was amplified (1184 bp) in 4 isolates (30.7 %) and aacA-aphD was amplified (227 bp) in 3 isolate (23 %). In accordance with previous studies /aph(2'') gene as the most prevalent AME gene in Staphylococci. Contrarily, a study from Japan reported less frequent detection of aac(6')/aph(2'') gene among clinical MRSA isolates isolates in our study by disc diffusion method. The presence of aminoglycoside-modifying enzyme (AME) genes al., 1994; Choi etal., 2001).erm(A) gene fragment in only 3 isolates (23 %) while no amplification was observed for erm(C) gene fragment. Previously, the erm(A)gene was reported as more prevalent than the other erythromycin resistance genes in S. aureus while erm(C) gene is present more frequently in coagulase negative Staphylococci isolates while tetM gene fragment was amplified in 6 (46.1 %) isolates. However, we found neither of these genes in 4 (30.7 %) phenotypically resistant isolates which indicates the possibility of carrying some other tetracycline resistance genes isolates were found phenotypically resistant while PCR assay detection lum, 1995) which wrts, 2005) or harbrts, 2005).vanR, vanS, vanH, vanA, vanX, vanYand vanZ. One possible reason is that the most effective form of vancomycin resistance depends on a transposon containing seven genes, the products of which work together to sense the presence of vancomycin, shut down the normal pathway for bacterial cell wall synthesis, and generate a different type of cell wall. Therefore, the joining of these genes into a single transposon must have been a difficult evolutionary step. Moreover the bacteria may have some other alternative strategies to confer resistance against vancomycin. There are only a few cases reported all over the world about isolation of vancomycin resistant S. aureus (VRSA) from clinical specimens and among these isolates small number was community-acquired. The first case of community-acquired methicillin and vancomycin-resistant S. aureus was reported in Tehran, Iran. According to it in-vitro transfer of vancomycin resistance gene (vanA) from the source Enterococci to S. aureus isolates, it was suspected that there is a possibility of transformation of vanA gene from vancomycin resistant Enterococci (VRE) to Staphylococci species isolates resistant to vancomycin by disc diffusion method but not a single isolate showed amplification in the multiplex PCR for the targeted seven gene fragments i.e. al., 2004).S. aureus (MRSA) to several antimicrobial agents has aggravated due to the over use of vancomycin as a first-line empirical therapy as well as in prophylaxis therapy, though the selection of vancomycin resistance and the potential transmission of resistance in species encourage restricted use of these glycopeptides .bla TEM-1 gene involved in conferring resistance to cephalosporins was found in only 3 isolates (23 %). We conclude that vancomycin is the drug of choice against UTIs caused by Staphylococci and cephalosporins including cephradine, cefaclor and cefazolin are also still effective in this geographic region.Another major group of antimicrobial drugs is cephalosporins. These drugs are very efficient and several generations are available in the market. First generation preparations are generally active against gram-positive bacteria. We used three drugs i.e. cephazolin, cefradine and cefaclor. With disc diffusion method, 7 (53.8 %) isolates were observed to be resistant against these antimicrobials. While in a multiplex PCR assay"} +{"text": "Adenosine stress magnetic resonance is highly accurate for the detection of myocardial perfusion abnormalities. Visual assessment of inducible wall motion abnormalities (IWMA) during adenosine has low sensitivity but high specificity since only high-grade perfusion defects are associated with detectable IWMAs. The novel technique Feature Tracking (FT) measures myocardial strain and gives detailed information about regional myocardial deformation. The aim of this study was to investigate FT for the detection of myocardial ischemia during adenosine stress.A total of 62 patients with suspected or known coronary artery disease (CAD) underwent adenosine stress CMR at 1.5 or 3T. Patients with evidence of myocardial scar iby late gadolinium enhancement (LGE), WMA at rest and impaired left ventricular function (LVEF) were excluded (n = 34). Short axis (SAX) cine views were acquired before and after 3 minutes of continuous adenosine infusion (0.14 mg/min/kg). Patients with abnormal stress perfusion were considered positive for myocardial ischemia (n = 15) and segments with perfusion defects were defined as ischemic (n = 61). Patients without perfusion defects and without known CAD were defined as normal and served as control . TomTec 2D Cardiac Performance Analysis software was used to derive quantitative assessment of circumferential strain (CS) and strain rate (SR) from the three SAX cine views at rest and under stress.There was no significant difference of CS at rest between normal and ischemic segments . Normal segments demonstrated a significant increase during stress for CS as well as for SR . In ischemic segments no significant difference was observed for CS between rest and stress (-28.8 \u00b1 9.6% vs. 31.9 \u00b1 12.8%, p = 0.064; Figure Strain analysis with FT during adenosine stress demonstrated significant differences between normal and ischemic segments. Therefore it may be a valuable additional tool for the assessment of myocardial ischemia.No funding."} +{"text": "Histopathologic changes of the SM may include fibrosis and fatty degeneration2. The aim of this study is to evaluate myocardial structure in preserved ejection fraction (EF).Myotonic dystrophy type II (MD2) is a genetic multisystemic disorder characterized by skeletal muscle (SM) symptoms, metabolic changes as well as arrhythmias3 was performed to identify myocardial fat deposits. Furthermore, we used 1H magnetic resonance spectroscopy (MRS) to quantify myocardial Triglycerides (MTG). Data were analyzed using cvi42 and standard line-fitting procedure.We prospectively enrolled 32 subjects with a genetically confirmed diagnosis of MD2. Exclusion criteria were known cardiac diseases and contraindication for CMR. We assessed left-ventricular (LV) volumes, mass and function applying state of the art cine imaging using a 1.5 T Scanner. Late enhancement imaging 7 mm) was performed to detect myocardial fibrosis 10 minutes after injection of gadoteridol (0.2 mmol/kgbw). We applied T1 Mapping based on MOLLI before and 15 minutes after contrast application and assessed resultant extracellular volume fraction (ECV). Fat-water-separated imaging26 data sets were totally completed . None of the patients had wall motion abnormalities. LGE was detectable in 6 of 28 subjects ; the location was mostly subepicardial inferolateral basal (Figure Despite preserved LVEF we could detect myocardial injury in patients with myotonic dystrophy type II. Already native T1-values were increased in case of subepicardial fibrosis compared to remote myocardium. To conclude, this is the first study constituted that CMR is feasible to detect subclinical myocardial manifestations in MD2.N/A."} +{"text": "Hand hygiene compliance is poor among graduated nurses. Hand hygiene behaviour is complex and shows great variation that can be attributed to the healthcare worker\u2019s educational background.The objective was to quantify the level of attention given to hand hygiene in the course curricula of nursing schools in Belgium.A questionnaire was distributed during the Federal Public Service\u2019s (FPS) \u2018hand hygiene campaign introduction session 2011\u2019 for nursing school staff involved in teaching hand hygiene.Response was 100% (n=49). The attending teachers represented 53% of all Belgian nursing schools. With each year of education, HH related knowledge , attitude and skills receive less attention. Competencies are evaluated during clinical practice (90%), by means of written exams (90%), skills tests at school (37%) or \u2018other\u2019 (18%). Most popular teaching methods include live demonstration (98%), skills training sessions (71%) and video demonstration (59%). Courses are being held up to date using books (64%), publications of the Superior Health Council (44%), scientific publications (26%) and the FPS HH website (16%). Sixty-one per cent of the study books have not been updated in the past two years. The average time spent on infection control and hand hygiene was respectively 19.68hrs and 7.07hrs .Hand hygiene education in Belgium shows great variation. Hand hygiene competencies are not systematically acquired nor assessed during the 3-year course. Moreover, teaching and assessment methods show considerable room for innovation.None declared."} +{"text": "Imaging appearance of primary lung tumour conditions that on initial radiological studies might be confused with malignant lesions.Diagnostic tools for evaluation of thoracic tumours and pseudotumours. We included in the differential diagnosis:Paraffinoma (1), a hydatid cyst (1), inflammatory pseudotumour (3), nodular tuberculosis (2) and sarcoidosis granulomas (1), round pneumonia (1), nodular criptococosis (1), post-surgical or tuberculosis scarring processes (2), organising cryptogenetic pneumonia (2) and round atelectasis (3).There are several diseases that can mimic tumours of the chest. It is necessary to take them into account and know how to use diagnostic keys for an accurate diagnosis."} +{"text": "On the other hand, our results showed that Tdp2 gene knockout did not block CCC DNA formation during HBV infection of permissive human hepatoma cells and did not prevent intracellular amplification of duck hepatitis B virus CCC DNA. These results indicate that although Tdp2 can remove the RT covalently linked to the 5\u2019 end of the HBV minus strand DNA in vitro, this protein might not be required for CCC DNA formation in vivo.Hepatitis B virus (HBV) replication and persistence are sustained by a nuclear episome, the covalently closed circular (CCC) DNA, which serves as the transcriptional template for all viral RNAs. CCC DNA is converted from a relaxed circular (RC) DNA in the virion early during infection as well as from RC DNA in intracellular progeny nucleocapsids via an intracellular amplification pathway. Current antiviral therapies suppress viral replication but cannot eliminate CCC DNA. Thus, persistence of CCC DNA remains an obstacle toward curing chronic HBV infection. Unfortunately, very little is known about how CCC DNA is formed. CCC DNA formation requires removal of the virally encoded reverse transcriptase (RT) protein from the 5\u2019 end of the minus strand of RC DNA. Tyrosyl DNA phosphodiesterase-2 (Tdp2) was recently identified as the enzyme responsible for cleavage of tyrosyl-5\u2019 DNA linkages formed between topoisomerase II and cellular DNA. Because the RT-DNA linkage is also a 5\u2019 DNA-phosphotyrosyl bond, it has been hypothesized that Tdp2 might be one of several elusive host factors required for CCC DNA formation. Therefore, we examined the role of Tdp2 in RC DNA deproteination and CCC DNA formation. We demonstrated Tdp2 can cleave the tyrosyl-minus strand DNA linkage using authentic HBV RC DNA isolated from nucleocapsids and using RT covalently linked to short minus strand DNA produced Hepadnaviridae family of viruses, which include the duck hepatitis virus (DHBV). All hepadnaviruses contain a small (ca. 3.2 kb) relaxed circular (RC), partially double-stranded DNA genome that replicates via reverse transcription through an RNA intermediate called pregenomic RNA (pgRNA) -TTP. The DNA products of this reaction were resolved by urea- PAGE, and visualized by autoradiography. The autoradiograph showed the two expected DNA products from the Sfc I digestion and Klenow fill-in reaction of 43 and 182 nt derived from the minus and plus strand DNA, respectively (in vitro.A recent study showed that pretreatment of the HBV and DHBV RC DNA\u2013RT complex released from NCs with Tdp2 could make the 5\u2019 end of the (-) strand susceptible to 5\u2019 exonuclease digestion, suggesting that Tdp2 could release the RT protein from RC DNA . To direin vitro protein priming reaction to produce RT proteins covalently attached to a short DNA oligomer several nt long via the phosphotyrosyl bond -TTP for 2 h at 25\u00b0C using Klenow (NEB). RNase digestion was used to remove the RNA primer attached to the 5\u2019 end of the plus strand DNA. The digested DNA was extracted as above by phenol/chloroform extraction and pellet paint precipitation. Pellets were resuspended in 2X formamide loading buffer (Ambion) and boiled for 5 min prior to resolve by urea PAGE. All reaction products were visualized by autoradiography.Full-length human Tdp2 recombinant protein with glutathione 32P]-dGTP and the other three unlabeled dNTPs, and TMnNK buffer . For trans-complementation priming assays [in vitro protein priming products were carried out as previously described [DHBV protein priming assays were carried out as previously described \u201328. Brieg assays , purifieescribed , 24. SDS"} +{"text": "Nbeal2 gene was identified. Though initially considered as a plausible thrombosis modifier, this Nbeal2 mutation failed to suppress the synthetic lethal thrombosis on which the original ENU screen was based. Mutations in NBEAL2 cause Gray Platelet Syndrome (GPS), an autosomal recessive bleeding disorder characterized by macrothrombocytopenia and gray-appearing platelets due to lack of platelet alpha granules. Mice homozygous for the Nbeal2 8 bp deletion exhibit a phenotype similar to human GPS, with significantly reduced platelet counts compared to littermate controls (p = 1.63 x 10\u22127). Nbeal2gps/gps mice also have markedly reduced numbers of platelet alpha granules and an increased level of emperipolesis, consistent with previously characterized mice carrying targeted Nbeal2 null alleles. These findings confirm previous reports, provide an additional mouse model for GPS, and highlight the potentially confounding effect of background spontaneous mutation events in well-characterized mouse strains.During the analysis of a whole genome ENU mutagenesis screen for thrombosis modifiers, a spontaneous 8 base pair (bp) deletion causing a frameshift in exon 27 of the These inbred strains have been extensively characterized and the genome of more than 20 have been sequenced , 2. Whols (SNVs) , each hus (SNVs) , 5. Mices (SNVs) and thers (SNVs) . However studies \u201311. Forwutations , 13. ENUutations , 15.NBEAL2 encodes neurobeachin-like-2, a BEACH domain containing protein, with a proposed role in vesicular trafficking and granule development [NBEAL2 were recently shown to be the cause of the autosomal recessive form of Gray Platelet Syndrome (GPS) [Nbeal2 [elopment . Mutatiome (GPS) \u201319. GPS me (GPS) . Mice wi [Nbeal2 \u201323 exhib [Nbeal2 , 24.Nbeal2 gene. Analysis of the associated mouse pedigree demonstrated that this mutation arose within the Jackson laboratory 129S1/SvImJ mouse colony and not from the ENU screen.During the analysis of a whole genome ENU mutagenesis screen for thrombosis modifiers, we identified a spontaneous 8 bp deletion causing a frameshift in exon 27 of the Animal husbandry in this study was carried out according to the Principles of Laboratory and Animal Care established by the National Society for Medical Research. The University of Michigan\u2019s University Committee on Use and Care of Animals (UCUCA) has approved the protocol number 05191 and the University of Colorado Institutional Animal Care and Use Committee approved the protocol 96114. The care and maintenance of animals was closely supervised by University of Michigan ULAM personnel or University of Colorado Institutional Animal Care and Use Committee (IACUC) and animals were housed in their facilities. ULAM/IACUC also provided expert veterinary advice and assistance when necessary and cages were monitored closely by our laboratory personnel as well as university veterinary staff. To minimize discomfort and unnecessary suffering of experimental mice, analgesics were administered for all procedures involving significant discomfort. Blood samples were obtained from the retro-orbital plexus of anesthetized animals achieved with isoflurane inhalation. Mice were euthanized for collection of tissues for histologic, biochemical, and genetic analysis. The UCUCA Endstage Illness and Humane Endpoint Guidelines were also closely followed and animals euthanized accordingly by carbon dioxide overdose or exsanguination under anesthesia.F5L/L (F5tm2Dgi/J stock number 004080) mice were previously generated [Tfpi deficient mice (Tfpitm1Gjb) were a generous gift of Dr. George Broze [Nbeal2tm1Lex/tm1Lex mice with targeted deletion of the Nbeal2 gene were previously generated from cryopreserved spermatozoa obtained from the Mutant Mouse Regional Resource Center at the University of California, Davis [Nbeal2 allele carrying the spontaneous 8bp deletion described in Results will be referred throughout the text as Nbeal2gps. Two cohorts of Nbeal2gps mice were analyzed. Set 1 refers to Nbeal2gps mice intercrossed after 2 backcrosses to C57BL/6J mice (stock number 000664), while set 2 mice were intercrossed after 7 backcrosses to C57BL/6J.enerated , Tfpi dege Broze , and Nbea, Davis . Nbeal2 Mus Musculus GRCm38 reference genome, duplicates were removed using Picard [MF5L6 with a minimum of 6X coverage were considered as potential candidates and further validated using Sanger sequencing.Genomic DNA (gDNA) was extracted from mouse tail biopsies using the Gentra Puregene Tissue Kit (Qiagen) according to manufacturer\u2019s instructions. Exonic DNA was captured with either SureSelect Mouse All Exon (Agilent) or SeqCap EZ Mouse Exome Design (NimbleGen) kits and 100 bp paired-end sequencing was performed on the Illumina HiSeq 2000 platform at the University of Michigan's DNA Sequencing Core. All generated fastq files have been deposited to the NCBI Sequence Read Archive (Project accession number #SRP063933). Detailed overview of the variant calling pipeline and filtration is available online as a GitHub repository . In shorg Picard , and varg Picard . Variantg Picard with RefExome analysis was performed for the parents (F63pF64) and a female sibling (F63pF65) of the 129S1/SvImJ individual sequenced for the Sanger Mouse Genomes Sequencing project . ApproxiNbeal2gps allele was detected using two three-primer PCR assays is located over the undeleted 8 bp to detect the presence of the wildtype allele. This PCR reaction results in two products . In the second PCR design the third primer spans the 8 bp deletion to detect the presence of the deletion allele. This reaction also produces two products . PCR was performed using GoTaq Green Master Mix (Promega) and the products visualized on a 2% agarose gel. Selected genotyping results were further confirmed by Sanger sequencing.The R assays with comNbeal2gps/+ mouse. Total RNA was extracted using an RNeasy Mini Kit (Qiagen) and converted to coding DNA (cDNA) using SuperScript III One-Step RT-PCR (Invitrogen) following the manufacturer\u2019s instructions. gDNA was prepared from a tail biopsy. Forward and reverse genotyping primers (primer F and primer R) were used to amplify the Nbeal2 deletion region from gDNA and the cDNAs from liver and lung. PCR products were extracted from agarose gels using a QIAquick Gel Purification Kit (Qiagen) and submitted for Sanger sequencing. The differential allelic expression was estimated from the ratio between the wildtype and Nbeal2gps sequence peak areas in cDNA samples compared to gDNA using Phred software [Nbeal2gps alleles contain a different nucleotide.Liver, lung, and bone marrow tissue samples were collected in RNAlater (Ambion) from a software . This ra2HPO4, 2 mM KCl, 12 mM NaHCO3, 5 mM HEPES, 5 mM glucose) [Murine whole blood was collected via the inferior vena cava into acid/citrate/dextrose. Platelet-rich plasma (PRP) was obtained by centrifugation at 200 g for 5 min. Washed platelets were pelleted from PRP by centrifugation at 1,000 g for 2 min in the presence of prostacyclin PGI1 (0.1 \u03bcM) and resuspended in modified Tyrode\u2019s buffer . Total pTwenty-five microliters of blood were collected from the retro-orbital sinus of 5\u20136 week old mice from set 1. Blood was anticoagulated with 4% sodium citrate (Sigma-Aldrich) and diluted 10x in PBS supplemented with 5% bovine serum albumin (Sigma-Aldrich). Complete blood counts (CBC) were performed on the ADVIA 2120 Hematology System (Siemens) according to manufacturer\u2019s instructions while being blinded to the genotype of the mouse from which the sample was obtained. Additional blood was collected from >20 week old females from set 2 using heparinized capillary tubes and anticoagulated using EDTA containing tubes (BD microtainer). CBC were performed on the Hemavet 950FS system. All data were analyzed and visualized using the stats and beeswarm packages in R software .Absolute neutrophil counts (ANC) were measured by flow cytometry as previously described . BrieflyPeripheral blood smears were prepared from 9 mice of each genotype and Wright-Giemsa stained using the HealthCare PROTOCOL Hema 3 kit according to the manufacturer\u2019s instructions (Fisher Scientific). For each sample, the intensity of platelet staining and platelet granularity were categorized into three levels by one of the authors (RK) blinded to the genotype of the mouse from which the sample was obtained. Representative images from the blood smears were taken using a Leica DMLB microscope at 1000x magnification. Bone marrow sections as well as bone marrow cytology slides were prepared by the Unit for Laboratory Animal Medicine histology core. Histopathologic evaluation was performed by an investigator blinded to the genotypes of the evaluated mice.Nbeal2+/+ and one Nbeal2gps/gps mouse from set 1 was collected by retro-orbital puncture and fixed in 4% glutaraldehyde as previously described [Blood from one escribed . Fixed sescribed for 100 Nbeal2gps/gps and wildtype mice. A chi-square test was applied to estimate deviations from expected Mendelian proportions in Nbeal2 mouse crosses. All statistical analyses were performed using the stats package in R software [A non-parametric Wilcoxon test was used to estimate significance in the CBC measured values, platelet area, the assigned platelet staining intensity values of the Wright-Giemsa stained blood smears, and the difference in the level of emperipolesis in bone marrow slides between software .F5L/LTfpi+/-) in C57BL/6J mice [F5L/L mice [F5L/LTfpi+/- mice. Whole exome sequencing was applied to 4 mice from one of the suppressor lines ENU mutagenized progeny exhibiting the suppressor phenotype colony at The Jackson Laboratory were genotyped, including the 129S1/SvImJ founder pair, \u201cAdam and Eve\u201d (F60) as well as archived samples from before and after implementation of the GSP program. The Nbeal2 deletion was not found in any of these samples . In this samples . Exome s or two deletion alleles were viable, fertile and had no apparent phenotype by visual inspection. No significant deviation from the expected Mendelian distribution was observed in the progeny when crossing the Nbeal2gps/+ mice to C57BL/6J wildtype mice or in the progeny from the Nbeal2gps/+ intercross as was the absolute neutrophil count and mean platelet volume (p = 0.016) were higher in the Nbeal2gps/gps mice compared to littermate controls (Complete blood counts (CBC) were performed on 24 rameters and thos x 10\u22129) , Fig 2. rol mice . In addiNbeal2gps/+ and wildtype mice , but significantly reduced in Nbeal2gps/gps mice (-4) consistent with a reduction in platelet alpha granules [Nbeal2gps/gps mouse compared to wildtype control in the bone marrow compared to wildtype mice, consistent with previously reported human and mouse GPS phenotypes [Nbeal2gps/gps mice exhibited some degree of emperipolesis (\u22128). Megakaryocytes containing more than one neutrophil were observed exclusively in the bone marrow of Nbeal2gps/gps mice. Similarly, increased emperipolesis was observed in spleens of Nbeal2gps/gps mice demonstrated differences in neutrophil counts and mean platelet volumes .Our data establish that the enotypes . More reory mice , which iNDEL see . This isNbeal2gps allele was identified via whole exome sequencing of the progeny of an ENU treated mouse. While next generation sequencing approaches have high utility for mapping both spontaneous [de novo variants [The ntaneous as well variants , the orivariants .de novo variants are easily detected in mouse colonies; however, mild dominant phenotypes or recessive phenotypes may go unnoticed depending on the breeding paradigm. For these reasons, it is important to adhere to published guidelines on mouse colony management and genetic quality control monitoring [Nbeal2gps allele, the platelet defect had no impact on survival of F5L/LTfpi+/- mice and we were able to identify the variant only due to next generation sequencing.Ultimately, the origin of causative mutations (ENU or spontaneous) can be established through additional genotyping of the ENU pedigree, assuming breeding records and samples have been carefully maintained and archived. Generally, strong dominant phenotypes due to nitoring . In the S1 Fig(PNG)Click here for additional data file.S2 FigF5L/LTfpi+/- genotype and unaffected parents are shown in the pedigree. Black boxes highlight the mice subjected to whole exome sequencing. The red box highlights mouse 67339 that was used for Nbeal2gps allele outcrossing and line establishment.Only progeny mice with the (PDF)Click here for additional data file.S3 FigNbeal2gps/+ progeny (red). One of these progeny (asterisk) was the sire of the female used to build the ENU suppressor line (A). All genotyped 129S1/SvImJ (JR# 002448) mice were wildtype at the Nbeal2 locus, including the \u201cAdam and Eve\u201d founders of The Jackson Laboratory GSP 129S1/SvImJ stock (F60) [Nbeal2 deletion was also absent in two post-GSP 129S1/SvImJ animals: whole exome sequencing data (F63pF67) from the Mouse Mutant Resource [Two different mice A91, A92) purchased from The Jackson Laboratory had ck (F60) and two Resource and whol, A92 pur(PDF)Click here for additional data file.S4 FigNbeal2gps allele is lower than wildtype, set as 100% (A). Boxplot of all data points show on average ~65% reduction in expression of the deletion allele (B).Allelic expression was measured at every position in the Sanger sequenced RT-PCR product where the reference and deletion alleles had a different nucleotide. Dotted lines fill the gaps. In all tested tissues, the relative expression of (PDF)Click here for additional data file.S5 FigHere we show full blots for the Western blot analysis. Areas surrounded by black boxes were displayed in (PDF)Click here for additional data file."} +{"text": "The primary goal of this study was to evaluate the incidence and characteristics of post-traumatic headache(PTH) attributed to mild brain injury in military personnel in Iran within a prospective and observational study design.A prospective observational study was conducted with a cohort of military personnel under military education during 6 months period at Amiralmomenin Military education center in Isfahan in Iran. Through all military personnel under education 322 personnel were selected randomly in simple manner were given 13-item Mild brain injury questionnaire accompany with affective disorders and headache questionnaires and were reevaluated in 3 months interval.A total of 30 (9.3%) of 322 military personnel met criteria for a mild brain injury. Among them 18 personnel (60%) reported having headaches during 3-month of reevaluation. Patients with affective disorders such as post-traumatic stress disorder (PTSD) and depression were at higher risk for developing PTH following mild brain injury (p<0.05). PTH did not relate to demographic factors such as age or type of trauma. At follow up, PTH had abated in 3 months except in 2 patients developed chronic PTH pattern. We did not observe any significant difference of medications prescribed by practitioners between different classified patterns of PTH (p>0.05).PTH attributed to mild brain injury is a common disorder in military personnel. Migrainous features are predominant among them in comparison with general population. PTH is not related to type of trauma but has association with affective disorders.No conflict of interest."} +{"text": "Bronchopulmonary Dysplasia (BPD) is a major complication of preterm birth associated with significant morbidity. BPD is a debilitating condition characterized by inflammation, enlarged airspaces, vascular dysmorphia and aberrant extracellular matrix accumulation that is typically described as arrested lung development. Rodent models involving neonatal exposure to excessive oxygen concentrations (hyperoxia) have been used to study the mechanisms contributing to BPD pathology. Transcriptomic assessment of the effects of hyperoxia in neonatal mouse lungs using RNASeq will help to identify genes and pathways associated with BPD.Whole lung tissue from newborn C57BL/6 mice exposed to 100% oxygen for 10 days (n=8) and room air-exposed age matched controls (n=6) were compared. Total RNA was isolated from individual whole lung tissues (n=14) and pooled in duplicates to perform transcriptome Sequencing (RNA-seq). Alignments were generated using multiple algorithms . Raw counts obtained from each alignment algorithm (using HT-Seq) were further and filtered to remove undetected genes. Differentially expressed genes were detected using Significance Analysis of Microarrays (SAM) and CuffDiff2, on each version of mapped and normalized data. Ingenuity Pathway Analysis (IPA) was used for pathway and network analyses. Expression patterns for selected genes were examined by quantitative polymerase chain reaction (qPCR).248 genes were identified as differentially expressed between hyperoxia and control samples by both SAM (median FDR = 0) and CuffDiff2 (p<0.05) and had a fold-change \u2264 2. We successfully validated 17 of 24 genes by qPCR. Canonical pathways significantly dysregulated in hyperoxia lungs included Nrf2-mediated oxidative stress signaling, p53 signaling, hepatic fibrosis and sildenafil pathways. Interestingly most genes significantly affected following hyperoxia exposure (~70%) showed a pattern of expression consistent with an arrest in lung development. A subset of the genes dysregulated in hyperoxic neonatal mouse lungs, were also differentially expressed in human BPD lung tissue.We have identified genes dysregulated in mouse of BPD-like pathology. Further analysis of these data will enhance our current knowledge of BPD, and may be useful for developing novel therapeutic strategies."} +{"text": "PRNP). A proline to leucine substitution at PrP residue 102 (P102L) is classically associated with Gerstmann-Str\u00e4ussler-Scheinker (GSS) disease but shows marked clinical and neuropathological variability within kindreds that may be caused by variable propagation of distinct prion strains generated from either PrP 102L or wild type PrP. To-date the transmission properties of prions propagated in P102L patients remain ill-defined. Multiple mouse models of GSS have focused on mutating the corresponding residue of murine PrP (P101L), however murine PrP 101L, a novel PrP primary structure, may not have the repertoire of pathogenic prion conformations necessary to accurately model the human disease. Here we describe the transmission properties of prions generated in human PrP 102L expressing transgenic mice that were generated after primary challenge with ex vivo human GSS P102L or classical CJD prions. We show that distinct strains of prions were generated in these mice dependent upon source of the inoculum (either GSS P102L or CJD brain) and have designated these GSS-102L and CJD-102L prions, respectively. GSS-102L prions have transmission properties distinct from all prion strains seen in sporadic and acquired human prion disease. Significantly, GSS-102L prions appear incapable of transmitting disease to conventional mice expressing wild type mouse PrP, which contrasts strikingly with the reported transmission properties of prions generated in GSS P102L-challenged mice expressing mouse PrP 101L. We conclude that future transgenic modeling of IPDs should focus exclusively on expression of mutant human PrP, as other approaches may generate novel experimental prion strains that are unrelated to human disease.Inherited prion disease (IPD) is caused by autosomal-dominant pathogenic mutations in the human prion protein (PrP) gene ( Inherited prion disease (IPD) is caused by pathogenic mutations in the human prion protein (PrP) gene leading to the formation of lethal prions in the brain. To-date the properties of prions causing IPD and their similarities to prions causing other forms of human prion disease remain ill-defined. In the present study we have investigated the properties of prions seen in patients with Gerstmann-Str\u00e4ussler-Scheinker (GSS) disease associated with the substitution of leucine for proline at amino acid position 102 (GSS P102L). We examined the ability of these prions to infect transgenic mice expressing human mutant 102L PrP, human wild-type PrP or wild-type mice. We found that GSS-102L prions have properties distinct from other types of human prions by showing that they can only infect transgenic mice expressing human PrP carrying the same mutation. Mice expressing wild-type human PrP or wild-type mouse PrP were entirely resistant to infection with GSS-102L prions. We conclude that accurate modeling of inherited prion disease requires the expression of authentic mutant human PrP in transgenic models, as other approaches may generate results that do not mirror the human disease. Prion diseases are a closely related group of neurodegenerative conditions which affect both humans and animals ,2. They C) to alternative isoforms designated PrPSc sugtant PrP ,27. Herer clone) \u201350 minorutations \u201356.www.nc3rs.org.uk/ARRIVE/).Storage and biochemical analyses of post-mortem human brain samples and transmission studies to mice were performed with written informed consent from patients with capacity to give consent. Where patients were unable to give informed consent, assent was obtained from their relatives in accordance with UK legislation and Codes of Practice. Samples were stored and used in accordance with the Human Tissue Authority Codes of Practice and in line with the requirements of the Human Tissue Authority licence held by UCL Institute of Neurology. This study was performed with approval from the National Hospital for Neurology and Neurosurgery and the UCL Institute of Neurology Joint Research Ethics Committee \u2014REC references: 03/N036, 03/N038 and 03/N133. Work with mice was performed under approval and licence granted by the UK Home Office Act 1986; Project Licence number 70/6454) and conformed to University College London institutional and ARRIVE guidelines designated Tg(HuPrP102L 129M+/+Prnpo/o)-27 mice (102LL Tg27) [129M transgene array and murine PrP null alleles (Prnpo/o) designated Tg(HuPrP129M+/+Prnpo/o)-35 congenic (129MM Tg35c) were derived by subjecting previously described 129MM Tg35 mice [129V transgene array and murine PrP null alleles (Prnpo/o) designated Tg(HuPrP129V+/+Prnpo/o)-152 congenic (129VV Tg152c) were derived by subjecting previously described 129VV Tg152 mice [Transgenic mice homozygous for a human PrPLL Tg27) have beeg35 mice ,57 to co152 mice ,39,42 to2, and intra-cerebrally inoculated into the right parietal lobe with 30 \u03bcl of 1% (w/v) brain homogenate prepared in Dulbecco\u2019s phosphate buffered saline lacking Ca2+ or Mg2+ ions (D-PBS). All mice were thereafter examined daily for clinical signs of prion disease. Mice were killed if they exhibited any signs of distress or once a diagnosis of prion disease was established. At post-mortem brains from inoculated mice were removed, divided sagittally with half frozen and half fixed in 10% buffered formol saline.Strict bio-safety protocols were followed. Inocula were prepared, using disposable equipment for each inoculum, in a microbiological containment level 3 laboratory and inoculations performed within a class 1 microbiological safety cabinet. Ten mice per group from three transgenic lines, 102LL Tg27, 129MM Tg35c, 129VV Tg152c and FVB/N wild type mice were inoculated with a panel of prion isolates, all previously passaged in 102LL Tg27 transgenic mice and therefore adapted to human 102L PrP. The primary inocula comprised human brain homogenates from three IPD P102L patients, one sporadic CJD patient and three iatrogenic CJD patients. Diagnosis of all cases had been neuropathologically confirmed. The genotype of each mouse was confirmed by PCR of DNA prior to inclusion and all mice were uniquely identified by sub-cutaneous transponders. Disposable cages were used and all cage lids and water bottles were also uniquely identified by transponder and remained with each cage of mice throughout the incubation period. Care of the mice was according to institutional and ARRIVE guidelines. Mice were anaesthetised with a mixture of halothane and OPrnpo/o mice against \u03b1 or \u03b2 PrP as described elsewhere [1 with an epitope spanning residues 142\u2013153 of human PrP [2b with an epitope spanning residues 93\u2013105 of human PrP [Anti-PrP monoclonal antibodies ICSM 18 and ICSM 35 were supplied by D-Gen Ltd, London, UK. ICSM antibodies were raised in lsewhere . ICSM 18uman PrP . ICSM 35uman PrP ,59. ICSMuman PrP .Brain homogenates (10% (w/v)) were prepared in D-PBS and aliquots analysed in duplicate with or without proteinase K digestion by electrophoresis and immunoblotting as described previously ,61. Duplwww.ventana.com). Visualization was accomplished with diaminobenzidine staining. Bright field photographs were taken on an ImageView digital camera (www.soft-imaging.de) and composed with Adobe Photoshop.Fixed brain was immersed in 98% formic acid for 1 h and paraffin wax embedded. Serial sections of 4 \u03bcm nominal thickness were pre-treated with Tris-Citrate EDTA buffer for antigen retrieval . PrP dep"} +{"text": "Endothelial cell release of nitric oxide (NO) is one indicator of vascular health. Coronary arteries develop atherosclerotic disease, while left internal mammary arteries (IMA), which are frequently used as bypass conduits in patients with disease, do not. Endothelial cell function of vessels can be assessed by the response to isometric handgrip exercise (IHE); normal function is evidenced by an increase in cross sectional area(CSA), flow velocity(FV) and blood flow(BF), and an abnormal response by no increase in these variables. Recently, the combination of coronary MRI and isometric handgrip exercise (IHE) was shown to noninvasively quantify coronary endothelial function (CEF). We tested the hypotheses that: 1) IMA vasoreactivity to IHE is measurable using coronary 3T MRI 2) endothelial function of IMA differs from that of coronary arteries in patients with coronary atherosclerotic disease (CAD) whereas in healthy subjects endothelial function of the two vascular beds is similar and 3) the IMA response to IHE is primarily mediated by NO, thus reflecting endothelial function.We studied 21 CAD patients (60\u00b12years) and 27 healthy subjects (46\u00b14years) on a commercial 3T MRI scanner using a 32-element cardiac coil. Anatomical and velocity-encoded spiral cine sequences were obtained of both the right coronary artery (RCA) and an IMA in axial cross-section in the same cine sequence Fig at rest During IHE in healthy subjects, mean CSA, FV and BF for both RCA and IMA increased significantly from baseline Fig . As expeBoth coronary and systemic endothelial function can now be measured in a single MRI acquisition. The IMA response to IHE is significantly attenuated by L-NMMA indicating the IMA vasoreactive response to IHE primarily reflects NO-dependent endothelial function. Coronary and systemic vascular beds display heterogeneous responses, which promise unique pathophysiologic insights in atherosclerotic and atherosclerosis-free vascular beds.NIH/NHLBI grants R01HL084186, AHA SDG 5200004, AHA 12PRE11510006."} +{"text": "There are errors in the published article. The isolate # 3172 ABBSB1189-1 (genome 14) was named as Enteritidis by mistake. The correct isolate should be Typhimurium.Serotyping was incorrect for the isolate # 3193 ABBSB1050-2 (genome 13). It was incorrectly labeled as Enteritidis. The correct label should be Thompson.These errors affect Figs S3 FigEscherichia coli clade is collapsed into a single branch. Numbers at internal nodes correspond to bootstrap support values. *** indicates the 25 newly sequenced Salmonella genomes of this study.The (TIFF)Click here for additional data file.S4 FigAnother 111 ubiquitous proteins are not shown. Labels and colors are consistent with (TIFF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file."} +{"text": "During the process of T cell priming to peptide antigens, the T cell receptor (TCR) undergoes gene rearrangements that result in a peptide-binding region which can act as a unique molecular fingerprint for similar T cell clonotypes. The purpose of this study was to assess high-throughput TCR DNA sequencing to analyze qualitative and quantitative measures of human T cell clonotype expansions both in vitro and in vivo.Human lymph nodes derived from patients with melanoma were culture-activated using either anti-CD3/CD28 beads and IL-2 100 IU/ml alone or in combination with anti-VEGF neutralizing antibody, for 14 days. The two resultant MDLN cultures were compared for overall T cell expansion, cell surface phenotype, intracellular cytokine staining and high-throughput TCR DNA sequencing . In addition, both TDLN cell cultures were injected intravenously into SCID mice bearing A375 melanoma xenografts and bone marrow was harvested to assess for persistence of transferred human melanoma MDLN cells.Overall T cell expansion and cell surface phenotype of MDLN cell cultures was similar. MDLN cells cultured in the presence of anti-VEGF neutralizing antibody demonstrated higher baseline levels of intracellular interferon-\u03b3. TCR sequencing analysis comparing day 0 versus day 14 cultured MDLN cells demonstrated approximately 10% shared T cell clonotypes following culture activation with anti-CD3/CD28 beads and IL-2. As a point of reference, the proportion of shared T cell clonotypes between two different patient MDLN cells was approximately 0.6%. Addition of anti-VEGF antibody during culture of MDLN cells resulted in common T cell clonotypes of approximately 1% (day 0 versus day 14), confirming the expansion of molecularly distinct T cell clonotypes. Human MDLN cells infused into SCID mice bearing A375 melanoma xenografts demonstrated persistence of T cell clonotypes in the bone marrow several weeks after infusion. It is notable that none of the low frequency T cell clonotype expansion differences between samples could be readily identified using VDJ gene usage or CDR3 length analysis.This study demonstrates that CDR3 sequence data that is quantitative and recognizes low frequency T cell clonotypes is more sensitive than V gene usage and CDR3 length analysis. In addition, CDR3 sequencing may provide utility in the qualitative assessment of T cells both in vitro and in vivo."} +{"text": "Functional ankle instability (FAI) has been reported to be associated with sensorimotor deficits which could result in impaired balance and alte20 athletes (age: 26.55\u00b15.35years) with clinically diagnosed FAI were recruited. Static balance in double limb stance was assessed using Kistler force plates during four shod conditions: 1) flat EVA base insole 2) Textured flat EVA insole 3) Lateral heel and sole wedge 4) Textured lateral heel and sole wedge . Texture was a semirigid rubber with semi-circular mounds with center to center distances of 4 mm. The center-of-pressure excursion and mean velocity in anterior-posterior and medial-lateral directions and area of 95% confidence circle were derived as measures of standing balance. The results were statistically analyzed using the nonparametric Fridman test followed by Wilcoxon Signed Rank.Statistically significant differences were observed only for the textured flat EVA insole. The mean COP velocity was reduced compared to the lateral wedge condition (p <0.05) and the 95% confidence circle area decreased significantly compared with all other insoles conditions Table .There weTexture appears to have some impact on standing balance but only on a flat insole. The lateral wedge had no effect on standing balance.Nester declares a personal commercial interest in the insoles tested in this study."} +{"text": "However, her seizures were becoming more frequent despite being on Carbamazepine and Phenobarbital at maximal tolerable dosages. She had no recent history of head trauma neither any other medical illness. Her physical examination was unremarkable. Her full blood count, liver and kidney function tests were within normal limits. A Brain CT scan was ordered which revealed multiple calcified and vesicular cysts within the Brain parenchyma; features consistent with Neurocysticercosis. The stool examination was negative for Taenia species ova. She was treated with Albendazole 400mg tid for 1 month and subsequent CT scans at 4 weekly intervals showed marked reduction in the number of active (vesicular cysts). Brain CT scans showing multiple vesicular cysts before treatment with Albendazole (A and B) and after treatment (C and D)"} +{"text": "MFN2, GJB1, BSCL2, and SETX, are previously reported mutations and considered to be pathogenic in these families. Twelve novel variants (11%) in the genes NEFL, TRPV4, KIF1B, BICD2, and SETX are implicated in the disease but require further evidence of pathogenicity. The remaining 20 variants were confirmed as polymorphisms (not causing the disease) and are detailed here to help interpret sequence variants identified in other family studies. Validation using segregation, normal controls, and bioinformatics tools was valuable as supporting evidence for sequence variants implicated in disease. In addition, we identified one SETX sequence variant (c.7640T>C), previously reported as a putative mutation, which we have confirmed as a nonpathogenic rare polymorphism. This study highlights the advantage of using WES for genetic diagnosis in highly heterogeneous diseases such as IPNs and has been particularly powerful in this cohort where genetic diagnosis could not be achieved due to phenotype and mode of inheritance not being previously obvious. However, first tier testing for common genes in clinically well-defined cases remains important and will account for most positive results.Inherited peripheral neuropathies (IPNs) are a group of related diseases primarily affecting the peripheral motor and sensory neurons. They include the hereditary sensory neuropathies (HSN), hereditary motor neuropathies (HMN), and Charcot-Marie-Tooth disease (CMT). Using whole-exome sequencing (WES) to achieve a genetic diagnosis is particularly suited to IPNs, where over 80 genes are involved with weak genotype\u2013phenotype correlations beyond the most common genes. We performed WES for 110 index patients with IPN where the genetic cause was undetermined after previous screening for mutations in common genes selected by phenotype and mode of inheritance. We identified 41 missense sequence variants in the known IPN genes in our cohort of 110 index patients. Nine variants (8%), identified in the genes Inherited peripheral neuropathies (IPNs) are a group of related diseases primarily affecting the peripheral motor and sensory neurons. They include the hereditary sensory neuropathies (HSN), hereditary motor neuropathies (HMN), and Charcot-Marie-Tooth disease (CMT). There is significant genetic and clinical heterogeneity in IPNs. To date, mutations have been described in over 80 genes that cause CMT and related disorders has been shown to be an efficient tool for mutation screening CMT disease .n\u00a0=\u00a076), HMN (n\u00a0=\u00a014), HMN with pyramidal tract signs , and HSN (n\u00a0=\u00a01). Families with CMT were further classified into axonal CMT , demyelinating CMT , CMT with pyramidal signs (n\u00a0=\u00a02), and CMTX (n\u00a0=\u00a027). Families with no male-to-male transmission and with milder phenotype in females were classified as X linked (CMTX). However, due to the small size of these pedigrees, autosomal dominant inheritance with reduced disease penetrance could not be excluded.The 110 index patients were from families with multiple affected individuals except for two sporadic cases which could represent recessive inheritance or sporadic cases. Patient selection was from families with unknown genetic etiology after genetic testing for common IPN genes as described below. Index patients had various IPN phenotypes; CMT (PMP22 (MIM 601097) and Sanger sequencing for point mutations in PMP22, MPZ (MIM 159440), GJB1 (MIM 304040), MFN2 (MIM 608507), SPTLC1 (MIM 605712) and SPTLC2 (MIM 605713). WES of DNA from index patients was outsourced to Axeq Technologies (South Korea). In brief, sequencing libraries were prepared using the Illumina TrueSeq kit and sequenced as 100\u00a0bp paired-end reads on the HiSeq 2000 Sequencer . Sequence reads were aligned to the Human Genome Feb\u00a02009 (GRCh37/hg19) assembly using BWA software (Li and Durbin http://picard.sourceforge.net/). Sequence variants (single nucleotide variants and indels) were called using SAMTOOLS , AIFM1 (MIM 300169), ARHGEF10 (MIM 608136), ATL1 (MIM 606439), ATP7A (MIM 300011), BSCL2 (MIM 606158), CCT5 (MIM 610150), COX6A1 (MIM 602072), DARS (MIM 603084), DCTN1 (MIM 601143), DHTKD1 (MIM 614984), DNAJB2 (MIM 604139), DNM2 (MIM 602378), DNMT1 (MIM 126375), DYNC1H1 (MIM 600112), EGR2 (MIM 129010), FAM134B (MIM 613114), FBLN5 (MIM 604580), FGD4 (MIM 611104), FIG 4 (MIM 609390), GARS (MIM 600287), GDAP1 (MIM 606598), GJB1, GNB4 (MIM 610863), HINT1 (MIM 601314), HK1 (MIM 142600), HSPB1 (MIM 602195), HSPB3 (MIM 604624), HSPB8 (MIM 608014), IFRD1 (MIM 603502), IGHMBP2 (MIM 600502), IKBKAP (MIM 603722), INF2 (MIM 610982), KARS (MIM 601421), KIF1A (MIM 601255), KIF1B (MIM 605995), LITAF (MIM 603795), LMNA (MIM 150330), LRSAM1 (MIM 610933), MED25 (MIM 610197), MFN2, MPZ, MTMR2 (MIM 603557), NDRG1 (MIM 605262), NEFL (MIM 162280), NGFB (MIM 162030), NTRK1 (MIM 191315), PDK3 (MIM 300906), PLEKHG5 (MIM 611101), PMP22, PRPS1 (MIM 311850), PRX (MIM 605725), RAB7A (MIM 602298), REEP1 (MIM 609139), SBF1 (MTMR5) (MIM 603560), SBF2 (MTMR13) (MIM 607697), SETX (MIM 608465), SH3TC2 (MIM 608206), SLC12A6 (MIM 604878), SLC5A7 (MIM 608761), SOX10 (MIM 602229), SPTLC1, SPTLC2, SURF1 (MIM 185620), TDP1 (MIM 607198), TFG (MIM 602498), TRIM2 (MIM 614141), TRPV4 (MIM 605427), VAPB (MIM 605704), VCP (MIM 601023), WNK1 (MIM 605232), and YARS (MIM 603623). The sequence variants that were considered for further study included; nonsynonymous, frameshift, splicing, small indels, upstream promoter, and regulatory and UTR variants. The annotation and nomenclature of sequence variants were checked using the web based software Mutalyzer 2.0.beta-29 .Prior to this study, patients were screened for mutations in common IPN genes based on clinical presentation. These genes included; MLPA duplication analysis of R (R Core Team Novel variants were assessed to predict if they were likely to cause disease. DNA and protein sequences were obtained from the UCSC Genome Browser (GRCh37/hg19) . Therefore, our WES data have >90% sensitivity to detect heterozygous variants within the exome target regions .WES was performed on 110 index patients from unrelated peripheral neuropathy families. Two samples with previously identified point mutations in WES initially identified 62 sequence variants in 20 known IPN genes. Twenty variants were excluded as false positives by Sanger sequencing. Of the remaining 41 sequence variants, 21 were excluded as rare or novel polymorphisms by segregation analysis or from previously published evidence . The proIn 21 of the 110 index patients, we identified a sequence variant that caused a missense mutation in a known CMT or HMN disease gene Table. The preMFN2 mutations, c.775C>T (p.R259C) and c.1090C>T (p.R364W) and no retrospective testing was requested in these cases.Two previously reported GJB1; c.77C>T (p.S26L) in which a carrier female with mild peripheral neuropathy had two sons with a CMT1 phenotype. Family F, with the p.R264C mutation had only two affected females (mother and daughter), with no at-risk males. Both women had a juvenile onset axonal HMN phenotype presenting as weakness of the feet and ankles, with later progression to include pes cavus, hammertoes, and muscle wasting of the feet. There was no sensory involvement and normal nerve conduction velocities. As there were no affected males in this pedigree, it is unknown if males would present with a more severe phenotype than the females in the family. This mutation was previously reported to cause an axonal CMT phenotype a method that can lead to false negatives if the mutations are located between melt domains or if signals are quenched by secondary SNPs was identified in index individuals from two unrelated families mutation in SETX was identified in an index individual diagnosed with HMNP affects the same amino acid as a previously reported mutation p.L268P (rs62636502) was identified in a family with axonal CMT (Family N). The proband had muscle weakness in the hands and distal legs, with sensory loss and neuropathic pain in the feet. While predominantly a peripheral neuropathy the patient showed clear upper motor neuron involvement with facial weakness. The disease showed autosomal dominant inheritance with a strong family history of neuromuscular disease. The c.794A>G sequence variant was confirmed in the proband and three additional affected individuals and was not present in one unaffected individual and one married-in spouse. While this nucleotide is conserved, bioinformatics analyses were contradictory making it difficult to determine the likely pathogenicity status of this variant was identified in a patient with axonal CMT with asymmetrical gait (Family O). The inheritance pattern of the disease in this family is unclear and no other DNA samples were available for testing. Bioinformatics analysis suggested the variant is likely to cause disease (Table36P was iThe novel sequence variant c.1319C>T (p.P440L) was identified in an autosomal dominant family with axonal CMT (Family P). Affected individuals had upper and lower limb muscle weakness and wasting, slight peroneal weakness , brisk reflexes (upper and lower), and a possible extensor plantar response. Some individuals also had reduced sensory action potentials, whereas others had no sensory involvement. Segregation of p.P440L NEFL in the family is unclear. The variant was present in all tested affected individuals and absent in one unaffected (at-risk) individual. However, it was also present for an at-risk family member of unknown clinical phenotype. Some late onset cases have been described for the NEFL mutations was identified in the index individual from Family Q BICD2 variant is likely to be pathogenic as the variants are not well conserved and both prediction programs suggested the variant changes are benign variant of the cohort, with an additional 11% (12 of 110) with novel sequence variants that are yet to be proven pathogenic. Prior genetic testing has identified most patients that have mutations in common genes. The overall contribution of the known disease genes in CMT previously published is 67%, with over 90% of this genetic diagnosis described by four common genes . On review, both MFN2 mutations were identified in families last clinically examined prior to MFN2 testing becoming available, and this study was the first genetic review of each family. Of the families with GJB1 mutations, none had male-to-male inheritance. In three families, X-linked inheritance was not previously recognized due to no affected males with sons and a limited number of males and females for phenotype variability to effectively assign this mode of inheritance. The remaining family with GJB1 was suspected of being X linked and previously tested for GJB1 (year 1997), with a negative result which is likely due to reduced sensitivity of mutation scanning methods used at that time. The implementation of WES in a diagnostic setting is unlikely to increase the overall genetic diagnostic success rate (based on standard MLPA/Sanger sequencing methodologies), but in future may lead to reduced time and costs for samples prescreened for the CMT1A duplication which was previously reported as a putative cause of a sporadic ataxia-tremor and motor neuron disease phenotype in a single individual . Additionally, we correctly identified the mutation for two positive control samples as well as AARS SNPs (McLaughlin et\u00a0al. MT-ATP6 (Pitceathly et\u00a0al. We used the mean depth of coverage as a measure of the sensitivity of WES to identify mutations. We estimated the sensitivity to detect mutations in IPN genes in this cohort is in the range of 90\u201398% (Choi et\u00a0al. In conclusion, we have successfully applied WES to our cohort of IPN cases and identified pathogenic sequence variants in 8% of cases and novel sequence variants that may cause disease in an additional 11% of our cohort. This study has highlighted the advantage of using WES for genetic diagnosis in highly heterogeneous diseases such as the IPNs and has been especially powerful when inheritance is unclear. However, first tier testing for common genes in clinically well-defined cases remains important and will account for most positive results. Research into the genetic cause of the remaining cases will identify new disease genes furthering our understanding of the biology underlying IPNs."} +{"text": "Cancer immunotherapy has generated significant response rates and prolonged survival, particularly in metastatic melanoma, but carries the risk of immune-related adverse events (irAEs) [1]. Treatment of elderly patients with checkpoint inhibition presents a unique challenge as nearly half of all malignancies are diagnosed in patients >65 [2], and clinical indications for immunotherapy continue to increase. Side effects may be more challenging in older patients given the association of age with comorbidties and reduced functional reserve [3]; additionally \u201cimmunosenescence,\u201d age-related impairment of adaptive and innate immunity, could impair effective checkpoint inhibition [4].Retrospective analysis of irAEs and survival outcomes in melanoma patients < 65 compared to those >65 treated with nivolumab.Data pooled from 148 patients treated with nivolumab plus peptide vaccine or nivolumab alone every two weeks for at least 12 weeks. Frequency, grade, and characteristics of irAEs were analyzed among patients >65 and < 65 years of age. A 12-week landmark survival analysis was then assessed for each group.Of 148 patients, 52 (35%) were age >65. Most common irAEs among pts < 65 included diarrhea/colitis (30.2%), rash (38.5%), and vitiligo (10.4%). These were also the most common irAEs in pts > 65 . No statistically significant difference in irAE incidence was seen between groups and there was no statistically significant OS difference between patients >65 and < 65 (p=0.115). A statistically significant OS benefit was seen in patients < 65 and >65 experiencing any grade of irAE .Nivolumab may be an effective therapeutic option for patients 65 and older, as the irAE profile seen below and above this age cutoff was similar, and showed no statistically significant difference in incidence. Additionally the presence of irAEs in patients >65 was still associated with survival benefit, thus demonstrating such patients may still respond appropriately to checkpoint inhibition. These encouraging data should be validated in larger patient cohorts, with specifically targeted age ranges to further investigate tolerance and response."} +{"text": "Incidence of anomalous coronary anatomy (ACA) in caucasian and middle eastern patients with ventricular septal defect (VSD) and pulmonary stenosis (PS) is between 2-9%. Prevalence of ACA in South Asian (SA) patients is unknown. Patients with VSD and PS may have Tetrology of Fallot (TOF) or Double Outlet Right ventricle (DORV) morphology. This study looks at the prevalence of ACA in SA patients with TOF/DORV. Pre-operative assessment of ACA is of paramount importance in these patients as it alters surgical procedure.Retrospective analysis of all Multi Detector Computed Tomography (MDCT) and Cardiac Magnetic Resonance (CMR) examinations of SA patients with TOF/DORV was performed in our institute over 4 years (June 2011 to June 2014). Coronary anomalies were classified into Variant Coronary Anatomy (VCA) and ACA. VCA included coronaries that coursed close to the right ventricular outflow tract (RVOT) while ACA included anomalous origin or branching of coronaries.A total of 1011 scans were performed during this period. Of these, 95 were excluded due to suboptimal visualisation of the coronary arteries. Coronary anomalies were seen in 146 (16%) patients. Most common abnormality was VCA, seen in 78 (8.5%) patients with large conus or acute marginal or septal branch crossing the RVOT. ACA was seen in 68 patients (7.4%), with left anterior descending (LAD)/ Left main stem (LMS) arising from right coronary sinus in 35 patients (4%) and right coronary artery (RCA) arising from LAD/LMS in 16 patients (1.7%). OTher anomalies included left coronary artery origin from pulmonary artery (ALCAPA), coronary cameral fistula, single coronary arteries, dual LAD and conus branch from coronary sinus.Coronary artery anomalies are common in SA patients with VSD and PS. Optimal imaging is therefore essential to establish coronary anatomy prior to surgery."} +{"text": "We sought to optimize the Nasal Allergen Challenge (NAC) model to ensure reliability and repeatability of results by modifying the qualifying criteria and allergen concentration during the challenge.20 Allergic Rhinitis (AR) participants underwent NAC to determine the concentration at which a Total Nasal Symptom Score (TNSS) of 10/12 OR a Peak Nasal Inspiratory Flow (PNIF) reduction of 50 % was achieved. 4-fold increases in allergen concentration were administered every 15 minutes until qualification criteria were met. The Qualifying Allergen Concentration (QAC) reached was used as a single challenge dose at the subsequent NAC visit. 10 additional ragweed allergic and 4 non-allergic participants were qualified at a TNSS of 8/12 AND a PNIF reduction of 50%. Cumulative Allergen Concentration (CAC) of all incremental doses was used during the subsequent NAC visit. Participants recorded TNSS and PNIF at baseline, 15 minutes, 30 minutes, 1 hour and hourly afterwards up to 12 hours post-challenge during the NAC visit.QAC study participants qualifying only based on PNIF reduction had significantly lower TNSS scores than those qualifying on TNSS only or TNSS+PNIF (p<0.01). Participants in both studies\u2019 NAC visit reached peak TNSS at 15 minutes post-challenge followed by a gradual symptom decline, while the \u201cPNIF only\u201d group had significantly lower TNSS compared to others. All 3 groups experienced a decline in peak TNSS following NAC compared to screening, although groups qualifying on TNSS and TNSS+PNIF maintained their PNIF scores.The NAC model is well-suited to study AR symptoms. TNSS and PNIF are complementary and must be integrated in the qualifying criteria. Further protocol modifications, such as with multiple allergen challenges during the NAC visit, may produce even more repeatable results. Through optimizing the NAC protocol, the model achieves reproducible results and becomes more reliable; suitable for testing new medications in clinical trials."} +{"text": "Diabetiear 2030 .The aim of this study is to develop a methodology for automatic detection of patients at risk for PU based on 3 dimensional (3D) multisegment foot biomechanics through cluster analysis.For this purpose 44 subjects, 20 with (PN) and 24 without PN (noPN) were enrolled. Simultaneous kinematic, kinetic and plantar pressure (PP) data were acquired during gait with a BTS motion capture system synchronized with 2 Bertec force plate (FP4060-10) and 2 Winpod pressure plate as in , electroA hierarchical cluster (HC) technique was adopted using TiResults of HC analysis ."} +{"text": "The concept of external hydrocephalus refers to situations of CSF flow impairment within subarachnoid spaces (SAS). Classically described in infant and children, literature offers few data on adult.We retrospectively analysed adult patients admitted in four French intensive care units, between November 2010 and December 2014, for severe traumatic brain injury (TBI) or subarachnoid hemorrhage (SAH). We undertook clinical and radiological findings of patients presenting intracranial hypertension (ICHT) presumably in relation with external hydrocephalus , treated with cerebrospinal fluid (CSF) external lumbar drainage (ELD).33 patients admitted for TBI (n=22), SAH (n=8) or other brain insults (n=3) with a mean initial Glasgow score of 8 (+/- 4) were included. 25 (75.8%) patients did not receive former external ventricular drainage. In all cases, ELD was dramatically effective to lower intracranial pressure (25.2 mmHg [+/-9.1] before EDL vs 7.4 mmHg [+/-6.0] after EDL). No mydriasis or intracranial bleeding occurred. One patient (3%) developed an ELD infection. Patients were discharged from ICU with a mean modified Rankin Score of 4[+/-1].Often described as a passive process (e.g. hygroma), CSF accumulation around the brain after acute cerebral insults in adults can be approached as an active process of external hydrocephalus. This diagnosis remains often subtle, but should systematically be evoked when CT scan show paradoxical enlargement of subarachnoid spaces in a context of ICHT. Our data tend to confirm that in these specific situations, ELD should be considered as a safe, effective and minimal invasive option."} +{"text": "Advances in the field of plant regeneration show that the first steps of de novo organogenesis through in vitro culture in hormone containing media require root post-embryonic developmental programs as well as regulators of auxin and cytokinin signaling pathways. We review how hormonal regulation is delivered during lateral root initiation and callus formation. Implications in reprograming, cell fate and pluripotency acquisition are discussed. Finally, we analyze the function of cell cycle regulators and connections with epigenetic regulation. Future work dissecting plant organogenesis driven by both endogenous and exogenous cues may reveal new paradigms of common regulation.Plants have extraordinary developmental plasticity as they continuously form organs during post-embryonic development. In addition they may regenerate organs upon Cyperus papyrus or Rubus fruticosus). These regeneration capabilities of plants have been exploited in agriculture for propagation purposes of selected varieties, virus sanitization and development of biotechnological tools revealed a series of common mechanisms and regulators. In this review, we summarize recent advances in the field and discuss parallelisms between both processes.During post-embryonic developmental programs lateral meristems are formed ell fate . This naell fate . CompariArabidopsis thaliana, development of new aerial organs is initiated by the plant hormone auxin meristems are initiated in the differentiation zone of the root from pairs of founder cells that derive from the pericycle . The term callus had been previously used to designate outgrowth of cells associated with callose accumulation and wounding . Different concentrations of these hormones regulate the balance between cell proliferation and differentiation and callus formation is regulated by the hormones auxin, cytokinin and their downstream signaling pathways callus at different time points during CIM incubation (As auxin and cytokinin downstream pathways operate both during post-embryonic and cubation and (2) cubation , shows ccubation . 847 genARABIDOPSIS 1 (TAA1) acts in the indole-3-pyruvic acid branch of the auxin biosynthetic pathway. Loss of function mutants of TAA1 (tir2) have impaired LR formation that can be rescue by treatment with auxin and the auxin biosynthetic intermediate like indole-3-pyruvic acid shows a strong phenotype with no LR formation. This mutation blocks asymmetric cell divisions of LR founder cells during LR initiation proteins, which function transcriptionally as activators or repressors , and AUXitiation . A gain-itiation . Furtherenotypes . Interesormation . AlthougLBD/ASL genes are rapidly activated by CIM. Furthermore, ectopic expression of these LBDs/ASLs can induce callus formation without plant hormones treatment; whereas T-DNA insertion mutants (lbd16-2 and lbd18-1), inhibit callus formation. These four LBD/ASL factors are downstream of ARF7 and ARF19 (AUXIN RESPONSE FACTORs regulates LATERAL ORGAN BOUNDARIES-DOMAIN/ASYMMETRIC LEAVES2-LIKE-(LBD/ASL) transcription factors during LR organogenesis. It has been described that ARFs primarily regulate LR initiation via direct activation of itiation . A recennd ARF19 . This innd ARF19 , it appeCytokinin is an important component of CIM. de novo organogenesis from callus or explants. High concentration of cytokinin reduces root formation to promote shoot differentiation. Cytokinin has been described to inhibit root identity genes when callus are transferred to shoot induced medium . These proteins can be classified in two groups, type A and type B. Type A are rapidly upregulated by exogenous cytokinin and repress cytokinin signaling. In Arabidopsis, there are 10 members of the type A group has more free auxin and shows excessive proliferation of pericycle cells in mature parts of the root. These dividing pericycle cells do not differentiate into LRs promotes transcriptional activation of the cell cycle . However, there is no evidence that E2Fa overexpression induces callus formation even when co-expressed with its dimerizing partner DPA CKS2 or the cell division protein APC6 (cyca2s) display reduced LR density and deviations in LR primordium patterning . Overexpression of ESR2 induces callus without hormonal treatment and shows elevated cytokinin response -RELATED PROTEIN (ICK/KRP) are down regulated after auxin treatment, being the transcriptional adaptor protein PROPORZ1 (PRZ1) responsible of this repression or CDKs is not sufficient to induce callus or LR development . Howeverg medium . Callus Callus formation requires dramatic changes in both cell identities and cell growth patterns. These changes have been shown to be accompanied by activation or repression of numerous genes across the genome . It seemArabidopsis proteins CURLY LEAF (CLF), SWINGER (SWN), VERNALIZATION (VRN2) and EMBRYONIC FLOWER2 (EMF2) participate as core components of PRC2 proteins act in an evolutionarily conserved epigenetic pathway that regulates chromatin structure. PcG proteins repress many developmentally important genes through modification of histones. PcG proteins can form at least two multiprotein complexes: the Polycomb repressive complexes 1 and 2 (PRC1 and PRC2). In plants, the major function of PRC2 is to trimethylate lysine 27 on histone H3 (H3K27me3); while PRC1 recognizes the H3K27me3 marker and mono-ubiquitinates histone H2Aub . Both ep of PRC2 . Double of PRC2 . Similar of PRC2 . Double ormation . Thus itormation , shoot fArabidopsis AtBM1A and AtBM1B genes are the homologs of mammalian PRC1 gene , LEAFYCOTYLEDON2 (LEC2), AGAMUS-LIKE 15 (AGL15), or BABY BOOM (BBM). As over expression of embryonic fate regulators results in formation of ectopic embryos also plays a significant role in transcriptional repression of cell identity genes and pkl mutants spontaneously develop callus after germination mutant turns out to be another mutant allele of PKL. In addition, treatments with TSA partially suppress the phenotype of slr1 at the molecular level. Example of this is the LR inducible system, which has been useful to find novel regulators of LR formation but also of stem cell niche function . FinallyThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Neuromyelitis optica (NMO) is a severely disabling inflammatory disorder of the central nervous system and is often misdiagnosed as multiple sclerosis (MS). There is increasing evidence that treatment options shown to be beneficial in MS, including interferon-\u03b2 (IFN-\u03b2), are detrimental in NMO.We here report the first Caucasian patient with aquaporin 4 (AQP4) antibody (NMO-IgG)-seropositive NMO presenting with a tumefactive brain lesion on treatment with IFN-\u03b2. Disease started with relapsing optic neuritis and an episode of longitudinally extensive transverse myelitis (LETM) in the absence of any brain MRI lesions or cerebrospinal fluid-restricted oligoclonal bands. After initial misdiagnosis of multiple sclerosis (MS) the patient received subcutaneous IFN-\u03b21b and, subsequently, subcutaneous IFN-\u03b21a therapy for several years. Under this treatment, the patient showed persisting relapse activity and finally presented with a severe episode of subacute aphasia and right-sided hemiparesis due to a large T2 hyperintensive tumefactive lesion of the left brain hemisphere and a smaller T2 lesion on the right side. Despite rituximab therapy two further LETM episodes occurred, resulting in severe neurological deficits. Therapeutic blockade of the interleukin (IL)-6 signalling pathway by tocilizumab was initiated, followed by clinical and radiological stabilization.Our case (i) illustrates the relevance of correctly distinguishing NMO and MS since these disorders differ markedly in their responsiveness to immunomodulatory and -suppressive therapies; (ii) confirms and extends a previous report describing the development of tumefactive brain lesions under IFN-\u03b2 therapy in two Asian NMO patients; and (iii) suggests tocilizumab as a promising therapeutic alternative in highly active NMO disease courses. Interferon-\u03b2 (IFN-\u03b2) is one of the established first-line therapies for patients with relapsing-remitting multiple sclerosis (MS). While the efficacy of IFN-\u03b2 in MS is widely accepted, the possible benefit in non-classical MS variants has been disputed. This is the case for neuromyelitis optica (NMO/Devic\u2019s syndrome), considered an MS subtype until the discovery of the anti-aquaporin 4 (AQP4) antibodies (AQP4-Ab) ,2. In boA 47-year old Caucasian female patient had been diagnosed with MS in 1995 in an external hospital after four episodes of optic neuritis (ON) since 1989 and an episode of longitudinally extensive transverse myelitis (LETM) in 1995 in May 2013 and transferred our patient to a rehabilitation centre. However, further severe LETM attacks occurred shortly thereafter in June and August 2013. The patient was re-admitted to our department in September 2013 and showed persistently increased AQP4-Ab titres . Absolute B cell counts were normal (68/\u03bcl) while relative proportion was decreased . This relapse was associated with a urinary tract infection levels) which we successfully treated according to local guidelines. In light of the patient\u2019s severe clinical deficits (EDSS of 9.0) we applied another iv steroid pulse and decided to switch to tocilizumab, a humanized monoclonal anti-IL-6-receptor antibody. Monthly infusions of tocilizumab with 8 mg/kg bodyweight were followed by clinical stabilization as no further relapses occurred for the following 12 months (see clinical and MRI follow-up after withdrawal of IFN-\u03b2 in Figure\u00a0To our knowledge, we here present the first Caucasian NMO patient who developed a tumefactive brain lesion on long-term IFN-\u03b2 treatment. Our case confirms and extends similar findings in two Asian NMOSD patients recently reported by Shimizu and colleagues . ObviousOf note, administration of a single intravenous infusion of 1000 mg rituximab was followed by further relapses just one and three months later. While CD19 counts were not determined directly after rituximab application, an investigation four months after rituximab dosing showed normal absolute B cell counts and persistently increased AQP4-Ab levels. We may speculate that our patient belongs to individuals characterized by an early repopulation of the B cell compartment following rituximab therapy .ex vivo experimental model of NMO spinal cord slice cultures exposed to NMO-IgG and complement showed a marked loss of AQP4 and myelin which was enhanced by adding IL-6 [+ CD27+ CD38+ CD180+ B cells were found to produce AQP4 antibodies and showed enhanced survival as well as AQP4 antibody secretion in the presence of IL-6, whereas blockage of the IL-6 receptor signalling by an anti-IL-6R antibody shortened their survival in vitro [Our case adds preliminary evidence in favour of a possible beneficial effect of tocilizumab in NMO, even following treatment with rituximab -15. Howeing IL-6 . Moreovein vitro .Our report illustrates the importance of correctly diagnosing NMO and MS to avoid mistreatment with potentially severe or even fatal consequences. Two recent studies have revealed that up to 30% of patients with NMO were falsely diagnosed with MS ,22 and, Our case supports the conclusion that the development of tumefactive brain lesions under IFN-\u03b2 therapy for suspected MS should prompt considering NMOSD as the underlying disease, obviously not only in Asian but also in Caucasian patients. Moreover, our report underlines \u2013 in accordance with previous reports and recommendations \u2013 the neWritten informed consent was obtained from the patient for publication of this case report and any accompanying images. A copy of the written consent is available for review by the Editor of this journal."} +{"text": "Accumulation of diffuse interstitial myocardial fibrosis in severe aortic stenosis relates to left ventricular (LV) functional impairment and determines the postoperative outcome in these patients. T1 mapping has been proposed for non-invasive quantification of diffuse interstitial fibrosis. We examined associations between T1 values and collagen volume fraction from endomyocardial biopsies from patients with severe aortic stenosis. We compared whether these relationships differ with respect to the post-processing approach.Ten patients with isolated severe aortic stenosis and eligible for aortic valve replacement surgery underwent clinical cardiovascular magnetic resonance study at 3 clinical scanner. A mid- chamber myocardial biopsy was obtained from the left ventricular (LV) septum at the time of surgery and stained for collagen volume fraction (CVF) using Mason Trichrome technique and ImageJ analysis.T1 mapping was performed using MOLLI (3(3)3(3)5)) sequence prior to and 15 minutes after IV administration of 0.2 mmol/kg of gadobutrol. T1 measurements were performed using both a conservative septal ROI and the entire coverage of midventricular short-axis (SAX) slice. Areas of overt LGE were excluded from the analysis. T1 values were compared to fourteen age-matched healthy control subjects and correlated with CVF.Compared to controls, there were no significant differences in cardiac volumes and global systolic function. Patients had raised LV mass and wall thickness p<0.01), and mean native T1 values comprehensive Biomedical Research Centre award to Guy's & St Thomas' NHS Foundation Trust in partnership with King's College London and King's College Hospital National Health Service Foundation Trust, and British Heart Foundation Research Centre of Excellence Award. King's Health Partners via MRC Confidence in Concept Grant."} +{"text": "An abundance of research is available concerning exercise and caffeine with men, but among women resistance trainers, it is scarce. We tested a highly caffeinated coffee product, Via\u00ae instant coffee (VIA) vs. its decaffeinated version (DCF) on men and women. We hypothesized that VIA consumption would not benefit men greater than women (p>0.05) when comparing muscle explosiveness among resistance-trained (6.4\u00b13.7 y) college students .After 24 hours of dietary control and caffeine abstinence, fasted subjects volunteers to perform three separate repetitions of strict Smith bench press (30% 1RM) under two conditions , with conditions separated by 48-72 hours. The peak force (FOR), peak power (POW), peak velocity (VEL), and maximum rate of force development (RFD) of the VIA trial were compared to DCF. FOR, POW, VEL, and RFD were measured via Ballistic Measurement System (BMS) linear displacement .The interaction of coffee and gender were analyzed in both absolute and relative terms via 2x2 ANOVA with repeated measures (covariance as necessary) and Tukey HSD post hoc . Consent to publish the results was obtained from all participants.VIA enhanced absolute POW for men and for women ; absolute VEL for men and for women ; and RFD for men but not for women . Further, absolute enhancement was greater for men in POW (p=0.037) and in RFD (p=0.028). No gender differences persisted when enhancement was expressed as percent increases over baseline (DCF) or after statistically adjusting for body mass.These data support the hypothesis that men and women benefit similarly (7-10%) after ingesting two servings of Via instant coffee approximately one hour before Smith bench press exercise."} +{"text": "In contrast to Western medicine, which typically prescribes one medicine to treat a specific disease, traditional East Asian medicine uses any one of a large number of different prescriptions , according to the patient's characteristics. Although this can be considered an advantage, the lack of a universal prescription for a specific disease is considered a drawback of traditional East Asian medicine. The establishment of universally applicable prescriptions for specific diseases is therefore required. As a basic first step in this process, this study aimed to select prescriptions used in the treatment of stroke and, through the analysis of medicinal herb combination frequencies, select a high-frequency medicinal herb combination group for further experimental and clinical research. As a result, we selected some candidates of a medicinal herb combination and 13 candidates of a medicinal herb for the treatment of stroke. Historically, natural products utilized in traditional medicine have been invaluable for drug development , 2. HoweAn attempt to address this involves selecting all of the medicinal herb combinations that exist within each prescription bundle and determining the frequency of these combinations. Despite the loss of individual customizability, selection of the highest frequency medicinal herb combinations may constitute a candidate group for the development of a new prescription for universal application.Dongeuibogam (dong yi bao gian),\u201d a principal piece of Korean medicine literature [Therefore, in this study, after selecting all of the prescriptions for the treatment of stroke (PTSs) recorded in \u201cterature , the freThe rationale for selecting multiple medicinal herb prescriptions, as opposed to single medicinal herbs, is as follows: (1) traditional East Asian medicines are typically available in the form of prescriptions ; (2) preWhile several previous studies have analyzed the frequency of medicinal herb combinations for various investigatory purposes \u201312, thisThis study comprised three steps. Each step was performed as described in the following paragraphs.Pung chapter\u201d in \u201cDongeuibogam,\u201d their indications were analyzed and the medicinal herbs constituting each of the PTSs were selected.In the first step, after selecting all of the prescriptions recorded in the \u201cIn the second step, the combinations with the highest repeat frequencies were selected as candidates of a medicinal herb combination for the treatment of stroke (CMHCTS), and all medicinal herbs which comprise these combinations were selected as candidates of a medicinal herb for the treatment of stroke (CMHTS). Only the medicinal herbs with doses in the upper 80% cumulative proportion per prescription were included in the CMHCTS. This ensured that only main therapeutic medicinal herbs were selected.This methodology assumed that the higher the dose within a prescription, the stronger the effect and that the more frequently a medicinal herb is included in prescriptions to treat symptoms, the more important it is . in vitro studies, in vivo studies, clinical studies, and reviews and then analyzed again for research performance status.We searched for 13 CMHTSs in the previous studies and identified relevant studies. Next, studies were specifically divided intoIn addition to commonly used scientific databases , Korean databases were used since we were searching specifically for studies related to Korean medicine (KM). The starting period for these study searches was not defined; however, December 31, 2014, was set as the final time point.We used the following terms for the searches: \u201cScientific names of CMHTS + stroke, cerebral ischemia, ischemia-reperfusion, middle cerebral artery occlusion, hypoxia, oxygen-glucose deprivation, neuroprotection, cerebrovascular protection, anti-neuroinflammation, blood-brain barrier\u201d.Pung chapter\u201d in \u201cDongeuibogam\u201d and each PTS comprised an average of 7.9 medicinal herbs.In total, 92 PTSs were selected from the \u201c\u2009Ba bao hui chun tang (26).\u2009Ba wei shun qi san (8).\u2009Ba wu tang jia nan xing ban xia zhi shi zhu li sheng jiang zhi (13).\u2009Bu huan jin dan (13).\u2009Chuan xiong shi gao san (8).\u2009Da qin jiao tang (16).\u2009Da sheng feng tang (9).\u2009Dao tan tang (7).\u2009Di huang yin zi (15).\u2009Di tan tang (10).\u2009Ding feng bing zi (9).\u2009Du shen tang jia zhu li jiang zhi (3).\u2009Er chen tang (5).\u2009Er shen dan (10).\u2009Fang feng tong sheng san (18).\u2009Huan gu dan (16).\u2009Huo ming jin dan (15).\u2009Huo xiang zheng qi san jia nan xing mu xiang fang feng dang gui (17).\u2009Jia jian dao tan tang (17).\u2009Jia jian pai feng tang (17).\u2009Jia jian run zao tang (23).\u2009Jia jian xu ming tang (13).\u2009Jia wei da bu tang (23).\u2009Jia wei jing zhou bai yuan zi (8).\u2009Jie yu wan (8).\u2009Jing zhou bai yuan zi (4).\u2009Li qi qu feng san (17).\u2009Long xing dan (12).\u2009Mi chuan shun qi san (18).\u2009Mu xiang bao ming dan (26).\u2009Niu huang ding zhi wan (14).\u2009Niu huang qing xin yuan (30).\u2009Pai feng tang (14).\u2009Pi xun ding zi (19).\u2009Pi yue wan (6).\u2009Qian zheng san (3).\u2009Qiang huo yu feng tang (28).\u2009Qin jiao sheng ma tang (10).\u2009Qing qi xuan feng san (21).\u2009Qing shen jie yu tang (19).\u2009Qing tan shun qi tang (14).\u2009Qing xin san (9).\u2009Qing yang tang (10).\u2009Qu feng chu shi tang (19).\u2009Qu feng dan (1).\u2009Qu feng zhi bao dan (26).\u2009Quan sheng hu gu san (8).\u2009Ren shen qiang huo san (16).\u2009Ren shen shun qi san (14).\u2009San he tang (11).\u2009San sheng yin (5).\u2009Shen li tang (16).\u2009Shen xiang ban xia tang (6).\u2009Shi quan da bu tang (12).\u2009Shu feng shun qi tang (22).\u2009Shu feng shun qi yuan (12).\u2009Shu feng tang (14).\u2009Shu jin bao an san (15).\u2009Si bai dan (20).\u2009Si jun zi tang jia er chen tang zhu li sheng jiang zhi bai jie zi (12).\u2009Si jun zi tang jia zhu li sheng jiang zhi (6).\u2009Si jun zi tang (4).\u2009Si wu tang jia zhu li sheng jiang zhi fu zi wu tou (8).\u2009Si wu tang jia zhu li sheng jiang zhi tao ren hong hua bai jie zi (9).\u2009Si wu tang jia zhu li sheng jiang zhi (6).\u2009Si wu tang (4).\u2009Su he xiang yuan (15).\u2009Su jing yuan (19).\u2009Tian tai san (19).\u2009Tian xian gao (4).\u2009Tie tan yuan (5).\u2009Tong qi qu feng tang (12).\u2009Tou bing dan (12).\u2009Wan jin tang (14).\u2009Wu long dan (4).\u2009Wu yao shun qi san (12).\u2009Xi jiao sheng ma tang (9).\u2009Xiao tong sheng san (12).\u2009Xiao xu ming tang (13).\u2009Xie she bai yuan zi (8).\u2009Xing fu san (9).\u2009Xu ming zhu san (15).\u2009Yang rong tang (20).\u2009Yi li jin dan (11).\u2009Yu feng dan (13).\u2009Yue bi tang (6).\u2009Yun qi san (10).\u2009Zheng she san (3).\u2009Zhi bao dan (11).\u2009Zhuan she gao (11).\u2009Zi run tang (10).\u2009Zi shou jie yu tang (10).Chinese names followed by the number of constituents are as follows:The following medicinal herb combinations were selected: 82 combinations of one medicinal herb, 581 combinations of two medicinal herbs, 2078 combinations of three medicinal herbs, 5691 combinations of four medicinal herbs, 12,522 combinations of five medicinal herbs, 22,086 combinations of six medicinal herbs, and 31,335 combinations of seven medicinal herbs. By focusing on the top five of each of these (plus ties), selection of the following occurred: six combinations comprising one medicinal herb, five combinations of two medicinal herbs, 13 combinations of three medicinal herbs, seven combinations of four medicinal herbs, six combinations of five medicinal herbs, 19 combinations of six medicinal herbs, and three combinations of seven medicinal herbs. These comprised the CMHCTS with the highest probability of efficacy in the treatment of stroke .A total of 1,494 studies of 13 CMHTSs were found; of these, 103 studies were concerned with effects in stroke, resulting in an average of 7.9 publications per candidate herb .Dongeuibogam\u201d by analyzing frequency and effectiveness. Then, analysis of the previous studies has been done.In this paper, medicinal herbs which have high possibility of stroke treatment effect in KM were selected from \u201c Angelica gigas Nakai, root: vanillic acid (VA) obtained naturally from the plant Angelica sinensis improves spatial learning and memory retention by preventing oxidative stress; (2) Ostericum koreanum (Max.) Kitagawa, rhizome: Ostericum koreanum has vasodilation effect via change of brain bloodstream; (3) Arisaema amurense Maximowicz, rhizome X: none; (4) Atractylodes japonica Koidzumi, rhizome: Atractylodes japonica Koidzumi prevent the growth inhibition, mitochondrial injury, and apoptosis of neurons induced by hypoxia; (5) Fraxinus rhynchophylla Hance, cortex X: none; (6) Gastrodia elata Bl., rhizome: Gastrodia elata attenuate the hippocampal neuronal damage and decrease necrosis; (7) Glycyrrhiza uralensis Fisch., root: Glycyrrhiza uralensis Fisch. has neuroprotective efficacy in the postischemic brain via its anti-inflammatory, antiexcitotoxic, and antioxidative effects; (8) Ligusticum chuanxiong Hort., rhizome: Ligusticum chuanxiong Hort. reduces cerebral infarct through its antioxidative and anti-inflammatory effects; (9) Paeonia lactiflora Pallas, root: paeoniflorin may play the role of antagonising cerebral ischemia by adjusting cerebral energy metabolism and nitric oxide formation; (10) Pinellia ternata (Thunb.) Breit., rhizome X: none; (11) Poria cocos Wolf, sclerotium: Poria cocos have neuroprotective effects against the acute restriction of metabolite and oxygen supply in cerebral blood flow; (12) Rehmannia glutinosa Liboschitz, root: Catalpol, an iridoid glycoside abundant in the roots of Rehmannia glutinosa, exerts the cytoprotective effect on astrocytes by suppressing the production of free radicals and elevating antioxidant capacity; (13) Scutellaria baicalensis Georgi, root: Scutellaria baicalensis Georgi dramatically reduce the decrease in learning and memory, attenuated neuronal injury, and improved abnormality of energy metabolites.Look at the possible mechanisms of 13 CMHTSs in To sum up, stoke treatment by antioxidative effect and anti-inflammatory effect was mostly common. There were many research papers about neuroprotective effect by energy metabolism and controlling blood circulation as well. Angelica gigas and 3 for Ligusticum chuanxiong) among 103 previous studies. Simply look at the result: (1) Angelica gigas Nakai, root: Angelica injection has evident therapeutic effect in treating acute cerebral infarction.; (2) Ligusticum chuanxiong Hort., rhizome: Ligusticum chuanxiong and its effective components improve brain microcirculation through inhibiting thrombus formation and platelet aggregation as well as blood viscosity.In addition, there are only 4 clinical studies \u201cDongeuibogam\u201d is the comprehensive summary of all the traditional medicines of Northeast Asia prior to the 17th century, because it is based on rigorous selection of 189 of the major medicinal literature sources of the region [There are three main reasons for selecting \u201ce region ; (2) it e region ; (3) excPung chapter\u201d specializes in symptom which is the most similar to symptom of today's stroke. Therefore, it is appropriate to match today's stroke and stroke written in the classical literature to select CMHTSs.Third of all, you may wonder if it is possible to match today's stroke and stroke written in the classical literature. Even though definition of stroke in Korean traditional and Western medicine is slightly different, \u201c Zingiber officinale Rosc. and Zizyphus jujuba var. inermis Rehder, the so-called \u201csh\u01d0 y\u00e0o\u201d are added a little for balance of medicinal herbs or to improve digestive functions. These \u201csh\u01d0 y\u00e0o\u201d do not have major treatment effect but are frequently added in prescriptions, which means that frequently used medicinal herbs in prescription do not mean the herbs are principle ingredients. And therefore, the minor herbs were excluded from CMHCTS and only 80% of medicinal herbs in PTS were included in CMHCTS. The other doubt in the second step of method is that, instead of selecting the most frequently used medicinal herbs in 94 PTSs as CMHTS, why CMHTS is selected after sorting CMHCTS out. The reason is that prescriptions are not simply a quantitative addition of the individual medicinal herbs; instead they produce a superior efficacy to single medicines [Fourth of all, you may wonder why 80% of medicinal herbs in PTS are only included in CMHCTS in the second step of method. In Korean traditional prescription, such asLastly, you may wonder about necessity of third step of method and result of the step, The fundamental questions discussed above are not only key point but also character of this paper. In conclusion, methodology used in this study is regarded as meaningful challenge to discover \u201ca hidden treasure\u201d for stroke from classical literature. And the result of this study, some CMHCTSs and 13 CMHTSs, will be certainly valuable as fundamental data for experiment and clinical research.Dongeuibogam\u201d and reviewed the results of previous studies regarding the effects in stroke. In order to develop a universally applicable PTS, it will be necessary to conduct longer and more complex experiments and clinical trials. However, the CMHCTSs and CMHTSs proposed in this study have the potential to reduce the experimental and developmental time period. Furthermore, this study demonstrates the utilization of text mining for the development of universally applicable prescriptions for a particular disease.In the present study, we finally selected some CMHCTSs and 13 CMHTSs from the \u201c"} +{"text": "Preoperative diagnostics of ovarian neoplasms rely on ultrasound imaging and the serum biomarkers CA125 and HE4. However, these markers may be elevated in non-neoplastic conditions and may fail to identify most non-serous epithelial cancer subtypes. The objective of this study was to identify histotype-specific serum biomarkers for mucinous ovarian cancer. The candidate genes with mucinous histotype specific expression profile were identified from publicly available gene-expression databases and further in silico data mining was performed utilizing the MediSapiens database. Candidate biomarker validation was done using qRT-PCR, western blotting and immunohistochemical staining of tumor tissue microarrays. The expression level of the candidate gene in serum was compared to the serum CA125 and HE4 levels in a patient cohort of prospectively collected advanced ovarian cancer. Database searches identified REG4 as a potential biomarker with specificity for the mucinous ovarian cancer subtype. The specific expression within epithelial ovarian tumors was further confirmed by mRNA analysis. Immunohistochemical staining of ovarian tumor tissue arrays showed distinctive cytoplasmic expression pattern only in mucinous carcinomas and suggested differential expression between benign and malignant mucinous neoplasms. Finally, an ELISA based serum biomarker assay demonstrated increased expression only in patients with mucinous ovarian cancer. This study identifies REG4 as a potential serum biomarker for histotype-specific detection of mucinous ovarian cancer and suggests serum REG4 measurement as a non-invasive diagnostic tool for postoperative follow-up of patients with mucinous ovarian cancer. Epithelial ovarian cancer (EOC) is the most lethal gynecological cancer . AlthougThe diagnosis of ovarian cancer relies on clinical features, ultrasound imaging and serum biomarker Cancer antigen 125 (CA125). In addition, the novel serum biomarker Human epididymis 4 (HE4) may bring additional information in differential diagnostics of women with pelvic mass . Since tin silico analyses, we studied the expression of Regenerating Islet-Derived Protein IV (REG4) in a set of mucinous ovarian cancer tissues, patient sera and ascites fluids. In addition, REG4 concentration was measured in a time series of serum samples to determine the correlation to tumor burden. Furthermore, we examined CA125, HE4 and REG4 in the same samples to compare their performance.This study aimed at identification of mucinous histotype specific serum biomarkers for ovarian cancer. Based on www.medisapiens.com) was applied to study the gene expression levels across all normal and neoplastic human tissues [The MediSapiens database was constructed from surgical specimens collected from the tissue archive of Department of Pathology, Turku University Central Hospital as previously described . The useThe TMA contained 185 samples from 100 patients with ovarian neoplasms; serous cystadenomas (n = 16), serous borderline tumors (n = 27), serous carcinomas (n = 47), mucinous cystadenomas n = 14), mucinous carcinomas (n = 36), endometroid carcinomas (n = 45). The TMA was stained with mouse monoclonal REG4 antibody , as desc, mucinou\u0394\u0394CT method (Applied Biosystems). Three or more replicate samples were used to study the mRNA expression of REG4 and GAPDH. Average expression of the control samples was considered for the calculation of the fold changes and GAPDH was used as an endogenous control. Three or more replicate samples were studied for detection of mRNA expression. Statistical significance was determined with student\u2019s t-test, and p-values < 0.05 were considered statistically significant.Total RNA was extracted from frozen ovarian tumor samples (10 mg piece) and primary cells using RNeasy Mini kit (Qiagen) according to the manufacturer\u2019s protocol and processed to cDNA . TaqMan probes were acquired from the Universal Probe Library and primers were ordered from Oligomer for REG4 and GAPDH . Real-time quantitative RT-PCR (qRT-PCR) was done with ABI Prism 7900 (Applied Biosystems). Quantitation was carried out with RQ manager 1.2 software using the For protein lysates, approximately 20 mg pieces were cut from frozen tumors and lysed with sonication in RIPA lysis buffer. Protein concentrations were measured using BioRad protein assay kit according to manufacturer\u2019s instructions . Samples were prepared mixing appropriate amounts of protein lysate with 5 x Laemmli buffer and boiled for 5 min. Equal amounts of protein (20 \u03bcg) were separated on 15% SDS-PAGE gel and transferred to nitrocellulose membrane . Equal loading was confirmed with Ponceau staining. Membranes were blocked with 5% milk-TBST and probed with polyclonal REG4 antibody and HRP-conjugated anti-goat secondary antibody . Bound proteins were detected by enhanced chemiluminescence.The REG4 sandwich ELISA assays were performed using the Human REG4 ELISA Pair Set according to the manufacturer\u2019s instructions. The serum samples were diluted 1:1000 and ascites samples 1:500 in the sample dilution buffer. A pool of serum samples from healthy males (n = 4) was used as a negative control. The absorbance at 450 nm was measured with Victor 1420 Multilabel Counter . All controls and samples were analyzed in two replicate wells in three individual runs.in silico analysis of gene expression data using the GTI algorithm [in silico expression data as well as relevant literature was conducted. The final candidate selection identified the Regenerating Islet-Derived Protein IV (REG4) as a potential serum biomarker for mucinous ovarian cancer histotype. REG4 is a member of the regenerating gene (REG) family, which belongs to the calcium-dependent lectin superfamily. It consists of 17 members, from which REG1, REG2, REG3 and REG4 have been associated with inflammation, diabetes and cancers [In order to identify putative serum biomarker candidates for ovarian cancer detection and histotype specification, we performed lgorithm and the lgorithm . The analgorithm ), after REG4 mRNA expression levels were studied in more detail using the MediSapiens database which includes expression data of human tissue samples than in healthy tissues (n = 3082). The results are summarized in REG4 mRNA was found in malignant gastric, colorectal and pancreatic tissues as well as in healthy colorectal and pancreatic tissues. The highest level of REG4 mRNA expression was seen in mucinous ovarian cancer, while the expression level remained very low in healthy ovarian tissue. More specific analysis of REG4 mRNA expression in healthy ovary (n = 10) and malignant (n = 288) ovarian tissues of various histologies, confirmed the mucinous histotype specific expression profile and high-grade serous (SER) ovarian tumors. The REG4 mRNA expression was studied with qRT-PCR using mRNA from serous tumors as negative control. The results are summarized in REG4 was not expressed in mucinous cystadenoma (MUC_A_1) and showed significantly lower mRNA expression levels also in mucinous borderline tumors (MUC_B_2\u20133). REG4 protein expression in the same set of tumor samples was assessed with western blotting. The results indicated high REG4 protein levels in REG4 mRNA positive tumors whereas the REG4 concentration remained below the cut-off value (FC < 1) . In the (FC < 1) . Both saIn order to determine the correlation of REG4, CA125 and HE4 concentrations to tumor burden, serum samples were measured at multiple time points from a patient (M1) treated for disseminated mucinous carcinoma see showed tOvarian neoplasms differ in their genetic profiles, biological behavior and outcome, and should be treated as distinct diseases already during surgical intervention. Thus, a specific preoperative diagnosis is crucial for selection of optimal treatment strategy. Currently there are no methods for preoperative discrimination of ovarian cancer subtypes and the specific diagnosis is reached only by perioperative frozen section or postoperative histopathological analysis. Here we suggest REG4 as a promising serum biomarker for separating mucinous cancers from other epithelial ovarian cancer subtypes. According to our results, serum REG4 analysis may also have potential for differentiation between benign and malignant mucinous neoplasms and for follow-up of mucinous ovarian cancer patients.in silico analyses, which indicated very specific mRNA expression profile of REG4 in mucinous cancers among various ovarian tumor subtypes. REG4 expression was further validated with qRT-PCR and western blotting from fresh frozen ovarian tumor tissue samples originating from serous and mucinous subtypes. As expected based on the in silico data, REG4 was highly expressed only in malignant mucinous ovarian tumors. Finally, immunohistochemical staining of a tumor tissue microarray consisting of 185 tumor cores from benign and malignant serous, endometroid and mucinous neoplasms confirmed restricted expression in the mucinous lineage. Interestingly, no REG4 expression was detected in mucinous cystadenomas suggesting that immunohistochemical REG4 analysis could be used as an additional method for distinction between benign and malignant mucinous ovarian tumors. All the mucinous samples were confirmed to be of primary ovarian tumor origin. Our sample set did not contain specimens of the more rare types of mucinous ovarian carcinoma; such as endocervical/ seromucinous or foveolar types. Therefore, in order to determine the true potential of REG4 as an immunohistochemical marker, additional analyses with wider range of different types of samples are needed.The specific mucinous lineage expression of REG4 within ovarian neoplasms was demonstrated by several ways. The first evidence was obtained from The serum biomarkers CA125 and HE4 are used for ovarian cancer diagnostics and follow-up. These markers mainly detect serous cancers ,19, but REG4 has been previously indicated as a serum biomarker in gastric, colorectal and prostate cancer \u201324. ThusThe function of REG4 in ovarian cancer is poorly understood and has not been widely investigated. A recent study suggests that REG4 modulates proliferation, apoptosis, migration and invasion of ovarian cancer cells, and has an important role in early ovarian carcinogenesis . This stIn conclusion, we identified REG4 as a potential biomarker for subtype specific diagnosis of mucinous ovarian cancer as well as for disease follow-up. According to our results REG4 protein is secreted into circulation by mucinous ovarian tumor cells and can be detected with an ELISA-based test. We measured three biomarkers from patient sera; CA125, HE4 and REG4 and found high concentrations of REG4 in preoperative samples of mucinous ovarian cancer patients, whereas CA125 and HE4 failed to convincingly detect malignancy in these samples. Our study suggests that combining REG4 with CA125 and HE4 could provide added value for preoperative diagnostics of ovarian masses of potential neoplastic origin, and enable distinction of mucinous ovarian cancer from other ovarian cancer subtypes with non-invasive methods.S1 Table(XLSX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.S4 Table(DOCX)Click here for additional data file.S1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file."} +{"text": "In acute ST-elevation myocardial infarction (STEMI) oxidative stress is an important determinant of severe myocardial damage and reperfusion injury. However, no data are currently available about the correlation of oxidative stress with cardiac magnetic resonance (CMR) markers of myocardial injury in acute STEMI.We studied 198 patients with acute STEMI undergoing primary percutaneous coronary intervention (PCI) within 12 hours of symptom onset. Oxidative stress was determined by oxidated LDL-cholesterol (oxLDL), activated oxygen protein products (AOPP) and lipid hydroperoxides (perOx). Blood samples were collected before PCI, after PCI and on day 2 as well as day 3 and 4. Follow-up values were expressed as ratio to baseline values. CMR studies were performed 2-4 days after the infarction using a standard infarction protocol.A rise in oxidative stress parameters after reperfusion was found in oxLDL for 50.2%, in AOPP for 38.3% and in perOx for 55.2% of all STEMI patients on day 2. There were no significant differences in CMR parameters when comparing patients according to the rise or fall of oxidative stress parameters, except for microvascular obstruction (MO) and AOPP (p=0.043, Table Rise or fall of markers of oxidative stress had no relevant impact on CMR markers of myocardial damage in acute STEMI. Further studies are necessary to elucidate the exact detrimental effect of oxidative stress on myocardial injury in acute reperfused STEMI.N/A."} +{"text": "HIV-1 is primarily a mucosal pathogen since more than 80% of infections occur through genital exposure. Vaginal intercourse, though an inefficient mode of transmission, contributes more new infections worldwide than any other route. Also, the female reproductive tract has been identified as a compartment that harbors variants distinct from blood. Present study aims to highlight viral compartmentalization between these compartments reflecting inadequate ART penetration.In this study blood and cervicovaginal swabs were collected from 8 female subjects. CD4 counts and viral loads were determined. Translated amino acid sequences of C2 v3 region of env gene of proviral HIV-1 C in PBMCs and genital cells were analyzed using N-Glycosite and Geno2pheno [Co receptor] 1.2 programs for presence of N linked glycosylation sites and co receptor preference, respectively.env gene of HIV-1 C shows variation in the number of N linked glycosylation (NLG) sites and uniform co receptor preference. Viral load varies in blood and genital secretions.Characterization of translated amino acid sequences of C2 v3 region of Genotypic characterization of viral variants in blood and female reproductive tract can provide information regarding their association with sexual transmission of HIV. Difference in the number of NLG sites observed may influence the affinity for host cell co receptor. Discrepancies in viral load of blood and genital secretions suggest that ART may not be uniformly effective in suppression of viral load in different compartments of the same individual."} +{"text": "Rapid correction of hyponatremia can cause pontine myelinolysis and should be corrected by less than 10 mM/L within 24 hours. Hepatic cirrhosis is often associated with hyponatremia due to fluid retention and ascites. During liver transplantation, sodium may increase because of fluid administration and blood transfusions. Little is known regarding the frequency of rapid sodium increases during liver transplantation and whether such rapid corrections carry detrimental effects.To determine the incidence of rapid plasma sodium correction in liver transplants and their effect on outcome.We retrospectively analyzed data from 607 liver transplants at a single institution. Rapid correction of sodium was defined as an increase of serum sodium by more than 10 mM/L within 24 hours after the start of liver transplantation when compared to preoperative sodium levels. Primary endpoint was 90-day mortality or graft failure requiring re-transplantation.Plasma sodium concentrations increased by more than 10 mM/L within 24 hours after liver transplantation in 39 of 607 patients (6.4%). These patients had lower preoperative sodium levels and higher MELD scores . Sodium levels increased by 14.6 +/- 3.9; range 11-28 mM/L in the rapid increase group of the patient described above are more likely attributable to her preoperative acute liver failure than rapid increase of sodium concentrations."} +{"text": "Endomyocardial biopsy (EMB) is widely used for routine surveillance of cardiac allograft rejection. The need for continued EMB beyond the first year after cardiac transplantation is controversial.The aim of this study was to investigate the use of EMB in monitoring long term surviving heart transplant recipients.We conducted a retrospective chart review of all patients at our center 2 years or more after heart transplantation. 154 HTx patients between 2000-2012 were included in this study. Significant cellular rejection was defined as grade 2R or 3R using ISHLT nomenclature. Patients were analyzed assessing immunosuppressive regimen and procedural related complications.Of 154 cardiac transplant patients, 110 (71.4%) had a follow-up of more than 2 years. Interestingly, 17 of these long-term survivors of cardiac transplantation developed at least 1 episode of significant late (>2 years after Tx) cellular rejection (15.5%). Analyzing the respective immunosuppressive regimen showed increased number of calcineurin inhibitor (CNI)-free regimen (64.7%) in patients rejecting late after heart transplantation. The overall incidence of procedural related complications was low (1.0%) and none was life-threatening.The above data demonstrates that endomyocardial biopsies continue to detect clinically significant rejection beyond 2 years after cardiac transplantation. Late rejection was not depending on previous episodes of early cellular rejections. Therefore, we recommend long-term routine endomyocardial biopsies in cardiac transplant recipients especially in patients at high risk due to the immunosuppressive regimen."} +{"text": "Inflammatory myopathies during childhood are clinically, biologically and pathologically heterogeneous.The objective of this study is to collect cases of pediatric-onset overlap myositis to improve description and classification.Retrospective study of patients followed in Necker-Enfants Malades Hospital from january 2002 to march 2014, with overlap myositis defined by the association of inflammatory myositis and clinic features and/or biological signs of other connective tissue diseases . For every patient we collected at diagnosis clinical features, biological profile, histological and radiological (MRI) data, treatments and evolution.Twenty patients were included, 3 boys and 17 girls. The median age was 9 years 5 months (age between 3 years 9 months and 14 years 1 month). Every patient had myositis at diagnosis associated with polyarthritis (7 patients), Raynaud syndrome (4 patients), esophageal dyskinesia (2 patients), lung damage . Two patients had sclerodactyly, 5 had morphea. One patient had typical dermatomyositis at diagnosis and developed pulmonary involvement and cutaneous sclerosis during evolution. Serum creatine kinase level was elevated for thirteen patients at diagnosis. Eighteen patients had autoantibodies at diagnosis or during evolution . Muscular MRI was performed on 12 patients . Muscular biopsy was done on 16 patients . The first-line treatment included corticosteroid therapy (18 patients), alone (11 patients) or associated with hydroxycholoroquine (8 patients), methotrexate (5 patients), intravenous immunoglobulins (2 patients), mycophenolate mofetil (2 patients), azathioprine (1 patient) and plasmatic exchange (1 patient). The median duration of follow up is 20 months (from 7 months to 5 years): no patient died. Ten patients are in complete remission on the muscular plan. Nevertheless a majority develops a connective tissue disease .This is the first study emphasizes juvenile overlap myositis. Juvenile overlap myositis are heterogeneous diseases and differ from adult myositis. A specific pediatric classification is warranted to improve characterization and to adapt treatments. MRI and histological datas appear to be relevant, nevertheless it is essential to evaluate their reliability prospectively so that we can use them to improve classification and determination of prognostic factors.None declared."} +{"text": "Background. Intra-articular fractures of distal femur present a huge surgical challenge. The aim of this study is to evaluate functional outcome, fracture healing, and the complications of distal femoral intra-articular fractures using locking compression plates. Material and Methods. We reviewed 46 distal femoral fractures treated with distal femoral locking compression plates between 2009 to 2012. There were 36 men and 10 women with mean age of 35 years (range 20\u201372). More than half of the patients were of type C3 (AO classification) and had been caused by high energy trauma with associated injuries. Results. 2 patients were lost to follow-up. Of the remaining 44 patients, the mean follow-up period was 25 months (range 18\u201336). The mean time for radiological union was 12 weeks (range 10\u201318) except 2 patients which had gone for nonunion. At the latest follow up ROM >120\u00b0 is noted in 32 patients, 90\u2013120 in 10 patients, and 70\u201390 in 2 patients. 38 patients (86%) had good/excellent outcome. Conclusion. Use of standard lateral approach for simple intra-articular distal femoral fractures (C1) and transarticular/minimally invasive techniques for complex intra-articular fractures (C2/C3) results in improved exposure of the knee joint and better union rates with low incidence of bone grafting. Intra-articular fractures of distal femur present a huge surgical challenge. These fractures are difficult to treat and operative treatment is usually recommended for favorable outcome \u20134. TheseCurrent generation of distal femoral locking compression plates is precontoured based on the average bony anatomy of the adult population and they form a fixed angled construct. The pull-out strength of locking screws is higher than the conventional screws and is particularly useful in osteoporotic bones. These plates are designed to apply in minimally invasive fashion to preserve local biology and avoid problems with fracture healing and infection , 7.The purpose of this study was to evaluate functional outcome, fracture healing, and the complications of distal femoral intra-articular fractures using locking compression plates.We reviewed 46 consecutive distal femoral fractures treated with distal femoral locking compression plates between 2009 and 2012. AO Synthes distal femoral locking compression plate and Zimmer periarticular distal femoral locking plate were used in 19 and 27 patients, respectively. There were 36 men and 10 women with mean age of 35 years (range 20\u201372).Fractures were categorized according to AO/OTA classification. Inclusion criteria were as follows: (i) type C distal femoral fractures and (ii) patients older than 18 years of age. Pathological fractures and types A and B distal femoral fractures were excluded from the study.10 patients had intra-articular fractures involving both condyles (33C-1), 12 patients had metaphyseal comminution (33C-2), and 24 patients had articular comminution (33C-3). Definitive fracture fixation was performed in 33 patients within 7 days. 13 patients with multiple injuries and open fractures were treated within 3\u2009wks after temporary application of external fixator. Preoperative AP and lateral radiographs of affected knee with femur were obtained. CT was obtained in 30 patients.All surgeries were carried out at our tertiary care level 1 trauma centre with patient placed supine on the radiolucent table. A small towel bump was placed posterior to supracondylar region. A femoral distractor was used in 15 patients. Lateral approach was used for C1 fractures. Iliotibial band was incised along the line of skin incision. Vastus lateralis was retracted anteriorly to expose the distal femur. Anterolateral parapatellar approach was used in C2 and C3 fractures. Intra-articular fracture reduction was obtained and temporarily fixed with multiple K wires. Indirect reduction of articular surface with femoral diaphysis was done under fluoroscopic guidance. Appropriate-sized distal femoral locking plate was slid in distal to proximal direction submuscularly over lateral aspect of distal femur. Length of plate was determined intraoperatively after fracture reduction. Usually we prefer minimum length of the plate which is three times the fracture comminution segment. First a standard 4.5\u2009mm cortical screw was placed in the femoral diaphysis and tightened lightly. For proximal fixation 3 or 4 bicortical screws were used percutaneously. Minimum of 5 locking screws were used for distal fixation . One or Postoperative radiographs were taken. Follow-up radiographs were taken after 6\u2009wks, 12\u2009wks, 6 months, 9 months, and 12 months after surgery. Initially nonweight bearing mobilization was started. Gradual weight bearing was started based on the evidence of bridging callous on follow-up radiographs. The average time until weight bearing was 3 months. Radiological union was defined as the presence of cross trabeculation on both AP and lateral radiographs. Nonunion was defined as failure of fracture union at 9-month follow up. Clinical and functional outcomes were assessed using the Knee Society score Table 2Table 2.The mean follow-up period was 12.3 months (mean 9\u201324 months). The mean time for radiological union was 14 weeks (range 8\u201318 weeks). There were 2 cases of nonunion. One case required autogenous iliac crest bone grafting and the other case required bone grafting along with refixation using longer plate due to breakage of proximal screws with broken proximal screws . FracturOne patient required arthroscopic arthrolysis with implant removal due to severe restriction of movement and hardware prominence. No cases of infection and rotational or angular deformity more than 5 degrees was noted . 2 patieAt the latest follow up ROM > 120\u00b0 is noted in 32 patients, 90\u2013120 in 10 patients, and 70\u201390 in 2 patients.To maintain the fracture biology and to minimize the soft tissue trauma, minimally invasive plating techniques have been developed for the fixation of distal femoral fractures.The main goals of the above-mentioned techniques are to maintain the important anatomy and to promote early fracture healing.Locking plate systems such as the LISS have beePrevious studies have demonstrated successful early results and relatively low complication rates using minimally invasive plating techniques for the fractures of distal femur , 17, 18.We have used minimally invasive plate fixation technique using standard lateral approach for the fixation of simple intra-articular fractures (C1). However, more extensive approaches are needed for fixation of complex intra-articular fractures (C2/C3). In these fractures we have employed lateral parapatellar arthrotomy for direct reduction of joint surface. This articular block was fixed to the femoral shaft using indirect plate fixation technique.In our series of 44 patients, there were no cases of infection and varus/valgus alignment of more than 5 degrees. We noted 2 cases of nonunion, out of which one case required autogenous iliac crest bone grafting and the other case required bone grafting along with refixation using longer plate due to breakage of proximal screws. One case required arthrolysis with implant removal due to severe restriction of ROM and hardware prominence.As reviewed by Miclau et al. bone graEarly experience with the LISS for distal femoral fractures in multicentric study in Europe demonstrated a 20% incidence of varus/valgus deformity greater than 5 degrees [Limitations of the study include (i) all the drawbacks pertaining to its retrospective nature and (ii) small sample size.Strength of our study is that we included only intra-articular fractures; most of these were complex which require adequate surgical expertise.The results of our study suggest that use of standard lateral approach for simple intra-articular distal femoral fractures (C1) and transarticular/minimally invasive techniques for complex intra-articular fractures (C2/C3) results in improved exposure of the knee joint and better union rates with low incidence of bone grafting. However randomized trials are needed to assess this technique."} +{"text": "Investigators from New York Presbyterian Morgan Stanley Children's Hospital examined whether having an isolated headache following minor blunt head trauma was suggestive of traumatic brain injury (TBI) among a large cohort of children 2-18 years of age. Investigators from New York Presbyterian Morgan Stanley Children's Hospital examined whether having an isolated headache following minor blunt head trauma was suggestive of traumatic brain injury (TBI) among a large cohort of children 2-18 years of age [COMMENTARY. Head trauma in children results in 600,000 emergency room visits per year. The decision to obtain a CT must be balanced against its associated risks. Radiation from CT scans can cause malignancy in children and the risk has an inverse relationship with age. The PECARN prediction rules identified children at very low risk of ciTBI after blunt head trauma for whom CT scans might be avoided. Several studies report that less than 10% of CT scans show TBIs in children with minor head trauma , 3. Addi"} +{"text": "Please download this article again to view the correct version. The originally published, uncorrected article and the republished, corrected article are provided here for reference.The article titled \"GDP versus ESHAP Regimen in Relapsed and/or Refractory Hodgkin lymphoma: A Comparison Study\"S1 File(PDF)Click here for additional data file.S2 File(PDF)Click here for additional data file."} +{"text": "Five major candidate genes known to be associated with syndromic and non-syndromic CTA, these are PAX9, MSX1, AXIN2, EDA and WNT10A. The present investigation was undertaken to identify and characterize disease causing genetic variant by conventional genotyping and whole genome sequencing in familial tooth agenesisCongenital tooth agenesis (CTA), a type of craniofacial disorders affects approximately 20% (including 3We have identified two Indian families (DEN11 and DEN12) segregating two distinct types of non-syndromic tooth agenesis with autosomal dominant (DEN11) and apparently indistinguishable either autosomal or X-linked dominant pattern of inheritance (DEN12) family. Eight affected and four unaffected members from DEN11 and four affected and two unaffected members from DEN12 family were enrolled for the present study to identify causative genetic defect associated with the disease. While direct DNA sequencing and microsatellite based genotyping were carried out to screen all but EDA genes in affected and unaffected family members of DEN11, whole genome sequencing was carried out for two affected (III-2 and IV-4) family members from DEN12 along with four unaffected unrelated controls using Illumina 2500 Next Generation Sequencing platform for identification and subsequent characterization of causative variant/s.In the microsatellite and SNP based haplotype analysis we identified a haplotype block of a ~9.64 Mb region containing PAX9 gene located between D14S70 and D14S288 markers associated with CTA in DEN11. However, absence of any DNA sequence variant within the exons and exon-intron boundaries of the linked PAX9 was found and this indicates the involvement of other pathogenic mechanism. In the whole exome sequencing we identified 86 nonsynonymous novel nucleotide variations distributed among 84 different genes across the nuclear genome of two affected members of DEN12 subjected for investigation. Using bioinformatics, among those variations a specific variation in Ectodysplasin-A (EDA), c.956G>T transversion leading to p.Ser319Ile, at the Tnf homology domain, was considered as a potential pathogenic variation. This variation was not observed in any unaffected family member and in 100 unrelated control chromosomes as assayed through Sanger sequencing and/or RFLP. In silico analysis using SwisSPdb Viewer V4.1.0 reveals that this change destroys an H-bond between p.319 Ser and p.332 Cys establishing this as a plausible pathogenic variation for tooth agenesis in DEN12 family."} +{"text": "Understanding how extremely low-frequency magnetic fields (MF) interact with human brain activity is an important question, especially regarding potential effects of power-lines MF (60 Hz in North America). Such knowledge is critical to 1) contribute to guidelines protecting public and workers from exposure to ELF MFs ; and 2) We used an extensively validated neural mass model describiSimulated EEG alpha power decreased with increased 60 Hz MF flux density , without significant effects from synaptic plasticity processes. If slow inhibitory interneurons were alsin silico effects of transcranial alternating current stimulation and magnetic stimulation (tACS/TMS). Future work will include frequency-dependent effects from extracellular medium dielectric properties, and selective modulation of specific neuronal populations.The model will be used to 1) understand human data currently acquired in our group ; and 2)"} +{"text": "Recombinant human IgM22 (rHIgM22) binds to myelin and to oligodendrocytes, and promotes remyelination in a mouse model of multiple sclerosis . rHIgM22"} +{"text": "CXCL14, ITGAX, LPCAT2, RNASEH2A, and ZNF322) were positively correlated with aggressive prostate cancer and two genes (CCL19 and HIST1H1A) were protective for aggressive prostate cancer. Higher than average levels of expression of the five genes that were positively correlated with aggressive disease were consistently associated with patient outcome in both human prostate cancer tumor gene expression datasets. Second, three of these five genes harbored polymorphisms associated with aggressive disease development in a human GWAS cohort consisting of 1,172 prostate cancer patients. This study is the first example of using a systems genetics approach to successfully identify novel susceptibility genes for aggressive prostate cancer. Such approaches will facilitate the identification of novel germline factors driving aggressive disease susceptibility and allow for new insights into these deadly forms of prostate cancer.Although prostate cancer typically runs an indolent course, a subset of men develop aggressive, fatal forms of this disease. We hypothesize that germline variation modulates susceptibility to aggressive prostate cancer. The goal of this work is to identify susceptibility genes using the C57BL/6-Tg(TRAMP)8247Ng/J (TRAMP) mouse model of neuroendocrine prostate cancer. Quantitative trait locus (QTL) mapping was performed in transgene-positive (TRAMPxNOD/ShiLtJ) F2 intercross males (n\u200a=\u200a228), which facilitated identification of 11 loci associated with aggressive disease development. Microarray data derived from 126 (TRAMPxNOD/ShiLtJ) F2 primary tumors were used to prioritize candidate genes within QTLs, with candidate genes deemed as being high priority when possessing both high levels of expression-trait correlation and a proximal expression QTL. This process enabled the identification of 35 aggressive prostate tumorigenesis candidate genes. The role of these genes in aggressive forms of human prostate cancer was investigated using two concurrent approaches. First, logistic regression analysis in two human prostate gene expression datasets revealed that expression levels of five genes ( Prostate cancer is a remarkably common disease, and in 2014 it is estimated that it will account for 27% of new cancer cases in men in the US. However, less than 13% those diagnosed will succumb to prostate cancer, with most men dying from unrelated causes. The tests used to identify men at risk of fatal prostate cancer are inaccurate, which leads to overtreatment, unnecessary patient suffering, and represents a significant public health burden. Many studies have shown that hereditary genetic variation significantly alters susceptibility to fatal prostate cancer, although the identities of genes responsible for this are mostly unknown. Here, we used a mouse model of prostate cancer to identify such genes. We introduced hereditary genetic variation into this mouse model through breeding, and used a genetic mapping technique to identify 35 genes associated with aggressive disease. The levels of three of these genes were consistently abnormal in human prostate cancers with a more aggressive disease course. Additionally, hereditary differences in these same three genes were associated with markers of fatal prostate cancer in men. This approach has given us unique insights into how hereditary variation influences fatal forms of prostate cancer. Prostate cancer is a common disease, and it is estimated that approximately 233,000 new cases will be diagnosed in in the United States alone in 2014 vs. 125.0 months for adenocarcinoma) One such feature that is garnering increased attention is the emergence of prostate tumors with a neuroendocrine (NE) phenotype RB1 loss is a crucial element of the pathogenesis of NE prostate cancer N-MYC and AURKAThe pathogenesis of NED remains unclear. Recent studies have demonstrated that The work presented here utilizes a systems genetics approach, which involves the integration of lines of evidence from a mouse model of aggressive prostate cancer and several human prostate cancer datasets to identify novel genes associated with aggressive disease . CandidaCXCL14, ITGAX, and LPCAT2.To explore the origins of this, an F2 mapping panel involving TRAMP and NOD/ShiLtJ, which is a strain that is highly susceptible to developing aggressive tumorigenesis, was generated. These mice were used to map quantitative trait loci (QTLs) associated with aggressive NE prostate cancer. Following this, QTL candidate genes were nominated from microarray gene expression data derived from (TRAMP \u00d7 NOD/ShiLtJ) F2 tumors through a combination of expression QTL (eQTL) mapping and gene expression-trait correlation analysis. The relevance of these QTL candidate genes to aggressive forms of human prostate cancer were explored through two concurrent approaches: first, by correlating their expression levels with disease free survival (DFS) in two prostate tumor gene expression cohorts; and second, by analyzing a human GWAS dataset to correlate the frequencies of QTL candidate gene single nucleotide polymorphisms (SNPs) with clinical markers of disease aggressiveness. This approach, which is novel to the field of prostate cancer to the best of our knowledge, facilitated the identification of three novel aggressive prostate cancer susceptibility genes: Earlier work demonstrated that germline variation present in the NOD/ShiLtJ strain renders (TRAMP \u00d7 NOD/ShiLtJ) F1 male mice significantly more susceptible to aggressive prostate tumorigenesis P\u200a=\u200a7.40\u00d710\u221211; To investigate this hypothesis, a (TRAMP \u00d7 NOD/ShiLtJ) F2 intercross population consisting of 228 transgene-positive males was developed. Mice were aged until 30 weeks of age or until humane endpoints were achieved. As expected, substantial variation in aggressive prostate cancer phenotypes was observed in these F2 mice . Of partQTLs were mapped in (TRAMP \u00d7 NOD/ShiLtJ) F2 males by performing a genome scan using 666 informative SNPs. Analyses were performed in J/qtl Integration of germline variation and transcriptome data is a well-established means of nominating QTL candidate genes that influence a given trait through expression-related mechanisms trans-eQTL). The high number of distal eQTLs, which reside on the same chromosome as the cognate transcript but outside of the 1 Mb window for mapping proximal eQTLs most likely reflects the low level of recombination typically observed in F2 populations. The genomic distributions of eQTLs relative to their cognate transcript are illustrated in To identify QTL candidate genes in this manner, microarray analysis was performed to analyze patterns of global gene expression in all available F2 prostate tumors (n\u200a=\u200a126). Expression QTL mapping was performed using Matrix eQTL To further increase the stringency of QTL candidate gene identification, the expression levels of all transcripts within the boundaries of each of 11 aggressive disease QTLs were correlated with the QTL trait \u2013S14. UsiHaving used a highly stringent analytical approach to identify 35 aggressive tumorigenesis susceptibility genes in (TRAMP \u00d7 NOD/ShiLtJ) F2 males, we aimed to determine whether the human orthologs of these genes play a similar role in human prostate cancer. Of the 35 QTL candidate genes identified in (TRAMP \u00d7 NOD/ShiLtJ) F2 males, 29 had a human ortholog . The 6 thttp://www.cbioportal.org/CXCL14 and RNASEH2A (OR \u200a=\u200a2.17 [1.04\u20134.52]) were associated with disease recurrence; and LPCAT2 (OR \u200a=\u200a1.44 [1.02\u20132.03]) with a higher pathological stage. In the GSE21032 cohort, the expression levels of three genes were associated with an increased risk of aggressive disease and two genes identified as having a protective effect. Specifically, the expression levels of CXCL14 were associated with a higher pathological stage (OR \u200a=\u200a1.75 [1.19\u20132.59]). Divergent effects were observed for tumor Gleason score, with two genes being associated with a higher Gleason score and two genes with a Gleason score <7 .To address the first of these, the expression levels of QTL candidate genes were examined in two publicly-accessible prostate cancer gene expression datasets using cBioPortal for Cancer Genomics ; and 2) a set of two genes with a protective effect (CCL19 and HIST1H1A). The expression levels of transcripts within these two gene sets were correlated with disease free survival (DFS) using Kaplan-Meyer survival analysis in the TCGA cohort. Specifically, DFS was compared between cases with higher or lower levels of expression of one or more gene in either of the two gene sets to those cases with normal levels of expression of the same genes. Eighteen percent (45/246) of cases in TCGA dataset exhibited divergent levels of one or more of the five genes positively correlated with aggressive disease development and GSE21032 cohorts were combined to create two gene sets: 1) a set of five genes associated with an increased propensity for aggressive disease development and GSE21032 are correlated with primary tumor copy number variation. Finally, the expression levels of the two genes that were negatively correlated with aggressive disease on logistic regression were not correlated with DFS in either cohort.Our QTL mapping strategy demonstrates that QTL candidate gene germline variation is associated with aggressive tumorigenesis in the TRAMP mouse. To evaluate whether this is the case for the human orthologs of these genes, SNP allele frequencies were evaluated in a publicly available human prostate cancer GWAS dataset. Specifically, these analyses were performed using the Cancer Genetic Markers of Susceptibility (CGEMS) GWAS, which consists of 1,172 prostate cancer patients and 1,157 controls of European ancestry from the Prostate, Lung, Colon and Ovarian (PLCO) Cancer Screening Trial Given that we hypothesized that QTL candidates modulate prostate cancer aggressiveness but not prostate cancer initiation, controls were omitted from analyses. The CGEMS cohort is well suited for this purpose, with the case cohort subdivided into non-aggressive and aggressive cases. In addition to these clinical characteristics, case-case analyses were performed for the additional aggressive disease variables shown in CXCL14, ITGAX, and LPCAT2) were all associated with aggressive prostate cancer: for CXCL14, associations were evident between rs801564 and metastasis to regional lymph nodes , and between rs10515473 and Gleason score at prostatectomy ; for ITGAX, an association was apparent between rs8047538 and Gleason score at prostatectomy ; and for LPCAT2, associations were evident between rs3764263 and primary tumor stage , between rs289707 and biopsy Gleason score , between rs2289119 and metastasis to regional lymph nodes , and between rs17369578 and best Gleason score available . Manhattan plots for all relevant genomic regions are shown in In the study, 1,317 SNPs were mapped within a 100 kb radius of the 29 QTL candidate genes were tested in the CGEMS cohort. Analysis of aggressive vs. non-aggressive disease phenotypes were performed as per the comparisons described in LPCAT2 are observed in a diverse range of tumors, notably breast and cervical carcinomas CXCL14 resides in a risk locus for aggressive prostate cancer in the 5q31 region CXCL14 in fibroblasts stimulates tumor angiogenesis and growth of prostate cancer cells CXCL14 in bulk tumor tissue is associated with an increased risk of recurrence.A systems genetics approach has been employed in this study to identify three novel susceptibility genes for aggressive prostate cancer, and to the best of our knowledge, this is the first study of its type to use this approach in this form of cancer. The three high priority candidate genes identified in QTL mapping studies using the TRAMP mouse model have diverse cellular functions , and havCxcl14/CXCL14 being negatively associated with primary tumor burden in (TRAMP \u00d7 NOD/ShiLtJ) F2 mice but positively correlated with disease recurrence in humans. Additionally, the traits used to nominate candidate genes in (TRAMP \u00d7 NOD/ShiLtJ) F2 mice frequently differ from the associated aggressive disease traits observed in human populations, as illustrated in Identification of these novel aggressive prostate cancer susceptibility genes has been facilitated through use of the TRAMP mouse model. However, the NE histological phenotype of tumors and the use of the non-physiological SV40 T-antigen to induce tumorigenesis have led to criticism of TRAMP P-values that are both more stable and accurate than uncorrected values. Second, we also recognize that a genome-wide level of significance was not achieved with any of the SNPs characterized in the CGEMS GWAS dataset. One probable reason for this is the limited statistical power of the case-case analysis performed here, which reflects the relatively small study population. Validation of these findings in additional prostate cancer cohorts is therefore vital. However, this lack of genome-wide significance may reflect one of the few limitations of GWAS. Specifically, although GWAS have revolutionized our understanding of complex trait susceptibility, they have not yet been able to explain the complete influence of heritability on disease susceptibility. This is true of prostate cancer, where all of the variants thus far identified by GWAS are estimated to explain less than one third of familial disease risk P<0.05 nominal level of significance are being missed since they do not reach the necessarily stringent level of genome-wide significance The integration of these different lines of evidence from human prostate cancer datasets to validate findings from our genetic screen in the TRAMP mouse has proven a pivotal element of this study. There are, however, a number of aspects of our analysis of the CGEMS GWAS data that warrant further discussion. First, we acknowledge that our use of a permutation test does not fully resolve the issue of correcting for type I errors. Rather, permutation testing has allowed us to report CXCL14, ITGAX and, LPCAT2 as novel susceptibility genes for aggressive prostate cancer development. This is the first study of its type to address the influence of germline polymorphism on tumor progression and metastasis in prostate cancer using systems genetics approach. Additionally, this approach has identified novel modifiers of aggressive prostate cancer that might not be readily apparent through human association studies. Knowledge of these variants will allow for more accurate determination of a patient's risk of metastasis, thus improving prognostic accuracy and facilitating more personalized treatments.In summary, we have identified C57BL/6J-Tg(TRAMP)824Ng/J (TRAMP) and NOD/ShiLtJ mice were obtained from The Jackson Laboratory . F1 mice were generated by crossing TRAMP females, which were hemizygous for PB-TAg transgene (Tg), to NOD/ShiLtJ males. F2 mice were generated by crossing Tg+ F1 females with Tg- F1 males. All animals were handled, housed and used in the experiments humanely in accordance with the NHGRI Animal Care and Use Committee guidelines. All work was performed under Animal Study Protocol G-09-2. Mouse tail genomic DNA was extracted from F1 progeny with the HotSHOT method As described previously in P<0.001.Genomic DNA was extracted from F2 tail biopsies using a Gentra Puregene DNA Extraction Kit , per the manufacturers protocol. Five microliters of DNA at 75 ng/\u00b5l was used for SNP genotyping using the 1536 plex assay kit and GoldenGate Assay Mouse Medium Density Linkage Array following the manufacturers protocol . The intensity data for each SNP for 228 samples were normalized and the genotypes assigned using Illumina GenomeStudio Genotyping Analysis Module version 1.9.4. SNPs with a GC score <0.7 and non-informative (homozygous) SNPs were excluded from further analysis. SNP Hardy\u2013Weinberg equilibrium (HWE) P-values were estimated with PLINK. SNPs were omitted if the HWE th percentile excluded. This encompassed the average experiment-wide background intensity of 3.04\u00b10.12.As described previously in Microarray data are available through Gene Expression Omnibus (accession no. GSE58829).QTL analysis was performed using J/qtl eQTL analysis was performed using Matrix-eQTL in R P-values were calculated for all traits other than those with a binary distribution by correlating the log2 transformed expression intensities of all probes mapped to a given QTL with the relevant QTL trait using MedCalc . For the latter, student's t-tests were performed to test the significance of transcript-trait correlations. Correction for multiple testing was performed using the Benjamini-Hochberg FDR method using the QVALUE module in R Pearson correlation coefficients and associated http://www.cbioportal.org; https://tcga-data.nci.nih.gov/tcga/tcgaCancerDetails.jsp?diseaseType=PRAD&diseaseName=Prostate%20adenocarcinoma); and b) GSE21032 - Prostate Oncogenome Project, Taylor et al. QTL candidate gene expression levels were analyzed in the cBioPortal for Cancer Genomics database (http://cbio.mskcc.org/cancergenomics/prostate/data/MSKCC_PCa_mRNA_data.zip). The expression levels of the 29 QTL candidate genes were subsequently extracted of all primary tumors with mRNA data (n\u200a=\u200a131), average expression levels and standard deviations calculated, and z-scores for candidate gene expression in individual tumors calculated using the following formula: .In the GSE21032 cohort, gene up- or down-regulation in a given case is again provided by cBioPortal as a z-score of \u22652 or \u2264\u22122, respectively. However, here a z-score of 2 was defined as an array probe-set intensity that is two standard deviations greater than the mean of the probe set intensity in the matched normal tissue, with the opposite being true for down-regulated genes. Therefore, to make candidate gene expression levels more comparable to those reported for TCGA cohort, raw gene expression data for GSE21032 were downloaded from cBioPortal . Logistic regression was performed using the stepwise method, with individual dichotomized clinical variables as depenP-values were estimated with PLINK. SNPs were omitted if the HWE P<0.001. Association analysis between aggressive prostate cancer phenotype and SNP or haplotype was performed using a generalized linear model (glm). Age and PC1, PC2 and PC3 were included as covariates in the glm. Analysis of aggressive vs. non-aggressive disease phenotypes were performed as per the comparisons described in http://biowulf.nih.gov). Specifically, permutation testing was performed for each phenotype against one SNP under rearrangements of the labels on all individuals with 10,000 times. Permutation tests were performed only in instances where the uncorrected P<0.01. Manhattan plots were constructed in R. For haplotype analysis, genome-wide LD blocks were estimated by using the Solid Spine algorithm of Haploview software with the default parameters, and fastPHASE was performed to generate haplotypes for each individual based on the LD blocks on NIH biowulf super cluster computer system (http://biowulf.nih.gov). FDR P-values were calculated by the MULTITEST package of R. All analyses were performed by using R.The clinical characteristics of the CGEMS GWAS cohort have been described extensively elsewhere F2 mice. Primary prostate tumor burden exhibited a negative correlation with age of death (A) and positive correlations with DMFS (B), lymph node metastasis (C), and lymph node metastasis burden (D). Conversely, seminal vesicle tumor burden was positively correlated with age of death (E) and negatively correlated with DMFS (F), lymph node metastasis (G), and lymph node metastasis burden (H).(TIF)Click here for additional data file.Figure S2Correlation between seminal vesicle tumor burden and primary tumor burden in (TRAMP \u00d7 NOD/ShiLtJ) F2 mice.(TIF)Click here for additional data file.Figure S3QTL plots for aggressive disease loci identified in (TRAMP \u00d7 NOD/ShiLtJ) F2 mice. QTLs were observed for the following traits: (A) DMFS; (B) total nodal metastasis burden; (C) liver surface metastasis count; (D) prostate tumor burden; (E) seminal vesicle tumor burden; and (F) age of death. The horizontal dotted line represents a genome-wide level of statistical significance of \u03b1 <0.05.(TIF)Click here for additional data file.Figure S4Clinical characteristics of patients represented in the GSE21032 and TCGA datasets.(TIF)Click here for additional data file.Figure S5Manhattan plots for genomic regions of interest in CGEMS GWAS. Plots are only shown for regions where candidate gene SNPs were associated with the following phenotypes: (A) best Gleason score available; (B) biopsy Gleason score; (C) prostatectomy Gleason score; (D) prostate cancer stage; (E) distant metastasis; (F) nodal involvement; and (G) primary tumor stage.(TIF)Click here for additional data file.Table S1Distribution of aggressive prostate cancer phenotypes across (TRAMP \u00d7 NOD/ShiLtJ) F2 mice.(XLSX)Click here for additional data file.Table S2Proximal eQTLs in (TRAMP \u00d7 NOD/ShiLtJ) F2 mice.(XLSX)Click here for additional data file.Table S3trans-eQTLs in (TRAMP \u00d7 NOD/ShiLtJ) F2 mice.Distal and (XLSX)Click here for additional data file.Table S4Correlation analysis for the microarray expression level of transcripts located within the chromosome 1 DMFS QTL with DMFS.(XLSX)Click here for additional data file.Table S5Correlation analysis for the microarray expression level of transcripts located within the chromosome 11 DMFS QTL with DMFS.(XLSX)Click here for additional data file.Table S6Correlation analysis for the microarray expression level of transcripts located within the chromosome 13 lymph node metastasis burden QTL with lymph node metastasis burden.(XLSX)Click here for additional data file.Table S7Correlation analysis for the microarray expression level of transcripts located within the chromosome 11 liver surface metastasis count QTL with liver surface metastasis count.(XLSX)Click here for additional data file.Table S8Correlation analysis for the microarray expression level of transcripts located within the chromosome 13 primary tumor burden QTL with primary tumor burden.(XLSX)Click here for additional data file.Table S9Correlation analysis for the microarray expression level of transcripts located within the chromosome 2 seminal vesicle tumor burden QTL with seminal vesicle tumor burden.(XLSX)Click here for additional data file.Table S10Correlation analysis for the microarray expression level of transcripts located within the chromosome 4 seminal vesicle tumor burden QTL with seminal vesicle tumor burden.(XLSX)Click here for additional data file.Table S11Correlation analysis for the microarray expression level of transcripts located within the chromosome 8 seminal vesicle tumor burden QTL with seminal vesicle tumor burden.(XLSX)Click here for additional data file.Table S12Correlation analysis for the microarray expression level of transcripts located within the chromosome 17 seminal vesicle tumor burden QTL with seminal vesicle tumor burden.(XLSX)Click here for additional data file.Table S13Correlation analysis for the microarray expression level of transcripts located within the chromosome 7 age of death QTL with age of death.(XLSX)Click here for additional data file.Table S14Correlation analysis for the microarray expression level of transcripts located within the chromosome 8 age of death QTL with age of death.(XLSX)Click here for additional data file.Table S15Statistically significant aggressive disease-associated haplotypes for QTL candidate genes in the CGEMS prostate cancer cohort.(XLSX)Click here for additional data file."} +{"text": "Inflammatory neuropathies represent disabling human autoimmune disorders with considerable disease variability. Animal models provide insights into defined aspects of their disease pathogenesis. Forkhead box P3 (FoxP3)+ regulatory T lymphocytes (Treg) are anti-inflammatory cells that maintain immune tolerance and counteract tissue damage in a variety of immune-mediated disorders. Dysfunction or a reduced frequency of Tregs have been associated with different human autoimmune disorders. We here analyzed the functional relevance of Tregs in determining disease manifestation and severity in murine models of autoimmune neuropathies. We took advantage of the DEREG mouse system allowing depletion of Treg with high specificity as well as anti-CD25 directed antibodies to deplete Tregs in mice in actively induced experimental autoimmune neuritis (EAN). Furthermore antibody-depletion was performed in an adoptive transfer model of chronic neuritis. Early Treg depletion increased clinical EAN severity both in active and adoptive transfer chronic neuritis. This was accompanied by increased proliferation of myelin specific T cells and histological signs of peripheral nerve inflammation. Late stage Treg depletion after initial disease manifestation however did not exacerbate inflammatory neuropathy symptoms further. We conclude that Tregs determine disease severity in experimental autoimmune neuropathies during the initial priming phase, but have no major disease modifying function after disease manifestation. Potential future therapeutic approaches targeting Tregs should thus be performed early in inflammatory neuropathies. Inflammatory polyneuropathies constitute disabling disorders of the peripheral nervous system (PNS) including acute and chronic variants. The acute Guillain-Barr\u00e9 syndrome (GBS) features rapid onset, monophasic PNS inflammation Regulatory T cells (Tregs) expressing the transcription factor forkhead box protein 3 (FoxP3) are a naturally occuring anti-inflammatory T cell subset that is indispensable for the maintenance of self tolerance and immune homeostasis 180\u2013199) (JPT peptide technologies) emulsified in 100 \u00b5l complete Freund's adjuvant (CFA) (Difco) containing 1 mg/ml heat inactivated Mycobacterium tuberculosis strain H37RA mixed with 100 \u00b5l PBS into the flanks. Animals received intraperitoneal injections of 500 ng pertussis toxin (PTx) (Sigma-Aldrich) dissolved in 100 \u00b5l sterile phosphate buffered saline (PBS) on the day of immunization (day 0) and on day 2 after immunization (day 2). To achieve Treg depletion, DTx (Merck) was dissolved in 100 \u00b5l sterile PBS and applied intraperitoneally on d3 and d4 at 50 \u00b5g/kg (1 \u00b5g per mouse). Two independent experiments were performed including 4\u20136 animals per group respectively. A modified clinical EAN score Generation of mice expressing the human diphtheria toxin (DTx) receptor under the control of the FoxP3 promotor (DEREG mice) has been previously described Animals were maintained at the central animal facility of the Heinrich-Heine-University, D\u00fcsseldorf under specific pathogen free conditions and transferred to conventional housing for experimentation. Animal experimentation was approved by the responsible state authorities with the approval reference numbers 8.87\u201350.10.34.08.207 and 50.05-230-65/06. Active EAN was induced by subcutaneous injection of 2,5 mg lyophilized bovine peripheral nerve myelin (bPNM) as previously described -/- NOD mice spontaneously develop a chronic inflammatory neuropathy that can be adoptively transferred to immunodeficient hosts by isolated CD4+ T cells -/- NOD mice. After 48h in culture, cells from all donor animals were pooled, washed three times with PBS, adjusted to 23\u00d7106 cells suspended in 400 \u00b5l PBS per animal and intravenously injected into 6 to 8 weeks old host immunodeficient severe combined immunodeficient (SCID) mice on NOD background (Taconic) subsequently named NOD-SCID mice. Recipient mice were maintained under specific pathogen free conditions for up to 10 weeks and were analyzed for phenotypic signs of neuropathy twice per week by a blinded investigator (S.C.). Treg depletion was performed by intraperitoneal antibody application as described above on days 6 to 10 (early depletion) and days 37 to 40 (late depletion) after adoptive transfer. Electrophysiogy was performed at the end of the experiment 50 and 70 days after transfer in early and late depleted animals, respectively.ICAM-1The standardized grip strength test was performed for hind limbs as previously described Mouse sciatic nerve conduction properties were determined as previously described 2 section area was calculated.Animals were sacrificed by cervical dislocation and directly intracardially perfused with phosphate buffered saline (PBS) followed by 4% paraformaldehyde. Sciatic nerves were dissected, paraffin embedded, cut into 7 \u00b5m sections on a standard microtome and haematoxylin-eosin (HE) stained following standard protocols. For immunohistochemical staining, nerve sections were incubated with anti-mouse CD3 antibody followed by a biotinylated goat anti-rabbit secondary antibody and a streptavidin-biotin-horseradish peroxidase complex (DAKO). 3,3\u2032-diaminobenzidine (DAB) was added as peroxidase substrate according to manufacturer's instructions. Between all protocol steps sections were washed for five minutes in PBS. Slides were dehydrated and mounted in xylene based medium (Merck). All incubations were performed at room temperature. The entire sciatic nerve section was photographed using a standard microscope (Zeiss) and photographs were photomerged using Photohop CS3 (Adobe). The total endoneural area was measured and the number of CD3 reactive cell nuclei was counted on merged photographs of the sections by a blinded investigator (S.C.) using the CellCounter plug-in of ImageJ . The density of CD3+ cells per mm7/well) in 6-well plates stimulated using soluble antibodies against CD3 and CD28 at 37\u00b0C in a humidified CO2 incubator for adoptive transfer, proliferation experiments or stained for flow cytometry. Blood taken by tail vein bleeding was collected in 75 \u00b5l hematocrit capillaries and transferred into 1 ml of PBS/1%FCS/10 mM EDTA solution followed by erythrocyte lysis. Flow cytometry was performed from blood and spleen cells. Cells were stained for cell surface CD4 , CD25 and intracellular FoxP3 using the FoxP3 staining buffer set . Flow cytometry was performed using a FACSCanto II flow cytometer (BD Biosciences). To assess the autoimmune proliferatory response after Treg depletion, splenocytes from NOD-SCID mice having previously received adoptive transfer of ICAM-1-/- NOD lymphocytes and injections of anti-CD25 antibodies were cultured in the presence of syngenic mouse sciatic nerve homogenisate. 2\u00d7105 splenocytes were maintained in 96-well plates for 96 hours. 3H-Thymidin was added for the last 24 hours and proliferation was assessed in quadruplicate wells by measuring 3H-Thymidine incorporation. Stimulatory indices were calculated by dividing counts per minute (CPM) of each well by the average CPM of non-stimulated wells.Splenocytes were extracted from dissected spleens by passing through a 40 \u00b5m cell strainer followed by erythrocyte lysis (both BD Biosciences). Splenocytes were either cultured . Data were statistically analyzed using GraphPadPrism 5.0 (GraphPad Software). The Wilcoxon-Mann-Whitney and Student's t-test for unrelated samples were used to test for statistically significant differences of non-Gaussian and Gaussian distributed data, respectively. Differences were considered significant at p-values <0.05.We used different approaches to study the importance of Tregs in EAN. The previously described DEREG mouse line expresses the human DTx receptor under the control of the FoxP3 promotor together with a GFP reporter We therefore utilized anti-CD25 directed antibody mediated Treg depletion as previously described in models of central nervous system inflammatory demyelination -/-) NOD mice is experimentally difficult to utilize due to its highly variable onset -/- NOD mice into immunodeficient hosts thus triggering neuritis with synchronized onset but at the same time clinically resembling aspects of human chronic inflammatory neuropathies. Again application of the CD25 antibody reduced the percentage of FoxP3+ cells in NOD-SCID recipients. This reduction exhibited a non-significant trend in early depleted animals analyzed 40 days after depletion (Murine EAN in our hands features low disease severity, making clinical assessment difficult in this model. Also, the spontaneous neuritis in ICAM-1 deficient (ICAM-1epletion and was epletion .-/- NOD lymphocytes into NOD-SCID mice caused progressive impairments first manifesting 20 days after transfer (-/- NOD mice show autoreactivity against components of their peripheral nerve myelin Adoptive transfer of ICAM-1transfer . Early Ttransfer . Animal transfer . Electrotransfer and avertransfer . We have+ Tregs during acute stages of GBS while their suppressive function was unaltered +CD25+ T cells, which may include activated T effector cells in the acute phase of GBS without studying FoxP3 expression + Tregs Tregs are indispensable for the maintenance of self tolerance and immune homeostasis and Treg dysfunction has been associated with various human autoimmune disorders and has been discussed as one potential factor driving disease progression. Reduced levels or an impaired function of Tregs have been described in human patients with acute and chronic inflammatory neuropathies. One study reported a transiently reduced number of FoxP3Previous studies have not addressed, however, if Tregs functionally determine the manifestation of inflammatory neuropathies and if Treg numbers may represent a factor determining chronicity of autoimmunity in the peripheral nerve. We here demonstrate that depletion of Tregs increases the disease severity of actively induced and adoptively transferred murine autoimmune neuritis models. We conclude that Tregs suppress autoinflammatory reactions in the PNS and determine disease severity in inflammatory neuropathies. In synopsis with the previously published descriptive studies in human neuritis patients and corresponding animal models our findings are thus the first to demonstrate a functional relevance of Tregs in peripheral neuritis.Depletion had more pronounced effects if performed early in the course of the respective disease model, indicating that Tregs constrain inflammation during the priming phase of EAN and during the early phases of adoptively transferred neuritis. Although Tregs are required to prevent autoimmunity throughout the lifespan of mice Our analysis of Tregs was impeded by several methodical difficulties. Firstly, when establishing the DEREG mouse model, we found that DTx could not be combined with active immunization of any kind due to a non-cell type specific probably septic reaction Initially EAN was established in different non-murine species and preferentially used in the Lewis rat strain -/- NOD mice We additionally used an adoptive transfer paradigm to elicit a severe progressive autoimmune neuropathy previously described in ICAM-1In conclusion, we have demonstrated the functional relevance of Tregs for the manifestation and severity of inflammatory neuropathies. In synopsis with previous descriptive human data, this identifies Tregs as potential therapeutic target in the early stage of disabling inflammatory neuropathies."} +{"text": "Current axillary lymph node assessment criteria by ultrasonography (US) depends on cortical thickness >3 mm, focal eccentric cortical thickness and lobulated cortex. Combination with a prediction tool may define better patient management strategies.Newly diagnosed breast cancer patients within a 1-year period were randomly selected retrospectively. Inclusion criteria include patients with final histological correlation following resection. The Memorial Sloan Kettering Cancer Centre breast cancer normogram has been utilised. Axillary nodal US findings and histological correlation from fine needle aspiration cytology (FNAC)/core biopsies were obtained.A total of 70/160 randomly selected patients fulfilled the inclusion criteria.In total, 28/70 patients were positive for axillary nodal metastases. A total 15/28 patients underwent FNAC/core biopsy and 11 were positive. In the remaining 13 patients, nodal metastases were detected by sentinel lymph node biopsy (SLNB) in 10 patients and initial axillary node clearance (ANC) in three patients. These 13 patients had a probability range of 26 to 91% (mean 59%). The overall probability range in all 28 patients is 6 to 97% (mean 64%). Forty-two patients did not have lymph node metastases. One patient did not have axillary US assessment. In total, 10/41 patients had abnormal nodes on US that were negative on FNAC/core biopsy. All 42 patients had surgical lymph node assessment . The overall probability range is 9 to 80% (mean 36%).The detection of axillary nodal metastases with US remains low but combination with the normogram prediction tool may be helpful to determine patients with high probability to have repeated US assessment and sampling of normal-looking nodes to increase the detection rate."} +{"text": "It represents the most frequent genetic cause of end-stage renal disease in the first three decades of life. NPH is characterized by the dysfunction of sensory cilia which explains the complexity of the NPH phenotype. It can be associated with retinitis pigmentosa (Senior-L\u00f8ken syndrome), mental retardation and ataxia (Joubert syndrome), skeletal anomalies (Jeune syndrome), or To date, recessive mutations causing NPH have been identified in more than eighteen different genes (NPHP1-NPHP18). Their gene products localize at the primary cilia-centrosome complex, along the cilium as intraflagellar transport proteins and are important in signaling pathways downstream of cilia including Wnt signaling, Shh signaling and the DNA damaged response pathway.TRAF3IP1 in patients presenting with NPH, retinitis pigmentosa, skeletal defects of the pelvis, hexadactyly and hepatic fibrosis. TRAF3IP1 encodes IFT54 which is involved in the anterograde transport along the primary cilia.Using whole and targeted exome sequencing, we identified novel protein altering mutations in TRAF3IP1 knock-down cells confirmed the observed defects in microtubule organization. Furthermore, sphere formation assays as well as the pronephros of elipsa zebrafish embryos showed defects in epithelialization.Besides its known function in cilia we demonstrate that TRAF3IP1 act as a key regulator of cytoplasmic microtubule organization. Mass spectrometry analyses as well as pull-down experiments demonstrated that mutations in TRAF3IP1 lead to an altered binding to actin and microtubule associated proteins. Immunofluorescence stainings using patient fibroblasts as wells as mIMCD3 TRAF3IP1 affect both ciliary and non-ciliary functions of TRAF3IP1 which can provide an explanation for kidney tubules morphogenesis defects as well as the other disease phenotypes e.g. retinal, skeletal and hepatic defects.Altogether our findings demonstrate that NPH causing mutations of"} +{"text": "Escherichia coli (APEC) is associated with colibacillosis in poultry. Here, we present the first complete sequence of an APEC strain of the O7:HNT serotype and ST73 sequence type, isolated from a broiler with cellulitis. Complete genomes of APEC with distinct genetic backgrounds may be useful for comparative analysis.Avian pathogenic Escherichia coli is a versatile bacterium exhibiting a high degree of genomic plasticity or extraintestinal diseases in humans and animals (\u2013E.\u00a0coli (APEC) . Collecti (APEC) . Colibaci (APEC) . Besidesi (APEC) .E.\u00a0coli (UPEC) IAI39 (accession no. CU928164), UPEC UMN026 (E.\u00a0coli (NMEC) CE10 10), and C UMN026 , and neoEC) CE10 . To our E.\u00a0coli strain RS76 of the serotype O7:HNT and sequence type ST93 was isolated from the carcass of a slaughtered broiler diagnosed with avian cellulitis in Brazil (De novo assembly was performed with SPAdes 3.0 (E.\u00a0coli BL21(DE3) (accession no. AM946981) as a reference. Gaps were eliminated with PCR and subsequent Sanger sequencing. The APEC RS76 genome sequence was annotated by the NCBI Prokaryotic Genomes Automatic Annotation Pipeline (PGAAP). The consistency of the PGAAP annotation was verified against a previous Prokka 1.11 annotation . This chromosome presented 4,407 coding sequences, 84 tRNA encoding genes, and 7 rRNA encoding operons.There are two general issues regarding the impact of APEC that still need to be elucidated. The genetic determinants associated with APEC pathogenicity in avian species are not fully understood yet. Also, the role of APEC as a zoonotic agent is controversial. The sequencing of complete APEC genomes presenting distinct genetic backgrounds may be useful for future comparative genomic analysis addressing those issues.CP013048.The complete sequence of strain RS76 was deposited in GenBank under the accession number"} +{"text": "APOBEC3G and Vif genes. This kind of conflict leads to rapid fixation of mutations that alter amino acids at the protein\u2013protein interface, referred to as positive selection. We show that the APOBEC3G gene has been subject to strong positive selection throughout the history of primate evolution. Unexpectedly, this selection appears more ancient than, and is likely only partially caused by, modern lentiviruses. Furthermore, five additional APOBEC genes in the human genome appear to be engaged in similar genetic conflicts, displaying some of the highest signals for positive selection in the human genome. Despite being only recently discovered, editing of RNA and DNA may thus represent an ancient form of host defense in primate genomes.Host genomes have adopted several strategies to curb the proliferation of transposable elements and viruses. A recently discovered novel primate defense against retroviral infection involves a single-stranded DNA-editing enzyme, APOBEC3G, that causes hypermutation of HIV. The HIV-encoded virion infectivity factor (Vif) protein targets APOBEC3G for destruction, setting up a genetic conflict between the APOBEC3G, a gene that edits retroviral DNA like HIV, is under positive selection that predates the origin of HIV, implying that RNA/DNA editing represents an ancient form of intragenomic host defense Neurospora crassa being a notable exception 117\u2013250).The discovery that inst HIV . An unbi/Ks test , which a(APOBEC1 and APOBEC3C), and there was insufficient chimpanzee sequence available in one case (APOBEC2). To gain further information about these genes, we sequenced them from either orangutan or gorilla . Although we might have expected APOBEC1 to be evolving only under purifying selection based on its important editing of APOB mRNA, our analysis suggests that APOBEC1 has also participated in some kind of genetic conflict involving its first active site, and suggests that the rapid evolution of APOBEC1 seen previously in mouse\u2013rat comparisons may also be due to positive selection ((APOBEC2) and positive (APOBEC3E) selection. These findings greatly extend the current understanding of the APOBEC family, and implicate a majority of APOBEC genes as participants in host defense. They also raise the possibility of other editing systems being involved in genome defense; for instance, hepatitis delta virus is known to be edited by adenosine deaminase , especially in populations with a high incidence of HIV infection, since different alleles of APOBEC3G may have different susceptibility to various viral strains. The action of APOBEC3G on viral evolution could also be complex because, although it is ineffective as an antiviral mechanism in the presence of Vif, its action could also result in an increased likelihood of adaptive changes and viral diversity in the host due to the introduced G-to-A hypermutations. Polymorphisms in APOBEC3G may thus have direct impact on the progression time from initial HIV infection to AIDS, and should be investigated as such.The antiviral activity of APOBEC3G and the excess of non-synonymous changes specific to human EC3G see implicatAPOBEC3G does not identify the biological step that exerts this selective pressure. Formally, this step could be the yet-undefined mechanism by which APOBEC3G is packaged into virions, the interaction of APOBEC3G with Vif-like destruction proteins encoded by other viruses, and/or its interaction with the proteasome machinery. APOBEC3G may indeed interact with other viruses, because G-to-A hypermutation\u2014a hallmark of the single-stranded DNA\u2013editing activity of APOBEC3G-like enzymes\u2014has been observed in some nonlentivirus viruses (NAO3448A), Pan paniscus (bonobo) (NGO5253), Gorilla gorilla (gorilla) (NG05251B), Pongo pygmaeus (orangutan) (NAO4272), Macaca nigra (Celebes crested macaque) (NG07101), Macaca fascicularis (crab-eating macaque) (NA03446), Erythrocebus patas (patas monkey) (NG06254), Lagothrix lagotricha (common woolly monkey) (NG05356), and Saguinus labiatus (red-chested mustached tamarin) (NG05308). Papio anubis (baboon) DNA was a personal gift from Dr. Trent Colbert. The APOBEC3G, APOBEC1, APOBEC2, and APOBEC3C genes were amplified exon-by-exon from genomic DNA with PCR Supermix High Fidelity , and PCR products were sequenced directly. PCR and sequencing primers are shown in APOBEC3G sequence was obtained from the Ensembl database of the human genome project (ENSG00000100289). The Chlorocebus aethiops (African green monkey) APOBEC3G sequence (GenBank AY331714.1) is missing the last 21 bp of the coding sequence because it was sequenced from mRNA (APOBEC3G from NWMs (woolly monkey and tamarin) where the \u201cAG\u201d directly 5\u2032 of the eighth coding exon is missing. Sequences have been deposited in GenBank under the following accession numbers: APOBEC3G (AY622514\u2013AY622593), APOBEC3C (AY622594\u2013AY622597), APOBEC2 (AY622598\u2013AY622599), APOBEC1 (AY622600\u2013AY622604).Genomic DNA was obtained from Coriell . Species and Coriell repository numbers are: rom mRNA in a preAPOBEC1 (ENSG00000111701), APOBEC2 (ENSG00000124701), AID (ENSG00000111732), APOBEC3A (ENSG00000128383), APOBEC3B (NM_004900.3), APOBEC3C (ENSG00000179750), APOBEC3DE (ENSG00000179007), and APOBEC3F (ENSG00000128394). Transcripts for both APOBEC3D (NM_152426) and APOBEC3DE (BC017022.1) exist in the database. Chimp sequences were obtained from orthology to human genes assigned on the University of California at Santa Cruz Genome Bioinformatics Website (http://www.genome.ucsc.edu). All orthologous chimp exons were checked for AG and GT flanking the 5\u2032 and 3\u2032 boundaries, respectively, an indication that human splice sites are conserved. The mouse APOBEC3 protein sequence can be found in GenBank (NP_084531.1).Human sequences were obtained from the Ensembl or GenBank databases: DNA sequences were aligned using Clustal_X , with hap < 10\u221213) with a significant fraction of the sites (more than 30%) predicted to evolve at average \u03c9 ratios greater than 3.5 has undergone adaptive evolution in at least three distinct periods. Despite being only 29 codons long, this domain has accumulated ten non-synonymous changes and only two synonymous changes in the African green monkey since it and the patas monkey last shared a common ancestor. Similarly, the orangutan has retained eight non-synonymous changes and no synonymous changes since it split from the rest of the hominids. Finally, a ratio of 6:0 R:S changes is seen in the split between the NWMs and the common ancestor of OWMs and hominids. Surprisingly, even the two active site structures of APOBEC3G (B and E) show evidence for adaptive evolution , including along the branch leading to the common ancestor of all hominids. The first pseudoactive domain (D) acquired ten non-synonymous and no synonymous changes since the hominids split from the OWMs.(343 KB PDF).Click here for additional data file.Figure S2p <10\u221213) (C) with a significant fraction of the sites (more than 30%) predicted to evolve at average \u03c9 ratios greater than 3.5. These analyses also identified certain amino acid residues with high posterior probabilities (greater than 0.95) of having evolved under positive selection (A and B).Maximum likelihood analysis was performed on APOBEC3G sequences using the PAML software package. To detect selection, the multiple alignments were fitted to either the F3\u00d74 (A) or F61 (B) models of codon frequencies. We compared the log-likelihood ratios of the data using comparisons of different NSsites models: model 1 versus model 2 and model 7 versus model 8 . In both cases, permitting sites to evolve under positive selection gave a much better fit to the data ((60KB PDF).Click here for additional data file.Figure S3The individual domains of the APOBEC3G protein are demarcated. Catalytically important residues are highlighted in bold, and those residues identified by PAML analysis as being under positive selection are indicated with gray shading. Blue shading highlights the single amino acid residue that can switch specificity of Vif interaction with APOBEC3G. AGM, African green monkey.(46 KB PDF).Click here for additional data file.Table S1(39 KB PDF).Click here for additional data file.http://www.ncbi.nlm.nih.gov/) and Ensembl (http://www.ensembl.org/) accession numbers for the genes and gene products discussed in this paper are as follows. GenBank: APOBEC1 (AY622600\u2013AY622604), APOBEC2 (AY622598\u2013AY622599), mouse APOBEC3 (NP_084531.1), human APOBEC3B (NM_004900.3), APOBEC3C (AY622594\u2013AY622597), APOBEC3D (NM_152426), APOBEC3DE (BC017022.1), APOBEC3G (AY622514\u2013AY622593), and African green monkey APOBEC3G (AY331714.1). Ensembl : APOBEC1 (ENSG00000111701), APOBEC2 (ENSG00000124701), AID (ENSG00000111732), APOBEC3A (ENSG00000128383), APOBEC3C (ENSG00000179750), APOBEC3DE (ENSG00000179007), APOBEC3F (ENSG00000128394), and APOBEC3G (ENSG00000100289).The GenBank (http://www.coriell.undmj.edu/) repository numbers for primate genomic DNAs are Pan troglodytes (chimpanzee) (NAO3448A), Pan paniscus (bonobo) (NGO5253), Gorilla gorilla (gorilla) (NG05251B), Pongo pygmaeus (orangutan) (NAO4272), Macaca nigra (Celebes crested macaque) (NG07101), Macaca fascicularis (long-tailed macaque) (NA03446), Erythrocebus patas (patas monkey) (NG06254), Lagothrix lagotricha (common woolly monkey) (NG05356), and Saguinus labiatus (red-chested mustached tamarin) (NG05308).Coriell ("} +{"text": "Defensins are important components of innate immunity to combat bacterial and viral infections, and can even elicit antitumor responses. Clusters of defensin (DEF) genes are located in a 2 Mb range of the human chromosome 8p23.1. This DEF locus, however, represents one of the regions in the euchromatic part of the final human genome sequence which contains segmental duplications, and recalcitrant gaps indicating high structural dynamics.We find that inter- and intraindividual genetic variations within this locus prevent a correct automatic assembly of the human reference genome (NCBI Build 34) which currently even contains misassemblies. Manual clone-by-clone alignment and gene annotation as well as repeat and SNP/haplotype analyses result in an alternative alignment significantly improving the DEF locus representation. Our assembly better reflects the experimentally verified variability of DEF gene and DEF cluster copy numbers. It contains an additional DEF cluster which we propose to reside between two already known clusters. Furthermore, manual annotation revealed a novel DEF gene and several pseudogenes expanding the hitherto known DEF repertoire. Analyses of BAC and working draft sequences of the chimpanzee indicates that its DEF region is also complex as in humans and DEF genes and a cluster are multiplied. Comparative analysis of human and chimpanzee DEF genes identified differences affecting the protein structure. Whether this might contribute to differences in disease susceptibility between man and ape remains to be solved. For the determination of individual DEF gene repertoires we provide a molecular approach based on DEF haplotypes.Complexity and variability seem to be essential genomic features of the human DEF locus at 8p23.1 and provides an ongoing challenge for the best possible representation in the human reference sequence. Dissection of paralogous sequence variations, duplicon SNPs ans multisite variations as well as haplotypes by sequencing based methods is the way for future studies of interindividual DEF locus variability and its disease association. One obvious reason for these gaps is that the appropriate regions are enriched in sequences that are not tolerated by the cloning systems. The second possibility is that even if clones are available and amenable for sequencing, their sequences cannot be unambiguously aligned due to gap flanking segmental duplications. Generally, those duplicons are defined by >90% sequence identity and lengths of >1 kb and about 87% of all human ones are longer than 50 kb . The coding sequences of the gene copies differ and in some cases the reading frame contains premature termination codons. Blat search of the type III repeat subunit revealed three paralogs in the human genome. Two of these are located on chromosome 8p23.1 at about 12 Mb (flanked by repeats of type II), the third at 12p13.31. The paralogs are slightly rearranged in comparison to the subunits of III.1-5.Located adjacent to DEF clusters b .We manually inspected seven clones covering Pan troglodytes, ptr) whole genome shotgun (WGS) working draft , but trace data inspection indicates the presence of several different haplotypes. Additionally, ptrDEFB108 and ptrDEFB109p are not covered by any chimpanzee WD sequences. As an alternative to the WD approach, we sequenced for ptrDEF cluster b three BAC clones. Examination of SNPs in overlapping regions (104 kb) of the three clones [GenBank:AC150655], [GenBank:AC150656], [GenBank:AC150657] revealed three different haplotypes originating from one chimpanzee. The detected aa changes in human and chimpanzee defensins are illustrated in ptrDEFB108 is a pseudogene, since also the start codon ATG is changed into GTG.In order to compare the human chromosome 8p23.1 DEF region to the orthologous locus in our closest relative, we both employed the chimpanzee to 1:7 (proband 1). Interestingly, this haplotype is also found in the trace archives of chimpanzee and baboon. Furthermore, ratios of the individual haplotypes of DEFB104 as well as of DEFB4 indicate different DEF cluster b numbers in the four individuals. While proband 3 bears five copies, proband 4 most probably harbors eight copies or multiples thereof.In order to determine individual DEF copy numbers we PCR-amplified a 500 bp fragment of ls Table . Three ode facto gap, it better reflects the region in the sense of a \u201ehuman genome reference\u201c, since the clones harboring copies of DEF clusters b derive from three libraries and may therefore represent up to five alleles (library RP13 is represented by only one clone). Our assembly also reflects better the diversity of all available sequence data of this chromosomal region: 27 out of 32 finished clones are incorporated into the tiling path. The remaining five clones cannot be included in the assembly according to our quality criteria and therefore must be regarded as parts of additional copies or alleles. Furthermore we point out that the identification of a third copy of the DEF cluster b in the 360-kb-contig does not represent an allele of clusters b1 or b2 derived from an alternative library / donor, since besides four SCb clones one RP11 clone is incorporated. With respect to the RP11 library of which most of the DEF cluster b covering clones derive we conclude that at present sequence information of at least five variants of the cluster is available from a single individual: cluster b1 ; b2 (clones 16 \u2013 20); b3 (clone 30); b4 (clone 11) and b5 (clone 12). All these results are in agreement with the reported interindividual variability of DEF cluster b genes ; green/yellow signals) and CTB-415D8 [GenBank:AF228730]; red signals) visualized by FISH on metaphase chromosomes according to standard protocols ,53. MetaClick here for fileResolving LCR type II and III \"pairs\" on chromosomes at approx. 900 band stage. Probe CTB-415D8 [GenBank:AF228730]; LCR type II and III) generates two clearly separated FISH signals at 4p16 and 8p23, respectively : In contrast, probe SCb-561b17 [GenBank:AF238378]; LCR type I) yield a single signal at 8p23, solely , that is co-localized with the telomeric signal of probe CTB-415D8 [GenBank:AF228730]. Signals with lower intensity are indeterminable in this picture.Click here for fileptrDEF cluster a) are deduced from the chimpanzee WD and might therefore include sequencing errors. Those for ptrDEFB1, ptrDEFB107, ptrDEFB105, ptrDEFB103, ptrDEFB4 and ptrDEFB108 (DEF cluster b) are derived from high quality BAC sequences and the appropriate traces were visually inspected. The gray shadow indicates the motif of six cystein residues (except for DEFB107 with only five cysteins).DEF aa sequences with highlighted residues (bold) different between human and chimpanzee. Boxes: human aa \u2013 human position \u2013 chimpanzee aa. All aa positions refer to the following human protein accessions: DEFA6 = [GenBank:NP_001917]; DEFA4 = [GenBank:NP_001916]; DEFA1 = [GenBank:NP_004075]; DEFA5 = [GenBank:NP_066290]; DEFB1 = [GenBank:NP_005209]; DEFB107 = [GenBank:AAM93909]; DEFB105 = [GenBank:NP_689463]; DEFB103 = [GenBank:NP_061131]; DEFB4 = [GenBank:NP_004933]; DEFB108 = [GenBank:AAN33116]. Aa for the chimpanzee orthologs ptrDEFA6, ptrDEFA4, ptrDEFA1, ptr novel defensin similar to DEFA4, ptrDEFA5 and ptrDEFB1 , individual loci may have incorporated variable numbers of additional b clusters in either orientation. The proposed duplicon consists of two inverted LCRs flanking a DEF cluster b (top/bottom). The orientation of any DEF cluster b can change either by inverted duplication/crossover (i) or homologous recombination within inverted LCRs . Moreover, the proposed genomic structure indicates that even in a 'minimal' DEF locus one or both DEF clusters may be deleted due to homologous recombination between direct LCR copies (\u0394). Sequence features of the most distal LCR as well as LCRs type II (rectangles) are shown. Click here for file"} +{"text": "Neurocognitive dysfunction is associated with important socio-professional consequences and diminished quality of life. In the Neurocognitive Remediation Clinic of the Centre Hospitalier Universitaire Brugmann , patients suffering from neurocognitive dysfunctions related to common mental disorders are considered for Neurocognitive Remediation Therapy (NCRT), combining personalized computerized cognitive training and strategy training, with group sessions of physical rehabilitation and cognitive behavior therapy.This cross-sectional study aims to assess the efficacy of a 12 week (1day/week) NCRT program organized within the day clinic.Patients who completed the NCRT between March 2018 and June 2019 were eligible to participate. Efficacy was assessed using the cognitive failure questionnaire (CFQ) and a 17-item questionnaire assessing daily functioning. Current scores on the CFQ were compared to the scores before and after NCRT. Additionally NCF was retrospectively assessed through the neuropsychological test results before and after NCRT.Of the eligible 38 patients, 27 consented to participate (18 women/9 men); median age was 52 years, range (29-61); median time since stop NCRT was 7 months, range (4-17). Twenty patients (80%) reported improvement in daily function. Subjective neurocognitive function improved significantly immediately after NCRT and remained stable at time of assessment . After NCRT at least 1 neuropsychological subtest normalized in 25 patients (96.15%). Divided attention, long-term visual memory and planning improved in respectively 80%, 75% and 75% of the patients.Our innovative integrative program improves neuropsychological performances and sustainably ameliorate subjective neurocognitive and daily function.No significant relationships."} +{"text": "The genome comprises one circular chromosome and one plasmid and lacks antimicrobial resistance genes.The complete genome of hydrogen peroxide (H For PacBio sequencing, gDNA (3\u2009\u03bcg) was sheared to 7 to 12\u2009kb using the Megaruptor 3 followed by a cleanup using the AMPure PB beads . The Sequel system equipped with SMRT cells 1 M v3 tray (PacBio) was used to sequence the libraries constructed using the SMRTbell express template prep kit 2.0 (PacBio), generating 145,301 subreads with an plasmid . Adaptiv plasmid over 30 Limosilactobacillus fermentum IMDO130101 (GenBank accession no. NZ_LT906621) as the closest strain, with 99.47% sequence identity. Genome annotation using PGAP v6.2 .The biospecimens used in this study were provided by the Biobank of Apple Tree Dental Hospital after approval from the public institutional review board , BioProject (accession no. PRJNA852394), BioSample (accession no. SAMN29328592), and SRA (accession nos. SRX15900883 and SRX15900884).The 16S rRNA gene sequence, genome sequence, and the raw sequencing reads for DM072 were deposited in GenBank (accession nos."} +{"text": "FNDC3A, which influences different developmental processes in vertebrates, like Sertoli cell/spermatid adhesion in mice testis, bone traits in chicken, and fin development in zebrafish. To identify downstream molecular processes during vertebrate development we investigated gene expression profiles in the previously establishedfndc3azebrafish mutants via microarray analyses on 22 hpf embryos (26-somite stage). Our analyses imply distinct transcriptional profiles between genotype groups and hint to altered cell binding and catalytic activity infndc3amutants.The group of Fibronectin type III domain-containing protein super family splits into a large number of gene-orthologues and mediates a variety of cellular functions during development and disease. They act as anti-inflammatory factors, are linked to cell-cell-interactions, regulate cell signaling and are associated with different cancer types, like cervical and colorectal. One member of this gene family is FNDC3Aexpression in human odontoblasts . Functional experiments inSymplastic spermatids(sys) knockout mice indicated that Fndc3a is essential for cell adhesion between spermatids and Sertoli cells, resulting in sterile males after depletion . Our previous work has shown that interference with Fndc3a function in zebrafish (Danio rerio) CRISPR/Cas9 mutants results in defects during tail bud development and caudal fin regeneration . The purpose of this follow-up study was to investigate potential downstream targets of Fndc3a during zebrafish development.Fndc3a is one member of the FNDC-super family and contains nine fibronectin type III domains. These fibronectin type III domains are a common feature of a large number of extracellular proteins and are evolutionary conserved in a large number of species. Molecular studies have shownfndc3a CRISPR mutant line displays a 5 bp indel alteration in exon 13 , resulting in potential frameshift and a premature STOP codon. The corresponding molecular analyses clarified a hypomorphic and temperature-sensitive phenotype in the generatedfndc3aline, although homozygous embryos display a prominent tail fin phenotype during development and caudal fin regeneration defects . Earliest developmental changes in the ventral tail bud region of homozygous mutants was observed at 21-somite stage (19.5 hours post fertilization (hpf)).The establishedfndc3amutant line we performed Affymetrix microarray analyses. Whole-RNA was extracted from pools of 26-somite stage embryos (22 hpf) of different genotypes and was subsequently used for Gene Level Differential Expression Analysis. Each genetic condition was analyzed in biological triplicates and three independent embryo pools for each genotype were hybridized to Affymetrix Zebrafish Gene 1.0 ST Arrays. Unbiased post-run analyses showed clustering of genotypes and significant differences between genotype groups. Further detailed comparison between different genotype conditions indicated many up and down regulated transcripts . Besides a large number of unresolved or ill-described transcripts , a number of genes could be identified which have been previously linked to embryonic development in zebrafish (see suppl. table 1). These genes have been described to be essential for development of cells located in the tail bud region or show expression within the tail bud surrounding tissues at this stage and are thereby in accordance with the investigated developmental stage and the previously observedfndc3amutant phenotype. For example,anxa1awhich is involved in caudal fin regeneration shows a reduced expression in homozygous mutants , thus, matching with caudal fin regeneration defects observed in these mutants. In addition,gata1ais a transcription factor involved in hemopoiesis and is overexpressed in blood precursor cells at this stage . These cells are partly located in the blood island area, which is affected by ECM defects in ventral fin fold tissues offndc3amutants. A last example is overexpression ofvent , a homeobox gene which is embedded in a regulatory network along withvoxto repress dorsal cell fates during tail bud development in zebrafish . To identify generally affected molecular functions, biological processes and cellular components in the mutants we performed additional GO term analyses (PANTHER database). These analyses indicate that reduction or loss of Fndc3a function predominantly affects transcripts linked to binding and catalytic activity, and to cellular and metabolic processes . Future studies will have to elucidate functions of ill-described transcripts and will have to link Fndc3a function to well-described genes during early tail bud development in zebrafish.To further investigate early transcriptional and developmental alterations in the1) RNA extraction and microarray analyses:RNA extraction of three genetic genotype groups , each group consisting of 12 pooled embryos without chorion, has been performed with QIAzol lysis reagent according to RNeasy whole RNA extraction protocol (QIAGEN). Quality control of total RNA extraction via Eukaryote Total RNA Nano assay on a Bioanalyser (Agilent) and only RNA samples with RIN values larger than 5 were used for subsequent cDNA synthesis, fluorescent labeling and microarray hybridization. Quantification of transcriptional changes in mutants was done by Affymetrix microarray analyses using \u201cZebrafish Gene 1.0 ST Array\u201d and Affymetrix software packages (Affymetrix). Microarray experiments were performed in cooperation with Core Unit Systems Medicine W\u00fcrzburg according to Affymetrix specifications.2) Data analyses:Data analyses has been performed via Affymetrix Transcriptome Analysis Console software. GO term analyses were performed with PANTHER classification system version 10.0 (http://www.pantherdb.org/). Figure data and graphics have been assembled with Excel (Microsoft Corporation), OriginPro (OriginLab Corporation), and CorelDraw Graphics Suite (Corel Corporation). Original microarray data files are deposited in ArrayExpress and can be downloaded athttps://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-11958.3) Animal handling:All procedures involving experimental animals were performed in compliance with German animal welfare laws, guidelines, and policies. Generation of the usedfndc3a mutant strain and founder animal genotyping via fin clipping was approved by the Committee on the Ethics of Animal Experiments of the University of W\u00fcrzburg and the corresponding legislative body . The generatedfndc3amutant line has been previously described and is submitted to ZFIN.org . Zebrafish embryos used for this study were raised at 28.5\u00b0C in Danieau\u00b4s medium 2, 1.5 mM Hepes, pH 7.2, 0.001% phenol blue) and were staged according to Kimmel at al 1995.4) Statistical analysis: Statistical comparison of different genotype groups were performed with the following settings via Affymetrix Transcriptome Analysis Console software: One-Way Between-Subject ANOVA (unpaired); Fold Change (linear) < -2 or Fold Change (linear) > 2; ANOVA p-value (Condition pair) < 0.05; Triplicate conditions: wild type,fndc3ahet andfndc3ahomo. Gene Level Differential Expression Analysis with this settings indicated overall 405 differentially expressed genes from a total number of 59384 analyzed genes.10.22002/yn220-zt682Description: Extended excel table summarizing presented microarray data and analyses.. Resource Type: Dataset. DOI:"} +{"text": "This would help inform quality improvement interventions prior to adoption of the 2022 CQUIN, CCG2: Appropriate antibiotic prescribing for UTI in adults aged 16+.3UK CQUIN schemes encourage an improvement focus on a specific area of care. In 2019, CQUIN CCG1a: Improving the management of lower UTI in older people2Service evaluation of the UTI pathway including compliance with two of the CQUIN care processes for UTI diagnosis in patients age 65+ presenting to ED (not admitted): (i) diagnosis excludes use of urine dipstick in people aged 65+; and (ii) urine sample sent to microbiology as per UK guidance.2 presence/absence of urine dipstick test; and presence/absence of urine sample for culture and susceptibility testing. Findings were compared with identical trust data for patients (admitted and non-admitted) obtained during the 2019 CQUIN: Q1 April\u2013June; Q2 July\u2013September; Q3 October\u2013December. During this period improvement interventions were implemented.A search of the electronic patient record for key terms (Table 1) identified 6076 ED attendances for patients age 65+ between 1 August and 31 October 2021 of which 40 were identified with a primary diagnosis of UTI not requiring hospital admission. Paramedic, ED and Urgent Treatment Centre notes (paper and electronic) were reviewed in detail and information gathered regarding presence/absence of UTI symptoms aligned to diagnostic guidelines;See Table 2.3 consideration should be given to a bundle of interventions including education, data feedback and systems improvement, for example, computerized decision support systems (CDSS) to embed sustained change.Following intensive staff education improved practice regarding urine dipstick testing and appropriate urine sampling in elderly patients with possible UTI was observed during the 2019 CQUIN period. Two years later this improvement had not been sustained. When planning interventions during the 2022 CQUIN,"} +{"text": "Hyperuricemia and gout are complex diseases mediated by genetic, epigenetic, and environmental exposure interactions. The incidence and medical burden of gout, an inflammatory arthritis caused by hyperuricemia, increase every year, significantly increasing the disease burden. Genetic factors play an essential role in the development of hyperuricemia and gout. Currently, the search on disease-associated genetic variants through large-scale genome-wide scans has primarily improved our understanding of this disease. However, most genome-wide association studies (GWASs) still focus on the basic level, whereas the biological mechanisms underlying the association between genetic variants and the disease are still far from well understood. Therefore, we summarized the latest hyperuricemia- and gout-associated genetic loci identified in the Global Biobank Meta-analysis Initiative (GBMI) and elucidated the comprehensive potential molecular mechanisms underlying the effects of these gene variants in hyperuricemia and gout based on genetic perspectives, in terms of mechanisms affecting uric acid excretion and reabsorption, lipid metabolism, glucose metabolism, and nod-like receptor pyrin domain 3 (NLRP3) inflammasome and inflammatory pathways. Finally, we summarized the potential effect of genetic variants on disease prognosis and drug efficacy. In conclusion, we expect that this summary will increase our understanding of the pathogenesis of hyperuricemia and gout, provide a theoretical basis for the innovative development of new clinical treatment options, and enhance the capabilities of precision medicine for hyperuricemia and gout treatment. Gout is the leading cause of inflammatory arthritis in males. This is primarily due to multiple mechanisms resulting in the deposition of urate in the synovial fluid and other tissues to form monosodium urate crystals, which are further stimulated by inflammatory irritants, ultimately resulting in gout. The global prevalence of gout is approximately 0.1%\u201310%, and the incidence ranges from 0.3 to 6 cases per 1,000 person-years . With a The main source of uric acid is the metabolism of purines and nucleotides in food produced in the liver and excreted by the intestines and kidneys . Uric acin vivo database. This review further explores and discusses the relationship between multiple biological agents and genetic variants and how they potentially affect gout and hyperuricemia to provide a theoretical reference for further clinical treatment options.Gene variants in related functional proteins can affect uric acid metabolism and inflammation in vivo . Currentin vivo . Commonlin vivo . Severalin vivo . In addiin vivo . Althougin vivo ; howeverin vivo . This reSLC2A9), ABCG2, solute carrier family 22 member 11 (SLC22A11), solute carrier family 17 member 1 (SLC17A1), and solute carrier family 22 member 12 (SLC22A12)) was positively associated with improved renal function in European Caucasian males. The uric acid transporter protein genetic risk score was used as an instrumental variable. Mendelian randomization for renal function using the two-stage least squares method to assess the effect of urate on renal function quantitatively ) associated with blood uric acid levels and renal function in an East Asian population , and SLC22A12) , glucokinase (hexokinase 4) regulator (GCKR), SLC22A11, SLC22A12, PDZ domain containing 1 (PDZK1), and SLC17A1 were found to be significantly associated with hyperuricemia and gout risk in Asian, native Hawaiian, and Pacific Islander populations estimated using the biospecimens repository at the University of Hawai\u2019i . Previouividuals . Similar Hawai\u2019i . Most go Hawai\u2019i .HNF4A encodes a nuclear transcription factor that binds DNA and modulates the transcription of multiple genes, mainly in the form of homodimers. A missense variant in HNF4A (rs1800961) is probably the most likely leading and causal variant resulting in better transactivation of the promoter of the urate transporter protein-encoding gene ABCG2 (rs1967017) enhances HNF4A binding to the promoter of PDZK1, augmenting its expression, potentially increasing uric acid transport, and regulating uric acid homeostasis, as PDZK1 is a scaffolding protein for multiple transport proteins ((C-MAF) SNP (rs889472) might also be associated with gout susceptibility by affecting uric acid metabolism of MAF ) and the distal signal cis-eQTL (controls the expression of LINC01229). The MAFTRR lncRNA region binds to the MAF promoter and recruits the histone imprint H3K27me3 to repress MAF transcription, whereas the removal of both LINC01229 and MAFTRR promotes MAF expression can increase gout susceptibility in the Chinese population and mainly affects serum uric acid concentration and gout risk in men in the rs2941484 recessive model (k in men . In Chinve model . miR-34ave model and mighve model .ABCG2 variants (rs2231142) are variants associated with gout and an increased frequency of erythema (ABCG2 SNP (rs2231142) have a nearly 2-fold increased susceptibility to gout , the SLC2A9 SNP (rs1014290), or the SLC22A12 SNP (rs475688 and rs3825016) is linked to gout in the recessive model (ABCG2 SNP (rs2231142) significantly increased the risk of gout in Asians as well as other populations (ABCG2 SNP (rs72552713) also significantly increased the risk of gout in Asians were mainly identified as associated with the serum urate concentration or risk of hyperuricemia (ABCG2 SNP (rs2231142) enhances this autophagic impairment, diminishes the formation of neutrophil extracellular traps, and aggravates gout via the overactive release of the NLRP3 inflammasome and IL-1\u03b2. Neutrophil extracellular traps can degrade cytokines and chemokines to limit inflammation . The ABCgeneity) . ABCG2 auricemia . ABCG2 vuricemia . ABCG2 duricemia . Mitochouricemia . In addiammation . PKD2 isand gout . A transncidence .SLC16A9 SNP (rs2242206) can affect the function of its encoded monocarboxylate transporter 9 (MCT9) protein, resulting in inadequate urate excretion in the kidney are characterized as functional alleles with an approximately 6\u201310-fold greater effect on uric acid than that observed for common variants in SLC22A12 (ADH1B) SNP (rs129984) might increase the NADH/NAD ratio to promote lactate production by facilitating ethanol conversion to highly reactive acetaldehyde, thereby increasing uric acid reabsorption in synergy with the SLC22A12-encoded transporter protein URAT1 involved in eliminating endogenous and exogenous organic anions from the kidney. Tanner et al. identified multiple SNPs in SLC22A6 associated with hyperuricemia, including rs3017670, rs2276300, rs4149171, and rs4149170. Strong association studies with gout have been performed; however, there is potential evidence linking it to gout ( to gout . Granado to gout . Liu et to gout .SLC2A9 mainly encodes the GLUT9 protein. The missense variants (rs16890979) of SLC2A9 showed an association with uric acid and gout are also inconsistent based on studies on gout and hyperuricemia, and further studies are required are missense variants that serve as possible candidate causal variants for which the leucine allele leads to increased glucokinase GCK activity, resulting in increased glycolytic flux, which facilitates hepatic glucose metabolism (GCKR SNP (rs780094) is strongly associated with gout in Polynesian, European, Japanese, and Chinese populations (GCKR SNP (rs780094) was shown to be associated with the risk of hyperuricemia in men in the Uyghur population of Xinjiang in China . GCKR an= 1.311) . MLXIP etrations . MLXIPL ric acid . In addiric acid .PNPLA3 encodes an active lipase that hydrolyzes various lipids and is associated with oxidative stress (PNPLA3 SNP (rs738409) is associated with hyperuricemia in a Japanese population (PNPLA3 SNP (rs738409) enhances susceptibility to metabolism-related fatty liver disease (MAFLD) and is involved in the pathology of liver fibrosis (PNPLA3 SNP (rs738409) was associated with NAFLD in different ethnic groups in China: Han , Uyghur might correlate with blood uric acid levels by affecting the body mass index (BMI) (PNPLA3 and IGF1R variants might be linked to hyperuricemia and gout by affecting lipid metabolism and oxidative stress.e stress . A PNPLApulation . The rs7pulation . This stfibrosis . In the = 0.006) . IGF1R eex (BMI) . An abnoex (BMI) . Thus, PA1CF encodes a protein that may primarily act as an RNA binding subunit and be involved in RNA editing or processing. Rasheed et al. found that both a GCKR SNP (rs780094) and A1CF SNP (rs10821905) interact with alcohol exposure to increase the risk of gout in a European population under alcohol exposure conditions, suggesting that the involvement of GCKR and AICF in alcohol metabolism promotes the development of gout (A1CF SNP has been previously associated with hyperuricemia (A1CF SNP (rs10821905) and gout in Japanese individuals. They found that it was significantly associated with elevated serum uric acid and gout via a mechanism that might involve the regulation of dyslipidemia and uric acid metabolism are associated with blood uric acid and PDZK1 SNP (rs112129861) could interact with each other, further enriching our understanding of the genetic and biological mechanisms underlying uric acid accumulation and gout might cause the crystalline precipitation of sodium urate to trigger the inflammatory process, further exacerbating cartilage damage and promoting knee osteoarthritis, which could be associated with the inflammatory response in gouty arthritis. In addition, an interaction between an STC1 SNP (rs17786744) and GCKR SNP (rs1260326) synergistically promotes crystalline precipitation with urate-promoting gout (esponses . Studiesd levels . An STC1ing gout .HCRTR2), cytokine-dependent hematopoietic cell linker (CLNK), guanine nucleotide-binding protein a-stimulating polypeptide (GNAS)), sex hormones (breast cancer-amplified sequence 3 (BCAS3)).Various factors, such as coffee intake, tryptophan metabolism, B-cell development and activation, and sex hormones, are interlinked with genetic variants that play a role in hyperuricemia and gout. Hutton et al. found a negative association between coffee intake and gout. ABCG2, GCKR, MLXIPL, and cytochrome P450 family 1 subfamily A member 2 (CYP1A2) are variants associated with coffee consumption habits, and GCKR and ABCG2 are associated with low coffee intake and a high gout risk. Coffee consumption habits indirectly affect the association between gene variants and gout. In contrast, the direct effect of these gene variants on gout is still possible through other mechanisms, as described previously herein . EvidencHCRTR2 is a G protein-coupled receptor involved in the regulation of feeding. The encoded proteins bind to orexin A and orexin B. A HCRTR2 SNP (rs4715517), a variant associated with serum uric acid, appears to be specific to Asian populations with significantly higher allele frequencies than those in European populations. Differences in allele frequencies might contribute to interethnic differences in serum uric acid levels (CLNK SNP (rs2041215 and rs1686947) was identified as susceptibility genes for gout in the Chinese population by using dominant model and additive model , respectively (CLNK SNP (rs16869924) within the established SLC2A9 gout-associated locus was shown to increase the risk of gout in Polynesian and Chinese Tibetan individuals, genetically independent on the SLC2A9 association signal production and promotes signaling was found to be weakly associated with gout but strongly associated with blood uric acid and showed a sex-specific difference (Rs11653176 in ulations . BCAS3 culations . Studiesulations . Postmenulations . Similarspecific . An SLC1fference . It is tCLNK SNPs (rs2041215 and rs1686947) are associated with various clinicopathological parameters and might have potential as diagnostic and prognostic markers for patients with gout (ABCG2 SNP (rs2231142) respond poorly to allopurine therapy , SLC2A9 (rs16890979 and rs16891234), SLC22A11 (rs2078267), GCKR (rs1260326), matrix extracellular phosphoglycoprotein (MEPE) (rs114580333), protein phosphatase, Mg2+/Mn2+ dependent 1\u00a0K-divergent transcript (PPM1K-DT) , LOC105377323 (rs114791459), and alcohol dehydrogenase 1B (Class I), beta polypeptide (ADH1B) (rs1229984) SNPs can be used as markers of asymptomatic hyperuricemia to identify transition predictors (A case-control association study of gout in Chinese populations revealed that ith gout . Patient therapy . A GWAS edictors . HoweverGout is a form of arthritis that damages patients\u2019 physical and mental health and causes severe pain during acute attacks. Identifying individuals at risk in the early stages of the disease is essential to prevent and reduce hyperuricemia and gout and to provide pharmacological and lifestyle interventions to better treat patients with clinically diagnosed gout. The identification of genetic variants might help in disease prevention and intervention. Many GWASs have performed to uncover loci related to hyperuricemia and gout, mostly linking it to uric acid transporter proteins, such as the widely studied URAT1 and GLUT9. Some drugs have been used as targets for drug development see . We also"} +{"text": "Social distancing policies to reduce transmission of covid-19 also reduced children's exposures to endemic respiratory viruses. We aimed to examine the impact of the covid-19 pandemic on lower respiratory tract infections in under 5s presenting to primary care in England.Longitudinal trends analysis using electronic health records from a nationally representative primary care database. Our target population was children aged <5 years registered with a primary care practice from January 2015 to March 2021.Our main outcome was total weekly contacts with primary care for a lower respiratory tract infection (LRTI). We defined three pandemic phases from March 2020 - March 2021: i) first national lockdown (late March to early June 2020), ii) childcare settings reopened and second national lockdown with schools open (mid-June to mid-December 2020) and iii) third national lockdown with schools closed (late December 2020 to end of March 2021). We compared outcomes during each of the three phases with corresponding calendar weeks during pre-pandemic years 2015 to 2019.Our study population included 843 020 children <5 years who had 1 076 181 contacts with primary care for LRTIs. During the first phase (first lockdown) there were falls of 79.3% (95% CI: 73.6 to 84.5) from an average of 28 547 primary care contacts for LRTI in 2015 - 2019 to 5915 in 2020; there was a 78.9% (95% CI: 73.7 to 83.9) fall in phase two (childcare settings reopened and second lockdown) from 107 873 to 22 792 contacts; and a 77.7% (95% CI: 73.5 to 81.4) fall in phase three (third lockdown) from 57 200 to 12 764 contacts.Children under 5 in England had fewer contacts with primary care for LRTIs during the covid-19 pandemic. This change likely reflects lower prevalence of respiratory illness due to fewer social contacts. This may impact on future health service use as these children have had less exposure, and therefore may have less immunity, to respiratory diseases.\u2022\u2002Children under 5 had fewer contacts with primary care for lower respiratory tract infections during the covid-19 pandemic in England likely due to the restrictions in place to reduce social contacts.\u2022\u2002The falls in lower respiratory tract infections during the covid-19 pandemic in under 5s may mean they have less immunity to respiratory viruses which may impact upon their future health service use."} +{"text": "Primary cilia play counterregulatory roles in cystogenesis\u2014they inhibit cyst formation in the normal renal tubule but promote cyst growth when the function of polycystins is impaired. Key upstream cilia-specific signals and components involved in driving cystogenesis have remained elusive. Recent studies of the tubby family protein, Tubby-like protein 3 (TULP3), have provided new insights into the cilia-localized mechanisms that determine cyst growth. TULP3 is a key adapter of the intraflagellar transport complex A (IFT-A) in the trafficking of multiple proteins specifically into the ciliary membrane. Loss of TULP3 results in the selective exclusion of its cargoes from cilia without affecting their extraciliary pools and without disrupting cilia or IFT-A complex integrity. Epistasis analyses have indicated that TULP3 inhibits cystogenesis independently of the polycystins during kidney development but promotes cystogenesis in adults when polycystins are lacking. In this review, we discuss the current model of the cilia-dependent cyst activation (CDCA) mechanism in autosomal dominant polycystic kidney disease (ADPKD) and consider the possible roles of ciliary and extraciliary polycystins in regulating CDCA. We then describe the limitations of this model in not fully accounting for how cilia single knockouts cause significant cystic changes either in the presence or absence of polycystins. Based on available data from TULP3/IFT-A-mediated differential regulation of cystogenesis in kidneys with deletion of polycystins either during development or in adulthood, we hypothesize the existence of cilia-localized components of CDCA (cCDCA) and cilia-localized cyst inhibition (CLCI) signals. We develop the criteria for cCDCA/CLCI signals and discuss potential TULP3 cargoes as possible cilia-localized components that determine cystogenesis in kidneys during development and in adult mice. PKD1 or PKD2, which encodes polycystin-1 (PC1) or polycystin-2 (PC2), respectively (polycystins or PCs collectively) is characterized by the formation of numerous fluid-filled cysts from allctively) . PC1 is ctively) with a Gctively) . PC1 undctively) . PC2 belctively) and can ctively) . PC1 andctively) . Recent ctively) . Moreovectively) . Polycysctively) by unknoctively) , mTOR (Sctively) , cAMP (Tctively) , WNT (Kictively) , Ca2+ (Kely) (2+ and G-prely) (2+ are dysrely) mice, the Somlo laboratory has proposed an unidentified cilia-dependent cyst activation (CDCA) signal(s) . The CDCignal(s) . The funignal(s) . Loss oftivation . This letivation . Ciliary mutants .Pkd1 influenced the progression of polycystic kidney disease in mouse Pkd1 models of developmental or adult-onset of ADPKD. These results suggested that the Hedgehog signaling pathway does not contribute to the CDCA or other ciliary signals that drive renal cystogenesis.The molecular nature of CDCA components and effectors are unknown. The CDCA mechanism of cyst growth is active in all tubular segments following both early and late ADPKD gene inactivation . The CDCPkd1 . In factPkd2 null-like phenotype characterized by embryonic kidney cysts expressed in the cell body remains completely Endoglycosidase H (Endo H) sensitive implying that this mutant is retained in the ER and defective in the trafficking , albeit less effectively than the wild-type protein. It remains to be seen whether the anti-cystogenic role of PC1V in the cell body is mediated through its function in the mitochondria . Without cilia, the cilia-dependent signal should be abolished and the cyst inhibiting role of polycystins should prevail. Second, cyst growth continues to persist in Pkd1-cilia double mutants ; however, loss of cilia should prevent CDCA from initiating cystogenesis if the cyst activation signal solely originates in the cilium. Moreover, neither concomitant reduction in dose of PC1 nor transgenic overexpression of PC1 were found to have an impact on the cystic burden in cilia mutants . An alternative parsimonious interpretation would be that persistent cyst growth in Pkd1-cilia double mutants arises from additional ciliary roles independent of polycystins. Cilia-defective mutants are likely to cause fibrocystic kidney disease phenotype by mechanisms signal(s).The above observations have additional implications. First, and according to the model Figure , the CDCop cysts , \u201c?\u201d. Anchanisms that arechanisms . The cilOverall, these considerations suggest a more complex cilia-regulated mechanism in cyst growth involving a combination of positive and negative regulatory signals in ADPKD. These counterregulatory signals generated within the cilium are in a finely tuned balance to facilitate functional tubule adaptation to physiological inputs in normal kidneys. Dysregulation of these ciliary signals may underlie cystic kidney diseases.Several difficulties have limited our ability to dissect the crucial ciliary signals. First, ciliary signals cannot be identified in experimental models that lack cilia. Ablation of cilia, an important cellular compartment, disrupts a great number of cellular pathways, including cell cycle regulation and cell polarity . The cruPkd1 single and Pkd1-cilia double mutant kidneys has identified non-ciliary cyclin-dependent kinase 1 as a driver of cyst cell proliferation from Pkd1 inactivation but did not find changes in ciliary drivers in cystogenesis ciliary delivery by IFT-A core-binding to TULP3 N-terminus, and 3) release into PIP2-deficient ciliary membrane is a key adapter of the intraflagellar transport complex A (IFT-A) in the trafficking of multiple proteins specifically into the ciliary membrane. The IFT-A holo-complex is generally considered to be regulating the retrograde trafficking of cargoes including IFT-B complex in the cilia . TULP3 imembrane of ary loss and can be defined experimentally by the following criteria:(i) It is trafficked to cilia.(ii) It could be a ciliary cargo of TULP3.Pkd1/2 cko.(iii) Lack causes cystic changes but milder than Pkd1/2 cko enhances cystic kidney phenotype during development.(iv) Concomitant cko with Based on these results, we propose that a subset of TULP3/IFT-A ciliary cargoes generate Tulp3 and Pkd1 in adult mouse kidneys have provided compelling evidence for a critical role of Tulp3 in the trafficking of the ciliary component(s) of the CDCA signal in Pkd1 cko mice. The Liem laboratory showed that in adult mice, concomitant loss of Tulp3 completely suppressed cystogenesis in Pkd1 cko in adult mice at 18 weeks of the CDCA signal and delineate those as a subset of Tulp3/IFT-A ciliary cargoes in adult kidneys. To highlight their cilia-specificity and distinctiveness in action within the cilia, we term the cilia-localized component ciliary CDCA or cCDCA.These results suggest that TULP3 traffics cilia localized cyst promoting signal(s) into cilia, which are suppressed by polycystins in normal tubules but are derepressed in adult-onset Pkd1 cko late-onset model offers a cyst suppressor system to test the cCDCA candidates from among the TULP3/IFT-A ciliary cargoes. The TULP3-dependent cCDCA signal(s) may be identified experimentally by the following criteria:(i) It is trafficked to cilia.(ii) It is a ciliary cargo of TULP3.(iii) Lack causes no cystic changes in early adulthood.Pkd1/2 cko at early adulthood.(iv) Concomitant cko rescues adult-onset PKD in The Tulp3 for the IFT-A peripheral subunit Ift139/Thm1. Concomitant loss of this IFT-A subunit suppresses cystogenesis from PC1 or PC2 loss in adult kidneys while retaining the cilia and negatively (CLCI) impact cyst growth . In the adult kidney, TULP3/IFT-A might predominantly traffic the cCDCA component(s) unique to the adult state, but few CLCI components (until the later adult stages) . The CLCPkd1. Conditional inactivation of Tulp3 causes a stop to the ongoing ciliary trafficking of the cCDCA components. However, the already elevated cCDCA may have delayed turnover from the cilia to halt rapid cyst growth following early Pkd1 inactivation. Residual activity of cCDCA is likely sufficient to drive considerable cyst growth that is rapidly ongoing, thus preventing a rescue of cyst growth in the Tulp3-Pkd1 double cko during development. This scenario is consistent with the previous finding that severity of cyst growth is highly sensitive to the length of time between the initial loss of the polycystins and the subsequent involution of cilia . In comparison, cysts grow at a much slower rate following Pkd1 cko in adult age. The elevated level of cCDCA may drop below a threshold that is required to promote cyst growth within a time interval that is not sufficient to drive cyst growth to a significant extent.Alternatively, there may be no fundamental differences in the TULP3/IFT-A regulated trafficking of cilia-signaling components between the two stages. The cCDCA may instead be trafficked by TULP3/IFT-A during development as in adult kidneys, but this regulation may be obscured by the rapid cyst growth following early inactivation of of cilia . TherefoSuch counterregulatory ciliary roles are the foundational basis of the Hedgehog (Hh) pathway . We and Pkd1 mutants of varying severity during kidney development or in adult kidneys would inform whether TULP3 cargoes function as likely CLCI or cCDCA signals. A decrease in the severity of Pkd1 mutant cystic phenotype upon concomitant loss of a TULP3 cargo would argue for this cargo to function as a cCDCA signal. An increase in severity of Pkd1 mutant cystic phenotype upon concomitant loss of a TULP3 cargo would argue for this cargo to function as a CLCI signal.The genetic epistasis approaches with TULP3 cargoes and Tulp3 deletion alone causes milder cystogenesis than Pkd1 loss. Concomitant Tulp3 cko in Pkd1 cko animals did not inhibit cystogenesis but caused earlier lethality than Pkd1 cko alone, suggesting that Tulp3 inactivation accelerated loss of renal function in the Pkd1 cko. Therefore, it is highly likely that TULP3 traffics cilia-localized cyst inhibition (CLCI) signal(s) during kidney development. Alternatively, Tulp3 could additionally regulate a cCDCA signal during development, but fast cyst progression in absence of Pkd1 could make such regulation difficult to detect. Genetic epistasis between TULP3 cargo and developmental models of Pkd1 mutants of varying severities could unmask either signal.Pkd1-Tulp3 double cko mice is fully suppressed at this stage. Intact kidney epithelial cilia in Tulp3 mutants argue for cilia-generated signaling rather than gross ciliary morphology defects in such suppression. Thus, it is highly likely that TULP3 traffics the ciliary component of CDCA (cCDCA) signal(s) predominantly in adult kidneys. Mild cystogenesis from Tulp3 deletion in adult-onset models occurs much later at 42 weeks, suggesting low CLCI activity of TULP3 cargoes only at older ages. Genetic epistasis between Tulp3 cargo mutants and Pkd1 mutants could therefore unmask cCDCA signal(s).Lack of TULP3 in adult-onset models does not cause cystogenesis at 18 weeks. Cystogenesis in adult-onset Tulp3 cko that shows a selective loss of the corresponding TULP3 cargo from cilia alone without affecting the extraciliary pools. In certain cases, TULP3 cargoes that are selectively deficient in trafficking to cilia without affecting the functionality can be generated by mutating ciliary localizing signals targeted by TULP3 independent of polycystins during development and cCDCA signal(s) repressed by polycystins in adult kidneys.in vivo (Arl13b causes mild fibrocystic kidney disease in mice models (arl13b (scorpion) allele has nephric duct dilatation phenotypes, and analysis of phenotypic rescue using arl13b variants in this model suggests that ciliary localization is essential for in vivo function of ARL13B , and myristoylated proteins by releasing them from their binding partners PDE6\u03b4 shows cystogenesis in the embryonic kidney, but cilia are maintained (Tulp3 K407I mutant (Pkd2lrm4 (E442G) mutant in the first extracellular loop is unable to localize to cilia despite an intact RVxP shows reduced AC3 in the neural tube cilia (Both AC5 and AC6 AC5 (AC6 deletionAC5 (AC6 . AC5/6 aAC5 (AC6 . The autbe cilia . AC3 is be cilia . Whetherbe cilia could beTulp3 cko kidney epithelial cilia (2 (2, it could regulate ciliary components. The CLCI signal could be activating a transcription factor in cilia that is regulatable by the ciliary microenvironment. Such an example is seen in Hh pathway where GLI2 and GLI3 transcription factors are modified in cilia upon Hh addition (Kif3a loss (What could be the molecular output propagated in cilia by CLCI and cCDCA signals? Based on our results showing early depletion of ARL13B and INPP5E from al cilia , they arcilia (2 . If INPPaddition . Some ofaddition , at leasaddition and GLISaddition , are ciladdition or in thf3a loss . The cCDf3a loss . Nonethe"} +{"text": "Cladocopium C1acro strain. Genotyping of the Symbiodiniaceae communities before bleaching and after reinoculation showed that fragments of all six coral species acquired the Cladocopium C1acro strain used for inoculation. Our results provide strong evidence for the uptake of Symbiodiniaceae from the environment by adult corals. We also demonstrate the feasibility of chemical bleaching followed by reinoculation to manipulate the Symbiodiniaceae communities of adult corals, providing an innovative approach to establish new symbioses between adult corals and heat-evolved microalgal symbionts, which could prove highly relevant to coral reef restoration efforts.Early life stages of most coral species acquire microalgal endosymbionts (Symbiodiniaceae) from the environment, but whether exogenous symbiont uptake is possible in the adult life stage is unclear. Deep sequencing of the Symbiodiniaceae ITS2 genetic marker has revealed novel symbionts in adult corals following bleaching; however these strains may have already been present at densities below detection limits. To test whether acquisition of symbionts from the environment occurs, we subjected adult fragments of corals (six species in four families) to a chemical bleaching treatment (menthol and DCMU). The treatment reduced the native microalgal symbiont abundance to below 2% of their starting densities. The bleached corals were then inoculated with a cultured Scleractinian corals prosper in oligotrophic waters by forming mutualistic relationships with microalgae (Symbiodiniaceae) that translocate photosynthate to their host . BreakdoDurusdinium have been found to increase the bleaching threshold of the coral holobiont by 1\u20132\u2009\u00b0C [Bleaching tolerance of corals to elevated temperatures varies within and among species. This is partly determined by the physiological performances of their microalgal symbionts under thermal stress , 5. For y 1\u20132\u2009\u00b0C .Porites divarcata following bleaching [The Symbiodiniaceae comprise at least 15 genera and genus-level lineages which include many species . SeveralDiploastrea heliopora (Diploastraeidae), Dipsastraea pallida (Merulinidae), Echinopora lamellosa (Merulinidae), Platygyra daedalea (Merulinidae), Porites lobata (Poritidae) and Stylophora pistillata (Pocilloporidae)), which were then successfully reinfected with a cultured Symbiodiniaceae strain. Colonies were fragmented and chemically bleached (n\u2009=\u200916 per species) through exposure to menthol and 3--1,1-dimethylurea : (1) a negative control treatment (Ctl-) where corals were not reinoculated with any Symbiodiniaceae; (2) a positive control treatment (Ctl+) where corals were reinoculated with freshly isolated homologous Symbiodiniaceae ; (3) a reinoculation treatment (Ri) where corals were reinoculated with a cultured Cladocopium C1acro strain (SCF055-01.10) at 104 cells/ml; (4) a reinoculation with sand treatment (RiS) where corals were reinoculated as in treatment Ri in the presence of sterilised sand region of the Symbiodiniaceae nrDNA was undertaken to characterise the symbiont communities before bleaching and nine weeks after the first reinoculation with ro Table\u00a0. The nat region oCladocopium C1acro strain by P. lobata and S. pistillata contributes to the growing body of evidence that vertically transmitting corals also possess the ability to acquire symbionts horizontally [P. daedalea only, which led to a mixed community with a Durusdinium phylotype that was initially detected in low abundance (<1%).The environmental acquisition of the zontally . The preStylophora pistillata and Isopora palifera corals have been found to lose 99% of their algal symbiont densities following menthol exposure [Exaiptasia diaphana [Chemical bleaching is increasingly used in studies investigating cnidarian-symbiont interactions to obtain aposymbiotic hosts for reinfection with cultured Symbiodiniaceae , 18. Aduexposure . ComparaP. lobata) and robust clades, can acquire heterologous Cladocopium C1acro symbionts from the environment. The chemical bleaching and reinoculation methodology may prove highly relevant for the study of coral-Symbiodiniaceae interactions and for the development of adult coral stock with enhanced thermal tolerance for reef restoration by inoculating them with heat-evolved Symbiodiniaceae [Our findings provide compelling evidence that adult fragments of six coral species, spanning four families in the complex (iniaceae .Supplementary InformationDataset 1"} +{"text": "The scRNA-seq data of bone marrow mononuclear cells from 17 healthy donors (HDs) were downloaded from the Gene Expression Omnibus database (GSE120221) [Myelodysplastic syndromes (MDS) are heterogeneous clonal diseases characterized by cytopenia caused by ineffective hematopoiesis and high risk of transformation into acute myeloid leukemia (AML). At present, the pathogenesis of MDS has not been elucidated. MDS is a group of stem cell diseases. The abnormal proliferation and blockade in differentiation of hematopoietic stem cells (HSCs) result in cytopenia and leukemic transformation. HSCs architectures in MDS can also predict therapeutic reaction . Therefo\u2212 hematopoietic cells were analyzed. After dimension reduction, clustering, and visualization, we identified HSC/multipotent progenitor (MPP) populations and conducted an in-depth analysis. We investigated the properties of MDS stem cells by analyzing the differentially expressed genes (DEGs) between patients and HDs. We found that genes associated with neutrophil granule, such as MPO, AZU1, DEFA3, had elevated expression in MDS patients compared with HDs , HR-MDS (n\u2009=\u200940) and sAML (n\u2009=\u200925) were significantly lower than those of control (n\u2009=\u200940) (P\u2009<\u20090.001) is related to the loss of heterozygosity of q is relaBelow is the link to the electronic supplementary material.Supplementary Material 1Supplementary Material 2Supplementary Material 3"} +{"text": "Antibiotic use contributes to antibiotic resistance affecting individual patients and communities. Important antimicrobial stewardship program (ASP) strategies, such as prospective audit and feedback, may be limited to peak weekday hours in many institutions. Currently, a lack of data exists to support expansion of ASP beyond peak hours in community hospitals. The goal of this study was to describe the impact of expanding inpatient ASP weekend coverage with a newly established PGY2 infectious diseases pharmacy resident program.This retrospective cohort study was approved by the Institutional Review Board and conducted using the pharmacist documentation function within the electronic health record of weekend interventions taking place between July 1, 2021 and December 31, 2022. The primary objective was to describe the impact of expanding weekend AMS coverage with a PGY2 ID pharmacy resident through quantification of inpatient antimicrobial stewardship interventions. Secondary endpoints included comparing the PGY2 resident\u2019s weekend stewardship activities based on intervention quantity, type, and impact, to weekends without PGY2 ID resident coverage. Comparator groups included: 1) A single experienced clinical pharmacist, 2) Two new PGY1 pharmacy residents (first 6 months of residency), and 3) Two experienced PGY1 pharmacy residents .8 weekends of interventions were collected for each group. Significantly more interventions, including low-, medium-, and high-impact interventions were made by an ID-PGY2 pharmacy resident compared to a clinical pharmacist and both PGY1 groups. The median number of interventions made by each group are outlined in Table\u00a01. Additionally, significantly more interventions were made per protocol as well as interventions that required communication with providers.Expansion of ASP services to include weekend clinical coverage with an ID PGY2 pharmacy resident significantly increased weekend ASP interventions in a community teaching hospital. Specialty pharmacy residency training programs may offer a high-value opportunity to expand ASP coverage.All Authors: No reported disclosures."} +{"text": "Background: Does the presence of single-nucleotide polymorphisms (SNPs) in the leukemia inhibitory factor (LIF) gene affect ovarian response in infertile young women? Methods: This was a case\u2013control study recruiting 1744 infertile women between January 2014 to December 2015. The 1084 eligible patients were stratified into four groups using the POSEIDON criteria. The gonadotropin-releasing hormone receptor (GnRHR), follicle-stimulating hormone receptor (FSHR), anti-M\u00fcllerian hormone (AMH), and LIF SNP genotypes were compared among the groups. The distributions of LIF and FSHR among younger and older patients were compared. Clinical outcomes were also compared. Results: The four groups of poor responders had different distributions of SNP in LIF. The prevalence of LIF genotypes among young poor ovarian responders differed from those of normal responders. Genetic model analyses in infertile young women revealed that the TG or GG genotype in the LIF resulted in fewer oocytes retrieved and fewer mature oocytes relative to the TT genotypes. In older women, the FSHR SNP genotype contributed to fewer numbers of mature oocytes. Conclusions: LIF and FSHR SNP genotypes were associated with a statistically significant reduction in ovarian response to controlled ovarian hyperstimulation in younger and older women with an adequate ovarian reserve, respectively. Leukemia inhibitory factor (LIF) is a cytokine belonging to the interleukin-6 superfamily. It was first identified for its ability to induce macrophage differentiation of murine myeloid leukemia cells and inhibit their proliferation ,2. LIF mOocyte quantity and quality are crucial for fecundability . Oocyte Maternal age-related aneuploidy and euploidy affecting oocyte quality are another set of factors that influence ART prognosis ,13. HoweFSHB; ) and 3 (64.1% [42.7% plus 21.4%]) than in the older women in group 2 (57.4% [43.8% plus 13.6%]) and 4 . No such result was observed for other SNP genotypes, such as GnRHR (rs3756159), FSHR (rs6166), and AMH (rs10407022) (The distributions of SNPs for the groups are presented in 0407022) .FSHR (rs6166) A allele frequencies were higher in women with adequate ovarian reserves and with a suboptimal or poor response after conventional COH (group 1 [69.7%] and 2 [69.4%]) than women with a poor ovarian reserve . The distribution of G allele frequencies of LIF (rs929271) were significantly higher among young women in group 1 (42.3%) and 3 (42.7%) than among older women in group 2 (35.3%) and 4 . No such result was observed for allele frequencies of other SNPs, such as GnRHR (rs3756159) and AMH (rs10407022) genes. These results suggest an association of an SNP in the LIF (rs929271) with low-prognosis groups, especially in patients younger than 35 years. Furthermore, an association of the A allele of FSHR (rs6166) with a suboptimal or poor ovarian response during conventional COH was observed, especially in patients with an adequate ovarian reserve.LIF (rs929271) TG/GG genotypes and G allele were enriched in young patients and that the A allele frequency of FSHR (rs6166) was higher in POSEIDON groups 1 and 2. Thus, we compared the LIF (rs929271) and FSHR (rs6166) genotypes and allele frequencies of POSEIDON group 1 (n = 208) and group 2 (n = 361) with those of age-matched normal responders .Patients in POSEIDON group 1 or 2 exhibited an unexpectedly poor or suboptimal ovarian response after standard ovarian stimulation, despite having adequate ovarian reserve parameters . The resLIF (rs929271) and FSHR (rs6166) across the various ages and groups is presented in LIF (rs929271) results indicated significant differences between the genotypes of LIF (rs929271) in poor-responder groups and in young (age < 35 years) and normal responders; the TG/GG and G alleles was more common in group 1 than in normal responders . This distribution did not significantly differ between group 2 (age \u2265 35 years) and normal responders (The distribution of sponders .FSHR (rs6166), the A allele was more common in older women than in normal responders . However, the FSHR (rs6166) genotypes were not more common in women older than 35 years (FSHR (rs6166) allele frequency and genotypes were similar between patients with poor response and individuals in the control group.With regard to 35 years . In womeLIF (rs929271) with poor response, especially in patients younger than 35 years. The allele frequencies of FSHR (rs6166) were associated with poor response, especially in women older than 35 years.Overall, these results demonstrated an association of SNPs in the LIF (rs929271) TG/GG genotypes were more common in younger patients with poor response than in normal responders. The influence of LIF (rs929271) genotypes on clinical characteristics and clinical outcomes was investigated in patients younger than 35 years undergoing ART treatment.Our results revealed that the A total of 599 women (age < 35 years) were included in genetic model analysis, and the results are displayed in LIF (rs929271) did not differ with respect to age; BMI; AMH; baseline FSH, LH, and E2; duration of infertility; E2 on human chorionic gonadotrophin (HCG) administration day; P4 on HCG administration day; number of D3 embryos; or D3 good embryo rate (LIF (rs929271) TG/GG genotype retrieved significantly fewer oocytes than those with the TT genotype gene was also associated with a significantly lower number of mature oocytes for the genotype TG/GG than that for the TT genotype may contribute to decreases in the number of oocytes retrieved and the number of mature oocytes in young women with infertility younger than 35 years undergoing ART treatment.The patients\u2019 clinical characteristics between the TT and TG/GG genotypes of the ryo rate . HoweverFSHR (rs6166) were associated with the POSEIDON group 2 patients (age \u2265 35 years), the effects of the FSHR (rs6166) genotypes on clinical characteristics and clinical outcomes were also examined in older patients (age \u2265 35 years) undergoing ART treatment.Because the allele frequencies of A total of 630 women older than 35 years were included in genetic model analysis ; 564 (89FSHR (rs6166) did not significantly differ with respect to age; BMI; AMH; baseline FSH, LH, and E2; duration of infertility; E2 on HCG administration day; P4 on HCG administration day; number of oocytes retrieved; number of D3 embryos; or D3 good embryo rate (FSHR (rs6166) AA/AG genotype had significantly fewer mature oocytes than women with a GG genotype may lead to lower numbers of mature oocytes in older women with infertility (age \u2265 35 years) undergoing ART treatment.The patients\u2019 clinical characteristics between the AA/AG and GG genotypes of the ryo rate . HoweverLIF (rs929271) TG/GG genotypes and G allele. A higher frequency of FSHR (rs6166) A allele was observed in the women with infertility with adequate ovarian reserve (POSEIDON group 1 and 2). Second, we compared the distribution of LIF (rs929271) and FSHR (rs6166) in patients with infertility with an adequate ovarian reserve (POSEIDON group 1 and 2) with normal responders. The women with infertility under the age of 35 (POSEIDON group 1) were associated with a higher frequency of TG/GG genotypes and G allele of LIF (rs929271). The older women with infertility were associated with a higher A allele frequency of FSHR (rs6166). Finally, we demonstrated that LIF (rs929271) may lead to fewer oocytes retrieved and a lower number of mature oocytes in young women with infertility under the age of 35 years and the FSHR (rs6166) may contribute to fewer number of mature oocytes in older women with infertility (age \u2265 35) undergoing ART treatment. According to our results, LIF (rs929271) and FSHR (rs6166) were associated with a statistically significant reduction in ovarian response to controlled ovarian stimulation (COH) in younger and older women, respectively, with an adequate ovarian reserve. These results indicated that both LIF (rs929271) and FSHR (rs6166) might modulate ovarian response during COH.We first evaluated the distribution of four SNP polymorphisms in POSEIDON groups. We found that women with infertility under the age of 35 (POSEIDON group 1 and 3) were associated with a higher frequency of LIF (rs929271) are significantly enriched in patients with infertility under the age of 35 years, but not in older patients with unexplained infertility [LIF (rs929271) in younger patients with infertility (group 1 and 3) but not in older patients (group 2 and 4) among the patients with poor response.One study demonstrated that GT/GG genotypes and the G allele of ertility . Our stuFSHR (rs6166) were similar when we compared POSEIDON group 3 with group 4 (low ovarian reserve) [FSHR (rs6166) among POSEIDON groups, our current study indicated a higher frequency of A allele of FSHR (rs6166) in the women with infertility with adequate ovarian reserve A allele than women with infertility with low ovarian reserve. However, for women with infertility with low ovarian reserve, the frequency of A allele of FSHR (rs6166) was not distributed differently.Our previous study indicated that the frequencies of the SNP in reserve) . HoweverLIF (rs929271) was distributed differently in POSEIDON group 1 and the control group. A higher frequency of A allele of FSHR (rs6166) was noted in POSEIDON group 2 than the control group (The number of oocytes retrieved following COH for IVF/ICSI is closely related to cumulative live birth rates (LBR) after utilization of all fresh and frozen embryos . High reol group . Our resLIF (rs929271) was associated with a significantly lower number of oocytes (14 vs. 16) and mature oocytes (11 vs. 13). These results were reflective of the higher percentages of TG/GG genotypes (69.2%) in POSEIDON group 1 than in normal responders (58.3%) with AMH \u2265 1.2 ng/mL, a genetic model analysis revealed that compared with a TT genotype, a TG or GG genotype in (58.3%) . These eFSHR (rs6166) was associated with a significantly lower number of mature oocytes (8 vs. 10). The higher frequency of A allele of POSEIDON group 2 than in normal responders with AMH \u2265 1.2 ng/mL, a genetic model analysis revealed that compared with a GG genotype, an AA/AG genotype in sponders may haveLIF polymorphisms among women younger than 35 years with unexplained infertility [LIF (rs929271) may exhibit unexpectedly poor or suboptimal responses during COH, and LIF may affect folliculogenesis both in gonadotropin-independent growth and gonadotropin-dependent growth.Numerous studies have investigated the ertility , as predertility and in tertility . In animertility , enhanceertility , and modertility . LIF mayFSHR polymorphisms have been investigated in relation to ovarian response. The earliest report on this topic indicated that more FSH ampoules are required to reach successful stimulation when the G/G genotype of FSH (rs6166) is present at a significantly higher basal level [FSHR (rs6166) A/A and rs6165 G/G genotypes and rs1394205 A/A genotype tend to exhibit reduced ovarian response during COH and require higher FSH dosages [FSHR and FSHB genes on ovarian response and reported that the presence of FSHR SNPs affected ovarian responses with a fixed dose of 150 IU rFSH [FSHR SNPs affect folliculogenesis in gonadotropin-dependent growth. Therefore, fewer oocytes are retrieved by older patients (\u226535 years) with AA or AG genotypes of FSHR (rs6166) with AMH \u2265 1.2 ng/mL receiving ART treatment.al level . Numerou dosages ,32,33,34 IU rFSH . These fLIF (rs929271) and FSHR (rs6166) analysis should be considered for young and older women with infertility who are expected to be normal responders but who exhibit an unexpectedly poor or suboptimal COH response.With regard to the clinical implications of our findings, LIF (rs929271). The older women with infertility were associated with a higher A allele frequency of FSHR (rs6166). Infertile patients in POSEIDON group 1 and 2 have some common features, such as an adequate ovarian reserve test (AFC \u2265 5\u20137 follicles or AMH \u2265 1.2 ng/mL), but they revealed unexpected impaired or poor ovarian response to exogenous gonadotropins undergoing COH. Thereafter, we could consider genotyping LIF (rs929271) and FSHR (rs6166) among young and older women with infertility who are expected to be normal responders before they enter their first ART to avoid an unexpected or a hypo-response. This study has two major strengths: (a) the use of only one type of long GnRH agonist stimulation protocol and (b) the strict inclusion criteria for patients with poor response based on the POSEIDON criteria.It will be a challenge for reproductive specialists to predict impaired or poor ovarian response to exogenous gonadotropins when infertile women with an adequate ovarian reserve test (AFC \u2265 5\u20137 follicles or AMH \u2265 1.2 ng/mL) undergo their first ART. Our data showed that the women with infertility under the age of 35 (POSEIDON group 1) were associated with a higher frequency of TG/GG genotypes and G allele of LIF (rs929271) or FSHR (rs6166) influences the rate of clinical pregnancy.This study has several limitations. First, we included only Han Chinese people. Second, we did not include data regarding pregnancy outcomes after embryo transfer. Therefore, we cannot conclude that LIF (rs929271) and FSHR (rs6166) were associated with a statistically significant reduction in ovarian response to COH in younger and older women, respectively, with an adequate ovarian reserve, indicating that both LIF (rs929271) and FSHR (rs6166) might modulate ovarian response during COH. LIF (rs929271) and FSHR (rs6166) should be considered as potential biomarkers for poor or suboptimal COH responses among young and older women with infertility who are expected to be normal responders."} +{"text": "National Comprehensive Cancer Network (NCCN) guidelines recommend HAIP chemotherapy for advanced iCCA confined to the liver only in experienced centers or clinical trials.4 In this meta-analysis, we investigated OS after HAIP chemotherapy with floxuridine for patients with unresectable iCCA.Patients with advanced cholangiocarcinoma had median overall survival (OS) of only 1 year with palliative gemcitabine\u2013cisplatin in the Advanced Biliary Cancer (ABC) trials. No survivors were observed beyond 3 years.5 These results compare favorably with the results of systemic chemotherapy reported in the ABC trials.2 However, no randomized controlled trial (RCT) has been performed.Nine studies, including three phase II trials, had a total sample size of 478 patients without extrahepatic disease other than locoregional nodal disease, of whom 154 unique patients were included in the meta-analysis. The weighted median OS was 29.0 months, and the pooled 3-year OS was 39.5%.More research is necessary to optimally determine the efficacy of HAIP chemotherapy for unresectable iCCA, and additional study of quality-of-life measures is recommended. An international RCT (NCT04891289) is currently investigating the additional value of HAIP chemotherapy for patients with unresectable iCCA treated with palliative gemcitabine\u2013oxaliplatin (GemOx)."} +{"text": "Memory and learning deficits are central among cognitive deficits in schizophrenia. However, to a varying proportion ca. 20-25% of patients could not be considered deficit.Description of sociodemographic, lifestyle and clinical factors related to good performance in PAL-test in schizophrenia patients.th\u00a0percentile scores (10 error score or less) for outcome measure total errors adjusted (TEA) of NFBC 1966 was used as a cut-off for good performance in PAL test.Participants (N=4500) were members of the Finnish SUPER study on the genetic mechanisms of psychotic disorders (SUPER). The database of the Northern Finland Birth Cohort 1966 (NFBC 1966) was utilized as a reference data. Visual memory and new learning were assessed using Cambridge Neuropsychological Test Automated Battery (CANTAB) Paired Associates Learning (PAL) test. The 50The sociodemographic and lifestyle factors related good performance for both sexes were: younger age (p<.001), higher basic education (p <.001), independent form of dwelling (p<.001), hazardous drinking (p <.001), cannabis use (p <.001) and being married . The clinical factors related to good performance for both sexes were not using antipsychotic medication regularly (p <.001), not using all psychotropic medication , less hospitalization times due to psychosis (p <.001), younger age at first hospitalization due to psychosis (p <.001), lower number of hospitalization days (p <.001) and lower percentage of time in hospital after first psychosis episode (p <.001).Several factors related to good performance in the PAL\u2013test in the crude analysis without any confounders."} +{"text": "PPD is associated with a range of adverse outcomes for both the mother and infant2. Therefore, identifying modifiable risk factors for perinatal depression is an important public health issue3.Postpartum depression (PPD) is the commonest postpartum psychiatric condition, with prevalence rates around 20%To explore the role of dysfunctional attitudes towards motherhood in postpartum depressive symptoms and disorder.4, the Postpartum Depression Screening Scale5\u00a0and the Diagnostic Interview for Psychological Distress-Postpartum6. Correlation analysis was performed followed by linear/logistic regression analysis when the coefficients proved significant (p<.05), using SPSS.247 women were evaluated in the third (12.08\u00b14.25 weeks) and sixth months (31.52\u00b1 7.16 weeks) postpartum with the Attitudes Towards Motherhood ScaleDysfunctional beliefs towards motherhood concerning judgement by others and maternal responsibility positively correlated with depressive symptoms at the third and the sixth months postpartum. Those dysfunctional beliefs were predictors of depressive symptoms at the third and sixth months explaining 29.4% and 30.2% of its variance, respectively. Having dysfunctional beliefs at the third month significantly increase the likelihood of being diagnosed with Major Depression (DSM5) both in the third and sixth months .Cognitive distortions should be included in the assessment of risk factors for PPD. Early identification of women presenting motherhood-specific cognitive biases may be crucial for implementing preventive interventions favoring a more positive and healthier motherhood experience.No significant relationships."} +{"text": "Rhododendron.Many diverse plant clades possess bilaterally symmetrical flowers and specialised pollination syndromes, suggesting that these traits may promote diversification. We examined the evolution of diverse floral morphologies in a species\u2010rich tropical radiation of Rhododendron sect. Schistanthe to reconstruct phylogenetic relationships and examine hybridisation. We then captured and quantified floral variation using geometric morphometric analyses, which we interpreted in a phylogenetic context.We used restriction\u2010site associated DNA sequencing on 114 taxa from Rhododendron species that began diversifying into New Guinea c. 6\u2009million years ago expanded into novel floral morphological space.We uncovered phylogenetic conflict and uncertainty caused by introgression within and between clades. Morphometric analyses revealed flower symmetry to be a morphological continuum without clear transitions between radial and bilateral symmetry. Tropical Rhododendron is characterised by recent speciation, recurrent hybridisation and the origin of floral novelty. Floral variation evolved via changes to multiple components of the corolla that are only recognised in geometric morphometrics with both front and side views of flowers.Our results showed that the evolution of tropical Flower symmetry, in particular bilateral symmetry in which a flower exhibits one plane of symmetry along the dorsoventral axis, is believed to facilitate pollinator attraction, visitation, efficiency, precision and specialisation is a large group of over 1000 species, with notable species richness in the tropics. This diversity arose via southward dispersal from the group\u2019s origin in Northeast Asia, which is likely to have been facilitated by mountain building in the Malay Archipelago or curved abaxially (upwards) Fig.\u2009 (Stevensds) Fig.\u2009. The onlds) Fig.\u2009 (Sleumerds) Fig.\u2009. Howeverds) Fig.\u2009 descriptSchistanthe. Specifically, we aimed to (1) resolve the phylogeny of Rhododendron sect. Schistanthe using genome\u2010wide data, (2) investigate whether hybridisation underlies phylogenetic complexity, and (3) characterise flower symmetry variation and evolution in this species radiation. We used restriction\u2010site associated DNA sequencing (RAD\u2010seq) to resolve the phylogenetic history of this group and found evidence for introgression within and between clades. Our curve\u2010based approach to morphometrics of corolla symmetry demonstrated that bilateral symmetry in sect. Schistanthe is a continuous characteristic that is made up of multiple corolla components. We identified corolla symmetry types in the section and inferred at least two transitions to novel bilateral symmetry within the New Guinean radiation.The goals of our study were to investigate the phylogenetic complexity and floral diversification, with regard to flower symmetry, of sect. Rhododendron sect. Schistanthe and 17 outgroups according to a modified protocol by the QB3 Vincent J. Coates Genomics Sequencing Laboratory, University of California, Berkeley, USA, or the Genomics and Cell Characterization Core Facility, University of Oregon, Eugene, USA. Further details of the molecular methods are given in Methods We followed the method of Etter et\u2009al. for creaprocess_radtags in Stacks v.1.47 topology search in RAxML v.8.2.11 process modelling approach implemented in the R library l1ou , which has been shown to be conservative and therefore reduce the possibility of detecting unsupported shifts.Last, to examine the evolution of floral morphology during the evolutionary history of sect. Rhododendron sect. Schistanthe and outgroups using four RAD\u2010seq datasets with different levels of missing data and two reconstruction methods: ML in RAxML and the multispecies coalescent model in SVDQuartets (SVDQ). All RAxML and SVDQ reconstructions supported the monophyly of sect. Schistanthe for four clades but conflict in three clades and Quartet Differential (QD) scores. In the RAxML topology, seven clades were strongly supported : 1a, 1b, 2\u20135 and 7 and 46% (SVDQ) of nodes had QD scores <\u20090.5 as implemented in Dsuite. Based on three methods used to calculate D\u2010statistics, we found differing amounts of introgression, and here we highlight our results from DBBAA as moderate estimates of the D\u2010statistic , Euvireya C (clade 6) and Euvireya D (clade 7) . We found strong support for introgression between clades Euvireya A (clade 4) and Euvireya D (clade 7) and between clades Euvireya C (clade 6) and Euvireya D (clade 7) . We did not find strong signals of introgression with Pseudovireya (clade 1), Discovireya (clade 2), Malayovireya (clade 3) and Euvireya B (clade 5) but also lacked enough information to confidently determine this in these groups crown age of 31.04 (30.66\u201331.38) million years ago (Ma) for sect. Schistanthe Ma; its subclades, as represented by clades Euvireya A\u2013D (clades 4\u20137), had mean crown ages of 12.65 (12.50\u201312.80), 7.89 (7.76\u20138.02), 11.89 (11.74\u201312.01) and 6.33 (6.25\u20136.41) Ma, respectively ) and 166 outlines representing 97 taxa for the side view of the corolla (taxon mean n\u2009=\u20091.71 (SD\u2009=\u20090.91)) ) Fig.\u2009. The out1)) Fig.\u2009. For bot1)) Fig.\u2009. PC1 exp1)) Fig.\u2009. PC1 expK\u2010means clustering to test for distinct symmetry phenotypes in sect. Schistanthe. After evaluating 30 indices for the optimal number of clusters, nbclust used the majority rule to identify five clusters in the front view and two clusters in the side view , Discovireya (clade 2), Malayovireya (clade 3) and Euvireya B (clade 5) occupied relatively small areas of the morphospace and exhibited radially symmetric flowers that was shared by multiple species Fig.\u2009. This shum) Fig.\u2009.Schistanthe may have been ancestrally bilaterally symmetric with wider angles between the lower petal lobes than between the upper lobes. Transitions to radial symmetry and other types of bilateral symmetry occurred in the group with a novel type of bilateral symmetry arising in clade Euvireya D (clade 7).As PC1 represented depth of petal lobing but not symmetry, we used the origin of clades in PC2 and PC3 to examine symmetry evolution. Using the origin of clades in PC2, early diverging clades Pseudovireya (clade 1), Discovireya (clade 2) and Malayovireya (clade 3) appeared to have ancestral phenotypes of radially symmetric flowers Fig.\u2009. HoweverWe also examined clade occupancy of side\u2010view corolla morphospace as a means to understand flower symmetry evolution. When examining variation in PC1 and PC2, clades Pseudovireya (clade 1), Discovireya (clade 2) and Euvireya B (clade 5) occupied the smallest morphological spaces, followed by clade Malayovireya (clade 3), while clades Euvireya A (clade 4), Euvireya C (clade 6) and Euvireya D (clade 7) occupied the largest morphological spaces Fig.\u2009. When PCR.\u2009agathodaemonis, R.\u2009carringtoniae, R.\u2009goodenoughii and R.\u2009herzogii and exhibited flowers with longer, narrower corolla tubes and asymmetry in reflexing of the upper vs lower petals. The second shift was shared by R.\u2009beyerinckianum, R.\u2009calignis, R.\u2009dielsianum, R.\u2009leptanthum, R.\u2009phaeochitum and R.\u2009rarum and confirmed the origin of a second novel form of bilateral symmetry in clade Euvireya D (clade 7), which exhibited curved corolla tubes and longer upper petal lobes than lower petal lobes that were shared by multiple species Fig.\u2009. One shibes Fig.\u2009. When webes Fig.\u2009. TherefoRhododendron sect. Schistanthe and revealed the evolution of floral diversity across the tropical radiation of Rhododendron in Southeast Asia by using morphometrics of corollas to characterise variation in corolla symmetry. Corolla symmetry in sect. Schistanthe is a spectrum of bilateral symmetry that exhibits variation in asymmetry between the upper and lower parts of the corolla with respect to petal curvature, number, angle, length and/or reflexing. We suggest that the ancestral corolla phenotype for sect. Schistanthe may have been bilaterally symmetrical when taking into account both front and side views of the corolla. The New Guinean radiation of sect. Schistanthe evolved to occupy the largest corolla morphospace in the shortest amount of time and witnessed three shifts in floral evolution. This included the origin of two unique types of bilateral symmetry, in which corollas either show doubling of lateral petals and wider angles between upper vs lower petal lobes or corolla tubes become curved with longer upper than lower petal lobes.We used reduced genome\u2010wide sequencing from 114 members of Schistanthe is impeded by phylogenetic complexity and uncertainty. Prior phylogenetic studies of sect. Schistanthe based on cpDNA and the majority of clade Euvireya D (clade 7), which potentially affected the monophyly of clade Euvireya C (clade 6), and (2) a weak signal of introgression between clades Euvireya B (clade 5) and Euvireya D (clade 7), potentially affecting the position and monophyly of clade Euvireya B (clade 5). We also detected many cases of introgression within clades. Species from sect. Schistanthe hybridise easily in cultivation as long as species have similar style lengths . Our results contrast with previous work that used a more subjective coding of symmetry and only the front view of corollas, which inferred a radially symmetric corolla for sect. Schistanthe or greater than five petals. Therefore, morphometric analyses of the corolla should be expanded to the entire genus in future studies for a more holistic view of the flower. Three\u2010dimensional imaging using methods such as microcomputed tomography .Fig.\u2009S14 Morphospace variation of corollas from Rhododendron sect. Schistanthe using principal component analysis (PCA) and flower colour.Methods S1 Library preparation.Methods S2 Data processing.Methods S3 Phylogenetic analyses.Methods S4 Introgression analyses.Methods S5 Molecular dating.Methods S6 Morphometric analyses.Table\u2009S1 Samples and vouchers used in this study.Table\u2009S2 Characteristics for each dataset used in Rhododendron sect. Schistanthe analyses.New Phytologist Central Office.Please note: Wiley Blackwell are not responsible for the content or functionality of any Supporting Information supplied by the authors. Any queries should be directed to the Click here for additional data file."} +{"text": "Background: Neutrophil extracellular trap (NET) production has been implicated in the pathogenesis of thromboinflammatory conditions such as Sickle Cell Disease (SCD), contributing to heightened risk for ischemic stroke. NETs are catalyzed by the enzyme Peptidyl Arginine Deiminase 4 (PAD4) and neutrophil derived reactive oxygen species (ROS), especially NADPH oxidase (NOX) which interacts with PAD4 and is therefore critical for neutrophil function. However, the role that NOX-dependent ROS and NETs play in the accelerated cerebral microvascular thrombosis associated with thromboinflammatory conditions, such as SCD, has not been fully elucidated and is the aim of this study.Methods: The in-vitro effects of targeting PAD4 and NOX were examined using physiologically relevant NET assays with neutrophils isolated from healthy volunteers (control) and SCD patients. In addition, in-vivo intravascular effects of targeting PAD4 and NOX in the cerebral microcirculation of C57BL/6 and sickle transgenic mice (STM) were assessed using a photoactivation thrombosis model (light/dye) coupled with real-time fluorescence intravital microscopy.Results: We found that targeting PAD4 and NOX in human neutrophils significantly inhibited ionomycin dependent H3cit+ neutrophils. Targeting PAD4 and NOX in-vivo resulted in prolonged blood flow cessation in cerebrovascular arterioles as well as venules. Moreover, we were able to replicate the effects of PAD4 and NOX targeting in a clinical model of accelerated thromboinflammation by increasing blood flow cessation times in cerebral microvessels in STM. These findings concurred with the clinical setting i.e. neutrophils isolated from SCD patients, which possessed an attenuation of H3cit+ neutrophil production on targeting PAD4 and NOX.Conclusions: Taken together, our compelling data suggests that PAD4 and NOX play a significant role in neutrophil driven thromboinflammation. Targeting PAD4 and NOX limits pathological H3cit+ neutrophils, which may further explain attenuation of cerebral thrombosis. Overall, this study presents a viable pre-clinical model of prevention and management of thromboinflammatory complications such as ischemic stroke. Inflammation and thrombosis are highly intertwined processes in which systemic inflammation can beget local thrombosis, and thrombosis can amplify inflammation. Inflammatory cells, such as neutrophils, monocytes and platelets, and their pro-inflammatory mediators play an important role in thrombus formation, contributing to a \u201cthromboinflammatory\u201d phenotype -/- mice cannot citrullinate histones and are therefore incapable of forming NETs A critical step in NET formation is the decondensation of chromatin that occurs in the nucleus. Peptidylarginine deiminase (PAD4) is a nuclear enzyme that converts specific arginine residues to citrulline on histone tails, enabling chromatin decondensation and NET formation. This process is termed as citrullination Collectively, these findings highlight the crucial role that PAD4 and NETs play in thromboinflammation. Novel evidence has also suggested the potential role of PAD2 in inflammation. However, this role appears more macrophage-dependent via METosis (a process of macrophage releasing extracellular traps) and pyroptosis (caspase 1 dependent cell death) Both thrombus formation and its resolution may be regulated by ROS. ROS increases the expression and activation of tissue factor and the subsequent production of thrombin, in endothelial cells, monocytes and vascular smooth muscle cells, with ROS-generating gp91phox (NADPH oxidase [NOX]) enzymes being important contributors In-vitro experiments: vehicle 1X Phosphate buffered saline (PBS) (Life Technologies), GSK484 (10 \u03bcM. PAD4 inhibitor) Supplementary Figure 1) TM Green nucleic acid stain (Life Technologies) (1 \u03bcM) (Abcam), histone H3 mouse anti-H3Cit (1:200) were used for NET detection. In-vivo experiments vehicle , GSK484 (10 \u03bcM),Hba locus for the h\u03b1 mutation [tm1(HBA)TowHba] and homozygous at the Hbb locus for the -383 \u03b3-\u03b2A mutation [tm3TowHbb]) were purchased from Jackson Laboratory Male and female control (C57BL/6) and Sickle Cell Transgenic mice and conducted in accordance with the Declaration of Helsinki. Blood was collected from human volunteers recruited from the LSUHSC-S after obtaining consent . Blood was also obtained from SCD patients upon routine clinical visits at the Feist-Weiller Cancer Center at LSUHSC-S. All SCD patients were on chronic hydroxyurea therapy and blood was obtained just before exchange transfusion. Hydroxyurea was started at 15 mg per kilogram of body weight per day and then escalated by 5 mg per kilogram every 12 weeks until the maximum tolerated dose was achieved based on peripheral blood counts. Patients were on partial exchange transfusion every two weeks. Patients with acute infection or other chronic blood borne diseases were excluded from the study. Demographic and clinical characteristics of controls and SCD patients are included in 2o water and 10X PBS (hypotonic lysis) in 9:1 ratio to remove any contaminating erythrocytes. The solution was further centrifuged at 1000 rpm for 10 minutes. The final neutrophil containing pellet was resuspended in 1X PBS and the cells were counted by trypan blue dye exclusion using Neubauer hemocytometer. Finally, neutrophils were resuspended in DMEM with 3% fetal calf serum and kept on ice until further use. Neutrophils were routinely assessed for purity using Wright-Giemsa stain.Peripheral neutrophils were isolated from blood obtained from healthy humans and SCD patients using dextrose-histopaque separation followed by hypotonic lysis as described previously oC, 5% CO2. For immunocytochemistry, neutrophils were either stained with Sytox green (1\u00b5M) (cell-impermeable dye) or fixed , permeabilized (0.5% Triton X-100) and blocked (10% goat serum). After fixing cells were incubated with histone H3 mouse anti-H3Cit (1:200) followed by species-specific secondary antibody coupled with Alexa Fluor Dyes (Abcam). In the final step, DNA was stained with 4\u2032,6-diamidino-2-phenylindole in PBS for 10 minutes. After mounting the neutrophil containing coverslips on a glass slide, the images were visualized by Nikon Eclipse Ti inverted epifluorescence microscope (Minato-ku). In-vitro NETs were quantified by measuring the percentage of CitH3+ stained DNA over total number of neutrophils (DAPI stained) in a double blinded fashion. For precise measurements, morphological changes (characterized with protrusion of nucleus and chromatin outside the main neutrophil body) were considered for neutrophils to be positive for NETs . Pictures were taken from three to four random fields with the total of at-least 50 cells per view and the average was calculated. In addition, NETs were also quantified by analyzing Sytox green intensity by plate reader (Synergy H1).Neutrophils were seeded on poly-l-lysine coated coverslips (Discovery Labware) in a 24-well plate or transferred directly to a 96-well plate and were stimulated for 3 hours at 372) for 2 hours. DHR123 (25 \u03bcM) was added to the wells for 15 minutes before fluorescence reading using a plate reader or ionomycin stimulation and were kept in a humidified incubator .Anesthetized mice were kept under the microscope after jugular vein cannulation and open window craniotomy. Thrombosis in cerebral vessels was induced using the light/dye thrombosis model p value. Data that passed the normality assumption was analyzed using Student's t-test (two groups) or ANOVA with Bonferroni post-tests (more than two groups). Data that failed the normality assumption were analyzed using the non-parametric Mann-Whitney U test (two groups) or Kruskal-Wallis with Dunn's test (more than two groups). Analysis was performed using Graph Pad Prism 9 software (San Diego). Data are shown as mean values \u00b1 standard error of the mean (SEM). Differences were considered statistically significant at a value of p<0.05.All data was tested to follow a normal distribution using Kolmogorov-Smirnov test of normality with Dallal-Wilkinson-Lillie D'Agostino-Pearson omnibus normality test for corrected For original data, please contact felicity.gavins@brunel.ac.uk.Figure , n \u2265 5). Sytox green intensity was used for expression of neutrophil extracellular DNA Figure (n \u2265 5) shows that GSK484 significantly inhibits ionomycin-stimulated extracellular DNA production (p<0.001), an effect not shared by neutrophils treated with VAS3947. Furthermore, upon co-administration of GSK484+VAS3947, the PAD4 inhibitor still retained its inhibitory effects , inferring NOX dependent ROS inhibition is not necessary for extracellular DNA production, suggesting neutrophil extracellular DNA production is NOX independent.Having previously discovered that neutrophils, and more importantly their NETs, play a key role in cerebral thrombosis +. Figure , n \u2265 5) was used + is the most common NET biomarker that has been associated with experimental thrombosis Figure (n \u2265 5) shows that GSK484 (p<0.0001) when used alone or in combination with VAS3947 significantly reduced the percentage of ionomycin-stimulated neutrophils that were positive for H3Cit+ (p<0.0001), suggesting that targeting either PAD4 or NOX dependent ROS production has significant effect on histone hypercitrullination. Interestingly, similar to the effects observed with GSK484, VAS3947 was also able reduce the percentage of H3Cit+neutrophils , an effect which was opposite to that observed with extracellular DNA production . These results suggested that targeting both PAD4 and NOX results in suppression of ionomycin-stimulated H3Cit+ neutrophils. The above differences clearly suggest differences in extracellular DNA production and H3cit+ neutrophils production and targeting To specifically characterise pathological NETosis specific NET immunucytological statin Citrullinated histone-3 as well as H3Cit+ neutrophils . In a similar fashion VAS3947 was unable to suppress PMA dependent extracellular DNA production further suggesting that extracellular DNA production is NOX independent . However, VAS3947 significantly suppressed PMA dependent H3Cit+ neutrophils (p<0.001) . Additionally, VAS3947 was able to inhibit PMA dependent ROS production (p<0.001) . On the contrary, ionomycin was not able to increase ROS production in human neutrophils . Finally, there was no effect of ROS production in VAS3947 treated ionomycin stimulated human neutrophils . These findings further confirm the importance of ROS in H3Cit+ neutrophil production irrespective of neutrophil stimuli.It is known that different NET stimuli may have variable responses depending on the NET-associated intracellular signalling pathway engagement + neutrophils, we wanted to see the effect of such approach on vascular thrombosis using fluorescence intravital microscopy with light/dye injury model of thrombosis Figure (n \u2265 5) shows that both GSK484 and VAS3947, when used alone or in combination, were able to significantly prolong blood flow cessation in cerebrovascular arterioles as well as venules clearly demonstrating that targeting PAD4 and NOX production is an effective strategy to mitigate vascular thrombosis.Having found that inhibition of PAD4 and ROS hold significant repressive effects on extracellular NET production as well as H3CitFigure 4) and wanted to see whether the perturbation of PAD4 and NOX will result in attenuation of thromboinflammatory signalling. VAS3947 failed to inhibit ionomycin-induced extracellular DNA production but sufficiently inhibited ionomycin-induced histone citrullination in SCD neutrophils . GSK484 failed to inhibit ionomycin-induced extracellular DNA production but was able to significantly attenuate ionomycin-induced histone citrullination in SCD neutrophils . Combination of GSK484 and VAS3947 resulted in significant reduction of both ionomycin dependent histone citrullination as well as extracellular DNA production . However, GSK484 failed to inhibit PMA dependent extracellular DNA production and histone citrullination in SCD neutrophils . Finally, VAS3947 resulted in significant reduction of both extracellular DNA production and PMA-dependent histone citrullinetion . All the above results further implicate PAD4 and ROS as an important target for inhibition of pathological NET production.To build on our murine findings and translate them to a clinical setting, we next isolated neutrophils from SCD patients . Inhibition of PAD4 and NOX significantly increased blood flow cessation times in cerebral microvessels (arterioles as well as venules) in STM, hence replicating the effects seen in C57BL/6 mice .Finally, to further validate our findings from SCD neutrophils, we used a humanised transgenic mouse model of SCD coupled to intravital microscopy to study thrombosis in a pathophysiological setting attenuates murine cerebrovascular inflammation and modifies neutrophil behaviour in human neutrophils, including SCD, towards an anti-thromboinflammatory phenotype. GSK 484 fails to inhibit PMA dependent extracellular DNA and H3Citin-vitro system by treating neutrophils with various stimuli such as ionomycin and PMA which can significantly promote pro-inflammatory phenotype Neutrophils are implicated in the pathogenesis of various inflammatory conditions including cardiovascular diseases + neutrophil production, it failed to suppress PMA dependent H3Cit+ neutrophil production. These results suggest that PMA stimulation either bypasses PAD4 activation, or it employs an accessory ROS dependent PAD4 independent pathway for H3Cit+ neutrophil production + neutrophil production after ROS targeting in ionomycin as well as PMA stimulated neutrophils. Ionomycin also resulted in minimal ROS production, which is likely explained by the fact that ionomycin causes NET production by engaging PAD4 complex, independent of NOX It is known that one of the key elements of pathological NET production is histone citrullination PAD4 activation and translocation is a critical step for NET production and dysregulated PAD4 activation is known to enhance inflammatory responses in various chronic disease such as rheumatoid arthritis and diabetes mellitus SCD provides an excellent disease paradigm to study accelerated thrombosis. PAD4 as well as NOX are known to provoke thrombosis in known clinical models of inflammation potential therapeutic strategy and target(s) for drug discovery programs focussed on the treatment and management of thrombosis and inflammation, conditions which underpin many of today's global health challenges.Our discovery that PAD4 and NOX-dependent ROS modifies neutrophil behaviour in thromboinflammatory settings provides Supplementary figures and table, video legends.Click here for additional data file.Supplementary video 1.Click here for additional data file.Supplementary video 2.Click here for additional data file.Supplementary video 3.Click here for additional data file.Supplementary video 4.Click here for additional data file."} +{"text": "Streptococcus pneumoniae and Legionella pneumophila is only recommended in patients with severe pneumonia according to the 2019 guidelines from the American Thoracic Society (ATS) and Infectious Diseases Society of America (IDSA). Unnecessary testing for these organisms may increase antibiotic utilization. This quality improvement project evaluated whether patients who received a Streptococcus pneumoniae or Legionella pneumophila urine antigen met criteria for testing.Urinary antigen testing for Streptococcus pneumoniae or Legionella pneumophila within 3 days of admission to the Veterans Affairs Connecticut Healthcare System between September 1, 2018 and September 1, 2021. We determined whether patients had clinically defined pneumonia per National Healthcare Safety Network criteria. Among patients that met criteria for clinically defined pneumonia, the subset with severe pneumonia according to the ATS/IDSA guidelines was identified. We used the ATS/IDSA guidelines to determine whether patients received guideline-concordant choice of therapy.We conducted a cohort study of patients who received a urinary antigen test for Streptococcus pneumoniae antigen test and 1 patient had a positive test for Legionella. Among patients with clinically defined pneumonia, 74% (n=75) received guideline-concordant choice of therapy and 74% (n=75) received an antibiotic duration >5 daysWe identified 352 patients who received a urinary antigen test within 3 days of admission. Mean age was 72.6 years, 97% (n=278) were male sex, and 86% (n=301) were White race. Common comorbidities included heart disease , lung disease , and diabetes . Overall, 36% (n=127) of patients met criteria for clinically defined pneumonia, and 7% (n=25) met criteria for severe pneumonia. 9 patients had a positive Diagnostic and Therapeutic Characteristics of Veterans According to Presence of PneumoniaStreptococcus pneumoniae or Legionella pneumophila met criteria for testing per ATS/IDSA guidelines, and length of therapy was prolonged in the vast majority of patients. These data support the need for a clinical decision-making tool to reduce unnecessary testing and treatment for pneumonia at our institution.Only 7% of patients who underwent urine antigen testing for All Authors: No reported disclosures."} +{"text": "Irritable bowel syndrome (IBS) is characterized by chronic abdominal pain. Previous work has shown IBS fecal supernatants (FS) increase the excitability of nociceptive neurons compared to healthy volunteer FS. Altered production of intestinal mediators, including elevated histamine content and proteolytic activity, have been implicated in these effects. We hypothesized that the effects of histamine and proteases on neuronal excitability may be synergistic.1) Determine whether the excitatory effect of IBS patient FS can be abolished by histamine receptor antagonists and protease inhibitors. 2) Investigate whether the excitatory effects of histamine and proteases potentiate each other\u2019s effects on visceral nociceptionExtracellular recordings were performed, measuring action potentials from mechanosensitive extrinsic afferent nerves innervating the mouse colon during colonic distensions to 60 mmHg. Colons were perfused (20 minutes) with histamine receptor antagonists or protease inhibitors prior to treatment with the IBS patient FS. Perforated patch clamp experiments were used to measure neuronal excitability from mouse dorsal root ganglion neurons by recording rheobase . This technique was used to examine the effect of subthreshold concentrations of histamine (1\u00b5M) and trypsin (5nM), as well as their simultaneous administration on the neurons.Luminal administration of histamine antagonists or protease inhibitors blocked the excitatory effect of the IBS patient FS . Patch clamp experiments revealed that a subthreshold concentration of either histamine (1\u00b5M) or trypsin (5nM) had no effect on neuronal rheobase, whereas the combination significantly decreased neuronal rheobase .These findings demonstrate that either histamine receptor antagonists or protease inhibitors inhibit the excitatory effect of IBS patient FS and suggest that a potentiating effect exists between the actions of histamine and proteases on nociceptive neurons.None Declared"} +{"text": "Unhealthy behaviours are associated with increased sickness absence (SA), but few studies have explored these associations using person-oriented approach. We aimed to identify latent classes of unhealthy behaviours among female and male employees and examined their associations with subsequent SA.Health behaviours were derived from the Helsinki Health Study questionnaire survey, collected in 2017 among 19-39-year-old employees of the City of Helsinki, Finland. The questionnaire data were linked to employer's SA register. Latent class analysis was used to identify underlying profiles of unhealthy behaviours and negative binomial regression was used to examine their associations with subsequent SA among 3228 women and 771 men. The mean follow-up time was 2.1 years.Among women, we identified 3 latent classes: 1) healthy behaviours (81% of women), 2) binge drinking and tobacco use (12%), and 3) inadequate F&V consumption and insufficient sleep (7%). Classes 2 and 3 showed increased rates for subsequent SA compared to class 1, regardless of the length of SA spells . Among men, we identified 3 latent classes: 1) healthy behaviours (51% of men), 2) binge drinking and tobacco use (19%), and 3) inadequate F&V consumption, binge drinking and tobacco use (30%). While classes 1 and 2 were not different in terms of subsequent SA, class 3 had increased rates of subsequent, particularly short-term SA .Preventive actions should consider simultaneously several unhealthy behaviours while aiming to reduce employees\u2019 SA. These actions might benefit from regarding potential gender differences in the clustering of unhealthy behaviours and their associations with SA.\u2022\u2002Preventive actions to reduce sickness absence should consider clustering of unhealthy behaviours among employees.\u2022\u2002Potential gender differences need to be regarded in these actions."} +{"text": "Impaired airway protection was also an independent predictor for longer tracheostomy tube duration . The majority of our study patients presented with complex laryngeal findings which were associated with impaired airway protection. We suggest a proactive standardized scoring and review protocol to manage this complex group of patients in order to maximize health outcomes and ICU resources. Early laryngeal assessment may facilitate weaning from invasive mechanical ventilation and liberation from tracheostomy, as well as practical and objective risk stratification for patients regarding decannulation and feeding.To explore laryngeal function of tracheostomised patients with COVID-19 in the acute phase, to identify ways teams may facilitate and expedite tracheostomy weaning and rehabilitation of upper airway function. Consecutive tracheostomised patients underwent laryngeal examination during mechanical ventilation weaning. Primary outcomes included prevalence of upper aerodigestive oedema and airway protection during swallow, tracheostomy duration, ICU frailty scores, and oral intake type. Analyses included bivariate associations and exploratory multivariable regressions. 48 consecutive patients who underwent tracheostomy insertion as part of their respiratory wean following invasive ventilation in a single UK tertiary hospital were included. 21 (43.8%) had impaired airway protection on swallow (PAS\u2009\u2265\u20093) with 32 (66.7%) having marked airway oedema in at least one laryngeal area. Impaired airway protection was associated with longer total artificial airway duration ( Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) results in a high incidence of respiratory distress. Between 1/9/20\u201310/5/21, 7644 individuals in the UK received invasive ventilation within 24\u00a0h of admission to hospital [Tracheostomy decannulation timing and approaches are numerous . A key qProspective data collection was used to gather information on acute laryngeal anatomy and physiology following invasive ventilation and tracheostomy tube placement for treatment of COVID-19. Nasendoscopic examination before decannulation is standard practice in our institution. There is no control group in this study, as the evaluation was undertaken during the second peak of Covid-19 in the UK, where there was no available comparable group due to the intense resource utilisation for COVID-19 patients. We seek to describe this specific pathophysiology, to inform other ENT, intensive care and Speech and Language Therapy teams and to improve outcomes for this complex group of patients. Future prospective work is underway to facilitate comparison between case\u2013control with appropriate methodology and study design. This work uses data provided by patients and collected by the NHS as part of their care and support at the Queen Elizabeth Hospital Birmingham NHS Foundation Trust. It has been approved by University Hospitals Birmingham NHS Foundation Trust, Clinical Audit Registration & Management System and the COVID-19 research facilitation group under application reference CARMS-17155.From January 1 to April 28, 2021, we included all consecutive ICU patients considered for tracheostomy weaning at the Queen Elizabeth Hospital Birmingham. Included were those with severe respiratory failure secondary to SARS-CoV-2 with positivity confirmed by real-time polymerase chain reaction testing or non-directed bronchial lavage/aspirate. Once the patient was able to tolerate oxygen delivery via tracheostomy mask (without invasive ventilation), they were eligible for a fiberoptic nasendoscopy.We have described percutaneous tracheostomy tube insertion methods and operational aspects of our MDT previously . In brieAn ENT surgeon and SLT completed the laryngeal assessment via nasendoscopy when the patient tolerated oxygen supplementation via a tracheostomy mask without invasive ventilation. Where patients were sufficiently alert, a swallow assessment was completed with and diet fluid recommendations. The examination was recorded on the AMBU disposable scope system (Ambu\u00ae aScope\u2122 4 RhinoLaryngo Slim). We used a predefined proforma, including laryngeal and pharyngeal motor and sensory assessment and standardised oedema and airway protection scoring during swallowing. The decision to decannulate was made by the treating team (intensive care or respiratory) using their usual clinical parameters. Data collection for audit purposes ceased after decannulation.The revised Patterson oedema scale and PenePresence of Intensive Care acquired weakness (ICUAW) was assessed by a physiotherapist at ICU discharge using Medical Research Council (MRC) sum score . A validt-tests for continuous variables and Pearson\u2019s \u03c72 test, likelihood ratio or Fisher\u2019s exact test for proportions as appropriate. Post-hoc, we conducted two exploratory main effect regression analyses for two study outcomes: a logistic backward stepwise regression [p\u2009<\u20090.05.For analyses, we dichotomized PAS describing airway protection as \u2018normal\u2019 (PAS scores of either 1 or 2) or \u2018impaired\u2019 (PAS scores of 3 and above). All continuous variables were summarized according to mean with standard deviation (SD) and median with interquartile range (IQR) depending on whether data were normally distributed, and ordinal/categorical data summarized according to frequency counts. Following stratification of the sample according to normal or impaired PAS, bivariate comparisons were conducted using Mann\u2013Whitney or unpaired 2-sided gression to explogression . SignifiOf 146 consecutive patients, 48 met inclusion criteria during our study period. Mean age (SD) was 56.7 (10.7) years and 31 (64.6%) were male. On admission, the majority had a Charlson comorbidity Index of 3 or above and 9 (18.8%) were living with mild pre-existing frailty before admission. Forty-one (85.4%) were without underlying chronic respiratory diseases . Other demographics and baseline characteristics are available in Table Respiratory and physical characteristics were reported/measured during the patients\u2019 stay in the ICU and upon ICU discharge and transfer to the ward Table . Across p\u2009=\u20090.006) and 38 patients respectively. Those with and without normal airway protection did not differ significantly in regards to demographic and baseline variables except for BMI (p\u2009=\u20090.04) and asthma (p\u2009=\u20090.003). Those with normal airway protection had greater BMI and very few patients in our sample had underlying asthma. Impaired airway protection (as determined on PAS) was associated with longer total duration of artificial airway (p\u2009=\u20090.008), longer tracheostomy tube duration (p\u2009=\u20090.007), and multiple intubations (p\u2009=\u20090.006). Airway protection impairment was also significantly different in those with greater critical care weakness upon ICU discharge was observed in 21 (43.8%). At the time of nasendoscopy, impaired airway protection was significantly associated with an inability to swallow fluids and solids safely for 29 . For supraglottic components (region 2), normal airway protection was associated with lower oedema scores for the valleculae (p\u2009=\u20090.01). Those with normal airway protection had significantly better management of their secretions (p\u2009=\u2009\u2009<\u20090.001). Individual oedema scale components and secretion scores are presented in Table p\u2009=\u20090.003) and food (p\u2009=\u20090.006). Following our exploratory regression (Table p\u2009=\u20090.02). One patient presented with a vocal cord praxis which recovered within 30\u00a0days and didn\u2019t preclude decannulation, another presented with a vocal cord palsy and required targeted SLT rehab before the tracheostomy was also removed successfully. No granuloma or stenosis were identified.We observed marked airway oedema with 32 (66.7%) patients scoring 3 or above on at least one revised Patterson oedema Scale component. Oedema scores\u2009\u2265\u20093 were present in at least one anatomical area for 29 60.4%) patients in region 1, and for 22 (45.8%) in region 2. When considering the individual glottic components (region 1) and airway protection, lower oedema scores favored normal airway protection for false vocal fold movement/oedema (% patientp\u2009=\u20090.02). Upon discharge from ICU, 19 (39.6%) had severe to very severe ICU acquired weakness with 3 (6.3%) able to mobilize\u2009<\u200930\u00a0m. Critical care acquired weakness on ICU discharge was significantly different between groups with impaired airway protection (p\u2009=\u20090.04); impaired airway protection was also an independent predictor for tracheostomy tube duration . Mean (SD) ward length of stay was 27.7 (19.2) days with an overall hospitalization duration of 71.4 (30.4) days. At hospital discharge, the majority 31(64.6%) tolerated regular fluids and solids (with only one patient remaining nil by mouth). There were no significant differences between those with and without normal airway protection and physical rehabilitation outcomes at discharge. Furthermore, overall hospitalization durations were not significantly different between those with and without normal airway protection. Details regarding physical rehabilitation outcomes are available in Table Mean (SD) length of stay in the ICU was 43.7 21.6) days. Those with impaired airway protection had significantly longer mean (SD) length of ICU stay (days) when compared to those with normal airway protection: (53.6 (27.2) versus 36.6 (12.6); days. ThSince the emergence of COVID-19, publications exploring tracheostomy pathways , 19 havePatients with impaired airway protection had significantly more complex respiratory recovery specifically longer durations of artificial airways and more frequent intubations. In addition, the majority required downsizing or fenestrated tubes. Although our analyses were exploratory, we suggest impaired airway protection may be an independent predictor of tracheostomy duration. While this aligns with other studies on critically ill patients without COVID-19 , this isA common finding in survivors of critical illness is ICUAW . The majThis was a quality assurance audit, undertaken during the third COVID-19 surge in the UK and as a result, our study had limitations and should be considered through the lens of the following design caveats. Our small sample size without a control group limits generalizability and does not elucidate the potential differences between this populations as compared to those with critical illness without COVID-19. However, it is pertinent to couch this methodological limitation in line with other pandemic publications without control groups, which have contributed fundamental learning to this novel and emergent pathophysiology and clinical presentation . In addiOur findings highlight the functional relationships between the anatomy and physiology of the larynx and cumulative outcomes following artificial airway insertion. In our institution, patients who required tracheostomy following COVID-19 presented with impaired airway protection and marked airway oedema in at least one laryngeal area. Impaired airway protection was associated with longer total duration of artificial airway, longer tracheostomy tube duration, and multiple intubations. We suggest proactive assessment, standardised scoring, and patient risk stratification to enable the clinical team to create collaborative and effective decannulation plans."} +{"text": "Repeat blood cultures (BCxs) beyond 48 hours are often obtained despite negative initial BCxs in hospitalized pediatric hematology/oncology patients. This study seeks to determine the yield of repeat BCxs after negative initial cultures in these patients and to characterize new positive BCxs beyond 48 hours and the clinical contexts in which they were obtained.A retrospective review utilizing MedMined Inc. Data Mining Surveillance database was conducted on all BCxs obtained on hospitalized patients on the pediatric hematology/oncology unit at Riley Hospital for Children in Indianapolis, IN from January 2015 to February 2021. Exclusion criteria are shown in Fig.\u00a01. Patient episodes in which a new pathogen was identified on a repeat BCx more than 48 hours after negative initial BCxs were further investigated via electronic medical record review.A total of 1,362 BCx sets were obtained beyond 48 hours in 792 patient hospitalizations, resulting in 303 positive BCxs . Of these positive cultures, 193 were the same pathogen cultured on day 0 and 74 were contaminant cultures (in 4.0% (23/573) of patient hospitalizations without a positive BCx before 48 hours). Only 36 (2.6%) of positive BCxs beyond 48 hours were determined to be new pathogens, or commensals treated as pathogens, that were not cultured before 48 hours, corresponding to seven patient hospitalizations (1.2% (7/573) of patient hospitalizations without a positive BCx before 48 hours). The majority (6/7) of these patients were neutropenic and on broad spectrum antibiotics when the new positive BCxs were obtained. Fever pattern was prolonged in one patient and recurrent in six. No deaths occurred in these seven patients. All patients with new, true pathogens on BCxs beyond 48 hours (n=5) were either hemodynamically unstable (n=3) or had clinical changes the day the new positive BCx was drawn.The yield of repeat BCxs beyond 48 hours in hospitalized pediatric hematology/oncology patients with negative initial BCxs is low, while the associated costs are high. Repeat BCxs beyond 48 hours after negative initial cultures need not be obtained in febrile patients that remain hemodynamically stable and without clinical changes.Cassandra S. Prather, MD, Eli Lilly and Company: Spouse is employed by Eli Lilly and Company. Spouse owns stock in the company.|Eli Lilly and Company: Stocks/Bonds."} +{"text": "Evaluate the type and incidence of postoperative complications after surgery for deep infiltrative endometriosis at Biocor Hospital.th postoperative day were recorded.Our observational study involved a multidisciplinary surgical team that performed laparoscopy on 154 patients suffering from pelvic pain. Surgical complications occurring up to the 30Mean age patient age was 34.1 years. Infertility was present in 69 (45%) although 31% had not attempted to get pregnant. Dysmenorrhea was the most frequent symptom (79.3%) followed by chronic pelvic pain (59.7%) and deep dyspareunia . Most cases required extensive surgery as the majority were classified as severe endometriosis (ASRM grade IV). The most frequent surgical procedures were: 136 adhesiolysis, 100 intestinal surgeries (85 retosigmoidectomies), 92 peritonal lesion excision, 39 vaginal resections, 19 myomectomies, 21 hysterectomies and 5 partial bladder resections. Postoperative complications were recorded in 14 (9.59%) patients: 8 (5.48%) major complications and 6 (4.11%) minor. Major complications included blood transfusion (n=2) retosigmoid anastomosis dehiscence (1), rectovaginal fistula (n=1), urinary fistula (n=1), deep vein thrombosis (n=1), lower limb compartment syndrome with motor deficit (n=1) and one intestinal obstruction (n=1). Minor complications were abdominal wall infection (n=3), peripheral neuropathy (n=3), bladder atony (n=1) and bladder perforation (n=1). No deaths were observed. All major complication cases underwent retosigmoidectomy associated with vaginal resection (n=6), uterosacral ligament excision (n=5) or hysterectomy (n=3).The surgical treatment of DIE is complex and subject to complications. The surgical expertise of a multidisciplinary team plays a vital role in this setting. Deep infiltrative endometriosis (DIE) has been defined as endometriosis that penetrates more than 5 mm under the peritoneal surface . EstabliLaparoscopic eradication of all visible implants is feasible and safe when surgery is performed in a referral center and coloproctologist (LMPC). Only cases with histological confirmation of endometriosis were included. Relevant preoperative, intraoperative, and postoperative data were retrieved and recorded in an Excel spreadsheet. Surgical complications occurring up to the 30All women underwent gynaecological examination, pelvic transvaginal and abdominal ultrasonography in order to evaluate the presence of pelvic endometriosis before surgery. Other diagnostic tests were performed when indicated, as previously described rectosigmoid anastomosis dehiscence (1), rectovaginal fistula (n=1), urinary fistula (n=1), deep vein thrombosis (n=1), lower limb compartment syndrome with motor deficit (n=1) and one intestinal obstruction (n=1). Minor complications were abdominal wall infection (n=3), peripheral neuropathy (n=3), bladder atony (n=1) and bladder perforation (n=1). No deaths were observed. All major complication cases underwent retosigmoidectomy associated with vaginal resection (n=6), uterosacral ligament excision (n=5) or hysterectomy (n=3).We report the results of 154 surgeries for DIE performed by our multidisciplinary surgical team which is composed of three gynecological surgeons, one coloproctologist and an urologist as well as reproductive endocrinology and infertility specialist. Surgical management of severe disease often requires a multidisciplinary team approach, because surgery may be complex, requiring input from colorectal and urologic colleagues. In our center we have been practicing complete surgical excision of all affected tissue for more than a decade. Our current multidisciplinary approach to surgery was completely established by the beginning of 2000. Laparoscopic skills of the gynecologists, colorectal surgeons, and urologists involved in the endometriosis team were fully developed by this time. The aim of this study was to examine the short term surgical outcome in this group of patients in terms of the rate of surgical complications.We decided to include only laparoscopies performed by the same gynecologist (IA) and coloproctologist (LMPC) in order to eliminate possible confounding factors although the other surgeons usually participate actively in all procedures.The 154 women operated on by our team suffered from pelvic pain from but 45% were also infertile and 31% had not even attempted to get pregnant. They were also young (mean age 34.1 years) so fertility preservation was an important issue for many of them. It is now accepted that effective symptomatic management of DIE requires complete excision of the rectovaginal and intestinal disease were classified as severe endometriosis (ASRM grade IV). This reflects the establishment of our endometriosis team over the years with increasing numbers of women being referred to us for specialized treatment as well as the reported delay of surgical diagnosis of deep infiltrating endometriosis which is significantly longer for patients with advanced stage IV disease . Our rate is compatible with other published studies and anastomosis dehiscence (1%). After a mean follow-up of 14 months that included 42 patients, recurrence of clinical symptoms (pelvic pain and dyspareunia) was observed in 8 cases as well as 4 cases of asymptomatic intestinal wall endometriosis recurrence which was identified by ultrasonography (Networks and centres of excellence within a multi-disciplinary context is the only way forward to ensure that women with persistent/chronic endometriosis receive consistent, evidence-based and cost-efficient care within a framework which is able to provide excellence, continuity of care, a multidisciplinary approach, research, training and cost effective management (Our findings are in agreement with recent published papers which emphasize that a well-trained multidisciplinary team can perform surgical treatment of DIE laparoscopically with low incidence of major complications (DIE affects young women for whom fertility preservation is a major issue. The surgical treatment of DIE is complex and subject to complications. This should be taken into consideration when deciding on treatment. Laparoscopic excision of deeply infiltrating pelvic endometriosis within a multidisciplinary setup in a tertiary referral center appears to be safe with a low rate of significant short-term complications. The surgical expertise of a multidisciplinary team to allow for a fertility-sparing approach plays a vital role in this setting since most women we treated were infertile, had ovarian endometriomas and a third had never attempted pregnancy."} +{"text": "Workplace violence (WPV) is a worldwide health problem with major individual and societal consequences. Previously identified predictors of WPV include working in psychiatry and work stress.To investigate WPV trends during Norwegian doctors\u2019 careers and assess individual long-term predictors in a longitudinal study.Two nationwide medical student cohorts (n=1052) who graduated 6 years apart were surveyed at graduation and 4 (T2), 10 (T3), 15 (T4) and 20 (T5) years after graduation . WPV was measured as multiple threats or acts of violence experienced at least twice. Individual predictors were obtained at T1 and work-related factors at T2\u2013T5. WPV was analysed using repeated measures .The prevalence of multiple threats and acts of violence declined at T2\u2013T5 (p<0.001). Adjusted predictors of threats were male gender , vulnerability traits , older cohort and working in psychiatry . Adjusted predictors of acts were male gender , older cohort and working in psychiatry .Higher rates of multiple threats and acts of violence were observed during early medical careers, with men at higher risk. Low levels of vulnerability traits (neuroticism) predicted independently the experience of violent threats. A cohort effect indicated a reduction in WPV (both threats and acts) in the younger cohort."} +{"text": "Automatic pathology diagnosis using WSI gradually becomes a research hotspot in biomedical imaging domain Low level WSI loading. 2) Color space conversion from RGB to gray. 3) Inverse binarization to generate binary image. 4) Hole filling of the binary image. 5) Small object removal. 6) Contour finding.The proposed WSI tissue localization is very efficient as it is entirely based on basic image processing techniques and applied on low level image, which could act as a preprocessing step for automatic WSI analysis. Researchers can focus their analysis on those patches inside the located tissue regions and avoid those irrelevant background regions."} +{"text": "Borderline personality manifests in female adolescence and youth by higher frequency of deviant behaviors and suicidal ideations. Psychological models suggests that both perception and relationship to physical pain as well as psychological pain could increase the risk.This study concentrates on the relationship between relationship to physical and psychological pain and reported deviant behavior in female adolescents.204 female adolescents (13-21 years old) filled checklist appraising alcohol use, drug use, aggressive behavior, suicidal ideations and emotional difficulties , Interpersonal Needs Questionnaire , Discomfort Intolerance Scale , The Pain Catastrophizing Scale .Elder females more frequently reported substance use (r=.23-.28) and less frequently aggressive behavior (r=-.19) while suicidal ideations were unrelated to age. Females reporting higher perceived burdensomeness and emotional difficulties also reported higher alcohol use (r=.25-.29), aggressive behavior (r=.37-.42) and suicidal ideations (r=.64-.84). Thwarted belongingness correlated with suicidal ideations (r=.50) and aggressive behavior (r=.26). Higher alcohol use was associated with catastrophizing of pain in the form of magnification and helplessness (r=.17) while suicidal ideations and aggressive behavior were related to ruminations, magnification and helplessness (r=.23-.33). Only correlations between aggression and pain catastrophizing remained significant after statistical control of psychological pain (r=.15-.22).After control for psychological pain, only aggressive behavior is related to catastrophizing of physical pain. Study is supported by Russian science Foundation, project 22-28-01524.Study is supported by Russian Science Foundation, project 22-28-01524."} +{"text": "The coronavirus pandemic has led to an exceptional number of critical care hospitalizations followed by extended recovery periods that necessitate familial support. Using a qualitative descriptive approach, this study aimed to examine the strategies used by families to adjust to the caregiving role. Semi-structured interviews of patients who had been recently discharged from the Intensive Care Unit (ICU) (n=16) along with their family caregivers (n=16) were thematically analyzed. Three major themes were identified that highlight how family caregivers adapt to the caregiving role following an ICU COVID-19 related hospitalization including 1) engaging the support of family and friends, 2) shifting responsibilities to accommodate caregiving, and 3) managing one\u2019s emotions. Additional themes more specifically related to managing COVID-19 care included: 1) managing infection control, 2) care recipient\u2019s need for independence, and 3) managing support services. Flexibility and sufficient support allowed family caregivers to manage their new caregiving role and function optimally."} +{"text": "Incorporation of 13C into low GC mol% AOA and virus genomes increased DNA buoyant density in CsCl gradients but resulted in co-migration with dominant non-enriched high GC mol% genomes, reducing sequencing depth and contig assembly. We therefore developed a hybrid approach where AOA and virus genomes were assembled from low buoyant density DNA with subsequent mapping of 13C isotopically enriched high buoyant density DNA reads to identify activity of AOA. Metagenome-assembled genomes were different between the two soils and represented a broad diversity of active populations. Sixty-four AOA-infecting viral operational taxonomic units (vOTUs) were identified with no clear relatedness to previously characterised prokaryote viruses. These vOTUs were also distinct between soils, with 42% enriched in 13C derived from hosts. The majority were predicted as capable of lysogeny and auxiliary metabolic genes included an AOA-specific multicopper oxidase suggesting infection may augment copper uptake essential for central metabolic functioning. These findings indicate virus infection of AOA may be a frequent process during nitrification with potential to influence host physiology and activity.Ammonia-oxidising archaea (AOA) are a ubiquitous component of microbial communities and dominate the first stage of nitrification in some soils. While we are beginning to understand soil virus dynamics, we have no knowledge of the composition or activity of those infecting nitrifiers or their potential to influence processes. This study aimed to characterise viruses having infected autotrophic AOA in two nitrifying soils of contrasting pH by following transfer of assimilated CO It is also responsible for major losses of applied fertiliser N in soil, generating atmospheric pollution via direct and indirect production of nitrous oxide ) to stimulate AOA growth and potential enrichment of AOA-infecting viruses. Microcosms were incubated at 25\u2009oC for 30 days was then pooled for each replicate microcosm and metagenomes sequenced co-migrated within the CsCl gradient with more abundant community 12C genomes with higher GC mol% (>57%), reducing AOA host and virus sequencing depth, contig assembly and binning. Consequently, only one medium-quality AOA metagenome-assembled genome (MAG) was recovered from HBD DNA and no AOA-associated virus contigs were identified amongst 290 viral operational taxonomic units (vOTUs) \u226510\u2009kb representing both dormant and active populations. Individual sequence reads from HBD DNA libraries from both 12C- and 13C-incubations were then mapped to identify activity with a significantly greater relative abundance in 13C-incubated samples demonstrating isotope incorporation. For taxonomically defined contigs of hosts or host-linked viruses, in comparison to HBD, assemblies from LBD libraries increased the relative proportion of contigs from the AOA-containing Thermoproteota phylum and Thermoproteota-infecting virus contigs from 0.3 to 2.8% and 0 to 22.3%, respectively Nitrobacter MAG (MAG 15) to NO3- during the second step of canonical nitrification. Mapping of HBD sequence reads revealed that eight of the ten MAGs were derived from active autotrophic populations from pH 4.5 soil and three Nitrososphaera genus populations from pH 7.5 soil. No evidence of autotrophic growth was observed for populations represented by a further two MAGs within the Nitrososphaerales. Both are from genera with no cultivated representatives with MAG 112 from genus TA-21 (amoA-gene lineage NS-\u03b2-2 [amoA-gene lineage NS-\u03b4-2.1) representing a novel genus (designated here as CS63). The latter belongs to the amoA gene-defined NS-\u03b4 lineage that while widely found in soil, is rarely implicated in contributing to ammonia oxidation. Although these two MAGs may simply be from AOA populations not active under the incubation conditions used, they are also consistent with microorganisms possessing physiologies distinct from cultivated AOA.To facilitate analysis of low GC mol% genomes while also identifying activity, we developed a hybrid approach combining metagenomic analysis of low buoyant density (LBD) 15) Fig.\u00a0, contribons Fig.\u00a0. Consiste NS-\u03b2-2 ) and MAG12C and 13C libraries revealed that 27 contigs from both soils (42%) were enriched in 13C and thus confirming infection during incubation and transfer of C from AOA hosts to viruses.After predicting contigs of virus origin using established bioinformatic tools, those from AOA-infecting viruses were identified with a custom database of hallmark genes from AOA proviruses and analysis of other shared homologues see\u00a0. Specifi13C incorporation into virus genomes, it is potentially limited in facilitating confident bioinformatic prediction of free temperate virus particles versus proviruses present from a previous infection of a currently active host. While the integration status of a temperate virus can be predicted bioinformatically, using targeted \u201cviromes\u201d metagenomes of filtered virus particles will provide greater confidence of identifying genomes from free virus particles.Soil AOA viruses were distinct from characterised archaeal and bacterial viruses. Gene-sharing network analysis using vConTACT v2.0 did not \u22125) sub-unit gene sequences and were similar to those recovered from other predicted AOA virus or provirus genomes only Fig.\u00a0. In addi\u22125) Fig.\u00a0. Copper Nitrobacter MAG was obtained and both AOB and NOB viruses were also predicted, the hybrid approach developed in this study targeted low GC mol% DNA and was specifically tailored for the analysis of AOA populations and their viruses. A broader analysis of all viruses infecting nitrifying prokaryotic populations would therefore likely require a more comprehensive sequencing effort of all fractions throughout the CsCl gradient using quantitative SIP metagenomics [These results from two soils indicate that virus infection of AOA during nitrification is likely a common and dynamic process, with the potential to influence nitrification rates in soil. Although viruses were only identified from contigs representing partial viral genomes, analysis of individual marker genes and genome-wide similarity comparisons revealed that those infecting AOA are highly divergent from known viruses. While one genomics or alterSupplementary Information"} +{"text": "Fungal blood cultures are usually ordered when sepsis secondary to disseminated fungal infection is suspected. We aimed to analyze whether positive fungal blood cultures had an added clinical impact over other conventional microbiological tests.We performed a retrospective study of all patients for whom fungal blood cultures were performed for any indication at our institution from June 2018 to March 2022. We reviewed informatics database and medical records to assess the microbiological and clinical impact of positive fungal blood cultures during the initial admission. Assessment of clinical impact was analyzed using a list of predefined positive, negative, and no impact scenarios (Table\u00a01) based by the treating team\u2019s decisions.Candida spp. , followed by Histoplasma spp. , Cryptococcus spp. , and unidentified mold (Table\u00a01). The median time to positivity was 106 hours (IQR 79-177). Only 21 (16%) fungal blood cultures resulted in a change in management. Positive fungal blood cultures for fungi led to a positive clinical impact in 17 (13%), negative clinical impact in 4 (3%), and no clinical impact in 109 (84%). Most fungal blood cultures confirmed other conventional microbiological diagnosis or result was not acted upon . Organisms where fungal blood culture had a positive impact included C. albicans (n=4), and C. parapsilosis (n=3), while Cladosporium spp. (n=2) was the most frequent fungi with a negative clinical impact. Isolated fungi that had no clinical impact because of other confirmatory tests included Candida spp. (n=57), Histoplasma spp. (n=13) and Cryptococcus neoformans (n=10).In total, 4447 fungal blood cultures were performed in 3648 admissions during our study period. The overall positivity rate of fungal blood cultures was 6.4% (n=284), of which only 130 (2.9%) were positive for fungi. The most common isolated fungi were Most of positive fungal blood cultures for fungi resulted in no immediate clinical impact due to other microbiological tests available sooner or isolates thought to be non-clinically significant. Further studies should evaluate the long-term impact of fungemia and identify stewardship interventions to optimize clinical utility of fungal blood cultures.All Authors: No reported disclosures."} +{"text": "The fabricated composite NFs have been tested as a photocatalytic material to degrade methylene blue (MB) as a model dye under visible light. The introduced composite NFs have shown good photocatalytic activity compared with pristine TiO2 NFs; 100% and 50% of dye were degraded in 120 min for composite NFs and pristine TiO2 NFs, respectively. Furthermore, composite NFs demonstrated good stability for four cycles. In addition, the fabricated Cu-TiO2 NFs have shown good photocatalytic activity for the production of H2 from sodium borohydride.Simple and inexpensive electrospinning and hydrothermal techniques were used to synthesize titania nanofibers (TiO"} +{"text": "TP53 was most commonly mutated (30%) followed by PIK3CA (15%), AKT1 (11%), and CDKN2AIP (11%). We also identified significant intratumoral genomic heterogeneity, consistent with a branched evolution model, through multi-region exome sequencing of three distinct tumor regions from selected primary splenic tumors. These data provide new perspectives on the genomic landscape of this veterinary cancer and suggest a cross-species value for using HSA in pet dogs as a naturally occurring model of intratumoral heterogeneity.Cancer genomic heterogeneity presents significant challenges for understanding oncogenic processes and for cancer\u2019s clinical management. Variation in driver mutation frequency between patients with the same tumor type as well as within an individual patients\u2019 cancer can shape the use of mutations as diagnostic, prognostic, and predictive biomarkers. We have characterized genomic heterogeneity between and within canine splenic hemangiosarcoma (HSA), a common naturally occurring cancer in pet dogs that is similar to human angiosarcoma (AS). HSA is a clinically, physiologically, and genomically complex canine cancer that may serve as a valuable model for understanding the origin and clinical impact of cancer heterogeneity. We conducted a prospective collection of 52 splenic masses from 43 dogs presenting for emergency care with hemoperitoneum secondary to a ruptured splenic mass. Multi-platform genomic analysis included matched tumor/normal targeted sequencing panel and exome sequencing. We found candidate somatic cancer driver mutations in 14/27 (52%) HSAs. Among recurrent candidate driver mutations, Such heterogeneity presents challenges for uniform diagnosis, prognosis, and treatment planning for many cancers . IntratuTP53, PIK3CA, PTEN, PIK3R1 [PDGFRA, VEGFA, KIT, KDR), and copy number deletions [MYO16-PTK2, GABRA3-FLT1, and AKT3-XPNPEP1) [Angiosarcoma (AS), an aggressive cancer arising from vascular endothelium in anatomic sites including skin and viscera, is an important setting for the study of ITH. AS is rare in humans, but far more common in pet dogs. Tens of thousands of canine diagnoses occur annually, comprising 45\u201351% of splenic cancers \u201313. Shar, PIK3R1 , 13\u201316, B, PTEN) , 15, 16 XPNPEP1) , many ofSamples and sequencing platforms are summarized in Buffy coat and plasma were collected and stored in -20\u00b0C until nucleic acid isolation. Genomic DNA from 200uL buffy coat was isolated with DNeasy Blood and Tissue kit (Qiagen) according to manufacturer\u2019s protocol. Tumor nucleic acid extractions were performed using the Allprep DNA/RNA/miRNA Universal kit (Qiagen) according to manufacturer\u2019s protocol. Briefly, tumor tissue in Allprotect was first rinsed with sterile 1X phosphate buffered saline and minced with sterile scalpel. Thirty mg of the minced tissue was then homogenized using the Bullet Blender Bead lysis kit (NextAdvance) and the resulting supernatant was further homogenized with QiaShredder (Qiagen). The flow through was used for nucleic acid extraction. Quality and quantity of blood and tumor genomic DNA was performed using the Qubit Fluorometer 2.0 (ThermoFisher Scientific) and TapeStation genomic DNA assay (Agilent Technologies). Genomic DNA was stored in -20\u00b0C until sequencing library construction. RNA was extracted and stored in -80\u00b0C for future use as per manufacturer\u2019s instructions.in silico PCR. Sequencing libraries were constructed using droplet-based PCR amplification following the manufacturer\u2019s protocols for the ThunderBolts Cancer Panel with specific modifications (RainDance Technologies) as previously described [Targeted amplicon sequencing was performed on genomic DNA from matched blood and DNA from individual patient AllProtect tumor sections with the highest tumor content based on evaluation of adjacent FFPE. A custom canine HSA cancer amplicon sequencing panel consisting of 330 amplicons targeting exons and hotspot regions in 68 genes, with amplicon sizes ranging from 91\u2013272 bp was developed . Primersescribed . Paired-escribed . Sequencescribed and qualescribed . Sequencescribed . Custom escribed were useGenomic DNA from blood and all 3 tumor sections (AllProtect samples) from 4 HSA patients underwent whole exome sequencing using the Agilent SureSelectXT CD Canine All Exon V2 (Agilent Technologies), a community-based design that contains 982,789 probes covering 19,459 genes . The BEDBased on the recognized clinical, histologic, and cellular heterogeneity of canine HSA we sought to characterize its degree of inter- and intra-tumoral heterogeneity through a prospective multicenter study in whichTP53 was the most frequently mutated gene, with 11 mutations occurring across 8 patients (30%). Of 11 TP53 mutations, 8 were missense with most occurring in human-equivalent pathogenic hotspots in the DNA-binding domain. Single cases of splice acceptor (c.530-2A>C), stop gain (R296*), and frameshift (T243fs) mutations were identified. PIK3CA mutations were identified in 4 patients (15%). PIK3CA H1047L was the only recurrent point mutation, identified in 2 cases. The third and fourth cases bore H1047R and G1049R mutations. Amino acid 1047 is the most frequently mutated PIK3CA hotspot in human cancers, previously shown to be mutated in HSA (30\u201346% of cases) [AKT1 was mutated in 3 cases (11%) with two potentially pathogenic missense mutations, G37D and R23W, and one frameshift, L52fs, with unknown significance. Additional likely pathogenic mutations in single patients included CDKN2B R105Q and NRAS Q61R. Variants of unknown significance (VUS) include CDKN2AIP Q67R, a gene deleted in HSA via aCGH studies [ERBB2 splice region variant, FLT3 N703S, JAK3 M724T, PTEN H39fs, and PTPRB L1284P and S1965P. Of the 7 panel-sequenced benign cases, only Patient 37 (benign hyperplasia) bore a somatic SNV in a cancer gene\u2014a VUS impacting PTPRB. Patient 43 bore a likely pathogenic missense mutation at R789C in GNAS. These driver mutations and their frequencies resemble those identified in HSA in other studies [To identify somatic single nucleotide variants (SNVs), we utilized a custom canine cancer next generation sequencing amplicon panel with regf cases) , 16, 36. studies , an ERBB studies , 16, 37.Recurrent, potentially pathogenic mutations were identified in 24 matched normal/tumor and 3 tumor-only FFPE- or AllProtect-preserved splenic hemangiosarcomas. As indicated by colored boxes, missense and truncating mutations identified by amplicon panel-based sequencing (CCAP) with allele frequencies \u2265 0.1 are shown. Allele frequencies are indicated in colored boxes. Tumor content percentage and tumor grade are shown where available. Patient IDs with asterisks indicate samples that were also exome-sequenced.NRAS and PIK3CA mutations such as those described in HSA have been associated with MEK and PIK3CA inhibitor responses, respectively, in human cancers and may also be associated with such responses in canine cancers. Such responses, however, may also be dependent on intratumoral heterogeneity and will certainly be dependent on the ability to detect subclonal or low allele-frequency mutations. Basket and umbrella clinical trials that test such hypotheses in canine HSA will hold value broadly for understanding heterogeneous human cancers and will be especially valuable in less common human cancers such as angiosarcoma.Intratumoral genomic heterogeneity presents challenges to cancer precision medicine . For example, variability in subclonal frequency of driver mutations may modulate treatment response. Additionally, such variability means that any given biopsy utilized for genomic analysis may not contain driver mutations with predictive associations even if these mutations are highly abundant in major lineages in the tumor. This study has confirmed the presence of genomic variants and intratumoral heterogeneity in canine HSA that can facilitate hypothesis testing and methods development to meet these needs. For example, In order to more deeply explore heterogeneity, we next directly measured ITH via whole exome sequencing (WES) of three geographically distinct splenic sections from 4 patients along with their matching constitutional DNA from peripheral blood, achieving average sequencing coverage of 214x for tumors and 211x for normals. Patients with known pathogenic driver mutations according to panel-based sequencing and those with high tumor content according to pathology for all 3 sections were selected . Based oNRAS Q61R, was detected in all regions, although the hotspot TP53 Y210C was only detected in a single section. In Patient 16, 18/28 (64%) SNVs were shared in regions 2 and 3, including a missense mutation of unknown pathogenicity TP53 Y152S . No mutations were shared by all regions or between regions 1 and 3 and PIK3CA H1047L. In Patient 11, 61/181 (34%) SNVs occurred in 2 or more regions and 24 (13%) were shared among all regions and AKT1 G37D in Patient 22 (detected in CCAP but not WES likely due to the increased sensitivity of CCAP sequencing). Overall, of 8 total known pathogenic or likely pathogenic cancer driver mutations detected in these patients, half were shared among all distinct tumor regions from the same dog whereas the other half were present in only a single section. This variability in somatic mutation frequency across different geographic regions from the same tumor supports that significant genomic heterogeneity exists in HSA. Alongside cellular/physiologic heterogeneity (i.e. tumor content variability), this genomic heterogeneity holds implications for understanding HSA development and diagnosis while positioning HSA as a model system to understand and manage ITH.Analysis of intersecting somatic coding mutations across 3 tumor regions from 4 patients by WES identified substantial intratumoral genomic heterogeneity . Between 1 and 3 . For Pat 1 and 3 . Only re regions . Two driIntratumoral heterogeneity was assessed via whole exome sequencing of 3 distinct geographic regions from 4 canine HSA patients selected based on high average tumor content across multiple samples as well as presence of one more known or likely driver mutations from CCAP-based sequencing of the highest tumor content section. Individual patient oncoprints show coding, nonsynonymous mutations within each tumor section. Shared genomic events align vertically between the various tumor regions within a given patient. Each section is also annotated with tumor content assessment based on pathologic review in parentheses. All known pathogenic or likely pathogenic driver mutations are annotated and indicated in red text. TMB = tumor mutation burden.Phenotypic and genomic heterogeneity are well-established and increasingly well-understood features of human solid tumors that hold relevance for understanding cancer\u2019s genesis, its development, and its varied clinical behaviors. They also have bearing on cancer diagnosis via traditional diagnostics, like histopathology, and novel diagnostics such as genomic tumor profiling. Here we describe genomic heterogeneity in a common naturally occurring canine cancer, hemangiosarcoma (HSA), that has long been recognized as a clinically, physiologically, and histologically complex cancer. Through multi-platform (exome and gene panel) genomic analysis of 28 HSAs focused on identification of somatic SNVs, we identified marked inter- and intra-tumoral heterogeneity in canine HSA. First, we identified candidate somatic cancer driver mutations in 50% of cases. Notably, the majority of the 258 somatic SNVs detected in this cohort occurred at low tumor AFs < 10% , though all but one of the tumors bearing a known somatic driver contained at least one somatic mutation with an AF \u2265 10% . This obTP53 was the most commonly mutated gene (30%) followed by PIK3CA (15%), AKT1 (11%), and CDKN2AIP (11%), though frequencies of some detected mutations differ. Cross-study variation in this landscape, such as the lower rate of PIK3CA mutations identified in our cohort relative to others (15% versus 30\u201346%), likely reflects natural variation by HSA anatomic site, clinical characteristics, and breed in addition to variation in sequencing platforms and analysis approach. It underscores the need for expanded genomic study in very large cohorts. It also remains possible that other drivers are present in this cohort in regions not included in the panel in addition to copy number variation, translocations, and fusions. Large, integrated genomic studies that incorporate multiple sequencing platforms and mutation identification types across a diversity of breeds and anatomic sites will ultimately be necessary for refining our view of HSA\u2019s genomic landscape.These data also contribute to our growing understanding of HSA\u2019s genomic underpinnings. The genomic landscape in this cohort was similar to what has been previously described. NRAS, TP53, and PIK3CA were detectable in all sections from the same tumor at AFs ranging from 10\u201384%, supporting that in some cases, truncal driver mutations arise in clonal fashion and are universally present in the tumor cells . Also notable is that the cases with lower regional concordance contained at least one section with low TMB and low bioinformatically inferred tumor content even though histological review of tumor content may have been high Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S1 FigRepresentative images of hematoxylin and eosin-stained canine hemangiosarcoma sections adjacent to tumors sequenced in this study. 100x magnification with 50 \u03bcm scale bars shown on the lower left. (A) Patient 3. This tumor has low cell density with large cavernous blood-filled sinusoidal structures lined by monotonous endothelioid cells and separated by thin trabeculae. (B) Patient 12. This tumor has variable density with solid areas of tumor cells and disorganized collapsed blood-filled clefts as well as areas with large cavernous blood-filled sinusoidal structures. The atypical endothelioid cells have plump nuclei with anisokaryosis.(TIF)Click here for additional data file.S2 Fig(TIF)Click here for additional data file.S3 FigFor each of the cases with three independently sequenced tumor sections, the Pearson correlation coefficient was calculated for: A) Median variant allele fraction (AF) versus histologically and bioinformatically determined tumor content estimates; and B) Tumor mutation burden (TMB) versus histologically and bioinformatically determined tumor content estimates.(TIF)Click here for additional data file."} +{"text": "Perturbation-based training (PBT) is a balance training method presenting a high-challenge to balance which is extremely effective when compared to conventional training approaches. Common PBT methods use rapid treadmill belt translations with varying numbers of perturbations (20-1400 perturbations over 1-8wks). Importantly, the joint loads experienced during a stumble may be high enough to feasibly fracture bone (up to 12.7 Body Weights). However, the contact loads experienced during PBT are unknown. Because of increasing the prevalence of PBT it is necessary to specifically evaluate the range of joint loads experienced during PBT. Twelve participants completed a single PBT session of 24 perturbations. During both training and testing, participants movements were measured using a motion capture system which tracks body movements and records the forces under the feet. Hip joint contact loads were determined using Computational Musculoskeletal Modelling utilizing open-source software OpenSim. These techniques estimate the magnitude and pattern of force development of individual muscles and subsequently estimate the internal loads experienced by the hip joint.Hip joint contact loads were 4.90 \u00b1 1.27 BW which is substantially lower that those previously reported by Graham et al. (2016) and Bergman et al. (2004) and is lower than the 5.5BW spontaneous fracture load boundary estimated by Schileo et al. (2014). Comparing the initial perturbation to the final perturbation revealed a 22% reduction in contact loads. We conclude that PBT performed using rapid translations on a treadmill are likely safe but suggest caution for individuals with poor bone mineral density or reduced neuromuscular function."} +{"text": "For the genotype\u2019s average mean, chlorophyll content decreased by 10.5% under drought at WDSF. Net photosynthetic rate (Pn), stomatal conductance (gs), and transpiration rate (E) were reduced at WDA by 53%, 80.8%, and 61.4% and at WDSF by 43.75%, 57.7%, and 36%, respectively, while relative water content (RWC) reduced by 16.48%, on average, for both stages. G10 and G6 showed adaptability when water scarcity occurred at an early (WDA) or later stage (WDSF), respectively, providing insights into using germplasm resources to cope with the drought effect.Drought affects common bean productivity, and the severity of its impact is expected to increase due to climate change. The use of versatile genotypes could contribute to securing future bean production. This study investigates the adaptability of 10 common bean genotypes of indeterminate growth type under water scarcity conditions by measuring agronomic and physiological parameters. The evaluation occurs under irrigation treatments applied at two different phenological stages (anthesis (WDA) and seed filling initiation (WDSF)). The recorded adaptabilities of the genotypes (G) showed that G10 produced the highest overall seed yield in the normal irrigation (NI) (197.22 g plant Phaseolus vulgaris L.) is one of the most important pulses worldwide due to its high protein content, fiber, and other essential minerals for humans \u00d7 100.Data were subjected to over-year analysis of variance (ANOVA) as a quadruplicated split-plot design with irrigation levels (I) as the main plot factor and genotypes (G) as sub-plot. The Shapiro\u2013Wilk test for normality showed that the variables were normally distributed, whereas the ANOVA\u2019s assumptions for the equality of the error variances and residual normality were met by Levene\u2019s test and the normal quantile\u2013quantile (QQ) plot method, respectively . The difGenotype plus genotype \u00d7 environment (GGE) biplot analysis was used for analyzing G \u00d7 Y interactions and ranking cultivars for yield and stability . The advThe evaluation revealed two genotypes with high versatility under drought conditions that could contribute to common bean adaptability in the upcoming climate change scenario and could be used directly in cultivation or as starting material in a breeding project. Specifically, G10 showed good productivity when water deficit occurs at an early stage of reproductive development , while G6 showed adaptability when drought events happen at a later stage . This is probably connected with the ability of each genotype to maintain sink strength and the balance of source availability under water-limiting conditions. Further study of genotype \u00d7 environment interactions is important for revealing genotypes showing good performance and stability across conditions with adverse conditions and provide useful information for breeders and agronomists."} +{"text": "Sleep disturbances in people living with cognitive impairment (CI) may impose a great burden on caregivers. We examined the relationship between objective and subjective sleep measures in people living with CI and caregiver depression and mastery via a secondary analysis of The Healthy Patterns Clinical Trial (NCT03682185) baseline data (n=170). Objective sleep variables included total sleep time and sleep efficiency derived from 3 nights of actigraphy. Subjective sleep measures included PROMIS Sleep Related Impairment, Pittsburgh Sleep Quality Index, and Epworth Sleepiness Scale. Caregiver measures included Center for Epidemiological Studies Depression Scale and Caregiver Mastery Scale. People living with CI were female (67%), Black (80%), with mean age 73.4 \u00b1 8.7. Caregivers were female (81%), family caregivers (80%) with mean age 56.5 \u00b1 14.7. We used multiple regression analysis, adjusting for cognition, and examined if there were differences by caregiver gender. Poorer subjective sleep quality was significantly associated with more caregiver depression . There were no significant sleep predictors for caregiver mastery; however, there was a moderating effect of gender on the association between subjective sleep quality and caregiver mastery. Female caregivers had increased caregiver mastery compared to males when the person living with CI had better sleep quality . This study found that people living with CI sleep characteristics differentially influence caregiver outcomes. Sleep should be assessed using a combination of objective and subjective sleep measures in people living with CI to inform providers when interventions are needed."} +{"text": "Cardiac electrophysiology is a complex system established by a plethora of inward and outward ion currents in cardiomyocytes generating and conducting electrical signals in the heart. However, not only cardiomyocytes but also other cell types can modulate the heart rhythm. Recently, cardiac macrophages were demonstrated as important players in both electrophysiology and arrhythmogenesis. Cardiac macrophages are a heterogeneous group of immune cells including resident macrophages derived from embryonic and fetal precursors and recruited macrophages derived from circulating monocytes from the bone marrow. Recent studies suggest antiarrhythmic as well as proarrhythmic effects of cardiac macrophages. The proposed mechanisms of how cardiac macrophages affect electrophysiology vary and include both direct and indirect interactions with other cardiac cells. In this review, we provide an overview of the different subsets of macrophages in the heart and their possible interactions with cardiomyocytes under both physiologic conditions and heart disease. Furthermore, we elucidate similarities and differences between human, murine and porcine cardiac macrophages, thus providing detailed information for researchers investigating cardiac macrophages in important animal species for electrophysiologic research. Finally, we discuss the pros and cons of mice and pigs to investigate the role of cardiac macrophages in arrhythmogenesis from a translational perspective. Cardiac arrhythmias affect millions of individuals worldwide and are a major cause of morbidity and mortality, thereby causing a significant medical but also socioeconomic burden . The pat+CX3CR1+F4/80high and heart valves 7.25) and the fetal liver (at E8.25) migrate into the heart where they persist as \u201cresident cardiac macrophages\u201d by feration . In the feration . Moreovet valves .\u2212MHC-IIlow) or partly (CCR2\u2212MHC-IIhigh) replenished through in situ proliferation with negligible monocyte input -4 triggered inflammatory response (high (CCR2+CX3CR1lowGr1+) and noninflammatory Ly6Clow (CCR2\u2212CX3CR1highGr1-) monocytes mouse model are preferentially recruited to sites of injury, where they enhance inflammation by promoting further monocyte and neutrophil recruitment and cause cardiomyocyte hypertrophy, interstitial fibrosis and adverse cardiac remodeling resulting in poor outcome, e.g. after myocardial infarction (\u2212MHC-IIlowLy6Clow and CCR2\u2212MHC-IIhighLy6Clow) demonstrate robust proangiogenic properties, suppress inflammation by inhibiting monocyte recruitment and enhance cardiac repair , thereby causing collagen deposition and diastolic dysfunction activity and IL-1\u03b2. Intriguingly, overexpression of Osm was also observed in cardiac patient hearts suggesting a role in heart disease (+ (FOLR2+ and/or TIMD4+ and/or LYVE1+) macrophages originate from both yolk sac and fetal monocyte precursors, and are the most transcriptionally conserved subset (+TIMD4\u2212 macrophages appear to be related to resident macrophages and are associated with cardiovascular remodeling. LYVE1+FOLR2+ macrophages are monocyte-derived and express the chemoattractant cytokine genes CCL13 and CCL18. Patients with dilated cardiomyopathy showed reduced numbers of LYVE1+ resident macrophages (subsets expressed FOLR2 and HSPH1) and an increased number of inflammatory macrophages (\u2212FOLR2\u2212MERTK- macrophages are associated with antigen presentation (Single-cell RNA sequencing (scRNA-seq) allows precise identification of macrophage subpopulations in the heart. In heart failure mice, two CCR2 disease . scRNA-sd subset . In the nd KLF2) . In contentation .in vitro . In addition, T helper cell (Th2)-produced cytokines such as IL-4 and IL-13, activate M2 macrophages, which control the proliferation of T cells and attenuate the immune response demonstrating that pro- and anti-inflammatory processes of cardiac macrophages happen simultaneously and disrupt homeostasis . Immediately following the AP upstroke, a transient repolarization produced by an outward potassium current (Ito) follows (phase 1). Phase 2 represents a slowly decaying plateau, resulting from an equilibrium of the delayed-rectifier potassium current with slow, rapid, and ultrarapid activation kinetics , the Na+-K+-ATPase current (INKA), the late sodium current and a simultaneous prominent calcium influx . Phase 3 is characterized by a rapid repolarization, resulting from the inactivation of calcium channels and the activation of an inward rectifier potassium current (IK1). The resting state (phase 4), which is also mainly driven by the potassium current IK1 establishes the resting membrane potential (RMP). Under physiologic conditions, the APD determines the effective refractory period (ERP), which is defined as the shortest time interval needed before a new stimulus can depolarize the cell again causing another AP in individual cardiomyocytes Nerbonn. The durother AP .If) is responsible for the membrane hyperpolarization , play a fundamental role in both depolarization of SAN and AVN cells and in counteracting repolarization despite lacking a clear plateau phase are most abundantly expressed in SAN and AVN where this current contributes to the diastolic depolarization and in the atria where it hyperpolarizes the cell contributing to the phase 3 repolarization and reentry (acting as substrate) which are reviewed in detail elsewhere . Here, w2+ channels ) or phase 3 (in the context of Ca2+ overload and activated Na+/Ca2+ exchange current (NCX) resulting in Na+ influx) . DADs usmulation .Aside from altered impulse formation, abnormal impulse conduction represents another main mechanism of arrhythmogenesis . This caThe major hallmark of structural remodeling and subsequently maintenance of arrhythmia is cardiac fibrosis, characterized by an imbalance between the generation and degradation of extracellular matrix (ECM) . Under mThe AP propagation depends on gap junctions, transmembrane proteins that mediate cell-to-cell coupling . Gap junCardiac electrophysiology is modulated by the autonomic nervous system (ANS) consisting of sympathetic and parasympathetic nerves . An imbaIt has been suggested that macrophages play an important role in arrhythmogenesis since they can produce a number of cytokines which have been linked to cardiac remodeling. However, only recently a few studies have been published that clearly demonstrate specific macrophage-dependent mechanisms regulating physiologic conduction as well via gap junction channels (established by connexins) which is an essential prerequisite for electrical impulse propagation. Modifications of connexins such as phosphorylation or altered distribution of gap junctions result in reduced conduction velocity or altered pathways of conduction and anisotropy leading to a proarrhythmic substrate predisposing to reentry and human (CD68+CD163+) and that they are functionally coupled to cardiomyocytes via Cx43. They revealed that macrophages coupled to cardiomyocytes show spontaneous rhythmic depolarizations whereas cardiomyocytes coupled to AV nodal macrophages show an elevated resting membrane potential and a shortened action potential, which facilitates electrical conduction in the AV node. These findings were further supported by several in vivo models: 1) using an optogenetics approach they were able to specifically depolarize AV nodal macrophages in situ which resulted in enhanced electrical conduction through the AV node illustrated by a higher number of conducted stimuli at the Wenckebach point. 2) In a transgenic mouse model with macrophage-specific knockdown of Cx43 (Cx3cr1wt/CreER Cx43\u00a0fl/fl) they demonstrated an increased Wenckebach cycle length and AV node refractory period indicating that disruption of the macrophage-cardiomyocyte interaction results in impaired electrical conduction in the AV node. These findings were further confirmed by 3) a transgenic mouse model that congenitally lacks macrophages (Csf1op). 4) Inducible macrophage depletion in CD11bDTR mice finally showed progressive AV block within 1\u20132\u00a0days. All these findings illustrate that cardiac resident macrophages facilitate electrical conduction in the AV node by electrical coupling to cardiomyocytes via Cx43 stress (pulmonary artery banding leading to RV pressure overload), cardiac macrophages protect the heart from arrhythmia and sudden cardiac death (SCD) by maintaining proper electrical conduction through gap junctions . Depleti3cr1eYFP/+) macrophages and reported a membrane resistance of 2.2 \u00b1 0.1 G\u03a9, a capacitance of 18.3 \u00b1 0.1 pF, and a resting membrane potential of -39.6 \u00b1 0.3\u00a0mV an elevated number of recruited macrophages have been detected in the infarct border zone where they form gap junctions with adjacent cardiomyocytes, a finding that was confirmed in a mouse model of MI . Recruitl KCa3.1 . This fanduction have been described in left and right atria with several remodeling mechanisms being regulated by macrophages . In patiht atria .ICa,L in atrial myocytes leading to AF (Further mechanistic studies in lipopolysaccharide (LPS)-treated mouse and canine models revealed that recruited macrophages induce electrical remodeling by secreting pro-inflammatory cytokines, including TNF-\u03b1, IL-1\u03b2, and IL-6 Figure . In thesng to AF .+ macrophages and increased expression of IL-1\u03b2 and TNF-\u03b1 in LA indicating proarrhythmic structural and electrical remodeling together with an inflammatory response , reduced expression of Cx43 as well as enlarged atria, enhanced atrial fibrosis and increased levels of reactive oxygen species, ultimately leading to an increased susceptibility for AF revealed that macrophages play a pivotal role in cardiac remodeling as they may have both profibrotic and antifibrotic functions . CD68+ oe for AF . As fibre for AF .high macrophages release anti-fibrotic cytokines like Osm to inhibit the conversion of fibroblasts to myofibroblasts. Ly6Clow macrophages in contrast produce profibrotic cytokines such as TGF-\u03b21 and IL-10 to promote fibrosis and indirect effects on cardiomyocytes were detected in right atrial appendages which was associated with increased atrial gene expression of procollagen and B-type natriuretic peptide (BNP) and atrial fibrosis it is another hint towards a direct role of recruited cardiac macrophages in proarrhythmic structural remodeling and reparative (expressing osteopontin) macrophages. Although this work illustrates the association of macrophages with fibrotic remodeling, potential conclusions on arrhythmogenesis must be drawn with caution as 1) there was no direct evidence for arrhythmias in these mice, 2) direct macrophage function/effects were not investigated, and 3) a direct causal role for macrophages remains elusive.Arrhythmogenic cardiomyopathy is an inherited disease characterized by progressive structural remodeling ultimately leading to arrhythmias and SCD . In mice\u2212) macrophages and recruited monocyte-derived (CCR2+Ly6Chigh) macrophages may have distinct and at least partly antagonistic effects it has been indicated that cardiac resident or catecholaminergic polymorphic VT (CPVT) where the autonomic innervation has been identified as key factor in arrhythmogenesis can be further grouped into two numerically dominant Ly6ClowGr1\u2212MHC-IIhigh and Ly6ClowGr1\u2212MHC-IIlow subsets, and the Ly6ChighGr1+ subset. Furthermore, resident macrophages can be identified as F4/80high, whereas recruited macrophages (CCR2+CX3CR1\u2212) are F4/80lowCD11bhigh. Recruited macrophages with a proinflammatory phenotype also express CD11c, whereas those with a reparative phenotype can be identified as CD163+CD206+ (As described in pression . Residen3+CD206+ . Other c3+CD206+ .high) and reparative (CD206+) phenotypes using human recombinant GM-CSF and M-CSF, respectively , the ICau phase , and IK in mice .Ito) current in pigs and from Ito current in humans (Pigs have been increasingly used in cardiovascular research as they closely resemble the human cardiac anatomy and (electro)physiology . FurtherIto) . Major dn humans .Although more research on specific porcine macrophage subsets is warranted to allow a more precise classification, pigs have several obvious advantages over other species especially regarding the resemblance to human electrophysiology and immunology, which make them an ideal choice as large animal species for studying cardiac macrophages in arrhythmogenesis.Inflammation is thought to crucially contribute to the development of an arrhythmogenic substrate. Emerging data underlines vital roles of distinct cardiac macrophage subsets for regulating proarrhythmic electrical, structural, or autonomic remodeling. Given their remarkable plasticity, multiple origins and phenotypes, further analysis of the functionality of macrophage subsets in arrhythmogenesis is clearly necessary. A better understanding of arrhythmia mechanisms will reveal new potential therapeutic targets allowing the development of innovative therapeutic strategies for patients suffering from arrhythmias. To achieve this goal, initial findings on macrophage-mediated arrhythmia mechanisms obtained in rodent models need to be validated in preclinical close-to-human large animal models prior to clinical application. As they share close similarities with humans regarding cardiovascular anatomy, electrophysiology and immunology we propose pigs as suitable large animal species for translational research on cardiac macrophages and their role in arrhythmias."} +{"text": "After reporting a single wild poliovirus (WPV) type 1 (WPV1) case in 2021, Pakistan reported 14 cases during April 1\u2013July 31, 2022. Pakistan and Afghanistan are the only countries where endemic WPV transmission has never been interrupted immunization. The World Health Organization (WHO) and UNICEF estimated Pakistan\u2019s 2021 national polio vaccination coverage at 83% and 2 SNIDs (in January and June) targeting children aged <5 years have been conducted to date in 2022 using bOPV. SNIDs took place in designated, high-risk districts for poliovirus transmission and other priority areas for the polio program mostly in the provinces of Khyber Pakhtunkhwa, Balochistan, Sindh, and Punjab. Limited SIAs were conducted in response to identification of WPV1 cases and environmental isolates in March, April, and June. An NID was conducted in August; another is planned for November, and an SNID is planned for October 2022.During SIAs conducted in 25 very high-risk districts,Acute flaccid paralysis surveillance. Detection of two or more cases of nonpolio acute flaccid paralysis (AFP) per 100,000 children and adolescents aged <15 years per year is an indicator of adequately sensitive polio surveillance.Environmental surveillance. Laboratory testing of sewage samples routinely collected at designated sites supplements AFP surveillance in facilitating timely detection of circulating polioviruses. Pakistan has 77 environmental surveillance sampling sites. During 2021, 65 (8%) of 833 sewage samples tested positive for WPV1 compared with 407 (52%) of 786 samples tested in 2020. In 2022, to date, 13 (2%) of 748 samples have tested positive for WPV1, including eight from Khyber Pakhtunkhwa province, four from Punjab province, and one from Islamabad. The earliest isolates detected in samples collected from environmental surveillance sites in Bannu district (Khyber Pakhtunkwa) in April 2022 were orphan viruses , indicating gaps in AFP surveillance sensitivity; subsequent isolates were genetically linked to WPV1 cases detected in North Waziristan.Epidemiology of poliomyelitis cases. During 2021, a single Pakistan WPV1 case was reported in Killa Abdullah, Balochistan, compared with 84 cases reported from several provinces in 2020 and 147 cases reported during 2019 ; 87% had never received OPV through essential immunization (zero-dose children), and 13% had received 1\u20133 OPV doses through essential immunization. Genetic analysis of the viruses identified in the WPV1 cases indicated that all belong to a single genetic cluster (groups of polioviruses sharing \u226595% sequence identity in the region coding the VP1 capsid protein). However, three additional genetic clusters were identified from environmental surveillance isolates during January 2021\u2013July 2022, again an indication of AFP surveillance gaps, although only one genetic cluster has been detected since June 2021.Transmission of cVDPV2 from several emergences in Pakistan resulted in 165 cVDPV2 cases during July 2019\u2013April 2021 . In the most recent case, the patient had paralysis onset on April 23, 2021 Figure Figure .The number of WPV1 cases and areas of poliovirus transmission identified in Pakistan declined markedly during January 2021\u2013July 2022 compared with the preceding 2 years. The limited genetic divergence among WPV1 isolations since 2020 suggests that the reduction in cases and apparent geographic scope of virus spread are likely reflective of a decrease in WPV1 circulation during the reporting period. Disruptions to implementation of polio eradication activities because of the COVID-19 pandemic have been ameliorated since late 2020 where wild poliovirus type 1 (WPV1) transmission has never been interrupted.WPV1 cases in Pakistan decreased from 147 in 2019 and 84 in 2020 to a single case in 2021 but increased to 14 cases in 2022 as of July 31. These 14 WPV1 cases are clustered among children in southern Khyber Pakhtunkhwa province, many of whom have never received poliovirus vaccine (zero-dose children).Ensuring the highest quality vaccination activities in priority areas of Pakistan will enable the polio program to improve the chances of interrupting ongoing transmission of WPV1."} +{"text": "Anorexia nervosa (AN) represents a severe mental disorder associated with cardiovascular complications leading to morbidity and mortality. Abnormal functioning of autonomic nervous system, particularly sympathetic nervous system, plays a crucial role in AN-linked psychopathology and cardiovascular diseases; however, the pathomechanisms are still unclear.Thus, we studied sympathetic arousal in response to mental stress using conventional parameters, and for the first time by spectral analysis of electrodermal activity with aim to detect non-invasive biomarkers for cardiovascular risk assessment already in adolescent AN patients.Twenty-five AN girls were examined (14.8\u00b10.4 yr.) and age/gender matched controls (15.1\u00b10.3 years). Electrodermal activity (EDA) was continuously recorded at rest (5 min.) and in response to Go/NoGo test (5 min.). Evaluated parameters: skin conductance level (SCL) and spectral parameter of EDA in the sympathetic frequency band (EDASymp). EDA reactivity was calculated as percentual change (%) of SCL and EDASymp in response to stressor.The AN group had significantly reduced SCL and EDASymp compared to controls during baseline and in response to Go/NoGo test . The EDASymp index reactivity was significantly lower in AN group compared to control (p=0.034).Our study revealed resting sympathetic underactivity associated with lower reactivity to mental stressor indexed by EDA parameters in adolescent AN patients. This altered pattern of sympathetic arousal could play important role as a pathomechanism leading to cardiovascular complications in AN. It seems that EDA indices represent potential non-invasive biomarkers to detect AN-linked cardiovascular risk already at adolescent age.This study was funded by the Slovak Scientific Grant Agency under grants VEGA 1/0044/18 and VEGA 1/0190/20 and Ministry of Health of the Slovak Republic under the project registration number 2018/20-UKMT-16."} +{"text": "A 42 year old obese man with severe aortic regurgitation associated with bicuspid aortic valve and left ventricular dilatation presented with a hypertensive crisis (blood pressure [BP] 228/117\u00a0mmHg) and chest pain. Physical examination showed an ankle brachial index (ABI) of 0.6 and absent femoral pulses in both lower limbs. Computed tomography angiography showed an ascending aortic diameter of 4.7 cm and aortic coarctation with a critically stenosed aorta and prominent internal mammary arteries providing the collateral circulation. With extracorporeal circulation, reconstruction of the bicuspid aortic valve and the aortic root was done by means of the hemi-Yacoub technique and extra-aortic annuloplasty using a Dacron ring, as well as plication of the fused cusp. Moreover, an ascending to descending aortic bypass with a 19 mm expanded polytetrafluoroethylene graft was inserted after cleavage of the dorsal pericardium. The post-operative course was uneventful. The patient recovered distal pulses and had a normal ABI of 1.0 bilaterally. The patient had improvement in his hypertension (BP 133/81\u00a0mmHg) and reduced requirements for antihypertensive medications."} +{"text": "This nationwide population-based study analyzed the outcomes of local treatment (i.e. stereotactic body radiotherapy [SBRT] or metastasectomy) or systemic therapy for oligometastatic disease (OMD) in patients with esophagogastric cancer in The Netherlands.Between 2015 and 2016, all patients in The Netherlands with esophagogastric cancer and synchronous or metachronous OMD were eligible for inclusion. Patients who underwent local treatment of OMD (SBRT or metastasectomy) and/or systemic therapy were included. OMD was defined as distant metastases in 1 organ or 1 extra-regional lymph node region. The primary outcomes were overall survival (OS) and independent prognostic factors for OS. OS was calculated from diagnosis of OMD. Prognostic factors for OS were analyzed using a multivariable Cox proportional hazard model.A total of 594 patients were included, of whom 83 underwent local treatment for OMD alone, 22 local treatment plus systemic therapy, and 489 systemic therapy alone. Median OS after local treatment for OMD alone was 16.0 months, local treatment plus systemic therapy 22.7 months, and after systemic therapy alone 8.5 months. Improved OS was independently associated with local treatment for OMD alone or combined with systemic therapy as compared with systemic therapy alone and a controlled primary tumor. Worse OS was independently associated with worse performance scores , poorly or undiffertumor as compared with good or moderadifferentiated tumor , and peritoneal as compared with lymph mode metastases .Local treatment of OMD alone or combined with systemic therapy was independently associated with improved OS as compared with systemic therapy alone in this population-based cohort study in The Netherlands. Randomized controlled trials are warranted to confirm these results. Gastric and esophageal cancer are the 5th and 7th most common cancers worldwide and the incidence of esophageal cancer is rapidly rising In a small portion of metastatic patients, distant metastases are limited in number and distribution, so-called oligometastatic disease (OMD) However, the applicability and generalizability of the currently available data from the literature is unclear since clinical trial results cannot always be reproduced in the real-world setting due to strict selection criteria This study included patients registered in the Netherlands Cancer Registry (NCR). The NCR is the only national oncological registry in The Netherlands and provides cancer statistics among all 17.4 million residents. According to the Central Committee on Research involving Human Subjects, this study did not need approval by an institutional review board in The Netherlands. The study was approved by the Privacy Review Board of the Netherlands Cancer Registry and the scientific committee of the Dutch Upper GI Cancer Group (DUCG). The study was reported according to the guidelines of The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Statement [22].Consecutive patients with synchronous or metachronous metastatic esophagogastric cancer were identified from the NCR between 2015 and 2016 without evidence of locoregional recurrence at the time of OMD detection. Treatment of OMD was categorized into 1) local treatment alone (i.e. SBRT and/or metastasectomy); 2) local treatment plus systemic therapy (i.e. chemotherapy or targeted therapy); 3) systemic therapy alone. The administration of systemic therapy was divided into before or after local treatment of OMD. The first-line systemic therapy regimen administrated after the diagnosis of current OMD was analyzed .The primary outcomes of this study were OS and prognostic factors for OS. OS was defined as the time interval between the diagnosis of OMD and death or end of follow-up. Vital status was obtained through annual linkage with the municipal population registers and was last updated on January 31, 2021. Prognostic factors for OS were expressed using hazard ratios (HRs) with 95% confidence intervals (CIs). Kaplan-Meier curves were constructed for OS and independent prognostic factors for OS and were compared using log-rank test.T test. Non-parametric data were presented as median with interquartile range (IQR) and compared using Mann Whitney U test. Categorical data were presented as frequencies with proportions and compared using Fisher\u2019s exact test. Factors previously identified in literature Parametric data were presented as mean with standard deviation (SD) and were compared using Student\u2019s Between 2015 and 2016, 4265 patients with synchronous or metachronous metastatic esophagogastric cancer were identified from the NCR, of whom 594 patients who underwent local treatment or systemic therapy for OMD were included. First, the 105 patients undergoing local treatment for OMD with or without systemic therapy will be described. Subsequently, the 489 patients undergoing systemic therapy alone for OMD .Fig. 1FlThe 105 included patients were generally male (71%) with a mean age of 64\u00a0years (SD: \u00b18) and mostly had a WHO performance score of 0\u20131 at the time of treatment (62%). The primary tumors were predominantly adenocarcinomas (80%) located in the distal third of the esophagus (57%). The predominant clinical tumor stage was cT3 (66%) and nodal stage cN1 (45%). For patients who underwent primary tumor resection (n\u00a0=\u00a074), the predominant pathological tumor stage was pT3 (45%) and nodal stage pN0 (45%).Most patients had metachronous OMD . OMD was located in 1 distant organ (79%), 1 extra-regional lymph node region (12%), or the peritoneum (9%). The median disease-free interval for metachronous OMD was 17\u00a0months (IQR: 14\u201324) after diagnosis of the primary tumor. OMD was confirmed with pathological assessment (71%) or repeated follow-up imaging 29\u00a0%, .Table 1PPrimary tumor treatment consisted of surgery in 74 patients (71%), definitive chemoradiotherapy in 12 patients (12\u00a0%), or no primary tumor treatment in 19 patients (17%). Treatment of OMD consisted of local treatment alone in 83 patients (79%), including SBRT alone in 34 patients (33%), metastasectomy alone in 35 patients (32%), or both metastasectomy and SBRT in 14 patients (14%). Local treatment of OMD was combined with systemic therapy in 22 patients (21%), including metastasectomy plus systemic therapy in 14 patients (14%), SBRT plus systemic therapy in 7 patients (7%), or both metastasectomy and SBRT plus systemic therapy in 1 patient (1%). Systemic therapy was predominantly administrated before local treatment of OMD (73%) and generally consisted of 2 chemotherapy agents (68\u00a0%). The most common chemotherapy regimen consisted of capecitabine and oxaliplatin 36%, .Table 2TA total of 64 patients underwent metastasectomy. Metastasectomy was more commonly applied than SBRT for OMD in the liver (80%), the extra-regional lymph nodes (67%), or the peritoneum (100%). A total of 56 patients underwent SBRT. Applied SBRT schedules are provided in Patients with synchronous as compared with metachronous OMD less often underwent primary tumor resection (47% versus 87%), more often underwent local treatment of OMD plus systemic therapy (37% versus 10%), and had extra-regional lymph node oligometastases (19% versus 2%). Patients with metachronous as compared with synchronous OMD more often underwent local treatment of OMD alone (90% versus 63%) and had brain oligometastases , synchronous OMD , liver metastases , and an uncontrolled primary tumor as compared with patients who underwent local treatment for OMD with or without systemic therapy .The median follow-up time for patients undergoing local treatment for OMD with or without systemic therapy was 49.8 months (IQR: 37.2-55.0) and for patients undergoing systemic therapy alone was 59.0 months (IQR: 50.0-62.0). The median OS after local treatment of OMD plus systemic therapy was 22.7 months (95% CI: 14.7-42.6), versus 16.0 months (95% CI: 12.7-21.8) after local treatment of OMD alone, and 8.5 months (95% CI: 7.9-9.6) after systemic therapy alone .Fig. 2OvIn multivariable analysis , worse OImproved OS was independently associated with local treatment of OMD alone or combined with systemic therapy as compared with systemic therapy alone , and a controlled primary tumor versus uncontrolled primary tumor . However, these results must be interpreted with care because selection may have resulted in a potential overestimation of OS after local treatment of OMD because patients with favorable patient- and tumor characteristics were more often selected for treatment (i.e. confounding by indication) The benefit of local treatment of OMD plus systemic therapy over systemic therapy alone has been previously suggested by a phase II non-randomized trial by Al-Batran et al. [15]Although several non-randomized studies have suggested excellent OS in patients undergoing local treatment of OMD plus systemic therapy In addition to the German RENAISSANCE trial, , several phase 3 trials are currently investigating the benefit of local treatment for OMD plus systemic therapy over systemic therapy alone However, none of these studies have incorporated immunotherapy in the treatment algorithm for OMD, although several studies have shown improved survival outcomes for patients with esophagogastric cancer treated with immunotherapy in the first-line palliative setting Certain limitations apply to this study that warrants caution for the interpretation of results. First, no additional prognostic factors could be analyzed in the multivariable Cox proportional hazard model because of the risk of overfitting given the relatively limited sample size https://www.OMECproject.eu). OMEC is a consortium of 50 esophagogastric cancer expert centers across 16 countries in Europe. Studies of the OMEC-project include a systematic review of definitions of esophagogastric OMD (OMEC-1 The OligoMetastatic Esophagogastric Cancer (OMEC) project aims to achieve consensus on the definition and treatment of oligometastatic esophagogastric cancer or a combined approach consisting of radial local treatment of OMD plus systemic therapy (e.g. chemotherapy) . However, our results are most likely biased. Therefore, randomized controlled trials are warranted to confirm these results.The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Dr. van Laarhoven reports consultant or advisory role: BMS, Dragonfly, Lilly, Merck, Nordic Pharma, Servier; research funding and/or medication supply: Bayer, BMS, Celgene, Janssen, Incyte, Lilly, Merck, Nordic Pharma, Philips, Roche, Servier; Dr. Verhoeven reports grants from Bristol-Myer Squibb and Roche, outside the submitted work; Dr. Haj Mohammad reports personal fees from BMS, Lilly, MSD, Servier, and Astra Zeneca, outside the submitted work; the other authors have nothing to disclose.."} +{"text": "It has been well established that depressive disorders including perinatal depression are very heterogeneous, which partly explain the ineffectiveness of available treatments for many patients. Recent innovations in data science can help elucidate the nature of perinatal depression especially the heterogeneity in its presentation.The present study aime to elucidate heterogeneous subtypes of PND and assess the effectiveness of a multicomponent cognitive behavioral therapy (CBT) across heterogenous subtypes of PND.This study was conducted in 2005 in two rural areas of Rawalpindi, Pakistan. Out of a total of 3,898 women, 903 pregnant women were identifed with PND (using DSM-IV) and randomly assigned to intervention and control group. Baseline assessments included interviewer admininstered Hamilton Depression Scale (HDS) and social risk factors. Follow-up assessments were conducted at 6 months and 12 months post-intervention. Principle component analysis was run to reduce dimensionality of the HDS. Two step cluster analysis was then run to elucidate subtypes of PND using the dimensional scores. Thereafter, effectiveness of CBT was compared across these subtypes of PND using multilevel modelling.Principle component analysis revealed a four component solution for the Hamilton depression rating scale. Using these dimensional scores, cluster analysis (average silhouette= 0.5) revealed a parsimonius four cluster soultion of participants with mild PND symptoms (n=326); predominant sleep problems (n=311) c) predominant atypical symptoms (n=80) and d) comorbid depressive and anxiety symptoms (n=186). CBT yielded moderate effect sizes across all these subtypes of PND (cohen\u2019s d > 0.8).Multicomponent CBT is effective across hetergeneous presentations of PND.No significant relationships."} +{"text": "The Advisory Committee on Immunization Practices (ACIP) recommends that all persons aged \u22655 years receive 1 booster dose of a COVID-19 vaccine after completion of their primary series.https://vsafe.cdc.gov/en/). Health surveys sent daily during the first week after administration of each dose include questions about local injection site and systemic reactions and health impacts.The v-safe platform allows existing registrants to report receipt of a COVID-19 booster dose and new registrants to enter information about all doses received and completed at least one daily health survey after receiving their second booster dose and at least one survey after a previous vaccine dose. Among registrants who received homologous mRNA vaccination, the odds of reporting an adverse reaction or health impact after receiving the second booster dose versus previous doses were compared using a multivariable generalized estimating equations model that accounted for demographic variables and repeated measures. Comparisons of adverse reactions and health impacts by vaccine dose were restricted to persons who received homologous mRNA vaccination because previous studies observed different patterns of reporting among recipients of heterologous mRNA and among recipients of homologous Ad26.COV2 (Johnson & Johnson [Janssen]) booster vaccination . The median registrant age was 67 years; 173,525 (60.6%) were female. In the week after receipt of the second booster dose, local injection site reactions were reported by 67,521 (49.1%) BNT162b2 (Pfizer-BioNTech) and 92,472 (62.1%) mRNA-1273 (Moderna) vaccine recipients; systemic reactions were reported by 60,705 (44.2%) Pfizer-BioNTech and 76,756 (51.5%) Moderna vaccine recipients . Both loAmong 248,887 (86.9%) v-safe registrants aged \u226550 years who received homologous vaccination and a second mRNA booster dose, local injection site reactions were less frequently reported after the second booster dose than after any previous doses (p<0.001); systemic reactions were less frequently reported after the second booster than after either dose 2 or the first booster dose (p<0.001) . InabiliDuring March 29\u2013July 10, 2022, VAERS received and processed 8,515 reports of one or more adverse events after receipt of a second mRNA booster dose among adults aged \u226550 years . The medAmong the 8,515 VAERS reports of adverse events after receipt of a second mRNA booster dose among adults aged \u226550 years, 12 were preliminary reports of myocarditis (six nonserious and six serious); one case was verified by medical record review and met the CDC case definition for myocarditis; the patient was continuing to recover at time of this report.Among the 442 (5.2%) reports of serious events after second mRNA booster vaccination among adults aged \u226550 years 52 were reports of death; the median age of decedents was 84 years. For the six reports of death with sufficient information, cause of death as stated on the death certificate included congestive heart failure, aortic dissection, grand mal seizure, end-stage dementia, and cardiac arrest secondary to coronary artery disease. Among the serious events reported, 84 were of COVID-19 (19.0%).Limited data are available regarding the safety of second COVID-19 booster doses. The findings in this report are consistent with those from a small open-label clinical study of second boosters (154 participants received Pfizer-BioNTech and 120 Moderna vaccines) that did not detect any unexpected safety concerns . Preliminary safety findings after receipt of second booster doses among adults aged \u226550 years are similar to those after receipt of first booster doses. Reports of reactions after the second booster dose are less common than are those after the first. Health care providers and patients should be advised that local and systemic reactions are expected after second booster doses and that serious adverse events are infrequently reported. CDC and FDA will continue to monitor vaccine safety and will provide updates as needed to guide COVID-19 vaccination recommendations.ACIP recommends that all persons aged \u22655 years receive one booster dose of a COVID-19 vaccine after completion of their primary series; adults aged \u226550 years and persons aged \u226512 years with moderate to severe immunocompromise might receive a second booster \u22654 months after their first booster dose. Among adults aged \u226550 years, vaccine effectiveness against COVID-19-associated hospitalization \u2265120 days after dose 3 was 55% and \u22657 days after dose 4 was 80%, reinforcing recommendations that persons in this age group should receive a second booster when they become eligible (During March 29\u2013July 10, 2022, approximately 16.8 million persons in the United States aged \u226550 years received a fourth dose of a COVID-19 vaccine.Among persons aged \u226550 years who reported homologous mRNA COVID-19 vaccination, injection site and systemic reactions were less frequent after a second booster dose than after the first booster dose. Ninety-five percent of 8,515 events reported to the Vaccine Adverse Event Reporting System were nonserious.Health care providers and patients should be aware that local and systemic reactions are expected after a second mRNA COVID-19 booster dose. Serious adverse events are uncommon."} +{"text": "D3Cr muscle mass, grip strength, leg extension 1-rm max (Keiser), and walking speed were collected with standardized protocols. We calculated sex-stratified unadjusted Pearson correlation coefficients, and partial Pearson correlation coefficient adjusted for body size . D3Cr muscle mass was positively correlated with grip strength and leg extension strength . D3Cr muscle mass was correlated with 400-m walk speed only after adjustment for body size among both men and women . Associations of total thigh muscle volume by MRI with strength and walking speed were of similar magnitude to the association between D3Cr muscle mass with strength and walking speed."} +{"text": "Research has shown that emotional suppression, a form of emotion regulation, is often used by individuals with disordered eating behaviour. Moreover, eating disorder symptomatology is associated with inappropriate eating behaviours .The objective of the present study was to investigate the differences in eating behaviour among adults with different levels of emotional suppression and eating disorder symptomatology.Mage = 29.44 \u00b1 9.32) completed the Three-Factor Eating Questionnaire (eating behaviour), the Eating Attitudes Test (eating disorder symptomatology) and the Emotion Regulation Questionnaire .Two hundred seventy adults (N = 115) presenting low emotional suppression and low eating disorder symptomatology; cluster 2 (N = 43) presenting high emotional suppression and high eating disorder symptomatology and cluster 3 (N = 112) presenting high emotional suppression and low eating disorder symptomatology. Our results showed that individuals in cluster 2 had significantly greater levels of cognitive restraint, uncontrolled eating and emotional eating than individuals in clusters 1 and 3. Moreover, individuals in clusters 1 and 3 did not differ significantly in terms of any of the TFEQ subscales.Three clusters were identified through cluster analysis: cluster 1 (These preliminary findings may suggest that the tendency to persistently suppress emotions exacerbate disordered eating behaviour. Therefore, this factor together with symptoms of eating disorders should to be considered when planning prevention and intervention programs among adults presenting disordered eating behaviour.No significant relationships."} +{"text": "Displaced refugee children with a history of war exposure are at risk of developing complex and severe forms of post-traumatic stress disorder (PTSD).Search for the most relevant causal predictors of complex PTSD in a prospective cohort of Syrian refugee children living in informal settlements in Lebanon (N=1007).A latent class unsupervised analysis was carried out to determine clusters with complex PTSD presentation at the follow-up assessment. A new exploratory causal discovering modelling approach was applied using 97 multilevel psychosocial variables as predictors . Associations between discovered candidate causal factors assessed at baseline with a presumed diagnosis of complex PTSD one year later were calculated using a multiple logistic regression model.Several putative causal factors emerged: perceived social coherence of the neighbourhood ; impulsivity (1.25), self-efficacy (1.23) and depressive symptoms (1.15) at the parental level; positive home experiences (1.16) at the family level; and child-level factors such as being forced to work (1.22), being a victim of verbal or physical bullying (1.19), loneliness (1.17) and well-being (1.18). In further confirmatory multiple logistic regression analysis and after correction for multiple comparisons, verbal or physical bullying victimization (p=.005) and caregiver depressive symptoms (p=.0004) at baseline were associated with complex PTSD presentations one year later.Our results support the need for a multi-level psychosocial care model to prevent psychological distress and promote mental health in refugee children. Specifically, our results suggest that programs tackling caregiver\u2019s mental health and children\u2019s exposure to violence might effectively prevent complex PTSD.No significant relationships."} +{"text": "Djaalinj Waakinj cohort study (2017\u20132021),Otitis media and associated hearing loss are highly prevalent in Indigenous Australian children.Djaalinj Waakinj methodology has been described in detail elsewhere.The Djaalinj Waakinj study was approved by the Western Australian Aboriginal Human Ethics Committee (WAAHEC #759) and the Child and Adolescent Health Services Human Ethics Research Committee (CAHS HREC #12).The presence of middle ear effusion was assessed using tympanometry. Findings were classified by an audiologist: type A tympanograms were deemed to indicate normal middle ear function (no otitis media), type B tympanograms probable otitis media, and type C tympanograms probable Eustachian tube dysfunction. The Sixty\u2010seven of the 125 enrolled infants completed formal hearing assessments ; 41 were boys (61%). Twenty\u2010one of the 67 infants (31%) had normal hearing, 46 (69%) had some degree of hearing loss , including 22 with mild (33%) and 24 with moderate hearing loss (36%). Mean hearing responses were poorer for the 35 infants with abnormal tympanograms at the time of hearing assessment than for the 30 infants with normal tympanograms .Box 1* Normal: type A tympanogram (30 infants); abnormal: type B (30 infants) or type C (five infants). Tympanograms were not available for two infants. Visual reinforcement audiometry was performed with GSI 61 (Grason\u2013Stadler), Equinox (Interacoustics), and Avant (MedRx) audiometers, using pure tone, warble, or filtered narrowband noise stimuli through loudspeakers one metre from the child\u2019s ear at an angle of 90\u00b0. Tympanometry was conducted using either Titan Middle Ear Analyser (Interacoustics) or GSI 39 or MI 44 tympanometers (Maico).Our finding that 69% of urban Aboriginal infants in our study had hearing loss at about twelve months of age is comparable with that of a similar study in Kalgoorlie (65%).The mean hearing response in infants with abnormal tympanograms was 40.7\u2009dB, a level at which they would not hear normal voices clearly, with implications for later speech, language, and behavioural development.Open access publishing facilitated by The University of Western Australia, as part of the Wiley \u2010 The University of Western Australia agreement via the Council of Australian University Librarians.No relevant disclosures."} +{"text": "In the original version of this article, the allocation of the colours in the legend of Figure 1 was reversed. The correct legend should read as below and has now been corrected in the full article online.Figure 1: Computerized tomographic scan demonstrating anterior mediastinal soft tissue (arrow) in the (A) axial and (B) coronal planes. Axial T1W spin-echo magnetic resonance imaging scan (C) demonstrating anterior mediastinal soft tissue mildly hyperintense (yellow) compared to skeletal muscle control (red)."} +{"text": "Staphylococcus aureus (MRSA) and vancomycin-resistant Enteroccocus (VRE). Contezolid acefosamil is an intravenous (IV) double prodrug of CZD. Nonclinical and initial clinical data indicate CZA and CZD may cause less myelosuppression, particularly with longer duration therapy, and with reduced risk of monoamine oxidase inhibition compared with linezolid (LZD). In 3 CZD Phase 2 (Ph2) and Phase 3 (Ph3) skin infection trials and 1 CZA Ph2 acute bacterial skin and skin structure infection (ABSSSI) study, primary efficacy and overall safety outcomes were comparable to LZD, and the most common treatment emergent adverse events (TEAEs) were gastrointestinal; however, hematologic laboratory abnormalities and TEAEs were less common with CZD and CZA. In June 2021, CZD was approved in China for complicated skin and soft tissue infections (cSSTI). Sequential therapy with CZA IV followed by CZD PO is being evaluated in global Ph3 diabetic foot infection (DFI) and ABSSSI clinical trials. Because patients with diabetes commonly have diminished kidney function, efficacy and safety outcomes in subjects with renal impairment were evaluated in completed Ph2 and Ph3 CZD and CZA studies.Contezolid is a novel oral (PO) oxazolidinone with potent activity against Gram-positive pathogens, including methicillin-resistant In 4 CZD and CZA Ph2 and Ph3 skin infection trials, subjects were included with estimated creatinine clearance (CLcr) of 60 to < 90 mL/min (mild impairment) and 30 to < 60 mL/min (moderate impairment); no dose adjustments were made for renal function status. Primary efficacy outcomes and occurrence of TEAEs were evaluated for CZD and CZA subjects with no (CLcr \u226590 mL/min), mild, and moderate renal impairment.Primary efficacy outcomes and occurrence of TEAEs for CZD and CZA subjects with mild or moderate renal impairment were similar to that of subjects with no impairment in 4 skin infection trials.In 4 completed Ph2 and Ph3 skin infection clinical trials, subjects who received CZD or CZA with mild and moderate renal impairment appeared to have primary efficacy and safety outcomes similar to subjects with no impairment, supporting current Ph3 global DFI and ABSSSI studies which will enroll subjects with reduced kidney function.Edward Fang, MD, MicuRx Pharmaceuticals Inc: Employee Huahui Yang, MS, MicuRx Pharmaceuticals Inc: Employee."} +{"text": "This article has been corrected at the request of the author due to presence of an error in the Discussion section which was noticed by a reader. The second sentence of the first paragraph in the Discussion section has been corrected as the original version incorrectly referred to \"side effects\", when in reality they are \"clinical features\".Original (incorrect):\u00a0The common side effects of carbimazole include fever (92%), sore throat (85%), painful mouth ulcer (15%),\u00a0ulcer (8%), and reduced immune response making individuals prone to infections.Corrected:\u00a0The common clinical features of carbimazole induced agranulocytosis include fever (92%), sore throat (85%), painful mouth ulcer (15%),\u00a0ulcer (8%), and reduced immune response making individuals prone to infections."} +{"text": "Endolysins are produced by (bacterio)phages to rapidly degrade the bacterial cell wall and release new viral particles. Despite sharing a common function, endolysins present in phages that infect a specific bacterial species can be highly diverse and vary in types, number, and organization of their catalytic and cell wall binding domains. While much is now known about the biochemistry of phage endolysins, far less is known about the implication of their diversity on phage\u2013host adaptation and evolution. Using CRISPR-Cas9 genome editing, we could genetically exchange a subset of different endolysin genes into distinct lactococcal phage genomes. Regardless of the type and biochemical properties of these endolysins, fitness costs associated to their genetic exchange were marginal if both recipient and donor phages were infecting the same bacterial strain, but gradually increased when taking place between phage that infect different strains or bacterial species. From an evolutionary perspective, we observed that endolysins could be naturally exchanged by homologous recombination between phages coinfecting a same bacterial strain. Furthermore, phage endolysins could adapt to their new phage/host environment by acquiring adaptative mutations. These observations highlight the remarkable ability of phage lytic systems to recombine and adapt and, therefore, explain their large diversity and mosaicism. It also indicates that evolution should be considered to act on functional modules rather than on bacteriophages themselves. Furthermore, the extensive degree of evolvability observed for phage endolysins offers new perspectives for their engineering as antimicrobial agents. Endolysins are produced by bacteriophages to degrade the host cell wall and release new particles, but the implications of their diversity on phage-host adaptation and evolution is unknown. This study uses CRISPR-Cas9 genome editing to reveal novel insights into bacteriophage endolysin diversity and phage-bacteria interactions as well as into endolysin adaptation towards a new bacterial host. Bacteriophages (phages) exhibit exceptional structural and genetic diversity . A key fFor most dsDNA phages, host lysis at the end of the replication cycle is due to the coordinated actions of 2 proteins. Holins are proteins that control the timing of lysis by permeabilizing the inner membrane of the host to allow the diffusion of the lytic enzymes, namely, the endolysins. The latter then gains access and degrades the cell wall peptidoglycan to induce lysis . DestabiN-acetylglucosamine (GlcNAc)\u2013N-acetylmuramic acid (MurNAc) glycan strands that are cross-linked by short stem peptides attached to MurNac residues . TM. TM43]. (TIF)Click here for additional data file.S5 FigProteins were loaded on NuPAGE 4%\u201312% BisTris gels and stained with Coomassie Blue. The expected molecular mass of each purified protein is indicated on the top of the figure. Molecular weight markers are on the left.(TIF)Click here for additional data file.S6 Fig(A) Schematic representation of the LysP008 endolysins domains and its alternative start codon (M 172) observed at the beginning of EME_EF1-like CBD. The 25-bp present before the alternative start codon (in blue) compared to a consensus sequence generated with Weblogo [ Weblogo and 14 r(TIF)Click here for additional data file.S7 Fig(A) Alignment of the 25-bp present before the internal start codon of the LysP008 and Lysc2 genes with or without codon optimization for E. coli BL21. (B) Purification of the LysP008 and Lysc2 without codon optimization and size exclusion chromatography (2 liters cell culture).(TIF)Click here for additional data file.S8 Fig(A) Analysis of non-lactococcal phages that have endolysins with at least 90% coverage and 50% identity to endolysins of lactococcal phages (L. lactis phages are grouped according to their type of endolysin (l phages . B) Analndolysin . CD, cat(TIF)Click here for additional data file.S9 FigE. faecalis. ClustalW (v2.1) was used to perform multiple alignments and generate a phylogenetic tree (We investigated the diversity of endolysins found in 70 complete genomes of phages that infect tic tree . Conserv(TIF)Click here for additional data file.S10 FigL. fermentum. ClustalW (v2.1) was used to perform multiple alignments and generate a phylogenetic tree (We investigated the diversity of endolysins found in 3 complete genomes from phages infecting tic tree . Conserv(TIF)Click here for additional data file.S11 FigS. themophilus. ClustalW (v2.1) was used to perform multiple alignments and generate a phylogenetic tree (We investigated the diversity of endolysins found in 56 complete genomes from phages infecting tic tree . Conserv(TIF)Click here for additional data file.S12 FigRecombination between the prophage bIL285 and the virulent phage P335 was observed after either 10 or 20 transfers. A cutoff of 95% identity was used for the genome\u2019s alignment. Homology regions were recombination between the phage P335 and prophage BIL285 take place are highlighted in red.(TIF)Click here for additional data file.S13 Fig(A) Image of the phage P335 plaques at a resolution of 31 pixels per mm. (B) Using ImageJ [g ImageJ , lytic p(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S1 Data(XLSX)Click here for additional data file.S2 Data(XLSX)Click here for additional data file.S3 Data(XLSX)Click here for additional data file.S4 Data(XLSX)Click here for additional data file.S5 Data(XLSX)Click here for additional data file.S6 Data(XLSX)Click here for additional data file.S7 Data(XLSX)Click here for additional data file."} +{"text": "Background: Operation room (OR) nurses are at a high risk for bloodborne pathogen (BBP) exposures because they are constantly in contact with blood and body fluid (BBF) through surgical sites, and they frequently use sharp surgical instruments and sutures during surgeries. We explored occupational experiences with BBP exposures among OR nurses in South Korea. Methods: With an institutional review board\u2019s approval, this qualitative research was performed with 12 OR nurses who had worked for >3 months and had experienced BBP exposures. Based on the main research question \u201cHow is the experience of BBP exposure among OR nurses?\u201d the semistructured questions for in-depth interview were prepared. Narrative data were collected through 1-hour individual interviews from June to September 2020 and were analyzed using a thematical analysis method. Results: The average age of the participants was 34.4 years with an average 9.75 years of clinical experience. The main theme extracted was \u201cThe nurses are alerted to their safety after experiencing the aftereffects and emotional trauma from BBP exposures,\u201d with 4 subthemes and 14 concepts. The first subtheme, \u201cOR nurses risking exposure to BBF,\u201d included (1) hurried doctors and hasty nurses; (2) sharp surgical instruments everywhere; (3) deprioritized self-protection due to ongoing surgery; (4) inattentive to BBF risk; and (5) uncomfortable, foggy goggles and receded safety devices. The second subtheme, \u201cBBP exposures occurred in a flash,\u201d included (1) sharp injury occurred in a split second; (2) temporarily treat sharp wounds while hiding frightened feelings; and (3) OR nurses concentrated on surgery by suppressing anxiety and sharp wounds. The third subtheme, \u201cBurdened time that could be overwhelmed alone,\u201d included (1) BBP exposures moments that I wish to reverse; (2) anger over dangerous environments and the turmoil of anxiety; (3) exhausted body and facial discoloration due to taking postexposure prophylaxis; and (4) exposure to BBP that I want to hide from family and friends. The fourth subtheme, \u201cVoice for everyone\u2019s safety,\u201d included (1) establishing a safety culture, which requires everyone\u2019s efforts, and (2) necessity of practical resources for decreasing BBP exposures. Conclusions: The participating OR nurses felt that they were working with a high risk to BBF exposures and called for institutional interventions to reduce the risks, including surgeons\u2019 attentive collaboration. Korean hospitals should make greater efforts to establish safety culture in ORs and to provide repeated, tailored education to prevent BBF exposures. They should also supply high-quality protective equipment and safety-engineered devices for OR nurses.Funding: NoDisclosures: None"} +{"text": "The objective was to examine the relationship between maternal intimate partner violence (IPV) victimization and children\u2019s developmental health using linked population-wide administrative datasets. We examined developmental vulnerability (DV) at kindergarten of children exposed to maternal IPV victimization aged 0 to 5 using provincial prosecution records compared to unexposed counterparts.This retrospective cohort study linked administrative datasets from the Population Research Data Repository at the Manitoba Centre for Health Policy. Exposed mother-child pairs with 1+ prosecution records of maternal IPV victimization during early childhood (child aged 0 to 5) between 2003-2018 in Manitoba were matched to unexposed pairs (1:3) based on sex/birthdate of child and neighbourhood income. DV at kindergarten was measured across 5 domains using the Early Developmental Instrument (EDI). Children without eligible EDI scores were excluded. Multiple logistic regression models were conducted.The cohort included 5321 children . 32.98% of the cohort was developmentally vulnerable in one or more domains (1/+) and 19.60% was developmentally vulnerable in two or more domains (2/+). Unadjusted relationships between maternal IPV victimization from age 0 to 5 and developmental vulnerability at kindergarten were statistically significant across all 5 domains and in 1/+ as well as 2/+ domains. After adjusting for covariates, children who were exposed to maternal IPV victimization from ages 0 to 5 had increased odds of being developmentally vulnerable in social competence and emotional maturity , also in 2/+ domains at kindergarten, compared to unexposed counterparts.The study provided Canadian population-wide evidence of the association between maternal IPV victimization and early childhood development, specifically later socio-emotional vulnerability. Interventions and support systems for this population of families should be developed and implemented, with an emphasis on mitigating long-term socio-emotional developmental risks in children exposed to IPV."} +{"text": "In Bolton Early Intervention Team (EIT) it was noticed that patients prescribed antipsychotics frequently required a change in medication due to side effects. Similar issues had been identified in Avon and Wiltshire NHS Foundation Trust where a prescribing guideline was developed which won the NICE Shared Learning Award in 2020. This recommends prescribing Aripiprazole first line and cautions using Olanzapine or typical antipsychotics first due to their side effects. The aim of this project was to identify which antipsychotic drugs are currently prescribed in first episode of psychosis (FEP) in Bolton EIT patients and to audit adherence to National Institute of Clinical Excellence antipsychotic prescribing guideline CG178.The sample included all adults with FEP accepted by Bolton EIT across a four-month period from 01/12/20 until 31/03/21. Fifty-two people were identified.Measured standards were documentation of prescribing rationale, discussion regarding medication side effects and weekly weight monitoring for six weeks following initiation. Antipsychotic choice and need for a change within six months of initiation was recorded. Data were collected retrospectively from patients\u2019 electronic records.Thirty-eight patients had been prescribed an antipsychotic \u2013 fifteen as inpatients, seventeen by Bolton EIT and six by the Home Treatment Team.Of the fifteen inpatients Olanzapine (8) and Zuclopenthixol (3) were the most common choice. 5/15 had a documented rationale, and side effects were discussed with 3/15 patients. Weekly weight monitoring was performed in 7/15.Of the 17 people who started antipsychotic medication once under Bolton EIT Quetiapine (6), Olanzapine (6) and Aripiprazole (5) were the most common choices. 12/17 had a documented rationale and 13/17 were consulted regarding side effects. Weekly weight monitoring was not performed for any of these patients.Within six months, sixteen antipsychotic prescriptions (42%) were changed due to side effects (9), inefficacy (6) and non-compliance (1). The drugs changed were Olanzapine (6) Quetiapine (6) Zuclopenthixol (2) Aripiprazole (1) and Chlorpromazine (1).Those initiated on antipsychotics as inpatients need better involvement in decision-making and consultation about side effects. A community initiative should be introduced to offer weekly weight monitoring. Further work is required to understand the rationale for frequently prescribing Olanzapine and Zuclopenthixol in inpatient services, and to consider why Aripiprazole is infrequently used first line."} +{"text": "Nature Communications 10.1038/s41467-022-31388-z, published online 24 June 2022Correction to: The original version of this Article contained an error in the section of the Results, which incorrectly read \u2018.\u2019 The correct version states \u2018\u2019.n = 3). j. Representative Immunofluorescence staining images of VWF and TAGLN in adipose tissues. Arrow (red) indicates ECs expressing both VWF and TAGLN. Arrow (green) indicates ECs only expressing VWF (n = 3). k. IHC of FABP4 in ECs and adipose tissues. Arrows (red) indicate ECs. Arrows (green) indicate adipocytes. Control was stained with an antibody against a gene not expressed in adipose tissues (n = 3).>.\u2019 The correct version states \u2018\u2019.The original version of this Article contained an error in the legend to Figure 3, which incorrectly read \u2018.\u2019 The correct version states \u2018\u2019.This has been corrected in both the PDF and HTML versions of the Article."} +{"text": "This case series assesses the diseases misdiagnosed as autoimmune encephalitis and potential reasons for misdiagnosis. What diseases are misdiagnosed as autoimmune encephalitis and which factors contribute to misdiagnosis?In this case series of 107 outpatients misdiagnosed with autoimmune encephalitis, approximately half had functional neurologic or psychiatric disorders. An insidious rather than subacute onset and lack of magnetic resonance imaging or cerebrospinal fluid findings suggestive of inflammation were clues to misdiagnosis; overinterpretation of serum nonspecific antibodies was a major contributor to misdiagnosis.A broad range of disorders are misdiagnosed as autoimmune encephalitis and misdiagnosis occurs in many settings including at specialized centers participating in this study. Autoimmune encephalitis misdiagnosis can lead to harm.To determine the diseases misdiagnosed as autoimmune encephalitis and potential reasons for misdiagnosis.This retrospective multicenter study took place from January 1, 2014, to December 31, 2020, at autoimmune encephalitis subspecialty outpatient clinics including Mayo Clinic (n\u2009=\u200944), University of Oxford (n\u2009=\u200918), University of Texas Southwestern (n\u2009=\u200918), University of California, San Francisco (n\u2009=\u200917), University of Washington in St Louis (n\u2009=\u20096), and University of Utah (n\u2009=\u20094). Inclusion criteria were adults (age \u226518 years) with a prior autoimmune encephalitis diagnosis at a participating center or other medical facility and a subsequent alternative diagnosis at a participating center. A total of 393 patients were referred with an autoimmune encephalitis diagnosis, and of those, 286 patients with true autoimmune encephalitis were excluded.Data were collected on clinical features, investigations, fulfillment of autoimmune encephalitis criteria, alternative diagnoses, potential contributors to misdiagnosis, and immunotherapy adverse reactions.N-methyl-d-aspartate receptor by cell-based assay only , and other (n\u2009=\u200918). Adverse reactions from immunotherapies occurred in 17 of 84 patients (20%). Potential contributors to misdiagnosis included overinterpretation of positive serum antibodies (53 [50%]), misinterpretation of functional/psychiatric, or nonspecific cognitive dysfunction as encephalopathy (41 [38%]).A total of 107 patients were misdiagnosed with autoimmune encephalitis, and 77 (72%) did not fulfill diagnostic criteria for autoimmune encephalitis. The median (IQR) age was 48 (35.5-60.5) years and 65 (61%) were female. Correct diagnoses included functional neurologic disorder (27 [25%]), neurodegenerative disease (22 [20.5%]), primary psychiatric disease (19 [18%]), cognitive deficits from comorbidities (11 [10%]), cerebral neoplasm (10 [9.5%]), and other (18 [17%]). Onset was acute/subacute in 56 (52%) or insidious (>3 months) in 51 (48%). Magnetic resonance imaging of the brain was suggestive of encephalitis in 19 of 104 patients (18%) and cerebrospinal fluid (CSF) pleocytosis occurred in 16 of 84 patients (19%). Thyroid peroxidase antibodies were elevated in 24 of 62 patients (39%). Positive neural autoantibodies were more frequent in serum than CSF (48 of 105 [46%] vs 7 of 91 [8%]) and included 1 or more of GAD65 (n\u2009=\u200914), voltage-gated potassium channel complex (LGI1 and CASPR2 negative) (n\u2009=\u200910), When evaluating for autoimmune encephalitis, a broad differential diagnosis should be considered and misdiagnosis occurs in many settings including at specialized centers. In this study, red flags suggesting alternative diagnoses included an insidious onset, positive nonspecific serum antibody, and failure to fulfill autoimmune encephalitis diagnostic criteria. Autoimmune encephalitis misdiagnosis leads to morbidity from unnecessary immunotherapies and delayed treatment of the correct diagnosis. Yet, there are limited data concerning patients initially incorrectly diagnosed with autoimmune encephalitis and their subsequent correct diagnosis. This is an important topic given the risk of patient harm associated with misdiagnosis, including morbidity from adverse effects of immunotherapies and delay of appropriate treatment.8 We report data from an international multicenter study of autoimmune encephalitis misdiagnosis across 6 subspecialty centers to analyze patients misdiagnosed with autoimmune encephalitis and identify possible contributors to misdiagnosis.Autoimmune encephalitis is increasingly a diagnostic consideration in patients with subacute onset of memory loss, altered mental status, and/or psychiatric symptoms\u2014core features of proposed diagnostic criteria.STROBE) reporting guideline for reporting observational studies.The Mayo Clinic institutional review board approved this multicenter study (#19-004926), and institutional review board approval also occurred at each respective site with all patients either providing written consent or patients included under an institutional review board approved consent waiver for minimal risk retrospective studies. This study was a retrospective multicenter observational study that followed the Strengthening the Reporting of Observational Studies in Epidemiology (Inclusion criteria were adult patients (18 years or older) at the time of neurologic evaluation at a participating site with (1) a prior autoimmune encephalitis diagnosis assigned at another medical center or at the participating site and occurring in the inpatient or outpatient setting and (2) a subsequent alternative diagnosis made at an in-person visit at one of the participating outpatient autoimmune neurology clinics. Alternative diagnoses were defined as a definite alternative diagnosis when diagnostic testing confirmed the diagnosis or as a clinical alternative diagnosis when definitive confirmation was not available or it was a purely clinical diagnosis .10 At the University of California San Fransisco, only patients who received immunotherapy for their presumed autoimmune encephalitis diagnosis were included. Details on numbers of true autoimmune encephalitis over the same study time frame, when available, were also collected to assess its frequency.Six academic medical centers with subspecialty expertise in autoimmune neurology participated. These included Mayo Clinic in Rochester, Minnesota (autoimmune neurology clinic); University of Oxford in Oxford, United Kingdom (autoimmune neurology clinic); University of Texas Southwestern in Dallas (autoimmune neurology clinic); University of California, San Francisco in San Francisco ; Washington University in St Louis in St Louis, Missouri ; and University of Utah in Salt Lake City (autoimmune neurology clinic). Patients evaluated clinically between January 1, 2014, to December 31, 2020, were considered for study enrollment. Data on 2 patients included in the study were previously published in case reports.1 (1) subacute onset (rapid progression of <3 months) of working memory deficits (short-term memory loss), altered mental status, or psychiatric symptoms and (2) at least one of the following: new focal central nervous system findings, seizures not explained by a previously known seizure disorder, cerebrospinal fluid (CSF) pleocytosis (white blood cell count of >5 cells/mm3), or magnetic resonance imaging (MRI) brain features of encephalitis with either hyperintense signal on T2-weighted fluid-attenuated inversion recovery sequences highly restricted to 1 or both medial temporal lobes or in multifocal areas involving gray matter, white matter, or both compatible with demyelination or inflammation.Participating centers provided deidentified data detailing age, sex, clinical, and paraclinical variables from patients misdiagnosed with autoimmune encephalitis. Data on race and ethnicity were not collected. Data on the requirements for part 1 and part 2 of the diagnostic criteria for possible autoimmune encephalitis were also specifically collected and includeFailure to fulfill both part 1 and 2 of the criteria precludes a diagnosis of any category of autoimmune encephalitis. Part 3 of the autoimmune encephalitis diagnostic criteria was not analyzed as this component specifies reasonable exclusion of alternative diagnoses, which by design of the present study would be difficult to quantify retrospectively.Data collected included age at symptom onset, sex, and time from disease onset to correct diagnosis, insidious (symptoms developing over \u22653 months) vs subacute (<3 months) onset, cancer history, thyroid autoimmunity, or other autoimmune disorders. Results of neuropsychological testing were classified as normal (for age and education) or abnormal. We collected data on elevated IgG index, CSF-restricted oligoclonal bands, electroencephalogram , thyroid peroxidase antibodies, other serologic evidence of systemic autoimmunity, and serum and CSF anti-neural or glial antibodies (including information on titer and assay type when available). Brain biopsy or autopsy details were obtained when applicable. Information on immunotherapies and adverse reactions were also collected.Participating sites selected from the following potential reasons for misdiagnosis in each patient: (1) overinterpretation of a nonspecific positive antibody; (2) failure to accept an alternative psychiatric diagnosis; (3) misclassification of functional neurologic symptoms as true neurologic abnormalities; (4) overinterpretation of nonspecific cognitive symptoms as encephalitis; or (5) other. There was also a free text section for additional reasons for misdiagnosis.Descriptive statistics were used. For categorical variables, frequency and percent were used, whereas for continuous variables, median and range or interquartile range were used. JMP Pro, version 14.1.0 was used.We included 107 patients misdiagnosed as having autoimmune encephalitis at the 6 participating centers. The median (IQR) age at symptom onset was 48 (35.5-60.5) years and 65 (61%) were female. The median (IQR) time from onset to the correct diagnosis was 16 (7-40) months. A history of any type of autoimmune disease was noted in 44 individuals (41%), of whom 34 (77%) had thyroid autoimmunity. Six patients (6%) had a history of cancer outside of the nervous system. Symptom onset was insidious in 51 of 107 patients (48%), although some had superimposed subacute worsening.Autoimmune encephalitis misdiagnosis occurred in 107 individuals during a period over which 286 were correctly diagnosed as having autoimmune encephalitis. This included Mayo Clinic ; University of Oxford ; University of Texas Southwestern ; University of California, San Francisco ; Washington University in St Louis ; and University of Utah .Alternative diagnoses are detailed in Those fulfilling part 1 of the criteria had 1 or more of a clinical presentation of a subacute onset (rapid progression of <3 months) with 1 or more of working memory deficits (short-term memory loss) (36 [34%]), altered mental status (43 [40%]), or psychiatric symptoms (42 [39%]).Those fulfilling part 2 of the criteria had 1 or more of the following: (1) focal central nervous system findings in 31 patients (29%); (2) seizures not explained by a previously known seizure disorder in 26 patients (24%); (3) CSF pleocytosis in 16 of 84 patients (19%); or (4) MRI brain features suggestive of encephalitis in 19 of 104 patients (18%) with either features of limbic encephalitis in 10 A or mulIn total, 77 patients (72%) did not fulfill autoimmune encephalitis diagnostic criteria as they lacked requirements for possible autoimmune encephalitis diagnosis, which is a prerequisite for any other autoimmune encephalitis diagnostic category.Thyroid peroxidase antibodies were positive in 24 of 62 individuals (39%). Nineteen patients had coexisting serologic evidence of systemic autoimmunity with antinuclear antibody positivity most common. Neural autoantibodies were identified more often in serum (48 of 105 [46%]) than CSF (7 of 91 [8%]) and are outlined in Neuropsychological test results were abnormal in 38 of 54 patients (70%). Electroencephalogram findings were abnormal in 31 of 79 (39%) and revealed epileptiform abnormalities in 16 and slowing in 9; details of abnormalities were not available in 6 patients. CSF-restricted oligoclonal bandings or IgG index positivity occurred in 7 of 82 (9%) tested.N-methyl-d-aspartate receptor (NMDAR) antibodies in the CSF without evidence on mouse tissue-based indirect immunofluorescence had HIV-associated leukoencephalopathy with treatment-related adverse reactions documented in 17 of 84 patients (20%) .The reasons for misdiagnosis included 1 or more of overinterpretation of a nonspecific positive antibody result (53 [50%]); misinterpretation of nonspecific symptoms as neurologic (19 [18%]); imaging findings felt to be consistent with autoimmune encephalitis (15 [14%]); functional neurologic features mistaken for true neurologic symptoms (14 [13%]); abnormal cerebrospinal fluid findings (9 [8%]); psychiatric manifestations thought to be from autoimmune encephalitis (8 [7%]); failure to accept a psychiatric diagnosis (5 [5%]); or subacute onset or fluctuating course (4 [4%]).14This study highlights that misdiagnosis of autoimmune encephalitis is an important and frequent clinical problem. Autoimmune encephalitis misdiagnosis was identified at participating subspecialty outpatient clinics, but the initial incorrect autoimmune encephalitis diagnosis occurred at both outside facilities and participating centers. This shows that misdiagnosis of autoimmune encephalitis can be encountered in multiple settings, including at autoimmune neurology subspeciality clinics with focused expertise. Many of these patients endured a delay to their correct diagnosis for longer than a year, and one-fifth experienced morbidity related to unnecessary immunotherapy. Overinterpretation of a nonspecific autoantibody was a frequent contributor to misdiagnosis. In 72% of patients, they did not fulfill autoimmune encephalitis diagnostic criteria, suggesting more stringent adherence to these criteria may prevent misdiagnoses. In particular, an insidious onset of symptoms and absence of MRI or CSF findings suggestive of neuroinflammation should raise suspicion for an alternative diagnosis. Yet, patients with LGI1 , CASPR2m and IgLON5 antibodies can present over long durations with minimal evidence of paraclinical investigation abnormalities, other than the autoantibody itself.16 This is similar to recent data concerning multiple sclerosis misdiagnosis.17 Functional neurologic disorders and psychiatric diseases are highly prevalent alternative diagnoses whose distinction from autoimmune encephalitis can be challenging.21 Autoimmune encephalitis is increasingly considered in patients with psychiatric symptoms as it is potentially treatable with immunotherapy, but autoimmune encephalitis is much less common than primary psychiatric disease, for instance, accounting for less than 1% presenting with a typical first episode of psychosis.23 Psychiatric disease combined with other contributors to cognitive deficits such as chronic pain, sleep disturbance, and medication adverse reactions also led to misdiagnosis. Such patients often had normal neuropsychological testing and did not fulfill autoimmune encephalitis diagnostic criteria due to absence of MRI and CSF findings suggesting classic neuroinflammation.Autoimmune encephalitis is a rare condition, with a cumulative incidence of\u2009approximately\u20093 to 9 per million person-years and common conditions accounted for a high proportion of cases mistaken for autoimmune encephalitis.24 Imaging and CSF analysis for amyloid and tau and CSF prion detection with real-time quaking-induced conversion are novel biomarkers that aid diagnosis of Alzheimer disease and Creutzfeldt-Jakob disease, respectively.26Neurodegenerative disorders accounted for 20% of misdiagnoses and the insidious onset and absence of neuroinflammation on testing help discriminate from autoimmune encephalitis. However, fluctuations in patients with Lewy body disease and rapid progression with overlapping MRI findings in Creutzfeldt-Jakob disease can make this distinction challenging.8 The multifocal MRI abnormalities, CSF pleocytosis, and steroid responsiveness of central nervous system lymphoma mimicked autoimmune encephalitis here and previously.27 The subacute encephalopathy, cortical swelling, and signal abnormality on MRI with mitochondrial encephalomyopathy lactic acidosis and strokelike episodes mimicked autoimmune encephalitis similar to prior reports.28 Seizure-related MRI signal abnormalities can overlap with autoimmune encephalitis MRI findings and lead to misdiagnosis.29 Thiamine deficiency and HIV infection are important treatable mimics identified here and reported previously.31 Taken together, the aforementioned cases pose a particular challenge given the paraclinical features in common with autoimmune encephalitis.We found 28% of patients fulfilled autoimmune encephalitis criteria and such patients usually had overlapping MRI or CSF findings with autoimmune encephalitis. Temporal lobe glioma may mimic autoimmune encephalitis; however, the absence of sustained response to immunotherapy, presence of mass effect on MRI , A and l32 With neural autoantibody biomarkers the diagnostic accuracy varies by pretest probability, sample assessed (serum or CSF), antibody type, assay methodology, and antibody titer.6 As up to 5% of patients may harbor a positive neuronal antibody, clinically irrelevant results may be frequent if many patients are serologically assessed.33 Indeed, in this study, some positives were misinterpreted as being relevant despite autoimmune encephalitis not being the typical phenotype, suggesting that removing problematic antibodies with low specificity from autoimmune encephalitis autoantibody panels could reduce misdiagnosis.37 Low-end titer serum GAD65 antibody positives were often overinterpreted as supporting autoimmune encephalitis but occur in 8% of the population and typically only high titer (>10\u2009000 IU/mL using enzyme-linked immunosorbent assay or >20 nmol/L using radioimmunoassay)39 serum positives or CSF detection are neurologically relevant.42 Laboratories offering serum GAD65 antibody testing for neurologic indications should consider using these higher cutoffs for neurologically relevant positivity. Voltage-gated potassium channel complex antibody positivity without LGI1 or CASPR2 reactivity are not useful for autoimmune encephalitis diagnosis,44 while low-titer CASPR2 antibodies are also problematic and only high titers support autoimmune encephalitis.47 Serum NMDAR antibodies with negative CSF results were a red flag here, as noted previously.48 Rarely, CSF NMDAR antibodies by cell-based assay alone led to misdiagnosis. Despite its high specificity, these positive results in CSF may relate to diffusion of high serum levels, rather than intrathecal synthesis. Detection using a second rodent tissue-based assay enhances CSF NMDAR antibody specificity further.48 Antibodies detected by western blot/line blot or immunoblot in isolation often yield false positives and require cautious interpretation.50 Moreover, detection of neural antibodies in noncertified laboratories require extreme caution. While this study focused only on autoimmune encephalitis, overinterpretation of nonspecific antibodies is also problematic in other neurologic syndromes in which antibodies are tested . Increased education of neurologists on when to order neural autoantibodies and how to interpret positive results is needed to reduce the risk of misdiagnosis and interpretative comments provided by laboratories reporting results can be helpful in this regard.52Overinterpretation of a nonspecific antibody was the largest potential contributor to autoimmune encephalitis misdiagnosis and a list of the more problematic antibodies are summarized in the Box. Insidious onsetMultiple comorbidities that cause cognitive deficits such as polypharmacy, chronic pain, fibromyalgia, sleep disordersExamination results consistent with functional neurologic disorderFeatures of mitochondrial disease presentNormal neuropsychological test resultsNormalProgressive atrophy without signal abnormalities or enhancementLesion(s) continuing to expand despite immunotherapyaNoninflammatoryTPO antibodies of any titerLow titer\u2013positive GAD65 antibodiesVoltage-gated potassium channel complex antibodies negative for LGI1/CASPR2Low-titer antibody positives by older generation techniques Isolated serum NMDAR antibody negative in CSFImmunoblot or line blot antibody positivity in isolationLow titer positive\u2013CASPR2 antibodiesAntibody detection in noncertified laboratoriesN-methyl-d-aspartate receptor; RIA, radioimmunoprecipitation assay; TPO, thyroid peroxidase.Abbreviations: CASPR2, contactin-associated protein-like 2; CSF, cerebrospinal fluid; GAD65, glutamic acid decarboxylase 65; LGI1, leucine-rich-glioma-inactivated-1; NMDAR, aNormal white blood cell count and absence of CSF unique oligoclonal bands.54 Finally, increased health care costs may arise from the use of expensive immunosuppressants or unnecessary evaluation for an underlying cancer prompted by nonspecific antibody detection.Autoimmune encephalitis misdiagnosis is problematic for multiple reasons. First, misdiagnosis of autoimmune encephalitis increases morbidity from failure to treat the actual diagnosis. Second, immunosuppressant treatments commonly have adverse reactions that may be serious, and in this study included infection, psychosis, avascular necrosis of the hip, and heart failure. Moreover, there are many less severe, yet common and bothersome, adverse reactions of corticosteroids including insomnia, weight gain and irritability, some of which may not have been captured in this analysis. Third, during the COVID-19 pandemic, immunotherapies may increase risk of severe COVID-19 infection and hinder vaccine and natural infection responses.1 The selection bias of analyzing autoimmune encephalitis misdiagnosis identified at subspecialty autoimmune neurology clinics could underestimate the rate of autoimmune encephalitis misdiagnosis and it may exceed true autoimmune encephalitis diagnosis in the general population. There are many potential contributors to underrepresentation of autoimmune encephalitis misdiagnosis including our requirement for an in-person visit as autoimmune encephalitis misdiagnosis can be identified in other settings . Moreover, during triage for appointments, true autoimmune encephalitis may be favored over cases suspected to be misdiagnosed. Also, infectious mimics of autoimmune encephalitis are more likely to be encountered in hospitalized patients and our study focused on those identified at outpatient clinics.55 Finally, differences in rates of autoimmune encephalitis misdiagnosis across centers likely reflect variation in referral patterns. Further studies are needed to better capture autoimmune encephalitis misdiagnosis rates across other settings.The retrospective design was a limitation and prospective studies are needed to assess autoimmune encephalitis misdiagnosis frequency and characteristics among new referrals to subspecialty clinics with presumed autoimmune encephalitis. Such studies could incorporate probable and definite categories of autoimmune encephalitis diagnostic criteria to better discriminate true autoimmune encephalitis from autoimmune encephalitis misdiagnosis.In summary, neurologists should be aware of the potential for autoimmune encephalitis misdiagnosis and consider a broad differential diagnosis including common disorders when evaluating suspected cases. Improved recognition of the clinical, imaging, and serologic red flags in the evaluation of autoimmune encephalitis summarized in the"} +{"text": "Vocal learning is thought to have evolved in 3 orders of birds , with each showing similar brain regions that have comparable gene expression specializations relative to the surrounding forebrain motor circuitry. Here, we searched for signatures of these same gene expression specializations in previously uncharacterized brains of 7 assumed vocal non-learning bird lineages across the early branches of the avian family tree. Our findings using a conserved marker for the song system found little evidence of specializations in these taxa, except for woodpeckers. Instead, woodpeckers possessed forebrain regions that were anatomically similar to the pallial song nuclei of vocal learning birds. Field studies of free-living downy woodpeckers revealed that these brain nuclei showed increased expression of immediate early genes (IEGs) when males produce their iconic drum displays, the elaborate bill-hammering behavior that individuals use to compete for territories, much like birdsong. However, these specialized areas did not show increased IEG expression with vocalization or flight. We further confirmed that other woodpecker species contain these brain nuclei, suggesting that these brain regions are a common feature of the woodpecker brain. We therefore hypothesize that ancient forebrain nuclei for refined motor control may have given rise to not only the song control systems of vocal learning birds, but also the drumming system of woodpeckers. Vocal learning is thought to have evolved in three orders of birds . This study shows that woodpeckers have evolved a set of brain nuclei to mediate their drum displays, and these regions closely mirror those that underlie song learning in songbirds. Advanced vocal learning is a rare trait, which thus far has been found in only 3 of over 40 avian lineages and 5 of over 30 mammalian lineages . Among Fig 1A), much like birdsong in some species diluted in TNT buffer) for 1 h at room temperature. Next, slides were incubated in anti-DIG-HRP antibody diluted in TNT buffer overnight at 4\u00b0C. The following day, slides were washed 3 times in TNT buffer for 10 min each. We then incubated slides in FITC-TSA (1:150) diluted in amplification buffer (Akoya Biosciences) for 10 min. Finally, slides were washed 3 times in PBS and then mounted in Prolong Gold with DAPI.Brain sections were first fixed in 4% paraformaldehyde for 5 min and then washed in PBS twice for 3 min each. Sections were then acetylated for 10 min in a 0.1-M TEA solution with 0.33% acetic anhydride, rinsed once with PBS, and then serially dehydrated in 70%, 95%, and 100% ethanol. Sections were next incubated in hybridization at room temperature for 1 h. Slides were then transferred to hybridization buffer with antisense DIG and anterior nidopallium (AN), respectively). For the AI, we counted all the cells within a fixed region (500 \u00d7 500 \u03bcm square) that was directly below the PV-rich dorsal arcopallial region . To count cells in AN, we restricted our counts to the region that immediately surrounded dAN . For all 4 regions, PV cells counts were normalized to area in which cells were counted (number of PV+ cells/\u03bcm2).Colormetric in situ slides were visualized on a Ziess Axiozoom V16 stereoscope. All images for IEG quantification were taken at 32\u00d7 optical zoom using ZenBlue. The number of EGR1 and Arc, regions of interests were determined by aligning an adjacent PV image in Photoshop and then outlining the region of PV-rich cells. Two investigators who were blind to the condition of the birds counted the total number of Arc or EGR1 expressing cells in these regions of interest. The region we chose is consistent with detailed reports annotating the AI [2) of each brain region. Then, as in many previous analyses with IEG analyses in the song system [Arc and EGR1 mRNA expression. Fluorescent in situ hybridization for PV mRNA were visualized using a Zeiss LSM 710 confocal microscope using a 10\u00d7 objective. Tile scans of entire sections were acquired in the Zen software.To quantify both g the AI . For theg the AI . Finallyg the AI and IEG g the AI ,26. Cellg system ,32,43, wArc cell counts to achieve normality. Using Q\u2013Q plots and Shapiro-Wilk tests, we verified that these transformations did in fact yield a more normally distributed data. We used a one-way analysis of variance (ANOVA) to investigate whether Arc and EGR1 expression differed in the arcopallial regions (dA and AI), nidopallium (dAN and AN), and auditory regions (NCM and CMM) between drumming, vocalizing, silent, and passively behaving woodpeckers. For these analyses, the drumming condition consisted of birds that only drummed (n = 4) and those that drummed and produced a small number of whinny calls (n = 6). Since Arc expression levels did not differ between these 2 groups for either the dA or the dAN , we collapsed these into a single condition called \u201cdrumming\u201d. Linear regression analyses were performed to determine if Arc mRNA expression in the arcopallial or nidopallial regions were associated with the number of drums, vocalizations , aggressive flights, or total behavior produced during the STI. To investigate Arc \u201ccontrast\u201d in putative drum nuclei (dA and dAN), we took the ratio of normalized Arc cell counts (Arc+ cells/\u03bcm2) in dA or dAN to normalized cell counts in in AI or AN . To evaluate whether drumming birds had higher contrast relative to nondrumming birds, we performed an independent sample t test for each ratio.We performed analyses in R, after log (1+x) transforming normalized S1 FigPV mRNA in species of (A) Humbolt penguin (Spheniscus humboldti) and (B) emu (Dromaius novaehollandia). In contrast to hummingbirds and woodpeckers are as follows: Hyper, hyperpallium; Meso, mesopallium; Nido, nidopallium; GP, globus pallidus; T, Thalamus; Ot, optic tectum; St, striatum; Arco, arcopallium. Photo credits: penguin from Mariana Ruiz Villarreal (CC BY 2.0) and emu from Daderot (CC Public Domain via wikimedia).(A-B) Representative radioactive in situ hybridization microscope images of kers see , PV-rich(TIFF)Click here for additional data file.S2 FigPV mRNA, in budgerigar (parrot) pallial song nuclei . Sections modified from Chakraborty and colleagues (2015) with permission from Dr. Jarvis, who is also an author on the current paper. (B) Coronal sections of the woodpecker brain showing the analogous locations for dAN in the anterior nidopallium and dNA in the arcopallium. (C) Comparable coronal sections in a Harris hawk that show PV expression in many positive control areas (see (A) Representative radioactive in situ hybridization of microscope images of reas see . However(TIFF)Click here for additional data file.S3 FigPV mRNA expression (green) from fluorescent in situ hybridization experiments at low-magnification (tile scan) and (Ai and Ci) high-magnification illustrations of neuroanatomical regions with PV up-regulation in the male downy woodpecker brain. (E and F) Representative PV mRNA staining in the DLN, dAN, and dA of a female downy woodpecker. Blue signal is a DAPI nuclear stain. All scale bars are 1 mm. Asterisks (*) indicate folds on tissue. Photo credits: male downy woodpecker from Greg Schechter, and female downy woodpecker from Ken Thomas (CC Public Domain via WikiMedia).(A-D) Representative (TIF)Click here for additional data file.S4 FigPV or (C) arcopallium-enriched Lim homeobox 9 (Lhx9) mRNA expression. Medial parasagittal sections through the downy woodpecker arcopallium show that PV has specialized expression in dA of the dorsal and intermediate arcopallium. These findings are consistent with the 2 different types of probe labeling and hybridization methods. Although Lhx9 demarcates most of the woodpecker arcopallium, it is largely absent in the anterior arcopallium, as seen in songbirds [(A) Colormetric and (B) radioactive in situ hybridization illustrating specialized patterns of ongbirds . Dashed (TIFF)Click here for additional data file.S5 FigETV1 and (B) Lhx9 (radioactive in situ hybridization), were used to delineate the boundary of the arcopallium and nidopallium. (C-F) Representative in situ hybridization images (inverted black and white colormetric) of (C and D) FoxP1 and (E and F) RGS12 in zebra finch and downy woodpecker. Both genes are significantly enriched in the zebra finch Area X ; however, neither demarcates a specialized region within the woodpecker striatum. FoxP1 allows for the clear delineation of all nidopallial-striatal boundaries. Both reveal similar patterns to zebra finches hes (see ). Data f(TIF)Click here for additional data file.S6 FigEGR1 on adjacent parvalbumin (PV) sections in the dorsal arcopallial (dA) and drumming nucleus of the anterior nidopallium (dAN) or of male downy woodpeckers that were passively caught, low drummers and high drummers during simulated territorial intrusions (STIs). In situ hybridization microscope images of Arc on adjacent parvalbumin (PV) sections in the dorsal arcopallial (dA) and drumming nucleus of the anterior nidopallium (dAN) or of male downy woodpeckers that were passively caught, low drummers and high drummers during STIs. Violin plots of differences in EGR1 gene expression in the PV-rich (M) dA or (N) dAN nuclei, respectively, of male downy woodpeckers caught after producing different behaviors. EGR1 mRNA expression significantly differed in the dA , but we did not detect any differences EGR1 in the dNA across behavioral conditions . Data for M and N can be found in In situ hybridization microscope images of (TIF)Click here for additional data file.S7 Fig(A-B) Each recording includes an example of a stimulus drum (orange rectangles) being broadcast over a speaker and a resident (red rectangles) responding to this stimulus by producing a drum.(TIFF)Click here for additional data file.S8 FigData for these analyses can be found in (TIFF)Click here for additional data file.S1 TablePV expression present in a brain area is denoted by a plus (+) sign; PV absent in a brain area is denoted by a minus (\u2013) sign.(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S1 Data(CSV)Click here for additional data file.S2 Data(CSV)Click here for additional data file."} +{"text": "Until recently, the molecular aetiology of paediatric pulmonary hypertension (PH) was relatively poorly understood. While the TGF-\u03b2/BMP pathway was recognised as central to disease progression, genetic analyses in children were largely confined to targeted screening of risk genes in small cohorts, with clinical management extrapolated from adult data. In recent years, next-generation sequencing has highlighted notable differences in the genetic architecture underlying childhood-onset cases, with a higher genetic burden in children partly explained by comorbidities such as congenital heart disease. Here, we review recent genetic advances in paediatric PH and highlight important risk factors such as dysregulation of the transcription factors SOX17 and TBX4. Given the poorer prognosis in paediatric cases, molecular diagnosis offers a vital tool to enhance clinical care of children with PH. Current Opinion in Genetics & Development 2022, 75:101936Molecular and genetic basis of diseaseThis review comes from a themed issue on Neil Hanchard and Heather MeffordEdited by Molecular and Genetic Basis of Disease\u201dFor complete overview of the section, please refer to the article collection, \u201cAvailable online 27th June 2022https://doi.org/10.1016/j.gde.2022.101936http://creativecommons.org/licenses/by/4.0/).0959-437X/\u00a9 2022 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license is a rare, progressive and usually fatal disorder with an estimated annual incidence of 4\u201310 cases/million and a prevalence of 20\u201340 cases/million in children PAH has a complex aetiology with different factors precipitating disease presentation in adult- and childhood-onset disease, resulting in a varied pathobiology, disease severity and genetic background . The disIn this review, we summarise the key genes and pathways dysregulated in paediatric PAH with a focus on recent advances that highlight the applicability of molecular insights to clinical management.BMPR2 haploinsufficiency as a molecular mechanism of disease, additional causal variants were described in genes encoding the ALK1 (ACVRL1) and Endoglin (ENG) receptors, which have been observed in association with hereditary haemorrhagic telangiectasia (HHT). By contrast, initial reports of pathogenic variation in the type I receptor ALK6 (BMPR1B) have not been robustly validated, indicating a limited role for this gene in paediatric PAH The significance of canonical BMP/TGF-\u03b2 signalling in the pathogenesis of both adult and paediatric PAH has been well documented BMPR2 or ACVRL1, with or without HHT, have poor clinical outcomes by comparison to noncarriers, typically experiencing a marked limitation of physical activity within 3 years of diagnosis, highlighting the need for rapid mutation detection and treatment BMPR2 comprises structural variants that require additional analysis beyond targeted exonic sequencing Children with pathogenic variants in SMAD9) and caveolin-1 (CAV1), identified by candidate gene screening and whole-exome sequencing (WES), respectively. SMAD8 is a BMP-specific signalling intermediary whereas CAV1 plays an important role in regulation of the TGF-\u03b2 signalling system. Heterozygous variants in these genes have been reported to cause PAH in some paediatric-onset cases CAV1 may additionally be associated with a neonatal onset lipodystrophy syndrome Additional risk factors impacting BMP/TGF-\u03b2 signalling in PAH include SMAD8 (GDF2) gene are significantly enriched in adult-onset disease, based on exome-wide gene burden testing, and represent between 0.8% and 6.6% of all PAH cases with over 60 pathogenic variants reported to date BMP10 variation is also starting to emerge with four likely deleterious variants described, of which two were detected in childhood-onset PAH GDF2 mutation carriers, likely due to impaired cellular processing and secretion BMP9 and BMP10 are circulating ligands that specifically activate the ALK1/BMPR2 receptor complex. Rare heterozygous variants in the BMP9 ATP13A3 identified one homozygous and two compound heterozygous variants, of which p.Met850Ilefs*13 had been previously reported as heterozygous in an adult-onset case ATP13A3 variants cause a severe, autosomal recessive (AR) form of paediatric PAH KCNK3 was one of the first non-BMP pathway genes to be identified as a novel cause of autosomal dominant (AD) PAH by WES in a predominantly adult-onset I/HPAH cohort. The gene encodes a hypoxia-sensitive potassium channel involved in the regulation of the resting membrane potential of pulmonary arterial smooth muscle cells and pulmonary vascular tone Kcnk3 rat model, which demonstrates pulmonary vascular abnormalities consistent with PAH ABCC8 is a member of the ABC family which encodes sulfonylurea receptor 1 (SUR1) protein, a regulatory subunit of the KATP channel that controls channel function and potassium ion (K+) transport KCNK3 and ABCC8 in PAH provides novel exploratory avenues for molecular pathogenesis and may point to complementary or redundant functions of KCNK3 and KATPKCNA5 have also been implicated as potential modifiers in PAH pathogenesis, including in an early-onset case of severe PAH with a pathogenic BMPR2 variant The identification of loss-of-function variants of KDR which is characterised by PAH with low diffusing capacity for carbon monoxide (DLCO) KLK1) and gamma glutamyl carboxylase (GGCX) FBLN2) and platelet-derived growth factor D (PDGFD) de novo variants in additional genes, potentially explaining ~15% of paediatric PAH cases in their cohort. Given the limited time since initial discovery, further studies will be required to fully elucidate the role of these newly described genes in the pathogenesis of paediatric PAH.Recent novel gene discoveries include independent reports of heterozygous loss-of-function variants in kinase insert domain receptor, CPS1, NOTCH3, SMAD9 and the hypoxia-related genes TTLL3 and ITGAM, these remain to be independently validated A particular consideration in neonatal- and paediatric-onset pulmonary hypertension (PH) is the presence of complex comorbidities, often influenced by early developmental impacts on lung growth. The most common cause of transient PAH in neonates is persistent pulmonary hypertension of the newborn (PPHN), a failure of immediate postnatal cardiopulmonary transition with an annual incidence of 30.1 cases/million PH is also seen in children with connective tissue disease (APAH-CTD) or developmental lung diseases such as alveolar capillary dysplasia, bronchopulmonary dysplasia and congenital diaphragmatic hernia BMPR2, ENG, SMAD9, CAV1 and BMP10 have been linked to progressive PAH in children or young adults SOX17 was identified as a PAH risk gene by gene burden testing in an IPAH cohort SOX17 as a major cause of APAH-CHD, with the identification of rare deleterious missense variants in 9/13 paediatric cases SOX17 and TBX4, both inherited from an unaffected parent Childhood forms of PAH are frequently associated with congenital heart disease (CHD), which forms Group 1.4.4 of the sixth World Symposium on Pulmonary Hypertension classification . GermlinSOX17 encodes an endothelial transcription factor belonging to the SRY-related HMG box gene family involved in vascular development. SOX17 and its homologue SOX18 are crucial during embryonic development in processes such as angiogenesis, arterial specification and pulmonary vascular morphogenesis NOTCH1 and NOTCH3 have been described in childhood-onset PAH. NOTCH1 haploinsufficiency is a major cause of both nonsyndromic CHD and Adams\u2013Oliver syndrome; the identification of rare NOTCH1 variants in children with APAH-CHD would therefore be consistent with a secondary form of PAH, albeit in relatively few cases NOTCH3 variation remains limited TBX4 gene encodes a member of the T-box family of transcription factors that, together with TBX5, has an essential role in development of the limbs and respiratory system TBX4 variants cause ischiocoxopodopatellar syndrome , also known as small patella syndrome, whilst homozygous loss-of-function variants underlie posterior amelia with pelvic and pulmonary hypoplasia syndrome . Heterozygous TBX4-containing deletions or likely pathogenic TBX4 variants have been confirmed as a substantial cause of paediatric PAH, accounting for up to 8% of familial and idiopathic disease, with or without ICPPS TBX4 carriers is younger than BMPR2 carriers, demonstrating a significant enrichment in childhood- versus adult-onset cases TBX4 ranges from single nucleotide variants to large (>2\u2009Mb) deletions, encompassing multiple genes. The latter are more commonly associated with developmental delay and additional neurological and psychomotor defects, whereas PAH is typically caused by protein-truncating or deleterious missense variants TBX4 disruption represent a broad spectrum, ranging from transient neonatal PH to severe developmental lung disorders and progressive or biphasic PH, which may be associated with skeletal, cardiac and/or neurological anomalies. Moreover, TBX4 variants underlie a wide spectrum of clinicopathological outcomes. Neonatal respiratory failure has been reported due to lethal lung hypoplasia, such as acinar dysplasia or congenital alveolar dysplasia. By contrast, chronic PH can be diagnosed later in infancy and may recur in children previously recovered from PPHN TBX4 syndrome\u2019 characterised by severe pre-capillary PH points to a complex aetiology and disease progression that should be supported by regular cardiopulmonary assessment and multisystem imaging The EIF2AK4) gene have been described in children and young adults Pulmonary veno-occlusive disease and pulmonary capillary haemangiomatosis (PCH) are clinically similar subtypes of PAH with AR inheritance. Although histologically distinct, PCH and PVOD include venous and/or capillary abnormalities that are now clinically classified under PAH Group 1.6 EIF2AK4 variants are associated with low DLCO, which is a characteristic feature of PVOD/PCH EIF2AK4. However, it remains unclear whether these variants are a rare cause of AD PAH or potential genetic modifiers. Cases of clinically diagnosed PAH in whom biallelic EIF2AK4 variants have been detected are likely to represent previously misdiagnosed PVOD/PCH BMPR2 variation remains the predominant cause, the vast majority of newly reported genes each describe less than 3% of paediatric cases , the 10.13039/100004440Wellcome Trust, the Government Department of Business, Energy and Industrial Strategy (BEIS), the 10.13039/501100000274British Heart Foundation and 10.13039/501100000361Diabetes UK [SBF005\\1115]. For the purpose of Open Access, the author has applied a CC BY public copyright licence to any Author Accepted Manuscript (AAM) version arising from this submission.This work was supported by the Springboard Scheme Funders, namely the Fatima Taha: Investigation, Visualization, Writing \u2013 original draft; Laura Southgate: Conceptualization, Formal analysis, Visualization, Writing \u2013 original draft, Writing \u2013 review & editing.The authors declare no conflict of interest.Papers of particular interest, published within the period of review, have been highlighted as:\u2022 of special interest\u2022\u2022 of outstanding interest."} +{"text": "Home-delivered meals supported by the Older Americans Act (OAA) serve a dual purpose of improving nutritional intake and providing regular social contact for older adults. This regular contact can increase feelings of safety experienced by meal recipients. The benefits of home-delivered meal services may vary between meal recipients based on sociodemographic characteristics. Variation in home-delivered meal clients\u2019 reports of feeling safer at home because of regular meal delivery visits was examined to support ongoing efforts to increase social engagement and equity through the delivery of OAA services. Using data from the 2019 National Survey of Older Americans Act Participants (NSOAAP) home-delivered meal module, descriptive statistics and logistic regression were conducted to identify the characteristics associated with feeling safer at home because of meal delivery visits. The majority (85%) of meal recipients report feeling safer because of meal delivery visits, and rates were especially high for rural recipients (92%), those with a high school education or less (89%),racial/ethnic minorities (94%), and those with three or more ADL limitations (90%). Logistic regression found that rural residence (OR=3.3), lower educational attainment (OR=2.0), racial/ethnic minority status (OR=4.7), living alone (OR=1.6), and having 3+ ADLs (OR=1.9) were significantly associated with higher odds of feeling safer at home because of meal delivery visits; however, age, gender, and suburban residence were not significant. Findings suggest that benefits of home-delivered meal programs are supporting the needs of traditionally disadvantaged groups and broadly increasing meal recipients\u2019 sense of safety in their homes."} +{"text": "Staphylococcus aureus (MRSA) bacteremia. Animal models suggest ceftaroline penetrates the central nervous system (CNS) and vitreous humor; however, there are limited data which describe outcomes of patients treated with ceftaroline for infections with CNS or ocular involvement.Ceftaroline is increasingly used in combination therapy for methicillin resistant > 18 years who received > 48 hours of ceftaroline and had evidence of CNS or ocular involvement were included. Descriptive statistics were used.This was a single-center, retrospective study at an academic medical center. The primary objective was to describe outcomes in patients treated with ceftaroline for CNS or ocular infections. Patients who received ceftaroline from January 2015 to February 2022 were identified. Patients Staphylococcus epidermidis (Table\u00a02). Ceftaroline was salvage therapy in nine patients (90%), often following vancomycin and used as part of combination therapy (Table\u00a03).Ten patients met the inclusion criteria: six with CNS and four with ocular infections. The cohort was predominantly white (90%) and male (70%). The most common comorbidities were hypertension (70%) and diabetes (40%) (Table\u00a01). Nine patients were treated for MRSA and one patient was treated for methicillin resistant Of the four patients with ocular infections, all had MRSA bacteremia. Two patients (50%) had positive vitreal cultures and received intra-vitreal injections (Table\u00a02). Most of these patients (75%) died during admission (Table\u00a04). Of the six patients with CNS infections, injection drug use was more common than in patients with ocular involvement (33% vs 0%), as was immunocompromise (50% vs 0%) and the presence of endocarditis (33% vs 0%). Of the patients with CNS involvement, three had abnormal lumbar punctures and the remainder were diagnosed by CNS imaging (Table\u00a02). Four patients with CNS involvement survived hospital stay (67%), and one of these had recurrence of CNS infection within 90 days (Table\u00a04).Although patients with CNS or ocular involvement represent a severely ill subset of staphylococcal infections, ceftaroline is a promising agent in combination therapy. Comparative data is needed to validate these findings.All Authors: No reported disclosures."} +{"text": "Scientific Reports 10.1038/s41598-022-22239-4, published online 15 October 2022Correction to: The Competing interests section in the original version of this Article was incorrect.\u201cThe authors declare no competing interests.\u201dnow reads:\u201cThe authors declare no competing interests. The views expressed in this article are the authors only and do not reflect the views of the European Food Safety Authority.\u201dThe original Article has been corrected."} +{"text": "OOpioid system activity was found disturbed in several reward circuit areas in restrictive anorexia nervosa (AN) patients but also at the pituitary level. The role of this specific abnormality in AN physiopathology remains unknown.We aimed to evaluate the relationship of upper mentioned AN abnormality with its classical pituitary features and eating behavior traits.11C] diprenorphin binding potential (BPND) were processed for each pituitary part in three groups of young women: 12 AN, 11 recovered AN patients (ANrec), and 12 Controls. Anterior pituitary hormones and neurohypophysis (NH) 12 points circadian profile including copeptin and oxytocin, psychological scores were evaluated in these subjects as well as in 13 bulimic (BN) patients.PET [11C] diprenorphin pituitary binding was found to be fully localized in NH. Only AN patients\u2019 NH present lower [11C] diprenorphin BPND than Controls, interpreted as a higher opioid tone. Both AN and ANrec show lower copeptin/24h than in Controls but no difference in oxytocin. BN showed increased copeptin and low oxytocin. In AN patients copeptin inversely correlate with Restrained Eating while oxytocin correlate with the External Eating score. NH [11C] diprenorphin BPND correlated with leptin but not with copeptin or oxytocin.[Neurohypopysis opioid tone in anorexia nervosa seem not to impact the vasopressin or oxytocin release but still may interfere in gonadal axis regulation. Copeptin, a good indicator of hydration state, may be a good tool to detect hidden restrictive or purging behaviors. Specific correlates with AN psychologic features still suggest a physiopathological involvement.No significant relationships."} +{"text": "Using population-wide administrative data, the objective was to provide Canadian evidence on the longitudinal relationship between maternal intimate partner violence (IPV) victimization and children\u2019s developmental health. Using provincial prosecution records, we examined developmental vulnerability (DV) at kindergarten of children prenatally exposed to maternal IPV victimization compared to unexposed counterparts.This retrospective cohort study linked administrative datasets from the Population Research Data Repository at the Manitoba Centre for Health Policy. Exposed mother-child pairs with 1+ prosecution record of maternal IPV victimization during pregnancy between 2003 and 2018 in Manitoba were matched to unexposed pairs (1:3) based on sex/birthdate of child and neighbourhood income. DV at kindergarten was measured across 5 domains using the Early Developmental Instrument (EDI). Children without eligible EDI scores were excluded. Multiple logistic regression models were conducted to address the objective.The eligible cohort included 927 children ; 31.07% of the cohort was developmentally vulnerable in one or more domains (1/+) and 19.53% was developmentally vulnerable in two or more domains (2/+). Children who were prenatally exposed to maternal IPV victimization had increased odds of vulnerability across all 5 developmental domains . Unadjusted ORs showed statistically significant associations between maternal exposure of prenatal IPV victimization and DV in 1/+ and 2/+ . When adjusted for covariates , no statistically significant relationship was found for any of the domains , 1/+ , and 2/+ .The unadjusted, statistically significant associations suggest children exposed to maternal IPV victimization prenatally may face associated social/health risks. The finding highlights the need to consider potential factors that put children at risk of DV when developing and implementing support systems/interventions for children exposed to maternal IPV victimization."} +{"text": "Cortisol, the hormonal endpoint of Hypothalamic Pituitary Adrenal (HPA) axis, coordinates the body response in front of daily stressful situations. Disturbances in cortisol circadian rhythm have been implicated in the pathophysiology of depression and neurodevelopment lasting consequences. Although pregnancy entails a progressively increase in cortisol levels, the consequences of subclinical depression traits during pregnancy in cortisol circadian rhythm remains unclear.To analyze the impact of prenatal subclinical depressive symptomatology in cortisol circadian rhythm through pregnancy and its relevance for postpartum depression risk.A cohort of 112 healthy pregnant women (Mean age\u00b1SD=32.32\u00b14.37) of the general population was followed throughout their first pregnancy and first two months of postpartum period. Diurnal salivary cortisol curve (four measures) was obtained for every trimester; the Area Under the Curve with respect to the ground (AUCg) and with respect to the increase (AUCi) were used as measures of basal HPA axis functioning. Depressive symptomatology was assessed every pregnancy trimester and postpartum period following EPDS criteria. All the analyses were adjusted for maternal age, weight, ethnicity and socioeconomic status and sample collection\u2019s time.2=13.8, p<.001,OR=9.6; 95%CI 2.5\u201335.5).Prenatal subclinical depressive symptomatology (EPDS>10) was associated with a blunted cortisol rhythm during first trimester but not during second or third trimesters . Furthermore, a logistic regression model showed a positive association between Prenatal subclinical depressive symptomatology and the risk of postpartum depression (\u03c7Women with subclinical depressive symptomatology in early pregnancy had alterations in cortisol circadian rhythmicity and a higher risk of postpartum depression.No significant relationships."} +{"text": "To what extent are shared genetic determinants in the comorbidities and associations between gastrointestinal tract diseases and psychiatry disorders involved in the gut-brain axis?In this genome-wide pleiotropic association study using genome-wide association summary statistics from publicly available data sources, pervasive genetic correlations and genetic overlaps between gastrointestinal tract diseases and psychiatric disorders were found. The pleiotropic genetic determinants between them were extensively distributed across the genome.These findings not only support the shared genetic basis underlying the gut-brain axis but also have important implications for intervention and treatment targets of these 2 types of diseases simultaneously. This genome-wide pleiotropic association study sequentially investigates the pleiotropic associations from genetic and biological pathways to disentangle the underlying shared genetic etiology between 4 gastrointestinal tract diseases and 6 psychiatric disorders. Comorbidities and genetic correlations between gastrointestinal tract diseases and psychiatric disorders have been widely reported, with the gut-brain axis (GBA) hypothesized as a potential biological basis. However, the degree to which the shared genetic determinants are involved in these associations underlying the GBA is unclear.To investigate the shared genetic etiology between gastrointestinal tract diseases and psychiatric disorders and to identify shared genomic loci, genes, and pathways.This genome-wide pleiotropic association study using genome-wide association summary statistics from publicly available data sources was performed with various statistical genetic approaches to sequentially investigate the pleiotropic associations from genome-wide single-nucleotide variation , and gene levels and biological pathways to disentangle the underlying shared genetic etiology between 4 gastrointestinal tract diseases and 6 psychiatric disorders . Data were collected from March 10, 2021, to August 25, 2021, and analysis was performed from January 8 through May 30, 2022.The primary outcomes consisted of a list of genetic loci, genes, and pathways shared between gastrointestinal tract diseases and psychiatric disorders.INAVA), 19q13.33 (FUT2), 11q23.2 (NCAM1), and 1p32.3 (LRP8).Extensive genetic correlations and genetic overlaps were found among 22 of 24 trait pairs. Pleiotropic analysis under a composite null hypothesis identified 2910 significant potential pleiotropic SNVs in 19 trait pairs, with 83 pleiotropic loci and 24 colocalized loci detected. Gene-based analysis found 158 unique candidate pleiotropic genes, which were highly enriched in certain GBA-related phenotypes and tissues, whereas pathway enrichment analysis further highlighted biological pathways primarily involving cell adhesion, synaptic structure and function, and immune cell differentiation. Several identified pleiotropic loci also shared causal variants with gut microbiomes. Mendelian randomization analysis further illustrated vertical pleiotropy across 8 pairwise traits. Notably, many pleiotropic loci were identified for multiple pairwise traits, such as 1q32.1 (These findings suggest that the pleiotropic genetic determinants between gastrointestinal tract diseases and psychiatric disorders are extensively distributed across the genome. These findings not only support the shared genetic basis underlying the GBA but also have important implications for intervention and treatment targets of these diseases simultaneously. The GBA is characterized by bidirectional interactions between the gastrointestinal tract and the central nervous system (CNS) and would link intestinal dysfunction and inflammation with brain function and psychiatric disorders.5 Various biological mechanisms were involved in the GBA, such as inflammatory immune responses, the autonomic nervous system, and enteric nervous system, where the role of the composition of gut microbiota and related metabolites has been particularly highlighted.7 Underlying the conceptual framework of the GBA, the shared genetic etiology might be involved in the associations between gastrointestinal tract diseases and psychiatric disorders.The comorbidities and associations between gastrointestinal tract diseases and psychiatric disorders have been widely reported,10 Genetic correlations have been suggested between these 2 types of diseases using linkage disequilibrium (LD) score regression (LDSC).14 However, it remains unclear whether the overall genetic correlation would be attributed to a few loci or across the genome.15 Indeed, there would be genetic overlap even without any genetic correlation. Although previous studies have investigated genetic overlap,16 shared susceptibility genes,18 and causal relationships21 between these 2 types of diseases, they mainly focused on inflammatory bowel disease (IBD) and psychiatric disorders with limited sample sizes. Recently, 2 studies22 have conducted GWAS of specific gastrointestinal tract diseases as well as systematic post-GWAS analyses and pointed out the necessity to explore the shared genetic risk across traits to improve the understanding of the disordered brain-gut interactions. Therefore, it is of great importance to further seek out the specific genomic variants or loci accounting for genome-wide genetic correlation and to deeply probe into the shared genetic etiology between these 2 types of diseases. Shared genetic etiology also indicates the potential pleiotropy, which often acts as genetic confounding of the associations between trait pairs.25 Cross-trait analysis has been proposed to investigate the pleiotropic genetic variants or loci among multiple traits by leveraging the correlation of GWAS signals,29 where the pleiotropic loci could serve as intervention targets with the potential to simultaneously prevent or treat these diseases.Genome-wide association studies (GWAS) have identified multiple genetic variants associated with gastrointestinal tract diseases and psychiatric disorders.In this genome-wide pleiotropic association study, using large-scale GWAS summary data, we performed a genome-wide pairwise trait pleiotropic analysis between 4 gastrointestinal tract diseases and 6 psychiatric disorders through various statistical genetic approaches to sequentially investigate the pleiotropic associations from genome-wide, SNV, and gene levels and biological pathways to disentangle the underlying shared genetic etiology. Of note, under the framework of pleiotropic analysis, we first performed the SNV-level analysis to detect pleiotropic variants and loci, followed by pairwise colocalization analysis to determine colocalized loci and gene-level analysis to identify candidate pleiotropic genes, based on which we further performed parallel phenotype and tissue-specific enrichment analysis to characterize the phenotype and tissue specificity as well as additional gene-level analysis to identify the tissue-specific and cell type-specific pleiotropic genes. We also highlighted the role of gut microbiomes in interpreting the shared genetic etiology, followed by mendelian randomization analysis to evaluate pairwise causal associations and partly characterize different types of pleiotropy .10 GWAS for IBS were obtained from a larger meta-analysis with 486 601 individuals (53 400 cases and 433 201 controls).22 GWAS for the 6 psychiatric disorders were from the Psychiatric Genomics Consortium, including schizophrenia,30 BIP,31 MDD,32 ADHD,33 PTSD,34 and AN.35 In addition, GWAS for early age-related macular degeneration (AMD)36 and cataract37 were obtained to serve as a common set of negative controls for both gastrointestinal tract diseases and psychiatric disorders, given these 2 disorders are relatively limited to the pathological lesions of intraocular contents, and previous studies also showed no significant genetic correlations between AMD and MDD38 as well as among cataract, GERD, and psychiatric symptoms.39 All GWAS were approved by relevant ethic committees, and written informed consent was obtained from all participants, with details provided in the eTable 1 in STREGA) reporting guideline.We sought GWAS summary statistics from publicly available data sources with European ancestry, owing to the limited availability of well-powered GWAS with other ancestries, and selected GWAS with sample sizes larger than 50 000 to ensure statistical power. GWAS for GERD, IBD, and PUD were obtained from the same gastrointestinal tract GWAS based on 456 327 individuals from UK Biobank (UKB).All analyses were performed after excluding SNVs in the major histocompatibility complex region (chromosome 6: 25-35\u2009megabase [Mb]) due to its complex LD structure and restricted to biallelic SNVs with minor allele frequency larger than 0.01. Details of these methods are provided in 40 and high-definition likelihood41 were used to assess genome-wide genetic correlations for 24 pairwise traits. The intercept from LDSC could also indicate potential sample overlap between 2 GWAS. Given that genetic correlation only reflects the overall correlation across the genome, we further applied genetic analysis incorporating pleiotropy and annotation (GPA)29 to explore the overall genetic overlap between traits. The Bonferroni-corrected significant threshold was set at P\u2009<\u20092.08\u2009\u00d7\u200910\u22123 (.05/24). In addition, negative control analysis was performed through LDSC between AMD and cataract and the 4 gastrointestinal tract diseases as well as the 6 psychiatric disorders, with Bonferroni-corrected significant threshold set at P\u2009<\u20092.5\u2009\u00d7\u200910\u22123 (.05/20).Both LDSC26 Single-nucleotide variations with P\u2009<\u20095\u2009\u00d7\u200910\u22128 for PLACO were considered significant pleiotropic variants. Functional mapping and annotation of genetic associations (FUMA)42 was applied to characterize potential pleiotropic loci, based on which a bayesian colocalization analysis43 was performed to further identify shared causal variants in each pleiotropic locus. We declared a colocalized locus with posterior probability of H4 (PP.H4) larger than 0.7.For the union set of pairwise traits with significant genetic correlation or genetic overlap, we used pleiotropic analysis under composite null hypothesis (PLACO) to identify potential pleiotropic SNVs.44 analysis on the genes located in or overlapped with the pleiotropic loci based on both PLACO outputs and single-trait GWAS to identify candidate pleiotropic genes, with the significance declared at the locus-specific Bonferroni-corrected 2-sided P\u2009<\u2009.05 for MAGMA analysis on PLACO results and 2-sided P\u2009<\u2009.05 for MAGMA analyses based on original single-trait GWAS. Further, we performed phenotype enrichment analysis based on the Mouse Genome Informatics platform45 to characterize the phenotype specificity of these pleiotropic genes against that of nonpleotropic genes by examining the differences in the proportions of genes associated with certain phenotypes in the pleiotropic gene group against that in the nonpleotropic gene group using the Fisher exact test. Then, we performed tissue-specific enrichment analyses to illustrate the tissue specificity of these pleiotropic genes using the deTS tissue-specific enrichment method46 based on 2 different reference panels, Genotype-Tissue Expression project (GTEx)47 and the Encyclopedia of DNA Elements project (ENCODE).48 We declared the significance with a nominal threshold (2-sided P\u2009<\u2009.05) for these 2 parallel enrichment analyses. In addition, we used E-MAGMA (expression quantitative trait loci [eQTL]\u2013informed MAGMA)49 and transcriptome-wide association study analysis using joint-tissue imputation50 to further investigate the tissue-specific genes, parallelized with H-MAGMA (Hi-C\u2013coupled MAGMA)51 to indicate the cell-type specificity, in which we declared the significance with locus-specific Bonferroni correction. Gene set enrichment analysis was also performed to identify potential biological pathways using the clusterProfiler package.52 Significantly enriched pathways were declared with normalized enrichment score greater than 2 and adjusted P\u2009<\u2009.05. Multitrait colocalization analysis with HyPrColoc 53 incorporating host-microbiome GWAS was performed to highlight the critical role of gut microbiome.Based on PLACO results, we further explored the shared biological mechanisms of these pleiotropic loci. We performed gene-level multimarker analysis of GenoMic annotation (MAGMA)54 as main analysis and in total 6 alternative mendelian randomization methods with different model assumptions as additional analyses for further validation. For main analysis, we chose a false discovery rate approach for multiple testing correction with the significance threshold being false discovery rate\u2013adjusted P\u2009<\u2009.05, given that the commonly used Bonferroni correction is often too stringent for multiple nonindependent tests. The nominally significant threshold (P\u2009<\u2009.05) was used for alternative mendelian randomization methods. The Latent Heritable Confounder Mendelian Randomization (LHC-MR) method, which could account for sample overlap, was used for pairwise traits with potential sample overlap to further validate the mendelian randomization results.55Last, we conducted bidirectional mendelian randomization analysis for 24 pairwise traits between gastrointestinal tract diseases and psychiatric disorders to investigate the potential causal trait pairs , along with negative control analysis using AMD and cataract. We used the inverse-variance weighted method42 We characterized independent SNVs with r2 less than 0.6000 and lead SNVs with r2 less than 0.1000 within 1 Mb. Genomic risk loci were defined by merging genomic regions if the physical distance between lead SNVs was less than 250 kilobase.42For significant pleiotropic SNVs from PLACO, we applied FUMA to identify independent variants, characterize genomic risk loci, and annotate the functions of variants using LD information from the 1000 Genome Project phase 3 reference panel of European population.42 Single-nucleotide variations with P\u2009<\u20095\u2009\u00d7\u200910\u22128 in each single-trait GWAS were also annotated by FUMA for comparison.Functional annotations, including ANNOVAR software tool categories (Bioinformatics), combined annotation-dependent depletion (CADD) scores, and RegulomeDB scores, were also provided by FUMA. Single-nucleotide variations with a CADD score larger than 12.37 were considered a potentially deleterious variant.Using genome-wide association summary statistics from publicly available data sources, pervasive significant genome-wide genetic correlations and genetic overlaps were found across 24 pairwise traits, among which 14 were identified from LDSC and 21 were identified from GPA . NotablyNCAM1 [OMIM 116930]) and 1q32.1 (mapped gene: INAVA [OMIM 618051]), suggesting the extensive pleiotropic effects of these loci. The top SNVs in 83 FUMA-annotated pleiotropic loci showed mixed directions of allelic associations were identified as potential pleiotropic variants by PLACO in 19 trait pairs was a significant eQTL for EP300 (OMIM 602700) encoding p300 protein, which plays an important role in cell proliferation and differentiation. The index SNVs rs681343 (PLACO P\u2009=\u20091.36\u2009\u00d7\u200910\u221212 for PUD-schizophrenia) and rs601338 (PLACO P\u2009=\u20095.07\u2009\u00d7\u200910\u221211 for PUD-ADHD) at 22q13.2 were in high LD (r2\u2009=\u20090.996) and had almost identical eQTL regulation information (eTable 7 in FUT2 (OMIM 182100). The index SNV rs13107325 at 4q24 locus was a significant eQTL for SLC39A8 (OMIM 608732), which encodes ZIP8 metal cation transporter. In addition, 7 index SNVs (6 unique) were identified with CADD scores larger than 12.37, in which 2 mRNA exonic variants had higher CADD scores, including rs601338 and rs13107325 .ANNOVAR category annotation illustrated that 34 of 83 index SNVs (41%) were intronic variants and 27 of 83 (33%) were intergenic variants. Only 7 of 83 index SNVs (8%) (6 unique) were exonic variants, including 5 messenger RNA (mRNA) exonic variants and 2 noncoding RNA exonic variants (eTable 5 in FUT2). In addition, 7 pleiotropic loci were identified with PP.H3 larger than 0.7000, indicating there might be different causal variants in these loci with PP.H4 larger than 0.7, in which 22 top SNVs of corresponding loci were identified as candidate shared causal variants . InteresNCAM1 was identified as a significant pleiotropic gene in 5 pairs of traits, followed by INAVA, CACNA1S (OMIM 114208), and KIF21B (OMIM 608322) in 4 pairs of traits and BANK1 (OMIM 610292), CAMKV (OMIM 614993), DPYD (OMIM 612779), MON1A (OMIM 611464), MST1R (OMIM 600168), PSCA (OMIM 602470), RBM5 (OMIM 606884), RBM6 (OMIM 606886), and SLC39A8 in 3 pairs of traits.MAGMA analysis based on 295 potential pleiotropic genes that located in or overlapped with 83 pleiotropic loci identified 196 significant pleiotropic genes (158 unique), in which 38 genes were detected in 2 or more trait pairs (eTable 9 in P\u2009=\u20091.80\u2009\u00d7\u200910\u22122) B. Detail\u2009\u00d7\u200910\u22122) in sever\u2009\u00d7\u200910\u22122) .PSCA (8q24.3) for 3 pairwise traits , LPR8 (1p32.3) for 2 pairwise traits (IBS-schizophrenia and IBS-BIP), and FUT2 (19q13.33) for 2 pairwise traits (PUD-schizophrenia and PUD-ADHD). H-MAGMA analysis further suggested the cell-type specificity of these pleiotropic genes. Details of these gene-level analyses were also provided across 8 pleiotropic loci were significantly detected in multiple GBA-related tissues in at least 2 trait pairs, for example, P\u2009<\u2009.05), including 99 Gene Ontology (GO) terms and 28 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways (eTable 16 in H1) and TH2 cell differentiation (KEGG has04658) and TH17 cell differentiation (KEGG has04659), which involve in immunoinflammatory responses, were highlighted in 2 pairs of traits.Gene set enrichment analysis identified 127 significantly enriched pathways was also identified as shared causal variant.Multitrait colocalization analysis from HyPrColoc highlighted 15 pleiotropic loci (11 unique) that were colocalized to share a causal variant, supporting the significant role of gut microorganisms . Seven oBidirectional mendelian randomization analyses using inverse-variance weighted method showed 11 significant positive associations , 19q13.33 (FUT2), 11q23.2 (NCAM1), and 1p32.3 (LRP8). Several loci previously identified to be associated with gastrointestinal tract diseases were illustrated to be potential pleiotropic loci shared with psychiatric disorders and vice versa (eDiscussion in Overall, pleiotropic variants between gastrointestinal tract diseases and psychiatric disorders were extensively distributed, with several loci especially highlighted between certain trait pairs, such as 1q32.1 (H1, TH2, and TH17 cell differentiation pathways were highlighted between IBD and schizophrenia, BIP, and AN. The TH17 cells play important roles in the development of inflammatory responses and autoimmune diseases.58 Previous studies suggested the significant role of dysfunction of TH17 cell differentiation and the accumulation of TH17-related cytokines in the pathological process of IBD.60 In addition, TH17 cells and related cytokines would provoke CNS neuroinflammation and neurotoxicity under pathological status and disrupt the blood-brain barrier and thus alter the permeability of the blood-brain barrier.62 Bacterially produced bile acid metabolites have been shown to inhibit TH17 cell differentiation, which may be related to the pathophysiology of IBD.63 Therefore, TH17 cell differentiation and function play a vital role in the regulation of brain-gut-microbiome axis.Shared genetic determinants also reflect common biological pathways, among which the TOur study is not without limitations. First, we were unable to assess the causal effects of gut microbiome on gastrointestinal tract diseases or psychiatric disorders through mendelian randomization analyses since SNVs with associated gut microorganisms were hard to obtain due to the limited sample size of gut microbiome GWAS. Therefore, caution should be used on the interpretation of the role of gut microorganisms. Second, our study was restricted to European ancestry and may not generalize to other ancestries.Given strong evidence of genetic correlation and genetic overlap between gastrointestinal tract diseases and psychiatric disorders, we found that pleiotropic genetic variants, loci, and genes were extensively distributed across the genome, with higher phenotype and tissue specificity. We highlighted some genetic determinants previously associated with gastrointestinal tract diseases that were shared with psychiatric disorders and vice versa. More importantly, shared biological mechanisms concerning immune response, synaptic structure and function, and potential gut microbiome were identified. Our findings not only support the shared genetic basis underlying GBA but provide novel insight into the intervention and treatment targets of these diseases."} +{"text": "This correlationwas significantly reduced when using acomputational method for suppression of macromolecular signals knownas small molecule enhancement spectroscopy (SMolESY) for proteinicbaseline removal prior to PQN. These results highlight proteinuriaas a common yet overlooked source of bias in 1H NMR metabolicprofiling studies which can be effectively mitigated using SMolESYor other macromolecular signal suppression methods before estimationof normalization coefficients.Normalizationto account for variation in urinary dilution is crucialfor interpretation of urine metabolic profiles. Probabilistic quotientnormalization (PQN) is used routinely in metabolomics but is sensitiveto systematic variation shared across a large proportion of the spectralprofile (>50%). Where Probabilistic quotientnormalization (PQN)11 leverages the datacaptured in the metabolic profile as a whole and is the \u201cgoldstandard\u201d for computational normalization of 1HNMR spectra in urinalysis studies. PQN uses the complete set of profilemeasurements to estimate a normalization coefficient for each samplethat is representative of its dilution factor relative to a predefinedreference . Compared to other normalizationmethods , PQN provides robustnessagainst bias from few signals whose intensity dominates the totalprofile integral. However, PQN is not suitable if a large proportionof variables (50% or more) covary systematically with factors otherthan the sample dilution.11Urine exhibits strong intradayand interindividual variation indilution owing to factors such as hydration status, kidney function,diet, medication, and voiding interval. This poses a fundamental challengein urinalysis, especially of spot urine samples, that must be accountedfor when accurately making or comparing chemical measurements1H NMR-basedmetabolic profiles by elevating the spectral baselineand contributing broad signals in a concentration-dependent manner admitted to hospital with COVID-19,collected by the International Severe Acute Respiratory and EmergingInfections Consortium (ISARIC) following the WHO Clinical CharacterizationProtocol UK (CCP-UK). Further NMR experimental details are reportedin the Supporting Information. 1H NMR profiles (n = 85) from COVID-19 patients,focusing on the backbone \u2212NH and parts of methyl proteinicprotons, clearly illustrating the effects of proteinuria on the smallmolecule profiles observed. The impact of macromolecular signals onthe small molecule profile may be reduced when employing additionalNMR experiments beyond the routine one-dimensional general profileexperiment spectral editing experiments such as spin\u2013echo pulse sequences19). However, the need for these additional experimentsmay not be anticipated at the outset of a urine profiling study, andtheir use both increases the experimental cost and decreases analysisthroughput. We recently introduced small molecule enhancement spectroscopy(SMolESY)12 as a computational alternativefor removal of macromolecule-derived signals directly from routineone-dimensional NMR spectral profiles without the need of extra experiments,enabling the more specific and direct analysis of small molecule analytes. The approach can also be effectivelyreversed, removing the sharp small molecule-derived signals and providingan enhanced protein baseline for the purposes of urinary protein quantification(S3).20 Integrationof the proteinic methyl group-containing spectral region between 0.2and 0.5 ppm (1H NMR spectra and their SMolESY processed counterparts (r2 = 0.423) of the variance inPQN coefficients estimated from the 1D 1H NMR spectra withoutmacromolecular baseline removal (r2 = 0.163) when the macromolecular signatureis removed via SMolESY prior to PQN .We used an fication2a, S3.20 0.5 ppm 2a accura 0.5 ppm 2b. In thterparts 3. Althou removal 4a. This r to PQN 4b, closer2 = 0.43, r2 = 0.1, A comparison between the PQN coefficients and creatinineconcentrationsis shown in 21 or with computational methods.23 However, we advocate the use of SMolESY, because of its ease ofapplication to 1H NMR spectra (including retrospectiveapplication where proteinuria is observed after the fact), being fastand highly effective method for removing broad baseline signals fromprotein and improving the estimation of normalization coefficients.Our observations and proposed methodology are of high importance forthe accurate normalization of urine biofluid 1H NMR spectra,especially in the context of studies on diseases and phenotypes whereproteinuria is likely to be present .24Normalization procedures are crucial to correctlyinterpret urinarymetabolic profiles. Here, we show that protein signals can confoundprobabilistic quotient normalization, and it is reasonable to assumethis could also happen with other computational normalization methods.We recommend the removal of protein baseline signals prior to estimationof normalization coefficients. This can be performed experimentally"} +{"text": "Background:Staphylococcus aureus is the second-leading cause of late-onset sepsis among infants in US neonatal intensive care units (NICUs). Management of S. aureus bacteremia and meningitis in infants varies widely due to the lack of standardized guidelines. We examined the association between initial antibiotic duration and recurrent S. aureus infection or death among NICU infants with S. aureus bacteremia and/or meningitis. Methods: We conducted a retrospective cohort study of infants in Pediatrix Medical Group NICUs from 1997 to 2018 with first episode of S. aureus bacteremia and/or meningitis, identified as having at least 1 blood or cerebrospinal fluid (CSF) culture growing only S. aureus at any point during their NICU stay. Excluded infants were those not started on antistaphylococcal therapy within 2 days of positive culture, those with had endocarditis or osteomyelitis, or those who died or were discharged during or up to 1 day after antibiotic cessation. Antibiotic cessation was defined as last day of antibiotic given if followed by at least 3 days without antibiotics. Multivariable logistic regression was used to analyze the association between antibiotic duration categorized as <14 or \u226514 days) and recurrent SA infection , or death (within 7 days of antibiotic cessation and at discharge). Results: Of 4,086 infants included, 3,991 (98%) had S. aureus bacteremia only and 95 (2%) had meningitis \u00b1 bacteremia. Of those with bacteremia only, 2,017 (50.5%), and 17 (18%) of those with meningitis received <14 days antibiotics (Figure Conclusions: In the largest study thus far examining antibiotic duration among hospitalized infants with S. aureus bacteremia and/or meningitis, \u226514 days antibiotics was associated with decreased odds of recurrent infection or death. Further studies are needed to define the optimal treatment duration and identify clinical factors distinguishing infants able to safely receive a shorter antibiotic duration.Funding: NoDisclosures: None"} +{"text": "ABCB11 gene cause a spectrum of rare liver diseases. The most severe form is progressive familial intrahepatic cholestasis type 2 (PFIC2). Current medical treatments have limited efficacy. Here, we report the in vitro study of Abcb11 missense variants identified in PFIC2 patients and their functional rescue using cystic fibrosis transmembrane conductance regulator potentiators. Three ABCB11 disease-causing variations identified in PFIC2 patients were reproduced in a plasmid encoding an Abcb11-green fluorescent protein. After transfection, the expression and localization of the variants were studied in HepG2 cells. Taurocholate transport activity and the effect of potentiators were studied in Madin\u2013Darby canine kidney (MDCK) clones coexpressing Abcb11 and the sodium taurocholate cotransporting polypeptide (Ntcp/Slc10A1). As predicted using three-dimensional structure analysis, the three variants were expressed at the canalicular membrane but showed a defective function. Ivacaftor, GLP1837, SBC040 and SBC219 potentiators increased the bile acid transport of A257V and T463I and to a lesser extent, of G562D Abcb11 missense variants. In addition, a synergic effect was observed when ivacaftor was combined with SBC040 or SBC219. Such potentiators could represent new pharmacological approaches for improving the condition of patients with ABCB11 deficiency due to missense variations affecting the function of the transporter.ABCB11 is responsible for biliary bile acid secretion at the canalicular membrane of hepatocytes. Variations in the ABCB11, or the bile salt export pump (BSEP), is an adenosine tri-phosphate (ATP)-binding cassette (ABC) transporter localized at the apical plasma membrane of hepatocytes. It allows normal bile flow through its essential function of biliary bile acid (BA) secretion ,3. HowevABCB11 variations early during childhood TC (Perkin Elmer) was added in the basal compartment (in the wells). After two hours, the apical buffer (in the membrane inserts) was collected, and transcellular transport of [3H]TC was calculated from the radioactivity present in the apical buffer. Transport data were normalized to protein amounts determined by bicinchoninic acid assay .Measurement of Abcb11-GFP-mediated TC secretion was performed in polarized MDCK cells grown on membrane inserts, as described . In briep value < 0.05 was considered significant.Data are expressed as means \u00b1 standard error of the mean (SEM). Graphics and statistical analyses were performed using one-way ANOVA using Prism version 7.00 . A"} +{"text": "Pseudomonas sp. strains MWU12-2020 and MWU12-3103b, isolated from the rhizospheres of wild and cultivated cranberry bogs in southeastern Massachusetts; these strains are unrelated to known Pseudomonas species. The genomes of both isolates exceed 6\u2009Mbp and contain predicted ice nucleation and type VI and III secretion system genes.Here, we present the draft genome sequences of Pseudomonas allow them to interact and survive within many different environments and strain MWU12-3103b was isolated from cultivated cranberry bog roots during culture-dependent microbe surveys. Cranberry plants have a network of fine roots supported by ericoid mycorrhizae, which makes it difficult to differentiate between root and fungus and determine where the root zone is were used to extract genomic DNA (gDNA) from overnight KMB broth cultures. Libraries for genomic sequencing were generated from enzymatically sheared DNA (\u2248500\u2009bp) using a HyperPlus library preparation kit . The sheared fragments were end repaired and A-tailed at the 3\u2032 end. Indexed Illumina-compatible adapters (IDT number 00989130v2) were ligated to the A-tailed fragments; the library was cleaned using KAPA pure beads (KK8002) and then amplified using HiFi enzyme (KAPA KK2502). Fragment sizes were determined using an Agilent TapeStation device and then quantified using a KAPA quantitative PCR (qPCR) library quantification kit (KK4835) on a QuantStudio 5 system for sequencing on the Illumina MiSeq platform (2\u2009\u00d7\u2009250-bp format). Using the Comprehensive Genome Analysis feature of PATRIC v3.6.12 (https://www.patricbrc.org) (Pseudomonas by genome BLAST distance phylogeny using TYGS v342 (Pseudomonas koreensis DSM 16610T (GenBank accession number JAAQYM000000000) , neither can be assigned to an existing species, and they do not belong in the same species (dDDHd4\u2009=\u200951.2). Both genomes contain putative inaA genes for ice nucleation , 12 for cleation and typeation 16\u2013, all of JALMEX010000000 and JALMEY010000000 and BioProject accession number PRJNA691338 (https://zenodo.org/record/6412376#.Yxj1mEfMKUk).This whole-genome shotgun sequencing project has been deposited at DDBJ/EMBL/GenBank under accession numbers NA691338 . The ver"} +{"text": "Clostridioides difficile infection (CDI) are often limited in their ability to identify potentially important exposures occurring long before diagnosis. We describe hospitalizations and antibiotic use (AU) occurring up to one year prior to CDI diagnosis among Medicare beneficiaries.Studies describing risk factors for C. difficile test in a person \u226565 years without a positive test in the prior 8 weeks) identified during 2016\u20132018 through population-based CDI surveillance from four states participating in the Centers for Disease Control and Prevention\u2019s Emerging Infections Program. The analysis included specimens collected in all settings and was limited to case patients who were identified as having fee-for-service Medicare and Part D drug coverage for the year preceding specimen collection. Inpatient hospitalization data was extracted from Medicare Provider Analysis and Review (MEDPAR) files and outpatient AU (prescriptions filled) was determined using Part D drug event files. Timing of hospitalizations and antibiotic prescriptions were described as recent (0\u20133 months prior to specimen collection) or remote (4\u201312 months prior).We studied incident CDI cases filled \u22651 course of outpatient antibiotics in the prior year; 805 (41%) filled an antibiotic both recently and remotely, 497 (25%) only remotely, and 292 (15%) only recently.st generation cephalosporins (10%), and folate pathway inhibitors (10%).Cases with outpatient AU received a median of 23.5 (IQR 12\u201346) total days supplied, and a median of 2 different antibiotic classes (IQR 1 \u2013 3). The most frequent antibiotic classes filled include fluoroquinolones , 1Overall, 1,314 (67%) cases were hospitalized in the prior year; 569 (29%) were hospitalized both recently and remotely, 446 (23%) only recently, and 299 (15%) only remotely. Median length of stay was 13 days (IQR 6\u201328).A total of 142 cases (7%) did not have hospitalization or outpatient AU in the prior year, and 1,097 (56%) had both.Incident CDI cases have substantial exposure to recent and remote hospitalization and outpatient AU. Understanding cumulative effects of multiple risk factors can guide prevention strategies, including antibiotic stewardship efforts.Scott Fridkin, MD, Pfizer: Grant/Research Support Ghinwa Dumyati, MD, Pfizer: Grant/Research Support."} +{"text": "Introduciton: Repetetive transcranial magnetic stimulation (rTMS) is an effective and safety noninvasive technique for treatment of major depression disorder (MDD). There is a body of increasing evidences on the potential molecular mechanisms underlying its effectivity even in case of treatment resistant depression (TRD), however, the exact mechanism is still not clarified. Among multiple biological systems, inflammation can be a target of rTMS in MDD .Here we analysed serum cytokine levels in TRD before and after rTMS interventions.We used bilateral stimulation (15Hz for left DLPC and 1Hz on the right side) in 18 patients with TRD for 2x5 days. Blood samples were collected before the first (V1) and after the last inetrevntion (V2). Phenotypic changes were measured by Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI), Snaith\u2013Hamilton Pleasure Scale (SHAPS), Insomnia Severity Index (ISI) and Stroop Color-Word Test (SCWT) modified by Golden. Inflammatory cytokines were assessed by ELISA assays.Change of BDI and BAI scores between V1 and V2 is associated with difference of TNFalpha levels . Decrease on SHAPS score has been depended on IL-6 level (p=0.027) and the interaction of TNFalpha and IL-10 . Sleep disturbance and neurocognitive function was not associated with cytokine levels.Our results confirmed the association between depressive, anxious and anhedonia symptom improvement and inflammatory mechansims during rTMS treatment. The study was supported by the OTKA 151513 grant."} +{"text": "High prevalences of HIV and other sexually transmitted infections (STIs) have been reported in the current global monkeypox outbreak, which has affected primarily gay, bisexual, and other men who have sex with men (MSM) had received an HIV diagnosis, 816 (41%) had another reportable STI diagnosed in the preceding year, and 363 (18%) had both; 1,208 (61%) persons had either Table 2Table 2.*Among persons with monkeypox, the weekly percentage with concurrent HIV infection increased over time (31%\u201344% by July). The percentage of persons with monkeypox who had HIV infection was higher in older age groups: among persons aged 18\u201324 years, HIV prevalence was 21%, and among those aged \u226555 years, was 59%. HIV prevalence among persons with monkeypox also varied by race and ethnicity, ranging from a high of 63% among non-Hispanic Black or African American (Black) persons, to 41% among Hispanic or Latino (Hispanic) persons, 28% among non-Hispanic White persons, and 22% among non-Hispanic Asian persons.\u03bcL. The median interval since HIV diagnosis was 10 years (IQR\u00a0=\u00a06\u201315 years). Data on HIV PrEP use were available for 172 (14%) persons without HIV infection, 115 (67%) of whom reported current PrEP use.Among 755 persons with monkeypox and HIV infection, 713 (94%) received HIV care in the preceding year, 618 (82%) were virally suppressed, and 586 (78%) had CD4 count \u2265350/\u03bcL, those with CD4 counts <350/\u03bcL more commonly experienced fever (69% versus 59%) and generalized pruritis (53% versus 42%).Compared with persons with monkeypox who did not have HIV infection, those with HIV infection were more likely to report rectal pain (34% versus 26%), tenesmus (20% versus 12%), rectal bleeding (19% versus 12%), purulent or bloody stools (15% versus 8%), and proctitis (13% versus 7%), but were less likely to report lymphadenopathy (48% versus 53%) . The pre\u03bcL, nine (15%) were hospitalized.Among 1,308 (66%) persons with information on hospitalization, the proportion of persons hospitalized with monkeypox was lower among those without HIV infection than among those with HIV infection . Among 45 persons with monkeypox and HIV infection who were not virally suppressed, 12 (27%) were hospitalized, and among 61 with a CD4 count <350 cells/Monkeypox virus and HIV coinfection among Black persons (63%), Hispanic persons (41%), and persons aged \u226555 years (59%) to overall HIV prevalences among Black MSM (39%), Hispanic MSM (19%), and MSM aged 50\u201360 years (32%), respectively exceeded the overall percentage of persons with diagnosed HIV infection who received care in 2020 (74%) , supporting differences in site of exposure as a likely explanation. In a prospective cohort in Spain, MSM with monkeypox who engaged in receptive anal sex were more likely to report proctitis and systemic signs and symptoms preceding rash are highly prevalent among persons with monkeypox.Among 1,969 persons with monkeypox in eight U.S. jurisdictions, 38% had HIV infection, and 41% had an STI in the preceding year. Among persons with monkeypox, hospitalization was more common among persons with HIV infection than persons without HIV infection.It is important to leverage systems for delivering HIV and STI care and prevention and prioritize persons with HIV infection and STIs for vaccination. Screening for HIV and other STIs and other preventive care should be considered for persons evaluated for monkeypox, with HIV care and HIV preexposure prophylaxis offered to eligible persons."} +{"text": "Long-acting injectable (LAI) antipsychotics are related to proven compliance to treatment and more constant medication levels (hence the apparent lower side-effect burden).To highlight the experience with LAI antipsychotic treatment during pregnancy.Literature review.Seven cases are reported. A 35year old with schizophrenia received zuclopenthixole LAI (mostly 200mg/monthly) during both her pregnancies . A 35year old with schizophrenia was under risperidone LAI (25mg/2 weeks) and gave birth to a healthy girl at week 37. Another 35year old (probably with schizophrenia) was on olanzapine LAI (300mg/month during the last quarter of her pregnancy) that led to the birth of a healthy girl at week 40. A 37year old with schizophrenia received paliperidone LAI and gave birth to a healthy boy at week 39. Paliperidone LAI (50mg/monthly) was the treatment of another 34year old with schizoaffective disorder that gave birth to a healthy boy at week 40, as well as of a 26year old (263mg/3-monthly), mother of a healthy boy as well (born at an unspecified week of pregnancy). Finally, a 43year old with bipolar disorder was on aripiprazole LAI (300mg/monthly) during her pregnancy that led to the birth of a healthy girl at week 40.All pregnant women on LAI antipsychotic treatment gave birth to (apparently) healthy babies. LAI doses were mostly low. Long-term follow-up could clarify eventual delayed aftereffects. Based on the literature, LAI antipsychotic treatment could be considered as an option for selected pregnant patients.No significant relationships."} +{"text": "The toxic effects of salinity on agricultural productivity necessitate development of salt stress tolerance in food crops in order to meet the escalating demands. Plants use sophisticated epigenetic systems to fine-tune their responses to environmental cues. Epigenetics is the study of heritable, covalent modifications of DNA and histone proteins that regulate gene expression without altering the underlying nucleotide sequence and consequently modify the phenotype. Epigenetic processes such as covalent changes in DNA, histone modification, histone variants, and certain non-coding RNAs (ncRNA) influence chromatin architecture to regulate its accessibility to the transcriptional machinery. Under salt stress conditions, there is a high frequency of hypermethylation at promoter located CpG sites. Salt stress results in the accumulation of active histones marks like H3K9K14Ac and H3K4me3 and the downfall of repressive histone marks such as H3K9me2 and H3K27me3 on salt-tolerance genes. Similarly, the H2A.Z variant of H2A histone is reported to be down regulated under salt stress conditions. A thorough understanding of the plasticity provided by epigenetic regulation enables a modern approach to genetic modification of salt-resistant cultivars. In this review, we summarize recent developments in understanding the epigenetic mechanisms, particularly those that may play a governing role in the designing of climate smart crops in response to salt stress. Unpredictable climatic conditions render plants suffer from an array of abiotic stress factors. Soil salinity is a key stressor impeding crop productivity and affects an area of more than one billion hectares all over the world and these numbers are constantly growing .At molecular level, plants respond to an environmental stress by implementing dynamic changes in gene expression and reprogramming the plant physiology . In the Methylation of DNA is the most extensively investigated epigenetic modification and includes the insertion of a methyl group at 5\u2032 position on cytosine bases or 6\u2032 position of the adenine bases . DNA metIn order to counter unfavorable environmental conditions, histone protein sustain some modifications at their N\u2032 termini to modulate the gene expression for better survival. It has now been documented that histone acetylation and methylation are vital epigenetic marks in fine tuning gene expression under unfavorable conditions . H3K4me3Plant DNA methylation is referred to as N6-methyladenine (6\u00a0mA) or 5-methylcytosine (5\u00a0mC) . HoweverArabidopsis thaliana. Salt stress has been shown to affect methylation in different ways in different plant species and modify gene expression and thus endowing salt tolerance .OsBZ8 gene expression due to significant differences in chromatin modification between Oryza sativa varieties IR64 and Nanabokra under salt stress. It has been demonstrated that tempering histone proteins provide an epigenetic molecular apparatus for priming plants to salt stress via the modulation of crucial salt responsive genes perpetuated throughout vegetative growth mediated H3 acetylation of polyploidy wheat genes involved in ROS production has been reported to up-regulate transcriptional changes of these genes in response to salt stress gene families encoded by 12 HAT genes (ZmEXPANSIN B2 and ZmXYLOGLUCAN endotransglucosylase/hydrolase1) are up-regulated due to increased H3K9 acetylation at both the promoter and coding regions of genes. The increased acetylation of these genes is attributed to increased mRNA expression of two HAT genes (ZmHATB and ZmGCN5) under salt stress protein involved in cell wall biosynthesis and salt tolerance to play decisive role in strengthening the salt tolerance in bread wheat. Further understanding the defined mechanisms by which HATs activities are modulated will offer new insight into the complex network regulating plant adaptation and tolerance to stress.Although some HAT gene expression levels are shown to increase H4K5 acetylation during salt stress conditions, certain Histone deacetylases respond negatively to the salt stress resistance. Similarly in rice . Zheng eA. thaliana genome are encoded by 18 genes. Studies documented that upon exposure to abiotic stress, histone deacetylase genes display diversified responses and play a crucial role in how plants behave under such conditions Reduced potassium dependency 3 (RPD3)-like, ii) Silent Information Regulator 2 (SIRT) and iii) HD-tuins. The three Histone deacetylase families in the nditions .OsHDA1 was reported to negatively affect the transcriptional activation of OsSOS1 in rice and late embryogenesis abundant protein1 (LEA1) genes, which are essential for salt tolerance in rice, by decreasing H3 acetylation in the promoter regions of LEA1 and SOS1 genes (Arabidopsis class I (HDA19) family histone deacetylases are implicated in positive salinity responses and class II (HDA5/14/15/18) reduced potassium dependency3 (RPD3) histone deacetylases are involved in negative salinity responses and decrease histone H3 lysine 9 dimethylation and/or decreases histone H3 lysine 9 dimethylation (H3K9me2) associated with salt responsive genes and active marks and IR64 (JMJ15 gene (coding for H3K4 demethylase) over expression in A. thaliana under salt stress radically improved salt tolerance . These oolerance . The effolerance . AlthougThe histone methylation and acetylation have been extensively investigated in different plant species under salt stress conditions. Investigations deciphering other histone modifications may enrich our knowledge about other important epigenetic marks and their exploitation for breeding climate smart crops.Oryza sativa and A. thaliana have distinct variants. H2A is the most widely investigated histone and consists of H2A, H2A.Bbd, H2A.X, and H2A.Z variants . Similarr stress . H2A.Z hr stress during sve genes . Accumulde novo DNA methylation and gene silencing using 24- nucleotide small-interfering RNAs and long non-coding RNAs in the RNA-directed DNA methylation process transcription factors under salt stress is transcriptionally regulated mainly by RdDM pathway under salt stress in A. thaliana. 24-nt siRNAs target a region approximately 500bp upstream of the transcription start site of MYB74, which is heavily methylated. Levels of DNA methylation in this region were significantly diminished in wild type plants under salt stress, whereas no changes were observed in RdDM mutants. These observations suggest that changes in the levels of the five 24-nt siRNAs regulate the MYB74 transcription factor via RdDM under salt stress conditions (A. thaliana (H. vulgare (Spirodela polirhiza (nditions . The salnditions . The invthaliana , H. vulg vulgare , cotton vulgare , Spirodeolirhiza and sorgolirhiza .Z. mays displayed down-regulation of miR-250, miR-205, miR-330 and miR-17 in leaves and roots (casein kinase II, GPX, P5CS, IF-1 and some other genes essential for better survival of the plant under saline conditions. This is how miRNAs regulate gene expression under stress conditions and help plants in their survival under harsh environmental conditions. Apart from 24\u00a0nt long miRNAs, the long non-coding RNAs abbreviated as lncRNAs have also been defined as riboregulators longer than 200 bp (Glycine max roots under salt stress conditions. For example, the long non-coding RNA NPC60 expression was escalated 100 times under salt stress condition. Similarly salt treatments enhanced levels of long non-coding RNA973 in cotton (Under salt stress conditions nd roots . Down-ren 200 bp . They aln cotton . The oveMany findings have emphasized epigenetic regulations as powerful mechanisms for regulating the implications of salt stress on plants and provide an excellent foundation for development of salt-tolerant crop plants. In plants susceptible to salt stress, epigenetic controls are associated with the stringent control of gene expression. Epigenetic marks on stress-induced genes dynamically affect the accessibility of chromatin and the expression of those genes. The different regulatory mechanisms for abiotic stress responses might involve epigenetic alterations such as methylation, histone changes, chromatin remodelling, histone variants and lncRnAs.The critical role of epigenetic modifications in regulating gene expression and their ability to transfer to the next generation makes them a unique adaptation tool for plants. The phenotypic plasticity caused by epigenetic variation, which in turn, is through changes in gene expression, will affect fitness and eventually natural selection in plants. Unlike classic DNA sequence mutations, epimutations can happen at much shorter times, and even though they are stable, they are primarily reversible, making them a perfect tool for a quick emergency response to unpredictable environmental stresses. It must also be highlighted that epigenetic changes are typically dependent on the underlying genetic variation, and these two factors must be addressed concurrently. Future study is required to better understand the epigenetic mechanisms behind chromatin changes and the resulting transcriptional regulation that impacts plant responses to environmental stresses. More study on the mechanism of hereditary stress memory is also required."} +{"text": "The genomic integrity of our cells is critical to their normal function, and is protected though the activity of many diverse and essential signalling pathways, with dysregulation of these pathways leading to increasing levels of genomic instability . GenomicTumor Hypoxia Drives Genomic Instability by Tang et al. explores the mechanisms associated altered and dysregulated in the hypoxic tumour environment. The paper focuses on how tumour hypoxia induces genome instability, though activation and modulation of DNA damage responses . The paper also explores the effects of hypoxia on therapeutic treatment responses, and thier manipulation to maximize therapeutic effects of current and future treatments.The review The Last Chance Saloon by Hong et al. reviews the consequences of chromatin bridges on chromosome segregation, cellular replication and genomic instability. They detail the processes leading to chromatin bridge formation, the cellular responses when chromatin bridges are detected (abscission checkpoint activation), and the how chromatin bridges are processed (by the TREX1 exonuclease and LEM-3/ANKLE1 endonucleases). They highlight the role and elements of the NoCut checkpoint involved in protecting genome stability though the management of chromatin bridges, and how father investigation of these nucleases may be relevant of many solid tumour types.Chromatin bridges can form from alterations in DNA metabolism (including chromosome mis-segregation) are resolved before cells division. Chen et al.Recent Advances in the Role of Discoidin Domain Receptor Tyrosine Kinase 1 and Discoidin Domain Receptor Tyrosine Kinase 2 in Breast and Ovarian Cancer concentrates on discussing the role of the transmembrane Discoidin domain receptor tyrosine kinases (DDRs). The review discusses the activation of the kinase activity DDR1 and DDR2 to regulate MAPK signaling, Notch signaling pathways and alter the tumour microenvironment influencing cell invasion and metastasis. The authors highlight the role of DDR1 and DDR2 in breast and ovarian tumour development and progression, and how their dysregulation can alter treatment responses.The review by RAD51AP1 and RAD54L can underpin two distinct RAD51-dependent routes of DNA damage repair via homologous recombination by Selemenakis et al. identifies and explores differential roles for RAD51AP1 and RAD54L in homologues recombination, though RAD51-dependent signalling mechanisms. They reveal the existence of the RAD51AP1- and RAD54L-dependent HR sub-pathways, and show that RAD51AP1 can compensate for RAD54L loss. Importantly, the demonstrate that cell deficient in RAD51AP1 and RAD54L are sensitized to the PARP inhibitor Olaparib.Homologues recombination (HR) is a high-fidelity mechanism for protecting genome integrity from double strand breaks . HR is fLikhatcheva et al. used a combination of a Tip60-targeted inhibitor (TH 1834) (A Novel Mechanism of Ataxia Telangiectasia Mutated Mediated Regulation of Chromatin Remodeling in Hypoxic Conditions. They found ATM activation (pS 1981) under hypoxic stress does require Tip60 activity, in a H3K9me3 positive heterochromatic state. Under these hypoxic conditions, activated ATM regulated H3K9me3 levels through the downregulation of MDM2, which protects Suv39H1 levels (facilitating Suv39H1-dependent H3K9me3). This work reveals the importance of understanding changes to genomic integrity signaling cascades in a hypoxic environment (which better reflects the intra-tumour environment), which will inform new anti-cancer treatment strategies and options.Ataxia Telangiectasia Mutated (ATM) is a key regulator of the DNA double strand break response (DDR), protecting genome integrity against DNA double strand breaks . ImportaTH 1834) and siRNThe E3 Ubiquitin Ligase NEDD4L Targets OGG1 for Ubiquitylation and Modulates the Cellular DNA Damage Response by Hughes and Parsons investigated the role of OGG1 (8-Oxoguanine DNA glycosylase) in protecting genome stability through the base excision repair (BER) pathway. The BER pathway protects the genome from reactive oxygen is 8-oxoguanine (8-oxoG) induced lesions, which can impair DNA replication and genomic integrity induced oxidative stress, which enriched OGG1 levels, decreasing irradiated cells survival while conversely increasing their DNA repair capacity. This suggests that OGG1 mediates the formation intermediate DNA lesions which are reduce cellular survival. This work reveals how OGG1 protein mediates BER, maintaining genome stability and influencing cell survival.The research article ntegrity . Here thThis collection of research papers and reviews highlight the importance of how understanding the intrinsic cellular environment impacts on genome integrity, by mediating the choice and function of DNA repair pathways. This is illustrated by the research papers exploring the effects of hypoxia on genome integrity signalling. We anticipate this collection will be of interest to both researchers and clinician scientists, and highlights new avenues and targets for therapeutic development as anti-cancer treatments."} +{"text": "Veno-venous extracorporeal membrane oxygenation (ECMO) provides blood oxygenation and carbon dioxide removal in acute respiratory distress syndrome. However, during ECMO support, the native lungs still play an important role in gas exchange, functioning as a second oxygenator in series with ECMO. The hypoxic vasoconstriction mechanism diverts regional blood flow within the lungs away from regions with low oxygen levels, optimizing ventilation/perfusion matching. ECMO support has the potential to reduce this adaptive pulmonary response and worsen the ventilation/perfusion mismatch by raising venous oxygen partial pressure. Thus, the objective of this study was to evaluate the effect of ECMO on regional pulmonary perfusion and pulmonary hemodynamics during unilateral ventilation and posterior lung collapse.Five Agroceres pigs were instrumented, monitored and submitted to ECMO. We used the Electrical Impedance Tomography (EIT) to evaluate lung ventilation and perfusion in all protocol steps. Effects of ECMO support on pulmonary hemodynamics and perfusion involving two different scenarios of ventilation/perfusion mismatch: (1) right-lung selective intubation inducing collapse of the normal left lung and (2) dorsal lung collapse after repeated lung lavage. Data including hemodynamics, respiratory, lung perfusion/ventilation, and laboratory data over time were analyzed with a mixed generalized model using the subjects as a random factor.The initiation of ECMO support provided a significant reduction in Mean Pulmonary Artery Pressure (PAPm) in both situations of ventilation/perfusion mismatch. However, distribution of lung perfusion did not change with the use of ECMO support.We found that the use of ECMO support with consequent increase in venous oxygen pressure induced a significant drop in PAPm with no detectable effect on regional lung perfusion in different scenarios of ventilation/perfusion mismatch.The online version contains supplementary material available at 10.1186/s40635-022-00442-x. The main objective of this therapy is to allow the application of protective mechanical ventilation in patients with severe acute respiratory distress syndrome (ARDS) and, at the same time, to ensure adequate gas exchange . Continuous data of hemodynamic, respiratory, lung ventilation/perfusion, and laboratory data over time were analyzed using a mixed generalized model with the subjects as a random factor. We included five animals in the study and all data regarding lung ventilation and perfusion were available for analysis. We were able to induce lung collapse with selective intubation of the right lung, which led to near zero ventilation to the left lung. Perfusion to the collapsed lung was significantly reduced. Left lung collapse was also associated with a significant increase in mean pulmonary arterial pressure and in the fraction of pulmonary shunt , we noticed a reduction in ventilation to dorsal regions secondary to lung collapse. Although this finding remained consistent throughout the 3 stages, we found an increase in posterior lung ventilation from Stage 8 to 10 due to progressive natural lung recruitment , we showed that left lung perfusion was significantly reduced after complete left lung atelectasis . Deoxygenated blood returning from peripheral compartment pass partially through ECMO system. However, since there is no gas flow in the circuit, the blood still returns to the circulation deoxygenated and gas exchange still occurs exclusively on native lungs. Figure S3. Schematic drawing representing experimental model after ECMO canulation and with gas flow (Gas Flow ON). Deoxygenated blood returning from peripheral compartment pass partially through ECMO system. Gas exchange occurs in the ECMO system and blood returns oxygenated to the right side of the heart. The heart pumps the blood to the native lung in which gas exchange also occurs (acting as two oxygenators in series). The oxygenated blood return to the left side of the heart and is posteriorly pumped to the peripheral compartment. Figure S4. (A) : Lung ventilation before and after selective intubation with left lung atelectasis without ECMO circuit. Left lung ventilation was reduced to near zero values and returned to normal values after returning bilateral ventilation. (B): Left lung perfusion before and after unilateral ventilation with left lung atelectasis without ECMO circuit. Left lung perfusion significantly decreased after selective intubation and returned to previous values after returning to bilateral ventilation. (C): Pulmonary shunt before and after unilateral ventilation with left lung atelectasis without ECMO circuit. Pulmonary shunt significantly increased after selective intubation and returned to previous values after returning to bilateral ventilation. (E): PAPm = Mean Pulmonary Artery Pressure. PAPm before and after unilateral ventilation with left lung atelectasis without ECMO circuit. PAPm significantly increased after selective intubation and returned to previous values after returning to bilateral ventilation. Data was not homogenous to all animals. Table S1. Comparison between stages Baseline, Stage 1 and Stage 2. Before installation of ECMO circuit in two different scenarios: Bilateral ventilation and unilateral ventilation. PAC SvO2 denotes venous oxygen saturation acquired in the Pulmonary Arterial Catheter; PaO2 and PvO2 denotes arterial and venous oxygen partial pressure, respectively; PaCO2 denotes arterial carbon dioxide partial pressure; SaO2 denotes arterial oxygen saturation and BE denotes Base Excess. Table S2. Comparison between stages 3 (S3) and 4 (S4). Variables without (Gas Flow OFF) and with ECMO support (Gas Flow ON). PAC SvO2 denotes venous oxygen saturation acquired in the Pulmonary Arterial Catheter; PaO2 and PvO2 denotes arterial and venous oxygen partial pressure, respectively; PaCO2 denotes arterial carbon dioxide partial pressure; SaO2 denotes arterial oxygen saturation and BE denotes Base Excess. Figure S5. (A):PvO2=Venous Oxygen partial pressure. PvO2 significantly increased after initiation of ECMO support. (B): PAPm=Mean Pulmonary Artery Pressure. PAPm did not change after initiation of ECMO support during bilateral ventilation and no previous hypoxemia. (C): Left/Right Lung perfusion before and after initiation of ECMO support during bilateral ventilation. ECMO support by itself did not promote any variation on lung perfusion. (D): Anterior/Posterior lung perfusion before and after initiation of ECMO support during bilateral ventilation. ECMO support by itself did not promote any variation on lung perfusion. Table S3. Comparison between stages 3 (S3) and 5 (S5). Bilateral ventilation (S3) and Unilateral Ventilation (S5) after blood circulation through ECMO circuit. PAC SvO2 denotes venous oxygen saturation acquired in the Pulmonary Arterial Catheter; PaO2 and PvO2 denotes arterial and venous oxygen partial pressure, respectively; PaCO2 denotes arterial carbon dioxide partial pressure; SaO2 denotes arterial oxygen saturation and BE denotes Base Excess. Figure S6. (A): Left lung ventilation before and after selective intubation and left lung collapse with blood flow through ECMO circuit. There was a significant reduction in left lung ventilation. (B): PvO2=Venous Oxygen partial pressure. PvO2 remained stable after selective lung ventilation and left lung collapse with blood flow through ECMO circuit. (C): Left lung perfusion before and after selective intubation and left lung collapse with blood flow through ECMO circuit. Blood flow through ECMO did not affect the capacity of the EIT to detect the reduction in left lung perfusion."} +{"text": "Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococccus (VRE). Nonclinical and Phase 1 (Ph1) clinical data indicate CZD may cause less myelosuppression, particularly with longer duration therapy, and with reduced risk of monoamine oxidase inhibition compared with linezolid (LZD). In Phase 2 (Ph2) and Phase 3 (Ph3) complicated skin and soft tissue infections (cSSTI) clinical trials, CZD was compared with LZD with dosing for 7-14 days, and noninferiority was demonstrated in the Ph3 study. Overall safety was comparable to LZD, and the most common treatment emergent adverse events (TEAEs) in both the CZD and LZD groups were gastrointestinal; however, hematologic laboratory abnormalities and TEAEs were less common with CZD. In June 2021, CZD was approved in China for cSSTI. Sequential therapy with intravenous contezolid acefosamil followed by CZD PO is being evaluated in global Ph3 diabetic foot infection (DFI) and acute bacterial skin and skin structure infection (ABSSSI) clinical trials. Because DFI requires longer treatment (14-28 days), changes in platelet counts with therapy \u226511 days were evaluated in completed Ph2 and Ph3 CZD cSSTI studies.Contezolid is a novel oral (PO) oxazolidinone with potent activity against Gram-positive pathogens, including methicillin-resistant In Ph2 and Ph3 cSSTI studies, mean percent changes in platelet counts from baseline (BSL) to the end of therapy (EOT) visit were compared for all subjects in the safety analysis sets (SS) who received CZD 800 mg and LZD, and also the subjects who received \u226511 days of therapy.In the Ph2 and Ph3 cSSTI studies, the mean platelet count values for LZD subjects decreased compared with CZD subjects, and the difference was greater in subjects who received \u226511 days of therapy.In Ph2 and Ph3 cSSTI clinical trials with treatment durations of 7-14 days, mean platelet counts did not decrease for CZD subjects while mean values for LZD subjects did decline, consistent with nonclinical and Ph1 clinical data. Differences were more significant in subjects who received \u226511 days of therapy. In the current Ph3 global DFI study which compares 14-28 days of CZA/CZD to LZD, evaluation of hematological safety is an important outcome measure.Edward Fang, MD, MicuRx Pharmaceuticals Inc: Employee Huahui Yang, MS, MicuRx Pharmaceuticals Inc: Employee."} +{"text": "In 1988, the World Health Assembly established the Global Polio Eradication Initiative (GPEI). Since then, wild poliovirus (WPV) cases have decreased approximately 99.99%, and WPV types 2 and 3 have been declared eradicated. Only Afghanistan and Pakistan have never interrupted WPV type 1 (WPV1) transmission. This report describes global progress toward polio eradication during January 1, 2020\u2013April 30, 2022, and updates previous reports to bivalent OPV in all OPV-using countries, bOPV and injectable inactivated poliovirus vaccine (IPV) have been used in routine immunization programs worldwide. cVDPV type 2 (cVDPV2) has been the predominant cause of cVDPV outbreaks since 2006 and informed the rationale for the switch to bOPV. Monovalent OPV Sabin type 2 (mOPV2) is reserved for cVDPV2 outbreak response campaigns , 201 million mOPV2, and 51 million tOPV doses through 145 supplementary immunization activities (SIAs)In November 2020, the World Health Organization (WHO) granted Emergency Use ListingThe primary system for detecting poliovirus is case-based syndromic surveillance for acute flaccid paralysis (AFP), with confirmation by stool specimen testing done at one of 146 WHO-accredited laboratories across 92 countries, comprising the Global Polio Laboratory Network. The two primary indicators used to assess surveillance performance include the nonpolio AFP (NPAFP) rateWhether or not AFP surveillance performance indicator targets are met, gaps often exist in poliovirus detection subnationally. These gaps can be addressed through environmental surveillance (ES), the systematic collection and testing of sewage samples for poliovirus .Countries reporting WPV cases and isolations. In 2021, five WPV1 cases were reported from the two remaining countries with endemic polio: four from Afghanistan and one from Pakistan compared with 2020 (untries) . Thirty-After the last identified indigenous WPV1 case in Nigeria in 2016, the WHO African Region was certified WPV-free in August 2020. In 2021, the region reported its first case of WPV1 in approximately 5 years. In the absence of sustained transmission, this single case does not change the Africa Region\u2019s WPV-free status. Afghanistan and Pakistan continue to have endemic WPV1 circulation; thus, only one WHO region (the Eastern Mediterranean Region) is not certified WPV-free. Although substantial improvements in eradication activities have been made in both countries, insecurity, instability, mass population movements, and vaccine refusal continue to pose challenges. COVID-19 pandemic prevention efforts have affected AFP surveillance sensitivity and the administration of routine childhood immunizations globally transmission remains endemic in Afghanistan and Pakistan. Outbreaks of paralysis due to circulating vaccine-derived polioviruses (cVDPVs) occur in populations with low immunity following prolonged circulation of Sabin strain oral poliovirus vaccine.In 2021, Afghanistan and Pakistan reported a sharp decline in WPV1 cases from previous years. A WPV1 case genetically linked to these countries occurred in Malawi in November 2021.Current progress toward polio eradication must be sustained in countries experiencing endemic transmission and outbreaks. Intensified programmatic actions leading to more effective outbreak responses and enhanced efforts to immunize all children are essential. Until WPV1 is eradicated and cVDPV transmission is interrupted, the risk for children being paralyzed by polio remains."} +{"text": "Hemiptera exhibited the highest species diversity with 20 species. Gyeonggi-do was where the highest number of invasive alien insect species were identified (45 species). Species richness analysis revealed that Jeju-do showed the highest Dominance Index (0.8), whereas Gyeongsangnam-do had the highest Diversity Index (2.8). Corythuchamarmorata (Hemiptera: Tingidae), Lycormadelicatula (Hemiptera: Fulgoridae), Ophraellacommuna (Coleoptera: Chrysomeridae), Metcalfapruinosa (Say) (Hemiptera: Flatidae) and Pochaziashantungensis (Hemiptera: Ricaniidae) were distributed in more than 300 locations of the country. Invasive alien insect species inhabited the roadsides (31.3%), farmlands (18.3%) and parks (16.6%). In this study, we list the invasive alien insect species in Korean ecosystems and provide a basis for selecting primary management target species.We investigated the identity and distribution of the invasive alien insect species inhabiting Korean ecosystems, targeting 3,249 locations in nine regions between 2015 and 2018. In natural ecosystems, we identified 63 species in 43 families and nine orders of invasive alien insect species, respectively. We observed that the order Amongst these, the 9th goal states that the introduction and settlement of invasive alien species should be prevented by identifying their introduction routes and destroying them.Increasing cross-border exchanges and international trade are contributing to the rapid introduction of invasive alien species (IAS) globally , which, An invaded ecosystem may not have predators or competitors that would limit the growth of the invasive alien insect species . AlternaThe unintentional introduction of IAS can occur via ships or on freights or even Lycormadelicatula (Hemiptera: Fulgoridae), Metcalfapruinosa (Hemiptera: Flatidae) and Pochaziashantungensis (Hemiptera: Ricaniidae) are widespread in Korea, where they cause severe ecological damage to crops and trees by piercing-sucking (Vespavelutinanigrithorax (Hymenoptera: Vespidae), which is also widespread in Korea, preys on honeybees, decreasing the earnings of beekeepers and it also harms humans are listed amongst the International Union for Conservation of Nature\u2019s (IUCN) 100 most common invasive species is a large insect species that competes with any native Korean insect; as it presents diverse feeding habits, it may damage agricultural or forest land , Diversity Index (H'), Richness Index (RI) and Evenness Index (EI) for 3,249 locations of the nine regions.DI was calc\\documentclass[12pt]{standalone}\\usepackage{varwidth}\\usepackage[utf8x]{inputenc}\\usepackage[T1]{fontenc}\\usepackage{lmodern}\\usepackage{amsmath, amssymb, graphics, setspace}\\newcommand{\\mathsym}[1]{{}}\\newcommand{\\unicode}[1]{{}}\\newcounter{mathematicapage}\\begin{document} \\begin{varwidth}{50in} \\begin{equation*} (n1+n2)/N \\end{equation*} \\end{varwidth}\\end{document}DI = where n1 is the number of dominant species, n2 is the number of subdominant species and N is the total number of individuals.H' represents the relationship between species and the number of individuals in a population. It is a measure of species enrichment and uniformity, correlating positively with the species diversity in a population. The following Shannon-Wiener function (H\u2019), developed by \\documentclass[12pt]{standalone}\\usepackage{varwidth}\\usepackage[utf8x]{inputenc}\\usepackage[T1]{fontenc}\\usepackage{lmodern}\\usepackage{amsmath, amssymb, graphics, setspace}\\newcommand{\\mathsym}[1]{{}}\\newcommand{\\unicode}[1]{{}}\\newcounter{mathematicapage}\\begin{document} \\begin{varwidth}{50in} \\begin{equation*} -\u2211(i=1)^s[(ni/n) ln(ni/n)] \\end{equation*} \\end{varwidth}\\end{document}H\u2019 = where ni is the number of species and N is the total number of locations.RI represents the state of a population, based on the number of species and the total number of locations. The higher RI is, the better the environment is in terms of species richness. We calculated RI using \\documentclass[12pt]{standalone}\\usepackage{varwidth}\\usepackage[utf8x]{inputenc}\\usepackage[T1]{fontenc}\\usepackage{lmodern}\\usepackage{amsmath, amssymb, graphics, setspace}\\newcommand{\\mathsym}[1]{{}}\\newcommand{\\unicode}[1]{{}}\\newcounter{mathematicapage}\\begin{document} \\begin{varwidth}{50in} \\begin{equation*} (S-1)/(ln (N)) \\end{equation*} \\end{varwidth}\\end{document}RI = where S is the number of species and N is the total number of locations.EI represents the species uniformity in a population and is the ratio of the actual index number over the maximum number of the corresponding index. If all the species\u2019 populations are found in the same number of locations, the maximum EI equals one. We calculated EI using the \\documentclass[12pt]{standalone}\\usepackage{varwidth}\\usepackage[utf8x]{inputenc}\\usepackage[T1]{fontenc}\\usepackage{lmodern}\\usepackage{amsmath, amssymb, graphics, setspace}\\newcommand{\\mathsym}[1]{{}}\\newcommand{\\unicode}[1]{{}}\\newcounter{mathematicapage}\\begin{document} \\begin{varwidth}{50in} \\begin{equation*} H'/lnS \\end{equation*} \\end{varwidth}\\end{document}EI = where H\u2019 is the DI and S is the total number of species.For the 17 species that inhabit the most in Korea, the distribution point is indicated on the map. Using Arc GIS (ver. 10.5), the distribution of 17 species distributed in all nine regions and appear at more than 20 locations is shown on the shapefile map of the Korean Peninsula.Hemiptera exhibited the highest species diversity, with 20 species in 1,972 locations, followed by 12 species of Coleoptera in 676 locations, 12 species of Lepidoptera in 268 locations, five species of Diptera in 86 locations, five species of Blattodae in 97 locations and four species of Hymenoptera in 133 locations . As for the H', the Gyeongsnagnam-do region showed the highest value (2.8). As for the EI, Jeollanam-do, Chungcheongbuk-do and Gyeongsangnam-do showed values higher than (0.9). Finally, regarding RI, Gyeonggi-do showed the highest value (6.5) Table .Corythuchamarmorata appeared in the highest number of locations , followed by farmlands (18.3%), parks 17.5%), forests (16.6%), residential areas (8.0%), orchards (1.7%), watersides (6.3%) and others (0.4%) , forest roads 35.4%), mountainous districts (21.1%) and farm (0.3%). The waterside includes riversides (51.1%), wetlands (35.6%), reservoirs (11.3%) and valleys (2.1%) and the most invasive alien insect species were found on the riverside. Additionally, there are other areas (45.0%), beaches (25.0%), rest areas (20.0%) and camping sites (10.0%) (Fig. .4%, mounFrom 2015 and 2018, we identified 63 species in 43 families and nine orders of invasive alien insect species in the Korean ecosystems. We analysed the distribution and diversity of the invasive alien insect species from nine regions and compared the DI, H\u2019, RI and EI of the species in each region.Depending on the local temperature and geographical impact, invasive alien insect species found between regions may differ. Due to these effects, it seems that the species richness analysis results were different for each region. It seems that the regions with the highest DI, H', RI and EI values appeared differently due to the influence of urbanisation and geographical isolation. The habitats of the invasive alien insect species introduced via various routes can be ranked in descending order as follows: roadsides, farmlands and parks. More than 83% of the invasive alien insect species were found on the roadsides, farmlands and parks. Amongst them, more than 31% of the invasive alien insect species were found on the roadside. This could be explained by the fact that invasive alien insect species are often introduced via transportation of freight and shipping containers , increasSolenopsisinvicta, Anoplolepisgracilipes, Pochaziashantungensis and Vespavelutinanigrithorax are likely to spread across the country (Climate changes are increasing the overall temperature during summer and the lowest temperature during winter in Korea. As temperature directly affects the development, reproduction and survival of insect species , the hig country . TherefoTherefore, the unintentional introduction of invasive alien insect species in Korea must be carefully monitored. Additional investigations are needed to establish standard procedures and to prevent a further spread of the invasive alien insect species in the country. Each region of Korea should also constantly remove and manage the invasive alien insect species that may disturb the ecological equilibrium.List of invasive alien insect species found in the natural ecosystem Table ."} +{"text": "Whilst clinical simulation is established as an effective education tool within the healthcare community, the inability to offer authentic educational learning environments remains problematic. Advances in technology such as immersive virtual reality offer new opportunities to enhance traditional practice to an extent that may transform learning. However, with traditional clinical simulation stress and anxiety can both hinder performance and learning, yet it is unknown what nuances are applicable within a clinical virtual simulation environment. Determining potential benefits, drawbacks (including related stress and anxiety) and affordances of immersive technology clinical simulation designs may help provide an understanding of its usefulness. The aim of this scoping review is to investigate the range and nature of evidence associated with immersive virtual reality clinical simulation and education design. In addition, the review will describe authentic immersive technology clinical simulation use and reported stress response measurements. A search of seven electronic database and grey literature was performed in accordance with the Joanna Briggs Institute methodology. A key term search strategy was employed with five themes identified and investigated: (1) Healthcare professionals, (2) Clinical simulation, (3) Immersive virtual reality, (4) Stress/anxiety and (5) Authentic learning design. Application of the search strategy resulted in a hit total of 212 articles. Twelve articles met inclusion criteria. With most literature focusing on procedural performance and non-transferable education needs, there was a paucity of research that specifically investigated immersive virtual reality clinical simulation education and related stress. Therefore, this scoping review contributes new understandings by providing valuable insight and potential research gaps into current immersive virtual reality clinical simulation, its relationship to stress and the education design models currently being utilised to develop these concepts. Whilst each technology differs slightly, the overarching attributes include layers of sensory information that offer the user a simulated experience of sound, sight, smell and touch similar to the physical world. Users are typically immersed within environments that offer real-world situations and/or representations of virtual humans. This now offers a potential to interact and explore authentic environments in a 360-degree visual format and transcend the boundaries of the physical world All Healthcare professionals, (2) Clinical simulation, (3) IVR, (4) Stress and (5) Authentic learning design. A scoping review of how immersive technology relates to traditional clinical simulation within health care practice will help to define key concepts, map existing research and importantly identify future research gaps within this emerging area of research.The relationship between immersive technology and what impact healthcare clinical simulation has on stress and education is uncertain. To help identify existing gaps in knowledge, the aim of this scoping review is to systematically investigate the range and nature of evidence associated with immersive virtual reality clinical simulation and education design. In addition, the review will describe authentic immersive technology clinical simulation education design and reported stress measurements.https://doi.org/10.17605/OSF.IO/ZP7EC) methodology for scoping review Aiello .Inclusion criteria Table and equiDue to the large number of irrelevant studies from veterinary science and paediatric healthcare, a \u2018human/humans\u2019 and \u2018adult\u2019 limiter was applied. In addition, to ensure a quality approach non-primary research such as guides, product reviews, reports and opinion papers were excluded. Databases were searched from the earliest available date to the 1st November 2021 and included for pragmatic reasons only those of English language with full text. Finally, there are no cultural, geographical or gender-based exclusion interests in this setting.Question: What is the association between immersive virtual reality technology (Concept), authentic traditional clinical simulation (Context) and healthcare training (Population)?Sub-Question 1: How is immersive virtual reality clinical simulation participant stress response measured?Sub-Question 2: Is authentic clinical simulation healthcare education supported by learning design?All supporting evidence that meets a Population, Concept, Context (PCC) structure was considered for inclusion. The following research questions were formulated within the PCC framework.Participants include qualified clinical personnel or students studying towards a clinical qualification. Those who did not include immersive technology and clinical simulation were excluded. In line with the broad inclusion criteria for a scoping review, alternate technology-based simulation concepts illustrating an authentic clinical learning environment were investigated. This further included psychometric and physiological stress measurement within immersive technology but excluded studies that used technology as a direct substitute without functional change.A preliminary search of MEDLINE, the Cochrane Database of Systematic Reviews and\u00a0JBI Evidence Synthesis\u00a0was conducted and no current or underway systematic reviews or scoping reviews on the topic were identified.This review considered both quasi-experimental and experimental study designs including non-randomized controlled trials, randomized controlled trials, before and after studies and interrupted time-series studies. The types of publications included full-text journal articles and Grey literature: Google scholar, books, theses, conference and symposium presentations. All publications of qualitative, quantitative and mixed-method primary research study types were included.To identify appropriate index terms and keywords, the search strategy was tested in two electronic databases MEDLINE (PubMed) and CINAHL (EBSCO). The author consulted with an experienced librarian and performed a comprehensive search using the title and abstract to identify the full strategy keywords and index terms. The final search strategy for all identified keywords and index terms was adapted for each included database and/or information source. Seven databases were searched on 1st November 2021 and included: ERIC, AMED, PsychINFO (OVID), CHINAHL, MEDLINE (EBSCO), SCOPUS and Web of Science (Appendix 2). Grey literature was searched in Google Scholar, and a review of included source reference lists for similar topics were investigated to identify relevant studies.All relevant citation data were exported from databases to Endnote\u2122 X9 before being forwarded to Covidence\u2122 for peer review. The software enabled the identification of duplicate study results, screening and data extraction. This process follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA-ScR) guidelines Prior pilot testing of the source selection was performed to help refine eligibility criteria and discuss discrepancies. (2) Two independent reviewers (SA and TC) screened titles and abstracts against the inclusion and exclusion criteria followed by (3) a full-text screen. Disagreement between SA and TC was resolved by discussion and consensus or by a third reviewer (CS). The two reviewers (SA and TC) critically appraised the included studies within the Covidence\u2122 customised fields. The Inter-rater reliability was 90% with a Cohen\u2019s Kappa of 0.61.Study characteristicsParticipant characteristicsPsychometric and physiological (Stress) measurementImmersive technologiesClinical simulation characteristicsEducation/learning designOutcome characteristicsKey findingsA standardised data extraction tool was developed a priori in Microsoft Excel 2019 . Two independent reviewers (SA and TC) tested and refined the tool and then extracted the final eligible study data from Covidence\u2122 into the predetermined fields. The data included specific details about the participants, concept, context, study methods and included:Charting of the data was recorded independently by SA and TC with answers and detail later verified for consistency. Any disagreement between SA and TC was resolved through (1) discussion, (2) an additional review or (3) a third reviewer (CS). No data within the eligible studies required further clarification from the author. When we encountered duplicate reporting of the same research study, we selected one report (the most detailed) for full review.The findings of the final review are presented below in a tabular form with narrative synthesis. The narrative will explore the patterns, similarities and relationships to stress and authentic immersive simulation learning within healthcare education. The mapping of the results is reported by providing a visual representation of the data to support breadth, extent and range of activity that aligns to the scoping review objective. Synthesis of data was undertaken by the principal investigator (SA) with validation of findings discussed amongst the review team (TC and CS).The search strategy identified 212 articles (107 database and 105 grey literature). Of these, 104 were excluded by title and/or abstract, 67 did not relate to IVR, 18 did not report healthcare clinical simulation, there were 10 duplicates and 1 study was excluded because we were unable to retrieve it. The remaining 12 articles were deemed eligible for inclusion and subject to this review , and one each in \u201cAustralian Journal of Education Technology\u201d, \u201cMultidisciplinary Digital Publishing Institute (MDPI); Information\u201d, \u201cInstitute of Electrical and Electronic Engineers (IEEE)\u201d, \u201cNurse Education Today\u201d, \u201cComputers, Informatics, Nursing (CIN)\u201d and \u201cClinical Simulation in Nursing\u201d.The research was largely recent, with publications spanning between the 2005 and 2020 period Fig.\u00a0, with boThere was a diverse study design with mixed methods being most common : The majority of articles sampled Paramedic : Virtual clinical simulation techniques included avatars : The technology related to alternate virtual electronic environments utilised multiple or single applications and included augmented virtual reality : The assessment methods to investigate anxiety/stress were heterogenous in nature and included a range of quantitative, qualitative and biometric instruments. The range and diversity of questionnaire methods included the NASA Task load index : Whilst it is acknowledged that education and learning is implied within many of the included articles, only four were explicit in describing an educational design approach. Of note, all four articles described a Design Based Research methodology when describing their work , authentic traditional clinical simulation (Context) and healthcare training (Population). In addition, stress measurement tools and authentic learning designs themes are reported (sub-questions 1 and 2). This scoping review identified 12 studies that addressed these topics: Appendix 4 summarises the article themes and highlights the PCC question, sub-questions, context and outcome/findings. To our knowledge, this is the first review of the literature that explores the five related themes: (1) All Healthcare professionals, (2) Clinical simulation, (3) IVR, (4) Stress and (5) Authentic learning design is being utilised by a range of healthcare disciplines. It is clear that the integration of technology not only relates to the assessment of competency but also to support distance learning, reduce costs and promote effective learning those that performed traditional clinical simulation within an immersive (screen projected) CAVE environment and without being tethered to a computer those that did not measure stress or anxiety relates to those subjected to situational contexts that induce anxiety. The required outcome of VRET is to develop stress inoculation to a context or event without negatively impacting critical thinking or performance Virtual reality {including related terms}/ (1986 hits)Clinical simulation {including related terms}/ (12237 hits)Stress {including related terms}/ (13391 hits)Education {including related terms}/ (4472 hits)1 and 2 /(11 hits)3 and 4 /(84 hits)1 and 5 / (599 hits)3 and 5 / (31 hits)1 and 2 and 3 and 4/ (5 hits)1 and 2 and 3 and 4 and 5/ (0 hits)Ovid ERIC (Abstract: full text/English/human)Health care professional {including related terms}/ (7036 hits)Virtual reality {including related terms}/ (155 hits)Clinical simulation {including related terms}/ (101 hits)Stress {including related terms}/ (3397 hits)Education {including related terms}/ (191 hits)1 and 2 /(42 hits)3 and 4 /(14 hits)1 and 5 / (178 hits)3 and 5 / (5 hits)1 and 2 and 3 and 4/ (0 hits)1 and 2 and 3 and 4 and 5/ (0 hits)Ovid AMED (Abstract: full text/English/human)Health care professional{including related terms}/ (99757 hits)Virtual reality {including related terms}/ (2364 hits)Clinical simulation {including related terms}/ (2764 hits)Stress {including related terms}/ (91702 hits)Education {including related terms}/ (1984 hits)1 and 2 /(500 hits)3 and 4 /(387 hits)1 and 5 / (1647 hits)3 and 5 / (41 hits)1 and 2 and 3 and 4/ (6 hits)1 and 2 and 3 and 4 and 5/ (2 hits)Ovid PsychINFO (Abstract: full text/English/human)Health care professional{including related terms}/ (98591 hits)Virtual reality {including related terms}/ (183600 hits)Clinical simulation {including related terms}/ (317807 hits)Stress {including related terms}/ (1535686 hits)Education {including related terms}/ (38474 hits)1 and 2 /(1533 hits)3 and 4 /(17554 hits)1 and 5 / (33607 hits)3 and 5 / (2108 hits)1 and 2 and 3 and 4/ (31 hits)1 and 2 and 3 and 4 and 5/ (17 hits)EBSCO CINAHL complete, EBSCO MedLine (Abstract: full text/English/human)Health care professional{including related terms}/ (172989)Virtual reality {including related terms}/ (1936022)Clinical simulation {including related terms}/ (4598825)Stress {including related terms}/ (7018726)Education {including related terms}/ (532591)1 and 2 /(2949)3 and 4 /(98204)1 and 5 / (28793)3 and 5 / (2093)1 and 2 and 3 and 4/ (43)1 and 2 and 3 and 4 and 5/ (17 hits)SCOPUS (Abstract: full text/English/human)Health care professional{including related terms}/ (100025 hits)Virtual reality {including related terms}/ (445991 hits)Clinical simulation {including related terms}/ (1929266 hits)Stress {including related terms}/ (3477910 hits)Education {including related terms}/ (96497 hits)1 and 2 /(3800 hits)3 and 4 /(129380 hits)1 and 5 / (38042 hits)3 and 5 / (4178 hits)1 and 2 and 3 and 4/ (90 hits)1 and 2 and 3 and 4 and 5/ (71 hits)Web of Science (Abstract: full text/English/human)"} +{"text": "Attitudes towards dementia and caregiving differ by family caregivers\u2019 racial/ethnic backgrounds. However, there is a gap in the literature on midlife women family caregivers\u2019 attitudes toward Alzheimer\u2019s Disease (AD) and family caregiving. The study purposes were to (1) explore racial/ethnic variations in midlife women family caregivers\u2019 attitudes toward AD and family caregiving and (2) examine the relationships among their attitudes towards dementia and caregiving, quality of life, and physical and psychological symptoms. This cross-sectional study was conducted through an online survey among 36 Whites, 41 African Americans, 40 Hispanics, and 55 Asians. The structured measures consisted of two types of attitudes , health-related quality of life (EQ-5D-5L), and multidimensional symptoms (Midlife Women\u2019s Symptom Index). The data were analyzed using one-way ANOVA and multiple linear regression analyses with SPSS 26. Asian caregivers perceived the care recipients\u2019 symptoms as more bothersome than White caregivers (p = .039). Asian caregivers reported lower levels of behavioral skills and shared responsibility compared with other racial/ethnic groups of caregivers (p < .01). African Americans showed more positive attitudes toward family caregiving compared with Hispanics and Asians (p = .001). The regression analyses indicated that more positive attitudes toward family caregiving were significantly related to a better quality of life and fewer symptoms . Culturally tailored interventions that incorporate caregivers\u2019 attitudes are needed to improve midlife women family caregivers\u2019 quality of life and symptoms."} +{"text": "Chronic hepatitis C virus (HCV) infection affects 71 million individuals, mostly residing in low- and middle-income countries (LMICs). Direct-acting antivirals (DAAs) give high rates of sustained virological response (SVR) in high-income countries where a restricted range of HCV genotypes/subtypes circulate.We studied United Kingdom\u2013resident patients born in Africa to examine DAA effectiveness in LMICs where there is far greater breadth of HCV genotypes/subtypes. Viral genome sequences were determined from 233 patients.Full-length viral genomic sequences for 26 known subtypes and 5 previously unidentified isolates covering 5 HCV genotypes were determined. From 149 patients who received DAA treatment/retreatment, the overall SVR was 93%. Treatment failure was associated primarily with 2 subtypes, gt1l and gt4r, using sofosbuvir/ledipasvir. These subtypes contain natural resistance-associated variants that likely contribute to poor efficacy with this drug combination. Treatment failure was also significantly associated with hepatocellular carcinoma.DAA combinations give high SVR rates despite the high HCV diversity across the African continent except for subtypes gt1l and gt4r, which respond poorly to sofosbuvir/ledipasvir. These subtypes are widely distributed across Western, Central, and Eastern Africa. Thus, in circumstances where accurate genotyping is absent, ledipasvir and its generic compounds should not be considered as a recommended treatment option. We examined hepatitis C virus (HCV)\u2013infected patients born in Africa but resident in the United Kingdom for treatment outcomes for direct-acting antiviral therapy. The unusual HCV subtypes identified in many patients successfully responded to treatment except for 2 subtypes. Direct-acting antivirals (DAAs) capable of clearing chronic hepatitis C virus (HCV) infection from 90%\u201395% of treated individuals are a cornerstone of the World Health Organization (WHO) strategy to eliminate viral hepatitis as a public health concern by 2030 . ConsequWe recently reported a substantial gap in our knowledge of HCV genomic sequences circulating in LMICs across large geographic areas . In addiIn this study, we address the issue of treatment outcomes in response to DAA therapy in HCV-infected individuals originating from Africa but residing in the United Kingdom (UK). We also aimed to increase the number of available complete HCV genomic sequences derived from African patients, as there is a lack of such genetic data in public databases.HCV-infected patients were enrolled into the HCV Research UK cohort at clinical sites in the UK . All patThe next-generation sequencing (NGS) methods for determining HCV viral sequences by metagenomics and target enrichment have been published previously and are http://www.bioinformatics.babraham.ac.uk/projects/fastqc/), and low-quality bases (Phred score <30 or read length <50 nucleotides) were trimmed using Trim Galore! script . Cleaned reads were submitted for de novo assembly using SPAdes ) and, within this group, decompensated disease was found in 42 individuals (32%). In addition, 25 cases (19%) were diagnosed with hepatocellular carcinoma (HCC) . The perUsing a NGS target enrichment approach , 3 gt2 subtypes (and 1 gt2 unassigned isolate), 2 gt3 subtypes, 13 gt4 subtypes, and 2 gt5a sequences; unassigned isolates differed from classified HCV subtypes by at least 15% at the nucleotide level . CompareThe most highly represented countries in each geographic African region for sequence data were Egypt , Nigeria , Democratic Republic of the Congo (DRC)/Republic of Congo , Somalia , and South Africa and 1B. We compared the genotypes/subtypes identified by NGS analysis with those recorded by the clinical site that used commercial kits (such as the INNO-LiPA test [Innogenetics]). We obtained genotype and subtype data for both commercial assays and NGS for 133 and 60 samples, respectively (genotype and subtype). HCV genotype was identical in 124 samples (93%), but the percentage of concordant HCV subtypes was much lower (n = 41/60 samples [68%]). The majority of mismatches corresponded to subtypes that are not typically identified or differentiated in commercial kits (data not shown).We evaluated treatment outcomes for all DAA drug combinations used to treat the cohort, yielding data on 149 patients. This group included 12 patients who had received prior DAA therapy on 1 (n = 11) or 2 (n = 1) occasions. Hence, there was a total of 162 treatment episodes recorded . Two patP = .006) was significantly associated with DAA failure; a higher failure rate was noted also for those with decompensated liver disease , there were 24 treatment episodes with DAA that led to treatment failure in 19 pa28R30M31, that could give resistance to NS5A inhibitors and thereby reduce effectiveness of DAA therapy ). SOF was occasionally used as mono-DAA therapy with ribavirin (RBV) but was mostly prescribed in dual combination with NS5A inhibitors with or without RBV or triple combination (VEL/VOX) . The oveAside from gt1l and gt4r, the SVR for other gt1 and gt4 subtypes was 97%. This includes gt4d, another unusual subtype associated with treatment failure with ombitasvir (OBV)/paritaprevir (PTV)/ritonavir (RTV) in a clinical trial . There wThirteen patients in our cohort who were not successfully cured by initial DAA therapy received retreatment, with 9 achieving SVR. Aside from 1 individual who relapsed on retreatment, the more potent DAA combinations containing GLE/PIB and SOF/VEL/VOX achieved cure in 6 patients. SOF/VEL/VOX retreatment was unsuccessful in 1 individual with gt4r who had In conclusion, our study describes HCV subtypes and viral sequences circulating in countries for which there are very limited data, and complements our recent report describing gt4 subtypes in Uganda and DRC as well as novel gt7 strains . CrucialThe Journal of Infectious Diseases online. Consisting of data provided by the authors to benefit the reader, the posted materials are not copyedited and are the sole responsibility of the authors, so questions or comments should be addressed to the corresponding author.Supplementary materials are available at jiab110_suppl_Supplementary_Table_1Click here for additional data file.jiab110_suppl_Supplementary_Table_2Click here for additional data file.jiab110_suppl_Supplementary_Table_3Click here for additional data file.jiab110_suppl_Supplementary_Table_4Click here for additional data file.jiab110_suppl_Supplementary_Table_5Click here for additional data file.jiab110_suppl_Supplementary_Table_6Click here for additional data file.jiab110_suppl_Supplementary_MethodsClick here for additional data file."} +{"text": "Alzheimer's Disease and Related Dementias (ADRD) management and prevention is a priority for providers and community members. Aligning perceptions regarding resources and screening supports person-centered care while addressing the increased dementia burden attributed to systemic inequity and health disparities. This project aimed to identify needs in lower-income diverse populations.Research Question: What are the differences in perceived service gaps and screening preferences for brain health and ADRD?A convenience sample of 15 providers and 20 community stakeholders (55+) completed a 2021 survey about Richmond's service gaps and screening needs for brain health and ADRD.40% of providers were ADRD focused, 65% of community members reported a memory concern. Overall, providers reported fewer service gaps: Administration on Aging Service Gaps 46.7%/75%, Caregiving gaps 80%/85%, Specialty Health gaps: 53.3%/80%. Clinical ADRD gaps 73.3%/80%, Memory Case Management gaps 86.7%/90%, Memory Social Services gaps 33.3%/90%, Memory Screening gaps 53.3%/85%. Provider and Community stakeholders were more aligned regarding screenings that should be included in a brain health/ADRD management program: Advance Care Planning screening: 80%/80%, Caregiving Status screening: 93.3%/90%, Memory Loss screening: Lifestyle Risk screening: 100%/95%, Cognitive-Comorbidity Risks screening: 100%/75%, Psychosocial Risks screening:100%/100%, Depression screening 100%/100%, Clinical Health screening 73.3%/90%, Substance Use Disorder screening 86%/75%, Sleep Problem screening 93%/75%Although there were areas in which both providers and community members aligned, results show that providers underestimate brain health/ADRD service gaps experienced by the community. Both groups are amenable to comprehensive screening for services. However, providers are interested in more expansive ADRD screeners."} +{"text": "Compared with peers, young sexual and gender minorities (SGM) are at a four to seven-fold increased risk of attempting suicide. Prior epidemiological studies, mainly focusing on monomorbid inequalities and without conducting diagnostic clinical interviews, have been unable to report robust data on psychiatric comorbidities among those who attempted suicide. The purpose of this presentation is to describe the presence of current psychiatric comorbidities among SGM youth and young adults at elevated risk for repeat suicide attempts.A diverse convenience sample of SGM youth and young adults with a lifetime history of suicide attempts and current suicidal ideation was recruited for an open-phase suicide prevention trial in San Diego, CA. At baseline, participants underwent a 15-module DIAMOND interview for adults or computerized K-SADS for minors, and a battery of self-report questionnaires.Among the 31 participants , 27 (87%) participants met criteria for any mood disorder, 24 (77%) for any anxiety disorder, 16 (52%) for any trauma or stress disorder, and 2 (6%) for any psychotic disorder. One (3%) participant did not meet criteria for any psychiatric diagnoses, while five (16%) met criteria for a single and 25 (81%) for multiple diagnoses. The average number of diagnoses was 3.2 . Additionally, 20 (65%) participants met the cut-off for likely ADHD, 20 (65%) for possible borderline personality disorder, and 21 (68%) for likely body dysmorphic disorder, with 11 (35%) within the 90th percentile for reference eating disorder severity.The degree of psychiatric comorbidities in the sample of SGM youth and young adults at elevated risk for suicide was high. Beside direct suicide risk mitigation efforts, suicide prevention programs that target young SGM with a history of attempts should screen for untreated psychiatric disorders.\u2022\u2002LGBTQ+ youth and young adults at elevated risk for repeat suicide attempts experience a high degree of psychiatric comorbity.\u2022\u2002Beyond suicide risk mitigation, LGBTQ+ youth suicide prevention programs should focus on untreated psychiatric comorbidities."} +{"text": "In this efficient and mild pathway, the reaction produces sulfonamide compounds under redox-neutral condition, which is mechanistically different from the nitrogen nucleophilic substitution reactions. Significantly, this transformation intends to utilize the property of visible light-induced azides to generate triplet nitrene and followed coupling with sulfonyl radicals in situ to achieve structurally diverse benzenesulfinamides in good yields.We here have developed an S(O) I has induced abnormal heart development in zebrafish embryos [II could have therapeutic potential for the treatment of metabolic disorders [III has potent in vitro antiproliferative activity against several human cancer cell lines including drug-resistant tumor cells [IV has demonstrated remarkably high anti-bacterial activity against ATCC35218 (Escherichia coli) and ATCC6538 (Staphylococcus aureus) 4]. Alo. AloI ha2\u2013N bonds could heavily rely on the classical nitrogen nucleophilic substitution reactions. The traditional method is the amination of arylsulfonyl chlorides 3+ to produce [Ir(ppy)3]3+* *) through energy transfer which participates in two catalytic cycles: a single electron transfer (SET) process with 2 to obtain [Ir(ppy)3]2+ and a sulfonyl radical A accompanied by proton dissociation, and an energy transfer (EnT) process involving 1, resulting in the loss of N2 and the formation of the triplet nitrene B. The intermediate B was captured by Cu(I) to generate a Cu(III) nitrene intermediate C. After underwent a SET process and protonation, C was converted to a Cu (II) complex D which was then coupled with A to give Cu(III) complex E. Reductive elimination of E formed 3 and regenerated the Cu(I) catalyst. As a minor reaction pathway, intermediate B could convert to nitrogen radical F directly via a SET process and protonation. Radical coupling of A and F led to the formation of product 3 in the absence of CuCN 2\u2013H compounds and aryl azides via dual copper and visible light catalysis under redox-neutral conditions. The reaction not only intelligently utilized triplet nitrene intermediate induced by visible light, but also efficient construction of various structurally diverse benzenesulfinamide derivatives under mild conditions. The further application of nitrene intermediates in biological heterocycles synthesis is still under research in lab.In summary, we have developed a green and economical method for S(O)"} +{"text": "Since domestication, sheep and goats have provided humankind with food including meat and milk, along with wool and leather. Unlocking the genetic potential and promoting the further improvement of economically important traits is key to guaranteeing the sustainable management of sheep and goat populations in the coming decades. In this regard, the implementation of genomic tools facilitates the selection progress and provides valuable information to design educated conservation programs.Ncube et al.). Growth Hormone 1 (GH1) and Insulin Like Growth Factor 1(IGF 1) were found to be highly expressed in goats raised under intensive rather than extensive breeding systems.Goat genomics is a rapidly evolving field; however, there are several unexplored aspects in need to be unveiled to understand the genetic mechanisms underlying traits of practical interest . Genes expressed in the pituitary gland are key regulators of animal growth processes. Possible differences in the expression patterns of relevant growth-related genes obtained from the pituitary gland of 36-week-old animals have been explored in goats raised under different management systems . A reference population size of 1,500 was proposed as optimal to achieve noticeable genetic progress in genomic selection for fiber diameter and live body weight in Cashmere goats. The accuracy of the GEBV was higher based on 100 QTLs for live body weight and on 50 QTLs for fiber diameter.The effectiveness of the genomic selection depends on the applied DNA array density and optimal reference population size. Based on simulations, the medium marker density panel (45k SNPs) was found to ensure accuracy in estimating the genomic estimated breeding values (GEBV) and was recommended for genomic selection in goats . Gene ontology analysis illustrated that detected DEGs were mainly involved in testis development and spermatogenesis. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed the presence of several reproduction-associated DEGs such as collagen type I alpha 1 chain (COL1A1), collagen type I alpha 2 chain (COL1A2), Platelet Derived Growth Factor Subunit A (PDGFA), and IGF). Based on using weighted gene co-expression network analysis (WGCNA), Xu et al. reported hub genes positively associated with testis size and negatively associated with testis size. The first hub included Ran binding protein 9 (RANBP9), Dynein Axonemal Heavy Chain 17 (DNAH17), Spermatogenesis Associated 4 (SPATA4), Calcium and Integrin Binding Family Member 4 (CIB4), and Spermatid Maturation 1 (SPEM1) genes. The second one comprised of CD81 Antigen (CD81), C-Terminal Src Kinase (CSK), PDGFA, Vimentin (VIM), and Inhibin Subunit Beta A (INHBA).Male fertility including testis size is among the key factors of effective artificial insemination. Transcriptomic profiling of sheep testes at different developmental stages was explored based on the identification of differentially expressed genes (DEGs) by RNA-Seq . Case-control GWAS detected two significant SNPs within introns of the gene adhesion G protein-coupled receptor B3 (ADGRB3) associated with lower fecal egg counts. Linear regression analysis revealed four significant SNPs located within the first intron of the gene EGF-like repeats and discoidin domains 3 (EDIL3). Further, Notter et al. studied single nucleotide polymorphism effects on lamb fecal egg count (FEC) estimated breeding values in progeny-tested Katahdin sires. On chromosome 5 the largest SNP effects were at 63, 67, and 70\u00a0Mb, with LD among these SNP of r2 \u2264 0.2. Chromosome-level regional heritability mapping indicated that one 500-SNP window between 65.9 and 69.9\u00a0Mb accounted for significant variation in PFEC EBV. This window included rarely identified markers for parasite resistance as the interleukin 12B gene (IL12B) at 68.5\u00a0Mb.The safety and health of lambs is the basis for cost-effective breeding. Gastrointestinal nematodes are parasites causing suffering in young animals and significant economic losses to sheep owners. The attempt to unlock the genetic variants associated with resistance or susceptibility to this parasite was performed based on linear regression and case-control genome-wide association studies in Katahdin sheep . To achieve this goal, the authors implement a novel approach called Haplotype Richness Drop (HRiD) using only the information contained in male haplotypes. Besides three classical approaches as extreme Runs of Homozygosity islands, integrated Haplotype Score, and the number of Segregating Sites by Length were used as well. In total, 14 positive selection signals containing 34 annotated genes were identified. The most reliable selection signal was mapped by all four approaches in the region, overlapping between 13.17 and 13.60\u00a0Mb, and assigned to the CA5B, ZRSR2, AP1S2, and GRPR genes. The reported HRiD offers an interesting possibility to be used complementary to other approaches or when only males are genotyped, which is often the case in genomic breeding value estimations.Signatures of positive selection in the sheep genome are usually detected on autosomes, while no such information is available for the X-chromosome. The search for adaption-related genes was performed to provide a map of the positive selection of the X-chromosome in a sample of eight native Croatian breeds representing the East Adriatic metapopulation and local mouflons ("} +{"text": "The BioFire FilmArray\u00ae Pneumonia Panel\u00ae (PP) is a rapid diagnostic test that detects select bacterial and viral respiratory pathogens and the presence of certain antimicrobial resistance genes within 75 min of testing. Starting Nov 2021 respiratory specimens from all adult ICU patients (pts) with quantitative cultures (cx) obtained by bronchoalveolar lavage (BAL) or tracheal aspirate (TA) were automatically tested with PP. The purpose of this study was to evaluate the clinical impact of the PP.This single-center, retrospective cohort study compared patients prior to (PRE) (Jan - Mar 2021) and after (POST) (Jan - Mar 2022) implementation of the PP. Adult ICU pts with a quantitative cx obtained by BAL or TA during the study periods were reviewed in random order until 25 pts from each study month meeting study criteria were identified (75 pts in each cohort). Pts with another infection other than bacteremia with the same causative pathogen requiring antibiotics (ABX) in the previous 14 days through 5 days after specimen collection or pts who died within 5 days after specimen collection were excluded. ABX therapy for the 5 days after cx collection was evaluated. The primary outcome was time to first ABX change based on microbiologic test results. Secondary outcomes included ABX days of therapy (DOTs), time to adequate therapy, and potential vs actual ABX changes made based on the PP in POST cohort.PRE pts were a median of 54 yrs old and 56% female compared to 59 yrs and 32% female in POST cohort. Median time to first ABX change based on microbiologic data was 50.3 hrs PRE vs 21.2 hrs POST (p=0.0006). 56 (75%) of POST ABX regimens were eligible for change based on PP results; actual change occurred in 30 (54%) of these regimens . Median ABX DOTs were 8 PRE vs 6 POST (p = 0.07). For the pts with ABX changes made based on PP results, median time to first ABX change was 10 hrs. For pts who were initially on inadequate therapy, time to adequate therapy was 67 hrs PRE vs 37 hrs POST.The PP was associated with decreased time to first ABX change and fewer ABX DOTs. The PP may have had a larger impact if a higher percentage of potential ABX changes were implemented. The PP may be a useful ABX stewardship tool.All Authors: No reported disclosures."} +{"text": "T. vaginalis clinical isolates\u2019, with the sentence \u2018The actin gene of samples 3 (OP557698) and 4 (OP557697) and the T. vaginalis standard strain were selected for further sequence analysis.\u2019Following publication of the original article , the autThe authors thank you for reading this update."} +{"text": "Lianas, climbing woody plants, influence the structure and function of tropical forests. Climbing traits have evolved multiple times, including ancestral groups such as gymnosperms and pteridophytes, but the genetic basis of the liana strategy is largely unknown. Here, we use a comparative transcriptomic approach for 47 tropical plant species, including ten lianas of diverse taxonomic origins, to identify genes that are consistently expressed or downregulated only in lianas. Our comparative analysis of full-length transcripts enabled the identification of a core interactomic network common to lianas. Sets of transcripts identified from our analysis reveal features related to functional traits pertinent to leaf economics spectrum in lianas, include upregulation of genes controlling epidermal cuticular properties, cell wall remodeling, carbon concentrating mechanism, cell cycle progression, DNA repair and a large suit of downregulated transcription factors and enzymes involved in ABA-mediated stress response as well as lignin and suberin synthesis. All together, these genes are known to be significant in shaping plant morphologies through responses such as gravitropism, phyllotaxy and shade avoidance. Summary: The full-length fraction of liana transcriptomes mapped on a protein\u2013protein interactome revealed the nature of their convergence through distinct sets of expressed and downregulated genes not observed in free-standing plants. Lianas are woody vines that have evolved multiple times since at least the Devonian period in vascular plant lineages, including in the Gnetales and repeatedly in the Angiosperms, with fascinating diversity in stem anatomy even within a single genus . The NeoThe basic advantage of the liana life form is clear: lianas harvest light from the treetops without investing in the woody structures necessary to support a canopy of leaves . Several2 assimilation rates and stomatal conductance decrease while respiration increases (Vitis vinifera (grapevine from here in) we know that lianas can employ a range of anisohydric and isohydric control of stomatal conductance as water use strategy. Through their cheap but efficient leaves, lianas may also be carrying out sufficient levels of photosynthesis rapidly before midday to escape from the sun's pernicious rays.The direct solar radiation and high heat in the canopies, where most liana leaves are deployed can impose significant constraints for lianas. For instance, the photosystem II has been shown to be vulnerable to temperature and light stress in a liana as compared to its tree relative . Canopy ncreases . Seasonancreases . We, theThere has been some research on stress responses of a few liana species . HoweverFig.\u00a0S1, Dataset S1). Using network analyses, we ask the following questions: (1) what genes are uniquely expressed and downregulated in some or all of the lianas? (2) Are there unique genetic similarities among lianas related to their shared ecological challenges ? And (3) are there unique genetic similarities in lianas that reflect specific metabolic pathways underpinning the building of structural biomass?Here, we provide a comparative transcriptomic analysis across a phylogenetically diverse group of trees, shrubs, and lianas. We use reference transcriptome assemblies of 37 tree and shrub species and ten liana species . Quality metrics of transcriptomes generated by Quality Assessment Tools for Genome Assemblies (QUAST) showed the transcriptomes range in size from 49,040 to 182,060 (Ixora ferrea) (Dryad repository). The overall alignment rates for the de novo assembled reference transcriptomes, validated by Bowtie2 alignments of trimmed and quality filtered reads, were between 83 and 94% . The assembled transcriptomes captured an informative fraction of the expressed genes from 47 species where Benchmarking Universal Single-Copy Orthologs (BUSCO) analyses were able to represent between 52 and 92% of the benchmark eukaryotic genes within the Embryophyta . Annotation and filtering of non-plant transcripts through EnTAP (Eukaryotic non-model Transcriptome Annotation Pipeline) resulted in a full-length set of grapevine orthologs ranging from 6001 (Smilax coriacea) to 9488 (Dolichandra unguis-cati) for cross-comparison (Dataset S1). An ordination of key LES traits, leaf area (LA) and specific leaf area (SLA), reflected difference between lianas and non-lianas in our transcriptome set, indicating a phenotypic portrayal of liana biology at a local island scale .Using a phylogenetically diverse sample of ten liana species, we identified major convergent gene expression patterns that contrast with 37 coexisting species of trees and shrubs (Dataset S1). After the exclusion of common proteins, a protein\u2013protein interactome network based on curated STRING database was constructed using transcripts exclusively expressed by lianas and non-lianas. The total size of the network including vertices with first and second shells of interaction consisted of 3835 nodes and 14,574 edges. Grapevine has 33,568 protein-coding genes and the network occupies a little over 11% of its gene space. Close to half of the network (1687 nodes) comprised genes solely expressed by non-lianas . This protein-serine/threonine phosphatase is a key enzyme in cGMP-dependent signaling, abscisic acid perception, commitment to cell cycle and is found within the additional shells of interaction in a region of the network subtending nodes exclusively expressed in lianas . The subsequent highly connected nodes are all found within the co-downregulated portion of the network . We found no genes that are unanimously expressed by all lianas. Two nodes, shared by five lianas (L5), corresponded to the ethylene responsive transcription factor SHINE2 (SHN2) and the xyloglucan glycosyltransferase 4/cellulose synthase-like C family 4 (CSLC4) enzyme, representing the highest points of convergence via individual gene transcripts .The transcripts expressed exclusively by more than one liana (L2\u2013L5) species generated a network of 705 nodes with 1609 edges ; Fig.\u00a0S3nscripts . The nexnscripts ; Fig.\u00a0S3The combined controlled vocabulary gene ontology (GO) analysis of transcripts shared by more than two lianas (L2\u2013L5) through gProfiler displayed enriched molecular functions pertaining to the phytohormone auxin, such as auxin efflux transmembrane transporter activity (GO:0010329) and auxin transmembrane transporter activity (GO:0080161) A. BiologDataset S1).The term auxin and gravitropic response identified by gProfiler were consolidated around five genes including chaperone protein dnaJ 15-like (VIT_01S0011G03790), ABC transporter B family member 1 (VIT_08S0007G05060), protein kinase PINOID-like (VIT_10S0003G04320), auxin efflux carrier component (VIT_11S0052G00440), SEC7 domain-containing protein (VIT_02S0012G01790), and auxin efflux carrier component (VIT_14S0108G00020) (Dataset S1). Of these, 1687 had hits in the STRING interactome database .Three families belonging to Bignoniaceae, Sapindaceae, and Malpighiaceae included pairs of liana and non-liana members in our dataset. We scrutinized these with the anticipation of detecting the most informative gene expression differences . EnrichmFig.\u00a0S1, Dataset S1). At early developmental stages, transcriptomic differences between woody lianas and herbaceous vines are most likely minimal. For this reason, we included three herbaceous climbers in the dataset with the assumption that leaves of woody and herbaceous climbers would experience similar constraints independent from their stem morphology and would perform in parallel . Modifications of cell wall components and cuticle properties are beneficial for lianas in developing leaves with high SLA and optical properties since excessive light levels may damage light harvesting complexes. Reduced production cost due to altered secondary cell wall composition, increased rates of carbon fixation and fast tissue turnover in most liana species may be an outcome of series of mesophyllic adaptations. Facilitation of carbon capture and diffusion through an active carbon concentrating enzyme capable of scavenging low levels of CO2 floating inside the leaf could be adaptive since leaves with high SLA can accommodate only a few mesophyll cell layers. Lianas could be achieving more with less by increasing gene dosage through selective endoreduplication. In lianas, under hydraulic constraints, machinery evolved for the repair of damaged nuclear genetic material may be primed as early as the seedling stage.Top co-expressed genes in our analysis correspond to enzymes synthesizing or modifying cell wall components, cuticle properties, carbon concentrating mechanism, cell cycle progression through endoreduplication and double-stranded DNA damage response ; Fig.\u00a0S3Fig.\u00a0S3). SHNs together with wax inducers (WINs) stimulate biosynthesis of cuticular wax in leaf epidermis and reduce stomatal density . All 149 genes belonging to AP2/ERF protein family in grapevine genome have been transcriptionally characterized and the Arabidopsis SHN2 ortholog gene (GSVIVP00032652001/VvERF044) also showed an upregulated pattern in the grapevine leaves . A composition of esterified and de-esterified pectin residues control calcium-mediated stiffness of gel matrix within the cell wall and contributes to diversification of organ shapes through directional growth. Arabidopsis mutants show that xyloglucans and pectin can together influence shoot meristem and phyllotaxis . These cFig.\u00a0S3) . Less liFig.\u00a0S3) . RemovalFig.\u00a0S3) . OverexpFig.\u00a0S3) . This coFig.\u00a0S3) . MicrotuFig.\u00a0S3) . CelluloFig.\u00a0S3) . QuintupFig.\u00a0S3) . AnotherFig.\u00a0S3) ; Fig.\u00a0S3llotaxis . Cell wallotaxis . For insllotaxis A . The carbon concentrating mechanism describes the enzymatic pathways the atmospheric CO2 flows through the mesophyll cell wall and the plasma membrane into the liquid cytosolic phase imposing limitations on diffusion known as mesophyll conductance. Mesophyll conductance is a significant driving factor in LES . From the LES perspective, leaf construction is the sum of spatially controlled cytoplasmic growth and cell division. During leaf expansion, a plant can do more with less through a process called endoreduplication where distinct cells may increase their genomic content and gene dosage without cell division . Compatible with FBL17, some of the enriched GO terms for the top ten most connected nodes include (GO:0022402) cell cycle, (GO:0007049), double-strand break repair via break-induced replication (GO:0000727) (Dataset S1).We have observed that four lianas exclusively expressed the plant-specific FBL17 transcript that serves as a major checkpoint in cell cycle progression from Gap1 (G1) into Synthesis (S) phases Table\u00a02; Fig.\u00a0S3division . During division . In plandivision . Loss ofdivision . Liana ldivision . Significance of MYB-R2R3 co-downregulation for lianas could include stomatal control, lignification and plant architecture including phyllotaxis. The MYB-R2R3 serve as major activators and repressors controlling diverse functions such as pectin, lignin and suberin mediated xylogenesis through control of the phenolic acid metabolism, modulation of developmental signaling, cell cycle, epidermal cell fate and patterning .MYB genes are found in all eukaryotes and is the largest transcription factor family in Arabidopsis . The MYBFig.\u00a0S3). Lianas adopted features such as smaller and low-density stomatal openings with sunken guard cells for reduction of water loss due to excessive transpiration as early as the Paleozoic based on cuticular analysis of pteridosperm lianas .One particular R2R3-MYB downregulated in our analysis is MYB44 that deserves special attention since it is heavily involved with the core node of the interactome the protein phosphatase PP2C ; Fig.\u00a0S3m lianas . The sesm lianas . The behm lianas Table\u00a02Fig.\u00a0S3. m lianas . In the m lianas . Moreovem lianas . Inductim lianas . In lianm lianas . Downregm lianas . The secondary messenger cyclic GMP binds and inhibits the phosphatase activity of this PP2C in favor of kinase activity. For this reason, this PP2C is known as cGMP-dependent protein kinase (PKG) and it phosphorylates the transcription factor GAMYB to upregulate gibberellic acid-responsive genes (One noteworthy observation is that the top connected node forming the core of our interactome is another PP2C with kinase activity belonging to clade L protein phosphatases (VIT_11s0016g03430.t01/AT2G20050) . For instance, overexpressed SHN2 suppresses many MYBs involved in lignified secondary wall biosynthesis as evidenced from poplar . MYB46 targets a set of 13 genes in lignin biosynthesis . MYB48 appears to play role in xylogenesis but its mechanism of control is unclear containing evidence of alternative splicing through intron-retention . Branching of the shoots into axillary meristems is controlled by MYB37 and MYB38 also known as regulator of axillary meristems (RAX1 and RAX2), which are positively controlled by a WRKY transcription factor excessive branches 1 (EXB1) (MYB98 can be expressed in trichomes and has 83 downstream target genes in synergids . Downreg1 (EXB1) . Transcr1 (EXB1) . Their d1 (EXB1) . In our 1 (EXB1) .This study explores transcriptomic signatures related to liana leaf properties. Compared to trees, liana leaves generally carry traits colloquially conceptualized as \u2018fast and furious\u2019 type life-history strategy associated with quick growth, low cost, high turnover, high capacity for water movement, minimal investment on defense. Our results are in accordance with the LES encapsulating construction costs, rates of carbon fixation and tissue turnover. A set of uniquely expressed enzymes in charge of cell wall building, epicuticular wax synthesis, carbon capture, cell cycle appears to complement with another set of a large number of transcription factors not expressed in lianas. Through comparative transcriptomics of a diverse spectrum of families, we believe we were able to interrogate a wide set of orthologs to contribute towards understanding some of the genetic underpinnings of biology leading to the liana growth form from the leaf perspective. As further sequencing data from additional liana species with more diverse tissue types become available, the genetic basis of this fascinatingly convergent plant form will be more comprehensible.Dioscorea polygonoides (Dioscoreaceae), (HET) Heteropterys laurifolia , (PAU) Paullinia pinnata (Sapindaceae), (SEC) Securidaca virgata (Polygonaceae), (SMI) Smilax coriaceae (Smilaceae), (DOL) Dolichandra unguis-cati (Bignoniaceae). We also included four more liana species from National Center for Biotechnology Information's Sequence Read Archive (NCBI-SRA) and One Thousand PlantGenomes (1KP) dataset (Gnetum montanum (Gnetaceae) SRR5908685, (SMISIE) Smilax sieboldii (Smilacae) SRR5134200, (PASCAE) Passiflora caerulea (Passifloraceae) 1KP id:SIZE, (SCHPAR) Schlegelia parasitica (Schlegeliaceae) 1KP id:GAKQ. A full list including non-liana species can be found in the Supplementary Information . NCBI-SRA bioproject accession for the Luquillo transcriptomic set is PRJNA837288.The Luquillo transcriptomic set included six lianas (DIO) dataset with theTo analyze the transcriptomes, we chose healthy and fully developed leaves from seedlings of tree and liana species distributed between 350 and 450\u2005m in elevation from the Luquillo Experimental Forest (LEF) in the north-eastern part of Puerto Rico. For each species, approximately 5\u2005g of leaf tissue was collected and placed in a 50\u2005ml polypropylene conical tube with RNAlater . Explants were cut with a razor blade prior to being placed in the tube to allow the RNAlater to penetrate the mesophyll quickly. Samples were then frozen at \u221280\u00b0C within 2\u00a0days. Rneasy Plant Mini Kit was used for RNA extraction. RNA quantification and quality metrics were carried out using a NanoDrop 2000 spectrophotometer and an Agilent Bioanalyzer 2100 RNAseq library preparations and sequencing were performed at the Beijing Genomics Institute, Shenzen, China on Illumina Hiseq 2000 sequencer generating 100\u2005bp paired-end reads.Dataset S1). Transcriptome assemblies were annotated by EnTAP using the Diamond high performance aligner interrogating four protein databases (Dataset S3). Transcriptomes were frame selected and translated into proteins through built-in GeneMarkS-T module within EnTAP (Dataset S3). Full-length protein sequences were filtered from EnTAP results and were blasted against indexed V. vinifera proteome v29720. Top hits were selected through vsearch (Dataset S2). The resulting interactome network was annotated with colored borders and fills to highlight biologically informative nodes. Phylogenetic tree of lianas and non-lianas was constructed using the TimeTree resource compiling evolutionary divergence times derived from molecular sequence data . Trimmed fastq files were assembled by Trinity v.2.6.6 with minimum contig length 300 . Qualitytein 94) . Bacteriin EnTAP (Datasetcov 0.9) . Blast rcov 0.9) . Co-downcov 0.9) (Datasetnce data . GO termnce data . ResultsD. unguis-cati we used values from G. montanum was obtained from the China Trait Database D. polygonoides, H. laurifolia, P. pinnata, S. virgata, and S. coriacea we used unpublished data obtained from seedlings from our co-authors Samantha J. Worthy and Maria N. Uma\u00f1a. Ordination of leaf traits was done by the ClustVis webserver (Dataset S1.LA and SLA trait data came from ebserver . Trait vCo-downregulation should not be confused with gene deletions observed in the genomes of many parasitic and carnivorous plants . Absence"} +{"text": "A 12-year-old male presented to the emergency department with right eye pain after being hit with a bungee cord. His visual acuity was 20/20 in the affected eye. His pupillary exam was unremarkable without an obvious afferent pupillary defect. His intraocular pressures were 15 and 19\u00a0mmHg . Slit lamp exam of the right eye revealed a corneal epithelial defect with an associated stromal defect. Although Seidel testing initially revealed a possible small aqueous leak near the wound apex, this was not appreciated ~1 hour later. Anterior chamber depths were relatively similar between both eyes. The patient was diagnosed with a partial thickness corneal wound of the right eye and Moxifloxacin ophthalmic solution was initiated. A bandage contact lens (BCL) was also placed over the right cornea. An anterior segment optical coherence tomography (AS-OCT) revealed the tangential partial thickness laceration"} +{"text": "In many South Asian (SA) cultures, cancer is stigmatized, and family members are expected to become primary caregivers. Clinicians need to be familiar with these SA needs and values to provide culturally concordant care. The South Asian Family Approaches to Disease (SAFAD) study aims to understand cultural needs of SAs managing breast cancer. We conducted semi-structured qualitative interviews with multidisciplinary clinicians at a major academic medical center about caregiver interactions, cultural dynamics affecting clinical practice, unmet patient needs, and perceptions of culturally concordant care. Participants included physicians(8), a nurse practitioner(1), a social worker(1), and a physician assistant(1) with experience in palliative care(5), hematology/oncology(4), breast cancer(3), critical care(2), and radiation oncology(1) with one to 42 years in practice. Participants identified as Caucasian/White(6), South Asian(3), African American/Black(1), and Chinese(1). Clinicians noted the following: 1)SAs have greater family involvement in care and may defer treatment decisions to family members; 2)SAs seek clinician support with cancer management and nutrition; 3)SA emotions and hesitation around sensitive topics may result in non-disclosure; 4)SAs have diverse caregiver roles; and 5)individual SA needs cannot be generalized within the diaspora. \u201cThere\u2019s still some stigma\u2026 with breast cancer\u2026 [it] may lead a patient to not want to share their diagnosis\u2026 then set up a patient for having less support\u2026 [We can] help them feel more comfortable opening up selectively.\u201d Understanding such cultural needs is essential to cultivating trust and providing person-centered care. Interventions and resources to promote culturally concordant cancer care can target education in these areas."} +{"text": "By 2060, it is estimated that this number will increase to 13.9 million . Therefore, it is imperative to gain insight into participants\u2019 personal motivations and expectations of research to advance community participation. The Preventing Alzheimer\u2019s with Cognitive Training (PACT) study is a National Institute of Health, National Institute on Aging-funded, multi-site clinical trial examining the prevention of mild cognitive impairment and ADRD through computer-based cognitive training. Across 5 locations, data were collected from 2,360 cognitively normal participants . The current project explores individuals\u2019 motivations and expectations of cognitive training (CT) utilizing a mixed-method approach by coding qualitative open-ended questions about motivation to participate and comparing how motivational themes aligned with expectations about CT from the Expectations Assessment Scale . Six themes for participant motivation emerged: direct experience with the disease (26.9%), concern about brain health and aging (23.9%), general personal interest (17.9%), general interest in research (20.1%), referral to the study (5.8%), and altruism (5.4%). After completing the initial training session, motivation themes did not differentiate satisfaction with (p=.06) or perceived success of (p=.11) the CT program. Understanding participants\u2019 motivations can further expand and optimize recruitment and retention strategies in AD prevention research. Future research will focus on how these themes influence adherence and retention and relate to participant demographic characteristics ."} +{"text": "Non-pharmacological treatment like psychotherapy is associated with less side effects than pharmacological treatment and is often considered first-line treatment towards psychiatric disorders. The extent and variation of psychotherapy treatment offered in Danish psychiatric clinics over time has not previously been studied.To examine the nationwide use of psychotherapy treatment during 2001-2020 in individuals assigned with a psychiatric disorder diagnosis at Danish psychiatric clinics.All Danish individuals aged \u2265 3 years, who were registered with 1) a psychiatric disorder diagnosis (F10-F99) or 2) had a first psychotherapy treatment during the study period 1 January 2001 to 31 December 2020, were identified in the Danish National Patient Registry.A total of 120,916 (27 %) study participants received psychotherapy treatment during the study period, most commonly individual psychotherapy (65 %) followed by group therapy (25 %). Adults (\u226518 years) were more likely to receive therapy (34 %) than children and adolescents aged 3-17 years (15 %). The proportion of treated patients was highest among women (67 %) compared with men (33 %). The median age at first psychotherapy was 25 years (ranging from 19 to 33). 59 % of patients receiving psychotherapy had filled a psychotropic prescription within one year prior to therapy onset, particularly antidepressants (44 %) and antipsychotics (22 %).The use of psychotherapy for treatment of psychiatric disorders is limited among Danish patients, although national clinical guidelines recommend it as first-line treatment of common conditions such as depressive, anxiety and obsessive-compulsive disorders.No significant relationships."} +{"text": "To characterize how antibiotic exposure impacts development and durability of intestinal dysbiosis and the acquisition of antibiotic resistance genes (ARGs) in the intestinal flora of premature infants in the Neonatal Intensive Care Unit (NICU), we established an infant gut microbiome biorepository (IGMB).We performed prospective weekly stool collection in NICU patients meeting the following criteria: birthweight < 2000 g, postnatal age < 2 months and no diagnoses of congenital gut malformation or cyanotic heart disease. Cases were infants with bloodstream infections (BSI), defined as bacterial growth from blood culture; controls were infants with < 5 days of antibiotic exposure, no BSI nor necrotizing enterocolitis. We performed metagenomic analysis on 5\u20136 serial stool samples from each of the 10 cases and 10 controls (n= 100 stools). We used Wilcoxon rank sum tests for pairwise comparisons.Enterococcus spp. . Controls did not have similar trends in Enterococcus spp., but did have higher relative abundance of facultative anaerobes including Bifidobacterium, Lactobacillus, and Veillonella spp. . Antibiotic resistance gene (ARG) abundance did not differ significantly between cases and controls.From July 2021 to May 2022, 265 infants contributed 1,300 stool samples to the IGMB. In 7 of 8 BSI cases the causative pathogen was identified in the pre-BSI stool sample. Two more BSI cases did not have a pre-BSI stool sample. Microbiome species \u03b1-diversity increased with advancing postnatal age in controls but not in cases . Among 6 cases with high beta lactam exposure ( >14 cumulative days by last stool collection), 3 had notable increases in the relative abundance of Enterococcus spp. prevalence with higher beta lactam exposure. ARG abundance did not differ between cases and controls. The clinical implications of this microbiome dysbiosis warrant further study.Compared to controls, neonates with BSI have increased antibiotic intensity throughout their NICU admissions, reduction of microbial species diversity and overabundance of specific organisms in their gut microbiomes. We observed increased J\u00f6rn-Hendrik Weitkamp, MD, Roche Diagnostics: Advisor/Consultant."} +{"text": "This audit covered 3 hospitals in Glasgow City which has 1221 beds providing inpatient healthcare for the north east region of the city. To improve the referral process,we aimed to verify adherence to existing referral pathway and adequacy of information provided by referrals. Referral characteristics including referral indication, intervention and outcomes were accounted for to identify area interest that may help improve the referral process.Our referral pathway involves completion of a Microsoft Word referral template subsequently sent electronically to an internal electronic mail.Referrals in a 2 month period were included in the audit. Each referral was reviewed for adherence to the referral template, adequacy of provided information and referral indications. Intervention in the form of staff input, Mental Health Act status, psychotropic medication prescribed and given diagnosis was ascertained via staff electronic entry records.139 referrals were included. 114 referrals (82%) adhered to the referral template. 72 referrals (52%) contained adequate information. Common referral indications were delirium (23%), agitation (20%), low mood (18%) and cognitive decline queries (18%). Staff input ranged from psychiatrist input (46%), liaison nurses (40%), clinical psychology (1%) and shared input (13%). 16 referrals (12%) resulted in subsequent detention under the Mental Health Act. Psychotropic medications prior to liaison assessment included antidepressants (49%), antipsychotics (29%) and benzodiazepines (16%). Liaison assessment resulted in increase use of antipsychotic (55%) and reduction of antidepressants (29%) and benzodiazepines (10%), Delirium (34%), dementia (21%), Mood & Anxiety related disorders (18%) and Query of Cognitive Impairment (14%) were recorded as the most discussed diagnosis.Referrals with inadequate details affect the service's ability to efficiently assess for clinical urgency and matching of appropriate interventions to suit clinical needs. The percentage difference in delirium between referral indication and diagnosis highlights that delirium can be under-recognised, resulting in potentially delayed treatment. Identifying common given diagnosis and differences in psychotropic medication prescribing pattern points to the need for training and support of acute medical ward staff in utilising therapeutics for management of acute mental health disorder.A pending electronic referral pathway with mandatory entries and linked relevant online resources can encourage early recognition of acute mental health disorder and prompt early management including the use of appropriate therapeutics. An additional feature allowing direct referrals by acute ward staff to community mental health team would support continuity of care for discharged patients needing ongoing mental health assessment."} +{"text": "We read with great interest the research article entitled \u201cEffect of the location and size of thyroid nodules on the diagnostic performance of ultrasound elastography: A retrospective analysis\u201d by Xie and Yu . The autAccording to the Bethesda System for Reporting Thyroid Cytopathology (TBSRTC), 1st ed., the risk of malignancy (ROM) from indeterminate cytology is as follows: III, atypia of undetermined significance or follicular lesion of undetermined significance (AUS/FLUS), 5%\u221215%; IV, follicular neoplasm (FN) or suspicious for a follicular neoplasm (SFN), 15%\u221230%; and V, suspicious for malignancy (SM), 60%\u221275%, while in the TBSRTC 2nd ed., the ROMs are 10%\u221230%, 25%\u221240%, and 50%\u221275%, respectively.The leading thyroid cytology classification systems have categorized indeterminate cytology and their associated ROMs into categories equivalent to III, IV, and V of the TBSRTC.The current 2016 UK Royal College of Pathologists (RCPath) Thy terminology divides the indeterminate cytology category Thy3 into Thy3a, neoplasm possible, atypia/nondiagnostic, and Thy3f, neoplasm possible, suggestive of FN, with ROMs of 20%\u221231%, and 24%\u221239%, respectively, and defines Thy4 as SM with a ROM of 70%\u221287%.The 2014 Italian Consensus for the Classification and Reporting of Thyroid Cytology (ICCRTC) divided its diagnostic category TIR3, indeterminate cytology, into two subcategories: TIR3A and TIR3B , with different expected cancer risks and discrete clinical characteristics. TIR3A is characterized by augmented cellularity with numerous microfollicular structures in a low colloid background or a scarce cellular structure consisting predominantly of microfollicular groups with oxyphilic features (Hurthle cells), with an expected cancer risk of <10%. TIR3B is characterized by mild/focal nuclear atypia, with an expected cancer risk of 20%\u221230%. The ICCRTC also includes category TIR4, SM with an expected malignancy risk of 60%\u221280%.The 2014 Royal College of Pathologists of Australasia (RCPA) and Australian Society of Cytology (ASC) categorized thyroid nodules as follows: 3, indeterminate (AUS/FLUS) with a low ROM of 5%\u221213%; 4, suggestive of an FN (FN/SFN) with a moderate ROM of 21%\u221226%; and 5, SM with a high ROM of 85%\u221290%.The 2013 Japan Thyroid Association (JTA) Guidelines for the Management of Thyroid Nodules categories are as follows: indeterminate B, other tumors, (TBSTRC III); indeterminate A, FN, (TBSRTC IV), i) indeterminate A1, favor benign, ii) indeterminate A2, borderline, iii) indeterminate A3, favor malignant; and SM (TBSRTC V).In addition, the first one remains a crucial challenging cytological category in all systems. Category I in TBSRTC, 1st and 2nd ed., is Nondiagnostic or Unsatisfactory. Category Thy1 in the 2016 RCPath is Nondiagnostic for cytological diagnosis and Thy1c is Nondiagnostic for cytological diagnosis-cystic lesion. Category TIR1 in the 2014 ICCRTC/Italian Society for Anatomic Pathology and Cytology\u2212Italian Thyroid Association (SIAPEC-AIT2013) is Nondiagnostic and TIR1c is Nondiagnostic cystic. Category 1 in the 2014 RCPA/ASC is Nondiagnostic, and Category 1 in the 2013 JTA is Inadequate. These categories are still used in the management of suspicious (not just indeterminate cytology) nodules 2. Therefo"} +{"text": "A 37-year-old male patient presented with 1-month history of pain over the bulb of penis during retraction of foreskin. Patient suffered from type 1 diabetes mellitus on poor glycemic control. On examination multiple white patches of 1 mm x 3 mm dimension were observed with pain during retraction of prepuce. Smegma deposition over the glans penis and erythematous areas were revealed while scraping the lesions. The patient, screened for urinary tract infection (UTI) and sexually transmitted disease (STD) including hepatitis B, syphilis and HIV which were negative and complete blood count was normal. Since smegma can be a precursor for genital infections, physicians must scrupulously examine diabetic patients presenting as timely diagnosis and treatment would improve patient\u00b4s quality of life. The patient, put on long acting insulin and advised personal hygiene and showed significant improvement during his follow-up visit, 1 month later."} +{"text": "Leishmania persist in a high proportion of individuals despite clinical resolution, indicating that determinants other than parasite clearance are involved in drug efficacy.Control of cutaneous leishmaniasis (CL) relies on chemotherapy, yet gaps in our understanding of the determinants of therapeutic outcome impede optimization of antileishmanial drug regimens. Pharmacodynamic (PD) parameters of antimicrobials are based on the relationship between drug concentrations/exposure and microbial kill. However, viable h1 and Th17 gene signatures were characterized in peripheral blood mononuclear cells (PBMCs) during treatment with meglumine antimoniate (MA) and clinical cure of human CL caused by Leishmania (Viannia). We explored relationships of immune gene expression with plasma and intracellular antimony (Sb) concentrations.In this study, the profiles of expression of neutrophils, monocytes, TOur findings show a rapid and orchestrated modulation of gene expression networks upon exposure to MA. We report nonlinear pharmacokinetic/pharmacodynamic (PK/PD) relationships of Sb and gene expression dynamics in PBMCs , concurring with a time lag in the detection of intracellular drug concentrations and with PK evidence of intracellular Sb accumulation.Our results quantitatively portray the immune dynamics of therapeutic healing, and provide the knowledge base for optimization of antimonial drug treatments, guiding the selection and/or design of targeted drug delivery systems and strategies for targeted immunomodulation. We describe the nonlinear relationship between meglumine antimoniate pharmacokinetics and immune gene expression profiles during treatment of patients with cutaneous leishmaniasis and reveal immune gene signatures as pharmacodynamic end points of antileishmanial drugs. Leishmania ) were detected throughout treatment in samples collected 1 hour after dosing (i]) were also measured within PBMCs. [Sbi] measured 1 hour after dose on the first day of treatment was under the LLOQ but measurable thereafter in samples collected 1 hour after dosing on days 10 and 20 (EoT). [Sbi] at days 60 and 90 were under the LLOQ . Intracellular time to reach Cmax (Tmax) was 3.4 hours and was 5 times lower than plasma Cmax (intracellular Cmax = 6625 \u03bcg/L \u00b1 1186 \u03bcg/L). The trough [Sbi] (C24i) was 2739 \u03bcg/L (\u00b1 497 \u03bcg/L), which was 10 times higher than plasma C24 (234 \u03bcg/L \u00b1 50.3 \u03bcg/L). The slower time-dependent decrease in [Sbi] compared with [Sbp] provides evidence of intracellular drug accumulation.Concentration-time curves and PK parameters were derived from [Sbd at EoT . Plasma Leishmania. We asked whether the lower [Sbi] could be an artifact of specific accumulation within monocytes. PBMCs from healthy donors were obtained, and monocytes were isolated using CD14+ magnetic bead sorting. Ten million PBMCs and 10 million monocytes were incubated with MA for 1 hour at plasma Cmax (32 000 \u03bcg-Sb/L). Similar [Sbi] were found in PBMCs and in isolated monocytes, indicating that Sb is incorporated within monocytes as well as other mononuclear white blood cells.The antileishmanial effect of drugs is typically measured in monocytes/macrophages as these are the preferential host cells for Leishmania (V.) panamensis .Intracellular Sb concentrations were also plotted against time-dependent gene expression profiles; cluster A genes were linearly and inversely correlated with intracellular drug concentrations (50), and derived PK/PD indices cannot completely explain complex interactions like those of intracellular pathogens that cause chronic and persistent infections and [Sbi] . With thntration , 46. Thip]. The main assumption under the collapse of hysteresis loops is that the measured effect depends on drug concentrations in the effect compartment rather than the central compartment and cluster A/E gene expression data showed an inverse linear correlation, collapsing the hysteresis loops that represented the relationships with and [Sbi] indicates nonequilibrium between the central and the effect compartment [Sb]. (2) SbV is reduced to SbIII for direct antileishmanial effect, and this can occur in mammalian as well as Leishmania cells compared with [Sbp], provides new bases for optimization of antimonial drugs through improved drug delivery systems (DDSs). To reduce toxicity and length of antileishmanial drug regimens, DDSs have been empirically selected assuming that their main benefit arises from enhanced drug concentration within infected macrophages at different anatomical sites [Modulation of systemic immune gene expression profiles throughout the course of antileishmanial treatment, together with evidence of intracellular Sb accumulation despite lower [Sbal sites . Howeveral sites . The datClinical Infectious Diseases online. Consisting of data provided by the authors to benefit the reader, the posted materials are not copyedited and are the sole responsibility of the authors, so questions or comments should be addressed to the corresponding author.Supplementary materials are available at ciaa1206_suppl_Supplementary_MaterialClick here for additional data file."} +{"text": "Safety net HIV providers face operational challenges during the COVID pandemic with services often transformed to telehealth. HIV infected persons are a priority population for SARS CoV-2 vaccination. Medical mistrust of COVID vaccines has been cited as a contributor to vaccine hesitancy. Data on efficient and successful vaccination efforts of HIV infected persons in safety net health systems is needed. In San Mateo County, Latino persons comprised 42% of all COVID cases, Whites 16%, and African Americans 2%.SARS CoV2 vaccination with BNT162b2 (Pfizer\u2013BioNTech), mRNA-1273 (Moderna) or Ad26.COV2.S (Janssen) vaccine were offered beginning February 2, 2021 through May 28, 2021 in a northern California public County HIV clinic. Clinic patients were contacted by bilingual English/Spanish speaking HIV clinic staff and appointments scheduled at County affiliated vaccination sites. Clinic staff followed up by phone with patients who did not initially accept vaccine. We calculate the percentage of patients who completed vaccine series and use multivariable logistic regression analysis to estimate the odds of series completion by patient race/ethnicity, gender and age.Virtually all, 95% (349/365) of HIV patients in our County HIV clinic were offered vaccine during a 17 week period. Among those, 86% (313/365) accepted and received at least one dose and 80% completed the series (292/365) at time of this analysis. Janssen vaccine was given to only 2% (7/313) patients. Series completion was highest among Latinos and Asians. Latinos had the highest odds of vaccine series completion . COVID-19 Vaccine Series Completion in a California Public HIV Clinic by Race/Ethnicity, Age and Sexual Orientation, n=364HIV patients offered SARS CoV2 vaccine by County HIV clinic staff with established patient care relationships had high vaccine acceptance (80%), comparable to 68% series completion in the county overall and 56% in the health equity quartile county census tracts. Latino HIV infected persons were most likely to complete the COVID vaccine series. Ryan White funded HIV clinics are ideal hubs to coordinate HIV patient COVID vaccination efforts. Adding COVID vaccine completion to HIV clinic performance measures would likely be beneficial.All Authors: No reported disclosures"} +{"text": "Limited evidence exists to guide the management of patients with liver metastases from squamous cell carcinoma (SCC). The aim of this retrospective multicentre cohort study was to describe patterns of disease recurrence after liver resection/ablation for SCC liver metastases and factors associated with recurrence-free survival (RFS) and overall survival (OS).Members of the European\u2013African Hepato-Pancreato-Biliary Association were invited to include all consecutive patients undergoing liver resection/ablation for SCC liver metastases between 2002 and 2019. Patient, tumour and perioperative characteristics were analysed with regard to RFS and OS.P\u2009=\u20090.134). A positive resection margin significantly influenced RFS for both anal cancer and non-anal cancer liver metastases . Median survival duration and 5-year OS rate among patients with anal cancer and non-anal cancer were 50\u2009months and 45 per cent and 21\u2009months and 25 per cent, respectively. For the entire cohort, only non-radical resection was associated with worse overall survival .Among the 102 patients included from 24 European centres, 56 patients had anal cancer, and 46 patients had SCC from other origin. RFS in patients with anal cancer and non-anal cancer was 16 and 9\u2009months, respectively . The present study shows that hepatic resection/ablation of liver metastases from SCC can result in long-term survival. There is limited evidence to guide the management of patients with liver metastases from squamous cell carcinoma (SCC). In metastatic SCC, spread to the liver seems only possible through systemic haematogenous dissemination, which explains the historical concern that hepatectomy is less beneficial for these patients. During the past two decades, several publications have reported on liver resection for non-colorectal non-neuroendocrine liver metastases, unanimously concluding that it is a safe treatment option with the potential of long-term survivalThe aim of this retrospective multicentre study was to describe patterns of disease recurrence after liver resection/ablation of SCC liver metastases and factors associated with recurrence-free survival (RFS) and overall survival (OS) through collaboration within the European\u2013African Hepato-Pancreato-Biliary Association (E-AHPBA).Members of the E-AHPBA were invited to include retrospectively all consecutive patients submitted to liver resection and/or ablation of histologically proven SCC liver metastases between 2002 and 2019. The study protocol was approved by the regional ethics board in Stockholm, Sweden (Dnr 2019\u201304681). All participating centres obtained ethical approval according to national/local legislation before inclusion of patients. Participating centres entered all data into a web-based application for collecting data, Research Electronic Data Capture , containing predefined case report forms designed specifically for this project to collect relevant data.The following data were recorded for each patient: age, sex, ASA physical status, Eastern Cooperative Oncology Group (ECOG) performance status and Charlson co-morbidity index2 test or Fisher\u2019s exact test, the latter if sample sizes were small (10 patients or fewer). RFS was measured from date of liver resection/ablation until date of first recurrence, detected on any radiography, or death. Patients with simultaneously diagnosed extrahepatic metastases that were not treated with curative intent were excluded from the analysis of RFS and factors influencing RFS. OS was measured from date of liver resection/ablation until date of death or date of last follow-up. Survival probabilities were illustrated using Kaplan\u2013Meier graphs and the log rank test for testing equality of survival functions between groups. Patient and tumour factors potentially influencing RFS and OS were analysed using Cox proportional hazard regression model and included in the multivariable analysis if P\u2009<\u20090.200 in the univariable analysis, and reported as hazard ratio with associated 95 per cent confidence intervals. For the purposes of data analysis in the regression models, values were dichotomized based on median values. Statistical significance was set at a two-sided alpha level of 0.050. All statistical analyses were performed in STATA 15.0 .Descriptive statistics were used to depict the study cohort. Continuous variables were described as median with range and differences tested with Wilcoxon rank-sum test. Categorical variables were specified with frequencies (percentage) and differences in proportions were analysed with Pearson\u2019s \u03c7Some 102 patients were included from 24 European institutions; 80 of these patients were diagnosed and treated for liver metastases after 2010. There were 56 patients with anal cancer liver metastases and 46 patients with non-anal SCC liver metastases. The baseline patient and tumour characteristics are summarized in Table 2. A higher proportion of patients with synchronous liver metastases and multiple liver metastases were treated with systemic chemotherapy while primary tumour origin , extrahepatic metastases (P\u2009=\u20090.811) and nodal status of primary (P\u2009=\u20090.524) did not influence chemotherapy strategy. Response to chemotherapy was reported for 37 of 61 patients: nine patients had stable disease, 24 had partial response and four patients had tumour progression.Chemotherapy was administered to 61 out of 102 patients; details on regimen strategy and other perioperative characteristics for the entire cohort are summarized in Treatment with curative intent of simultaneously detected extrahepatic metastases was given to 11 of 16 patients. The five patients whose extrahepatic metastases eventually were not treated with curative intent originated from anal cancer (1 patient), cervical cancer (2 patients) and gallbladder cancer (2 patients).Table 2). Median length of hospital stay was 7 (range 0\u201387) days). Four patients died within 90\u2009days of surgery of which one death was a direct consequence of the surgical interventions (Clavien\u2013Dindo V).Liver resection only was carried out in 87 patients, 10 patients had a combination of resection and thermal ablation and five patients underwent thermal ablation only. Median operative time was 196 (range 40\u2013580) minutes). Major complications occurred in nine patients within 30\u2009days of surgery (During a median follow-up time of 22 (range 0.5\u2013145) months from liver resection/ablation, recurrent liver metastases in the entire cohort were diagnosed in 42 patients. The metastases were solely intrahepatic in 11 patients and intrahepatic metastasis was associated with extrahepatic metastasis in 30 patients. Of those with any liver recurrence (42 patients), 15 patients had a second hepatectomy. A further 21 patients suffered from extrahepatic recurrence only, including recurrence at primary tumour site, and none of them had surgery for these extrahepatic metastases or re-resection of primary site recurrences. The extrahepatic recurrences (51 patients) were most often first diagnosed in the lung (24 patients), followed by distant lymph node metastases (2 patients), peritoneum (6 patients), bone (5 patients), brain (3 patients) and other (8 patients); extrahepatic recurrences were in a single organ in 31 of 51 patients and were diagnosed at multiple locations in the remaining 20 patients.In the subgroup of patients with anal cancer liver metastases (56 patients), 22 had intrahepatic recurrence after liver resection, five had simultaneously diagnosed extrahepatic recurrence and a further 11 patients later developed extrahepatic recurrence. Of these 22 patients, 11 had a repeat liver resection of which nine were considered to have curative intent.Table 3. In patients resected for liver-metastatic anal cancer, the median RFS was 16 (95 per cent c.i. 8 to not reached) months and in the non-anal cancer group the median RFS was 9 (95 per cent c.i. 7 to 18) months, log rank test P\u2009=\u20090.134 . The only factor significantly influencing RFS among patients resected for liver-metastatic anal cancer was a positive resection margin (Table 4). Factors independently influencing RFS negatively in the entire cohort (93 patients) were an R1 resection and non-anal primary .The median RFS after liver resection in the entire cohort (excluding five patients whose simultaneous extrahepatic metastases were not treated curatively and four patients with missing data on recurrence) was 11 (95 per cent c.i. 8 to 19) months, as outlined in Table 3). A favourable survival was seen for liver-metastatic anal cancer with a median duration of survival and 5-year OS rate of 50 (range 30 to not reached) months and 46 per cent respectively, compared with 21 (range 14\u201346) months and 25 per cent respectively, in liver-metastatic non-anal cancer, P\u2009=\u20090.006 . The median duration of survival and 5-year OS for men resected for anal cancer liver metastases was 27\u2009months and 25 per cent respectively, and the corresponding survival data for women were 61\u2009months and 53 per cent (P\u2009=\u20090.023). On multivariable analysis on factors influencing survival among anal cancer liver metastases, no individual factor reached significance (Table 4). The only factor remaining significantly prognostic of worse OS in the entire cohort was a positive resection margin . Excluding the ablated patients from the regression analysis did not alter the result regarding the significant impact of resection margin on RFS or OS . Median duration of survival and OS rate for different primary tumours origin of non-anal SCC are outlined in Table 3.At the end of follow-up, 51 patients were still alive, resulting in a median OS of 37 (95 per cent c.i. 26 to 50) months, with corresponding 1-, 3-, and 5-year OS of 85, 51 and 36 per cent respectively, for the entire cohort .Disclosure. The authors declare no conflict of interest."} +{"text": "Scientific Reports 10.1038/s41598-020-78571-0, published online 11 December 2020Correction to: This Article contains a typographical error in the first paragraph of the Results section, where\u201cThe data of 89 participants are presented, of which 39 had tinnitus and hearing loss (THL group), 21 participants had hearing loss but no tinnitus (HL group), and 39 controls with neither tinnitus and no or minimal hearing loss (CO group).\u201dshould read:\u201cThe data of 99 participants are presented, of which 39 had tinnitus and hearing loss (THL group), 21 participants had hearing loss but no tinnitus (HL group), and 39 controls with neither tinnitus and no or minimal hearing loss (CO group).\u201d"} +{"text": "Funding and additional information should read as follows:10.13039/100007553Aids Fonds Netherlands Grants 2005021 and 2008013, the 10.13039/100000865Bill and Melinda Gates Foundation Collaboration for AIDS Vaccine Discovery (CAVD) Grants OPP1111923 and OPP1132237, 10.13039/100000002National Institutes of Health Grant P01 AI110657 and funding from 10.13039/100010661European Union\u2019s Horizon 2020 research and innovation programme under grant agreement No 681137.This work was supported by"} +{"text": "Radix species. Then, we estimated the ancestral distribution using the geographic coordinates and colonization routes. In addition, a statistical test of the colonization distances in the latitudinal and longitudinal directions was performed. We also conducted ecological niche modeling for two widely distributed species using climatic data. Ancestral geographic reconstruction estimated the origin of the genus to be around the Indian subcontinental region and showed that latitudinal immigration distances were shorter than longitudinal immigration distances in the diversification process. Ecological niche models suggested that the current distribution was restricted by climate, with annual mean temperature and precipitation of the driest month as particularly strong factors. Niche conservatism to the climate can affect the diversification of freshwater snails.To clarify the effect of niche conservatism on evolutionary history, we focused on freshwater snails, which have different ecological and phylogenetic properties from previously tested taxa. We conducted a phylogenetic analysis using 750\u00a0lymnaeid individuals from 357\u00a0sites of eleven Radix.To clarify the effect of niche conservatism on evolutionary history, we conducted a comprehensive phylogeography and estimation of dispersal routes. As a results, niche conservatism can affect the diversification of freshwater snails, genus In particular, niche conservatism (NC), namely the retention of niche\u2010related ecological traits through evolutionary history and has a wide distribution range covering most of the Old World, from the tropics to the subarctic regions dispersal for each distribution transition event by reconstructing the historical distribution change under specific latitudes and longitudes. The distribution transitions of each lineage have previously been evaluated as either categorical data or indicator values according to the manufacturer's instructions. We sequenced fragments of the mitochondrial cytochrome c oxidase subunit 1 (CO1), internal transcribed spacer 2 ITS2), 28S ribosomal RNA (28S), and Histone 3 (H3) Table . The pol, 28S rib2.2Radix . Eight species were selected as outgroups from the eight most closely related genera with reference to a previous phylogenetic study and combined (CO1+H3+ITS2+28S) phylogenies to assess phylogenetic positions among each species estimated by CO1 phylogeny. We used 81 individuals from 9 ly Table . Racesin2.3To trace the historical changes in the distribution of taxa, we performed Bayesian ancestral distribution reconstruction using BayesTraits v.3.0.1 under a geographical model Pagel, , 2017. A2.4Radix distribution, we conducted ecological niche modeling (ENM) using MaxEnt version 3.4.1 monophyletic, and Radix sp. was relatively supported (BPP\u00a0=\u00a00.86/BV\u00a0=\u00a0100) monophyletic. The monophylies of R.\u00a0plicatula and R.\u00a0rufescens were not well supported but can be recognized as paraphyletic species. In addition, the topologies of both the combined tree and nuclear tree were similar to those of the CO1 tree, except for the low\u2010supported branches .Our geographic reconstruction estimated the ancestral locations of each node Table . The resE Figure . FurtherE Figure . The cal3.3R.\u00a0plicatula had an average test AUC value of 0.9742 . The ecological niche model of s Figure .3. The ts Figure .3. The o4Radix species was located around the Indian subcontinent . Furthermore, our phylogeny showed distinctive subclades within some species , the distributions of these species were concentrated around the origin of the genus and indirect adaptation (interaction with other organisms adapted to the temperature) can be considered may have determined the distribution transitions in the evolutionary history of Radix. Many phylogeographical studies have shown that distribution transitions over climate zones are difficult ; data curation (lead); formal analysis (lead); funding acquisition ; investigation (lead); methodology (lead); resources (lead); software (lead); validation (lead); visualization (lead); writing \u2013 original draft (lead); writing \u2013 review & editing (lead). Takahiro Hirano: Data curation (supporting); investigation ; resources (supporting); supervision (supporting); validation ; writing \u2013 review & editing . Bin Ye: Data curation (supporting); investigation (supporting); resources ; writing \u2013 review & editing . Larisa Prozorova: Data curation (supporting); investigation (supporting); project administration (supporting); resources ; writing \u2013 review & editing . Mohammad Shariar Shovon: Investigation (supporting); project administration (supporting); resources ; writing \u2013 review & editing . Tu Van Do: Investigation (supporting); project administration (supporting); resources ; writing \u2013 review & editing . Kazuki Kimura: Data curation (supporting); funding acquisition (supporting); investigation (supporting); project administration (supporting); resources ; writing \u2013 review & editing . Purevdorj Surenkhorloo: Project administration (supporting); resources ; writing \u2013 review & editing . Yuichi Kameda: Data curation (supporting); investigation (supporting); resources (supporting); supervision (supporting); writing \u2013 review & editing . Yuta Morii: Data curation (supporting); investigation (supporting); resources (supporting); writing \u2013 review & editing . Hiroshi Fukuda: Data curation (supporting); investigation (supporting); resources (supporting); supervision (supporting); writing \u2013 review & editing . Satoshi Chiba: Conceptualization (supporting); funding acquisition ; methodology (supporting); project administration (lead); supervision (lead); writing \u2013 review & editing .Table S1\u2010S3Click here for additional data file.Fig S4Click here for additional data file."} +{"text": "In patients with gastric cancer, 6\u201327% of patients are diagnosed with T4b disease that invades adjacent organs, and curative resection can improve the prognosis of these patients.2). Upper gastrointestinal endoscopy performed after the administration of six courses of the S-1 and oxaliplatin regimen revealed a persistent primary lower gastric wall lesion; however, the diameter of the abdominal wall invasion and metastatic lymph nodes was significantly reduced, in addition to decreased serum carcinoembryonic antigen and carbohydrate antigen 19-9 levels. Subsequently, the patient underwent distal gastrectomy with D2 lymphadenectomy combined with transverse colon and abdominal wall resection. We performed radical en bloc resection and achieved a tumor-free resection margin. Simple abdominal wall closure was performed without mesh or musculocutaneous flap placement. Histopathological examination of the resected tumor specimen showed direct invasion of the mesocolon and rectus abdominis muscle. The patient was postoperatively diagnosed with L Gre-Ant type5 T4b (SI: rectus abdominis muscle) N2 PM0 DM0 Stage IIIA R0 Grade 2a gastric cancer based on histopathological findings and received S-1 as adjuvant chemotherapy, 2 months postoperatively. No recurrence was detected 6 months postoperatively.A 70-year-old Japanese man presented with an abdominal tumor and was diagnosed with advanced gastric cancer with extensive abdominal wall invasion. We performed open gastrojejunal bypass for gastric obstruction and initiated a chemotherapeutic regimen comprising S-1 (120 mg/day) and oxaliplatin (100 mg/mWe report a case of advanced gastric cancer with extensive abdominal wall invasion that was successfully treated with gastrectomy combined with resection of adjacent organs showing tumor invasion after effective systemic chemotherapy. A therapeutic approach comprising curative surgery combined with perioperative chemotherapy is useful in patients with T4b gastric cancer. Gastric cancer (GC) is the fifth most common malignancy and the third most common cause of cancer mortality worldwide . AlthougWe present a case of advanced GC with extensive abdominal wall invasion that was successfully treated with gastrectomy combined with adjacent organ resection after effective systemic chemotherapy.2, every 21 days). Although neutropenia occurred as an adverse event associated with SOX therapy, the diameter of the abdominal wall invasion and metastatic lymph nodes was reduced after the administration of four courses of the SOX regimen. Therefore, we administered six courses of the SOX regimen. Upper gastrointestinal endoscopy revealed a persistent primary lesion of the lower gastric wall presented with an abdominal tumor and suspected advanced GC. He had previously undergone an operation for ileus, and his family history was not significant. A contrast-enhanced computed tomography scan (eCT) revealed extensive abdominal wall invasion by the main gastric tumor in addition to enlarged peritumoral lymph nodes; however, no lung and liver metastases were detected Fig. a. BiopsyCurative resection is the mainstay of treatment for patients with T4b GC \u20139. HowevIn conclusion, we report a case of advanced GC with extensive abdominal wall invasion that was successfully treated with gastrectomy combined with resection of adjacent organs showing tumor invasion after effective systemic chemotherapy. In our view, a therapeutic approach comprising curative surgery combined with perioperative chemotherapy is useful in patients with T4b GC."} +{"text": "ABSTRACT IMPACT: This work assesses clinical implementation of a surface guided imaging system to improve the accuracy radiation delivery for treatment of brain lesions using a patient CT derived head phantom. OBJECTIVES/GOALS: Advancements in radiotherapy design have made clinical demand for efficient and accurate methods to deliver stereotactic radiosurgery (SRS) for treatment of intracranial lesions. This study assesses the potential of using surface guided imaging for setup using a 3D patient specific head phantom. METHODS/STUDY POPULATION: A single isocenter, multiple metastases SRS plan was generated on a CT derived RTsafe Prime phantom made of tissue equivalent materials and a polymer gel insert. Five targets of varying diameters were treated with 8Gy of radiation using two different positioning techniques. The first gel insert was irradiated within the phantom according to internal alignment with standard orthogonal x-ray imaging while the second setup used surface guided imaging, based on external anatomy. 42 hours after irradiation, the phantom was scanned in a head coil using a 1.5T MRI. MR images were fused with the patient CT data and structure set to further evaluate calculated and measured dose distributions. RESULTS/ANTICIPATED RESULTS: Discrepancies in phantom setup according to standard orthogonal x-ray imaging compared to surface guided imaging demonstrated to be <1mm in each translational and angular directions. The 3D gel inserts permitted spatial analysis to compare dose distributions of measured values to those calculated in a treatment planning system (TPS). 3D GI (Gamma Index) analysis showed good alignment in high dose regions and resulted in passing rates >94% (5%/2mm) and >87% (3%/2mm). Finally, 3 of 5 targets showed better 3D GI passing rates and less geometric offset for positioning with the surface guided imaging. DISCUSSION/SIGNIFICANCE OF FINDINGS: 3D spatial analysis of human like phantoms demonstrated that patient positioning according to external anatomy performed comparable to standard methods aligning to the internal anatomy, for a multiple met SRS treatment."} +{"text": "Scientific Reports 10.1038/srep20320 published online 03 February 2016Correction to: This Article contains errors.In Figure 3H, the figures for the control group (BDNF+/+ mice) were mistakenly selected from the candidate representative images for IBS-D FSN group (BDNF+/+ mice).The correct Figure\u00a0These changes do not affect the conclusions of the article."} +{"text": "The novel 1PGE-THIVC Env trimer displayed a native like pre-fusion closed conformation in solution as determined by small angle X-ray scattering (SAXS) and negative stain electron microscopy (EM). This closed spike conformation of 1PGE-THIVC Env trimers was correlated with weak or undetectable binding of non-neutralizing monoclonal antibodies (mAbs) compared to neutralizing mAbs. Furthermore, 1PGE-THIVC SOSIP induced potent neutralizing antibodies in rabbits to autologous virus variants. The autologous neutralizing antibody specificity induced in rabbits by 1PGE-THIVC was mapped to the C3/V4 region (T362/P401) of viral Env. This observation agreed with electron microscopy polyclonal epitope mapping (EMPEM) of the Env trimer complexed with IgG Fab prepared from the immunized rabbit sera. Our study demonstrated neutralization of sequence matched and unmatched autologous viruses by serum antibodies induced in rabbits by 1PGE-THIVC and also highlighted a comparable specificity for the 1PGE-THIVC SOSIP trimer with that seen with polyclonal antibodies elicited in the elite neutralizer by negative-stain electron microscopy polyclonal epitope (ns-EMPEM) mapping.Evaluating the structure-function relationship of viral envelope (Env) evolution and the development of broadly cross-neutralizing antibodies (bnAbs) in natural infection can inform rational immunogen design. In the present study, we examined the magnitude and specificity of autologous neutralizing antibodies induced in rabbits by a novel HIV-1 clade C Env protein (1PGE-THIVC) env isolated from the broadly cross neutralizing plasma of an elite neutralizer. This novel SOSIP Env trimer demonstrated comparable antigenic, structural and immunogenic properties that favoured several ongoing subunit vaccine design efforts. Interestingly, ns-EMPEM analysis suggested the novel clade C SOSIP induced a comparable neutralizing antibody specificity in rabbits to one that was elicited during the course of natural infection. A better understanding of how vaccine-induced polyclonal neutralizing antibody responses compares to responses that developed in natural infection will improve our knowledge in designing better vaccine design strategies.The interplay between circulating virus variants and broadly cross neutralizing polyclonal antibodies developed in a subset of elite neutralizers is widely believed to provide strategies for rational immunogen design. In the present study, we studied the structural, antigenic and immunogenic properties of a novel soluble trimeric protein with near native pre-fusion conformation prepared using the primary sequence of an HIV-1 clade C We previously reported characterized genetic and neutralization properties of env sequences obtained from an Indian elite neutralizer (G37080) whose plasma antibodies demonstrated >90% neutralization breadth when tested against a large heterologous Env-pseudotyped virus panel overnight at 37\u00b0C for 16 to 24 h according to the manufacturer\u2019s protocol. Env proteins bound to magnetic beads were separated from unbound proteins using a DynaMag 15 magnet . Env protein bound beads were further incubated with blocking buffer at 37\u00b0C to block the unbound sites and the antigenic integrity of Env proteins were assessed by examining their ability to bind to different mAbs by flow cytometry . For depletion studies, plasma and serum samples were diluted to 1:50 in Dulbecco\u2019s modified Eagle\u2019s medium (DMEM) containing 10% fetal bovine serum (FBS), and 500\u03bcl of diluted plasma or serum were incubated with 20\u03bcl of Env protein coupled magnetic beads at room temperature for 45 min. Unbound plasma and serum antibodies were separated from bound antibody fraction using a DynaMag 15 magnet as described above. This step was repeated 4 to 5 times for gp120 and 1\u201312 times for SOSIP protein (1PGE-THIVC) towards facilitating efficient depletion. As a negative control, G37080 plasma antibodies were depleted with uncoated beads in parallel. In addition to ELISA, the percent depletion of G37080 plasma. The degree of depletion of the polyclonal serum and plasma antibodies were assessed by ELISA and TZM-bl neutralization assay as described previously [Purified soluble monomeric (92BR020 gp120) and trimeric 1PGE-THIVC (SOSIP gp140) proteins, in were used for the depletion of human plasma and rabbit serum neutralizing antibodies as described earlier . Brieflyeviously .Serum immunoglobulin G (IgG) was purified with a mixture of protein A/G affinity column. Purified IgG was digested for 6 hours at 37\u00b0C using 4% (w/w) liquid papain (Thermo Fischer Scientific) and digestion buffer . The digestion solution was collected, and Fab fragments were purified from undigested IgG and Fc-fragments using SEC . Final Fab yields were ~0.75\u20131.5 mg. Complexes were assembled with 10\u201315 \u03bcg of 1PGE-THIVC (SOSIP gp140) trimer and ~1 mg of purified polyclonal Fab, at room temperature for 18 hours. They were then purified using SEC with TBS as a running buffer and concentrated with 10 kDa cutoff Amicon ultrafiltration units. Samples were diluted in TBS to ~30 \u03bcg/ml and immediately deposited onto carbon-coated 400-mesh Cu grids (glow-discharged at 15 mA for 25 s), where they were then stained with 2% (w/v) uranyl formate for 30 s. For each sample, 116,958 to 250,000 individual particle images were collected and were subsequently submitted to 2D and 3D classification using Appion and ReliS1 Table(DOCX)Click here for additional data file.S1 FigPGT145 bnAb was taken as a negative control.(TIF)Click here for additional data file.S2 FigIn parentheses, highlighted residues and their respective colors are mentioned. The black arrows aid in providing the rotations done in model to present the image.(TIF)Click here for additional data file.S3 FigAmino acid numbering is made based on HXbc2 sequence. Amino acid residues in C3 and V4 that form key epitopes targeted by neutralizing antibodies induced in rabbits are highlighted. The glycan residue at the 362 position is underscored.(TIF)Click here for additional data file."} +{"text": "Veno-venous (VV) extracorporeal membrane oxygenation (ECMO) has become an integral part in the rescue therapy of severe acute respiratory distress syndrome (ARDS) and may be lifesaving in patients with refractory hypoxemia . VentilaPneumocystis jirovecii-associated pneumonia and severe ARDS we demonstrated earlier that a primarily awake ECMO strategy seems to be a promising strategy [In immunocompromised patients with ARDS who require ECMO support, the 6-month mortality exceeds 70% with a reported in-hospital mortality of 81% in patients following hematopoietic stem cell transplantation (HSCT) . In immuWe therefore hypothesized that awake ECMO support to avoid invasive ventilation in selected immunocompromised patients might yield improved outcomes. Here, we present a comprehensive summary of 18 immunocompromised patients who received awake ECMO support for management of ARDS at our institution between 09/2012 and 09/2020.2 despite maximal respiratory support by noninvasive ventilation (NIV). ECMO was initiated after a median of 1 (1\u20133) days following initial ICU admission and was carried out for 11 (9\u201318) days.The patient characteristics are shown in Table Eleven patients (61%) required secondary intubation after a median of 4 (2\u20136) days. The most common cause for secondary intubation was agitation stressing the critical role for delirium preventive strategies in these patients. The majority (4/6) of patients with agitation as primary cause for failing awake ECMO support developed agitation without any prior respiratory or circulatory deterioration. The choice of anxiolytic medication showed a trend toward a more frequent use of benzodiazepines in patients with agitation compared to all other patients (6/6 vs. 4/12), while low-dose morphine was used less frequently (2/6 vs. 7/12).28-day-, in hospital- and 6-month mortality rates were 44% (8/18), 50% (9/18) and 50%, respectively. In-hospital mortality was 29% (2/7) in solid organ transplantation patients and 50% (3/6) in hemopoietic stem cell transplantation patients. In-hospital mortality was 73% in patients who required secondary intubation and 14% in patients who did not require intubation while on ECMO support ).An exploratory analysis suggested several factor associated with later failure of an awake ECMO concept Table .Table 2PAlthough this study, to the best of our knowledge, represents the largest experience with awake ECMO in ARDS patients, conclusions are still limited by its small sample size and the uncontrolled nature. Despite these limitations, our findings support the notion that an awake ECMO strategy might be a viable treatment option for immunocompromised patients with severe ARDS, especially in those patients without overt multi-organ failure. Further studies are required to determine the possible role of the awake ECMO concept in patients with ARDS."} +{"text": "Arbona (2005) was incorrectly included as reference 15. As a result, all subsequent references are misnumbered. References 16\u201344 should be references 15\u201343.The third sentence of the Conclusions section should have cited reference 28 instead of 40 .The correct sentence should read: While the current results do not support the assumed academic benefits of physical activity, these results are consistent with another longitudinal investigation into the topic using COMPASS data [28].The fourth sentence of the Conclusions section should have cited reference 19 instead of 15 (the removed reference).The correct sentence should read: Furthermore, at least one additional small-scale study does appear to support the notion that brain health benefits of physical activity are realized in adolescents, despite unclear translation of such brain health benefits into overt academic achievement benefits [19]."} +{"text": "The University of New England College of Osteopathic Medicine (UNECOM) Geriatrics Education Mentors [GEM] program, established in 2014, pairs UNECOM students with older community living adults. GEM assignments focus on health review, medical humanities, and geriatrics training. Each year approximately 90 older adults participate in GEMs. In 2019, the GEM program was expanded with Geriatrics Workforce Enhancement Program (GWEP) grant funding to: include first year medical students, include 2 additional assignments (4 assignments over 10 months to 6 assignments over 18 months), and to create interprofessional student collaboration. In the new GEM Assignment 4: Medication Interactions/Contraindications, UNECOM students with their GEM compiled details on the GEM\u2019s medication list ; one of 4 Ms of Age Friendly Health Care. UNECOM students (84 pairs) were then assigned to UNE School of Pharmacy (SOP) students (42 SOP students had 2 UNECOM pairs) to conduct a \u201cLexicomp\u201d (App) medication interactions and Beers Criteria review. UNECOM students documented findings with the SOP student partner; discussed the processes of review with their GEM and the resultant findings; documented the GEM\u2019s questions and how the UNECOM student answered those questions; and discussed next steps for the GEM regarding options for different medications - especially follow up with their prescribing physician(s) for any noted interactions/contraindications. For GEMs with few medications, a mock medication list was assigned to ensure student experiences with medication reviews and GEM discussion. Although time intensive preparation is required, UNECOM & SOP students attained significant learning as did the GEM mentors."} +{"text": "With the approval of the injectables interferon-\u03b2 and glatiramer acetate a quarter of a century ago, the new era of multiple sclerosis (MS) treatment started . A decadNeurotherapeutics, a retrospective observational study representing a joint effort of 11 Italian MS centers compared effectiveness, tolerability, and safety of switching disease-modifying treatment in relapsing MS patients with a high JCV antibody index and at least 24 infusions from natalizumab to ocrelizumab, rituximab, or cladribine [As published in the April 2021 issue of adribine . The priadribine , 26.A recent observational study from Amsterdam analyzing 42 patients who stopped natalizumab and switched directly or indirectly to ocrelizumab obtained no evidence of disease activity (NEDA)-3 in 83% or 50%, respectively . Two patn\u2009=\u200913), continued disease activity (n\u2009=\u20096), presence of MRI disease activity (n\u2009=\u20094), and a switch to oral cladribine demonstrated effective disease suppression over a mean period of 9.7\u00a0months and no serious adverse events other than the expected lymphopenia [In a multicenter Swedish study of 256 relapsing MS patients who discontinued natalizumab because of JCV antibody positivity, rituximab was markedly superior to fingolimod in keeping clinical and MRI activity at bay over a period of 18\u00a0months . One sinphopenia .In aggregate, these observational studies provide evidence that high-efficacy drugs are effective and generally safe in a critical situation of MS management when treatment with natalizumab is discontinued. The results of an ongoing multicenter prospective open-label phase IV study examining the transition from natalizumab to ocrelizumab are expected by mid-2022 and will yield further data on which to base the important therapeutic decision.Finally, pilot studies reported abolition or marked diminution of PML risk in individuals receiving natalizumab at extended dosing intervals , 32. TheSupplementary file1 (PDF 508 KB)Supplementary file2 (PDF 508 KB)Supplementary file3 (PDF 508 KB)Supplementary file4 (PDF 508 KB)Below is the link to the electronic supplementary material."} +{"text": "Dear Editor,A 52-year-old Chinese male presented to the nephrology department in December 2017 with generalized swelling of the face and extremities and skin peeling. He had no previous history of hypertension, diabetes, or chronic kidney disease and was not taking any potentially nephrotoxic medications. He underwent gastrointestinal stromal tumor (GIST) resection in general surgery due to abdominal pain 2\u2009months before. Immunohistochemistry revealed that Vim (+), Pan-CK (\u2212), CD34 (+), S100 (\u2212), HHF35 (\u2212), SMA (\u2212), CD117 (+), Dog-1 (+), SDHB (+), P53 (\u2212), EMA (+), Ki67 index 10%. The histology results showed that gastrointestinal stromal tumors affected the entire thickness of the gastric wall, but there was no tumor metastasis in the perigastric lymph nodes (0/1) . To prevAs a tyrosine kinase inhibitor, imatinib is widely used in the treatment of chronic myeloid leukemia and GIST . StudiesPodocytes are terminally differentiated cells located outside the glomerular basement membrane, and the slit membrane formed by podocytes is the final barrier for glomerular filtration . After p"} +{"text": "Antibiotic prophylaxis (AP) in dentistry is recommended before dental procedures that involve manipulation of gingival tissue or periapical region, and perforation of oral mucosa in order to prevent a serious bacterial infection. The American Heart Association (AHA) Guideline recommends that the cardiac conditions with the highest risk of infective endocarditis (IE), for which AP is reasonable are: prosthetic cardiac valves, previous IE, congenital heart disease and cardiac transplantation with cardiac valvulopathy ,2,3. TheAn observational and descriptive study was conducted between June and August 2016 at Cl\u00ednica Dent\u00e1ria Egas Moniz (CDEM), a University Dental Clinic in the greater Lisbon Area. A total of 4000 patient records were analysed. 60 records were selected according to predefined inclusion criteria: patients submitted to endodontics and/or periodontology and/or surgery appointments, patients with IE/prosthetic cardiac valves/cardiac bypass/cardiac valvular disease/rheumatic fever or hip/knee joint prosthesis and patients who required AP before an invasive dental procedure. Clinical processes analysis were authorised by the patients through a declaration of informed consent. This study was authorised by the Clinical Director of CDEM and approved by Egas Moniz Ethics Committee.Periodontology patients had AP indication in: congenital cardiac disease (4/16), cardiac valvular disease (5/13), rheumatic fever (3/7), and prosthetic cardiac valves (7/8). Surgery patients had AP indication in: IE (2/2), congenital cardiac disease (4/12), cardiac valvular disease (4/6), rheumatic fever (2/4) and prosthetic cardiac valves (6/8). Endodontic records were not enough to have significant results to compare.There is some disparity between the AHA guideline and the attitudes of dentists towards AP indication . The use"} +{"text": "Opioids provide effective pain relief during flares of inflammatory bowel disease but are limited by serious side effects. We showed that acute administration of a novel pH-sensitive opioid agonist, NFEPP, had potent analgesic effects in inflamed acidified colonic tissues without opioid typical side effects. However, the effects of repeated application of NFEPP during the course of an acute flare of colonic inflammation are unknown.To assess the analgesic and side effect profile resulting from repeated NFEPP applications during the course of acute colitis.Acute colitis in C57BL/6 mice was induced via 2.5% dextran sulfate sodium dissolved in drinking water for 5 days. Mice were then randomly group assigned to either vehicle, fentanyl or NFEPP. Drugs and vehicle were administered sc. BID for 5 days (0.4 mg/kg), with a final injection on Day 6 (Day 1\u20136). Visceral nociception was evaluated by performing visceromotor responses (VMRs) to colorectal distension at different time points during colitis. Side effects were assessed using 1) a combination of oral contrast-enhanced abdominal CT-scans (before/after treatment) and defecation assessments to analyze gastrointestinal transit and isometric tension recordings to evaluate colon motility 2) pulse oximeter measurements to reveal cardiorespiratory effects. Inflammation was assessed by histological scoring, myeloperoxidase activity assay and tissue pH measurement of colon samples.NFEPP decreased VMRs to colorectal distension over the entire period of acute DSS colitis . However, strongest VMR inhibition was observed at Day 3 , concordant with the peak of inflammation . Treatment with NFEPP did not delay gastrointestinal transit nor fecal output whereas fentanyl decreased transit . Colonic contractile responses evoked by electrical field stimulation were reduced in fentanyl treated mice compared to NFEPP (10 Hz: p<0.05). Fentanyl significantly reduced oxygen saturation and caused a sustained reduction in heart rate at Day 2 whereas NFEPP did not affect oxygen saturation (p=0.95) and revealed minor, transient effects on heart rate which recovered after 60 min.Prolonged NFEPP administration effectively inhibits visceral nociception during acute colitis in a preclinical mouse model without altering gastrointestinal transit, colon motility and oxygen saturation.CCC"} +{"text": "To evidence that physical health monitoring during antipsychotic initiation and continued treatment within the Child and Family Clinic is current, as per the agreed Antipsychotic Medication Monitoring Schedule for Belfast Trust CAMHS (2015), supporting Quality Network for Community CAMHS(QNCC) accreditation.The Antipsychotic Medication Monitoring Schedule CAMHS(2015) was agreed by a working group of consultant psychiatrists and pharmacists, based on evidence from The Canadian Alliance for Monitoring Effectiveness and Safety of Antipsychotics in Children (CAMSEA), NICE Guidelines CG 185(2014), CG155(2013) and Maudsley Guidelines, and was to be located on the electronic system (PARIS).In January 2019, a list of all children/young people on antipsychotic medication was collated (n = 12). Presence of the monitoring schedule in the clinical notes or PARIS was recorded. The Electronic Care Record was reviewed for blood results and PARIS letters for documentation of physical health parameters and to identify documentation of risk/benefit review where monitoring was declined. Re-audit January 2020 (n = 9). Criteria:All patients commenced on antipsychotic medication will have baseline blood investigations and other physical health parameters documented as per the monitoring schedule. If monitoring was declined, the reason for this and indications for prescribing must be documented as a risk/benefit analysis.All patients on antipsychotic medication will be current with their physical health Monitoring Schedule.All patients will have their Monitoring Schedule completed in clinical notes or on PARIS.First cycle results (n = 12):Baseline bloods (or documented declined) = 92%, Baseline ECG (or documented declined) = 75%Complete monitoring bloods = 33%, Physical health monitoring parameters complete = 42%Monitoring schedule present in the notes and current = 42% (0% on PARIS).Initial Recommendations: Standardised recording of monitoring using PARIS clinic letters and the schedule in front of clinical notes; Baseline ECG mandatorySecond cycle results (n = 9):Baseline bloods (or declined) = 89%, Baseline ECG (or declined) = 67%Complete monitoring bloods = 44%, Physical health monitoring parameters complete = 56%Monitoring schedule present in notes and current = 38%, Present, not current = 50% (0% on PARIS).Lower numbers at re-audit limit interpretation.Further recommendations: Antipsychotic initiation checklist; Central bloods diary for clinicians; Antipsychotic care-pathway booklet, co-produced with young people, incorporating the monitoring schedule."} +{"text": "Rhizoctonia solani by Malik et al.(2019) should be Arshi Malik (2019)Bioinformation. 2019 15:227. PMID: 31285645The first author in \"Molecular docking and pharmacokinetic evaluation of natural compounds as targeted inhibitors against Crz1 protein in"} +{"text": "The aim of the study is to find out short term anatomical and visual outcomes of inverted internal limiting membrane flaps technique for large macular holes (base diameter>1000/vm), they were repaired with 25 gauge pars plana vitrectomy with brilliant blue assisted large inverted internal limiting membrane flap technique. Statistical analyses were performed using SPSS 19.0A descriptive cross-sectional study was conducted in a tertiary care hospital from January 2018 to December 2018 after ethical clearance from the institutional review committee. The study was done in 12 patients with idiopathic macular holes (base diameter>1000/vm. The mean age of patients was 68.75\u00b14.97 years. Postoperatively, mean best corrected visual acuity was 0.978 logMAR\u00b10.12. There were no postoperative complications. All the patients perceived decreased size of central scotoma.All twelve eyes had complete anatomical closure. Mean best corrected visual acuity preoperatively was 1.48 logMAR\u00b10.246. The mean macular hole base diameter was 1217.0\u00b1 196.77Inverted internal limiting membrane flaps for large macular holes is suitable method for closure of the very large hole, restoration of functional vision and decreased size of central scotoma. The inverted internal limiting membrane (ILM) flap is an effective technique for anatomical closure and improved visual function in large idiopathic full thickness macular hole (more than 400 /vm) patients.1Unilateral very large idiopathic macular holes (>1000 /vm) with inverted ILM flaps.2 In addition, par plana vitrectomy reduces vitreo-macular tangential tractions which Gass found vital causes for different stages of idiopathic macular holes.3Mahalingam et al. found 100 percent closure rate of large macular hole surgery in patients at a tertiary care hospital.The aim of the study is to find out short term anatomical and visual outcomes of inverted internal limiting membrane (ILM) flaps technique for large macular holes . Data was collected continuously during the study period. Study population were the patients who underwent surgery for large idiopathic macular holes. Patients with idiopathic large macular holes (MH base diameter>1000 /vm) were included in this study. Patient with MH with base diameter<1000 /vm, high myopic, traumatic, and media opacities were excluded from the study.Sample size was calculated using below mentioned formula and samples were collected using convenience sampling method.n = sample sizep = prevalence of macular hole closure with inverted internal limiting membrane flap technique, 95% (educated guess)q = 1-pd = margin of error, 13%Z = 1.96 at 95% CIwhere,After taking non-response rate of 10%, the total sample size was calculated to be 12.Patients underwent recording of best-corrected visual acuity (BCVA), fundus examination, SD OCT scan for measurement of macular hole base diameter at preoperative and follow up 1 month and 3 months. Each patients were asked whether they perceived decreased size of scotoma or not. Spectral Domain-Optical Coherence Tomography images were taken preoperative and postoperative 1 month and 3 months follow up to assess the anatomical outcome of surgery and best corrected visual acuity was recorded to know the functional outcome during each visitSelection bias and information bias was minimized as possible. Statistical analyses were performed using SPSS 19.0, point estimate at 95% CI was done along with proportion and frequency of the binary data.Twelve eyes had showed 12 (100%) macular hole closure rate with inverted ILM flap technique. Mean BVCA pre-operatively was 1.48 log MAR \u00b1SD 0.246. The mean macular hole base diameter was 1217.0 /vm (1036-1571 /vm). Post-operatively, mean BCVA was 0.978 log MAR \u00b1SD 0.12. The mean age of patients was 68.75\u00b14.97 years. Female were prepondent 8 (70%).1 Mahalingam P et al. showed similar results with inverted flap technique for repairment of minimal macular hole diameter greater than 800 /vm.2 As ILM is a basement membrane and part of muller cells, it induced glial cell proliferation to fill large macular holes with tissue over time. The thin rim attachment stabilized the peeled off ILM flap in the macular hole. The ILM flap induced proliferation of glial tissue to fill and secure the hole and to support the surrounding photoreceptor cells.4Advancement in micro instruments like vitrectomy cutter and intraocular ILM peeling forceps helped vitrectomy surgery with better outcome. Introduction of ILM staining dyes visualize the membrane more easily during ILM peeling. Among all Michalewska Z et al. firstly proved the usefulness of ILM flap to prevent the postoperative flat-open appearance of a macular hole repair and improvement of the functional outcomes of macular holes with a diameter greater than 400 /vm.5In our study, twelve eyes showed 100% macular hole closure rate with inverted ILM flap technique and other studies had similar closure rate for large macular hole.6 Guber J et al. showed upto 2 lines improvement in BCVA by first postoperative follow up.7Postoperative visual acuity was improved from 1.48 logMAR to 0.978 logMAR. Similar studies found improvement of mean visual acuity after large treated macular hole surgery with inverted ILM flaps.All the twelve patients perceived decreased the size of scotoma in the operated eye after surgery and were satisfied with postoperative functional visual improvement. Larger study group and longer follow-up period is required to further evaluate this method. Since this is a hospital based study, the findings of the study cannot be generalized to the population.There was improvement in anatomical, visual outcomes and alleviation of central scotoma for large macular hole (base diameter>1000 v m) surgery of inverted internal limiting membrane flaps technique for large macular holes (base diameter>1000/vm) surgery in patients compared to other studies.None."} +{"text": "Biosci Rep (2021) 41(3), https://doi.org/10.1042/BSR20204394) would like to correct The authors of the original article \u201cFive candidate biomarkers associated with the diagnosis and prognosis of cervical cancer\u201d ("} +{"text": "DISCUSSION/SIGNIFICANCE OF IMPACT: Targeting the Tie2 pathway is a new paradigm in regenerative medicine. Our designed constructs will enable us to generate high-affinity Tie2 agonists and antagonists as drugs to control angiogenesis, enabling tissue regeneration that recapitulates the biological architecture of the native tissue physiology, improving organ transplant outcome.OBJECTIVES/GOALS: Lack of blood vessels remains a major obstacle in tissue regeneration. Angiopoietin 1 and 2 modulate angiogenesis through the Tie2 receptor tyrosine kinase. Ang1 activates pAKT to promote endothelial cell survival while Ang2 antagonizes these effects. We aim to dissect the Ang/Tie2 pathway to uncover the molecular basis for these opposing effects. METHODS/STUDY POPULATION: Ang1 and Ang2 bind Tie2 via nearly identical F-domains (Fd). To investigate the molecular basis regulating the Tie2 pathway, we generated a series of computationally designed self-assembling protein scaffolds presenting F-domains in a wide range of valencies and geometries using Rosette Molecular Modeling Suite. We examined the protein kinase activation, cell migration, and blood vessel formation produced by the designed proteins in human umbilical vein endothelial cells. RESULTS/ANTICIPATED RESULTS: Two phenotypic classes were demonstrated by the number of presented F domains: scaffolds presenting 3 or 4 Fd have Ang2 like activity, upregulating pFAK and pERK but not pAKT and failing to induce cell migration and tube formation; scaffolds presenting more than 6 Fd have Ang1 like activity, upregulating the three signaling branches and enhancing cell migration and tube formation. Scaffolds with 8 or more Fd show superagonist activity, producing significantly stronger phenotypes than Ang1. These results suggest that Fd valency largely determines Ang1 vs Ang2 signaling outcomes, and our designed superagonists can outperform Ang1 in vascularization and wound healing. In"} +{"text": "Drosophila melanogaster pupal eye that has a highly stereotyped arrangement of cells. In addition, the pupal eye is postmitotic that allows for the study of tissue morphogenesis independent from any effects of proliferation. While the changes in cell morphology and organization that occur throughout pupal eye development are well documented, less is known about the corresponding transcriptional changes that choreograph these processes. To identify these transcriptional changes, we dissected wild-type Canton S pupal eyes and performed RNA-sequencing. Our analyses identified differential expression of many loci that are documented regulators of pupal eye morphogenesis and contribute to multiple biological processes including signaling, axon projection, adhesion, and cell survival. We also identified differential expression of genes not previously implicated in pupal eye morphogenesis such as components of the Toll pathway, several non-classical cadherins, and components of the muscle sarcomere, which could suggest these loci function as novel patterning factors.Tissue function is dependent on correct cellular organization and behavior. As a result, the identification and study of genes that contribute to tissue morphogenesis is of paramount importance to the fields of cell and developmental biology. Many of the genes required for tissue patterning and organization are highly conserved between phyla. This has led to the emergence of several model organisms and developmental systems that are used to study tissue morphogenesis. One such model is the Drosophila pupal eye is a postmitotic pseudostratified neuroepithelium that is organized into \u223c750 optical units known as ommatidia were used at 1:200. Dissected pupal eyes were imaged with a Leica DM5500 B fluorescence microscope and corresponding software.Pupal eyes were dissected and fixed as previously described . For 1\u00b0 Canton S pupae at the same time each day, and total RNA was extracted from three biological replicates using the ReliaPrep RNA Tissue Miniprep System as described . Quality control of sequence read outputs was performed using FASTQC lower than our predetermined threshold (q\u2009<\u20090.05). Gene ontology (GO) analyses were performed using the Gene Ontology Consortium (http://geneontology.org/). Volcano and scatter plots were created with R-statistical software (Between 50 and 70 eyes were dissected from 21 and 40 h APF escribed . Barcode Ensembl . Gene exsoftware (CRAN 20software .https://gbiomed.kuleuven.be/apps/lcb/i-cisTarget/index.php) (http://meme-suite.org/tools/ame) (https://flybase.org/) using batch download (https://flybase.org/download/sequence/batch/).Candidate regulatory transcription factors and transcription factor-binding motifs were identified with i-Cis Target analysis (ols/ame) using liRaw RNA-seq output files generated in this work are deposited under accession number GSE160441 in Gene Expression Omnibus.https://doi.org/10.25387/g3.12824789.Supplementary material is available at figshare DOI: Canton S eyes at 21 and 40 h APF indicating appropriate read depth for analysis (Supplementary Table S1). GO terms, identified by the Gene Ontology Consortium with the lowest P-values for all differentially expressed genes between 21 and 40 h APF included biological regulation (GO:0065007), cellular process (GO:0009987), regulation of biological process (GO:0050789), and regulation of cellular process (GO:0050794) (Supplementary Table S2). Of the 4636 genes, we identified the increased expression of 2383 genes at 40 h relative to 21 h APF and decreased expression of 2253 genes demonstrating no bias in the direction of regulation . The GO terms that were most significant for upregulated genes included nucleic acid metabolic process (GO:0090304), localization (GO:0051179), transport (GO:0006810), and establishment of localization (GO:0051234) (Supplementary Table S3). For downregulated genes, the most significant GO terms included regulation of biological process (GO:0050789), regulation of gene expression (GO:0010468), macromolecule metabolic process (GO:0060255), and regulation of metabolic process (GO:0019222) (Supplementary Table S4).To identify transcriptional changes during pupal eye morphogenesis, we dissected 40 h APF . These loci encompassed core components of several signaling pathways with established roles during Drosophila pupal eye development including the Notch, EGFR, Decapentaplegic (Dpp)/TGF\u03b2, and Planar Cell Polarity pathways and also identified signaling pathways that have not been interrogated for their role in fly eye development (Supplementary Table S5).Since development of the pupal eye is dependent on signal transduction, we assessed whether expression of core signaling proteins differed at 21 and 40 h APF. Our analyses identified differential expression of multiple loci broadly associated with signaling (GO:0023052) (N) and its ligand Delta (Dl) (Serrate (weary (wry) (O-fucosyltransferase 1 (O-fut1) (Notchless (Nle) (mind bomb 1 (mib1) (Suppressor of Hairless (Su(H)) (enhancers of split (E(spl)) genes that are transcriptional targets of Notch signaling (Supplementary Table S1).Notch signaling is vital during early pupal eye development for photoreceptor, cone and 1\u00b0 cell specification, patterning, and inducing cell death . At 40 hlta (Dl) (V\u00e4ssin Egfr (downstream of receptor kinase (drk), and SHC adaptor protein (Shc) (Son of sevenless (Sos) (Ras oncogene at 85D (Ras85D) (Raf oncogene (Raf) (Downstream of raf1 (Dsor1) (pointed (pnt) (Supplementary Table S5). While the expression of Egfr and many core RTK signaling components declined at 40 h APF, expression of several other RTKs and their ligands increased (Supplementary Table S5) suggestive of a continuous requirement for RTK-components in the eye. These included PDFG and VEGF receptor related (Pvr) (PDFG and VEGF receptor-related factor 1 (Pvf1) (Pvf2 (Pvf3 (branchless (bnl) (heartless (htl) were differentially expressed between 21 and 40 h APF including the secreted protease Spatzle-Processing Enzyme (SPE) (easter (ea) (Toll (Tl) , Toll-6 (encoded by Toll-6) and Toll-8 (encoded by tollo) form transheterophillic complexes that facilitate cell intercalation in the embryonic epithelium by stimulating actin reorganization (Supplementary Table S5) and studies that assess their role during pupal eye morphogenesis are an interesting avenue of future research.Our data also revealed differential expression of signaling proteins, which have not yet been implicated as regulators of pupal eye development. For example, we found that core components of the Toll signaling pathway (ter (ea) , the rec) . Toll siry gland . In addiabrupt (ab) at 40 h relative to 21 h APF, which negatively regulates the transcription of ecdysone target genes (Supplementary Table S1). We therefore reasoned that expression of documented Ecdysone transcriptional target genes would be reduced at 40 h APF and this was indeed the case for Edg91 (Eip71CD (Eip63E (ftz-f1 (ImpE2 and ImpL2 (Pep (Supplementary Table S6). However, the expression of other ecdysone response element genes increased, including Eip93F (Eip74EF (Eip78C (Eip75B (Supplementary Table S6). Since ecdysone signaling contributes to apoptosis in other Drosophila pupal tissues including the salivary gland, midgut, and muscle , it is possible that ecdysone response genes identified in our analyses of 21 h APF eyes contribute to or even initiate the apoptosis of lattice cells, which begins at around 18\u201320 h APF, correlating with increased ecdysone . These included hattifattener (haf), which increased in expression at 40 h relative to 21 h APF , and 10 e (Tutl) and incrSupplementary Table S7). For example, we identified increased transcription of Capulet (Capt), which inhibits actin filament growth (enabled (ena), which stimulates the addition of actin monomers and is required for axon elongation (cherrio (cher), a filament protein that crosslinks actin filaments (chickadee (chic), which encodes profilin (jitterbug (jbug), which encodes an actin cross-linking protein , a motor protein associated with axonal transport . For example, at 40 h APF, we detected decreased expression of the Ig domain adhesion molecules rst (hbs (sns (Kirre (2-fold change = 0.05). Since Notch signaling regulates the expression of hbs . We also identified decreased expression of shg (encodes E-cad) by 40 h APF (dachsous (ds) , suggestive of a prominent role for this cadherin during later stages of pupal eye development. Predictive bioinformatics analyses suggest that it binds to the members of the myosin family (head involution defective (hid) (reaper (rpr) (Death Regulator Nedd2-like caspase (Dronc) (Death related ICE-like caspase (Drice) (Supplementary Table S9). Our analyses also identified a significant decrease in the expression of Diap1 (SupplemSupplementary Table S10). These included rst, hbs, and sns, which are required for myoblast fusion during muscle development (Klarsicht (Klar), which is required for nuclei positioning in muscle cells (wings up A (wupA), which encodes the orthologue of Troponin I (Tropomyosin 2 (Tm2), which encodes a Tropomyosin that functions cooperatively with Troponin I during muscle contraction . To gain insight into common regulatory elements that might be utilized to facilitate changes in gene expression, we used full i-Cis target analysis (Supplementary Tables S5\u2013S10). These analyses identified transcription factor-binding sites that were common to genes associated with signaling, axon guidance, biological adhesion, or muscle structure development, suggesting that these processes are regulated by a limited number of transcription factors (Supplementary Table S12). In contrast, we did not identify recurrent transcription factor regulatory motifs in the groups of genes associated with ecdysone signaling or cell survival that changed in expression from 21 to 40 h APF. This suggests that in the eye the genes associated with cell survival/apoptosis and ecdysone signaling are not a part of regulatory networks but instead their expression is regulated by multiple transcription factors.Our transcriptome analyses identified 4636 differentially expressed loci that contribute to multiple biological processes (Trithorax-like (Trl) (Supplementary Table S12 and Supplementary Table S13) . This change in Trl expression likely contributes to the very different transcriptional profile of the eye at 40 h APF (Supplementary Table S1).The conserved GAGA transcription factor, ble S13) . In a prSupplementary Table S12 and zld led to small and deformed eyes as a putative regulator of genes associated with axon guidance and muscle structure as well as Zelda (Zld), which may contribute to the expression of genes associated with axon guidance and adhesion (Supplementary Table S5), ecdysone response targets (Supplementary Table S6), regulators of axon guidance (Supplementary Table S7), adhesion (Supplementary Table S8), and cell survival (Supplementary Table S9). In addition, we identified numerous novel gene expression changes that have not been studied in the context of pupal eye morphogenesis. These included components of the Toll pathway (Supplementary Table S5), non-classical cadherins (Supplementary Table S8), and numerous proteins required for muscle development and structure (Supplementary Table S10). We anticipate that these data will be a rich resource for future research on pupal eye morphogenesis and, given the highly conserved nature of many genes associated with tissue patterning, for the broader morphogenesis field as well.To conclude, the pupal eye is an effective model system to use to interrogate processes required for the patterning or organization of epithelia as well as pathways that lead to cell differentiation, and photoreceptor morphogenesis and axon projection. However, while many of the morphological changes associated with these events have been well documented, less is known of the corresponding transcriptional changes that drive them. Here, we compare the transcriptomes of pupal eyes at two distinct stages of development. We identified changes in the expression of loci that are documented regulators of pupal eye development such as components of Notch and EGFR signaling pathways ("} +{"text": "Optimal blood pressure (BP) control in nursing home residents is controversial and this population has been excluded from trials. We evaluated the associations of BP level with cardiovascular (CV) events and all-cause mortality across antihypertensive medication categories in Veterans Affairs (VA) nursing home residents. Data for 18,589 residents aged 65 years and older was obtained from the VA Corporate Data Warehouse from October 2006 through September 2017. Baseline systolic BP (SBP) and diastolic BP (DBP) were divided into categories and analyses were stratified by antihypertensive therapy . Over a median follow-up of 1.8 years, CV events occurred in 3,519 (19%) residents and 15,897 (86%) residents died. In participants on no BP medications, high SBP (>150 mmHg) was associated with a greater risk of CV events compared with normal SBP (110-130mmHg). By contrast, in participants on \u22652 BP medications, the subgroup with low SBP (<110 mmHg) had a higher CV risk . For DBP, in participants without BP medications, there were no differences in CV risk across DBP subgroups. Whereas among those on 1 or \u22652 medications, DBP <60 mmHg was associated with a higher CV risk compared with normal DBP (70-80 mmHg). Participants with low SBP (<110 mmHg) and DBP (<70 mmHg) had an increased mortality risk regardless of the number of medications. These findings suggest a potential risk of low BP among nursing home residents on multiple antihypertensive medications."} +{"text": "Medical residents need training to assess social determinants of health (SDOH) related to chronic conditions. We created a checklist to identify SDOH affecting residency clinic patients\u2019 ability to manage chronic conditions. The tool: 1) involves resident training; 2) provides decision support checklist; 3) influences patient activation; and 4) increases provider and patient communication through shared decision making. Action Planning Guide checklist (APG) includes questions pertaining to SDOH preventing patients from managing their chronic conditions and actions patients will take. Areas identified are discussed between patient and resident, increasing patient activation. The clinic\u2019s nurse care facilitator guides referrals to community-based resources. Fifty-two patients were enrolled, with 75% of patients responding they would like to be better managers of their chronic conditions. This information is used to develop patient\u2019s goals of care. Over 90% of patients said their conditions affect their lives and discussed ways better to care for themselves. Over 80% discussed medication management, health goals to improve their quality of life, and made a plan that maps out ways to reach their goals. All of these are essential for achieving positive health outcomes for older patients with chronic conditions. These attributes promote effective patient/provider partnerships. Seventy referrals were made; food through 2-1-1 (47%); monthly commodity food program (30%); utility payments (11%), and transportation (9%). Twenty-seven referrals were made to agencies serving older adults; 25 to the local AAA information and assistance services, and 2 to Senior Project Fresh Voucher Program."} +{"text": "In the article titled \u201cLong-Term Effectiveness of Oral Ferric Maltol vs Intravenous Ferric Carboxymaltose for the Treatment of Iron-Deficiency Anemia in Patients With Inflammatory Bowel Disease: A Randomized Controlled Noninferiority Trial\u201d, the legends for Figures 3 and 4 were incorrect.The incorrect legends read as follows:P values shown for test of null hypothesis of inferiority in risk difference with noninferiority margin of 20%.\u201d.\u201cFigure 3. Mean change in Hb over 52 weeks of treatment with oral ferric maltol or IV ferric carboxymaltose . P values shown for test of null hypothesis of inferiority in risk difference with noninferiority margin of 20%.\u201d.\u201cFigure 4. Patients achieving \u22651 and \u22652 g/dL increases, and normalization of Hb concentration between baseline and week 12 (ITT population with multiple imputation). The legends should read as follows:P values shown for least-squares mean change from baseline, difference between groups.\u201d.\u201cFigure 3. Mean change in Hb over 52 weeks of treatment with oral ferric maltol or IV ferric carboxymaltose (ITT population). P values shown for risk difference between groups.\u201d.\u201cFigure 4. Patients achieving \u22651 and \u22652 g/dL increases, and normalization of Hb concentration between baseline and week 12 (ITT population with multiple imputation). These errors have been corrected online and in print."} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-021-91087-5, published online 04 June 2021Correction to: The original version of this Article contained errors in the Figure legends of Figure 3 and Figure 4. The legends of these Figures were inadvertently switched.The legend of Figure 3:\u201cmRNA expression of genes in bronchial epithelium using GSE67472 dataset to compare healthy controls (n\u2009=\u200943) to Th2-high asthmatics (n\u2009=\u200940) and Th2 low asthmatics (n\u2009=\u200922).\u201dnow reads:\u201cmRNA expression of the ten genes in bronchial epithelium using, GSE76227 transcriptomic dataset that contains the expression data of 190 bronchial biopsies (BB) and epithelial brushing (BRUSH) from Unbiased BIOmarkers in Prediction of REspiratory Disease outcomes (U-BIOPRED) Project. The normalized gene expression of each of the identified genes was extracted and compared between different subgroups. The datasets were subdivided into nonsevere asthmatic (MAS), severe asthmatics (SAS) vs. oral steroid users (OS).\u201dThe legend of Figure 4:\u201cmRNA expression of the ten genes in bronchial epithelium using, GSE76227 transcriptomic dataset that contains the expression data of 190 bronchial biopsies (BB) and epithelial brushing (BRUSH) from Unbiased BIOmarkers in Prediction of REspiratory Disease outcomes (U-BIOPRED) Project. The normalized gene expression of each of the identified genes was extracted and compared between different subgroups. The datasets were subdivided into nonsevere asthmatic (MAS), severe asthmatics (SAS) vs. oral steroid users (OS).\u201dnow reads:\u201cmRNA expression of genes in bronchial epithelium using GSE67472 dataset to compare healthy controls (n\u2009=\u200943) to Th2-high asthmatics (n\u2009=\u200940) and Th2 low asthmatics (n\u2009=\u200922).\u201dThe original Article has been corrected."} +{"text": "Aortic diameter measurements in patients with a thoracic aortic aneurysm(TAA) show wide variation. There is no technique to quantify aorticgrowth in a three-dimensional (3D) manner.To validate a CT-based technique for quantification of 3D growth based ondeformable registration in patients with TAA.Patients with ascending and descending TAA with two or more CTangiography studies between 2006 and 2020 were retrospectivelyidentified. The 3D aortic growth was quantified using vasculardeformation mapping (VDM), a technique that uses deformable registrationto warp a mesh constructed from baseline aortic anatomy. Growthassessments between VDM and clinical CT diameter measurements werecompared. Aortic growth was quantified as the ratio of change in surfacearea at each mesh element (area ratio). Manual segmentations wereperformed by independent raters to assess interrater reproducibility.Registration error was assessed using manually placed landmarks.Agreement between VDM and clinical diameter measurements was assessedusing Pearson correlation and Cohen \u03ba coefficients.n = 26), descendingaorta (n = 10), or both (n = 2). VDMwas technically successful in 35 of 38 (92%) patients and 58 of 68intervals (85%). Median registration error was 0.77 mm . Interrater agreement was high for aorticsegmentation (Dice similarity coefficient = 0.97 \u00b1 0.02) andVDM-derived area ratio . There was strong agreement between peak area ratio values anddiameter change. VDM detected growth in 14 of 58 (24%) intervals. VDMrevealed growth outside the maximally dilated segment in six of 14 (36%)growth intervals, none of which were detected with diametermeasurements.A total of 38 patients were evaluated , with TAAinvolving the ascending aorta (Vascular deformation mapping provided reliable and comprehensivequantitative assessment of three-dimensional aortic growth and growthpatterns in patients with thoracic aortic aneurysms undergoing CTsurveillance.Published under a CC BY 4.0 licenseOnline supplemental material is available for thisarticle.Wieben in this issue.See also the editorial by Vascular deformation mapping, a deformable image registration-based technique,enabled reliable comprehensive assessment of the degree and extent ofthree-dimensional growth among patients with a thoracic aortic aneurysmundergoing CT surveillance.\u25a0 In a retrospective analysis of 38 patients with thoracic aorticaneurysm on CT scans, vascular deformation mapping (VDM) was technicallysuccessful in 35 of 38 (92%) patients and 58 of 68 intervals (85%).\u25a0 VDM was used to detect growth in 14 of 58 (24%) intervals, withsix detected outside of the maximally dilated segment, none of whichwere detected with clinical diameter measurements based on results of CTangiography.r = 0.85,P < .001).\u25a0 VDM-derived measurements of aortic surface area change had lowinterrater variability (bias = 0.0); peak area ratio values and diameterchange showed strong agreement (Thoracic aortic aneurysm (TAA) is common and is increasing in prevalence worldwide,with approximately 3% of patients older than 50 years having a dilated thoracicaorta \u20133 and reTo overcome these limitations, prior research has described the feasibility of amedical image analysis technique, termed vascular deformation mapping (VDM), in 3Dassessment of aortic growth using deformable image registration techniques ,13.This(a) to determineperformance of the VDM algorithms in a cohort of patients with TAA undergoingimaging surveillance that included assessment of reproducibility and identificationof sources of error in the analysis workflow and (b) tocharacterize unique patterns of 3D aortic growth observed in patients with TAA andto assess the agreement of VDM analysis with standard diameter measurements.This study focused on two primary objectives: Appendix E1 (online). Clinical anddemographic information was collected through chart review. Maximal diametermeasurements of the thoracic aorta were recorded from clinical CT reports. Ofnote, aortic measurements at our center are performed in a 3D laboratory bytrained technologists using standardized measurement protocols and a centerlinemeasurement technique and were compliant with the Health Insurance Portability andAccountability Act. We used electronic medical records search software developedat our institution to identechnique .(a)segmentation of the thoracic aorta on CT angiography images from scans acquiredat two different time points, with the first time point considered the fixedimage and the second time point considered the moving image;(b) image preprocessing steps including cropping andclamping voxels with negative attenuation values (in Hounsfield units) at 0 toavoid the adjacent lung influencing the registration and dilation of aorticmasks by three voxels to ensure inclusion of the wall; (c)rigid registration to approximately align the two CT angiographic images ((d) implicit alignment of the aortic centerline using ahighly regularized multiple-image multimetric deformable registration thatapplies a penalty term to enforce rigid movement of voxels within the aorticsegmentation but allows deformation of the periaortic voxels ((e) multiresolutionmultimetric B-spline deformable image registration using mutual information with10-mm grid spacing and a bending energy penalty of 100 ((f) generation of a polygonal mesh ofthe aortic surface at baseline (fixed) geometry; (g)translation of baseline aortic mesh vertices using the deformation fieldcalculated in step 5; and (h) quantification of deformation asthe ratio of surface area change at each triangular mesh element (termed arearatio) with color visualization in Paraview 5.9.0 (Kitware). VDM analysis takesapproximately 20 minutes on a standard high-performance PC with parallelization.A simplified schematic overview of the VDM analysis pipeline is presented inThe VDM analysis pipeline for measurement of 3D aortic growth uses deformableimage registration to quantify deformation of the aortic wall between two CTangiograms. The VDM analysis includes several steps: versity) ;(d) impc voxels ; (e) muly of 100 ; (f) genManual aortic segmentation was used in the VDM workflow to create aortic masksand is thus a potential source of variability. While all CT angiograms weresegmented by a rater with 4 years of experience with aortic image analysis(I.B.H.), we had an additional rater with 5 years of experience (T.M.J.v.B.)perform segmentations on 45 randomly selected CT angiography intervals toinvestigate the influence of manual segmentation variability on VDM output.Raters segmented the thoracic aorta from the root to just beyond the celiacaxis, including the proximal arch vessels, using segmentation software .Appendix E1 (online).We adopted a multistep quality assurance protocol to evaluate the validity ofeach VDM output, with quality assurance steps performed by a researcher with 15years of experience with cardiovascular CT (N.S.B.). The quality assuranceprotocol involved visual confirmation of segmentation and registration accuracyusing dual-color plots to ensure overlap of the aortic wall after the finaldeformable registration step; specific steps in the quality assurance protocolare described in To assess registration accuracy, landmarks were manually placed along the aorticwall by a senior researcher with 15 years of cardiovascular CT experience(N.S.B.). Landmark registration error was determined by calculating theEuclidean distance between homologous points after deformable transformation.Conserved anatomic landmarks, such as branch points and intimal calcifications,were used to place aortic landmarks across serial CT angiograms. Deformableregistration was performed using VDM parameters in both the forward and thereverse direction and using all possible combinations of CT intervals for eachpatient.P< .05 was indicative of a significant difference for all statisticaltests. Statistical analyses were performed using Stata 14.0 (StataCorp).Continuous variables are reported as mean \u00b1 standard deviation fornormally distributed data, as median and interquartile range (IQR) for nonnormalcontinuous variables, and as frequencies for categorical variables. Normalitywas assessed using the Shapiro-Wilk test. Pearson correlation coefficient wasused to assess correlation between continuous variables. Binary categories werecreated based on published data on reducibility of clinical diametermeasurements \u201310, withn = 3), excessive motion orstair-step artifacts (n = 2), streak artifacts from densesuperior vena cava contrast (n = 2), and streak artifactsrelated to superior vena cava cardiac implantable electronic device leads(n = 3). Examples of error cases are shown inFigure E1 (online).Of the 50 patients undergoing imaging surveillance of TAA on CT scans, five wereexcluded for lack of electrocardiographically gated CT acquisition, one wasexcluded for lack of thin-section reconstructions, two were excluded for pooraortic opacification, two were excluded for interval surgical aortic repair, andthree were excluded for severe motion artifact. A total of 38 unique patientsencompassing 105 CT angiograms and 68 surveillance intervals were selected foranalysis. Among the 38 patients included for analysis, 3D growth mapping withVDM was successful in 35 (92%), and VDM analysis was successful in 58 of 68(85%) surveillance intervals. Reasons for registration failure identifiedincluded irregular section intervals in source Digital Imaging andCommunications in Medicine images . The majority of TAAsinvolved the ascending aorta and were considereddegenerative in origin . Approximately one-thirdof patients (11 of 38) had a history of prior aortic repair and were undergoingpostsurgical surveillance. Complete patient characteristics are shown in theThe mean patient age was 69 years \u00b1 9 , andmost patients were female .A total of 199 unique landmarks were manually placed at discrete anatomiclocations along the aortic wall in 79 CT angiograms with a mean of 7.2 landmarksper patient. Considering all registration combinations, a total of 1021point-pairs were used to assess landmark registration error. The medianregistration error was 0.77 mm and anaverage Hausdorff distance of 0.12 mm \u00b1 0.20 . When comparing the interrater agreement of area ratio values betweenapproximately 5.4 million homologous surface elements, we found no bias (bias =0.0), narrow limits of agreement , and growth was detected with VDM in 14 of 58 (24%)intervals (defined as peak area ratio \u22651.2). VDM analysis clearlydepicted aortic growth in common TAA locations including the ascending aorta, descendr = 0.85; 95% CI: 0.75, 0.91;P < .001) between peak area ratio values and thechange in maximal aortic diameter with clinical CT thoracic aortic aneurysm growth quantification using vasculardeformation mapping (VDM) in a clinical cohort of patients with variousmanifestations of thoracic aortic aneurysm commonly encountered in clinical practice. In summary, we found thatVDM analysis was technically successful in 85% of the evaluated intervals and thatthe most common reasons for failure of the VDM analysis included artifacts relatedto streak and motion artifacts at the ascending aorta. Despite small degrees ofinterrater variability in aortic segmentations, the final surface area ratio fromVDM analysis showed excellent interrater agreement. In addition to quantifying 3Daortic growth in the maximally dilated segment, VDM identified additional regions ofgrowth outside the primary aneurysmal segment in approximately one-third ofpatients. Lastly, while VDM demonstrated agreement with diameter growth assessmentsin the majority of cases (89%), the 3D nature of VDM allows for a more comprehensivedepiction of the extent and distribution of growth along the aortic surface than ispossible with diameter measurements.Aortic diameter is the current metric used to assess growth and determine candidacyfor surgical repair. However, diameter measurements vary and are limited in theirability to enable prediction of progressive growth and acute complications, such asaortic dissection ,11. AsseBeyond providing a reproducible assessment of growth, the 3D nature of VDM allows fora more comprehensive evaluation than two-dimensional aortic diameter measurements.Quantitative mapping of TAA growth allows for investigation of unique parameters that are otherwise unable tobe easily captured. While VDM represents one of the first techniques forquantitative mapping of disease progression in TAA, similar image analysistechniques using deformable image registration have been used to phenotype andassess progression of diseases of the lungs ,20, braiOur study had several limitations. First, we did not systematically investigate theassociation of VDM metrics with patient outcomes, which will require larger cohortswith longitudinal follow-up. Second, while VDM analysis was technically successfulin 92% of surveillance intervals, the technique is susceptible to errors in thepresence of streak and motion artifacts. Thus, the performance of VDM may besuboptimal at centers that do not routinely use electrocardiographic gating andthose that have older-generation CT scanners with narrower detector arrays, limitinggeneralizability. Third, VDM analysis currently requires more time than diametermeasurement ; however, the overall analysis time can be mitigated by deep learningtechniques for automated aortic segmentation . Lastly,In conclusion, vascular deformation mapping (VDM) is a reproducible method forcomprehensive three-dimensional (3D) quantification of longitudinal aortic growth ina heterogeneous cohort of patients with thoracic aortic aneurysm. VDM analysisyielded reliable growth assessments in most surveillance intervals with excellentinterrater reproducibility. Failure of this new method was predominantly related tostreak and motion artifacts. Accurate quantitative 3D assessments of aortic growthmay enable a more nuanced assessment of patient risk, disease phenotypes, and growthtrajectories and may serve to better inform surveillance intervals, treatmentdecisions, and outcomes in patients with thoracic aortic aneurysm (TAA). However,given the low complication rate and slow growth of TAA, defining the prognostic andclinical importance of VDM measurement changes in aortic surface area requiresfurther investigation in larger cohorts of patients with long-term follow-up."} +{"text": "Clostridium tyrobutyricum is recognized as the main causative agent of late blowing defect\u2014severe spoilage of hard and semihard cheeses. In this work, we present the draft genome sequences of 12 C. tyrobutyricum strains isolated from raw milk and cheese. Clostridium tyrobutyricum is an endospore-forming anaerobic bacterium recognized as the main causative agent of late blowing defect (LBD)\u2014severe spoilage of hard and semihard cheeses, leading to great financial losses for the dairy industry isolated from raw milk samples , and single colonies were grown anaerobically at 37\u00b0C overnight in reinforced clostridial medium (RCM) (Frozen stocks (\u221280\u00b0C in 20% glycerol) of um (RCM) . Genomicum (RCM) was usedum (RCM) was usedum (RCM) was usedum (RCM) , discardum (RCM) . All reaum (RCM) and wereum (RCM) to deterum (RCM) . Unless PRJNA673079.The draft genome sequences and raw reads are available in GenBank under the BioProject accession number"} +{"text": "Informative serum biomarkers for monitoring inflammatory activity and treatment responses in axial spondyloarthritis (axSpA) are lacking. We assessed whether Lipocalin 2 (LCN2) and Oncostatin M (OSM), both having roles in inflammation and bone remodeling, may accurately reflect chronic joint inflammation and treatment response in axSpA. Previous reports in animal models showed involvement of LCN2 and OSM in joint/gut inflammation. We asked whether they also play a role in human axSpA.We analyzed a longitudinal observational axSpA cohort (286 patients) with yearly clinical assessments and concurrent measurements of serum LCN2 and OSM (1204 serum samples) for a mean of 4 years. Biomarker levels were correlated with MRI scoring and treatment response.p = 0.0005 and 0.005 for LCN2 and OSM respectively), suggesting that LCN2/OSM outperforms CRP as reflective of SIJ inflammation. We observed both concordant and discordant patterns of LCN2 and OSM in relationship to back pain, the cardinal clinical symptom in axSpA. Twenty-six percent (73/286) of the patients remained both clinically and serologically active (CASA). Sixty percent (173/286) of the patients became clinically quiescent, with back pain resolved, but 53% (92/173) of them were serologically active (CQSA), indicating that pain control may not indicate control of joint inflammation, as reflected by positive MRI imaging of SIJ. With respect to treatment responses, transient elevation of LCN2 or OSM over time was predictive of better response to all treatments.Persistent and transient elevation of LCN2 and OSM were observed in axSpA patients. Persistent elevation of LCN2 or OSM, but not CRP, correlated with sacroiliac joint (SIJ) MRI SPARCC scores (Pearson\u2019s correlation In axSpA, persistent LCN2 and/or OSM elevation reflects chronic SIJ inflammation and suboptimal treatment response. In our cohort, half of the currently deemed clinically quiescent patients with back pain resolved continued to demonstrate chronic joint inflammation. LCN2 and OSM profiling outperforms CRP as a predictive measure and provides an objective assessment of chronic local inflammation in axSpA patients.The online version contains supplementary material available at 10.1186/s13075-021-02521-y. Most of the patients with discordant response are CQSA . Only two patients with discordant response are CASQ . For patients with persistent LCN2 elevation (L++), profiling indicated that both concordant and discordant patterns were observed. Patients with concordant response were predominantly CQSQ] as defined by normal LCN2, undetectable OSM, and low back pain scores. The remaining 42% (26/62) showed discordant treatment response, having the CASQ pattern as both LCN2 and OSM were persistently normal, but back pain persisted are deemed responders and 22% [14/65] were C++ and C+ respectively. C++ patients had higher LCN2 levels . C++ patients have higher and lower percentage of CASA and CQSQ patients respectively in L++ patients , a larger study is warranted. Though MRI is currently the most sensitive tool for the detection of joint inflammation, it is not without limitations such as false positives for patients with low back pain. The use of LCN2 and OSM monitoring serves as a pre-screen to determine whether the costly MRI is needed to confirm findings from LCN2/OSM profiling.Secondly, the first demonstration that LCN2 and OSM levels correlated with MRI SPARCC SIJ scores and thus reflect SIJ inflammation, the cardinal feature of axSpA. This report is focused on the relationship of LCN2 and OSM and chronic inflammation. This is the reason why we chose patients with no spinal ankylosis (mSASS=0) to analyze the correlation of LCN2 or OSM levels with MRI inflammation scores. As low numbers of patients with single pathway involvement and MRI assessment were available for this study had back pain resolved but LCN2 or OSM remained elevated. In our MRI inflammatory scores vs serological level correlations, 88% (7/8) CQSA patients had positive MRI SIJ SPARCC scores, and the single CQSA patient who had negative MRI SIJ SPARCC scores had positive Berlin spine scores, suggesting that persistent LCN2 or OSM levels are associated predominantly with SIJ inflammation were SA and 9 of them (11%) had mSASS >50. This is in contrast to only 3% (3/92) CQSQ patients who had mSASS>50 , indicating that though back pain may be controlled (CQ), the disease may progress if LCN2/OSM continues to be elevated (SA). Thus, LCN2 and OSM profiling provides personalized and more effective treatment.Fourthly, current axSpA management focuses on symptom (back pain) control. Of the 173 CQSQ, although about half of them are CQSA with back pain resolved but OSM elevation persisted and with normal LCN2 (Ln). Pearson's correlation coefficient test was used. Table S1. Demographics of axSpA patients in different categories. Table S2. Summary of different categories of patients with the involvement of different pathway(s). Table S3. Sensitivity, specificity, positive and negative predictive values using different LCN2 cutoffs. Table S4. Treatment outcome in patients with LCN2 pathway alone (TNFi vs no TNFi). A. Comparison of OSM negative patients treated with TNFi: patients with persistent LCN2 elevation (L++) vs. transient LCN2 elevation (L+) vs. normal LCN2 (Ln). B. Comparison of OSM negative patients never received TNFi treatment: L++ vs. L+ vs. Ln patients. One-way analysis of variance followed by Bonferroni\u2019s multiple comparison test and Pearson\u2019s chi square test were used. Table S5. Treatment outcome in patients with OSM pathway alone (TNFi vs no TNFi). A. Comparison of patients with normal LCN2 treated with TNFi: patients with persistent OSM elevation (O++) vs transient OSM elevation (O+). LCN2 and OSM levels were compared between these two patient groups. B. Comparison of patients with normal LCN2 and never received TNFi treatment: O++ vs O+ patients. LCN2 and OSM levels were compared between these two patient groups. Table S6. The use of 95%CI of LCN2 and OSM to predict signature of axSpA patients. Table S7. Prevalence of patients having mSASS>50 in CQSA vs CQSQ subgroups."} +{"text": "Hereditary causes of ovarian cancer include Lynch syndrome, which is due to inherited pathogenic variants affecting one of the four mismatch repair genes involved in DNA repair. The aim of this study was to evaluate tumour mismatch repair deficiency and prevalence of Lynch syndrome in high-risk women referred to the Manchester Centre for Genomic Medicine with ovarian cancer over the past 20 years.MLH1 promoter methylation testing followed by constitutional testing for Lynch syndrome.Women with ovarian cancer diagnosed before the age of 35 years and/or with a suggestive personal or family history of Lynch syndrome cancers underwent tumour testing with immunohistochemistry for mismatch repair deficiency and, where indicated, MLH1 promoter hypermethylation, and 18 of the remaining 24 underwent constitutional testing for Lynch syndrome. A further 15 women with mismatch repair proficient tumours underwent constitutional testing because of a strong family history of Lynch syndrome cancers. Pathogenic variants were identified in 9/33 (27%) women who underwent constitutional testing, aged 33\u201359 years (median 48 years), including one whose tumour was mismatch repair proficient. Most Lynch syndrome tumours were of endometrioid histological subtype.In total, 261 ovarian cancers were tested and 27 showed mismatch repair deficiency by immunohistochemistry. Three of 7 with MLH1 loss showed Tumour mismatch repair deficiency identified by immunohistochemistry is a useful prescreen for constitutional testing in women with ovarian cancer with personal or family histories suggestive of Lynch syndrome. BRCA1 and BRCA2 with as many as 22% of women with high grade serous ovarian cancers (HGSOC) carrying pathogenic variants in these genes.MSH2, MLH1, MSH6 and PMS2.Ovarian cancer is the seventh most common malignancy worldwide and the most lethal gynaecological cancer.Since the discovery of the MMR genes in 1993\u20131994, clinicians have tried to target constitutional testing for LS to those at highest risk. The Amsterdam criteria were developed in 1991,We have evaluated our prescreening strategy with IHC in women referred to the regional genetics department with possible LS-associated ovarian cancer from 2000 to 2020 and assessed the identification of constitutional MMR pathogenic variants.Women referred to the regional genetics department in Manchester with ovarian cancer and concerns about the possibility of LS provided consent for tumour and if indicated constitutional testing. Most women had a history of another LS-related cancer in themselves or another family member . However, some were selected based on diagnosis at <35 years of age.IHC for the four MMR proteins was performed in the clinical pathology laboratory using the automated Ventana BenchMark ULTRA IHC\u2044ISH staining module and the OptiView, 3\u2019diaminobenzidine V.5 detection system according to standard clinical protocols. Tumour epithelial MMR expression was scored by two expert independent observers using stroma as internal control and as described previously.MLH1 promoter methylation testing was performed on tumours showing loss of MLH1 on IHC. Extracted DNA was bisulfite converted and then amplified with bisulfite specific primers in triplicate. A region of the MLH1 promoter containing four CpG dinucleotides whose methylation status is strongly correlated with MLH1 expression were sequenced using a pyrosequencer (PSQ 96MA). Two independent scientists interpreted the pyrograms. \u2018Hypermethylation\u2019 was described as\u00a0>10%\u2009mean methylation across the four CpG dinucleotides on a minimum of two of three replicate analyses. In addition to promoter methylation analysis, testing was carried out for the BRAF c.1799T>A variant in some cases.Reflex Extracted DNA underwent sodium bisulfite conversion using the Epitect Plus FFPE kit . The MSI analysis system V.1.2 used fluorescent-labelled primers to coamplify seven markers, including five mononucleotide-repeat markers , and two control penta-nucleotide-repeat markers (Penta-C/Penta-D). MSI status was reported as microsatellite stable (MSS) where all five mononucleotide loci between tumour and matched normal tissue were identical; MSI-low (MSI-L) where there was discordance in one mononucleotide locus and MSI-high (MSI-H) where two or more mononucleotide loci were discordant.MLH1, MSH2 and MSH6 were amplified using long range PCR followed by next generation sequencing using Illumina SBS v2 2\u00d7150\u2009bp chemistry on an Illumina MiSeq. The whole coding region, intronic flanking sequences to\u00b115\u2009bp and known splicing variants of MLH1, MSH2 and MSH6 were analysed (minimum 100\u2009x coverage depth). Variant identification and calling was via an in-house bioinformatic pipeline. Reported sequence changes and regions with <100\u00d7\u2009coverage were retested via Sanger sequencing using BigDye V.3.1 chemistry. Copy number analysis to detect large genomic rearrangements affecting the three MMR genes was performed using MLPA MRC-Holland probe mixes: P003-D1 MLH1/MSH2 and P072-C1 MSH6. Variant nomenclature followed Human Genome Variation Society guidelines (http://www.hgvs.org/vamomen) using reference sequences: LRG_216, t1(MLH1); LRG_218, t1(MSH2); LRG_219, t1(MSH6). Exons were numbered consecutively starting from exon 1 as the first translated exon for each probe mix. Cases with PMS2 protein loss, normal MLH1 methylation and no path_MLH1/MSH2/MSH6 variant underwent path_PMS2 analysis at the regional specialist Yorkshire and North East Genomic Laboratory.DNA was extracted from 2 to 5\u2009mL lymphocyte blood (EDTA anticoagulant) using Chemagic DNA blood chemistry (CMG-1097-D) on an automated Perkin Elmer Chemagic 360 Magnetic Separation Module and a JANUS Integrator 4-tip Automated Liquid handling platform. DNA was eluted into 400\u2009uL buffer. Extracted DNA samples were measured for DNA yield, concentration and quality using a Nanodrop ND-8000 spectrophotometer. Three MMR genes All women gave written informed consent for tumour and blood testing except deceased cases, whose tumour was obtained and tested with a relative\u2019s consent. Advice from our ethics committee was that the current analysis represented clinical service evaluation and that no specific ethics application was required. There is no directly identifiable patient information presented.2 test.Differences between values were tested by a two-tailed Fisher's \u03c7MLH1 promotor hypermethylation and therefore constitutional LS testing was not performed. Eighteen of the remaining 24 women whose tumours showed MMR deficiency underwent constitutional testing for MMR pathogenic variants. The remaining six did not undergo constitutional analysis because the ovarian cancer case was deceased and a blood lymphocyte sample was not available. An additional 15 women underwent constitutional analysis despite having MMR proficient tumours due to a strong family history, with eight meeting Amsterdam II criteriaIn total, 261 women with ovarian cancer underwent an IHC prescreen for LS . They weMSH2), one MSS (MSH6) and the remaining three were not MSI tested had IHC loss. Only one of the four women<35 years tested for constitutional pathogenic variants had a path_MMR variant identified, and that patient had a parent with four separate bowel primary tumours highly suggestive of LS (MMR pathogenic variants were found in 9/33 (27%) women who underwent constitutional testing with ages of ovarian cancer diagnosis of 33\u201359 years (median 48) . The higve of LS .BRCA1 pathogenic variant. She is unlikely therefore to have developed ovarian cancer via a MMR driven pathway.Here, we describe our 20-year experience of tumour MMR IHC as a prescreen for constitutional testing women with suspected LS-associated ovarian cancer. We tested 261 ovarian tumours for MMR deficiency because women were diagnosed <35 years of age and/or because they had a suggestive personal or family history of LS. Those with strong clinical risk factors underwent constitutional testing even if their tumours were MMR proficient. In total, 27 tumours (10.3%) were MMR deficient and 8 of these had LS. Most were of endometrioid histological subtype. One woman with constitutional path_MMR variant had a MMR proficient tumour; she also had a constitutional et alPrevious studies examining the MMR status of unselected endometrioid or clear cell ovarian cancers found similar rates of MMR deficiency, but overall numbers were very small.MLH-1 promoter methylation status. Analyses were carried out to quality-assured clinical standards in specialist pathology and genetics referral laboratories. Data were collected from our prospective clinical database, ensuring comprehensive reporting of all cases and minimising issues with missing data. All non-deceased women with MMR deficient ovarian tumours unexplained by MLH1 promoter hypermethylation and 15 others, whose clinical risk factors were particularly suggestive, underwent definitive constitutional LS testing using blood lymphocyte DNA. This compares favourably with preceding series where the conversion to constitutional testing was poor and pathogenic variants were assumed from allele frequency in adjacent normal tissue.BRCA1 pathogenic variant and whose tumour is likely to have developed via a non MMR driven pathway.There are several strengths to our work. First, we carried out MMR IHC tumour prescreening for all women referred to the clinical genetics department whose age and family history were suggestive of LS-associated ovarian cancer. We did not restrict testing to any particular histological subtype. This is important because histological subtyping is subjective, challenging in difficult cases and has evolved considerably over the past 20 years, with validated IHC panels increasingly used to assist diagnosis. Many of our cases pre-dated the now gold standard expert gynaecological pathology review and confirmation by IHC.Limitations of the study include failure to conduct MSI analysis for all cases, which precludes a direct comparison between MMR IHC and MSI status as a prescreen for constitutional LS testing. The single centre nature of this study is another limitation, since we cannot necessarily extrapolate our conclusions to other healthcare settings where clinical genetics referral criteria for suspected LS may differ. Our cohort was selected for IHC testing and downstream analyses based on clinical criteria and therefore may not reflect the MMR status of unselected ovarian cancer populations.BRCA1/2 sequencing of women with ovarian cancer to inform suitability for PARP inhibitor therapy and clinical trial enrolment.BRCA2, testing premenopausal women with epithelial ovarian cancer for both BRCA1/2 and LS is appropriate, particularly in an era of panel gene testing where there is little additional cost to add more genes.a priori panel gene somatic testing is unlikely to be indicated.The emergence of targeted therapies has led to mainstream somatic and/or constitutional In summary, we report our experience of MMR IHC as a prescreen for constitutional MMR pathogenic variant testing in women with clinical risk factors for LS-associated ovarian cancer. LS is rare if tumours are MMR proficient. While most LS-associated ovarian tumours are of endometrioid histological subtype, the subjective and sometimes challenging task of pathological interpretation risks misclassification. Thus, our practice is to continue to prescreen all ovarian tumours with clinical risk factors for LS irrespective of tumour histological subtype, especially if their tumour pre-dates recent multidisciplinary panel review in an expert centre."} +{"text": "Chimeric antigen receptor (CAR) T cell immunotherapy uses patient-derived tumor antigen-directed T cells for targeted elimination of cancer cells . The mos18F-FDG PET/CT imaging obtained 3\u00a0months after CAR T cell infusion (A). At the same time, multiple newly enlarged and hypermetabolic cervical lymph nodes were detected in a previously unaffected location (B). These new lesions (red circles and arrows) showed a morphological dedifferentiation with a large central hypodensity compared with nodal DLBCL target lesions at baseline CT (blue circles and arrows). This was also reflected by differences in the radiomic features entropy and uniformity (C). These circumstances triggered a repeat histological workup that determined the transformation of the DLBCL into a sarcoma of the dendritic cells without residual lymphomatous tissue. Based on high PD-L1 expression, checkpoint inhibition with pembrolizumab was initiated.We present a 60-year-old female patient with refractory diffuse large B cell lymphoma (DLBCL) who underwent CAR T cell therapy. During treatment, all lesions decreased in size with a complete metabolic response (Deauville score 1) in Rare cases of transformation into histiocytic and dendritic cell neoplasms have been reported in patients with follicular lymphoma and DLBCL , 5. This"} +{"text": "In the NICU MSSA infections occur frequently and cause morbidity and mortality. Colonization is a risk factor for infection. Optimal infection prevention strategies await a more complete understanding of acquisition and transmission. To investigate possible transmission, we studied whether newly MSSA colonized infants share a strain with another contemporaneously colonized infant.This is a prospective observational study in a level IV NICU from April through November 2019. Infants had weekly MSSA nasal surveillance cultures. Isolates from newly MSSA colonized infants and other infants colonized with MSSA during the same/previous week were subjected to staphylococcal protein A (spa) typing; most pairs with a concordant (CC) spa type were analyzed by whole genome sequencing . Pairs of isolates with a CC spa type and < 25 single nucleotide polymorphism differences on WGS were considered closely related (CC pairs). A control group consisted of pairs of isolates from a newly colonized infant with one randomly chosen colonized infant with a discordant (DC) spa type during the same/previous week. The medical records were reviewed for staff member (SM) and room assignment. Fischer\u2019s exact test was used to compare proportions.Isolates from 60/68 consecutive newly MSSA colonized infants and 111/133 comparison infants were available for spa typing. Of these 60 infants, 23 (38 %) had a CC spa type with another infant colonized during the same/previous week. Of 18 isolate pairs from infants with a CC spa type that were subjected to WGS, 12 (67%) pairs of isolates were closely related. 7/12 (58 %) of CC pairs had a SM in common compared to 2/13 (15 %) in the DC pair groups, p=0.04. 2/12 (17 %) of CC pairs shared a room compared to 2/13 (15 %) pairs in the DC group, p=1.0.Among newly MSSA colonized infants at least 25% are colonized with an isolate closely related to that of another colonized infant indicating likely infant to infant transmission. WGS is more discriminatory than spa typing for MSSA. Given the lack of commonality of room assignment and the commonality of SM assignment, a possible role of healthcare personnel in MSSA transmission should be further investigated.Anne-Catrin Uhlemann, MD, PhD, Merck (Grant/Research Support)"} +{"text": "Community-based organizations (CBOs) are essential settings for older Asian American (AA) and Native Hawaiian Pacific Islander (NHPI) adults for accessing culturally and linguistically appropriate services and connecting with and support each other. This study examined the impact of the COVID-19 pandemic on CBOs\u2019 ability to serve older AA & NHPI adults. This mixed methods study (survey and semi-structured interviews) used a sequential exploratory design. We recruited 65 leaders and staff members from 40 CBOs serving older AA & NHPI adults nationally. Descriptive analysis was conducted with the survey data followed by thematic analysis of the interview data. Many CBOs were impacted by the increased demands for services (80%) and created new services (75%) while experiencing programming disruption (69%), decreased staffing (55%), and loss of revenue (38%). Some CBOs temporarily closed their organizations (38%), while others closed permanently (3%). To remain in operation, many CBOs (65%) increased their online presence, hired staff (52%), and recurred to financial reserves (20%). The semi-structured interviews identified four themes: 1) CBOs resourcefulness to acquire and share resources, 2) technology as a connector for CBOs and an isolator for older adults, 3) heightened racial discrimination against Asians, and 4) emergence of multi-level resilience . CBOs experienced disruption in their operation, and heightened racial discrimination during the pandemic. Yet, CBO\u2019s ability to remain resilient was critical to continue to provide key programs for older adults. Future studies may want to examine evolving needs of CBOs as they adjust to new public health challenges during the pandemic."} +{"text": "Cardiac implantable electronic device (CIED) can interfere with tricuspid valve (TV) function, induce significant tricuspid regurgitation (TR), and worsen patient prognosis . We presSupplemental Video (MP4 2869\u00a0kb)Below is the link to the electronic supplementary material."} +{"text": "COVID-19 has highlighted increasing reliance on information and communication technology (ICT) and challenges in access and use. ICT access also provides resources that benefit users\u2019 mental health. Our study describes changes in the use of ICT before and during the COVID-19 pandemic among cancer patients with and without dementia. We identified 196 (1.6 million weighted population) older adults with a self-reported cancer history who participated in both 2019 and 2020 National Health and Aging Trends Study (NHATS). In 2019, cancer patients with dementia (9.9%) were less likely to use information technology (IT) for health matters compared to those without dementia. In contrast, dementia status was not associated with communication technology (CT) use (email or texts) or IT use for personal tasks (grocery shopping or online banking). IT use for personal tasks was inversely associated with anxiety symptoms and CT use was inversely associated with depressive symptoms (adjusted OR 0.25 (95%CI:0.07-0.97). In 2020, regardless of dementia status, all cancer patients increased their virtual (email/phone/video) contact with family, friends (3.4%-7.0%), and medical providers (17.2%-36.2%) while decreasing in-person contact during the pandemic. This study suggests that there are potential unmet daily needs for patients with comorbid cancer and dementia that may be met with improved ICT access. Such challenges are of increasing concern as COVID-19 has resulted in increased ICT reliance for older adults."} +{"text": "PHY A (phytochrome A) gene. So far, the members of PHY A gene family remains unexplored in Lablab purpureus, and therefore, their functions are still not deciphered. PHYA3 is the homologue of phytochrome A and known to be involved in dominant suppression of flowering under long day conditions by downregulating florigens in Glycine max. The present study is the first effort to identify and characterize any photoreceptor gene in Lablab purpureus and decipher its phylogeny with related legumes.Phytochromes are the best characterized photoreceptors that perceive Red (R)/Far-Red (FR) signals and mediate key developmental responses in plants. It is well established that photoperiodic control of flowering is regulated by PHYA3 was amplified in Lablab purpureus cv GNIB-21 (photo-insensitive and determinate) by utilizing primers designed from GmPHYA3 locus of Glycine max. This study was successful in partially characterizing PHYA3 in Lablab purpureus (LprPHYA3) which is 2 kb longer and belongs to exon 1 region of PHYA3 gene. Phylogenetic analysis of the nucleotide and protein sequences of PHYA genes through MEGA X delineated the conservation and evolution of Lablab purpureus PHYA3 (LprPHYA3) probably from PHYA genes of Vigna unguiculata, Glycine max and Vigna angularis. A conserved basic helix-loop-helix motif bHLH69 was predicted having DNA binding property. Domain analysis of GmPHYA protein and predicted partial protein sequence corresponding to exon-1 of LprPHYA3 revealed the presence of conserved domains (GAF and PAS domains) in Lablab purpureus similar to Glycine max.LprPHYA3 would facilitate the identification of complete gene in Lablab purpureus utilizing sequence information from phylogenetically related species of Fabaceae. This would allow screening of allelic variants for LprPHYA3 locus and their role in photoperiod responsive flowering. The present study could aid in modulating photoperiod responsive flowering in Lablab purpureus and other related legumes in near future through genome editing.Partial characterization of The online version contains supplementary material available at 10.1186/s43141-021-00295-z. Oryza sativa, Lablab purpureus, Phaseolus vulgaris and Glycine max requires photoperiod insensitivity to adapt to a high latitudinal environment is one such underexploited legume with wide range of uses as vegetable, forage, cover crop, split pulse, fodder and medicinal [Lablab purpureus are photoperiod sensitive and indeterminate which flower only during short days, while few improved varieties with determinate growth and photoperiod insensitivity are available which flowers within 40 to 50 days across the year. Photoperiod responsive flowering along with growth habit might have played crucial role in domestication and evolution of this crop. Dominant nature of photoperiod sensitivity, indeterminate growth habit and purple flower was reported along with coupling phase of linkage between photoperiod insensitive flowering and determinate growth habit in Lablab purpureus [Lablab purpureus discerned that photoperiod insensitive flowering and determinate growth habit is linked and they might be governed by recessive alleles of GmPHYA3 and Dt homologs, respectively [TFL governing growth habit has been reported along with involvement of splice site single nucleotide polymorphism (SNP) for growth habit differences in Lablab purpureus [PHYA3 in photoperiod responsive flowering is already reported in Glycine max and Phaseolus vulgaris, and mutations in E3/PHYA3 conferred photoperiod insensitive and early flowering [PHYA3 gene or any marker tightly linked to it in Lablab purpureus has limited the molecular characterization of photoperiod responsive flowering in this crop. Looking to possible role of PHYA3 in photoperiod responsive flowering and lack of genome sequence information, the present work is focused on characterization of this gene in Lablab purpureus using candidate gene approach and phylogenetic analysis of legume phytochromes.The research on photoperiod responsive flowering is mainly anchored to few major pulse legumes edicinal . The moledicinal . Most laurpureus . Moleculectively . Most reurpureus . The rollowering . Lack ofGmPHYA3 sequence from Glycine max [https://www.ncbi.nlm.nih.gov/genbank/). GmPHYA3 is 9.2kb longer, exon 1 was divided into three frames and primers for each frame were designed using web-based Primer BLAST from NCBI . PCR mixture prepared in 200 \u03bcl contained approximately 100 ng genomic DNA, 200 \u03bcM of dNTPs, 10 pmol of forward and reverse primers, standard Taq buffer (Mg2+ plus) and 1 unit of Taq DNA polymerase in total volume of 25 \u03bcl reaction. The PCR cycle involved initial denaturation of 95\u00b0C for 7 min followed by 35 cycles of 94\u00b0C (30 s), 51\u201355\u00b0C (45 s) and 72\u00b0C (1 min) and a final extension of 10 min at 72\u00b0C. Amplicons giving the single discrete band when resolved on 1.5% agarose gel electrophoresis with the expected product size were purified using column purification with SLS PCR Clean-up Kit . Sanger sequencing reaction of purified PCR amplicon was carried out with specific primers using BDT v3.1 Cycle sequencing kit on ABI 3730xl Genetic Analyzer (Applied Biosystems). Bidirectional sequence data obtained from each amplicon were processed using BioEdit [Genomic DNA was isolated from young fresh trifoliate leaves of GNIB-21 using CTAB method . The DNA BioEdit . The bidLprPHYA3 sequence was used as query for BLASTn at https://blast.ncbi.nlm.nih.gov/Blast.cgi for finding homologous sequences with reference to NCBI nucleotide database [LprPHYA3 from Fabaceae family were retrieved from NCBI nucleotide database. MEGA X software [PHYA nucleotide sequences of 16 species from Fabaceae family in addition to Arabidopsis thaliana using the CLUSTAL W alignment algorithm [The processed database . Sequencsoftware was usedlgorithm . All thelgorithm . The inilgorithm . The beshttps://meme-suite.org/meme/) [http://memesuite.org/tools/tomtom), which is the online software performing comparison of given motifs with available databases. The output generated from TOMTOM include sequence-logo graphics on behalf of the alignment of two motifs with p and q value of the match [The detection of conserved motifs in DNA sequence of phytochrome gene was performed using online tool MEME (Multiple Em for Motif Elicitation) (g/meme/) . This onhe match .LprPHYA3 was subjected to exon prediction using Eukaryotic GeneMark.hmm version 3.54 [GmPHYA3 sequence. The protein sequence of exon 1 from Lablab purpureus GNIB-21 was used to perform BLASTp in NCBI GenBank database and the sequences showing homology were further used to create multiple alignment using the CLUSTAL W algorithm [PHYA in 21 different plant species was also performed using JTT model in MEGA X [http://smart.embl-heidelberg.de/) Version 9 was used for predicting domains in both GmPHYA3 and LprPHYA3 protein sequences [The identified partial sequence of ion 3.54 . The nuclgorithm , 31. Phyn MEGA X , 37. Theequences .Lablab purpureus cv GNIB-21, which is determinate and photo-insensitive, was used for characterizing PHYA3 gene. The sequencing data after processing and analysis revealed the successful characterization of the exon-1 of PHYA3 gene in Indian bean (LprPHYA3- Lablab purpureus PHYA3) for the first time in the world. BLASTn analysis of nucleotide sequence LprPHYA3 indicated highest identity with Vigna unguiculata (95.76%) followed by Vigna angularis (95.67%) and Glycine max (90.90%), all with E-value close to zero.In the present study, LprPHYA3 nucleotide sequence was compared with the nucleotide sequences of phytochrome genes of other plant species, most of which, belonged to the Fabaceae family. Phylogenetic analysis showed that this phytochrome A gene evolved from common ancestry root but diverged into different clades during the course of evolution , two PAS (period\u2013ARNT\u2013single-minded) domains and two Histidine kinase-related domains (HKRD) viz., HisKA and HATPase_c as depicted in Fig LprPHYA3 showed two domains, i.e. GAF sand PAS is considered a strong candidate for FT3 and mutations in GmPHYA3 results in early flowering [E3 have been reported in Glycine max and Phaseolus vulgaris that confer significantly early flowering and photoperiod insensitivity [E3 mutants depicted a large insertion in fourth intron and one SNP for non-synonymous amino acid substitution in third exon. Naturally occurring e3 allele carries a large deletion spanning exon 4, whereas an induced mutant e3 allele has sustained a 40-bp deletion and frameshift in the middle of exon 1 [E loci can be used for more efficient breeding strategies [Lablab purpureus, photoperiodic response of flowering may also be governed by circadian clock as it is reported in Glycine max. CONSTANS (CO) is a circadian-regulated gene and acts as prime regulator of this pathway. It activates the expression of florigen gene FT by coordinating light and clock input to leaves. The different genes, their responses to photoperiod and their putative roles mediating this process have been proposed. It is speculated that under SD condition (low R:FR), E3 and E4 are repressed due to lack of exorbitant light condition and thus E1 is also suppressed which thus has no effect on FT2a and FT5a genes leading to early flowering phenotype. Under enriched light condition, i.e. LD condition PHYA3 (E3) and PHYA2 (E4) are expressed activating the expression of E1 which eventually leads to FT2a/FT5a downregulation, flowering repression, indeterminate growth (if TFL1 is present) and delayed maturity. The presence of TFL1 and/or tfl1 convoys indeterminate and determinate growth habit, respectively [TFL allele suppresses development of floral architecture at shoot apex in indeterminate types; racemes emerge from axillary bud only upon short day conditions where FTs, expressed due to favourable photoperiod condition, might be able to nullify the effect of TFL in competitive manner. Determinate growth habit results from non-functional allele (tfl) of TFL which is unable to suppress flowering in shoot apex and results into determinate growth habit and photoperiod insensitive flowering. This is unveiled by the presence of splice site SNP at third exon in Lprtfl which renders non-functional protein and is responsible for determinate growth habit and photo-insensitive flowering in Lablab purpureus cv GNIB-21 [TFL1 locus in cowpea also resulted in determinate growth habit [The deciphering of putative molecular pathways in legumes involving different phytochrome genes and their involvement in governing flowering time will pave way for future research. Various genetic models for this regulatory framework of photoperiod-based flowering with different known loci is available for sitivity . Additiorategies . These pectively . Dominan GNIB-21 . A non-sth habit .Fig. 5A A genes in some legumes along with their putative role is represented in photoperiod adaptation of short-day legumes like Glycine max and Phaseolus vulgaris, it becomes quite important to understand its role in Lablab purpureus which is also a short-day crop. Due to the lack of genome sequence database, there is very scanty information available about Lablab purpureus at molecular level. The present study was successful in characterizing exon-1 in LprPHYA3 by utilizing GmPHYA3 sequence as reference. Comparative gene mapping has been quite useful in deducing genomic structure from related plant species due to conservation of genetic content, gene order and function [PHYA3 gene has most probably been evolved from common ancestral PHYA gene of these species, as depicted from the tree. Previous studies have also shown closeness of Lablab purpureus to Vigna unguiculata and their predictable evolution from Glycine max in Phaseoleae clade [PHYA3 among the phaseoleae clade legumes indicating evolutionary closeness with Vigna unguiculata, Vigna angularis and Glycine max employing both DNA and protein sequences for phylogenetic analysis comprising N-terminal extension (NTE), period\u2013ARNT\u2013single-minded (PAS), cGMP-specific phosphodiesterase\u2013adenylyl cyclase\u2013FhlA (GAF) and phytochrome-specific (PHY) domains with the help of a bilin chromophore. C-terminal output module (OPM) is shared by two PAS on the N-terminal side and a histidine kinase-related domain (HKRD) [LprPHYA3-Exon 1 encodes GAF and PAS domain of phytochrome genes which belongs to the photo-sensory module and is responsible for convertible Pr/Pfr transformation as well as light-signal transduction from this module to output module, respectively [PHYA3 in Lablab purpureus would unravel the different domains involved in downstreaming light-mediated response to the signaling pathway along with their putative roles.Phytochromes mediates light responses by interacting with multiple partners to modulate transcription of downstream target genes. The transcription factor (TF) containing basic helix-loop-helix (bHLH) motif interacts physically with red and far-red photoreceptor, phytochrome, called Phytochrome Interacting Factors (PIF) . The preene Fig. . The TF bidopsis , 53. Prelowering . The preYA3 Fig. which arn (HKRD) . NTE is n (HKRD) . LprPHYAectively . CharactLprPHYA3 would facilitate allelic characterization in relation to photoperiod responsive flowering in Lablab purpureus. Phylogenetic analysis indicated that complete characterization of LprPHYA3 would be possible utilizing sequence information from related legumes. The presence of conserved DNA binding motif (bHLH69) in PHYA gene indicated that it might repress flowering by competing for DNA binding with bHLH containing TFs. Domain analysis of protein-encoding LprPHYA3 would unfold the signaling pathways and their interaction with different proteins from PEBP (Phosphatidyl ethanolamine-binding protein) family genes that would guide flowering response.Partial characterization of LprPHYA3 and their role in modulating photoperiod responsive flowering. Additionally, qPCR studies could also be undertaken for relative expression studies of PHYA3 in LD and SD conditions. The role of LprPHYA3 may be confirmed through genome editing by utilizing partial sequence reported in the present study. These efforts would accelerate the understanding of flowering time and growth habit regulation in Lablab purpureus in response to changed photoperiod.The continued progress in this direction would entice further questions to address in future like characterization and identification of allelic variants for Additional file 1. Sequencing data."} +{"text": "Neurotransmitter sodium symporters (NSS) are a subfamily of SLC6 transporters responsible for regulating neurotransmitter signalling. They are a major target for psychoactive substances including antidepressants and drugs of abuse, prompting substantial research into their modulation and structure-function dynamics. Recently, a series of allosteric transport inhibitors have been identified, which may reduce side effect profiles, compared to orthosteric inhibitors. Allosteric inhibitors are also likely to provide different clearance kinetics compared to competitive inhibitors and potentially better clinical outcomes. Crystal structures and homology models have identified several allosteric modulatory sites on NSS including the vestibule allosteric site (VAS), lipid allosteric site (LAS) and cholesterol binding site (CHOL1). Whilst the architecture of eukaryotic NSS is generally well conserved there are differences in regions that form the VAS, LAS, and CHOL1. Here, we describe ligand-protein interactions that stabilize binding in each allosteric site and explore how differences between transporters could be exploited to generate NSS specific compounds with an emphasis on GlyT2 modulation. Neurotransmitter sodium symporters (NSS) are secondary active transporters that regulate synaptic concentrations of neurotransmitters via reuptake into surrounding glial cells or presynaptic terminals. Members of the solute carrier 6 (SLC6) family act on a broad range of neurotransmitter substrates: glycine (GlyTs), dopamine (DAT), serotonin (SERT), noradrenaline (NET) and \u03b3-aminobutyric acid . ImpaireAquifex aeolicus bacterial leucine transporter (LeuT) and supplemented by recently solved structures of the eukaryotic Drosophila melanogaster dopamine transporter (dDAT), human serotonin (hSERT) and human glycine transporter type 1 (hGlyT1) . Substra(hGlyT1) . Crystal(hGlyT1) . Current(hGlyT1) . This isThe structure and function of SLC6 neurotransmitter transporters has been extensively reviewed elsewhere and thusThe substrate site (S1) of the SLC6 family is located approximately halfway across the membrane and is formed by TMs 3 and 8 and the unwound regions of TMs 1 and 6 . This hyThere has been debate over whether there is a second substrate site (S2) located in the extracellular-facing vestibule above the S1 site. In LeuT, dissociation of sodium from the second sodium site (Na2) allosterically modulates the transporter, allowing cytoplasmic release of leucine from S1 . It has + and Cl\u2212. SERT, DAT, and GlyT1 couple the transport of substrate with the co-transport of 2Na+ and 1Cl\u2212 that mediates this interaction. Mutating this residue in GAT1 (S331D/E), GAT4 (S340E), DAT (S375E), and GlyT1 (S339D/E) results in a loss of chloride dependence that is analgesic in rodent models of neuropathic pain and superfusion of lamina II neurons in the dorsal horn of rat spinal cord slices delays the decay of glycinergic dependent currents without altering their amplitude (50 = 8\u00a0\u03bcM) but achieves complete inhibition compared to the partial inhibition observed with NAGly (50= 11.9\u00a0\u03bcM) but maintains a similar level of inhibition as NAGly (50 = 340\u00a0nM) has a C18 monounsaturated acyl tail and is the most potent acylcarnitine whilst di-unsaturated or fully saturated tails do not generate substantial inhibition (50 = 880\u00a0nM) compared to NAGly but is not as potent as OLCarn (N-arachidonyl glycine (NAGly) is an endogenous lipid with a 20-carbon (C20) polyunsaturated tail conjugated to a glycine headgroup. NAGly is most concentrated in the spinal cord and has been proposed to regulate nociceptive pathways with studies showing that intrathecal administration is analgesic in rodent neuropathic and inflammatory pain models . NAGly implitude . Togetheth NAGly . N-arachas NAGly . Differias NAGly . Screenihibition . Similarhibition . The infs OLCarn . Thus, bcis configuration are active and lipids with double bonds in the trans configuration are inactive was used for in vivo experiments and found to be analgesic in a rat model of chronic neuropathic pain, highlighting the promise of GlyT2 as a therapeutic target for analgesia that modulate transport activity have been identified in NSS family members . High se"} +{"text": "This survey study examines association of housing insecurity with psychological distress and self-rated health among US adults during the COVID-19 pandemic. Understanding the association between housing insecurity and health in a nationally representative sample during the pandemic is critical to inform efforts to support people harmed by the economic downturn.Economic hardship due to the COVID-19 pandemic has exacerbated concerns about the threat of evictions and foreclosures.AAPOR) reporting guideline and the Survey Reporting Guideline (SURGE).This survey study was deemed nonhuman participants research by the institutional review board at Johns Hopkins Bloomberg School of Public Health, and informed consent was waived. This study followed the American Association for Public Opinion Research , which had a response rate of 70.4%. Among wave 1 respondents, 1337 responded to wave 2 (July 2020), and 1222 responded to wave 3 with completion rates of 91% and 92%, respectively.Data were from wave 3 of the Johns Hopkins COVID-19 Civic Life and Public Health Survey5 and dichotomized as moderate or severe vs lower psychological distress based on a score of 5 or more and self-rated health dichotomized as fair or poor vs good, very good, or excellent.Housing insecurity was defined as being currently behind on rent or mortgage or having no or low confidence in their ability to pay the next rent or mortgage payment vs having moderate or high confidence. Outcomes included psychological distress symptoms measured using Kessler 6 Psychological Distress Scale6 included gender, race and ethnicity, age, household income in 2019, employment status, housing tenure, household structure, and urbanicity ; non-Hispanic Black race (44 [37%] vs 113 [8%]), were aged 30 to 54 years (88 [64%] vs 472 [38%]), earned less than $35\u2009000 in 2019 (64 [52%] vs 268 [27%]), lived with children (58 [47%] vs 279 [29%]), or resided in metropolitan counties (116 [92%] vs 914 [84%]) . Overall, 128 participants (12%) reported housing insecurity in November 2020. Among participants experiencing housing insecurity, 42 participants (34%) reported being behind on housing payments, 55 participants (38%) reported having little to no confidence in their ability to make the next housing payment, and 31 participants (28%) reported both . Housing4 [84%]) .P\u2009=\u2009.09), and 26% of participants experiencing housing insecurity reported fair or poor health, relative to 15% of participants with housing security (P\u2009=\u2009.03) of participants reported severe to moderate psychological distress; 18% reported fair to poor health. Compared with participants with housing security, participants experiencing housing insecurity reported higher distress (69 [57%] vs 467 [45%]) and lower self-rated health (33 [30%] vs 157 [16%]). After covariate adjustment, housing insecurity and lower self-rated health was statistically significant. The association between housing insecurity and higher distress was no longer statistically significant. Fifty-seven percent of participants experiencing housing insecurity reported moderate or severe distress relative to 45% of participants with housing security (P\u2009=\u2009.03) .In this nationally representative sample of US adults, housing insecurity was associated with higher psychological distress and lower self-rated health during the COVID-19 pandemic. The survey was conducted after the United States Centers for Disease Control and Prevention\u2019s nationwide eviction moratorium, which may have attenuated these associations. Interventions that reduce housing insecurity during the COVID-19 pandemic will promote the health of the US population, and those interventions that consider equity in their implementation may mitigate entrenched health disparities resulting from structural racism and exacerbated by the pandemic. Results may be vulnerable to sampling biases, including the underrepresentation of adults experiencing homelessness. The cross-sectional design of this study precludes identifying causality; the survey did not assess prepandemic housing needs."} +{"text": "Titanium(III) isa useful strong reductant and is usually standardizedwith iron(III) in volumetric analysis. Iron(III) is widely used asan oxidant and is usually standardized with thiosulfate ions throughan iodine liberation reaction. The evaluation of the standardizationprocedure for iron(III) with thiosulfate ions is therefore essentialto ensure the reliability of standardized titanium(III) solutions.To investigate the titration procedure for iron(III), two differenttitrations were performed: redox titration with thiosulfate ions throughan iodine liberation reaction and chelatometric titration with disodiumdihydrogen ethylenediaminetetraacetate. Subsequently, for the investigationof standardization of iron(III), titanium(III) was assayed throughtwo titration paths: redox titration with standardized iron(III) andredox titration with standard potassium dichromate. The reliabilityof titrimetric procedures was evaluated by applying several differentstoichiometric reactions to each chemical. All titrimetric procedureswere consistent with each other within their expanded uncertaintiesand were capable of providing reliable volumetric standards with carefuloperations presented in this study. Titanium(III)is often standardized with iron(III).4 Iron(III)is usually standardized with thiosulfate through an iodine liberationreaction.6 Thiosulfate is usually standardized witha certified reference material (CRM) of iodate or dichromate. Thesetitrimetric methods play a key role in the accuracy of the titrationresults.7 Useful metrological informationon the reliability of the volumetric standards such as these veryweak oxidant and very strong reductant has not been found. The aimof the present study is to evaluate the reliability of the titrimetricprocedures for iron(III) and titanium(III) as a volumetric standard.A reliable titanium(III) standard solutionbecame necessary whenthe authors were trying to determine the purity of the perchloratesalt.5 A standardization procedure for iron(III) isas follows: iodine (triiodide ions) liberated by iron(III) in an acidicpotassium iodide solution (Ammonium iron(III) sulfate dodecahydrate is often used as a sourcematerial for the reagent solution or the standard solution of iron(III).solution 1 is titrsolution 2 and 3; solution 2 and 3:113 The accuracy of the titrimetric procedure for iron(III) would belower than that for these strong oxidants because the lower oxidizingability of iron(III) would lead to a smaller oxidation rate of iodideions. Longer experimental time would lead to larger biases due toside reactions such as the oxidation of iodide ions by atmosphericoxygen and the volatilization of generated iodine.The iodineliberation process is significantly affected by theamounts of acids and potassium iodide, the waiting time for liberation,and light. The process, therefore, plays a key role in the accuracyof standardization of iron(III). One of the authors has discussedseveral appropriate standardization procedures through the iodineliberation reaction for strong oxidants such as iodate, cerium(IV),bromate, periodate, dichromate, and osmium(VIII).In the presentstudy, iron(III) was assayed through two titrationpaths to investigate the iodine liberation reaction 1: redox 2(SO4)3] or titanium(III) chloride [TiCl3] are often used asa source material of titanium(III). The sulfate salt was chosen asa source material of titanium(III) in this study to avoid the oxidationof chloride ions in redox titrations. Titanium(III) was assayed throughtwo titration paths to examine the accuracy of titration procedures(15Titanium(III) sulfate on each surface.21 The current was proportionalto a certain amount of liberated iodine.Constant voltage biamperometry was utilizedto roughly investigatethe iodine liberation profile. Biamperometry is a method for monitoringthe current between typically twin platinum electrodes where a constantvoltage is applied (500 mV). The current flows in the presence ofboth iodide ions and triiodide ions through the reaction [3I\u20131 hydrochloric acid were added to the solution. The solution was titratedwith a sodium thiosulfate solution at 0 to 50 min after the iodineliberation started. A stopper beaker was used during the liberationto prevent the vaporization of iodine. The concentration of the sodiumthiosulfate solution was standardized with standard potassium iodatein advance. The end point was detected by constant voltage biamperometry.The indicator current proportionally decreased with the titrationproceeding near the end point and the current was kept at zero amperesafter the end point. The end point was the intersection between thelinear regression curve of indicator currents versus the amounts of the sodium thiosulfate solution used and the baseline(0 \u03bcA).22The purity of ammoniumiron(III) sulfate dodecahydrate as Fe throughthe iodine liberation reaction was examined by gravimetric titration.Approximately 1.8 g of ammonium iron(III) sulfate dodecahydrate wasplaced in a 50 mL beaker and dissolved in 50 mL of pure water. Approximately3 g of potassium iodide and 0 to 13.2 mL of 6 mol L\u20131 solution of titanium(III)sulfate was prepared by diluting a titanium(III) sulfate solution(20% in 1 to 4% sulfuric acid) two times with 2 mol L\u20131 sulfuric acid. The solution was kept in a HDPE bottle of Nalgene.An approximately 250 mol kg\u20131 sulfuric acid. The potassium dichromate solution was gravimetricallytitrated with the titanium(III) solution on heating around 90 \u00b0C.The end point was detected by potentiometry with a Pt\u2013Ag/AgClcombination electrode. The inflection point calculated by third-orderpolynomial approximation was decided as the end point.22Approximately 0.18 g of standard potassium dichromate was placedin a 50 mL beaker and dissolved with 20 mL of 2 mol L\u20131 sulfuric acid. The iron(III) solution was gravimetricallytitrated with the titanium(III) solution on heating around 50 \u00b0C.The end point was detected in the same manner mentioned above.Approximately 1.6 g of ammonium iron(III)sulfate dodecahydratewas placed in a 50 mL beaker and dissolved with 20 mL of 2 mol L"} +{"text": "MSH2 mutations in LS are still lacking. The aim of this study was to comprehensively analyze the clinicopathological characteristics and molecular basis of colorectal cancer (CRC) in patients with uncommon MSH2 cytoplasmic expression.A large proportion of patients with Lynch syndrome (LS) have MSH2 abnormalities, but genotype-phenotype studies of We retrospectively reviewed 4195 consecutive cases of CRC patients diagnosed between January 2015 and December 2017 at the Cancer Hospital Chinese Academy of Medical Sciences. Of the 4195 patients with CRC, 69 were indicated to have abnormal MSH2 expression through tumor immunohistochemical staining. Genetic tests, such as next-generation sequencing, large genomic rearrangement (LGR) analysis, microsatellite instability status analysis and genomic breakpoint analysis, were performed. Clinicopathological and molecular characteristics and clinical immunotherapy response were analyzed.MSH2 and/or EPCAM. Of these LS patients, 26.7% were confirmed to harbor large genomic rearrangements (LGRs). Of note, three tumors from two unrelated family pedigrees exhibited a rare cytoplasmic MSH2 staining pattern that was found in LS patients with EPCAM/MSH2 deletions. RNA analysis showed that two novel mRNA fusions of EPCAM and MSH2 resulted in the predicted protein fusion with MSH2 cytoplasmic localization. Analyses of genomic breakpoints indicated that two novel deletions of EPCAM and MSH2 originated from Alu repeat-mediated recombination events. Our study also provides clinical evidence for the beneficial effect of the PD-1 inhibitor pembrolizumab for CRC patients that exhibit cytoplasmic MSH2 staining.Forty-five of 69 patients were identified to have LS with pathogenic germline mutations in EPCAM and\u00a0MSH2.Our study demonstrates that the rare cytoplasmic MSH2 staining pattern should be fully recognized by pathologists and geneticists. Given the specific genotype-phenotype correlation in LS screening, we advocate that all CRC patients with cytoplasmic MSH2 staining in histology should be screened for LGRs of MLH1, MSH2, MSH6 and PMS2) and deletions in EPCAM , an autosomal dominant hereditary disorder, is the most common colorectal cancer (CRC) predisposition syndrome, accounting for 1%\u20133% of all newly diagnosed CRCs in the Cancer Hospital of the Chinese Academy of Medical Sciences between January 2015 and December 2017, we identified a cohort of 69 patients with loss of the MSH2 and/or MSH6 proteins who had been screened by IHC staining for tumor MMR proteins. Detailed information on the CRISPLS cohort was previously reported , PMS2 (clone EPR3947), MSH2 (clone FE11), MSH6 (clone EP49) , and BRAF V600E (VE1) were used. Briefly, after deparaffinization, rehydration and antigen-retrieval, 4-\u03bcm-thick sections were stained in a Ventana Benchmark IHC automated slide stainer and visualized using the OptiView DAB IHC detection kit . The absence of nuclear staining in tumor cells or very faint nuclear staining in focal tumor cells was defined as loss of protein expression . Stromal/lymphoid cells and nearby normal glandular epithelium of the bowel served as positive internal controls.Microsatellite instability (MSI) testing was performed on tumor and normal DNA using a fluorescence PCR-based assay in which six mononucleotide repeat markers and two pentanucleotide repeat loci (Penta-C and Penta-D) were amplified to confirm the identity of paired tumor and benign tissues. The PCR products were run on an Applied Biosystems 3500 Genetic Analyzer and analyzed using GeneMapper v5.0 software . Tumors with shifts in two or more markers were classified as unstable MSI-high , 24.Formalin-fixed, paraffin-embedded tumors and adjacent normal tissue were collected from the cohort. Genomic DNA was extracted using a TGuide Genomic DNA One-Step Kit and a TGuide Automated Nucleic Acid Preparation Instrument according to the manufacturer\u2019s instructions as previously described .MMR genes from genomic DNA extracted from normal FFPE samples according to the manufacturer\u2019s instructions. Molecular barcoded DNA libraries were hybridized with a commercial ClearSeq Inherited Disease multigene panel that covered total exons and intron boundaries within at least \u00b120 bases of the EPCAM, MLH1, PMS2, MSH2 and MSH6 genes (Agilent Technologies). A detailed protocol for variant annotation and classification was described previously for germline mutation testing of eviously , 25. DiaMSH2 and EPCAM genes among MSH2-deficient patients with no germline mutations identified by next-generation sequencing were assessed by MLPA using the SALSA MLPA P003 MLH1/MSH2 kit (including the 3\u2019 end of EPCAM) and P072 MSH6 kit (including the EPCAM/MSH2 region) . Fragment analysis of amplified genomic DNA extracted from normal FFPE samples was performed on an ABI3500 capillary sequencer (Applied Biosystems). The MLPA data were quantitatively analyzed using Coffalyser.Net software (www.mlpa.com).Large genomic rearrangements (LGRs) in st Strand cDNA Synthesis Kit and analyzed for EPCAM-MSH2 fusion transcripts. Polymerase chain reaction (PCR) products were loaded directly on 2% agarose gels and visualized under UV illumination. Selected PCR products were sequenced on an ABI 3500xl capillary DNA analyzer . Details of the PCR primers used are provided in Total RNA was extracted from the peripheral blood leukocytes of patients using TRIzol reagent (Agilent Technologies). cDNA was synthesized using a PrimeScript II 1EPCAM and MSH2 genes using a TaKaRa LA PCR Kit according to the manufacturer\u2019s protocol. The PCR products were analyzed by electrophoresis on ethidium bromide-stained 1% agarose gels and then subjected to UV detection. The expected fragment was purified and sequenced on an ABI 3500xl capillary DNA analyzer (Applied Biosystems). Details of the PCR primers are provided in A series of long-range PCR experiments designed to span the putative deletion region were performed to characterize the exact breakpoints in the A univariate analysis of categorical variables was performed by cross tabulation using a chi-square test to compute p-values. An unpaired t test was used for continuous variables. Statistical descriptions or analyses were conducted using SPSS or Prism software. All tests were 2-tailed, and p-values < 0.05 were considered statistically significant.We retrospectively enrolled a consecutive cohort of 4195 CRC patients. Among these patients, 345 were eligible, exhibiting dMMR, and 69 exhibited abnormal MSH2 protein expression were confirmed to carry LGRs in MSH2/EPCAM by MLPA, including six probands with MSH2 genomic deletions, four with MSH2-EPCAM deletions (two cases from a family pedigree), and two with EPCAM deletions. The clinicopathological and molecular findings for these 12 patients with LGRs are presented in MSH2/EPCAM LGRs exhibited an earlier age of CRC onset (mean: 43.8 years) than those with LS with MSH2 SNV/indel (mean: 49.9 years).To estimate the frequency and specificity of us study . GermlinMSH2 and EPCAM in all three patients. Two patients had a heterozygous large genomic deletion in EPCAM (exons 3\u20139) and MSH2 (exon 1). One patient harbored heterozygous deletion of exons 3\u20139 of EPCAM exhibited rare cytoplasmic MSH2 localization in tumor cells but showed patchy expression of the MSH6 protein that was somewhat weaker in tumor cells than in internal control cells (patient 164 and patient 271 from one family pedigree and patient 345) S2. TheyMSH2/EPCAM LGRs (http://insight-database.org). Repeated Alu elements are implicated in the etiology of genomic rearrangement for many inherited cancers (http://www.repeatmasker.org) revealed that these breakpoints lay within Alu elements that share high sequence identities. A schematic diagram is shown in To elucidate the molecular characteristics of cytoplasmic localization of MSH2 in tumors, we performed mapping analysis of gene fusions and gene breakpoints for two individual patients. Sanger sequencing of cDNA from the blood lymphocytes of the patients suggested that a fusion transcript of EPCAM and MSH2 was the source of aberrant MSH2 localization in tumors. Sequencing revealed a common fusion of exon 1 of EPCAM and exon 2 of MSH2 in patient 271 and patient 345. On the basis of this fusion transcript analysis, we performed a series of long-range PCR experiments on the region of interest between intron 1 of EPCAM and intron 1 of MSH2 to identify the precise breakpoint in the CAM LGRs S7. Ulti cancers . Our anaEPCAM and MSH2 genes. Patient 271 was treated with the PD-1 inhibitor pembrolizumab in combination with capecitabine every 3 weeks for 19 cycles. The patient achieved a clinical partial response (PR) with a 56% reduction in the short axis diameter of the enlarged retroperitoneal lymph node, as revealed by computed tomography (CT) scans after a 19-month course of treatment was diagnosed with retroperitoneal lymph node metastasis of stage IV colon cancer with rare MSH2 cytoplasmic localization. This patient underwent dissection of distal colon cancer at the age of 33.\u00a0A cancer family history survey for this patient showed that nine members in three consecutive generations suffered from LS-related cancers (LSRC), including CRC in II6, III2, III5, III7, III11, III13, IV1; pancreatic cancer in II6; and endometrial cancer in III10 LS patients were carriers of MSH2 and/or EPCAM LGRs. Three of 12 (25%) probands with MSH2/EPCAM LGRs harbored a rare MSH2 chimeric fusion protein that was detectable in the cytoplasm of tumor cells by IHC. We also provided clinical evidence that a CRC patient harboring cytoplasmic MSH2 fusion proteins was responsive to treatment with immune checkpoint inhibitors.In recent years, there has been increasing demand for immunotherapy and LS screening among CRC patients displaying MSI-high or dMMR \u201330. Thesquencing , 31. In MSH2 is a frequent causal event among LS patients staining method , transmembrane domain and cytoplasmic domain . Patients with cytoplasmic staining have a common feature of MSH2 C-terminal fusion with an EPCAM N-terminal fragment of 25 amino acids, including a complete signal peptide . The sigMSH2/EPCAM LGRs, consistent with previous studies , Beijing Hope Run Special Fund of the Cancer Foundation of China (LC2017B14), the Non-Profit Central Research Institute Fund of Chinese Academy of Medical Sciences (2019PT310026) and the National Key Research and Development Program (2017YFC1311005). The funders had no role in the study design, data acquisition, analysis, interpretation, writing or submission of the manuscript.Author YZ was employed by company Beijing Microread Genetics.The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Median hospital stay for both groups was 1.0 day. The pregabalin group had significantly lower consumption of total opioids during admission compared to the control group . Although pain scores in the treatment group (3.21 \u00b1 2.03) were lower than in the control group (3.71 \u00b1 2.95), the difference was not statistically significant . DISCUSSION/SIGNIFICANCE OF IMPACT: Based on the results, a one-time preoperative oral dose of pregabalin before orthognathic surgery in patients with cleft lip and palate reduced total opioid consumption during admission. However, there was no difference in length of stay or pain scores within the two groups. A single preemptive oral dose of pregabalin should be considered an effective adjunct to pain management protocols in patients undergoing orthognathic surgery.OBJECTIVES/GOALS: The current opioid epidemic has placed post-operative pain management under scrutiny. Limiting post-operative pain can decrease overall opioid usage in the recovery period, especially after orthognathic surgery. Several studies have illustrated the efficacy of pregabalin in decreasing postoperative pain and opioid usage in adults undergoing orthognathic surgery. We aim to study the effects of a single dose of preoperative pregabalin on postoperative pain and total opioid consumption after orthognathic surgery in individuals with cleft lip and palate. METHODS/STUDY POPULATION: This was a retrospective cohort study of consecutive patients who received Le Fort I midface advancement between June 2012 and July 2019 by one of two surgeons at a single institution. We took advantage of our institution\u2019s implementation, beginning in 2016, of a one-time dose of preoperative pregabalin for LeFort I midface advancement. All patients had diagnosed cleft lip and palate. The treatment group received a one-time preoperative dose of pregabalin. The control group did not receive pregabalin. Total morphine milligram equivalents (MME) consumption was calculated by adding intraoperative opioid administration and postoperative opioid consumption during admission. Postoperative pain control during admission consisted of oral oxycodone and intravenous (IV) hydromorphone or morphine. Duration of hospitalization and pain intensity assessed with the numeric pain rating scale (0-10) were also recorded. The mean postoperative pain assessment scores during admission was calculated for each patient. The median of these individual mean pain assessment scores for each group was subsequently computed. RESULTS/ANTICIPATED RESULTS: Twenty-three patients were included in this study; 12 patients received pregabalin . Mean age (years) at operation of the pregabalin (18.3 \u00b1 1.9) and control groups (17.8 \u00b1 1.9) were also equivalent ("} +{"text": "In theirgroup of 28 patients with RA, dyspnoea was present in 47%, with ILDdemonstrated on chest radiograph in 18%. Abnormal spirometry waspresent in 25%. Unfortunately, chest computed tomography (CT) wasnot available. These figures are not too dissimilar to the findings ofMorrison et al.,[3] who investigated patients with RA in Cape Town inthe early 1990s, prior to the advent of chest CT.The prevalence of interstitial lung disease (ILD) in rheumatoidarthritis (RA) varies between 7.7 and 67%, depending upon thediagnostic methods available.[4] Traditional diagnostic methods such as lungbiopsy can often be avoided if typical HRCT patterns are present.Other diagnostic methods include pulmonary function testing, whichis nonspecific for diagnosis but is extremely useful in prognostication.[5]Respiratory symptoms and clinical signs in RA are common butnonspecific. The advent of high-resolution CT (HRCT) has allowedearlier and more precise detection of RA-ILD. The most commonpatterns identified are usual (UIP) and nonspecific interstitialpneumonitis (NSIP).et al.[6] demonstrated pleural nodulesor B-lines in 28% of patients with RA studied, HRCT confirmedthe presence of early ILD. A further study comparing standard and\u2018pocket\u2019 ultrasound devices showed sensitivities of 92% and 89%,with specificities of 56% and 50%, respectively, when compared withHRCT.[7]Ultrasound of the lung is now widely used, with ultrasoundequipment becoming increasingly available and portable at relativelylow cost. Lung ultrasound has been investigated as a screening testfor RA-ILD. Moazedi-Fuerst RA is ubiquitous and many areas of the world do not have readyaccess to sophisticated radiological techniques such as HRCT.Treatment of RA-ILD requires suspicion and confirmation of diagnosis before introduction of potentially harmful agents such ashigh-dose corticosteroids, immunosuppressive or antifibrotic therapy.Ultrasound should be investigated further as an accessible tool for usein resource-limited environments."} +{"text": "This article examines the budgetary implications of high-priced accelerated approval drugs by estimating Medicare spending on these drugs from 2015 through 2019. Drugs granted accelerated approval must undergo confirmatory studies, but in some cases these studies have not been completed in a timely fashion,4 We obtained annual Medicare Part B and D spending between 2015 and 2019 from publicly available Drug Spending Dashboards. For Medicare Part D, we estimated net spending by subtracting estimated average annual rebates and other discounts for specific drug classes based on data from a recent federal report.5 In a sensitivity analysis, we alternatively used estimated drug-specific rebates from SSR Health. Part B spending data already reflect discounts, but we adjusted spending to account for missing data for 7 newly marketed drugs reporting guidelines for economic analyses and was not submitted for institutional review board approval because it used public, nonidentifiable data (45 CFR 46.102).We stratified spending based on periods when drugs were exclusively approved via the accelerated approval pathway vs periods when drugs were approved for multiple indications via both the accelerated and traditional approval pathways, prorating annual spending estimates as necessary. We compared spending on accelerated approval drugs with total estimated net Medicare Part B and D spending.A total of 66 drugs with Medicare spending from 2015 through 2019 had at least 1 accelerated approval indication. Among these, 49 (74%) were oncologic drugs. Annual spending on all drugs with an accelerated approval indication increased from 2015 to 2019 , but spending on drugs with only accelerated approval indications decreased , pembrolizumab ($6.1 billion), and bevacizumab ($3.4 billion); the highest-spending Part D drugs were ibrutinib ($6.6 billion), palbociclib ($1.6 billion), and droxidopa .Only 13% of Medicare spending on accelerated approval drugs occurred while these drugs were exclusively marketed under the accelerated approval pathway, while most spending was on drugs with multiple indications approved via both accelerated and traditional FDA pathways. This poses a challenge for policy makers seeking to lower Medicare spending on drugs with unproven benefit, since Medicare neither reports indication-specific drug spending nor reimburses drugs differently based on patients\u2019 diagnoses. While accelerated approval drugs are approved based on pivotal trials assessing surrogate measures, some may offer more benefits to patients than others. If the goal of policy makers is to better align Medicare drug spending with clinical value, focusing on accelerated approval drugs alone will be insufficient."} +{"text": "Leishmania donovani centrin deleted parasite vaccine (LdCen-/-). However, neutrophil-DC interactions in T cell priming in vaccine immunity in general are not known. In this study we evaluated the interaction between neutrophils and DCs during -/-LdCen infection and compared with wild type parasite (LdWT) both in vitro and in vivo.Neutrophils are involved in the initial host responses to pathogens. Neutrophils can activate T cell responses either independently or through indirect involvement of Dendritic cells (DCs). Recently we have demonstrated direct neutrophil-T cell interactions that initiate adaptive immune responses following immunization with live attenuated -/-LdCen parasite induced increased expression of CCL3 in neutrophils caused higher recruitment of DCs capable of inducing a strong proinflammatory response and elevated co-stimulatory molecule expression compared to LdWT infection. To further illustrate neutrophil-DCs interactions in vivo, we infected LYS-eGFP mice with red fluorescent -/-LdWT/LdCen parasites and sort selected DCs that engulfed the neutrophil containing parasites or DCs that acquired the parasites directly in the ear draining lymph nodes (dLN) 5d post infection. The DCs predominantly acquired the parasites by phagocytosing infected neutrophils. Specifically, DCs containing -/-LdCen parasitized neutrophils exhibited a proinflammatory phenotype, increased expression of costimulatory molecules and initiated higher CD4+T cell priming ex-vivo. Notably, potent DC activation occurred when LdCen-/- parasites were acquired indirectly via engulfment of parasitized neutrophils compared to direct engulfment of LdCen-/- parasites by DCs. Neutrophil depletion in -/-LdCen infected mice significantly abrogated expression of CCL3 resulting in decreased DC recruitment in ear dLN. This event led to poor CD4+Th1 cell priming ex vivo that correlated with attenuated Tbet expression in ear dLN derived CD4+ T cells in vivo.-/-LdCen containing neutrophils phagocytized by DC markedly influence the phenotype and antigen presenting capacity of DCs early on and thus play an immune-regulatory role in shaping vaccine induced host protective response.Collectively, Leishmania is a neglected tropical disease. Leishmania donovani is the principal causative agent of VL in East Africa and the Indian subcontinent whereas in Europe, North Africa, and Latin America VL is mainly caused by Leishmania infantum. No licensed vaccine exists against VL. We have reported previously that live attenuated centrin gene-deleted L. donovani (-/-LdCen) parasite vaccine induced strong innate immunity which leads to a protective Th1 response in animal models. We recently demonstrated that neutrophils play an indispensable role following immunization with LdCen-/- parasites in inducing protective Th1 immune response. However, neutrophils also secrete chemokines that attract other innate cells such as dendritic cells and regulate their activities. In the current study we analyzed the interplay between neutrophils and DCs, and its effects on T cell activation during -/-LdCen infection and compared with wild type parasite (LdWT) infection. We observed that higher recruitment of DCs occurred in -/-LdCen infected mice ear draining lymph nodes compared to LdWT. This recruitment is facilitated by increased secretion of the chemokine CCL3 by neutrophils. A markedly decreased DC recruitment was observed in -/-LdCen infected mice following CCL3 neutralization indicating the key role of neutrophils in DC recruitment. Further, we demonstrated that DCs that ingest -/-LdCen infected neutrophils are better activated than those that acquire the parasites independent of neutrophils. Notably neutrophil depletion in -/-LdCen infected mice also attenuated activation of DCs in the ear dLN that resulted in poor CD4+T cell priming. Our results reveal that interaction between neutrophils and DCs play an important role in shaping proinflammatory immune response induced by a live attenuated Leishmania vaccine.Visceral Leishmaniasis (VL), caused by the protozoan parasites of the genus Leishmania protozoa, is endemic to the tropical and subtropical regions of the world. Over 12 million people are currently infected with Leishmania with an annual incidence of approximately 2 million new cases and ~50, 000 deaths in approximately 100 countries throughout the world was added to the bottom wells for 10 min to release adhered cells from the well and filter. Cells from the lower chambers were then stained with trypan blue solution and counted on a hemocytometer.Supernatant derived from uninfected, or neutrophils cultured under different treatment conditions were placed in the lower compartment of a transwell plate . 106LdWT or -/-LdCen parasites or fluorescent LdWTRFP/ LdCen-/- mCherry by means of a 31-gauge needle in a volume of 10 ml. The uninfected naive/control mice received 1x PBS (diluted from 10x PBS) . To obtain chronically infected mice, C57BL/6 mice were infected 24 wk prior with 105L. donovani metacyclic promastigotes via tail vein. The CD4+ T cells were isolated from these mice using CD4 T cell isolation kit and were used for coculture experiment with DCs at a 1:20 DC/CD4+ T cell ratio.C57BL/6 mice were infected through intradermal route in the ear with 10in vitro from bone marrow progenitors. Briefly, after sacrificing the mice their femurs and tibias were excised, cleaned of tissue, and flushed with RPMI medium . Erythrocytes were depleted by using ACK lysis buffer and isolated bone marrow was cultured with complete RPMI medium supplemented with 10% (v/v) fetal bovine serum (FBS) and 1% penicillin (20 U/ml)/ streptomycin (20 \u03bcg/ml) and 20 ng/mL GM-CSF and IL-4 for 7 days to obtain >75% purity of DCs . In a separate experiment, neutrophils were either remain uninfected or infected with LdWT/-/-LdCen parasites and were treated with LPS (1 \u03bcg/ml) (Sigma Aldrich Cat: L2630) for 8 h. The LPS from the neutrophil culture was rigorously washed out by repeated changes of media and co-cultured with isolated BMDCs for 24h. The conditioned media of BMDC cultures were assayed for mouse cytokine via sandwich ELISA. Culture supernatants were collected at 24h post infection to evaluate cytokine (IL-12) production with the use of sandwich ELISA kit . Costimulatory molecules expression on the surface of DCs were measured by flow cytometry where acquisitions of a million events were performed. The detailed method and antibody clones have been described in detail in the Flow cytometry section of the Materials and Methods.Dendritic cells were cultured Leishmania forward primer 5\u2032- CTATTTTACACCAACCCCCAGT-3\u2032 Leishmania reverse primer 5\u2032-GGGTAGGGGCGTTCTGCGAAA-3\u2032 along with the addition of a fluorescent probe 5\u2032-RAAARKKVRTRCA GAAAYCCCGT-3\u2032 for detection. A Black Hole Quencher moiety attached to the 3\u2032 end and Calfluor Red was coupled to a C6 linker at the 5\u2032 end. Fluorescent Leishmania probe (5\u2032-RAAARKKVRTRCAGAAAYCCCGT-3\u2032) was added in the reaction mixture at a final concentration of 1.5 pmols/\u03bcl. Cycling parameters were like this: preheat at 95\u00b0C for 180 s and then 40 two-step cycles of 95\u00b0C for 10 s and 55\u00b0C for 30 s. To measure the number of Leishmania cells that were represented by a given cycle threshold (Ct) value, a standard curve was made by purification of DNA from na\u00efve mice neutrophils spiked with a known number of parasites.The parasite load was determined in the infected neutrophils as previously described . Briefly-/-LdWT/ LdCen parasites for 6 h at 37\u00b0C in 5% CO2 and washed with the medium, and then the cultures were incubated in RPMI medium for a 16h or 24h post-infection. The culture medium was removed, the slides were air dried and fixed via immersion in absolute methanol for 5 min at room temperature and stained using a Diff-Quick stain set , and intracellular parasite numbers were evaluated microscopically. To measure parasite load in these cultures, a minimum of 300 neutrophils were counted. The results are expressed as percentages of neutrophils that were infected by parasites.For microscopic quantitation of parasite load, neutrophils were infected with the in vivo mAb anti-mouse Ly6G (1A8) (1 mg) or GL113 1 d prior to parasite injection and every 48 h after i.d. injection with the parasites till day 5. At day 5, the efficiency and specificity of the depletions were evaluated on lymph node cell preparation as previously described followed by fixation with a Fixation/Permeabilization Solution Kit for 20 min at room temperature and acquired on an LSR II (BD Biosciences) equipped with 407-, 488-, 532-, and 633- nm laser lines using FACS Diva 6.1.2 software. Acquisitions of a million events were performed. Data were analyzed with FlowJo software version 9.7.5 (Tree Star). For analysis, first doublets were removed using width parameter; dead cells were excluded based on staining with the Live/Dead Aqua dye. Lymphocytes were identified according to their light-scattering properties. CD4 T cells were identified as CD3Statistical analysis of differences between means of groups was determined either by unpaired two-tailed Student t test or one- way ANOVA (with a post-test correction for type II error) or two-way ANOVA, using GraphPad Prism 5.0 software. A p value < 0.05 was considered significant, and a p value <0.005 was considered highly significant.L. major infection in mice [-/-LdCen infection in vitro and compared with LdWT infection. Infection of neutrophils with either LdWT or -/-LdCen parasites resulted in an increase in CCL3 mRNA level by an average of 3-fold and 7-fold over uninfected neutrophils respectively of MHCII in ear dLN of -/-LdCen infected mice whereas no significant reduction was observed in LdWT infected mice after CCL3 depletion as shown by the gating strategy and individual flow plots (+Cd11c+MHCIIhiCD8a-CD205low) in ear dLN of -/-LdCen infected mice whereas no significant reduction was observed in LdWT infected mice after CCL3 depletion (We next confirmed the specificity of neutrophil derived CCL3 in the recruitment of DCs to ear dLN via depleting CCL3 ondition . Interesow plots and subsow plots . Furtherepletion .-/-LdCen infected mice and compared with LdWT infected mice. Parasitized DCs were sort selected from different groups of mice ear dLNs by gating live single cells (Cd11b+Cd11c+ MHCIIhi RFP/mCherry+) (-/-LdCen group ear dLN showed a significantly higher level of CD80 and CD40 expression (LdWT group (It is possible that DCs may arrive at the site of infection by homeostatic migration in addition to active recruitment by chemokine gradients. We therefore characterized the phenotype of DCs recruited/migrated to the ear dLN in Cherry+) and assepression and prodpression with an WT group .in vivo, fluorescent LdWTRFP or LdCen-/- mCherry parasites were injected into the ear dermis of LYS-eGFP mice which enables identification of neutrophils by GFP expression. We observed two types of DCs from the ear dLN after sorting; viz; Cd11b+Cd11c+MHCIIhieGFPhiRFP+ and Cd11b+Cd11c+MHCIIhieGFP-RFP+ . The sorting strategies are shown in LdWT and -/-LdCen infected mice ear dLNs thereby suggesting that the majority of the DCs acquired the parasites via uptake of infected neutrophils as shown in the schematic diagram in nfection . Interested mice . Next, wted mice . These rLdWT or -/-LdCen infected mice following GL113 or 1A8 treatment. The sorting strategy is identical as shown in +Cd11c+Ly6G/Ly6C-MHCIIhi) were sort selected from PBS injected mice. Neutrophil depletion significantly decreased the proinflammatory cytokine gene expression such as IL-12 were measured. Corroborating our previous observation in + cells in the ear dLNs of -/-LdWT/LdCen infected mice and thus fail to suppress DC activation. Indeed, we observed that -/-LdCen infected neutrophils induced enhanced DC activation as indicated by elevated expression of costimulatory molecules concomitant with an induction of pro-inflammatory IL-12 secretion which may result in the increased capacity of these DCs to induce Th1 cell proliferation.The key role of neutrophils in DC recruitment through the synthesis of chemokines has been documented in several parasitic and bacterial infections . For exand CCL20 . Likewisthe dLNs . Notablyion site . In our esponses . We obsein vitro results described above were further validated by in vivo experiments in an intradermal ear model of infection. Consistent with our in vitro results, we also observed that neutrophils responding to \u2212/\u2212LdCen parasites in ear dLN augmented CCL3 expression, which played a critical role in attracting DCs 5d post infection. Interestingly, in -/-LdCen infected mice neutrophil derived CCL3 also has a predominant role in the migration of Cd11b+MHCIIhiCD11c+CD8\u03b1\u2212CD205lowDC, the major DC population that primes CD4+ T cell response in dLNs previously described in L major studies [-/-LdCen immunization has been shown to be mediated by both CD4+ and CD8+ T cells [+CD11c+Ly6G/Ly6c- MHCIIhiDC population in the vaccine context. Although, same number of total myeloid /Cd11b+ cells migrated to the ear dLN in both groups of infected mice, neutrophils from -/-LdCen infected mice were potent in achieving a heightened DC migration in dLN due to elevated CCL3 production. It is worth mentioning here, besides these subtypes of DCs studied by us, recent study highlights monocyte-derived dendritic cell-like phagocyte subset (Ly6C+CCR2+ monocytes with high CD11c expression) serve as a predominant reservoir for efficient intracellular multiplication of L. major and spread to the new host cells [Leishmania vaccine immunity warrants further investigation.The studies . However T cells implyingst cells . However\u2212/\u2212LdCen parasites in the ear (the site of parasite inoculation) augmented the expression of CCL3 which attracted significantly higher number of dermal DCs 24 h post infection. However, DCs are professional APCs playing a key role in launching and regulation of the immune response via interacting with T cells that occurs in lymph nodes (LNs) [LdCen-/- specific immune response and thereby, on the out-come of infection.Notably, we also observed that neutrophils responding to es (LNs) . Hence, L. major infection acquire their infections via capture of infected neutrophils, and the sequestration of L. major antigens within apoptotic neutrophils would seem an especially efficient process for inhibiting DC activation and their ability to initiate the Leishmania-specific T-cell response [+ T cell activation [M. tuberculosis by delivering M. tuberculosis to DCs in a form that makes DCs more effective initiators of naive CD4 T cell activation [LdWT or -/-LdCen infected neutrophils by DCs in the ear dLN 5d post infection of LYS-eGFP mice. The majority of the Cd11c+DCs (termed P1 DCs) recovered from the ear dLN at 5d post infection were positive for both the RFP and eGFP signals and stained positive for neutrophil markers. This demonstrated that the majority of the infected DCs acquired their parasites via engulfment of infected neutrophils similar to studies in M. tuberculosis [-/-LdWT/LdCen parasite infected neutrophils were better activated than the DCs (termed P2 DCs) that acquire the parasites independent of neutrophils. Interestingly, DCs that ingested -/-LdCen infected neutrophils in ear dLN 5d post infection were higher in numbers and showed significant induction of costimulatory molecules and proinflammatory cytokines compared to DCs bearing LdWT infected neutrophils. We further addressed the consequences of -/-LdWT/ LdCen infected neutrophil capture by DCs on the latter\u2019s ability to present parasite-derived antigen to CD4+ T cells ex-vivo. Corroborating with our in vitro observation, unlike LdWT parasites, -/-LdCen parasite infected neutrophils were defective in their capacity to attenuate the activation of DC. Thus, significant enhancement of DC mediated CD4+T cell activation following capture of -/-LdCen infected neutrophil occurred compared to DCs that ingested LdWT infected neutrophils. In either case it was significantly higher than respective DCs that acquire the parasites directly. Consequently, our data draw a clear distinction between the ability of -/-LdCen infected and LdWT infected neutrophils to deliver activation signals to DCs in the ear dLNDuring pathogenic infection, neutrophils and DCs, normally positioned in distinct anatomical compartments, may colocalize at sites of inflammation. Neutrophils are short-lived cells that must be targeted for orderly removal upon apoptotic death. The function of DCs in the clearance of dying neutrophils is central to the resolution of infection that will in most instances affect antigen-presenting properties of DCs . Generalresponse . In conttivation ,41. For tivation . In our rculosis . Notably-/-LdCen containing neutrophils was essential for -/-LdCen to enhance DC maturation and to initiate the anti-Leishmania CD4+T cell response. The less potent immune response observed in P2 DCs that directly acquired either LdWT or LdCen-/- parasites highlights the novel findings from our study. Further, our data shows that neutrophils may not simply be an efficient mechanism of parasite delivery as illustrated in other pathogens but that their immunoregulatory role in enhancing the DCs activation towards APC activity is critical in producing potent anti-parasitic mechanisms induced by LdCen-/- infection.Further, the uptake of -/-LdCen parasite mediated immunity was further validated through the observation that depletion of neutrophils results in decreased recruitment of DCs to dLNs. This finding is in accordance with the study by Charmoy et al. 2010 indicating that neutrophils are essential for DC recruitment to the ear dermis following L. major inoculation [-/-LdCen infected mice as indicated by the presence of lower percentages of activated DCs (CD80+ and CD40+) along with attenuated expression of cytokines such as IL-6, TNF and heightened expression of IL-10 which subsequently compromised the proliferation of naive Ag-specific CD4+ Th1 cells ex-vivo. The consequence of this inhibition in Leishmania specific host protective CD4+ Th1 cell priming was directly supported by the attenuated Tbet expression in ear dLN derived CD4+ T cells in neutrophil depleted mice consistent with the studies in M. tuberculosis infection where depletion of neutrophils resulted in decreased migration of DCs to dLNs and delayed activation and proliferation of naive Ag-specific CD4+ T cells in dLNs of mice [-/-LdCen infected mice. However, it has been reported earlier that neutrophils expressing CCR7 can migrate to the lymph nodes via the lymphatics rendering activation/maturation of DCs via upregulation of MHC-II, CD40, CD80 and CD86 molecules as well as IL-12, TNF-\u03b1 and IL-6 secretion [-/-LdCen infected neutrophils compared to LdWT infected neutrophils [-/-LdCen infection significantly higher number of neutrophils migrated to the lymph nodes and resulted in higher DC activation/maturation compared to LdWT as indicated by significant upregulation of maturation marker/costimulatory molecules which was abrogated in neutrophil depleted mice. Further, significantly higher DC numbers in ear dLN of -/-LdCen infected mice indicate that neutrophil derived CCL3 generation facilitated increased DC migration and subsequent recruitment to the dLN of these mice. Notwithstanding, we will be pursuing this detailed line of research in the future. Collectively, neutrophils in -/-LdCen infected mice facilitate enhanced DC migration and subsequent recruitment along with DC activation/maturation in dLN and confers them with the capacity for efficient priming of Th1 cells.To summarize, the importance of neutrophils-DC interaction in culation . It is iculation , depletiecretion . Previou+ Th1 cells involved in protection by live attenuated Leishmania vaccine against virulent L. donovani challenge has been reported recently by our group [+ T cell immune response by affecting the function of DCs during -/-LdCen infection and such cross talk involving neutrophils and DCs results in significantly lower -/-LdCen parasite numbers at early time point via enhanced effector response and might cumulatively contribute towards generating live attenuated Leishmania vaccine\u2013induced protective immunity as observed earlier [LdWT parasites utilizes such neutrophil-DC crosstalk to subvert the host effector response which enables parasite proliferation in the host.The role of neutrophils in providing direct stimulatory cues to effector CD4ur group . Additio earlier . ConversLdWT and -/-LdCen parasites deploy differential immune strategies to modulate DC function. While LdWT parasite infected neutrophils cause diminution of DC activation, -/-LdCen parasite infected neutrophils interact with DC, generates activation of adaptive immune response. Thus, the impact of the early neutrophil- DC interactions described in this study clearly highlights the immunological differences exist between a virulent and live attenuated parasite in modulation of DC response and may be especially relevant in future vaccine development against Leishmaniasis.In summary, our results indicate that S1 FigA) qRT-PCR assay showing the parasite burden in the neutrophils infected with LdWT or \u2212/\u2212LdCen parasites for 16h. (B) Peritoneal neutrophils were infected with -/-LdWT/LdCen parasites for 16h. Intracellular parasite numbers were visualized by Giemsa staining and estimated microscopically. The infection efficiency (percentage of infected cells) was recorded. (C) qRT-PCR assay showing the parasite burden in the neutrophils infected with LdWT or \u2212/\u2212LdCen parasites for 24h. (D) Peritoneal neutrophils were infected with -/-LdWT/LdCen parasites for 24h. Intracellular parasite numbers were visualized by Giemsa staining and estimated microscopically. The infection efficiency (percentage of infected cells) was recorded. To measure parasite load in these cultures, a minimum of 300 neutrophils were counted. The data represent the mean values \u00b1 SD of results from three independent experiments that all yielded similar results. Peritoneal neutrophils were either left uninfected or infected with -/-LdWT/LdCen parasites for 16h. Changes in the mRNA expression levels of CCL4 and CCL5 from uninfected and infected neutrophils were determined by qPCR as described in Materials and Methods. The data represent the mean values \u00b1 SD of results from three independent experiments that all yielded similar results. *p< 0.05; ** p < 0.005. (G) BMDCs were cocultured with LPS activated uninfected or infected neutrophils as described in Materials and Methods. The percentages of MHCII and CD40 positive DCs were reported. The data represent the mean values \u00b1 SD of results from three independent experiments that all yielded similar results. *p< 0.05; ** p < 0.005 *** p < 0.0005.((PDF)Click here for additional data file.S2 FigLdWT or LdCen-/-. (A)No significant differences in the CCL4 and CCL5 mRNA expression was observed in sort selected parasitized neutrophils from ear dLN 5d post infection with The FMO\u2019s for all the fluorophores used in experiment (B&J) have been shown and are based on the same gating as in (B&J). (B) Sorting strategy showing parasitized neutrophils isolation from ear dLN. (C) Post sort analysis for the sort selected parasitized neutrophils has been shown. (D) Confocal microscopic image of the sort selected parasitized neutrophils stained with Hoechst nuclear dye with excitation 561 and 405 nm for red and blue channel have been shown. Scale bar 1\u03bcm. (E) Confocal microscopic image of the sort selected parasitized neutrophils with excitation 670, 590 and 405 nm for alexa700, mRFP and Pacific blue channels imaging respectively have been shown. Scale bar 2\u03bcm. mRNA expression levels of CCL4 and CCL5 from sorted neutrophils in the ear dLN were estimated 5d post infection by qPCR and expressed as fold change from uninfected naive mice (n = 5). The experiment was repeated three times with pooled digests from eight to twelve ear dLNs per experiment. The data represent the mean values \u00b1 SD of results from three independent experiments that all yielded similar results. (H) The gating strategy and (I) changes in the total number of DCs (CD11b+CD11c+MHCIIhiCD205lowCD8\u03b1-) per ear dLN have been shown under CCL3 non-depleted or depleted condition 5d post infection. Values shown are the mean numbers of cells per ear dLN \u00b1 SD of results. 6\u20138 ear dLN, pooled data from 3 independent experiments that all yielded similar results (n = 6). *p< 0.05; ** p < 0.005 *** p < 0.0005. (J) Parasitized DCs were flow sorted from the ear dLN 5d post infection. The sorting strategy has been displayed. (K) Post sort analysis for the sort selected parasitized DCs has been shown. (L) Confocal microscopic image of the sort selected parasitized DCs have been shown. Scale bar 2\u03bcm.(PDF)Click here for additional data file.S3 Fig(A) Gating strategy for flow cytometry analysis and representative flow plots showing the expression of CD40 in P1 and P2 DCs in ear dLN of LYS-eGFP infected mice. (B) Ag-specific CD4 T cell proliferation was estimated from P2 DC\u2013CD4 T cell coculture assay by studying CFSE dilution of gated CD4+CD44+ T cells and is represented by the histogram.(PDF)Click here for additional data file.S4 Fig(A) The gating strategy and individual flow plots for DCs (Cd11b+Cd11c+Ly6G-Ly6C-MHCIIhi) recruitment in ear dLN has shown at 5-day post infection in mice depleted or not of neutrophils.(PDF)Click here for additional data file.S5 Fig(A) Total number of Cd11b+ cells recruited in ear dLN 5 days post infection in either GL113 or 1A8 treated mice has been shown. The experiment was repeated three times with pooled digests from five to six ear dLNs per experiment. The data represent the mean values \u00b1 SD of results from three independent experiments that all yielded similar results. The percentages of CD80 and CD40 positive DCs in ear dLN of GL113/1A8 treated -/-LdWT/LdCen infected mice 5d post infection was reported. (D) Gating strategy showing CFSE dilution of gated CD4+CD44+T cells from DC-CD4 T cell coculture assay.(PDF)Click here for additional data file.S6 Fig(A) Schematic diagram showing experimental scheme for neutrophil depletion and subsequent experiments. (B) Real-time PCR analysis of RNA isolated from purified CD4 LN T lymphocytes at 5d post infection as described in Materials and Methods is shown. Normalized expression levels of Tbet were estimated. Data are presented as fold change from uninfected naive mice. The data represent the mean values \u00b1 standard deviations of results from 3 independent experiments that all yielded similar results. *, P < 0.05; **, P < 0.005. (n = 6). (C) Parasite numbers in ear dLNs of different groups of GL113/1A8 treated infected mice were measured 7 days post infection. Means and standard errors of the means for 5 mice in each group are shown. Data are representative of two independent experiments. *, P < 0.05.(PDF)Click here for additional data file."} +{"text": "ABSTRACT IMPACT: Our work might lead to a new treatment for patients with acute myeloid leukemia OBJECTIVES/GOALS: Acute myeloid leukemia (AML) is a devastating hematologic malignancy, with dismal 5-year survival. Chimeric antigen receptor (CAR) T cells have been approved for B cell malignancies but not for AML. The goal of this study is to explore the safety and efficacy of CAR T cells targeting CD105 (endoglin) to treat AML. METHODS/STUDY POPULATION: We have constructed human and murine CAR T cells targeting CD105. The CARs were created by sequencing the V(D)J regions of hybridomas and designing single chain variable fragments that target CD105 which were subsequently introduced in a CAR backbone via Gibson assembly. The CAR T cells were produced via transduction using retrovirus or lentivirus. Leukemia cell lines were assessed for CD105 expression with flow cytometry. Killing assays were performed via measurement of luminescence of target cells after co-culture with CAR T cells. Activation assays were performed with co-culture of CAR T cells and target cells and measurement of activation markers with flow cytometry. To assess in vivo efficacy and safety, murine CAR T cells were infused into C57BL/6J mice carrying B16 melanoma after lymphodepletion. RESULTS/ANTICIPATED RESULTS: All human leukemia cell lines assessed expressed some degree of endoglin apart from the T cell leukemia Jurkat. Human CD105 CAR T cells were activated by co-culture with leukemia cell lines and effectively killed leukemia cells in vitro in a CD105-specific manner. Murine CAR T cells killed efficiently both murine solid tumors (B16 melanoma) and murine leukemias (C1498) in vitro. Murine CAR T cells did not exhibit any toxicity when infused after low-dose lymphodepletion (cyclophosphamide 100mg/kg) but caused significant morbidity after higher doses (cyclophosphamide 200mg/kg). Murine CAR T cells delayed the growth of B16 melanoma in immunocompetent mice. DISCUSSION/SIGNIFICANCE OF FINDINGS: We have constructed human and murine CD105 CAR T cells with excellent activity in vitro. The activity of human CD105 CAR T cells in xenografts and the biologic relevance of the toxicity of murine CD105 CAR T cells in humans needs to be further investigated. CD105 CAR T cells might prove an important therapeutic option for patients with AML."} +{"text": "What are the comparative costs and benefits of physician continuous professional development (CPD) for drug prescribing?In this systematic review of 38 studies, CPD was associated with reduced health care costs (median drug cost savings of $79\u2009373) compared with no training. More intensive CPD (compared with no training or less intensive interventions) was associated with improved effectiveness (prescribing) outcomes but incurred greater education costs .These results suggest that physician CPD for drug prescribing is associated with reduced health care costs and that both effectiveness outcomes and costs should be considered when making education decisions. The economic impact of continuous professional development (CPD) education is incompletely understood.To systematically identify and synthesize published research examining the costs associated with physician CPD for drug prescribing.MEDLINE, Embase, PsycInfo, and the Cochrane Database were searched from inception to April 23, 2020, for comparative studies that evaluated the cost of CPD focused on drug prescribing. Two reviewers independently screened all articles for inclusion and reviewed all included articles to extract data on participants, educational interventions, study designs, and outcomes (costs and effectiveness). Results were synthesized for educational costs, health care costs, and cost-effectiveness.Of 3338 articles screened, 38 were included in this analysis. These studies included at least 15\u2009659 health care professionals and 1\u2009963\u2009197 patients. Twelve studies reported on educational costs, ranging from $281 to $183\u2009554 . When economic outcomes were evaluated, 31 of 33 studies (94%) comparing CPD with no intervention found that CPD was associated with reduced health care costs (drug costs), ranging from $4731 to $6\u2009912\u2009000 . Four studies found reduced drug costs for 1-on-1 outreach compared with other CPD approaches. Regarding cost-effectiveness, among 5 studies that compared CPD with no intervention, the incremental cost-effectiveness ratio for a 10% improvement in prescribing ranged from $15\u2009390 to $437\u2009027 to train all program participants. Four comparisons of alternative CPD approaches found that 1-on-1 educational outreach was more effective but more expensive than group education or mailed materials .In this systematic review, CPD for drug prescribing was associated with reduced health care (drug) costs. The educational costs and cost-effectiveness of CPD varied widely. Several CPD instructional approaches were more effective but more costly than comparators. This systematic review of 38 studies assesses the association between the costs of physician continuous professional development and drug costs related to prescribing. Systematic reviews have found that restrictive interventions to optimize prescribing have benefits8 but may be associated with treatment delays and adverse effects on clinicians\u2019 professional identity and culture.6 By contrast, educational interventions have overall favorable effects on practitioner performance and patient outcomes.14Inappropriate prescribing harms patients and wastes resources.14 of interventions to improve drug prescribing have touched only briefly on the economic outcomes of CPD interventions. Only 1 review15 (published in 2002) focused on cost of CPD, and that review included only 1 study of CPD for drug prescribing.Continuous professional development (CPD) is essential to clinicians\u2019 efforts to maintain competency after completion of training and includes formal educational interventions and unstructured learning activities. Although CPD programs to improve prescribing practices are increasingly common, their cost and cost-effectiveness remain incompletely characterized. ReviewsA comprehensive synthesis of evidence regarding the comparative costs and benefits of physician CPD for drug prescribing would provide clinicians, educators, and administrators information to reduce wasted effort (cost and physician time), identify resource-efficient instructional approaches, and promote more effective health care. We conducted a systematic review to determine the comparative costs and benefits of physician CPD for drug prescribing and the CPD features that are associated with improved cost-benefit outcomes.16) was planned, conducted, and reported in adherence to the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) reporting guideline17 and reporting guidelines for economic evaluations.20This systematic review (part of a larger systematic review of economic outcomes of CPDphysicians), intervention , and outcomes21 . We used this strategy credit is awarded; or self-directed learning or self-assessment activities for which CME credit is awarded.\u201dReviewers worked in assigned pairs to screen studies for inclusion, first reviewing the title and abstract and then reviewing the full text if needed . Reviewers resolved all disagreements by consensus.22 organizing costs according to defined categories16 related to education planning and implementation. We resolved conflicts by consensus.We implemented a data abstraction form in software designed for systematic reviews . Two reviewers (D.A.C. and C.R.S.) independently extracted information on study design, participants, CPD interventions, methodological quality, effectiveness outcomes , educational costs, and health care costs. For educational costs, we used Levin\u2019s \u201cingredients\u201d approach,23 which evaluates the study design, sampling, outcomes , and statistical analyses. We also appraised methods specific to economic analyses, selected from the 1996 BMJ guidelines.24We appraised general methodological quality using the Medical Education Research Study Quality Instrument,28), we did not attempt such analyses. Instead, to evaluate cost-effectiveness, we adopted the dominance ranking matrix approach29 advocated by the Joanna Briggs Institute.27 We also calculated monetary net benefit for all studies that reported educational costs and health care costs.We converted all monetary estimates to 2021 US dollars, first adjusting for inflation in the original currency and then converting the original currency into US dollars using exchange rates on May 6, 2021. Given well-documented concerns about meta-analysis of cost-effectiveness studies and collected data on more than 1\u2009963\u2009197 patients . Three studies62 were translated from a non-English language (2 German and 1 Spanish). Five studies included more than 2 arms,59 resulting in more than 1 relevant comparison per study. After describing overall study features and quality, we report educational costs, health care cost impact, and educational costs combined with effectiveness outcomes (cost-effectiveness and net benefit).We identified 3338 potentially eligible studies of which 38 met criteria for inclusion and studies that evaluated health care costs after a CPD activity (35 studies67).66 (34%) involved other health care professionals (besides physicians), including postgraduate physician trainees (n\u2009=\u20095), nurse practitioners and physician assistants (n\u2009=\u20095), pharmacists (n\u2009=\u20095), nurses (n\u2009=\u20093), and medical students (n\u2009=\u20091).All studies involved physicians in independent practice, most of whom were family, internal, or general medicine physicians (26 studies [68%]) or all physicians in the facility (4 studies [11%]). Thirteen studies66 compared 2 approaches to increase attendance at a CPD course on drug prescribing.Most studies focused on optimal use of specific drugs or drug classes, including antibiotics (n\u2009=\u200915 studies), analgesics (n\u2009=\u20093 studies), antihyperlipidemics (n\u2009=\u20093 studies), and neuropsychiatric drugs (n\u2009=\u20093 studies). Three studies focused on general drug-drug interactions or prescribing errors. One study67), face-to-face small groups or 1-on-1 outreach (22 studies65), performance audit and feedback (19 studies65), and face-to-face large groups (14 studies65). Four studies67 also implemented changes in clinical practice .Instructional modalities included paper materials (26 studies67 (32 [84%]); 6 (16%) were in academic settings57 and 2 (5%) in government facilities.62 Thirty studies67 (79%) involved patients in an ambulatory setting, 7 (18%) involved patients in a hospital ward,62 and 3 (8%) involved a medical procedure.39 Geographic origins included the US (15 [39%]), the UK (5 [13%]), other European countries (11 [29%]), Asia (3 [8%]), Canada (3 [8%]), and Oceania (1 [3%]). Funding sources included industry61 (5 [13%]), government66 (10 [26%]), private53 (2 [5%]), and local57 (1 [3%]) support. Twenty studies (53%) did not report funding67; among these, for 5 studies44 the authors were employed by a capitated health plan .Considering the context of education, most studies were conducted in private or independent practice65 12 (32%) used 2 or more nonrandomized groups,67 and 14 (36%) involved a single-group pre/post or time-series design.62 Nearly all studies (35 [92%]) compared CPD against no intervention67 (baseline performance or a no intervention control group), whereas 10 (26%) compared one CPD intervention against another66 .Twelve studies (32%) used randomized group assignment,67 (58%). Evidence to support the validity of the outcome measure was infrequently reported: 10 studies (26%) reported content evidence67 and 5 (13%) reported internal structure evidence64 (such as interrater reliability); none reported correlation with another variable. Participant retention was high in 27 studies67 (71%) . Thirty-two studies67 (84%) specified the time horizon, 3 (8%) stated a discount rate24 or justified its absence,63 and 3 (8%) conducted a sensitivity analysis.64 See eTable 2 in the Outcome measures were blinded or obtained from an unbiased data source in 22 studies66 (32%) reported costs associated with implementing CPD interventions. For each study, we extracted 12 possible cost ingredients22 (63 (25%) reporting details of ingredient quantification and 5 studies63 (42%) reporting details of pricing.Twelve studiesdients22 . The ing45 that analyzed only postage and paper and reached 36 physicians) to $183\u2009554 , with a mean (SD) of $32\u2009676 ($51 215) .The mean ingredient cost per intervention varied widely, ranging from $58\u2009897 for learner attendance to $1022 for catering (mean of 2 interventions). The total reported educational cost likewise varied widely, ranging from $281 (a study67 (92%) evaluated health care costs after a CPD activity , 10 studies67 encouraged choice of less expensive drugs, 7 studies58 targeted fewer prescriptions overall, and 7 studies64 encouraged increased use of an effective treatment.Thirty-five studies67 reported an outcome of drug cost, and this outcome was often the only health care cost outcome; thus, we used drug cost as the (common) metric of health care costs. For 1 study56 the investigators expected higher short-term drug costs if prescribing recommendations were followed (ramipril in patients with diabetes); for the other 34 studies,67 the economic goal appeared to be immediate cost savings. The appropriateness of drug prescription was appraised in 5 studies67 (14%). Twenty-five studies65 took steps to ensure an appropriate (less biased) comparison by using a concurrent control group65 (10 [29%]) or making historical comparison matched for time of year65 (15 [43%]). Twenty-four studies67 (69%) reported details of resource quantification, and 26 studies67 (74%) reported details of pricing. Thirty studies67 (86%) reported the period of data collection, which ranged from 1 to 182 weeks.All 35 studies67 [n\u2009=\u200923 comparisons] or baseline metrics62 [n\u2009=\u200914 comparisons]). In all but 2 instances,65 these studies found that CPD was associated with lower drug costs (56 CPD led to increased prescribing of ramipril as intended for patients with diabetes and a concomitant (expected) increase in drug costs. In the other study,65 the rate and cost of antibiotic prescriptions were reduced as intended, but recommendations for nonantibiotic symptomatic treatments increased, resulting in higher total drug costs.Thirty-three studies (37 comparisons) evaluated drug costs for CPD compared with no intervention , and the percentage of savings compared with the control group or baseline costs ranged from 2.8% to 62.0% (n\u2009=\u200929 studies). Substantial variation in study time period and sample size precluded meaningful cross-study comparisons of total costs, so when possible we standardized these variables to reflect a fixed period and number of patients or physicians (100 for 1 year). Although admittedly imperfect , this approach offers useful insights. Savings for 100 patients ranged from $1.00 to $186\u2009862 per year (n\u2009=\u200924 studies), and savings for 100 physicians ranged $11\u2009058 to $28\u2009800\u2009000 per year (n\u2009=\u20095 studies).Among the remaining 31 studies,65 (9 comparisons) compared drug costs for 2 or more alternative CPD approaches 65 and year-round activities.55 Contrary to their authors\u2019 hypotheses, studies found higher drug costs for group outreach 54 and for 1-on-1 outreach augmented with vivid case studies .33Seven studiesproaches . Four st63 (24%) reported costs and clinical (prescribing) outcomes for CPD compared with no education (a no intervention control group or baseline metrics). Eight of these studies63 reported an effectiveness outcome , thus allowing estimation of the incremental cost-effectiveness ratio (ICER). All these studies63 found improved prescribing outcomes with CPD, but costs were of course higher than for no education . When further standardized to 1 physician trained, the ICERs ranged from $152 to $3441 per physician. Standardized ICERs for other outcomes ranged from $179 to $3258 per physician.63 (11%) reported information that allowed estimation of the net benefit for CPD compared with no education. Three studies38 found a favorable net benefit , but extrapolation of benefits suggested a break-even point approximately 3.5 years after the intervention.Four studies benefit . The fou60 (13%) (6 comparisons) compared the costs and clinical (prescribing) outcomes of 2 or more alternative CPD approaches found that 1-on-1 educational outreach was more effective but more expensive than group education or mailed materials (ICERs ranging from $18 to $4105 per physician trained). Two other studies60 compared face-to-face instruction with mailed materials; one study60 found that face-to-face instruction was more effective but more expensive (ICER of $3 per physician trained), and the other study31 found a net benefit of $221 per physician favoring face-to-face instruction. One additional study66 compared 2 approaches to recruit participants to a CPD activity on drug prescribing and found that informal publicity through local organizations was more effective (higher attendance) and less expensive than a formal advertising campaign .Studies that compare the cost-effectiveness of alternative educational approaches are vital to efforts to optimize CPD. Five studiesproaches . All the67 that evaluated health care costs (drug costs), we found that CPD is associated with substantial cost savings. Among studies66 that evaluated educational costs, cost-effectiveness estimates varied widely, even after attempts to standardize the outcome measures. Along with expected differences in local contexts, educational approaches, and effectiveness outcomes, this heterogeneity arises from large differences in the accounting of educational costs. For example, only 4 studies accounted for 5 cost ingredients or more in their cost estimates, and 3 studies reported no discrete ingredients. Across all studies, we found that different CPD approaches can be associated with substantial differences in health care costs, educational costs, and incremental cost-effectiveness.This systematic review examined the costs and economic benefits of CPD for practicing physicians. Among studiesStudies reported information on participants (physicians and patients) inconsistently. Studies likewise frequently omitted details on ingredient (resource) quantification and pricing for educational costs (missing in >50%) and drug costs (missing in >25% of studies).14 of drug prescribing have suggested that outreach, audit and feedback, and multimodal educational interventions are consistently associated with improved drug prescribing . Our review extends these findings by examining cost along with effectiveness and confirms that 1-on-1 educational outreach is effective but incurs higher cost. Audit and feedback was frequently used in the studies we identified, but study designs did not allow direct cost comparisons.Previous reviews67; median immediate savings of $79\u2009373). Two studies65 found increased costs, but in 1 study56 this was intentional , and in the other65 there was a decrease in the prescription rate and cost of the target drug (antibiotics) despite an increase in overall drug costs. These exceptions underscore the importance of explicitly stating and justifying the intended impact and illustrate that focused, immediate outcomes may differ from broad, delayed economic measures.The findings of this review support 4 important messages. First, CPD was associated with substantially reduced drug costs than others. We observed a 50-fold difference in highest vs lowest mean cost per ingredient. The most expensive ingredients reflected time (opportunity cost of lost wages or productivity) invested by learners, administrators, and faculty, and facility costs. Indeed, itemized time expenses collectively represented 55% of all educational costs , and the72 Our method to standardize educational costs, cost-effectiveness, and net benefits to a common time period, sample size, and level of effectiveness offers an example that others may choose to adopt when synthesizing economic data. However, this synthesis would have been facilitated by more complete accounting of educational costs,22 more consistent measurement of clinical outcomes , and better reporting71 in the original studies. We encourage education researchers to measure and report costs more frequently, robustly, and completely.Fourth and most importantly, our findings amplify prior requests for more and better educational cost evaluations.27 and a variety of tabular summaries.This study has several strengths, including the robust literature search, duplicate review at all stages, and synthesis of key study findings using recommended approaches68 Moreover, differences in local systems, funding, culture, and values will constrain local applications of cost-related outcomes regardless of rigor in the original data collection or meta-synthesis.68 However, we propose that syntheses such as our can help establish boundaries of plausible outcomes that decision makers can apply locally. Finally, our findings regarding what works to enhance CPD effectiveness were limited by the paucity of studies and the wide variation in research questions.This review also has some limitations. This study was limited by poor reporting in many original studies, which impaired our ability to identify key elements of methods and results. In particular, data on the number of physicians trained or patients treated were reported inconsistently, such that it was not always possible to rescale data to a common unit that allowed straightforward synthesis or comparison across studies. In addition, both effectiveness and costs are sensitive to numerous design features, including sample size, outcome selection and measurement, and duration of follow-up . Robust standardization would require a direct association among cost, effectiveness, and scale , which is unlikely.In this systematic review, CPD for drug prescribing was associated with reduced health care (drug) costs. The educational costs and cost-effectiveness of CPD varied widely. Several CPD instructional approaches were more effective but more costly than comparators."} +{"text": "To determine the prevalence of emotionally unstable personality disorder (EUPD) attending a community mental health team (CMHT) in a major Irish cityTo describe the current psychiatric care afforded to this cohort of service userClinical chart review of all 328 patients attending a CMHT outpatient in an urban setting was carried out. Patients diagnosed with EUPD or displayed features of EUPD were identified. Data on the various interventions offered to this cohort of service users were collected and compared against current guidelines.Out of the 328 patients actively attending the service, almost 17% (n = 55) were diagnosed with EUPD and further 6% (n = 19) were found to display features of EUPD such as emotional dysregulation, self-harming behaviour and cognitive distortions. Comorbid psychiatric disorder such as mood or anxiety spectrum disorder was diagnosed in 23% (n = 17) of this cohort. Meanwhile, 8% (n = 6) was diagnosed with addiction disorders and 5% (n = 4) was diagnosed with a comorbid personality disorder. A significant proportion of 77% (n = 57) were prescribed psychotropic medication with 51% (n = 29) being on more than one psychotropic medication. Antidepressants, antipsychotics and hypnotics were the three most common medications prescribed at the rate of 89% (n = 51), 30% (n = 17) and 28% (n = 16) respectively. A majority of 66% (n = 49) were offered intervention from a multi-disciplinary team (MDT) member with 47% (n = 23) being offered more than one type of intervention. Referrals to community mental health nurses and psychology service were the two most common interventions offered with a referral rate of 59% (n = 29) and 55% (n = 27) respectively. 28% (n = 21) of service users with EUPD or EUPD traits has had at least one hospital admission while attending the CMHT and 46% (n = 34) have been admitted to the day hospital at least once.The prevalence of EUPD in our outpatient sample corresponds with findings in previous studies. Standard psychiatric care is the most common option available to the majority of general adult patients with EUPD in Ireland due to the lack of any national treatment programme and scarce availability of specialised therapeutic approaches such as dialectical behavioural therapy within community mental health teams. Our CMHT will attempt to integrate mentalization-based treatment into our outpatient management of EUPD patients taking into account current clinical guidelines for management of EUPD and resources needed for training and delivering the intervention."} +{"text": "Traditional approaches for SSI surveillance have deficiencies that can delay detection of SSI outbreaks and other clinically important increases in SSI rates. Optimized SPC methods for SSI surveillance have not been prospectively evaluated.We conducted a prospective multicenter stepped wedge cluster RCT to evaluate the performance of SSI surveillance and feedback performed with optimized SPC plus traditional surveillance methods compared to traditional surveillance alone. We divided 13 common surgical procedures into 6 clusters (Table 1). A cluster of procedures at a single hospital was the unit of randomization and analysis, and 105 total clusters across 29 community hospitals were randomized to 12 groups of 8-10 clusters . After a 12-month baseline observation period (3/2016-2/2017), the SPC surveillance intervention was serially implemented according to stepped wedge assignment over a 36-month intervention period (3/2017-2/2020) until all 12 groups of clusters had received the intervention. The primary outcome was the overall SSI prevalence rate (PR=SSIs/100 procedures), evaluated with a GEE model with Poisson distribution.Table 1Figure 1Schematic for stepped wedge design. The 12-month baseline observation period was followed by the 36-month intervention period, comprised of 12 3-month steps.Our trial involved prospective surveillance of 237,704 procedures that resulted in 1,952 SSIs (PR=0.82). The overall SSI PR did not differ significantly between clusters of procedures assigned to SPC surveillance and those assigned to traditional surveillance (Table 2). SPC surveillance identified 104 SSI rate increases that required formal investigations, compared to only 25 investigations generated by traditional surveillance. Among 10 best practices for SSI prevention, 453 of 502 (90%) SSIs analyzed due to SPC detection of SSI rate increases had at least 2 deficiencies (Table 3).Table 2Poisson regression models comparing surgical site infection (SSI) prevalence rates for procedure clusters receiving statistical process control surveillance to SSI rates for clusters receiving traditional control surveillance.Table 3Compliance with 10 best practices for surgical site infection (SSI) prevention among 502 SSIs analyzed during SSI investigations generated by statistical process control surveillance.SPC methods more frequently detected important SSI rate increases associated with deficiencies in SSI prevention best practices than traditional surveillance; however, feedback of this information did not lead to SSI rate reductions. Further study is indicated to determine the best application of SPC methods to improve adherence to SSI quality measures and prevent SSIs.Arthur W. Baker, MD, MPH, Medincell (Advisor or Review Panel member) Susan S. Huang, MD, MPH, Medline Molnlycke Stryker (Sage) Xttrium"} +{"text": "Indirect calorimetry (IC), which measures oxygen uptake and carbon dioxide output, determines energy expenditure (EE) more precisely than predictive equations in critically ill patients. It is unknown whether the use of IC affects energy provision in critically ill patients with gastrointestinal (GI) conditions that affect absorption and digestionTo (1) compare IC and predictive equations for determining energy needs and (2) evaluate whether IC results affect changes in nutrition support in critically ill patients with GI conditions.In a prospective, observational study, IC was performed for 25 to 55 mins in critically ill patients admitted to intensive care or clinical wards at 2 tertiary-care hospitals in Hamilton, Ontario between Feb 2018 to Sept 2021. EE measured by IC was compared to EE determined by a predictive equation or the Harris-Benedict (HB) formula. A change in energy provision was defined as a change of >10% directed by IC. Continuous data are expressed as means and standard deviation (SD), and categorical data as a proportion of patients. The Mann Whitney U Test (SPSSv26) was used to compare GI and non-GI populations.Of 296 IC tests in 229 patients, 39 of them were in 30 GI patients . Admission GI diagnoses were pancreatitis (33%), liver disease (20%), Crohn\u2019s disease/ autoimmune enteropathy (20%), post-bowel resection (10%), chronic abdominal pain (10%), and cholangitis (7%). The predictive formula underestimated EE in 67% of GI patients compared to IC, corresponding to a mean deficit of 25% of patients\u2019 energy needs. The HB formula underestimated EE in 73% of patients , a mean deficit of 28% of patients\u2019 energy needs compared to IC. Pancreatitis was the majority diagnosis among patients with the highest deficit (>30%) in energy needs when compared to IC. There were no significant differences in the rates of underestimation of energy needs based on predictive and HB formulas between the GI and non-GI patients or between luminal GI and non-luminal GI conditions. After IC, 63% of tests led to changes in energy provisions in GI patients; most requiring an increase in energy provisions (53%).The use of IC to accurately measure EE led to changes in energy provisions in critically ill GI patients. Preventing over- and underfeeding with the implementation of IC to guide nutrition has the potential to improve outcomes in critically ill patients with gastrointestinal conditions.None"} +{"text": "The AgingME Geriatrics Workforce Enhancement Program (GWEP) uses collaboration across institutions of higher education, community-based organizations, and healthcare entities to imbed transformational healthcare practice change across Maine, a primarily rural state. To explore the factors that influence cross-sector collaboration among a diverse array of partners, a baseline anonymous electronic survey was distributed to the newly formed project steering committee. The survey consisted of the Wilder Collaborative Factors Inventory, an established measure of 22 research-based collaboration factors along with four open response questions on process-level challenges and opportunities for improvement. A total of eleven responses (N = 11) were received out of 20 Steering Committee members (55% response). Collaboration strengths noted in the assessment include unique purpose of statewide GWEP efforts (M = 4.41 out of 5 points), mutual trust among members (M = 4.32), favorable social and political environment (M = 4.27), and a history of collaboration among partners (M = 4.27). Lower scores were received on the multiple layers of participation (M = 3.45 out of 5 points), and ability to compromise factors (M = 3.45). Qualitative responses reinforced the need for a common understanding of the project\u2019s goals and outcomes early on in the collaboration. Barriers to collaboration included scheduling considerations and limited time and energy among partners due to heightened COVID-19 response efforts. Results elucidate: 1) Early collaboration strengths and needs of a newly formed statewide education collaborative; and 2) Strategic action steps and focal points informing early partnershipping among organizations engaged in interprofessional health education efforts."} +{"text": "Lung cancer is one of the most widely spread cancers in the world and half of the non-small cell lung cancers are lung adenocarcinoma (LUAD). Although there were several drugs been approved for LUAD therapy, a large portion of LUAD still cannot be effectively treated due to lack of available therapeutic targets. Here, we investigated the oncogenic roles of DKC1 in LUAD and its potential mechanism and explored the possibility of targeting DKC1 for LUAD therapy.DKC1 transcript levels. Gene expression with clinical information from tissue microarray of LUAD were analyzed for associations between DKC1 expression and LUAD prognosis. In addition, loss- and gain-of-function assays were used for oncogenic function of DKC1 both in vitro and in vivo.The Gene Expression Omnibus (GEO) and The Cancer Genome Atlas Program (TCGA) databases were used to examine the DKC1 is positively correlated with telomerase RNA component (TERC) and telomerase reverse transcriptase (TERT) levels in LUAD. DKC1 downregulation resulted in decreased TERC expression, reduced telomerase activity and shorten telomere, and thus eventually led to cell senescence and apoptosis.DKC1 is overexpressed in LUAD compared with adjacent normal tissues. High expression of DKC1 predicts the poor overall survival. DKC1 knockdown in LUAD cell lines induced G1 phase arrest and inhibited cell proliferation. Ectopic expression of DKC1 could rescue the growth of LUAD cell lines. In addition, the abundance of Our results show that high DKC1 expression indicates poor prognosis of LUAD and DKC1 downregulation could induce telomere-related cell senescence and apoptosis. This study suggests that DKC1 could serve as a candidate diagnostic biomarker and therapeutic target for LUAD.The online version contains supplementary material available at 10.1186/s12967-021-02827-0. Lung cancer is the leading cause of cancer death worldwide . Lung adThe dyskerin pseudouridine synthase 1 (DKC1) gene which encodes dyskerin was first identified in dyskeratosis congenita (DC) , 6. DyskThis study aimed to investigate DKC1 expression in LUAD and to examine the relation between DKC1 expression and patient prognosis. Moreover, this study has explored the oncogenic function of DKC1 both in vitro and in vivo and the potential mechanism of DKC1 promoting tumor progression.A549, PC-9 and NCI-H1299 cells were obtained from American Type Culture Collection (ATCC) and maintained by following the culture instructions of ATCC. Human lung adenocarcinoma tissue microarray (HLugA020PG01) was obtained from Shanghai Outdo Biotech Company. There were 84 lung adenocarcinoma tissues and 70 adjacent normal tissues. The clinical data of patients were showed in Additional file DKC1 in 83 LUAD tissues and paired adjacent normal tissues from the Gene Expression Omnibus (GEO) database (GSE75037) was examined. GEO databases (GSE12930 and GSE102016) were used to examine the mRNA level of Dkc1 in inflammatory non-transformed lung. (http://www.ncbi.nlm.nih.gov/geo/). GSE12930 shows gene expression in smoking induced inflammation non-transformed mouse lung P: benzo(a)pyrene, LPS: lipopolysaccharides. ns: no significance. Table S1. The Patients\u2019 clinical data and IHC score from LUAD tissue microarray"} +{"text": "Sarcoptes scabiei mite under 100\u00d7 magnification. Treatment with biseptin (antiseptic lotion) and ivermectin led to a favorable outcome 2 weeks later. Human scabies, a skin infestation caused by Sarcoptes scabiei var. hominis mite, occurs worldwide in all ethnic groups and socioeconomic levels.,Sarcoptes infestation (A 45-year-old woman was referred for foot pain of 3 days duration that impaired walking. Clinical examinations did not reveal any skin abnormalities other than on the plantar aspect of the left foot, where a blister with a linear burrow typical of scabies. Microscopic examination of skin scrapings showed a female estation ."} +{"text": "OBJECTIVES/GOALS: Composition of demographics or image types in publicly available datasets may detract from deep learning (DL) diagnosis performance of underrepresented melanoma subtypes. We evaluate a DL model\u2019s performance on melanoma subtypes that have known association with poor prognosis. METHODS/STUDY POPULATION: We trained a CNN using a single InceptionV3 model for 30 epochs on dermoscopic images of pigmented lesions from the International Skin Imaging Collaboration (ISIC). The ISIC 2018 challenge training set had 10008 total images, with 1113 total nevi, 6705 total melanomas, 97 acral nevi, 10 acral melanomas, 256 head and neck (H&N) nevi, and 164 H&N melanomas. The non-acral test set had 117 melanomas and 200 nevi. The acral test set had 201 melanomas and 161 nevi. The H&N test set had 199 melanomas and 128 nevi. Area under the receiver operating curve (AUC) was calculated. The model was retrained with acral lesion oversampling (10x) and performance on the acral test set was re-evaluated. RESULTS/ANTICIPATED RESULTS: The model performed on the non-acral test with an AUC of 80.5%, on the acral test with an AUC of 76.3%, and on the head and neck test with an AUC of 83.8% After oversampling acral lesions within the training set, the model showed nearly the same performance as without oversampling on acral lesions: AUC of 75.6%. DISCUSSION/SIGNIFICANCE OF IMPACT: Diagnosis of high-risk melanoma subsets remains reliable despite underrepresentation during training, increasing validity for broad implementation of DL technology. Datasets for individual subtypes may not be warranted as findings suggest features may be learned from other skin lesions."} +{"text": "Molecular Biology and Evolution has celebrated papers with powerful impact on our community as reflected by accumulated citations. Below, we highlight ten discoveries, five methods, and five resource publications as \u201cEmerging Classics\u201d based on citations accrued per fractional years since publication. Articles within categories are listed in alphabetical order based on the family name of the first author. Total citation counts were obtained from Web of Science on November 30, 2021. We congratulate these authors on the significance of their contributions and look forward to publishing many high-impact articles in the years to come.Since 2013, Large variation in the ratio of mitochondrial to nuclear mutation rate across animals: implications for genetic diversity and the use of mitochondrial DNA as a molecular marker (2017) Allio and colleagues in Volume 34(11) Pp. 2762\u20132772.Evolution of DNA methylation across insects (2017) Bewick and colleagues in Volume 34(3) Pp. 654\u2013665.The rice paradox: multiple origins but single domestication in Asian rice (2017) Choi and colleagues in Volume 34(4) Pp. 969\u2013979.Deciphering the routes of invasion of Drosophila suzukii by means of ABC random forest (2017) Fraimout and colleagues in Volume 34(4) Pp. 980\u2013996.Between a pod and a hard test: the deep evolution of amoebae (2017) Kang and colleagues in Volume 34(9) Pp. 2258\u20132270.Genomic analysis of European Drosophila melanogaster populations reveals longitudinal structure, continent-wide selection, and previously unknown DNA viruses (2020) Kapun and colleagues in Volume 37(9) Pp. 2661\u20132678.Adaptation of S. cerevisiae to fermented food environments reveals remarkable genome plasticity and the footprints of domestication (2018) Legras and colleagues in Volume 35(7) Pp. 1712\u20131727.Genomic evidence for complex domestication history of the cultivated tomato in Latin America (2020) Razifard and colleagues in Volume 37(4) Pp. 1118\u20131132.Extreme genomic CpG deficiency in SARS-CoV-2 and evasion of host antiviral defense (2020) Xia in Volume 37(9) Pp. 2699\u20132705.Evolution of Rosaceae fruit types based on nuclear phylogeny in the context of geological times and genome duplication (2017) Xiang and colleagues in Volume 34(2) Pp. 262\u2013281.Genomic infectious disease epidemiology in partially sampled and ongoing outbreaks (2017) Didelot and colleagues in Volume 34(4) Pp. 997\u20131007.The unreasonable effectiveness of convolutional neural networks in population genetic inference (2019) Flagel and colleagues in Volume 36(2) Pp. 220\u2013238.A fast likelihood method to reconstruct and visualize ancestral scenarios (2019) Ishikawa and colleagues in Volume 36(9) Pp. 2069\u20132085.New methods to calculate concordance factors for phylogenomic datasets (2020) Minh and colleagues in Volume 37(9) Pp. 2727\u20132733.Theoretical foundation of the RelTime method for estimating divergence times from variable evolutionary rates (2018) Tamura and colleagues in Volume 35(7) Pp. 1770\u20131782.UFBoot2: improving the ultrafast bootstrap approximation (2018) Hoang and colleagues in Volume 35(2) Pp. 518\u2013522.MEGA X: molecular evolutionary genetics analysis across computing platforms (2018) Kumar and colleagues in Volume 35(6) Pp. 1547\u20131549.PartitionFinder 2: new methods for selecting partitioned models of evolution for molecular and morphological phylogenetic analyses (2017) Lanfear and colleagues in Volume 34(3) Pp. 772\u2013773.IQ-TREE 2: new models and efficient methods for phylogenetic inference in the genomic era (2020) Minh and colleagues in Volume 37(5) Pp. 1530\u20131534.DnaSP 6: DNA sequence polymorphism analysis of large data sets (2017) Rozas and colleagues in Volume 34(12) Pp. 3299\u20133302."} +{"text": "Persons may underestimate their risk of HIV infection despite presence of risk factors. Accurate appraisal of HIV risk may assist both patients and providers in preventing HIV acquisition. We evaluated self-perceived risk (SPR) versus calculated risk (CR) of HIV infection in active duty US Air Force (USAF) members with incident HIV infection.USAF members with new HIV diagnosis evaluated at a specialty care military medical center between January 2015-March 2020 with available case report forms were included (n=142). Chart reviews were performed and demographic, social, and clinical characteristics were collected from initial Infectious Disease specialty encounters and case report forms. SPR was characterized as Low or High and compared to CR derived by the Denver HIV Risk Score (DHRS) by points based on patient demographic and risk exposure characteristics.Overall, patients were predominantly male (98%), with a median age of 26 years (IQR 22-30), and the majority (85%) reported same-sex partners (Table 1). Patients more commonly characterized themselves as Low SPR than High SPR . Demographic characteristics were similar except a higher proportion of Low SPR patients (29%) were married or partnered compared to High SPR patients . There was no difference in self-reported condom use (\u226550% of the time) between Low (63%) and High (72%) SPR patients (p=0.28) and documented history of sexually transmitted infections was similarly high in both groups . Previous HIV pre-exposure prophylaxis (PrEP) use was uncommon in both Low SPR (8%) and High SPR (6%) groups. For the evaluation of CR by DHRS (Table 2), both Low and High SPR groups had median scores in the very high risk category (\u226550 points) with similar results by test component.USAF members with incident HIV infection more commonly identified with low SPR despite similar risk behaviors and CRs as high SPR patients. The development of patient education programs and promotion of HIV prevention services including PrEP are needed to reduce incident HIV cases in the USAF. Validated HIV risk calculators like the DHRS may also assist medical providers in identifying candidates for HIV prevention services.All Authors: No reported disclosures"} +{"text": "We aimed to test the effects of motor skill training (MST) on gait automaticity measured by changes in prefrontal cortex (PFC) activation during actual walking. We used data from a 12-week trial of older adults randomized to standard physical therapy and standard+MST in a 1:1 ratio. Functional near infrared spectroscopy (fNIRS) measured PFC activation during simple and dual task walking. We will apply linear mixed models to assess effects of task, time, and MST on PFC activation. We will compare the PFC activation 1) during dual task walking compared to simple walking; 2) across visits after intervention; and 3) between participants receiving MST compared to standard physical therapy. These results will demonstrate whether gait automaticity, as evidenced by PFC activation during walking, is affected by MST."} +{"text": "Physical health monitoring is paramount to optimal care for psychiatric patients. Blood tests and ECGs are invaluable tests throughout a patient's care. At baseline, they aid investigation of potential organic causes of psychiatric presentations and provide organ and electrolyte status before starting medication. Common psychotropic medications carry physical health risks: bloods and ECGs aid in monitoring potential side effects of prescribed medication.In this local Tier 4 inpatient unit, anecdotal observation revealed completion of these basic investigations was noted to be suboptimal.This project aimed to improve timely completion of baseline (within 72 hours of admission) and monitoring (within one week of due date) bloods and ECGS.This project was completed within a 12-bed child and adolescent inpatient unit. Using Plan Do Study Act (PDSA) methodology, the multidisciplinary team collated driver diagrams to identify potential areas for intervention. Following baseline analysis, colleague communication was considered key. Consequently, a chart for bloods and ECG completion was created.PDSA cycle 1: chart implementationPDSA cycle 2: chart simplification and font size increasePDSA cycle 3: allocated change in team leader for this cyclePDSA cycle 4: Blood request pocket in officePDSA cycle 5: chart simplification through removal of datesPDSA cycle 6: ECG pocketPDSA cycle 7: box on handover listEach monthly PDSA cycle included the following consecutive interventions:Monthly investigations and admission numbers are unpredictable and inconsistent in this cohort: relevant case numbers per PDSA ranged from zero to ten. The results were presented as percentages to allow for direct comparison between cycles.Baseline and results of each consecutive PDSA cycle described above were as follows (N/A represents a cycle where no investigations were required):Admission bloods were completed within 72 hours in 50%, 100%, 50%, 80%, N/A, 100%, 100%, 100%Admission ECG was completed within 72 hours in 30%, 66%, 50%, 70%, N/A, 100%, 100%, 100%Monitoring bloods were completed within one week of due date in 25%, 33%, 0%, 80%, 100%, 100%, 100%, 100%Monitoring ECG was completed within one week of due date in 0%, 0%, N/A, 66%, 100%, 66%, N/A 100%Through close multidisciplinary collaboration and chart implementation, completion of bloods and ECGs improved. Low patient numbers per PDSA cycle resulted in large changes in percentage results, limiting the significance of these findings. Wider implementation of the chart within local Trust inpatient wards is considered."} +{"text": "The presence of bifascicular block on electrocardiography suggests that otherwise-unexplained syncope may be due to complete heart block. European Society of Cardiology (ESC) recommends investigating it with electrophysiology study (EPS). PPM is indicated if high-degree atrioventricular block is inducible. Long term rhythm monitoring with implantable loop recorder (ILR) is recommended if EPS is negative. We evaluated adherence to these guidelines.This is a single-center retrospective audit of adult patients with bifascicular block hospitalized for unexplained syncope between January 2018 and August 2019 under general medicine service. Patients with an alternative explanation for syncope were excluded. Guideline adherence was assessed by formal cardiology consult and whether EPS followed by ILR and/or PPM were offered.65 out of 580 adult patients (11.2%) admitted to general medicine service for syncope had a bifascicular block; 29 (5%) were identified to have bifascicular block and unexplained syncope. Median age was 77 \u00b110 years; 9 (31%) were female, and 6 (20.7%) patients had at least one prior hospital visit for syncope at our academic medical center. Cardiology was consulted on 17 (58.6%) patients. Two patients were evaluated by EPS (1 refused) followed by ILR. Overall, 3 out of 29 patients (10.3%) received guideline-directed evaluation during the hospitalization based on ESC guidelines. None of the patients received empiric PPM during the index hospitalization.Among patients admitted to the general medicine service with unexplained syncope and bifascicular block, a minority (10.3%) underwent guideline-directed evaluation per ESC recommendations. Cardiology was consulted in 58.6% of cases. Bifascicular block is defined on electrocardiography as left bundle branch block, right-bundle branch block with left anterior fascicular block , or righWe conducted an audit at our university-based academic tertiary-care hospital to assess adherence to ESC guidelines for adult patients with bifascicular block hospitalized admitted under Internal Medicine service for unexplained syncope.This is a single-center retrospective audit. Electronic medical records of adult patients admitted and discharged by general medicine service with a primary diagnosis of syncope between January 2018 and August 2019 were reviewed. Patients with chronic bifascicular block (confirmed on at least one prior electrocardiogram) and unexplained syncope were identified after thorough chart review including history, physical exam, lab data, electrocardiogram (EKG), echocardiography, and discharge summary. Exclusion criteria were: pre-existing pacemaker; supraventricular or ventricular arrhythmia, or second/third-degree atrioventricular block, bradycardia (heart rate <50 beats per min) with or without the use of negative chronotropic medications; left ventricular ejection fraction < 35%, orthostatic hypotension, vasovagal syncope per history, seizure or recent cerebrovascular accident, cardiac ischemia or infarction related syncope; hypertrophic, infiltrative or inflammatory cardiomyopathy; moderate to severe valvular disease (primary or secondary); abnormal serum magnesium or potassium levels at presentation \u2013patientsGuideline adherence was assessed by formal cardiology consult and whether EPS followed by PPM or ILR were offered. Chi-square test of independence and Fischer\u2019s exact test were performed for statistical analysis.The audit was approved by the institutional review board. There was no patient and public involvement in the design, or conduct, or reporting, or dissemination plans of the research.65 out of 580 consecutive adult patients (11.2%) admitted to general medicine service for syncope had a bifascicular block confirmed on EKG; 29 (5%) were identified to have bifascicular block with no alternative explanation of syncope (unexplained syncope). Fifteen patients (51.7%) had a left bundle branch block. The median age was 77\u00b110 years; 9 (31%) were female, and 6 (20.7%) patients had at least one prior hospital visit for syncope at our academic medical center. The baseline characteristics for the 29 patients with unexplained syncope and bifascicular block are summarized in 1 refusing EPS and opting for ILR + 2 with negative EPS subsequently offered ILR) out of 29 patients (10.3%) with bifascicular block and unexplained syncope received guideline-directed evaluation during the hospitalization based on ESC guidelines (Cardiology was consulted on 17 (58.6%) patients. EPS was offered to 5 patients; 1 refused and opted for ILR directly. EPS was negative in all four patients who underwent testing; 2/4 patients were subsequently offered (and received) ILR and the rest were not offered further treatment. Of the total cohort, 5 patients were directly offered ILR without EPS. Overall, 3 remains unclear and previously unreported, the incidence of unexplained syncope in patients with bifascicular block is estimated to be 5\u20138% , 11. ForOur audit helps demonstrate the value of appropriate evaluation with EPS or ILR\u2013currently being studied in the SPRITELY trial, and highS1 Data(XLSX)Click here for additional data file."} +{"text": "Myxoid liposarcoma (MLS), a malignant soft-tissue tumor derived from lipocytes, is characterized by specific genetic translocations t12;16) and t on a background of few additional chromosomal changes 2;16 and .Previously, we investigated the potential of circulating tumor DNA (ctDNA) to detect tumor recurrence and monitor treatment response . Howevernext generation sequencing (NGS) - based approaches, which allow ultrasensitive detection of MLS DNA in a routine diagnostic setting and prospective clinical trials. As standard NGS panels don\u2019t cover common genetic alterations in MLS, we designed a lockdown panel which encompasses genes with a reported mutation frequency of at least 5%. The 36,541 base pair (bp) standard panel covers the introns of DDIT3, FUS and EWS where the t and t translocations occur, the TERT promoter region and mutation hotspots within exons from seven genes and of 11\u2009bp (SD 73\u2009bp) on chromosome 16 (FUS) with a high intertumor variability on chromosome 12 and exon 20 (c.3140A\u2009>\u2009G) we identified less commonly annotated mutations in exon 5 (c.1035\u2009T\u2009>\u2009A) and exon 8 (c. 1345C\u2009>\u2009A). Only two additional genes were mutated at low frequency in our cohort, TET2 (2%) and PTEN (4%) were additionally quantified by droplet digital PCR. ctDNA levels decreased after tumor resection and increased when metastatic disease was detected. We observed a decline in ctDNA when radio/chemotherapy was initiated. However, with increasing tumor burden, concentrations rose again after several months ctDNA. This indicates that most metastases originated from a clone in the primary tumor, which harbored this mutation allowed us to monitor ctDNA of localized tumors as exemplified in the following scenario. Patient 2 received neoadjuvant radiotherapy and subsequently complete resection (tumor necrosis rate\u2009>\u200990%) of a localized MLS. Two plasma samples obtained at an interval of 9 days before initiation of radiotherapy showed similar concentrations of ctDNA was identified and resected. Exome sequencing was conducted from the initial leg tumor and the lung metastasis. The exome panels targeted 22 and 17 genomic regions respectively, and 6 mutations were identical in both panels. ctDNA in 15 plasma samples was quantified with both panels. During localized disease ctDNA fluctuated depending on the amount of viable tumor mass. The panel from the primary lesion performed superior during localized disease. The small lung metastasis however, led to markedly increased ctDNA concentrations which again decreased after resection. Both panels performed similarly during metastatic disease . Supplementary Figure 3. Comparison of absolute and relative ctDNA quantification. Supplementary Figure 4. Impact of tumor heterogeneity on ctDNA detection. Supplementary Figure 5. Additional target mutations from exome sequencing increase sensitivity of tumor DNA detection. Supplementary Figure 6. Determination of sensitivity and specificity of standard panel.Additional file 2: Materials and Methods."} +{"text": "Ventricular arrhythmias cause a significant proportion of sudden deaths. Several studies demonstrate a high prevalence of ventricular arrhythmias in patients with heart failure regardless of the etiology. The aim of this study was to determine the prevalence of silent ventricular arrhythmias in ambulatory heart failure patients with reduced left ventricular ejection fraction (HFrEF) and its correlation to the prognosis.Four hundred (400) ambulatory HFrEF patients on maximum tolerated doses of heart failure medications were included. Holter monitoring for 7\u00a0days was done in all patients searching for silent ventricular arrhythmias. The patients were followed-up for one year to detect the occurrence of major adverse cardiovascular events. We divided the study population into 2 groups based on an LVEF cutoff value of 30% . Holter monitoring revealed ventricular arrhythmias in 304 patients. Patients with left ventricular ejection fraction (EF)\u2009<\u200930% (Group A) had more complex ventricular arrhythmias in the form of frequent Premature ventricular contractions (PVCs) of\u2009\u2265\u20095% and or non-sustained ventricular tachycardia (NSVT) runs. Furthermore, Among Group A, more major cardiovascular events were observed. Multivariate regression analysis showed that frequent PVCs and severely reduced LVEF were the strongest independent predictors of major cardiovascular events.ventricular arrhythmias are common in HFrEF patients even in the compensated status. Both, left ventricular systolic function and the PVCs burden were found to be the strongest predictors of major adverse cardiovascular events. Heart failure is a clinical syndrome characterized by typical symptoms (fatigue and shortness of breath) that could be associated by clinical signs of systemic or pulmonary venous congestion or both. This is caused by cardiac structural and/or functional abnormality. HF has been divided into three phenotypes based on the of left ventricular ejection fraction (LVEF); heart failure with reduced ejection fraction (HFrEF) with the EF\u2009\u2264\u200940%, heart failure with mildly reduced ejection fraction (HFmrEF) where the EF is from 41\u201349% and heart failure with preserved ejection fraction (HFpEF), with the EF\u2009\u2265\u200950% [Premature ventricular complexes (PVCs) are the most common ventricular arrhythmias. Their impact on the patient's prognosis depends on the presence of cardiac structural and/or functional abnormalities. In the absence of an underlying structural heart disease, PVCs are regarded as a benign phenomenon .Several studies have reported that frequent PVCs could be a trigger for ventricular tachycardia (VT), ventricular fibrillation (VF) and sudden cardiac death (SCD). Moreover, frequent PVCs have been found to cause not only symptoms like palpitations, chest discomfort, and syncope but also contribute to adverse cardiac remodeling and the risk of developing new onset or worsening of heart failure (HF) , 4.Several recent therapeutic approaches, whether pharmacological or device-based, have been introduced into clinical practice to improve the clinical outcome of HF patients. However, the high rate of re\u2010hospitalization and high cardiac mortality are still serious issues .Based on data from a number of studies, almost 80% of patients with reduced left ventricular systolic function have frequent ventricular premature contractions (PVCs), whereas\u2009>\u200940% have runs of non-sustained ventricular tachycardia (NSVT) . SustainThe aim of the present study was to determine the prevalence of silent ventricular arrhythmias on one-week ambulatory ECG monitoring in clinically compensated outpatient heart failure patients with reduced ejection fraction (HFrEF) and its impact on the prognosis .The study included 400 compensated heart failure patients with reduced ejection fraction (EF 25\u201340%) who follow regularly at the outpatient heart failure clinic of Alexandria Main University Hospital and receiving the maximal tolerated doses of beta-blockers, angiotensin converting enzyme inhibitors/Angiotensin II receptor blockers (ACEI/ARBs) and mineralocorticoid receptor antagonists (MRA).Due to financial constraints, no patients were taking angiotensin receptor-neprilysin inhibitors (ARNI).All participants were informed about the study and signed a written consent regarding this study. This study complied with the declaration of Helsinki and was reviewed and approved by the Ethical committee of the Faculty of Medicine, Alexandria University . The diagnosis of HF was determined according to the current HF guidelines .2, abnormal TSH, electrolyte disturbances), current active ischemia , those with cardiac resynchronization therapy (CRT) implantation or treated with class III antiarrhythmic drugs.Patients older than 70\u00a0years, more than mild anemia (<\u200911\u00a0g/dl), evidence of sustained or non-sustained VT on surface ECG, prior cardiac arrest, active metabolic abnormalities , to detect silent ventricular arrhythmias . During Holter monitoring, patients were instructed to do their usual daily activities.Clinical follow up (including 12 lead resting ECG) was performed at 1, 3, 6 and 12\u00a0months intervals. Occurrence of any of the major adverse cardiovascular events was recorded.Data were fed to the computer and analyzed using IBM SPSS software package version 20.0. . Qualitative data were described using number and percent. Quantitative data were described using range (minimum and maximum), mean, standard deviation, median and interquartile range (IQR). Significance of the obtained results was judged at the 5% level.Patients included in this study were predominantly males (93%) with only 7% were females. Their age ranged from 40 to 69\u00a0years. The LV ejection fraction ranged from 25 to 40% with mean value of 32.33\u2009\u00b1\u20094.88. Forty eight percent of the patients had severely reduced LV systolic function with EF\u2009<\u200930% (Group A).The heart rate ranged from 55 to 100\u00a0bpm (mean heart rate was 78\u00a0bpm), the PR interval ranged from 120 to 240\u00a0ms with 36 patients (9%) suffered from first degree heart block, 79 patients (19%) had left bundle branch block (LBBB) and 32 patients (8%) had right bundle branch block (RBBB).Was done in all study patients (as a part of the study protocol) to rule out the presence of significant coronary artery disease, 328 patients (82%) had CAD whether single or multivessel disease.LV systolic function was measured by Simpson-based assessment of LVEF. It was observed that 48% of the patients had severely reduced LV systolic function of\u2009<\u200930% (Group A) and the remaining 52% of the patients had an ejection fraction which ranged from 30 to 39% (Group B).Heart rate ranged from 54 to 120\u00a0bpm.Supraventricular premature contractions were seen in 144 (36%) patients, 40 (10%) patients had short runs of supraventricular tachycardia (SVT) with heart rate reaching up to 170 followed by spontaneous termination.PVCs were recorded in 304 (76%) patients, 108 (27%) patients had infrequent PVCs. 196 (49%) patients had frequent PVCs. All PVCs were monomorphic with no sustained VT recorded on Holter monitoring.No episodes of paroxysmal AF were recorded in any of the study patients.Ischemic cardiomyopathy (ICM) group including 236 patients: infrequent PVCs were detected in 84 patients (35.6%) while frequent PVCs were detected in 152 patients (64.4%).Non-ischemic cardiomyopathy (NICM) group including 68 patients: infrequent PVCs were detected in 24 patients (35.3%) while 44 patients (64.7%) had frequent PVCs.During the follow up period 12\u00a0months), major cardiovascular events were encountered in 272 patients (68%), among which 44 patients (11%) died, 188 patients (47%) suffered from worsening of heart failure symptoms requiring hospitalization and 40 patients (10%) experienced sustained VT with hospital admission. died, 152 patients (70.4%) suffered from worsening of heart failure symptoms with hospital admission while 24 patients (11.1%) experienced sustained ventricular tachycardia with hospital admission.Among the NICM group, 4 patients died (7.1%) while 36 patients (64.3%) had decompensated heart failure which required hospitalization and 16 patients (28.6%) experienced sustained ventricular tachycardia.P value\u2009<\u20090.001). . . Table . There w). Table .Table 5SMultivariate regression analysis of patient characteristics that affect occurrence of major cardiovascular events showed that frequent PVCs (\u2265\u20095% or NSVT) and severely reduced LVEF <\u200930%) are the strongest independent factors that affect major cardiovascular events .This study revealed that PVCs were detected during ambulatory ECG monitoring in 304 patients (76%), among which, 108 patients (27%) had infrequent PVCs and 196 patients (49%) had frequent PVCs . Among the frequent PVCs group, 94 patients (47.95%) had a single or multiple runs of NSVT. Sustained VT was not recorded in any of the studied patients during the ambulatory ECG monitoring period.Regarding the relation of PVC frequency and the underlying substrate, we observed that among the ICM group (the great majority of our study patients), infrequent PVCs were detected in 84 patients (35.6%) while frequent PVCs in 152 patients (64.4%). Among the NICM group, infrequent PVCs were observed in 24 patients (35.3%), while 44 patients (64.7%) had frequent PVCs. PVCs were common finding in HFrEF patients regardless the etiology.Podrid et al. conducteIn our study, we observed that in patients (Group A) with severely reduced LV systolic function (EF\u2009<\u200930%), frequent PVCs were detected in 28 patients (57.1%). While among patients with LVEF\u2009\u2265\u200930% (Group B), frequent PVCs were detected in 21 patients (42.9%).Boas et al. , examineP value\u2009=\u20090.008).Our study showed a statistically significant association between the occurrence of major cardiovascular events and the PVCs burden on Holter monitoring (n\u2009=\u20098) and sustained VT (n\u2009=\u200910). Both age and the admission LVEF predicted lethal VAs on univariate analyses. Receiver operating characteristic curve analysis showed a LVEF cut-off value of 23% predicted the primary endpoint . Furthermore, LVEF at admission independently predicted the primary endpoint on multivariable analysis.Saito et al. eIn contrast, other studies had reported no such relation between VAs and SCD. In one report involving 77 patients with CHF, the presence of NSVT on ambulatory monitoring was more common in those with the greatest degree of LV dysfunction and symptomatic CHF . HoweverTeerlink et al. examinedLip et al. had published an interesting paper on arrhythmias in HF. They reported that VAs were frequently seen as the cause of sudden death or resuscitated sudden death in CHF. Indeed, 50% of all deaths in advanced CHF were sudden and the assumption had been that a significant proportion of these were due to VT/VF. As the severity of CHF increases, the percentage of deaths described as sudden decreases although the absolute risk of VAs and sudden death probably continues to increase .In the current study, a multivariate analysis included the following variables , revealed that LVEF and the PVCS burden were independent predictors of major adverse cardiovascular events .We strongly believe that a lot of research is still needed to understand the predictors of prognosis in heart failure (HF) patients, especially the compensated clinically stable patients, and to better stratify them in order to improve the high morbidity and mortality rates. Our study highlighted the importance of through assessment of such a cohort of asymptomatic HFrEF patients searching for silent arrhythmias.The number of patients enrolled in this study represented a limitation together with the predominance of the male gender and the relatively short follow-up duration.Among heart failure patients with reduced ejection fraction, ventricular arrhythmias are common. Both LV systolic function and the burden of PVCs are regarded as the strongest predictors of major adverse cardiovascular events."} +{"text": "Recombination-mediated cassette exchange (RMCE) is a recently developed alternative method for creating single copy transgenes using recombination rather than repair of double stranded breaks as the mechanism for driving integration into the genome. Two alternative methods for performing RMCE have been developed; a two-component approach using an unlinked source of FLP recombinase, and a one-component approach using a FLP expression cassette within the landing site. Here, I describe new landing sites for performing both types of RMCE. The new landing sites are located within 50 bp of well-vetted MosSCI insertion sites on Chr II and Chr IV. C. elegans. The recent development of recombination-mediated cassette exchange (RMCE) integration approaches in worms provides a relatively rapid efficient method to create single copy transgenes with inserts up to at least 12 kb . RMCE is similar to MosSCI in that it depends on integrating at landing sites that have been engineered. In the case of MosSCI, a transposon is excised, and the double stranded break is repaired by a template often using synthesis-dependent strand annealing (SDSA) which is error prone . Similar approaches mediated by cas9 cleavage also have high error rates, approaching 65% in some studies . By contrast, RMCE uses recombination to insert the template into the genome which rarely yield erroneous inserts .Transgenic animals are powerful tools in the study of basic biological processes using FRT and FRT3. Recombination between the FRT and FRT3 sites in the plasmid and the landing site yields replacement of the genomic FRT FRT3 interval with plasmid FRT FRT3 interval. This recombination likely occurs in two steps; first a loop in by recombination at one of the sites, followed by excision by recombination at the other. These recombination events typically occur in the F1 germline, though occasionally it occurs in the P0 animal .The RMCE approach I developed takes advantage of two distinct recombinases. A plasmid template delivered into the gonad of young adult animals is first integrated into the genome using FLP recombinase. Both the template and the landing site contain two distinct FLP integration sites, bqSi711). Injection of the plasmid leads to integration of the plasmid at the landing site which is identified as a Rol (or HygR) animal. After the initial insertion is made homozygous, the self-excising marker cassette (SEC) is then excised using a heat shock Cre protocol, leading to the final insertion. The insertion is then outcrossed from bqSi711 using simple crosses. The second method utilizes a landing site which contains a germline FLP expressing transcription unit contained within the landing site. In this case, the FLP expression element is excised using a heat shock Cre protocol. One limitation of the RMCE approach is the lack of landing sites for integration. Here, I describe several new landing sites created using a CRISPR integration approach.Two methods have been developed for performing RMCE. The first method uses a landing site and an unlinked source of FLP recombinase . I also integrated a single-component landing site at same position on Chr II and another at a site adjacent to cxTi10882, another commonly used Mos1 insertion site on Chr IV. In addition, I modified the previously described two-component landing site jsTi1453 I, deleting the left miniMosarm from that landing site . I first characterized these novel landing sites by integrating the identical mec-4p GFP-C1 tbb-2 3\u2019 construct at each site and comparing the expression level of insertions at the new landing sites to identical insertions at the previously described landing sites . The expression of GFP-C1 in touch receptor neurons (TRN) was easily detected in all the new landing site transgenes, though the expression level was slightly lower than that observed in integrations at the four previously described sites .I integrated a two-component landing site using CRISPR/cas9 just adjacent to the position of the tetO/tetR Tet OFF system exhibit background expression in both the pharynx and the rectal gland cells . Because a similar rectal gland background signal was observed at several different landing sites using distinct bipartite systems, I speculated that the miniMos transposon arm might be contributing to the background. Comparison of the identical tetO 7X \u2206mec-7p GFP-C1 reporter integrated at both jsTi1453 and the js1570 \u2206mosL arm derivative confirmed this was the case as expression in the rectal gland cells was undetectable in the js1570 derived transgene . Despite the reduction in background, the tetO reporter still robustly expressed GFP in TRNs, when driven by the identical mec-4 promoter tet OFF driver .Recently developed bipartite reporters including a https://sites.wustl.edu/nonetlab/rmhe/), I have used these new landing sites to create addition RMCE insertions. I collated the insertion frequency data from a set of over 90 injection sessions in which I counted the number of Roller F1 animals obtained, and the number of insertions obtained . These data demonstrate that js1570, jsSi1579 and jsSi1669 all behave comparably to previously described landing sites, yielding insertions at a rate of approximately 1 per 3 injected animals. However, the jsSi1691 single component site yielded insertions at a lower frequency . I speculate this is due to lower expression of FLP from the jsSi1691 landing site since integration at the same position using jsSi1579 and bqSi711 as a source of FLP yields normal integration frequencies. A recent study indicated that for CRISPR/cas9 genome modifications one could obtain a much higher frequency of integration events using specifically treated miniprep DNA . A preliminary set of 7 injections performed while this manuscript was under review and presented in In developing a new recombination-mediated homolog exchange technique . I have also characterized most of the transgenes obtained at these additional landing sites either by confirming the presence of an expected fluorescence pattern, recombinase activity, The new landing site are now available at the CGC and should provide additional flexibility in creating RMCE-based transgenic animals. I also plan to create additional landing sites at well-characterized high expressing genomic positions on the remaining chromosomes that currently do not contain landing sites.C. elegans was maintained on NGM agar plates spotted with OP50 at 22.5\u00b0C or at 25\u00b0C during the RMCE protocol.RMCE transgenesishttps://sites.wustl.edu/nonetlab/rmhe-vectors/) and injected at ~50 ng/\u00b5l into young adults. In the set of injections summarized in et al. (2021) and injected at 40-50 ng/ul. Integrants were identified and isolated as described in detail in Nonet (2020). Performing RMCE at 25\u00b0C is critical to obtaining robust integration rates. The criterion for including an injection session in the table was obtaining a mean of at least 1 F1 Rol per injected animal. All injection sessions into js1570, jsSi1669 and jsSi1691 met this criterion. Eleven jsSi1579 injection sessions failed to meet this criterion. They consisted of 5 sessions injecting plasmids with strong ubiquitous promoters (eft-3 or rpl-27) driving tet OFF and 6 sessions injecting plasmids that contained both a tet OFF driver and a tetO reporter cassette. In all cases dead eggs were observed on the injection plates. In cases where a fluorescent protein reporter was in the plasmid, the dead eggs were brightly fluorescent. The two jsSi1579 failures included in the table were one rpl-27 session and one dual driver and reporter plasmid session. In some cases, I was able to integrate plasmids containing both a driver and a reporter by injecting animals growing on doxycycline.Inserts were cloned into pLF3FShC , pRMHEB or pRMHEP (CRISPR/cas9-mediated insertions and deletionjs1570 was created using a dpy-10 co-CRISPR strategy. A mix of plasmids NMp3143 (40 ng/ul), NMp3153 (10 ng/ul), NMp3828 (20 ng/ul), NMp3829 (20 ng/ul) and oligonucleotides NMo5238 (0.4 uM) and NMo6761 (0.4 uM) we co-injected into jsTi1453; bqSi711 animals. Rol progeny were screen by PCR (NMo6564/6569) for presence of the deletion and homozygosed. The deletion structure was confirmed by sequence analysis. jsSi1579 was created by injecting unc-119(ed3); bqSi711 animals with a mix of NMp3143 (40ng/ul), NMp3630 (50 ng/ul), NM3631 (30 ng/ul). Insertions were identified by selection for hygR Rol progeny by adding 25 ul of 100 mg/ml HydroGold\u2122(InvivoGen) to P0 injection plates 3 days after injection. After isolating homozygotes, the R sqt-1hyg self-excision cassette (SEC) in the insertion was excised by screening for non-Rol progeny after a 20-hr. heat shock at 29\u00b0 C. The structure of the insertion was validated by a combination of long-range PCR [performed as outlined in Nonet (2020) using MNo3887/3888], restriction digestion and sequence analysis. jsSi1691 was created by injection of N2 (the wild type) with a mixture of plasmids NMp3143 (40 ng/ul), NMp3630 (50 ng/ul), NMp4043 (20 ng/ul), pBluescript KS (50 ng/ul), pGH8 (2 ng/ul) and pCFJ90 (2.5 ng/ul). Insertions were selected for and analyzed as described for jsSi1579. jsSi1669 was created by injection of oxTi1127 animals with a mixture of plasmid NM4055 (35 ng/ul), NM4057 (25 ng/ul), pBluescript KS (+) (50 ng/ul), pGH8 (2 ng/ul) and pCFJ90 (2.5 ng/ul). Insertions were identified by hygromycin selection as described above. The structure of the insertion was validated as described above using oligonucleotides NMo3889/3890. The genomic sequence of the insertions is available at https://sites.wustl.edu/nonetlab/rmce-insertion-strains/ .Microscopyet al., 2012) using a 40X air lens at 20% LED power with 100 ms exposures. PLM soma and ALM soma signals were quantified using the FIJI version of ImageJ software as described in Nonet (2020). Rectal gland cell images presented were collected using the same conditions.For quantification of GFP signals, homozygous L4 hermaphrodite animals were mounted on 2% agar pads in a 2 \u00b5l drop of 1mM levamisole in phosphate buffered saline, cover slipped and imaged on an Olympus BX-60 microscope equipped with a Qimaging Retiga EXi monochrome CCD camera, a Lumencor AURA LED light source, Semrock GFP-3035B and mCherry-A-000 filter sets, and a Tofra focus drive, run using micro-manager 2.0\u00df software (Schindelin Plasmid constructionsIntegration vectors were assembled using Golden Gate (GG) reactions as described in Nonet (2020). Other plasmids were constructed using standard cloning techniques.The following previously published plasmids were used:et al., 1988), pDD162 , pCFJ90 and pGH8 , and NMp3055, NMp3421, NMp3467, NMp3470, NMp3631, NMp3643, NMp3732, NMp3746 and NMp3774 .pBluescript KS (+) (Short The following plasmids were constructed:NMp3143 peft-3-cas9 (3 int)A derivative of the pDD162 Cas9 expression plasmid lacking the empty U6 sgRNA cassette. pDD162 was amplified using NMo5228/5379 and re-circularized using a Gibson assembly reaction.NMp3150 DR274 U6 FEU6 promoter sgRNA clone with a flipped and extend sgRNA as described in Ward (2015). sgFE was amplified from NMp3055 using NMo5407/5075, purified, digested with BamHI and HindIII, and inserted into BamHI and HindIII digested NMp3055.NMp3153 DR274 U6 FE dpy-10dpy-10 at GCTACCATAGGCACCACgAG. NMp3150 was digested with BsaI and the annealed oligonucleotide pair NMo5236/5237 was inserted by ligation.U6 promoter sgRNA targeting NMp3630 DR274 U6 MosIIttTi5605Mos1 insertion site at gatatcagtctgtttcgtaa cgg. NMp3055 was digested with BsaI and the annealed oligonucleotide pair NMo6439/6440 was inserted by ligation.U6 promoter sgRNA targeting Chr II adjacent to the NMp3828 DR274 U6 sgTi1453 FjsTi1453 landing site at attcacggcacaacatacat tgg. NMp3055 was digested with BsaI and the annealed oligonucleotide pair NMo6757/6758 was inserted by ligation.U6 promoter sgRNA targeting Chr I adjacent to the NMp3829 DR274 U6 sgMosLminiMos adjacent to the left arm at gttgAGCTCCACCGCGGTGG CGG. NMp3055 was digested with BsaI and the annealed oligonucleotide pair NMo6759/6760 was inserted by ligation.U6 promoter sgRNA targeting NMp4043 pSAP ChrII FLP loxP FRT FRT3 landingChr II full RMCE landing site CRISPR template. The left arm amplified from N2 genomic DNA using NMo6707/6450, the right arm amplified from N2 genomic DNA using NM06708/6453, and the loxP FLP FRT FRT landing site from NMp3746 were co assembled into NMp3421 using a SapI Golden Gate assembly reaction.NMp4053 DR274 5\u2032 arm cxTi10882 left armcxTi10882. The left arm was amplified from N2 genomic DNA using NMo7060/7061 and inserted into NMp3467 using a BsaI Golden Gate reaction.Left arm genomic fragment adjacent to NMp4054 DR274 3\u2032 arm cxTi10882 right armcxTi10882. The right arm was amplified from N2 genomic DNA using NMo7058/7059 and inserted into NMp3470 using a BsaI Golden Gate reaction.Right arm genomic fragment adjacent to NMp4055 DR274 U6 cxTi10882cxTi10882 Mos1 insertion site at actgttggatgcctgtgtag cgg. NMp3055 was digested with BsaI and the annealed oligonucleotide pair NMo7062/7063 was inserted by ligation.U6 promoter sgRNA targeting Chr IV adjacent to the NMp4057 pSAP cxTi10882 FLP loxP FRT FRT3 landingChr IV full RMCE landing site CRISPR template. The left chr IV arm from NMp4053, the right Chr IV arm from NMp4054 and the loxP FLP FRT FRT landing site from NMp3746 were co assembled into NMp3421 using a SapI Golden Gate assembly reaction.OligonucleotidesNovel transgenesWorm StrainsCaenorhabditis Genetics Center (CGC). Plasmids and additional strains and will be submitted to Addgene or CGC and if demand levels warrant it.Plasmids are available by request from MLN. Strains containing the four new landing sites have been submitted to the"} +{"text": "Dear Editors,Randomised control trial (RCT) evidence is not available to guide screening for asymptomatic coronary artery disease before kidney transplantation . United Responses were received from all 23 (100%) centres, of which 22 had a protocol for cardiac assessment prior to listing. In three centres, asymptomatic individuals were not required to undergo cardiac investigation beyond an ECG or echocardiogram prior to transplantation. The remainder followed a risk-stratified approach; no centres performed universal screening.In centres adopting risk-stratified screening, factors used to screen patients included a history of ischaemic heart disease (100% of centres), diabetes (100%), peripheral vascular disease (50%), smoking (50%), stroke 35%), limited exercise capacity (35%), hyper/hypotension (15%), or an abnormal echocardiogram (95%) or ECG (70%). Two centres stratified using the Newcastle Risk Index [5%, limitThe most frequent screening investigation was a myocardial perfusion scan (55%) followed by stress echocardiogram (20%). Coronary angiography and cardiopulmonary exercise testing were the initial investigation in one centre each. Other indications for coronary angiography included an abnormal initial screening test (39%) or on cardiology advice (35%). In one third of centres, the waiting time for investigations was over 10\u00a0weeks.Nine centres had cardio-renal multidisciplinary meetings, whilst 14 had a designated cardiologist providing transplant candidate assessments. In 16 centres cardiology review was only needed for patients with abnormal screening tests, whilst in three cardiologists reviewed all screened patients.Of 23 centres, 10 had updated their screening protocol within the past 2\u00a0years and three were in the process of an update. Whilst 19 centres reported experience of patient declines from listing based on an abnormal screening test, this amounted to one patient per month or less in 11 centres.Respondents commented on the challenges of outdated evidence, reliance on observational data, and differences between real-world cohorts and RCT study populations when assessing the evidence for cardiac screening. The importance of cardio-renal meetings was noted in units not adopting screening. Of 23 centres, 22 expressed interest to participate in an RCT to examine the utility of screening, 12 of whom supported recruiting the highest cardiac risk candidates.Our survey highlights variation in screening practice across the United Kingdom . Similar"} +{"text": "It is unclear based on published literature whether shorter courses of antibiotic treatment may be appropriate for urinary tract infections (UTI) in patients with SCI/D given their complex baseline clinical status. C. difficile infection or death within 30 or 90 days, respectively, after treatment completion. Statistical tests included Chi-square, Mann-Whitney U, and logistic regression. This retrospective cohort study was conducted at the VA San Diego Healthcare System (VASDHS), which has a dedicated SCI/D unit. Adults with SCI/D were identified for inclusion if they had received antibiotics for a positive urine culture in conjunction with UTI symptoms from 1/2018-12/2020. Individual UTI events were excluded if associated with potential sources of harbored infection, anatomic abnormalities increasing risk of bacteriuria, non-bacterial pathogens, concurrent infections prolonging antibiotic treatment, or antibiotic courses managed outside of VASDHS. Treatment groups comprised UTI events treated with no more than 7 days of antibiotics (group 1) versus more than 7 days (group 2). Study endpoints were recurrence or new incidence of UTI within 30 and 90 days after completion of antibiotic treatment and onset of C. difficile infection occurred in 1 patient in group 2 after 3 days of antibiotic therapy. Duration of antibiotic therapy was not predictive of treatment failure within 30 days of antibiotic completion. Factors predictive of treatment with longer courses of antibiotic therapy included hospital admission and more severe ASIA impairment scale score. One-hundred and seven patients with 241 unique UTI events were included in this study, with 79 events in group 1 and 162 events in group 2. Baseline characteristics were similar across both groups, aside from a higher incidence of hospital admission and more severe SCI/D based on the American Spinal Cord Injury Association (ASIA) impairment scale in group 2. Efficacy outcomes are described in Table 1. No deaths occurred within 90 days of treatment completion, and Table 1. Incidence of UTI after antibiotic completionThe findings of this study suggest that for some patients with SCI/D, UTI treatment lasting 7 days or fewer may be effective compared to longer courses of antibiotics and could be beneficial in reducing collateral damage from antibiotic use. All Authors: No reported disclosures"} +{"text": "ABSTRACT IMPACT: Recommendations for increasing trainee productivity will be highlighted. OBJECTIVES/GOALS: Using a combination of qualitative (interview) and quantitative (publications tracking) data, we undertook to describe the hurdles and concerns impeding academic accomplishments among T and K awardees at one CTSA hub and to examine whether hurdles at 6 months would predict academic output within one year following completion of the training program. METHODS/STUDY POPULATION: Semi-structured interviews were conducted with 29 trainees (28 TL1 and 8 KL2) 6 months into their training. Interview transcripts were analyzed using Atlas.ti to identify hurdles and concerns . PubMed searches yielded the number of publications within one year of exiting the training program. Frequencies of hurdles and concerns were examined to characterize the factors most likely to impact trainee progress during the first 6 months of their training program. Among 18 trainees who had completed their training, the mean number of publications within one year of exiting the program (identified via verified PubMed searches) was compared across the total number of hurdles reported at 6 months (range = 0 to 3). RESULTS/ANTICIPATED RESULTS: The thematic analysis yielded 19 categories of hurdles and 14 categories of concerns. The top three hurdles were technological challenges , professional competing responsibilities (40%), and navigating collaborations (30%). The top three concerns were future funding (33%), potential as an independent researcher (27%), and institutional context . The number of hurdles reported at 6 months significantly predicted number of publications one year post-exit = 3.14, p < .05). Trainees reporting zero hurdles generated a mean of 8.67 publications; those with 3 hurdles generated a mean of 2.4 publications. DISCUSSION/SIGNIFICANCE OF FINDINGS: Future concerns were completely different from past hurdles, suggesting that the issues impeding research progress are not anticipated. Results suggest trainees would benefit from training related to how to balance competing professional responsibilities and navigate collaborations and that early attention to hurdles may enhance productivity."} +{"text": "Malignant meningiomas comprise 2\u20135% of all meningiomas. The process of malignant transformation when benign meningiomas (WHO grade I-II) become malignant (WHO grade III) has not previously been investigated in sequential tumour surgeries. Upregulation of FOXM1 expression and DREAM-complex repression have shown phenotypical subgroups correlating with WHO grade and aggressiveness. We investigated the RNA expression of 30 genes central to meningioma biology and 770 genes involved in neuroinflammatory pathways in primary and secondary malignant meningioma patients who underwent one to several operations.We identified a cohort of consecutive malignant meningioma patients treated at Rigshospitalet, Copenhagen from 2000\u20132020 (n=51) and gathered their malignant tumours and previous WHO grade I/II tumours. The malignant cohort (MC) was counter matched with a benign cohort (BC) where patients had no recurrences during follow-up. RNA expression signatures from 140 samples from the MC and 51 samples from the BC were analysed with the Nanostring Neuroinflammation panel customized with 30 genes known to be relevant in meningioma phenotypes.49% of MC patients had a previous grade I/II meningioma making them secondary malignant meningioma patients. Progression-free survival calculated from first malignant surgery to first recurrence or death showed no significant difference in the primary vs. secondary patients. Preliminary results of single-gene analysis of MC tumours showed FOXM1, MYBL2, TOP2A, BIRC5 expression was higher in WHO grade III samples. Gene-expression signatures in the individual patients and gene ontology enrichment analyses are in process.FOXM1, MYBL2, TOP2A, BIRC5 RNA expression levels seem to rise during malignant progression across patients. Gene-expression analysis using the Nanostring technology is feasible and a potentially powerful tool to distinguish meningiomas prone to malignant transformation from truly benign meningiomas."} +{"text": "Scarring is a major outcome of severe burn wound healing. Severe scars often persist and diminish quality of life by disfigurement, pain, and pruritis. In the last decade, utilization of ablative fractional carbon dioxide (CO2) laser therapy has become a popular treatment modality for severe burn scars. Although the efficacy of CO2 lasers for the treatment of hypertrophic burn scars has been established via systematic reviews, there have been no attempts to query the 63 American Burn Association (ABA) centers across the United States regarding specific treatment parameters involving serious, sometimes high, total body surface area (TBSA) burns.Throughout October and November of 2020, a Qualtrics survey consisting of 14 questions was administered to burn surgeons practicing at all 63 ABA burn centers across the United States. Topics assessed were specific laser parameters utilized (5), treatment preferences (2), peri-operative follow-up (5), scar assessment practices (1), and TBSA treatment tolerance (1).Exploratory, descriptive data was analyzed. Surgeons practicing at 27 of the 63 total ABA burn centers responded to the survey (43% response rate). Data elucidates the level of variance regarding current initial management of hypertrophic burn scars via CO2 laser treatment. Surgeons demonstrated variation in the level of TBSA treatment tolerance and pulse energy settings, respectively.Our findings show a substantial amount of variation in several aspects of CO2 laser hypertrophic scar revision between ABA centers across the country, including preoperative evaluation, laser settings, treatment regimen, and postoperative recommendations . Standardization of care when utilizing ablative fractional CO2 lasers should be further explored."} +{"text": "Due to the lack of institutional support, families have long been the primary caregivers in China. Most studies to date only focused on one single care activity during a particular life course stage. Nonetheless, older adults today are more likely to care for multiple family members concurrently or sequentially . The studies on discrete snapshots of care activities failed to capture the patterns of family caregiving overtime. Utilizing four waves of longitudinal data from CHARLS , this study particularly focuses on care activities to grandchildren, parents, and spouse, and maps out the family caregiving patterns overtime. Using latent profile analysis, this study identifies five family caregiving patterns: 1). Light grandchild caregivers (27%), who on average provided 4.3 years\u2019 grandchild care mostly; 2). Heavy grandchild caregivers (11%), who on average on provided 7 years\u2019 grandchild care mostly; 3). Light caregivers for grandchildren and parents (7%), who sequentially provided 1-year care to grandchildren and parents; 4). Heavy serial caregiver (6%), who mostly provided care to spouse and grandchildren with higher overlapping years; 5). Overall light caregivers (49%), who on average provided less than one year of care to any recipient. The preliminary results suggest that heavy serial caregivers (6%) far worst in terms of depressive symptoms and more likely to report worsened self-rated health; and overall light caregivers (49%) have the lowest depressive symptoms and more likely to report good self-rated health."} +{"text": "Unexpected donor-derived hepatitis B virus (HBV) infection is defined as a new HBV infection in a recipient of a transplanted organ from a donor who tested negative for total antihepatitis B core antibody , hepatitis B surface antigen (HBsAg), and HBV DNAAll suspected unexpected cases of donor\u2010derived hepatitis B in the United States are reported to the Organ Procurement and Transplantation Network for review by the Ad Hoc Disease Transmission Advisory Committee. Suspected cases are referred to CDC to investigate whether donor-derived disease transmission occurred and identify interventions to prevent transmission and improve outcomes , or false-positive total anti-HBc (two) or HBsAg (two) results. Twenty confirmed cases were included.Median age at death of the 20 donors was 31 years (range = 20\u201346 years); 11 were male, and 19 were White. The most common cause of death was drug intoxication. Injection drug use and positive toxicology were each reported for 18 donors . SixteenNew HBV infection was identified in 18 liver and two liver-kidney recipients at a median of 41 weeks after transplantation (range = 5\u2013116 weeks). Among cases reported during 2019, hepatitis B test conversion was first identified at a median of 38 weeks after transplantation . None ofHBV infection among transplant recipients can occur from reactivation of previous HBV infection before transplantation and HBV DNA testing at 4\u20136 weeks after transplantation (Unexpected donor-derived hepatitis B virus (HBV) infection after organ transplantation is rare and is associated most commonly with donor injection drug use.During 2019, the Organ Procurement and Transplantation Network and CDC received an increased number of reports of HBV infection among liver recipients from HBV-negative donors; 12 of 14 implicated donors had evidence of recent injection drug use, and 13 donors were hepatitis C virus (HCV)\u2013seropositive.Providers caring for recipients of organs from donors who are HCV\u2013seropositive or who recently injected drugs should maintain awareness of infectious complications of drug use and monitor recipients accordingly."} +{"text": "Recent data supports that improved qualitative antibody responses correlate with elite controllers (EC) of HIV. As ADCC has been associated with protection in vaccine studies, thorough exploration of antibodies that facilitate ADCC is warranted. In studies on monoclonal antibodies from long-term non-progressors (LTNPs), our laboratory has previously described highly mutated antibodies against a complex conformational epitope with contributions from both gp41 heptad repeat regions. Despite using the VH1-02 gene segment, known to contribute to some of the broadest neutralizing antibodies against HIV, members of these antibodies, termed group 76C antibodies, did not exhibit broad neutralization. Our goal was to characterize the non-neutralizing functions of antibodies of group 76C, to assess targeting of the epitope in various clinical presentations, and to assess the development of these antibodies by comparison to their predicted common ancestor. Serum samples were obtained from HIV+ clinical groups: EC, LTNP, stable CD4 counts on therapy, and those off therapy.In antibody/serum competition assays, comparison to VRC01 which also uses VH1-02, showed that antibodies targeting the 76C group epitope were enriched in LTNPs. We then show recombinant antibodies of 76C members 6F5 and 6F11 both have robust ADCC activity, despite their sequence disparity. Sequence analysis predicted the common ancestor of this clonal group would utilize the germline non-mutated variable gene. We produced a recombinant ancestor Ab (76Canc) with a heavy chain utilizing the germline variable gene sequence paired to the 6F5 light chain. 76Canc binds HIV envelope constructs near the original group C epitope. 76Canc also shows comparable ADCC to 6F5 and 6F11 on both clade B and C constructs.Common ancestor antibodies maintain function and these types of antibodies correlate to a non-progressive clinical state.(A) Serum from long-term non-progressors (LTNPs) compared to serum from a group of HIV infected with lower CD4 levels as a control for viral load were used to compete against biotinylated CD4 binding site (VRC01) and 76C Gp41 conformational epitope (6F11) targeting antibodies. Serum dilutions were chosen to align means near 50%. Means with 95% confidence intervals are shown. (B) Monoclonal antibody 76Canc was created using the germline sequence of the heavy chain variable region with the CDR3 and light chain of 76C member. Antibody dependent cell cytotoxicity flow cytometric based assays were performed using gp41 proteins from clade B (MN) and clade C (ZA1197).Certain antibodies present early on in infection may contribute to overall clinical course. Variable gene germline sequences that support functional activity against HIV could be targeted in vaccine regimens.All Authors: No reported disclosures"} +{"text": "Alzheimer's disease and related dementias (ADRD) are a significant public health burden. Preventing hospitalizations in adults with ADRD is a public health priority. Data from the 2016\u20132018 Healthcare Cost Utilization Project National Inpatient Sample, an all-payer representative sample of US hospitalizations, were used to describe potentially preventable hospitalizations in adults \u226545 years with ADRD using International Classification of Disease, Tenth Edition, Clinical Modification (ICD-10-CM) codes. Definitions for principal or any-listed ICD-10-CM codes from the Agency for Healthcare Research and Quality defined potentially preventable hospitalizations where admissions might have been avoided by appropriate outpatient primary care management. Of discharges in adults \u226545 years with a potentially preventable hospitalization diagnosis, 11.4% had a diagnosis of ADRD listed in any position. Of those discharges with ADRD, a significantly higher proportion (82.6%) with diagnosis related to potentially preventable hospitalizations were aged \u226575 years compared to 78.9% without potentially preventable hospitalizations. Additionally, of those with ADRD and potentially preventable hospitalization diagnoses, a higher proportion died in the hospital (5.7%) compared to those without potentially preventable hospitalization diagnoses (3.4%). The most common potentially preventable hospitalization diagnoses among adults with ADRD were related to sepsis (34.0%), injuries (20.8%), urinary tract infections (14.2%), and heart failure (12.7%). Measures focusing on preventing injuries as well as identifying early signs and symptoms of potentially preventable hospitalizations like urinary tract infections and sepsis in adults with ADRD could reduce the number of preventable hospitalizations in this population."} +{"text": "The relationship between ERCC gene polymorphism and osteosarcoma risk / overall survival of osteosarcoma is still conflicting, and this meta-analysis was performed to assess these associations.The association studies were identified from PubMed, and eligible reports were included and calculated using meta-analysis method.Four studies were included for the association of ERCC gene polymorphism with osteosarcoma risk, and nine studies were recruited into this meta-analysis for the relationship between ERCC gene polymorphism and overall survival of osteosarcoma. The meta-analysis indicated that ERCC1 rs3212986 (8092 C>A) gene polymorphism, ERCC1 rs11615 (19007 T>C) gene polymorphism, ERCC2 rs1799793 (A>G) gene polymorphism, ERCC2 rs13181 (Lys751Gln) gene polymorphism were not associated with osteosarcoma risk. ERCC1 rs2298881 (C>A) gene polymorphism, ERCC1 rs3212986 (8092 C>A) gene polymorphism, ERCC1 rs11615 (19007 T>C) gene polymorphism, ERCC2 rs1799793 (Asp312Asn) gene polymorphism were not associated with overall survival of osteosarcoma. Interestingly, ERCC2 rs13181 A allele and GG genotype were associated with overall survival of osteosarcoma, but AA genotype not .ERCC2 rs13181 A allele and GG genotype were associated with overall survival of osteosarcoma. Human osteosarcoma, one of the most familiar forms of the primary malignant tumor to adolescents and adults, is a genetically heterogeneous bone malignancy with poor prognosis despite the employment of aggressive chemotherapy regimens Within DNA repair genes, there lie a number of single nucleotide polymorphisms which may impair protein function and attenuate DNA repair capability, resulting in genomic instability and individual predisposition to malignancies The search was conducted in the databases of PubMed on May 1, 2019, and the relevant investigation were included. The retrieval strategy of \u201c(ERCC2 OR ERCC3 OR ERCC1 OR excision repair cross-complementing) AND (osteosarcoma OR bone tumour OR bone cancer OR bone carcinoma) AND polymorphism\u201d was entered into the PubMed database.Inclusion criteria: (1) The outcome must be osteosarcoma or overall survival of osteosarcoma; (2) The study included two comparison groups (case group vs control group); (3) report should give the data of ERCC genotype distribution.Exclusion criteria: (1) Case reports, editorials and review articles; (2) Preliminary result not on ERCC gene polymorphism or osteosarcoma / overall survival of osteosarcoma; (3) Investigating the role ERCC gene expression to overall survival of osteosarcoma.The following information from each recruited investigation was extracted by two investigators independently (Guanliang Wang and Jianping Li): first author's surname, year of publication, ethnicity, control source of the control group and the number of cases and controls for ERCC genotypes. Frequencies of allele of ERCC were calculated for case group and control group. Quality assessment was assessed by Newcastle-Ottawa Scale (NOS) score, and it was regarded as a high quality (or low-bias risk) study when total stars achieved six to nine.Cochrane Review Manager Version 5 was used in this meta-analysis to count the extracted data from each report. The pooled statistic was counted using the fixed effects model. However, a random effects model was conducted when the P value of heterogeneity test was less than 0.1. Results were expressed using odds ratios (OR) for dichotomous data. 95% confidence intervals (CI) were also calculated. P < 0.05 was required for the pooled OR to be statistically significant, and I2 was used to test the heterogeneity among the included studies.The database of Pubmed was searched for this meta-analysis, and 11 studies were eligible and included for this meta-analysis, and the recruited flowchart is shown in Two studies Two studies Two studies Three studies Two studies Four studies Eight studies Seven studies Seven studies This meta-analysis was performed to detect the relationship between ERCC gene polymorphism and osteosarcomas risk, and the relationship between ERCC gene polymorphism and overall survival of osteosarcoma. We found that ERCC1 rs3212986 (8092 C>A) gene polymorphism, ERCC1 rs11615 (19007 T>C) gene polymorphism, ERCC2 rs1799793 (A>G) gene polymorphism, ERCC2 rs13181 (Lys751Gln) gene polymorphism were not associated with osteosarcoma risk. ERCC1 rs2298881 (C>A) gene polymorphism, ERCC1 rs3212986 (8092 C>A) gene polymorphism, ERCC1 rs11615 (19007 T>C) gene polymorphism, ERCC2 rs1799793 (Asp312Asn) gene polymorphism were not associated with overall survival of osteosarcoma. Interestingly, ERCC2 rs13181 A allele and GG genotype were associated with overall survival of osteosarcoma, but AA genotype not.In previous, Li et al ERCC2 rs13181 A allele and GG genotype were associated with overall survival of osteosarcoma. However, more association investigations are required to confirm these associations."} +{"text": "Mycobacterium tuberculosis (Mtb) has complex and dynamic interactions with the human host, and subpopulations of Mtb that emerge during infection can influence disease outcomes. This study implicates zinc ion (Zn2+) availability as a likely driver of bacterial phenotypic heterogeneity in vivo. Zn2+ sequestration is part of \u201cnutritional immunity\u201d, where the immune system limits micronutrients to control pathogen growth, but this defense mechanism seems to be ineffective in controlling Mtb infection. Nonetheless, Zn2+-limitation is an environmental cue sensed by Mtb, as calprotectin triggers the zinc uptake regulator (Zur) regulon response in vitro and co-localizes with Zn2+-limited Mtb in vivo. Prolonged Zn2+ limitation leads to numerous physiological changes in vitro, including differential expression of certain antigens, alterations in lipid metabolism and distinct cell surface morphology. Furthermore, Mtb enduring limited Zn2+ employ defensive measures to fight oxidative stress, by increasing expression of proteins involved in DNA repair and antioxidant activity, including well described virulence factors KatG and AhpC, along with altered utilization of redox cofactors. Here, we propose a model in which prolonged Zn2+ limitation defines a population of Mtb with anticipatory adaptations against impending immune attack, based on the evidence that Zn2+-limited Mtb are more resistant to oxidative stress and exhibit increased survival and induce more severe pulmonary granulomas in mice. Considering that extracellular Mtb may transit through the Zn2+-limited caseum before infecting na\u00efve immune cells or upon host-to-host transmission, the resulting phenotypic heterogeneity driven by varied Zn2+ availability likely plays a key role during early interactions with host cells. Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), has plagued humanity for millennia and remains the world\u2019s deadliest bacterium today. Bacterial heterogeneity is one of the most important characteristics of Mtb that complicates TB treatment. Access to zinc ion (Zn2+) may influence bacterial heterogeneity, considering microenvironments developed during TB create a perpetual cycle exposing Mtb to high and low concentrations of Zn2+. Here we show that Zn2+ limitation drives changes in gene expression patterns of well described virulence factors in Mtb, and Zn2+-limited Mtb show increased resistance to oxidative stress and increased replication in vivo. Our results suggest that host-pathogen interactions are influenced by pre-exposure of Mtb to Zn2+ and mycobacteria that transit through a Zn2+-depleted microenvironment are primed to withstand impending oxidative stress upon subsequent contact with immune cells in the same, or a na\u00efve host. Considering that the standard mycobacterial media recapitulates a Zn2+-replete environment, the Zn2+-dependent phenotype of the pathogen may confound our fundamental understanding of initial interactions between Mtb and immune cells. Mycobacterium tuberculosis (Mtb) as a human pathogen is enabled by a genetic arsenal that allows it to withstand a myriad of immune defenses, survive in diverse host environments, and establish persistent infection are experienced by Mtb throughout infection. Zn2+ rapidly accumulates in phagosomes inside macrophages and low [Zn2+] in CP-rich caseum. The spectrum of diverse immunopathologies of granulomas and necrotic cavities give rise to distinct subpopulations of Mtb that exist simultaneously within the host [2+] may be a contributing factor in their development. Further, [Zn2+]-dependent physiological changes may affect interactions with the immune system, thus influencing disease progression and response to treatment.One cue that triggers physiological adaptation in many pathogens is the limitation of essential micronutrients, mmunity\u2019 . Indeed,agosomes , but Mtbar niche . Infectear niche . As TB dne cells ,10. Necrthe host ,5, and may likewise delineate subpopulations of Mtb in vivo. We use biochemical and multi-omics approaches to describe the effect of prolonged Zn2+ limitation at a global scale in Mtb grown in vitro. The analysis of Zn2+-limited Mtb revealed a response that goes beyond the Zur regulon, including activation of the oxidative stress response, altered utilization of reducing cofactors, changes in the lipidome and a distinct cell surface morphology. In addition, Zn2+-limited Mtb are more resistant to oxidative stress and more sensitive to the prodrug isoniazid compared to Zn2+-replete Mtb. Finally, in an aerosol mouse model of infection, Zn2+-limited inoculum exhibited greater bacterial burden and pulmonary granulomas than Zn2+-replete inoculum. Together these findings define a novel adaptive mechanism employed by Mtb during Zn2+ limitation that triggers formation of a distinct population that may play a role in TB pathogenesis.In this study, we show that Mtb transmission into the airways and out of the host via sputum explains the largest portion of variation in the data (nearly 80%), with intra-sample variation having a much smaller effect and alkylhydroperoxide reductase (AhpC) and many genes involved in DNA replication, repair and response to DNA damage . NAD(P)H FLIM-phasor technique has previously been used to differentiate metabolic states in live bacterial populations at the single-cell level on anlot, Mtb 2+] on anin vivo before being transmitted to na\u00efve hosts, it is relevant to determine if [Zn2+] alone can affect virulence of Mtb. To test this, we used C3HeB/FeJ (Kramnik) mice which develop liquefied necrotic granulomas resembling human disease pathology influences a myriad of changes at the gene level correlating with changes at the protein level and translating into altered physiological characteristics of [Zn2+]-derived populations of Mtb. Many Zn2+ binding proteins have essential biological functions during infection and have adapted elaborate mechanisms to endure Zn2+ toxicity and Zn2+ limitation in vivo is likely a physiologically relevant signal experienced by Mtb in vivo, since we were able to demonstrate that this micronutrient triggers the formation of distinct populations in vitro. Zn2+-limited Mtb exhibit a global adaptive response that affects physiology, confers resiliency to oxidative stress and possibly leads to increased virulence. Mtb depends on a cycle of exit and re-entry into host immune cells to perpetuate its lifecycle, and [Zn2+] may be a major cue experienced in this cycle that could potentially affect host-pathogen interactions and disease outcome .In agreement with our observations and consistent with the notion that ZnA repair ,56,57. Ig agents , furthern (furA) , isocitr) , is the best characterized and appreciated property of the tubercle necrotic granuloma in preparation for transit through the hypoxic caseum. The complete lack of DosR-mediated hypoxic response in Zn2+-limited Mtb demonstrates that [Zn2+] alone does not recapitulate the microenvironment of sputum or the necrotic granuloma. We suggest that, while hypoxia is a relevant cue to bacilli in certain microenvironments during tuberculosis, [Zn2+] may be another cue that delineates bacterial physiology in vivo, possibly even independent of hypoxia, and we emphasize that the combined effects of hypoxia and Zn2+ limitation remains obscure.Hypoxia, not alone dramatically contributes to changes in mycobacterial physiology. We discovered that Zn2+-limited bacteria have increased resistance to exogenous oxidation as predicted by increased antioxidant expression in this condition. While Zn2+-limitation did not have a broad effect on antibiotic susceptibility, Zn2+-limited bacteria were more resistant to ROS-producing rifampicin while increased KatG expression sensitized this subpopulation to the prodrug isoniazid , including upregulation of numerous transcription factors and enzymes involved in the oxidative stress response leading to a global adaptive response during Zn2+ limitation. Accordingly, predisposition of individual Mtb bacilli to the Zn2+-limited microenvironment could prime them to interact differentially with the host during infection used in this study were approved by the Texas A&M University Institutional Animal Care and Use Committee. Sputum leftovers from TB testing were obtained without identifiers from Hawaii Department of Health and were not considered human subject study. Detailed materials and methods are provided in S1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file.S3 Fig(PDF)Click here for additional data file.S4 Fig(PDF)Click here for additional data file.S5 Fig(PDF)Click here for additional data file.S6 Fig(PDF)Click here for additional data file.S7 Fig(PDF)Click here for additional data file.S8 Fig(PDF)Click here for additional data file.S9 Fig(PDF)Click here for additional data file.S10 Fig(PDF)Click here for additional data file.S11 Fig(PDF)Click here for additional data file.S12 Fig(PDF)Click here for additional data file.S13 Fig(PDF)Click here for additional data file.S14 Fig(PDF)Click here for additional data file.S15 Fig(PDF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S7 Table(XLSX)Click here for additional data file.S8 Table(XLSX)Click here for additional data file.S9 Table(XLSX)Click here for additional data file.S10 Table(XLSX)Click here for additional data file.S1 Data(XLSX)Click here for additional data file.S1 Text(PDF)Click here for additional data file."} +{"text": "Nontuberculous mycobacteria (NTM) infection is associated with high rates of morbidity and mortality among immunocompromised adults. However, sparse data exists regarding clinical outcomes among immunocompromised (IC) children with NTM infection. We sought to characterize clinical features and outcomes among IC children at our institution with microbiologically confirmed NTM disease.Retrospective review of cases of microbiologically confirmed NTM infection among IC children between January 2017 and December 2020. Children (\u226421y.o) with microbiologically confirmed NTM disease and known primary or secondary immunodeficiency diagnosed between January 1, 2017 and December 20, 2020 were included in the study. All subjects with a positive NTM microbiologic stain or culture but no subsequent treatment for NTM infection were excluded. Demographic and clinical characteristics were assessed and risk factors for mortality were evaluated.M. abscessus (50%) and M. avium complex (30%) the most frequently implicated pathogens. Hospital acquired infection was common (60%). 2 year mortality following invasive NTM infection was high at 40%. Of 147 mycobacterial cultures sent during the study period, 72 subjects had a positive microbiologically confirmed NTM species, with 10 subjects meeting all inclusion and no exclusion criteria. Median age was 16 years old, with 40 percent being female and 50 percent of Hispanic ethnicity. NTM disease was distributed among patients with primary immunodeficiency (30%), solid organ transplantation (20%), hematopoietic stem cell transplant (20%), rheumatologic disease on immunosuppressive therapy (10%), and hematologic malignancy (10%). Bacteremia was common, with blood cultures positive in 70% of cases, and While rare, NTM infections are associated with significant morbidity and mortality among immunocompromised children. Additional investigations are needed to assess for risk factors associated with NTM and severe NTM disease.Laura Filkins, PhD, Avsana Labs Biofire Diagnostics (Grant/Research Support)"} +{"text": "Adverse childhood experiences (ACEs) have long-term impacts on health throughout the life course. Emerging research found that 3+ ACEs are associated with increased risk of cognitive impairment, nearly 11 times more than those who have not experienced childhood adversity. This study further investigates the ACEs-SCD relationship using data from the 2011 Behavioral Risk Factor Surveillance System (BRFSS). Seven ACE questions were asked of respondents in California, Washington, and Wisconsin ; SCD was measured as experiencing progressive confusion or memory loss in the last 12 months. A series of logistic regressions were run to separately model the presence of ACEs and ACE score on SCD. Fourteen percent reported SCD, with 65.4% of those reporting 1+ ACE. More SCD respondents reported 4+ ACEs (10.8%) than non-SCD respondents (4.8%). The most frequently reported ACEs among those with SCD were psychological abuse (34.9%) and substance abuse in the household (30.5%). Regression results showed greater SCD risk with increased ACE scores, up to 2.90 odds of SCD for 4+ ACEs compared to 0 ACEs (p<.0001). Those reporting physical abuse and sexual abuse had the greatest odds of SCD. Findings demonstrate a strong association between childhood adversity and SCD, with physical and sexual abuse placing individuals at greatest risk. Results show possible pathways to which ACEs can lead to cognitive impairment. Findings implicate the importance of considering a lifespan perspective in childhood adversity and family violence work and the importance of considering early-life adversity when assessing risk for cognitive impairment."} +{"text": "The correct title is: Mathematical modelling and control of African animal trypanosomosis with interacting populations in West Africa\u2014Could biting flies be important in maintaining the disease endemicity? The correct citation is: Odeniran PO, Onifade AA, MacLeod ET, Ademola IO, Alderton S, Welburn SC (2020) Mathematical modelling and control of African animal trypanosomosis with interacting populations in West Africa\u2014Could biting flies be important in maintaining the disease endemicity? PLoS ONE 15(11): e0242435."} +{"text": "OBJECTIVES/GOALS: Acute lung Injury (ALI) has long been considered a proceeding event to the development of Acute Respiratory Distress Syndrome (ARDS). Diagnosis of classical ALI and ARDS remains difficult relies on clinical components of the Berlin Criteria, interpretation of radiographs and exclusion of pulmonary edema inducing processes. The precipitating factor for developing ALI involves direct or indirect insult to the lungs. Recent studies have described metalloproteinase-3 (MMP3) to be elevated in plasma samples of patients with lung injury and potentially affected by tobacco use. MMP3 can degrade extracellular matrix components contributing to lung edema and inflammation. This study was conducted to examine the utility of matrix metalloproteinase-3 (MMP3) as a biomarker of lung injury. METHODS/STUDY POPULATION: We conducted a single center, retrospective cohort study of patients admitted to the medical ICU (MICU). De-identified bronchoalveolar fluid (BALF) samples were collected and stored at \u221280C. Enzymatic activity of MMP3 was determined using a fluorescent resonance energy transfer (FRET) assay. Demographics, comorbidities, evidence of lung injury and patient outcomes were collected. Data were reported with descriptive statistics and data was analyzed with t-tests for statistical significance. RESULTS/ANTICIPATED RESULTS: 55 patient BALF samples were included in the final analysis . 54.5% (n = 30) of patients were determined to have lung injury, 29% (n = 16) of patients had COPD and 45.5% (n = 25) of patients were smokers. MMP3 was higher in patients with lung injury . Smoking was associated with decreased MMP3 activity . COPD was not associated with differences in MMP3 . DISCUSSION/SIGNIFICANCE OF IMPACT: Lung Injury results in elevated MMP3 levels. Smoking was not shown to increase MMP3 levels and may in fact increase them. COPD demonstrated no effect on MMP3 levels. MMP3 levels may vary based on the mode of lung injury (i.e. direct vs indirect) and smoking may impact the activity of the enzyme. Further research should assess activity of MMP3 through different modes of lung injury."} +{"text": "DMH1 treated mice had a modest decrease in trabecular bone and reduced lymphocytes in circulation without affecting tumor growth. Taken together we show unique responses to BMP inhibition in metastatic prostate cancer in the bone. These studies suggest that profiling bone lesions in metastatic prostate cancer can help identify therapeutic targets that not only treat the metastatic tumor but also address the need to better treat the distinct tumor induced bone disease.From the 33,000 men in the U.S. who die from prostate cancer each year, the majority of these patients exhibit metastatic disease with bone being the most common site of metastasis. Prostate cancer bone metastases are commonly blastic, exhibiting new growth of unhealthy sclerotic bone, which can cause painful skeletal related events. Patient\u2019s current care entails androgen deprivation therapy, anti-resorptive agents, radiation, and chemotherapy to help control the spread of the cancer but little intervention is available to treat blastic bone disease. The transforming growth factor beta (TGF\u03b2) and bone morphogenetic protein (BMP) pathways are known to regulate bone growth and resorption of destructive lytic bone lesions, yet the role of TGF\u03b2/BMP signaling in prostate cancer blastic vs lytic bone lesions are not fully understood. We hypothesized that to target the BMP/TGF\u03b2 pathway, a useful biomarker of bone lytic or blastic pathology would have superior response. We show distinct BMP vs. TGF\u03b2 signaling in clinical samples of human prostate cancer bone metastases with either lytic or blastic pathologies. BMPs exhibit distinct effects on bone homeostasis, so to examine the effect of BMP inhibition on healthy bone, we treated mice with the BMP receptor small molecule antagonist DMH1 and saw a modest temporary improvement in bone health, with increased trabecular bone. We next sought to use the BMP inhibitor DMH1 to treat bone metastasis engraftment seeded by a caudal artery injection of the lytic human prostate cell line PC3 in immunodeficient mice. The colonization by PC3 cells to the bone were restricted with DMH1 treatment and bone health was importantly preserved. We next proceeded to test BMP inhibition in an injury model of established bone metastasis Tumor induced bone disease (TIBD) continues to present as a high morbidity causation in the progression of many metastatic cancers . Metastain vitro to enhance matrix deposition or promote osteogenesis programs, both of which can be reversed with BMP inhibition obtained from the University of Colorado Pathology Shared Resource. Ex vivo mouse tissues were harvested and immediately placed in 10% formalin and fixed for 24 hours. Formalin was replaced with 14% EDTA for 5-7 days until decalcified and placed in 70% ethanol for 24 hours prior to embedding in paraffin wax. FFPE tissue blocks were sectioned at 5\u00b5m thickness with two sections per slide and mounted on plus coated microscope slides. Slides were dewaxed and rehydrated with the following sequence of washes: xylene (StatLab), 100% ethanol, 95% ethanol, 70% ethanol, 50% ethanol, and then phosphate buffered saline (PBS) . Heat-induced antigen retrieval was performed by microwaving slides in citrate pH6 Antigen Unmasking Solution (Vector Laboratories) in a rice cooker. Routine H&E staining was performed in Harris hematoxylin (Vector Labs). Primary antibodies for pSMAD1/5/8 (Millipore 1:100) and pSmad3 (Abcam 1:200) were used for immunohistochemistry (IHC) staining. Signal was detected by ImmPRESS polymer secondaries to appropriate host and DAB chromogen substrate (Vector Labs) and counterstained with Hematoxylin QS (Vector Labs). All bright field IHC and H&E were scanned at 40X (0.22um/pixel) magnification using a ScanScope XT System (Aperio Technologies). To quantitate IHC staining, a grid of up to five 20X images per slide was captured, avoiding excessive stroma or necrotic tissues, on an Eclipse Ni microscope (Nikon) and imported into ImageJ to auto-contrast images then change contrast to blue and then the threshold was set and staining was measured by percent area.via IP injection (25ug/100\u00b5l). Animals were anesthetized using isoflurane 2-5%, site was prepped for a small incision at the base of the shoulders for subcutaneous implantation of the pump. Skin was closed using wound clips (BD 427631). 6 and 12 weeks from initial treatment the animals were analyzed by Dual-energy X-ray absorptiometry (DXA) while anesthetized. At week six, with pump expired, the ZA groups received an additional injection of zoledronic acid. To model bone metastasis in mice, a Caudal Artery Injection (CAI) of PC3 cells in 100ul sterile PBS were injected into the tail via the caudal artery as previously reported (5 MyC-CaP cells in 10\u00b5L PBS were injected into the tibia of FVBn (Jax Stock # 001800) male mice no younger than 8 weeks of age. Mice were bred and maintained at Rocky Mountain Regional Veterans Affairs Medical Center (protocol number CD1611M) as well as at the University of Colorado Anschutz Medical Campus (protocol number 00553). All the immune competent mice (FVBn and C57bl6) were housed together and immunodeficient animals (NSG) were house solitarily because of aggression in accordance with ARRIVE guidelines males at 8 weeks of age with healthy appearance and weights over 20g were used. Mice were placed into four groups (n=5/group). Mice received a 6-week osmotic pump (Alzet 2006 #0007223), experiments in reported . NSG was performed weekly or biweekly while animals were under anesthesia of inhaled isoflurane of 2-5%. Analysis of the femurs to quantify bone mineral content (BMC) and density (BMD) were performed by manually drawn region of interest surrounding the femur or tibia bilaterally under the bone only option. Bones were scanned with a \u00b5CT scanner at 70kV, 114mA with a 0.5mm aluminum filter using a pixel size of 6\u00b5m. Images were acquired at 0.6\u00fe through 180\u00fe, reconstructions and analysis were performed using NRecon, DataViewer and CTAn software (Bruker). ROI for trabecular bone , 2mm were analyzed 0.1 mm distal to the growth. For cortical tibial midshaft ROI; 2mm proximally from the tibia/fibula junction, 1mm was analyzed. IVIS Spectrum (Perkin Elmer) was used to image the animals with RediJect 2-DeoxyGlucosone (DG) 750 (Perkin Elmer 760561). IVIS imaging of 750nm 2-DG was performed as directed by using the wizard setting with Ex Filter 745 and Em Filter 800 .The human prostate cancer cell line PC3 cell line was obtained from the American Type Culture Collection (ATCC) and cultured in DMEM (Cat#10-013-CV) Medium (Corning) 1X Antibiotic-Antimycotic (Thermofisher Scientific Cat#15240062) and 10% Fetal Bovine Serum (FBS) (Seradigm). The FVBn MYC-CaP cell line was obtained from ATCC/Dr. Austin Kirschner at Vanderbilt University and cultured in DMEM (Corning), Antibiotic-Antimycotic and 10% FBS (Seradigm). All cell lines were routinely tested for mycoplasma infection by PCR and authenticated by morphology and published growth rates available from ATCC. Additional DNA fingerprint authentication services were performed the CU Cancer Center Tissue Culture Core Facility for the PC3 cell line.Submandibular blood collection was performed prior to sacrifice, 200\u00b5l of blood was drawn using lancets (Goldenrod) . Blood wStatistical analyses were performed using GraphPad Prism and Excel . All statistical tests used a cutoff p-value of 0.05 for significance with the Mann-Whitney test . Multiple group comparisons were analyzed by one way ANOVA in GraphPad with a Kruskal-Wallis post-test. The choice for these statistical tests were to compare significant differences in numerical values originating from similar sample types such as bone radiologic data.To investigate whether canonical readouts of BMP and TGF\u03b2 signaling is altered in TIBD we performed Immunohistochemistry (IHC) on clinical samples containing bone with metastatic prostate cancer. These samples were previously utilized to identify cellular and molecular differences in the tumor microenvironment of TIBD based on their bone pathology, either being enriched for lytic destructive bone lesions or sclerotic and blastic bone features . Staininvia intraperitoneal (IP) injection. Mice underwent DXA X-ray at six weeks and representative images of femurs revealed no deficiencies in bone or a control DMSO pump alone or in addition to the anti-resorptive bisphosphonate Zoledronic acid (ZA) via the tail caudal artery. The next day mice were given an IP injection of Zoledronic acid and subcutaneous implant of a 4 week duration osmotic pump carrying DMSO (n=4) or DMH1 30ug/ml (n=5) then were euthanized upon pump expiration after 4 weeks elevated in DMH1 treated tumors, potentially highlighting the efficacy of DMH1 in vivo and PBS was injected into the right tibia as a control as well as at the University of Colorado Anschutz Medical Campus (protocol number 00553). All animal procedures were performed in accordance with the National Institutes of Health\u2019s Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committees.PO conceived and designed experiments. DS, CI, MP, and PO performed experiments. DS and PO wrote and edited the manuscript. All authors contributed to the article and approved the submitted version.This work was supported by VA Grant 1KBX00002929 (PO) and NIH grant P30CA046934 for the Colorado Cancer Center Support grant.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Drug development in recurrent glioblastoma multiforme (rGBM) is challenging. For randomized controlled trials (RCTs) short survival horizons and limited life-prolonging treatment options may delay accrual and introduce bias through differential dropout of control patients. Comparing results of a single-arm Phase 2b trial of intratumoral delivery of MDNA55 (an interleukin-4 receptor targeted fusion protein) to an external control arm, we sought early efficacy insights and consideration by the FDA of incorporating an ECA in a Phase 3 registrational trial.Using propensity score weighting, we compared rGBM patients from the Phase 2b trial (NCT02858895) (2017-2019) to patients from rGBM registries who had received standard of care therapies (2011-2019) and met eligibility requirements. Propensity scores were estimated using a logistic regression model with 11 covariates. We compared the propensity score weighted groups according to demographic and disease attributes before and after weighting and compared overall survival between the two groups.Through propensity score weighting, 43 MDNA55 patients and 40.80 weighted ECA patients (from 62 unweighted registry patients) were identified for comparison. MDNA55 and ECA patients were balanced on all baseline characteristics . Compared to ECA patients, MDNA55 patients had a 37% lower hazard of death .In advance of a Phase 3 trial, comparison of Phase 2b trial results to an ECA suggests that MDNA55 may be efficacious in rGBM. In view of the known challenges associated with drug development for rGBM, these results provided a proof-of-concept for the design of a novel hybrid Phase 3 trial. This planned Phase 3 trial incorporates propensity score weighting to create a composite hybrid randomized and external control arm, an approach preferred by the FDA over full replacement of a randomized control with an external control."} +{"text": "During the COVID-19 pandemic, many older adults were not receiving primary care services because they could not negotiate the technology for telehealth visits. Coupled with persisting pandemic physical distancing, increased social isolation in older adults was- and continues to be a significant problem. To combat these issues, we aimed to 1) prepare older adults for longitudinal isolation by encouraging social connectedness, and 2) enable older adults to safely access remote primary care services during the pandemic. We paired older adults from 9 housing sites in Los Angeles, CA with health professions graduate students from 9 programs at USC (N = 88 dyads) and provided iPhones to participants without a smartphone. Students educated and supported older adults about the use of technology to access primary care services and to socially connect with family/friends. When requested, 3 additional students provided enhanced 1:1 technology support. Among the 45 participating older adults who received iPhones (51.1%), 22 requests were made for enhanced technology assistance during the 6-month program. Most requests related to initial setup/navigation of iPhone (81.8%) or video calls (27.3%), where others requested help with Wi-Fi (13.6%), composing emails (4.5%), and adding language/translation features (4.5%). Nineteen (83%) technology support requests were successfully resolved; the remaining were unresolved due to loss to follow-up. Our findings demonstrate that older adults can successfully cross the digital divide when technology support is provided. Additionally, pairing older adults with health professions students is an effective strategy to enable access to remote primary care and social connectedness."} +{"text": "Water adsorption and the progressive conversion of vapor-phasegrown oxide particles into hydroxides give rise to either one-dimensionalor two-dimensional (1D or 2D) structures of high dispersion and surfacearea. The resulting Mg(OH)2 lamella with a predominant(001) surface termination are well-suited precursor structures fortheir topotactic conversion into laterally extended and uniform MgO(111)grain surface configurations. To understand the potential of polar(111) surfaces for faceting and surface reconstruction effects associatedwith water desorption, we investigated the stability of MgO(111) nanosheetsduring vacuum annealing and electron beam exposure. The significantsurface reconstruction of the MgO(111) surfaces observed shows thatadsorbate-free (111)-terminated surfaces of unsupported MgO nanostructuresreconstruct rather than remain as charged planes of either three-foldcoordinated O2\u2013 ion or Mg2+ ions. Thus,here we demonstrate the role water can play in surface formation andreconstruction by bridging wet chemical and surface science inspiredapproaches.Developing simple,inexpensive, and environmentally benign approachesto integrate morphologically well-defined nanoscale building blocksinto larger high surface area materials is a key challenge in materialsdesign and processing. In this work, we investigate the fundamentalsurface phenomena between MgO and water (both adsorption and desorption)with particles prepared via a vapor-phase process (MgO nanocubes)and a modified aerogel process (MgO(111) nanosheets). Through thesestudies, we unravel a strategy to assemble individual MgO nanoparticlesinto extended faceted single-crystalline MgO nanosheets and nanorodswith well-defined exposed surfaces and edges. This reorganizationcan be triggered by the presence of H The organization of MgO nanocubes asprimary building blockscan be achieved by materials contact with water and subsequent thermaltreatment in a vacuum. It is well establishedthat the size, shape, and faceting of metaloxide particles have a substantial impact on their catalytic activityas well as their chemical properties when utilized as catalyst supports.Realizing cost-effective and environmentally benign approaches tocontrollably incorporate metal oxide nanoparticles into extended structureswith well-defined faceting would be a significant step in progressingmaterials development across numerous fields of research and industrialapplications. One promising approach, presented herein, is the topotacticrearrangement of metal oxide nanoparticles after their exposure toaqueous conditions.5 In that regard, particles with characteristic and defined crystalhabits are promising building units to generate spatially organizednanostructures composed of uniformly sized and shaped surface elements.As a result of their simple morphology and the limited number of characteristiclocal surface structures, cubic metal oxide particles with a rocksalt structure representparticularly well-suited model compounds to study such phenomena.The transformation behavior of metal oxideparticles in aqueousenvironments is characterized by a variety of interaction pathwaysthat include oriented attachment and dissolution\u2013recrystallizationprocesses.6 flame spray pyrolysis (FSP),7 or simply the combustion of metallic magnesium in air.6 When these MgO particles are synthesized at hightemperatures and in anhydrous oxygen atmosphere, they adopt a typicalcubic morphology, bound by the thermodynamically most stable surfaces.8 Such particles have well-defined crystallinity,crystallite habit, particle size distribution,9 andhost a high abundance of characteristic surface features such as corners,edges, and step edges as well as other defects which can be identifiedby different structural characterizations and spectroscopic12 techniques. In addition, it has been demonstrated that water adsorptionand surface hydroxylation have a strong influence on the surface morphologyof MgO grains.13 Ab-initio calculationshave shown that the order of surface energies of the different (100),(110), and (111) planes is reversed as a consequence of the strongeradsorption (chemisorption) energy of water to the high index planes.15 Protonation stabilizes the otherwise unstable (111)surface, which exhibits the highest stability under ambient conditions.17 Thus, cubic MgO is well-suited to study processes that rely on interfacialand growth phenomena within metal oxide particles when exposed toan aqueous environment.18This is especially true for MgO particles produced by gas phasesynthesis techniques such as chemical vapor synthesis (CVS),19 and gas sorption.20 The rock salt (111) surface is classified as a Tasker \u201ctype3\u201d surface with alternating layers of cations and anions ,36 and they demonstrate properties significantly different than thoseof the more typical (100) and (110) surfaces of the same material,38 with enhanced activity being attributed to under-coordinated sitesat corners and step terraces, as well as exposed O2\u2013 sites at point defects. Recently, it was reported that MgO(111)treated at 800 \u00b0C demonstrated a 65% increase in CO2 capacity despite suffering from a 30% decrease in surface area dueto sintering. It was established that the heat treatment removed surfacehydrogen exposing the low-coordinated O2\u2013 necessaryfor CO2 sorption.20 In otherrecent work, Gates and Richards et al. succeeded in the uniform anchoringof iridium atoms over the edge and corner elements of such facetedMgO(111) microplanes to generate precise and periodic structures ofatomically dispersed metals on a crystalline high surface area support.39The surfacestructure, and hence chemistry, of metal oxides isin large part dependent on the oxide facet. Of particular interestis the (111) facet of MgO, which has demonstrated increased activityfor varying applications including heterogeneous catalysisd anions 1. Accord40 Herein, we report on extended faceted single-crystalline MgO nanosheetsand nanorods with well-defined exposed surfaces and edges synthesizedvia topotactic decomposition of sheets of hydroxylated MgO and Mg(OH)2. Such structures are of great interest because of the uniformdistribution of their corner and edge elements and high degree of(111) faceting.To take advantage ofsuch surface-induced effects, a variety ofsolution-based syntheses have been developed to produce sheet-likeMgO nanostructures with exposed {111} planes.2O vapor or in contact with condensed water to generate characteristic1D structures and 2D faceted surfaces. Vacuum annealing experimentsat 273, 473, and 1173 K lead to different degrees of faceting, whichcan be explained by the different adsorbate coverages resulting fromthese different annealing temperatures. Additionally, we address theeffect of reconstruction and faceting41 that occur at MgO(111) surfaces to compensate for the energeticallyunfavorable surface polarity that arises in the final stages of dehydroxylationand dehydration. Adsorbate removal can be achieved under high vacuumannealing at T > 1073 K or during electron beamirradiationwithin a transmission electron microscope. The transformation stepsdescribed here are simple and describe the conversion of nanocubesrandomly organized within dry powders into regular stacks of nanocubesthat are interconnected via the cube edges. The conversion processexclusively involves MgO nanoparticles, gaseous, or liquid water atdefined dosages and subsequent vacuum treatment. Thus, we report arobust and simple route for the synthesis of ultrathin and thermallystable high surface area supports, which are in high-demand for numerousapplications including heterogeneous catalysis. In the second partof this study, we discuss wet-chemistry derived MgO (111) nanosheets,stabilized by residual synthesis related adsorbates together withtheir structural derivatives that emerge upon vacuum annealing at T > 1073 K or extended electron beam exposure. Our resultsshow that bare MgO(111) reconstructs into the more stable (100) surfaces,41 suggesting that completely bare polar MgO(111)surfaces do not exist.In the first part of this study, we report onthe structural reorganizationpathways that MgO nanocubes can undergo in the presence of H7 Stable process conditions areguaranteed by spatially separating the evaporation and oxidation zone.The synthesis reactor consists of two quartz glass tubes inside acylindrical furnace. The inner tube hosts ceramic ships with Mg pieces, which are heated to 913 K assuring a metal vaporpressure of 1 mmHg column (1.33 mbar). An inert argon stream carriesthe metal vapor away from the evaporation zone to the end of the innerglass tube. There the Ar/metal vapor mixture encounters the oxidizingagent (O2), which is flowing through the outer glass tube.The exothermic oxidation reaction leads to a bright stable flame inthe reactor, and MgO nanoparticles form because of the homogeneousnucleation and crystal growth in the gas phase. Thanks to continuouspumping, the residence time of nuclei within the flame remains shortenough to prevent substantial coarsening and coalescence. A bypasssystem allows avoiding particle collection during uncontrolled processconditions, i.e., the heating and cooling phase. The total pressurein the CVS reactor is kept constant at 50 \u00b1 3 mbar over the entireproduction process.For the production of MgOnanocubes, we use chemical vapor synthesis, which allows for the controlledevaporation and subsequent oxidation of alkaline earth metals underreduced pressures.\u20131 andexposure to molecular oxygen at this temperature. Then, the sampletemperature was raised to 1173 K at pressures p <5 \u00d7 10\u20136 mbar and kept at this temperaturefor 1 h until full dehydroxylation of the sample surface was achieved.42After the gas phase synthesis, the MgO nanoparticlepowders are transferred into quartz glass cells, which allow one tocarry out thermal activation of the powders in defined gas atmospheres.The as-obtained MgO powders are cleaned of organic contaminants byheating to 1123 K at a rate of 5 K\u00b7min2 uptakefrom the atmosphere could contribute to the here observed transformations,we also performed Ar flushing for convective mixing of the dispersion.Finally, the dispersion is centrifuged, and the solid material isdried using a membrane pump (p < 2 mbar) for 24h prior to materials characterization.After vacuum annealing, 50 mg of the MgOpowder is dispersed in100 mL of high-grade water (Millipore Simplicity M 185). The dispersionis stirred for 30 min on a magnetic stirrer. Alternatively and forcontrol experiments that should rule out that CO35 4-Methoxybenzyl alcohol is added as a directing agent and is hypothesized to interactwith the hydroxyl group of the intermediate Mg(OH)-(OCH3) more strongly than methanol due to higher acidity, to form a materialwith a predominantly (111) surface. In the absence of the benzyl alcoholdirecting agent, (111)-oriented nanosheets were not observed. Theaddition of water induces hydrolysis and the resulting white sol\u2013gelis then transferred to an autoclave reactor where it is purged withargon and then pressurized to 10 bar before heating to 265 \u00b0C.Upon heating, the pressure in the reactor increases to reach a pseudo-supercriticalstate where it is maintained. Pseudo-supercritical drying is performedby releasing the pressure while still hot, resulting in the dry whitepowder precursor, Mg(OH)x(OCH3)x2\u2013. Calcination in air at 500\u00b0C removes all carbon species, and hydroxyl-terminated MgO(111)nanosheets are obtained.The wet chemical synthesisof MgO(111) was first reported by theRichards group via a modified aerogel method.\u03b1 radiation (\u03bb = 154 pm). Specificsurface areas were determined from nitrogen sorption isotherms acquiredat 77 K (Micromeritics ASAP 2020). Diffractograms were recorded intime intervals of 9 min. Scanning electron microscopy (SEM) measurementswere performed on a Zeiss Gemini Ultra 55 microscope operating at20 kV. The transmission electron microscopy (TEM) investigations wereperformed on a Phillips CM300 UT operated at 300 kV for all samplesexcept the MgO (111) pristine and annealed samples, which were imagedwith a cold field emission gun JEOL JEM-F200 TEM at 200 kV. The electrondose received by the samples during the electron beam irradiationexperiments was estimated from in situ measurement of the electrondose on the fluorescent screen. The TEM samples were prepared by castingsmall amounts of the dried metal oxide powders on the carbon grid.X-raydiffraction (XRD)measurements were performed on a Bruker AXS D8 Advance diffractometerusing Cu K43 and the dissociative adsorption of water results in surface energychanges that can trigger the formation of MgO nanocube stacking. Indeed,water vapor exposure of a MgO nanocube powder sample that was previouslyoutgassed at T = 1173 K and at p < 10\u20135 mbar = 30 mbar), the samples contain a large number of elongated structureswith widths that are typically larger than the size of the MgO nanoparticlesprior to H2O contact. The image in The interaction between MgO particlesand water is manifold andleads to different structures depending on the concentration of waterand its form of admission to the precleaned particle surfaces .At partial pressures in the 1\u201330 mbar range, which is comparableto those in air, water adsorption leads to coverages of a few layers,0\u20135 mbar 2a has a 2, which emerges upon water adsorption and surfacehydroxylation, as evidenced by XRD.44 XRDpattern analysis and Rietveld refinement of the data obtained on thesamples presented in 2 .The original MgO building blocks that are present in these aggregatesare covered with a characteristic shell of amorphous and crystallineMg(OH)18 Insteadof stacking into straight bars, which would be the most effectiveway to reduce their surface area, we observed regularly displacedstacks of MgO particles, appearing as staggered particle ensembles.18 Previous DFT calculations pointed to the adsorptionof water in different stages and analyzed the impact of water adsorptionon the total energy of the MgO stack: (1) Water dissociation leadsto the decoration of corners, edges, and ledges with surface hydroxyls;(2) water adsorption at terraces enables the formation of hydrogen-bondednetworks that connect to the edges and corners; (3) further wateradsorption leads to the coverage of remaining surfaces and to multilayerformation. The energetic stabilization of the displaced stacks overthe straight bars arises from water adsorption at low-coordinatedsites in combination with the decoration with MgO nanocube ledgesand residual available terrace sites.18 As a result, the staggered MgO nanocube bar, which forms duringthe early stages of hydration, serves later as a structural backbonefor the progressive conversion of the oxide into the hydroxide.In these experiments, water vapor serves as anultrapure hydroxylationagent for particle powders with precleaned surfaces. During the earlystages of their hydroxylation and hydration, MgO nanocubes self-assembleinto one-dimensional structures.45 upon contact with condensed water and whichpartly originates from the larger specific volume of Mg(OH)2. Vacuum annealing at base pressures of p < 10\u20135 mbar and T = 1173 K reconverts allhydroxides into oxides as evidenced by XRD.18 Previous FT-IR experiments performed under comparable experimentalconditions and on identical materials revealed completed dehydroxylationof the oxide surfaces. Elimination of the thermally most stable freeOH groups with their characteristic absorptions bands at \u1e7d > 3700 cm\u20131 at T >1100K and p < 10\u20135 mbar providesstrong evidence for the generation of adsorbate-free particle surfaces.47 We expect that similar decomposition,48 dehydration, and dehydroxylation processes42 are induced by electron-beam heating insidethe vacuum chamber of the TEM instrument (see the experiments on MgO(111)nanosheets described below). As a result, vacuum-annealed MgO nanocubespreviously exposed to water vapor contain characteristic elongatedbar-like structures 2 recrystallization give rise to ultrathin sheetswith high specific surface area ace area 3c,d. The2 lamella that coexist with particles of needle-likehabit that are attributed to scrolled-up Mg(OH)2 sheets.6These are a few nanometers thickand, based on the TEM data 4, can be2 has occurred in liquid water and the c axis (x110) were determined using the Scherrer equation: x001 = 3 nm and x110 = 20 nm, respectively, which are in reasonable agreement with thesheet-like morphology of the product structures observed by TEM. These,in turn, are well-suited precursor structures for the topotactic decompositionof the hydroxide into MgO.48We acquired XRD pattern on the samples, beforeand after immersioninto condensed water (id water 5b. Usingp < 10\u20135 mbar) at temperatures as low as 473K reconverts the hydroxide intothe oxide extended plate-like grainsthat partially retained the shape of the parent hydroxide flakes the values of the average crystallite domain size and (ii) the highspecific surface areas measured by sorption analysis elongated structures with lengths up to400 nm. These are attributed to partially hydroxylated stacks of staggeredMgO cubes.e flakes 6c,d. Theanalysis 1.2, which was obtained byMgO dissolution in liquid water, into MgO corresponds to a topotacticMg(OH)2 \u2192 MgO fragmentation process of the parallelhydroxide lamella into parallel polycrystalline metal oxide plates dynamichigh resolution transmission electron microscopy (D-HR-TEM) study52 that revealed atomic level details of such hydroxidedecomposition reactions and characterized the lamellar nucleationand growth processes that generate host layer bending and local elasticstrain. The resulting strain induces cracking and delamination atthe nanometer level to generate these characteristic surface topologiesof interpenetrated cubes.Thetransformation of Mg(OH)e plates6a.9,48\u221252 crystals and exhibitfaces that are oriented along the (111)planes. For samples that were treated at T = 473K in vacuum, these faces consist of regular aggregates of interpenetratedcubelets with edges in the 2\u20135 nm range, as shown via TEM via TEM 6b. The c= 1173 K 6f.SBET values of 236 \u00b1 23 m2\u00b7g\u20131 are measured surfaces reported by previous groups are not bare polar (111) surfaces but are actually stabilizedwith hydroxyl groups.55Recent publicationsin materials chemistry report the enhanced reactivity and adsorptioncapacity of polar metal oxide (111) surfaces 7a.19,20,2 nanostructures at 473 K or 1173 K produces microfaceted surfaceswith a high abundance of exposed (100) planes surface ligand during the various sample treatments.Additionally, we provide evidence that vacuum annealing of Mg(OH)) planes 7b, most 2\u00b7g\u20131 determined by nitrogen physisorption analysis. HRTEM studies providedevidence of the (111) MgO surfaces by looking at nanosheets that wereparallel to the optic axis of the TEM. Theoretical and experimentalresults strongly suggest that the polar surface is stabilized by surfacehydroxyl groups, which lowers the energy of the unstable (111) outersurface.14 In addition, previous work hasfound the surface to contain highly active corner sites, step terraces,and point defects exposing O2\u2013 anions.47 Temperature-programmed desorption (TPD) analyseswith CO2 show that the MgO(111) surface possesses primarilymedium basic Mg2+ and O2\u2013 pairs, followedby surface hydroxyl groups, while both commercial and nanoscale (\u223c4nm cubes) samples with the MgO(100) surface have weaker Lewis basesites.47To directly probe the stability of the (111) facet, we carriedout experiments to remove the stabilizing hydroxide groups from highlycharacterized MgO(111) nanosheets. The MgO(111) nanosheets probedherein are 200\u2013500 nm in diameter and 3\u20135 nm thick witha BET surface area of 200(\u00b110) m2 (more details in the 58To investigate the stabilityof bare MgO(111) surfaces, we studiedthe morphological changes of MgO(111) nanosheets described above undervacuum annealing 8 and und57 These results taken in conjunction with those above detailing thegeneration of (111) faceting via topotactic reconstruction of (100)nanocubes emphasizes that interconversion between 100 and 111 facetingis easily controllable, and thus a facile method to tune the activityfor specific applications.We made a similar observation under sustained electron-beam irradiationduring TEM investigation of the pristine MgO(111)sample, as seen by the progressive transformation of the MgO(111)surfaces into MgO(100) cubes during imaging 9. This c2O adsorption-induced organization of MgOnanocubes and their further dissolution\u2013recrystallization intowell-defined nanostructures represent an attractive approach to generateextended faceted structures of cubic ionic metal oxide nanoparticleswith rock salt structure.H2 sheet formation and sheet exfoliationin pure water are natural and generic processes that do not requirespecific counterions or surfactant species.Mg(OH)2 nanosheets can easily undergo decompositionand faceting to transform into MgO grains and platelets with uniformlyshaped grain faces that can be described as inverse cube elements.This topotactic conversion of hydroxide into the metal oxide can beachieved on products of different synthesis approaches and Mg/MgOprecursors, such as vapor-phase grown MgO cubes or sol\u2013gel-derivedMgO(111) nanosheets (Mg(OH)nosheets 6 and 8.Wateradsorption at the solid\u2013gas interface of MgO nanostructuresand their subsequent dissolution in gaseous or liquid water can produce1D and 2D structures, and particles of high morphological definition,with staggered and interpenetrated nanocubes as primary building blocks.This study reports on the different steps involved in these transformationsand demonstrates the following:59The availability of MgO(111) nanosheets as well as MgO-based nanocubearrays, where the cubic building blocks are organized at differentlevels of order, ranging from randomly oriented particles within drypowders, 1D bars of staggered nanocubes, to reconstructed facetedsurfaces of inverse cubes, opens a range of opportunities for adsorptionstudies and heterogeneous catalysis research.2O molecules from the gas phase through pumping during annealingeffectivelysuppresses particle growth and coarsening. Thus, thermally stableporous microstructures exclusively composed of MgO nanocubes can besynthesized with a variety of configurations without compromisingthe parent particle size. In addition to the potential of such architecturesas substrates for heterogeneous catalysis, our findings are relevantfor the development of sintering approaches to manufacture and functionalizenanocrystalline ceramics.Furthermore, we show that the continuous desorption of H8 planesand thus, a high abundance of edge features with four-coordinatedions, should also encompass cooperative effects between adsorbatesand the different surface elements. Moreover, calculations to assessthe energetics of the topotactic transformation process (rather thanthe octopolar reconstruction16) in relationto the energy required for the decomposition of the most stable surfacehydroxyls would be most useful.Future theoretical and experimentalstudies on the reactivity ofthe inverse cube arrays featuring MgO56 requires further attention.Finally, our results show thatMgO(111) nanostructures reconstructinto MgO(100) surfaces upon surface adsorbate removal, suggestingthat these MgO(111) surfaces are not bare and thus not polar undersurface science experimental conditions. We believe that the importantquestion of how charge-compensating surface groups such as hydroxylsor methoxy groups can enhance and promote the catalytic activity andadsorption capacity as reported for MgO(111) nanosheets"} +{"text": "The im phenotype is caused by a single recessive mutation of a pentatricopeptide repeat (PPR) gene that reduces the activity of mitochondrial complex I and up-regulates stress responsive genes. However, the mechanisms altering the stress responses in im mutant are not well understood. Thus, we characterized growth and gas exchange in im and TM-1 under no stress and also investigated their stress responses by comparing gas exchange and transcriptomic profiles under high temperature. Phenotypic differences were detected between the NILs in non-fiber tissues although less pronounced than the variation in fibers. At near optimum temperature (28\u00b13\u00b0C), im maintained the same photosynthetic performance as TM-1 by means of greater stomatal conductance. In contrast, under high temperature stress (>34\u00b0C), im leaves reduced photosynthesis by decreasing the stomatal conductance disproportionately more than TM-1. Transcriptomic analyses showed that the genes involved in heat stress responses were differentially expressed between the NIL leaves. These results indicate that the im mutant previously reported to have low activity of mitochondrial complex I displays increased thermosensitivity by impacting stomatal conductance. They also support a notion that mitochondrial complex I activity is required for maintenance of optimal photosynthetic performance and acclimation of plants to high temperature stress. These findings may be useful in the future efforts to understand how physiological mechanisms play a role in determining cotton fiber maturity and may influence stress responses in other crops.Thickness of cotton fiber, referred to as fiber maturity, is a key determinant of fiber quality, lint yield, and textile performance. The cotton immature fiber ( Gossypium sp.) is the world\u2019s most important natural fiber with (A) stomatal conductance [gs] and (B) net photosynthesis [Pnet] from the field-grown im and TM-1 leaves.(PDF)Click here for additional data file.S1 TableArabidopsis sequences (TAIR 10) and annotated based on the functions of the Arabidopsis genes that were the best hit by BLAST search.The sequences of the cotton DEGs were compared with (XLSX)Click here for additional data file.S2 TableG. hirsutum mitochondrial genomes (JX065074).Cotton mitochondria genes were compared with the (XLSX)Click here for additional data file.S3 TableG. hirsutum plastid genomes (DQ345959).Cotton chloroplast genes were compared with (XLSX)Click here for additional data file.S4 TableArabidopsis sequences (TAIR 10) and annotated based on the functions of the Arabidopsis genes that were the best hit by BLAST search.The sequences of the cotton DEGs were compared with (XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S1 Graphic abstract(TIF)Click here for additional data file."} +{"text": "An ongoing prospective phase 2 trial (NCT01954173) is examining the role of postoperative photon radiation therapy for high-risk patients using volumetric modulated arc therapy. Proton beam therapy (PBT) may be beneficial in this setting to reduce hematologic toxicity. We evaluated for dosimetric relationships with pelvic bone marrow (PBM) and changes in hematologic counts before and after pelvic radiation therapy and explored the potential of PBT treatment plans to achieve reductions in PBM dose.For patients with high-risk bladder cancer (pT3t test. Correlation of mean nadirs and relative PBM dose levels were assessed using the Pearson correlation coefficient (CC).All enrolled patients were retrospectively analyzed after pelvic radiation per protocol with 50.4 to 55.8 Gy in 28 to 31 fractions. Comparative PBT plans were generated using pencil-beam scanning and a 3-beam multifield optimization technique. Changes in hematologic nadirs were assessed using paired P\u2009<\u2009.001), and platelets (P\u2009=\u2009.03). Increased mean PBM dose was associated with lower nadirs in WBC , ANC , and hemoglobin , whereas the PBM V30 to V40 correlated with lower WBC , and V20 to V30 correlated with lower ANC . Comparative proton therapy plans decreased the mean PBM dose from 26.5 Gy to 16.1 Gy (P\u2009<\u2009.001) and had significant reductions in the volume of PBM receiving doses from 5 to 40 Gy (P\u2009<\u2009.001).Eighteen patients with a median age of 70 were analyzed. Mean cell count values after radiation therapy decreased compared with preradiation therapy values for white blood cells (WBCs), absolute neutrophil count (ANC), absolute lymphocyte count (all Increased PBM mean dose and V20 to V40 were associated with lower hematologic nadirs. PBT plans reduced PBM dose and may be a valuable strategy to reduce the risk of hematologic toxicity in these patients. Although having the advantage of evaluating patient data prospectively collected during a clinical trial, this study included a relatively small patient cohort and was unable to report physician- or patient-reported clinical toxicity to corroborate the clinical implications of these cell nadirs. These results will be reported in future primary analysis of NCT01954173 . In addie target . Ultimate target closed ee target and the e target are ongoSecondary analysis of the ongoing phase 2 clinical trial NCT01954173 demonstr"} +{"text": "Circovirus genus, the first reported in genus Equus. Whether EqCV1 alone or in co-infections can result in disease and its prevalence in different equine populations will require further studies now facilitated using EqCV1\u2032s genome sequence.Circoviruses infect vertebrates where they can result in a wide range of disease signs or in asymptomatic infections. Using viral metagenomics we analyzed a pool of five sera from four healthy and one sick horse. Sequences from parvovirus-H, equus anellovirus, and distantly related to mammalian circoviruses were recognized. PCR identified the circovirus reads as originating from a pregnant mare with fever and hepatitis. That horse\u2019s serum was also positive by real time PCR for equine parvovirus H and negative for the flavivirus equine hepacivirus. The complete circular genome of equine circovirus 1 strain Charaf (EqCV1-Charaf) was completed using PCR and Sanger sequencing. EqCV1 replicase showed 73\u201374% identity to those of their closest relatives, pig circoviruses 1/2, and elk circovirus. The closest capsid proteins were from the same ungulate circoviruses with 62\u201363% identity. The overall nucleotide identity of 72% to its closest relative indicates that EqCV1 is a new species in the Circoviridae family whose circular single-stranded DNA genomes of ~2 kb are amongst the smallest known [Circoviruses are small viruses in the st known . Circovist known ,18,19,20st known ,26,27,28Here we report on a new circovirus we called equine circovirus 1 strain Charaf (EqCV1-Charaf) identified during a viral metagenomics analysis of plasma from horses.Viral-like particles were enriched by filtration and digestion with nucleases enzymes to reduce the concentration of non-capsid protected nucleic acids. Following nucleic acids extraction and random RT-PCR the DNA amplification products were converted to Illumina compatible DNA using Nextera\u2122 XT Sample Preparation Kit with dual barcoding and analA pair of PCR primers was designed based on circovirus-related reads to generate a 352 bp amplicon and used to test nucleic acids extracted individually from each of the five sera in the pool. The DNA genome of the circovirus was then completed using two PCR over gaps with primers designed from the short reads (available on request) and the amplicons directly Sanger sequenced by primer walking. Sequence analysis was performed using MAFFT multiple sequence alignment . Sequenc\u221227 indicating the detection of a \u201cnovel\u201d circovirus. Sera from five horses from Kentucky were pooled and analyzed by viral metagenomics. Four horses were clinically healthy while one had fever and hepatitis. Analysis of the resulting 120,000 Illumina sequence reads showed the presence of fifteen reads to anellovirus Torque teno equus virus 1, two reads to the recently described equine parvovirus H and forty-five reads with best hits to different circoviruses. Because anelloviruses are considered commensal viruses and the genome and pathogenicity of equine parvovirus H are well characterized we focused on the circovirus hit with BLASTx E score ranging from 0.003 to 10Each plasma sample was individually extracted and tested by PCR for the presence of circovirus DNA. Only one serum collected from a 10 months pregnant mare stabled in central Kentucky was PCR positive. The circovirus infected horse exhibited pyrexia and inappetence with elevated liver enzymes GGT (Gamma Glutamyl Transferase) and SDH (Sorbitol dehydrogenase) activities. The horse responded favorably to NSAIDS with Flunixin PRN for fevers, antibiotics TMS tablets (Trimethoprim 160 mg and Sulfamethoxazole 800 mg) 24 mg/kg BID, Pentoxifylline 4 g BID, Vitamin E for at least 3 weeks. Real time PCR tests performed by Cornell University Animal Health Diagnostic Center reported negative results for the flavivirus equine hepacivirus and positive results for equine parvovirus (EqPV-H) with Ct values of 32.36. EqPV-H, a frequent equine infection ,35, has PCRs followed by amplicons Sanger sequencing were then used to fill two gaps in the genome allowing the complete equine circovirus (EqCV) genome to be assembled. The EqCV1 genome was 1788 bases long, with a GC content of 53%, with three major ORFs encoding replicase (Rep) capsid (Cap) proteins and ORF3 of unknown function (GenBank MW881235) . The Rep protein was 327 amino acids long, sharing aa-identities of 74% with elk circovirus (Genbank MN585201) followed by PCV2 (GenBank EU148505.1) with 73% identity. The 231 amino acid long capsid protein was also closest to elk circovirus capsid (63%) (GenBank MN585201.1) followed by PCV1 (GenBank NP_065679.1) with 61.7%, PCV2 (GenBank NP_937957) (60.2%), and bat associated circovirus 2 (GenBank YP_007974238) with 48% identity. A third major ORF of a 274 amino acid protein was also identified located antisense over the rep ORF. Its closest known relative is the ORF3 from PCV2 (GenBank YP_006355434.1) at 54% identity over 38% of protein. This protein found in PCV1 (204 amino acids) and PCV2 (104 amino acids) is believed to trigger apoptotic activity in infected cells . The Rep contained all three expected rolling circle replication motifs I (VFTLNN), II (PHLQG) and III (YCGK) . The C-tPhylogenetic analyses were then performed showing that the EqCV1 rep and cap proteins clustered with PCV1/2, as well as elk and bat 2 (insectivorous Rhinolophus ferrumequinum or greater horseshoe bat) circoviruses . According to the ICTV, the member of the same circovirus species should share >80% nucleotide identity over their entire genome, and >70% amino acid identity between their Cap proteins . When usThe equine circovirus genome characterized here is most closely related to a Canadian elk circovirus reported once and to pig circoviruses PCV1 and PCV2 distributed world-wide. Four porcine circovirus species are currently known that are either non-pathogenic (PCV1), pathogenic (PCV2), possibly pathogenic (PCV3) ,43, or sThe common detection of PCV1 and PCV2 in pig world-wide thereforAttributing disease causality to a new virus without fulfilling Koch\u2019s postulates is challenging. Typically, this begins with elimination of pathogens known to be able to cause the observed lesion, and is followed by epidemiologic, clinical, pathological and microbiological investigations that test this new virus-disease association. Here a horse with a mild case of hepatitis was infected with a circovirus but also viremic at low levels with the hepatotropic EqPV-H. Although many asymptomatic EqPV-H infections have been reported EqPV-H d"} +{"text": "Nitrogen containing heterocyclic compounds has acquired their remarkable and distinct place in the wide area of organic synthesis due to the broad range of applications. Among them, quinoline motifs have attracted researchers in the synthetic chemistry because of its presence in the large number of pharmacologically active compounds. Different methods for synthesis of quinoline derivatives are reported, among them the Friedlander synthesis have provided comparatively more efficient approach. Many of the reported conventional Friedlander methodologies have some problems such as difficult product isolation procedures, poor yields and use of expensive catalysts, etc. Recently, polymer or solid supported synthetic approaches have attracted the attention of researchers because of their easy execution, greater selectivity, increased product yields, simple work-up procedures, recoverability and reusability of the catalysts. In consideration with the advantages of polymer supported synthetic strategies, the proposed review covers the role of polymers in the Friedlander synthesis; which may use polymers of organic, inorganic or hybrid in nature and of nanolevel as well. Quinoline scaffolds occur in large number of pharmacologically active compounds obtained from natural sources as well as prepared synthetically. The compounds containing quinoline moiety like pamaquine, chloroquine, tafenoquine, bulaquine, mefloquine, piperaquine, pyronaridine are the potent antimalarial agents and amodiaquine as an antimalarial and antiinflammatory agent. The quinoline derivatives ciprofloxacin and levofloxacin marketed as broad-spectrum antibiotics [1]. Quinoline derivatives has been reported to possess diverse variety of medicinal activities such as anticancer , antibacterial [4], DNA binding [5], antiproliferative [6], antitumor , antitubercular [9], anticonvulsant, antihypertensive [10], antiinflammatory, cardiovascular [11], antimalarial, antiprotozoal [12], antifungal [13], antioxidant [14], anti-HIV agents [15] and for treatment of central nervous system diseases [16]. The varieties of drugs containing quinoline pharmacophore are shown in Figure 1.The first synthesis of quinoline was attempted using aniline as crucial reagent by Skraup (1880); align to this classical synthetic route several methods are reported as the advancement for synthesis of quinoline moieties. The synthetic methodologies which used aniline include Skraup reaction [17], Combes reaction [18], Conrad\u2013Limpach reaction [19], Povarov reaction [20], Doebner reaction [21], Doebner\u2013Miller reaction [22], Gould\u2013Jacobs reaction [23] and Riehm reaction [24] (Scheme 1).Several synthetic routes were reported which involved modification to aniline based protocols. The substituted anilines or some typical substituted reactants are used for synthesis of quinoline motifs which includes Knorr reaction [25], Pfitzinger reaction [26], Friedlander reaction [27], Niementowski reaction [28], Meth\u2013Cohn reaction [29] and Camps reaction [30] (Scheme 2).3) and its derivatives. The classical Friedlander reaction reported by Paul Friedlander in 1882, used 2-aminobenzaldehyde (1) with acetaldehyde (2) in the presence of sodium hydroxide [31] (Scheme 3). Generally, Friedlander reaction is carried out by refluxing aqueous or alcoholic reaction mixture in the presence of a base or an acid or by heating reaction mixture at higher temperature without catalyst [32]. Lewis acids were also used commonly and found efficient . The Friedlander synthesis is one of the simplest methods among the several reported methods for the synthesis of quinoline and ketones (11). The Boc-protected aminophenol (5) was treated with commercial TentaGel-Br resin (4) to afford resin 6, which on cleavage of Boc with TFA in DCM furnished the resin 7. The supported imine 10 was prepared in two steps from resin 7 by treatment with 3,4-dimethoxy-6-nitrobenzaldehyde (8) in refluxing ethanol, affording resin-bound o-nitroimine (9) and subsequently on reduction of the nitro group in the presence of sodium sulfide in refluxing ethanol to obtain the desired resin-bond \u201cmasked\u201d o-benzaldeimine (10). The resin bound imine (10) then treated with different ketones in the presence of piperidine under refluxing ethanol which underwent cyclative cleavage to yield substituted quinolines (12) (Scheme 4). Average to good yields of corresponding quinolines was obtained.Patteux et al. (2003) [39] described the solid-phase synthesis of quinolines via a modified Friedlander reaction between the resin-bound imine (13) with 2-aminoarylketones (14) in the presence of polyphosphoric acid (PPA) as a catalyst [40]. The PEG bound quinoline-3-carboxylates (15) were obtained from the reaction of 13 with 14, which subsequently underwent cleavage from the PEG in the presence of MeONa in MeOH to afford polysubstituted quinolines 16 (Scheme 5). Excellent yields of products were reported. Zhang et al. (2011) described microwave assisted synthesis of polysubstituted quinolines using polyethylene glycol (PEG 4000) bound acetoacetate under refluxing conditions to afford quinolines 20 by a Friedlander-type reaction with cyclative cleavage of solid support (Scheme 6). Yamaguchi et al. (2015) reported solid phase Friedlander synthesis using an alkoxyamine linker [41]. The 2-aminobenzoyl compounds N-bromo-N-ethylbenzene-1,3-disulfonamide) [PBBS] and N,N,N\u2019,N\u201d-tetrabromobenzene-1,3-disulfonamide [TBBDA] as efficient catalysts [42]. Both the aqueous and solvent free conditions affording excellent yields of products but, the aqueous protocol occurred in shorter time in comparison with to solvent free. Ghorbani-Vaghei and Akbari-Dadamahaleh (2009) achieved Friedlander synthesis under aqueous and solvent free conditions using poly(4), Figure 2B, Table 1] as an efficient dual acidic catalyst for the one-pot synthesis of quinolines under solvent free conditions via Friedlander reaction [43]. The solvent free reaction was attempted in the presence of P(4VPBSA)-HSO4 as a catalyst. In spite of high temperature conditions, the method reported short reaction time and excellent yields.Kiasat et al. (2013) employed poly(4-vinylpyridiniumbutanesulfonic acid) hydrogensulfate [P(4VPBSA)-HSOFang et al. (2013) investigated branched catalysts with hydrocarbon or fluorocarbon chains as a catalyst for Friedlander reaction under solvent free conditions [44]. The Friedlander components were heated neatly in the presence of dendritic hydrocarbon chains or fluorocarbon chains as a catalyst at 80 \u00b0C. After heterogeneous work up with ethyl acetate, good to excellent yields of corresponding substituted quinolines were obtained. Both the catalysts seem to be an equally efficient. The discussed methodology contributed new form of branched catalyst to the advancement in the Friedlander synthesis.N,N-methylene bis-acrylamide (MBA), 2-acrylamido-2-methylpropanesulfonic acid (AMPS) and acrylic acid (AA) by earlier reported method. Although the methodology involved high temperature conditions, it has been added new polymeric catalyst to the Friedlander catalyst library.Maleki et al. (2015) investigated cross-linked polymer bound sulfonic acid as a catalyst for synthesis of substituted quinolines via Friedlander reaction under solvent free conditions [45]. The catalyst Poly (AMPS-co-AA) was prepared from cross-linked 21) and ketones (22) in the presence of KOH using dioxane as solvent via oxidative cyclization [46]. Although the method seems conventional with use of solvent, longer reaction times and less economic yields, but it has described the new modified Friedlander approach with the use of Pd/C as a catalyst.Cho et al. (2005) reported Pd/C catalyzed modified Friedlander reaction using 2-aminobenzylalcohol as a catalyst for the synthesis of multisubstituted quinolines by the Friedlander heteroannulation under solvent free conditions [47]. The method highlighted the entry of new carbon supported catalyst with excellent yields of products, short reaction times and reusability of catalyst for three cycles with negligible loss of activity.Shirini et al. (2013) employed rice husk ash supported FeCl3H, Table 1) was reported as a catalyst for the Friedlander reaction reported by Shirini et al. (2015) under solvent free conditions [48]. The catalyst RHA-SO3H was prepared by the reaction between rice husk ash (RHA) and chlorosulfonic acid (ClSO3H). The recyclability of the catalyst for three consecutive cycles was mentioned. The method seems to be an efficient and provided high yields in short reaction times. The sulfonated rice husk ash resins as a catalyst for Friedlander quinoline synthesis [49]. This was a very first attempt of using ion exchange resins as a heterogeneous catalyst for the synthesis of substituted quinolines using Friedlander reaction. The Friedlander components were refluxed in ethanol in the presence of Amberlite IRA 400 or Dowex 2 resins as catalyst; good quantities of products were obtained. Das et al. (2007) reported the Friedlander synthesis of substituted quinolines using Amberlyst-15 resin as a solid heterogeneous catalyst [50]. The Friedlander reactions were performed under refluxing ethanol in the presence of the Amberlyst-15 catalyst. The reusability of catalyst for three subsequent cycles was mentioned with slight declined activity. Although the method seems conventional; it has contributed a new form of catalyst to the advancements in the Friedlander reaction.n-butyl-3-methylimidazolium hexafluorophoaphate, [Bmin] [PF6] ionic liquid as a medium for the Friedlander reaction in the presence of catalyst Amberlyst-15 [51]. The methodology seems conventional although it has introduced new combination of catalyst and ionic liquid as a medium.Hou et al. (2008) attempted ionic liquid mediated Friedlander quinoline synthesis using Amberlyst-15 resin as a catalyst. The method employed 1-Wang et al. (2011) performed solvent free Friedlander annulation with an ion exchange resin, Dowex-50W as a catalyst[52]. The heterogeneous phase work up procedure affording good amount of product yields. Shorter reaction times with solvent free condition make the method efficient, but the high temperature requirement for the catalytic efficiency makes it more common.3H, Table 1). The mixture of 2-aminoarylketone, \u03b1-methyleneketone and PEG-SO3H as a catalyst was refluxed in DCM; excellent amounts of products were obtained. The reusability of recovered catalyst was mentioned.Zhang et al. (2009) [53] reported Friedlander synthesis using poly(ethylene glycol) (PEG-4000)-supported sulfonic acid as a catalyst for the microwave irradiated Friedlander annulation under solvent free conditions [54]. Well ground mixture of Friedlander components including PEG-OSO3H was subjected to irradiation in microwave oven at 600W; excellent amounts of substituted quinolines were obtained. The recyclability of the recovered catalyst for many subsequent cycles was found. The method reported excellent yields under solvent free conditions as well as an efficient reusability of the catalyst. Hasaninejad et al. (2011) demonstrated use of sulfuric acid-modified PEG-6000 (PEG-OSO3H (Table 1) as a catalyst in aqueous medium. The aqueous mixture of 2-amino-5-chlorobenzophenone or 2-aminobenzophen- one, active methylene compounds and PEG-SO3H was stirred at 60 \u00b0C; affording good to excellent yields of substituted quinolines [55]. The recovered catalyst was reused for six subsequent cycles with slight decrease in activity after each rerun. Variety of active methylene compounds were used, both in cyclic and acyclic forms thereby contributed a rich library of quinoline scaffolds to the literature. The use of green solvent, short reaction times, economic yields, simple work up and reusability of catalyst makes the protocol remarkable for the improvements in the Friedlander reaction.Nasseri et al. (2013) demonstrated Friedlander synthesis using PEG-SO5.PEG, Table 1) and niobium (V) chloride (NbCl5) as a catalyst in glycerol as a solvent [56]. Good to excellent amounts of yields were reported. The reusability of recovered catalyst was mentioned. Both the catalyst NbCl5.PEG and NbCl5 proved noticeably efficient. Although the methodology claimed greener approach but use of solvent at higher temperature and usual work up procedure makes it align to conventional and limits the credit.Zakerinasab et al. (2015) achieved Friedlander synthesis using niobium-(V) chloride /- polyethylene glycol (NbCl25), dialkyl acetylenedicarboxylate (26) and catalyst \u03b2-CD was heated, in which excellent yields of polysubstituted quinolines (27) were obtained (Scheme 8). The reusability of recovered catalyst for four consecutive cycles with slight decrease in activity after each re-run was mentioned. The protocol employed greener approach and seems conventional with solvent extraction work up procedure.Cyclodextrins are able to form host-guest complexes with hydrophobic molecules. Different forms of attractive forces like van der Waals interactions are believed to involve in the formation of inclusion complex between guest molecule and CDs [57]. Madhav et al. (2010) explored modified Friedlander synthesis using supramolecular catalyst \u03b2-CD (Table 1) in aqueous medium [58]. The aqueous reaction mixture containing substituted 2-aminobenzophenone as a heterogeneous catalyst for the synthesis of 1,2,3-trisubstituted quinolines via solvent free Friedlander annulation [59]. Excellent amounts of yields were reported. The efficient reusability of recovered catalyst for three successive runs was reported. New catalyst system has been added to the advancements in the Friedlander reaction although the method involved longer reaction times.Desai et al. (2006) applied silica gel supported sodium hydrogen sulphate employed silica sulfuric acid as a solid acid catalyst under solvent free conditions for Friedlander reaction [60]. Simple work up, economic yields, solvent free condition, short reaction times and reusability of SSA catalyst highlighted the valuable improvement in the Friedlander reaction.p-toluenesulfonic acid supported on silica gel as a heterogeneous catalyst [61]. The anthranilonitrile (28) and cyclohexanone (29) was reacted in the presence of the catalyst p-TSA/SiO2 under solvent free conditions; affording good yields of substituted 9-amino-1,2,3,4- tetrahydro-acridines (30) (Scheme 9). Variety of solid supports were screened to optimize of the catalytic activity such as acidic alumina, zeolite (HY), montmorillonite (K-10) and silica gel; among them silica found most suitable. The p-TSA/SiO2 catalyst system was evaluated under both conventional and microwave conditions; microwave irradiated protocol was found to be comparatively more efficient.Khalilzadeh et al. (2007) attempted microwave assisted solvent free Friedlander quinoline synthesis using 4-SiO2, Table 2) as catalyst for the synthesis of quinoline derivatives by Friedlander heteroannulation [62]. Acetonitrile was used as solvent, and the method provided excellent yields. Recovered catalyst reused for three cycles. Albeit, the discussed HClO4-SiO2 catalyst approach seems conventional, it has contributed new efficient catalyst to the advancements in the Friedlander synthesis. Narasimhulu et al. (2007) investigated silica supported perchloric acid as a heterogeneous catalyst for the solvent free Friedlander heteroannulation [63]. The methodology was found to be added a new form of silica supported catalyst but seems slightly less impressive due to average to good yields and lack of reusability of catalyst but the solvent free reaction at low temperature conditions marks it entry to the catalyst library for the advancement of Friedlander reaction. Zolfigol et al. (2007) evaluated molecular iodine supported on silica gel under solvent free conditions at higher temperature compared to earlier reported method [64]. Excellent yields of substituted quinolines were obtained in short reaction times. The efficiency recovered catalyst was retained for three consecutive cycles.Dabiri et al. (2007) described improved Friedlander synthesis using sodium hydrogen sulphate as catalyst [65]. The ethanolic reaction mixture containing Friedlander components was refluxed in the presence of PMA-SiO2 catalyst; affording good amounts of products. The reusability of catalyst was mentioned without frequency and efficiency. Das et al. (2008) attempted conventional Friedlander synthesis using silica supported phosphomolybdic acid (PMA-SiO4)2\u00b712H2O-SiO2 as a catalyst [66]. Variety of active methylene compounds were used for the synthesis; furnished excellent amounts of corresponding quinolines. The recovered catalyst was recycled for six subsequent runs with negligible loss of activity after each run. The Alum-SiO2 catalyzed protocol seems to be attractive due to solvent free and low temperature conditions along with excellent yields and reusability of the catalyst.Mohammadi et al. (2008) reported solvent free synthesis of polysubstituted quinolines using KAl demonstrated microwave assisted solvent free Friedlander synthesis using silica sulfuric acid as a heterogeneous catalyst [67]. The neat Friedlander reaction mixture including SSA as a catalyst was microwave irradiated (900W); furnished good to excellent amounts of substituted quinolines. The solvent free condition and shorter reaction times makes the method remarkable. 2O5 as a catalyst for solvent-free synthesis of poly-substituted quinolines via Friedlander heteroannulation reaction [68]. Good to high amount of quinolines were reported. Solvent free condition and short reaction times makes the method attractive.Hasaninejad et al. (2011) reported silica-supported P2, Table 2) as catalyst [69]. The mixture of 2-aminoacetophenone (31), one drop of diluted hydrochloric acid and silica gel was subjected to microwave irradiation at 440 watts for 0.5 h; affording dimerization product 32 (Scheme 10). The same methodology was employed for the reaction between 31 and substituted 2-hydroxyacetophenone; average yields of corresponding quinolines were reported.Al-Qahtani et al. (2013) proposed microwave assisted synthesis of polysubstituted quinolines by Friedlander reaction using dilute hydrochloric acid and silica gel for the preparation of polyfunctional quinoline derivatives under solvent free conditions and ultrasonication [70]. The Friedlander reaction components added with SiO2-Cl catalyst was sonicated neatly in a SIDILU sonicator (35 KHz and 120 W); furnished good yields of substituted quinolines. The reusability of catalyst was reported for two subsequent cycles with continuous decrease in activity. The protocol added new form of silica-based catalyst to the advancement in the Friedlander reaction. In spite of less economic yields the method seems attractive due to use of solvent free and ultrasonication conditions. Sudha et al. (2013) investigated the use of silica chloride attempted microwave assisted solvent free synthesis of polycyclic quinoline derivatives using montmorillonite KSF clay (Table 2) as a catalyst [71]. The solvent free reaction mixture containing Friedlander substrates and KSF clay catalyst was microwave irradiated (600W); affording considerable amounts of corresponding quinolines. Typical active methylene compounds were used to prepare polycyclic quinoline derivatives. In spite of less economic yields; the solvent free condition, short reaction times and addition of new quinoline scaffolds to the quinoline library makes the methodology creditable.3.nH2O, hydrotalcite (HT), Mg6Al2(OH)16CO3 and triethylamine. The variety of catalyst systems were screened for optimization of the reaction such as HT, Ru/Al2O3, Ru/MgO, Ru/ Mg(OH)2, and Ru/Al(OH)3 among them, Ru/HT-N (Table 2) proved to be most efficient. The reaction mixture of 2-aminobenzyl alcohol (21), active methylene compounds (11) and catalyst Ru/HT-N was vigorously stirred at 100 \u00b0C in toluene under an O2 atmosphere for 20 h; affording good amounts of corresponding quinolines (34) (Scheme 11). Reusability of recovered catalyst was mentioned with consistent catalytic activity. Although the protocol seems conventional and less impressive due use of toxic solvent and longer reaction times, it has contributed another modified approach aligns to the Friedlander heteroannulation. Motokura et al. (2004) demonstrated one pot two step synthesis of quinolines using Ru-grafted hydrotalcite as a catalyst via modified Friedlander reaction [72]. The catalyst Ru/HT was prepared in the presence of triethylamine and labelled Ru/HT-N using RuCl.L\u00f3pez-Sanz et al. (2010) reported zeolites (HY) as a catalyst for Friedlander quinoline synthesis. The zeolites H-BEA, H-MFI, H-FAU and H-MOR were investigated for catalytic performance; among them H-BEA and H-FAU (Table 2) were found to be the most efficient catalysts for the quinoline synthesis [73]. The yields were average to good. Under both the conditions (using toluene as solvent and solvent free), the Friedlander reactions were performed in the presence of zeolite catalyst; the solvent free protocol found more efficient3H, Table 2) as an aluminosilicate mineral based catalyst under solvent free conditions [74]. Excellent amounts of quinoline derivatives were obtained in short reaction times. The reusability of catalyst MCM-41-SO3H was mentioned for several consecutive cycles without considerable loss in activity. The protocol found to be added an efficient catalyst to the advancement in the Friedlander reaction.Maleki et al. (2014) proposed Friedlander annulation using silica based sulfonic acid assisted protocol used MK-10 catalyst under solvent free conditions. The later solvent free protocol proved to be most efficient compare to conventional approach by showing excellent yields in shorter reaction times.Subashini et al. (2014) reported microwave assisted solvent free Friedlander synthesis of quinolinyl quinolinones by using montmorillonite K-10 as a heterogeneous catalyst [75] (Scheme 12). The synthesis of quinolinyl quinolinones was performed under conventional as well as microwave irradiated conditions. The conventional synthesis employed use of conc. H3PW12O40, Table 2) as a heteropoly acid-based catalyst for the Friedlander quinoline synthesis [76]. The ethanolic reaction mixture containing Friedlander components was refluxed in the presence of Ag3PW12O40 catalyst; affording good amount of yields of substituted quinolines. Varieties of active methylene compounds were used. The recovered catalyst was recycled after activation for four consecutive cycles with gradual decrease in activity after each rerun. Although the protocol seems conventional with use of solvent and longer reaction times; it has successfully added the new form of catalyst system to the improvement in the Friedlander reaction. Yadav et al. (2004) utilized silver phosphotungstic acid as a heterogeneous catalyst [77]. Different active methylene compounds were used to prepare 2,3,4-trisubstituted quinolines. Excellent amounts of desired quinolines were reported. The reusability of the recovered catalyst was mentioned for five subsequent runs with almost consistent catalytic activity. The solvent free condition, efficient reusability of catalyst and excellent yields has highlighted the noticeable advantages of the discussed method.Dabiri et al. (2009) attempted solvent free Friedlander synthesis using phosphotungstic acid (H6 [P2W18O62] (Table 2) as a solid acid catalyst for the synthesis of quinolines via Friedlander reaction under solvent free conditions [78]. The catalytic activity of heteropolyacids such a H6[P2W18O62], H5[PMo10V2O40] and H6[PMo9V3O40] were screened for catalytic performance; H6 [P2W18O62] proved most efficient with excellent yields. The reusability of catalyst for five consecutive cycles was mentioned with gradual decrease in activity after each cycle. Heravi et al. (2010) evaluated role of Wells\u2013Dawson type of heteropoly acid H2.5H0.5PW12O40, Table 2), a heteropoly salt as a catalyst for the Friedlander reaction under solvent free conditions [79]. The acidic salts of CsxH3-xPW12O40 were prepared using literature method; among all the Cs2.5H0.5PW12O40 salt found to be more efficient catalyst. The solvent free condition, excellent yields, short reaction times and reusability of the catalyst make the method advantageous over conventional approaches. The same group has reported Keggin type heteropoly acids (HPAs) and supported ones on solids with different nature and textural properties as a catalyst under solvent free conditions for Friedlander synthesis [80]. The tungstophosphoric acid supported on silica (PW/SiO2), KSF (PW/KSF), activated carbon (PW/C) and \u03b3-alumina (PW/\u03b3-Al2O3) (Table 2); among these all the catalyst system found to be equally efficient except PW/\u03b3-Al2O3. Reusability of all the catalyst systems was reported for four consecutive cycles with gradual decrease in activity after each cycle. The article contributed efficient catalyst systems to the advancement of Friedlander reaction. The solvent free condition, excellent yields, shorter reaction times, reusability of catalyst seems creditable.Rafiee et al. (2011) employed cesium partially substituted phosphotungstate as a catalyst for the Friedlander synthesis of novel heteroatom substituted quinolines under solvent free conditions [81]. The substituted 2-aminoaryl ketones (14) and a heteroatom bearing ketones (38) was heated solvent free in the presence of H3PW12O40 catalyst; affording corresponding quinoline scaffolds (39) (Scheme 13). The recovered catalyst was reused for five subsequent cycles with slight decrease in activity up to third cycle and thereafter it decreases considerably. Although the products were average to good, the authors have reported a rich library of heteroatom substituted quinoline scaffolds which make the valuable contribution.Chen et al. (2014) investigated use of phosphotungstic acid (H40) were treated with different anhydrides (41) PPA; affording corresponding quinolines (42) (Scheme 14). Though yields of quinolines were average to good, but the protocol has proposed new route for synthesis of quinolines with a modified Friedlander approach.Na et al. (2005) demonstrated modified Friedlander synthesis of quinolines in the presence of PPA (Table 2) [82]. The substituted enaminones (14), substituted 3-cyanoacetylindoles (43) and catalyst PPA in PEG-400 was subjected to thermal or microwave irradiation (350W); furnished good yields of corresponding 2-(indol-3-yl)-3-nitriloquinolines (44) (Scheme 15). Compare to thermal method, microwave protocol found to be most efficient with shorter reaction times and higher yields. The use of green solvent and contribution of diverse quinoline scaffolds to the library highlighted the valuable advancement to the Friedlander annulation.Shiri et al. (2012) reported synthesis 2-(indol-3-yl)-3-nitriloquinolines using PPA (Table 2) as a catalyst via modified Friedlander heteroannulation under thermal as well as microwave assisted conditions [83]. The mixture of substituted amino ketones as a catalyst [84]. The mixture of 2-aminonicotinaldehyde (45), active methylene compound (46) and basic Al2O3 was ground by pestle and mortar at room temperature; affording excellent yields of 1, 8-naphthyridines (47) (Scheme 16). Short reaction times, excellent yields and use of mortar-pestle provide the creditable improvement in the Friedlander reaction.Mogilaiah and Vidya (2007) proposed Friedlander synthesis of substituted 1, 8-naphthyridines in the solid state using basic alumina demonstrated Friedlander quinoline synthesis using basic mesoporous amino-grafted MCM-41 as hybrid catalysts [85]. The variety of MCM-41 materials supporting aminopropyl (AP), methylaminopropyl (MAP), and diethylaminopropyl (DEAP) groups were prepared and also modified the acidic properties of Al-SBA-15 by incorporating cesium ions. Among all the catalyst systems, DEAP-MCM-41 and MAP-MCM-41 (Table 3) catalysts found most efficient with toluene. Although use of toxic solvent and conventional approach limits the scope of method, but the variety of catalysts systems that have been investigated for the Friedlander reaction added catalyst variations.Garella et al. (2009) proposed solvent free microwave assisted Friedlander heteroannulation using propylsulfonic acid bounded silica as a catalyst [86]. The catalyst activity was performed under both the microwave (200W) as well as thermal conditions; microwave assisted protocol proved most efficient with shorter reaction times and higher yields. Varied yields of polysubstituted quinolines were reported. The catalyst recyclability was described for two more subsequent cycles without appreciable loss in catalytic activity.L\u00f3pez-Sanz et al. (2012) reported inorganic-organic hybrid materials based on SBA-15 molecular sieves as a catalyst for the Friedlander annulation [87]. Different catalyst systems based on SBA-15 were evaluated in toluene among them trialkoxysilyl 3-(propylamino) propane-1-sulfonic acid bounded on SBA-15 , Table 3) found most efficient. The article contributed new genre of hybrid catalyst although it involved lengthy catalyst preparation steps, use of toxic solvent.Smuszkiewicza et al. (2013) reported bifunctional mesoporous MCF materials as catalysts in the Friedlander condensation under solvent free conditions [88]. The MCF materials modified with (3-aminopropyl)trimethoxysilane; the silica MCF (AP/MCF), nio-biosilicate Nb-MCF (AP/Nb-MCF) and aluminosilicate Al-MCF were used as catalyst for Friedlander reaction. Among these, AP/Al-MCF seems comparatively most efficient with shorter reaction time and higher yields. The reusability of catalyst was also mentioned. The methodology shared new form of hybrid catalysts to the Friedlander synthesis. The same group has been reported the nio-biosilicate Nb-MCF with [3-(2-aminoethylamino) propyl] trimethoxysilane (2APMS), noted as 2APMS/NbMCF. Along with 2APMS/NbMCF, AP/MCF both in activated and nonactivated form were used as a catalyst. Nonactivated samples were found to be more efficient catalysts than the activated form [89].Ricciardi et al. (2013) demonstrated perfluoroalkylsulfonic acid monolayer-functionalized microreactor as a heterogeneous catalyst for the Friedlander annulation [90]. The 2-aminoacetophenone and ethyl acetoacetate in dry acetonitrile were passed through the continuous flow microreactor; affording excellent yields. The method added an innovative small-scale approach for the Friedlander reaction which involved steady inner acid functionalized surface of microreactor as a catalyst.S-proline sulphonamide as a novel hybrid heterogeneous catalyst [91]. The procedure involved use of water along with 2-aminobenzaldehyde, corresponding ketone and the hybrid catalyst. The recyclability for four consecutive cycles was mentioned with almost negligible loss in catalytic activity and enantioselectivity. Ba\u00f1\u00f3n-Caballero et al. (2013) investigated solvent free enantioselective Friedlander heteroannulation using silica-supported binam-3(BTC)2, Table 3] is a 3D porous MOF with a zeolite-like structure and which is commercially available as Basolite C300. Along with Basolite C300, two different molecular sieves, such as H-BEA and the mesoporous material (Al)SBA-15 were also employed as a catalyst. [Cu3(BTC)2] showed highly improved catalytic activity as compared with the molecular sieves, H-BEA and (Al)SBA-15. The same research group has reported one more paper using CuBTC MOF as efficient catalyst [93]. The article added new version of hybrid catalyst to the advancement in the Friedlander annulation. P\u00e9rez-Mayoral and \u010cejka (2011) proposed Friedlander synthesis using a metal-organic framework (MOF) based hybrid materials as a catalyst under solvent free conditions [92]. The [CuPhan et al. (2013) attempted modified Friedlander quinoline synthesis using CuBDC as a MOF based hybrid heterogeneous catalyst [94]. The modified Friedlander reaction involved use of 2-aminobenzylalcohol with corresponding ketone was treated in the presence of CuBDC (Table 3) catalyst; high amount of yield obtained. The recovered catalyst was reused for several times without significant loss in catalytic activity. Although, the MOF catalyst proved itself as efficient but, its preparation and characterization seem costlier.Jida and Deprez (2012) attempted Friedlander synthesis of polysubstituted quinolines and naphthyridines using propylphosphonic anhydride as a catalyst [95]. The reaction involved use of T3P (50% in ethyl acetate) as a promoter, after the completion of reaction water was added to the reaction mixture which dissolves the T3P and provides simple work up procedure. Excellent yields of substituted quinolines and naphthyridines were mentioned. The article has been successfully added a new form of catalyst with rich library of quinoline scaffolds.Pol60, Table 3) as a hybrid catalyst using Friedlander heteroannulation [96]. The ethanolic mixture of Friedlander reaction including TPAPol60 catalyst was refluxed; affording excellent yields of corresponding quinolines. The catalyst reusability was mentioned with consistent catalytic activity. Although the methodology seems conventional with longer reaction times; it has been found to be added a new form of hybrid catalyst to the improvement in the Friedlander reaction. Bennardi et al. (2015) employed tungstophosphoric acid included in a polymeric matrix of polyacrylamide demonstrated solvent free Friedlander quinoline synthesis using biodegradable sulfonated cellulose and starch as a solid acid catalyst [97]. Both the sulfonated cellulose and starch proved as an efficient biocatalyst, furnished good to excellent amounts of products but sulfonated cellulose (Table 4) found comparatively more efficient with shorter reaction times and higher yields. The article introduced new natural polymer supported catalyst for the advancement in the Friedlander annulation. The article limits the credit with lack of reusability of catalyst and high temperature conditions.Siddiqui and Khan (2013) achieved solvent free synthesis of benzopyranopyridines using chitosan (Table 4) as a biodegradable, heterogeneous catalyst via Friedlander cyclocondensation [98]. The catalyst activity was evaluated under both in refluxing methanol and solvent free conditions; solvent free conditions gave higher yields in shorter reaction times. The reusability of the catalyst for five consecutive cycles was mentioned with negligible loss in activity. Considerably short reaction times, solvent free condition, high yields, efficient reusability of catalyst and environment friendly catalyst makes the impressive contribution to the Friedlander advancements. 3H, Table 4) as a biodegradable solid acid catalyst [99]. The ethanolic reaction mixture of Friedlander components including chitosan-SO3H catalyst was reflux; affording high to excellent yields of products. The recovered catalyst was recycled for three subsequent cycles with negligible loss in catalytic activity. Although the methodology seems conventional but the short reaction times, efficient reusability of catalyst and use of biodegradable catalyst make the creditable improvement to the Friedlander condensation.Reddy et al. (2013) investigated synthesis of quinoline by Friedlander approach using chitosan sulfonic acid as a magnetically recoverable core-shell nanocatalyst for the Friedlander quinoline synthesis under solvent free conditions [100]. Excellent yields of polysubstituted quinolines were obtained. The recovered Fe3O4@SiO2/ZnCl2 nanocatalyst was recycled for five consecutive cycles with consisted activity up to second run and thereafter it decreases considerably. Economic yields, solvent free and low temperature conditions, magnetic recovery of catalyst, reusability of catalyst seems the valuable contribution of method; though it involved multistep preparation of catalyst.Soleimani et al. (2017) reported ZnCl3O4@SiO2-SO3H as a recyclable heterogeneous nanomagnetic catalyst [101]. Variety of active methylene compounds were used; affording high to excellent amounts of corresponding quinolines. The magnetically recovered nanocatalyst was reused for five subsequent cycles with gradual decrease in catalytic activity after each rerun. The catalyst efficiency was compared with earlier reported protocols. Although the method involves high temperature condition, but high yields, solvent free conditions, simple recovery and efficient reusability of catalyst seems advantageous.Beyki and Fallah-Mehrjardi (2017) demonstrated solvent free Friedlander synthesis of polysubstituted quinolines using Fe2O3 nanoparticles as a magnetic catalyst . The catalyst preparation involved sequence of steps. Excellent amounts of products and reusability of nanocatalyst for several ten more consecutive cycles without appreciable loss in activity was reported. The efficiency of [\u03b3-Fe2O3-HAp-(CH2)3-NHSO3H] nanocatalyst at the room temperature found remarkable part of the protocol. Sheykhan et al. (2011) [102] reported solvent free room temperature Friedlander heteroannulation using sulfamic acid heterogenized on hydroxyapatite encapsulated \u03b3-Fe3O4@SiO2 bonded N-propyl diethylene tetrasulfamic acid as a super paramagnetic nanocatalyst [103]. The method provided good to excellent amounts of corresponding quinolines. The recovered catalyst was reused for eight subsequent cycles with negligible loss in activity after each rerun. The protocol has been found to be added a novel magnetic nanocatalyst successfully, but the preparation of catalyst involved many steps which limits the credit of the methodology.Nasseri et al. (2014) attempted solvent free Friedlander heteroannulation using Fe3O4@SiO2-APTES-TFA, Figure 10A, Table 5) as a magnetic nanocatalyst for the solvent free Friedlander quinoline synthesis [104]. Excellent yields of polysubstituted quinolines were reported. Magnetically recovered catalyst was reused for four subsequent runs without significant decrease in the catalytic activity. The article introduced new hybrid magnetic nanocatalyst with efficient catalytic performance though it involved multistep preparation of catalyst and longer reaction times.Jafarzadeh et al. (2015) validated trifluoroacetic acid-aminopropyl triethoxysilane immobilized on silica coated magnetite nanoparticles was immobilized in both the form nano H3PMo12O40 (PMAn), and bulk H3PMo12O40 (PMAb) on the Fe3O4@SiO2-imid nanoparticles; both the form of catalysts showed excellent efficiency but Fe3O4@SiO2-imid-PMAn found more efficient with higher yields. The recovered nanocatalyst was recycled reused for four times without noticeable decrease in catalytic activity. The method added a new hybrid nanocatalyst though it involved lengthy preparation of catalyst.Esmaeilpour and Javidi (2015) demonstrated synthesis of polysubstituted quinolines via Friedlander heteroannulation using Fe3O4-Cys, Figure 10C, Table 5) as a magnetic nanocatalyst under solvent free conditions [106]. The efficiency of Fe3O4-Cys nanocatalyst was compared to the earlier reported catalysts protocol. The methodology added a new efficient hybrid magnetic nanocatalyst to the Friedlander advancement with short reaction times, excellent yields and recyclability of the catalyst for ten consecutive cycles was mentioned with no significant loss in catalytic activity.Bankar and Shelke (2018) reported microwave assisted Friedlander synthesis using ferrite-L-cysteine as a magnetic heterogeneous nanocatalyst under solvent free conditions [107]. Along with Friedlander reaction, the article also described the use of Nanocat-Fe-OSO3H as a catalyst for Ritter reaction and a multicomponent reaction. The model reaction was performed using Nanocat-Fe-OSO3H as a catalyst in short time under microwave irradiation; affording high yield. The reusability of magnetically recovered catalyst was mentioned for five subsequent cycles with negligible loss in catalytic activity. Gawande et al. (2013) reported microwave assisted Friedlander heteroannulation using magnetite-sulfonic acid nanoparticles (Nanocat-Fe-OSO2O4 nanoparticles (Table 5) as a nanocatalyst in aqueous medium [108]. The excellent yields of corresponding quinolines were obtained in short times. The recovered catalyst was reused for five successive cycles without any significant loss of activity. New form of efficient magnetic nanocatalyst was introduced to the catalyst library for Friedlander improvements. Baghbanian and Farhan (2014) achieved synthesis of quinolines via Friedlander reaction using CuFe2/Fe3O4-MNPs, Table 5) as a nanocatalyst in ethanolic medium [109]. The efficiency of the ZrO2/Fe3O4-MNPs nanocatalyst was compared with earlier reported catalysts protocols. The recovered catalyst was recycled for four consecutive cycles without noticeable loss in activity. Although the protocol seems conventional but short reaction times, excellent yields and efficient reusability of catalyst highlights the advantages of method. Hejazi et al. (2015) investigated Friedlander quinoline synthesis using zirconia supported on ferrite magnetic nanoparticles as a nanocatalyst in refluxing chloroform [110]. Excellent amounts of yields were reported. The catalyst reusability for three successive runs was mentioned with gradual decrease in activity. The methodology found to be added a new catalyst but, use of solvent, longer reaction times makes it align to conventional.Sadjadi et al. (2009) performed synthesis of polysubstituted quinolines via a Friedlander reaction using nanocrystalline alumina employed nanoporous cage-type aluminosilicate AlKIT-5 (Table 6) as a nanocatalyst for the synthesis of polysubstituted quinolines via Friedlander heteroannulation in refluxing ethanol [111]. The excellent yields of corresponding quinolines were reported with reusability of catalyst for three subsequent cycles with considerable decrease in catalytic activity after each cycle. The article introduced an efficient aluminosilicate mineral based nanocatalyst though the method seems conventional.2, Table 6) as a nanocatalyst [112]. The varieties of active methylene compounds were used for quinoline synthesis; high to excellent yields were obtained. The same silica nanocatalyst was also used for synthesis of quinoxalines efficiently. The reusability for fourteen more subsequent cycles with slight decrease in activity after each rerun was mentioned.Hasaninejad et al. (2012) demonstrated solvent free microwave assisted Friedlander annulation using silica nanoparticles HSO4/MCM-41] catalyst was compared with earlier reported polymeric catalysts protocol. The reusability of catalyst was unseen in the article. The solvent free reaction, short reaction times and economic yields seems the improvement over the classical Friedlander reaction.Abdollahi-Alibeik and Pouriayevali (2012) studied application of nanosized MCM-41 supported n-butanesulfonic acid pyridinium hydrogensulfate . Good to high amount of corresponding quinolines were obtained. The paper introduced an efficient silica mineral based nanocatalyst for the advancement of Friedlander reaction.Tahanpesar et al. (2014) reported solvent free Friedlander quinoline synthesis using ferric chloride supported on nanosilica . The variety of active methylene compounds were employed; furnished excellent amount of yields in short reaction times. The reusability of catalyst for five more consecutive cycles were performed and showed almost retained catalytic activity. The efficiency of catalytic activity was compared with more than 15 earlier reported catalytic protocols. Azimi and Abbaspour-Gilandeh (2014) demonstrated solvent free synthesis of polysubstituted/polycyclic quinolines via a solvent free Friedlander synthesis using Li2, Table 6) of different sizes as a heterogeneous catalyst for the Friedlander synthesis of quinolines under solvent free conditions [116]. The nano-TiO2 of size 16, 35, 70, 200 and 1000 nm were employed as a catalyst for the reaction; 200 and 1000 nm titania particles were inefficient as a catalyst while 16 nm size nano-TiO2 found the most efficient. The reusability of catalyst for four subsequent cycles was mentioned with gradual decrease in activity after each cycle. Bandyopadhyay et al. (2014) attempted titania nanomaterials (nano-TiOTeimouri and Chermahini (2016) performed Friedlander synthesis using nanocrystalline sulfated zirconia as a heterogeneous catalyst in refluxing ethanol [117]. Alongwith SZ, montmorillonite K-10 (K-10) and zeolite (ZMS-5) was also screened as catalyst for the reaction; all the catalyst systems proved almost equally efficient. The reusability of all the catalyst forms were reported for three successive cycles with gradual decrease in activity though SZ showed maximum efficiency. High to excellent yields of corresponding quinolines were mentioned in short reaction times.3, Table 6) nanoparticles as a nanocatalyst under solvent free conditions [118]. The model reaction was performed using nano-CaSiO3 catalyst. The article described the synthesis of calcium silicate nanoparticles using tetraethyl orthosilicate and soluble calcium nitrate tetrahydrate. The reusability of catalyst for four consecutive cycles was mentioned with gradual decrease in activity after third cycle. The library of quinolines was unseen in the article but, the article has successfully introduced a new catalyst to the advancement in the Friedlander heteroannulation.Palaniraja et al. (2017) synthesized polysubstituted quinolines via Friedlander reaction using wollastonite studied application of CuO nanoparticles as a nanocatalyst for the solvent free Friedlander heteroannulation [119]. Different metal oxides such as TiO2, ZnO, MgO, CaO, commercial ZnO (CM-ZnO) and NF-ZnO were screened as catalyst for the reaction; among all CM-ZnO and NF-ZnO found efficient however NF-ZnO proved comparatively most efficient with short reaction times and higher yields. The article comprises rich library of quinoline scaffolds prepared from different active methylene compounds. The catalyst NF-ZnO was reused for two more subsequent cycles with almost consistent catalytic activity.Hosseini-Sarvari (2011) reported Friedlander synthesis of polysubstituted quinolines using nanoflake as a nanocatalyst under solvent free conditions [120]. The various metal oxides such TiO2 nanoparticles as a catalyst via a modified Friedlander synthesis under solvent free conditions [121]. The reaction mixture containing substituted amino ketone 48, active methylene compound 49, a drop of conc. H2SO4 and the catalyst SnO2 nanoparticles was microwave (500 W) irradiated or ground by mortar and pestle; affording good yields of corresponding acridine derivatives 50 (Scheme 17). The microwave protocol took shorter time but under grinding conditions yields were comparatively high. The article reported hemolytic activity study of synthesized polycyclic quinolines.Roopan and Khan (2010) investigated synthesis of biologically active 9-chloro-6,13-dihydro-7-phenyl-5H-indolo -acridine derivatives using SnOVenkanna et al. (2014) performed synthesis of quinoline-2,3-dicarboxylates using nano-CuO powder (Table 6) as a catalyst in acetonitrile by modified Friedlander heteroannulation [122] (Scheme 18). Although the method seems conventional with use of solvent, longer reaction times but found to be reported high to excellent yields. The catalyst recovered was recycled for three consecutive cycles with gradual loss in activity after each cycle.3)2, Table 6] nanofiber mats as a heterogeneous catalyst [123]. High to excellent amount of yields were reported. The catalyst reusability for four more successive cycles was mentioned with almost retained catalytic activity. The article contributed a new form of hybrid catalyst for the improvement in the Friedlander reaction. The use of toxic solvent limits the credit of protocol and makes it align to conventional.Ziyadi and Heydari (2014) attempted Friedlander quinolines synthesis using ferric nitrate supported polyvinylalcohol [PVA/Fe demonstrated Friedlander synthesis using metal-doped carbon aerogels as a catalyst under solvent free conditions [124]. The different carbon aerogels were synthesized by polymerization of resorcinol (R) and formaldehyde (F) doping with transition metal nanoparticles. The nanocarbons doped with Co(0) and Cu(0) named RFCoS and RFCuS (Table 6), respectively. The catalyst systems were found to be efficient and affording high to excellent yields. The reusability of RFCuS catalyst was mentioned for two more subsequent cycles with consistent catalytic activity.2, Figure 11, Table 6) as a heterogeneous catalyst via a modified Friedlander heteroannulation under solvent free conditions [125]. The mixture of aminoketones 51, alkynes 52, and PPInCl-nSiO2 catalyst was heated solvent free conditions; affording high to excellent yields of substituted quinolines 53 (Scheme 19). The catalyst was reused for four consecutive cycles with consistent catalytic activity. The article shared new quinoline scaffolds and added PPInCl-nSiO2 as an efficient nanocatalyst to the Friedlander advancements. In spite of lengthy catalyst preparation, the protocol contributed to the literature.Azizi et al. (2018) reported synthesis of quinolines and pyrido quinolines catalyzed propylphosphonium tetrachloroindate ionic liquid supported on nanosilica described nanopalladium catalysed modified Friedlander quinoline synthesis under conventional conditions [126]. The mixture of 22 were coupled with primary alcohols 54 via hydrogen autotransfer process; yielded \u03b1-alkylated ketones 55 which further reacted with 2-aminobenzyl alcohols 21 via the modified Friedlander synthesis to afford polysubstituted quinolines 56 in moderate to good yields (Scheme 20). The reusability of catalyst was found inefficient due to considerable decrease in yield. The article proposed a modified Friedlander synthesis using nano-Ag-Pd/C catalyst though it involved use of toluene.Chen et al. (2013) achieved synthesis of quinolines through one pot, two step reaction using Ag-Pd alloy nanoparticles supported on carbon as a nanocatalyst [127]. The ketones Aegle mearmelos Correa aqueous leaf extract. The catalyst was reused for five more consecutive cycles with retained activity up to third cycle and thereafter it decreases considerably. The short reaction times, excellent yields, solvent free conditions, catalyst reusability highlights the advantages of the method over conventional Friedlander reaction.Angajala and Subashini (2015) reported solvent free Friedlander heteroannulation using nickel nanoparticles as a nanocatalyst [128]. The biosynthesis of nickel nano catalyst was achieved using literature method reported by the same group from 2CO3) and activated by steam (S); labelled namely RFS, RFNa, and RFNaS, respectively. Also other carbon aerogels were prepared as RFS-PO and RFS-PS . The model Friedlander reaction was performed using all the prepared carbon aerogel-based catalyst systems such as RFS, RFS-PO, RFS-PS, RFNaS, RFNa-PO, RFNa-PS; among all RFS-PO, RFS-PS (Table 6) showed most efficient catalytic activity. The authors reported metal free synthesis of quinolines using carbon aerogels; microwave assisted methodology taken shorter time than thermal.Godino-Ojer et al. (2017) attempted solvent free Friedlander quinoline synthesis using carbon aerogels as a nanocatalyst under thermal as well as microwave conditions [129]. The two organic RF aerogels were prepared using resorcinol (R) and formaldehyde (F) with or without polymerization catalyst (Na\u00a0forms, structure and nature\u00a0within the\u00a0development of Friedlander heteroannulation for the synthesis of novel quinoline scaffolds was reported\u00a0in the\u00a0literature. Around hundred articles are reviewed\u00a0during this\u00a0context which has involved\u00a0the utilization\u00a0of\u00a0sort of\u00a0polymers as a support to the reagent or catalyst support for the Friedlander reaction. Numerous attempts were noted, which\u00a0are\u00a0successfully contributed the remarkable modification\u00a0also\u00a0as advancement\u00a0for the\u00a0Friedlander reactions over the classical\u00a0and traditional\u00a0approaches. The few of research groups have successfully introduced synthesis\u00a0of latest\u00a0quinoline scaffolds via Friedlander cyclo-condensation with improved methodology.\u00a0The massive\u00a0number of protocols employed solvent free conditions, unconventional energy sources\u00a0like\u00a0a microwave irradiation, an ultrasonication; which have reported higher yields of desired poly-substituted quinolines. Polymers in hybrid forms as a catalyst were also reported with an efficient catalytic activity and reusability. Recently, in last decade; nanocatalysis has profoundly contributed the noticeable improvement\u00a0over\u00a0classical Friedlander reaction conditions with recyclability of catalyst especially with magnetic nanocatalyst and with simple\u00a0workup\u00a0procedures. Although, nanocatalysis proved itself as new and effective methodology; but, the preparation of nanocatalyst and its characterization seems costlier compare to\u00a0another\u00a0heterogeneous catalysts\u00a0which can\u00a0limit\u00a0the precious\u00a0impression of catalytic advancement. From the available literature, the advancements\u00a0made up for\u00a0Friedlander reaction which are remarkably noteworthy include simple heterogeneous\u00a0workup\u00a0protocol, reusability of polymeric catalyst, microwave assisted and solvent free conditions.The role of polymers\u00a0in several\u00a0field of polymer supported synthesis are challenged to develop new polymeric catalysts\u00a0which may\u00a0operate under moderate,\u00a0low temperature\u00a0reaction conditions with maximum productivity and recyclability. As per\u00a0this\u00a0literature,\u00a0this might\u00a0be\u00a0the sole\u00a0review\u00a0which can\u00a0highlight the role of polymers as a catalyst or as a catalyst or reagent support for the Friedlander heteroannulation;\u00a0which can\u00a0surely form\u00a0an upscale\u00a0and valuable resource for seekers in polymer supported synthesis of other biologically important heterocyclic scaffolds. Also, this review\u00a0will\u00a0help to encourage the researchers to develop greener synthetic approaches\u00a0and may provide newer ideas to look\u00a0and use\u00a0new\u00a0sorts of\u00a0nanocatalysts. The broader application of suitable polymer supports\u00a0can\u00a0also be\u00a0considered to\u00a0achieve total synthesis of pharmacologically active natural heterocyclic motifs which have challenged\u00a0upcoming researchers\u00a0in the\u00a0field.The researchers\u00a0working in the"} +{"text": "F344 rats were given saline, vitamin A placebo or vitamin A analogues orally for 4 consecutive days. The following day they were killed and their alveolar macrophages (AM phi) were harvested by lavage. The functional integrity of the AM phi was determined by their capacity to phagocytize opsonized SRBC and to kill syngeneic adenocarcinoma cell lines nonspecifically. Results showed that 4 days treatment with greater than 100 IU of vitamin A as retinyl palmitate per gram body weight rendered the AM phi tumoricidal against syngeneic mammary adenocarcinoma cell lines (MADB-100 and MADB-200) and that AM phi activated with retinyl palmitate showed increased ability to phagocytize opsonized SRBC. Other retinoids, such as retinoic acid and retinol, had the same effect of inducing tumoricidal activity in rat AM phi. AM phi harvested from normal rats were also rendered tumoricidal by direct interaction with greater than 10(3) IU ml-1 of retinyl palmitate for 24 h in vitro. Thus, vitamin A at high doses can increase the phagocytic and tumoricidal activities of rat AM phi."} +{"text": "Sixty patients with advanced breast cancer unresponsive to tamoxifen have been randomised to receive four course of mitozantrone, 14 mg m-2 (n = 30) intravenously every 3 weeks or megesterol acetate, 160 mg bd (n = 30). One in three patients (11 from each group) had substantial disease control for a minimum period of 6 months i.e., lack of progression; seven patients (23%) showed objective response to mitozantrone compared to four (13%) receiving megesterol. Non-progressive disease occurred in all sites, including visceral metastases and receptor negative patients. There were no significant differences between treatment groups in the median time (5 months each) to disease progression response duration or survival from commencing second-line therapy. Toxicity was considerably higher in the mitozantrone group. Second-line hormonal therapies can produce similar therapeutic results as those achieved from a short course of a 'short option' single agent cytotoxic in patients who were previously thought hormone insensitive. Provided that the patient does not have life threatening disease a trial of megesterol acetate is worth consideration in that it does not prejudice subsequent response to combination cytotoxic chemotherapy."} +{"text": "Regional chemotherapy allows further exploitation of the steep dose response curve of most chemotherapeutic agents, while systemic toxicity remains tolerable. We investigated the difference in maximally tolerated dose, pharmacokinetics and antitumour effect comparing administration of melphalan as a bolus in isolated liver perfusion (ILP) or via hepatic artery infusion (HAI). For these in vivo studies an experimental model for liver metastases in male WAG/Ola rats is obtained by subcapsular inoculation of CC531 rat colon carcinoma cells. In this system, ILP allowed administration of a two times higher dose than HAI (12 mg kg-1 vs 6 mg kg-1). In both treatment modalities systemic toxicity (leukopenia) was dose limiting. No hepatic toxicity was observed. Bolus administration of the maximally tolerated doses of melphalan in HAI (6 mg kg-1) and ILP (12 mg kg-1) resulted in four times higher concentrations in both liver and tumour tissue of the ILP treated rats. However, the ratio of mean drug concentration in liver vs tumour tissue appeared to be 1.5 times that found for HAI. In the range of the in tumour tissue measured melphalan concentrations the CC531 cells showed a steep dose response relationship in vitro. Whereas HAI resulted in significant tumour growth delay, complete remissions were observed in 90% of the rats treated with ILP. This study shows that with 12 mg kg-1 melphalan in ILP highly effective drug concentrations are achieved in CC531 tumour tissue; although the melphalan concentration in liver tissue shows an even higher increase than in tumour tissue, hepatic toxicity is negligible in this dose range.(ABSTRACT TRUNCATED AT 250 WORDS)"} +{"text": "We have detected somatostatin receptors (SSR) by autoradiography in 3/4 established small cell lung cancer (SCLC) cell lines but not in two non-SCLC cell lines. The growth of 1/3 SSR positive SCLC cell lines was significantly inhibited by the long-acting somatostatin analogue octreotide 10(-9) M. We treated 20 SCLC patients with octreotide 250 micrograms three times daily for 1 week prechemotherapy (six patients) or at relapse after chemotherapy (14). Octreotide was well tolerated, and serum insulin-like growth factor-I levels were suppressed to 62 +/- 7% of pre-treatment levels. However there was no evidence of anti-tumour activity measured by tumour bulk or serum levels of neuron-specific enolase. In one patient metastatic skin nodules were shown to be SSR positive before and at the end of 2 weeks octreotide. Despite this the patient had progressive disease, and tumour cells obtained by fine needle aspirate before and after treatment showed no growth inhibition when cultured with octreotide immediately or following establishment as a cell line. In summary we saw little correlation between SSR expression and growth inhibition by octreotide, either in vitro or clinically."} +{"text": "Forty-one patients receiving remission induction chemotherapy with vincristine, adriamycin and prednisolone (VAP) for high grade lymphoma or acute lymphoblastic leukaemia were entered into a double blind, placebo controlled trial of oral acyclovir prophylaxis against herpes simplex virus (HSV) infection. The dose of acyclovir was 200 mg four times daily for the duration of chemotherapy (six weeks). Of the 40 evaluable patients, 20 were randomised to each arm. Prophylactic oral acyclovir significantly reduced the incidence of clinical HSV infection from 60% on placebo to 5% acyclovir (P less than 0.001), and the incidence of viral isolates from 70% on placebo to 5% on acyclovir (P less than 0.001)."} +{"text": "Twenty patients with epithelial ovarian carcinoma were treated with high-dose cisplatin 200mg m-2. Patients were to receive three cycles at 21 day intervals. Treatment was stopped if severe myelosuppression or any neurotoxicity occurred. Overall, eight (40%) of patients responded with a complete response in five (25%). Four of 16 (25%) previously treated patients responded. The median duration of response was 44 weeks (range 6-130). In patients previously treated there was a significant association (P < 0.002) between response and a remission free interval of 52 weeks or more from primary chemotherapy. Toxicity was assessable in 18 patients. Alopecia and nausea/vomiting were common. Myelosuppression was recorded in nine patients delaying planned administration in eight of 35 cycles. Five patients developed anaemia and six thrombocytopenia. Neurotoxicity affected seven patients and varying degrees of tinnitus six patients. Neurotoxicity and myelosuppression were indications for cessation of treatment in 8 patients receiving less than three cycles. Analysis revealed no significant association between toxicity and prior cisplatin exposure, age or the amount of high-dose cisplatin administered. This series reveals that it is possible to achieve good response rates using high-dose cisplatin without encountering debilitating neurotoxicity."} +{"text": "Sixty-nine unselected patients with locally advanced and metastatic carcinoma of the pancreas, who had not received previous chemotherapy or radiotherapy were randomised to receive either 5-fluorouracil, epirubicin and mitomycin C (FEM) or epirubicin. Survival was not significantly different in the two arms. Toxic reactions (WHO grade greater than 3) in the FEM and epirubicin arm respectively included nausea (2), (4), severe alopecia (1) (3) and leucopenia (1), (5), none of these were statistically significant. We therefore suggest that combination chemotherapy should not be used in preference to single agent chemotherapy as standard treatment for locally advanced or metastatic cancer of the pancreas."} +{"text": "Mitotane is considered to be the drug of choice for patients with inoperable, recurrent and metastatic adrenocortical carcinoma, although a favourable effect of this drug on survival has never been documented. We evaluated the efficacy of mitotane treatment of 96 patients with adrenocortical carcinoma followed up in our department between 1959 and 1992. Complete tumour resection was the goal of the initial treatment. Mitotane treatment was classified according to serum trough concentrations on maintenance therapy: low (< 14 mg l-1) or high (> or = 14 mg l-1). Total tumour resection was feasible in 47 patients (49%), and subtotal resection was performed in 37 patients (39%). Patients who underwent total tumour resection survived significantly longer than those who did not (P < 0.001). Adjuvant mitotane therapy (n = 11) did not influence survival after total resection. Sixty-two patients were given mitotane treatment at some time during their illness, only 30 of whom reached high maintenance serum levels. Mitotane treatment with high serum levels had an independently favourable influence on patient survival, using univariate (P < 0.01) and multivariate analysis (P = 0.01). Mitotane treatment resulting in low serum levels was tantamount to not giving mitotane at all. We conclude that mitotane treatment in adrenocortical carcinoma is effective only when high serum levels can be achieved."} +{"text": "The release of components from human kidney tumour xenografts (GYL) and human foetal kidney explants maintained in nude mice has been studied. The GYL tumour released antigens into the serum which could be detected by the generation of antibodies following cross-immunisation of closely related hairy litter mate (HLM) mice. The production of anti-GYL antibody was monitored by an I125 binding assay using viable GYL tumour cells. In 2/16 hairy litter mate mice, cell surface antibody binding by GYL cells was twice that found with 8 other human tumour cell lines (including 2 other kidney cancer cell lines). Absorption of these antisera with 10(7) GYL tumour cells completely abolished this response, where 50%, 38% and 25% of activity remained following absorption with; a normal kidney cell line, a homogenate of normal kidney and a mixed pool of human tumour cells. Six out of 8 GYL tumour bearing nude mice tested had elevated plasma levels of HCG. Absorption of the HLM antisera with an excess of commercial HCG abrogated I125 binding by only 15%, suggesting that antibody production was not directed primarily against ectopic HCG."} +{"text": "Survival data on a population-based series of bone, soft tissue and visceral sarcomas diagnosed in the North West of England between 1982-84 and subjected to histopathological peer review are presented. Five-year crude survival for all cases was 34%. Survival in males and females did not differ significantly but was markedly worse for patients diagnosed over the median age of 60 years, even when allowance was made for underlying mortality . Five-year survival rates for the major site groups were: bone 44%; soft tissues of head, neck and trunk 36%; soft tissues of extremities 35%; female genital tract 35%; retroperitoneum 15%; gastro-intestinal tract 13%. Analysis by the major histological types revealed the following survival rates: leiomyosarcoma--female genital tract 25%, gastro-intestinal tract 14%, non-visceral soft tissue 21%; malignant fibrous histiocytoma of soft tissue 29%; liposarcoma 52%; osteosarcoma of bone 46%; and chondrosarcoma of bone 50%."} +{"text": "Serum samples from 62 patients with seminoma were assayed for placental alkaline phosphatase-like activity using the monoclonal antibody H17 E2, in order to evaluate its utility as a serum tumour marker. Fifteen of 16 patients (94%) with active seminoma had elevated serum PLAP levels. Sixteen of 46 (35%) of patients considered to be in remission had elevated PLAP levels . Fifteen false positive results were considered attributable to concomitant smoking, and if these patients are excluded, only one false positive case was detected. In 7 out of 7 patients sequential PLAP assays reflected clinical response to treatment."} +{"text": "Ge(IV) phthalocyanine (GePc) with two axially ligated cholesterol moieties was prepared by chemical synthesis and incorporated in a monomeric state into small unilamellar liposomes (CGP 55398). Upon photoexcitation with light wavelengths around its intense absorption peak at 680 nm, GePc shows an efficient photosensitising activity towards biological substrates through a mechanism which largely involves the intermediacy of singlet oxygen. GePc injected systemically into mice bearing an intramuscularly implanted MS-2 fibrosarcoma is quantitatively transferred to serum lipoproteins and localises in the tumour tissue with good efficiency: at 24 h post injection the GePc content in the tumour is 0.74 and 1.87 micrograms per g of tissue with a tumour/peritumoral ratio of 4.35 and 5.67 for injected doses of 0.76 and 1.52 mg kg-1 respectively. At this time the red-light irradiation of the GePc-loaded fibrosarcoma causes a fast and massive tumour necrosis involving both malignant cells and blood vessels."} +{"text": "Cough variant asthma (CVA) is a cause of chronic cough and a precursor of typical asthma. We retrospectively examined the longitudinal change in bronchial responsiveness and cough reflex sensitivity in CVA patients with respect to the effect of long-term inhaled corticosteroids (ICS).Provocative concentration of methacholine causing a 20% fall in forced expiratory volume in one second (PC20-FEV1) and provocative concentration of capsaicin eliciting 5 or more coughs (C5) were measured before treatment and during a follow up period following relief of cough years after the initial visit) in a total of 20 patients with CVA .Three of 8 patients not taking long-term ICS developed typical asthma compared to none of 12 patients taking ICS (p = 0.0171). PC20-FEV1 significantly (p < 0.0001) increased from 1.80 to 10.7 mg/ml in patients taking ICS but did not change in patients not taking ICS [2.10 compared to 2.13 mg/ml]. Cough threshold did not change in patients whether taking or not taking ICS.Long-term ICS reduces bronchial hyperresponsiveness in CVA as recognized in typical asthma. Cough reflex sensitivity is not involved in the mechanism of cough in CVA. Cough variant asthma is a well-known cause of chronic non-productive cough as well as gastroesophageal reflux-associated cough and post-nasal drip-induced cough .Pathophysiological features of cough variant asthma appear tJohnson reportedIn our previous study , long-teAlthough some researchers reportedWe examined longitudinal changes in bronchial responsiveness and cough reflex sensitivity and influence of ICS on both responses in patients with cough variant asthma. Bronchial responsiveness to methacholine and cough reflex sensitivity to inhaled capsaicin were measured at least two times; at the initial visit and during the follow up period after relief of cough on treatment.Twenty patients with cough variant asthma as a single cause of chronic cough , who had undertaken spirometry, bronchial reversibility test, methacholine provocation test, capsaicin cough provocation test, measurements of peripheral blood eosinophil count, serum total IgE and specific IgE to common allergens, and induced sputum eosinophil count at presentation, were followed up with special emphasis on typical asthma onset during 6 months or more is desirable because the long-term therapy is recommended by many asthma guidelines in typical asthma even if the disease severity is mild. Long-term treatment with ICS was accepted and taken by 12 patients but not by the other 8 patients.The diagnosis of cough variant asthma was made according to the following criteria proposed by Japanese Cough Research Society , excludi1) Isolated chronic non-productive cough lasting more than 8 weeks2) Absence of a history of wheezing or dyspnea, and no adventitious lung sounds on physical examination3) Absence of post-nasal drip to account for the cough4) Forced expiratory volume in one second (FEV1), forced vital capacity (FVC), and FEV1/FVC ratio within normal limits 5) Presence of bronchial hyperresponsiveness <10 mg/mL)6) Relief of cough with bronchodilator therapy7) No abnormal findings indicative of cough aetiology on chest roentgenogramAll patients with cough variant asthma had been successfully treated with bronchodilators and/or corticosteroids, without use of other medications such as proton pump inhibitors and histamine H1-antagonists. Thus, cough variant asthma was the single cause of chronic cough in all patients studied.The efficacy of bronchodilator therapy described above was assessed according to the following criteria:1) \"Excellent\" when cough totally resolved2) \"Good\" when sleep and daytime quality of life were improved3) \"Fairly good\" when severity and frequency of cough were somewhat decreased4) \"Poor\" when cough was unchangedAn assessment of \"Excellent\" or \"Good\" was judged as effective.Pulmonary function, cough reflex sensitivity, and bronchial responsiveness were measured in that order within two weeks of the first visit Table , and theThe onset of typical asthma was defined as wheezing and/or dyspnoeic attack responding to bronchodilator therapy.2 test. Changes within group were assessed using the paired t test. PC20-FEV1 and C5 were analyzed using logarithmically transformed values. A p value of 0.05 or less was considered significant.Data excluding PC20-FEV1 and C5 were presented as mean \u00b1 standard deviation (SD). PC20-FEV1 and C5 were expressed as geometric mean value with geometric standard error of the mean. Differences between groups were determined by parametric one-way analysis of variance (ANOVA) or the \u03c7Typical asthma onset was recognized in 3 (37.5%) of 8 patients not taking long-term inhaled corticosteroids (ICS) and none of 12 patients taking ICS. The prevalence of asthma onset was significantly (p = 0.0214) different between the groups. Details of the 3 patients who developed typical asthma are shown in Fig. Clinical parameters at initial presentation are summarized in Table PC20-FEV1 significantly (p < 0.0001) increased by 5.9 GSEM, 1.40) times from 1.80 GSEM, 1.35) to 10.7 mg/ml in patients taking ICS, but did not change in patients not taking ICS [by 0.97 times from 2.13 to 2.10 mg/ml] (Fig. Cough variant asthma was first described by Glauser . The onlNearly 30% of cough variant asthma patients eventually develop wheezing, sometimes severe enough to require continuous treatment with bronchodilators . In thisThe present study clearly showed that bronchial responsiveness did not change after relief of cough without use of ICS, and long-tem ICS attenuated bronchial responsiveness to inhaled methacholine in patients with cough variant asthma, probably resulting in prevention of development of typical asthma from cough variant asthma. There were only 3 patients developing typical asthma whose bronchial responsiveness was more increased among the patients not taking ICS and was not obviously increased after the asthma onset as shown in Fig. It has been shown that early induction of ICS within 2 years following asthma onset is beneficial to achieve both control of symptom and improvement of pulmonary function and bronchial responsiveness in asthma . AlthougAlthough other researchers have reported that cough reflex sensitivity was heightened and recovered to a normal level following successful treatments of cough variant asthma -23, it sEarly induction of ICS within 2 years following asthma onset has been shown to be beneficial in attenuating bronchial hyperresponsiveness as well as achieving both control of symptom and improvement of pulmonary function . In thismay prevent onset of typical asthma from cough variant asthma by reducing bronchial hyperresponsiveness, and that cough reflex sensitivity is not involved in mechanism of cough in cough variant asthma. Further studies including randomized placebo-controlled studies are needed to confirm the preventive effect of long-term ICS on typical asthma onset from cough variant asthma.The present retrospective study showed that bronchial hyperresponsiveness and cough reflex sensitivity did not change after relief of cough when ICS therapy was not taken in patients with cough variant asthma. A median of 2 years ICS treatment attenuated bronchial hyperresponsiveness, but not cough reflex sensitivity. Bronchial responsiveness did not further increase after onset of typical asthma in 3 patients not taking ICS. These findings suggest that long-term ICS treatment ANOVA = analysis of variance, C5 = provocative concentration of capsaicin eliciting 5 or more coughs, CVA = cough variant asthma, FEV1 = forced expiratory volume in one second, FVC = forced vital capacity, GSEM = geometric standard error of the mean, ICS = inhaled corticosteroids, PC20-FEV1 = provocative concentration of methacholine causing a 20% fall in forced expiratory volume in one second, SD = standard deviation,."} +{"text": "The aim of the present study was to analyse in invasive carcinomas of the uterine cervix, the anionic glutathione S transferase (GST pi) gene, possibly implicated in the drug resistance of human cancers. Total RNA preparations obtained from invasive cervical cancers (106 specimens), carcinomas in situ (CIS) (three specimens) and normal cervical epitheliums (24 specimens) were analysed by Northern and slot blot hybridisation. A 0.7 kb GST pi transcript band was detected in all the cervical specimens. GST pi mRNA levels were lower in normal cervix (mean: 0.7 +/- 0.1 arbitrary units) than in invasive carcinomas (mean: 2.5 +/- 1.5 units) (Student test P less than 10(-4)). However no significant difference was observed between invasive cancers of advanced stages (III and IV) and those of early stages (I and II). The presence of human papillomavirus in cancers and in normal cervices did not influence significantly the GST pi mRNA level. Neither amplification nor gross rearrangement of GST pi gene could be observed after Southern blot analysis of genomic DNA. In conclusion, our data indicate that the presence of high levels of GST pi transcripts in invasive cancers may be a consequence of the multiple biochemical changes which accompany cervical carcinogenesis."} +{"text": "Computed tomographic scanning of the chest in 100 patients with newly diagnosed malignant lymphoma detected mediastinal lymphadenopathy (39%) and parenchymal deposits (15%) with a significantly greater sensitivity and specificity than conventional radiological techniques. This principally affected the staging and treatment of patients with limited stage disease. The stage was changed in 10/61 patients (16%) with Stages I-III prior to CT scan and treatment was altered in 11/29 (38%) patients for whom radiation was the treatment of choice. Complete remissions as defined by CT scan have been more durable than those defined by CXR alone."} +{"text": "Flunarizine is a class IV calcium channel blocker which increases oxygen delivery to hypoxic regions in solid tumours, exerting a radiosensitising effect in vivo in animal tumour models. Precisely how the drug improves oxygenation is not well understood. We hypothesised that metabolic conditions present within solid tumours reduce red blood cell (RBC) deformability and that flunarizine exerts its in vivo effect by preventing this loss of RBC deformability. A microrheometer was used to compare the viscosity of rat and human RBC suspensions in conditions of hypoxia (pO2 < 10 mmHg), acidic environment (pH 6.8), and elevated lactate concentration (lactate 5 mMol l-1), without or with flunarizine at concentrations of 5, 10, and 50 mg l-1. The effects of flunarizine on RBC density and morphology were also recorded. Hypoxia, low pH, and lactate exposure together increased both human and rat RBC suspension viscosity. Flunarizine at concentrations of 5 and 10 mg l-1 prevented the increases in viscosity. The drug caused dose-dependent shifts toward lower cell density while inducing a characteristic cupped shape (stomatcytic morphology), suggesting a mechanism involving calmodulin inhibition. The results support the hypothesis that flunarizine improves tumour blood flow and oxygenation by enhancing flow properties of RBC's in solid tumours."} +{"text": "A radioimmunoassay for neuron specific enolase (NSE), a marker of neuroendocrine differentiation, has been evaluated in small cell lung cancer (SCLC). In untreated patients 25/38 (68%) with localized SCLC had raised blood levels of NSE (greater than 13 ng ml-1), in extensive disease 34/39 (87%) patients had raised NSE levels. In patients with non-small cell lung cancer (NSCLC) the serum levels were raised in 16/94 (17%). In extensive tumours of non-pulmonary origin NSE levels were increased in 24/116 (20%) patients. Longitudinal studies indicated a good correlation between the response to chemotherapy and fall of NSE levels. Tumour progression was accompanied by a rising NSE in 25/29 patients, with doubling times of 7-90 days. In patients with progression with a normal NSE the recurrence was a NSCLC. Cerebral metastases occurring as the only recurrence during clinical complete remission were not accompanied by a rise of NSE. Serum NSE levels provides a valuable monitor for SCLC during and after chemotherapy."} +{"text": "The relationship between hepatic arterial albumin microsphere distribution and hepatic arterial blood flow and the effects of regional angiotensin II were studied in a rat liver metastases model. Hooded-Lister rats were inoculated subcapsularly with 2 x 10(6) HSN sarcoma cells. At 20 days, hepatic arterial blood flow was measured using the reference microsphere technique. Animals then randomly received 50 microliters hepatic arterial saline or albumin microspheres . Hepatic arterial blood flow measurements were then repeated at 5 min. After 5 min, animals were killed and tissues were weighed and counted in a gamma well counter. There were no significant differences between the hepatic blood flow measurements recorded before and after the control hepatic arterial saline infusion. However, regional albumin microspheres produced a significant reduction in tumour and normal liver blood flow and an 80% reduction in mean T/N blood flow ratio. Regional albumin microspheres were delivered to tumour in greater proportions than would be expected from baseline hepatic arterial blood flow . There was no correlation between T/N for baseline blood flow and albumin microsphere distribution."} +{"text": "Photodynamic therapy consists of the systemic administration of a derivative of haematoporphyrin (Photofrin II) followed 24-72 h later by exposure of malignant lesions to photoradiation. We investigated the efficacy of this treatment after direct intratumoral injection of Photofrin II. This direct treatment regimen resulted in higher rates of inhibition of mitochondrial cytochrome c oxidase (5.13% J-1 cm-2 x 10(-1) and succinate dehydrogenase , respectively. A significant delay in tumour growth in vivo was observed in animals that received intratumoral injections 2 h before photoradiation compared to animals injected intraperitoneally at either 2 or 24 h before photoradiation. The treatment protocols were compared with control groups, consisting of Photofrin II administration intratumorally or intraperitoneally without photoradiation, or photoradiation in the absence of Photofrin II. These data indicate that the intratumoral injection regimen with Photofrin II enhanced the efficacy of photodynamic therapy. The greater delay in tumour growth observed after intratumoral administration of Photofrin II suggests a mechanism favouring direct cell damage."} +{"text": "Dose limiting systemic toxicity prevents sufficient exploitation of the steep dose response relationship of most anticancer agents. In our rat liver tumour model , isolated liver perfusion allows administration of higher doses of mitomycin C than hepatic artery infusion, while systemic toxicity remains minimal. To determine the temporal pattern of mitomycin C induced cytokinetic changes, we analysed flow cytometric DNA histograms of CC531 liver tumours from rats treated with high dose mitomycin C (3.2 mg kg-1) via hepatic artery infusion and sacrificed at different time intervals after treatment. Between 12 and 36 h after treatment, the fraction of cells in late S and G2/M phase had markedly increased. The effects of administration of the respective maximally tolerated doses of mitomycin C in isolated liver perfusion and via hepatic artery infusion on progression of tumour cells through the cell cycle and on gross tumour growth were compared. Isolated liver perfusion with mitomycin C resulted in a significant increase in the proportion of cells in mid and late S, and in some accumulation of cells in early S and G2/M phase at 24 and 48 h after treatment. In contrast, after hepatic artery infusion a significant increase of the fraction of cells in G2/M phase was observed at 24 h after treatment. Monitoring tumour growth after isolated liver perfusion five out of seven rats showed a complete tumour remission, while after hepatic artery infusion only a minimal growth delay was detected. This study demonstrates that isolated liver perfusion in the rat CC531 liver tumour model allows the administration of a well-tolerated dose of mitomycin C being high enough to induce a marked DNA synthesis inhibition and even complete tumour remission."} +{"text": "Proliferative activities in 181 primary Borrmann type 4 gastric carcinomas were investigated using percentage labelling of proliferating cell nuclear antigen (PCNA) and an argyrophilic nucleolar organiser region (AgNOR) count. Tumours with a high proliferative activity often metastasised to lymph nodes (P < 0.01), and these patients had a lower survival rate (P < 0.05). A significant correlation was recognised between the PCNA labelling percentage and AgNOR count . Cox's regression analysis showed that PCNA labelling percentage is an independent prognostic factor. These results indicate that estimating proliferative activity may be useful in predicting lymph node metastasis and patients' prognosis in cases of Borrmann type 4 gastric carcinoma."} +{"text": "Sarracenia purpurea. Path models that incorporated food-web structure better predicted population sizes of food-web constituents than did simple keystone species models, models that included only autecological responses to habitat volume, or models including both food-web structure and habitat volume. These results provide the first experimental confirmation that trophic structure can determine species abundances in the face of habitat loss.Plant and animal population sizes inevitably change following habitat loss, but the mechanisms underlying these changes are poorly understood. We experimentally altered habitat volume and eliminated top trophic levels of the food web of invertebrates that inhabit rain-filled leaves of the carnivorous pitcher plant Manipulating habitat volume and the presence of top trophic levels of an aquatic community reveals that trophic structure can determine species abundance despite habitat loss. The loss of natural habitat area often is accompanied by the disappearance of large-bodied top predators and the upper trophic levels of food webs \u20133. HowevFor multitrophic assemblages, two broad classes of community models predict the potential responses of populations to habitat change. (1) Single-factor models emphasize the unique responses of individual species to variation in habitat area. This framework includes island biogeographic models , as wellUnfortunately, published studies of the effects of habitat contraction have relied on conventional analyses that do not explicitly compare these alternative frameworks ,18. AlthSarracenia purpurea and sarcophagid fly (Fletcherimyia fletcheri) larvae [(Habrotrocha rosi) and mites (Sarraceniopus gibsonii). Larvae of the pitcher plant mosquito Wyeomyia smithii feed on bacteria, protozoa, and rotifers [F. fletcheri feed on rotifers and small (first and second instar) larvae of W. smithii [Sarracenia food web exhibits the same complex linkages across multiple trophic levels that characterize other aquatic and terrestrial food webs [S. purpurea throughout its broad geographic range\u2014from the Florida panhandle to Labrador and west to the Canadian Rocky Mountains [The macroinvertebrate community associated with the northern pitcher plant purpurea is a modd States . The plad States . Leaves d States . The bas) larvae . Shredde) larvae , which rrotifers . Large . We measured the abundance of the resident species in each replicate leaf and simplified the trophic structure of the ate leaf and compate leaf .Wyeomyia keystone model in the path models. In the first grouping Click here for additional data file.Protocol S1(192 KB DOC)Click here for additional data file.Table S1(31 KB DOC)Click here for additional data file.Table S2(34 KB DOC)Click here for additional data file.Table S3(34 KB DOC)Click here for additional data file.Table S4(33 KB DOC)Click here for additional data file.Table S5(33 KB DOC)Click here for additional data file.Table S6(33 KB DOC)Click here for additional data file.Table S7(33 KB DOC)Click here for additional data file.Table S8(33 KB DOC)Click here for additional data file.Table S9(34 KB DOC)Click here for additional data file."} +{"text": "Using conventional examination (CE) of H&E stained slides from bone marrow aspirates, metastases can be detected in approximately 25% of patients with small cell lung cancer. We investigated a panel of monoclonal antibodies using immunohistochemistry in the diagnosis of bone marrow infiltration from SCLC and compared the results with CE. Seven monoclonal antibodies raised against epithelial antigens were applied on bone marrow sections from three groups of patients (pts): (1) 19 pts in whom SCLC-metastases were detected by CE, (2) 44 pts with SCLC in whom metastases could not be detected by CE, and (3) 20 pts with non-malignant bone marrow diseases. All the antibodies except LCA1/L38 were positive in 60-90% of the slides with infiltrating tumour cells in group 1. No positive tumour cells were detected in group 2. A few plasma cells and megakaryocytes were slightly positive for MOV 15 and NCCST 433, but no other positive cells were detected in group 3. In conclusion, the monoclonal antibodies used in this study may be useful for diagnostic purposes when a suspicious looking infiltration is detected by CE. However, these antibodies could not detect metastatic tumour cells in the bone marrow sections from patients in whom CE did not reveal any tumour cells."} +{"text": "Twenty-six patients with treated breast cancer who had been randomised previously to receive combination chemotherapy including alkylating agents (n = 14) or to undergo oophorectomy (n = 12) following surgery underwent cytological and colposcopic screening of the uterine cervix. Colposcopically directed cervical punch biopsies were taken from all patients in whom a colposcopic abnormality was detected. Breast cancer patients were compared with 79 controls with normal cervical cytology and no known breast malignancy. Colposcopically directed punch biopsies were taken from the cervical transformation zone of all controls. Significantly more breast cancer patients who had received chemotherapy (43%) than controls (10%) had CIN (P < 0.01) and significantly more patients who had received chemotherapy (14%) than controls (3%) had CIN 2 or 3 (P < 0.05). The proportion of breast cancer patients in the oophorectomy group with CIN (17%) did not differ significantly from the control group. No case of CIN was detected by cervical cytology. This study suggests that breast cancer patients receiving combination chemotherapy including alkylating agents are at increased risk of CIN, and that cervical cytology alone may be an inadequate form of screening for these patients."} +{"text": "One hundred and seventy-eight patients with non metastatic inflammatory breast cancer (IBC) have been treated at the Centre H. Becquerel. Median follow up is 67 months (6-178). Every patient received neoadjuvant chemotherapy , followed by a loco regional treatment , followed by adjuvant chemotherapy. During this period, the types of chemotherapy and locoregional treatment have been the following: Study I: 64 patients treated with CMF or AVCF and XRT; Study II: 83 patients, treated with either AVCF, FAC or VAC followed by S (n = 38) or XRT (n = 22) in case of complete or partial response, or followed by XRT (23) in case of initial supraclavicular lymph node involvement or lack of response after chemotherapy; Study III: 31 patients treated with FEC-HD + Estrogenic recruitment followed by S and XRT after adjuvant chemotherapy, except seven patients who received XRT . Although objective response rates are statistically better in the 3rd study, this does not translate in dramatically different disease free survival or overall survival . Analysis of subset of patients without supra clavicular lymph node involvement where neoadjuvant chemotherapy obtained at least a 50% response reveals a median disease free survival and median overall survival of respectively 38.3 and 60.1 months for patients who underwent S vs 19 and 38.3 months for those who received XRT (P = 0.15). These studies suggest that surgery has no deleterious effect on outcome of IBC. Advantage on disease free survival or overall survival from intensive chemotherapy in IBC remains to be proven with appropriate randomised trials."} +{"text": "INK4a generates supernumerary centrosomes through centriole pair splitting. Generation of supernumerary centrosomes in human diploid epithelial cells was shown to nucleate multipolar spindles and directly drive production of aneuploid daughter cells as a result of unequal segregation of the genomic material during mitosis. Finally, we demonstrate that p16INK4a cooperates with p21 through regulation of cyclin-dependent kinase activity to prevent centriole pair splitting. Cells with loss of p16INK4a activity have been found in vivo in histologically normal mammary tissue from a substantial fraction of healthy, disease-free women. Demonstration of centrosome dysfunction in cells due to loss of p16INK4a suggests that, under the appropriate conditions, these cells can become aneuploid. Gain or loss of genomic material (aneuploidy) may provide the necessary proproliferation and antiapoptotic mechanisms needed for the earliest stages of tumorigenesis.Aneuploidy, frequently observed in premalignant lesions, disrupts gene dosage and contributes to neoplastic progression. Theodor Boveri hypothesized nearly 100 years ago that aneuploidy was due to an increase in centrosome number (multipolar mitoses) and the resultant abnormal segregation of chromosomes. We performed immunocytochemistry, quantitative immunofluorescence, karyotypic analysis, and time-lapse microscopy on primary human diploid epithelial cells and fibroblasts to better understand the mechanism involved in the production of supernumerary centrosomes (more than two microtubule nucleating bodies) to directly demonstrate that the presence of supernumerary centrosomes in genomically intact cells generates aneuploid daughter cells. We show that loss of p16 Here the authors show that aneuploidy can be caused by cells having an abnormal number of centrosomes, which leads to asymmetric cell division. Zur Frage der Entstehung maligner Tumoren , published in 1914, Theodor Boveri hypothesized that multipolar mitoses cause aneuploidy ) that express EGFP\u2013\u03b3-tubulin (green) and EGFP-H2B (green) during mitotic progression of cells that divide into two nuclei with two centrosomes . The (219 KB MOV).Click here for additional data file.Video S2x (green arrow), y (red arrow), and z (blue arrow) position designates the orientation of the cells (lower left corner). The red grid designates the proportional scale. The relative time of each frame is placed in the upper right corner.Time-lapse microscopy of early-passage vHMECs (RM18 [13 to 33 PD]) that express EGFP\u2013\u03b3-tubulin (green) and EGFP-H2B (green) during mitotic progression of cells that divide into two nuclei with more than two centrosomes . The (244 KB MOV).Click here for additional data file.Video S3x (green arrow), y (red arrow), and z (blue arrow) position designates the orientation of the cells (lower left corner). The red grid designates the proportional scale. The relative time of each frame is placed in the upper right corner.Time-lapse microscopy of early-passage vHMECs (RM18 [13 to 33 PD]) that express EGFP\u2013\u03b3-tubulin (green) and EGFP-H2B (green) during mitotic progression of cells that divide into greater than two nuclei with more than two centrosomes . The (97 KB MOV).Click here for additional data file.http://www.ncbi.nlm.nih.gov/Genbank) accession numbers for proteins discussed in this paper are p16INK4a (NM_000077), human centrin 2 (NM_004344), H2B (X00088), \u03b3-tubulin (NM_001070), and p21 (NM_078467).The GenBank ("} +{"text": "Cytosols of human breast tumours have been assayed for DNA dependent DNA polymerase alpha activity. DNA polymerase alpha activity in benign tumours was found to be significantly lower than in untreated malignant tumours. Biopsy samples removed surgically before and after endocrine therapy showed reduced DNA polymerase alpha activity in 6 out of 9 patients treated with 4-hydroxyandrostenedione, and in 6 out of 7 patients treated with MPA. DNA polymerase alpha activity in malignant breast tumours was higher in oestrogen receptor negative than oestrogen receptor positive tumours."} +{"text": "Easy Electroporation of Zygotes (EEZy) allows highly efficient generation of CRISPR/Cas9 transgenic mice while reducing the numbers of animals required.Electroporation of zygotes represents a rapid alternative to the elaborate pronuclear injection procedure for CRISPR/Cas9-mediated genome editing in mice. However, current protocols for electroporation either require the investment in specialized electroporators or corrosive pre-treatment of zygotes which compromises embryo viability. Here, we describe an easily adaptable approach for the introduction of specific mutations in C57BL/6 mice by electroporation of intact zygotes using a common electroporator with synthetic CRISPR/Cas9 components and minimal technical requirement. Direct comparison to conventional pronuclear injection demonstrates significantly reduced physical damage and thus improved embryo development with successful genome editing in up to 100% of living offspring. Hence, our novel approach for However, the impact of delivering these synthetic guide RNAs by electroporation on embryo viability has not been assessed side-by-side with conventional sgRNA. Thirdly, major results optimizing the electroporation conditions are based on experiments with zygotes from robust F1 hybrid mice whereas the vast majority of research questions demand transgenesis in the more sensitive but most commonly used inbred C57BL/6 mice in order to keep the results comparable and reproducible [To understand the mechanisms underlying human diseases biomedical research relies on the availability of animal models with precise recapitulation of human genetics. To this end, state-of-the-art genome editing by use of the CRISPR/Cas system in mouse zygotes has enabled the fast and scarless introduction of specific mutations with high efficiency \u20134. The moducible \u201315.Easy Electroporation of Zygotes), enables the introduction of specific mutations as efficient as delivery by conventional PNI while it significantly enhances embryo development in vitro and live birth rates in vivo. Genome editing accomplished in up to 100% of the founder mice demonstrates complete penetration of zygotes without any weakening of the zona pellucida. Thus, our EEZy approach represents an easily adaptable and broadly applicable technique for CRISPR-mediated mouse transgenesis which outperforms conventional PNI.To overcome these constraints, we combined the current data on electroporation in order to develop an easily adaptable protocol for efficient CRISPR/Cas9-mediated transgenesis of C57BL/6 zygotes with minimal technical demand. We aimed to omit any treatment weakening the zona pellucida and to establish the protocol on a standard electroporation device. Utilization of commercially available CRISPR/Cas9 components, most importantly synthetic guide RNAs, further simplified the process and enhanced the reproducibility as well as the embryo viability. We found that our improved CRISPR electroporation, which we refer to as EEZy and xylazine and euthanized by cervical dislocation. Carprofen was used as analgesic after surgery. All efforts were made to minimize suffering.All mouse protocols were in accordance with European, national and institutional guidelines and approved by the State Office of North Rhine-Westphalia, Department of Nature, Environment and Consumer Protection . Mice were kept in the specific and opportunistic pathogen free animal facility of the CECAD Research Center, University of Cologne, Germany, in individually ventilated cages at 22\u00b0C (\u00b1 2\u00b0C) and a humidity of 55% (\u00b1 5%) under 12 h light cycle with access to water and food TM crRNA) and generic tracrRNA were resuspended to 100 \u03bcM in sterile and nuclease-free T10E0.1 buffer ) prepared as described [in vitro transcription from the pSpCas9(BB)-2A-GFP (PX458) plasmid (Addgene #48138) from Feng Zhang and column purified . Guide RNAs were stored at -80\u00b0C. crRNA sequences are listed in Custom crRNA . Unless otherwise stated, mouse pre-implantation embryos were kept in pre-incubated M16 in a CO2 incubator or handled in M2 outside the CO2 incubator. M16 and M2 were prepared as described [in vitro fertilization without using reduced glutathione, essentially as described in published protocols with the minor modification of using Cook RVFE as fertilization medium and M16 for post fertilization wash steps instead of human tubal fluid media [Gt(ROSA)26Sor locus was a kind gift from Ralf K\u00fchn [2O ), incubated for 10 min at room temperature and add to 20 \u03bcM ssODN in 10 \u03bcl Opti-MEM. Each Cas9 RNP mix was kept on ice until use. For electroporation, zygotes were washed in batch through 5 drops of M2. After wash through one drop of Opti-MEM the zygotes were transferred with as little media as possible to 20 \u03bcl of Cas9 RNP mix. Using a P20 pipette, the entire solution was transferred to a pre-warmed 1 mm cuvette and placed in the BioRad Gene Pulser XCell electroporator. Two square wave pulses were applied . The zygotes were retrieved from the cuvette using a P100 pipette and two flushes of 100 \u03bcl M16 and transferred to a culture dish containing 500 \u03bcl M16. If indicated zygotes were treated with acidic Tyrode\u2019s solution for 10 s and the reaction stopped by adding M2. Zygotes for PNI were exclusively collected from C57BL/6NRj females upon natural mating as described above. After visual inspection for the presence of two pronuclei zygotes were placed in a paraffin oil covered M2 containing injection chamber. Microinjection was performed using an Axio Observer.D1 microscope (Zeiss) and microinjector devices CellTram and FemtoJet with TransferMan NK2 micromanipulators (Eppendorf). The CRISPR/Cas9 solution was injected into the male pronucleus using injection capillaries . If not stated otherwise the CRISPR/Cas9 injection mix was prepared as described for electroporation but containing 400 nM gRNA, 200nM Cas9 protein, 30 ng/\u03bcl Cas9 mRNA and 500 nM ssODN. Microinjected zygotes were transferred into 500 \u03bcl M16 and lysed embryos were removed approximately 1 h after injection. On the next day, 2-cell stage embryos were transferred unilateral into oviducts of pseudo-pregnant 0.5 dpc RjHan:NMRI females or continued to culture until the blastocyst stage at 3.5 dpc.A step-by-step protocol for EEZy is available on protocols.io in a thermocycler at 65\u00b0C for 15 min followed by 95\u00b0C for 15 min and stored at -80\u00b0C until use. Except depicted otherwise, two sequential rounds of PCR with Herculase II fusion DNA polymerase were performed according to the manufacturer\u2019s instruction using 2 \u03bcl DNA template. The primer sequences and thermocycling conditions are listed in p-value of 0.05. The collection of all data used for statistical analysis can be found in For calculation of statistical significance, standard deviation and arithmetic mean of differences Prism (GraphPad) and Excel (Microsoft) were employed. Box plots were generated by Prism. Statistical significance was assessed using a two-tailed, unpaired Student\u2019s T-test. Differences were considered significant below a In order to establish an easily adaptable protocol for CRISPR/Cas9 electroporation in C57BL/6N zygotes we intentionally utilized a standard electroporator (BioRad Gene Pulser XCell). Similar devices are broadly available in biomedical laboratories. Two pulses of 3 ms at 30 V were previously demonstrated as the ideal balance between editing efficiency and embryo survival for electroporation with this device . We chosNphs2 gene by electroporation of solely commercially available CRISPR/Cas9 components and analyzed the genotype of the blastocysts by RFLP analysis. In a first set of experiments, we determined whether previously suggested additives to the electroporation buffer are necessary to maintain editing efficiency and embryo development [Atp1a1) using our EEZy approach . The use of these commercially available pgRNAs for electroporation has recently been described but their impact on editing efficiency and embryo development compared to conventional sgRNA is unknown [Nphs2 26Sor locus and a plasmid vector already successfully used by conventional PNI [We also assessed whether our EEZy approach is effective for delivery of plasmid targeting vectors to generate CRISPR-mediated large genomic integrations as such has not been published to date using electroporation. We attempted to integrate a Venus reporter transgene by electroporation of Cas9 RNPs targeting the onal PNI . DespiteTmem218) employing our EEZy approach in mice of the C57BL/6J background nor EEZy of CRISPR components significantly affected embryo development. Next, we directly compared the results of our EEZy approach to transgenesis by conventional PNI using the same knock-in strategy for Nphs2 as before. In line with previously published results, on average 82% of the zygotes survive in our PNI Click here for additional data file.S2 FigAtp1a1 gene by electroporation. PCR genotyping and the percentage of transgenic 3.5 dpc embryos of zygotes pre-treated for 10 s with acidic Tyrode\u2019s solution (weakened) or untreated (intact) is depicted. Arrows indicate the expected size of the band for the PCR with primers flanking the endogenous locus (WT locus) or amplifying the myc tag sequence (transgene). PCR controls from untreated embryos (WT) and without DNA template (H2O) are included. (B) Developed blastocysts upon treatment with acidic Tyrode\u2019s solution for 10 s. (C) Quantification of Sanger sequencing of Tmem218 targeted blastocysts generated by EEZy. (D) Quantification of Sanger sequencing of biopsies from Lyn transgenic mice generated by EEZy using C57BL/6J zygotes. HDR = solely the desired mutation, HDR + INDELs = mixture of the desired mutation and INDELs, INDELs = mono- and biallelic INDELs, WT = no genome editing. N = Number of embryos/animals.(A) Myc tag insertion into the (PDF)Click here for additional data file.S3 FigGt(ROSA)26Sor targeting vector (6 kB) harboring a Venus reporter transgene fused to a splice acceptor site (Gt(ROSA)26Sor SA-Venus) was electroporated into zygotes together with a Cas9 RNP targeting the Gt(ROSA)26Sor locus. PCR genotyping for integration of the transgene and amplification of the WT locus are depicted. PCR controls from untreated embryos (WT), without DNA template (H2O) and Gt(ROSA)26Sor SA-Venus targeted blastocysts (+transgene) are included and the expected size for amplification of the transgene (tg) depicted. Evaluation of NHEJ at the endogenous Gt(ROSA)26Sor locus is shown by partial resistance to XbaI digest (mut). Controls from PCR amplicons of WT blastocysts with (WT+XbaI) and without XbaI (WT-XbaI) are included.A circular (PDF)Click here for additional data file.S4 FigNphs2-targeted blastocysts upon PNI. Results of four independent experiments (#1\u20134) with the percentage of transgenic blastocysts are depicted (n = 4). PCR controls from untreated blastocysts (WT) and without DNA template (H2O) and RFLP controls from untreated blastocysts (WT) are depicted. The data are quantified in RFLP analysis of (PDF)Click here for additional data file.S1 Table(PDF)Click here for additional data file.S2 Table(PDF)Click here for additional data file."} +{"text": "BRCA1/2 mutation ovarian cancer. However, antitumor responses are observed in only approximately 40% of patients and the impact of baseline clinical factors on response to treatment remains unclear. Although platinum sensitivity has been suggested as a marker of response to PARP inhibitors, patients with platinum-resistant disease still respond to olaparib.The PARP inhibitor olaparib was recently granted Food and Drug Administration (FDA) accelerated approval in patients with advanced BRCA1/2 mutation ovarian cancers were included. The interval between the end of the most recent platinum chemotherapy and PARPi (PTPI) was used to predict response to olaparib independent of conventional definition of platinum sensitivity. RECIST complete response (CR) and partial response (PR) rates were 35% in patients with platinum-sensitive versus 13% in platinum-resistant (p<0.005). Independent of platinum sensitivity status, the RECIST CR/PR rates were 42% in patients with PTPI greater than 52 weeks and 18% in patients with PTPI less than 52 weeks (p=0.016). No association was found between baseline clinical factors such as FIGO staging, debulking surgery, BRCA1 versus BRCA2 mutations, prior history of breast cancer and prior chemotherapy for breast cancer, and the response to olaparib.108 patients with advanced BRCA1/2 mutation ovarian cancers from eight different cancer centers and their antitumor response to olaparib.We conducted an international multicenter retrospective study to investigate the association between baseline clinical characteristics of patients with advanced PTPI may be used to refine the prediction of response to PARP inhibition based on the conventional categorization of platinum sensitivity. BRCA1 and BRCA2 (BRCA1/2) genes has greatly enhanced our knowledge of the DNA repair pathways involved in cancer progression and has led to the exploitation of the concept of synthetic lethality in cancer therapy [BRCA1/2 mutation cancers. During this time, multiple clinical trials have investigated the clinical activity of different PARP inhibitors in a range of cancers. Olaparib is the most extensively studied PARP inhibitor in patients with advanced BRCA1/2 mutated ovarian cancer and has now received Food and Drug administration (FDA) and European Medicines Agency (EMA) regulatory approval in the relapsed and maintenance treatment settings, respectively.The identification of BRCA1/2 mutation ovarian cancer has been assessed in a number of phase I/II trials. Overall, Response Evaluation Criteria In Solid Tumors (RECIST) complete or partial responses (CR/PR) or Gynecologic Cancer InterGroup (GCIG) CA125 antitumor responses ranged between 33%-59%, with a clinical benefit rate (RECIST or GCIG CA125 responses and RECIST stable disease) of 46-52% [BRCA1/2 mutated ovarian cancer (NCT02282020).The clinical activity of olaparib in patients with advanced germline f 46-52% . The antBRCA1/2 mutation ovarian cancer, not all patients achieve the same level of benefit to olaparib. This highlights the need to identify individual patient predictive biomarkers of response and resistance to guide treatment decisions. A number of translational clinical studies have now been initiated to identify molecular biomarkers of response to olaparib and other PARP inhibitors. Several studies are investigating the different molecular markers that might predict for antitumor response to olaparib and other PARP inhibitors beyond germline BRCA1/2 mutations, including somatic BRCA1/2 mutations, functional classifiers of homologous recombination deficiency, gene expression profiling and genomic scarring signatures [While phase I/II clinical trials with olaparib have reported impressive objective response rates in patients with advanced gnatures \u201312.BRCA1/2 mutation ovarian cancer, including the effects of platinum sensitivity as potential factors that may be used in the prediction of patients benefit to olaparib. In addition, we assessed the association between antitumor response to olaparib treatment and progression-free survival (PFS) and overall survival (OS).Earlier observations from limited clinical data suggested a potential relationship between prior sensitivity to platinum-based chemotherapy and antitumor responses to olaparib . This haBRCA1/2 mutation advanced ovarian cancer from eight cancer centers who had been treated with olaparib at a dose of 200mg BID or greater were included in our study patients had germline BRCA2 mutations. Forty (38%) patients had a past history of breast cancer, of whom 20 (22.2%) received one previous line of chemotherapy for breast cancer.A total of 108 patients with germline BRCA1 (27.6%) mutations versus those with BRCA2 (27%) mutations (p=0.31). There was no association between having a past history of breast cancer or receiving prior chemotherapy for breast cancer, and achieving RECIST antitumor responses to olaparib (Table Patients who only had one prior line of chemotherapy prior to receiving olaparib had significantly higher rates of RECIST CR/PR compared to those who received more than one prior line of treatment (p=0.005). No differences in RECIST CR/PR rates were noted between patients with germline BRCA1 7.6% mutaThe median PFS was 70 weeks for patients who had achieved RECIST CR/PR with olaparib therapy, compared to 28 weeks for patients without RECIST CR/PR Figure . Median At the time of enrolment to each trial, 64% of patients were platinum sensitive, while 36% were platinum resistant. Higher likelihood of RECIST CR/PR rates were observed in patients with platinum-sensitive ovarian cancer (35%), in contrast to those with platinum-resistant disease (13%) (p=0.02). Although the RECIST CR/PR rate was significantly lower in platinum-resistant patients compared to those with platinum-sensitive disease , 5 of 38 (13%) patients conventionally defined as platinum-resistant still achieved RECIST CR/PR, suggesting that platinum response status is not an absolute predictor of response to olaparib.The median PTPI was 53 weeks (range 4-244 weeks). Median PTPI was significantly longer for patients with platinum-sensitive ovarian cancer (68.7 weeks) compared to patients with platinum-resistant disease (25.9 weeks) (p<0.0001). Independent of their platinum response status, patients with PTPI of greater than 52 weeks had higher CR/PR rates than those with PTPI of less than 52 weeks .In order to study the association of PTPI with response, we compared the RECIST response rate to olaparib in patients who were classed as platinum-sensitive before commencing olaparib and had PTPI of greater than 24 weeks (\u2018best-responders\u2019), to patients who were platinum-resistant and had PTPI of less than 24 weeks (\u2018least-responders\u2019). Fifty-eight patients were defined as \u2018best responders\u2019 and 20 patients as \u2018least responders\u2019. The RECIST CR/PR rates achieved with olaparib with the \u2018best responders\u2019 group were significantly higher than the \u2018least responders\u2019 group . We did not identify any association between response to olaparib and other clinical baseline characteristics, including initial FIGO staging or cytoreductive surgery.BRCA1/2 mutation ovarian cancer.In this international multicenter retrospective study, we investigated the baseline clinical characteristics that may predict for antitumor responses to treatment with the PARP inhibitor olaparib in patients with advanced BRCA1/2 mutations have homologous recombination deficient tumors, which may be treated effectively with PARP inhibitors [BRCA1/2 mutation tumors, including those with ovarian cancer [BRCA1/2 mutation ovarian cancer treated with olaparib in clinical trials. These trials confirmed the impressive activity of olaparib in patients with recurrent BRCA1/2 mutant ovarian cancers, even in a proportion of patients with platinum-resistant disease [The hypothesis underlying the concept of synthetic lethality is that patients harboring deleterious germline n cancer , 13, 14. disease . A recen disease .A number of prospective and retrospective studies are currently ongoing to identify molecular biomarkers that may predict antitumor response to olaparib and other PARP inhibitors \u201312, 16. BRCA1/2 mutation ovarian cancer. Our data suggest that patients with a longer platinum-free interval, even after becoming resistant to platinum-based chemotherapies, are more likely to respond to olaparib and potentially other PARP inhibitors. This is consistent with some studies, which have suggested that a longer platinum-free interval may increase antitumor response rates to subsequent treatment, including rechallenging patients with platinum-based therapy [BRCA1/2 mutation ovarian cancers.PTPI may be a useful indicator for informing both timing and sequence of non-platinum containing treatment regimens during the treatment journey of patients with advanced therapy , 18. It BRCA1 versus BRCA2 mutations [BRCA1 mutations and 30 with BRCA2 mutations, no differences in antitumor response was observed between both groups of patients. Details on the specific BRCA1/2 gene mutations and their association with response to olaparib were not collected. This requires a larger series of patients that may further explain the lack of antitumor responses to PARP inhibitors in patients with advanced BRCA1/2 mutation ovarian cancer.It has previously been speculated that the antitumor response to olaparib may be different in patients who have germline utations . In thisBRCA1/2 mutation ovarian cancer treated with the PARP inhibitor olaparib, a limitation of this study was the involvement of a relatively small series of patients that prevented further statistical analysis such as regression analysis. Future studies with a larger data set will enable linear analysis to be undertaken to identify a more precise PTPI fit model to predict antitumor response to PARP inhibitors. The patients in our study have mostly undergone several lines of chemotherapy, so it is still possible that there is a population of either BRCA1 or BRCA2 patients who are excellent responders to first line chemotherapy and don't need olaparib or alternatively some who are very platinum resistant and don't survive to get olaparib.Despite this being one of the largest retrospective series of clinical data from patients with advanced BRCA1/2 mutation ovarian cancer heralded a new era in precision medicine in patients with advanced ovarian cancer. Several other PARP inhibitors are currently at different stages of clinical development in patients with BRCA1/2 mutation ovarian cancer. Despite this, improved strategies for selecting patients who may benefit most are urgently required, including clinical factors that can be implemented easily without additional delays, costs or logistical issues. We have shown that PTPI may potentially be an important clinical biomarker that should be considered when treating patients with advanced BRCA1/2 mutation ovarian cancer with a PARP inhibitor, and warrants further investigation either in conjunction with or independent of conventional platinum response status.The accelerated approval of olaparib by the FDA and EMA in patients with advanced BRCA1/2 mutation ovarian cancer from 8 cancer centers , which had participated in clinical trials of olaparib monotherapy between April 2006 and August 2014. This study was approved by the Royal Marsden Clinical Research and Development Committee.Detailed clinical data were collected retrospectively from patient records of 108 patients with advanced recurrent germline Only patients who had been treated with olaparib at doses of 200 mg tablet or capsule formulation or more twice daily in the relapsed setting were entered to this study. This minimum dose of 200 mg BID was selected on the basis of pharmacodynamic and antitumor activity of olaparib from previous studies, where doses of 200 mg BID resulted in significant target inhibition and antitumor responses . This stBRCA1/2 status, prior chemotherapy, platinum-based chemotherapy sensitivity status at the time of enrolment, prior history of breast cancer or treatments received for breast cancer, RECIST and GCIG CA125 responses, survival, response rates and the interval between last platinum-based chemotherapy to the start of olaparib treatment.Baseline patient characteristics were collected and entered into a standardized database; these included details on demographics, surgical debulking, Patients were defined as platinum-sensitive or platinum-resistant when the platinum-free interval was more than 12 months or less than 6 months, respectively . Patient2, odds ratios (OR) and Fisher's exact probability tests were used for statistical analyses. Survival curves were generated using GraphPad Prism 6.Pearson Chi"} +{"text": "Scientific Reports6: Article number: 3420710.1038/srep34207; published online: 09262016; updated: 12092016This Article contains an error in the legend of Figure 2.\u201cMean activity % and standard error of migrant (blue) and resident (orange) European blackbirds (Turdus merula) in 30\u2009minute intervals four days prior to departure\u201d.should read:\u201cMean activity % and standard error of migrant (blue) and resident (orange) European blackbirds (Turdus merula) in 30\u2009minute intervals seven days prior to departure\u201d."} +{"text": "CsEXPA1) on the global gene expression pattern during early and late phases of etiolated hypocotyl growth.Expansin increases cell wall extensibility to allow cell wall loosening and cell expansion even in the absence of hydrolytic activity. Previous studies showed that excessive overexpression of expansin gene resulted in defective growth and altered cell wall chemical composition [3]. However, the molecular mechanism on how the overexpression of non-enzymatic cell wall protein expansin can result in widespread effects on plant cell wall and organ growth remains unclear. We acquired transcriptomic data on previously reported transgenic Arabidopsis line [1] to investigate the effects of overexpressing a heterologus cucumber expansin gene ( Specification TableDirect link to deposited datahttp://www.ncbi.nlm.nih.gov/sra/SRP076440.Value of the data\u2022This is the first transcriptome data of etiolated Arabidopsis hypocotyl overexpressing an expansin gene compared with previous reports CsEXPA1 in tomato resulted in shorter plants The overexpression of \u2022The present dataset is valuable for the identification of the genes which respond to expansin gene overexpression during the early and late phases of hypocotyl growth.\u2022This information will be useful for investigating global gene expression during hypocotyl growth, and for studying the effect of overexpressing a cell wall loosening factor on other cell wall related genes or regulatory factors to identify possible feedback mechanism responsible for growth defect.1pOpON::CsEXPA1 Arabidopsis hypocotyls harvested on day 3 and day 5; each set with three biological replicates. This transcriptomic dataset was generated by QuantSeq. 3\u2032 mRNA sequencing SRP076440 (http://www.ncbi.nlm.nih.gov/sra/SRP076440) under the BioSample accession number SAMN05192734.Data reported here describe the sequencing results obtained22.1pOpON::CsEXPA1CsEXPA1). Seed sowing, growing media and conditions followed as previously described This study utilised previously reported transgenic Arabidopsis line 2.2RNA from pools of 100 hypocotyls was extracted using TRIzol (Invitrogen) according to manufacturer's instruction. RNA purity and integrity was measured using the ND-1000 Nanodrop spectrophotometer and Agilent 2100 bioanalyzer , respectively. RNA samples were cleaned using DNAse I kit according to the Rapid out removal DNA kit instruction (Thermoscientific) and converted into cDNA by using QuantSeq. 3\u2032 mRNA-Seq Reverse (REV) Library Prep Kit (Lexogen) 2.3https://www.lexogen.com/quantseq-data-analysis/). To quantify transcript abundance, the processed reads (FASTA) were mapped to Arabidopsis genome reference . The mapping was performed using bowtie2 CsEXPA1 gene from cucumber. Data from this study can be compared with previously reported expression data from the suppression of endogenous expansin genes Raw sequencing reads (FASTQ) were processed individually to checked for per base sequence quality and screened for the presence of any Illumina adaptor/ overrepresented sequences and cross-species contamination through the AGRF quality control (QC) pipeline as per Lexogen QuantSeq data analysis workflow ("} +{"text": "Equus caballus solute carrier family 9, subfamily A2 was one of the four differentially expressed transcripts linked to osteoclast activity. Osteoblasts were hyperplastic and hypertrophic in bone marrow from affected horses. Biological pathways associated with skeletal morphogenesis were significantly enriched in affected horses. The 30 differentially expressed genes in affected lymph nodes were associated with inflammatory responses. Evidence of infectious agents was not found. The SAO affected bone marrow molecular signature demonstrated increased transcription and heightened activation of osteoblasts. Increased osteoblastic activity could be part of the pathological mechanism for osteoporosis or a compensatory response to the accelerated osteolysis. Transcriptome data offer gene targets for inquiries into the role of osteocytes and osteoblasts in SAO pathogenesis. Viral or bacterial infectious etiology in SAO is less likely based on metatranscriptomic and metagenomic data but cannot be completely ruled out.Osteoporosis has been associated with pulmonary silicosis in California horses exposed to soils rich in cytotoxic silica dioxide crystals, a syndrome termed silicate associated osteoporosis (SAO). The causal mechanism for the development of osteoporosis is unknown. Osteoporotic lesions are primarily located in bone marrow-rich sites such as ribs, scapula and pelvis. Gene transcription patterns within bone marrow and pulmonary lymph nodes of affected horses may offer clues to disease pathobiology. Bone marrow core and tracheobronchial lymph node tissue samples harvested postmortem from affected and unaffected horses were examined histologically and subjected to RNA sequencing (RNA-seq). Sequenced data were analyzed for differential gene expression and gene ontology. Metatranscriptomic and metagenomic assays evaluated samples for infectious agents. Thirteen of 17 differentially expressed transcripts in bone marrow were linked to bone and cartilage formation such as integrin binding bone sialoprotein (log The settings included statistical enrichment test defaults and Equus Caballus (EquCab2.0) as the reference species. Statistical significance was set as p-value below the Bonferroni correction was used to accounted for multiple comparisons in the analysis. PANTHER Pathways and GO-Slim Biological Processes were selected as the Annotation Data Sets for analysis.The files of genes from the DGE analysis with ENSEMBL gene IDs and fold-change expressed in logEquus Caballus taxonomy labels assigned by KrakEN.[Because the pathogenesis of SAO is unknown, non-equine reads were evaluated for similarity to sequences of infectious agents that could be attributed to viruses or bacteria. Trimmed reads were evaluated with KrakEN, a k-mer based metagenomic classification tool, using a combined horse, viral, and bacterial database, followed by the BRAKEN metagenomic classification pipeline for transcription frequency of non-equine genetic material, and reported as fractions relative to the y KrakEN.\u201325+ tissues for viral transcripts. Samples of SAO+ BM and tLN SAO1, SAO7, SAO3, SAO9 (not used in transcriptome study)), as well as buffy coat and lung tissues were pooled. Viral capsid-protected nucleic acid was extracted and enriched according to an established protocol.[Metagenomic analysis evaluated SAOprotocol. An IllumSufficient depth of sequencing and an adequate mapping rate were achieved for the DGE analysis. The average reads across BM and tLN samples were ~25.7 million reads/sample. Following trimming adapter contamination and removal of low quality reads and PCR duplicates with expHTS software, the average mapping rate to EquCab2.0 was 92.63%, where 54.6% reads uniquely aligned to an annotated horse gene and 32.4% aligned to regions of genome without annotation. 5.28% of reads aligned to multiple regions in the genome and 0.35% aligned with some annotation overlap, therefore, these reads were not assigned to a specific gene.+ and SAO- transcriptomes in BM and tLN tissues were most influenced by the tissue origin . Among the genes with increased transcripts, Equus caballus solute carrier family 9, subfamily A2 (SLC9A2) expression is linked to osteoclast differentiation and survival.[TRPV4).[OMD) [DLX3)[DAPK2),[+ horses. Five of the differentially expressed genes associated with the developing skeleton include DLX3,[SP7),[COL24A1),[COLXIA2),[CHAD). [BSP),[OC),[SMPD3).[Overall 12 of 17 differentially expressed genes in SAOormation . Sixteensurvival. Transien.[TRPV4). Osteomod4).[OMD) and distD) [DLX3) gene pro)[DAPK2),, 52 was ude DLX3, Sp7 tranX3,[SP7),, 41 collCOL24A1), collagenCOLXIA2), and chon,[CHAD). Increase). [BSP),, 61 osteSP),[OC), and sphi,[SMPD3)., 46+ horses were included for their functional relevance to the silicosis induced inflammatory process , increased coagulation , and decreased signaling through PDGF were over-represented pathways in affected tLN samples.Increased signaling through the integrin , PDGF and cadherin pathways were found in the SAOBM and tLN sampled from the same site as tissues harvested for RNA were evaluated microscopically to assess the biological relevance of the transcriptome data.+ and unaffected BM samples. Mean (\u00b1 SD) ratio of bone forming surface to total bone surface perimeter was greater (p<0.001) in SAO+ horses (0.31 \u00b1 0.14) than unaffected horses (0.05 \u00b1 0.04). Osteoblast hypertrophy, captured as apical to basilar height, was significantly greater (p = 0.002) in SAO+ horses (11.6 \u00b1 2.2 \u03bcm) than unaffected horses (5.4 \u00b1 3.8 \u03bcm). Mean ratio of bone resorption surface to total bone surface perimeter was greater (p = 0.016) in SAO+ horses (0.15 \u00b1 0.12) than unaffected horses (0.03 \u00b1 0.04). However, few osteoclasts were present in sections (ranging from 0 to 6 per image area) compared to the other cell populations and numbers per total bone perimeter were not statistically different (p = 0.401) between affected and unaffected horses. However, osteoclasts were subjectively more ubiquitous in SAO+ horses with greater cell volume and nuclei numbers and Alteromonas mediterranea . One SAO+ BM sample had Mycobacterium tuberculosis as the second most abundant species (0.00017% of reads) without microscopic evidence of granulomatous lesions.To determine if the presence of infectious agents was associated with the differential transcriptome data from SAO+ horses to determine if viral genetic material could be detected. No viral RNA, despite the findings of granulovirus in the metatranscriptomic analysis, was identified in pooled tissues .Metagenomic analysis was conducted on selected tissues from 3 SAO+ BM transcriptome relative to unaffected animals. DGE and GO enrichment analyses demonstrated pro-inflammatory activity in SAO+ tLNs, but inherent tissue heterogeneity may have diluted a more distinct transcriptome pattern. There was no evidence of infectious co-factors in SAO etiology. This study provides a novel viewpoint of a dominant molecular signature of bone formation (increased osteoblastic activity) in a condition that has been morphologically defined by osteoclastic hyperactivity and clinically significant osteoporosis.[The cardinal lesions of SAO in tLN and bone marrow-rich skeletal sites were present in all affected horses, supporting circumstantial evidence for their interconnected role in the pathobiology of SAO. The moleoporosis. This dat+ BM tissue suggests a number of pathogenic possibilities. Lack of anticipated transcriptome pattern associated with increased signaling through pro-osteoclastogenic RANK may reflect the low frequency of these transcripts within the complex cellular milieu of BM. Alternatively, the lack of evidence for a dominant osteoclastogenic signal may reflect the brief temporal nature of such a signal. Regardless, within lesions characterized by abundant bone removal, histologic evidence of overt osteoblastic activity is concurrently present. By design, tissue RNA samples were obtained from grossly normal bone, anatomically distant from osteoporotic lesions.Although unexpected, the paucity of transcripts associated with osteoclast differentiation in SAO+ BM transcriptome revealed predilection for skeletal formation transcription. This finding was supported by morphologically increased osteoblast activity in affected BM samples compared to quiescent bone surfaces of unaffected tissues. Osterix (Sp7) is one of the differentially increased transcripts indicative of enhanced osteoblast differentiation.[Sp7 like IBSP, BGLAP and COL24A1 provided further transcriptional evidence of increased osteoblast differentiation in SAO+ BM.[DGE and GO enrichment analyses of the SAOntiation. Expressi SAO+ BM., 63, 64DLX3 and CDH15 shown to have a higher transcription level than osteoblasts in one study.[Marked osteoblastic activity coupled with hyperactive osteoclasts is characteristic of SAO lesions and other bone diseases like fibrous osteodystrophy. However, overrepresented activation of osteoblasts away from regions of intense osteolysis was not anticipated. Activation of osteoblasts as a systemic reparative or compensatory mechanism despite minimal osteoclastic activity within tested sample is a potential explanation. The results could be considered in the context of increased mechanical loading distributed along weakened bone and increased differentiation of BM stromal cells toward osteo-chondral lineage. The indune study.+ BM. Increased transcripts in our data are downstream of the master regulators of skeletal formation like bone morphogenetic protein (BMP) [COLXIA2, SMPD3, TRPV4, CHAD, and ANO5 are associated with cartilage tissue functions [Mechanisms involved in fetal bone formation are rekindled during endochondral ossification in skeletal fracture repair and potentially activated in SAOSalazar) and runtSalazar) , Transcriunctions , 47 alth+ BM was speculatively linked to the increased transcription of SLC9A2, DLX3, TRPV4 and OMD.[Increased osteoclast differentiation and activity in SAO and OMD., 30, 36 and OMD. Results + horses, but the potential significance has not been realized.[Our results suggest a multifocal rather than systemic activation of an osteolytic trigger in SAO. The multifocal distribution of osteolysis has been noted in post mortem examination and on nuclear scintigraphic examination of SAOrealized., 3 The pHSD17B6, in SAO+ BM is unclear because function of the equine isoenzyme is not defined in the literature. HSD17B6 might have an important role in bone as some isoenzymes catalyze the interconversion of estrone (E1) to \u201costeoprotective\u201d estradiol E2).[HSD17B6 has been identified as a mediator of the conversion of androgen substrates into inactive forms.[HSD17B6 in SAO+ horses could be a response to accelerated bone loss, or part of the disease pathobiology resulting in excessive osteolysis.The role of the only down-regulated transcript, diol E2). In humanve forms. DependinCompared to GO and DGE analysis, PANTHER pathway evaluation of the BM transcriptome did not explicitly demonstrate osteoblast-associated functions. Pathway analysis showed increased signaling through the integrin and cadherin pathways broadly used by cells in the BM tissue.+ horses. DGE analysis did not reveal a transcription pattern to explain the association between the skeleton and lung conditions in SAO. The 30 functionally defined differentially expressed genes within the SAO+ tLNs showed variably regulated transcripts broadly related to immune system functions. Theoretically, connections between silicosis and osteoporosis could be made with some transcripts. For example, an increase in DMP1 transcription echoes BM tropism of an osteolytic trigger in SAO because it was up-regulated in bone tropic metastatic lung cancers.[DMP1 in non-mineralized tissues may indicate a wider purpose for extra-skeletal regulation of mineralization.[PTX3 up-regulation may have some connection with silicosis as a promoter in TNF-\u03b1 and IL-1\u03b2 production through binding NF-\u03baB elements, as demonstrated in vitro.[\u03b1 has been implicated in the pathobiology of silicosis.[PTX3 is increased in cultured human bronchial epithelial cells exposed to cytotoxic silicates.[Both DGE and GO enrichment analyses showed evidence for nonspecific inflammation in the tLNs of SAO cancers. Physiololization. PTX3 up-in vitro. Increaseilicosis., 73 Inteilicates. Inherent+ horses with advanced osteoporosis resulted in challenging barriers for patient recruitment. Isolation of the specific cell targets from BM and blood of live animals, such as osteoblasts or osteoclasts using laser dissection, and monocytes with a cell sorting technique may prove useful for future studies. The implication of the results from the metatranscriptomic analysis in our study was unclear since the low frequency transcripts of an arthropod virus and marine bacteria were present in all the samples. Misclassification of the non-equine transcripts was considered as a possibility. The finding of low frequency transcripts of Mycobacterium tuberculosis in one SAO+ BM sample was not supported by clinical and pathological examination. Elimination of the poly A RNA selection step would potentially resolve limitations of the metatranscriptomic analysis in including infectious agents that do not transcribe their genetic material in this RNA form.Modification of the tissue target, sample collection, and RNA preparation protocols, as well as identification of SAO clinical markers would curtail some of the limitations for the prospective investigation of this condition. Because necropsy and histopathology remain the gold standard diagnostic tools to confirm SAO, only horses in terminal condition were recruited for the study. Concerns over transportation of severely affected SAOIn conclusion, the data revealed an SAO associated transcriptome profile in BM tissue skewed toward skeletal formation which is also supported by microscopic findings (at bone sites distinct from lesions of osteolysis). The hypothesized molecular signal of a pathogenic link connecting the pulmonary and skeletal lesions was not elucidated by our results, and increased transcripts associated with osteoclast differentiation and activation were relatively fewer than osteoblast related transcripts. A clear transcriptome pattern was not identified between affected and unaffected animals due to the cellular heterogeneity in the tLN samples. Specific cell targets from the BM and tLN tissues would address the potential dilution of the disease-specific transcriptome pattern. Transcriptome profile of osteoclasts, osteoclast precursors or macrophages from osteolytic SAO lesions may provide better understanding of osteoclast dysregulation. The effect of ongoing pulmonary inflammation and silicosis on osteoblasts and regulation of bone formation warrants investigation in light of our findings. This is the first study that uses RNA-seq technology in investigation of disease mechanism in the equine co-morbid condition though RNA-seq has been used in research of other equine diseases.\u201379 SAO mS1 FigThe sequenced samples transcript patterns cluster based on tissue in this MDS plot that simultaneously evaluated BM and tLN transcriptomes together.(TIF)Click here for additional data file.S2 FigBM MDS plot demonstrates clustering of cases (red) based on bone phenotype. The group with mild osteoporosis (encircled in black) co-localized with control cases.(TIF)Click here for additional data file.S3 FigtLN MDS plot demonstrates clustering of control cases and wider dispersal of the cases transcriptome profiles.(TIF)Click here for additional data file.S4 Fig+ tLN with raw p-value <0.05 are included in the bar chart. Bars represent -Log 10 converted p values. The most enriched biological processes related infectious agents like virus and bacterial elements in the SAO(TIF)Click here for additional data file.S5 Fig(TIF)Click here for additional data file.S6 Fig(TIF)Click here for additional data file.S1 Table+ and Control horses.Animal signalment and quality of RNA extracted from BM (BM RIN) and tLN (LN RIN) for SAO(DOCX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S1 Protocol(DOCX)Click here for additional data file."} +{"text": "We described the treatment rationale and procedure for a phase II study of docetaxel plus ramucirumab for non\u2013small cell lung cancer (NSCLC) patients with brain metastasis . Combination therapy of angiogenetic inhibitor with chemotherapy improved the outcome of patients with brain metastasis in previous reports; however, the efficacy of ramucirumab, a vascular endothelial growth factor receptor-2 monoclonal antibody, for brain metastasis has not been shown.2) and ramucirumab (10\u200amg/kg) every 21 days until disease progression. The primary endpoint is progression-free survival (PFS), and secondary endpoints are overall survival, intracranial PFS, response rate, and safety. Sixty-five participants will be recruited from September 2017 to December 2019 and followed up for 1 year after final registration. The results from this study may suggest a treatment option for brain metastasis in NSCLC.This RAMNITA study is a prospective, multicenter, open-label, single-arm phase II study designed to evaluate efficacy and safety of docetaxel and ramucirumab for advanced NSCLC patients with brain metastasis. Eligible patients will receive docetaxel (60\u200amg/mThe protocol was approved by the institutional review board of each study center. Written informed consent will be obtained from all patients before registration, in accordance with the Declaration of Helsinki. Although efficacy of chemotherapy for brain metastasis is limited, radiological therapies, including stereotactic radiosurgery (SRS) and whole brain radiotherapy, or surgical resection may be used for local control of brain metastasis.Non\u2013small cell lung cancer (NSCLC) is usually diagnosed at an advanced stage of the disease, and brain metastasis is a common complication in NSCLC patients, with >10% of patients with NSCLC presenting with brain metastasis at their first hospital visit In cancer, tumor angiogenesis owing to overexpression of angiogenetic factors, such as vascular endothelial growth factor (VEGF) receptor, create an abnormal tumor microenvironment characterized by hypoxia and acidosis, and interstitial hypertension owing to vascular hyperpermeability, which reduces drug penetration into tumors. Antiangiogenetic agents can decrease tumor vascular permeability and interstitial fluid pressure by inhibiting of tumor angiogenesis, and thereby improve the efficacy of coadministered anticancer drug(s). Furthermore, bevacizumab combined with cytotoxic agents improved the survival of patients with newly detected brain lesions.,7Previous research revealed angiogenesis via the VEGF pathway is involved in the formation of brain metastasis. Subset analysis of AVAiL trial data showed that bevacizumab combined with platinum-doublet chemotherapy significantly decreased brain metastasis development. The REVEL study was a global, randomized, placebo-controlled, double-blind, multicenter phase III study comparing docetaxel plus ramucirumab combination treatment with docetaxel treatment (docetaxel plus placebo) in patients with stage IV NSCLC who showed disease progression after platinum-based therapy. This study showed that second-line docetaxel plus ramucirumab combination treatment of patients with stage IV NSCLC improves progression-free survival (PFS), overall survival (OS), and response rate; however, the efficacy of ramucirumab for brain metastasis remained unclear.,9 The current trial is designed to evaluate the efficacy and toxicity of a docetaxel plus ramucirumab regimen as a treatment for NSCLC with brain metastasis.Ramucirumab is a human recombinant IgG1 monoclonal antibody that specifically binds to the extracellular domain of VEGF receptor-2 with high affinity, preventing the binding of VEGF ligands and receptor activation.22.1The RAMNITA study is an open-label, single-arm trial of NSCLC with brain metastasis. Figure 2.22 on day 1 of a 3-week cycle will be continued until disease progression or fulfillment of the criteria of treatment cessation. No dosage adjustment according to age, body weight, sex, ethnicity, and smoking status is warranted.Intravenous administration of ramucirumab 10\u200amg/kg plus docetaxel 60\u200amg/mThis study has been conducted in compliance with the principles of the Declaration of Helsinki and registered in the University Hospital Medical Information Network database (UMIN000024551).2.3Key eligible criteria are listed below.(1)Histological or cytological confirmation as advanced NSCLC(2)Asymptomatic brain metastasis is present. SRS (including Gamma Knife or Cyber Knife) before enrollment is allowed.(3)A history of chemotherapy(4)Performance status (ECOG) is 0 or 1.(5)Age \u226520 years(6)Bone marrow, coagulation factor activity, and organ functions have all been confirmed as normal.(7)Life expectancy of at least 3 months(8)If sexually active, patients must be postmenopausal, surgically sterile, or using effective contraception.(9)Other cancers must have been treated curatively and followed without evidence of recurrence for at least 3 years before acquisition of consent.(10)Provision of written informed consent(1)Receiving other anticancer therapy currently(2)Presenting symptoms caused by brain metastasis(3)History of whole brain radiotherapy or surgical resection for brain metastasis(4)Presence of meningeal metastasis(5)Bleeding into the brain metastasis or other central nervous system hemorrhage within 21 days before acquisition of consent(6)Intratumor cavitation or major blood vessel invasion or encasement by cancer(7)Arterial thromboembolic events, venous thromboembolism events, or serious bleeding complications within 6 months before acquisition of consent(8)Other serious complications(9)Patients who are pregnant, nursing, or possibly pregnant(10)Uncontrolled metabolic disorder, including diabetes mellitus(11)Active or uncontrolled clinically serious infection2.4After the eligibility criteria have been met and the patients have provided informed consent, eligible patients will be registered and the planned treatment will be initiated by the investigators. Accrual began in May 2017 and should continue for 3 years.2.5Patients will be administered docetaxel and ramucirumab every 3 weeks beginning on day 1 and continue until disease progression or the fulfillment of the discontinuation criteria. Tumor assessments by imaging should be performed as scheduled every 6 weeks (\u00b12 weeks) during 8 months following protocol treatment commencement, and every 9 weeks (\u00b12 weeks) thereafter, until progressive disease is documented on imaging.The antitumor effect is evaluated according to the \u201cNew response evaluation criteria in solid tumors: Revised RECIST guideline (version 1.1)\u2014Japanese JCOG edition.\u201dhttps://www.parexel.com/files/5214/0422/3830/MI_Brain_Metastases_White_Paper_JUN_14.pdf) will be used. Target lesions and nontarget lesions defined on gadolinium-enhanced MRI with a 3-mm maximum slice thickness.For assessment of brain metastasis, \u201cBrain Metastases from Solid Tumors: Implementing Response Assessments\u201d and 15.15 months (95% CI: 12.45\u201326.55 months), respectively.http://nshi.jp/ejs/onesurvmst/). Allowing for dropouts, 65 subjects will be enrolled.As mentioned above, the threshold median PFS and expected median PFS are estimated to be 3.0 and 5.3 months, respectively. Under these conditions and assuming a 2-sided significance level of 5% and power of 75% from 1 sample nonparametric tests for a median survival time, 61 subjects are required . The trial is subject to the supervision and management of the Ethics Committee.32, the permissible starting dose of docetaxel for East Asian patients is 60\u200amg/m2. Therefore, we are using this dose in our trial.Although the REVEL study showed efficacy and tolerability of the treatment regimen with starting dose of ramucirumab 10\u200amg/kg and docetaxel 75\u200amg/mFor assessment of the tumor response, we are using 2 sets of guidelines: Response Evaluation Criteria in Solid Tumors (RECIST) for extracranial lesions, and \u201cBrain Metastases from Solid Tumors: Implementing Response Assessments\u201d for intracranial lesions. Patients who received only SRS or do not need treatment for local control of their brain tumors are included in this study; therefore, the brain lesions of most of the participants are <3\u200acm in diameter and amenable to SRS. Brain tumors initially >1\u200acm in diameter can be evaluated according to the RECIST guideline. On the contrary, \u201cBrain Metastases from Solid Tumors: Implementing Response Assessments\u201d is suitable for evaluating brain tumors >5\u200amm in diameter.4To the best of our knowledge, this study is the first clinical trial of ramucirumab combination therapy for NSCLC with brain metastasis. Our study may provide new evidence about brain metastasis treatment in patients with NSCLC.The authors thank the patients, their families, and all investigators involved in this recent study.Conceptualization: Keiko Tanimura, Junji Uchino.Formal analysis: Kenichi Yoshimura.Funding acquisition: Junji Uchino.Investigation: Keiko Tanimura, Nobuyo Tamiya, Yoshiko Kaneko, Tadaaki Yamada.Supervision: Koichi Takayama."} +{"text": "Twist1 is a master regulator of epithelial mesenchymal transition and carcinoma metastasis. Twist1 has also been associated with increased malignancy of human glioma. However, the impact of inhibiting Twist1 on tumorigenicity has not been characterized in glioma models in the context of different oncogenic transformation paradigms. Here we used an orthotopic mouse glioma model of transplanted transformed neural progenitor cells (NPCs) to demonstrate the effects of Twist1 loss of function on tumorigenicity. Decreased tumorigenicity was observed after shRNA mediated Twist knockdown in HPV E6/7 Ha-RasV12 transformed NPCs and Cre mediated Twist1 deletion in Twist1 fl/fl NPCs transformed by p53 knockdown and Ha-RasV12 expression. By contrast, Twist1 deletion had no effect on tumorigenicity of NPCs transformed by co-expression of Akt and Ha-RasV12. We demonstrated a dramatic off-target effect of Twist1 deletion with constitutive Cre expression, which was completely reversed when Twist1 deletion was achieved by transient administration of recombinant Cre protein. Together these findings demonstrate that the function of Twist1 in these models is highly dependent on specific oncogenic contexts of NPC transformation. Therefore, the driver mutational context in which Twist1 functions may need to be taken into account when evaluating mechanisms of action and developing therapeutic approaches to target Twist1 in human gliomas. Glioblastomas are lethal within 12\u201315 months after diagnosis in the majority of adult patients. This dismal prognosis is attributed in large part to highly invasive growth and capacity for resident glioma stem-like cells (GSCs) to drive tumor formation and progression, the latter a consequence of profound resistance to current standard of care therapies. Therefore the identification and inhibition of mechanisms which coordinately promote glioma cell invasiveness and stem cell phenotypes could be of great biologic and clinical relevance.in vitro [We previously reported that TWIST1 (TW), a critical regulator of epithelial mesenchymal transition (EMT), metastasis and stem cell phenotypes in carcinomas, also promotes mesenchymal change and invasion in glioma cells and enhances self-renewal of human glioma stem-like cells (GSC) in vitro . Althougin vitro . We and in vitro \u20136. Therein vivo mouse cancer models have shown that TW function is a critical downstream effector for malignant phenotypes generated by multiple oncogenic pathways [in vitro and in vivo by overriding oncogene induced senescence [Numerous pathways \u201314. Collnescence , 15 and nescence . In a monescence . While tTherefore, we employed our previously reported syngeneic mouse glioma model , 5 to in5 V38 cells into syngeneic hosts. At 40 days after implantation the mean tumor volume for V38 shTW derived tumors (0.2 cm3) was significantly reduced compared with those generated from V38 shScr cells but HPV per se is not pathogenic in human GBM. Therefore, we next employed additional transformation paradigms to test the effect of TW loss of function on tumorigenicity.Using previously generated and characterized HPV/Ras transformed NPCs derived from 3 month-old mouse forebrain we verif) Figure . The slo) Figure . These sIn the above experiments using shRNA we achieved partial knockdown of TW. In order to ascertain the absolute functional requirements of TW in transformed NPCs we employed a TW floxed transgenic model in which we could achieve complete knockout of TW expression. To provide a surrogate reporter of TW recombination and knockout we crossed TW floxed transgenic mice with mTmG mice (TWflox:mTmG). After exposure to cre recombinase and successful recombination at loxP sites, cells harboring the mTmG locus convert from Tomato red membrane fluorescence to GFP membrane fluorescence. This provides a readout for cre-mediated recombination at the TW loxP loci as well. Neural progenitor cells (NPCs) were isolated from the forebrain of 3 month old Twflox:mTmG transgenic mice Figure as previp = 0.0027). Furthermore AR cell-derived tumors exhibited hallmark pathologic features of human GBM including pseudopalisading necrosis, which was not observed in tumors derived from HPV/Ras transformed cells . Upon transformation we observed a marked upregulation of TW as shown above Figure . To delen Figure . The pren Figure and co-cn Figure . We concn Figure . To deten Figure . To testn Figure . Unexpecn Figure . This reTo address potential off-target effects of constitutive cre recombinase expression we analyzed the efficacy of transient exposure to recombinant cre protein in cell cultures to generate recombination. With 5 uM recombinant cre protein in culture we achieved 72% conversion rate from dTomato to GFP after 6 days in culture . With sein vitro by western blot after transformation with shP53/Ras compared normal cells with the pre-implanted shP53/Ras transformed NPCs. In 3 of 4 tumor derived cell isolates TW expression was increased at approximately 3, 6, and 6.5-fold compared with parental cells with one cell line exhibiting no change . Paraffin embedding, sectioning and hematoxylin and eosin staining were performed at the Histology Core of the University of Washington. Unstained slides were de-paraffinized and after blocking were incubated with Ki67 rabbit antibody and goat anti-rabbit secondary antibody conjugated with HRP. Antibody binding was visualized with diaminobenzidine (DAB) (Vectorlab) and counterstained with hematoxylin. DAB positive cells and total number of cells were counted in random fields of view. Labeling index is presented as mean percent of positive cells counted in 5 independent tumors in each group of animals.6 cells were plated on laminin-coated glass coverslips and placed in a 24-well dish in proliferation media. After 24 hours, cells were fixed with 4% paraformaldehyde at 37\u00b0C for 10 minutes, rinsed three times with PBS, and blocked for 1 hour in PBS with 0.1% Triton X-100 and 50% Problock. Fixed cells were then subject to overnight incubation at 4\u00b0C with primary antibodies 10% problock, 0.1% Triton-X in PBS including anti-Nestin mouse monoclonal , anti-SOX2 goat polyclonal , and anti-Olig2 rabbit polyclonal antibody and eluted with sterile water. Producing cells were transfected with Turbofect (Thermofisher) according to manufacturer's protocols in serum free DMEM/F12 media. For lentiviral production packaging DNA (Addgene) was co-transfected with vector harboring transgene of interest. After cell growth overnight in DMEM/F12 media supplemented with 10% FBS, cells were washed with PBS and serum free DMEM/F12 was added for 24\u201336 hours. Harvested viruses were filtered using syringe driven 0.45 \u03bcM PVDF filters. Target cells were transduced with virus diluted 1:1 with fresh media in the presence of 8mg/ml of Polybrene (Sigma). At least two sequential infection were performed followed by cell selection with appropriate antibiotics. After 10\u201312 days of selection cells were harvested and transgene expression was verified by Western blot.We previously transformed 3 month-old NPCs from C57Bl/6 females mouse forebrain using co-expression of HPVE6/7 and Ha-rasV12 or correTo enrich for specific TW wild-type or TW null genotypes cells were isolated on the basis of mTmG Cre reporter expression using Aria 3 at Cell Sorting Core of the University of Washington. To eliminate confounding effects from residual TW expressing cells we performed serial FACS (F1 and F2) to effectively eliminate TW+ cells after the F2 isolation.Western blots for TW and p53 were described , 5. EquaP53 stability assay was performed as described previously . Cells wRT-PCR and qRT-PCR were performed as previously described , 2. RNA Invasion assays were performed using matrigel-coated filters (BD Biosciences) as previously described except tin vivo studies. A Hamilton syringe and computer driven injector attached to the Stereotactic frame (Stoelting) were used for intracranial implantation of equal numbers (150 thousand cells/mice) of transformed NPCs into the indicated host strains to assess tumor formation and effect on survival . Injection coordinates relative to Bregma were 2 mm lateral, 1.5 mm anterior and 3 mm ventral. Procedures for anesthesia, analgesia, injection, post-op recovery and monitoring were performed according to approved IACUC protocol. Mice were sacrificed when moribund demonstrating following clinical manifestations: rough hair coat , hunched posture, decreased spontaneous activity, rapid shallow breathing or slow deep breathing, loss of body weight. For tissue collection, mice were deeply anesthetized with Avertin then transcardially perfused with PBS followed by 4% PFA. To assess the in vivo selection of TW+ and TW\u2013 cells with constitutive Cre expression, we injected nude hosts with 50:50 mixtures of TW+ and TW\u2013 cells as above and sacrificed animals at 10 and 20 days post injection without clinical manifestation.For the HPV/Ras studies where transformed NPCs were generated from C57Bl/6 mice we used 3mo syngeneic female C57Bl/6 mice as hosts. To mitigate logistic issues encountered in generating sufficient syngeneic hosts of the same age from the (129\u00d71/SvJ \u00d7 C57Bl/6) crosses we used immunocompromised nude mice (Taconic) for 4 cells. Cultures were analyzed by epi-fluorescent or confocal microscope at 2 and 6 days after seeding.Postnatal mouse brain slices were cultured according to established protocols and TW+ GFP expressing tumors were identified and dissected under a fluorescence dissecting microscope from unfixed tumor. GFP positive tumor fragments were digested with trypsin and plated in the media in the presence of a selection antibiotic to eliminate host-contaminating cells."} +{"text": "Purpureicephalus spurius) was assembled and characterized. The genome consists of 1,995 nucleotides and encodes two major proteins in opposing directions. This is the first evidence of BFDV infectivity and a complete genome sequence for this novel host.The complete genome sequence of beak and feather disease virus (BFDV) from a fledgling red-capped parrot ( Circoviridae family and a globally distributed pathogen for the Psittaciformes birds causing psittacine beak and feather disease (PBFD) .Beak and feather disease virus (BFDV) is a single-stranded DNA (ssDNA) virus from the e (PBFD) \u20134. The g (PBFD) \u2013, 5, 6. C (PBFD) \u2013. BFDV is (PBFD) \u2013, 9. In tBlood and feather samples were collected from a clinically suspected fledgling red-capped parrot . Routine HA and hemagglutination inhibition (HI) tests were performed as part Calyptorhynchus banksii) (GenBank accession no. KF385399) , while the cap gene showed 99% pairwise match with an isolate from a ringneck parrot (accession no. KF688549). The overall nucleotide identity of the new BFDV isolate ranges from 92 to 99% compared to the BFDV genomes available in GenBank comprises 1,995 nucleotides (nt), with a G+C content of 53.5%. Similar to other BFDV genomes, the basic structure includes two major open reading frames (ORFs), ORF1 nt 14 to 1000) and ORF2 (nt 1235 to 1984), containing genes encoding Rep and Cap, respectively. Preliminary BLASTn analysis to 1000 GenBank . This isKX449321.The complete genome sequence of BFDV has been deposited at GenBank under the accession no."} +{"text": "The purpose of this study was to assess the long-term prognostic value of cardiovascular magnetic resonance (CMR) stress perfusion in patients with atrial fibrillation who had suspected and known coronary artery disease (CAD) at initial stress CMR.130 consecutive patients with atrial fibrillation referred for perfusion stress CMR using either adenosine or regadenoson were followed for hard cardiovascular events defined as cardiac death or non-fatal myocardial infarction (MACE). Ischemia was defined as new onset of perfusion defects in at least two myocardial segments (positive test). Multivariable Cox regressions for MACE were performed to determine the prognostic value of CMR stress perfusion.Hard cardiac events occurred in 4 (3.1%) patients during the follow-up period (mean: 21 \u00b1 17 months). Patients without inducible perfusion defects (ischemia) experienced a substantially lower cumulative hard cardiovascular event rate (1%) than in patients with ischemia (9.1%) (p = 0.035) after 5 years (see Kaplan-Meier-curve).CMR stress perfusion in patients with atrial fibrillation can accurately identify patients, who are at increased risk for cardiac death and myocardial infarction, separating them from those with normal findings, who have a very low risk for future cardiac events."} +{"text": "Zfp148, ZBP-89, BFCOL, BERF1, ht\u03b2) interacts physically with the tumor suppressor p53, but the significance of this interaction is not known. We recently showed that knockout of Zfp148 in mice leads to ectopic activation of p53 in some tissues and cultured fibroblasts, suggesting that Zfp148 represses p53 activity. Here we hypothesize that targeting Zfp148 would unleash p53 activity and protect against cancer development, and test this idea in the Min/+APC mouse model of intestinal adenomas. Loss of one copy of Zfp148 markedly reduced tumor numbers and tumor-associated intestinal bleedings, and improved survival. Furthermore, after activation of \u03b2-catenin-the initiating event in colorectal cancer-Zfp148 deficiency activated p53 and induced apoptosis in intestinal explants of Min/+APC mice. The anti-tumor effect of targeting Zfp148 depended on p53, as Zfp148 deficiency did not affect tumor numbers in Min/+APC mice lacking one or both copies of Trp53. The results suggest that Zfp148 controls the fate of newly transformed intestinal tumor cells by repressing p53 and that targeting Zfp148 might be useful in the treatment of colorectal cancer.The transcription factor Zinc finger protein 148 ( Zfp148, ZBP-89, BFCOL, BERF1, ht\u03b2) interacts physically with the tumor suppressor p53, but the significance of this interaction is not known [Zfp148 deficiency leads to ectopic activation of p53 in mice and cultured fibroblasts, suggesting that Zfp148 represses p53 activity [Zfp148 leads to respiratory distress and partial neonatal lethality in mice that are caused by proliferative arrest of pulmonary cells, and to premature senescence in cultured mouse embryonic fibroblasts. The phenotypes are rescued by deletion of one or two copies of Trp53 (the gene encoding mouse p53). Moreover, loss of one copy of Zfp148 reduces proliferation of tissue macrophages and atherosclerosis in \u2212/\u2212Apoe mice by increasing p53 activity [Zfp148 are fertile and healthy [The transcription factor Zinc finger protein 148 gene [APC mutations give rise to adenomatous polyps that progress to carcinomas, a process driven by additional mutations including p53-mutations [Min/+APC mouse harbours a mis-sense mutation in the APC locus which leads to the production of a truncated APC protein [Min/+APC mice develop numerous adenomatous polyps, predominantly in the small intestine, which leads to intestinal bleedings, anemia and death at about 30 weeks of age [Several studies show that Zfp148 is required for the integrity of intestinal epithelium suggesting a possible link to colorectal cancer (CRC) -8. MoreoPC) gene . APC mututations . The APC protein . The muts of age . The modZfp148 deficiency activates p53 raises the possibility that Zfp148 promotes cancer by repressing p53 activity. In line with this, high expression of Zfp148 in oesophageal squamous cell cancer and clear cell renal cell carcinoma correlates with cancer progression and poor prognosis [Min/+APC model of CRC, and hypothesize that reduced expression of Zfp148 confers protection against intestinal adenomas by unleashing p53 activity. For this, we crossed Min/+Apc mice with mice lacking Zfp148 (gt/gtZfp148 and gt/+Zfp148) and studied the development of intestinal adenomas.Our finding that rognosis , 16. Howgt/+Zfp148 and gt/gtZfp148 mice on the Min/+APC genetic background to investigate if Zfp148 is involved in intestinal adenoma formation. Intestines from 12 week old mice were sectioned and stained with haematoxylin and eosin for evaluation of tumor development. Deletion of one or two copies of Zfp148 markedly reduced tumor numbers compared to Min/+APC controls, without significantly altering the anatomical distribution of tumors incorporation, expression of proliferating nuclear cell antigen (PCNA), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), there was no difference in cell proliferation or apoptosis of tumor cells or normal crypt epithelial cells between 12 weeks old gt/+Zfp148 mice and controls, respectively compared to age-matched controls, raising the possibility that Zfp148 deficiency reduces progression of advanced tumors of known signaling pathways for 6 and 20 hours. Phosphorylation of \u03b2-catenin by GSK3\u03b2 is required for proteasomal destruction of \u03b2-catenin and inhibition of GSK3\u03b2 activates \u03b2-catenin.Tumor initiation is triggered by loss of the second gt/+Zfp148 mice at 20 hours after GSK3\u03b2 inhibition in human CRC was recently investigated . In famiZfp148 reduces tumor initiation is therefore clinically interesting. Since mice lacking one copy of Zfp148 appear to be healthy and have a normal lifespan, our finding suggests that therapeutic targeting of Zfp148 could reduce the incidence of CRC by increasing p53 activity without causing detrimental side effects. However, transcription factors such as Zfp148 are not ideal drug targets although significant progress has been made in this field. A more realistic approach may be to target other proteins in the Zfp148-p53 pathway.Cancer chemoprevention is often cited as an effective strategy to reduce cancer mortality. Our finding that global deletion of one copy of Wip1 suppresses MinAPC-driven polyposis by increasing p53 activity in response to constitutively activated \u03b2-catenin [The mechanism by which Zfp148 regulates p53 activity is not known , 3. The -catenin , it is pZfp148 deficiency reduces tumor formation in the Min/+APC model. CRC is one of the most prevalent cancer forms in the Western world and a leading cause of cancer-related death. Mutations in the APC gene is a major causative event of CRC in humans [Min/+APC model is of clinical interest. Because deletion of Zfp148 reduces the initiation of tumors, preventive targeting of Zfp148 may be efficient to reduce the incidence of CRC and thereby CRC mortality.In this study we show that n humans and idenMin/+Apc and tm1TyjTrp53 (Trp53-/) mice were obtained from The Jackson Laboratory and gt/+Zfp148 mice were produced by us [ed by us . The micMin/+/Zfp148+/+APC, Min/+/Zfp148gt/+APC and Min/+/Zfp148gt/gtAPC mice. Mice were dissected, the intestines were removed and separated into four segments; colon and three segments of the small intestines. The segments were rinsed in PBS and prepared using the Swiss roll technique [Adenoma frequency was compared in 12 week echnique . TissuesMice were sacrificed when they became moribund, defined as when they became listless, their haematocrit was below 20%, or their bodyweight was reduced by 15%.Blood was drawn from the tail into EDTA-coated tubes . Samples were analysed on a Hemato analyser KX-21N (Sysmex) to determine haemoglobin, hematocrit and leukocyte count.Faecal blood was detected by using Hemoccult test for faecal blood (Beckman-Coulter).Intestinal tissues were fixed in 4% formaldehyde, imbedded in paraffin and sectioned in 5 um thin sections. Staining was performed using standard protocol. Slides were boiled for 10 min in Citric acid to unmask epitopes. Primary antibody anti-PCNA was diluted 1:1000 and secondary antibody was diluted 1:500 and primary antibody anti-\u03b2-catenin , was diluted 1:200 and visualized with Vectastain Elite ABCkit, Peroxidase, Rabbit IgG, (Vector Laboratories) as before .\u00ae Fluorescence In Situ Apoptosis Detection Kit (Millipore). Cell proliferation was evaluated with 5-Bromo-2\u2032-deoxy-uridine (BrdU) Labeling and Detection Kit I (Roche) 2h after intraperitoneal injection of BrdU (75mg/kg) according to manufacturer's description and as before [Apoptosis was evaluated with Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) using the ApopTags before .\u00ae WT PLUS Reagent Kit User Manual . GeneChip\u00ae ST Arrays (GeneChip\u00ae XXX Gene 2.0 ST Array) were hybridized for 16 hours in a 45\u00b0C incubator, rotated at 60 rpm. According to the GeneChip\u00ae Expression Wash, Stain and Scan Manual the arrays were then washed and stained using the Fluidics Station 450 and finally scanned using the GeneChip\u00ae Scanner 3000 7G. The gene expression data set has been deposited in the GEO repository with accession number GSE77773.Total RNA was extracted from crypt-enriched tissues that were isolated by laser microdissection (PALM) from 10\u03bcm thick cryosections of OCT-embedded and snap frozen Swiss roll preparations of small intestines . RNA concentration was measured with ND-1000 spectrophotometer and RNA quality was evaluated using the Agilent 2100 Bioanalyzer system . 250 nanograms of total RNA from each sample were used to generate amplified and biotinylated sense-strand cDNA from the entire expressed genome according to the GeneChiphttp://www.affymetrix.com) using the robust (RMA) method first suggested by Li and Wong in 2001 [http://www.r-project.org) using packages available from the Bioconductor project (www.bioconductor.org). In order to search for the differentially expressed genes between Min/+APC and the gt/+Zfp148 groups an empirical Bayes moderated t-test was then applied [The raw data was normalized in the free software Expression Console provided by Affymetrix using gene sets from the KEGG (Kyoto Encyclopedia of Genes and Genomes) repository (c2.cp.kegg.v4.0.symbols.gmt) and default settings.2. The mixture containing the tissue was transferred to Eppendorf tubes and centrifuged at 14,000 rpm for 5 min. The tissue was collected after removal of the supernatant and used for analysis of mRNA or protein expression. All explant experiments were performed on Min/+APC background.One 2 cm segment of the distal ileum was collected, as previously described . IntestiTaqMan assays were performed as described using TaProtein levels was determined as previously described with antgt/++/+Trp53) or laser microdissected (gt/+Trp53+/Zfp148) tumors and subjected to Sanger sequencing. Each amplicon was sequenced twice and scanned for deviations from the refseq sequence. Sequencing was performed by Beckman Coulter Genomics using primers listed in Exons 5-8 of mouse Trp53 were PCR amplified from genomic DNA isolated from dissected . Investigators were blinded to the genotype and values are presented as mean \u00b1 SEM. A p-value of < 0.05 was considered statistically significant.Statistics were performed with non-parametric Mann-Whitney test for tumor count; Chi Square test for tumor distribution and histopathological grade; ANOVA test for haemoglobin;"} +{"text": "Fetuses and infants with congenital heart disease (CHD) have delayed brain maturation and lower brain volumes (BV) compared to normal [1-4]. To understand the impact of CHD and cardiac surgery on brain maturation, we performed serial brain MRI studies in patients with common cyanotic CHD before and after birth.Post-natal brain MRI were performed without sedation in 24 infants with common CHD before and after the cardiac surgery on a Siemens Avanto 1.5T system (Erlangen) after hospital IRB approval. 18 of 24 subjects also had fetal MRI using previously described technique . The norThe cohort (n = 24) consisted of patients with transposition of the great arteries (TGA) with intact ventricular septum ; TGA with ventricular septal defect , hypoplastic left heart syndrome ; tricuspid atresia , pulmonary atresia . The TGA/IVS group had normal brain growth after birth and surgery Figure . Howeverin utero until surgery. Delayed repair leaves them exposed to adverse brain hemodynamics for a longer time. The reversal of normal decline in T2 and ADC in TGA indicates additional pathological process in these brains predisposing them to WM injury during cardiac surgery.Infants with TGA/VSD have the most immature brains among common cyanotic CHD probably related to low CDO2"} +{"text": "This was a prospective study designed to evaluate the impact of thyroid function abnormalities on reproductive hormones during menstrual cycle in HIV infected females at Nnamdi Azikiwe University Teaching Hospital Nnewi, South-East Nigeria.The study randomly recruited 35 Symptomatic HIV infected females and 35 Symptomatic HIV infected females on antiretroviral therapy (HAART) for not less than six weeks from an HIV clinic and 40 apparently heathy control females among the hospital staff of NAUTH Nnewi. They were all premenopausal females with regular menstrual cycle and aged between 15\u201345 years. Blood samples were collected at follicular and luteal phases of their menstrual cycle for assay of Thyroid indices and Reproductive indices using ELISA method.The result showed significantly higher FSH and LH but significantly lower progesterone (prog) and estrogen (E2) in the test females compared to control females at both phases of menstrual cycle (P<0.05). There was significantly lower FT3 but significantly higher TSH value in Symptomatic HIV females (P<0.05). FSH, LH and TSH values were significantly lowered while prog and FT3 were significantly higher in Symptomatic HIV on ART compared to Symptomatic HIV females (P<0.05). FT3, FT4, Prog and E2 were inversely correlated while FSH and LH were positively correlated with duration of HIV infection in HIV females (P<0.05 respectively). There was a direct correlation between CD4+ count and FT3 while inverse correlation was found between CD4+ count and TSH levels (P<0.05).The present study demonstrated hypothyroidism with a significant degree of primary hypogonadism in Symptomatic HIV infected females at both follicular and luteal phases of menstrual cycle which tends to normalize on treatments. Contrastingly, nadir CD4+ T-cells (cells/\u03bcL) was not significantly different between symptomatic HIV infected females (212 \u00b1 109) and their corresponding females on HAART (296 \u00b1 98) (P>0.05). Menstrual irregularities percentage was higher in symptomatic HIV infected females 18(40%) as compared to symptomatic HIV infected females on HAART 28(25.7%) while 34(34.3%) of HIV females have regular menstrual cycles (P<0.05). There was no significant difference observed between symptomatic HIV females who were receiving zidovudine (NRTI), lamivudine (NRTI) and nevirapine (NNRTI) and those who were receiving Stavudine (NRTI), Lamivudine (NRTI), and Nevirapine (NNRTI) (P>0.05) See .FT3, FT4, Prog and E2 were inversely correlated while FSH and LH were positively correlated with duration of HIV infection in HIV females (P<0.05 respectively) . There wThe mean (\u00b1SD) serum FSH, LH and Prolactin Levels in Symptomatic HIV females were not significantly different between follicular and luteal phases of menstrual cycle (P>0.05 respectively). Similarly, there was no significant difference in the mean LH and Prolactin concentrations between follicular and luteal phases of menstrual cycle in Symptomatic HIV female subjects on HAART (P>0.05) but the mean FSH value dropped significantly at luteal (13.5\u00b12.5) phase compared with follicular (18.2\u00b110.4) phase of menstrual cycle in Symptomatic HIV females on HAART (P<0.05). The mean serum FSH and LH values in Control females were significantly higher at follicular phase than at luteal phase of menstrual cycle (P<0.05 respectively). There was no significant difference in the mean prolactin value between follicular (18.3\u00b14.4) and luteal (18.1\u00b13.4) phases of menstrual cycle in Control female subjects (P>0.05).The mean FSH and LH were significantly higher in Symptomatic HIV compared with the corresponding value in the Control female subjects (7.9\u00b16.3) (P<0.05). FSH was significantly higher at follicular phase in Symptomatic HIV on HAART (18.2\u00b110.4) compared with corresponding value in the Control female subjects (7.9\u00b16.3) (P<0.05). The mean FSH and LH concentration at luteal phase of menstrual cycle were significantly higher in Symptomatic HIV and Symptomatic HIV on HAART compared with corresponding value in the Control female subjects (4.4\u00b11.4) (P<0.05 in each case).The post hoc analysis showed significant drop in the mean LH value at follicular phase of menstrual cycle in Symptomatic HIV females on HAART (10.6\u00b15.4) compared with the corresponding value in Symptomatic HIV female subjects (18.0\u00b116.6) (P<0.05) See .The mean (\u00b1SD) serum progesterone and estradiol concentrations in Symptomatic HIV females were not significantly different between follicular and luteal phases of menstrual cycle (P>0.05 respectively). In Symptomatic HIV females on HAART, there were no significant difference in the mean progesterone and estradiol concentrations between the follicular and luteal phases of menstrual cycle (P>0.05). But the mean serum progesterone and estradiol concentrations dropped significantly at follicular phase compared with the luteal phase of menstrual cycle in Control female subjects (P<0.05 respectively).When the mean progesterone concentrations at follicular and luteal phases of menstrual cycle were compared between the Control group and the Test groups, the mean progesterone concentration dropped significantly in Symptomatic HIV and Symptomatic HIV on HAART compared with the corresponding values in the Control female subjects (P<0.05 respectively). Similarly, When the mean progesterone concentrations at luteal phase of menstrual cycle were compared between the Control group and the Test groups, the mean progesterone and estradiol values dropped significantly in Symptomatic HIV and Symptomatic HIV on HAART compared with corresponding values in the Control female subjects (P<0.05 respectively) See .The mean (\u00b1SD) serum FT3, FT4 and TSH levels in Symptomatic HIV females were not significantly different between follicular and luteal phases of menstrual cycle (P>0.05 respectively). There were no significant difference in the mean serum FT3 and FT4 concentrations (ng/ml) between follicular and luteal phases of the menstrual cycle in Symptomatic HIV females on HAART (P>0.05). On the contrary, the mean serum TSH value was significantly higher at follicular phase (3.26\u00b11.88) compared with luteal phase (1.24\u00b10.39) of menstrual cycle in Symptomatic HIV females on HAART (P<0.05). In Control female subjects, there were no significant difference in the mean serum FT3, FT4 and TSH concentrations (ng/ml) between follicular and luteal phases of menstrual cycle (P>0.05).The mean FT3 at follicular and luteal phases dropped significantly in Symptomatic HIV females compared with the follicular and luteal values in Control female subjects (P<0.05 respectively). The post hoc analysis showed significant drop in the mean FT3 value (ng/ml) at follicular phase of menstrual cycle in Symptomatic HIV females compared with follicular value in the Symptomatic HIV females on HAART while the mean TSH was significantly higher in Symptomatic HIV females and Symptomatic HIV females on HAART at follicular and luteal phases compared with the corresponding values in the Control female subjects (P<0.05 respectively).The post hoc analysis dropped significantly in the mean TSH value at luteal phase of menstrual cycle in Symptomatic HIV females on HAART (1.24\u00b10.39) compared with Symptomatic HIV females (4.56\u00b11.17) (P<0.05) See .The present study showed that the mean serum levels of FSH, LH, progesterone and estradiol in Symptomatic HIV were not significantly different between follicular and luteal phases of menstrual cycle. This contrasts the observation in apparently healthy subjects where differences in hormonal levels exist between the two phases of the menstrual cycle. However, the significantly higher estradiol at luteal phase compared to follicular phase of menstrual cycle suggests some degree of impact of treatment on these subjects. FSH and LH are usually higher at the follicular phase and peak at the mid cycle to enable ovulation to take place while PrIn normal healthy women, Ovarian follicles, from the pool of resting primordial follicles either continue to grow from preantral to antral follicles due to survival signals, such as gonadotrophins and growth factors, or degenerate and die by the process of follicular atresia. Expressions of hormones and growth factors have been shown to regulate the destiny of the ovarian follicle. In humans, disorders of the thyroid gland are responsible for a dysregulation of the hypothalamus, pituitary, gonadal axis, and hypothyroidism is associated with oligomenorrhea . The folet al. reported that hypothyroidism does not influence the classical preovulatory patterns of LH and FSH secretion in rats [However, in the present study, the serum levels of FSH and LH were significantly higher while progesterone and estradiol levels were significantly lower in Symptomatic HIV, Symptomatic HIV on HAART at both follicular and luteal phases of menstrual cycle. The implication of grossly increased FSH with reduced progesterone is primary ovarian failure. This means that the anterior pituitary is overworking itself to stimulate a poorly responding ovary. The abnormally low progesterone signifies hypogonadism and may lead to menstrual and reproductive failure . Normall in rats .The thyroid hormone plays a vital role in all physiological activities in humans including menstrual functions in females. Increased thyroid function (hyperthyroidism) may lead to premature menstruation or precotious puberty, menorrhagia or hypermenorrhoea whereas reduced thyroid function (hypothyroidism) may lead to delayed menstruation or oligomenorrhoea and pregnancy loss , 32. ThiFurther analysis showed that seventeen out of twenty (85%) Symptomatic HIV female subjects had significantly higher FSH and LH levels while progesterone and estradiol were being secreted below the lower limit of normal. This strongly indicates a state of reduced reproductive hormones causing hypogonadism. The significantly reduced levels of progesterone and estradiol probably as a result of HIV infections send a pThe reduction of FSH, appreciation in the level of progesterone with the insignificant difference in the levels of FT3 and TSH observed in HIV subjects on HAART suggests stimulatory effects of the treatment on the gonads and possible reduction on the incidence of hypothyroidism with intact negative feedback mechanism thereby resulting in the restoration of the gonadal functions showing some beneficial effects and a tendency to return to normal. This may reduce the incidence of menstrual abnormality and infertility. Studies have demonstrated a positive correlation between TSH and PRL in hypothyroid women . In a prHIV disease is associated with opportunistic infections which are linked with increased levels of pro- inflammatory cytokines. The later may be responsible for the hypogonadism observed in HIV, which calls for further investigations. However, prolonged use of HAART (protease inhibitors) has been associated with thyroid abnormalities such as Hashimoto\u2019s thyroiditis which results to a kind of immune reconstitution \u201340. Howeet al found that only stavudine treatment and low CD4 count were statistically associated with hypothyroidism [et al. found that TSH levels were negatively correlated with CD4+ count nadir [et al., [et al., [et al. suggested that immune reconstitution was more likely to protect the thyroid function than impair it [The present study did not find any significant association between age and thyroid hormones nor reproductive hormones assayed in the affected women. Thyroid hormones were not significantly associated with viral load, nadir CD4+ count and duration of HAART. Previous report has shown that FT3 and FT4 were related to the state of HIV infection and are potential biomarkers of HIV progression . Beltralyroidism while Mant nadir which wa[et al., and Shuj[et al., ). It is [et al., , 43, 44 [et al., while Simpair it .et al. in their similar study reported hypogonadism in 6%, 40%, and 50% of asymptomatic HIV positive, symptomatic HIV positive and AIDS patients respectively [et al. reported that primary hypogonadism was more common cause of hypogonadism [The present study reported that the percentage of menstrual irregularities was higher in symptomatic HIV infected females 18(40%) as compared to symptomatic HIV infected females on HAART 28(25.7%). This was supported by the significant hypogonadism reported in the present study. Dobs ectively . Meena egonadism , 49 whilgonadism . Primarygonadism , 53.In conclusion, the present study demonstrated hypothyroidism with significant degree of primary hypogonadism in Symptomatic HIV infected females at both follicular and luteal phases of menstrual cycle which tends to normalize on treatment. A counselling module for reproduction and routine screening for thyroid function is therefore advocated to be included in voluntary counselling of HIV subjects to reduce the incidence of reproductive and thyroid dysfunctions in affected women.S1 Questionnaire(DOC)Click here for additional data file."} +{"text": "Anopheles gambiae females, which are the dominant malaria mosquito vector in Sub Saharan Africa, it substantially shortens lifespan, prevents insemination and egg production, and significantly blocks Plasmodium falciparum development, three components that are crucial to malaria transmission. Modeling the impact of these effects on Anopheles population dynamics and Plasmodium transmission predicts that disrupting steroid hormone signaling using 20E agonists would affect malaria transmission to a similar extent as insecticides. Manipulating 20E pathways therefore provides a powerful new approach to tackle malaria transmission by the mosquito vector, particularly in areas affected by the spread of insecticide resistance.The control of mosquito populations with insecticide treated bed nets and indoor residual sprays remains the cornerstone of malaria reduction and elimination programs. In light of widespread insecticide resistance in mosquitoes, however, alternative strategies for reducing transmission by the mosquito vector are urgently needed, including the identification of safe compounds that affect vectorial capacity via mechanisms that differ from fast-acting insecticides. Here, we show that compounds targeting steroid hormone signaling disrupt multiple biological processes that are key to the ability of mosquitoes to transmit malaria. When an agonist of the steroid hormone 20-hydroxyecdysone (20E) is applied to Plasmodium parasites. Here we show that when steroid hormone signaling is interrupted in female Anopheles mosquitoes, various aspects of their lifecycle are disrupted\u2014females produce and lay fewer eggs, do not mate successfully and die more rapidly. Furthermore, they become less likely to be infected by malaria parasites. When we model the impact of steroid hormone agonists on malaria transmission, we predict that these compounds would provide an important new tool against malaria, particularly in regions of widespread insecticide resistance.Mosquito control is the only intervention that can reduce malaria transmission from very high levels to close to zero. However, current mosquito control methods are severely threatened by the rapid spread of insecticide resistance in anopheline mosquito populations that transmit the malaria-causing Anopheles species that transmit human malaria remains the cornerstone of prevention and transmission reduction efforts , 61m(t),a, in the Ross-MacDonald model of malaria transmission [We consider interventions in settings with high 85%), moderate (45%), and low (5%) malaria prevalence pre-intervention, mediated by the number of bites per human per mosquito per day . Our quasmission , 8 as it5%, moderPlasmodium infection was estimated separately for the DBH-exposed groups and control groups using experimental data Click here for additional data file.S2 TableDaily mortality rates as calculated from the experimental results from (PDF)Click here for additional data file.S1 Fig(A) 10% ethanol/PBS (injection vehicle control), and (B) 38 mM 20E in 10% ethanol/PBS. (C) Topical application of 2 \u03bcg of DBH in 5% DMSO/acetone (0.4% w/v). The primary (1\u00b0F), secondary (2\u00b0F), and\u2014where visible\u2014tertiary (3\u00b0F) follicles, nurse cells (NC) and the oocyte (OOC) are indicated. Fragmented chromatin (indicating apoptosis) was labelled with FITC (green) using TUNEL. DNA was stained with DAPI (blue). Both 20E injection and DBH exposure induce extensive apoptosis in primary follicles (*) at 24h post treatment. The scale bar (bottom right) represents 50 \u03bcm.Intrathoracic injection of 138 nl of (TIF)Click here for additional data file.S2 FigAfter treatment with 3 DBH doses , females who developed oocysts following an infectious blood meal showed similar intensity of infection compared to controls. Number of individuals (n) is listed below each DBH condition.(TIF)Click here for additional data file.S3 Fig(A) The fraction of mosquitoes exposed increases non-linearly with the intervention applied: no effect (gray), DBH alone (pink lines), insecticide alone (blue lines). (B) The daily adult survival curve and (C) cumulative survival curve as estimated from the experimental data for the model (modeled to reflect (D) The daily mortality in the larval stage is a sigmoidal function (solid line) of the larval population size. Previous models [E) The daily risk of infection by mosquitoes biting infected humans increases quickly at low prevalence in humans and reaches a maximum of 10%.s models , 66 have(TIF)Click here for additional data file.S4 Fig(A) The adult mosquito population varies non-linearly under increasing coverage with different DBH doses (pink lines) or insecticide efficacy (blue lines). The vertical yellow bar indicates 85% coverage for which the age distribution of the population is considered in (B). The population size shown is relative to the total female population in the absence of any interventions. (B) The age distribution of female mosquitoes in the presence of 2.0 \u03bcg DBH (pink) or the absence of any intervention (gray) at 85% coverage, indicated by a yellow bar in (A) and (C). The highlighted days in the x-axis indicate the age range of mosquitoes that are old enough to transmit malaria if infected. (C) The potentially infectious adult mosquito population under increasing levels of coverage with different DBH doses (pink lines) or insecticide efficacy (blue lines). This includes females at least 12 days after a blood meal, and in the case of DBH exposure, a proportion of these females are excluded due to reduced Plasmodium susceptibility. The yellow bar indicates 85% coverage, for which the age distributions of the population are considered in (B). Without intervention, the proportion of mosquitoes 14 days or older (black line) is 0.22. Insecticide of 60%, 80%, or 100% efficacy and DBH of experimentally determined efficacy (efficacy are used(TIF)Click here for additional data file.S5 Fig(A) Increases in egg batch size are associated with increases in egg and larval populations while adult populations show non-linear increases with a peak population size when egg batch size is small. However, larger egg batches are observed in field and experimental settings, likely due to the steep decline in adult population just below the optimal batch size. In particular, due to the dependence on climatic factors, the egg batch size must be substantially larger to avoid population crashes through stochastic drops in egg batch size. (B) The older adult mosquito population, i.e. at least 12 days after their first feed, under increasing levels of coverage with different DBH doses (pink lines) or insecticide efficacy (blue lines). The yellow bar indicates 85% coverage, for which the demographics of the population are considered in (TIF)Click here for additional data file.S6 FigM) become exposed (EM) by feeding on an infectious human. The force of infection from humans to mosquitoes is a function of the proportion of humans who are infectious (M). We use a Susceptible-Infectious model to follow malaria in humans. Susceptible humans (SH) become infectious (IH) immediately after an infectious mosquito bite. The force of infection from mosquitoes to humans is a function of: (i) the proportion of mosquitoes which are infectious, (ii) the number of bites per mosquito per human per day, and (iii) the per-bite probability of transmission. The mean recovery period for infectious humans is 75 days.We use a Susceptible-Exposed-Infectious model to track malaria in mosquitoes. Susceptible mosquitoes (Sfectious . The lat(TIF)Click here for additional data file.S7 Fig(A) low, (B) moderate, or (C) high transmission setting. Effectiveness at 2 \u03bcg DBH (solid pink line) was greater than or comparable to 100% insecticide efficacy, while 0.5 \u03bcg (dashed pink line) and 0.125 \u03bcg DBH (dotted pink line) had effectiveness similar to that of 80% (dashed blue line) or 60% (dotted blue line) insecticide efficacy, respectively. The maximal DBH efficacy considered is our experimentally determined effects on egg development, mating, mortality, and Plasmodium susceptibility and efficacy (line style) as determined from dose response experiments for tibility .(TIF)Click here for additional data file."} +{"text": "Objective. Retrospective studies have found that noninvasive encapsulated follicular variant of papillary thyroid cancer (EFVPTC) exhibits highly indolent clinical behavior. We studied the clinical features of our patients with noninvasive EFVPTC tumors culled from a community endocrine surgical practice registry over the past four years. Methods. We interrogated the Memorial Center for Integrative Endocrine Surgery (MCIES) Registry for all recorded encapsulated follicular variant of papillary cancer pathologic diagnoses. We identified a subgroup of patients without capsular or vascular invasion and studied their clinical characteristics. Results. Thirty-seven patients met inclusion and exclusion criteria. The typical patient was young and female. Nodules averaged 3.1\u2009cm in greatest dimension by ultrasound evaluation. Thirteen patients were found to have synchronous malignancies elsewhere in the thyroid (35%). At the time of this writing, we have not seen a clinical recurrence in any of our 37 noninvasive EFVPTC patients. Conclusions. Early clinical follow-up data suggests that the majority of noninvasive EFVPTC tumors exhibit indolent behavior, but clinical decision-making with regard to completion thyroidectomy, central lymph node dissection, and adjunctive radioiodine therapy often depends on the amount and type of synchronous thyroid cancer detected elsewhere in the thyroid gland and the central neck. The incidence of thyroid cancer diagnoses has tripled over the past 30 years . EncapsuIn light of recent important changes in thyroid pathological classification, we sought to retrospectively evaluate our community endocrine surgical experience with noninvasive EFVPTCs over the past 45 months. Based on our clinical, ultrasound, and fine needle aspiration biopsy cytology data, we propose an algorithm for surgical management in suspected noninvasive EFVPTC patients.We queried the Memorial Center for Endocrine Surgery Registry for fully encapsulated follicular variant of papillary cancer diagnoses from January 2012 through September 2016. All these tumors were removed by a single surgeon, DNB, and had pathologic diagnoses rendered by a single pathology group serving our six-hospital consortium with a patient referral base encompassing South Florida and the Caribbean . Neck ultrasound evaluation was performed by AG or RMH for 36 out of 37 patients (97% of patients). Thyroid fine needle aspiration (FNA) biopsies were performed by AG or RMH in 15 thyroid nodules (40.5%) with 15 nodule FNAs (40.5%) performed by outside physicians. Seven nodules (19%) were not biopsied prior to removal. Cytology was read by Thyroid Cytopathology Partners of Austin, Texas, in 51% of cases and a combination of CBL and other community reference laboratory cytopathologists in 49%.Tumors in which the pathologist identified any vascular invasion or any capsular discontinuity or invasion were excluded from consideration in this report. Isolated subcentimeter encapsulated follicular variant of papillary tumors (EFVPTC) that were incidentally noted by the pathologist following surgery for another indication were excluded. In addition, tumors without ultrasound data were excluded (one patient).We systematically reviewed electronic medical records on each of the subjects to confirm that ultrasound, cytology, and pathology reports were concordant on a nodule-by-nodule basis.Twelve hundred and ten unique patients underwent thyroid surgery at the Memorial Center for Integrative Endocrine Surgery from January 2013 through September of 2016, of which 796 demonstrated nonincidental thyroid malignancies. Of these, 413 (52%) were follicular variant of papillary cancers (FVPTC). Eighty-eight (21%) of the FVPTC tumors were fully encapsulated (EFVPTC) and 50 (12%) were fully encapsulated with no evidence of capsular or vascular invasion and were considered noninvasive EFVPTC. In these fifty patients, neither capsular invasion, vascular invasion, nor papillary architectural features were identified in the primary tumor pathology report. Twelve of these tumors were excluded because they were subcentimeter incidental lesions discovered by our pathologists in thyroid glands removed for other reasons (symptomatic goiter or larger non-EFVPTC tumors). One tumor was excluded due to the absence of ultrasound data. Thus 37 out of 796 (4.6%) of our resected thyroid malignancies were found to meet our inclusion criteria as noninvasive encapsulated follicular variant of papillary thyroid cancers removed by our surgeon from 1/2013 to 9/2016. Thirty-seven tumors of interest were found in 37 patients.In our practice, the typical noninvasive EFVPTC tumor patient had an average age of 44 years. Twenty-nine of the 37 patients were female (78%).On neck ultrasound, noninvasive EFVPTC nodules had a mean diameter of 3.1\u2009cm in greatest dimension (range 0.8\u2009cm\u20136.9\u2009cm with a median size of 3\u2009cm). Nodules were evenly distributed throughout the thyroid with 18 (49%) in the right lobe, 17 (46%) in the left lobe, and 2 (5%) in the isthmus. Echogenicity was variable, with 9 (24%) read as hypoechoic, 21 (57%) read as isoechoic, and 7 (19%) described as heteroechoic on ultrasound. Thirty-three nodules (89%) were solid and 4 (11%) were described as partially cystic. Borders were sharp and well defined in 34 nodules (92%). Three nodules had hypoechoic halos (8%). Thirty-six nodules (97%) lacked intranodular calcifications, whereas intranodular arcuate calcification was described in one nodule. Twelve nodules (32%) demonstrated grade 1 vascularity as assessed by Power Doppler, 11 nodules had grade 2 vascularity (30%), and 14 nodules had grade 3 vascularity (38%). No nodules had grade 4 vascularity. The mean vascularity grade was 2.1.Thirty patients out of 37 (81%) underwent fine needle aspiration biopsy prior to surgery. Seven patients were referred directly to surgery based on sonographic findings alone. Eight nodules (27%) yielded benign cytological results (Bethesda 2), 14 (47%) nodules demonstrated atypia of uncertain significance (Bethesda 3), and 8 (27%) nodules were read as follicular neoplasm/suspicious for follicular neoplasm (Bethesda 4). Nondiagnostic, suspicious for malignancy, or frankly malignant cytological results were not encountered .The Afirma GEC\u00ae gene expression classifier was requested in eleven nodules with Bethesda 3 and three nodules with Bethesda 4 cytology. The GEC result was \u201csuspicious\u201d in all eleven Bethesda 3 nodules and in two of the three Bethesda 4 nodules. The Afirma GEC \u201cbenign\u201d Bethesda 4 nodule was removed due to its size (6.9\u2009cm) and suspicious sonographic features and 17 patients underwent lobectomy. Four patients who had an initial lobectomy underwent a completion thyroidectomy and 2 of these 4 patients had microscopic follicular variant of papillary cancer in the lobe opposite to the previously removed noninvasive EFVPTC tumor. Permanent postsurgical hypoparathyroidism and vocal cord dysfunction were not encountered in any of the 37 patients.On histologic examination, 13 out of 37 (35%) thyroidectomy specimens were found to contain additional incidental malignancies. These included 10 patients with incidental subcentimeter papillary thyroid cancers (PTCs) or follicular variant of papillary cancers (FVPTCs) contained one or more malignant lymph nodes with subcentimeter, noninvasive tumor deposits. In the first patient culled from a community-based endocrine surgery registry from January 2013 through September 2016.Our noninvasive EFVPTC patients were relatively young mean age 44) and predominately female with nodules that, on ultrasound, were large (3.1\u2009cm on average), hypo- to isoechoic, and mildly hypervascular with crisp boundaries in all 37 cases. Highly suspicious sonographic features such as microscopic calcifications or infiltrative borders were not seen in any of these lesions. Noninvasive EFVPTC nodules typically yielded indeterminate cytology results on fine needle aspiration biopsy (73% Bethesda Classification 3 or 4) with 12 of the 13 (92%) who underwent Afirma GEC testing classified as \u201csuspicious.\u201d Although we did not routinely perform mutational testing at our center from 1/2013 to 9/2016, two of our patients were found at outside institutions to have DNA alterations including a PAX8/PPAR gamma translocation and a low level NRAS mutation affecting 8% of thyrocytes, corresponding to an allelic frequency of 4%. Although we did not recut surgical specimens, our noninvasive EFVPTC patients' clinical, ultrasound, cytology, and molecular characteristics appear to resemble those of the NIFTP tumor patients described by Nikiforov et al. [4 and preInterestingly, in our patient group, molecular markers influenced surgical decision-making in 16 of the 17 patients in whom molecular testing was performed, while the other twenty patients had surgical decision-making driven by other factors such as family history, clinical symptoms, and, most importantly, ultrasound findings. Cytology evaluation was less helpful as all our patients who underwent fine needle aspiration biopsy (FNA) fell into Bethesda categories 2 (Benign), 3 (AUS/FLUS), or 4 (FN/SFN).Twenty out of 37 of our noninvasive EFVPTC patients (54%) underwent total thyroidectomy due to primary tumor size, tumor appearance, or ultrasound demonstrable contralateral disease. One of these twenty patients underwent a total thyroidectomy and a central lymph node dissection due to suspected central lymph involvement and subsequently received postoperative radioiodine therapy for remnant ablation when 4 central lymph nodes were found to contain microscopic cancer on pathologic examination with the largest nodal deposit measuring 0.4\u2009cm had synchronous thyroid cancers elsewhere in the thyroid gland. This is a higher than expected rate of thyroidectomy-detected occult microscopic carcinoma with the typical reported rate approximating 10% , many reIn the current endocrine surgical practice environment we lack prospective studies that provide guidance for the surgical management of patients with suspected noninvasive EFVPTCs and/or NIFTP tumors (collectively referred to as \u201cencapsulated follicular tumors\u201d). Our current approach is to assess all patients referred for thyroid nodule evaluation for characteristic ultrasound and fine needle aspiration features of encapsulated follicular tumors which include (1) nonhyperechoic lesion on ultrasound, (2) smooth, uniform, and uninterrupted margins, (3) no microscopic calcifications, (4) Bethesda 3 or 4 cytology, and (5) either a suspicious Afirma GEC or a mutation panel demonstrating a follicular tumor mutation, rearrangement, or fusion . In suspIn conclusion, because of a high risk of tumor multifocality, careful preoperative neck ultrasound and appropriate use of fine needle aspiration biopsy are essential in the management of suspected noninvasive EFVPTC tumors. The extent of surgery should be guided by ultrasound findings, biopsy results and clinical judgement. Molecular testing with Afirma GEC is often \u201csuspicious\u201d in noninvasive encapsulated follicular variant of papillary thyroid cancers and should lead to ipsilateral lobectomy and isthmusectomy in the absence of historical, clinical, neck ultrasound or fine needle aspiration biopsy findings elsewhere suggesting metastatic lymph node involvement or synchronous thyroid malignancy ."} +{"text": "One strong possibility is that oxidative damage to the non-prion form of a protein may be an important trigger influencing the formation of its heritable prion conformation. We have examined the oxidative stress-induced formation of the yeast [PSI+] prion, which is the altered conformation of the Sup35 translation termination factor. We used tandem affinity purification (TAP) and mass spectrometry to identify the proteins which associate with Sup35 in a tsa1 tsa2 antioxidant mutant to address the mechanism by which Sup35 forms the [PSI+] prion during oxidative stress conditions. This analysis identified several components of the cortical actin cytoskeleton including the Abp1 actin nucleation promoting factor, and we show that deletion of the ABP1 gene abrogates oxidant-induced [PSI+] prion formation. The frequency of spontaneous [PSI+] prion formation can be increased by overexpression of Sup35 since the excess Sup35 increases the probability of forming prion seeds. In contrast to oxidant-induced [PSI+] prion formation, overexpression-induced [PSI+] prion formation was only modestly affected in an abp1 mutant. Furthermore, treating yeast cells with latrunculin A to disrupt the formation of actin cables and patches abrogated oxidant-induced, but not overexpression-induced [PSI+] prion formation, suggesting a mechanistic difference in prion formation. [PIN+], the prion form of Rnq1, localizes to the IPOD (insoluble protein deposit) and is thought to influence the aggregation of other proteins. We show Sup35 becomes oxidized and aggregates during oxidative stress conditions, but does not co-localize with Rnq1 in an abp1 mutant which may account for the reduced frequency of [PSI+] prion formation.Mammalian and fungal prions arise Prions are infectious agents which are composed of misfolded proteins and have been implicated in progressive neurodegenerative diseases such as Creutzfeldt Jakob Disease (CJD). Most prion diseases occur sporadically and are then propagated in a protein-only mechanism via induced protein misfolding. Little is currently known regarding how normally soluble proteins spontaneously form their prion forms. Previous studies have implicated oxidative damage of the non-prion form of some proteins as an important trigger for the formation of their heritable prion conformation. Using a yeast prion model we found that the cortical actin cytoskeleton is required for the transition of an oxidized protein to its heritable infectious conformation. In mutants which disrupt the cortical actin cytoskeleton, the oxidized protein aggregates, but does not localize to its normal amyloid deposition site, termed the IPOD. The IPOD serves as a site where prion proteins undergo fragmentation and seeding and we show that preventing actin-mediated localization to this site prevents both spontaneous and oxidant-induced prion formation. Sc) underlies the development of prion diseases in a mechanism which involves conversion of the normal prion protein (PrP) into its infectious PrPSc conformation prion [One strong possibility underlying the ormation . For exation [Sc \u201330 whileormation . Methionormation \u201334 and aormation . Oxidatiormation . The de +] prion \u201338. Prev+] prion , 38. Hende novo appearance of the [PSI+] prion is increased by overexpression of Sup35 in [PIN+][psi-] strains which increases the probability of forming prion seeds [de novo [PSI+] formation. This is particularly interesting given the increasing evidence suggesting that cytoskeletal structures provide a scaffold for the generation of protein aggregates. Insoluble aggregates of amyloid-forming proteins including prions are targeted to the IPOD as part of the cells\u2019 protein quality control system prion. Our data suggest a key role for the cortical actin cytoskeleton since we identified a number of components of the Arp2/3 actin-nucleation complex which specifically associate with Sup35 in the antioxidant mutant. We show that loss of several of these factors abrogates the increased frequency of [PSI+] prion formation which is normally observed in response to oxidative stress conditions. However, these mutants do not affect the increased frequency of [PSI+] prion formation induced in response to Sup35 overexpression. We show that Sup35 oxidative damage and aggregation occurs in actin-nucleation complex mutants in response to oxidative stress conditions, but the aggregates do not appear to form normally at the IPOD. Our data suggest that the cortical actin cytoskeleton is important for the formation of a propagating [PSI+] conformer following oxidant-induced misfolding and aggregation of Sup35.Oxidative stress provides a powerful tool to examine the PSI+] prion during oxidative stress conditions, we used tandem affinity purification (TAP) and mass spectrometry to identify proteins which associate with Sup35 in a tsa1 tsa2 mutant. For this analysis, we used [PIN+][psi-] versions of wild-type and tsa1 tsa2 mutant strains containing genomically-tagged Sup35. We have previously confirmed that TAP-tagging Sup35 does not affect reversible [PSI+] prion formation prion formation was quantified by analysing the formation of Ade+ colonies which arise due to nonsense suppression of the ade1-14 mutant allele. [PSI+]-mediated suppression can be differentiated from nuclear-encoded nonsense suppressor mutations by their elimination in guanidine hydrochloride (GdnHCl). The control [PIN+][psi-] strain was grown in the presence of 100 \u03bcM hydrogen peroxide for 20 hours prior to scoring [PSI+] prion formation. This oxidative stress treatment increased the frequency of [PSI+] prion formation by approximately ten-fold of [PSI+] and so we reasoned that we could use this concentration of LTA to test whether it affects the induction of [PSI+]. Rhodamine-phalloidin staining was used to visualize the cortical actin cytoskeleton and to confirm that the 10 \u03bcM LTA treatment disrupted the formation of actin patches (Treatment of yeast cells with latrunculin A (LTA) disrupts the formation of actin cables and patches . This hast cells . This ef patches .PIN+][psi-]-strain was grown in the presence of 10 \u03bcM LTA and 100 \u03bcM hydrogen peroxide for 20 hours to induce prion formation. We first examined Sup35 puncta formation by expressing SUP35NM-GFP for the final two hours of the oxidant treatment. Approximately 8% of wild-type cells contained visible Sup35 aggregates following exposure to hydrogen peroxide. This frequency was somewhat reduced in cells treated with 10 \u03bcM LTA, where 3.9% of cells examined contained visible Sup35 aggregates which was expressed under the control of the GAL1 galactose-regulated promoter form .PSI+] formation, but also reduced the spontaneous frequency of de novo [PSI+] formation was used for all experiments. Strains deleted for TSA1 (tsa1::LEU2) and TSA2 (tsa2::kanMX) and containing Sup35 tagged at its C-terminus with a tandem affinity purification (TAP) tag have been described previously expressing the Sup35NM domain conjugated to GFP under the control of the CUP1 promoter has been described previously [GAL1-SUP35NM-RFP [LEU2] [GAL1-RNQ1-EGFP [URA3] which expresses Rnq1-GFP under the control of the GAL1 promoter [UBC9 fused to GFP (GFP\u2013Ubc9ts) was expressed under the control of the GAL1 galactose-regulated promoter [The yeast plasmid eviously as has tP [LEU2] . Rnq1 wapromoter . The yeapromoter . A thermpromoter PSI+] prion formation. Cells were treated with 10 \u03bcM latrunculin to disrupt the actin cytoskeleton.Strains were grown at 30\u00b0C with shaking at 180 rpm in rich YEPD medium or minimal SD supplemented with appropriate amino acids and bases. SRaf media contained 2% w/v raffinose and SGal media contained 2% w/v galactose. Media were solidified by the addition of 2% (w/v) agar. Strains were cured by five rounds of growth on YEPD agar plates containing 4 mM guanidine hydrochloride (GdnHCl). Where indicated, strains were grown in the presence of 100 \u03bcM hydrogen peroxide for 20 hours prior to analysing [PSI+] prion formation was scored by growth in the absence of adenine. Diluted cell cultures were plated onto SD plates lacking adenine (SD-Ade) and incubated for 7\u201310 days. Colonies which grew on SD-Ade plates were counted and then picked onto new SD-Ade plates before replica-printing onto SD-Ade and SD-Ade containing 4mM GdnHCl. Colonies that grew on SD-Ade, but not on SD-Ade with GdnHCl were scored as [PSI+]. [PSI+] colonies were also scored by visual differentiation of red/white colony formation on YEPD plates and by the conversion of pink/white [PSI+] colonies to red [psi-] colonies on YEPD plates containing GdnHCl. For oxidant induced prion assays, cultures were grown in the presence of 100 \u03bcm hydrogen peroxide for 20 hours prior to scoring [PSI+] formation. For Sup35 overexpression-induced prion assays, cultures were grown in the presence of 50 \u03bcM copper sulphate for 20 hours to induce CUP1-SUP35NM-GFP expression prior to scoring [PSI+] formation [De novo [PIN+] formation was performed as previously described [pin\u2013] [psi\u2013] strains in order to detect cells that generate [PSI+] de novo. Since [PSI+] formation is dependent on cells being [PIN+] [PIN+] formation was estimated based on the number of [PSI+] cells which arise. [PSI+] and [PIN+] formation was calculated based on the means of at least three independent biological repeat experiments.The frequency of spontaneous [ormation . De novoescribed . Brieflyg [PIN+] , the ratCUP1-SUP35NM-GFP following 50 \u03bcM copper sulphate addition to induce the CUP1 promoter [psi-] versions of the wild-type, abp1, crn1 and pan1 mutant strains containing the Sup35NM-GFP plasmid induced with copper for 24 hours. Rhodamine-phalloidin staining was used to visualize the cortical actin cytoskeleton.Fluorescence micrographs are shown for [(TIF)Click here for additional data file.S1 Tabletsa1 tsa2 mutant strains and the associated proteins identified from three repeat experiments using mass spectrometry. This resulted in the identification of 63 and 47 proteins which are shown according to whether they were identified in the wild-type, tsa1 tsa2 mutant or both strains.Sup35-TAP was immunoprecipitated from the wild-type and (DOCX)Click here for additional data file.S2 TableThe proteins identified within each MIPS category classification shown in (XLSX)Click here for additional data file."} +{"text": "Bordetella associate with various animal hosts, frequently causing respiratory disease. Bordetella pertussis is the primary agent of whooping cough and other Bordetella species can cause similar cough illness. Here, we report four complete genome sequences from isolates of different Bordetella species recovered from human respiratory infections.Species of the genus Bordetella species are associated with a variety of hosts, where they are often the etiologic agents of disease. Most notably, Bordetella pertussis causes whooping cough (pertussis) but related species B.\u00a0parapertussis and B.\u00a0holmesii can cause similar pertussis-like illness in humans , B. bronchiseptica (I328), B.\u00a0holmesii (H903), and unclassified Bordetella sp. (H567) which were all recovered from patients with respiratory infection.n humans \u20135. B.\u00a0brry tract \u201310. ThroDe novo genome assembly of filtered reads was performed using the Hierarchical Genome Assembly Process (gidA), consistent with available genome sequences of Bordetella species. To ensure accuracy, assemblies were confirmed by comparison to BamHI and KpnI restriction digest optical maps using the Argus system (OpGen) with MapSolver and further \u201cpolished\u201d by mapping Illumina HiSeq PE-100 or MiSeq PE-300 reads using CLC Genomics Workbench . Final assemblies were annotated using the NCBI automated Prokaryotic Genome Annotation Pipeline (PGAP).Whole-genome shotgun sequencing was performed using a combination of the PacBio RSII , Illumina HiSeq/MiSeq , and Argus platforms as described previously . Brieflyciences) with at Bordetella and did not match any named species or group according to either 16S (nrdA (Bordetella species isolated from respiratory samples of cystic fibrosis patients (Achromobacter. Isolate H567 also behaved more similarly to species proposed by Vandamme et al. (Isolate and assembly characteristics are summarized in ther 16S , 14 or n6S (nrdA gene seqBordetella species have been recovered from human respiratory tract specimens, including putative novel species like isolate H567. The availability of complete genome sequences from species other than B.\u00a0pertussis should aid research of how the broader Bordetella genus causes respiratory illness in humans.Several The complete genome sequences have been deposited at DDBJ/EMBL/GenBank under the accession numbers listed in"} +{"text": "Additionally, 32 suggestive loci (p < 5x10-6) were observed. Several candidate genes were located in these loci, such as NLK, MEF2A, SOX9 and SOX11. Genome-wide linkage analysis of cranial vault shape in mice (N = 433) was performed to follow-up the associated candidate loci identified in the human GWAS. Two loci, 17q11.2 (c11.loc44 in mice) and 17q25.1 (c11.loc74 in mice), associated with cranial vault size in humans, were also linked with cranial vault size in mice (LOD scores: 3.37 and 3.79 respectively). These results provide further insight into genetic pathways and mechanisms underlying normal variation in human craniofacial morphology.The shape of the cranial vault, a region comprising interlocking flat bones surrounding the cerebral cortex, varies considerably in humans. Strongly influenced by brain size and shape, cranial vault morphology has both clinical and evolutionary relevance. However, little is known about the genetic basis of normal vault shape in humans. We performed a genome-wide association study (GWAS) on three vault measures in a sample of 4419 healthy individuals of European ancestry. All measures were adjusted by sex, age, and body size, then tested for association with genetic variants spanning the genome. GWAS results for the two cohorts were combined via meta-analysis. Significant associations were observed at two loci: 15p11.2 (lead SNP rs2924767, p = 2.107 \u00d7 10 FGFR1 gene, chosen because mutations in this gene have been implicated in craniosynostosis syndromes. While these studies were promising, a large-scale genome-wide investigation of normal cranial vault morphology had been lacking.The dimensions of the human cranial vault, which forms the protective skeletal covering around the brain , are higTo address this deficit, we examined the genetic basis of cranial vault morphology in 4419 individuals of recent European ancestry by performing genome-wide association studies (GWASs) of maximum cranial width (MCW), maximum cranial length (MCL), and the cephalic index (CI) in two independent cohorts. Subsequently, genome-wide significant and suggestive loci were tested using linkage analysis in a sample of mice with comparable cranial vault measurements available.-8 was set for genome-wide statistical significance.GWAS was performed for MCW, MCL and CI with 968515 (for the 3DFN cohort) or 567677 (for the OFC cohort) genotyped and imputed SNPs with MAF > 1% in two separate European-derived cohorts using either linear regression or mixed models. GWAS results for each cohort were then combined via inverse variance-weighted meta-analysis using Stouffer\u2019s p-value method . To. To27]. For the 3DFN cohort, DNA was extracted from saliva samples and genotyped along with 72 HapMap control samples for 964193 SNPs on the Illumina HumanOmniExpress+Exome v1.2 array plus 4322 SNPs of custom content by the Center for Inherited Disease Research (CIDR). For the OFC cohort, DNA was extracted from saliva, blood, or other biological samples and genotyped along with the same 72 HapMap controls for 551787 SNPs on an Illumina HumanCore+Exome array plus 15890 SNPs of custom content, also by CIDR. Genetic data cleaning and quality control analyses were performed as described in detail, previously ,29. In bTo assess population structure, we performed principal component analysis (PCA) within each cohort using subsets of uncorrelated SNPs. Based on the scatterplots of the principal components (PCs) and scree plots of the eigenvalues , we deteA total of 9482681 and 9211574 imputed SNPs were tested for the 3DFN and OFC cohorts respectively. Imputation of unobserved variants was performed using haplotypes from the 1000 Genomes Project Phase 3 as the reference. Imputation was performed using IMPUTE2 . We used-8 was set for genome-wide statistical significance. QQ plots and genomic inflation scores are represented in Genetic association with MCW, MCL and CI was tested in 3DFN and OFC cohorts separately for 968515 (for the 3DFN cohort) or 567677 (for the OFC cohort) genotyped and imputed SNPs with MAF > 1%. For 3DFN, GWAS was performed using linear regression while adjusting for sex, age, height, weight and four PCs of ancestry as implemented in PLINK . For OFC2 > 0.8) with the lead SNPs. Genes of interest were defined based on physical proximity of 500 kb to the lead SNP at each locus. These genes were queried in the following online databases: The Mouse Genome Informatics (MGI) database . Cranial measurements for the OFC cohort are also available through the same dbGaP accession. Cranial measurements for the 3D Facial Norms cohort are available through the FaceBase Consortium . A full description of the 3D Facial Norms dataset is available at https://www.facebase.org/facial_norms/. Although there are no costs associated with access to FaceBase datasets, users must formally apply for access to human datasets through the FaceBase Consortium (the application process is described here: https://www.facebase.org/methods/policies/). Full summary statistics for all SNPs are available upon request.The genotype data for both human cohorts are available through dbGaP [S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S7 Table(XLSX)Click here for additional data file.S8 Table(DOCX)Click here for additional data file.S1 Fig-8.The horizontal line represents the conventional threshold for genome-wide statistical significance: p \u2264 5x10(PDF)Click here for additional data file.S2 FigLocusZoom plots show the association with facial traits. Genotyped SNPs are depicted by asterisks and imputed SNPs are depicted by circles. Shading of the points represent the linkage disequilibrium between each SNP and the top SNP, indicated by purple shading. Grey points in these plots represent the lack of LD information between the index SNP (diamond) the plotted SNP (circle or asterisk). The blue overlay shows the recombination rate (right y-axis). Positions of genes are shown below the plot.(PDF)Click here for additional data file.S3 FigLocusZoom plots show the association with facial traits. Genotyped SNPs are depicted by asterisks and imputed SNPs are depicted by circles. Shading of the points represent the linkage disequilibrium between each SNP and the top SNP, indicated by purple shading. Grey points in these plots represent the lack of LD information between the index SNP (diamond) the plotted SNP (circle or asterisk). The blue overlay shows the recombination rate (right y-axis). Positions of genes are shown below the plot.(PDF)Click here for additional data file.S4 FigLocusZoom plots show the association with facial traits. Genotyped SNPs are depicted by asterisks and imputed SNPs are depicted by circles. Shading of the points represent the linkage disequilibrium between each SNP and the top SNP, indicated by purple shading. Grey points in these plots represent the lack of LD information between the index SNP (diamond) the plotted SNP (circle or asterisk). The blue overlay shows the recombination rate (right y-axis). Positions of genes are shown below the plot.(PDF)Click here for additional data file.S5 FigThe three traits are indicated by color: MCW = Orange, MCL = Green, CI = Black. The horizontal line represents the permutation-based empirical threshold for genome-wide statistical significance.(PDF)Click here for additional data file.S6 Fig(PDF)Click here for additional data file.S7 Fig(TIFF)Click here for additional data file.S8 Fig(PDF)Click here for additional data file."} +{"text": "The mutant developed normal green leaves from leaf 5, but reduced tillering and chlorotic leaves and panicles appeared later. Chlorotic yss2 seedlings have decreased pigment contents and impaired chloroplast development. Genetic analysis showed that the mutant phenotype was due to a single recessive gene. Positional cloning and sequence analysis identified a single nucleotide substitution in YSS2 gene causing an amino acid change from Gly to Asp. The YSS2 allele encodes a NDPK2 (nucleoside diphosphate kinase 2) protein showing high similarity to other types of NDPKs. Real-time RT-PCR analysis demonstrated that YSS2 transcripts accumulated highly in L4 sections at the early leaf development stage. Expression levels of genes associated with Chl biosynthesis and photosynthesis in yss2 were mostly decreased, but genes involved in chloroplast biogenesis were up-regulated compared to the wild type. The YSS2 protein was associated with punctate structures in the chloroplasts of rice protoplasts. Our overall data suggest that YSS2 has important roles in chloroplast biogenesis.Chloroplast development and chlorophyll (Chl) biosynthesis in plants are regulated by many genes, but the underlying molecular mechanisms remain largely elusive. We isolated a rice mutant named FtsZ (encoding a component of the plastid division machinery) is required for the first step , rbcL and psbA (encoding the D1 subunit of the PSII complex) are abundant in the third step, which functions in activation of the photosynthetic apparatus and OsWP1 play essential roles in chloroplast development of leaf and panicle. Weak allele mutation in WLP1 caused albino seedling phenotype and bleached panicles, especially severe under lower temperature. Chloroplast development could be affected in the seedlings and young panicles of wlp1 mutant, although data are not available or a virescent phenotype (wp1 showed striated leaf in seedlings and white panicles at heading). The wp1 mutant arrested plastidic protein synthesis and biogenesis of chloroplast ribosomes and was defective in early chloroplast development with high similarity to NDPKs in other species. Expression analysis showed that YSS2 was highly expressed in L4 tissues, the key time of chloroplast biogenesis. Subcellular localization showed that YSS2 is a chloroplast-associated protein. These results implied that YSS2 plays important roles in chloroplast biogenesis.In this study, we aimed to characterize a young seedling stripe mutant, yss2 was identified in an MNU-mutagenized population of japonica cultivar Nongyuan 238. Plants were grown in a growth chamber or paddy fields. Crosses between the yss2 mutant and Nongyuan 238 or Nanjing 11 were separately used for genetic analysis and gene mapping. Seeds of cultivars Nongyuan 238 and Nanjing 11 were obtained from the Chinese National Key Facility for Crop Gene Resources and Genetic Improvement in Beijing.The white leaf and panicle mutant Chls and Car were assayed spectrophotometrically according to methods described previously and 93-11 (indica). Primer pairs designed with Primer Premier 5.0 are listed in yss2 mutant were amplified and sequenced.Following initial mapping of the yss2 seedlings using an RNA Prep Pure Plant kit (Tiangen) and reverse-transcribed using a SuperScript II kit (TaKaRa). Real-time RT-PCR was performed using a SYBR\u00ae Premix Ex TaqTM kit (TaKaRa) on a LightCycler 480 Real-Time PCR System (Roche). The 2-\u0394\u0394CT method was used to analyze relative gene expression (https://www.genscript.com/ssl-bin/app/primer). The ubiquitin gene (LOC_Os03g13170) (Ubq) was used as a reference control.Total RNA was extracted from wild-type and pression . Primershttp://rice.plantbiology.msu.edu/cgi-bin/gbrowse/rice/). Homologous sequences of YSS2 were identified using the blastp search mode at NCBI (http://www.ncbi.nlm.nih.gov/) and sequences were aligned with BioEdit software. A neighbor-joining tree based on 1,000 bootstrap replicates was generated using MEGA v4.1 software. Expression profiles of YSS2, OsNDPK1 (LOC_Os07g30970) and OsNDPK3 (LOC_Os05g51700) were obtained from the RiceXPro database (http://ricexpro.dna.affrc.go.jp/). Subcellular localization of YSS2 was predicted using the ChloroP (Candidate genes were predicted by the RGAP database (YSS2 was amplified and cloned to the N-terminus of GFP in the transient expression vector pA7-GFP (primer pairs shown in The coding sequence of yss2 mutant was originated from an N-methyl-N-nitrosourea (MNU) mutagenized population of japonica cultivar (cv.) Nongyuan 238. The mutant seedlings displayed a striated leaf phenotype in seedling leaves 2 to 4 under paddy field conditions of yss2 leaves had well-developed lamellar structures with normally stacked grana and thylakoid membranes similar to wild type plants (yss2 mutant carried a single nucleotide change (G to A) in the second exon relative to wild type, resulting in an amino acid change from Gly to Asp at position 83 , Table 3ition 83 . To veriYSS2 allele with seven exons and six introns encodes a polypeptide of 220 amino acid residues with a predicted molecular mass of 23.5 kDa , HEMA1 (encoding glutamyl tRNA reductase), YGL1 (encoding Chl synthetase), CHLI and CHLD (encoding Mg-chelatase I and D subunits) were clearly decreased in yss2 seedlings compared to wild type. However, there was no difference in expression of CHLH (encoding Mg-chelatase H subunit) , psbA, psbB and psbC (encoding PSII subunits), LHCP (encoding PSII-associated light-harvesting chlorophyll protein) and psaB (encoding PSI subunit) was distinctly down-regulated in the mutant and second steps of chloroplast biosynthesis were up-regulated in the yss2 mutant compared to wild type (YSS2 is involved in the regulatory network of Chl biosynthesis and photosynthesis as well as chloroplast formation.Given the phenotypic difference between the subunit) . Expresse mutant . Genes rild type . These dTargetP , whereasysa, ylc1, yss1 and wsl (ygl1 and vyl) display the mutant phenotype and delayed plant growth throughout the entire life cycle and photosynthetic genes were separately up- and down-regulated in yss2 mutant (yss2 might decrease PEP activity and suppress plastid transcription. The dramatically elevated levels of rpo genes and decreased expression of photosynthetic genes indicated that the mutants lacked chloroplast ribosomes or reduced plastid DNA contents (YSS2 might have important roles in the regulatory network of both. Nevertheless, the reduced expression of genes for Chl biosynthesis is not strong and could be an indirect effect such as retrograde plastid-to-nucleus signaling that disturbs the expression of nuclear-encoded chloroplast genes (e region . Genomicition 83 . YSS2 wa protein . These d tissues . Althoug tissues , suggest2 mutant , suggestcontents , implyinV2, was found to participate in chloroplast biogenesis (YSS2 is involved in chloroplast biogenesis and that the yss2 mutation disrupts the metabolic balance between NTPs and NDPs, a consequence of which is impaired photosynthesis and hindered plant growth. Sequence alignment and phylogenetic analysis revealed that YSS2 is an evolutionarily conserved protein with a NDPK domain. The amino acid Gly in the NDPK domain is highly conserved in all NDPK proteins (It has been reported that NDPKs regulate the metabolic balance between NTPs and NDPs by catalyzing the transfer of phosphate groups and are involved in cell growth and division, embryo and seed development, signal transduction and plant stress response (proteins and 4. I"} +{"text": "Aythyafuligula) and Eurasian wigeons (Anas penelope).Recurrence of avian influenza outbreaks might alter wild bird populationdynamics.During autumn\u2013winter 2016\u20132017, highly pathogenic avian influenzaA(H5N8) viruses caused mass die-offs among wild birds in the Netherlands. Amongthe \u224813,600 birds reported dead, most were tufted ducks ( Unidentified waterbird carcasses probably also mostly represented thesespecies. H5N8 infection was confirmed in 21 species and not detected among the lownumbers of sampled birds representing 13 other species and Eurasian coot(Fulica atra [4%]), in addition to great crested grebe(Podiceps cristatus), mute swan, greater scaup (Aythyamarila), and several goose and gull species (each <1%).After the first H5N8 detection in diseased waterbirds on November 8, hundreds ofcarcasses were found at Gouwzee and Wolderwijd.Deaths at these locations peaked within 10 days, with \u22485,300 carcassesreported by November 18 . AnestiBeginning in late November, outbreak hotspots moved from open water to water-richagricultural areas ; Video. Because these data are based on numbers of reported carcasses, they provide anunderestimation of actual deaths. Although carcass detection rates during dailysearches at Gouwzee and Wolderwijd were estimated to be 90%\u201395% , search efficiency was probably much lower at otheroutbreak hotspots. Collection rates of waterbird carcasses during typical avianbotulism outbreaks are 10%\u201325% yielded similar results to compare the number of deaths per species group duringNovember 2016\u2013January 2017 with those occurring in the same timeframe from2010\u20132011 to 2015\u20132016 . Death c results Figure.Larus marinus) and 11%\u201339% of thewintering population of peregrine falcons weresimilarly affected. Stronger effects were observed locally. At Gouwzee,\u22486,000 tufted ducks were counted in December after \u22482,000 of them haddied in November. Assuming that no migration occurred, we estimate that up to 25% ofthe local population of tufted ducks might have died, which might affect populationdynamics substantially. Additional studies are needed to evaluate long-term impactson these populations and to elucidate why high numbers of birds survived or escapedinfection.The elevated number of deaths among wild birds raises concern about potentialpopulation effects. After accounting for detection probability . Awareness of clinical signs in wild birds mightfaReadily available specific guidelines would help management of HPAI virus outbreaksin wild birds. National HPAI preparedness plans should include specific protocolsabout how to handle carcasses and whatto report . Moreover, sufficient resources should be available for adequate samplingand testing of specimens to rule out other diseases and to track virus dynamicsduring an outbreak.,Our findings indicate that the 2016\u20132017 H5N8 outbreaks in the Netherlandswere associated with unprecedented high HPAI-related mortality rates in a wide rangeof wild bird species. These latest H5N8 outbreaks have shifted the paradigm of wildbirds as unaffected agents of HPAI viruses, with increasing concerns about potentialeffects on their populations. The Netherlands and other important staging areas formigratory waterbirds across Eurasia that have been affected by the 2016\u20132017H5N8 outbreaks (Overview of organizations and authorities that provided data for analysis ofwild bird deaths and a full species account of reported cases of aviandeaths during an outbreak of highly pathogenic avian influenza A virussubtype H5N8, the Netherlands, November 2016\u2013January 2017.Relative number of deaths among wild birds, based on data from the NatureInformation Foundation and an overview of clinical signs in wild birdsreported during an outbreak of highly pathogenic avian influenza A virussubtype H5N8, the Netherlands, November 2016\u2013January 2017."} +{"text": "Mc3r transcription in hypothalamic and limbic neurons improves appetitive responses during hypocaloric conditioning while having minor effects on nutrient partitioning, suggesting orexigenic functions. Rescuing hypothalamic MC3Rs also restores responses of fasting-responsive hypothalamic orexigenic neurons in hypocaloric conditions, suggesting actions that sensitize fasting-responsive neurons to signals from nutrient sensors. MC3R signaling in ventromedial hypothalamic SF1(+ve) neurons improves metabolic control, but does not restore appetitive responses or nutrient partitioning. In summary, desensitization of fasting-responsive orexigenic neurons may underlie attenuated appetitive responses of MC3R-deficient mice in hypocaloric situations. Further studies are needed to identify the specific location(s) of MC3Rs controlling appetitive responses and partitioning of nutrients between fat and lean tissues.Melanocortin neurons conserve body mass in hyper- or hypo-caloric conditions by conveying signals from nutrient sensors into areas of the brain governing appetite and metabolism. In mice, melanocortin-3 receptor (MC3R) deletion alters nutrient partitioning independently of hyperphagia, promoting accumulation of fat over muscle mass. Enhanced rhythms in insulin and insulin-responsive metabolic genes during hypocaloric feeding suggest partial insulin resistance and enhanced lipogenesis. However, exactly where and how MC3Rs affect metabolic control to alter nutrient partitioning is not known. The behavioral phenotypes exhibited by MC3R-deficient mice suggest a contextual role in appetite control. The impact of MC3R-deficiency on feeding behavior when food is freely available is minor. However, homeostatic responses to hypocaloric conditioning involving increased expression of appetite-stimulating (orexigenic) neuropeptides, binge-feeding, food anticipatory activity (FAA), entrainment to nutrient availability and enhanced feeding-related motivational responses are compromised with MC3R-deficiency. Rescuing Obesity is often attributed to a combination of genetic susceptibility and imbalances between energy intake and expenditure . These neurons integrate humoral cues of metabolic condition and neuropeptide Y (NPY) are activated upon fasting , ventral tegmental area (VTA), and medial habenula (MHb) mice on mixed or congenic (C57BL/6J) backgrounds has been inconclusive that reduce receptor binding and maximal cAMP accumulation in cell-based assays exhibit reduced musculoskeletal mass and increased adiposity when compared to mice inheriting \u201cwild type\u201d hMC3Rs (MC3RhWT/hWT) , over time this could produce significant changes in adiposity mutation on second messenger signaling thus far has been limited to measuring cAMP accumulation in the presence of the synthetic analog -\u03b1-MSH. Information on how the mutation alter other signaling mechanisms and responses to other ligands such as AgRP are not available, but could be relevant given that physiological responses to centrally administered melanocortin agonists involve distinct G protein signaling mechanisms when subjected to a hypocaloric diet presented at 24 h intervals are also attenuated ;MC3R(+ve) neurons in the hypothalamus are sufficient to restore \u201cnormal\u201d activity of NAG neurons. This could indicate a developmental role in which NAG neurons fail to develop normal responses to altered signals of metabolic state in the absence of MC3Rs. Alternatively, MC3Rs in the mature hypothalamus may exert an active \u201cgating\u201d function; determining whether rescuing MC3Rs in the adult mouse restores responses of NAG neurons to metabolic cues could address this question.We developed the LoxTBMc3r-deficient mice exhibiting lower dopamine and altered sucrose consumption and taste preferences neurons in the VTA improved motivational responses (+ve) and (\u2212ve) neurons in the VTA, with female Early experiments examining hypophyseal and autonomic outputs from the CNS controlling metabolism by melanocortins suggested no requirement for MC3R signaling. Acute stimulation of sympathetic activity by melanotan-II (MTII), an \u03b1\u2013MSH analog, requires functional MC4Rs are insulin resistant. While rhythms in insulin and glucose ad libitum fed Mc3r\u2212/\u2212 mice were normal, this result might be misleading. Fasting insulin, fasting glucose and glucose tolerance are normal in muscle-specific insulin receptor knockout mice (MIRKO) expressing neurons in the VMH . Analysis of body composition using a regression approach of MC3Rs affecting nutrient partitioning remains unclear.The functions of neural MC3Rs received little attention after the publication of the phenotypes of n Figure . In humaAB prepared the first manuscript draft. CG, MM, JT, HM, DM, and SF reviewed and edited the manuscript.Some of the work cited in the article was supported by grants from the National Institutes of Health (DK073189) to AB. AB also thanks the support of the Pennington Biomedical Research Foundation, Clinical Nutrition Center Grant P30 DK072476 , The Scripps Florida Fund and financial support from Saint Louis University.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Entirely DNA-free CRISPR-Cas9 systems using either Cas9 protein or Cas9 mRNA and chemically synthesized gRNAs allow for transient expression of CRISPR-Cas9 components, thereby reducing the potential for off-targeting, which is a significant advantage in therapeutic applications. In addition, the use of synthetic gRNA allows for the incorporation of chemical modifications for enhanced properties including improved stability. Previous studies have demonstrated the utility of chemically modified gRNAs, but have focused on one pattern with multiple modifications in co-electroporation with Cas9 mRNA or multiple modifications and patterns with Cas9 plasmid lipid co-transfections. Here we present gene editing results using a series of chemically modified synthetic sgRNA molecules and chemically modified crRNA:tracrRNA molecules in both electroporation and lipid transfection assessing indel formation and/or phenotypic gene knockout. We show that while modifications are required for co-electroporation with Cas9 mRNA, some modification patterns of the gRNA are toxic to cells compared to the unmodified gRNA and most modification patterns do not significantly improve gene editing efficiency. We also present modification patterns of the gRNA that can modestly improve Cas9 gene editing efficiency when co-transfected with Cas9 mRNA or Cas9 protein (> 1.5-fold difference). These results indicate that for certain applications, including those relevant to primary cells, the incorporation of some, but not all chemical modification patterns on synthetic crRNA:tracrRNA or sgRNA can be beneficial to CRISPR-Cas9 gene editing.Since its initial application in mammalian cells, CRISPR-Cas9 has rapidly become a preferred method for genome engineering experiments. The Cas9 nuclease is targeted to genomic DNA using guide RNAs (gRNA), either as the native dual RNA system consisting of a DNA-targeting CRISPR RNA (crRNA) and a Streptococcus pyogenes endonuclease Cas9 can bind DNA sequences upstream of an NGG protospacer adjacent motif (PAM) and cause a double-strand break (DSB). When a DSB occurs in mammalian cells, it is repaired by endogenous cellular mechanisms such as non-homologous end joining (NHEJ) or homology-directed repair (HDR). NHEJ is the predominant repair pathway and results in either perfect resolution of the DSB or imperfect repair with either insertions or deletions (indels) of nucleotides at the break site. The result of this imperfect repair can be an alteration of the downstream gene product, potentially causing a functional gene knockout.The class II CRISPR-Cas system is a bacterial adaptive defense mechanism that has been applied in mammalian cells for genome engineering \u20134. Usingtrans-activating crRNA (tracrRNA) was diluted in SuperBlock and incubated overnight at 4\u00b0C. Membranes were washed and secondary antibody was diluted 1:20,000 in SuperBlock (PBS formulation) with 0.5% Tween20 and incubated with membranes for 2 hours at room temperature. The membranes were washed and then submerged in SuperSignal\u2122 West Dura Substrate solution for beta-actin blots and Super West Femto Maximum Sensitivity Substrate for Cas9, and exposed to film.(TIF)Click here for additional data file.S3 FigPSMD7 (A.) or PSMD11 (B.), at multiple concentrations when transfected at 1.5625 nM to 50 nM at 2-fold increments into a stably expressing Cas9 U2OS cell line. Error bars are representative of biological triplicates. C. Average cell viability of unmodified or modified guide RNAs for two genes (PSMD7 and PSMD11) resulted in a significant decrease in cell viability (< 60%) for some modification patterns at concentrations higher than 6.25 nM. NTC = Non-targeting control. Error bars are representative of the average of biological triplicates of two genes with the same modification pattern in one experiment.Gene editing efficiency of unmodified (unmod) and modified crRNA:tracrRNA or sgRNA showed similar levels of gene editing efficiencies (< 1.5-fold difference) measured by EGFP fluorescence from knockout of a proteasome component, (TIF)Click here for additional data file."} +{"text": "WHO recommends antiretroviral treatment (ART) for all HIV-positive individuals. This study evaluated the association between baseline CD4 count and attrition in a cohort of HIV positive adults initiating ART at three department of health (DOH) clinics routinely providing ART at baseline CD4 counts >500cells/\u03bcL for the HPTN 071 (PopART) trial.All clients attending the DOH clinics were managed according to standard care guidelines with the exception that those starting ART outside of pertinent local guidelines signed research informed consent. DOH data on all HIV-positive adult clients recorded as having initiated ART between January 2014 and November 2015 at the three study clinics was analysed. Attrition, included clients lost to follow up or died, and was defined as \u2018being three or more months late for an antiretroviral pharmacy pick-up appointment\u2019. All clients were followed until attrition, transfer out or end May 2016.A total of 2423 clients with a median baseline CD4 count of 328 cells/\u03bcL (IQR 195\u2013468) were included of whom 631 (26.0%) experienced attrition and 140 (5.8%) were TFO. Attrition was highest during the first six months of ART . Higher attrition was found amongst those with baseline CD4 counts > 500 cells/\u03bcL compared to those with baseline CD4 counts of 0\u2013500 cells/\u03bcL This finding was confirmed on subset analyses when restricted to individuals non-pregnant at baseline and when restricted to individuals with follow up of > 12months.Attrition in this study was high, particularly during the first six months of treatment. Attrition was highest amongst clients starting ART at baseline CD4 counts > 500 cells/\u03bcL. Strategies to improve retention amongst ART clients, particularly those starting ART at baseline CD4 counts >500cells/\u03bcL, need strengthening. Improved monitoring of clients moving in and out of ART care and between clinics will assist in better understanding attrition and ART coverage in high burden countries. There are 36.7 million HIV positive individuals and 19.5 million people on antiretroviral treatment (ART) worldwide . To achiRetention in ART programmes in high burden settings is extremely challenging. A recent systematic review, which included 1.5 million participants from African and Asian programmatic studies (75% from Africa), the majority of whom started ART at baseline CD4 counts < 200cells/\u03bcL, found 17% and 26% of individuals on ART lost to follow up or died (attrition) at 12 months and at 24 months respectively . The medFollowing results of the START and TEMPRANO randomised control trials (RCTs) in 2015 , 11, WHOThis study was conducted at three DOH primary health care (PHC) clinics included in the \u2018Population effect of antiretroviral therapy to reduce HIV incidence\u2019 HPTN 071 (PopART) trial in the Western Cape (WC), South Africa. A full description of the HPTN 071 (PopART) trial design has been published . The comTwo clinics were located in the metro district (Metro 1 and 2) and one in a rural district . These study clinics offered ART regardless of CD4 count for all HIV positive clients aged 18 or older. During the study period for standard care at other clinics DOH ART guidelines recommended ART initiation at baseline CD4 count \u2264 350 cells/\u03bcL until January 2015 and thereafter at baseline CD4 count \u2264500cells/\u03bcL [A fixed-dose combination of tenofovir, emtricitabine and efavirenz (TEE) was used for first line treatment. Pharmacy pick up dates for collection of TEE were initially scheduled monthly, then every two to three months once clients were assessed as stable on ART by a clinician. CD4 count was routinely measured at four months and 12 months of ART and viral load at 4 months, 12 months and then annually . All rouThis study included data on all clients 18 years and older recorded in Tier.net as initiating ART at the three study clinics, between 1 January 2014 and 30 November 2015 . Follow TM. CD4 count data, extracted from the NHLS database, were linked to data in Tier.net using the WC DOH \u2018Clinicom number\u2019 as unique clinic identifier in Stata13TM. Pharmacy pick up data recorded in Tier.net was used to calculate the date for next scheduled clinic appointment. Data cleaning and validation included cross-referencing data fields within Tier.net and across Tier.net, NHLS and ETR.net databases. Data elements in Tier.net that were adjudged to have with high rates of missing or incorrect data, e.g. baseline WHO stage and data on adherence club attendance were excluded from analysis.All data were initially extracted from Tier.net except the data on baseline TB treatment which were extracted from the electronic TB register (ETR.net). If the baseline CD4 count results were missing from Tier.net, these were extracted from the National Health Laboratory Services (NHLS) databases. Data from Tier.net were linked to ETR.net data through a matching algorithm utilising name, surname and date of birth in Microsoft SQL Serverth May 2016 (end of the study); whichever was the earliest. Unadjusted and adjusted comparisons of the hazard of attrition at different baseline CD4 count strata were carried out using Cox regression. Potential confounding baseline characteristics for inclusion in the adjusted analysis were selected a priori based on clinical relevance; these included: age, sex, pregnancy status, TB treatment, clinic, previous ART exposure of more than 3 months and year of ART start. Baseline CD4 count strata were chosen to align with previous ART guideline cut offs. [Baseline characteristics were described for continuous and categorical variables and distribution across CD4 strata was assessed using Chi Squared tests and Kruskal-Wallis tests. Incidence rates were estimated and time-to-event analyses were conducted using Kaplan Meier survival and smoothed hazard estimates. Clients were censored on either the date of attrition, TFO, or on 30ut offs. ProportiThe HPTN 071 (PopART) trial was approved by the Stellenbosch University Health Research Ethics Committee (SU HREC) (Ref. No. N12/11/074) and the London School of Hygiene and Tropical Medicine Research Ethics Committee (Reference number 6362). All clients initiating ART outside local DOH guidelines for HPTN 071 (PopART) gave written informed consent. Further permission for this study and the use of individual level data from the WC DOH sources with a waiver for informed consent has also been received from SU HREC (reference number N12/11/074A), the Western Cape Government (Reference no. WC_2015RP51_715) and City of Cape Town (Reference no. 10529).A total of 2593 clients who started ART at the study clinics between 1 January 2014 and end November 2015 were screened for inclusion in the study, of whom 170 (6.6%) were excluded due to missing baseline CD4 counts. This left a sample of 2423 clients included in the analysis. The distribution of clients by baseline CD4 count strata was 631 (26.0%) at CD4 0\u2013200 cells/\u03bcL, 708 (29.2%) at CD4 201\u2013350 cells/\u03bcL, 582 (24.0%) at CD4 351\u2013500 cells/\u03bcL and 502 (20.7%) at CD4 >500 cells/\u03bcL. Overall, 631 (26.0%) clients experienced attrition during 2389 person years (PY) of follow up (Incidence Rate (IR): 26.4/100 PY) and 140 (5.8%) were TFO. Amongst individuals experiencing attrition, 11 (1.7%) were documented in Tier.net as having died. Median baseline CD4 count amongst those individuals who died was 34 cells/\u03bcL (IQR: 63\u2013155). Median follow up time was 11.2 (IQR 7.2\u201316.1) months. Cumulative numbers of clients experiencing attrition was 418 (17.3%), 561 (23.2%), 613 (25.3%), 631 (26.0%) at 6, 12, 18 and 24 months on ART respectively). Kaplan Meier estimates showed higher attrition amongst clients with baseline CD4 counts > 500cells/\u03bcL compared to \u2264 500cells/\u03bcL (P<0.02). Adjusted Cox regression analysis using the full model showed higher attrition amongst clients with baseline CD4 counts > 500 cells/\u03bcL compared to 0-500cells/\u03bcL. Additional multivariate analysis that further stratified baseline CD4 count showed higher attrition in individuals with baseline CD4>500cells/\u03bcL when compared to those with baseline CD4 counts 350\u2013500 cells/\u03bcL . Multivariate logistic regression of factors associated with missing baseline CD4 counts showed increased rates of missing baseline CD4 counts amongst clients who were pregnant at baseline and amongst those starting ART in 2014 compared to 2015. Being treated at Metro 2 and the rural clinic were associated with decreased rates of missing baseline CD4 counts.In this study we found, in a cohort of clients receiving ART regardless of CD4 count at three DOH clinics, we found high rates of attrition, higher amongst clients who started ART with baseline CD4 counts > 500cells/\u03bcL compared to those with baseline CD4 count 0\u2013500 cells/\u03bcL. This finding was confirmed in subset analyses restricted to individuals non- pregnant at baseline and when restricted to individuals with follow up time of > 12 months. When dividing baseline CD4 count to four strata, attrition was higher amongst clients with baseline CD4 counts > 500 cells/\u03bcL compared to those with baseline CD4 counts of 350\u2013500 cells/\u03bcL.The cumulative proportion of clients experiencing attrition in this paper was comparable to that reported by DOH for the corresponding health districts during the same time period ; as wellThis finding of higher attrition amongst individuals with higher baseline CD4 counts, who may be more likely to be clinically well when starting ART, supports findings from previous studies. \u20139. The uThe extent to which mortality contributed to attrition was not accurately documented in Tier.net. Although mortality as a cause of attrition is decreasing in Africa , it remaThe peak in attrition during the first six months on ART across all CD4 strata emphasises the need for additional retention strategies at clinics during early ART. , 25\u201327ThDespite a well-established ART service in the Western Cape since 2004 and provision of ART regardless of CD4 count for HPTN 071 (PopART) since January 2014 a high proportion of clients (26.0%) in this study initiated ART at baseline CD4 counts < 200 cells/\u03bcL. This persistence of low baseline CD4, even in the context of ART regardless of CD4 count, is a serious concern. Interventions aimed at promoting earlier ART uptake should therefore continue to be a priority and further strengthened. A recent systematic review of community and clinic based interventions aimed at increasing uptake of ART in sub-Saharan Africa found home based HIV testing and improved efficiencies and structure at clinics to be effective in improving ART uptake [The study has a number of key strengths. Data used in the analysis was part of a high quality routine dataset, strengthened by prospective data quality improvement for HPTN 071 (PopART) and clients were provided ART regardless of CD4 count ahead of recent changes to WHO and local guidelines. There were high rates of completeness in key data fields with only a small proportion (6.6%) of eligible clients excluded from the analysis due to missing baseline CD4 count results. There was also high similarity in baseline characteristics associated with attrition between clients excluded due to missing baseline CD4 counts and those included in the study. The three study clinics were typical of metro and rural clinics in the Western Cape and clinic activities were closely aligned to standard care during the study period, supporting generalisability of study findings. Another major strength was the use of an objective measure for determination of attrition (pharmacy pick up date).There are, however, a number of limitations to consider. Factors not measured by the available date set including psychosocial and health systems factors may have confounded the association between baseline CD4 count and attrition. In this regard; although activities at clinics included in this study were closely aligned for PopART and choice of clinic was not associated with attrition on multivariate analysis it is possible that clinic-related factors not measured in this paper may have confounded the primary analysis. Data and resources required for active follow up of individuals experiencing attrition to determine whether they had died or transferred to another facility without informing their current facility (silent transfers) were not available for this study. Published data show high rates of silent transfer to other ART clinics amongst individuals documented lost to follow up at PHC clinics in the Western Cape. The exteThroughout HPTN 071 (PopART) additional support was provided through staff and health systems support at clinics and in the community by CHiPs workers. This support was likely to have reduced overall attrition. Although all ART clients were meant to have received community-based support from both DOH CCWs and/or CHiPs teams, it was not known what proportion of clients received community-based support or whether the community based support differentially impacted clients with baseline CD4 counts > 500cells/\u03bcL.There are further limitations affecting the generalisability of these results. Clients starting ART outside of pertinent DOH guidelines received additional counselling during the signing of research informed consent and which may have in turn reduced their risk of attrition amongst individuals with baseline CD4 >500cells/\u03bcL.We documented higher attrition amongst clients initiating ART at baseline CD4 counts > 500cells/\u03bcL, highlighting an urgent need for retention with a focus on clients initiating ART at higher baseline CD4 counts. At the same time, strategies to improve earlier uptake of ART before their CD4 counts fall below 200cell/\u03bcL need to be strengthened. Monitoring systems that more accurately measure the contribution of death and silent transfers to attrition from ART programmes will assist in a better understanding of retention in ART programmes and ART coverage in high burden areas.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file."} +{"text": "Taxodium distichum) trees [1]. We include putative names when possible, guided by the nearest match in the NCBI databank. This data table shows only one representative of each OTU group and it's nearest match in the NCBI databank, along with information about coverage and percent match of the reads. In total there were 144 fungal cultures sequenced, and all sequences were deposited in the NCBI database under accession numbers KY765150\u2013KY765293 [1].We show the distribution of fungal operational taxonomic units (OTUs) cultured from leaves and galls of baldcypress ( Specifications TableValue of the data\u2022These data are the first observations of fungi living in galls of baldcypress midges.\u2022The sequences and putative identifications can be used for future comparisons in other regions.\u2022Some of the fungi may be pathogenic on the midges, which may be pests.12We amplified the nuclear ribosomal internal transcribed spacer (nrITS) and partial large subunit using primers s ITS1F (5\u2032-CTTGGTCAT TTAGAGGAAGTAA) or ITS5 (5\u2032-GGAAGTAAAAGTCGTAACAAGG) for the forward reaction and LR3 (5\u2032-GGTCCGTGTTTCAAGAC) or ITS4 (5\u2032-TCCTCCGCTTATT GATATGC) for the reverse reaction. Our cycling protocol for amplification followed After visualization confirmation with SYBR on 1% agarose gels, we sent the PCR products to Beckman Coulter Genomics for Sanger sequencing. We sent 164 PCR products from individual cultures of gall or foliar fungi. Due to low quality in the sequences, 20 samples were removed before analyses, leaving 144 sequences. The sequences were assembled and edited in Sequencher v5.0 with support from Mesquite. Operational taxonomic units (OTUs) were formed from sequences assembled based on 97% similarity . These O"} +{"text": "Bacillus subtilis is a Gram-positive bacterium that serves as an important experimental system. B.\u00a0subtilis NCIB 3610 is an undomesticated strain that exhibits phenotypes lost from the more common domesticated laboratory strains. Here, we announce the complete genome sequence of DK1042, a genetically competent derivative of NCIB 3610. Bacillus subtilis strain NCIB 3610 (abbreviated as 3610) is a derivative of Marburg with genomic similarity to B.\u00a0subtilis 168 strain, facilitating genetic study of the complex phenotypes associated with undomesticated strains and consisted of 84 contigs (Genomic DNA from DK1042 (NCIB 3610 omIQ12L) . Sequencrom AMOS . The ori contigs . Thus, tB.\u00a0subtilis subsp. subtilis strain NCIB 3610 comIQ12L was deposited in the DDBJ/EMBL/GenBank database with accession no. CP020102 for the chromosome and CP020103 for pBS32 (Gene annotation was performed using the Prokaryotic Genomes Annotation Pipeline (PGAP) through NCBI. The complete genome sequence of"} +{"text": "TOP2A (topoisomerase (DNA) II alpha), CDK1 (cyclin-dependent kinase 1), CCNB1 (cyclin B1), PCNA (proliferating cell nuclear antigen), MAD2L1 (mitotic arrest deficient 2 like 1), BUB1 (budding uninhibited by benzimidazoles 1 homolog), CCNB2 (cyclin B2), AURKA (aurora kinase A), CCNA2 (cyclin A2), CDC6 (cell division cycle 6 homolog), were identified from protein-protein interaction network. Furthermore, Module analysis revealed that the ten hub genes except TOP2A were belonged to module 1, indicating the upregulation of these hub genes associated molecular pathways in nasopharyngeal carcinoma might activate nasopharyngeal carcinoma pathogenesis. In conclusion, this study indicated that the identified differentially expressed genes and hub genes enrich our understanding of the molecular mechanisms of nasopharyngeal carcinoma, which could eventually translate into additional biomarkers to facilitate the early diagnosis and therapeutic approaches.Nasopharyngeal carcinoma is a metastatic malignant tumor originating from nasopharyngeal epithelium. Lacking or nonspecific symptoms of patients with early stage nasopharyngeal carcinoma have significantly reduced the accuracy of diagnosing and predicting nasopharyngeal carcinoma development. This study aimed to identify gene signatures of nasopharyngeal carcinoma and uncover potential mechanisms. Two gene expression profiles (GSE12452 and GSE13597) containing 56 nasopharyngeal carcinoma samples and 13 normal control samples were analyzed to identify the differentially expressed genes. In total, 179 up-regulated genes and 238 down-regulated genes were identified. Functional and pathway enrichment analysis showed that up-regulated genes were significantly involved in cell cycle, oocyte meiosis, DNA replication and p53 signaling pathway, while down-regulated genes were enriched in Huntington's disease,metabolic pathways. Subsequently, the top 10 hub genes, Nasopharyngeal carcinoma (NPC) is the most common squamous cell carcinoma arising from nasopharynx. Worldwide, nasopharyngeal carcinoma is predominant in east and southeast parts of Asia, south-central Asia, and north and east Africa . In addiIn recent years, the high-throughput platforms for analysis of gene expression, such as microarray technologies, has been broadly used to obtain general genetic alteration during tumorigenesis , 7. Manyhttp://www.ncbi.nlm.nih.gov/geo/geo2r/) to screen differentially expressed genes (DGEs) between nasopharyngeal carcinoma and normal samples. A total number of 56 nasopharyngeal carcinoma samples and 13 normal control samples were analyzed. The top 250 DEGs were respectively screened out in GSE12452 and GSE13597 datasets based on GEO2R. Of which, 417 DEGs lists were finally identified using FunRich_V3 software [https://software.broadinstitute.org/morpheus/) were depicted in Figure The raw data file of GSE12452 and GSE13597 were uploaded to GEO2R II alpha), CDK1 (Cyclin-dependent kinase 1), CCNB1 (cyclin B1), PCNA (proliferating cell nuclear antigen), MAD2L1 (mitotic arrest deficient 2 like 1), BUB1 (budding uninhibited by benzimidazoles 1 homolog), CCNB2 (Cyclin B2), AURKA (Aurora kinase A), CCNA2 (Cyclin A2), CDC6 (cell division cycle 6 homolog). Among these genes, TOP2A and CDK1 showed higher node degrees, which were respectively 66 and 61. In addition, the top three significant modules were obtained from PPI network of DEGs using MCODE analyzing network provides valuable information for understanding cellular functions and biological processes. The top 10 hub nodes with higher degrees were screened out from the PPI network of DEGs consisted of 374 nodes and 1218 edges based on the information in the STRING database. These hub genes included g Figure . FurtherDespite advances in radiation technology, distant metastasis was still the major pattern of treatment failure of NPC. More effective treatment modalities to reduce the rate of distant metastasis attract tremendous attentions . TherefoTOP2A (topoisomerase (DNA) II alpha), a nuclear enzyme which is involved in cell division and cell cycle, was identified as one of the hub genes exhibiting the highest degree of connectivity in current study. Kaplan et al. [TOP2A represented a direct molecular target of anthracyclin-based chemotherapy and topoisomerase inhibitor, such as etoposide. It has also reported that TOP2A was the well-known good prognostic marker in breast cancers, which was associated with a favorable response to anthracyclin-based therapy [TOP2A significantly correlated with more advanced American Joint of Cancer Committee (AJCC) stages and independently predicted worse disease-specific survival (DSS) and distant metastasis-free survival (DMFS) in nasopharyngeal carcinoma. In addition, TOP2A overexpression was also reported in other cancer types, such as prostate cancer [TOP2A in this study was identified as one of the top-ranking upregulated candidates among the differentially expressed genes in NPC tissues, which is demonstrated by previous research. The second hub gene cyclin-dependent kinase 1 (CDK1) that controlling cell cycle events including DNA replication and segregation, transcriptional programs and cell morphogenesis have been identified. It was suggested that CDK1 played a significant role in the control of the eukaryotic cell cycle by restricting the centrosome cycle as well as mitotic onset [CDK1/cyclin B1 pathway might serve as a promising therapeutic target for NPC treatment [CDK1 in tumorgenesis was postulated in various types of cancer, including laryngeal cancer [CCNB1) could activate CDK1, which controlled key early mitotic events including growth inhibition and S-G2/M phase arrest [CCNB1 was found in many different diseases, including colorectal cancer, breast, pancreatic cancer, and meningioma [CCNB1 was also found in our study, which suggested that the upregulation of CDK1/cyclin B1 pathway might activate cell cycle progression in NPC.Based on the PPI network, 10 hub genes that can highlight the further insight of NPC development at molecular level for therapeutic studies were obtained. n et al. demonstr therapy \u201316. More therapy reportede cancer , endomete cancer , colorece cancer , and lune cancer , and so ic onset . In addireatment . Moreovel cancer and ovarl cancer . Increase arrest . Tulalame arrest reportede arrest . Overexpningioma \u201332. OverCCNA2) and cyclin B2 (CCNB2) that related to cell cycle at the G2/M (mitosis) transition and affected chromosomal stability were identified. It was reported that Cyclin A2 (CCNA2) was significantly overexpressed in various cancer types including ER+ breast cancer [CDKs including CCNA2 and Rb/E2F signaling pathway. And Huang et al. [CCNB2 tuned G2/M transitions time though regulating a Golgi checkpoint. However, cyclin B2 overexpression was a poor prognostic biomarker in non-small cell lung cancer [The other two members of cyclin family cyclin A2 genes has an essential role in the control of eukaryotic DNA replication by increasing the polymerase's processibility during elongation of the leasing strand. Recent evidence has demonstrated that PCNA was a key factor in cell cycle regulation and DNA replication via interacting with cell cycle-regulated proteins [PCNA were found to be prognostic and diagnostic implications of salivary gland cancers [PCNA was not a useful marker for advanced NPC, which did not predict the results of treatment in patients. For aurora kinase A (AURKA), one of mitotic serine/threonine kinases contributes to the regulation of cell cycle progression, which associates with centrosome and the spindle microtubules during mitosis and plays a critical role in various mitotic events including the establishment of mitotic spindle, centrosome separation as well as maturation, formation and function of the bipolar spindle, and cytokinesis. Genetic amplification and mRNA and protein overexpression of AURKA are common in many different tumors, which have allowed aurora kinase A to be a potential target for development of cancer therapeutics in early-phase clinical trials [Expression of the proliferating cell nuclear antigen is involved in the initiation of DNA replication and participates in checkpoint controls that ensure DNA replication is completed before mitosis is initiated. Huang et al. [CDC6 also activated ribosomal DNA transcription initiation during cell cyle at G1/S (start) transition. High expression of CDC6 in epithelial ovarian cancer (EOC) cells was indicated as a new potential therapeutic target for EOC patients [CDC6 as a key regulatory target for tumorgenesis was postulated in various types of cancer including prostate cancer [CDC6 in NPC remain unclear. In this study, serine/threonine-protein kinase BUB1(budding uninhibited by benzimidazoles 1 homolog) that performs two crucial functions during mitosis was identified as one of the hub genes. Moreover, mitotic arrest deficient 2 like 1 (MAD2L1) that is required for the execution of the mitotic checkpoint which monitors the process of kinetochore-spindle attachment and inhibits of the anaphase promoting complex was also identified as hub gene. Taken together, these data suggested that the hub genes overexpressed and highly connected may be involved in the regulation of mitosis, and cell cycle process of NPC via the checkpoint mechanism [Cell division cycle 6 homolog . GSE12452 was based on platform GPL570 (Affymetrix Human Genome U133 Plus 2.0 Array). Total RNA extracted from snap frozen laser-captured epithelium from 31 patients with Epstein-Barr virus positive undifferentiated nasopharyngeal carcinomas and 10 patients with no evidence of malignancy. mRNA expression levels were measured for essentially all human genes and all latent Epstein-Barr virus (EBV) genes in nasopharyngeal carcinoma tissue samples and normal nasopharyngeal tissues. Data were analyzed for differential gene expression between tumor and normal tissue and for correlations with levels of viral gene expression. GSE13597 which was based on platform GPL96 (Affymetrix Human Genome U133A Array) consisted of 25 EBV-related nasopharyngeal carcinoma samples and three normal control samples. Total RNA extracted from snap frozen nasopharyngeal biopsies from 25 patients with Epstein-Barr virus positive undifferentiated NPC and 3 patients with no evidence of malignancy.Two gene expression profiles (GSE12452 and GSE13597) were downloaded from GEO datasets is a useful method to compare two or more groups of samples in order to identify genes that are differentially expressed across experimental conditions. We applied the adjusted P values (adj. P) to correct for the occurrence of false positive results using Benjamini and Hochberg false discovery rate method by default. We used log transformation to identify DEGs with a change twofold and defined a P value cut-off of <0.01 to be statistically significant.GEO2R (http://david.abcc.ncifcrf.gov/) is a bioinformatics enrichment web tool for researchers to gain comprehensive high-throughput gene functional annotation analysis.[The Database for Annotation, Visualization and Integrated Discovery is an online program aimed to provide functional protein association networks. The newest STRING version 10.0 covers millions of proteins from more than 2000 organisms [The Search Tool for the Retrieval of Interacting Genes (STRING) database (rganisms . In orderganisms . The APP"} +{"text": "CoTe nanorods exhibit weak ferromagnetism while CoTe2 nanorods present paramagnetic behavior. Different magnetic behaviors occur in the other CoTe2 nanostructures due to Na+ entrance into CoTe2 crystals. A first-principles study on Na-doped CoTe2 confirms the magnetic characteristics.CoTe and CoTe This is consistent with previous reports , which is accordance with the other CoTe nanostructures crystallographic orientation, which is similar to CoTe2 nanotubes ; Fundamental Research Funds of North Minzu University [grant number 2014XBZ02]."} +{"text": "Pre-exposure prophylaxis (PrEP) with emtricitabine and tenofovir disoproxil fumarate (FTC/TDF) is a novel HIV prevention strategy. Suboptimal PrEP adherence and HIV infection creates an opportunity for continued antiretroviral drug activity during undiagnosed infection. We previously showed that macaques infected with SHIV during PrEP with FTC/TDF display reduced acute plasma viremias and limited virus diversity. We investigated the effect of PrEP on acute SHIV DNA dynamics and on the size of the persistent virus reservoir in lymphoid tissues.Cell-associated SHIV DNA levels in PBMCs were measured in 8 macaques infected during PrEP with FTC/TDF or single-agent TAF and was compared to those seen in untreated infections (n = 10). PrEP breakthrough infections continued treatment with 1\u20132 weekly drug doses to model suboptimal drug exposure during undiagnosed HIV infection in humans. SHIV DNA was also measured in lymphoid tissues collected from FTC/TDF PrEP breakthroughs after 1 year of infection.Compared to untreated controls, PrEP infections had reduced plasma RNA viremias both at peak and throughout weeks 1\u201312 (p<0.005). SHIV DNA levels were also reduced at peak and during the first 12 weeks of infection (p<0.043) but not throughout weeks 12\u201320. At 1 year, SHIV DNA reservoirs in lymphoid tissues were similar in size among macaques that received PrEP or placebo.Antiviral drug activity due to PrEP limits acute SHIV replication but has only a transient effect on cell-associated SHIV DNA levels. Our model suggests that suboptimal drug exposure in persons that are taking PrEP and become infected with HIV may not be sufficient to reduce the pool of HIV-infected cells, and that treatment intensification may be needed to sustain potential virological benefits from the PrEP regimen. Daily pre-exposure prophylaxis (PrEP) with tenofovir disoproxil fumarate (TDF) in combination with emtricitabine (FTC) is an effective strategy to prevent HIV infection. In clinical trials, the level of HIV protection by PrEP was strongly correlated with drug adherence . In the Suboptimal PrEP adherence and HIV infection creates an opportunity for continued antiretroviral (ARV) drug activity during undiagnosed infection. Transient drug activity between HIV infection and diagnosis may potentially reduce virus replication. In macaques, infection with a chimeric simian HIV (SHIV) during concurrent PrEP failure with FTC/TDF resulted in blunted acute plasma viremias documented sequentially by weekly sampling following first viral RNA detection , 7. In aEmerging evidence shows that early ARV treatment has greater impact in limiting HIV or SIV reservoirs than later treatment but the long-term effect on virus control or total reservoir size is not fully clear especially when ARV treatment is stopped \u201314. HereAll animal procedures were performed according to NIH guidelines and approved by the Institutional Animal Care and Use Committee (IACUC) of the Centers for Disease Control and Prevention (CDC). Macaques were housed at the CDC under the care of CDC veterinarians in accordance with the Guide for the Care and Use of Laboratory Animals 8th Ed. All procedures were performed under anesthesia , and all efforts were made to minimize suffering, improve housing conditions, and to provide enrichment opportunities. For housing, macaques were maintained in cages that met or exceeded the minimum size requirements as stipulated in the Guide for the Care and Use of Laboratory Animals [cage dimensions (inches): 30 (height) x 30 (width) x 30 (length)]. Steps were taken to reduce animal suffering, which included providing enrichment opportunities [e.g. cage features like swings/perches and objects for the macaques to manipulate], an assortment of food selections like fruits, vegetables or seeds, suitable feeding methods (foraging and task-oriented), and humane interactions with caregivers and research staff. All animals have access to clean, fresh water at all times. Commercial diets are specifically formulated to meet vitamin C requirements. Prior to the initiation of virus inoculations, compatible macaques are pair-housed. Once inoculations are initiated, the macaques are separated into single housing (while permitting eye-contact) with a cage divider to prevent the possibility of SHIV transmission between the macaques. If one macaque remained uninfected during the course of a study, the animal remained separated from other infected macaques, and was not be pair-housed with an infected macaque during the follow-up period. Euthanasia of SHIV infected macaques 162P3 once a week for up to 14 weeks until infection was confirmed by molecular and serologic testing [A first cohort of macaques comprised animals enrolled in SHIV transmission studies as PrEP-treated (n = 8) or controls (n = 10) , 7, 15. A second cohort of 17 animals was used to investigate the long-term effect of early PrEP treatment. We quantified cell-associated SHIV DNA in lymphoid tissues collected at necropsy approximately 1 year after confirmed infection. Of these animals, 9 were infected during intermittent PrEP with FTC/TDF and 8 were untreated controls. Because of limited specimen availability, only a few animals from the first cohort were included in this second analysis. We thus included additional animals that also received FTC/TDF (1 or 2 doses every week) during the first months (median = 3.7) of infection , 16 Tab. Some ofRhesus macaque PBMCs were obtained from blood collected in BD Vacutainer\u00ae CPT\u2122 Cell Preparation Tubes (CPT) containing sodium heparin, resuspended in 200 ul of PBS, and stored at -80\u00b0C until use. Genomic DNA was extracted from 200 ul aliquots using the QIAamp DNA Mini and Blood Mini kit (Qiagen) and resuspended in 100 ul of elution buffer. Total SHIV DNA was quantified using a novel DSP assay coupled with co-amplification of RNase P gene as internal PBMC control . Amplifi162P3 DNA and cellular RNase P were quantified using standard curves prepared by serially diluting known copies of plasmid pVP1 and known amounts of macaque PBMCs, respectively. Plasmid pVP1 contains the 5\u2032 portion of SIVmac239 from the 5\u2032 long terminal repeat to a Sph I restriction site at nucleotide 6707, and was kindly provided by Dr. Cecilia Cheng-Mayer (Aaron Diamond AIDS Research Center). The concentration of pVP1 was determined using a Nano Drop ND-1000 Spectrophotometer. Control PBMCs were prepared from uninfected rhesus macaques and counted using the Guava Personal Cell Analysis (PCA) system. SHIV162P3 RNA was quantified using a modification of an existing two-step RT-PCR assay [5) from an HIV-1CM240 virus stock (NIH Reference Reagent Program) was added to each sample prior to RNA extraction.SHIVCR assay that inc162P3 (gag) and HIV-1CM240 (env) sequences was done using primers specific for SIVmac239 and HIV-1CM240, respectively. SIV gag sequences were amplified using primers SIVp15f1 5\u2019 GCC AAC AGG CTC AGA AAA TTT AA 3\u2019 and SIVp15r (5\u2019 TCC TCA GTG TGT TTC ACT TTC TCT TC 3\u2019) with internal probe P12P HEX (5\u2019HEX AGC CTT TAT AAT(BHQ1) ACT GTC TGC GTC ATC TGG TGC BHQ1 3\u2019). Control HIV1CM240 was amplified using primers envE2f (5\u2019 -GGA CAG GGC CAT GTA AAA ATG T -3\u2019) and EnvE2r (5\u2019 -TCT TCT GCT AGA CTG CCA TTT AAC AG -3\u2019) with internal probe envEP (5\u2019 FAM CAC ACA TGG AAT(BHQ1) TAA GCC AGT GRT ATC MAC TCA BHQ1 3\u2019). RT-PCR was done using the Superscript III one-step qRT-PCR kit (Invitrogen). The final concentrations of SIVmac239 primers and probe in the reaction were 200 nM. Likewise, final concentrations of HIV-1CM240 primers and probe were 25nM and 100nM, respectively. Cycling conditions were 10 minutes at 25\u00b0C, 60 minutes at 48\u00b0C, and 45 cycles of 95\u00b0C for 15 seconds and 60\u00b0C for 1 minute. The sensitivity of the RT-PCR assay is 50 RNA copies/ml [Reverse transcription and PCR amplification of SHIVopies/ml , 20.Peak RNA and DNA levels, area under the curve (AUC) values for RNA and DNA during 5 or 20 weeks, and tissue DNA levels in PrEP breakthrough and placebo controls were compared using a two-tailed Wilcoxon rank-sum test. All analyses were done in GraphPad software (version 5.04). The relationship between plasma RNA and cell-associated DNA levels was determined using the Pearson correlation coefficient.162P3 without any PrEP intervention log10 DNA copies/106 PBMCs and reached a set-point generally within 5 weeks of infection. During these 5 weeks, DNA and RNA levels were highly correlated (162p3 is characterized by a rapid control of plasma viremia within 12 weeks of infection and the establishment of a DNA reservoir of about 102 to 104 SHIV DNA copies per 106 PBMCs.We first defined acute plasma RNA and cell-associated SHIV DNA dynamics in rhesus macaques (n = 10) infected rectally with SHIVrvention . The medrrelated ; PearsonTo investigate if PrEP can reduce the levels of cell-associated SHIV DNA in PBMCs, we compared acute SHIV DNA dynamics between 8 PrEP breakthrough infections and the 10 untreated controls. These macaques were infected after a median of 5 (range = 2\u201314) rectal challenges which was not different from the median of 3 (range = 1\u201310) rectal challenges required to infect the 10 control animals (p = 0.099). Six of the 8 breakthrough infections occurred during intermittent PrEP with FTC/TDF (2 FTC/TDF doses per week) and two infections happened during intermittent PrEP with TAF . The 8 PrEP breakthrough infections continued receiving intermittent antiretroviral treatment after confirmed infection for an average of 4.7 months . Treatme162p3 infections, RNA and DNA levels were highly correlated during the first 5 weeks of infection with the only exception of macaques 35838 and 33053; Pearson correlation coefficients were very strong in 5 macaques and strong in 1 animal (FCW).162p3 as well as the more chronic phase when viral RNA set-points are reached were significantly lower than in untreated controls (p = 0.043). DNA levels during the first 12 weeks of infection were also lower in the animals that received PrEP . (0-12wk) were also significantly lower in PrEP breakthrough infections compared to untreated infections . . However= 0.274) . These ry, 162P3 .0-12wk or AUC12-20wk values and cumulative number of virus challenges (Pearson correlation coefficients of -0.128 (p = 0.61) and -0.384 (p = 0.115), respectively).We also evaluated if the overall SHIV DNA levels were related to the number of challenges needed to infect a macaque. No significant correlation was found between AUC0-12wk values in these additional PrEP breakthrough infections were also reduced compared to untreated controls (p = 0.015 for each comparison).We next investigated the effect of PrEP on the size of the virus reservoir in lymphoid tissues in a late-stage of infection. We quantified cell-associated SHIV DNA in mesenteric, axillary, and inguinal lymph nodes collected from 9 PrEP breakthrough and 8 placebo infections at necropsy. The specific PrEP interventions and dosing regimens, the length of treatment and time between SHIV infection and necropsy, and the levels of SHIV DNA for each lymph node location are shown in 10 SHIV DNA copies/106 cells; range = 1.6\u20133.7) or placebo (p = 0.34) . Levels of SHIV DNA in inguinal and mesenteric lymph nodes were comparable among the 2 groups of macaques . When inmacaques .We investigated in macaques if transient drug activity during PrEP breakthrough infection has the potential to alter the pool of SHIV-infected cells. Our analysis included macaques that received non-daily PrEP regimens with either FTC/TDF combination or TAF, and that continued receiving treatment with 1\u20132 weekly pills during several months. This scenario can easily mimic newly acquired HIV-1 in persons receiving PrEP due to inconsistent adherence and who continue on the PrEP regimen until diagnosis. We found that PrEP significantly reduced peak SHIV DNA levels in PBMCs, although the overall effect on the cell-associated DNA reservoir was only transient and was lost during the first year of infection. Our findings differ from studies of acute treatment with fully suppressive regimens which have generally observed substantial reductions in cell-associated HIV or SIV DNA in PBMCs and lymphoid tissues \u201314. We s95 in macaques [6 cells) TFV-DP concentrations in PBMCs for up to 7 days [Our analysis was retrospective and included a heterogeneous group of macaques that received FTC/TDF or TAF at different times relative to virus challenges. However, most of the FTC/TDF failures received 2 weekly drug doses which is sufficient for accumulating TFV-DP in PBMCs at levels above the prophylactic ECmacaques . The doso 7 days . Thus, aIn contrast to persons who fail PrEP because of low medication adherence and drug levels, our study included animals who represented rare PrEP failures that had drug concentrations in PBMCs that were indistinguishable from protected animals , 7. For 162p3, we found no correlation between cumulative exposures to SHIV and peak plasma RNA levels or AUC values [Although not statistically significant, we found that the number of SHIV challenges required to infect untreated macaques was smaller than those required to infect the animals that failed PrEP, which raises questions about the effect of previous SHIV exposures on systemic control of the virus post-infection. In a large retrospective analysis of 40 rhesus macaques exposed repeatedly to SHIVC values . In our 162p3 is easily transmissible at low doses and is thus well-suited for transmissibility studies, infections with this isolate generally result in low set-point viremias compared to more pathogenic SIV isolates. Also, the infection course is generally non-pathogenic [The present study has several limitations. First, we did not address potential effects of PrEP on the fraction of DNA genomes that are replication-competent. Reduced virus replication and evolution due to PrEP might also result in more genetically homogeneous virus reservoirs or favor preferential seeding of sequences with replication advantages in the presence of FTC or TFV . Second,thogenic , 26 whicIn summary, we show that transient antiviral drug activity due to PrEP limits acute SHIV replication but only has a transient effect on the size of the DNA reservoirs. These observations suggest that suboptimal drug exposure in persons that are taking PrEP and become infected with HIV may not be sufficient to durably reduce the pool of HIV-1-infected cells and, therefore, not likely to predict good virus control post PrEP interruption.S1 FigA) Serial dilutions of SIV vP1 DNA plasmid were prepared in PBS and tested 10 times in the same run. The assay consistently detected 10 copies of vp1. (B) Variability in Ct values observed in 16 separate runs at different inputs of SIVmac239 vp1 plasmid and rhesus PBMCs. In all instances the % coefficient of variation (CV) was <4%.(TIF)Click here for additional data file."} +{"text": "This article contains data related to the research article entitled \u201cDistribution and potential ecological risk of 50 phenolic compounds in three rivers in Tianjin, China\u201d [1]. This data article reports the detailed information for the contaminant level of phenolic compounds in three rivers in Tianjin, China. The data collects from seven sample sites in Beitang drainage river, sixteen sample sites in Dagu drainage river, and fourteen sample sites in Yongdingxin river. The ranges, standard deviations, average values, median values of the concentrations of identified phenolic compounds in three rivers and the standard deviations, average values, the maximum values of risk quotients of identified phenolic compounds in three rivers are listed in this paper. Specifications tableValue of the data\u2022The data provide more details on distribution of phenolic compounds in rivers in Tianjin.\u2022The data present here will be valuable for ecological risk assessment of phenolic compounds in water environment.\u2022The data can be used for the water environmental managers for proper operation.1The detailed information of 50 phenolic compounds are displayed in our previously papers 22.1Thirty-seven sample sites were set in three rivers. b1\u2013b7 were situated in Beitang drainage river (BDR), d1\u2013d16 were located in Dagu drainage river (DDR) and y1\u2013y14 were located in Yongdingxin river (YDXR). Thirty-seven surface water samples, thirty-seven SPM samples and thirty-six sediment samples (excluding b6) were collected in the wet season. Twenty-nine surface water samples, suspended particulate matter (SPM) samples and sediment samples were collected in dry season .2.2The pesticide-residue grade n-hexane and dichloromethane were purchased from Mallinchrodt Baker, Inc. (USA). Derivatization reagent N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) with 1% trimethylchlorosilane (TMCS) was purchased from Supelco Co. (USA). Glass fiber filter membranes were purchased from Millipore Co. The C18 cartridges and Oasis HLB cartridges were purchased from Supelco Co. and Waters (USA), respectively.2.3The chemicals and materials used to treat and analyze samples, the detailed methods for preparing water samples and analytical procedures have been published elsewhere"} +{"text": "Esr2 exon 3 deletion (\u22063) and another DNA binding domain (DBD) mutant with exon 4 deletion (\u22064) were compared to that of wildtype (WT) rats. The raw data were analyzed using CLC genomics workbench. High quality RNA-sequencing reads were aligned to the Rattus norvegicus genome. Differentially expressed genes in \u22063 or \u22064 Esr2-mutant GCs were identified based on the following criteria: FDR p-Value \u22640.05 and an absolute fold change of 2. Fewer differentially expressed genes were identified in \u22063 compared to the \u22064 mutant group. As both mutant groups demonstrated a common phenotype of ovulation failure, differentially expressed genes common to both in \u22063 and \u22064 mutant rats were emphasized and further analyzed in the companion article \u201cESR2 regulates granulosa cell genes essential for follicle maturation and ovulation\u201d RNA seq analyses were performed in granulosa cells (GCs) collected from gonadotropin treated ESR2 mutant rats. Data obtained from a null mutant with Specifications TableValue of the data\u2022This study provides the transcriptomic analyses of granulosa cells of ESR2 mutant rats (\u22063 and \u22064).\u2022Esr2-mutant female rats from both groups were infertile due to failure of ovulation. Differentially expressed genes in Esr2 mutant GCs may represent the ESR2 target genes downstream of gonadotropin induced ovulation.1\u2022\u2022\u2022\u2022\u202222.1Esr2-mutant rat models were generated by targeting exon 3 (\u22063) or exon 4 (\u22064) in the Esr2 gene as described previously All procedures were performed in accordance with the protocols approved by the University of Kansas Medical Center Animal Care and Use Committee. Holtzman Sprague-Dawley (HSD) 2.2Esr2-mutant (\u22063 and \u22064) and age-matched wildtype (WT) female rats were used for the gonadotropin induced follicular development. Synchronized follicular growth was initiated by intraperitoneal injection of 30 IU Pregnant Mare\u05f3s Serum Gonadotropin . 48\u202fh after the PMSG injection, 30 IU of Human Chorionic Gonadotropin was injected intraperitoneally.Four-week-old 2.3Animals were sacrificed 10\u202fh after exogenous gonadotropin administration. GCs were collected from the gonadotropin treated WT, \u22063 and \u22064 ovaries, and total RNA was extracted by using TRI Reagent (Millipore-Sigma) following the manufactures instruction. RNA quantification was performed by using nanodrop (Thermo Scientific) and approximately 500\u202fng of total RNA was used for the RNA-seq library preparation. Libraries were prepared by using TruSeq standard mRNA kit (Illumina) following the manufacturer\u05f3s instruction. The cDNA libraries were sequenced at the Molecular Biology Core Laboratory of Mayo Clinic .2.4Rattus norvegicus genome (downloaded from NCBI database) using default parameters: (a) maximum number of allowable mismatches was 2 (b) minimum length and similarity fraction was set at 0.8; and (c) minimum number of hits per read was 10. A total of 32,623 genes were detected in each group of GCs. Expression values were measured in RPKM (Reads per kilobase of exon model per million mapped reads) RNA-Seq data were analyzed by using the CLC Genomics Workbench (Qiagen Bioinformatics) to identify the differentially expressed genes. All clean reads were obtained by removing low quality reads by trimming, and the high-quality reads were aligned to the 3For RNA Seq, each study group contained three library samples. Each library sample was made by pooling two RNA samples from two individual rats from the same genotype. In CLC Genomics Workbench, the \u2018Differential Expression for RNA-Seq tool\u2019 performs some multi-factorial statistics on a set of Expression Tracks based on a negative binomial Generalized Linear Model (GLM). The final GLM fit and dispersion estimate calculate the total likelihood of the model given the data, and the uncertainty on each fitted coefficient. Two statistical tests- Wald test and Likelihood Ratio test, each make use of one of these values. The Likelihood Ratio test is used in the Across groups (ANOVA-like) comparison."} +{"text": "Escherichia coli (EPEC/EHEC) manipulate a plethora of host cell processes to establish infection of the gut mucosa. This manipulation is achieved via the injection of bacterial effector proteins into host cells using a Type III secretion system. We have previously reported that the conserved EHEC and EPEC effector EspG disrupts recycling endosome function, reducing cell surface levels of host receptors through accumulation of recycling cargo within the host cell. Here we report that EspG interacts specifically with the small GTPases ARF6 and Rab35 during infection. These interactions target EspG to endosomes and prevent Rab35-mediated recycling of cargo to the host cell surface. Furthermore, we show that EspG has no effect on Rab35-mediated uncoating of newly formed endosomes, and instead leads to the formation of enlarged EspG/TfR/Rab11 positive, EEA1/Clathrin negative stalled recycling structures. Thus, this paper provides a molecular framework to explain how EspG disrupts recycling whilst also reporting the first known simultaneous targeting of ARF6 and Rab35 by a bacterial pathogen.Enteropathogenic and enterohaemorrhagic \u2022EHEC delivers effector proteins into host cells to establish infection in the gut\u2022The effector EspG interacts with GTP-ARF6 confining EspG to recycling endosomes\u2022During infection EspG interacts preferentially with Rab35, not Rab1\u2022Spatial restriction of bacterial effectors during infection determines their function Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC) use a type III secretion system (T3SS) to deliver an array of bacterial effector proteins into host cells during infection, facilitating colonization of the gut epithelia The attaching and effacing (A/E) pathogens enterohaemorrhagic Recently we demonstrated that EHEC depletes a number of cell surface receptors from the Plasma Membrane (PM) during infection, in a manner dependent on the T3SS effector EspG Since its discovery in 2001 Our investigation of the small GTPase interacting partners of EspG during infection reveals that EspG modulates an ARF6:Rab35 signaling axis to disrupt recycling endosome function, resulting in the accumulation of recycling cargo within the host cytosol. Our results highlight the importance of spatial restriction of bacterial effector proteins during infection, whilst simultaneously providing a molecular mechanism to support previously published EspG phenotypes.In vitro data indicates EspG can interact with at least 3 of the 5 ARF proteins espG\u00a0+\u00a0pEspG:4xHA or constitutively active, GTP-bound (Q67L) ARF6 mutants ARF-binding has been proposed to spatially restrict EspG within host cells to allowing targeted, local Rab inactivation In vitro EspG was shown to act as a Rab GAP for only 12 of the 30 Rabs tested, including, in our panel, Rab 13, 30, 35, 37 and 38 in vitro, EspG showed the highest GAP activity for this protein Over 60 human Rab isoforms have been described to date in vitro, spatial restriction of EspG to endosomal compartments during infection appears to direct EspG's GAP activity towards Rab35.Co-immunoprecipitation experiments using our Rab GTPase panel show that EspG is selectively and consistently immunoprecipitated by Rab35 . Peripheral endosomes containing TfR and EEA1 showed similar association with CLC in all conditions Legionella pneumophilia Dot/Icm effector AnkX Whilst a number of reports in the literature have identified ARF6 and Rab35 individually as targets during bacterial infection (ARF6 is implicated in host cell invasion by both"} +{"text": "S. cerevisiae. G58D had been previously proposed to have unique effects on these variants, but our studies suggest a common mechanism. All variants, including those reported to be resistant, are inhibited by G58D but at distinct doses. G58D lowers the kinetic stability of the associated amyloid, enhancing its fragmentation by molecular chaperones, promoting Sup35 resolubilization, and leading to amyloid clearance particularly in daughter cells. Reducing the availability or activity of the chaperone Hsp104, even transiently, reverses curing. Thus, the specificity of inhibition is determined by the sensitivity of variants to the mutant dosage rather than mode of action, challenging the view that a unique inhibitor must be developed to combat each variant.Prions adopt alternative, self-replicating protein conformations and thereby determine novel phenotypes that are often irreversible. Nevertheless, dominant-negative prion mutants can revert phenotypes associated with some conformations. These observations suggest that, while intervention is possible, distinct inhibitors must be developed to overcome the conformational plasticity of prions. To understand the basis of this specificity, we determined the impact of the G58D mutant of the Sup35 prion on three of its conformational variants, which form amyloids in Prion proteins adopt alternative conformations and assemble into amyloid fibers, which have been associated with human disease. These fibers are highly stable and self-replicate, leading to their persistence and resulting in a set of progressive and often fatal disorders. Inhibitors have been shown to interfere with some conformations but not others, suggesting that distinct strategies must be developed to target each. However, we show here that a single dominant-negative mutant can inhibit multiple conformations of the same prion protein through the same pathway but at distinct doses. Thus, the basis of this specificity is sensitivity rather than resistance to the mechanism of inhibition, suggesting that common strategies may be used to target a range of prion conformations. Our studies indicate that \u201cresistance\u201d to G58D can be partially overcome at higher dosage of the mutant, revealing differential sensitivity to the inhibition. G58D reduces the kinetic stabilities of the amyloids associated with the variants, which determines their efficiencies of fragmentation by chaperones [PSI+]Strong variant Sc4 strain promoted the accumulation of red pigment on rich medium, indicating reversal of the prion phenotype , where the pinker colonies on rich medium relative to the wildtype [PSI+]Weak strain indicated a mild inhibition by G58D , we determined the frequencies of [psi-] appearance during mitotic division for each strain. [PSI+] propagation was largely stable at the 2:1 (~0% curing) and 1:1 (~1% curing) ratios of wildtype to G58D for both [PSI+]Sc4 and [PSI+]Sc37, where the colony phenotype was only mildly reversed , arises in a dose-dependent manner. Together, these observations indicate that the previously described \u201cresistance\u201d of [PSI+]Sc37 and [PSI+]Weak to curing by G58D expression reflected their higher threshold for sensitivity rather than their absolute recalcitrance to inhibition by this mutant.To assess whether reversal of the s tested . Thus, [PSI+] variants studied here, in addition to the previously studied [PSI+]Strong variant [PSI+] variants may reflect the impact that this mutant has on the kinetic stability of each. While it has been well-established that Sup35 aggregates in the [PSI+]Sc4 conformation are of lower stability than those in the [PSI+]Sc37 conformation, the relative stabilities of the four variants have not been previously reported Strong and [PSI+]Sc4 variants in comparison with a wildtype strain , a potent inhibitor of the fragmentation catalyst Hsp104 [PSI+]Strong strain [PSI+]Sc4 and [PSI+]Sc37 diploids reduced propagon number by factors of ~2 and ~4, respectively (PSI+] phenotype and the loss of [PSI+] that we observed in these strains (PSI+]Weak increased propagon number by a factor of ~2.5 (PSI+]Weak phenotype at this ratio (PSI+]Weak phenotype upon G58D expression to much higher levels [PSI+] variants studied here (in vivo.To determine how the kinetic destabilization of Sup35 aggregates by G58D impacts the number of heritable prion units (propagons) in maintenance to capture different variants , but we found that this instability is strongly suppressed by heterozygous disruption of HSP104 . In both cases, the mutant is incorporated into wildtype aggregates but capable of destabilizing the amyloid state only when present in excess to wildtype protein, and the efficacy of dominant-negative inhibition is greater for less kinetically stable conformational variants [PSI+]Sc37 in a similarly digested pRS306. The SUP35(G58D) ORF was then subcloned into pRS306-PADH as a BamHI-EcoRI fragment isolated from pRS306-SUP35(G58D) to create pRS306-PADHSUP35(G58D) (SB468).All plasmids used in this study are listed in Oligonucleotides used in this study are listed in PSI+]Sc4 (SY2085) and [PSI+]Sc37 (SY2086) haploid wildtype strains were gifts from J. Weissman. Yeast strains expressing ectopic copies of SUP35 or G58D from URA3 (pRS306) or TRP1 (pRS304)-marked plasmids were constructed by transforming yeast strains with plasmids that were linearized with BstBI or Bsu361, respectively, and by selecting for transformants on the appropriate minimal medium. In all cases, expression was confirmed by quantitative immunoblotting for Sup35. Disruptions of SUP35 were generated by transformation of PCR-generated cassettes using pFA6aKanMX4 as a template with the indicated oligonucleotide primers (HSP104 disruptions were generated by transformation with a PvuI-BamHI fragment of pYABL5 (a gift from S. Lindquist) and selection on minimal medium lacking leucine. Disruptions of NAT1 were generated by transformation of PCR-generated cassettes using pFA6a-hphMX4 as a template with the indicated primers , SDS-PAGE, quantitative immunoblotting and SDS-sensitivity experiments were performed as previously described . To analin vivo dilution, colony-based method [The number of propagons per cell was determined using a previously described d method . For propsi-].Daughters were separated by FACS based on bud-scar labeling. Yeast cells were incubated for 1 h at room temperature in 1\u03bcg/ml Alexa-647 wheat germ agglutinin (WGA) in PBS. After washing twice in PBS, cells with the lowest fluorescence intensity (5%) were sorted as newborn daughter cells, and a sample of this fraction was viewed by fluorescence microscopy to confirm bud scar number. This fraction was also moved to rich medium (1/4 YPD) for color development. For Hsp104 inhibition, sorted fractions were first moved to a minimal medium with 3mM GdnHCl for three hours before being transferred to rich medium. In each case only completely red colonies were counted as Sc4 (A), [PSI+]Sc37 (B) or [PSI+]Weak (C) strains described in Lysates isolated from [(TIF)Click here for additional data file.S2 FigPSI+] variant haploid yeast strains were analyzed by SDD-AGE and immunoblotting for Sup35.Lysates from the indicated [(TIF)Click here for additional data file.S3 FigPSI+]Sc4 (A), [PSI+]Sc37 (B) or [PSI+]Weak (C) strains described in Lysates isolated from [(TIF)Click here for additional data file.S4 FigPSI+]Sc37 (blue) and [PSI+]Weak (red) after treatment with GdnHCl. Error bars represent standard deviation. .The rate of propagon recovery was determined for [(TIF)Click here for additional data file.S5 FigDIC and Alexa-647 WGA fluorescence of bud scars of cells both before and after FACS sorting. Scale bars represent 3\u03bcm.(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.S1 Text(PDF)Click here for additional data file."} +{"text": "Streptococcus agalactiae strains isolated from Camelus dromedarius in Kenya and Somalia. These data are an extension to the group B Streptococcus (GBS) pangenome and might provide more insight into the underlying mechanisms of pathogenicity and antibiotic resistance of camel GBS.We present draft whole-genome sequences of seven Streptococcus agalactiae, also known as Lancefield\u2019s group B Streptococcus (GBS), is an emerging pathogen of serious clinical concern . Previous genomic analysis of these isolates by multilocus sequence typing (MLST) indicated a detached phylogenetic relationship compared to GBS strains of human or bovine origin . DNA was fragmented by ultrasonication using the Covaris S2 instrument . Barcoded libraries were generated with the Ion fragment library kit and Ion Xpress DNA barcode adaptors . Sequencing was performed on an Ion Torrent Personal Genome Machine (PGM) system, with the Ion PGM sequencing 400 kit and the Ion 318 Chip version 2 . After sequencing, single processing and base calling were performed using Torrent Suite 3.6 , and barcode-separated FASTQ files were generated. For sion 4.0 GBS genosion 4.0 . SeqMan sion 4.0 .S.\u00a0agalactiae (S.\u00a0agalactiae.The draft genome sequences of cameloid GBS isolates presented here are a valuable addition to the pangenome of alactiae . These gS.\u00a0agalactiae isolates were deposited in GenBank under BioProject no. PRJNA382326. The accession numbers for each isolate are shown in The annotated draft whole-genome sequences of the seven"} +{"text": "PTEN) loss as a clonal mutation in the case of a 6-year-old boy with a diffuse intrinsic pontine glioma, and incorporated copy number alteration analyses to provide a more detailed understanding of clonal evolution in diffuse intrinsic pontine gliomas. As well, using the PedcBioPortal, we found alterations in PTEN in 16 of 326 (4.9%) cases of pediatric high-grade glioma (3 of 154 (1.9%) brainstem) for which full sequencing data was available. Our data strengthens the association with PTEN loss in diffuse intrinsic pontine gliomas and provides further argument for the inclusion of PTEN in future targeted sequencing panels for pediatric diffuse intrinsic pontine gliomas and for the development and optimization of mTOR/PI3K inhibitors with optimal central nervous system penetration.Improved molecular understanding is needed for rational treatment of diffuse intrinsic pontine gliomas (DIPG). Here, using multi-focal paired tumor and germline exome DNA and RNA sequencing, we uncovered phosphatase and tensin homolog ( Despite improvements in diagnostic accuracy and multimodal treatments, prognosis for patients with DIPG remains dismal with a median survival of around 1 year.4Brainstem tumors comprise 10\u201315% of central nervous system tumors in children.6 DIPG appears to be a molecularly homogenous tumor as compared to adult glioblastoma multiforme (GBM).8 Deep sequencing of multiple sites within tumors has revealed key mutations, which are inferred to be driving events as they are conserved throughout all samples in tumors with multi-focal analysis. Activating point mutations (H3K27M) in the genes encoding the histone H3 histone family member 3A (H3F3A) or histone cluster 1 H3 family member B (HIST1H3B) proteins are now understood to be early oncogenic events in DIPG.9 These mutations are frequently accompanied by driving mutations in genes encoding activin A receptor type 1 , tumor protein p53 (TP53), or phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA).11 Here, using multi-focal paired tumor and germline exome DNA and RNA sequencing, we uncovered phosphatase and tensin homolog (PTEN) loss as a clonal mutation in the case of a 6-year-old boy with DIPG, and incorporated copy number alteration analyses to provide a more detailed understanding of clonal evolution in a DIPG.Improved molecular understanding is needed for rational treatment of DIPG. Pediatric DIPG has been shown to demonstrate intratumoral spatial histologic heterogeneity.12 Approximately 13 months following his initial diagnosis, the patient died from progressive disease. Consent for research autopsy was obtained and research autopsy was performed within hours of patient\u2019s death.A 5-year-old boy presented with repetitive episodes of stumbling and tripping, as well as bilateral ptosis. Magnetic resonance imaging (MRI) of the brain revealed a 4.6\u2009cm, diffusely infiltrating pontine mass. Surgical resection was not recommended due to the location and size of the lesion, and he was treated on a Children\u2019s Oncology Group phase I trial (NCT01922076) with AZD1775 (Wee1 kinase inhibitor) concurrently during radiation. Two months following chemo-radiotherapy, the patient underwent convection-enhanced delivery of 8-H9 antibody (NCT01502917). An interval MRI 1 month later was concerning for progressive disease with recurrence of prior symptoms and development of new motor deficits. He was then treated off-trial for six cycles with the histone deacetylase inhibitor panobinostat, based on pre-clinical data showing its efficacy in H3K27M-mutated DIPG.13The involved brainstem was sectioned into six cross-sections , ACVR1 (p.G328V), and PTEN (p.Q97X), and copy number loss of PTEN in all five samples. Somatic mutations were additionally found across all five sites in glycoprotein VI and adenosine monophosphate deaminase 1 , though these mutations are of uncertain significance. We integrated these and the other mutations alongside copy number variants (CNVs) , and reduced expression of glutamic-oxaloacetic transaminase 1 (GOT1) and proprotein convertase subtilisin/kexin type 9 (PCSK9). While there is no mention of PCSK9 in the context of oncogenesis in the literature, overexpression of CDK18 has been seen in association with breast cancer and reduced expression of GOT1 has recently been demonstrated to be associated with poorer outcomes in glioblastoma.16 Histopathologic analysis of contralateral quadrants from each cross-section revealed pleomorphic cells infiltrating through the pons. No microvascular proliferation or tumor necrosis was observed and the histology was consistent with a diffuse infiltrative glioma with focal anaplastic changes. Tumor cells showed some expression of glial fibrillary acidic protein. An immunostain for Ki-67 showed marked heterogeneity from region to region DIPG tumors, and no cases with mutation or homozygous loss; another identified a PTEN SNV in 1 of 26 (4%) samples.22 Using the PedcBioPortal, which compiled multiple sequencing datasets of human pediatric high-grade glioma and DIPG25 with additional samples deposited by Dr. Chris Jones (EGAS00001001436), we found alterations in PTEN in 16 of 326 (4.9%) cases for which full sequencing is available (3 of 154 (1.9%) brainstem). As well, we found homozygous deletion of PTEN in 7 of 834 (0.8%) in which DNA copy number was available (3 of 242 (1.2%) brainstem). The PI3K/mechanistic target of rapamycin (mTOR)/AKT/PTEN pathway is an attractive target for children with DIPG showing PTEN loss or PI3K activation. The mTOR protein is downstream of PI3K and in vitro data from multiple tumor types with PIK3CA mutations demonstrates sensitivity to mTOR inhibition with the mTOR inhibitor everolimus, which exhibits good CNS penetration.27 Sapanisertib, an agent that inhibits both mTOR complexes 1 and 2, has been shown to be effective against in vitro and murine models of DIPG.28 Recently agents have been explored to dually target PI3K and mTOR, which have shown efficacy in GBM pre-clinically.32PTEN is an established negative regulator of PI3K signaling and tumor suppressor in many human cancers.The prognosis for pediatric DIPG remains bleak despite improved, biopsy-guided multi-modal treatment approaches. It is becoming increasingly evident that a deeper molecular understanding of each individual tumor is needed to guide therapeutic decisions. While our data confirms a relative homogeneity of DIPG in terms of recurrent lesions, our use of copy number analysis and RNA sequencing helps clarify distinct tumor evolution and behavior by region.PTEN loss as an early event in whole exome and transcriptome multi-focal sequencing of a case of DIPG. Our data strengthens its association with DIPG, and provides further argument for the inclusion of PTEN in future targeted sequencing panels for pediatric DIPG and for the development and optimization of mTOR/PI3K inhibitors with optimal CNS penetration.In conclusion, we report Genomic data will be deposited according to established practices of the Michigan Center for Translational Pathology.Supplementary Figure 1. Copy number variant (CNV) and zygosity map for five autopsy sitesSupplementary Figure 2. Histopathology and IHC results for each tumor sample"} +{"text": "Cognitive deficits are considered core features of schizophrenia (SZ) (Green & Harvey 2014). Subtle variations in the perfectly coupled mechanism that maintains the potential of membrane in neurons may have repercussion on neuronal processing. Therefore, genetic variability related to the functioning of excitable cells and linked to pathways essential for neuronal survival and plasticity may underlie the observed differences in cognitive abilities . CACNA1C and KCNH2 genes encode for calcium and potassium voltage-gated channels, ultimately related to neuronal functioning . These two genes have been previously related with SZ . The aim of this study was to evaluate whether the genetic variability of CACNA1C and KCNH2 is associated with: i) the risk for schizophrenia, ii) the cognitive performance of SZ patients and healthy subjects.Our sample consisted of 348 SZ patients and 387 unrelated healthy controls (HC). DNA was extracted from blood/saliva samples using standard procedures and two Single Nucleotide Polymorphisms (SNPs) were genotyped: rs1006737 (G/A) in CACNA1C gene, rs3800779 (G/A) in KCNH2. A subsample (296 SZ/157 HC) underwent neurocognitive assessment, which included: i) premorbid IQ ); ii) memory ) and, iii) executive function ). The association between the SNPs and neurocognitive performance was explored (adjusted by sex and age) separately in patients and in controls groups.In our sample, we did not detect an association of CACNA1C and KCNH2 with the risk for SZ. Patients performed significantly worse than controls in all cognitive measures (p<0.005). SZ patients homozygous for the risk allele (A) of the CACNA1C polymorphism showed lower premorbid IQ (TAP scores) than patients carriers of the C allele (rs1006737: B=-1.39 p=0.027). Within HC, the minor allele (A) of KCNH2 was associated with WMS global score (rs3800779: B=3.01 p=0.010): subjects carrying the AA genotype presented better memory performance.Our findings add evidence on the role of CACNA1C and KCNH2 on modulating cognitive performance in SZ patients and HC . Our results in patients are in line with previous studies that suggest an association of CACNA1C risk allele on different cognitive domains. As regards to KCNH2, our results are opposite in terms of the direction of the effect observed in previous studies, probably as a consequence of the sample size and heterogeneity in methods used to assess memory. The different direction of the genetic effects among patients and controls reflects the complex relationship between genetic factors and cognitive performance variability. It is suggested that genes that enhance cognitive abilities under normal circumstances turn to be pernicious under the modulation effect of a disease . Further research is needed and we expect to extend the present results with neuroimaging genetics approaches.Spanish Ministry of Economy and Competitivity, Instituto de Salud Carlos III (PI15/01420 and PI15/00299), Ayuda cofinanciada por el Fondo Europeo de Desarrollo Regional (FEDER) \u201cUna manera de hacer Europa\u201d."} +{"text": "Gefitinib is an oral EGFR tyrosine kinase inhibitors which may act as a radiosensitizer.EGFR mutation status.This phase II study evaluated the efficacy of gefitinib 250 mg once daily in combination with thoracic radiotherapy followed by consolidation chemotherapy (IV cisplatin and vinorelbine) as first line treatment in a population of unselected stage IIIB NSCLC patients according to n = 50) was not reached. Sixteen patients were included in four centers, 50% had adenocarcinoma and 75% were male. Genomic alterations (7 patients studied) retrieved TP53 mutation in 2 patients and no EGFR mutation. Four weeks after radiotherapy, 3 patients (19%) had a partial response, 6 (38%) had a stable disease, and 7 had a progression (44%). Median overall survival was 11 months and median progression-free survival was 5 months. At the time of the last contact, 5 patients (31%) were still alive. Main toxicities were gastrointestinal (81%), cutaneous (81%), general (56%), and respiratory (50%). There were 12>G3 adverse events in 7 (47%) patients, and there was one toxic-death during the concomitant period due to an interstitial pneumonitis. There were two possible adverse events-related deaths during the chemotherapy period (pulmonary embolism (n = 1) and sudden death after the administration of the 3rd course of chemotherapy (n = 1)).Due to a low accrual rate in this study, the sample size (EGFR mutated NSCLC.The benefit of Gefitinib-RT could not be confirmed due to premature trial discontinuation. Further evaluation is required, especially in patients with EGFR). Mutations in the EGFR tyrosine kinase are observed in approximately in 15% to 62% of patients and are predictors of responsiveness to EGFR tyrosine kinase inhibitors (TKIs) is greater than or equal to 1 L, oxygen diffusion capacity of greater than or equal to 40%); pulmonary dose volume histogram V20Gy inferior or equal to 40%; life expectancy of at least 6 months; Female patients could be included if use of secure contraceptive precautions, or post-menopausal. Exclusion criteria were: prior anticancer treatment (including thoracic radiotherapy or anti-EGFR therapy); known hypersensitivity to gefitinib, cisplatin or vinorelbine, or any of the excipients of these product; interstitial lung disease; malignancies diagnosed within the last 5 years; severe coexisting, psychological, or uncontrolled condition; documented or symptomatic metastases, including positive cytology in the case of pleural effusion; pregnancy or breast feeding; concomitant use of inhibitors of CYP3A4; and weight loss of over 15% in the 3 months before the start of the study. 18F-fluorodeoxyglucose positron emission tomography (18F-FDG PET/CT) and brain magnetic resonance imaging (MRI) were not mandatory for patient inclusion.Patients (\u226518 years) were eligible for inclusion if they had histologically confirmed unresectable non-pretreated stage III NSCLC. Additional inclusion criteria were: at least one measurable lesion according to Response Evaluation Criteria In Solid Tumors (RECIST version 1.0); World Health Organisation (WHO) performance status (PS) of 0 to 2 at inclusion; adequate pulmonary function and vinorelbine (25 mg/m2 once per week for 3 weeks out of 4). We chose cisplatin-vinorelbine doublet for its good efficacy/toxicity profile when delivered sequentially to thoracic radiotherapy .\u201319.EGFR s study) , 21. In mutation . On the months) . Anyway,n = 2), general (n = 2), hematologic (n = 2) and respiratory (n = 2). In our trial, chemotherapy was delivered sequentially because concomitant administration of chemoradiotherapy and gefitinib could have been much more toxic. The triple combination of bioradiotherapy and chemotherapy may lead to unexpected toxicities [The combination of gefitinib and radiotherapy was mild. The most common G3-4 AEs were: gastrointestinal have also been prospectively studied in brain metastatic NSCLC patients with a good overall response rate (86%) and no increased neurotoxicity . A randoEGFR and/or ALK fusion arrangement who receive induction erlotinib or crizotinib before thoracic chemoradiotherapy. Finally, as some of the effects of ionizing radiation are now recognized as contributing to antitumor immunity [Additional anti-EGFR treatments , 31\u201337 aimmunity , 40, tarEGFR mutated NSCLC, our results should be cautiously interpreted. Further investigation is needed to better assess the therapeutic ratio of this combination in trials that take into account modern radiation-delivery techniques and incorporate the biological abnormalities of tumors (EGFR mutated NSCLC).In this phase II trial, gefitinib (250mg once daily) in combination with thoracic radiotherapy administered with conventional fractionation at the dose of 66 Gy followed by chemotherapy in previously untreated stage III NSCLC was feasible but lead to substantial toxicities (one toxic death during the concomitant period and two possible adverse events-related deaths during the chemotherapy period). Due to premature trial discontinuation and the absence of patients with th World Conference on Lung Cancer (WCLC 2015)This work was presented at the 16"} +{"text": "Previous studies measuring cerebral blood flow (CBF) with magnetic resonance sequences like Arterial Spin Labelling (ASL) showed that patients with schizophrenia (SCZ) have increased CBF in basal ganglia and reduced blood flow in cortical areas like the prefrontal cortex. It is still not clear whether these abnormalities are related to antipsychotic treatment or rather they reflect a disease trait independent from medication. Interestingly, administration of single dose of antipsychotics in healthy volunteers produce marked functional effects that are in the same region reported as altered in SCZ. These effects are thought to depend on dopamine D2 receptor (D2R) blockade, although their relationship with antipsychotic pharmacodynamics has not been fully established yet. In fact, the haemodynamic nature of CBF measures makes difficult to interpret drug effects in terms of altered neurotransmission function. Here, we tested whether CBF changes induced by different antipsychotics mirror receptor distribution profiles of D2R. We evaluated the correlation of CBF variation with receptor density as measured with PET and brain mRNA expression extracted from the Allen Human Brain Atlas (ABA).Forty-two healthy male subjects were enrolled in a double blind, randomized, placebo-controlled, crossover study. Participants were randomized in two equal parallel groups to receive a single dose of antipsychotic/placebo in three separate sessions. In Group 1 placebo, olanzapine 7.5mg (OLA) or haloperidol 3mg (HAL) were administered before the MRI scan. In Group 2 participants received placebo, 0.5mg (lowRIS) or 2mg (highRIS) of risperidone. Regional CBF was assessed with pseudo-continuous ASL (pCASL) sequence. For each antipsychotic, a paired T-test was performed in SPM12 with global CBF values as covariate of no interest.A template image of dopamine D2 receptor density was derived from 6 PET scans in healthy volunteers using the high affinity D2/D3 antagonist ligand [18F]-Fallypride. Brain mRNA expression values for DRD2 gene (coding for D2R) were extracted from the ABA dataset by using the MENGA toolbox. CBF contrast images and the [18F]Fallypride BPND template were segmented into 83 ROIs by using the Desikan-Killiany Atlas. The regional changes in CBF against placebo (\u2206CBF) were compared with regional BPND values and gene expression maps using multivariate correlations.For all antipsychotics, CBF changes in each ROI were directly proportional to [18F]Fallypride non displaceable binding potential (BPND) values and DRD2 mRNA expression levels .In the present study, we were able to show that the CBF increase induced by antipsychotic is directly proportional to D2R concentration in the brain, as indexed by PET BPND maps and mRNA expression levels. Interestingly, the association strength between \u2206CBF and brain receptor distribution profiles mirrored differential D2R affinity between the tested drugs. Overall, these results indicate that CBF increases after administration of a single dose of antipsychotics actually reflect known pharmacodynamics profile of these compounds. In addition, these results further reinforce previous evidence suggesting the role of D2R blockade as a mechanism behind increased CBF induced by antipsychotics. Finally, CBF is ultimately a functional marker and this work is important in bridging the considerable gap between the pharmacokinetic and pharmacodynamic effects of compounds with unclear brain functional effects like antipsychotics."} +{"text": "Coxsackievirus B3 (CVB3) is known to induce acute and chronic myocarditis. Most infections are clinically unapparent but some patients suffer from ventricular arrhythmias (VA) and sudden cardiac death (SCD). Studies showed that acute CVB3 infection may cause impaired function of cardiac ion channels, creating a proarrhythmic substrate. However, it is unknown whether low level CVB3+ expression in myocytes may cause altered cardiac electrophysiology leading to VA.Cellular electrophysiology was used to analyze cellular action potentials (APs) and occurrence of afterdepolarizations from isolated cardiomyocytes of wildtype (WT) and transgenic CVB3\u0394VP0 (CVB3+) mice. Further, we studied surface ECGs, monophasic APs, ventricular effective refractory period (VERP) and inducibility of VAs in Langendorff-perfused whole hearts. All used cardiomyocytes and whole hearts originated from male mice.90 with increasing heart rate in Langendorff-perfused hearts. VERP was significantly prolonged in CVB3+ hearts compared to WT . Resting heart rate (HR) in Langendorff-perfused hearts was not significantly different between both genotypes. Electrical pacing protocols induced no VA in WT and CVB3+ hearts.Cellular action potential duration (APD) in WT and CVB3+ myocytes was unchanged. No difference in mean occurrence or amplitude of afterdepolarizations in WT and CVB3+ myocytes was found. Interestingly, resting membrane potential in CVB3+ myocytes was significantly hyperpolarized . Consistently, in Langendorff-perfused hearts, APDs were also not different between WT and CVB3+ whole hearts. Within both groups, we found a heart rate dependent shortening of ADPIn CVB3+ mice, prolonged ventricular refractoriness and hyperpolarized resting membrane potentials in presence of unchanged APD were observed, suggesting that low level CVB3 expression does not promote VA by altered cardiac electrophysiology in this type of chronic myocarditis. These findings may suggest that other mechanisms such as chronic myocardial inflammation or fibrosis may account for arrhythmias observed in patients with chronic enteroviral myocarditis. Within each genotype, we found a heart rate dependent shortening of the ADP90 with increasing heart rate. . In CVB3+ whole hearts, VERP was slight but significantly prolonged compared to WT hearts. .Further, we examined whether CVB3 expression may have an impact on pacemaker activity and resting heart rate. In Langendorff-perfused whole hearts, resting heart rate was not significantly different between both genotypes. . CVB3+ hearts showed only a trend to a minimal higher resting heart rate as WT .Finally, we used electrical pacing protocols to study if CVB3 expression alters cardiac electrophysiology in a proarrhythmic manner leading to increased vulnerability to ventricular arrhythmias in CVB3+ murine hearts. Applied pacing protocols found no enhanced inducibility of ventricular arrhythmias in CVB3+ (n = 5) Langendorff-perfused whole hearts compared to WT (n = 5).The present study was conducted to study the electrophysiological properties in transgenic mice expressing a full length CVB3 cDNA mutant (CVB3\u0394VP0). This mouse model mimics persistence of the CVB3 genome with low levels of CVB3 genome replication and protein expression resembling chronic viral myocarditis or enterovirus-induced dilated cardiomyopathy. In the present study we characterized cellular and whole heart electrophysiology in this CVB3+ mouse model and showed that several cardiac electrophysiological parameters were not altered by CVB3 expression compared to WT. Therefore, generation of arrhythmias in chronic CVB3 myocarditis seems not to depend on altered cardiac electrophysiology but rather on other mechanisms such as chronic inflammation and myocardial fibrosis.2+]i transient and unaltered ICa current density [A previous study by Wessely et al. also investigated restricted viral replication with low level expression of CVB3 proteins in a mouse model . In thes density .90 using stimulation frequencies of 1,2 and 5 Hz in CVB3+ cardiomyocytes compared to WT cardiomyocytes in CVB3+ whole hearts with our Langendorff-setup. We could demonstrated that APD90, in accordance with our cellular data from cardiomyocytes, was not significantly different between CVB3+ and WT whole-hearts and that both genotypes showed preserved heart rate dependent shortening of ADP90 with increasing heart rates was slightly but significantly prolonged in CVB3+ hearts compared to WT hearts . This fiAs known from the clinical setting, patients with advanced stages of heart failure have an increased resting heart rate, which may be due to altered cardiac ion channel properties . Therefof and ICaL [f and ICaL were not impaired by CVB3 expression.Upstroke of the action potential of sinus node pacemaker cells is mainly generated by the funny channel Iand ICaL . As we fPatients with acute and chronic CVB3 myocarditis or cardiomyopathy may suffer from ventricular arrhythmias or sudden cardiac death . Therefo+ current (IKur) with the underlying Kv1.5 channel is responsible for gender differences in cardiac repolarization [Kur current and enhanced Kv1.5 expression with subsequent shortening of ventricular repolarization in male mice compared to female mice [Kur in male C57BL/6 mice was similar to that of female C57BL/6 mice due to the very low testosterone level of the male C57BL/6 mice [Murine studies demonstrated that a stimulatory impact of androgens on the ultra-rapid delayed rectifier Krization \u201339. Andrale mice \u201339. Furtale mice . The whoL/6 mice . In our L/6 mice in male A wide range of patients with CVB3 viral myocarditis are clinically unapparent or present mild symptoms with complete convalescence, whereas a certain proportion of patients suffer from rapid progress and severe cardiac dysfunction due to acute or chronic viral myocarditis leading to cardiomyopathy, ventricular arrhythmias or sudden cardiac death. Our study may imply that in patients with chronic CVB3 myocarditis, electrophysiological properties seem mainly not to be affected by the CVB3 genome in a proarrhythmic manner and thus contribute rather not to arrhythmogenesis. As low level CVB3 gene expression in myocytes can cause cytopathic effects with subXenopus laevis oocytes with acute CVB3 infection found acute alterations of cardiac ion channels IKs, IKr and ICaL which might encourage arrhythmias [Further, our study might contribute to the understanding of differences of acute and chronic CVB3 myocarditis, as a study in hythmias . These fAs our study used a murine model, obtained results cannot uncritically be transferred into the clinical context due to partial different electrophysiological properties between mice and man , 40\u201342. Currently, we can only confirm CVB3+ expression in our homozygous CVB3+ transgenic mice by using nested-PCR without having quantitative data of the CVB3+ expression level, which is subject of a current study. Furthermore, we cannot present imaging data concerning myocardial function of CVB3+ mice, which is also subject of a current study.Our study suggests that low level CVB3 expression does not promote VA by altered cardiac electrophysiology in chronic myocarditis. As previous studies showed that low level CVB3 gene expression in myocytes may lead to cytopathic effects , 18 withOur study in transgenic mice showed that chronic low level CVB3 expression seems not to have a significant impact on cardiac ion channels and cardiac electrophysiology. Our findings may support the hypothesis that other mechanisms like chronic myocardial inflammation over a longer period of time with necrosis and subsequent myocardial scars may predominantly contribute to VAs in chronic CVB3 myocarditis. At the age of mice used in the current study, extent of myocardial inflammation with necrosis and myocardial scars resulting in reduced left ventricular function seems to be less pronounced which may explain that we could not detect VA.In summary, generation of arrhythmias in patients with chronic CVB3-induced cardiomyopathy probably does not rely on altered cardiac electrophysiology but rather on chronic myocardial inflammation over a longer period of time with subsequent necrosis and development of myocardial scars as a proarrhythmic substrate for reentrant arrhythmias. Based on this hypothesis, use of anti-inflammatory and immunosuppressive drugs or angiotensin converting enzyme inhibitors may be promising therapeutic strategies to suppress myocardial remodeling and generation of pro-arrhythmic myocardial scars.S1 TableThe S1 Table contains all original data of the study.(XLSX)Click here for additional data file."} +{"text": "Unfortunately, the original version of this article containeIn the abstract, please replace:4 colony forming units per millilitre (CFU/ml).Patients with clinically suspected VAP undergo BAL and VAP is confirmed by growth of a potential pathogen at [>]10With the following correct version:4 colony forming units per millilitre (CFU/ml).Patients with clinically suspected VAP undergo BAL and VAP is confirmed by growth of a potential pathogen at [\u2265]10In the Interventions, please replace:4 colony forming units per ml (CFU/ml)(13).VAP is confirmed by the widely used threshold of growth of a potential pathogen at [>]10With the following correct version:4 colony forming units per ml (CFU/ml)(13).VAP is confirmed by the widely used threshold of growth of a potential pathogen at [\u2265]10"} +{"text": "Age is a major prognostic factor for malignant gliomas. However, few studies have investigated the management of gliomas in young adults. We determined the role of survival and treatment in young adults with advanced gliomas in a large population from the Chinese Glioma Genome Atlas (CGGA).This study included 726 adults (age \u2265 18) with histologically proven anaplastic glioma or glioblastoma multiforme (GBM). The overall and progression-free survival was determined in young (age < 50) and older groups (age \u2265 50).The study included an older group (OP) of 264 patients and a younger group (YP) of 462patients. In the OP group with GBM and anaplastic glioma, patients treated with RT combined with temozolomide (TMZ) manifested significantly longer OS and PFS compared with patients assigned to RT alone (P < 0.05). In contrast, the YP group diagnosed with anaplastic glioma failed to show any survival advantage with RT plus TMZ compared with RT alone.We observed no survival benefit in young adults (age < 50) with anaplastic glioma when treated with TMZ combined with RT. Our findings warrant further investigation of younger patients diagnosed with anaplastic glioma treated with radiotherapy plus TMZ chemotherapy. Malignant gliomas rank among the most prevalent primary intracranial neoplasms in adults , with anSurgical resection or biopsy, and involved-field radiotherapy are indicated for the treatment of glioblastomas or anaplastic gliomas. Radiation and chemotherapy with temozolomide (TMZ) is the standard of care for patients with glioblastomas.The prognosis of grade II - IV malignant glioma is largely dependent on age. Recent studies have mainly focused on older patients and suggest that the benefit of treatment is reduced with age. Cranial In this study, we summarized the clinical management and evaluated the role of age in clinical outcomes of patients diagnosed with grade III and IV gliomas. We determined the clinical efficacy of treatment across different ages, especially younger adults with advanced gliomas in a large population in the Chinese Glioma Genome Atlas (CGGA).In this study, we analyzed 726 patients diagnosed with advanced (WHO grade III and IV) gliomas from the Chinese Glioma Genome Atlas (CGGA).Patient demographics are listed in Table The standard treatment for malignant gliomas consists of surgery, postoperative radiotherapy, combined with adjuvant TMZ chemotherapy. Surgical resection is the first choice. Gross total resection was conducted in 424 patients (58 %) including 154 (58%) OP and 270 (58%) YP cases, respectively. In the OP group, 131 (50 %) patients were treated with postoperative radiotherapy, and TMZ chemotherapy (RT+TMZ), 36 (14%) underwent postoperative radiotherapy alone (RT), 11 (4%) received postoperative TMZ chemotherapy alone (TMZ) and 25 (9%) were managed with supportive treatment. In the YP group, 221 (48%) were treated with postoperative radiotherapy and TMZ chemotherapy (RT+TMZ), 54 (12%) received postoperative radiotherapy alone (RT), 27 (6%) received postoperative TMZ chemotherapy alone (RT) and 43 (9%) were managed with supportive treatment (Table IDH1) mutations, 1p/19q loss of heterozygosity (LOH) and promoter methylation of O6-methylguanine-DNA methyltransferase (MGMT). The number and frequency of alterations in each age group are listed in Table IDH1 mutations, 34 tumors (5 %) carried LOH on 1p, 12 cases (5 %) showed LOH on 19q, 8 cases (3 %) exhibited 1p/19q co-deletion and 88 tumors (33 %) revealed MGMT promoter methylation. In the YP group, IDH1 mutations were detected in 150 cases (32 %), much higher than in OP (P < 0.0001). The other genetic alterations in the YP group were as follows: LOH involving 1p in 30 cases (6 %), LOH involving 19q in 34 cases (7 %), 1p/19q co-deletions in 28 cases (6 %) and MGMT promoter methylation in 148 tumors (32 %).Patients with adequate tumor specimens were analyzed for genetic changes including isocitrate dehydrogenase 1 (The 243 patients with anaplastic glioma (86%) and 406 patients with GBM (91%) were followed up. Among the two age-specific subgroups with advanced gliomas, the prognosis of YP was more favorable than in OP in terms of overall survival (OS) and progression-free survival (PFS) , patients treated with RT plus TMZ showed significantly longer OS and PFS compared with those treated with RT alone , higher KPS score and MGMT promoter methylation (P = 0.015) were significantly associated with OS in multivariate Cox analysis Figure . A highe) Figure .Advances in radiotherapy , 15 and Since 2005, patients diagnosed with WHO grade III and IV gliomas were recommended TMZ combined with RT. However, patients are unable to afford TMZ therapy due to lack of insurance coverage. We, therIDH1 or IDH2 are significantly younger than those harboring wild-type IDH1 and IDH2.[IDH mutations was low in patients aged above 50 . The NOA-04 trial and validation cohorts in NOA-08 and the German Glioma Network indicated that methylation of MGMT promoter improved outcomes in patients carrying wild-type IDH.[IDH mutations increased the benefit of adjuvant TMZ chemotherapy. A comparison of genetic alterations across different subgroups according to age and treatment, yielded no significant differences in anaplastic gliomas and GBMs. This finding suggests that age was one of the prognostic risk factors in patients treated with adjuvant TMZ chemotherapy for anaplastic glioma.Age is an independent prognostic factor in grade II-IV malignant glioma. Compared with aged and elderly patients, younger adults show a relatively better prognosis as well as improved general condition. They also exhibit greater tolerance to surgical resection. Anaplastic glioma or glioblastoma patients carrying mutant and IDH2., 23 Our type IDH. However,Age, extent of resection and tumor grade are established prognostic factors for gliomas. According to clinical practice guidelines , 26, botOur findings may trigger discussion involving younger adults diagnosed with anaplastic glioma in the combination therapy arm. The positive outcome indicated that RT or TMZ alone was adequate for younger patients diagnosed with anaplastic glioma, which warrants further validation in prospective randomized studiesAdult patients aged at least 18 years and diagnosed with advanced gliomas in the Chinese Glioma Genome Atlas (CGGA) were retrospectively studied. All patients were managed surgically, followed by postoperative radiotherapy, and concomitant and/or adjuvant TMZ chemotherapy at the Glioma Treatment Center of Beijing Tiantan Hospital and Beijing Sanbo Brain Hospital, from October 2004 to July 2012. The study was approved by the hospital ethics committees. All the patients provided written informed consent. The histological diagnosis was validated by two independent neuropathologists and graded according to the 2016 WHO criteria. PatientsStandard care comprised surgery, postoperative adjuvant radiotherapy, and concomitant and adjuvant TMZ chemotherapy. The primary goal of surgery was maximal tumor bulk resection excluding the cortex. MRI findings were used to determine tumor characteristics and the extent of resection within 48 h post-surgery. Abnormal preoperative fluid-attenuated inversion recovery (FLAIR) signals were used to compare the extent of resection based on neuroradiologist reports. The exteTumor tissue samples were resected surgically before starting radio- or chemotherapy. The tissue specimens were snap-frozen and stored in liquid nitrogen until further use.IDH1 mutations , MGMT promoter methylation (DNA pyro-sequencing) and 1p/19q co-deletion (fluorescence in situ hybridization).[Genomic DNA was isolated using the QIAamp DNA Mini Kit (Qiagen). The DNA samples were analyzed using the Nano-Drop ND-1000 spectrophotometer . The sample specimems were analyzed for ization)., 29Survival was monitored clinically during patient visits and via telephone interviews. Patients who were biopsied were excluded from this study. The baseline examination included magnetic resonance imaging (MRI), total blood counts, hematological tests, and physical examinations. During radiotherapy (with or without TMZ), patients were monitored weekly. Comprehensive investigations included physical and radiological assessments 21 to 28 days after radiotherapy and every 3 months subsequently. Adjuvant TMZ therapy included monthly clinical evaluation and comprehensive assessment toward the end of the third and sixth cycles. Tumor progression was defined by a 25% increase in tumor size, new lesions, or an increased need for corticosteroid therapy.SPSS 13.0 software (USA) was used to analyze the data. The differences in clinicopathological characteristics between older and younger adult patients were evaluated with X2 test. OS and PFS were estimated using Kaplan\u2013Meier analysis and compared with two-sided log-rank test. Cox regression analysis was used to assess the prognostic role of the clinicopathological factors and statistically significant treatment protocols based on univariate testing. A P-value < 0.05 was considered statistically significant."} +{"text": "Background: Although small non-coding RNAs are mostly encoded by the nuclear genome, thousands of small non-coding RNAs encoded by the mitochondrial genome, termed as mitosRNAs were recently reported in human, mouse and trout. In this study, we first identified chicken mitosRNAs in breast muscle using small RNA sequencing method and the differential abundance was analyzed between modern pedigree male (PeM) broilers (characterized by rapid growth and large muscle mass) and the foundational Barred Plymouth Rock (BPR) chickens .Methods: Small RNA sequencing was performed with total RNAs extracted from breast muscles of PeM and BPR (n = 6 per group) using the 1 \u00d7 50 bp single end read method of Illumina sequencing. Raw reads were processed by quality assessment, adapter trimming, and alignment to the chicken mitochondrial genome (GenBank Accession: X52392.1) using the NGen program. Further statistical analyses were performed using the JMP Genomics 8. Differentially expressed (DE) mitosRNAs between PeM and BPR were confirmed by quantitative PCR.Results: Totals of 183,416 unique small RNA sequences were identified as potential chicken mitosRNAs. After stringent filtering processes, 117 mitosRNAs showing >100 raw read counts were abundantly produced from all 37 mitochondrial genes (except D-loop region) and the length of mitosRNAs ranged from 22 to 46 nucleotides. Of those, abundance of 44 mitosRNAs were significantly altered in breast muscles of PeM compared to those of BPR: all mitosRNAs were higher in PeM breast except those produced from 16S-rRNA gene. Possibly, the higher mitosRNAs abundance in PeM breast may be due to a higher mitochondrial content compared to BPR. Our data demonstrate that in addition to 37 known mitochondrial genes, the mitochondrial genome also encodes abundant mitosRNAs, that may play an important regulatory role in muscle growth via mitochondrial gene expression control. Production efficiency in animal agriculture is critically important in order to meet the increasing needs of high quality meat protein of a growing global human population. Modern broiler chickens have been highly selected for rapid growth and feed efficiency compared to unselected (progenitor or heritage) chicken breeds. Consequently, the understanding of key regulatory mechanisms for rapid muscle growth and enhanced feed efficiency is imperative with regard to more efficient, and therefore sustainable, food animal production through OXPHOS in most eukaryotic cells were recently discovered in human, mouse and rainbow trout .n = 6 per breed) were humanely killed by cervical dislocation. Breast muscle tissue was rapidly excised and immediately flash frozen in liquid nitrogen. Total RNA was isolated from the tissue using TRIzol reagent according to the manufacturer's instructions. After initial extraction, the RNA samples were treated with DNase I and extracted a second time with TRIzol reagent. Assessing RNA quality using an Agilent 2200 TapeStation instrument revealed that all RNA samples showed >8.0 values of RNA integrity number (RIN). These RNA samples were then used for small RNA sequencing (RNAseq) analysis outlined below.Pedigree male broilers within a single genetic line and RNA extraction were described previously using the NGen program in Lasergene software package . The total mapped counts were transformed to log2 values based on the number of reads per million (RPM) in order to stabilize the variance. The resulting normalized values were then subjected to further statistical analyses using the JMP Genomics 8 . The small RNAs aligned to mitochondrial genome showing >100 average read counts derived by six individual samples, adjusted p-value <0.05 after t-test and log2 fold change >0.5 were considered as differentially expressed (DE) between PeM and BPR. The p-value correction was performed by multiple tests of Benjamini and Hochberg method . The Illumina HiSeq system at this facility used a 1 \u00d7 50 bp single end read technology for sequencing the RNA samples. The RNA sequence FASTQ files that were obtained were mapped to the chicken mitochondrial genome and reverse transcribed to cDNA using an adapter primer containing poly T residues at 3\u2032-end (Table n = 6 for each PeM and BPR). The 2\u2212\u0394\u0394Ct values were calculated using the average \u0394Ct (PeM) \u2212 average \u0394Ct (BPR) indicating that the BPR group values were set to 1 and used for relative quantification by linear fold-change. The statistical significance (p < 0.05) between PeM and BPR values were determined by t-test.Quantitative PCR (qPCR) was conducted following the method reported previously . Ten nanogram of total DNA were subjected to qPCR reaction with primers for mitochondrial genes that included; (a) NADH-ubiquinone oxidoreductase chain (ND): ND1, ND2, ND3, ND4, ND5, ND6, (b) cytochrome oxidase (COX): COX1, COX2, COX3, (c) cytochrome b (CYTB), and (d) ATPase (ATP) 6, and ATP8, as well as nuclear encoded genes using an ABI prism 7500HT system (ThermoFisher Scientific) with PowerUp\u2122 SYBR\u00ae Green Master Mix (ThermoFisher Scientific). The specific oligonucleotide primers are listed in Table n = 6 per group) and sequenced with the 1 \u00d7 50 bp single end read method. A total of ~41 million 50 bp sequence reads were produced with an average of ~3.3 million reads per sample (data not shown). After quality assessment, adapter trimming of raw reads, and alignment to chicken mitochondrial genome (GenBank Accession: X52392.1) using the NGen program, ~1.4 million reads with an average of ~120,000 reads per sample (~350 \u00d7 average coverage) were aligned to the entire sequence of the mitochondrial genome (data not shown). About 97% of reads used in alignment were in the sense orientation of the mitochondrial genome (data not shown). Totally 183,416 unique small RNA sequences were identified as potential chicken mitochondrial genome encoding small RNAs (mitosRNAs) (data not shown). After stringent filtering, 117 mitosRNAs showing >100 raw read counts were abundantly produced from all 37 mitochondrial genes (except D-loop region), all of them were in sense orientation, and the length of mitosRNAs ranged from 22 to 46 nucleotides were derived from the light strand while 113 mitosRNAs were derived from heavy strand mitochondrial DNA <0.05 were identified. All of the DE mitosRNAs were upregulated in PeM breast, except those that were encoded by the 16S-rRNA gene region . Since DE mitosRNAs were mostly found in protein coding genes (mRNA), the DE mitosRNAs were compared to gene expression derived from the RNAseq analysis conducted on the same muscle samples .The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer DC and handling Editor declared their shared affiliation."} +{"text": "Conversely, the sensitivity of ECV for the detection of focal disorders is unclear. This may be important for subendocardial disease since the standard methodology for ECV requires that \"regions of interest.. have adequate margins of separation from tissue interfaces prone to partial volume averaging such as between myocardium and blood\" [Flow-Independent Dark-blood DeLayed Enhancement technique (FIDDLE) that increases the conspicuity of subendocardial hyperenhancement, by making the blood pool black [Delineation of diseased and normal tissue is fundamental to identifying cardiac pathology. Studies suggest that parametric extracellular volume fraction (ECV) imaging is superior to conventional delayed enhancement for detection of diffuse, d blood\" . We haveol black . In thisCanines (n = 7) underwent variable coronary occlusion to create a range of infarct sizes including 2 sham operated controls. CMR was performed 2-5 days after MI followed immediately by excision and TTC staining to provide a gold standard histopathology reference. ECV was calculated using T1-maps generated with MOLLI sequence (SIEMENS WIP 448B) before and after gadolinium administration (0.2 mmol/kg). FIDDLE images were acquired in identical slice locations 10-20 minutes after gadolinium administration. FIDDLE and ECV analysis were performed separately, blinded to subject identity and pathology. ECV was calculated as previously reported . 26 patiPathology confirmed MI was found only in animals that underwent coronary occlusion. Mean ECV in controls was 29.8% \u00b1 2.2%, and abnormal ECV was defined as >2SD above the mean (34.2%). Analysis of the 41 matched slices in canines showed that ECV was insensitive for the detection of subendocardial infarcts (3%) while transmural infarcts were routinely detected (91%). Conversely, FIDDLE had high sensitivity for the detection of both subendocardial (93%) and transmural (100%) infarcts. In patients, mean ECV in controls was 26.3% \u00b1 2.6%, and abnormal ECV was . Using FIDDLE as the reference standard, ECV was insensitive for the detection of subendocardial MI (26%). Figure FIDDLE provides excellent visualization and detection of infarcts, while ECV frequently misses subendocardial infarcts. This suggests that ECV imaging is not optimal for the detection of focal, subendocardial disease."} +{"text": "Measurements of myocardial extracellular volume (ECV) through cardiac magnetic resonance (CMR) based T1 mapping allow for non-invasive quantification of diffuse myocardial fibrosis. The evaluation of the right ventricle (RV), often mainly affected in congenital heart disease (CHD), is complicated by its typical morphologic characteristics - a thin myocardium and a distinct trabeculation. This study aims to evaluate ECV measurements of the RV in patients with CHD and to introduce a tool that simplifies RV ECV analysis.CHD patients were prospectively enrolled and compared with 17 healthy volunteers (mean age 25.1 \u00b1 2.7 years). T1 maps were generated with Modified Look-Locker Inversion recovery (MOLLI) T1 mapping in a single midventricular plane in short axis (SAX) and transverse orientation (TRANS) before and 15 minutes after bolus application of Gd-DOTA or Gd-DTPA. To measure ECV, T1 values of the anterior or inferior RV wall were used. Regions of interest (ROIs) were evaluated with regard to image quality and ROI thickness . ECV from plane RV ROIs were compared with ECV obtained from a custom-made tool that derives the mean T1 values from a curved line manually drawn in the center of the myocardial wall (\"centerline ECV\").RV ECV could be determined in 40 patients (32 \u00b1 0.05) and 9 volunteers (0.28 \u00b1 0.03) with a strong correlation of ECV values from SAX and TRANS . In both groups average image quality was rated with grade 1 and mean RV ROI thickness with grade 2. ECV could not be measured in cases of insufficient contrast between blood and myocardium or ROI thickness < 1 pixel. ECV from ROIs and corresponding centerline ECV correlated strongly in SAX and TRANS in CHD patients and volunteers .1. RV ECV can be assessed with T1 mapping in SAX and TRANS provided thata) image quality allows for sufficient distinction between blood and myocardium, andb) RV wall thickness is > 1 pixel.2. The application of a simple line centered in the RV myocardium instead of a plane ROI simplifies and accelerates measurements of RV ECV.Under these conditions, measurements of RV ECV can be integrated into clinical CMR routine across a wide spectrum of CHD."} +{"text": "Virgin Islands Department of Health (USVI DOH) was notified by a local ophthalmologist of an unexpected increase in the number of patients with suspected epidemic keratoconjunctivitis (EKC) during the preceding month. EKC is a severe form of acute conjunctivitis caused by human adenoviruses (HAdVs). Clinical illness typically lasts 1 to 3 weeks and is usually self-limited; treatment is supportive (1). HAdVs can survive for weeks in the environment and are resistant to common disinfectants (A case of EKC was defined as 1) a diagnosis by an ophthalmologist or optometrist of EKC, adenoviral conjunctivitis, or viral conjunctivitis (excluding conjunctivitis diagnosed in association with presumed Zika virus infection); or 2) laboratory confirmation of HAdV type 8 (HAdV-8) from a specimen collected by conjunctival swab in a person on the affected island during June 1\u2013November 29, 2016. A health care\u2013associated case, was defined as a case in a person who had visited an eye care practice \u226414 days before onset of symptoms.Available medical records were reviewed for patients with diagnoses of acute conjunctivitis during June 1\u2013November 4, 2016 from all six eye care practices on the affected island. Additional cases were identified prospectively through collection of conjunctival swabs from patients evaluated for acute conjunctivitis at eye care practices, a hospital emergency department, and two family practice clinics during October 14\u2013November 29, 2016.Environmental testing was conducted, and routine infection control practices were assessed at the two eye care practices where health care\u2013associated transmission was suspected to have occurred: the initial reporting practice (practice A) and a second eye care practice (practice B). Environmental samples were collected from eye care equipment and high-touch surfaces in waiting areas and patient examination rooms. Conjunctival and environmental swabs were tested at CDC using HAdV real-time polymerase chain reaction (qPCR). Positive specimens were molecularly typed, based on sequencing of hexon hypervariable regions 1\u20136 and inoculated into A549 cells for virus isolation. Whole genome sequencing was performed on five selected cell culture isolates, obtained from patients who were infected at the beginning, middle, and end of the outbreak.Seventy-eight cases were identified in patients from four eye care practices, two family practices, and the hospital emergency department. The median patient age was 45 years (range = 9 months\u201390 years), and 33 (42%) were men. Ocular signs and symptoms included redness (68%), watery discharge (50%), and pain (29%). Severe signs included corneal infiltrates (17%) and pseudomembranes (6%). At least 12 cases (15%) were health care\u2013associated . One hea,Infection prevention and control guidance was provided to six eye care practices and the emergency department. Recommendations included 1) maintaining proper hand hygiene, 2) cohorting patients with suspected EKC in the health care setting, 3) refraining from using contents from eye drop bottles for more than one patient, 4) using an Environmental Protection Agency\u2013registered disinfectant with proven activity against HAdVs to decontaminate surfaces and equipment, and 5) furloughing symptomatic employees ,. PatientHealth care\u2013associated transmission of EKC in this outbreak highlights the importance of infection control in eye care practices, including the use of disinfectants with proven efficacy against adenoviruses. The occurrence of household transmission also underscores the importance of patient education regarding measures to prevent the spread of EKC."} +{"text": "Pi3kca and/or Pten mutations detected in 11 of 13 (85%) long latency cases . Eighty-two percent (9/11) of tumors carried the Pik3ca H1047L/R hot-spot mutation, as frequently found in human breast cancer. These tumors were luminal-like and mostly ER/PR+, as in humans. Transcriptome profiling indicated a significant activation of the PI3K-Akt pathway (p=3.82e-6). On the other hand MPA+DMBA induced short latency tumors displayed mutations in cancer drivers not commonly found mutated in human breast cancer (e.g. Hras and Apc). These tumors were mostly basal-like and MPA exposure led to Rankl overexpression (60 fold induction) and immunosuppressive gene expression signatures. In summary, long latency DMBA induced mouse mammary tumors reproduce the molecular profile of human luminal breast carcinomas representing an excellent preclinical model for the testing of PIK3CA/Akt/mTOR pathway inhibitory therapies and a good platform for the developing of additional preclinical tools such as syngeneic transplants in immunocompetent hosts.Controversy always existed on the utility of chemically induced mouse mammary carcinogenesis models as valid equivalents for the study of human breast cancer. Here, we performed whole exome and RNA sequencing on long latency mammary tumors (218 \u00b1 27 days) induced by the carcinogen 7,12-Dimethylbenzathracene (DMBA) and short latency tumors (65 \u00b1 11 days) induced by the progestin Medroxyprogesterone Acetate (MPA) plus DMBA in CD2F1 mice. Long latency tumors displayed a high frequency of Chemically induced rodent models of mammary cancer have been extensively used over the years to emulate human breast carcinogenesis. All models in mice and rats have specific advantages and limitations. Mammary tumors can be induced in susceptible rat strains after single doses of carcinogens such as DMBA or nitrosomethylurea (NMU). Rat tumors are not extremely invasive beyond the mammary fat pad, have short latency, seldom metastasize and are highly hormone-dependent and for that reason were widely used as models of estrogen dependent breast cancers . On the Hras oncogene in NMU induced rat tumors [In terms of the molecular profiles of chemically induced mammary tumors in rodents, not much has been learned beyond the early work of Barbacid and coworkers demonstrating the signature activation of the t tumors , 5. Hrast tumors \u20138. SinceTo the best of our knowledge no study to date has analyzed comprehensively the mutational profile of chemically induced mouse mammary tumors in order to understand how closely these models reproduce human breast cancer molecular profiles. To this end we comprehensively analyzed the mutational and transcriptomic profile of DMBA and MPA-DMBA induced mouse mammary tumors by means of exome and RNA sequencing.Exome-Seq data on 22 mouse mammary tumors indicated a median of 75% targeted genome loci having at least 40X coverage. Overall we identified 25898 single base substitutions including 18104 non-synonymous single nucleotide variants (SNVs), 6068 synonymous, 1674 stop-gain and 52 stop-loss SNVs . We alsoAlmost 64% of the SNVs were A>T:T>A transversions followed by 16% of G>T:C>A substitutions. These data are in agreement with previous studies demonstrating that A>T:T>A and G>T:C>A transversions were the most predominant types of base pair substitutions (44% and 24% respectively) induced by DMBA in rat mammary tissue . MutatioThe total mutation rate across tumors was 21.7 mutations per Mbp on average with a range of 9.8 to 57.7 mutations per Mbp, indicating that some mouse mammary tumors have significantly higher mutation rates than others. In this sense, MPA+Dx4 (see Methods for nomenclature of treatment groups) derived tumors showed a statistically significant increased rate of mutations/Mbp compared with Dx4 and MPA+Dx2 derived tumors , Pik3ca in 11 of 22 mammary tumors (50%), Hras in 7 of 22 (32%), Mll3 in 7 of 22 (32%) and others such as Nim, Atrx, and Ptprc in 5 of 22 each one (i.e. 23% of cases mutated for each gene) and Apc, Trp53, Irf4 and Col2a1 were found mutated in 18% of cases each one (4 of 22 of cases mutated for each gene). Interestingly, we identified a very distinct pattern on the mutational profile of cancer drivers genes clearly differentiating tumors that develop with short latency from those with long latency. The short latency tumors (< 100 days for tumor development) produced mostly by the MPA+Dx4 protocol, were characterized by the predominant presence of mutations affecting the Hras oncogene in 6 of 9 (67%) of such early tumors and only one case displaying an Hras mutation Figure .Unsupervised analysis of RNA-Seq data demonstrates a clear segregation of normal CD2F1 mouse mammary samples and the chemically induced mammary tumors Figure . In addiRankl transcript was the most up-regulated gene in MPA treated normal and tumor samples microenvironment that facilitates the expansion of the highly proliferative cells ultimately leading to the rapid development of mammary tumors (i.e. short-latency tumors).Taken together, the present study suggests that MPA treatment accelerates mammary tumorigenesis in this and other rodent mammary tumor models via Pik3ca and/or Pten mutations in 64% (14 of 22) of the mammary tumors analyzed. However, we observed that long latency tumors , contrasting with the short latency tumors characterized by carrying Hras driver mutations (67%) Figure .PIK3CA and PTEN mutations were thought to be mutually exclusive in various cancers with the exception being endometrial tumors where coexistent mutations of both genes were reported [PIK3CA mutation and PTEN protein loss in breast carcinomas showed that both alterations were indeed not mutually exclusive indicating that PIK3CA mutations coexist with PTEN loss in ER + breast cancers [Pik3ca and Pten behave as co-occurring mutations in the chemically induced mouse mammary tumors analyzed and frameshift insertion (1 of 12) resulting in stop-gain mutations, the remaining cases (6 of 12) consist in deleterious Pten mutations leading to loss-of-function as was predicted by the SIFT algorithm.Fifty percent of the tumors with Pik3ca gene in our chemically induced mouse mammary tumors and in human invasive breast carcinomas [Pik3ca (H1047L/R), which is by far the most frequent PIK3CA mutation reported in human invasive breast carcinomas . Furthermore, the H1047R mutation was demonstrated to activate PI3K signaling in human mammary epithelial cells and induces tumor formation in nude mice [The comparative frequency of mutations on the al.org/) is shownude mice .PIK3CA somatic mutations are detected in over 34% of human breast cancers and there is a need for pre-clinical mouse mammary cancer models that more closely reproduce the histology and biology of the human counterparts for the in vivo testing of anti-PIK3CA therapies [reviewed in We consider this a very significant finding considering that Pik3r3 over-expression, Pik3ca and Pten mutations) and multiple downstream effectors such as Ccnd1, Cdk4, ATF4 and Myb among others.RNA-Seq analysis revealed a group of 3675 differentially expressed genes was the top up-regulated gene in Dx4 induced tumors . OPN is a multi-functional cytokine that binds to cell surface receptors, such as CD44 and integrins, which are involved in immune response, cell survival, adhesion and migration. OPN has been previously associated with tumor formation, progression and breast cancer metastasis [e.g.: Itgb4/6 and Itga3/6/8) were also up-modulated in Dx4 chemically induced tumors. Together, our results suggest that the overexpresion of OPN, several PI3K-Akt related binding receptor and several down-stream modulators including Pik3r3, Ccnd1 and Cdk4 among others could be part of a positive autoregulatory loop controlling tumor growth in long latency chemically induced tumors.In this sense, tastasis . Recentltastasis . It is iHras oncogene. Additionally, transcriptomic analysis suggests that MPA exposure accelerates mammary tumorigenesis not only via Rankl-mediated proliferative expansion of the carcinogen \u2018initiated\u2019 mammary epithelial cells carrying cancer driver mutations such as Hras activating and Apc loss of function mutations, but also by generating an immunosuppressive tumor microenvironment that likely facilitates the development of short-latency basal-like mammary tumors. On the other hand, DMBA-induced long-latency tumors were characterized by A>T and G>T substitutions frequently affecting the Pik3ca and Pten cancer driver genes. More importantly, 82% of the cases with Pik3ca missense mutations displayed the characteristic kinase domain H1047L/R \u2018hot-spot\u2019 activating mutation that has been reported as the most frequent PIK3CA mutation in human breast cancers. Furthermore, transcriptomic analysis of the DMBA induced tumors pointed to PI3K-Akt signaling pathway activation as extremely common in long latency luminal-like tumors.The comprehensive molecular characterization of chemically induced mouse mammary tumors allowed us to define mutational and transcriptomic signatures driving the DMBA and MPA induced carcinogenic processes. Short latency tumors derived mostly by the MPA+Dx4 protocol, were characterized by high mutational burden affecting multiple cancer driver genes including the predominant frequency of mutations affecting the PIK3CA mutated counterparts. Thus, the DMBA induced mouse mammary tumor model appears as an excellent system to provide a better understanding of Pik3ca/Pten mutant driven mammary cancer pathogenesis but also as a prime candidate for further developing pre-clinical models (e.g. syngeneic transplant lines) for the in vivo testing of Pik3ca/Akt/mTor pathway inhibitory therapies.As recently reviewed , there iThirty fresh-frozen CD2F1 (BALB/c x DBA/2) mouse mammary normal and tumor samples were obtained from a tumor bank at the Aldaz lab from studies performed and reported 20 years ago in 1996 . BrieflyRoutine histopathologic evaluation was performed on all tumors by means of formalin fixation, paraffin embedding and hematoxylin and eosin stained sections. For tumor classification we followed the recommendations of the Annapolis consensus report for the analysis of mouse mammary tumors Supplem. All samhttp://picard.sourceforge.net/). Tumors were called against normal DNA isolated from two wild-type CD2F1 mice. Subsequently, single-nucleotide variants were identified using MuTect v1.1.4 [http://www.ensembl.org/Tools/VEP) to compute the SIFT scores and VarScan software (http://dkoboldt.github.io/varscan/) was employed for indels detection. Detected somatic mutations were further filtered by focusing only on those genes considered cancer driver genes as defined by the COSMIC Cancer Gene Census database (http://cancer.sanger.ac.uk/census/). The Comet Exact Test (CoMEt) algorithm was employed to identify mutually exclusive mutations [DNA from 22 mouse mammary tumor samples and two normal mouse mammary samples from strain CD2F1 were purified using the DNeasy Blood and Tissue Kit (Qiagen). Only DNA samples with 260/280 ratios greater than 2.0 were processed for library construction using the SPRIworks Fragment Library Kit I (Beckman Coulter). Four libraries were pooled together and processed for exome capture using the NimbleGen SeqCap EZ Developer Library , covering ~ 54.3 Mbp of target sequence. 76 nt paired-end sequencing was performed using an Illumina HiSeq2000 platform at our Department's NGS Facility. Image analysis, base-calling, and error calibration were performed using Illumina's Genome analysis pipeline. Sequencing was performed reaching an average depth of 40X per sample. Sequenced 76 bp tags were aligned against the mouse reference genome (GRCm38/mm10) using BWA v0.7.3 and marked for duplicates using Picard v1.88 and the InnateDB resource (http://www.innatedb.com/) based on the list of deregulated transcripts between normal and mouse mammary tumors . Data integration and visualization of differentially expressed transcripts were done with R/Bioconductor [RNA was isolated and purified using TRIzol reagent (Life Technologies) and RNeasy mini kit (Qiagen). RNA concentration and integrity were measured on an Agilent 2100 Bioanalyzer (Agilent Technologies). Only RNA samples with RNA integrity values (RIN) over 8.0 were considered for subsequent analysis. mRNA from normal mouse mammary and tumor samples were processed for directional mRNA-seq library construction using the ScriptSeq v2 RNA-Seq Library Preparation Kit (Epicentre) according to the manufacturer's protocol. We performed 76 nt paired-end sequencing using an Illumina HiSeq2000 platform and obtained ~23-37 million tags per sample. The short sequenced reads were mapped to the mouse reference genome (mm10) by the splice junction aligner TopHat V2.0.10 . We emplonductor and the onductor . In addionductor ."} +{"text": "Journal of Global Oncology, Al-Wassia et al1 reported on the outcomes of Saudi Arabian patients with nasopharyngeal cancer treated with primarily neoadjuvant chemotherapy followed by concurrent chemoradiotherapy. We have a few questions about the study.In their recent article in 2 clearly showed no benefit? What were the doses of the docetaxel, cisplatin, 5-fluorouracil regimen used as neoadjuvant therapy? How were the patients selected for cisplatin either 100 mg/m2 every 3 weeks or 30 mg/m2 once per week? Did patients receive adjuvant chemotherapy?Why was neoadjuvant chemotherapy given when the meta-analysis of chemotherapy in nasopharyngeal cancer On what basis were patients selected for neoadjuvant chemotherapy followed by concurrent chemoradiotherapy, neoadjuvant chemotherapy followed by radiotherapy, or concurrent chemoradiotherapy alone? Did any patients undergo resection of residual neck nodes after completion of therapy? How were the patients with relapse managed?Finally, was Epstein-Barr virus testing performed and was there any association with tobacco or alcohol consumption?"} +{"text": "Chikungunya virus causes fever and severe polyarthritis or arthralgia and is associated with neurologic manifestations that are sometimes challenging to diagnose. We demonstrate intrathecal synthesis of chikungunya antibodies in a patient with a history of acute infection complicated by encephalitis. The specificity of the intracerebral immune response supports early chikungunya-associated encephalitis diagnosis. Aedes mosquitoes (Chikungunya virus (CHIKV) is an alphavirus transmitted by infected In January 2016, a 69-year-old woman had sudden fever (38\u00b0C), intense generalized arthritis, prostration, and cognitive alterations characterized by forgetting and exchanging words. She used analgesics without relief. After 1 week, a maculopapular rash with intense pruritus appeared on her upper limbs. A few days later, her rash and fever abated, but other symptoms continued. She was referred for consultation 3 months after symptom onset. Physical examination revealed bilateral finger and knee arthritis Figure. Euroimmun, Luebeck, Germany); Panbio Dengue IgG Indirect ELISA and Dengue IgM Capture ELISA ; and immunochromatographic assay . Results for specific IgG and IgM are shown in the 3; protein 29 mg/dL; glucose 40 mg/dL; blood\u2013CSF barrier function based on albumin quotient 6 \u00d7 10\u22123; IgG index 0.43 (reference <0.7); and intrathecal IgG fraction (IgGIF) of total IgG found in CSF, <0% (IF <0%). The sample dilutions for dengue were 1:8 for CSF and 1:4,000 for serum. Sample dilutions for CHIKV were 1:2 for CSF and 1:101 for serum with progressive reduction for another 2 months.Our results demonstrate the quantitation of intrathecally synthesized CHIKV IgG. We diagnosed CHIKV-associated encephalitis on the basis of fever and altered mental status for >24 hours and positive CHIKV IgM antibodies in serum (The identification of specific etiologies of viral encephalitis may be difficult in arbovirus-endemic areas (CHIKV has attracted increasing attention because of its spatial spread and the high number of epidemics. Chikungunya has been associated with debilitating arthropathy for months or years after the initial infection, along with severe neurologic complications such as encephalitis (Additional information about chikungunya virus infection in a patient in Brazil."} +{"text": "S1 FigTenebrio molitor (Ts), Alphitobius diaperinus (As) and Hermetia illucens (Hs). A similar gel was stained with Coomassie (right).Native PAGE stained with 3 mM L-DOPA (left) showed no active bands for extracts treated with sodium bisulfite from (TIF)Click here for additional data file."} +{"text": "Cunningham et al. demonstrate that the Id family of helix\u2013loop\u2013helix proteins is both necessary and sufficient to direct cardiogenic mesoderm formation from frog embryos to human embryonic stem cells. Deciphering the fundamental mechanisms controlling cardiac specification is critical for our understanding of how heart formation is initiated during embryonic development and for applying stem cell biology to regenerative medicine and disease modeling. Using systematic and unbiased functional screening approaches, we discovered that the Id family of helix\u2013loop\u2013helix proteins is both necessary and sufficient to direct cardiac mesoderm formation in frog embryos and human embryonic stem cells. Mechanistically, Id proteins specify cardiac cell fate by repressing two inhibitors of cardiogenic mesoderm formation\u2014Tcf3 and Foxa2\u2014and activating inducers Evx1, Grrp1, and Mesp1. Most importantly, CRISPR/Cas9-mediated ablation of the entire Id (Id1\u20134) family in mouse embryos leads to failure of anterior cardiac progenitor specification and the development of heartless embryos. Thus, Id proteins play a central and evolutionarily conserved role during heart formation and provide a novel means to efficiently produce cardiovascular progenitors for regenerative medicine and drug discovery applications. Heart formation begins during gastrulation with the specification of cardiogenic mesoderm progenitors (CMPs) that migrate anteriorly to form the cardiac primordium that assembles into the fully formed heart . IntenseMesp1 and Mesp2 and cellular migration (Prickle1 and RasGRP3) while actively repressing genes regulating pluripotency and early mesoderm (T) and endoderm (Foxa2 and Sox17) fates transcription factors nd Mesp2 under th2 (Eomes regulate7) fates . Althouganalysis and in vanalysis have shoId1, a HLH transcriptional regulator, as a single factor sufficient to control the emergence of Kdr+ CMPs both in mouse and human embryonic stem cells . Mechanistically, we discovered that Id proteins mediate their evolutionarily conserved role by activating the expression of agonists of cardiogenic mesoderm formation while inhibiting antagonists\u2019 activity (Tcf3 and Foxa2). Finally, CRISPR/Cas9-mediated deletion of all four Id family members in mouse embryos blocked early cardiac progenitor formation and yielded embryos without a heart. The heartless phenotype was unique to the quadruple knockout, indicating compensatory or redundant functions of the Id proteins in the formation of the cardiac mesoderm. These findings reveal an unexpected role for Id proteins as the earliest determinants of cardiac cell fate in vertebrates.We reported recently that attenuating Acvr1b signaling in mesendoderm segregates cardiogenic mesoderm from endoderm, whereas persistent Acvr1b signaling drives cells to form endoderm . Thus, w+, Foxa2+, and T+) at days 3\u20134 of differentiation in response to Activin/Nodal signaling and subsequently differentiate into either Foxa2+ definitive endoderm or Kdr+ cardiogenic mesoderm (diagrammed in let-7 or miR-18 mimics or siRNAs directed against their respective mRNA targets: Acvr1b or Smad2 (day 3) transfection (day 4). Microarray data revealed 33 genes that were up-regulated in response to Acvr1b siRNA relative to a scrambled sequence siRNA control, of which 14 were confirmed by quantitative PCR (qPCR) , inhibitors of bHLH transcription factors (Id1 and Id3), and a mediator of DNA demethylation (Gadd45g). Of the six remaining candidates, three are signaling pathway modulators , two are involved in RNA processing (Elavl3 and Tnrc6a), and one encodes a protein with two centrosome-associated domains but no known function (Grrp1). Interestingly, none of the 14 candidates were shown previously to directly control cardiogenic mesoderm formation, suggesting that we identified a novel molecular signature marking differentiating CMPs.To identify the downstream effectors of cardiogenic mesoderm formation, we analyzed mRNA expression 24 h after R (qPCR) E. ConsisKdr-eGFP reporter system and the anterior lateral mesoderm where specified cardiac precursors are located . The cell lines were treated with Activin A (but not with Acvr1b siRNA), and the resulting differentiation was assessed on day 6. Id1 was sufficient to massively direct ESCs to differentiate toward the Kdr+ mesoderm without attenuating Acvr1b expression , while the other genes had less potent effects . Quantitatively, the conversion rate of Id1-overexpressing mESCs into Kdr-eGFP+ mesoderm was \u223c60% as compared with only 3.65% for control ESCs hESC line that stably overexpresses mouse Id1, since the HLH domains are identical across these species G,H. Cons hESCId1 I,J.+/KDR+ mesoderm progenitors (iMPs) was consistently preceded by the up-regulation of Evx1/EVX1 and Grrp1/GRRP1 (days 3/4 in mESCs ), transferred into M2 medium (Millipore), and injected with the Cas9 mRNA/sgRNA solution into the cytoplasm. Injected embryos were then reimplanted into recipient pseudopregnant ICR female mice. Implanted females were sacrificed 8\u20139 d after reimplantation; yolk sac DNA was collected for genotyping by PCR (Bioline MyTaq extract kit) followed by DNA deep sequencing (Ilumina Nextera kit for library preparation and Illumina HiSeq 1500 for sequencing). Sequences were analyzed, and variant alleles were recorded using Integrative Genomics Viewer (IGV) genome browser (Broad Institute). For off-target analysis, the top eight off-target sites were identified using the tool at http://crispr.mit.edu; these regions were PCR-amplified and Sanger-sequenced.In order to generate Id1\u20134 mutant embryos, eight sgRNAs were designed to target sites near the ATG translation initiation site and near the beginning of the HLH domain for each Id gene (see the t-test (P < 0.05).Each experiment represents at least quadruplicate biological replicates per condition. Statistical analysis was performed with unpaired Student's"} +{"text": "Late gadolinium enhancement (LGE) using magnitude inversion recovery (IR) or phase sensitive inversion recovery (PSIR) has become clinical standard for assessment of myocardial scar. However, there is no clinical standard for quantification of myocardial scar even though multiple methods have been proposed . Simple The new automatic algorithm was implemented using an intensity threshold defined by expectation maximization (EM) followed by the weighted approach to take partial volume effects into account and improved detection of microvascular obstruction.The new automatic algorithm and reference delineation in IR and PSIR images was validated against TTC in six pigs with myocardial infarction imaged after seven days of reperfusion. The new automatic algorithm was also validated against reference delineation in 127 patients from the multi-center, multi-vendor studies CHILL-MI and MITOCARE in IR (n = 75) and PSIR (n = 52) images. All patients underwent CMR imaging within 2-6 days following first time ST-elevation myocardial infarction (STEMI) treated with percutaneous coronary intervention (PCI). Reference delineation was performed by a core lab using the original weighted algorithm followed by manual corrections and consensus reading. Analysis was performed using bias (mean \u00b1 standard deviation) and linear regression analysis (correlation coefficient). Results are expressed as percent left ventricular mass %LVM.Infarct size by TTC was 9 \u00b1 6 %LVM. Bias to TTC for the new automatic algorithm was -1 \u00b1 1 %LVM and -2 \u00b1 2 %LVM in IR and PSIR images, respectively (Table The new automatic algorithm was validated against TTC and in multi-center, multi-vendor patient data with a low bias for both IR and PSIR images. Results show that the new algorithm performs equally well in images acquired with both sequences. The new automatic algorithm can be used as an improved tool for segmentation of myocardial scar in IR and PSIR images."} +{"text": "The internet and social media provide an unprecedented opportunity to transform early psychosis intervention services. This study aimed to capture concerning patterns of social media activity associated with the onset and persistence of psychotic symptoms.Facebook and Twitter archives were extracted from over 150 participants with psychotic disorders, mood disorders and healthy controls. Machine learning was used to build classifiers aiming to identify patterns and distinguish between groups.Linguistic analysis of Twitter commentary identified significantly increased use of interpersonal pronouns (p < 0.001), decreased emphasis on friendship (p < 0.001) and increased emphasis on health (p < 0.001) in individuals with psychosis. Preliminary classifiers correctly recognized participants with psychotic disorders (n=62) from healthy controls (n=24) with an average accuracy of 80% and distinguished participants with psychosis from those with mood disorders (n=39) with an average accuracy of 70%. Further analysis identified shifts in language use of participants with psychosis who experience a relapse (n=18) including significant increases in the use of swearing (p<0.05), first-person pronouns (p<0.05) and negations (p<0.05). We additionally identified significant differences in the profile pictures (p<0.005) and structure of messages posted (p<0.005) by youth with psychosis who experienced a psychotic relapse.Identifying markers in social media activity associated with worsening psychotic symptoms offers the prospect that social media may be a clinically useful tool to identify patients in the earliest phases of relapse."} +{"text": "Background/Purpose. Understanding the practices of pediatric rheumatologists in diagnosing and treating chronic nonbacterial osteomyelitis (CNO) can provide important information to guide the development of consensus treatment plans. The objectives of this study were to determine physicians' approaches to (1) diagnosing and monitoring CNO, (2) ordering a bone biopsy, and (3) making treatment decisions. Methods. A survey was distributed among members of the Childhood Arthritis and Rheumatology Research Alliance using a web-based questionnaire. Results. 121 of 277 (41%) attending physician members completed the survey. Plain radiographs (89%) were most commonly used followed by regional MRI (78%), bone scintigraphy (43%), and whole-body MRI (36%). The top three reasons for performing a biopsy were constitutional findings (66%), unifocal bone lesions (64%), and nocturnal bone pain (45%). Nearly all responders (95%) prescribed nonsteroidal anti-inflammatory drugs (NSAIDs) as initial therapy. For patients who failed NSAID treatment, methotrexate (67%), tumor necrosis factor inhibitors (65%), and bisphosphonates (46%) were the next most commonly used treatments. The presence of a spinal lesion increased the use of bisphosphonate treatment. Conclusion. The diagnostic approach and disease activity monitoring for CNO varied among surveyed physicians. Our survey findings provided important background for the development of consensus treatment plans for CNO. Chronic nonbacterial osteomyelitis (CNO), also known as chronic recurrent multifocal osteomyelitis (CRMO), is an autoinflammatory bone disease of unknown cause that can result in persistent bone pain, bone destruction, functional disability, and pathological fractures. The diagnosis of CNO is based on a history of bone pain, findings of bony tenderness with or without swelling, imaging confirmation of a lytic and/or sclerotic bone lesion, and/or bone edema with negative findings for malignancy and infection of the affected area \u20135. At diImaging plays an essential role in evaluating children with CNO. Plain radiographs are readily available but not sensitive , 11. TypDue to the rarity of the disease and the We conducted a survey of pediatric rheumatologists through Childhood Arthritis and Rheumatology Research Alliance (CARRA). CARRA is a North American organization of more than 425 pediatric rheumatologists, researchers, and research coordinators who are working together to advance the health and quality of life of children living with pediatric rheumatic diseases. Consensus treatment plans for systemic juvenile idiopathic arthritis (JIA) , polyartThe objectives of this study were to determine (1) physicians' approaches in diagnosing and monitoring disease activity, (2) which disease features physicians consider important for ordering a bone biopsy, and (3) physicians' treatment choices.https://doi.org/10.1155/2017/7694942) was administered through a REDCap database [t-tests of NSAID versus each of four other drug classes: glucocorticoid, disease modifying antirheumatic drug (DMARD), biologic, and bisphosphonate. Tests were adjusted for multiple comparisons using the Bonferroni correction.This research project was approved by the Seattle Children's Hospital Institution Review Board. The survey was developed based upon feedback from members of the CARRA Scleroderma, Vasculitis, and Rare Diseases (SVRD) subcommittee. Multiple-choice questions were used and case scenarios were presented in addition to general questions. The survey (One hundred and twenty-one of 277 (41%) attending physician members and 18 of 83 trainee physicians from CARRA answered the survey. Only responses from attending physicians who have been involved in care of patients with CNO were retained in the final analytic dataset (n = 109) . Sixty-sn = 109) but did n = 109) .Reported frequencies of bone biopsy were never 0%, rarely 12%, sometimes 28%, often 39%, or always 21%. The top three reasons for performing a biopsy were constitutional changes such as fever, weight loss, night sweats (66%), unifocal bone lesion (64%), and nocturnal bone pain (45%). The top three reasons for not performing a biopsy were involvement of \u201ctypical sites\u201d (64%), the presence of multiple bone lesions (61%), and CNO-associated conditions such as psoriasis, inflammatory bowel disease (IBD), or enthesitis-related arthritis (ERA) (50%). \u201cTypical sites\u201d were identified as the clavicle (54%), metaphysis of long bones (36%), vertebral bodies (28%), long bones in lower extremity (27%), mandible (21%), long bones in upper extremity (15%), epiphysis of long bones (6%), diaphysis of long bones (5%), other (7%) including pelvis, and sternum. The reasons for obtaining or not obtaining a bone biopsy were similar regardless of whether the physician reported ordering biopsies rarely or often (data not shown).The top three ways of identifying bone biopsy site were ease of access (65%), leaving the decision to the orthopedic surgeon/interventional radiologist (64%), and the presence of a lytic lesion on X-ray (30%).The following histologic features were considered to be indicative of CNO: presence of plasma cells, macrophages, and neutrophils (69%), reparative changes such as fibrosis (51%), normal bone (20%), signs of necrosis (19%), unclear reason (15%), biopsy rarely obtained (12%), and other (6%).Among all imaging modalities used often or always, X-rays (89%) were most commonly used diagnostic imaging modality, followed by regional MRI (78%), and bone scintigraphy (43%) . Fifty-on = 39), 59% found the short tau inversion recovery (STIR)/fat suppressed sequences helpful, 38% found the T2 and contrast sequences helpful, and 26% found the T1 sequence helpful. The top 3 MRI findings thought indicative of active diseases were bone edema (43%), periosteal reaction (37%), and soft tissue inflammation (28%). The top 3 MRI findings most concerning for poor prognosis were vertebral compression (30%), fracture (19%), and physeal irregularity (13%).Among responders who used MRI for disease monitoring (Physicians defined features that indicate active disease as new lesions identified from imaging (92%), elevated ESR and/or CRP (91%), pain localized to known sites (86%), focal bone swelling and/or warmth (83%), focal tenderness at known sites without allodynia (82%), active arthritis (53%), fever (45%), and functional limitation of joints/limbs (24%).Features of inactive disease were defined by physicians as resolution of constitutional symptoms (91%), absent focal tenderness and/or warmth and/or swelling of known CNO lesions (85%), normal erythrocyte sedimentation rate (ESR), c-reactive protein (CRP) (83%), complete resolution of pain at known CNO lesions (71%), resolution of synovitis if present with active disease (71%), absence of inflammation confirmed by imaging (57%), no new CNO lesions confirmed by whole-body MRI (49%), and normal function of the affected sites (37%).Medications that were often or always used by a high proportion of responders included nonsteroidal anti-inflammatory drugs (NSAIDs) . Medications often or always used by a lesser proportion of responders include methotrexate (34%), adalimumab or infliximab (26%), etanercept (17%), glucocorticoids (13%), bisphosphonate (13%), sulfasalazine (4%), other DMARDs (2%), and azithromycin (<1%). These proportions were similar across categories of years of experience, number of patients diagnosed, or number of patients treated (data not shown).\u03b1 inhibitors (65%), and bisphosphonates (46%) were the next most commonly used treatments routinely prescribed an NSAID as initial therapy. For patients who failed NSAID treatment, methotrexate (67%), TNF-eatments . The topeatments . After pSeventy-eight percent of responders have used glucocorticoids to treat CNO. Physicians tended to treat with steroids for short durations: 1\u20133 weeks and 4\u20136 weeks . The most common dosage ranges included 0.25\u20130.5\u2009mg/kg/day , 1\u2009mg/kg/day , or 1-2\u2009mg/kg/day . The maximum daily dose ranged between 10 and 80\u2009mg with a median of 60\u2009mg.p < 0.01) and 1-month longer trial periods for DMARD, biologic, and bisphosphonate (p < 0.01) compared to NSAIDs.The minimum trial period of treatment needed before declaring failure varied across medication classes, with physicians reporting consideration of treatment failure after 2-3 months of use of NSAIDs (63%), nonbiologic DMARDs (77%), or biologic DMARDs (78%) when active disease persisted. However, as early as 1 month of use of glucocorticoids (64%) and as late as 3\u20136 months of use of bisphosphonates (61%) were regarded as treatment failures. Pairwise comparisons between NSAIDs and other drug classes found that, on average, physicians reported 1-month shorter trial periods for glucocorticoid . The presence of a spinal lesion increased the use of bisphosphonate treatment (26% for case\u2009\u20092 versus 4\u20136% for cases\u2009\u20091 and 3). Pamidronate was the most commonly used bisphosphonate followed by zoledronic acid in case\u2009\u20092. The most commonly used dosing of pamidronate was 1\u2009mg/kg/dose monthly or daily for 3 days and repeat every 3 months (maximum dose 60\u2009mg). The first-time dose was often reduced to 0.5\u2009mg/kg in two daily doses for better tolerance. The dosing of zoledronic acid was reported as 0.0125\u2009mg/kg/dose (maximum dose of 4\u2009mg) and repeated every 6 months.Three hypothetical cases were presented. Case\u2009\u20091 had a unifocal lesion in an upper extremity with mildly increased ESR and CRP. Case\u2009\u20092 had vertebral lesions and case\u2009\u20093 had multiple pelvic lesions. The top three leading choices of treatment for all cases were NSAIDs, followed by DMARDs and TNF-Physicians reported continuing treatment for cases\u2009\u20091\u20133 for an additional 4\u20136 months or 7\u201312 months after these patients achieved inactive disease.Fifty-eight percent of responders answered \u201cyes\u201d to being interested in participating in a comparative effectiveness study of different treatments for CNO. Among all responders, the preferred treatment choices for future comparative effectiveness studies in CNO were NSAIDs alone (89%), biologics only (79%), nonbiologic DMARDs only (66%), bisphosphonates (60%), biologics with nonbiologic DMARDs (50%), biologics with bisphosphonates (32%), and bisphosphonates with DMARD (26%) .To our knowledge, this is the first study to assess the practice patterns of pediatric rheumatologists in diagnosing and managing CNO. Using a CARRA-wide survey, we captured the important aspects of disease management including clinical features to obtain bone biopsy, selection of imaging modality, frequency of imaging to monitor disease activity, treatment choices, and treatment duration.Those physicians with more clinical experience in diagnosing and managing CNO were more confident in their abilities caring for these patients than those with less clinical experience caring for patients with CNO. However, the years of practice did not affect the confidence level.Bone biopsy is frequently performed to exclude infection and malignancy but was not considered essential to confirm a diagnosis of CNO by at least 40% of physicians. Interestingly, these physicians used similar criteria to decide which cases to biopsy regardless of reported frequency of ordering bone biopsy.Constitutional changes, unifocal bone lesion, and nocturnal bone pain raise concerns of infection and/or malignancy. Thus, children with these features are often referred for bone biopsy. Although these symptoms are not specific and may be present in some children with CNO, most physicians request a bone biopsy to exclude other causes. Conversely, when multifocal bone lesions exist, typical sites are involved, or associated conditions such as psoriasis, IBD, and ERA are present, physicians may deem bone biopsy unnecessary. Whether ERA is associated with CNO remains controversial. In a pediatric CNO cohort, inflammatory arthritis occurred in up to 80% of children initially or during the course of the disease and five of 30 patients (17%) satisfied the European Spondyloarthropathy Study group criteria for spondyloarthropathy . Our queJansson et al. have developed a scoring system in an attempt to determine the threshold of obtaining a bone biopsy in a child with suspected osteomyelitis . CNO wasIn suspected cases, whole-body MRI has the highest sensitivity to detect active bone lesions , 25. HowOnly a limited number of participants used MRI to monitor disease activity. Characteristics such as bone edema, periosteal reaction, and soft tissue inflammation were identified as indicative of active disease which have been shown to be reversible after treatment in various studies , 15. ChaTreatment options vary for individual patients. Most physicians prescribed NSAIDs as first-line treatment which is in line with published studies , 7, 9. HIn children with CNO who failed NSAIDs, there has not been a consensus treatment plan among pediatric rheumatologists. Our CARRA survey results suggested that nonbiologic DMARDs, TNF inhibitors, and bisphosphonates were most commonly used by the treating physicians and need further study which could be done using comparative effectiveness research.Vertebral lesions were associated with higher risk of compression fracture. In this setting, bisphosphonates have been reported to be effective in reducing the pain and inflammation in bone \u201333. MostThe definition of active and inactive diseases has not been clearly described due to the lack of specific disease markers. Beck et al. developeOur study has several limitations. Firstly, the response rate from pediatric rheumatologists was low; hence there could have been survey-sampling bias. Secondly, there may be deviations between answers to the survey and actual practice. Thirdly, the definition of CNO may vary among physicians.Despite the limitations of our study, we were able to identify key issues such as diagnostic approaches, disease monitoring, and treatment selections after failing NSAIDs as well as definition of disease activity. Results from this study will guide further discussion within a focused group to develop consensus treatment plans for children with CNO.Our results showed that majority of pediatric rheumatologists manage fewer than five patients with CNO per year. Similar to the prevalence of other pediatric rheumatic diseases such as systemic lupus erythematosus and juvenile dermatomyositis, CNO is uncommon and will need a collaborative effort across centers for future prospective clinical studies.The diagnostic approach and disease activity monitoring for CNO varied among physicians. NSAIDs remained the first-line treatment for CNO. Methotrexate, TNF inhibitors, and bisphosphonates were most commonly used after NSAIDs failed. These findings provided important background to move forward with development of consensus treatment plans for CNO.An online survey through Redcaps were used to collect information from CARRA members regarding their practicing patterns of diagnosing and managing children with CNO."} +{"text": "Repetitive Transcranial Magnetic Stimulation (rTMS) commonly is used for the treatment of Major Depressive Disorder (MDD) after patients have failed to benefit from trials of multiple antidepressant medications. No analysis to date has examined the cost-effectiveness of rTMS used earlier in the course of treatment and over a patients\u2019 lifetime.We used lifetime Markov simulation modeling to compare the direct costs and quality adjusted life years (QALYs) of rTMS and medication therapy in patients with newly diagnosed MDD (ages 20\u201359) who had failed to benefit from one pharmacotherapy trial. Patients\u2019 life expectancies, rates of response and remission, and quality of life outcomes were derived from the literature, and treatment costs were based upon published Medicare reimbursement data. Baseline costs, aggregate per year quality of life assessments (QALYs), Monte Carlo simulation, tornado analysis, assessment of dominance, and one way sensitivity analysis were also performed. The discount rate applied was 3%.Lifetime direct treatment costs, and QALYs identified rTMS as the dominant therapy compared to antidepressant medications in all age ranges, with costs/improved QALYs ranging from $2,952/0.32 (older patients) to $11,140/0.43 (younger patients). One-way sensitivity analysis demonstrated that the model was most sensitive to the input variables of cost per rTMS session, monthly prescription drug cost, and the number of rTMS sessions per year.rTMS was identified as the dominant therapy compared to antidepressant medication trials over the life of the patient across the lifespan of adults with MDD, given current costs of treatment. These models support the use of rTMS after a single failed antidepressant medication trial versus further attempts at medication treatment in adults with MDD. Interpreted differently, under reasonable circumstances (e.g. every day practice), rTMS is the less costly treatment alternative providing greater overall treatment efficacy over the life of the patient.A separate (but related) question concerns the cost-effectiveness of rTMS treatment. Using the mid 50s age cohort as the example, the model evaluates how patients \u201cfared\u201d (cost of treatment and QALYs for each stage) in \u201ccycling\u201d through their therapeutic regimens, depending upon the treatment state they are in . rTMS and antidepressant drug therapy outcomes appear in These analyses demonstrate that in patients newly diagnosed with MDD, rTMS can be a more cost-savings (having a higher NMB) /clinically effective therapy than further medication trials after a single failed antidepressant drug treatment\u2014when considered over the entire life of the patient. In other words, rTMS had lower costs and overall higher QALYs over a patient\u2019s lifetime when compared with multiple subsequent attempts of pharmacologic therapy after a single drug failure were Nguyen et al ) and SimThe models presented herein also extend prior work by examining the lifetime effects of treating different aged cohorts of patients on either therapy, in contrast to earlier studies examined non-age specific patients over a tThe sensitivity analysis , rTMS becomes a more costly therapy (having a lower NMB) than medication, with the ICER varying from $29,895 (mid 20\u2019s) to $45,747 (mid 50\u2019s) . TherefoAt the very low end of the range of pharmacotherapy costs per month (around $100/month for generic medications), again rTMS would become the costlier therapy with the ICERs varying from $47,193 (mid 20\u2019s) to $56,875 (mid 50\u2019s) . In thisThe use of rTMS in pharmacoresistant patients (up to four failed pharmacologic treatment regimens) and the clinical efficacy demonstrated in prior analyses may not be reflective of the true clinical efficacy of the treatment, as patients with high levels of medication resistance fare worse in their remission/response rates and become more treatment resistant with each succeeding unsuccessful antidepressant trial ,43,44,45Most prior evaluations of rTMS in high quality studies have examined its efficacy after multiple (\u22652) pharmacologic therapies ,29; althThe conclusions drawn from these Markov models should be interpreted in the context of certain limitations. First, estimates of costs, probabilities, and QALYs were extrapolated over the course of the lifetime of a patient based upon shorter term data. While these extrapolations are similar to those in other published studies , they maSecond, the costs used in the model were Medicare national average reimbursement rates for procedures/services. While for hospital visits, Medicare on average pays approximately 93% of the costs incurred , the relThese models assume that patients using antidepressant medications were adherent to their treatment regimens. A lack of adherence would affect costs and possibly decrease quality of life outcomes of treatment.Lastly, based on the data used, the results would apply to the United States MDD mainly white population. Thus the findings are limited to this group.Markov modeling comparing rTMS and medication treatment outcomes indicates that given current practice standards and costs of MDD treatment, rTMS can be a dominant therapy as compared with antidepressant medication treatment. These models indicate that introduction of rTMS treatment after a single failed antidepressant trials would incur greater cost savings and better outcomes than the current practice of continued successive medication trials. Even under less favorable assumptions, rTMS would be a cost-effective alternative based on ICER threshold guidelines. Thus rTMS should be considered by payers for coverage as an MDD treatment earlier in the course of treatment of adults with MDD.S1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file.S3 Fig(PDF)Click here for additional data file.S4 Fig(PDF)Click here for additional data file.S5 Fig(PDF)Click here for additional data file.S6 Fig(PDF)Click here for additional data file.S7 Fig(PDF)Click here for additional data file.S8 Fig(PDF)Click here for additional data file.S9 Fig(PDF)Click here for additional data file.S10 Fig(PDF)Click here for additional data file.S11 Fig(PDF)Click here for additional data file.S12 Fig(PDF)Click here for additional data file.S13 Fig(PDF)Click here for additional data file.S14 Fig(PDF)Click here for additional data file.S15 Fig(PDF)Click here for additional data file.S16 Fig(PDF)Click here for additional data file.S17 Fig(PDF)Click here for additional data file.S18 Fig(PDF)Click here for additional data file.S19 Fig(PDF)Click here for additional data file.S20 Fig(PDF)Click here for additional data file.S21 Fig(PDF)Click here for additional data file.S1 Appendix(DOCX)Click here for additional data file.S2 Appendix(DOCX)Click here for additional data file.S3 Appendix(DOCX)Click here for additional data file.S4 Appendix(DOCX)Click here for additional data file.S5 Appendix(DOCX)Click here for additional data file.S6 Appendix(DOCX)Click here for additional data file."} +{"text": "Depending on the nature of their items factor analyses of different scales impose different structures on the underlying psychopathological dimensions, so a broader range of scale items should be more revealing. Few studies repeat analyses over successive interviews to investigate whether psychopathology has a consistent structure or evolves, especially after first presentations when the illness is most plastic and cohorts are unselected by chronicity.A cohort was recruited from consecutive presentations aged 16\u201335 to NHS Early Intervention in Psychosis services from 14 catchments over 5 years during the National EDEN project. All met DSM IV-R criteria for schizophrenia spectrum psychoses, brief or substance induced psychoses, mania or severe depression with psychosis. At recruitment, after 6 and 12 months each was assessed with Positive and Negative Symptom Scale (PANSS), Calgary Depression Scale (CDS), Young\u2019s Mania Rating Scale (MRS) and Birchwood\u2019s Insight Scale (IS). At each point principal axis factoring with oblique (Promax) rotation included all scale items simultaneously, apart from using total scores for IS. Items below communality thresholds were excluded and the analyses repeated until stable solutions were achieved with fit metrics meeting conventional thresholds. Factor solutions were selected using breaks in the scree plot and eigenvalues>1.0.1003 met diagnostic criteria and 948 provided data. Each time point produced 6 factors featuring consistent items: psychosis ; excitement/mania/disorganisation ; hostility/suspiciousness ; depression/anxiety ; negative symptoms , and poor insight . Depression explained 29\u201332% of variance at different stages, Psychosis 28\u201329%, Negative 25\u201326%, Excitement 19\u201324%, Hostility 16\u201323% and Poor Insight 16\u201323%.The cohort, recruited from consecutive presentations, included a full range of psychoses in sufficient numbers to factor analyse the scales\u2019 51 parameters. There was evidence for 6 factors slightly different from the traditional 3 SAPS/SANS or 5 PANSS factors derived using chronically unwell samples with non-affective psychosis. There was more consistency than in previous first episode follow-up studies and affective and insight dimensions were more clearly defined."} +{"text": "In a proportion of stroke patients with acute large vessel occlusion permanent stent implantation is mandatory to achieve successful recanalization. The optimum platelet inhibition strategy after such emergency stenting is unknown. We therefore analyzed the outcome of early glycoprotein (gp) IIb/IIIa inhibitor treatment after emergency stenting in acute stroke.Sixty patients with emergency stenting were identified in our stroke unit registry from 12/2010-06/2014 and analyzed retrospectively. All patients were bridged intravenously with the gpIIb/IIIa antagonist tirofiban immediately after the acute procedure until switching to oral aspirin and clopidogrel was performed. For comparison we studied 135 patients with M1 occlusion undergoing thrombectomy without stent implantation or tirofiban treatment in a propensity score-adjusted analysis.In the acute stenting group receiving tirofiban complications with 6 deaths during the hospital stay (10%), 2 reinfarctions (3%), 12 intracerebral hemorrhages and 5 symptomatic ICH (8%) occurred. Thirty-seven patients (62%) reached a moderate outcome of mRS 0\u20133 after 90 days. In the thrombectomy group without tirofiban administration the rate of deaths within hospital stay, the rate of ICH and outcome at day 90 were not different.In our retrospective study acute stenting with subsequent gpIIb/IIIa inhibition was not associated with an increased risk of ICH or in-hospital death. In stroke patients with acute large vessel occlusion and stenoocclusive atherosclerotic lesion stent implantation in addition to clot retrieval may be necessary to access downstream embolic occlusions or ensure lasting recanalization. The optimum platelet inhibition strategy in such situations is currently unclear. Glycoprotein (gp) IIb/IIIa inhibitors such as tirofiban can be administered intravenously, exhibit fast onset of action, and the effect subsides within a few hours after discontinuing infusion. Despite reports of an increased risk of secondary intracerebral hemorrhage (ICH) after ischemic stroke \u20133, tirofAs approved by the local ethics committee [Ethikkomission der Medizinischen Fakult\u00e4t der Heinrich-Heine-Universit\u00e4t D\u00fcsseldorf (#4743R)], routine medical care data of all patients treated for ischemic stroke in the Stroke unit of the Department of Neurology, Heinrich-Heine-University, Duesseldorf from 12/2010\u201306/2014, were collected in an anonymized and pseudonymized manner (n = 2600) and analyzed retrospectively. For observational retrospective analysis a separate written informed consent was not required by the local ethics committee.We identified 60 patients with acute ischemic stroke in the anterior circulation, who received acute stenting of extra- and/or intracranial arteries in addition to endovascular thrombectomy in the same intervention with or without preceding i.v. thrombolysis. All these patients, except for one patient with early ICA stent occlusion during intervention, were treated with the gpIIb/IIIa antagonist tirofiban (1.250 mg bolus during intervention followed by a continuous infusion of 0.1\u03bcg/kg body weight/minute) from time of acute stenting until a switch to aspirin and clopidogrel was performed, mostly within 12\u201324 hours, with 12 hours overlap. For comparison we analyzed 135 patients with ischemic stroke who received endovascular thrombectomy of the middle cerebral artery (M1 segment) occlusion without stent implantation or tirofiban treatment.Imaging was performed with a 3-T or 1.5-T MR scanner or contrast enhanced CT . Alberta stroke program early CT score (ASPECTS) were obtClinical outcome was assessed by trained physicians employing modified Ranking Scale (mRS) at hospital discharge , and aftU test , the MCA (n = 8), the extracranial and intracranial ICA (n = 1) as well as the extracranial ICA and MCA (n = 4). Twelve (20%) patients had an isolated occlusion of the extracranial internal carotid artery with patent M1 or carotid T, 22 (37%) had tandem occlusions including ICA and M1 or carotid T, while 26 (43%) had intracranial occlusions.For comparison, we identified 135 patients who received endovascular thrombectomy of the M1 without stent implantation or tirofiban medication. Pre- and posttreatment ASPECTS was significantly higher in the stenting group. Recanalization with TICI Score \u2265 2b was achieved significantly more frequent in the stenting group.To account for dysbalance in these and other baseline parameters see , outcomeOptimum platelet inhibition strategies following emergency stent implantation in acute ischemic stroke are controversial. Preclinical experimental data in mice showed aDue to a favorable safety profile in the previous SATIS Trial the gpIILimitations of our study are the retrospective design, small sample size, and lack of randomization. Therefore, controlled trials are needed to establish platelet inhibition strategies in acute stroke patients treated by emergency stent implantation."} +{"text": "Papaya ringspot virus (PRSV) isolate from South Korea (SK) infecting squash (Cucurbita pepo) was obtained using paired-end RNA sequencing. A BLASTn search of the PRSV SK isolate full-genome sequence showed nucleotide sequence identity ranging from 81% to 83% with previously reported PRSV isolates (GenBank accession numbers KX655874 and EF017707).The complete genome sequence of a Papaya ringspot virus (PRSV) is a member of the genus Potyvirus (family Potyviridae). It has a single-stranded positive genome of approximately 10,000 nucleotides (nt) that encodes a large polyprotein , and cDNA was synthesized using the N25 primer. cDNA was amplified by PCR using SuPrime Script PCR premix . To determine the 5\u2032 and 3\u2032 terminal sequences, rapid amplification of cDNA ends (RACE) was performed with a 5\u2032/3\u2032 RACE kit according to the manufacturer\u2019s instructions. Amplified DNA was purified and cloned into an RBC T&A cloning vector and sequenced . Three clones were sequenced to determine the complete nucleotide sequence.In May 2014, a The PRSV SK isolate consists of 10,324 nt encoding 3,343\u00a0amino acids, including an 86-nt 5\u2032 untranslated region (UTR) and a 206-nt 3\u2032 UTR. The PRSV SK isolate shows the highest nucleotide sequence identity with the full genome of the PRSV TM50 isolate (GenBank accession number KX655874) (83%) and the PRSV Andong coat protein (KT884449) (97%). The PRSV SK isolate is therefore highly divergent compared with other PRSV isolates.Cucurbita pepo has been deposited in GenBank under the accession number KY996464.The genome sequence of the PRSV SK isolate infecting"} +{"text": "Network analysis identified TNF (tumor necrosis factor), estrogen, and TP53 (tumor protein 53) as the top of 671 upstream regulators (p < 0.001), whereas the SOX2 (SRY [sex determining region Y]-box 2) and OCT4 (octamer-binding transcription factor 4) complex was the top master regulator out of 773 master regulators associated with fertility (p < 0.001). Identification of QTL and genes in pathways that improve early pregnancy success provides critical information for genomic selection to increase fertility in cattle. The identified genes and regulators also provide insight into the complex biological mechanisms underlying pregnancy establishment in cattle.Infertility and subfertility negatively impact the economics and reproductive performance of cattle. Of note, significant pregnancy loss occurs in cattle during the first month of pregnancy, yet little is known about the genetic loci influencing pregnancy success and loss in cattle. To identify quantitative trait loci (QTL) with large effects associated with early pregnancy loss, Angus crossbred heifers were classified based on day 28 pregnancy outcomes to serial embryo transfer. A genome wide association analysis (GWAA) was conducted comparing 30 high fertility heifers with 100% success in establishing pregnancy to 55 subfertile heifers with 25% or less success. A gene set enrichment analysis SNP (GSEA-SNP) was performed to identify gene sets and leading edge genes influencing pregnancy loss. The GWAA identified 22 QTL (p < 1 x 10 Fo. Fo67]. opoiesis . IL6 haselopment .CTNNB1, GJA1, BMP4, WNT2, WNT16, and GATA3) were present in three or more gene sets (CTNNB1), also known as beta-catenin. CTNNB1 is involved in the regulation of GJA1, BMP4, and GATA3 whereas CTNNB1 has a feedback loop with WNT2 and WNT16 in the WNT (wingless) signaling pathway. CTNNB1 plays a crucial role in adherens junctions in cell-cell adhesions and is the central effector in the canonical WNT signaling pathway [WNT signaling is likely an important functional mediator of early pregnancy events in cattle, sheep, mice and humans [Out of 253 unique leading edge genes in nine gene sets associated with heifer fertility, six genes that are directed by the master regulator SOX2-OCT4 (sex determining region Y box 2 \u2013octamer-binding transcription factor 4) complex .\u221217) upstream regulator, TNF, directly regulates 66 genes associated with heifer fertility. TNF produces a cytokine protein with systemic and local functions that are elicited in response to infection and inflammation. In the uterus, the primary source of TNF is uterine macrophages and natural killer cells. TNF also stimulates nuclear factor kappa B (NFKB) to trigger inflammation [Rac1 is an upstream regulator of P38 mitogen-activated protein kinases (P38 MAPK) [Rac1 and MAP kinases are leading edge genes in this data set (The most significant (p = 3.3 \u00d7 10ammation . Rac1 is38 MAPK) and bothdata set . Togethe\u221216) that controls 64 genes associated with heifer fertility. Optimal circulating levels of maternal estrogen and progesterone is critical to maintain implantation in humans and mice, as an imbalance of estrogen can render the uterus unreceptive to implantation [WNT signaling pathway [WNT signaling genes along with beta-catenin are leading edge genes in this data set (Beta-estradiol is an upstream regulator (p = 1.9 \u00d7 10antation . Estroge pathway . Many WNdata set . Previou\u221215). TP53 encodes an anti-apoptotic protein known as a tumor suppressor that regulates the cell cycle and DNA repair. However, TP53 also has a role in female fertility and implantation. Polymorphisms in intron 3 and exon 4 of TP53 were associated with endometriosis and post-in vitro fertilization implantation failure in women due to alterations in leukemia inhibitory factor (LIF) expression [LIF has a diverse role in embryo implantation including transformation of the endometrium to a receptive stage, stromal decidualization, uterine leukocyte migration and regulation of prostaglandin synthesis [A third upstream regulator of fertility was TP53 which controls 60 genes associated with heifer fertility . Transcriptional regulators SOX2 and OCT4 are two main components of the embryonic stem cell pluripotency circuit. These factors may also be important for endometrial regeneration, repair and remodeling during pregnancy in cattle, mouse and human [OCT4 and SOX2 was in the stromal cells in the ipsilateral horn of the uterus [The master regulator identified was the SOX2-OCT4 complex, which regulates 18 upstream regulators and 4 GWAA positional candidates nd human . Regenernd human . OCT4 ane uterus . Highly in vitro produced embryos transferred into recipient heifers. The analysis of SNP data through GWAA, GSEA-SNP and IPA revealed QTL and candidate genes that regulate establishment of early pregnancy in cattle. These approaches facilitate our understanding of the genetics and biology of endometrial receptivity in cattle. Further validation of these results in independent populations are needed before the QTLs identified from this study can serve as fertility markers for genomic selection. Validation of these markers in independent populations will also be helpful in the identification of causal mutations, which is desirable because they provide heightened accuracy when using genomic selection, particularly in multiple breeds.This study focused on the QTL and genes associated with the loss of pregnancy in cattle through failed implantation of the embryo by using Many of the gene sets and pathways identified in this study include genes already implicated in fertility in other species. Further investigation is needed to determine the role of the key upstream and master regulators that control many of the positional candidate genes identified in this study as associated with the establishment of pregnancy. The understanding of these genes, pathways and regulators is expected to help reduce early pregnancy loss in cattle resulting in increased reproductive performance.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file."} +{"text": "Diffuse myocardial fibrosis is an important pathophysiological process involved into essential hypertension which may increase myocardium stiffness, cause ventricular remodeling and increase adverse cardiovascular risk eventually. It is strongly evident that optimal antihypertensive therapy will reduce the risk induced by hypertension, however, very few proportion of patients with hypertension can reach the recommended target of blood pressure (BP) control. So far little knowledge has been illuminated for effect of blood pressure control status on the myocardial fibrosis in hypertension. T1 mapping and extracellular volume (ECV) quantified by CMR has been adopted as a good method in examination of diffuse fibrosis. Besides, variation of serum fibrosis biomarkers may also reflect the degree of myocardial fibrosis. Based on the above we hypothesised that there was a correlation between ECV, serum fibrotic biomarkers and long-term BP control status in patients with essential hypertension.T1 mapping by using modified Lock-Locker inversion recovery(MOLLI) sequence on a 3.0 Tesla scanner was performed on 40 hypertension patients (20 patients with documented well BP control and 20 with uncontrolled BP) and 20 healthy controls. Myocardial native, post T1 and average ECV of left ventricle were measured based on pre contrast and post contrast T1 mapping(15 minutes after intravenous injection with gadopentetate dimeglumine (0.15 mmol/kg). Serum PICP and ICTP level were detected with ELISA kits.\u03b2 = 0.37, p = 0.02, PICP/ICTP: \u03b2 = 0.30, p = 0.03) (Table Uncontrolled hypertensive patients had a higher interventricular septum thickness(IVS) or left ventricular mass index(LVMI) than well controlled hypertensive patients, whereas there was no difference in left ventricular functional parameters between two groups. Hypertensive patients had higher mean ECV of left ventricular myocardium than normal controls, furthermore, mean ECV of LV in hypertensive patients with uncontrolled BP was higher than in patients with ECV well- controlled,whereas the differences of native or post-contrast T1 time of LV myocardium between the two groups was not statistically significant (Table ECV of LV myocardium was higher in hypertensive patients which was associated with BP control status. ECV could be associated with the LV hypertrophy in patients with uncontrolled hypertension and could also reflect the changes of myocardial fibrosis in hypertension."} +{"text": "Citrus tristeza virus (CTV) from Uruguay, sequenced by using Illumina and Sanger sequencing technology. This CTV DSST-17 genome clustered within genotype resistance breaking (RB) and presents two recombination events.We report here the complete genome sequence of a Citrus tristeza virus (CTV) is one of the most destructive pathogens that affects citrus trees around the world and has been responsible for the loss of over 100 million trees in the past 70\u2009years that encode at least 19 proteins , 5. The ifoliata . Last yeifoliata .de novo assembly a long contig of 19,269\u2009nt. The complete genome obtained was compared with all available full genomes of the GenBank database using MEGA 6 and submitted to RNA library preparation with a NEBNext Ultra II RNA library prep kit (Illumina). The library was sequenced using the NextSeq 500 system platform (Illumina). Reads were trimmed and assembled with CLC Genomics Workbench version 11. After trimming, reads with an average length of 150 nucleotides (nt) were used to generate through g MEGA 6 . Phylogeg MEGA 6 . The anaMH186146.The genomic sequence for isolate CTV DSST-17 was deposited in GenBank under accession number"} +{"text": "Our findings elucidate the novel downstream pathways of CSPGs and suggest potential synergy of blocking their two PTP receptors.Receptor protein tyrosine phosphatase \u03c3 (PTP\u03c3) and its subfamily member LAR act as transmembrane receptors that mediate growth inhibition of chondroitin sulfate proteoglycans (CSPGs). Inhibition of either receptor increases axon growth into and beyond scar tissues after CNS injury. However, it is unclear why neurons express two similar CSPG receptors, nor whether they use the same or different intracellular pathways. We have now studied the signaling pathways of these two receptors using N2A cells and primary neurons derived from knockout mice. We demonstrate that both receptors share certain signaling pathways , but also use distinct signals to mediate CSPG actions. Activation of PTP\u03c3 by CSPGs selectively inactivated CRMP2, APC, S6 kinase and CREB. By contrast LAR activation inactivated PKC\u03b6, cofilin and LKB1. For the first time, we propose a model of the signaling pathways downstream of these two CSPG receptors. We also demonstrate that deleting both receptors exhibits additive enhancement of axon growth in adult neuronal cultures After CNS injuries, a family of extracellular matrix (ECM) molecules, the chondroitin sulfate proteoglycans (CSPGs), are highly upregulated by reactive scars and potently inhibit axon growth into and beyond the lesion area1246in vitro910121315161718The presence of sulfated GAG chains is particularly important because removing GAGs or preventing GAG sulfation neutralizes most of the suppression of axon growth by CSPGs in vitro.Deficiency of PTP\u03c3 or LAR in adult mice increased regrowth of various projection tracts after spinal cord injury (SCI), including sensory, corticospinal and serotonergic axons, into the caudal spinal cord151716This study supports a novel model of signaling pathways downstream of the two CSPG receptors and potential synergistic actions of blocking both simultaneously.RhoA is a prominent intracellular signal that regulates cytoskeletal dynamics, transcription and cell growth, and is thought to mediate functions of multiple axon growth inhibitors, including myelin- associated inhibitors21232628Treatment with CSPGs for 20\u2009min significantly increased active RhoA levels in N2A cells transfected with either PTP\u03c3 or LAR, compared with those in control cells . BecauseTo validate the findings from N2A cells in primary neuronal cultures, we evaluated whether PTP\u03c3 or LAR deletion alters activities of intracellular signals, including active RhoA, in cultured cerebellar granule neurons (CGNs) following incubation in CSPGs. We could easily collect enough samples for multiple Western blot analyses using CGN cultures derived from postnatal 7\u201310\u2009day mice. Following 24\u2009hours of growth, CGNs were treated with CSPGs (1.5\u2009\u03bcg/ml) for 20\u2009minutes and the levels of active RhoA in the supernatants were detected by precipitation with rhotekin binding domain beads and then by Western blotting. Application of CSPGs at 1.5\u2009\u03bcg/ml significantly enhanced levels of active RhoA in CGNs derived from PTP\u03c3+/+ mice . HoweverAs a downstream mediator of RhoA action, Rock can phosphorylate and activate LIM kinase (LIM-K), which in turn phosphorylates and inactivates cofilin3032The intracellular PI3K/Akt pathway is essential for regulating neuronal growth during development and contributes to suppression of axon growth by CSPGs16212236Deletion of PTEN, a negative regulator of PI3K/Akt signaling, stimulated neuronal growth and survival following axotomy by activating mTOR38CRMP2 is critical for regulating axon formation and outgrowth during development by binding tubulin heterodimers and promoting microtubule assembly4243454647The tumor suppressor APC accumulates at the tip of the growth cone during axon elongation and its phosphorylation by GSK-3\u03b2 suppresses axon growth by reducing its ability to bind microtubules1621Extracellular regulated kinases (Erks) are expressed in multiple cell types including neurons, and are essential for regulating protein synthesis/degradation and maintaining levels of proteins required for growth in response to many extracellular axon growth and guidance cues, including neurotrophins. Although it has not been determined whether Erk signaling mediates CSPG function, nerve growth factor (NGF) promoted growth of conditioning DRG neurons cultured on CSPGsThe 90\u2009kDa ribosomal S6 kinases (p90RSKs), also known as MAPK-activated protein kinase 1, are Ser/Thr kinases characterized by two non-identical functional kinase domains and a carboxy-terminal docking site for Erks51Activation of Erk can enhance CREB phosphorylation at serine 133, thus increasing its activity as an upstream signal53The serine/threonine kinase 11 (known as LKB1) and its downstream signaling target SAD/MARK regulate neuronal polarization and axon elongation during development5758As a member of the atypical protein kinase C subfamily, PKC\u03b6 phosphorylates LKB1 at Ser-428/431 and mediates activation of AMPK in endothelial cells60in vitro by counting the number of neurites that cross a gradient of CSPGs1516in vitro.Our experimental results with either N2A cells or primary neurons support that LAR and PTP\u03c3 share certain signaling pathways, but also employ distinct signals to covey CSPG effects on neurons. It was thus important to determine whether targeting both PTP receptors would have additive actions in overcoming CSPG-mediated inhibition of axon growth. CSPG-spot assay is a reliable method for measuring axon growth To compare downstream signals of the two RPTPs, we conducted parallel experiments, measuring alterations of multiple signaling proteins in the lysates of PTP\u03c3- or LAR-overexpressing N2A cells. These neuroblastoma-derived cultures possess the functional properties of neurons27656621223435Based on the results of the present study and previous reports, we propose a simplified model of convergent and divergent pathways downstream of the two PTP CSPG receptors . The int323472PTP\u03c3 and LAR do not employ identical signaling to mediate CSPG actions, although both use the Akt/GSK-3\u03b2 signaling pathway . ActivatAs a downstream signal for many neurotrophic factors747659Moreover, activation of PTP\u03c3, but not LAR, by CSPGs inactivated CREB by reducing its phosphorylation, suggesting that CSPG-PTP\u03c3 interactions inactivate Erk and subsequently reduce the activity of CREB, an important transcription factor for controlling neuronal growth5354As a downstream signal of Erk, LKB1 appears to be critical for regulating axon elongation during developmentWhy PTP\u03c3 and LAR employ both convergent and divergent downstream signaling pathways remains unclear. It may relate to diversity in the ligands and substrates of RPTPs, indirect links along the different intracellular pathways and multiple interactions among the signals. Although CSPGs and Although HSPGs appear to be the substrates of PTP\u03c3 and LAR1516846889091To explain how PTP\u03c3 and LAR differ in their CSPG-signaling functions, we cannot exclude possible subtle roles of other signaling proteins that did not appear significantly altered by CSPG actions in the current experiments. For example, given the trend of their slight alterations , 4E-BP1 94We detected a crucial role of atypical PKC\u03b6, but not the conventional PKC, in regulating CSPG-LAR interactions . In cont17In this study, we examined 15 signaling proteins that potentially act as the downstream pathways mediating the interactions between CSPGs and two PTP receptors and usedIn identifying the signaling pathways of the two CSPG receptors, we primarily employed one-way ANOVA to determine whether CSPG stimulation induces statistically significant differences at several time points. The overall alterations of the signaling proteins in N2A cells and CGN cultures matched well. Because the properties of N2A cells may not be identical to those of primary CGNs, the extents and latencies of the signaling changes might not be the same. For example, p-S6 showed a greater degree of reduction at more delayed time points in CGNs than in PTP\u03c3-transfected N2A cells . HoweverOn the basis of the present results and previous studies, we summarize convergent and divergent pathways that mediate the axon growth-inhibitory effects of CSPG-RPTP interactions in in vitro to a greater degree than deletion of either receptor alone or transcription factors (such as KLFs)389899Transgenic or pharmacological inhibition of either PTP\u03c3 or LAR induces significant regrowth of several projection fiber tracts after SCI, including serotonergic, corticospinal and sensory fibers, and improves functional recovery16181516or alone . MoreoveAll experimental protocols were approved by the Institutional Animal Care & Use Committee at Temple University and the methods were carried out in accordance with the relevant guidelines and regulations.Antibodies against the following proteins were used: mouse mAb phospho-Akt , rabbit mAb phospho-S6 ribosomal protein (Ser235/236), rabbit mAb phospho-Erk1/2 (p44/42), rabbit pAb phospho-CRMP2 (Thr514), rabbit pAb phospho-cofilin (Ser3), rabbit mAb phospho-4E- BP1 , rabbit mAb phospho-CREB , rabbit mAb phospho-PKA C , rabbit pAb phospho-90\u2009kDa ribosomal S6 kinase , rabbit pAb phospho-PKC\u03b6/\u03bb (Thr410/403), rabbit mAb phospho-PKC (pan) , rabbit mAb phospho-LKB1 , rabbit pAb phospho-MAP1B , rabbit pAb phospho-APC , mouse anti-actin clone C4 and mouse mAb against RhoA . The major proteins employed include rhotekin binding domain beads , a mixture of purified CSPGs , laminin (Sigma) and several protease inhibitors . A plasmid with the human LAR sequence was provided by Dr. Morgan Sheng and amplified in our lab16N2A, a mouse neural crest-derived cell line, possesses neuronal properties656667102PTP\u03c3 and LAR knockout mice were provided by Drs. Michel TremblaySupernatant samples from N2A or CGN cultures (in 35\u2009mm dishes) were prepared as above. After quantification of total proteins in lysates, using Bio-Rad DC protein assay reagents, a sample containing the same amount of proteins from each dish was incubated with rhotekin binding domain coupled beads (45\u2009\u03bcg/sample) for 50\u2009min at 4\u2009\u00b0C. GTP-bound RhoA and total RhoA in cell lysates were detected by Western blot using a mouse monoclonal antibody against RhoA\u00ae scanner and Odyssey\u00ae imaging software. For the Western blotting assays, at least three separate experiments were performed and the representative blots are shown in the figures.Expression changes of each studied signaling protein were determined with Western blots in N2A cells and CGNs after CSPG treatment. The supernatants of cell lysates containing the same amount of total proteins were loaded onto Tris-Glycine gels and transferred to nitrocellulose membranes. The membranes were blocked with 5% blotting grade milk (BIO-RAD), blotted with primary antibodies described above, and then incubated with appropriate secondary antibodies conjugated to IRDye 800CW (LI-COR) or IRDye 680RD (LI-COR). To determine the levels of loading control proteins, membranes were probed with anti-total signaling proteins and/or actin using separate membranes with the same amount of total loading proteins. Visualization and quantification were carried out with the LI-COR OdysseyDRGs were harvested from various genotypes of mice . Quantification data for each signaling protein were collected from 3\u20135 separate experiments.SigmaPlot software was used for statistical analysis. Data in graphs are shown as means\u2009\u00b1\u2009SEM. The comparisons between multiple groups were analyzed with a repeated measures ANOVA. The experiments comparing a single determination of means between two independent groups, including the active RhoA and neurite growth assay, were analyzed with Student\u2019s How to cite this article: Ohtake, Y. et al. Two PTP receptors mediate CSPG inhibition by convergent and divergent signaling pathways in neurons. Sci. Rep.6, 37152; doi: 10.1038/srep37152 (2016).Publisher\u2019s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations."} +{"text": "Cyp51\u2212/\u2212) from cholesterol synthesis is characterized by the progressive onset of liver injury with ductular reaction and fibrosis. These changes begin during puberty and are generally more aggravated in the knockout females. However, a subgroup of (pre)pubertal knockout mice (runts) exhibits a pronounced male prevalent liver dysfunction characterized by downregulated amino acid metabolism and elevated Casp12. RORC transcriptional activity is diminished in livers of all runt mice, in correlation with the depletion of potential RORC ligands subsequent to CYP51 disruption. Further evidence for this comes from the global analysis that identified a crucial overlap between hepatic Cyp51\u2212/\u2212 and Rorc\u2212/\u2212 expression profiles. Additionally, the reduction in RORA and RORC transcriptional activity was greater in adult HCyp51\u2212/\u2212 females than males, which correlates well with their downregulated amino and fatty acid metabolism. Overall, we identify a global and sex-dependent transcriptional de-regulation due to the block in cholesterol synthesis during development of the Cyp51 knockout mice and provide in vivo evidence that sterol intermediates downstream of lanosterol may regulate the hepatic RORC activity.Development of mice with hepatocyte knockout of lanosterol 14\u03b1-demethylase (H The liver is to date the best example of a non-reproductive tissue that shows major differences in gene expression between males and females. The majority of liver-related sex differences emerge during pubertal developmentHmgcr\u2212/\u2212 mice survive for up to 5 weeksCyp51\u2212/\u2212 mice, survive for over 19 weeks although with marked changes in liver structure and function111617The development of liver begins at embryonic (E) day 8.5\u20139, with hepatocyte differentiationCyp51\u2212/\u2212 mice and show that normal liver function becomes impaired by the end of puberty, with earlier presentation and enhanced liver damage in females. A subgroup of male prevalent knockout mice exhibits augmented liver dysfunction combined with abnormal hepatic sterol profiles. Lastly, our work highlights the need for unperturbed cholesterol synthesis for normal RORC signalling in vivo.Herein we examine the development of the hepatocyte Cyp51 knockout mice (HCyp51\u2212/\u2212) on the wild type (Cyp51+/+)Cyp51+/\u2212) background were born in accordance with the Mendelian ratio . Runts were exclusive to the HCyp51\u2212/\u2212 genotypes. They died or had to be euthanized at 4\u201310 weeks with a male to female ratio of 2:1 exhibited severe developmental abnormalities and about half of males (4/7) at 6 weeks. Enhanced ductular proliferation, inflammation and mild-to-moderate portal fibrosis were seen in all HCyp51\u2212/\u2212 mice at 9 weeks (Haematoxylin and eosin (HE) and collagen staining (Sirius red) revealed initial signs of ductular reaction with mild inflammation (lymphocytes and granulocytes) surrounding the portal vein in all H 9 weeks and were 9 weeks .Cyp51 excision on liver development, we performed gene expression profiling using Affymetrix microarrays on Cyp51+/+ and HCyp51\u2212/\u2212 mice, aged 3, 6 and 19 weeks, of both sexes, including runts. The number of DE genes between HCyp51\u2212/\u2212 and control mice increased progressively from 3 weeks onward (Cyp51+/+ mice (Cyp51\u2212/\u2212 mice (202/287 pathways) . Similars) et al.. This po250 TFs) . In both250 TFs) . Reduced250 TFs) , suggestCyp51\u2212/\u2212 mice, but remained unchanged in runts. This was true also for esterified and free cholesterol mice and compared the transcriptome data between the two models. We found 50 DE genes separating HRorc\u2212/\u2212 and control mice and ZT19 (night-time RORC zenith). TF enrichment of the transcriptome data expectedly showed increased night-time RORC activity only in the control mice since its (circadian) expression is dampened in HRorc\u2212/\u2212 livers. From 1264 diurnal DE genes in control mice, 791 lost their rhythm due to RORC abolishment in the HRorc\u2212/\u2212 mice and may be considered as potential RORC targets. Moreover, 105 genes from this list were positive for RORC binding based on Chip-seq results by Takeda et al.We next set out to evaluate the contribution of diminished RORC activity on the phenotype of Hrol mice , where 1rol mice and 5 asCyp51 disruption was consistently higher in HCyp51\u2212/\u2212 females compared to the males . The initial response in 6-week HCyp51\u2212/\u2212 females favoured apoptosis and ErbB signalling, pointing to enhanced liver damage and regeneration. At 19 weeks, decreased fatty and amino acid metabolism and enhanced inflammatory and cancer-related pathways were also underlined (Casp12 indicating ER stress and activated unfolded protein response (UPR) represents a hallmark of male runts. TF enrichment also showed elevated activity of NFYs (p\u2009=\u20090.06) and other damage response factors . KEGG pathway enrichment showed better preservation of metabolism and milder inflammation in runt females.The progressive increase in the number of DE genes between males and females is in agderlined . Male seCyp51\u2212/\u2212 females, which correlates well with their reduced amino and fatty acid metabolism. These data further implicate ROR signalling as one of the sex-dependent metabolic regulators.Expression of RORA and RORC is sexually dimorphic with RORC being generally higher in males and RORA in femalesCyp51\u2212/\u2212 mice became visible at 6 weeks of age and TFs (NFY), and decreased peroxisomal metabolism in 19-week HCyp51\u2212/\u2212 and runt mice. The steep rise in DHL might therefore represent an initiator of hepatocellular stress and liver damage . Conversion of DHL to one of the downstream intermediates from the Kandutsch-Russell branch of cholesterol synthesis could also explain the paradoxically increased lathosterol and 7-dehydrocholesterol in runts. Recently, it was also discovered that cholesterol biosynthesis intermediates act as ROR\u03b3t ligands in Th17 cells11Cyp51 overexpression that metabolizes both sterolsAn interesting question is why is there such a large increase in DHL in runts as opposed to the other HCyp51\u2212/\u2212 and runt mice resulted in diminished RORC activity that could contribute to a fall in the overall metabolic capacity and were conducted in agreement with the European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes (ETS 123) as well as with the guidelines for work with laboratory animals by the National Institute of Health. In the case of Rorc mice, experiments were performed in accordance with protocols approved by the Institutional Animal Care and Use Committee of the NYU School of Medicine.Mice experiments were performed in compliance with the current guidelines and regulations. In the case of Cyp51 knockout mice were prepared by crossbreeding Cyp51flox/flox or Cyp51-/flox with Alb-Cre mice to generate hepatocyte specific Cyp51\u2212/\u2212 mice on a wild type and heterozygous background8Cyp51flox/flox (Cyp51+/+) and heterozygous Cyp51-/flox (Cyp51+/\u2212) mice were used as controlsHepatocyte Rorc knockout mice (HRorc\u2212/\u2212) were prepared by crossbreeding Rorcflox/flox and Alb-Cre mice. Hepatocytes were isolated from 12\u201313 week old mice at ZT7 and ZT19 using the two-step collagenase perfusion method.Hepatocyte Formalin-fixed and paraffin-embedded liver sections were cut at 5\u2009\u03bcm and stained with hematoxylin and eosin for general histological evaluation or with Sirius red to assess collagen deposition.Plasma parameters were measured using commercially available kits. Liver sterols were evaluated by coupled gas spectrometry/mass spectrometry as previously describedCyp51+/+ and Cyp51+/+ HCyp51\u2212/\u2212 mice of both sexes, aged 3, 6, 19 weeks and runts (30 samples). Gene expression analysis was done in R using Bioconductor packages limma and PGSEA. Gene sets for enrichment analysis were taken from the KEGG and TRANSFAC databases.Affymetrix GeneChip\u00ae Mouse Gene 2.0 ST arrays were used to conduct gene expression profiling on Libraries were prepared using the Illumina TruSeq Stranded Total RNA library prep, with Ribozero Gold, starting from 500 ng of total RNA and loaded on high output HiSeq 2500 flow cells. RNA alignments were performed using STAR and reads mapping to exons counted using featureCounts. Counts were normalized in R using the DESeq2 package.Detailed protocols can be found in the How to cite this article: Urlep, Z. et al. Disrupting Hepatocyte Cyp51 from Cholesterol Synthesis Leads to Progressive Liver Injury in the Developing Mouse and Decreases RORC Signalling. Sci. Rep.7, 40775; doi: 10.1038/srep40775 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations."} +{"text": "The group II metabotropic glutamate receptor 3 (mGlu3) is an emerging therapeutic target for schizophrenia, as research has demonstrated a link between mutations in the human gene encoding for mGlu3, GRM3, and clinical diagnosis of schizophrenia. Schizophrenia is known to be accompanied by debilitating cognitive impairments that negatively impact the overall quality of life of the patient. While current pharmacological therapeutics mainly target the positive symptoms, cognitive symptoms are often not effectively treated. Our recent discovery that mGlu3 and mGlu5 can act as signaling partners to modulate synaptic plasticity in the prefrontal cortex led us to hypothesize that mGlu3 may subserve similar functions to those of mGlu5 during hippocampal synaptic plasticity and hippocampal-dependent behaviors.We directly tested this hypothesis using acute slice electrophysiology to investigate basal synaptic transmission as well as long-term plasticity in hippocampal slices. To test cognition, the associative fear learning behavioral assay, termed trace-fear conditioning, was used. C57bl/6 mice or CaMKII-cre;mGlu5-/- mice were used in all studies.We report that mGlu2/3 activation enhances hippocampal theta-burst (TBS)-induced LTP but was without effect on group I mGlu agonist-induced LTD The group II mGlu agonist enhancement of TBS-LTP was blocked by antagonists of mGlu3 or mGlu5.We next tested downstream mechanisms of group II mGlu induced LTP by chemically activating LTP with the group II agonist LY379268 in combination with selective antagonists. We verified the LTP was induced by mGlu3 activation but not mGlu2 using selective negative allosteric modulators of each subtype. Furthermore, mGlu5 negative allosteric modulation with MTEP blocked mGlu3-LTP, and conversely the mGlu5 positive allosteric modulator, VU0092273, enhanced mGlu3-LTP. The cannabinoid receptor type 1 antagonist AM251 was also capable of blocking mGlu3-LTP, suggesting cannabinoid signaling mechanistically drives this LTP.Having confirmed a role for mGlu5 in the mGlu3-LTP, we next verified that postsynaptic mGlu5 located on pyramidal neurons was necessary for mGlu3-LTP by utilizing CaMKII-cre;mGlu5-/- mice. It was found that hippocampal slices from these mice showed no enhancement of LTP when LY379268 was bath applied alone or in combination with TBS-stimulation.Behaviorally, we discovered that selective activation of mGlu3 by systemically injecting the group II mGlu agonist in combination with a selective mGlu2 negative allosteric modulator, VU6001966, causes an enhancement in the acquisition of trace-fear conditioning learning. This was also confirmed to be dependent on mGlu5 as both systemic pharmacological inhibition or genetic deletion of mGlu5 abolished this learning enhancement. Further testing of the ability of mGlu3 activation to augment other cognitive tasks is currently underway.These results taken together demonstrate mGlu3 enhances hippocampal LTP and hippocampal-dependent learning through mechanisms that involve both mGlu5 and CB1 receptor activation. This work provides a basic biological mechanism and preclinical therapeutic validation for mGlu3 as a target for neurological disorders in which cognition is disrupted such as schizophrenia."} +{"text": "We conducted an external quality assessment of Zika virus molecular diagnostic tests in Brazil using a new Zika virus standard. Of 15 laboratories, 73% showed limited sensitivity and specificity. Viral load estimates varied significantly. Continuous quality assurance is required for adequate estimates of Zika virus\u2013associated disease and determination of patient care. The catastrophic Zika virus outbreak in the Americas has affected millions of persons. Brazil was the most affected country and reported \u224895% of all cases of suspected Zika virus\u2013associated congenital disease in Europe revealed that 60% of laboratories need to improve molecular Zika virus detection . This finding suggests a potential lack of sensitivity that may be problematic given that viral loads of 103\u2013104 copies/mL are commonly observed in Zika virus\u2013infected patients reported correct results for all samples. Five (33%) reported 1 or 2 false-negative results from samples with low Zika virus concentrations , panel A>3 false results, including at least 2 false-positive detections of Zika virus\u2013negative specimens. No heterologous flavivirus was particularly affected by false-positive detection, suggesting that false-positive results did not result from unspecific binding of assay oligonucleotides (Six (40%) laboratories reported leotides . InsteadEQA performance varied according to the way viral RNA was prepared. The 8 laboratories conducting Zika virus detection using automated platforms performed generally superior compared with the 7 laboratories conducting manual RNA extraction . This fiAs previously reported , panel B10 median deviation between results (p = 0.429 by Wilcoxon signed rank test). This observation suggests usability of the armored RNA for Zika virus quantification in tropical regions. Irrespective of the standard, viral load determinations among laboratories were comparable with 0.12\u20130.88 log10 median deviations of viral load estimates among laboratories for individual Zika virus specimens. However, we also observed drastic deviations of up to 6 orders of magnitude (Quantification of Zika virus loads did not differ significantly between use of the armored RNA and the WHO Zika virus standard, with only 0.76 logagnitude , suggestSome laboratories in Brazil showed suboptimal sensitivity and specificity of Zika virus diagnostic testing. However, these laboratories performed comparably to those in Europe (Independently of the challenges of Zika virus molecular detection, because of taxation and distributor margins, RT-PCR reagents in Latin America are usually 100%\u2013200% more expensive than in affluent countries (Finally, lack of sensitivity directly affects estimates of the absolute risk for Zika virus\u2013induced congenital disease upon maternal infection during pregnancy ("} +{"text": "CRISPR/Cas9 is now regularly used for targeted mutagenesis in a wide variety of systems. Here we report the use of ribozymes for the generation of gRNAs both in vitro and in zebrafish embryos. We show that incorporation of ribozymes increases the types of promoters and number of target sites available for mutagenesis without compromising mutagenesis efficiency. We have tested this by comparing the efficiency of mutagenesis of gRNA constructs with and without ribozymes and also generated a transgenic zebrafish expressing gRNA using a heat shock promoter (RNA polymerase II-dependent promoter) that was able to induce mutagenesis of its target. Our method provides a streamlined approach to test gRNA efficiency as well as increasing the versatility of conditional gene knock out in zebrafish. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology has revolutionized biology by greatly simplifying targeted mutagenesis . The metZebrafish researchers have been quick to adopt CRISPR/Cas9 technology for the generation of germ line mutations in their model system ,2. Typicin-vivo expression of gRNAs. Current methods use the zebrafish RNA polymerase (RNAP) III-dependent U6 promoter, which produces transcripts with well-defined start and end sites, to express gRNA in zebrafish [in-vivo mutagenesis can only be achieved by expressing the Cas9 endonuclease under the control of tissue-specific RNAP II promoters. The use of similar promoters to drive gRNA expression would greatly increase the versatility of CRISPR/Cas9 for conditional gene knock out; however, unlike the bacteriophage and RNAP III promoters currently used to generate gRNAs in vitro and in vivo respectively [A potentially powerful extension of the application of CRISPR/Cas9 technology is the generation of somatic mutations by the ebrafish ,6. The uebrafish . Howeverebrafish . Moreoveectively ,8 the exectively .in-vivo expression of gRNAs using RNAP-II dependent promoters both in yeast and mammalian cell lines [in-vitro and in-vivo for genome modification in zebrafish. Our method streamlines the generation of generation of gRNAs and increases the versatility of CRISPR/Cas9 mediated conditional mutagenesis in the zebrafish.Ribozymes are RNA molecules with catalytic activity and catalysis occurs utilizing sequence specific interactions within the RNA molecule . In partll lines ,14. In tTg(smyhc1:GFP)i104 transgenic line has been previously described [Fish were maintained in the IMCB zebrafish facility and embryos were obtained through natural crosses and staged according to Kimmel et al. . All expescribed .Tg(smhyc1:GFP)i104 was made using PI-SceI therefore Tol2 transposase should not remobilize the transgene. Transgenic zebrafish were generated by injecting embryos with 30 fg Tol2 and 30 fg Plasmid DNA.For CRISPR mutagenesis, embryos were injected with 200 fg Cas9 mRNA and 50 fg gRNA or 300 fg mRNA containing mCherry and gRNA (this reflects the amount of RNA injected in each embryo). To test the efficiency of DNA constructs expressing gRNA, 30 fg of plasmid and 20 fg Tol2 RNA was injected into embryos. Whole mount immunohistochemistry was performed as previously described . Primarysmo gRNA was made using the hsp70l promoter and the Tol2 destination vector containing cmlc2:GFP marker from the Tol2 kit. Plasmids will be made available in Addgene.PCR amplification for cloning was done using PrimeSTAR Max . gBlocks containing the HH-gRNA and HH-gRNA-HDV constructs were synthesized by IDT (Integrated DNA technologies) and final constructs were assembled using Gibson assembly . Middle entry vector containing Cas9 was cloned from pCS2-nCas9n . Larger mRNA was transcribed using Sp6 mMessage mMachine kit . gRNA was synthesized using the MEGAshortscript T7 kit . RNA was purified by lithium chloride and ammonium acetate for the mMessage mMachine kit and MEGAshortscript kit respectively. Further purification of RNA containing ribozyme is not necessary. Ribozyme cleavage of RNA was determined by denaturing 10% PAGE. Briefly, samples were resuspended in 50% formamide, 10mM EDTA and heated to 70\u00b0C for 5 min and chilled on ice. Denaturing PAGE gels contain 1xTBE and 7 M urea. PAGE Gel was pre-run at 20 W for 15 min before loading samples. Electrophoresis was carried out at 10 W.smo 5\u2019 UTR F and smo ATG R were used to detect truncations in smo 5\u2019 UTR.PCR for genotyping was performed using GoTaq (Promega). Quantification of mutagenesis efficiency of various gRNA was performed as described . BrieflyFor quantification of mutagenesis efficiency by cloning and sequencing, genomic DNA was extracted from twenty injected embryos and the gRNA target site was amplified by PCR (Smo 5\u2019 UTR F and Smo 5\u2019 UTR R) and cloned into pGEM-Teasy (Promega). Approximately twenty colonies were picked for colony PCR and amplified DNA was used for subsequent sequencing using Smo 5\u2019 UTR R.After injection of 250 fg Cas9 mRNA, embryos were shifted to 34.5\u00b0C when they reached 64 cell stage. Embryos were heat-shocked twice beginning at High stage for 2 h at 37\u00b0C with a 2 h interval at 34.5\u00b0C. After heat shock embryos were incubated overnight at 34.5\u00b0C.Hammerhead (HH) ribozymes are comprised of a RNA sequence motif that catalyzes self-cleavage from the 3\u2019 end of the RNA molecule and they have been used to remove 5\u2019 end heterogeneity of RNA transcripts. Cleavage of the HH ribozyme occurs at stem loop one . To makein-vitro transcribed (IVT) gRNA with ribozymes was able to guide Cas9 cleavage of its target in an in-vitro cleavage assay [in-vivo mutagenesis efficiency, a construct encoding HH-gRNA targeting the 5\u2019 UTR of mRNA encoded by the zebrafish smoothened (smo) gene was generated. IVT of smo HH-gRNA showed the presence of a band corresponding to gRNA after electrophoresis showed a dramatic loss of GFP expression in their slow-twitch muscle fibres, reflecting efficient mutagenesis of the GFP target site of each gRNA together with 200 fg of Cas9 mRNA. Previously it was shown that polyacrylamide gel electrophoresis (PAGE) was a reliable method for the detection as well as quantification of subtle allelic alterations [smo 5\u2019UTR. To determine the efficiency of mutagenesis, genomic DNA was extracted from single embryos and smo 5\u2019UTR was amplified by PCR. Efficiency of mutagenesis was determined by comparing the ratio of heteroduplex and homoduplex. We found that gRNA generated using ribozymes is as efficient as gRNA generated using conventional methods, both show ~60% efficiency that drives ubiquitous expression of Cas9 and mCherryHH-gRNA-HDV targeting GFP (Tg(smyhc1:GFP)i104 embryos and expression of H2a mCherry was monitored to confirm expression of gRNA. We found a number of embryos (n = 24) with mosaic expression of H2a mCherry showing a restricted loss of GFP expression in slow-twitch muscle fibres, suggesting that expression of the construct could induce mutation of its target site was generated expressing two gRNAs targeting Smo ATG and Smo 5\u2019 UTR driven by the heat shock promoter Sequencing results showing mutant alleles recovered from a pool of twenty injected embryos with the various gRNA constructs and Cas9 mRNA. gRNA target site is shown above with the PAM nucleotides shown in green. Red dashes and nucleotides in blue represent deletions and insertions respectively. Red line shows mutant clones after CRISPR/Cas9 mediated mutagenesis. (B) Table showing number of GFP+ slow twitch fibres remaining in WT, RNA of Cas9/GFP gRNA and Ubi: Cas9 Ubi: GFPgRNA injected embryos.(TIF)Click here for additional data file.S2 Fig\u03bcm.Embryos were heat shocked according to materials and methods section to test for expression of mCherry. Left panel show wild type embryos and right panel show HSP: Smo gRNA. Green asterisk show the expression of GFP in the heart from the cmlc: GFP used as the transgenesis marker. Embryos shown are 30 hpf. Scale bars: 500 (TIF)Click here for additional data file.S3 Fig\u03bcm. (B) Embryos were stained with Engrailed (Eng) to show defects in muscle specification when smo is mutagenesized. In the middle and right most panel, curved embryos were imaged and these embryos have defective formation of muscle pioneers and media fast fibers which are dependent on smo for its formation (Embryos shown are representative of the variation in observed phenotypes). Furthermore, these embryos have U-shaped somites (dotted line showing outline of somite in DAPI images) typical of lost of hedgehog signalling. Scale bars: 40 \u03bcm.(A) Overview of control embryos (uninjected HSP: Smo gRNA), embryos injected with Cas9 mRNA and Smo gRNA, and heat shocked HSP: Smo gRNA embryos injected with Cas9 mRNA. Scale bars: 500 (TIF)Click here for additional data file.S4 Fig(A) Workflow for the mutagenesis of zebrafish. If conditional mutagenesis is not the aim of the experiment, last two steps (marked with *) can be omitted. (B-C) Plasmid map and sequence of p3E U6 HH gRNA plasmid. This vector can be used for gateway cloning. Primers used for designing gRNA and template for IVT are shown (C). As mentioned before HH ribozyme requires complementary sequence to the gRNA target sequence (shown by yellow box).(TIF)Click here for additional data file.S1 File(DOCX)Click here for additional data file.S1 Table(DOCX)Click here for additional data file."} +{"text": "Does tranexamic acid lead to changes in MRI measures of brain tissue health in patients with spontaneous intracerebral haemorrhage? Protocol for a MRI substudy nested within the double-blind randomised controlled TICH-2 trial. BMJ Open 2018;8:e019930. doi: 10.1136/bmjopen-2017-019930Dineen RA, Pszczolkowski S, Flaherty K, On the secondary outcomes subsection (page 3) it reads:\u2018Only DWIHL that are confirmed of low diffusion on the derived apparent diffusion coefficient maps and spatially remote from the index ICH (<20 mm) will be included as previously.\u2019It should read:\u2018Only DWIHL that are confirmed of low diffusion on the derived apparent diffusion coefficient maps and spatially remote from the index ICH (>20 mm) will be included as previously.\u2019"} +{"text": "We thank Boucher et al. for theiTo directly answer their query regardinRegarding circulating parathyroid hormone (PTH) levels, these were not detectable in the serum of male C57BL/6J mice from the experiments described in , as meas"} +{"text": "Modulation of APE1 acetylation level in cells alters AP site repair capacity of the cell extracts in vitro. Primary tumor tissues of diverse cancer types have higher level of acetylated APE1 (AcAPE1) compared to adjacent non-tumor tissue and exhibit enhanced AP site repair capacity. Importantly, in the absence of APE1 acetylation, cells accumulate AP sites in the genome and show increased sensitivity to DNA damaging agents. Together, our study demonstrates that elevation of acetylation level of APE1 in tumor could be a novel mechanism by which cells handle the elevated levels of DNA damages in response to genotoxic stress and maintain sustained proliferation.Apurinic/apyrimidinic (AP) sites are frequently generated in the genome by spontaneous depurination/depyrimidination or after removal of oxidized/modified bases by DNA glycosylases during the base excision repair (BER) pathway. Unrepaired AP sites are mutagenic and block DNA replication and transcription. The primary enzyme to repair AP sites in mammalian cells is AP endonuclease (APE1), which plays a key role in this repair pathway. Although overexpression of APE1 in diverse cancer types and its association with chemotherapeutic resistance are well documented, alteration of posttranslational modification of APE1 and modulation of its functions during tumorigenesis are largely unknown. Here, we show that both classical histone deacetylase HDAC1 and NAD Mammalian apurinic/apyrimidinic (AP) endo nuclease 1 (APE1) is a ubiquitous and multifunctional protein. It plays a central role in the repair of AP sites generated either spontaneously or as an intermediate during the repair process of oxidative and drug-induced alkylation damage in the genome via the base excision repair (BER) pathway . APE1 clin vitro and in cells ATP using T4 polynucleotide kinase [2. The reaction was stopped with 10 \u03bcl 80% formamide/40 mM NaOH containing 0.05% xylene cyanol, followed by heating at 95\u00b0C for 5 min; the samples were kept on ice until ran in denaturing gel electrophoresis in 20% polyacrylamide containing 8 M urea to separate the substrate oligomer from the cleaved product. The gels were dried and the radioactivity was quantitated by phosphoimager analysis in a Storm system (Molecular Dynamics).A 43-mer oligonucleotide containing AP site analog tetrahydrofuran (THF) at nucleotide 31 (Midland Corp) prepared as described previously was 5\u2032-end-labeled with [\u03b3-e kinase , 76. FolAPE1siRNAHEK293T cells with Dox treatment [Endogenous APE1 was downregulated in Trevigen's FPG FLARE Comet assay kit was used following manufacturer's protocol. Briefly, control and GOx treated cells were lysed in Comet slides in low melting agarose followed by incubation with FPG before alkaline electrophoresis. DNA in the nucleoid was visualized by SYBR Gold staining in a fluorescence microscope . Data analysis was performed using Open Comet of Image J program (NIH) in 50-100 randomly selected cells and plotted as mean Tail moment.APE1siRNAHEK293T cells were treated with Dox for 5-6 days to knockdown endogenous APE1. Cells were transfected with WT or mutant APE1 expression plasmid. Colony formation assay was performed after treatment GOx as described above.HCT116 cells stably expressing APE1 shRNA were transfected with WT or mutant APE1 expression plasmid. After forty 48 hours of transfection, approximately 500 cells plated on 60-mm dishes were treated with various doses of methylmethane sulphonate for 1 hour, washed in PBS and allowed to grow in fresh medium for two weeks until visible colonies appear. HCT116 cells expressing control shRNA used as a control. The colonies were fixed with 100% methanol, stained with Giemsa staining solution (1:50) and counted. SimilarlyStatistical analysis was done using Student T Test and p value less than 0.05 was considered significant."} +{"text": "Enhanced Disease Susceptibility1 (EDS1) is a nucleo-cytoplasmic protein, known to be a key regulator of plant basal defense andeffector-triggered immunity. Sequence of a single copy brinjal EDS1 gene (SmEDS1) was mined from draft brinjal genome assembly.The extracted sequence was found to be incomplete and polished with the help of transcriptome sequence data. Full-length SmEDS1gene is 4.5kb long having three exons that coded for 1.8kb mRNA. SmEDS1 protein is a 602 amino acid long protein consisting ofLipase3 and EP domain regions. Predicted tertiary structure of SmEDS1 using homology modelling had a mass of 68.8kD and wasmade of 10 strands, 26 alpha helices, five 310 helices and 43 beta turns. Phylogenetic analysis based on protein sequence grouped thespecies in clades defined by botanical family suggesting that EDS1 protein has evolved through the speciation process. Phylogenetictree based on EDS1 structures grouped Solanum species of American origin together but brinjalEDS1 (Asiatic origin) occupied a unique position. In silico information generated in this study is expected to be the first step towardcloning and expression analysis of SmEDS1 gene. Effector triggered immunity (ETI), responsible for diseaseresistance in plants , is actiBrinjal (Solanum melongena L.) is an important solanaceousvegetable crop of Asiatic origin. Production of brinjal is affectedby diseases , nematodes and insects. As aresult, identification genes imparting biotic stress resistance inbrinjal is the main focus for varietal development and thedevelopment of genetically modified organisms. EDS1 could bean ideal target as it regulates both basal as well as effectortriggered defense pathways. As EDS1 is the key node of defenseregulatory system that functions at a very primary level, itsmodulation using CRISPR/Cas9 offers eIn the present study we report identification and characterizationof Solanum melongena EDS1 gene (SmEDS1) and propose itsprotein structure. We also present molecular phylogeneticanalysis based on EDS1 protein sequences and structures amongplant species. In silico information generated in this study is expected to be the first step toward cloning and expressionanalysis of SmEDS1.Brinjal genome was searSmEDS1 (602 amino acid) protein sequence was subjected toprotein structure modelling using Swiss-model, a homologymodelling web server . StructuAll available plant EDS1 sequences were collated by queryingNCBI protein database, EggNOG database and EnseOne representative from each clade of sequence-based NeighborJoining tree was taken for 3D structure-based phylogeneticanalysis based on structural comparison. Structure of each of theselected protein sequence was predicted by homology modelling. Structuralalignment of the modelled structures and reconstruction ofphylogeny was carried out using Multiseq .Mining the brinjal genome database using tblastn algorithm withAtEDS1 as the query sequence showed a homologue on thecontig Sme2.5_09498.1 . Ab initio gene predictionby FGENESH resulted in the gene coding for 486 amino acidswhereas homology based gene prediction by FGENESH+ [An N-terminal lipase_3 (PF01764.22) was predicted in Pfamresult. CDD results predicted Esterase-lipase superfamily domainin range 44-217. Search by protein sequence using blast algorithmin ECOD database displayed hit in N-terminal Lipase_3 domainand C-terminal EP domain. All these features were typical toEDS1 proteins. The promoter sequence analysis using the PlantCARE database indicated toward the presence of ABREelement (ABscisic acid Response Element), Box-4, G-box CATTmotifand sp1 (light-responsiveness), TC-rich repeats (defenseandstress-response) and HSE element (heat stress response).Tertiary structure of SmEDS1 was modelled using AtEDS1 crystalstructure (4NFU). The predicted structure was made of 10strands, 26 alpha helices, five 310 helices and 43 beta turns . The briPhylogenetic analysis of EDS1 protein sequences representing 49plant species using neighbor-joining tree grouped the speciesbelonging to the same family together in a single clade .Such a A common measure of structural similarity between twohomologous protein structures is root mean square deviation(RMSD) of topologically equivalent C-alpha atoms and used tofor phylogenetic analysis . The topIdentifying, annotating and characterizing EDS1 protein in cropslike brinjal, which is seriously affected by biotic stress, canprovide a novel way in resistance breeding. Availability ofscaffold-level genome sequence and new possibilities via geneeditingtechnologies motivated the identification andcharacterization of EDS1 in brinjal. Amino acid sequence basedphylogenetic grouping of plant EDS1 proteins exhibitedcongruence with taxonomy. This observation indicates theevolutionary and hence functional significance of EDS1 in plants.3D structure-based phylogenetic relationships on the other handrevealed no such analogy with genera or species. In silico analysishas revealed conservation of sequence motifs and structures to alarge extent among plant EDS1 proteins. SmEDS1 is surmised asan ideal candidate for genetic engineering experiments."} +{"text": "Pathological scores in the DEX were also similar in both groups. Correlations between PBCs and DEX scores were different in MS and control groups. MS patients with cognitive impairment had a marginally higher number of PBCs than control patients (p = 0.056) and a significantly higher DEXp score (p = 0.04). These results suggest that (1) PBCs occurring in early MS patients were not different from those induced by comparable chronic non-CNS disorders, (2) qualitative differences in the relationship between behavioral symptoms and executive-behavioral changes may exist between MS and control groups, and (3) behavioral symptoms seem associated with cognitive deficits in MS. We further plan to assess these observations longitudinally.Acquired behavioral changes have essentially been described in advanced multiple sclerosis (MS). The present study was designed to determine whether behavioral modifications specifically related to the MS pathological process could be identified in the initial phase of the disease, as compared to control patients with chronic, relapsing and progressive inflammatory disorders not involving the central nervous system (CNS). Eighty-eight early MS patients and 48 controls were tested. Perceived changes by informants in behavioral control, goal-directed behavior, decision making, emotional expression, insight and interpersonal relationships were assessed using the Iowa Scale of Personality Change (ISPC). Executive behavioral disturbances were screened using the Dysexecutive Questionnaire (DEX). The mean change between the premorbid and postmorbid ISPC ratings was similar in the MS [12.2 (SD 15.6)] and in the control [11.5 (SD 15.1)] group. The perceived behavioral changes (PBCs) most frequently reported in both groups were"} +{"text": "Scientific Reports 10.1038/s41598-017-11441-4, published online 08 September 2017Correction to: The original version of this Article contained an error in the bioinformatics pathway that resulted in the misalignment of OTU abundances with the incorrect corresponding OTU identities. In addition, the alignment database utilized was misreported as Greengenes instead of SILVA.As a result, in the Abstract,\u201cAscidians hosted diverse symbiont communities, consisting of 5,696 unique microbial OTUs (at 97% sequenced identity) from 47 bacterial and three archaeal phyla.\u201dnow reads:\u201cAscidians hosted diverse symbiont communities, consisting of 5,696 unique microbial OTUs (at 97% sequenced identity) from 44 bacterial and three archaeal phyla.\u201dIn the results section under subheading \u2018Symbiotic microbial community composition and diversity\u2019,\u201cAscidian-sourced OTUs spanned 42 bacterial and three archaeal phyla , with all archaeal phyla represented in all three ascidian species, though relative abundances varied among the different sources . Bacterial phyla likewise differed in relative abundance among the three species investigated . Microbial communities in D. bermudensis included 36 bacterial phyla and were dominated by Alphaproteobacteria (68%) and Gammaproteobacteria (7%), as well as Crenarchaeota . Symbiont communities in P. anguinea included 37 bacterial phyla and were dominated by unclassified Bacteria (34%), Alphaproteobacteria (27%), Gammaproteobacteria (18%) and Bacteroidetes . Microbial communities in P. zorritensis included 40 bacterial phyla and were dominated by Alphaproteobacteria (42%), Gammaproteobacteria (24%), Bacteroidetes (10%), and Planctomycetes .\u201dnow reads:\u201cAscidian-sourced OTUs spanned 44 bacterial and three archaeal phyla , with Euryarchaeota and Crenarchaeota represented in all three ascidian species, though relative abundances varied among the different sources . Parvarchaeota was detected only in the two Polyandrocarpa species, and only in low concentrations. Bacterial phyla likewise differed in relative abundance among the three species investigated . Microbial communities in D. bermudensis included 32 bacterial phyla and were dominated by Alphaproteobacteria (38%), Gammaproteobacteria (22%), unclassified bacteria (19%), and Bacteroidetes . Symbiont communities in P. anguinea included 39 bacterial phyla and were dominated by Alphaproteobacteria (66%) and Gammaproteobacteria (7%), as well as Crenarchaeota . Microbial communities in P. zorritensis included 42 bacterial phyla and were dominated by Alphaproteobacteria (47%), Gammaproteobacteria (17%), Bacteroidetes (9%), and Planctomycetes (5%), as well as Crenarchaeota .\u201dIn the Results section under subheading \u2018Core symbiont community composition and diversity\u2019,P. anguinea and 95.8% of unclassified sequences for all three species together.\u201d\u201cThis one unclassified OTU represented 98.6% of all unclassified bacterial sequences in now reads:D. bermudensis and 94.0% of unclassified sequences for all three species together.\u201d\u201cThis one unclassified OTU represented 98.3% of all unclassified bacterial sequences in In the Discussion section,39.\u201d\u201cIndeed, in the current study, most universal ascidian symbiont OTUs (80.3%) were also detected in the seawater, with three of these OTUs matching identically (100% pairwise identity) to sequences previously reported from seawater samples or sponge tissue within the same regionnow reads:39.\u201d\u201cIndeed, in the current study, most universal ascidian symbiont OTUs (80.3%) were also detected in the seawater, with some of these OTUs matching identically (100% pairwise identity) to sequences previously reported from seawater samples or sponge tissue within the same regionIn the Methods section under subheading \u2018Next-generation sequence data processing\u2019,89, sequences were classified using a naive Bayesian classifier and bootstrap algorithm for confidence scoring90 based on the improved Greengenes taxonomy91, and nontarget sequences and singletons were removed from the data set.\u201d\u201cBriefly, raw sequences were quality-filtered and aligned to the Greengenes reference database (gg_13_5_99), putative chimeric sequences were removed via self-reference searching with UChimenow reads:89, sequences were classified using a naive Bayesian classifier and bootstrap algorithm for confidence scoring90 based on the improved Greengenes taxonomy91, and nontarget sequences and singletons were removed from the data set.\u201d\u201cBriefly, raw sequences were quality-filtered and aligned to the SILVA reference database (v119), putative chimeric sequences were removed via self-reference searching with UChimeThese errors also affected Tables 2 and 3 and Figure 2 which have all been replaced.In the Supplementary Information file originally published with this Article, (Table S2) was inadvertently incorrectly included and has now been removed.These errors have now been corrected in the HTML and PDF versions of this Article, and in the accompanying Supplementary Material."} +{"text": "The telomere length distributions of MPNSTs were within the length-ranges in which telomere fusion is detected and that confer a poor prognosis in other tumor types. These data indicate that telomere length may play a role in driving genomic instability and clonal progression in NF1-associated MPNSTs.Neurofibromatosis type 1 is a familial cancer syndrome that affects 1 in 3,500 individuals worldwide and is inherited in an autosomal dominant fashion. Malignant Peripheral Nerve Sheath Tumors (MPNSTs) represent a significant cause of morbidity and mortality in NF1 and currently there is no treatment or definite prognostic biomarkers for these tumors. Telomere shortening has been documented in numerous tumor types. Short dysfunctional telomeres are capable of fusion and it is considered that the ensuing genomic instability may facilitate clonal evolution and the progression to malignancy. To evaluate the potential role of telomere dysfunction in NF1-associated tumors, we undertook a comparative analysis of telomere length in samples derived from 10 cutaneous and 10 diffused plexiform neurofibromas, and 19 MPNSTs. Telomere length was determined using high-resolution Single Telomere Length Analysis (STELA). The mean Xp/Yp telomere length detected in MPNSTs, at 3.282 kb, was significantly shorter than that observed in both plexiform neurofibromas (5.793 kb; [ NF1 tumor suppressor gene, located at 17q11.2. NF1 patients develop a variety of tumor types, including cutaneous neurofibromas, plexiform neurofibromas, malignant peripheral nerve sheath tumors, (MPNSTs), optic gliomas, phaechromocytomas, glomus tumors, gastrointestinal tumors and leukaemia labelled telomere repeat containing probe together with a probe to detect the 1 kb and 2.5 kb molecular weight markers. The hybridised fragments were detected by phosphorimaging with a Typhoon FLA 9500 phosphorimager . The molecular weights of the DNA fragments were calculated using the Phoretix 1D quantifier .19 macrodissected MPNSTs were obtained from 19 unrelated patients, in addition 10 diffused plexiform and 10 cutaneous neurofibromas were also taken from 20 unrelated NF1 patients. The samples used in this study were provided by genetic centers in Cardiff (UK), London (UK), Toronto (Canada) and Hamburg (Germany). The MPNSTs were classified in accordance with the WHO classification scheme and FNCLeviously . The DNAStatistical comparisons were made using the nonparametric Mann Whitney tests (Prism 6)."} +{"text": "The most common adverse events associated with Thymoglobulin are cytokine release syndrome, thrombocytopenia, and lymphopenia. Results of early studies showed an increased rate of cytomegalovirus disease associated with Thymoglobulin treatment, but recent studies indicate that routine administration of modern antiviral prophylaxis can reduce this risk. More research comparing Thymoglobulin with basiliximab will help individualize regimens by matching the choice of induction agent with the risk profile of each transplant recipient. The proven efficacy and safety profile of Thymoglobulin provides an excellent starting point for future investigations.[38]Horse ATG (hATG) or Thymoglobulin + Cyclosporine are an efficacious treatment for aplastic anemia. Due to its higher potency Thymoglobulin may be superior to hATG, but further studies are required for confirmation. and for treatment of aplastic anemia.Attributable to its polyclonal nature, Thymoglobulin provides multifaceted immunomodulation suggesting that its use should be included in the immunosuppressant therapeutic armamentarium to help reduce the incidence of organ rejection and GvHD,[ Immunosuppressive properties of polyclonal antithymocyte globulins (ATG) were first described in the 1950s,[4] Currently there are three different ATGs commercially available: Human thymocytes are used as the immunogenic to produce Atgam\u00ae in horses and Thymoglobulin\u00ae in rabbits, respectively; a Jurkat cell line is used to produce ATG-Fresenius\u00ae in rabbits.[4] Despite sharing some common properties, the commercially available ATG products are strictly different drugs.[5] Immunosuppressive activity varies significantly from one preparation to the other, resulting in quite different dosages. Among these products, Thymoglobulin is probably the most potent, and the most extensively studied ATG. This review describes the clinical use of Thymoglobulin in organ transplantation and hematology/oncology.ATG is a mixture of different antibody specificities, which induces an extremely effective dose-dependent T-cell depletion in blood and lymphoid tissues via complement-dependent cytotoxicity, antibody dependent cellular cytotoxicity, and apoptosis.[7]The role of Thymoglobulin in the prevention and treatment of allograft rejection, graft versus- host disease (GVHD), and treatment of aplastic anemia (AA) is well established. Recent investigations have shown that Thymoglobulin does not only deplete T-cells, but modulates various lymphocyte surface antigens and interferes with the function of a number of different immune effector cells, including B cells, dendritic cells, natural killer (NK) T cells, and regulatory T cells (Tregs).[8] Thymoglobulin is indicated for prevention of graft rejection in organ transplantation (induction); dosage 1 to 1,5 mg/kg/day for 2 to 9 days (2 to 5 days in heart transplantation).[9] In the US, antibody induction is used in the majority (>70%) of kidney and almost 50% of thoracic organ transplantations, and Thymoglobulin is the most frequently used induction agent.[10] It has the following roles in organ transplant recipients: reduction of the incidence of acute rejection, prevention of ischemia reperfusion injury and delayed graft function, and minimization of calcineurin inhibitors (CNIs) and/or corticosteroids. The risk of organ rejection is bigger immediately (weeks to months) after transplantation. It declines during the first year and further on, but it is present through the whole life of the graft. A retrospective analysis in living donor kidney transplantation (n=214) in a single center versus a national cohort showed a significant benefit of Thymoglobulin induction vs. no antibody induction in a low risk patient population. Five years patient survival was 96% vs. 90% , and acute rejection at one year was 2% vs. 21% . Thymoglobulin was well tolerated with very few infections, and a low incidence of malignancy.[13]In two randomized, prospective trials Thymoglobulin was shown to decrease the rate of acute rejection in kidney transplant patients compared to no induction . In these early studies, the incidences of leucopenia, thrombocytopenia, fever, and cytomegalovirus infection were significantly higher in the Thymoglobulin groups.[14] In a prospective, double blind trial event free survival after one , five , and ten years was significantly higher in Thymoglobulin treated patients (n=48) receiving a kidney transplant compared to Atgam (n=24). There were no post transplant lymphoproliferative disorder (PTLD) in the Thymoglobulin group and two cases in the Atgam group.[15] In a single-center, retrospective study Thymoglobulin induction (n=65) seemed to be connected with higher rates of CMV disease, malignancy, and death than ATG Fresenius (n=129).[16] However limitations of this study are the variable doses of Thymoglobulin ,[4] and lack of antiviral prophylaxis for the longest period of time.One prospective, randomized trial compared induction with Thymoglobulin (n=28) and ATG Fresenius (n=30) in kidney transplant recipients. Acute rejection after one year was numerically lower in the Thymoglobulin group . Thymoglobulin patients experienced a lower incidence of infections, lower white blood cell (WBC) counts while maintaining hemoglobin levels better.[17]A retrospective analysis compared the long-term benefits of induction with Thymoglobulin (n=342) and ATG Fresenius (n=142) in heart transplantation from 1984 to 1996. Five year patient survival rate was significantly higher in the Thymoglobulin group ; 72% versus 42% of patients were free from acute rejection, less and less severe recurrent rejections were observed. Viral infections , but not cytomegalovirus (CMV) infections (17% vs. 13%), were more frequently observed in the Thymoglobulin group. Post transplant lymphoproliferative disorders (PTLD) were comparable. The authors concluded, that the two rabbit ATGs have different immunosuppressive potency, and that Thymoglobulin is currently the most powerful induction agent in heart transplantation.[18] and five years in the US cohort (15% vs. 25%).[19] Moreover acute rejection following Thymoglobulin induction was less severe: Rejection rates requiring antibody treatment were 1,4% vs. 8% at one year[18] and 3% vs. 12% in the US cohort after five years .[19] The authors concluded, that Thymoglobulin and basiliximab have equivalent but different safety profiles, and require appropriate antibacterial and antiviral prophylaxis strategies.[18] CMV disease occurred more frequently in patients treated with basiliximab , whereas the rate of infections was higher in the Thymoglobulin group .[18] However an economic analysis showed, that in Thymoglobulin patients 12 months post-transplant, treatment costs were continuously lower. The combination of lower costs and improved outcomes make Thymoglobulin both clinically and economically preferred over basiliximab in patients receiving a kidney from a marginal donor.[20]In a prospective, randomized trial comparing induction with Thymoglobulin (n=141) and basiliximab in patients receiving a kidney transplant from a marginal donor, BPAR was lower in the Thymoglobulin group after one [21] Both studies showed no statistical difference between Thymoglobulin and anti-IL-2R antibodies in terms of patient- and graft survival.In another prospective randomized trial comparing Thymoglobulin (n=113) and daclizumab in high risk renal transplant patients with triple maintenance therapy, BPAR after one year was significantly lower in the Thymoglobulin group .[22] Steroids and CNI have been cornerstones for the maintenance of immunosuppression, but are associated with side effects affecting graft and patient survival and the quality of life.[22]Thymoglobulin induction has proven success in CNI and steroid minimization strategies. Today, death with a functioning graft and chronic allograft nephropathy (CAN) are major causes for late graft loss.[23]A prospective study (n=150) showed excellent results of Thymoglobulin induction in a CNI\u2013free maintenance and steroid-tapering protocol. Patients were randomized to either a sirolimus- based or a cyclosporine A (CsA) -based regimen. All patients received mycophenolate mofetil (MMF) and a 6-month course of corticosteroids. At the 12-months follow-up, 88% of patients were steroid free. No significant differences were observed in patient survival (97% in each treatment group), graft survival (90% vs. 93%) or acute rejection (14.3% vs. 8.6%).[24]In a retrospective study of cadaveric renal transplant recipients treated with Thymoglobulin or basiliximab (high and low risk n=115), maintenance with sirolimus and prednisone, and delayed introduction of reduced-dose CsA, BPAR at 3 months in high-immune responders was lower in patients receiving Thymoglobulin . Serum creatinine was higher with basiliximab at 3, 6, and 12 months (p<0.02). Only Thymoglobulin showed an excellent result when CsA initiation was delayed for more than two weeks (0% vs. 24%).[25]A single center trial of prednisone-free maintenance immunosuppression using Thymoglobulin induction and CsA/MMF or tacrolimus/sirolimus (TAC/SRL) in 589 patients showed an excellent five year patient and graft survival (91% and 84%), low incidence of acute rejection, and stable kidney function . Steroid-related side effects like CMV infection and post transplant diabetes were minimized compared to the historic controls . Thymoglobulin induction plus elimination of prednisone should be considered at least in low risk recipients.[26]Treatment of acute rejection requires a short course of more intensive immunosuppressive therapy. First-line therapy for acute rejection is usually high dose intravenous corticosteroids. In case of steroid resistant acute rejection alternative treatments are necessary, which can be either ATG or the monoclonal antibody muromonab-CD3 .[9]Thymoglobulin is indicated for the treatment of graft rejection in organ transplantation with a recommended dosage of 1,5 mg/kg/day for 3 to 14 days.[27] Due to less frequent treatments of recurrent rejection and less frequent return to dialysis, Thymoglobulin provided significant cost savings.[28]A double-blind randomized trial showed superiority of over Atgam in reversal of acute rejection and prevention of recurrent rejection in patients who received a renal transplant. Both drugs had a similar side effect profile. The enhanced clinical efficacy of Thymoglobulin was explained by higher affinity of rabbit IgG subtype to human lymphocytes, less batch-to-batch variability, longer half-life, and more profound and longer lasting lymphocyte depletion compared to horse ATG.[29] Similar efficacy of ATG or OKT 3 treatment and a fewer side effects related to first dose syndrome were reported in a meta-analysis of 21 clinical trials.[30]A randomized clinical trial comparing Thymoglobulin (n=31) and OKT 3 (n=29) in treatment of steroid resistant acute rejection in kidney transplant patients showed a trend in favor of Thymoglobulin . Fever, mainly due to first dose syndrome, occurred more frequently in the OKT 3 group .[31] is a rare, potentially life-threatening failure of haemopoiesis characterized by pancytopenia and bone marrow aplasia.[32] Most cases of AA are acquired, although very rare inherited forms exist.[32] Acquired AA can occur in any age group, and in most cases results from an autoimmune attack against hematopoietic stem cells.[31] Immunosuppressive therapy with ATG + Cyclosporine is treatment of choice in patients above the age of 50 years, for patients who lack an HLA identical donor, and for patients with non-severe aplastic anemia.[33] Thymoglobulin is indicated for the treatment of aplastic anemia; recommended dosage between 2,5 and 3,5 mg/kg/day for 5 consecutive days.[9] For historical reasons most European studies in AA have been carried out using Lymphoglobuline\u00ae , a horse ATG from the same manufacturer, which was available on the market 16 years earlier than Thymoglobulin.Aplastic anemia (AA), the paradigm of human bone marrow failures,[34] First line use of Thymoglobulin is recommended in the British Committee for Standards in Hematology (BCSH) guidelines for AA, because Lymphoglobuline is no longer available.[35] Lymphoglobuline (horse ATG) and Thymoglobulin source of antigen has been identical, both derived from human thymocytes and having comparable biological effects. Two clinical trials have shown similar response rates for both ATG in bone marrow failure. Thymoglobulin shows excellent response and good safety profile in first line treatment of SAA[38] and has significant activity (33% and 42%) in low-risk MDS. Retreatment with Thymoglobulin in patients not responding to a first course with Lymphoglobuline was associated with excellent response (77%) and survival rates (93% at 912 days follow up), and without relapse.[40]The current EBMT guidelines recommend either horse or rabbit ATG in first line treatment of AA. may be due to its different antigen source (Jurkat cells).[42]The unsatisfactory result of ATG-F in treatment of aplastic anemia (response rates between 47% and 53%) Thymoglobulin is indicated for prophylaxis of acute and chronic graft versus host disease (GvHD), after hematopoietic stem cell transplantation; dosage is 2,5 mg/kg/day from day -4 to day -2 or -1.[9]Both acute (a) and chronic (c) graft versus host disease (GvHD) are a major cause of transplant related morbidity and mortality after allogenic hematopoietic stem cell transplantation (HSCT). ATG have been used to reduce the risks of graft failure and GvHD.[43] Thymoglobulin conditioning reduced cGvHD vs. no ATG and extensive cGvHD at 5 years follow up of a randomized trial, resulting in less chronic lung dysfunction and improved quality of life, measured as Karnofsky scores \u2265 90% at 4 years .[44] A retrospective analysis compared Thymoglobulin (n=49), ATG Fresenius (n=38) and no ATG (n=68). ATG had a positive effect on cGvHD vs. no ATG. The better leukemia-free survival and low rates of relapse with Thymoglobulin outweighs the higher incidence of cGvHD in the Thymoglobulin group compared with ATG Fresenius.[45]Several trials were performed in myeloablative conditioning regimens. A matched cohort study in patients receiving HSCT from matched unrelated donors (MUD) showed a benefit of Thymoglobulin conditioning (n=52) vs. no ATG (n=104) in transplant related mortality (TRM) =0,30; p=0,005) and overall survival . The risk of relapse was similar in both groups.[46] Low dose Thymoglobulin conditioning in matched related donors (MRD) vs. no ATG (n=54 matched pairs) resulted in lower non-relapse mortality and better overall survival despite the increased relapse rate after 4 years. PTLD and infections related to Thymoglobulin were not observed.[47]A single center comparison of patients undergoing allogenic HSCT from either HLA identical sibling (n=121) without or matched unrelated donor (MUD) (n=61) with Thymoglobulin conditioning showed similar survival (60% in both groups) and relapse at five years.[48] In a large retrospective analysis a regimen of fludarabine-busulfan (Flu Bu) Thymoglobulin showed the best long term outcome compared to Flu Bu regimens with different ATG doses or Flu-total body irradiation (TBI).[49]Reduced intensity conditioning (RIC) protocols aim to achieve both durable donor stem cell engraftment and reduced transplant related mortality (TRM) allowing allogenic HSCT in elderly and comorbid patients. A regimen of total lymphoid irradiation plus Thymoglobulin was shown to decrease acute GvHD and to allow graft anti-tumor activity in elderly patients (n=37).["} +{"text": "The present study evaluated the microbiota of saliva and tooth biofilm by sequencing segments of the 16S rRNA gene using the common Illumina MiSeq (v3\u2013v4) and the newer PacBio (v1\u2013v8) sequencing platforms and searched for associations with caries in a low-caries population with regular dental care since early childhood. Saliva and tooth biofilm from adolescents and mock bacteria communities were analysed, including validity and reliability estimates. Caries was scored at baseline and 2 years later. The two sequencing platforms revealed similar microbiota patterns for saliva and tooth biofilm, respectively. Saliva microbiota discriminated caries-affected adolescents from caries-free adolescents with enumeration of five species in caries-affected participants and one species thereof predicting 2-year caries increment. Reliability was high and validity was acceptable if appropriate inclusion criteria were determined."} +{"text": "Alveolar rhabdomyosarcoma (ARMS) represents a block in differentiation of malignant myoblasts. Genomic events implicated in the pathogenesis of ARMS involve PAX3-FKHR (FOXO1) or PAX7-FKHR (FOXO1) translocation with corresponding fusion transcripts and fusion proteins. Commonalities in ARMS include uncontrollable proliferation and failure to differentiate. The genomic-molecular correlates contributing to the etiopathogenesis of ARMS incorporate PAX3-FKHR (FOXO1) fusion protein stimulation of the IGF-1R, c-Met and GSK3-\u03b2 pathways. With sequential morphoproteomic profiling on such a case in conjunction with personalized tumor graft testing, we provide an expanded definition of the biology of PAX3-FKHR (FOXO1) ARMS that integrates genomics, proteomics and pharmacogenomics. Moreover, therapies that target the genomic and molecular biology and lead to tumoral regression and/or tumoral growth inhibition in a xenograft model of ARMS are identified.This case study could serve as a model for clinical trials using relatively low toxicity agents in both initial and maintenance therapies to induce remission and reduce the risk of recurrent disease in PAX3-FKHR (FOXO1) subtype of ARMS. Alveolar rhabdomyosarcoma (ARMS) is a highly aggressive soft tissue sarcoma affecting children and adolescent age groups. At the genomic level it can be associated with translocations involving t2;13) or t resulting in fusion transcripts of PAX3-FKHR (FOXO1) and PAX7-FKHR (FOXO1) genes and corresponding proteins, reportedly accounting for 55% and 22% of ARMS, respectively in one study ) expression in the cytoplasm of virtually all of the tumor cells at 1-3+ signal intensity x100. Individual mice reporting a tumor volume >120% of the Day 0 measurement are considered to have progressive disease (PD). Individual mice with neither sufficient shrinkage nor sufficient tumor volume increases are considered to have stable disease (SD). Individual mice reporting a tumor volume \u226470% of the Day 0 measurement for two consecutive measurements over a seven day period are considered partial responders (PR). If the PR persisted until study completion, percent tumor regression (%TR) is determined using the formula: %TR=(1-Tf/Ti)x100; a mean value is calculated for the entire treatment group. Individual mice lacking palpable tumors for two consecutive measurements over a seven day period are classified as complete responders (CR). All data collected in this study was managed electronically and stored on a redundant server system. The clinical specificity for this test is 60%; the clinical sensitivity of this test is 98.1%.At study completion, percent tumor growth inhibition (%TGI) values were calculated and reported for each treatment group (T) versus control (C) using initial (i) and final (f) tumor measurements by the formula:"} +{"text": "Fabry Disease (FD) is characterized by globotriaosylceramide-3 (Gb3) accumulation in several tissues and a small fibre neuropathy (SFN), however the underlying mechanisms are poorly known. This study aimed to: 1) ascertain the presence of Gb3 deposits in skin samples, by an immunofluorescence method collected from FD patients with classical GLA mutations or late-onset FD variants or GLA polymorphisms; 2) correlate skin GB3 deposits with skin innervation.we studied 52 genetically-defined FD patients , 15 patients with SFN associated with a specific cause and 22 healthy controls. Subjects underwent skin biopsy to evaluate Gb3 deposits and epi-dermal innervation.Skin Gb3 deposits were found in all FD patients with classical GLA mutations but never in FD patients with late-onset variants or GLA polymorphisms or in patients with SFN and healthy controls. Abnormal deposits were found inside different skin structures but never inside axons. FD patients with GB3 deposits showed lower skin innervation than FD patients with late-onset variants or polymorphisms.1) Skin Gb3 deposits are specific to FD patients with classical GLA mutations; 2) Gb3 deposits were associated with lower skin innervation but they were not found inside axons, suggesting an indirect damage on peripheral small fibre innervation. Fabry Disease (FD) is a rare x-linked disorder characteThe possibility to disclose skin Gb3 deposits is interesting because skin is an easily accessible tissue. Skin Gb3 deposits have been demonstrated by using complex techniques such as electron microscopy but theySpecific aims of this study were: 1) to ascertain the presence of Gb3 deposits in skin samples collected from FD patients with classical GLA mutations or late-onset FD variants or GLA polymorphisms by an immunofluorescence method; and 2) to ascertain the correlation of skin Gb3 deposits with the skin innervation.We studied 52 genetically-defined FD patients including 14 males and 38 females who belong to 26 different families Tables . To testThe procedures used were approved by the Human Ethics Committee of Policlinico Sant\u2019Orsola-Malpighi (Bologna) and followed the Helsinki Declaration regarding international clinical research involving human beings. All subjects gave their written informed consent to the study.Alpha-GAL activity was tested in 39 patients; the remaining 13 patients performed only the genetic analysis because alpha-GAL activity was thought to be a non-diagnostic priority since patients belong to a family with an FD mutation. In addition, this test was performed in several laboratories with different normative values. To obtain a uniform value among different patients, alpha-GAL activity was normalized to the lower cut-off value .2[All patients performed a neurological examination. Pain was evaluated by means of a visual analogue scale (VAS), scale marking the level of pain on a 100 mm, non-hatched scale which reported at one end \u201cno pain\u201d and at the other \u201cworst pain imaginable\u201d. A stand2. Cardiom2.Three mm punch biopsies were taken from proximal (thigh: 15 cm above the patella) and distal hairy skin sites. We were unable to take skin samples from the thigh in 6 patients because of patient\u2019s refusal. According to previously published procedures skin samFifty \u03bcm sections were obtained using a freezing sliding microtome to evaluate Gb3 deposits. Usually 3 free-floating sections were analysed for each skin sample. They were immunostained overnight with a panel of primary antibodies including mouse monoclonal anti-Gb3 , rabbit pan-neuronal marker protein gene product 9.5 and rabbit polyclonal anti-collagen IV . Sections were then washed and secondary antibodies were added for an overnight incubation. As secondary antibodies, an anti-rabbit Alexa Fluor(R) 488 or mouse cyanine dye fluorophores 3.18 were used. ULEX europaeus , a biotinylated endothelium binding lectin, labeled with cyanine dye fluorophore 5.18 coupled with streptavidin was used to show the endothelium. Sections were viewed and analysed under a Nikon confocal microscopy (Eclipse Ti A1). Each image was collected in successive frames of 1\u20132 \u03bcm increments on a Z-stack plan at the appropriate wavelengths for secondary antibodies with a x40 or x60 plan apochromat objective and subsequently projected to obtain a triple-stained 3D digital image by a computerized system. The microscope settings were kept the same for all cases analysed. The analysis was made in a blinded fashion by two authors with expertise in immunoflorescent analysis (DV and IA). Full agreement on the Gb3 staining was reached by the two analysers in all cases without any discordance or uncertain classification . Gb3 staining was rated in each skin site as the percentage of skin structures showing a positive staining at high magnification (x40).Additional 50 \u03bcm sections from the same skin sample were obtained during the freezing sliding microtome session. Three free-floating sections were incubated overnight with a panel of primary antibodies, including the rabbit pan-neuronal marker protein gene product 9.5 and mouse collagen IV to define the basal membrane dividing epidermis from dermis. Sections were then washed and secondary antibodies, labeled with mouse Alexa Fluor(R) 488 and rabbit cyanine dye fluorophores 3.18 were added for an overnight incubation. Epidermal nerve fibre density (ENFs: number of unmyelinated fibres per linear millimetre of epidermis) was calculated by considering a single epidermal nerve fibre marked by PGP 9.5 crossing the dermal\u2013epidermal junction.We performed statistical analyses using SPSS 15.0 for Windows. Parametric tests were used as Kolmogorov\u2013Smirnov test showed that the variables were normally distributed. Unpaired Student\u2019s t test was used to compare clinical and demographic data between: 1) FD patients vs controls or SFN patients; 2) FD subgroups . Linear correlation using Pearson test was used to correlate skin Gb3 deposits with clinical parameters or innervation scores. p<0.05 was considered significant.Neurological examination was essentially normal in FD patients except for 2 male patients showing distal tactile sensory loss with a sock distribution. Pain was reported by 21 FD patients, more by males than by females . Pain was more frequent in patients with classical mutations (66% vs 50%). Autonomic symptoms were reported by 22 patients and mainly included palm and sole sweating loss (19 patients) and gastrointestinal dysmotility with nausea, bloating, abdominal cramps or diarrhoea (6) primarily in the group of classical mutations (53% vs 25%). Fifteen patients, mainly those with non-classical mutations (35% vs 20%) had cardiomyopathy. Ten patients showed groin and umbilicus angiokeratomas. Three patients showed renal failure with decreased GFR and 19 pAbnormal deposits were found in all patients with classical mutations, whereas all patients with non-classical mutations were negative. FD patients positive for deposits showed a significant young age compared to negative patients . No cleaSkin Gb3 deposits were found in blood vessel walls and endothelial cells, sweat gland tubules, perineurial, arrector pilorum muscle and dermal cells but never inside the axons . AbnormaENFs was decreased in FD patients both in the leg and thigThe main conclusions of our study were: 1) skin Gb3 deposits are specific to FD patients with classical GLA mutations; and 2) Gb3 deposits were associated with lower skin innervation but they were not found inside the axons, suggesting an indirect damage on peripheral small fibre innervation.FD diagnosis is difficult due to non-specific onset of symptoms. Abnormal skin Gb3 deposits have been studied by complex and expensive techniques involving the use of electron microscopy althoughSFN is a typical finding in FD,20,22 anOur data are in line with these conclusions since we did not disclose Gb3 deposits in skin axons, which were found mainly in the skin vessel walls, sweat gland ducts or arrector pilorum muscle cells. These findings supported an indirect damage of Gb3 on skin\u2019s small nerve fibres. In addition, the ENFs lower value of the leg compared to the thigh regardless of a similar Gb3 deposition could suggest a length-dependent dysfunction as the underlying pathogenetic mechanism affecting small nerve fibres. However, the abnormal expression of ion channels in peripheral nociceptors could be involved in the neuropathic pain in the mouse model of FD also contributing to their structural damage, although the mechanism inducing these abnormalities is not completely understood,30.Finally, the wide range of intrafamilial phenotypic variability regarding the severity of the disease but also the affected target-organs in patients with the same GLA mutation should aOur study has the following limitations: 1) alpha-GAL activity was lacking in several patients preventing us from establishing a reliable correlation with ERT, skin innervation or Gb3 deposits. To this end, an additional study involving a large cohort of patients would be useful; 2) we analysed few male patients with non-classical mutations. An analysis of these patients will be helpful in the future to confirm our current data; 3) furthermore, the quantification of Gb3 deposits by an ultrastructural investigation may increase the accuracy of this analysis."} +{"text": "Arracacha virus A from a Peruvian arracacha sample collected in 1975 and compare it with the genomes of other nepoviruses. Its RNA1 and RNA2 both had greatest amino acid identities with those of the subgroup A nepovirus Melon mild mottle virus.We present here the first complete genomic sequence of Arracaccia xanthorhiza), family Apiaceae] showing pronounced leaf mosaic symptoms. The virus was mechanically transmissible to 38 species from 10 plant families. Following characterization using electron microscopy, density centrifugation, and serology, it was identified as a member of the Nepovirus genus and named Arracacha virus A (AVA) . In 1978In 2015, total RNA was extracted from the freeze-dried AVA-infected leaf material using an RNeasy kit , including the optional DNase treatment. An indexed plant ribosome-subtracted sequencing library was then produced from the total RNA using the ScriptSeq complete plant leaf kit , according to the manufacturer\u2019s instructions. The indexed library was then sequenced along with others on a MiSeq instrument (Illumina), using a 600-cycle V3 kit. The resulting 492,656 paired reads were trimmed on the 3\u2032 end to a quality score of 20 using Sickle (Melon mild mottle virus (39%) (Grapevine fanleaf virus (36%) (Melon mild mottle virus and Potato black ringspot virus (Nepovirus genus.The AVA RNA1 encoded a putative 266-kDa polyprotein with an amino acid identity resembling those of the subgroup A nepoviruses us 39%) and Grapus (36%) . The AVA% and GrKY569301 (RNA1) and KY569302 (RNA2).The sequences were deposited in GenBank under accession numbers"} +{"text": "Cell viability decreases induced by H2O2 were mitigated by different CP extracts using various solvents. From these active fractions, six active compounds were separated and identified. Among tested isolated compound, the cytoprotective activities of three caffeates, caffeic acid phenethyl ester (CAPE), benzyl caffeate (BZC), and cinnamyl caffeate (CNC), exerted stronger effects than chrysin, pinobanksin, and 3,4-dimethoxycinnamic acid (DMCA). These three caffeates also increased H9c2 cellular antioxidant potential, decreased intracellular calcium ion ([Ca2+]i) level, and prevented cell apoptosis. Overall, the cardiovascular protective effects of the CP might be attributed to its caffeates constituents and provide evidence for its usage in complementary and alternative medicine.Chinese propolis (CP) is known as a health food but its beneficial effects in protecting cardiomyocytes remain elusive. Here, we investigated the effects of CP and its active compounds on hydrogen peroxide (H Cardiovascular disease (CV) is responsible for 30% of deaths worldwide, surpassed other diseases, and is projected to account for 25 million deaths annually by 2030 . The cosApis mellifera) from buds of plants i were determined in H9c2 cells (\u03bcM) pretreatment significantly brought back myocardial ionizable calcium to near positive quercetin control. These data are consistent with a previous study in human endothelial cells (HUVEC) that cytosolic [Ca2+]i were also increased by CAPE i pool.Myocardial ischemic damages can be affected by apoptosis through at least three potential mechanisms: (1) reducing myocardial cell numbers, which will directly decrease the heart pumping function; (2) damaging the heart's conduction function; and (3) initiating myocardial remodeling and inducing other cardiac pathological changes , 35. ForIn summary, our study provides an important basis for the use of Chinese propolis for the prevention and treatment of cardiovascular diseases. The crude Chinese propolis extract as well as its isolated compounds caffeate derivatives could mostly possibly be useful for the development of new antimyocardial ischemia drugs, depending on their in vitro activity. However, further in vivo pharmacological and toxicity studies are necessary for its potential clinical usages.Thin Layer Chromatography (TLC) maps of different extracts (S1) and combined fraction extracts of Chinese propolis."} +{"text": "The novel SH3 domain protein Dlish/CG10933 mediates fat signaling in Drosophila by binding and regulating Dachs. Published 3, October 2016fat mutant wing disc.We discovered an error in The corrected The originally published The article has been corrected accordingly."} +{"text": "Rhegmatogenous retinal detachment associated with choroidal detachment (RRDCD) is a special type of complex retinal detachment, and usually has a poor prognosis. This study aimed to assess the anatomical outcomes of 23-gauge pars plana vitrectomy (23G PPV) combined with phacoemulsification (phaco) and capsulotomy without intraocular lens (IOL) implantation in patients with RRDCD.Seventy-six consecutive patients with RRDCD, who underwent retinal repair surgery from January 2010 to December 2014, were retrospectively analyzed. Forty patients underwent 23G PPV + phaco + IOL implantation, and 36 underwent 23G PPV + phaco + capsulotomy without IOL implantation . All cases were filled with silicone oil. The follow-up time was 6 months after silicone oil was removed. Multivariate logistic regression analysis was the statistical method used.P = .007).The overall retinal anatomical reattachment rate was 58% (44/76): 40% (16/40) of patients receiving 23G PPV + phaco + IOL implantation; and 78% (28/36) of patients receiving 23G PPV + phaco + capsulotomy + aphakia (Surgical repair using 23G PPV + phaco + capsulotomy without IOL implantation can improve anatomical reattachment rates in patients with RRDCD. Scleral buckling alone has been reported in 35% to 62% of cases.\u20133 Primary vitrectomy results in a higher rate of retinal reattachment, reported to be as high as 77%.\u20136 Studies have been performed to investigate the outcomes of vitrectomy combined with other surgical techniques. It is reported that pars plana vitrectomy (PPV) combined with scleral buckling improved the reattachment rate in RRDCD.,8 By contrast, however, a European multicenter retrospective study suggested that a supplemental buckle may not be helpful in RRDCD. Some surgeons suggested routine lensectomy, or intraocular lens (IOL) removal and capsulotomy in complex retinal detachment.,11 Nevertheless, few studies investigating capsulotomy in patients with RRDCD have been performed. Accordingly, we retrospectively observed 76 Chinese patients with RRDCD in our hospital from 2010 to 2014, and analyzed the efficacy of 23-gauge (23G) PPV (23G PPV) combined with phacoemulsification (phaco) and capsulotomy without IOL implantation on anatomical retinal reattachment rates.The incidence rate of rhegmatogenous retinal detachment (RRD) associated with choroidal detachment (RRDCD) is reported to be 2.0% to 4.5%.2A retrospective review including 76 patients who met the following inclusion criteria was performed: RRD with identification of breaks; presence of choroidal detachment (CD) diagnosed preoperatively using ocular ultrasound; proliferative vitreoretinopathy (PVR) with grades C or D ; operation(s) performed by 1 surgeon; and silicone tamponade. Patients who experienced trauma, tumor(s), or exudative disease, or who underwent previous vitrectomy or phaco were excluded. The study was approved by the hospital's ethics committee.All cases received the same pre- and postoperative treatments. A sclerotomy, through which the choroidal fluid had drained, was performed and subsequently re-entered using a 23G blade to penetrate the pars plana canals. 23G PPV + phaco + IOL implantation, or 23G PPV + phaco + capsulotomy without IOL implantation , was performed. Membrane peeling or retinectomy was performed if needed. Iridotomy was performed before silicone oil tamponade in cases with aphakia. All cases were treated with intraocular laser around retinal breaks and filled with silicone oil. All patients were followed-up for 6 months after silicone oil removal. Unsuccessful cases included: failure of retinal detachment repair; remaining silicone oil at study conclusion; or need for additional procedures to repair detachments.P\u200a<\u200a.05 was considered to be statistically significant.Multivariate logistic regression analysis Tables and 2, c33.1Seventy-six eyes of 76 patients met the inclusion criteria. The mean age of the patients was 56 years (range 17\u201378 years). Sixteen patients were high myopia, and 2 had a macular hole. Basic demographic information is summarized in Table 3.2P\u200a=\u200a.007) ; 36 received 23G PPV + phaco + capsulotomy + aphakia, with a reattachment rate of 78% (28/36) than 23G PPV + phaco + IOL implantation (40%). In contrast to patients with RRD, intravitreous inflammatory mediators are upregulated in patients with RRDCD, who usually experience more severe PVR. The overall retinal anatomical reattachment rate after the first primary vitrectomy in our study was only 58%, which was low. However, the 23G PPV + phaco + capsulotomy without IOL implantation had a higher reattachment rate, which was approaching to rates previously reported in the literature.\u20136 Vitrectomy and capsulotomy may improve anterior and base vitrectomy, which may decrease anterior PVR.,14 A study by Tseng et al reported that pseudophakic eyes exhibited a higher proportion of hypotony after retinal reattachment surgery than eyes without IOL implant. The lens capsule may fibrose and contract, causing secondary contraction of the ciliary body, which in turn leads to chronic hypotony.,18 Because RRDCD is usually accompanied by hypotony, possible advantages of capsulotomy include a lower hypotony rate after surgery, which may improve the reattachment rate in RRDCD. And the results of our study support the points.Retinal detachment recurrence is primarily due to PVR.Our study had several limitations. No further retinal reattachment rates were assessed after successive surgeries because some patients who experienced retinal detachment recurrence refused additional treatment. Visual outcomes\u2014which may be as important as anatomical outcomes\u2014were not assessed. Further prospective studies involving patients with RRDCD are required.5Surgical repair using 23G PPV + phaco + capsulotomy without IOL implantation can improve anatomical reattachment rates in patients who experience RRDCD."} +{"text": "Aspergillus species. Here we take it one step further and show that our system can be used also in a phylogenetically distinct and largely unexplored species from the genus of Talaromyces. Specifically, we exploit CRISPR-Cas9-based genome editing to identify a new gene in T. atroroseus responsible for production of polyketide-nonribosomal peptide hybrid products, hence, linking fungal secondary metabolites to their genetic origin in a species where no genetic engineering has previously been performed.The full potential of fungal secondary metabolism has until recently been impeded by the lack of universal genetic tools for most species. However, the emergence of several CRISPR-Cas9-based genome editing systems adapted for several genera of filamentous fungi have now opened the doors for future efforts in discovery of novel natural products and elucidation and engineering of their biosynthetic pathways in fungi where no genetic tools are in place. So far, most studies have focused on demonstrating the performance of CRISPR-Cas9 in various fungal model species, and recently we presented a versatile CRISPR-Cas9 system that can be successfully applied in several diverse Extracted ion chromatograms were used to evaluate the production of ZG-1494\u03b1 and talaroconvolutin A.Validated edsgaard with theS1 Appendix(DOCX)Click here for additional data file.S2 Appendix(DOCX)Click here for additional data file.S1 Fig(DOCX)Click here for additional data file.S2 FigSecond independent trial.(DOCX)Click here for additional data file.S3 Fig(DOCX)Click here for additional data file.S4 Fig(DOCX)Click here for additional data file.S5 Fig(DOCX)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file."} +{"text": "Current modifications required for metabolic cart measures of oxygen consumption in the MRI environment present clinical challenges in widespread application. Using the Fick principle we sought to quantify and compare non-invasive MRI derived Chronic heart failure (CHF) is the leading hospital discharge diagnosis in patients over age 65 [1]. Emerging techniques in cardiovascular magnetic resonance (CMR) have resulted in unique opportunity for improvement of non-invasive assessment of the physiologic and anatomic effects of CHF. We have previously demonstrated the accuracy and feasibility of 2 (ml O2/100ml blood) from quantitative T2 mapping of the left and right ventricle blood pools. The Fick principle was used to quantify 2 for comparison with measured VO2. Lin's Concordance Correlation Coefficient (\u03c1c) and the 95 percent lower confidence level (95% LCL) were used to evaluate agreement between two techniques MRI-Eight subjects underwent CMR exam with in-magnet c was 0.9669, while the 95% LCL was 0.8815.Mean 2 saturation. The combination of VO2 and CMR will potentially enhance non-invasive estimation of oxygen extraction, and when combined with exercise stress may be helpful in determining whether cardiac function or peripheral oxygen extraction, are the primary cause of heart failure related symptoms in individual patients.Non-invasive MRI derived"} +{"text": "Background: The neurobiology of the frontal network syndrome (FNS) that may occur with isolated subtentorial stroke is unknown.Aim: Evaluate for frontal network syndromes in young people post subtentorial stroke who have recovered neurologically and compare to a stroke lesion group least likely to manifest frontal network syndromes.Methods: Young people (18\u201349 years) with isolated cerebellar or brainstem subtentorial stroke (ST) that had recovered to independency (Rankin score \u08d8 2) with minimal or no residual neurological deficit (NIHSS \u08d8 4) with neurological recovery enabling resumption of former employment. Comparison was made to age and education matched young people with posterior circulation territory parieto occipital lobe infarcts (PO). Depression, anxiety, systemic disease, autoimmune disease, neurodegenerative disease and substance abuse were specific exclusions. A battery of frontal tests surveying the principal frontal network syndromes was used. Neurological deficit and long tract signs were measured by the NIH stroke score (NIHSS).Results: From the cognitive stroke registry of young stroke patients (n = 511), analysis for isolated subtentorial infarction yielded cerebellar infarcts and brainstem infarcts . After exclusions, 16 patients were compared to 10 PO infarct patients controlled for mean age, gender and NIH stroke scores. Overall 11/16 (69%) patients in the ST and 5/10 (50%) in the PO group manifested one or more of the principal FNS syndromes. Mean T scores for apathy, disinhibition, executive function and emotional intelligence standard scores were significantly more impaired in the ST group, but not for WCST error percentage scores.Conclusions: The mismatch of scant neurological deficit manifested by low NIHSS but with FNS in the majority of isolated ST stroke and more so than with PO stroke, gives support for a state dependent or neurotransmitter perturbation. The clinical impact is that such syndromes may be amenable to neuropharmacological intervention."} +{"text": "Despite advancements in ventilator technologies, lung supportive and rescue therapies, the outcome and prognostication in acute respiratory distress syndrome (ARDS) remains incremental and ambiguous. Metabolomics is a potential insightful measure to the diagnostic approaches practiced in critical disease settings. In our study patients diagnosed with mild and moderate/severe ARDS clinically governed by hypoxemic P/F ratio between 100\u2013300 but with indistinct molecular phenotype were discriminated employing nuclear magnetic resonance (NMR) based metabolomics of mini bronchoalveolar lavage fluid (mBALF). Resulting biomarker prototype comprising six metabolites was substantiated highlighting ARDS susceptibility/recovery. Both the groups (mild and moderate/severe ARDS) showed distinct biochemical profile based on 83.3% classification by discriminant function analysis and cross validated accuracy of 91% using partial least squares discriminant analysis as major classifier. The predictive performance of narrowed down six metabolites were found analogous with chemometrics. The proposed biomarker model consisting of six metabolites proline, lysine/arginine, taurine, threonine and glutamate were found characteristic of ARDS sub-stages with aberrant metabolism observed mainly in arginine, proline metabolism, lysine synthesis and so forth correlating to diseased metabotype. Thus NMR based metabolomics has provided new insight into ARDS sub-stages and conclusively a precise biomarker model proposed, reflecting underlying metabolic dysfunction aiding prior clinical decision making. Acute respiratory distress syndrome (ARDS), despite many ventilator and therapeutic measures persists with a death toll of more than 40%,which has grabbed the attention of intensivists worldwide. With a The basis of further exploratory studies in ARDS relies on omics approaches by using biofluids to monitor the cellular metabolism in the diseased state. Though genomics and protThe severity classification as per the new Berlin definition is based on the clinical index mainly P/F ratio which rules out some key ancillary variables of high risk profile thereby stressing more reliable and unambiguous insightful studies. Our study aims to better interpret ARDS severity classification using NMR based metabolomics and thereby set up metabolite credentials for a biomarker model aiding ARDS disease management and follow up. Our present study gives the distinct biochemical profile of mild and moderate/severe ARDS group based on endogenous metabolic profile. Differential biomarker candidates were categorized based on chemometric and pattern recognition methods from lung alveoli using NMR spectroscopy of mBALF. Comprehensive univariate and multivariate analysis resulted in the pivotal role of metabolites such as taurine, threonine, glutamate, proline, and lysine/arginine to metabolic perturbations in acute and mild stages of ARDS. Substantial role of these metabolites in ARDS intricate metabolic network was found to be governed by the dysregulated arginine and proline metabolism, lysine synthesis and degradation, taurine and hypotaurine metabolism, glycine, serine and threonine metabolism and glutamine and glutamate metabolism. The above mentioned analysis led to a putative biomarker model predictive of ARDS susceptibility or recuperation and the underlying dynamic biochemical behavior. Potential and prospects of the proposed biomarker model comprising these six metabolites is also evident from receiver operating characteristic (ROC) curve and significant pathway analysis impact greater than equal to 0.1. Thus metabolomics studies can correlate clinical endpoints with diseased specific markers that will be predictive, preventive and participatory. Correspondingly such biomarker model validated in large sample size holds application for early identification of high risk individuals providing metabolic and clinical variables of ARDS progression left unmapped.2O) and respiratory system compliance. Similar diagnostic measures were followed for mild patients except the P/F ratio was taken between 200\u2013300. Lung protective ventilation as per the ARDSNet protocol was practiced in the ICU. Exclusion criteria in the current study include patients with age less than 18 years, pregnancy, chronic obstructive pulmonary disease (COPD) patients, bronchial asthma, interstitial lung disease, and other chronic respiratory ailments.Total 36 samples were included in the study with 23 samples from patients diagnosed with moderate/severe ARDS and 13 samples taken from mild ARDS patients enrolled at Intensive Care Unit (ICU) admission of Sanjay Gandhi Post Graduate Institute of Medical Sciences (SGPGIMS). Patients included in the study group were suffering from mild and moderate/severe ARDS as stipulated by Berlin definition convened in the year 2012. The diagnostic criteria for moderate/severe ARDS was baseThe cross-sectional study was performed in ICU of a tertiary care medical center in SGPGIMS, Lucknow, and Centre of Biomedical Research (CBMR), Lucknow, India. The experimental protocol was approved by the SGPGIMS ethical committee and written informed consent was obtained from all subjects or their surrogate decision makers. The mBALF sample from mechanically ventilated patients was collected using the standardized non bronchoscopic \u201ccatheter in catheter\u201d technique in ICU w1H NMR mBALF spectra of mild and moderate/severe ARDS group in 800MHz NMR spectrometer (BrukerBiospin). The process of NMR spectral acquisition begins with preparation of 550\u03bcl sample including 200\u03bcl buffer to minimize the variation in pH with Trimethylsilylpropanoic acid (TSP), D2O and 350\u03bcl BALF sample. TSP (6.53mM) was added in the buffer for internal chemical shift reference along with 10% D2O to provide a field frequency lockThe samples were vortexed and then centrifuged at 16000 rpm for 10min at 4\u00b0C to remove cellular debris, bacteria and then supernatant preserved at -80\u00b0C inside freezer for storage till NMR experiments were performed. The acquisition parameters were optimized for 1H, 13C, 15N, and 2H lock) probe at 300.15K for one dimensional (1D) proton (1H) NMR data acquisition of mBALF samples. All 1D NMR spectra was based on water pre saturation pulse sequence and acquired using a 90\u00b0 flip angle, a 20ppm spectral width and a relaxation delay of 5 seconds, 32 transients were collected into 64k data points with an acquisition time (Taq) of 1.99 seconds and 16 dummy scans. 1H 1D spectra were referenced to the TSP signal (\u03b4 = 0.00 ppm). The Free induction decays (FIDs) were multiplied by an exponential weighting function corresponding to a line broadening function of 0.3 Hz and zero filled before Fourier transformation. The acquired spectra were phased and baseline corrected and manually integrated with respect to TSP for chemical shift calibration and calculating relative resonance intensities of small molecular weight metabolites. Small metabolite resonances present in 1Dspectra were assigned and confirmed by using two dimensional (2D) NMR spectra including HSQC with respect to the concentration of TSP with standard error shown as error bars and PLS-DA Component 1 = 20.8% and Component 2 = 26% with 83.3% correct classification suggesting their predominance in ARDS were further supported by various feature selection tools. The results were confirmed with volcano plot, EBAM NMR based metabolomics can differentiate mild and moderate/severe ARDS group on the basis of discrete metabolic signature distinct from the clinical and other ancillary variables. (2) This is the first differential study executed on such sample size of ARDS employing the New Berlin definition. (3) The results have been validated independently in multivariate datasets. (4)The six selective pool of endogenous markers constituting the predictive biomarker model can reflect the metabolic anomaly in ARDS substages with 95 percent accuracy defined by AUROC and the significantly associated metabolic pathway.Lung injury due to its many etiologies results in complex metabolic perturbation, consequently metabolites linked to the diseased state shows a surge of metabolites interlinked with the dysregulation of metabolism that can be used to address the various clinical manifestations. ARDS pathophysiology governing inflammatory reactions results in hypoxia, reactive oxygen species (ROS) production and oxidative stress which trigger oxidative attack on lysine, arginine, proline, threonine leading to carbonylated proteins. Carbonylated proteins which are oxidated proteins have found to be implicated in ARDS similar tMacrophage and neutrophil sequestration results in the collateral release of ROS, proinflammatory cytokines hampering epithelial and endothelial functions with increased microvascular permeability due to endothelial barrier disruption. The consequent free radical production during the pathogenesis of ARDS is likely reduced by the high concentration of taurine. Taurine is an endogenous sulphur containing amino acid found in neutrophil. Taurine has already been attributed for its protective role as an antioxidant with anti-inflammatory properties. Taurine has also been found imperative in alleviating IL-2 induced lung injury by attenuating neutrophil- endothelial interaction. Thus taAnother metabolite found proliferated in ARDS patients in our study was arginine. The key role of arginine has been signified in the intermediary metabolism of critically ill patients with salient effect in the periods of hypermetabolic stress. ArgininAnother metabolite upregulated in the diseased group is threonine. Threonine a gluconeogenic amino acid is found to be associated with sepsis which acThese metabolites can be used as a possible measure to illustrate the complex pathophysiology associated with lung injury and thus exemplifying the different metabolic pattern and its associated biochemical alterations exhibited in mild and moderate/severe ARDS. The above pilot study helped in identifying diagnostic signatures based on ARDS substages. But the subsequent biomarker credentials based on severity and progression requires more such follow up and outcome studies with validation in large sample size. The above mentioned limitation is on the pipeline to further establish the role of the proposed biomarker model in the clinical setting.In conclusion, NMR based metabolomics study was performed to fingerprint the metabolites depicting different stages of ARDS and their impact in the associated biological pathway suggesting their possible role in susceptibility/recuperation. Iterative statistics led to the compilation of 6 key metabolites which could differentiate mild and moderate/severe ARDS, thus arriving at a conclusive predictive biomarker model. The pivotal biological roles of these biomarker candidates investigated through pathway analysis can further establish their contribution to ARDS complex heterogeneity and disease manifested systemic response. The prior research findings are based on pilot study which necessitates streamlining the putative biomarker model in future prospective cohort studies to establish its diagnostic accuracy and potency. Further research into these biomarkers can give better understanding of the pathophysiology, progression and their possible utility and applicability as therapeutic targets. Our study infers NMR based metabolomics as a plausible tool with multidisciplinary applications to provide biomarker model having potent implication in ARDS management with distinct clinical relevance and downstream effect of metabolome in ARDS stages.S1 TextFigure A): Representative 800 MHz 1H\u221213C HSQC spectrum of mBALF collected from ARDS patient depicting diseased lung-specific metabolites. : Representative 800 MHz 1H\u22121H TOCSY spectrum of mBALF collected from ARDS patient depicting diseased lung-specific metabolites. : Data normalization by Pareto scaling and log transformation. Branched chain amino acids = BCA. : a) Two-dimensional and b) Three-dimensional score plot of principal component analysis with red color representing Mild ARDS and green as Moderate/ Severe ARDS, c) Two-dimensional and d) Three-dimensional score plot of partial least squares discriminant analysis with red color representing Mild ARDS and green as Moderate/ Severe ARDS e) values of the classification performance assessed by accuracy, R2 and Q2f) third component best classifies the model shown with asterisk. Principal component = PC, partial least squares discriminant analysis = PLS-DA. : Variables importance in projection (VIP). : Mean \u00b1 standard error of the nine metabolites is shown with respect to the Trimethylsilylpropanoic acid concentration (relative concentration in arbitrary unit). Trimethylsilylpropanoic acid = TSP, arbitrary unit = au, Branched chain amino acids = BCA. : a) Volcano plot with red dot showing important metabolites. b) Significant values obtained from volcano plot. PRO = Proline, LYS/ARG = Lysine/arginine, TAU = Taurine, THR = Threonine c) statistical tool empirical Bayesian analysis of metabolites to show the discerning markers d) values obtained from empirical Bayesian analysis of metabolites. Fold change = FC, false discovery rate = FDR, empirical Bayesian analysis of metabolites = EBAM. : a) and c) Random forest classification error with accuracy b) significant metabolites on the basis of mean decrease accuracy and d) Values of mean decrease accuracy. out of bag error = OOB error. (Table A): Classification results obtained from discriminant function analysis of 17 metabolites with prediction accuracy of 94.4%. (Table B): Significant metabolites selected by T-test with a threshold p value of <0.05 analogous with Variable importance in projection values. (Table C): Discriminant function analyses of nine metabolites with 88.9% correct classification. (Table D): The classification result of stepwise discriminant function analysis to weed out 5 discriminating markers with 83.3%correct classification. (Table E): Partial least squares discriminant analysis cross validation details of 6 putative biomarkers with values of the classification performance assessed by accuracy, R2 and Q2and third component best classifies the model.((PDF)Click here for additional data file."} +{"text": "TP53, PIK3CA, NOTCH1, TP63 and CDKN2A were the most frequently mutated genes. Cases were characterized by a low copy number burden with recurrent focal amplification in 11q13.3 and deletion in 15q22. Cases with low SCNAs showed an improved overall survival. We found significant correlations with decreased overall survival between focal amplified regions 4p16, 10q22 and 22q11, and losses in 12p12, 15q14 and 15q22. The mutational landscape in our cases showed an association to both environmental exposures and clinical characteristics. We confirmed that somatic copy number alterations are an important predictor of HNSCC overall survival.We investigated how somatic changes in HNSCC interact with environmental and host risk factors and whether they influence the risk of HNSCC occurrence and outcome. 180-paired samples diagnosed as HNSCC in two high incidence regions of Europe and South America underwent targeted sequencing (14 genes) and evaluation of copy number alterations (SCNAs). Head and neck squamous cell carcinomas (HNSCC) constitute a heterogeneous group of cancers, which include cancers arising at the oral cavity, nasopharynx, oropharynx, hypopharynx, and larynx. Collectively, these cancers are the seventh most common malignancy diagnosed worldwide , with arTP53, whole genome duplications and multiple recurrent chromosomal gains and losses associated to increased genomic disruption affecting cell cycle checkpoints and PI3K-AKT signaling. Advanced stage HNSCC tumours have shown mutations in more than one PI3K pathway molecule: PIK3CA, PTEN and described alterations in PI3C2G [ANXA2 gene, has been previously shown to be associated with poorly differentiated tumours in advanced cases. Decreased ANXA expression has not however, been formerly shown to be an independent prognostic factor for disease-specific survival in HNSCC [FGFR3 gene. High expression levels of FGFR3 contribute of tumour initiation and early-stage progression in HNSCC[in vitro and in vivo[FGFR3 in HNSCC.Additionally, focal copy number alterations were found to be significant prognostic markers: 22q11.2 amplification and deletions in 15q22 and 12p12 regions have been associated to smoking related tumours and advanced stage. The 22q11 region contains the lung SCC and as alung SCC . Decreasn PI3C2G , 60. Morin HNSCC , 44. We in HNSCC. More imd in vivo, highligTP53 mutations[TP53 mutation type (disruptive vrs nondisruptive or EAP53 score of missense mutations) showed no association to overall survival, either. In agreement to our results, Kim and colleagues, found that patients diagnosed with oral squamous cell carcinoma of the gingivo-buccal region (GBSCC) from the Indian Team project of the International Cancer Genome Consortium (ICGC), did not showed an association between TP53 mutation status and overall survival [Most studies on HNSCC have documented a decreased overall survival associated to mutations, 37, 63.survival . Similarsurvival .One of the main limitations of our study is the reduced number of HNSCC cases with early stage tumours. It would be important to further characterize the genomic alterations in early stages of head and neck cancer cases in order to identify biomarkers for early detection and prognostic stratification especially for the high-risk groups in regions of increase incidence. In addition, our survival analysis was limited due to the lack of complete treatment information for most cases. Treatment regimens have an important association with Head and Neck cancer overall survival and should be included in future analysis specially those involving multicentre studies.In summary, we have identified HNSCC cases with low SCNAs that differentiate as a subset of head and neck cancers driven predominantly by gene mutations and focal alterations rather than chromosomal instability events and are characterized by an improved overall survival. The mutational landscape described in our series of cases showed a clear association to both environmental exposures and clinical characteristics. Further studies integrating genomic, clinical and epidemiological data, especially in high-risk populations, are necessary to better identify high-risk stratification and characterize prognosis of head and neck cancer cases.S1 FigTP53 mutations detected in the Gencapo Series. Example: TP53Asn239Asp mutation previously detected by Sanger sequencing (B) Plots of mutational calling showing an example of independent libraries sequenced from the same case.(A) Venn diagram of number of (PDF)Click here for additional data file.S2 Fig(A) Mutually exclusive alterations between the 14 genes sequenced (B) Co-occurrence of alterations . Z score based on deviation of the observed mutations compared to expected, obtained by permuting events.(PDF)Click here for additional data file.S3 FigMutation colours represent: Green: Missense Mutations; red: Truncating Mutations , black: Inframe Mutations . Circles colored with purple indicate residues that are affected by different mutation types at the same proportion.(PDF)Click here for additional data file.S4 Fig(A) Mutational frequencies of the 14 genes sequenced in 15 HPV16E6 positive cases. (B) Comparison of Significant Focal copy number losses between HPV positive and HPV negative cases. (*) Regions significantly associated with overall survival.(PDF)Click here for additional data file.S1 TableIn red regions significantly associated with overall survival and head and neck cancer related genes.(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file."} +{"text": "The aim of this work was to prepare hydroxyapatite coatings (HAp) by a sol-gel method on Ti6Al4V alloy and to study the bioactivity, biocompatibility and corrosion protection behaviour of these coatings in presence of simulated body fluids (SBFs). Thermogravimetric/Differential Thermal Analyses (TG/DTA) and X-ray Diffraction (XRD) have been applied to obtain information about the phase transformations, mass loss, identification of the phases developed, crystallite size and degree of crystallinity of the obtained HAp powders. Fourier Transformer Infrared Spectroscopy (FTIR) has been utilized for studying the functional groups of the prepared structures. The surface morphology of the resulting HAp coatings was studied by Scanning Electron Microscopy (SEM). The bioactivity was evaluated by soaking the HAp-coatings/Ti6Al4V system in Kokubo\u2019s Simulated Body Fluid (SBF) applying Inductively Coupled Plasma (ICP) spectrometry. 3--2,5-diphenyl tetrazolium bromide (MTT) and Alamar blue cell viability assays were used to study the biocompatibility. Finally, the corrosion behaviour of HAp-coatings/Ti6Al4V system was researched by means of Electrochemical Impedance Spectroscopy (EIS). The obtained results showed that the prepared powders were nanocrystalline HAp with little deviations from that present in the human bone. All the prepared HAp coatings deposited on Ti6Al4V showed well-behaved biocompatibility, good bioactivity and corrosion protection properties. The cell viability absorbance tests have shown that the leachate obtained during a period of 7 days showed a low toxicity for cultures of osteoblasts used as model, tending moreover to disappear any sign cytotoxicity over the period of 7 days defined in the experimental design. Both obtained HAp sol-gel-derived coatings showed strong adhesion at the coating/alloy interface. In terms of corrosion protection, the HAp coating/Ti6Al4V systems thermally treated in the range 600\u2013800 \u00b0C exhibited good corrosion protection preventing the ion release from the Ti6Al4V alloy to the SBF."} +{"text": "The combination of defined factors with small molecules targeting epigenetic factors is a strategy that has been shown to enhance optimal derivation of iPSCs and could be used for disease modelling, high throughput screenings and/or regenerative medicine applications. In this study, we showed that a new first-in-class reversible dual G9a/DNMT1 inhibitor compound (CM272) improves the efficiency of human cell reprogramming and iPSC generation from primary cells of healthy donors and patient samples, using both integrative and non-integrative methods. Moreover, CM272 facilitates the generation of human iPSC with only two factors allowing the removal of the most potent oncogenic factor cMYC. Furthermore, we demonstrated that mechanistically, treatment with CM272 induces heterochromatin relaxation, facilitates the engagement of OCT4 and SOX2 transcription factors to OSKM refractory binding regions that are required for iPSC establishment, and enhances mesenchymal to epithelial transition during the early phase of cell reprogramming. Thus, the use of this new G9a/DNMT reversible dual inhibitor compound may represent an interesting alternative for improving cell reprogramming and human iPSC derivation for many different applications while providing interesting insights into reprogramming mechanisms. Induced pluripotent stem cells (iPSC) can be generated by overexpression of core pluripotency factors and reprRecent studies have enabled large-scale genomic, epigenomic and proteomic profiling of cells as they acquire pluripotency through the reprogramming process \u20137, reveaSox2 and cMyc. However, their effect in human cells and the mechanistic changes in the reprograming roadmap beyond G9a and DNMTs inhibition that enable improved iPSC derivation were not clearly investigated.Gene expression changes during the reprogramming process are accompanied by removal of epigenetic roadblocks including histone and DNA methylation marks . As examin vitro and in vivo activity . CM. CM50 fothdrawal . Furtherthdrawal , indicat+ colonies 30 days after induction, we found that CM272 increased the reprogramming efficiency almost four-fold when added 48h before doxycycline administration Fig 2A). stration . The numstration . Additiostration . We furtstration . The lac+ colonies indicating that this factor is essential for iPSC generation under these conditions and inhibited cell cycle related genes 21]. Th. Th21]. We next analyzed gene expression changes induced by CM272 after OSKM induction with doxycycline. Due to the transient effect of the CM272 treatment we focused on a set of genes described to be induced early in the reprogramming process, as well as key pluripotency-associated genes 8,10]. ,10. 8,10We further investigated the epigenetic signature of the same set of selected genes significRecent studies have shown that initial binding of OSKM to particular genomic regions is essential for pluripotency induction . We furtTaken together those results suggests that CM272 treatment improves TF engagement at promoter regions of genes involved in cell reprogramming. Since CM272 is a reversible dual G9a/DNMT inhibitor we also investigate the methylation status by pyrosequencing of the MET and pluripotency promoter regions during early phase of cell reprogramming in the presence of CM272. We were not able to detect significant changes probably due to the longer CM272 treatment required to reduce DNA methylation . TogetheIn this study, we showed that a new first-in-class dual and reversible G9a/DNMT inhibitor compound improvesSox2 and reprogramming of MEFs only with Oct4 and Klf4 required the use of BIX-01294 together with RG108, a DMNT inhibitor, or BayK, an L-calcium channel agonist. As CM272 also inhibits DNMT and inhibition of DNMT improves iPSC generation [The use of small molecules to overcome reprogramming barriers, like G9a/GLP mediated H3K9 methylation ,18, reprneration it is teneration ,39. Neve50. This effect was especially dramatic with CM579, fact that led us to discard this compound, focusing our efforts on the characterization of CM272 in cell reprogramming. At doses close to GI50, CM272 reduced the expression of several cyclins and CDKs, effect that could limit the efficacy for enhanced iPSC generation since induction of cell proliferation has been described to be important during the first stages of cell reprogramming [Our reversible dual G9a/DNMT inhibitor lead compounds CM272 and CM579 impaired cell growth even at doses below GIgramming . Howevergramming reportingramming and potegramming and its gramming . Our tragramming , suggestIn this work, we have focused in the mechanisms underlying the improvement of cell reprogramming with CM272. Our results indicate that CM272 promotes higher expression of MET and pluripotency associated genes involved in cell reprogramming ,8. TheseIn summary, this study takes a closer look at the early events occurring during cell reprogramming of human cells after treatment with a G9a/DNMT reversible dual small molecule inhibitor compound. The increased understanding of the mechanism of enhanced reprogramming will provide strategies to improve its utility for applications like disease modelling. These results indicate that the reduction of repressive marks before induction of reprogramming facilitates the outcome of the process, suggesting that this strategy could be applied not only to iPSC generation, but also to other reprogramming processes, like trans-differentiation or direct reprogramming. Moreover, G9a/DNMT inhibitors could also have positive effects improving differentiation processes where chromatin status and epigenetic factors remodeling are also required.S1 Fig(A) Chemical structure of CM272 and CM579 compounds. (B) Cell cycle analysis in BJ cells treated with three different concentrations of CM272 or CM579 for 48 hours. (C and D) Dot-blot analysis of 5mC levels after treatment for 7 days (A) or 48h (B) of BJ cells with the indicated doses of CM272. (E and F) Quantification of dot-blot intensities from at least 4 independent experiments.(TIF)Click here for additional data file.S2 Fig+ colonies at day 30 of cell reprogramming in primary cells treated with CM272 (200nM). Mock indicates no CM272 treatment.(A) Representative pictures and quantification of GFP expression levels observed in doxycycline-inducible-GFP-infected BJ cells after doxycycline addition in the presence or absence of CM272. (B) Quantification of transcription factor expression levels observed in doxycycline-inducible-TF-infected BJ cells after doxycycline addition in the presence or absence of CM272. Error bars represent SD of three independent experiments. (C) Representative images of AP(TIF)Click here for additional data file.S3 Fig(A) Hierarchical cluster analysis of the microarray data of OSKM infected BJ cells after CM272 treatment just before doxycycline induction. (B) Western blot of H3K9me2 levels after CM272 treatment of the three independent experiments. (C) Venn diagram of commonly differentially expressed genes between CM272-treated cells, pluripotency-associated genes and genes involved in early events in cell reprogramming. (D) Enrichment analysis at the major dynamic expression patterns during human iPSC generation of differentially expressed genes in OSKM-infected BJ cells after CM272 treatment and before doxycycline addition. (E) Differential expression (LogFC) of enriched genes of the early reprogramming events involving early and late somatic categories in the major dynamic expression patterns during human iPSC generation .(TIF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S1 File(DOCX)Click here for additional data file."} +{"text": "In 2014\u20132016, >1,000 wild goats and sheep in 4 northern and central provinces of Iran died from peste des petits ruminants virus (PPRV) infection. Partial nucleoprotein sequencing of PPRV from 3 animals showed a close relationship to lineage 4 strains from China. Control measures are needed to preserve vulnerable ruminant populations. Morbillivirus, family Paramyxoviridae) causes a highly contagious disease with a high death rate in wild and domestic small ruminants. Four PPRV lineages (L1\u2013L4) exist in Africa and Asia and sheep , which have become extinct in several West Asia countries, are considered vulnerable species in Iran (Gazella subgutturosa) with clinical signs similar to those caused by PPRV infection died in Kavir National Park . In addition, we performed partial nucleocapsid gene sequencing of 3 PPRV isolates from 2015; results showed 100% pairwise nt identity among the isolates . The strains shared highest nt identity (99.4%) with PPRV-L4 strains that were circulating in domestic or wild small ruminants in northwestern and southeastern China during 2013\u20132015 , and location and number of dead wild goats and wild sheep and phylogenetic tree of PPRV strains, Iran, 2014\u20132016."} +{"text": "ORC1, ORC4, ORC6, CDC6, CDT1, and MCM5). These genes encode components of the pre-replication complex, which assembles at origins of replication prior to S phase. Also, variants in two additional replication initiation genes have joined the list of causative mutations for MGS (Geminin and CDC45). The identity of the causative MGS genetic variants strongly suggests that some aspect of replication is amiss in MGS patients; however, little evidence has been obtained regarding what aspect of chromosome replication is faulty. Since the site of one of the missense mutations in the human ORC4 alleles is conserved between humans and yeast, we sought to determine in what way this single amino acid change affects the process of chromosome replication, by introducing the comparable mutation into yeast (orc4Y232C). We find that yeast cells with the orc4Y232C allele have a prolonged S-phase, due to compromised replication initiation at the ribosomal DNA (rDNA) locus located on chromosome XII. The inability to initiate replication at the rDNA locus results in chromosome breakage and a severely reduced rDNA copy number in the survivors, presumably helping to ensure complete replication of chromosome XII. Although reducing rDNA copy number may help ensure complete chromosome replication, orc4Y232C cells struggle to meet the high demand for ribosomal RNA synthesis. This finding provides additional evidence linking two essential cellular pathways\u2014DNA replication and ribosome biogenesis.A form of dwarfism known as Meier-Gorlin syndrome (MGS) is caused by recessive mutations in one of six different genes repeats is severely compromised in yeast that harbor the MGS allele. Consequently, cells that have reduced their rDNA copy number from ~150 to fewer than 30 copies overtake the culture. Although the loss of rDNA repeats helps ensure the complete replication of chromosome XII during S phase, cells with fewer rDNA repeats struggle to meet the high demand for ribosomal RNA.The origin recognition complex (ORC) is essential for licensing replication origins during M/G1 for their firing in the subsequent S phase. Individuals with a rare form of dwarfism called Meier-Gorlin syndrome (MGS) have mutations in proteins required for origin activation, including various subunits of ORC. To better understand the molecular and cellular consequences of these mutations, we introduced an equivalent MGS mutation in Saccharomyces cerevisiae, where chromosomal origins were first identified by their ability to maintain recombinant plasmids after transformation into yeast . We. WeORC4 repeats . Our finORC4 has been shown to be causative for some instances of MGS in humans . Ea. Ea41]. orc4Y232C cells was such a striking phenomenon, we tested six additional isolates of the mutant to determine if this size reduction was a consistent phenotype of the orc4Y232C mutation. Immediately after selection for loss of the ORC4 allele (~20 generations), all six isolates had a smaller chromosome XII than wild type, with an rDNA locus of ~30\u2013100 repeats . Rather orc4Y232C allele. To test this hypothesis, we introduced a different MGS-like mutation into budding yeast. Some cases of MGS have been reported to be caused by mutations in CDC45, a core component of the CMG complex, the replicative helicase that travels with the replisome during S phase ..H. sapie(TIF)Click here for additional data file.S2 Fig(A).Superimposition of the Hs and Sc Orc4 subunits highlights the high degree of structural similarity between the proteins from the two species. The right panel focuses on the Tyrosine mutated in human MGS patients (Tyr174) and the corresponding Tyrosine in yeast (Tyr232) with the side chains of these amino acids displayed in red.(B).Structural models of ScOrc1-6 and ScCdc6 in complex with a double stranded DNA molecule. The right panel focuses on Tyr232 of ScOrc4, with the side chain of this amino acid depicted as red spheres.(TIF)Click here for additional data file.S3 FigORC4 (black) and orc4Y232C (red) are shown for cells after exposure to HU for 30 min. The relative ratio of ssDNA (S/G1) is plotted against chromosome coordinates (kb). A yellow circle denotes centromere locations and the positions of verified origins of replication are marked by orange triangles. The rDNA locus and adjacent flanking sequence on Chr XII (cf. 440\u2013490 kb) were omitted due to insufficient probe coverage on the microarray slide.Genome wide ssDNA profiles for (TIF)Click here for additional data file.S4 Figorc4Y232C after growth for ~100 generations. Change in Chr XII size was measured by CHEF gel electrophoresis. Left, ethidium bromide stained image; right, Southern blot image following hybridization with a Chr XII-specific single-copy sequence. By ~100 generations the size of Chr XII had stabilized at ~30 copies of rDNA for most of the population in all six isolates of orc4Y232C.Variation in rDNA copy number was analyzed in the six isolates (a-f) of (TIF)Click here for additional data file.S5 FigCDC45 due to loss of rDNA repeats.Left, ethidium bromide stained image; right, Southern blot hybridization for Chr XII. All five isolates had a smaller Chr XII than (TIF)Click here for additional data file.S6 Figorc4Y232C with ~30 copies of rDNA (lane #2) was transformed with a centromere plasmid (pRS415) containing a copy of either ORC4 or orc4Y232C. rDNA copy number was analyzed in three isolates from each transformation. An increase in rDNA copy number was observed in cells transformed with the plasmid containing ORC4; however, no increase in rDNA copy number was observed in cells transformed with the plasmid containing orc4Y232C.An isolate of (TIF)Click here for additional data file.S7 FigRed arrow indicates the polymorphism in the ACS.(TIF)Click here for additional data file.S8 FigORC4 rDNARM and orc4Y232CrDNARM cells generated by measuring the optical density over time of mid-log cultures in synthetic complete medium at 30\u00b0C. The mutant (white circle) shows a substantial growth defect with a doubling-time 54 minutes (27%) longer than wild-type cells (black circle).Growth curves of (TIF)Click here for additional data file.S9 FigORC4 rDNARM (black) and orc4Y232CrDNARM (red) are shown for cells after exposure to HU for 30 min. The relative ratio of ssDNA (S/G1) is plotted against chromosome coordinates (kb). A yellow circle denotes centromere locations and the positions of verified origins of replication are marked by orange triangles. The rDNA locus and adjacent flanking sequence on Chr XII (cf. 440\u2013490 kb) were omitted due to insufficient probe coverage on the microarray slide.Genome wide ssDNA profiles for (TIF)Click here for additional data file.S10 FigORC4 rDNABY, ORC4 rDNARM, orc4Y232CrDNABY, and orc4Y232CrDNARM) were measured and pair-wise comparisons of those values at each origin between the different strains are shown in the three scatter plots.The relative areas under the peaks for the four strains ((TIF)Click here for additional data file.S11 FigAn ethidium bromide stained gel image of the total nucleic acid content from exponentially growing cells separated by electrophoresis. The blue dashed line indicates where the gel was cut so that the two different parts of the gel containing either genomic DNA or rRNA could be separately treated for Southern or northern transfer to hybridization membranes. The hybridization images and quantifications are shown in (TIF)Click here for additional data file.S12 FigORC4 rDNABY (blue) and orc4Y232CrDNABY (red) cells harboring a GFP tagged version of the single-copy ribosomal protein Rpl10. Cells were grown to mid-log phase and relative fluorescence was measured by flow cytometry.Relative fluorescence of (TIF)Click here for additional data file.S13 FigORC4 (blue circles) and orc4Y232C (red circles) specific origins are shown across the sixteen yeast chromosomes aligned by their centromeres at x = 0. Centromere locations are marked by a yellow circle. (B) The locations of only the ORC4 specific origins are shown relative to the locations of centromeres (yellow line). (C) The locations of only the orc4Y232C specific origins are shown relative to the locations of centromeres (yellow line).(A) The locations of (TIF)Click here for additional data file.S1 Table(PDF)Click here for additional data file.S1 File(PY)Click here for additional data file."} +{"text": "Cucumis sativus with the movement protein (MP) of Cucumber mosaic virus (CMV). This was identified first in a yeast two-hybrid screen and validated by in vivo pull down and bimolecular fluorescence complementation (BiFC) assays. The BiFC assay showed localization of the bimolecular complexes of these proteins around the cell wall periphery as punctate spots. The expression of CsAO4 was induced during the initial infection period (up to 72 h) in CMV infected Nicotiana benthamiana plants. To functionally validate its role in viral spread, we analyzed the virus accumulation in CsAO4 overexpressing Arabidopsis thaliana and transiently silenced N. benthamiana plants (through a Tobacco rattle virus vector). Overexpression had no evident effect on virus accumulation in upper non-inoculated leaves of transgenic lines in comparison to WT plants at 7 days post inoculation (dpi). However, knockdown resulted in reduced CMV accumulation in systemic (non-inoculated) leaves of Nb\u0394AO-pTRV2 silenced plants as compared to TRV inoculated control plants at 5 dpi (up to 1.3 fold difference). In addition, functional validation supported the importance of AO in plant development. These findings suggest that AO and viral MP interaction helps in early viral movement; however, it had no major effect on viral accumulation after 7 dpi. This study suggests that initial induction of expression of AO on virus infection and its association with viral MP helps both towards targeting of the MP to the apoplast and disrupting formation of functional AO dimers for spread of virus to nearby cells, reducing the redox defense of the plant during initial stages of infection.Plant viral movement proteins facilitate virion movement mainly through interaction with a number of factors from the host. We report the association of a cell wall localized ascorbate oxidase (CsAO4) from To establish successful infection in a susceptible host, viruses initially move from cell to cell followed by systemic movement through phloem sieve elements. Viral movement proteins (MPs) play a dynamic role in the spread of viruses by managing their transport through symplasmic connections i.e. plasmodesmata (PD). MPs mediate viral spread either in the form of virions or as viral ribonucleoprotein (vRNP) complexes (reviewed in \u20133).Potexvirus have their MPs organized as a triple gene block (TGB) and are speculated to target PD by endoplasmic reticulum (ER) association , (a previously characterized subgroup II CMV isolated in our lab from cucumber) and clonC. sativus plants cv. Summer Green (SG). The virus was transferred through mechanical inoculation of pure culture of CMV-SG, which was maintained on N. tabacum cv. Samsun, and all the plants were kept in insect proof glasshouse conditions at 24\u00b12\u00b0C. Leaf samples were collected at different time intervals: 1, 6, 12, 24, 36, 48 and 72 h at the 2\u20133 leaf stage and from these samples Poly (A)+ RNA (~500 ng) was isolated using the mTRAP kit . The RNA was used for cDNA synthesis using SMART\u00ae technology of the Matchmaker library construction and screening kit . This was followed by generation of double-stranded cDNA utilizing the high fidelity 50\u00d7Advantage 2 polymerase mix . A GAL4 AD fusion library was produced by the co-transformation method in AH109. The screening was done on minimal selection media plates SD/-LTH or SD/-LTHA along with positive (pGADT7-RecT+ pGBKT7-53) and negative controls (pGADT7-RecT+ pGBKT7-Lam) for 7\u201310 days at 30\u00b0C. Further confirmation of positive interactions was carried out on SD/-LTHA containing 4mM AT and by \u03b2-X-gal assay.For cDNA library preparation, total RNA was extracted using the guanidine isothiocyanate method from thehttp://cucumber.genomics.org.cn/). The partial sequences obtained from the library were taken as references to design primers for the full-length genes.Positive transformants were analyzed by colony PCR. The amplified products were sequenced and data was analyzed through Blast analysis of the cucumber genome database . Percent identity index of AO from different plants (the nucleotide sequences were taken from the NCBI database) was carried out using Bioedit version 7.1.9 software , Prunus (KC152937), Tomato (AY971876), and Maize (EU973211).(A) Phylogenetic analysis of cucumber (TIF)Click here for additional data file.S3 FigThe figure showing strength of interaction of full length AO and its domains with MP domains as indicated by growth on selection plate along with positive and negative controls.(TIF)Click here for additional data file.S4 FigBased on the results of LigPlot+ interaction plot was generated showing hydrogen and hydrophobic interactions between residues. Chain A and Chain B denotes CsAO4 and CMV MP respectively.(TIF)Click here for additional data file.S5 FigEarly flowering observed in AO-overexpressing transgenic lines in comparison to wild type plants. Representative pictures of T2 plants and WT were shown.(TIF)Click here for additional data file.S6 FigAnthocyanin pigmentation was observed in AO-overexpressing transgenic lines in comparison to wild type plants. Representative pictures of upper and lower leaves of CMV infected transgenic and WT plants after two weeks were shown in the figure.(TIF)Click here for additional data file.S7 Fig\u0394AO) and CsAO4 (CsFAO and Cs\u0394AO) indicated by blue bars and cupredoxin domain 1 (CuRO1), domain 2 (CuRO2) and domain 3 (CuRO3) indicated by solid black bars.The figure showed regions of NbAO (Nb(TIF)Click here for additional data file.S8 FigCsAO4-pTRV2 (CsFAO-pTRV2): developed chlorotic patches along with necrotic areas above infiltrated leaves; (C) Partial CsAO4-pTRV2 (Cs\u0394AO-pTRV2): caused symptoms like leaf deformation, necrotic areas, and size reduction in new emerging leaves.Phenotypic effect of NbAO knockdown was observed on plants. (A-B) Full length (TIF)Click here for additional data file.S9 FigpCAMBIA1302 constructs were bombarded on onion epidermal peels by biolisitc method and GFP fluorescence was observed under fluorescent microscope. (A) Empty pCAMBIA1302 vector showing free GFP localization in cytoplasm and nucleus. (B) CMV MP-pCAMBIA1302 and (C) CsAO4-pCAMBIA1302 showing localization around cell wall region.(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file."} +{"text": "Vigna mungo) is one of primary legumes cultivated throughout India, Cv.T9 being one of its common high yielding cultivar. This article reports RNA sequencing data and a pipeline for prediction of novel long non-coding RNAs from the sequenced data. The raw data generated during sequencing are available at Sequence Read Archive (SRA) of NCBI with accession number- SRX1558530Blackgram ( Specifications TableValue of the data\u2022Vigna mungo.This is the first report of long non-coding RNAs in \u2022Vigna mungo.This study will enable researchers to identify lncRNAs of interest in a high protein yielding legume, \u2022Vigna mungo an in depth analysis with some adjustments which may pave the way for identification of lncRNAs in other non model plants as well.This article also contains a pipeline for identification of long non-coding RNAs in 1Vigna mungo (Blackgram) Cv. T9. This cultivar is widely cultivated in different states of India due to high agronomic yield; however, it is highly susceptible to Mungbean Yellow Mosaic India Virus (MYMIV) infection mediated by the vector whitefly (Bemisia tabaci).This works reports the long non-coding RNAs identified in common Indian cultivar of 22.1SRX1558530.Sample preparation for RNA isolation was done as described by Kundu et al. 2.2de-novo assembly of transcripts using Trinity De novo transcript statistics are provided in Glycine max (65%) . Remaining 2874 (The pipeline shown in ax (65%) A. Unannoing 2874 reads ar2.2.1Simple sequence repeats were predicted using MISA-MIcroSAtellite identification tool"} +{"text": "Xanthomonas oryzae pv. oryzae (Xoo), is one of the most devastating rice diseases worldwide. The development and use of disease-resistant cultivars have been the most effective strategy to control bacterial blight. Identifying the genes mediating bacterial blight resistance is a prerequisite for breeding cultivars with broad-spectrum and durable resistance. We herein describe a genome-wide association study involving 172 diverse Oryza sativa ssp. indica accessions to identify loci influencing the resistance to representative strains of six Xoo races. Twelve resistance loci containing 121 significantly associated signals were identified using 317,894 single nucleotide polymorphisms, which explained 13.3\u201359.9% of the variability in lesion length caused by Xoo races P1, P6, and P9a. Two hotspot regions (L11 and L12) were located within or nearby two cloned R genes (xa25 and Xa26) and one fine-mapped R gene (Xa4). Our results confirmed the relatively high resolution of genome-wide association studies. Moreover, we detected novel significant associations on chromosomes 2, 3, and 6\u201310. Haplotype analyses of xa25, the Xa26 paralog , and a Xa4 candidate gene (LOC_11g46870) revealed differences in bacterial blight resistance among indica subgroups. These differences were responsible for the observed variations in lesion lengths resulting from infections by Xoo races P1 and P9a. Our findings may be relevant for future studies involving bacterial blight resistance gene cloning, and provide insights into the genetic basis for bacterial blight resistance in indica rice, which may be useful for knowledge-based crop improvement.Bacterial blight, which is caused by Xanthomonas oryzae pv. oryzae (Xoo), is one of the most devastating diseases of cultivated rice (Oryza sativa L.) in tropical and temperate regions worldwide were significantly associated with resistance to P9a and P1, respectively. These results provide valuable information for future studies involving bacterial blight resistance gene cloning. Moreover, the identified diverse rice accessions carrying fine-mapped and novel loci from nearly 20 countries will provide more available donors in breeding programs aiming at developing bacterial blight resistance in different rice growing regions.We identified 12 bacterial blight resistance loci containing 121 significantly associated signals using 317,894 SNPs. Two hotspot regions were located within or nearby cloned Our study provides new insights into the genetic basis of the evolution of bacterial blight resistance in rice. The findings reported herein may be useful for knowledge-based crop improvement. Future research will focus on validating the effects of these candidate genes and their functional variants. We will use genetic transformations and DNA insertion mutant screens to verify that these genes confer bacterial blight resistance to rice.S1 Fig(A) Neighbor-joining tree of 172 accessions. (B) Principal component analysis plots for the first two components of 172 accessions. (C) Distribution of the estimated subpopulation components for each accession as determined by ADMIXTURE.(PDF)Click here for additional data file.S2 FigXanthomonas oryzae pv. oryzae (Xoo) races is indicated as the negative logarithm of the p value for the linear mixed effects model. P1 (strain PXO61). P6 (strain PXO99). C5 (strain GD1358). P3c (strain PXO340). P9a (strain PXO339). GV (strain V). The strength of the associations for the lesion lengths caused by representative strains of six (PDF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file."} +{"text": "Aortic valve disease is a complex process characterized by valve interstitial cell activation, disruption of the extracellular matrix culminating in valve mineralization occurring over many years. We explored the function of the retinoblastoma protein (pRb) in aortic valve disease, given its critical role in mesenchymal cell differentiation including bone development and mineralization.RB1 alleles. Aged pRb cKO animals showed significantly more aortic valve regurgitation by echocardiography compared to pRb het control animals. The pRb cKO aortic valves had increased leaflet thickness without increased cellular proliferation. Histologic studies demonstrated intense \u03b1-SMA expression in pRb cKO leaflets associated with disorganized extracellular matrix and increased leaflet stiffness. The pRb cKO mice also showed increased circulating cytokine levels.We generated a mouse model of conditional pRb knockout (cKO) in the aortic valve regulated by Tie2-Cre-mediated excision of floxed Our studies demonstrate that pRb loss in the Tie2-lineage that includes aortic valve interstitial cells is sufficient to cause age-dependent aortic valve dysfunction. Aortic valve disease is a complex disease process characterized by progressive thickening and fibrosis of the aortic valve (AoV) leaflets leading to valve sclerosis, a precursor to valve mineralization and often frank bone formation that can cause restriction and/or regurgitation of blood flow from the heart. Statistics from 2017 indicate that more than 100,000 patients received an aortic valve replacement as medicDuring embryonic heart development, cells derived from the endocardium , the secVICs produce an array of precisely oriented structural matrix proteins necessary for the leaflet to bear the load during diastole, while also sufficiently flexible to impart negligible outflow resistance during systole , 16. In The retinoblastoma protein (pRb) regulates two key factors relevant to aortic valve disease: maintenance of mesenchymal cell differentiation as well as bone formation and soft tissue calcification \u201327. GiveRB1fl/fl) mice and B) Von Kossa stains demonstrate the lack of calcification in pRb het or cKO animals.(TIFF)Click here for additional data file.S3 FigA) Masson\u2019s Trichrome, B) Movat pentachrome, and C) \u03b1-SMA Immunohistochemistry demonstrating the lack of distinct alteration in leaflets of 2 month old mice without pRb. Scale bar is 50\u03bcm.(TIFF)Click here for additional data file.S4 FigNormalized spectral counts of structural ECM proteins collected through LC-MS/MS analysis of pRb het (blue) and pRb cKO (red) aortic valve leaflets (n = 3 for both conditions). Mean \u00b1 Std Dev; P-value (*p<0.05) from student\u2019s t-test, corrected for multiple measures by Holm-Sidak method.(TIFF)Click here for additional data file.S5 FigA) Representative photo of spleens harvested from aged pRb het and cKO mice. B) Spleen weight normalized to tibia length, showing significantly splenomegaly in cKO mice . C) H&E staining of spleens from aged mice, showing increased white pulp in cKO samples as well as loss of typical nodular tissue structure, as in het spleens. D) H&E staining of the distal end of the tibia of aged mice, showing increased BM cells and reduced adipocytes in cKO animals. E) Representative FACS analysis of bone marrow, demonstrating increase of monocytes in cKO mice, as determined by cell surface expression Ly6C.(TIFF)Click here for additional data file."} +{"text": "Ebstein's anomaly (EA) has variable prognosis. New MRI technology allowing the measurement of vessel blood flow and oxygen content could provide additional prognostic information in the setting of fetal EA.We measured fetal weight, brain weight and lung volume in normal and EA fetuses using MRI. Blood flow and T2 in the major fetal vessels were measured using our previously published technique. Fetal oxygen delivery (DO2), consumption (VO2) and extraction fraction (OEF) were calculated using estimated fetal hemoglobin concentration.We studied 30 normal and 8 EA fetuses at 36 weeks gestation. There were 2 deaths in the 4 patients that underwent surgery . We found reduced flows in all measured vessels except ascending aorta Table . There wMRI revealed a ~50% reduction in CVO in EA fetuses compared to normal fetuses, which is greater than any other type of CHD. While EA subjects had similar placental function (indicated by similar UV T2), lower CVO resulted in reduced UV flow, therefore decreased DO2. EA fetuses had increased OEF compensating for the reduced DO2, but body and brain development were still reduced. In addition, MRI-based lung volume measurements, UV flow and VO2 maybe associated with postnatal outcome in EA fetuses. Therefore, with more experience, the MRI technique may provide useful prognostic information in Ebstein's anomaly pregnancies."} +{"text": "Dose-ranging studies were performed to identify the optimal antigen dosages based on systemic and mucosal immune responses in guinea pigs and determine any antigenic interference. A dose-dependent increase in systemic and mucosal immunogenicity against each of the VLPs were observed as well as a boosting effect for each VLP after the second dosing. A total antigen dose of \u226550 \u03bcg of each GI and GII.4 VLPs was determined to be the maximally immunogenic dose in guinea pigs. The immunogenicity results of this bivalent formulation, taken together with previous work on monovalent GelVac\u2122 norovirus vaccine formulation, provides a basis for future development of this norovirus VLP vaccine.The global health community is beginning to understand the burden of norovirus-associated disease, which has a significant impact in both developed and developing countries. Norovirus virus like particle (VLP)-based vaccines are currently under development and have been shown to elicit systemic and mucosal immune responses when delivered intranasally. In the present study, we describe the use of a dry powder formulation (GelVac Norovirus are responsible for over 90% of all non-bacterial gastroenteritis epidemics and are a leading cause of global diarrhea . La. La14]. \u2122 vaccine formulations containing GI and GII.4 norovirus VLPs. Animals were dosed on days 0 and 21 with varying amounts of the bivalent norovirus GI and GII.4 VLPs and 68% (p = 0.068) against moderate to severe disease. These vaccines were formulated with Aluminum Hydroxide and Monophosphoryl lipid adjuvants. These vaccines are still in clinical development. A norovirus-rotavirus protein combination vaccine is in preclinical development with promising results [\u2122 is made from materials that the FDA has considered generally regarded as safe (GRAS). The data presented herein, as well as previous work [\u2122 dry powder norovirus vaccine include a comparison of IN and IM administration.There are currently no licensed vaccines for norovirus. There are many factors that complicate the development of norovirus vaccines including the lack of appropriate model systems to explore vaccine target efficacy, unknown duration of protective immunity, antigenic variation and drift within genogroups and genotypes, and the unknown effects of pre-exposure history. The recent demonstration of the cultivation of norovirus in cell culture should permit human host pathogen interactions and allow assessment of various methods to prevent and treat norovirus infections . Despite results . In addi results . One oth results . The stuous work , 16, dem\u2122 dry powder bivalent norovirus vaccine extends the results from previous studies [\u2122 nasal powder on the production of systemic and mucosal anti-norovirus specific antibodies. A dose content of 15\u201350 \u03bcg GI or GII.4 VLP antigen appeared to be required to produce a robust response. Future studies will be conducted with GelVac\u2122 GI and GII.4 bivalent vaccine formulation administered either intranasally or intramuscularly. The results will help to demonstrate the efficacy of the intranasal formulation when compared to an intramuscular injection and determine the antigen dose for this bivalent vaccine that could be used in additional pre-clinical development.The data presented in this study using the GelVac studies , 16. TheS1 FigVLPs were diluted in 4x SYPRO orange solution and the melt curve was analyzed using a fluorescent thermocycler. Data is plotted as the change in Fluorescence per unit Temperature. A. Norovirus GI VLP melt curve. B. Norovirus GII.4 VLP melt curve.(TIF)Click here for additional data file.S2 FigVLP samples (0.2 \u03bcg/mL) were treated at varying temperatures for 5 minutes. Each sample was then analyzed by capture ELISA.(TIF)Click here for additional data file.S3 Fig(A) (SDS-PAGE), (B) GI VLP (western blot), and (C) GII.4 VLP (western blot).(TIF)Click here for additional data file.S4 FigSerum samples were analyzed for norovirus-specific IgG1 antibodies against GI (A) and GII.4 (B), and norovirus-specific IgG2 antibodies against GI (C) and GII.4 (D). Horizontal dotted line depicts the limit of detection for these assays.(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.S4 Table(DOCX)Click here for additional data file.S5 Table(DOCX)Click here for additional data file."} +{"text": "Imidacloprid is a neonicotinoid pesticide heavily used by the agricultural industry and shown to have negative impacts on honey bees above certain concentrations. We evaluated the effects of different imidacloprid concentrations in sugar syrup using cage and field studies, and across different environments. Honey bee colonies fed sublethal concentrations of imidicloprid over 6 weeks in field trials at a desert site (Arizona), a site near intensive agriculture (Arkansas) and a site with little nearby agriculture but abundant natural forage (Mississippi) were monitored with respect to colony metrics, such as adult bee and brood population sizes, as well as pesticide residues. Hive weight and internal hive temperature were monitored continuously over two trials in Arizona. Colonies fed 100 ppb imidacloprid in Arizona had significantly lower adult bee populations, brood surface areas and average frame weights, and reduced temperature control, compared to colonies in one or more of the other treatment groups, and consumption rates of those colonies were lower compared to other colonies in Arizona and Arkansas, although no differences in capped brood or average frame weight were observed among treatments in Arkansas. At the Mississippi site, also rich in alternative forage, colonies fed 5 ppb imidacloprid had less capped brood than control colonies, but contamination of control colonies was detected. In contrast, significantly higher daily hive weight variability among colonies fed 5 ppb imidacloprid in Arizona suggested greater foraging activity during a nectar flow post treatment, than any other treatment group. Imidacloprid concentrations in stored honey corresponded well with the respective syrup concentrations fed to the colonies and remained stable within the hive for at least 7 months after the end of treatment. A-D. Intercept and scale parameters, and associated distribution scale and shape parameters, for a regression-based fit of a Weibull distribution with censoring to bee survivorship in cage studies over two trials.(PDF)Click here for additional data file.S2 FileTablesA and B. Analysis and post hoc contrast results for hive inspection data from a field experiment conducted in Arizona 2014.(PDF)Click here for additional data file.S3 FileTables A and B. Analysis and post hoc contrast results for continuous weight and temperature data from a field experiment conducted in Arizona 2014.(PDF)Click here for additional data file.S4 FileTables A and B. Analysis and post hoc contrast results for hive inspection and continuous monitoring data from a field experiment conducted in Arizona 2015.(PDF)Click here for additional data file.S5 FileTables A and B. Analysis and post hoc contrast results from a field experiment conducted in Mississippi 2015.(PDF)Click here for additional data file.S6 File(XLSX)Click here for additional data file.S1 Table(PDF)Click here for additional data file.S2 Table(PDF)Click here for additional data file.S3 Table(PDF)Click here for additional data file.S4 Table(PDF)Click here for additional data file."} +{"text": "Scientific Reports 10.1038/s41598-017-14781-3, published online 31 October 2017Correction to: The Authors missed out a previous study on a similar topic. The additional reference is listed below as reference 1, and should appear in the text as below.In the Discussion section,3 and TOC MARs are considered here as proxies for past productivity variations in response to changes in regional forcing like surface water hydrography and nutrient supply in the BoB.\u201d\u201cCaCOshould read:3 and TOC MARs are considered here as proxies for past productivity variations in response to changes in regional forcing like surface water hydrography and nutrient supply in the BoB. An earlier study by Phillips et al. has discussed possible influence of salinity/hydrographic changes on marine productivity indicated by CaCO3 MAR in the northern Bay of Bengal1.\u201d\u201cCaCOand:3 MAR).\u201d\u201cOverall diminished fresh water flux during colder and arid sub-stages caused thinning of low salinity cap leading to destabilization of the water column stratification which triggered enhanced nutrient entrainment by wind driven processes and convective mixing leading to enhanced productivity (enhanced CaCOshould read:3 MAR)1.\u201d\u201cOverall diminished fresh water flux during colder and arid sub-stages caused thinning of low salinity cap leading to destabilization of the water column stratification which triggered enhanced nutrient entrainment by wind driven processes and convective mixing leading to enhanced productivity (enhanced CaCO"} +{"text": "Sus scrofa) survival and death in 2 tuberculosis-endemic populations with different harvest pressure in Spain. Overall, tuberculosis accounted for 30% of total deaths. Increased survival in protected areas has direct implications for wild boar management and tuberculosis control.We investigated adult Eurasian wild boar ( Sus scrofa) population dynamics and hunting strategies might influence the persistence of disease is a reemerging multihost infectious disease caused by the In the context of growing and expanding wild boar populations and of increasing concern about the effect of wild boar infections . Do\u00f1ana National Park (DNP) is a protected area on the Atlantic coast of southern Spain. Harvest is part of population control management because no recreational hunting is allowed within the park; this modality has a minimal extraction capacity (1.11 wild boar/km2) and targets wild boar seen by park rangers (random and opportunistic). Wild boars, except piglets, have no natural predators in the study areas.We compared 2 settings: a mosaic of game estates and a protected area. Montes de Toledo (MT) is a mountain chain in the central Spanish plateau whose large game estates are mainly devoted to recreational hunting. Harvest is conducted by dog-driven hunts; average annual extraction quota is 2.26 wild boar/kmDuring 2009\u20132014, we captured Table foWe detected serum antibodies to MTC in 35 (78%) of 45 GPS-collared wild boars. We found no differences in MTC serum antibody prevalence between study sites (p>0.05 by Fisher exact test) and no differences in survival time between antibody-positive and -negative wild boars (p>0.05 by Mann-Whitney U test). MTC infection was confirmed by culture in 13 (72%) of 18 wild boars for which postmortem results were available. The 9 wild boars that died of generalized TB had severe lesions in >1 anatomic region; >70% of lung tissue was affected Figure 1We assessed total survival probability and the main causes of death . The mea2\u00a0=\u00a011.42, 1 d.f.; p = 0.001). Kaplan-Meier survival probabilities and causes of death are detailed by study area in Mean survival time was twice as long in DNP (average 672 \u00b1 96 days) as in MT contrasts with results obtained in other parts of Europe wild boars, thus reducing the profitability of the hunting industry. Hunters should therefore actively engage in TB control.Statistical analyses; list of global positioning system (GPS)\u2013collared adult wild boars; details of GPS-collared wild boar found dead; survival curves and causes of death; and monthly death distribution of GPS-collared wild boars."} +{"text": "Reciprocal genomic disorders (RGDs) represent a unique class of recurrent genomic variation that offer insight into highly dosage sensitive regions of the morbid human genome. However, the genomic architecture mediating RGDs, namely non-allelic homologous recombination (NAHR) of flanking segmental duplications, has rendered these genomic segments recalcitrant to conventional model studies. We recently developed a novel CRISPR method that leverages the homology of segmental duplications and efficiently generates large microdeletions and microduplications that mimic NAHR in humans, including ablation or duplication of one copy equivalent of the segmental duplications. Here, we explore the functional consequences of 16p11.2 RGD in iPS derived neuronal models and across mouse tissues.We generated CRISPR-engineered 16p11.2 RGD models against an isogenic iPSC background and performed transcriptome profiling in iPSC-derived neural stem cells (NSCs) and induced neurons (iN) . We then integrated these data with RNAseq from 306 libraries from multiple tissues in 70 mouse models of reciprocal deletion and duplication of the syntenic 7qf3 region .In ongoing analyses, weighted-gene correlation network analysis (WGCNA) identified co-expression modules that were significantly enriched for 16p11.2 genes, evolutionarily constrained genes, genes robustly associated with autism spectrum disorder and developmental disorders (DDD). Pathway analyses within modules discovered enrichment of genes critical to synaptic formation and neural connectivity as well as the protocadherin gene family. Network analyses specific to brain tissues within modules further identified a convergence on highly connected, or \u2018hub\u2019 genes, on Wnt signaling, including Ctnnb1 and Ctnnd1. The module was also again enriched for ASD loci , constrained genes and brain specific genes from the Human Protein Atlas.These studies suggest the functional consequences of 16p11.2 RGD across models converge on transcriptional signatures associated with critical neurodevelopmental pathways and individual genes implicated in a spectrum of developmental and neuropsychiatric disorders."} +{"text": "The mechanism of telomerase re-activation in cancer had remained elusive until the discovery of frequent mutations in the promoter of the TERT gene that encodes the catalytic reverse transcriptase subunit of telomerase. We investigated the regulation of TERT expression in melanoma cell lines and our results show that promoter mutations render TERT expression dependent on MAPK activation due to oncogenic BRAF or NRAS mutations. Mutations in the TERT promoter create binding sites for ETS transcription factors. ETS1, expressed in melanoma cell lines, undergoes activating phosphorylation by ERK at Thr38 residue as a consequence of constitutively activated MAPK pathway. We demonstrate that ETS1 binds on the mutated TERT promoter leading to the re-expression of the gene. The inhibition of ETS1 resulted in reduced TERT expression. We provide evidence that the TERT promoter mutations provide a direct link between TERT expression and MAPK pathway activation due to BRAF or NRAS mutations via the transcription factor ETS1. Melanoma arises from the malignant transformation of melanocytes that involves numerous genetic alterations affecting multiple signaling pathways including MAPK (Mitogen Activated Protein Kinase), PI3K (Phosphoinositide 3-kinase), cAMP and cyclin D1/CDK4 , 3. A crin vitro. The transcription factors that have been shown to bind the sites include ETS1, ETS2, ELF1, ELF2, ETV6, p52 NF-\u03baB and GABPA, however, no study has so far shown link between TERT expression and MAPK activation ) and non-transfected cells were used as negative controls. The three plasmids were assayed separately with or without MAPK inhibitor (Trametinib) and additionally with or without Ets-1 overexpression. For Ets1 overexpression, the corresponding cells were cotransfected with 100 ng of PDEST26-ETS1 expression plasmid that expressed human Ets1 coding sequence (GenBank: AY893450). To determine the effect of MEK inhibitor, the cells were treated with Trametinib dissolved in DMSO at a final concentration of 1 \u03bcM, 6 hours after transfection. As a control, DMSO was used in other batch of cells that were not treated with Trametinib. Cells were harvested 30 hours post transfection using 1x passive lysis buffer (Promega) and reporter expression was analyzed using the Dual-Luciferase assay system (Promega). The relative ratio of firefly luminescence to renilla luminescence was calculated to normalize the variations across samples. Statistical differences were determined using two-sided Total RNA was extracted from melanoma cell lines and treated with DNase (ThermoFisher Scientific); cDNA was prepared using the Themoscript kit (ThermoFisher Scientific). TERT mRNA level was quantified by real-time PCR using the Power SYBR Green kit (applied biosystems) normalized to GAPDH. Sequences of the primers are in Cells were lysed in RIPA and the proteins were subjected to an SDS-PAGE and western blot analysis was carried out according to standard protocols using the following antibodies: TERT (Santa Cruz technology and Thermo Fisher Scientific), PhosphoERK (Sigma), ERK (Millipore), PhosphoETS1 (Sigma), ETS1 (Bethyl) and GABPA (Santa Cruz technology). Antibodies were visualized using the SuperSignal West Pico Chemiluminescent Substrate (ThermoFisher Scientific).Cells were cultured in 10-cm plates to approximately 80% confluence, and fixed with 1% formaldehyde. Chromatin Immunoprecipitation was carried out using the Pierce Agarose ChIP Kit (Thermo Fisher Scientific) following the manufacturer instructions. The genomic fragments were quantified by real-time PCR using the Power SYBR Green kit (applied biosystems) and primers surrounding the \u2013124 and \u2013146 TERT promoter mutations . Primers"} +{"text": "After publication of the original article it was bThe caption reads: \u201cComparative analyses of deep sequencing data on two closely related HIV RNP variants. a Pie charts summarizing how frequently was each amino acid descriptor picked as the best one for sites in RNP from HIV strains 1934 and 1968. b Sites shaped by isoelectric point with positive (blue) and negative (red) trends in the data for strain 1968 (PDB ID 2IQH [67])\u201d.This should instead read: \u201cComparative analyses of deep sequencing data on two closely related influenza RNP variants. a Pie charts summarizing how frequently was each amino acid descriptor picked as the best one for sites in RNP from influenza strains 1934 and 1968. b Sites shaped by isoelectric point with positive (blue) and negative (red) trends in the data for strain 1968 (PDB ID 2IQH [67])\u201d."} +{"text": "PARP1 inhibition caused accumulation of DSBs, which was associated with apoptosis in LIG4 deficient melanoma cells. Our hypothesis that olaparib is synthetic lethal with LIG4 deficiency in melanoma cells was supported by selective anti-tumor effects of olaparib used either alone or in combination with dacarbazine (DTIC) in LIG4 deficient, but not LIG4 proficient cells. In addition, olaparib combined with DTIC inhibited the growth of LIG4 deficient human melanoma xenografts. This work for the first time demonstrates the effectiveness of a combination of PARP1 inhibitor olaparib and alkylating agent DTIC for treating LIG4 deficient melanomas. In addition, analysis of the TCGA and transcriptome microarray databases revealed numerous individual melanoma samples potentially displaying specific defects in DSB repair pathways, which may predispose them to synthetic lethality triggered by PARP1 inhibitor combined with a cytotoxic drug.Cancer including melanoma may be \u201caddicted\u201d to double strand break (DSB) repair and targeting this process could sensitize them to the lethal effect of DNA damage. PARP1 exerts an important impact on DSB repair as it binds to both single- and double- strand breaks. PARP1 inhibitors might be highly effective drugs triggering synthetic lethality in patients whose tumors have germline or somatic defects in DNA repair genes. We hypothesized that PARP1-dependent synthetic lethality could be induced in melanoma cells displaying downregulation of DSB repair genes. We observed that PARP1 inhibitor olaparib sensitized melanomas with reduced expression of DNA ligase 4 (LIG4) to an alkylatimg agent dacarbazine (DTIC) treatment While melanomas can be successfully treated in the early stages, the appearance of metastasis in distant organs worsens prognosis and drops median survival below nine months . DespiteDSBs are highly cytotoxic DNA lesions caused by reactive oxygen species (ROS), ionizing radiation and genotoxic drugs . In prolIt was reported that cells deficient in BRCA1/BRCA2-mediated HR are sensitive to PARP1 inhibitors, such as the recently FDA approved olaparib due to induction of synthetic lethality . Since TLIG4) . Significant differences were found in the gene expression profiles between melanoma cells and melanocytes. In particular, all melanoma lines showed a decreased level of DNA ligase 4 Figure .Protein expression status of LIG4, RAD51, PARP1, Ku70 was determined by Western blot analysis in normal melanocytes and melanoma cell lines Figure . Both DMTo determine the influence of tested compounds on viable cell number, plasma membrane integrity was measured by cytometric analysis Figure . After tCell death was assessed by the appearance of sub-diploid fraction was detected only in approximately 7% of cutaneous melanomas in TCGA database Figure , inhibitIn addition, multiple melanoma samples displayed downregulation of at least one gene in HR pathway Figure with higDespite downregulation/mutations of DSB repair genes detected in numerous samples in TCGA and transcriptome microarray databases, melanomas typically do not respond well to DNA damaging agents. Perhaps the degree of downregulation of DNA repair genes is not strong enough to increase the sensitivity to chemotherapeutics in clinical settings. However, as suggested by this work, the effect may become clinically relevant in repair-deficient cells when a genotoxic drug is combined with PARP1 inhibitor, which further enhances DNA damage beyond a reparable threshold.In summary, PARP1 inhibitor seems to offer additional treatment opportunity to pre-selected melanomas displaying LIG4 (and/or XRCC4) deficiency. In addition, analyses of the already existing databases strongly suggest that numerous melanomas could be sensitive to personalized medicine-guided PARP1 inhibitor-mediated synthetic lethality due to their putative deficiencies in DNA repair pathways. This speculation is supported by phase II study showing almost doubled progression-free survival of the patients with metastatic melanoma treated with veliparib + temozolomide compared with placebo + temozolomide. Perhaps personalized medicine approach is necessary to pre-select patients with melanomas predisposed to synthetic lethality mediated by PARP1 inhibitor.www.horizondiscovery.com) and cultured in RPMI medium with 10% FBS (Lonza) and antibiotics at 37\u00b0C in a humidified atmosphere containing 5% CO2.Melanoma cell lines derived from surgical specimens of nodular and superficial spreading melanoma (DMBC11) were established in the Department of Molecular Biology of Cancer. The study was approved by the Ethical Commission of the Medical University of Lodz, and informed consent was obtained from all patients. Melanoma cells were cultured in Stem Cell Medium (SCM) as described elsewhere , 24. Nor5 viable cells per well in a 6-well plates one day before drug treatment. Cells were cultured with 5 \u03bcM olaparib (Selleckchem), 2 mM dacarbazine (DTIC) (Sigma Aldrich), olaparib + DTIC, or vehicle. After 48 hours, half the cell suspension from each well was taken to determine cell viability after propidium iodide (PI) staining and cell cycle analysis. Following this, 1 ml of fresh medium containing drugs at appropriate concentrations was added to the remaining cell culture for additional 72 hours of culturing.Melanoma cells and NHEMs were plated at a density of 1 \u00d7 10Melanoma cells were first incubated with compounds at indicated concentrations for 48 hours and then for 72 hours. Then, 1000 single viable cells were transferred to soft agar and clonogenic assay was performed as previously described .Flow cytometry and propidium iodide (PI) staining was used to assess changes in viability and cell distribution in cell cycle phases. Cells were analyzed using a FACSVerse flow cytometer . ModFit LT 3.3 software was used to calculate the percentage of cells in each cell-cycle phase and FACSuit software (Becton Dickinson) was used to calculate the percentages of dead cells in subG1.Melanoma DMBC11 cells were transfected with plasmid pCMV6-AC-GFP with cloned human LIG4 cDNA (OriGene Technologies) using lipofectamine 3000 (Invitrogen) according to the manufacturer's protocol. GFP+ cells were sorted after 48 hrs and used for the experiments.Microarray data was obtained from NCBI GEO and analyzed for phenotype classes proliferative, intermediate and invasive as described in Widmer et al . Microar\u00ae Real-Time PCR Master Mix (Life Technologies) and Agilent Technologies Stratagene Mx300SP working on MxPro software. TaqMan probes (Life Technologies) were used to analyze 8 genes whose products are essential for DSB repair pathways , and 18S RNA (Life Technologies) was included as the reference gene. The cycling parameters were 95\u00b0C for 10 minutes, 30 cycles of 95\u00b0C for 15 seconds and 60\u00b0C for 60 seconds.Isolation and purification of RNA was performed using total RNA isolation kit (A&A Biotechnology). Subsequently, RNA was transcribed into cDNA using SuperScript II Reverse Transcriptase . qRT-PCR was performed using TaqManCell lysates were obtained by incubating a cell pellet with RIPA buffer for 30 minutes. Lysates were than resolved by SDS-PAGE. The proteins were transferred onto an Immobilon-P PVDF membrane (Millipore), which were blotted overnight with primary antibodies recognizing GAPDH, DNA LIG4 (Santa Cruz Biotechnologies), Ku70, RAD51 or PARP1 (ThermoFisher Scientific). This was followed by 1 h incubation with secondary antibodies conjugated with HRP .Cell lines DMBC11, DMBC12 and NHEMs were cultured with vehicle or with drugs on black 96-well plates with a clear bottom. Analysis of the level of phosphorylated histone \u03b3-H2AX was performed using an H2AX Phosphorylation Assay Kit . Bleomycin at 35 \u03bcM for 30 min was used as a control.Cells were cultured with vehicle or drugs for 48 hours and analyzed by neutral version of comet assay to detect DSBs as described before with modifications . Briefly5 melanoma cells previously suspended in Matrigel. After 4 days tumor-bearing mice were randomly assigned into four groups; untreated, and treated intraperitoneally either with olaparib , DTIC or olaparib with DTIC (same dosing as in monotherapy) for 24 days. After the end of experiment tumors were collected and weighted. The study was approved by the local Ethical Committee.24 NSG mice were injected subcutaneously under the right scapula with 1\u00d710"} +{"text": "Neisseria gonorrhoeae isolates, however, provides us with an opportunity in which WGS can be mined for AMR determinants.Antimicrobial resistance (AMR) threatens our ability to treat the sexually transmitted bacterial infection gonorrhoea. The increasing availability of whole genome sequence (WGS) data from Neisseria database (http://pubmlst.org/neisseria). AMR genotypes were identified in WGS from 289 gonococci for which MICs against several antimicrobial compounds had been determined. Whole genome comparisons were undertaken using whole genome MLST (wgMLST).Chromosomal and plasmid genes implicated in AMR were catalogued on the PubMLST Clusters of isolates with distinct AMR genotypes were apparent following wgMLST analysis consistent with the occurrence of genome wide genetic variation. This included the presence of the gonococcal genetic island (GGI), a type 4 secretion system shown to increase recombination and for which possession was significantly associated with AMR to multiple antimicrobials.N. gonorrhoeae populations evidenced by the wgMLST clusters seen here. Genomic islands offer selective advantages to host bacteria and possession of the GGI may, not only facilitate the spread of AMR in gonococcal populations, but may also confer fitness advantages.Evolution of the gonococcal genome occurs in response to antimicrobial selective pressure resulting in the formation of distinct \u2022Gene-by-gene annotation of WGS combined with whole genome MLST (wgMLST) analyses.\u2022Provision of an online catalogue of gonococcal antimicrobial resistance (AMR) genes.\u2022Gene-by-gene analysis found an association of AMR with a T4SS known to enhance HGT.\u2022The T4SS may facilitate the spread of AMR but may also confer fitness advantages. Neisseria gonorrhoeae, the aetiological agent of the sexually transmitted disease gonorrhoea, annually causes an estimated 108 million cases globally.N. gonorrhoeae strains have developed resistance to multiple classes of antibiotics.gyrA and/or the DNA topoisomerase gene, parC.ponA and penA respectively are essential in the final stages of peptidoglycan synthesis involved in cell wall assembly. Beta-lactams such as penicillin and cephalosporin target PBP1 and PBP2 inhibiting cell wall synthesis; however, non-synonymous mutations combined with recombination alter the antibiotic target thereby limiting beta-lactam activity.mtrR efflux pump repressor gene and/or its associated promoter resulting in over-expression of the MtrCDE efflux pump,porB1b .tetM genes which facilitate penicillin and/or tetracycline resistance.12Gonococci become resistant to antibiotics through spontaneous mutation and/or horizontal genetic transfer (HGT) with resistance conferred through all known mechanisms including antimicrobial inactivation, antimicrobial target\u00a0alteration as well as increased export and decreased uptake of antimicrobial compounds.penA mosaic alleles have been identified in several WGS studies undertaken in N. gonorrhoeae.PubMLST.org/neisseria website archives and annotates, at the time of writing, >7000 WGS data from multiple Neisseria species including N. gonorrhoeae.N. gonorrhoeae dataset identifying distinct gonococcal populations clustering by AMR genotype indicative of the presence of additional genomic elements associated with AMR. This included a type 4 secretion system (T4SS) also known as the gonococcal genetic island (GGI) which is known to enhance HGT through the secretion of single stranded DNA.Advances in sequencing and bioinformatics technology provide rapid and automated analysis of whole genome sequence data (WGS) and understanding antimicrobial resistance (AMR) using WGS is likely to become essential in combatting AMR. For example, associations between resistance to the third generation cephalosporin, cefixime and possession of de novo using VELVET in combination with VELVETOPTIMISER as previously described.www.pubmlst.org/neisseria.WGS data from published isolate collections included: i) 236 isolates collected from sentinel public STD clinics by the US Centers for Disease Control and Prevention Gonococcal Isolate Surveillance Project; and ii) 53 isolates of diverse origin dating from the 1980s to 2011. Isolates had been analysed for antimicrobial minimum inhibitory concentrations (MICs) to several antibiotics.PubMLST.org/neisseria website which runs the BIGSdb genomics platform.N. gonorrhoeae isolates FA1090 (accession number NC_002946) and MS11 (accession number NC_022240) were employed.WGS data were compared using the genome comparator tool, implemented within the N. gonorrhoeae isolate MS11 (Accession number AY803022) was used as a reference. It is composed of 62 open reading frames and, sequences from each of these were defined in the database and the gene name, penA. As alterations in promoter regions located upstream of specific loci have been found to increase antibiotic resistance,pro suffix (for promoter) followed by the corresponding locus prefix for the adjacent gene to differentiate them from coding sequences, e.g. proNEIS1635 ,tetM gene. Sequences from the beta-lactamase plasmid conferring resistance to beta-lactams were retrieved from plasmid pSJ5.2 containing blaTEM1, and defined as NEIS2357\u2013NEIS2360 (DQ355980) with NEIS2357 designating the blaTEM gene software package and subsequently viewed using Tablet.The BIGSdb software includes \u2018autotagger\u2019 and \u201cautodefiner\u201d tools which scan deposited WGS against defined loci identifying alleles greater than or equal to 98% sequence identity. This process runs in the background and automatically updates isolate records with specific allele numbers, marking regions on assembled contiguous sequences (contigs) for any of the defined loci. Loci with sequence identity <98% were manually checked and curated. Using the molecular evolutionary analysis software MEGA v6, deduced amino acid sequences were aligned identifying polymorphic sites associated with antimicrobial resistance and enabling alleles containing these mutations to be detected .22 Four MIC cut-offs, guided by the US GISP antimicrobial susceptibility criteria, were defined for each antimicrobial compound . PhenotypenA (NEIS1753) allele 266 and isolates exhibiting resistance to multiple antimicrobial compounds as previously identified by Grad et\u00a0al.; this cluster also contained the GGIpenA (NEIS1753) allele 289 . A signiNeisseria meningitidis isolates from clonal complex ST-11 as well as commensal Neisseria.27Another group of isolates, previously identified as cluster 2 by Grad et\u00a0al. but indicated here as cluster 8, were ST-1580, contained NEIS1753 allele 266 as well as the GGI but were susceptible to ciprofloxacin.ponA (NEIS0414) associated with resistance to beta-lactam compounds were identified in 203/289 (70%) isolates with allele 13 the most predominant (penA (NEIS1753) alleles 266 and 281 contained penA mosaic motif XXXIV, which is associated with reduced susceptibility to third generation cephalosporins; however, penA allele 281 contained an additional non-synonymous mutation (D101\u00a0\u2192\u00a0E) found in one isolate only, GCGS126. This isolate had a cefixime MIC >0.125\u00a0\u03bcg/ml but did not possess mutations conferring resistance in any other AMR-associated loci (penA (NEIS1753) allele 266 was found in 122/289 (42%) isolates; however, 26/122 (21%) did not have mutations associated with resistance in other AMR loci. Although these isolates exhibited reduced susceptibility to cefixime, they did not have resistant phenotypic MIC values to any of the other antimicrobials . Penicilted loci . penA allele associated with decreased susceptibility to beta-lactams and tetracycline with AMR conferred through non-synonymous substitutions in loop III of PorB.pilQ (NEIS0408) associated with decreased susceptibility to cefixime and ceftriaxone,penA (NEIS1753), mtrR (NEIS1635) and porB1b (NEIS2020) mutations and therefore were susceptible to these compounds. Amino acid mutation S341\u00a0\u2192\u00a0N in pilQ , not associated with increased resistance to cephalosporins, was found in 275/289 (95%) isolates.Most isolates, 282/289 (98%) contained the blaTEM genes have been described with blaTEM1 the most commonly found followed by blaTEM135.blaTEM) alleles 3 and 9 were blaTEM1 and were found in 15/21 (71%) isolates while allele 2 designated blaTEM135 and was found in 6/21 (29%) isolates. There were two NEIS2210 (tetM) alleles with allele 1 found in 7/19 (37%) isolates and allele 2 in 12/19 (63%) isolates isolates containing the beta-lactamase plasmid and 19/289 (6%) the TetM conjugative plasmid. Divergent isolates .gyrA (NEIS1320) conferring resistance to fluoroquinolones, with allele 14 the most predominant . In parC (NEIS1525), 184/289 (64%) isolates contained amino acid substitutions at residue 87 (S87\u00a0\u2192\u00a0R) only, with allele 104 the most predominant and found in association with gyrA (NEIS1320) allele 14. All of these isolates were resistant to ciprofloxacin (parE (NEIS1600).Most isolates, 176/289 (61%), were found with mutations S91\u00a0\u2192\u00a0F and D95\u00a0\u2192\u00a0G in floxacin . None ofrpsE ribosomal protein S5 (NEIS0149) was not found.rpsE (NEIS0149) allele 83 or mutation C1186\u00a0\u2192\u00a0T in 16S rRNA allele 1538.Of the previously reported mutations associated with macrolide resistance, mutation C24\u00a0\u2192\u00a0P identified in proNEIS1635 allele 3)penA (NEIS1753), ponA (NEIS0414) and porB (NEIS2020). Four isolates were found with an A\u00a0\u2192\u00a0C substitution in the promoter region, proNEIS1635 allele 4, with these isolates also containing premature stop codons in mtrR gene consistent with putative non-functional MtrR proteins. These were associated with resistant MIC to azithromycinGAAT-3\u2032) upstream of macA (proNEIS0488) and no significant mutations were found in the transcriptional regulator, NEIS0374 (farR).norM) may occur when a T\u00a0\u2192\u00a0C nucleotide occurs in the \u221235 box in the promoter region (proNEIS0763) (TTGACG to CTGACG)The adenine deletion in the 13bp promoter region associated with increased expression of the MtrCDE efflux pump was found in 178/289 (62%) isolates into the extracellular environment .17 T4SSsydhB (NEIS2281) and ydcA (NEIS2282), located in the GGI and ydhA (NEIS2289) can be found which may enable within host competition between plasmids consistent with the low prevalence of isolates here possessing both a plasmid and the GGIExpansion of distinct gonococcal populations may also be promoted through the activity of toxin\u2013antitoxin (TA) subunits encoded by the genes, the GGI . TA are i.e. isolates had susceptible phenotypes despite the presence of resistant genotypes). These correlated with some of the lower NPV scores obtained for penicillin and tetracycline and the Oxford Martin School, University of Oxford (H2RXJo00). MCJM was supported by the Wellcome Trust (087622). YHG was supported by the National Institutes of Health (K08-AI104767-01). DLT supported by the CDC and CDC's Office of Advanced Molecular Detection (AMD-18).This study was jointly funded by a The authors declare no competing interests."} +{"text": "To evaluate the angle closure scoring system (ACSS) for stratifying primary angle course disease.g) and lens thickness/axial length ratio (LT/AL), cup disc ratio and baseline intraocular pressure (IOP) to give total score (ACSSt).This observational cross sectional institutional study included patients with primary open angle glaucoma suspects (n = 21) and primary angle closure disease . Two independent examiners blinded to clinical details, graded good quality pre-laser goniophotographs of the patients incorporating quadrants of peripheral anterior synechieae (PAS), non-visibility of posterior trabecular meshwork (PTM) and blotchy pigments (ranging from 1\u20134 quadrants), iris configuration, angle recess and PACG (Odds ratio = 1.6(95%CI-1.19\u20132.2) predicted need for single medicines while ACSSg scores >14 and 19 predicted need for \u22652 medicines in PAC and PACG eyes, respectively. The LT/Al ratio, IOP score or cup disc score did not influence the need for medical treatment independently.The ACSS can be a useful clinical adjunct to the ISGEO system to predict need for medicines and prognosticate each stage more accurately. The PAC and PACG was now stratified according to gonioscopic sum scoresACSSg<25Evaluating the difference further in ISGEO tier of PAC eyes, 8 required 1 or 2 medicines with none of the eyes requiring >2 medicines for IOP control. We found significant difference in those requiring no medicine after LPI versus those that required medicines for IOP control, Tables Evaluating differences in PACG eyes requiring different number of medications after iridotomy, the sum scores of gonioscopic parameters was significantly greater in PACG eyes requiring >2 medicines, The sum ACSSg>12 and 14 in PAC (odds ratio = 2.7(95% CI-1.7\u20135.9) and PACG(Odds ratio = 1.6(95%CI-1.19\u20132.2) predicted need for single medicines while sum ACSSg scores >14 and 19 predicted need for 2 or more medicines in PAC and PACG eyes, respectively, Evaluating now the result of adding LT/AL ratio and baseline IOP scores, there was minimal change in total ACSStscores as compared to sum scores of gonioscopic parameters alone . UnivariIn this study, definite difference in scores of angle scoreswere seen between PAC and PACG eyes as well as between eyes requiring 1 or more medicines within the same ISGEO group of PAC and PACG eyes. This reflects that the current staging system of PAC and PACG is inadequate in appropriately stratifying or prognosticating eyes since there may be more stages in each ISGEO stage of PAC and PACG during the evolution or progression of angle closure disease. The sum ACSSg was independently associated with need for >1 medicines with minimal change with addition of other parameters.Caveats of the pre-existing scoring system: Our scoring system not only identified PACG eyes requiring one or two medicines but also identified significant overlap of scores between PAC eyes at one stage and PACG eyes in the preceding stage. This reflects non-linear progression of PAC and PACG eyes at each stage and reiterates the importance of scoring systems for prognosticating PAC eyes or PACG eyes after initial definitive treatment. Currently, the ISGEO system, globally used in epidemiological surveys, is also used for clinical practise which has been known to be inappropriate on several fronts. Though sImportance of adequate baseline staging: It is accepted that anatomical factors play an important role in determining the \u201ccrowdability\u201d of the angle tough the results across ethnic populations have not been concordant.,25,26 MaThere is considerable debate of the ISGEO system failing to incorporate APAC eyes into a separate group.,36 PatieThis study was limited to a subset of patients seen at a single tertiary centre. Since we wanted to evaluate the use of this scoring system in established PACD, we included PAC and PACG eyes in this study. Dynamic changes under physiologic conditions cannot be incorporated into this scoring system which is based on clinical gonioscopic documentation during office visits. Nevertheless, this scoring system can be a useful addendum to the ISGEO system of PACD classification which may help prognosticate each stage of PAC and PACG quantitatively and accurately.S1 Fig(TIF)Click here for additional data file.S2 Fig(TIF)Click here for additional data file.S3 Fig(TIF)Click here for additional data file.S4 Fig(TIF)Click here for additional data file.S1 Table(PDF)Click here for additional data file.S2 Table(PDF)Click here for additional data file.S3 Table(PDF)Click here for additional data file."} +{"text": "Here, we present a genome-wide association study (GWAS) to identify candidate loci conferring Mn toxicity tolerance in rice (Oryza sativa L.). A diversity panel of 288 genotypes was grown in hydroponic solutions in a greenhouse under optimal and toxic Mn concentrations. We applied a Mn toxicity treatment at twelve days after transplanting. Mn toxicity caused moderate damage in rice in terms of biomass loss and symptom formation despite extremely high shoot Mn concentrations ranging from 2.4 to 17.4 mg g-1. The tropical japonica subpopulation was more sensitive to Mn toxicity than other subpopulations. Leaf damage symptoms were significantly correlated with Mn uptake into shoots. Association mapping was conducted for seven traits using 416741 single nucleotide polymorphism (SNP) markers using a mixed linear model, and detected six significant associations for the traits shoot manganese concentration and relative shoot length. Candidate regions contained genes coding for a heavy metal transporter, peroxidase precursor and Mn2+ ion binding proteins. The significant marker SNP-2.22465867 caused an amino acid change in a gene (LOC_Os02g37170) with unknown function. This study demonstrated significant natural variation in rice for Mn toxicity tolerance and the possibility of using GWAS to unravel genetic factors responsible for such complex traits.Manganese (Mn) is an essential micro-nutrient for plants, but flooded rice fields can accumulate high levels of Mn Plants were grown in hydroponic culture for 33 days. An optimum rice growing environment was maintained in the greenhouse under natural light supplemented with artificial lighting for 12 hours a day to ensure a minimum light intensity of 19.5 kilo lux, 29/22\u00b0C day/night temperature, and 49/65 percent day/night relative humidity. Twelve-day-old rice seedlings were first transplanted into 60 liter tanks filled with half strength modified Yoshida solution in TASSEL 5.0 analysis. The latter results are presented as supplementary data annotated for abiotic stress response were present in 66 kb window with significant SNP-2.17092835 (Table B in 10 (p) value were detected on chromosome 7 because the significant SNPs were in strong LD . NotablyIn this study we employed a HDRA composed of 700 k SNPs for a suDro1 [Pup1 [Among the candidate genes identified in this study, a SNP variant detected for RSL causing an amino acid substitution was located right in the coding sequence of LOC_Os02g37170 . AlthougDro1 or Pup1 o1 [Pup1 , were unOsNRAMP5 (LOC_Os07g15370) is known to be involved in root to shoot translocation of Mn in rice [OsNRAMP1 (LOC_Os07g15460), an orthologue of AtNRAMP1 which has been characterized as a high affinity Mn transporter regulating Mn uptake in Arabidopsis [Some of the candidate genes contained in candidate loci can be assigned to typical adaptive strategies employed by plants. The transport of Mn between plant organs plays an important role in Mn tolerance. in rice ,56 and wbidopsis .indica) accumulated more Mn in rice shoots than the sensitive one [2+. These genes are highly expressed in both roots and shoots especially in early vegetative stages (http://ricexpro.dna.affrc.go.jp). Moreover, proteins with metal binding domains are known to confer heavy metal toxicity tolerance [2+ binding proteins were present in the candidate loci for SMC in this study. Likewise, antioxidant activity is important for the scavenging ROS, which can arise from reactions involving transition metals such as Mn [OsGRX genes were constitutively expressed in shoots in vegetative growth stages (http://ricexpro.dna.affrc.go.jp). GRX is involved in maintaining glutathione dependent peroxidase activity as part of the antioxidant system during oxidative stress in plants [Shoot tolerance is generally referred to as the ability to tolerate high concentrations of elements in aboveground tissue through inclusion mechanisms. Mn shoot tolerance was illustrated in a bi-parental QTL mapping study, where the tolerant parent Click here for additional data file.S2 FileTable A\u2013Table C.(XLSX)Click here for additional data file."} +{"text": "Despite the pre-clinical and clinical success of KP1019, the mode of action and various factors capable of modulating its effects are largely unknown. Here, we used transcriptomics and genetic screening approaches in budding yeast model and deciphered various genetic targets and plethora of cellular pathways including cellular signaling, metal homeostasis, vacuolar transport, and lipid homeostasis that are primarily targeted by KP1019. We also demonstrated that KP1019 modulates the effects of TOR (target of rapamycin) signaling pathway and induces accumulation of neutral lipids (lipid droplets) in both yeast and HeLa cells. Interestingly, KP1019-mediated effects were found augmented with metal ions (Al3+/Ca2+/Cd2+/Cu2+/Mn2+/Na+/Zn2+), and neutralized by Fe2+, antioxidants, osmotic stabilizer, and ethanolamine. Additionally, our comprehensive screening of yeast histone H3/H4 mutant library revealed several histone residues that could significantly modulate the KP1019-induced toxicity. Altogether, our findings in both the yeast and HeLa cells provide molecular insights into mechanisms of action of KP1019 and various factors that can alter the therapeutic efficiency of this clinically important anticancer drug.KP1019 ([trans-RuCl The failure to decipher and target the multiple underlying pathways of molecular carcinogenesis is a major challenge for anticancer drug development. Hence, recent efforts are being made towards developing additional therapeutic molecules targeting the plethora of cancer mechanisms to achieve higher therapeutic efficiency (specific to cancer cells) with lesser or no adverse effects ) has demonstrated most promising anticancer activity with minimal side-effects in the undergoing clinical trials ) was developed to improve its solubility. It has shown promising results in phase-I trials against a variety of solid tumors cell viability assay as described earlier *100. \u2018ODt\u2019 represents the mean absorbance of treated cells at 570nm, \u2018ODc\u2019 represents the mean absorbance of solvent (DMSO) treated control cells at 570nm, and \u2018ODb\u2019 represents the mean absorbance of respective well at 690nm. Additionally, the abovementioned treatments were repeated in a 24-well plate . After 36h, the morphology of HeLa cells was recorded using an inverted microscope .The viability of cells was calculated as % of control as following: % of control= [(ODSaccharomyces cerevisiae strains used in this study were listed in pPW344 (URA3), pHL126 (URA3) plasmids using the standard lithium acetate procedure [The rocedure and the Growth sensitivity assay was performed to examine the effect of KP1019 on the growth of yeast cells as described previously . In brie600) values measured at a regular interval of 30min for indicated period using a plate reader (Eon\u2122 Microplate Spectrophotometer) [For growth curve analysis, exponentially growing yeast cells were treated in duplicate with either DMSO (control) or indicated doses of KP1019, ETA, and GSH alone or in combination and then seeded in a 96-well cell culture plate (SPL Life Sciences Ltd.). Growth curves were constructed for each treatment using representative optical density (ODtometer) .in vitro transcription reaction and then hybridized to Yeast Genome 2.0 GeneChip Arrays (GPL2529) by following a standard Affymetrix protocol. Afterward, GeneChips were washed, stained in the Fluidics Station 450 (Affymetrix) and scanned using the GeneArray 30007G microarray scanner. The data sets were extracted from all CEL (raw intensity) files and submitted to NCBI's Gene Expression Omnibus (GEO) repository with a GEO Series accession number of GSE76985 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE76985).The exponentially growing wild-type yeast (1588-4C) cells were treated with KP1019 (50\u03bcg/ml) or equivalent DMSO (solvent control) for 3h and then harvested. Total RNAs were isolated by heat/freeze Phenol method as described earlier and asseThe raw signals (CEL) in each array were processed for background adjustment, normalization followed by log transformation and summarization of probe sets using RMA (Robust Multi-array Average) algorithm in GeneSpring GX 12.6 expression analysis software . Differentially expressed genes (DEG\u2019s) whose expression altered (induced or repressed) significantly by 1.5-fold (>1.5) in KP1019 treatment compared to DMSO (control) were determined by moderated t-test (p<0.05). Functional classification and Gene Ontology (GO) enrichment analysis of DEG's were performed using standard tools described in the HAC1-S and ACT1 . The PCR amplicon products were electrophoresed, stained with ethidium bromide, and photographed [Total cellular RNAs were extracted from yeast cells by heat/freeze phenol method as described earlier . 1\u03bcg of ographed .Whole-cell protein extracts were obtained by 20% Trichloroacetic acid (TCA) precipitation method as described previously . The pro\u03b2-galactosidase assay was performed to monitor the expression levels of UPRE-lacZ reporter gene as described previously [600: 0.8-1) carrying 2\u03bc UPRE-lacZ reporter plasmid [600 was determined. The \u03b2-galactosidase activity was measured by permeabilizing the cells in 1ml of Z buffer and using ONPG as a substrate. The galactosidase activity was expressed in terms of Miller units as described earlier [The eviously . Briefly Jensen) were tre earlier .ss-dsRed-HDEL reporter [pHL126 plasmid [pHL126 plasmid were grown till exponential phase (OD600: 1-1.2) and left untreated (DMSO control) or treated with KP1019 . Then the localization of Sfp1 was analyzed after 1h and 3h of KP1019 treatment using 63x oil-immersion objective lens of ZEISS ApoTome.2 microscope provided with the appropriate filter.To investigate the effects of KP1019 on neutral lipids (LDs), ER architecture, and Sfp1 localization in yeast, we employed BODIPY 493/503 dye , cells hlear ER) , and pHLd Shore) , respectTo test the effect of KP1019 on LDs in mammalian cells, HeLa cells were seeded in two-chambered slides using DMEM medium and left undisturbed for 24h. The cells were left untreated or treated with KP1019 for 36h. The cells were then stained with 5\u03bcM of BODIPY 493/503 dye after 30min fixation with 3.7% formaldehyde as indicated earlier . The celTo assess the uptake of KP1019 by yeast cells, intracellular Ru levels were determined using ICP-MS as described previously . A detai600 of 0.1, and then replica spotted onto SC-agar tray plates that were supplemented without (DMSO control) or with 50\u03bcg/ml of KP1019 (for sensitivity), 100\u03bcg/ml of KP1019 (for resistance). The plates were incubated at 30\u00b0C and imaged after 48h. The growth fitness of mutants was compared to their respective wild-type cells spotted on the same plate and also to their corresponding fitness on the control (DMSO) plate. The mutants with decreased and increased fitness (in the presence of KP1019) compared to wild-type cells were assigned as KP1019 \u2018sensitive\u2019 and \u2018resistant\u2019 respectively. For further confirmation, KP1019 sensitive and resistant mutants were validated by both the growth assays and growth curves as described above.All the synthetic yeast histone H3/H4 library mutants were pre-cultured in SC liquid medium (Uracil dropped) till the saturation in a 96-well plate. The cell density of cultures was normalized to an ODEach of KP1019 sensitive and resistant mutants was scored using the similar methodology described by Rizzardi et al. . BrieflyFor assessing the functional role of histone H3/H4 residues whose mutants exhibited altered KP1019 tolerance, we extracted information about their position in the nucleosome, status of PTM, and associated phenotypes from the HistoneHits database . The hisThe target genes of KP1019 that were obtained here through yeast growth assays were used to identify the corresponding human homologs and their role in different human diseases using YeastMine tool . Diseaset-test . P<0.05 considered as significant compared to control.All the quantitative results are shown as Mean\u00b1SEM. The details about the number of independent experiments and experimental repeats are provided in the corresponding figure legends. Unless otherwise stated, statistical significance was assessed by performing Student's"} +{"text": "Because pediatric antineutrophil cytoplasmic antibody-associated vasculitis is rare, management generally relies on adult data. We assessed treatment practices, uptake of existing clinical assessment tools, and interest in pediatric treatment protocols among rheumatologists caring for children with granulomatosis with polyangiitis (GPA) and microscopic polyangiitis (MPA).A needs-assessment survey developed by an international working group of pediatric rheumatologists and two nephrologists was circulated internationally. Data were summarized with descriptive statistics. Pearson\u2019s chi-square tests were used in inferential univariate analyses.p\u00a0=\u00a00.0190, n\u00a0=\u00a0139), those with >5\u00a0years of independent practice experience , and those who had seen >10 children with GPA/MPA in their careers . Respondents who had treated >10 patients were also more likely to continue maintenance therapy for at least 24\u00a0months . Ninety six percent of respondents believed in a need for pediatric-specific treatment guidelines; 46% supported adaptation of adult guidelines while 69% favoured guidelines providing a limited range of treatment options to allow comparison of effectiveness through a registry.The 209 respondents from 36 countries had collectively seen ~1600 children with GPA/MPA; 144 had seen more than two in the preceding 5 years. Standardized and validated clinical assessment tools to score disease severity, activity, and damage were used by 59, 63, and 36%, respectively; barriers to use included lack of knowledge and limited perceived utility. Therapy varied significantly: use of rituximab rather than cyclophosphamide was more common among respondents from the USA contains supplementary material, which is available to authorized users. Antineutrophil cytoplasmic antibody- (ANCA-) associated vasculitis (AAV) describes the subset of vasculitides primarily involving small vessels: granulomatosis with polyangiitis , microscopic polyangiitis (MPA), eosinophilic granulomatosis with polyangiitis , and renal-limited pauci-immune glomerulonephritis . Althoughttps://clinicaltrials.gov/ct2/show/NCT02006134) has allowed collection of clinical data and biological samples to ARChiVe. While pediatric clinical trials remain difficult, an international registry such as ARChiVe might provide the opportunity to compare effectiveness of a limited range of standardized treatment options and ultimately generate evidence-based guidelines. This strategy has been pursued for uvenile dermatomyositis, another rare pediatric rheumatic disease [A 2005 survey of Childhood Arthritis and Rheumatology Research Alliance (CARRA) members elicited overwhelming consensus on the need to study childhood-onset vasculitis independently from adult disease . ARChiVeAs a first step toward developing treatment guidelines for pediatric AAV, we sought to better understand the diversity of beliefs and practices associated with current care. We conducted an international needs assessment survey of physicians involved in the diagnosis and management of children with GPA and MPA, with the following aims: (1) to assess the level of community experience with pediatric AAV; (2) to assess uptake of existing classification criteria, clinical scoring tools, and treatment guidelines; (3) to assess the extent of variation in current treatment practices; and (4) to determine interest in and capacity for use of pediatric treatment protocols.A survey draft developed by the Pediatric Rheumatology group and a nephrologist at British Columbia Children\u2019s Hospital was finalized with input from the Vasculitis Working Groups of CARRA and PRES. The survey comprised 47 predominantly categorical multi-choice questions in three sections , the Australian Pediatric Rheumatology Group, the PRES-CARRA Vasculitis Working Group, and the Pediatric Rheumatology Bulletin Board (ped-rhe-list@mcmaster.ca) were invited by email to complete the web-based survey . The survey was administered using REDCap electronic data capture tools hosted ap-values are reported. Analysis was performed using Microsoft Excel Version 14.6.7 and Prism Version 5.0a .Descriptive statistics were used to quantify response frequencies and means. Unless otherwise specified, frequencies are reported relative to the total number of respondents completing each question. Odds ratios (OR) were determined as measures of association. Limits of 95% confidence intervals of OR are reported in square brackets. Inferential univariate analysis with two-sided Pearson\u2019s chi-square tests and Benjamini-Hochberg correction for multiple comparisons was used to test 11 hypotheses with a false discovery rate of 5% . CorrectOf 216 respondents opening the survey, 209 completed it, yielding an estimated response rate of approximately 40%. Sixty-five (31%) chose to exit as they had not cared for two or more patients with GPA/MPA in the preceding 5 years. The 144 respondents completing the full survey practiced in 36 countries, predominantly the USA (48%), with others from Canada (13%), Italy (4%), Australia (3%), Germany (4%), Turkey (4%), United Kingdom (4%), Brazil (2%), and Sweden (2%). Most respondents (92%) belonged to one or more national or international rheumatology organizations, including CARRA (56%), PRES (32%), CAPRI (10%), the PRES-CARRA Vasculitis Working Group (10%), and others (18%). Approximately half of respondents had practiced for less than 10\u00a0years after formal training (21% for less than 5 years and 26% for 5-10\u00a0years); among the rest, 20% each had practiced for 10-20 and 20-30\u00a0years and 10% for 30-40\u00a0years. The combined lifetime experience of respondents was at least 1600 patients with GPA/MPA (some shared): 47% had seen fewer than ten patients, 39% had seen 10-20, and only 14% had seen more than 20. The majority of respondents (61%) had seen five or fewer patients with a new diagnosis of GPA or MPA within the past 5 years .Most respondents belonged to group practices, sharing diagnostic and treatment decisions for all patients (46%) or managing patients independently while sharing on-call responsibilities (41%). A minority worked in a solo practice (7%) or in another practice arrangement (6%), predominantly hospital-based. For patients who had also been assessed by a nephrologist, the majority of respondents reported making collaborative treatment decisions, 30% in a combined nephrology/rheumatology clinic and 37% in separate clinic settings. Some reported variation in the responsible subspecialty depending on factors such as who had seen the patient first (18%), while 13 and 2% of respondents reported independent management by rheumatology or nephrology, respectively.There were between 129 and 144 responses 89-100%) to subsequent individual questions regarding diagnosis and management. While the majority (73%) reported that they always sub-classify AAV as either GPA or MPA, 26% endorsed only sometimes sub-classifying. Table 9-100% toThe proportions of respondents using formal tools to assess disease severity, activity, and damage are shown in Table p\u00a0=\u00a00.7206, n\u00a0=\u00a0131), although respondents practicing outside the USA and Canada were more likely to want to participate than those within . Most respondents supported consensus guidelines drafted by an expert group (69%) and believed in the need for a limited range of options to allow for comparative outcome assessment through a clinical registry (63%). Slightly less than half (46%) felt modification of recommendations for adult disease was an acceptable method for generating pediatric treatment guidelines.A majority of respondents (53%) reported using a combination of resources to guide treatment decisions, most commonly EULAR/EUVAS recommendations (24%). A minority used site-specific standardized protocols (7%); others used pediatric textbook recommendations (4%), individualized approaches according to personal interpretation (4%), or advice from colleagues (6%). The majority of respondents (96%) believed in a need for pediatric treatment guidelines for GPA/MPA. Over half (58%) were interested in being involved in the process of consensus guideline development \u2013 43% through an iterative survey \u2013 while others felt they did not have the time (13%) or relevant expertise (19%), or were unsure (9%). There was no association between group membership and the desire to participate in guideline development , those with greater than 5\u00a0years of experience , and those who had seen more than 10 patients with GPA/MPA in their careers .All respondents followed a remission-induction and remission-maintenance model, switching from induction to maintenance therapy within 3-6\u00a0months. In adult studies, choice of induction agent may be adjusted based on measures of disease severity, primarily to limit use of aggressive life-saving treatment \u2013 specifically cyclophosphamide \u2013 that also has significant toxicity and may be unwarranted in milder disease. Two-thirds (67%) of respondents endorsed always using more aggressive treatment for patients with severe disease, while 32% reported only sometimes choosing induction therapy based on disease severity. Table Figure p\u00a0=\u00a00.2009, n\u00a0=\u00a0139), years of practice , or total number of patients seen with GPA/MPA . The provisional choice for maintenance therapy duration of 24 (46%), 18 (19%), 36 (14%), or 12 (13%) months was associated with total patient experience: 75% of respondents who had seen more than 10 GPA/MPA patients in their careers planned for at least 24\u00a0months of therapy, compared to 52% of those who had seen fewer than 10 patients . Duration of maintenance therapy was not associated with practice location or years of practice . Concurrent corticosteroid duration initiated at induction was typically six (39%) or 12 (40%) months, with some respondents continuing for 18 (6%), 24 (4%), or 36 (0.7%) months and 11% chose variable periods according to disease severity. 93% of respondents routinely recommended plasma exchange in certain situations: severe pulmonary hemorrhage and/or rapidly progressive renal disease (76%), rapidly progressive renal disease only (8%), pulmonary hemorrhage only (5%), and with co-existent conditions (10%).The most commonly used remission-maintenance treatment was AZA, followed by MTX, MMF, and RTX Table . There wAll respondents felt that an international collaborative registry was important for comparative outcome assessment of treatment strategies for GPA/MPA, with 64% of respondents selecting \u201cvery important\u201d . Primary motivations for participation in clinical studies or collaborative registries included the potential to improve outcomes for children with AAV (91%), access to available tools and resources (58%), endorsement by a formal network of investigators (49%), potential publication authorship (46%), and association with specific research objectives (40%). Only 15% of respondents felt that a monetary stipend was a major motivation. Registry-associated resources believed to be of most value included an automated PVAS calculator (82%), an online algorithm to stage disease severity with links to corresponding treatment guidelines (68%), an automated PVDI calculator (66%), a classification tool based on entered patient data (54%), and a printable table to track patient data over multiple visits (54%). Most clinicians had resources available to assist with the use of registry data entry and clinical tools, including computer and internet access in the clinic (80% each) and a trainee or fellow (58%). Fewer had a research assistant (47%), support for review board applications (37%), technology support (33%), or a research nurse (23%). Major barriers to registry participation included insufficient support for data entry (59%), lack of time (43%), burden of ethics review board approval (32%), and lack of patients (24%).This survey emphasizes the limited individual experience among clinicians caring for children with GPA and MPA and reiterates clinicians\u2019 aspirations to study childhood-onset vasculitis independently from adult disease . InternaMost respondents believed that the distinction between GPA and MPA in patients with AAV was important for prognostication, consistent with meta-analyses of adult studies showing lower 5-year survival in MPA versus GPA and of pMultiple national and international rheumatology organizations have published treatment guidelines for management of AAV in adults \u201331. ManyConsistent with guidelines for treatment of adults with AAV, all survey respondents employed a remission-induction and remission-maintenance model. Although 25% of respondents used EULAR/EUVAS recommendations to guide their treatment decisions, many employed a combination of resources. There was significant variation in medication and dosing regimens for childhood GPA/MPA, with variable use of evidence-based recommendations from existing adult guidelines.An overwhelming majority of respondents believed in the need for pediatric-specific treatment guidelines and supported guideline development by expert group consensus. It is not simple to reconcile these aspirations with the limited uptake of recent pediatric-specific research and consensus reports, although the described lack of knowledge or familiarity \u2013 as well as unclear added benefit beyond current practice \u2013 likely contribute. In view of the high frequency of renal disease (75%) among pediatric AAV patients and the A recent European initiative to develop standards of care for pediatric rheumatic diseases, SHARE (Single Hub and Access point for Paediatric Rheumatology in Europe) , will soLimitations of this study include a sample population that likely underestimates true variability in practice given the selection bias associated with elective survey completion and respTaken together, these data suggest a need for and interest in consensus treatment guidelines for pediatric GPA and MPA. They also point to potential challenges associated with guideline development and implementation. Understanding the barriers to uptake of existing classification criteria and formal assessment tools will inform efforts to improve standardization of classification and assessment to guide therapy. Physicians\u2019 aspirations for pediatric-specific, evidence-based treatments may motivate uptake of guidelines that evolve from this needs assessment survey and, imminently, from SHARE. Consensus-derived guidelines that include a range of specific treatment options \u2013 if provided together with mechanisms for comparative effectiveness research through an international registry \u2013 may facilitate clinician engagement and ultimately lead to improved outcomes for children with AAV."} +{"text": "Escherichia coli (EPEC) pathotype represents a minor proportion of E.\u00a0coli O103 strains shed in the feces of feedlot cattle. The draft genome sequences of 13 strains of EPEC O103 are reported here. The availability of the genome sequences will help in the assessment of genetic diversity and virulence potential of bovine EPEC O103.Enteropathogenic Escherichia coli (EPEC), including the EPEC O103 serogroup (E.\u00a0coli (EHEC), EPEC strains do not carry Shiga toxin genes (stx1 and/or stx2), yet they share many important virulence genes with EHEC (eae), which together with other LEE-carried genes can cause microvillus destruction during human infection, resulting in the characteristic attaching and effacing lesions isolated from the feces of feedlot cattle in the United States (E.\u00a0coli pathotype-serogroup combinations, including EHEC O103. They will also serve as a valuable resource toward the study of EPEC O103 genome evolution.The publication of the sequences of these 13 strains will contribute to the currently limited amount of publicly available sequence data on EPEC O103. These genomes will allow for investigations into the genetic similarities and differences of EPEC O103 strains to other major This whole-genome shotgun project has been deposited at DDBJ/ENA/GenBank under the accession numbers listed in"} +{"text": "Human cytomegalovirus (HCMV) is one of the opportunistic infections associated with significant morbidity and mortality among HIV/AIDS patients especially before introduction of antiretroviral therapy (ART). Little is known regarding the humoral immune response against HCMV in relation to CD4 counts among HIV infected individuals. A total of 90 achieved sera from HIV infected patients attending Bugando Medical centre care and treatment centre (CTC) aged 18 years and above were retrieved and analyzed. Sociodemographic data were collected using structured data collection tool. Detection of specific HCMV antibodies was done using Indirect Enzyme Linked Immunosorbent Assay (ELISA). Data were analyzed by using STATA version 11. A total of 90 HIV infected patients were enrolled in the study whereby 36(40%) had immunological treatment failure. The mean age of the study participants was 39\u00b112.3 years. The Prevalence of specific HCMV IgG antibodies was 84 while the prevalence of specific HCMV IgM antibodies was 2(2.3% 95% CI: 0.8-5.4). The median CD4 counts at 6 months and 12 months on HAART were significantly high in treatment success group. At 12 months of HAART as CD4 counts increases the HCMV IgG index value was also found to increase significantly, p=0.04. Significant proportion of HIV infected individuals was infected with HCMV. Higher median HCMV IgG titers were observed among patients with immunological treatment success. There is a need to investigate humoral immune responses in HIV infected individuals in relation to CD4 counts against various infectious diseases in developing countries where most of these infections are endemic. Human cytomegalovirus (HCMV) has been known to establish latency following primary subclinical infections in immunocompetent individuals , 2. In tA total of 90 achieved sera from HIV infected patients attending Bugando Medical centre CTC aged above 18 years were retrieved and analyzed. The sera were obtained at 12 months of HAART. Sociodemographic and other relevant information were obtained from patients' records. Detection of specific HCMV IgG antibodies was done using commercial indirect enzyme-linked immunosorbent assay according to manufacturer's instructions -13. DataA total of 84 sera had specific HCMV IgG antibodies while only 2(2.3% 95% CI: 0.8-5.4) had specific HCMV IgM antibodies. Higher rates of HCMV IgG seroprevalence have been similarly observed elsewhere among HIV infected individuals , 15. In In conclusion significant proportion of HIV infected individuals were HCMV IgG sero-positive and the HCMV IgG titers significantly increased as CD4 counts increases. Further studies to investigate the humoral immune response in HIV infected individuals in relation to CD4 counts against various infectious diseases are warranted in developing countries.The authors declares that they have no competing interests."} +{"text": "TSC1 or TSC2, resulting in constitutive activation of the mechanistic target of rapamycin complex 1 pathway. The activating BRAF V600E mutation is a common genetic alteration in low grade gliomas and glioneuronal tumors, and has been reported in SEGAs as well. In the present study, we assessed the prevalence of the BRAF V600E mutation in a large cohort of TSC related SEGAs and found no evidence of either BRAF V600E or other mutations in BRAF. To confirm that these SEGAs fit the classic model of two hit TSC1 or TSC2 inactivation, we also performed massively parallel sequencing of these loci. Nineteen (19) of 34 (56%) samples had mutations in TSC2, 10 (29%) had mutations in TSC1, while 5 (15%) had no mutation identified in TSC1/TSC2. The majority of these samples had loss of heterozygosity in the same gene in which the mutation was identified. These results significantly extend previous studies, and in agreement with the Knudson two hit mechanism indicate that biallelic alterations in TSC2 and less commonly, TSC1 are consistently seen in SEGAs.Subependymal giant cell astrocytomas (SEGAs) are rare, low-grade glioneuronal brain tumors that occur almost exclusively in patients with tuberous sclerosis complex (TSC). Though histologically benign, SEGAs can lead to serious neurological complications, including hydrocephalus, intractable seizures and death. Previous studies in a limited number of SEGAs have provided evidence for a biallelic two-hit inactivation of either TSC1 encoding hamartin, or TSC2 encoding tuberin. Together these two proteins form the TSC protein complex that regulates mechanistic target of rapamycin complex 1 (mTORC1) , 4140, 4Tissue was fixed in 10% buffered formalin and embedded in paraffin. Paraffin-embedded tissue was sectioned at 6 \u03bcm, mounted on organosilane-coated slides and stained with hematoxylin-eosin (HE) for the morphological evaluation. Histological diagnosis was performed according to the 2016 WHO classification of the central nervous system . SectionBRAF including codon 600 was performed as previously described using primers TCATAATGCTTGCTCTGATAGGA and GGCCAAAAATTTAATCAGTGGA [DNA was extracted from both FFPE (n=44) and frozen (n=14) SEGA tumor samples. Since SEGA often display intratumoral hemorrhages, areas of representative tumor (identified on hematoxylin & eosin stained sections) were selected for cases in which hemorrhages, were observed within the FFPE SEGA tissue samples (n=44). Tumor DNA was extracted from 10-\u03bcm-thick paraffin sections using BiOstic FFPE Tissue DNA Isolation kit (MO BIO) according to the manufacturer's instructions. From frozen tissue samples (N=14) DNA was recovered from the organic phase following QIAzol (Qiagen) extraction of RNA and was further purified using QIAamp DNA mini Kit (Qiagen). PCR amplification for the entire extent of exon 15 of TCAGTGGA . PurifieKIAA1549-BRAF fusions in a diagnostic setting. Total RNA was extracted from frozen tissue samples using miRNeasy mini kit (Qiagen) according to the manufacturer's instructions. One microgram of total RNA was reverse-transcribed into cDNA, followed by PCR using primer sets corresponding to different KIAA1549-BRAF fusion genes and the PBGD and B2M reference genes , targeted MPS was performed using a HaloPlex custom capture array as described previously . In the http://exac.broadinstitute.org), 1000G and GnomAD [BRAF were analysed using cBio and were further assessed for functionality using 3 different in silico prediction tools: PROVEAN (http://provean.jcvi.org), SIFT (http://sift.jcvi.org) and MutationAccessor (http://mutationassessor.org) [The sequencing output was de-convoluted into individual sample reads and sorted using Picard tools . Reads wd GnomAD , 56. Varsor.org) \u201361.A second approach was used in parallel to analyze the sequence data, with capture of read calls at all positions using SAMtools Pileup, followed by custom processing in Python and Matlab to determine base call frequency at each position in each read orientation. These data were then filtered to eliminate variant calls observed in only a single read orientation, or seen in multiple samples to exclude artifacts derived from the sequencing process. All variants observed at a frequency of >1% were directly reviewed using the Integrative Genomics Viewer, to identify bona fide variant calls and exclude sequencing or alignment artifacts , 23, 26.TSC1 and TSC2 was required for the samples reported here. The median read depth for coding exons of TSC1 and TSC2 was a median of 107 (range 20 \u2013 1120) among the 31 samples.A minimal median read depth of 20x coverage for the coding exons of TSC1 and TSC2 genes that had a population allele frequency of > 0.05% in the GnomAD database. If either the mutant allele frequency for the mutation was > 55%, or the median SNP minor allele frequency for TSC1/TSC2 was < 40%, this was considered evidence of CN-LOH. LOH was assessed only in the tumor samples; normal brain tissue adjacent to the tumor, was not available.LOH was assessed using two allele frequencies: 1) at the site of mutation, using Unix grep to precisely quantify mutant vs. wild-type reads for indel mutations; and 2) at all SNPs identified in the"} +{"text": "Densitometry on paired inspiratory and expiratory multidetector computed tomography (MDCT) for the quantification of air trapping is an important approach to assess functional changes in airways diseases such as cystic fibrosis (CF). For a regional analysis of functional deficits, an accurate lobe segmentation algorithm applicable to inspiratory and expiratory scans is beneficial.We developed a fully automated lobe segmentation algorithm, and subsequently validated automatically generated lobe masks (ALM) against manually corrected lobe masks (MLM). Paired inspiratory and expiratory CTs from 16 children with CF (mean age 11.1\u00b12.4) acquired at 4 time-points with 2 kernels were segmented, resulting in 256 ALM. After manual correction spatial overlap (Dice index) and mean differences in lung volume and air trapping were calculated for ALM vs. MLM.3 for the right upper lobe to 17.41\u00b114.92cm3 for the right lower lobe. Higher differences were noted on expiratory CT. The mean differences for air trapping were even lower, ranging from 0\u00b10.01 for the right upper lobe to 0.03\u00b10.03 for the left lower lobe.The mean overlap calculated with Dice index between ALM and MLM was 0.98\u00b10.02 on inspiratory, and 0.86\u00b10.07 on expiratory CT. If 6 lobes were segmented (lingula treated as separate lobe), the mean overlap was 0.97\u00b10.02 on inspiratory, and 0.83\u00b10.08 on expiratory CT. The mean differences in lobar volumes calculated in accordance with the approach of Bland and Altman were generally low, ranging on inspiratory CT from 5.7\u00b152.23cmAutomatic lobe segmentation delivers excellent results for inspiratory and good results for expiratory CT. It may become an important component for lobe-based quantification of functional deficits in cystic fibrosis lung disease, reducing necessity for user-interaction in CT post-processing. Quantitative computed tomography (QCT) of cystic fibrosis (CF) lung disease is becoming an increasingly recognized and viable approach for evaluation of CF airway disease. Chronic progressive lung disease continues to determine more than 90% of morbidity and mortality in patients with CF \u20133. RecenThe prospective multi-center study was carried out in 36 subjects enrolled in the Novartis CF Natural History Study from 20016 children with confirmed CF at baseline were included in the study who were subjects in a joint Novartis Pharmaceutical\u2014Cystic Fibrosis Foundation Therapeutics Development Network Consortium were included for this analysis. These 16 children were selected out of 36 subjects because they were examined exactly with the same scanner in the same institution. Moreover, they differed neither in the severity of the disease nor in any other aspect from the remaining 20 participants in the joint Novartis Pharmaceutical\u2014Cystic Fibrosis Foundation Therapeutics Development Network Consortium study. Demographic data is reported in All patients underwent multidetector CT at baseline, and consecutively after 3, 12 and 24 months, comprising a total of 64 exams, i.e. 128 volumetric datasets. Exclusively non-enhanced spirometer-controlled paired inspiratory and expiratory CT was routinely performed in supine position as reported previously , 29, 30.The in-house program YACTA (version 2.7.1.3) segmented the lungs and individual lobes fully automatically on inspiratory images as employed in previous studies , 31, 32 After fully automatic lobe segmentation on inspiratory and expiratory CT datasets, the lobe masks were reviewed by a radiologist, and the masks were corrected manually for each inspiratory and expiratory acquisition to perfectly separate pulmonary lobes . This prD and J) were calculated for aforesaid groups .(PDF)Click here for additional data file.S5 Table3] calculated for fully automatic and manually corrected segmentation on inspiration B30f scans at baseline, 3, 12 and 24 months. The last column summarizes all time points. All values are separately calculated for the right upper (RUL), middle (RML) and lower lobe (RLL), the left upper lobe (LUL), the lingula (LLi), the left lower lobe (LLL), and also combining left upper lobe and lingula into one lobe (LUL+LLi). Both methods are compared in accordance with the approach of Bland-Altman giving mean differences (\u0394), limits of agreement (LoA) and two regression coefficients (Intercept / Slope and Pearson\u2019s correlation coefficient).Lobar volumes [cm(PDF)Click here for additional data file.S6 Table3] calculated for fully automatic and manually corrected segmentation on inspiration B60f scans at baseline, 3, 12 and 24 months. The last column summarizes all time points. All values are separately calculated for the right upper (RUL), middle (RML) and lower lobe (RLL), the left upper lobe (LUL), the lingula (LLi), the left lower lobe (LLL), and also combining left upper lobe and lingula into one lobe (LUL+LLi). Both methods are compared in accordance with the approach of Bland-Altman giving mean differences (\u0394), limits of agreement (LoA) and two regression coefficients (Intercept / Slope and Pearson\u2019s correlation coefficient).Lobar volumes [cm(PDF)Click here for additional data file.S7 Table3] calculated for fully automatic and manually corrected segmentation on expiration B30f scans at baseline, 3, 12 and 24 months. The last column summarizes all time points. All values are separately calculated for the right upper (RUL), middle (RML) and lower lobe (RLL), the left upper lobe (LUL), the lingula (LLi), the left lower lobe (LLL), and also combining left upper lobe and lingula into one lobe (LUL+LLi). Both methods are compared in accordance with the approach of Bland-Altman giving mean differences (\u0394), limits of agreement (LoA) and two regression coefficients (Intercept / Slope and Pearson\u2019s correlation coefficient).Lobar volumes [cm(PDF)Click here for additional data file.S8 Table3] calculated for fully automatic and manually corrected segmentation on expiration B60f scans at baseline, 3, 12 and 24 months. The last column summarizes all time points. All values are separately calculated for the right upper (RUL), middle (RML) and lower lobe (RLL), the left upper lobe (LUL), the lingula (LLi), the left lower lobe (LLL), and also combining left upper lobe and lingula into one lobe (LUL+LLi). Both methods are compared in accordance with the approach of Bland-Altman giving mean differences (\u0394), limits of agreement (LoA) and two regression coefficients (Intercept / Slope and Pearson\u2019s correlation coefficient).Lobar volumes [cm(PDF)Click here for additional data file.S9 TableAir trapping (E/I MLA) calculated for fully automatic and manually corrected segmentation on B30f scans at baseline, 3, 12 and 24 months. The last column summarizes all time points. All values are separately calculated for the right upper (RUL), middle (RML) and lower lobe (RLL), the left upper lobe (LUL), the lingula (LLi), the left lower lobe (LLL), and also combining left upper lobe and lingula into one lobe (LUL+LLi). Both methods are compared in accordance with the approach of Bland-Altman giving mean differences (\u0394), limits of agreement (LoA) and two regression coefficients (Intercept / Slope and Pearson\u2019s correlation coefficient).(PDF)Click here for additional data file.S10 TableAir trapping (E/I MLA) calculated for fully automatic and manually corrected segmentation on B60f scans at baseline, 3, 12 and 24 months. The last column summarizes all time points. All values are separately calculated for the right upper (RUL), middle (RML) and lower lobe (RLL), the left upper lobe (LUL), the lingula (LLi), the left lower lobe (LLL), and also combining left upper lobe and lingula into one lobe (LUL+LLi). Both methods are compared in accordance with the approach of Bland-Altman giving mean differences (\u0394), limits of agreement (LoA) and two regression coefficients (Intercept / Slope and Pearson\u2019s correlation coefficient).(PDF)Click here for additional data file."} +{"text": "Lias). These models have been mated with Ins2Akita/+ mice, a type I diabetic mouse model. We compare the major pathologic changes and oxidative stress status in two new strains of the mice with controls. Our results show that Ins2Akita/+ mice with under-expressed Lias gene, exhibit higher oxidative stress and more severe DN features . In contrast, Ins2Akita/+ mice with highly-expressed Lias gene display lower oxidative stress and less DN pathologic changes. Our study demonstrates that strengthening endogenous antioxidant capacity could be an effective strategy for prevention and treatment of DN.Oxidative stress is implicated in the pathogenesis of diabetic nephropathy (DN) but outcomes of many clinical trials are controversial. To define the role of antioxidants in kidney protection during the development of diabetic nephropathy, we have generated a novel genetic antioxidant mouse model with over- or under-expression of lipoic acid synthase gene ( Lias gene leads to mouse embryonic death, further underscoring the pivotal role LA plays in antioxidant defense and as a metabolic requirement [Alpha-lipoid acid is a potent antioxidant produced in mitochondria by lipoic acid synthase (LIAS) . Completuirement .Ins2Akita/+ type 1 diabetic mice with different levels of endogenous Lias gene expression, we sought to define the role of oxidative stress in the onset and development of DN and obtain a better understanding of impact of antioxidants.Diabetic nephropathy (DN) is a leading cause of end-stage renal disease . OxidatiLias gene and was replaced after recombination with a cassette. The cassette consisted of the 3\u2019-UTR sequences of bovine growth hormone gene (bGH) and a Neo gene, two lox P sites flanking the two fragments, and followed by the 3\u2019-UTR of cFos gene. Colonies surviving after selection with G418 and ganciclovir were first screened by PCR with the following primers: a common primer (5\u2032-CTA AAG TGT AGC CAA GCC CT-3\u2032), a primer for screening Lias 3\u2019-UTR (5\u2032-CCT CCT CAG CTA CTG ACA TT-3\u2032), a primer for bGH 3\u2019-UTR (5\u2032-GAG GCA AAC AAC AGA TGG CT-3\u2019) and a primer for cFos 3\u2019-UTR (5\u2032-CTT CTC TGA CTG CAG ATC CT-3\u2019). Targeted Embryonic stem (ES) cells were identified by the presence of approximately 200 bp PCR product for bGH 3\u2019-UTR, and 300 bp after Cre recombinase-mediated recombination. Germline recombination was achieved using the B6.FVB-Tg (EIIa-Cre) stock (JAX#3724). These results were confirmed by Southern blot analysis.The targeting construct prior to recombination consisted of a 3\u2019-UTR of the endogenous LiasLow/Low) and hypermorphic (LiasHigh/High) Lias mice in C57BL/6 genetic background, with 25% or 150% of wildtype Lias gene expression respectively, were mated with C57BL/6-Ins2Akita/+ diabetic mice (JAX#3548), an established mouse model of type I diabetes mellitus [Ins2Akita/+ mice have a mutation changing cysteine 96 to tyrosine in the insulin 2 gene and exhibit marked hyperglycemia as early as 4 weeks of age [LiasLow/LowIns2Akita/+ males and 9 B6-LiasHigh/HighIns2Akita/+ males were obtained from crossing C57BL/6-Ins2Akita/+ female mice and LiasLow/Low or LiasHigh/High male mice. Only males were phenotyped in this study because Ins2Akita/+ females on the B6 genetic background displayed much less severe diabetic phenotype than the males [Lias+/+Ins2Akita/+ mice served as a control. In addition, the mice were fed normal mouse chow and had ad libitum access to autoclaved water. All animal protocols were approved by the University of North Carolina at Chapel Hill Institutional Animal Care and Use Committee (Protocol numbers: 13-208-0 and16-153-0).The hypomorphic on samples obtained after a 5-hour fasting period. Plasma glucose, total cholesterol and triglyceride were examined using assay kits . Lactic acid concentration in tissues was determined as described . PyruvatLiasHigh/HighIns2Akita/+ and LiasLow/LowIns2Akita/+ mice was assessed by ELISA using Albuwell according to the manufacturer\u2019s instructions . Urinary creatinine levels were determined by the Creatinine Kit (Exocell) according to the manufacturer\u2019s instructions.At 28 weeks of age, individual mice were placed in metabolic cages to record food consumption, water intake, body weight, and urine output for 48 hours prior to sacrifice. Urine albumin in Urinary monocyte chemoattractant protein-1 (MCP-1) excretion was measured as markers of renal inflammation. The MCP-1 concentration was measured using an ELISA assay kit according to the manufacturer\u2019s instructions. The ELISA kit was specific for mouse MCP-1 and sensitive down to 2 pg/ml. The MCP-1 concentration was normalized to the urinary creatinine concentration.Systolic blood pressure (BP) was determined in conscious mice using a tail-cuff method . The firMice were perfused at 120 mmHg with 0.9% saline containing heparin, followed by 4% paraformaldehyde and then embedded in paraffin and 3-\u03bcm sections were cut. Sections were stained with hematoxylin and eosin (H&E) and Periodic acid-Schiff's base (PAS) for examination by light microscopy. Mesangial matrix expansion (MME) was examined in a blinded fashion and scored from 0 to 4 according to the ratio of glomerular expansion area/normal area: score 0, a normal glomerulus; score 1, increased mesangial matrix <25% of glomerular tuft; score 2, MME of 25%\u201350% of glomerular tuft; score 3, MME of 50%\u201375%; and score 4, MME of >75% of the tuft. MME was derived from assessment of three glomerular profiles on each mouse. Next, the kidney cortex was conventionally prepared for transmission electron microscopy. The samples were examined with an electron microscope . Three to four kidney samples from each experimental group were randomly chosen for electron microscopic observation. Thickness of glomerular basement membranes (GBM) was measured from transmission electron microscopy photos at 5 specific points along each of 10 randomly selected glomerular capillaries to determine an average GBM thickness for 5 mice in each group. Mitochondrial damage in proximal tubules was examined in each genotypic group. Briefly, under the same magnification , about 200\u2013300 mitochondria from 10\u201312 randomly selected fields in each genotypic group were observed. The Degree of the mitochondrial damage was assessed according to the following scale from 0 to 3: 0) indicating a normal structure, 1) normal with slight swelling, 2) mitochondrial swelling and cristae dilated/disorder, and 3) mitochondrial vacuolization. On the basis of the above criteria, the degree of mitochondrial damage in the different groups of mice were scored and the total scores of mitochondria per group were summarized and then divided by total counted mitochondrion number in each group to get the ratio of damaged over total counted mitochondrion number. The ratio indicates the degree of mitochondrial damage.To evaluate oxidative stress, the concentration of urinary 8-isoprostane was measured using an enzyme immunoassay and expressed relative to the level of urine creatinine following the manufacturer\u2019s protocol . Systemic oxidative stress in blood was determined using 4-Hydroxynonenal (4-HNE) assay kit in accordance with manufacturer\u2019s specifications .Total protein was extracted from the renal cortical tissues with RIPA buffer and protein concentration was determined by BCA protein assay method following manufacturer\u2019s instructions. Western blot analysis was performed using a rabbit polyclonal antibody against mouse LIAS , and (voltage-dependent anion-selective channel protein 1) VDAC1 as mitochondrial loading control and the protein bands were quantified with Image Quant LAS4000 software .\u03b2-actin as the reference gene in each reaction [Lias, superoxide dismutase 2 (Sod2), transforming growth factor \u03b21 (Tgf\u03b21), nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and NADPH oxidase 4 (Nox4) were examined.Total RNA was extracted from kidney cortex using an ABI 6700 Automated Nucleic Acid Workstation following the manufacturer\u2019s protocol . Relative mRNA amounts were determined using real-time quantitative reverse transcriptase-PCR (Applied Biosystems) with reaction . The expP<0.05 was considered significant. P values are obtained for comparisons among LiasLow/LowIns2Akita//+, LiasHigh/HighIns2Akita/+ and Lias+/+Ins2Akita/+ mice using one-way or two-way ANOVA. Post hoc pairwise comparisons were performed by Tukey\u2013Kramer honestly significant differences (HSD) test .Data were expressed as mean \u00b1 standard error of the mean (SEM). Lias mice with over- or under-expression of the Lias gene was based on genetic modification of the 3\u2019-UTR . ES cell lines with a correctly targeted 3\u2019-UTR were identified and injected into C57BL/6J recipient blastocysts, which were transferred to the uteri of CD-1 pseudopregnant dams. These mice initially produced stabilized transcripts of the Lias gene using the 3\u2019-UTR sequence of bGH, but were changed to unstable transcripts using the 3\u2019-UTR from the cFos gene after Cre-LoxP recombination was induced. The LiasHigh/+ mice were crossed with Tg (Ella-cre) mice that expressed Cre-recombinase in testis and thus the homozygous offspring (LiasLow/Low) generated from these LiasLow/+ founder males expressed low levels of Lias. Since plasma and tissue LA were not directly measured due to a technical difficulty [Lias gene expression using RT-PCR and hypermorphic (LiasHigh/High) Lias mice, with 25% or 150% of wildtype Lias gene expression, respectively, were mated with Ins2Akita/+ diabetic mice, an established mouse model of diabetes mellitus that can mimic early stages of DN [LiasHigh/HighIns2Akita/+ and LiasLow/LowIns2Akita/+ mice exhibited the different diabetic phenotypes. LiasLow/LowIns2Akita/+ mice had significantly lower body weight compared to both LiasHigh/HighIns2Akita/+ and Lias+/+Ins2Akita/+ mice as shown in P<0.05). Both LiasHigh/HighIns2Akita/+ and LiasLow/LowIns2Akita//+ mice manifested hyperglycemia at 28 weeks of age but there was no significant difference between the two groups of mice. In addition, plasma total cholesterol and triglyceride levels were similar among LiasHigh/HighIns2Akita/+, LiasLow/LowIns2Akita/+, and Lias+/+Ins2Akita/+ mice in LiasLow/LowIns2Akita/+ mice as compared with LiasHigh/HighIns2Akita/+ and Lias+/+Ins2Akita/+ mice of TGF-\u03b21.Consistent with Western blot results for LIAS protein, RT-PCR analysis of kidney cortex showed significantly increased s of age . Interesa/+ mice . To idenoic acid . In addiita mice suggestsNrf2 gene expression in 28-week-old LiasLow/LowIns2Akita/+ and LiasHigh/HighIns2Akita/+ mouse kidney. Unlike Sod2, Nox4 and Tgfb1 transcripts, no significant changes in Nrf2 expression were observed in response to Lias gene manipulation (NRF2 is a key transcription factor for regulation of antioxidant defense and LA hpulation .LiasLow/LowIns2Akita/+ mice compared with Lias+/+Ins2Akita/+ and LiasHigh/HighIns2Akita/+ mice (Inflammation is thought to be a pathogenic factor in the initiation of DN and monocyte chemoattractant protein-1 (MCP-1) is considered as a major mediator of inflammation in DN patients. To further probe the underlying mechanism(s) of endogenous oxidant injury in relation to inflammation in diabetes mellitus, we measured urinary MCP-1. Our results showed that there were significantly increased urinary MCP-1 levels in a/+ mice .Lias+/-Ins2Akita/+ mice with an approximately 50% reduced Lias gene expression, and the aforementioned pathology was associated with significantly enhanced oxidative stress [Lias transcript stability to generate two lines of novel antioxidant mouse models. In one, Lias gene expression was reduced to roughly 25% of normal, a level sufficient to maintain viability . In the other, Lias gene expression was increased to approximately 150% of normal expression level, which is anticipated to afford better protection, than wildtype mice, against oxidative stress. When combined with the diabetogenic Ins2Akita mutation, the double mutants allow evaluation of the impact of two different antioxidant baselines (Low or High) on the development of DN. As hypothesized, the new diabetic models exhibited divergent renal responses to diabetogenic stress when compared with Lias+/+Ins2Akita/+ mice. Major DN pathological changes such as albuminuria and mesangial expansion in LiasLow/LowIns2Akita/+ mice were significantly worse than those in Lias+/+Ins2Akita/+ mice whereas the two pathological changes were less severe in LiasHigh/HighIns2Akita/+ mice. The data further support the correlation between Lias transcript abundance and inferred endogenous LA concentration; levels of two reliable common oxidative stress biomarkers, plasma 4-NHE and urine 8-isoprostane, were both significantly higher in LiasLow/LowIns2Akita/+ mice and significantly lower in LiasHigh/HighIns2Akita/+ mice compared to Lias+/+Ins2Akita/+ controls. These results are consistent with our previous data where we used different lipid peroxidation markers, like TBARS and, antioxidant marker such as ratio of GSH/GSSG to reveal significantly increased oxidative stress and decreased endogenous antioxidant capacity in Lias+/-Ins2Akita/+ mice. Since lipoic acid plays a central role in the antioxidant network, the significantly increased oxidative stress and decreased endogenous antioxidant capacity observed in Lias+/-Ins2Akita/+ mice were very likely due to the impairment of the antioxidant defense system. In addition, our data demonstrate that Lias overexpression effectively attenuates albuminuria and kidney disorders without exerting a significant hypoglycemic effect in LiasHigh/HighIns2Akita/+ mice, suggesting that the protective effect of LA results primarily from its antioxidant capacity rather than from a direct hypoglycemic effect. These findings confirm that LIAS-generated lipoic acid plays a vital role in the early development of DN by demonstrating that diabetic mice with low endogenous antioxidant capacity manifestly increased ROS-mediated renal stress. In particular, the new models provide very strong proof-of-principle that an increased antioxidant reservoir could represent a powerful new tool in the prevention and / or treatment of diabetic kidney disease. A novel finding obtained from our current study underlines the importance of antioxidants for mitochondrial protection in the retardation of DN. Growing evidence indicates that mitochondria play a critical role in the initiation and development of DN. Mitochondria are believed to be the major organelles involved in superoxide generation. They consume approximately 85% of the oxygen used by cells and overproduction of superoxide anions may occur by excessive electron leak in the mitochondrial electron transport chain during diabetes [Ins2Akita/+ mice with 50% of reduced Lias gene expression than Lias+/+Ins2Akita/+ littermate controls. In the current studies, we consolidate our previous observation that mitochondrial damage in proximal tubules is a predominant pathological feature of DN by demonstrating that reduction in Lias gene expression in LiasLow/LowIns2Akita/+ mice and increased Lias expression in LiasHigh/HighIns2Akita/+ mice impact mitochondrial integrity and function. Through use of in vitro LIAS knockdown studies it has been reported that ROS may decrease the mitochondrial membrane potential [Ins2Akita/+ mice specifically overexpressing catalase, a key antioxidant enzyme in renal proximal tubular cells, had reduced renal oxidative stress and attenuated progression of DN without changing blood glucose concentration [Ins2Akita/+ derived \u03b2-cells have increased mitochondrial dysfunction, oxidative stress, mitochondrial DNA damage, and alterations in mitochondrial protein levels that contribute to \u03b2-cell dysfunction [In our previous study, the pathologic changes of diabetic nephropathy (DN) were exacerbated in e stress . To veridiabetes . On the diabetes . Our preotential . Given totential , we presfunction .Lias+/-Ins2Akita/+ mice showed that systemic and urinary oxidative stress markers significantly increased, whereas endogenous antioxidant capacity significantly decreased, when Lias gene expression levels was approximately 50%. Furthermore, reduced Lias gene expression in Lias+/-Ins2Akita/+ mice was associated with more severe DN pathological changes compared with Lias+/+Ins2Akita/+ mice. In particular, a large number of damaged mitochondria were detected in mouse proximal tubule epithelial cells as a unique phenomenon. The results observed in LiasLow/LowIns2Akita/+ mice have confirmed that reduced Lias gene expression leads to decreased endogenous antioxidant capacity and mitochondrial damage in the proximal tubules. On the other hand, in LiasHigh/HighIns2Akita/+ mice harboring increased Lias gene expression, endogenous oxidant capacity and mitochondrial integrity are protected. Our data also indicate that kidney proximal tubules can serve as a window via which alternation of mitochondrial status due to oxidative stress can be assessed in diabetic nephropathy.LIAS synthesizes lipoic acid in mitochondria. Lipoic acid has a high reductive capacity and actively participates in the recycling of vitamin C and E. Diabetes mellitus is characterized by increased oxidative stress that negatively impacts mitochondrial integrity and function. Hence the physiologic importance of LIAS in diabetes is through its role in lipoic acid synthesis; lipoic acid may play a vital role in mitochondrial protection from oxidative stress and thus maintain energy balance during diabetes. Our previous data obtained from in Lias expression and LA levels under diabetic conditions:Based on ours and, other investigator\u2019s data , we propLias gene further highlights this relationship between endogenous antioxidant levels, ROS concentrations and DN pathologic changes. That is, low endogenous antioxidant capacity will result in high levels of ROS and more severe DN.Excess ROS generated in diabetic mellitus impairs the antioxidant defense system, indicated by decline of reduced GSH levels, leading to further accumulation of ROS. The latter is reflected by a significantly enhanced lipid peroxidation levels in body and urine. Our new animal model with low expression of LiasLow/LowIns2Akita/+ mice could damage mitochondria. We found that significant mitochondrial damage in LiasLow/LowIns2Akita/+ mice; other investigators have also demonstrated decreased mitochondrial membrane potential using in vitro LASY knockdown method [Excessive ROS in diabetes mellitus damages mitochondria. Mitochondria are a major site of ROS generation and are vulnerable targets for ROS; hence, accumulated ROS due to insufficient LA protection in n method .Sod2 (a major antioxidant enzyme which responds to enhanced superoxide production in mitochondria) markedly increases in kidney cortex of LiasLow/LowIns2Akita/+ mice; the latter observation has been previously reported in the Lias+/-Ins2Akita/+ diabetic mice with 50% reduced LIAS [LiasLow/LowIns2Akita/+ mice with increased ROS is not surprising.To investigate the mechanisms through which lipoic acid protects mitochondria in kidney proximal tubules, we examined gene expression of several common antioxidant enzymes in kidney cortex, including Sod series and glutathione peroxidase. Our data, using these novel mouse models, identify Sod2 as an antioxidant target by showing that expression of ced LIAS . SOD2 coNox4 gene, a biomarker of oxidative stress in the diabetic mellitus, has particularly high expression in the kidney [Nox4 gene expression was significantly increased in LiasLow/LowIns2Akita/+ and decreased in LiasHigh/HighIns2Akita/+ kidney cortex, respectively, suggesting that NADPH oxidase 4 is an antioxidant target of lipoic acid.We attempted to identify antioxidant targets of LA in the diabetic kidney cortex. In addition to dysfunctional mitochondria, one of the most prominent sources of ROS is from the NADPH oxidase (NOX) activity \u201329. Amone kidney . Our resNrf2 gene expression in 28-week-old LiasLow/LowIns2Akita/+ and LiasHigh/HighIns2Akita/+ mouse kidney. Hence, the renal protective action mediated by increased LIAS-generated LA is not accompanied by Nrf2 transcription alternation.NRF2 is a key transcription factor for regulation of antioxidant defense . AlthougTgf\u03b21 gene expression in LiasHigh/HighIns2Akita mice may play a role in attenuation of the adverse effect of diabetes mellitus on kidney structure and function. In addition, our results obtained from previous and the current studies using different degrees of Lias deficiency mouse models have clearly demonstrated that renal proximal tubules, with numerous mitochondria required for their important absorption functions, are sensitive to mitochondrion damage and that they are an ideal region of the nephron for assessment of mitochondria status during development of diabetic nephropathy. It is also worth studying the impact of mitochondrial damage on resorption in order to ascertain which mitochondrial component(s) is/are injured and the mechanism through which the deficiency of endogenous antioxidant protection develops.Several studies have revealed increased expression of TGF-\u03b21 in renal glomeruli in experimental models of diabetes . TGF-\u03b21 LiasLow/LowIns2Akita/+ mice compared with Lias+/+Ins2Akita/+ and LiasHigh/HighIns2Akita/+ mice. This observation indicates that increased inflammation occurs in LiasLow/LowIns2Akita/+ mice. The increased MCP-1 is likely mediated by NF-kB in response to enhanced kidney oxidative stress [Accumulating evidence support a role for inflammation in DN. In particular, the inflammatory cells infiltrate and significantly higher levels of cytokines (chemokines) including monocyte chemoattractant protein-1 (MCP-1), accompanies DN . MCP-1 ie stress , 42. ThiLias+/-Ins2Akita/+ mice are significantly greater than those in Lias+/+Ins2Akita/+ mice [Lias gene expression in the different genetic background could exhibit different traits.In a previous study, we showed that plasma glucose levels in a/+ mice . Howevera/+ mice , 44. We LiasHigh/HighIns2Akita/+ and LiasLow/LowIns2Akita/+ mice manifest variations in levels of endogenous antioxidant capacity in kidneys that lead to different degrees of diabetic pathologic changes. The results have clarified the role of antioxidants in the early development of diabetic nephropathy and strongly suggest that protection of mitochondria is a novel therapeutic target for effective antioxidant therapy of DN.In summary, our data clearly indicate that We conclude that these new antioxidant mouse models are suitable to elucidate the contributions of oxidative stress in the pathogenesis of diabetic kidney disease."} +{"text": "Scientific Reports6: Article number: 31281; 10.1038/srep31281 published online: 08092016; updated: 09162016.The original version of the Article contained typographical errors.In the Abstract,\u201cA decline in the number of males born relative to the number of females was also observed. ECs, including diethylhexyl phthalate (DEHP) and polychlorinated bisphenol 153 (PCB153), were detected in adult dog testes and commercial dog foods at concentrations reported to perturb reproductive function in other species\u201d.now reads:\u201cA decline in the number of males born relative to the number of females was also observed. ECs, including diethylhexyl phthalate (DEHP) and polychlorinated biphenyl 153 (PCB153), were detected in adult dog testes and commercial dog foods at concentrations reported to perturb reproductive function in other species\u201d.In the Results section under subheading \u2018Establishing concentrations of environmental chemicals in canine adult testis and semen\u2019,\u201cSeven polychlorinated bisphenol (PCB) congeners, 5 polybrominated diphenyl ether (PBDE) congeners and diethylhexyl phthalate (DEHP) were detected in testis \u201d.now reads:\u201cSeven polychlorinated biphenyl (PCB) congeners, 5 polybrominated diphenyl ether (PBDE) congeners and diethylhexyl phthalate (DEHP) were detected in testis \u201d.These errors have now been corrected in the PDF and HTML versions of the Article."} +{"text": "The aim of this article is to assess HFE C282Y gene mutations as a predictor of sustained virological response (SVR) to anti-hepatitis C virus (HCV) treatment in Egyptian patients.One hundred and forty chronic hepatitis C (CHC) patients were divided into two groups: 70 patients achieved SVR and 70 patients were nonresponders (NRs). All patients were subjected to quantitative polymerase chain reaction (PCR) at baseline, 12 and 24 weeks after therapy commencement. Deoxyribonucleic acid (DNA) sequencing for HFE (C282Y) was done by restriction fragment length polymorphism PCR.Sixty five patients did not have mutation and 5 patients had C282Y mutation (GA) with SVR. While 45 NRs had heterozygous C282Y mutation (GA), 4 patients (5.7%) had homozygous mutation (AA) and 21 patients (30%) had no mutation (GG). The parameters of elevated iron were significantly associated with C282Y mutation. However, there was no significant difference regarding ferritin values and C282Y mutation in NR patients.Iron overload was frequently detected in CHC patients and associated with C282Y mutation, while biochemical markers of iron overload and C282Y HFE mutation were negative prognostic factor.How to cite this article: Mehrez MI, Fattah DSA, Azeem NAA, Saleh MA, Mostafa KM. Hemochromatosis Gene Polymorphism as a Predictor of Sustained Virological Response to Antiviral Treatment in Egyptian Chronic Hepatitis C Patients. Euroasian J Hepato-Gastroenterol 2017;7(2):154-157. Due to these effects, iron can be considered a proinflammatory, profibrogenic factor, and a potential carcinogen. Since the implementation of serological diagnostic tests for HCV identification, elevated serum iron overload indices or appearance of iron deposits in liver cells have been observed in 10 to 40% of patients with CHC and 50% of patients suffering both from CHC and hepatocellular carcinoma.4 Some investigations have shown an association between elevated serum iron indices or high hepatic iron concentration (HIC) and the lack of SVR in CHC patients,5 whereas others have shown that there is no positive correlation between HIC and decreased frequency of SVR.6 In 2006, Bonkovsky et al7 found the presence of iron in endothelial cells with triad iron score as a predictor of decreased SVR. These contradictory results from different parts of the world may possibly have their source in ethnic differences and the variable polymorphisms of iron metabolism-related genes found in different populations. The aim of this work is to assess the value of HFE C282Y gene mutations as a predictor of SVR to antiviral treatment in Egyptian patients with CHC virus infection.The World Health Organization has declared hepatitis C as a global health problem, with approximately 3% of the world\u2019s population infected with HCV.The study was conducted on 140 CHC patients (based on the presence of persistently elevated liver enzymes for at least 6 months and detection of HCV ribonucleic acid by PCR technique) who took antiviral treatment. The patients were divided into two groups: Group I consisted of 70 patients who achieved SVR after antiviral treatment, and group II consisted of 70 patients who did not respond to antiviral treatment (NR). Patients with malignancy, decompensated cirrhosis, hepatitis B virus coinfection, or other causes of liver disease were excluded. All patients were subjected to full history taking, clinical examination, laboratory investigations including liver and kidney biochemical profile, alfa fetoprotein (AFP), viral load, and specific tests of our study: Complete iron profile, molecular study for HFE (C282Y).The data about patient profiles are shown in 8 The mechanism by which iron accumulates in liver infected with chronic HCV has not yet been established. Serum iron and ferritin levels were increased in patients with CHC because of their release from hepatocellular stores in association with cell necrosis.9 Individuals with serum iron levels in the upper range of normal as a result of genetic polymorphisms or a high iron diet may be predisposed to develop more severe chronic HCV infections.9 Several studies10 have found that heterozygous C282Y mutations are associated with hepatic iron loading in CHC patients. Iron overload seems to impair antigen-specific immune responses by decreasing the generation of T cells and by impairment of natural killer and T helper cell function. Piperno et al11 suggested that iron overload in patients with hemochromatosis may contribute to the persistence of HCV infection, and iron overload may in theory promote viral replication. The amount of hepatic iron has been identified as one of these factors that adversely affect the likelihood of response to interferon-alfa; those patients with higher hepatic iron content are less likely to respond to interferon therapy.12 In our study, there was a correlation between HFE gene mutation and iron overload. We considered transferrin saturation index (TSI) as the most specific and sensitive parameter in identifying iron overload as it showed a significant statistical difference between responder group (27.5%) and NR (34.5%) group with p-value <0.001. But there was no significant difference for serum ferritin, S iron (p = 0.02), TIBC (p > 0.001) transferrin (p = 0.016), sTfR (p > 0.001), but our study provides evidence supporting that the HFE gene mutations are associated with significant abnormalities of iron metabolism and suggests that patients with CHC accumulate iron as a result of interplay between genetic and acquired factors. We noticed that A allele is associated with higher iron parameters and lower TIBC and the homozygous mutation (AA) is associated with higher iron indices. The wildtype (GG) is lower than the heterozygous mutation (GA) genotype. There is a statistically significant difference between gene polymorphism and iron parameters with p-value <0.001 for each of S. iron, TIBC, TS%, and S. ferritin, with p-value 0.033 as regarding transferrin and by 0.026 as regarding sTfR. Sustained virological response rates were lower among patients with HFE gene mutations compared with those with HFE gene wildtype. In our study, 54 of 140 (38.5%) patients have mutation [50 heterozygous (GA) and 4 homozygous (AA)] and 86 have no mutation (wild-type GG). All homo and 45 from heterozygous mutation did not respond to treatment; 92% (92.9%) of the SVR group have no (GG) mutation and 7% carry C282Y mutation (GA), while 64.3% of NR group carry heterozygous C282Y mutation (GA), 5.7% carry homozygous mutation (AA), and 30% are without mutation (GG). Therefore, HFE gene mutations may act synergically with CHC in the development of liver damage, predicting a higher rate of nonresponse to therapy. Our results correlate with those of Sini et al,13 who stated that 69 CHC patients with end-of-treatment response were lower among patients with HFE gene mutations compared with those with HFE gene wildtype (p = 0.005) and TSI showed a significant statistical difference between HFE mutant patients (50%) and wild-type homozygotes (43.4%) (p < 0.01). Coelho-Borges et al14 had similar results in 2002 when they studied 44 Brazilian patients. They showed that SVR was achieved in 0 of 16 patients with HFE gene mutations and 11 (41%) of 27 patients without HFE gene mutations (p = 0.002). They concluded that heterozygosity for H63D and/ or C282Y HFE gene mutation predicted absence of SVR to combination treatment with interferon and ribavirin in patients with CHC, non-1 genotype and serum ferritin levels above 500 ng/mL. Our results did not correlate with those of Li et al,15 who showed that H63D mutation was associated with a significantly higher SVR rate , while the C282Y mutation was not . We do not agree with Lebray et al16 who based on a large cohort of HCV-infected patients found an opposite effect of iron blood parameters and the H63D mutation on the antiviral efficacy of interferon-alfa used alone or in combination therapy with exception of six C282Y heterozygote patients that displayed no sustained response; but this group was too small to allow the detection of a significant difference with any other group. Increased iron stores may affect the course of viral infection in various ways: First, increased HIC may facilitate viral replication and in vitro data suggest that iron facilitates HCV replication in cultured hepatocytes.17 Second, iron loading was demonstrated to enhance HCV pathogenicity.18Chronic hepatitis C patients have frequently elevated serum iron stores and elevated HIC, which has been associated with a poor response to interferon-alfa."} +{"text": "Listeria monocytogenes and Salmonella typhimurium. In response to both bacteria we identified thousands of genes that significantly change isoform usage in response to infection, characterized by an overall increase in isoform diversity after infection. In response to both bacteria, we found global shifts towards (i) the inclusion of cassette exons and (ii) shorter 3\u2019 UTRs, with near-universal shifts towards usage of more upstream polyadenylation sites. Using complementary data collected in non-human primates, we show that these features are evolutionarily conserved among primates. Following infection, we identify candidate RNA processing factors whose expression is associated with individual-specific variation in isoform abundance. Finally, by profiling microRNA levels, we show that 3\u2019 UTRs with reduced abundance after infection are significantly enriched for target sites for particular miRNAs. These results suggest that the pervasive usage of shorter 3\u2019 UTRs is a mechanism for particular genes to evade repression by immune-activated miRNAs. Collectively, our results suggest that dynamic changes in RNA processing may play key roles in the regulation of innate immune responses.The contribution of pre-mRNA processing mechanisms to the regulation of immune responses remains poorly studied despite emerging examples of their role as regulators of immune defenses. We sought to investigate the role of mRNA processing in the cellular responses of human macrophages to live bacterial infections. Here, we used mRNA sequencing to quantify gene expression and isoform abundances in primary macrophages from 60 individuals, before and after infection with Changes in gene regulation have long been known to play important roles in both innate and adaptive immune responses. While transcriptional responses to infection have been well-characterized, much less is known about the extent to which co-transcriptional mechanisms of mRNA processing are involved in the regulation of immune defenses. In this study, we sought to investigate the role of mRNA processing in the cellular responses of human macrophages to live bacterial infection. Using primary human macrophages derived from whole blood samples from 60 individuals, we sequenced mRNA both before and after infection with two live bacteria. We show that immune responses to infection are accompanied by pervasive changes in mRNA isoform usage, with systematic shifts towards increased cassette exon inclusion and shortening of Tandem 3\u2019 UTRs post-infection. These patterns are conserved in nonhuman primates, supporting their functional importance across evolutionary time. Complementary microRNA profiling revealed that shortened 3\u2019 UTRs are enriched for target sites of macrophage-expressed miRNAs, many of which are specifically activated after infection to regulate the innate immune response. Our results therefore provide the first genome-wide empirical support for the idea that actively regulated shifts towards shorter 3\u2019 UTRs might allow specific genes to evade repression by immune-activated miRNAs. Innate immune responses depend on robust and coordinated gene expression programs involving the transcriptional regulation of thousands of genes \u20133. TheseAlthough much attention has been devoted to characterizing transcriptional changes in response to infectious agents or other immune stimuli, we still know remarkably little about the contribution of co-transcriptional changes\u2013specifically, changes in alternative pre-mRNA processing\u2013to the regulation of the immune system \u20138. AlterListeria monocytogenes or Salmonella typhimurium in primary human macrophages. Because of the distinct molecular composition of these two pathogens and the way they interact with host cells, they activate distinct innate immune pathways after infection [46 [4629] ,29. ThisIRF5 transcription factor [MAPKAP1 and MAP2K4 [Specifically, our data support previous hypotheses and find evidence for that 3\u2019 UTR shortening in macrophages is associated with the loss of target sites for a subset of immune-regulated miRNAs, including miR-146b, miR-125a, and miR-151b . These mn factor and the d MAP2K4 \u2013all of wWhile our results point to the importance of 3\u2019 UTR shortening to allow genes to evade post-transcriptional regulation by miRNAs, this is not generalizable to all miRNAs induced in response to infection. Indeed, some of the miRNAs most highly induced after infection, such as miR-155 and miR-29b, are not enriched for binding sites in the longer 3\u2019UTR regions. Thus, while miRNA regulation is crucial to mammalian immune response \u201362, the trans to drive global shifts towards inclusion of cassette exons or usage of upstream proximal polyadenyalation sites. Taking advantage of our relatively large sample size, we were able to identify candidate trans-factors whose up-regulation upon infection might have widespread impacts on RNA processing patterns, including some known regulators of alternative splicing or 3\u2019end processing , combined with no changes in the levels of U1 snRNA , 6 common chimpanzees, , and 6 rhesus macaques . Human samples were acquired with informed consent and ethics approval from the Research Ethics Board . Non-human primate samples were acquired in accordance with individual institutional IACUCC requirements. For all species, 1ml of blood was drawn into a media-containing tube spiked with ultrapure LPS or endotoxin-free water (control). Samples were stimulated with 1ug/ml LPS, at 37C for 4 hours before total blood leukocytes were isolated and RNA collected. Genome-wide gene expression profiles of untreated and infected/treated samples were obtained by RNA-sequencing for both mRNA transcripts and small RNAs. After a series of quality checks , mRNA transcript abundances were estimated using RSEM [sh was applied to measure the diversity of isoforms for each target gene before and after infection. Changes across individual RNA processing events were quantified using the MISO software package (v0.4.9) [Complete details of the experimental and statistical procedures can be found in SI Materials and Methods. Briefly, blood samples from 60 healthy donors were obtained from Indiana Blood Center with informed consent and ethics approval from the Research Ethics Board at the CHU Sainte-Justine (protocol #4022). All individuals recruited in this study were healthy males between the ages of 18 and 55 y old. Blood mononuclear cells from each donor were isolated by Ficoll-Paque centrifugation and blood monocytes were purified from peripheral blood mononuclear cells (PBMCs) by positive selection with magnetic CD14 MicroBeads (Miltenyi Biotec). In order to derive macrophages, monocytes were then cultured for 7 days in RPMI-1640 (Fisher) supplemented with 10% heat-inactivated FBS , L-glutamine (Fisher) and M-CSF . After validating the differentiation/activation status of the monocyte-derived macrophages we infected them at a multiplicity of infection (MOI) of 10:1 for ing RSEM and Kalling RSEM and micring RSEM . To dete(v0.4.9) using de(v0.4.9) . We used(v0.4.9) .Institutional ethics (IRB) approval was obtained by the CHU Sainte-Justine Institutional Review Board (approved project #3557) and all subjects gave written informed consent prior to participation.http://www.ncbi.hlm.nih.gov/geo/) under the accession number GSE73765, which comprises of the mRNA-seq data (GSE73502) and small RNA-seq data (GSE73478).Data generated in this study have been submitted to the NCBI Gene Expression Omnibus Click here for additional data file.S1 Fig(A) Volcano plots of differential expressionafter infection with Listeria and Salmonella in the left and right panels, respectively.\u2013log10 p-values (y-axis) testing for differential expression are plotted against average log2 fold changes in expression levels (x-axis) for genes that are not differentially expressed (black) and genes that with significant differential expression after infection (FDR \u2264 0.1% and |log2(fold change)| \u2265 0.5; blue). (B) Distribution of p-values for the differential isoform usage test upon infection with Listeria and Salmonella. Expected distribution of p-values under the null hypothesis of no significant difference between the mean relative isoform usage is shown in grey. (C) A comparison of DIU effect sizes between DIU genes at 1% FDR and the background of all genes (lighter colors). Effect sizes are defined as maximum change in relative isoform abundances per gene upon infection. (D) Distributions of \u0394Shannon diversity index (\u0394SDI) after infection with Listeria and Salmonella. Null distribution (black dotted line) was generated by permuting samples across conditions. (E) The percentage of genes with a dominant isoform before infection (left) and the fraction of these genes where the dominant isoform changes after infection (right).(PDF)Click here for additional data file.S2 Figgrey, while the predominant isoform in infected samples is in a color. Darker bars represent increased relative usage, while lighter bars represent decreased relative usage.Heatmaps below each example represent the variation in isoform usage across the 60 individuals, where each vertical bar represents one individual. The dominant isoform in non-infected samples is represented in (PDF)Click here for additional data file.S3 Fig(A) Proportion of genes with differential isoform usage (y-axis) among all genes (grey), differentially expressed genes that are down-regulated after infection (orange), and differentially expressed genes that are up-regulated after infection (green). (B)log2 fold changes (with standard error bars) for odds of low \u0394Shannon and high \u0394Shannon among the set of all the differentially expressed genes, up-regulated differentially expressed genes, and down-regulated differentially expressed genes, following infection with Listeria (dark pink) and Salmonella (gold).(PDF)Click here for additional data file.S4 Figx-axis) in \u0394\u03a8 values for each isoform in a given RNA processing category.Distribution of coefficient of variation ((PDF)Click here for additional data file.S5 FigPTK2B and STK40, that have significant changes in skipped exon usage both after 2 hours and 24 hours of infection with either bacteria.Sashimi plots for two genes, (PDF)Click here for additional data file.S6 Figy-axis) and TPMs from RNA-seq data (x-axis).Correlations between TPMs from 3' RNA-seq data ((PDF)Click here for additional data file.S7 Fig(A) Proportion of significantly changing events (x-axis) after 2 hours of infection (dark colors) and 24 hours of infection (light colors) with either Listeria (top) or Salmonella (bottom). Numbers indicate the significant events at corresponding timepoints using only the 6 individuals used for these cross-timepoint analyses. (B) Distribution of \u0394\u03a8 values (x-axis) for significantly changing events in each event type after either 2 hours (top) or 24 hours (bottom) of infection.(PDF)Click here for additional data file.S8 Fig(A) Distributions of overall fold changes in gene expression for genes that have significant splicing changes in each event type (colored boxplots) and genes with no splicing changes after infection (grey). Gene expression values are calculated using either full transcript models (top) or only constitutively included exons (bottom). (B) Distribution of spearman correlations between the \u0394\u03a8 of an event and the fold change in gene expression, across individuals, for events that are not significantly changing after infection (left) and significantly changing events (right). Solid lines represent the observed values and dotted lines represent a distribution of correlations after permuting the correspondence between \u0394\u03a8s and fold changes in gene expression.(PDF)Click here for additional data file.S9 Fig(A) Significantly enriched gene ontology categories for genes with significant skipped exon changes after infection. Color indicates the fold enrichment of the number of observed genes relative to the number of genes expected to be in that category. (B) Fold changes in gene expression after infection for 2 families of splicing factors (hnRNPs and SR proteins) relative to a background distribution of fold changes in all genes (C) Fold enrichments of splicing regulatory elements (SREs) in exonic regions and surrounding intronic regions (\u00b1100bp). SREs assessed included exonic splicing enhancers (green in exons), exonic splicing silencer (red in exons), intronic splicing enhancers (green in introns), and intronic splicing silencers (green in introns). Enrichments were calculated separately for significantly included skipped exons (top) and significantly excluded skipped exons (bottom). Shading of the bars indicates the significance of the enrichment.(PDF)Click here for additional data file.S10 Fig(A) Correlations between the mean \u0394\u03a8 values per individual for ALEs (x-axis) and TandemUTRs (y-axis). (B) Spearman correlations between the mean \u0394\u03a8 value per individual and the mean fold change of gene expression per individual for corresponding genes.(PDF)Click here for additional data file.S11 Fig(A) and Skipped exons (B), the top panel is a scatter plot of Spearman correlations between the individual-specific mean \u0394\u03a8 values and individual-specific fold change of gene expression values for all expressed genes (grey) in both Listeria (x-axis) and Salmonella (y-axis). Genes with significant correlations (FDR \u2264 1%) in both Listeria and Salmonella conditions are plotted in black, and those factors with known RNA-binding properties are colored by their functional category. The bottom panels show distributions of the average Spearman correlation values for each of the RNA-binding functional categories with significant correlations.For both Tandem UTRs (PDF)Click here for additional data file.S12 Figtop panel) and LCLs (bottom panel) in non-infected cells and in cells infected with Listeria or Salmonella for both 2 and 24 hours. BrdU incorporates into newly synthesized DNA and therefore the quantity of BrdU incorporated into cells (x-axis) is a direct indication of cell proliferation. No evidence for cellular proliferation was observed in macrophages, in contrast to the high rates of proliferating cells observed in our positive control population of LCLs.BrdU cell proliferation assay in macrophages ((PDF)Click here for additional data file.S13 FigListeria (pink) or Salmonella (yellow) infections.Barplots in grey indicate Tandem 3\u2019 UTRs that are not changing after infection, while colored barplots indicate Tandem 3\u2019 UTRs that are significantly changing after either (PDF)Click here for additional data file.S14 Figbrown) and extended (blue) regions of Tandem 3' UTRs that are either not changing or significantly changing after infection (left panel). While there 3' UTRs that are significantly changing generally have greater overall GC content, this is true for both the core and extended regions of the 3\u2019 UTRs. Thus, the distributions of relative GC content when comparing the regions is distributed around 1 for both significantly changing (yellow) and not changing (blue) Tandem 3' UTRs (right panel).The distribution of GC content in core ((PDF)Click here for additional data file.S15 Figgrey, n = 20), U1 , and total RNA concentrations after infection. 5s rRNA concentrations were calculated by taking the \u0394CT value from qPCR measurements across 20 samples. Relative U1 snRNA concentrations were calculated by taking the \u0394\u0394CT values from qPCR measurements across 20 samples, where U1 snRNA is measured relative to 5s rRNA concentrations in the same samples. Total RNA concentrations were estimated from Nanodrop measurements of RNA extraction yields across all 60 samples in our study. All samples (both non-infected and infected) were plated at exactly the same macrophage cellular density at the start of the experiment and we have confirmed that these macrophages do not proliferate either before or after infection . There are no significant shifts in the distribution of 5s rRNA or U1 snRNA fold changes after infection , while the distribution of total RNA fold changes are significantly increased after infection .Distributions of fold changes in 5s rRNA ((PDF)Click here for additional data file.S1 TableDetails of RNA-seq data for samples after infection at 2 hours or 24 hours. Libraries were made as described in Supplementary Section 1.2.1 in (XLSX)Click here for additional data file.S2 Tablelog2 fold change estimates, p-values for differential expression, and Benjamini-Hochberg adjusted p-values for differential expression between non-infected samples and both Listeria and Salmonella samples.Estimates of differential gene expression for the 14,954 genes tested by DESeq2, as described in Supplementary Section 1.3.2 in (XLSX)Click here for additional data file.S3 TableEnriched Gene Ontology categories for genes that are differentially expressed, or show differential isoform usage after infection. Gene Ontology analyses were performed using GSEA, as described in Supplementary Section 1.3.8 in (XLSX)Click here for additional data file.S4 TableEstimates of differential isoform usage for the 11,353 genes tested as described in Supplementary Section 1.3.3 in (XLSX)Click here for additional data file.S5 TableListeria and Salmonella, as described in Supplementary Section 1.3.4 in Estimates of Shannon diversity indicies for 11,353 genes both before and after infection with both (XLSX)Click here for additional data file.S6 TableEstimates of \u03a8 and \u0394\u03a8 values calculated by MISO (Supplementary Section1.3.5 in (XLSX)Click here for additional data file.S7 TableA summary of the proportion of annotated events that were detected and determined to be significant (according to the criteria defined in Supplementary Section 1.3.5 in (XLSX)Click here for additional data file.S8 TableListeria or Salmonella. All tests were done using an iterative gene ontology analysis as described in Supplementary Section 1.3.5 in Top gene ontology categories for genes that have significant RNA processing changes after infection with either (XLSX)Click here for additional data file.S9 TableA summary of the proportion of annotated evenst that were detected and determined to be significant (according to the criteria defined in Supplementary Section 1.3.5 in (XLSX)Click here for additional data file.S10 TableListeria or Salmonella, as described in Supplementary Sections 1.2.3 and 1.3.10 in 2 fold change estimates and p-values for differential expression of miRNAs after both 2 hours and 24 hours of infection, tested by DESeq2 as described in Supplementary Section 1.3.10 in Details of short RNA-seq data for 6 samples sequenced after both 2 hours and 24 hours of infection with either (XLS)Click here for additional data file."} +{"text": "Accumulating evidence suggests a cardioprotective role of pacing postconditioning (PPC) maneuvers in animal models and more recently in humans. The procedure however remains to be optimized and its interaction with physiological systems remains to be further explored. The renin angiotensin system (RAS) plays a dual role in ischemia/reperfusion (I/R) injury. The interaction between RAS and PPC induced cardiac protection is however not clearly understood. We have recently demonstrated that angiotensin (1\u20137) via Mas receptor played a significant role in PPC mediated cardiac protection against I/R injury.The objective of this study was to investigate the role of angiotensin converting enzyme (ACE)\u2014chymase\u2014angiotensin II (Ang II)\u2014angiotensin receptor 1 (AT1) axes of RAS in PPC mediated cardiac protection.Isolated rat hearts were subjected to I/R (control) or PPC in the presence or absence of Ang II, chymostatin , ACE blocker (captopril) or AT1 antagonist (irbesartan). Hemodynamics data was computed digitally and infarct size was determined histologically using TTC staining and biochemically by measuring creatine kinase (CK) and lactate dehydrogenase levels.Cardiac hemodynamics were significantly (P<0.001) improved and infarct size and cardiac enzymes were significantly (P<0.001) reduced in hearts subjected to PPC relative to hearts subjected to I/R injury. Exogenous administration of Ang II did not affect I/R injury or PPC mediated protection. Nonetheless inhibition of endogenously synthesized Ang II protected against I/R induced cardiac damage yet did not block or augment the protective effects of PPC. The administration of AT1 antagonist did not alleviate I/R induced damage. Interestingly it abrogated PPC induced cardiac protection in isolated rat hearts. Finally, PPC induced protection and blockade of locally produced Ang II involved enhanced activation of ERK1/2 and Akt components of the reperfusion injury salvage kinase (RISK) pathway.This study demonstrate a novel role of endogenously produced Ang II in mediating I/R injury and highlights the significance of AT1 signaling in PPC mediated cardiac protection in isolated rodents hearts ex vivo. The interaction between Ang II-AT1 and PPC appears to involve alterations in the activation state of ERK1/2 and Akt components of the RISK pathway. Cardiovascular diseases are major health concerns worldwide and coronary heart disease (CHD) continues to be a leading cause of death . IntermiThe renin-angiotensin system (RAS) is a master regulator of cardiovascular physiology and remolding . Key effAll chemicals used in this study were purchased from Sigma Aldrich unless otherwise indicated. The following drugs were used in the study and drug doses were selected based on previously published studies: Ang II (100nM) , AT1 bloThe study was approved by Health Science Center, Kuwait University Ethics Committee for animal use. The study was conducted according to the laboratory\u2019s animal care guidelines at Kuwait University, Kuwait in accordance with the international standards of animal care. Rats were maintained at 22\u00b0C on 12-hr light/dark cycle (7 am\u2013 7 pm) and water and food were available ad libitum. Rats were anesthetized with intraperitoneal injection of sodium pentobarbital (60 mg/kg) and heparin (1000 U/kg) was administered through the femoral vein, then were sacrificed by cervical dislocation.2 (5%) and O2 (95%). Pacing electrodes were placed on RA appendage to keep the heart beating at physiological rat heart rate. Regional ischemia was induced by occluding left anterior descending (LAD) coronary artery for 30 min. Preload was kept constant at 6 mmHg under basal controlled conditions and perfusion pressure (PP) at 50 mmHg throughout the experimental procedure in all protocols. The perfusion pressure was measured downstream from a branch of the aortic cannula using a \u201cstatham pressure transducer\u201d (P23 Db). Constant PP was ensured electronically by means of the perfusion assembly ), an effective system for the accurate adjustment of PP between 5 mmHg to 150 mmHg with \u00b1 1 mmHg accuracy level.Hearts isolated from male Wistar rats weighing between 270\u2013300 g were used for this study. Heart cannulation and perfusion were performed as described previously . O. O54]. OIn conclusion this study identifies a previously undiscovered role ACE and chymase induced synthesis of endogenous Ang II in mediating I/R cardiac injury and highlights the significance of ACE and chymase blockage as a potential therapeutic target in the treatment of myocardial infarction. It further characterizes a novel role of AT1 in PPC mediated cardiac protection of rodent hearts ex vivo. Finally it demonstrates that the RISK pathway is a downstream target of PPC and ACE-chymase-Ang II-AT1 signaling."} +{"text": "S2 Fig(A) Annotation Edit Distance (AED) distribution of gene models in the first annotation set after eliminating entries with AED = 1. (B). AED distribution of gene models in the second annotation after eliminating entries with AED = 1. (C) Proportion of gene models with protein domain hits in different heterokonts .(JPG)Click here for additional data file."} +{"text": "The dopamine hypothesis of schizophrenia has been extensively proposed as a neurobiological mechanism that explains the relationship between schizophrenic symptoms and hyperdopaminergic states. This hypothesis is supported by direct and indirect evidence, and it mainly postulates that antipsychotics act blocking dopamine receptors. When focusing on delusional disorder patients, especially delusional disorder somatic type, a great effort towards the search for a biological basis of treatment response has been recently demonstrated. Thus, the main goal of this systematic review was to examine the evidence explaining the biological underpinnings of treatment response in delusional disorder.A systematic review was performed using Pubmed, Scopus and PsycINFO databases (from 1990 to October 2017), according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. The following search terms were used: [(\u2018treat*\u2019 OR \u2018therap*\u2019 OR \u2018biol*\u2019) AND ]. This systematic computerized search was completed by additional studies hand-checked through reference lists from the included studies and review articles. Studies were only included if the met our inclusion criteria: (a) the International Classification of Diseases (ICD) or Diagnostic and Statistical Manual of Mental Disorders (DSM) diagnosis for delusional disorder, (b) be published in peer-reviewed journals, (c) in English, German or Spanish, (d) and reporting a hypothesis for the biological basis of treatment response in delusional disorder, irrespective of method and study design. Exclusion criteria were: (a) studies including organic delusional disorder or (b) somatic delusions secondary to other psychiatric diagnoses. The literature search strategy, data extraction and synthesis was conducted independently by two authors . When disagreement, it was solved by consensus.A total of 59 articles were identified, of which 12 met our inclusion criteria. Four hypotheses were addressed: (1) Dopaminergic dysfunction (n=4): ziprasidone-induced supersensitivity psychosis by chronic blockade of D2 Dopamine Receptor (DRD2) (n=1); pretreatment levels of plasma homovallinic acid (pHVA) (n=1); dopamine transporter (DAT) dysfunction (n=1) and effectiveness of aripiprazole (DRD2 agonist) (n=1). (2) Serotonergic dysfunction (n=6): drug occupancy in 5-HT1A and 5-HT2A receptors (n=3) and efficacy of 5-HT2 antagonists (n=3). Brain dysfunction (n=7): hypoperfusion in cerebral blood flow in temporal and parietal lobes, left side (n=5), right side (n=1) and lack of basal ganglia and subcortical gray matter lesions (n=1). Genetic evidence (n=1): implications of DRD2 Ser311Cys, DRD3 Ser9Gly and TH VNTR polymorphisms.The strongests biological contributors for treatment response in delusional disorder seem to be those implicating monoaminergic systems, particularly dopamine and serotonergic neurotransmitters. Although the low level of evidence, the serotonergic dysfunction may be associated with response rates, especially in delusional disorder somatic type. The link between genetic variants of dopamine receptors and neuroimaging findings in delusional disorder may open new avenues for the search of the biological underpinnings of treatment response. The evidence for an integrated model involving dopamine and serotonin systems bears further investigations."} +{"text": "In an effort to understand the regulatory role of Oct4A in tumor biology, we employed the use of an ovarian cancer shRNA Oct4A knockdown cell line (HEY Oct4A KD) and a global mass spectrometry (MS)-based proteomic analysis to investigate novel biological targets of Oct4A in HEY samples . Based on significant differential expression, pathway and protein network analyses, and comprehensive literature search we identified key proteins involved with biologically relevant functions of Oct4A in tumor biology. Across all preparations of HEY Oct4A KD samples significant alterations in protein networks associated with cytoskeleton, extracellular matrix (ECM), proliferation, adhesion, metabolism, epithelial-mesenchymal transition (EMT), cancer stem cells (CSCs) and drug resistance was observed. This comprehensive proteomics study for the first time presents the Oct4A associated proteome and expands our understanding on the biological role of this stem cell regulator in carcinomas.Oct4A is a master regulator of self-renewal and pluripotency in embryonic stem cells. It is a well-established marker for cancer stem cell (CSC) in malignancies. Recently, using a loss of function studies, we have demonstrated key roles for Oct4A in tumor cell survival, metastasis and chemoresistance in Ovarian cancer (OC) is the most lethal of all the gynaecological malignancies with a five-year mortality rate of >70%24The presence and importance of CSCs in different cancer scenarios including OC has been accumulating for the last ten years. However, the origin and the biological identity of CSCs associated proteome still remains unclear. Several potential indirect mechanisms of CSC regulation have been proposed; of particular interest are the Notch, Hedgehog, Janus activated kinase/Signal transduction and activator of transcription (JAK/STAT), anti-apoptotic and drug-resistant pathways568911Oct4 is a transcription factor which maintains self-renewal and pluripotency in embryonic stem cells and primordial germ cells131413151319MS-based proteomics has essentially changed the way by which malignant initiation and progression is investigated by its ability to identify and monitor thousands of proteins and post-translational modifications23Using a large-scale, label-free quantitative MS-based proteomic profiling approach, this study for the first time identified novel proteins and/or peptides which are associated specifically with Oct4A in the HEY ovarian cancer cell line and an associated mouse xenograft model. By identifying specific protein targets and select protein networks associated with differential expression of Oct4A, this study aimed to contribute to our knowledge of the biological traits driven specifically by Oct4A in OC and potentially other tumor models.2 and routinely checked for mycoplasma infection.The development of the HEY vector control and HEY Oct4A KD cell lines has been described previously2 as described previouslyOvarian cancer cell lines were treated with paclitaxel and cisplatin at GI50 concentrations (50% growth inhibitory concentrations) for 72\u2009hrs at 37\u2009\u00b0C in the presence of 5% COAnimal studies were carried out as described previouslyHEY vector control and HEY Oct4A KD cells were maintained in RPMI-1640 growth media as described previouslyFor tumor xenograft samples, tumors were produced by intraperitoneal (i.p) injection of HEY Oct4A KD and HEY vector control cells into Balbc/c nude mice as previously describedg for 5\u2009min, 2000\u2009\u00d7\u2009g for 10\u2009min) CM concentrated by centrifugal ultrafiltration as described previouslyConditioned media (CM) collected from sub-confluent (80%) HEY vector control and HEY Oct4A KD cells grown in RPMI-1640 (serum-free) were centrifuged and Ratio of spectral count (Rsc) were determined as previously described2930https://cran.r-project.org/web/packages/gplots/index.html) package in R software. Raw data set of proteins identified in vector control and HEY KD samples is described in Identification of entriched protein networks in cell lysates, secretomes and xenografts of Oct4AKD vs vector control was performed by STRING 10.0 software-\u0394\u0394Ct). The primer set of Oct4A, vimentin (VIM), plectin (PLEC), TUBB2A and the house keeping gene 18S are described in Quantitative real-time PCR was performed as described previouslyImmunohistochemistry analysis of mouse tumors was performed as described previously45We have recently shown knockdown of Oct4A in a HEY cell line by small hairpin (sh)RNA technology2727Following data collection and bioinformatics analyses, proteins which were not differentially expressed (p\u2009<\u20090.05) in HEY Oct4A KD samples when compared to HEY vector control samples were eliminated. Proteins identified as keratins were also removed from analysis based on known contaminants involved in proteomics analysisA correlation plot between the samples is presented in From the selection criteria stipulated in Compared to HEY vector control samples, a total of 16 differentially up regulated cellular proteins were identified in HEY Oct4A KD samples . ProteinAnalysis of the secretome revealed a total of 28 proteins were differentially suppressed in HEY Oct4A KD samples compared to HEY vector control samples . When clTwenty-eight secreted proteins were differentially over-expressed in HEY Oct4A KD conditioned media samples when compared to HEY vector control samples . SecreteA total of 72 proteins were differentially suppressed in xenograft tumors derived from HEY Oct4A KD cells when compared to xenograft tumour samples derived from HEY vector control cells . The incThirty-nine proteins were identified to be differentially elevated in tumor xenografts derived from HEY Oct4A KD cells compared to tumors derived from HEY vector control cells. Xenograft proteins which were identified to have functional roles in cellular growth, cellular metabolism, apoptosis, tumor suppression and oxidative stress response were the most frequently up regulated in HEY Oct4A KD tumor xenografts. This was followed by proteins involved with the cytoskeleton, calcium homeostasis and drug resistance. The most up regulated protein identified in HEY Oct4A KD xenograft tumor samples was the apoptosis-associated protein POTEF (Rsc 23.0). Other up regulated tumor xenograft proteins identified to be of interest included the tumor suppression-related protein Alpha amylase (AMY2A) (Rsc 7.3), the cellular growth-associated protein Eukaryotic translation initiation factor 4A2 (EIF42A) (Rsc 5.9), the drug resistance-associated protein Collagen alpha 3 (VI) chain (COLGA3) (Rsc 3.7) and the cellular metabolism-related protein Isocitrate dehydrogenase (IDH2) (Rsc 2.5). Up regulated xenograft tumor proteins and their proposed cancer-related classifications are listed in To identify protein networks and clusters associated with differentially expressed proteome profiles from HEY vector control and Oct4A KD cellular , secretome and xenograft , we performed protein-protein interaction analyses by STRING 10.0To validate the expression of selected proteins from proteomic profiling between the HEY Oct4A vector control and HEY Oct4A KD populations, RT-PCR and immunohistochemistry were carried out on a subset of proteins in Oct4A vector control and Oct4A KD cells and associated xenografts . The thrFor RT-PCR analysis, the expression of Oct4A was significantly greater in HEY vector control compared to HEY KD cells . This elWe have previously shown that the expression of Oct4/Oct4A is significantly enhanced in ovarian cancer cells in response to paclitaxel and cisplatin treatments34We have previously demonstrated that suppression of Oct4A in the HEY cell line is sufficient to impact on OC tumorigenesis, metastasis and chemoresistance. Characteristics which were notably affected included cellular proliferation, adhesion, migration, invasion, increased sensitivity to chemotherapy treatment and overall decreased tumor initiating ability and metastasis in mouse models2727Overall, the data indicated Oct4A to be a key regulator of cytoskeleton/ECM remodelling besides its tumorigenic role that we have described previously27More recently, TUBB2A has been described as having a role in regulating neuronal proliferation and migration38Among other cytoskeleton proteins, PLEC a member of plakin family was significantly down regulated in Oct4A knockdown tumor xenograft and secretomes4445The proteomic profiling of HEY Oct4A KD cells also demonstrated a loss of Mitogen activated protein kinase/Extracellular signal regulated kinase (MAPK/ERK) pathway in Oct4A knocked down tumor xenografts and secretomes. PLEC have been linked with MAPK/ERK pathway with respect to the migratory biology of keratinocytes and head and neck squamous cancer4650We have recently shown that suppression of Oct4A in HEY cell line resulted in the loss of \u03b1v and \u03b12 family of integrins53in vivo invasion and tumor development that we have reported previouslyProtein secretion by ovarian tumor cells has been shown to result in autocrine and paracrine signalling that defines cell growth, migration and the makeup of extracellular environmentin vitro and in vivo27Proteins found to be up regulated in HEY Oct4A KD were primarily associated with cellular survival. These categories included cellular proliferation, lipid metabolism, cellular metabolism, cellular growth and oxidative stress. This included cytoskeleton (e.g. TWF1), cellular growth (EIF42A), cellular metabolism (IDH2) and oxidative stress (HBA1). Interestingly, despite the up regulation of these proteins, HEY Oct4A KD cells derived tumors displayed an overall reduced growth and tumorigenic potential in mouse models2761http://icgc.org), EBI Expression Atlas (http://www.ebi.ac.uk) and The Cancer Genome Atlas which focused on ovarian tumors/tissues, suggest over expression of Oct4 in ovarian tumors. Furthermore, Oncomine indicates increased Oct4 gene expression (fold change 6.313) in ovarian tumors in comparison to normal ovarian tissues. Even though VIM/PLEC/TUBB2A demonstrated high expression in ovarian tumors (top 1%), no comparison with normal ovarian tissue exists in these databases so no enrichment/fold change can be indicated in tumors. These data support an association of Oct4A with cytoskeletal-ECM network, the key findings of this large-scale proteomics study. The attenuation of tumorigenic phenotype of HEY cells resulting from the knock down of Oct4A shown in our previous study further support these findings27This study has for the first time identified the global proteome profile associated with Oct4A in ovarian cancer, targeting cellular, secretome and tumor xenograft subsets. These findings along with the validation of key cytoskeletal and ECM proteins support the diminutive changes of proteins associated with cytoskeleton and ECM as major targets of Oct4A knockdown. In addition, gene/protein data using ICGC , Lactate dehydrogenase live type A (LDHA), Phospho fructo-kinase (PFK), Pyruvate dehydrogenase (PDH)) and (fatty acid synthesisFatty acid synthase (FASN)) to diminish in response to Oct4A knockdown. Overall, the protein changes observed are highly complex but the networks and results of this current proteomic analysis support the findings of our previous studies performed This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of the Laboratory Animals of the National Health and Medical Research Council of Australia. The experimental protocol was approved by the Ludwig Institute/Department of Surgery, Royal Melbourne Hospital and University of Melbourne\u2019s Animal Ethics Committee (Project-006/11), and was endorsed by the Research and Ethics Committee of Royal Women\u2019s Hospital Melbourne, Australia.How to cite this article: Samardzija, C. et al. Knockdown of stem cell regulator Oct4A in ovarian cancer reveals cellular reprogramming associated with key regulators of cytoskeleton-extracellular matrix remodelling. Sci. Rep.7, 46312; doi: 10.1038/srep46312 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations."} +{"text": "Glycogen debranching enzyme (AGL) and Glycogen phosphorylase (PYG) are responsible for glycogen breakdown. We have earlier shown that AGL is a regulator of bladder tumor growth. Here we investigate the role of AGL in non-small cell lung cancers (NSCLC). Short hairpin RNA (shRNA) driven knockdown of AGL resulted in increased anchorage independent and xenograft growth of NSCLC cells. We further establish that an increase in hyaluronic acid (HA) synthesis driven by Hyaluronic Acid Synthase 2 (HAS2) is critical for anchorage independent growth of NSCLC cells with AGL loss. Using gene knockdown approach against HAS2 and by using 4-methylumbelliferone (4MU), an inhibitor of HA synthesis, we show that HA synthesis is critical for growth of NSCLC cells that have lost AGL. We further show NSCLC cells without AGL expression are dependent on RHAMM for HA signaling and growth. Analysis of NSCLC patient datasets established that patients with low AGL/high HAS2 or low AGL/high RHAMM mRNA expression have poor overall survival compared to patients with high AGL/low HAS2 or high AGL/low RHAMM expression. We show for the first time that loss of AGL promotes anchorage independent growth of NSCLC cells. We further show that HAS2 driven HA synthesis and signaling via RHAMM is critical in regulating growth of these cancer cells with AGL loss. Further patients presenting with low AGL and HAS2 or RHAMM over expressing tumors might present the ideal cohort who would respond to inhibitors of HA synthesis and signaling. Amylo-alpha-1-6-glucosidase-4-alpha-glucanotransferase (AGL) along with glycogen phosphorylase (PYG) breaks down glycogen in human cells . GSDIII By thorough experimentation we validated that AGL's enzymatic function does not play a role in regulating tumor growth . We alsoHere we investigate the role of AGL in non-small cell lung cancer. We show that loss of AGL promotes aggressive anchorage independent and xenograft growth of NSCLC cells. This is the first report showing AGL as a novel regulator of NSCLC anchorage independent growth. We also illustrate that the AGL low NSCLC cells are vulnerable to inhibition of HAS2 dependent HA synthesis and HA signaling via RHAMM.nd shRNA construct against AGL (shAGL') which targeted the 3'UTR region (TRCN0000419324). Loss of AGL using this construct also resulted in increased anchorage independent growth of H2122 and H358 NSCLC cell lines proving that the effect we see on anchorage independent growth is specific to AGL loss for our study. We carried out AGL knockdown (shAGL) in these cells using the previously described and validated AGL shRNA construct TRCN0000035082 from Sigma-Aldrich [AGL loss . HoweverAGL loss . Next weAGL loss , howeverAGL loss .AGL has two known enzymatic functions, glucosidase and transferase , 7. We gNext we knocked down glycogen phosphorylase isoforms (brain and liver) in H2122 and A549 cells Figure . GlycogeHAS2 driven HA synthesis is known to promote anchorage independent growth , 10. We We carried out transient depletion of HAS2 in NSCLC H2122 and A549 cells with and without AGL expression using a siRNA construct which we have previously validated [Next we tested cell proliferation of A549 and H2122 +/\u2212 AGL cells after transient transfection with control siRNA and siRNA against HAS2 over 5 days. As we have previously seen , loss ofAfter establishing that the AGL knockdown cells synthesize more HA, we subjected the AGL knockdown H2122 and A549 cells to treatment with 4MU, an inhibitor of HA synthesis . TreatmeTo address the role of the two major HA receptors CD44 and RHAMM , 13 in tWe carried out RHAMM knockdown with a previously validated siRNA in A549 The role of AGL as a predictor of NSCLC patient outcome is not known. We analyzed 4 independent patient cohorts of NSCLC adenocarcinoma patients with Stage I and II tumors (N = 555) \u201318 to deWe have shown that HAS2 expression is elevated with loss of AGL. Next we investigated whether AGL and HAS2 in combination can predict patient outcome. The primary objective here was to determine if such expression levels could eventually be used to identify the optimal patient cohort who may be enrolled in future clinical trials with inhibitors of HA signaling. The secondary objective was to lend credence to the hypothesis that AGL affects tumor biology by HAS2 mediated HA synthesis. We segregated patients with high AGL and low HAS2 (AGL+/HAS2-) mRNA expression from patient with low AGL and high HAS2 mRNA expression (AGL-/HAS2+). Kaplan\u2013Meier survival showed that patients with AGL-/HAS2+ have poor overall survival compared to AGL+/HAS2- patients Figure with staSimilarly we looked at whether AGL and RHAMM, the HA receptor important for growth of AGL knockdown NSCLC cells, stratify NSCLC patient outcome. Kaplan\u2013Meier survival show that patients with low AGL and high RHAMM mRNA expression (AGL-/RHAMM+) have poor overall survival compared to patients with high AGL and low RHAMM expression (AGL+/RHAMM-) with significant P values in 3 of the 4 patient cohorts analyzed Figure . FurtherHere for the first time we show that glycogen debranching enzyme (AGL) regulates NSCLC tumor growth. We further validate that AGL regulates NSCLC growth independent of its enzymatic function which is consistent with our previous findings in the bladder tumor model . Here weWe have earlier shown that in bladder tumors, loss of AGL promotes rapid anchorage dependent and independent growth of cancer cells . HoweverWe show that with loss of AGL there is an increase in HAS2 expression and hyaluronic acid synthesis. It is well established that HAS2 driven HA synthesis or HA in general plays a major part in promoting anchorage dependent and independent growth , 10. InhAnother important aspect which merits discussion is HA size and their impact on tumor growth. Hyaluronic Acid Synthases are known to make HA of different sizes . HAS2 whIt is still unclear how loss of AGL results in increased HAS2 expression and HA synthesis. Our previous study in the bladder tumor model indicated metabolic reprogramming and increased glucose uptake and glycolysis with AGL loss may play an important role in increasing HA synthesis , 4. GlucWe have shown that AGL, in combination with HAS2 or RHAMM, predict NSCLC patient outcome. Patients with low AGL and high HAS2 or RHAMM expression had poor overall survival compared to patients with high AGL and low HAS2 or RHAMM expression. This provides credence to our findings that HA synthesis and signaling is important for tumors with low AGL expression. This data will also help in the selection of the ideal patient cohort for future intervention with inhibitors of HA synthesis and signaling.4MU is a HA synthesis inhibitor which has been well studied in the field of cancer biology . A greatin vitro and in vivo setting. Short hairpin RNA (shRNA) sequence 5'-CCGGCCCTTGCCAATCAGTTAGAATCTCGAGATTCTAACTGATTGGCAAGGGTTTTTG-3' [NSCLC cell lines H358, H2122 and A549 were obtained from the American Type Culture Collection and cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS) as recommended by ATCC. These three NSCLC cell lines were chosen for the study because they show an induction in growth with AGL loss; therefore serve as good model cell lines to study AGL biology in NSCLC. These cell lines were also chosen because they are known to form xenografts and hence will allow us to study AGL biology in Aldrich) , 26 hencHAS1-3 mRNA expression was determined by the \u0394\u0394CT method with GAPAntibodies used for westerns were anti-AGL , Actin , CD44 , RHAMM . HRP labeled mouse or rabbit secondary antibodies were used for chemiluminescence detection with ECL reagents as previously described \u20135.Anchorage dependent and independent proliferation was measured as previously described , 4, 34. 3 cells per well in 96-well plates in triplicate for proliferation studies. CyQUANT\u00ae Cell Proliferation Assay (Invitrogen) was carried out according to manufacturer instructions to measure cell proliferation. To determine the effects of 4MU on cell viability, cells were plated as described, and treated with 4MU (600 \u03bcM) or 4MU (600\u03bcM) and HA(20\u03bcg/ml) together for 5 days. Cell viability was determined by CyQUANT Assay (Invitrogen).For anchorage dependent growth assay, cells with or without AGL expression were transfected with control siRNA or siRNA targeting HAS2 or RHAMM . 72hrs aFresh media is applied 48hrs after HAS2 siRNA transfection in AGL knockdown cells followed by HA analysis by ELISA 24 hrs later. Cells with and without AGL are grown to 60-65% confluency followed by fresh media addition with increasing concentrations of 4MU to evaluate the impact of 4MU on HA synthesis and secretion after 24hrs. HA ELISA was conducted as per manufacturer instructions using TECO\u00ae HA ELISA kit.6), H2122 (1.0\u00d7105) or A549 (2\u00d7106) cells stably expressing AGL shRNA or nontarget shRNA control (7 mice per group) in the right and left flanks of each mouse for subcutaneous tumor growth. Tumors were measured and tumor volumes calculated as described previously [2)/2. Animals without tumor take (i.e. measurement of 0) were excluded from tumor volume calculations.All animals used in this study were treated according to Institutional Animal Care and Use Committee (IACUC) guidelines. Four-week-old NCr nu/nu female mice from Charles Rivers or Envigo were injected with H358 in R (https://cran.r-project.org/).CEL files for the CAN/DF cohort were downloaded and proc"} +{"text": "Clinically useful molecular tools to triage gastric cancer patients are not currently available. We aimed to develop a molecular tool to predict gastric cancer risk in endoscopy-driven biopsies obtained from high-risk gastric cancer clinics in low resource settings.IRF4, ELMO1, CLIP4 and MSC.We discovered and validated a DNA methylation biomarker panel in endoscopic samples obtained from 362 patients seen between 2004 and 2009 in three high-risk gastric cancer clinics in Lima, Per\u00fa, and validated it in 306 samples from the Cancer Genome Atlas project (\u201cTCGA\u201d). Global, epigenome wide and gene-specific DNA methylation analyses were used in a Phase I Biomarker Development Trial to identify a continuous biomarker panel that combines a Global DNA Methylation Index (GDMI) and promoter DNA methylation levels of p < 0.0001). Promoter methylation of IRF4 (p < 0.0001), ELMO1 (p < 0.0001), CLIP4 (p < 0.0001), and MSC (p < 0.0001), is also associated with increasing severity from gastritis with no metaplasia to gastritis with metaplasia and gastric cancer.We observed an inverse association between the GDMI and histological progression to gastric cancer, when comparing gastritis patients without metaplasia , gastritis patients with metaplasia , and gastric cancer cases , respectively (IRF4, ELMO1, CLIP4 and MSC promoter methylation coupled with a GDMI>4 are useful molecular tools for gastric cancer risk stratification in endoscopic biopsies.Our findings suggest that Helicobacter pylori leading to chronic inflammation is a major attributable risk factor \u00d7 100.Global DNA methylation levels were determined by ELISA using the MDQ1, Imprintn = 10), gastritis samples misdiagnosed as cancer by endoscopists (n = 10) and cancer samples (n = 10). To validate these results we performed an unbiased epigenome-wide DNA methylation analysis to identify DMRs in gastric cancer samples (n = 295) from the Cancer Genome Atlas project (TCGA) and gastritis controls (n = 20) from Per\u00fa.The HumanMethylation450K DNA BeadChip assay was used to perform unbiased epigenome-wide DNA methylation analysis. Bisulfite modification of genomic DNA (2 \u03bcg) was performed with EpiTect Bisulfite Kit (QIAGEN) according to the manufacturer's protocol. We hybridized bisulfite converted DNA to the HumanMethylation450K array to identify differentially methylated regions (DMRs) in gastritis samples . We then intersected the statistically significant DMRs (FWER p < 0.05) that discriminate gastric cancer from gastritis in samples from Peru and TCGA.We imported the data into R using the illuminaio package . For datp < 0.05) common to both the DMR results from the Peru and TCGA epigenome-wide analyses: IRF4 (interferon regulatory factor 4); ELMO1 (encodes Engulfment and cell motility protein 1); CLIP4 (CAP-Gly domain containing linker protein family member 4); and MSC (encoding Musculin protein). We designed primers and probes to quantify promoter methylation of these four genes using fluorogenic quantitative methylation specific PCR (qMSP), as previously described [We selected four of the genes that had the greater variance and the largest number of CpGs in the DMR window, from the list of significant DMRs and gene-specific DNA methylation (n = 116) analyses and a Validation cohort for combined global and gene-specific DNA methylation analyses for whom we also had a diagnosis provided by the endoscopists. In addition, epigenome-wide analyses were performed in 295 gastric cancer samples and 11 normal gastric epithelium samples from the Cancer Genome Atlas (TCGA) project.The primary outcome indicator was a binary variable to identify gastric adenocarcinoma (GC) from gastritis cases. We also used the level of metaplasia as a proxy to identify gastritis patients with higher risk of developing GC. We conducted ordinary and logistic regression and analysis of variance, Fisher exact test and other tests of hypothesis to analyze the data. All data was analyzed and managed using STATA 13 and results with a"} +{"text": "Background: Right temporal lobe lesion syndrome elicitation presents a clinical challenge. Aside from occasional covert quadrantanopias, heralding elementary neurological deficits are absent.Aim: Isolated right and left temporal lobe stroke patients were analyzed for the panoply of known temporal and frontal cognitive and neuropsychiatric syndromes. Methods: Temporal lobe stroke patients were analyzed, derived from a dedicated cognitive stroke registry. Patients were screened by a validated bedside cognitive battery and a neuropsychological test battery, including the Bear Fedio Inventory for diagnosis of the Geschwind Gastaut (GG) syndrome, frontal network syndrome testing (FNS), emotional intelligence testing and delusional misidentification syndromes (DMIS). NIH stroke scores were documented and lesion location identified with the 3 dimensional digitized Cerefy coxial brain atlas. Exclusions were coma, encephalopathy and medication related effects.Results: Of 2389 patients analyzed, in patients with isolated right temporal lobe (IRT) stroke , the GG syndrome and FNS were present in all five. Other relatively frequent syndromes included DMIS in 4, visuospatial dysfunction in 2 and amusia in 2. No patient had a NIHSS greater than 1. The only elementary neurological sign was quadrantanopia in 3 patients. Lesion location was mid and lateral temporal lobe (n = 2), middle and mesial temporal lobe (n = 1) middle temporal lobe (n = 1) and lateral temporal lobe (n = 1). Comparison with isolated left temporal lobe (ILT) stroke revealed syndromes of aphasia (n = 4), alexia (n = 2), acalculia (n = 2), agnosia (n = 2), verbal amnesia (n = 1), none of which occurred in the IRT patients. The mean NIHSS scores of IRT (0.6) and ILT strokes (4.2) was different . The 2 \u00d7 8 Fisher Exact Test revealed significant differences for the clusters of syndromes occurring in the right and left isolated temporal lobe lesions (p = 0.00002).Conclusion: The GG syndrome, FNS and DMIS are prominent syndrome constellations in stroke patients involving the right temporal lobe and constitute the neurological deficit without heralding long tract signs. By extrapolation these syndromes may also be present in the general right hemisphere lesion population."} +{"text": "Oesophageal adenocarcinoma (OAC) is one of the ten most prevalent forms of cancer and is showing a rapid increase in incidence and yet exhibits poor survival rates. Compared to many other common cancers, the molecular changes that occur in this disease are relatively poorly understood. However, genes encoding chromatin remodeling enzymes are frequently mutated in OAC. This is consistent with the emerging concept that cancer cells exhibit reprogramming of their chromatin environment which leads to subsequent changes in their transcriptional profile. Here, we have used ATAC-seq to interrogate the chromatin changes that occur in OAC using both cell lines and patient-derived material. We demonstrate that there are substantial changes in the regulatory chromatin environment in the cancer cells and using this data we have uncovered an important role for ETS and AP1 transcription factors in driving the changes in gene expression found in OAC cells. Oesophageal adenocarcinoma is one of the ten most prevalent forms of cancer and is showing a rapid increase in incidence and yet exhibits poor survival rates. Understanding the molecular causes of this type of cancer will enable us develop more effective treatment strategies which will improve survival rates. Here we have investigated how the genes in cancer cells are packaged into chromatin. We then compare this packaging to normal cells and use this information to identify the molecular causes leading to changes in chromatin packaging in cancer cells. We have identified a regulatory factor called AP1 that acts as a molecular switch to alter gene expression and hence cause cells to adopt a cancer fate. Importantly either this regulatory factor or a coregulatory protein from the ETS family is upregulated in the majority of cancer cells. Our study has therefore uncovered an important regulatory pathway that is commonly activated in oesophageal adenocarcinoma cells. The incidence of oesophageal adenocarcinoma (OAC) in the Western world is increasing and five and ten year survival rates remain low ,2. In coSeveral studies have implicated different transcription factors as important drivers of oesophageal cancer, chiefly due to their overexpression in OAC cell lines and/or patient derived OAC samples. Well studied examples include GATA6 ,11, and In this study, we investigated the changes that occur in the regulatory chromatin landscape in oesophageal adenocarcinoma by an unbiased approach using ATAC-seq. We identified AP1 and ETS transcription factors as important regulators in OAC cells and targeted ChIP-seq analysis combined with knockdown experiments reinforced the role of the ETS protein ETV1 in driving OAC-specific gene expression programmes. Similarly, loss of function approaches validated a regulatory role for AP1. Our results therefore demonstrate an important role for AP1 in OAC and part of its action is through a regulatory module containing AP1 and PEA3 subfamily ETS transcription factors. Importantly one or both of these factors are commonly upregulated in patient-derived OAC samples, and both factors are implicated in regulating the active open chromatin environment in these cells.Our previous studies focussed on ETV4 and its role in OAC but also demonstrated that the closely related transcription factor ETV1 is upregulated in OAC . We wereDUSP6 promoter was demonstrated to be target for PEA3 family proteins in two previous ChIP-seq studies in ECC-1 endometrial carcinoma cells [DUSP6 promoter as a positive control to optimise the ETV1 ChIP protocol with only 8\u20139% found in promoter regions . ExampleG/CTCA which forms non-DNA binding heterodimers with endogenous JUN family members . We obse-50). The active chromatin state is usually coincident with the appearance of histone H3K27 acetylation. We therefore tested whether inhibition of AP1 activity by DN-FOS also affected the levels of this chromatin mark. At all four loci tested, DN-FOS reduced the levels of gene expression but with the exception of MMP1, failed to change the levels of H3K27ac , with 32% (233/738) also showing downregulation after AP1 inhibition , and >1.3 fold reduction in expression following DN-FOS expression which gave us a total of 58 genes which are high confidence direct AP1 targets. We then examined the expression of these genes in a panel of OAC samples and plotCollectively, this data identifies the AP1 transcription factor as an important player in driving the gene expression profiles found in OAC cells.DUSP6 promoter . Target gene expression was then examined and samples clustered according to similarity in expression patterns . This idTogether these results demonstrate that ETV1 binding occurs concomitantly with AP1 and is generally associated with chromatin which becomes more open and active in cancer cells. However, while some regions are specific to OE33 cells, others are commonly activated in several OAC-derived cell lines.FOSL2 (ETV1). Together these clusters represented the majority of the samples but 29% formed a third cluster which is characterised by high level expression of individual ETS and AP1 family subunits rather than coordinated upregulation of several family members. Overall patients with OAC therefore exhibited either coordinated upregulation of AP1 subunits or PEA3 subunits or combinations of individual subunits from the two transcription factor families.Our data from ChIP-seq and ATAC-seq studies in OAC-derived cell lines implicate ETV1 and AP1 transcription factors as important players in driving OAC-specific gene expression programmes. One prediction of these findings is that we should see greater expression of these transcription factors in patient-derived OAC samples compared to normal oesophageal tissue. AP1 is a heterodimeric transcription factor consisting of either homodimers of JUN family members or heterodimers of JUN family members with a FOS family member [reviewed in FOSL2 . The expFOSL2 . The firFOSL2 which is higher and searched for evidence for regulation by these transcription factors. First we merged the reads from all the tissue samples both normal and cancer. We then identified the top 50,000 peaks from these merged reads and performed principal component analysis (PCA). The normal samples (blue) cluster together whereas the tumour samples (red) separated from the normal samples . FurtherIHH locus S1 Fig) aEthical approval for collection of oesophageal tissue samples from patients at the Royal Albert Edward Infirmary, Wigan and the Salford Royal Hospital were granted by the ethics committees at Wrightington, Wigan and Leigh NHS Foundation Trust (2007) and Salford Royal NHS Foundation Trust (2010) respectively (04/Q1410/57). Patient consent was obtained in written form and signed by the patient and doctor.2) in epithelial cell media supplemented with 1% epithelial growth supplement. Biopsy tissue samples (~4 mm) were processed as described previously .The OE33 and OE19 cells were cultured in RPMI media supplemented with 10% foetal bovine serum. The FLO1, HET1A and 293T cells were cultured in DMEM culture media supplemented with 10% FBS. HEEPIC cells were grown on poly-l-lysine coated plates , or a non-targeting pool (Dharmacon) were used in 24 hr transfections prior to RNA extraction as described previously [SDHA, ALAS1, GAPDH, and HMBS using the delta CT method.Total cellular RNA was isolated from cell line and clinical tissue samples as described previously . When reeviously . For theeviously with theth or the 75th percentile (e.g 75th percentile + (1.5 x IQR) and 25th percentile\u2013(1.5x IQR)). Statistical significance was assessed using a 2 tailed t-test calculated using StatPlus Microsoft Excel unless otherwise specified.In knockdown experiments statistical significance was calculated using an unpaired two-tailed Student\u2019s T test with a two sample equal variance. Gene expression data comparing expression in different groups of samples are represented with boxplots generated using Gene-e software. Outliers are not shown but represent values >1.5 interquartile ranges from the 25For expression microarray analysis of HET1A, HEEPIC, and OE33 cells (plus/minus siETV1/ETV4), biological triplicate RNA samples for each condition were processed. The raw intensity files (CEL) were generated by processing 500 ng of total RNA on Affymetrix HTA 2.0 arrays, according to the manufacturer's instructions . The arrays were scanned through GENECHIP Scanner-7G . The CEL files generated by these arrays were converted into rma-gene-ful.chp and rma-alt-splice-dabg.chp files through Affymetrix Expression Console Software (version 1.3). The CHP files were analyzed through the Transcriptome Analysis Console v3.0 (TAC). Within the TAC software an excel spreadsheet for all conditions with the mean relative expression (n = 3) of all genes was generated. From this TAC also calculated fold change with between conditions. For all downstream analysis involving fold change genes this spreadsheet was used to generate gene lists for further analyses. Data are deposited in ArrayExpress (Accession number: E-MTAB-5163).To generate samples for RNA-seq analysis, we transduced OE33 cells with lentiviruses expressing a dominant-negative (DN) FOS (DN-FOS) construct (pInducer-DN-FOS) or lacking an insert (control)(pInducer) and grew for a further 12 hours. pInducer-DN-FOS was constructed by inserting a BamHI/EcoRI fragment from pBABE-puro-a-FOS/pAS2804 into the7 cells, 3 \u03bcg antibody ) and 30 \u03bcl Dynabeads were used per experiment. Parallel control experiments were run with ChIP-qPCR using rabbit IgG; Millipore (12\u2013370). Library preparation was performed using the TruSeq ChIP Sample Preparation Protocol (Illumina) and DNA libraries were sequenced using the HiSeq 2500 (Illumina).ChIP-qPCR and ChIP-seq were carried out as described previously . For ChISequencing tags/reads from the ETV1 ChIP-seq experiment in OE33 cells were aligned to the NBCI Build hg19 of the human genome with Bowtie v2.2.3 . Up to tATAC-seq data generation and analysis on OE19 and HEEPIC cell lines was performed as described previously for the HET1A, OE33 and FLO1 cell lines . Hierachical clustering was carried out using this software, and all clustering was using one minus Pearson\u2019s correlation unless otherwise specified. De novo motif discovery in ATAC-seq and ChIP-seq was carried out using HOMER [To visualise ATAC-seq data, normalised cleavage events across the differentially accessible regions were counted using HOMER to produng HOMER with thehttp://bejerano.stanford.edu/great/public/html/) [10 of the binomial p-value generated using GREAT software [Gene annotation was performed using HOMER to identc/html/) using NBsoftware .Tag density heatmaps and profiles were generated using HOMER using deS1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file.S3 Fig(PDF)Click here for additional data file.S4 Fig(PDF)Click here for additional data file.S5 Fig(PDF)Click here for additional data file.S6 Fig(PDF)Click here for additional data file.S7 Fig(PDF)Click here for additional data file.S8 Fig(PDF)Click here for additional data file.S9 Fig(PDF)Click here for additional data file.S10 Fig(PDF)Click here for additional data file.S11 Fig(PDF)Click here for additional data file.S12 Fig(PDF)Click here for additional data file.S13 Fig(PDF)Click here for additional data file.S14 Fig(PDF)Click here for additional data file.S15 Fig(PDF)Click here for additional data file.S1 TablePeaks are assigned to the gene with the nearest TSS.(XLSX)Click here for additional data file.S2 TableRelative expression in each cell line/condition, and fold changes and associated P-values for the indicated comparisons are shown in tab 1. Tab 2 shows the genes that are significantly changed upon siETV1 treatment and the changes in their expression following treatment of OE33 cells with DN-FOS constructs.(XLSX)Click here for additional data file.S3 TableColumns J-Q show normalized Tn5 cutting frequency within each indicated 500 bp window in each of the indicated cell lines. Peaks are assigned to the gene with the nearest TSS.(XLSX)Click here for additional data file.S4 TableData are shown as averages of three experimental replicates.(XLSX)Click here for additional data file.S5 TableColumns F-M show normalised Tn5 cutting frequency within each indicated ETV1 binding region in each of the indicated cell lines. The final column shows the clustering from (XLSX)Click here for additional data file.S6 TableInformation about samples used in ATAC-seq and for gene expression on the Fluidigm Biomark system are shown on separate tabs.(XLSX)Click here for additional data file.S7 TableColumns H-P show normalised Tn5 cutting frequency within each indicated chromatin regions in each of the indicated tissue samples from normal tissue (N) or OAC tumour samples (T). Regions are included that show significant differential accessibility between the normal and the cancer samples.(XLSX)Click here for additional data file.S8 Table(XLSX)Click here for additional data file."} +{"text": "Human and murine MM cell lines cultured in osteogenic medium expressed alkaline phosphatase and formed mineralized bone-like nodules. Several human and mouse MM cell lines also expressed a number of osteoblast phenotype markers, including runt-related transcription factor 2 (RUNX2), osteopontin, osteonectin and bone sialoprotein mRNA and protein. Histological analysis of murine MM tumors identified areas of ossification within the tumor, similar to those observed in human MM biopsies. These data demonstrate the ability of MM to differentiate into another mesenchymal cell type and suggest that MM cells may contribute to the formation of the heterologous elements observed in MM tumors.Malignant mesothelioma (MM) is an aggressive serosal tumor associated with asbestos exposure. We previously demonstrated that mesothelial cells differentiate into cells of different mesenchymal lineages and hypothesize that osseous tissue observed in a subset of MM patients is due to local differentiation of MM cells. In this study, the capacity of human and mouse MM cells to differentiate into osteoblast-like cells was determined Malignant mesothelioma (MM) is a rare but aggressive primary tumor of the serosa associated with past exposure to asbestos34th century, Durante and Conheim presented the embryonal rest theory of cancer, which stated that remnants of embryonic tissue remain in adult organs and that a change in the surrounding environment would allow the embryonic tissue to proliferate and produce masses of cells that resemble fetal tissues910111213in vitro under the influence of osteogenic medium (OM)4It has been proposed that the mesenchymal components (osseous and cartilaginous) and the variability of the histological subtypes of MM are due to the capacity of mesothelial-derived cells to differentiate into multiple cell lineages of the embryonic mesoderm (termed multipotent)Analysis of a MM biopsy from a 27 year-old woman with childhood exposure to asbestos showed areas of mineralized bone formation within the tumor which waSimilar areas of bone tissue were identified in C57BL/6 MM tumors . Murine Three human and three mouse MM cell lines were examined. NO36, JU77 and AE5 cells displayed an epithelioid morphology whereas LO68, AB1 and AE17 cells exhibited a more spindle morphology . Followiin vitro (black staining).Cell lines expressing high AP levels also stained more intensely with von Kossa, demonstrating the presence of mineralized nodules . AP leveThe pathway leading to bone formation has been well describedAll MM cell lines analyzed expressed RUNX2, SPARC and SPP1 mRNA and protein at day 0 and throughout the nodule formation assay . The proin vitro, LO68 cells, which do not express high levels of alkaline phosphatase or mineralize when cultured in OM, were grown to confluence and incubated in NO36 conditioned medium (NO36 CM), OM, OM\u2009+\u2009NO36 CM or 10%FCS medium for 21 days. NO36 CM stimulated similar levels of AP production in LO68 cells compared with OM and OM + NO36 CM or AB1 (AP intermediate expression) syngeneic murine MM cells subcutaneously into the hind flank of C57BL/6 or BALB/c mice respectively. AB1 tumors followed a similar pattern of growth in both treatment and control groups , or what has more recently been described as mesothelial to mesenchymal transition (MMT), and differentiate into cells of different phenotypes42829303233343536438In the current study, we used MM cell lines of different histological subtypes to examine their ability to differentiate into osteoblasts under appropriate culture conditions. We had hypothesized that the mesenchymal or sarcomatoid phenotype would have the greatest capacity for differentiation as cells generally differentiate into a mesenchymal phenotype before differentiating into osteoid cells. Interestingly, although each MM cell line demonstrated a capacity for osteoblast-like differentiation, epithelioid MM cells had the greatest capacity to differentiate. Whether or not the better prognosis associated with the epithelioid MM phenotypeWe clearly show mRNA and protein expression of several key osteoblast markers. Interestingly, bone-specific RUNX2 was expressed in unstimulated (day 0) MM cells, consistent with our observation in normal mesothelial cellsIt is unclear what drives the osteoblast differentiation within the tumor. It has been suggested that a soluble mediator, perhaps induced or produced by MM cells, stimulates the formation of bonein vitro compared with the other two cell lines examined and AB1 cells demonstrated a lower differentiation capacity in vitro. Therefore the effect of DEX treatment on the growth and osteoid formation of these cells were compared in vivo. DEX treatment did not induce bone formation in either cell line, however tumor growth was significantly increased in DEX treated AE5 cells with evidence of dermal invasion and abundant collagen. DEX has been shown to stimulate connective tissue growth factor expression in mouse fibroblasts which leads to increased collagen productionDexamethasone (DEX) is known to induce the osteogenic differentiation of pre-osteoblasts, mesenchymal stem cells and embryonic stem cells at physiological levels4546in situ may be a treatment option. Differentiation therapy has already been used in several different cancers to induce terminal differentiation in cancer stem cells to decrease cell proliferation and halt tumor progressionExactly how the unusual histological variants (heterologous elements) of MM may affect patient prognosis is currently unknown. Demirag and colleaguesThis study provides further evidence for the multipotent nature of mesothelial-derived cells4Three previously established human MM cell lines isolated from pleural effusions of MM patients51Human and murine MM cells were maintained in either standard medium (DMEM with high glucose 4.5\u2009mg/L) supplemented with 10% foetal calf serum (FCS), 4\u2009mM L-glutamine, 100,000 units/L penicillin and 50\u2009mg/mL streptomycin) or osteogenic medium as per the manufacturer\u2019s instructions and DNase treated prior to cDNA synthesis (Invitrogen Life Technologies). cDNA was synthesized using a Superscript III first strand synthesis kit (Invitrogen Life Technologies). The mRNA expression of human osteoblast phenotype markers was determined using gene specific primers and standard PCR conditions as previously reported\u00ae Novex\u00ae bis-tris gel (Invitrogen Life Technologies) and protein immobilized on a PVDF membrane . Membranes were blocked overnight at 4\u2009\u00b0C in 5% skim milk/TBS-T (TBS with 0.05% Tween 20). Following serial washing the membranes were incubated with either mouse RUNX2 , rat secreted protein, acidic, cysteine rich , rat secreted phosphoprotein 1 or anti-\u03b1-tubulin . Membranes were washed and incubated with HRP-conjugated anti-rabbit or anti-mouse antibody . Membranes were visualized using the chemiluminescent peroxidase substrate and hyperfilm ECL .Cells were lysed in breaking buffer as previously describedAll animal experiments were undertaken with the approval of the Animal Ethics Committee of the University of Western Australia (RA/05/100/505) in accordance with National Health and Medical Research Committee guidelines and regulations.6 AB1 and 3.5\u2009\u00d7\u2009106 AE5 syngeneic cells were injected subcutaneously into the right flank of either BALB/C (AB1) or C57BL/6 (AE5) mice respectively and tumor growth monitored by taking perpendicular measurements using microcalipers. Once tumors reached approximately 1\u2009mm2, 10 animals per group were treated with either dexamethasone or saline via intra-peritoneal injection for 22 days. Once tumors had either reached 100\u2009mm2 or had grown for a pre-determined amount of time they were removed and paraffin embedded for histological evaluation.Approximately, 2\u2009\u00d7\u200910All human samples were obtained and used in accordance with National Health and Medical Research Committee guidelines and regulations and ethics approved by the Sir Charles Gairdner Ethics Committee.Formalin-fixed, paraffin-embedded (FFPE) lung tissue sections from a 27 year old female patient with a confirmed diagnosis of malignant mesothelioma were obtained from retrospective diagnostic biopsy specimens from PathWest Laboratory Medicine . Analysis of murine tumors was performed on archival tumor tissue samples obtained from 39 MexTAg mice and 11 C57BL/6 wild-type (WT) mice inoculated with asbestos as describedAll data are presented as mean\u2009\u00b1\u2009standard error of mean (SEM). Comparisons between individual time points were performed using a one sample t-test or a one way ANOVA using a Tukey\u2019s Multiple Comparison test as appropriate to determine significance. A p value of less than 0.05 was considered significant.How to cite this article: Lansley, S. M. et al. A Subset of Malignant Mesothelioma Tumors Retain Osteogenic Potential. Sci. Rep.6, 36349; doi: 10.1038/srep36349 (2016).Publisher\u2019s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations."} +{"text": "Avibacterium paragallinarum, Ornithobacterium rhinotracheale (ORT), Mycoplasma synoviae (MS), Mycoplasma gallisepticum (MG) and Avian pathogenic Escherichia coli (APEC) are involved in the respiratory disease complex in birds In recent past, the respiratory infection has emerged as a great challenge to the poultry farmers. Various pathogens including Avian pneumovirus (APV), Avian influenza virus (AIV), Infectious bronchitis virus (IBV) and Newcastle disease virus (NDV), Specifications TableValue of the data\u2022These data represents the first WGS metagenomic sequences of genomic DNA isolated from respiratory tract of broiler.\u2022Metagenome data are used to identify different pathogens responsible for causing respiratory disease complex.\u2022These metagenome are valuable for the study and comparison of microbial communities among different types of birds.\u2022The additional metagenome data will provide information about virulence genes of respiratory pathogens for prevention and control of disease.1Mycoplasma synoviae, Mycoplasma gallisepticum, Avibacterium paragallinarum, Ornithobacterium rhinotracheale Escherichia coli were most important pathogens for respiratory disease complex.Metagenomics is newer technique to overcome the limitations of culture dependent microorganism studies 22.1Approximately 500\u202f\u00b5l to 1.2\u202fml of respiratory lavage was collected from each bird and transferred to a sterile 1.5-ml microcentrifuge tube and stored at -20\u00b0C until further processed for genomic DNA extraction.2.2Genomic DNA (gDNA) was extracted by using a commercially available DNA isolation kit according to the manufacturer's instructions. Genomic DNA concentration was measured using Qubit\u00ae dsDNA HS Assay Kit (Life Technologies). Total 100\u202fng of DNA was used to prepare the library using the Ion Xpress\u2122 Plus Fragment Library Kit (Cat. No. 4471269) and Ion Xpress\u2122 Barcode Adapters 1\u201316 (Cat. No. 4471250). The final library was used for emulsion PCR, enrichment and sequencing. The Ion PGM\u2122 Hi-Q\u2122 Sequencing Kit and 318 chip were used for sequencing reactions, following the recommended protocol.2.3https://www.ncbi.nlm.nih.gov/sra/?term=PRJNA339659).The metagenome sequences of broiler have been deposited in the National Center for Biotechnology Information (NCBI) BioSample Submission Portal as Bioproject PRJNA339659 and SRA accession number SRR5997823, SRR5992854, SRR6037376, SRR6024702, SRR6012248 and SRR6008913. ("} +{"text": "We found that epigenetic signatures at enhancer and promoter elements aligns with transcriptional variation of Lct in enterocytes. Age and phenotype-specific environmental cues (lactose exposure after weaning) induced changes to epigenetic modifications and CTCF binding at select regulatory elements, which corresponded to the alterations in the intestinal Lct mRNA gradient. Thus, epigenetic modifications in combination with CTCF binding at regulatory elements account for the transcriptional gradient in Lct in cells of the same type. Epigenetic divergence within enterocytes may contribute to the functional specialization of intestinal subregions.Transcriptional variation in histologically- and genetically- identical cells is a widespread phenomenon in tissues, yet the processes conferring this heterogeneity are not well understood. To identify contributing factors, we analyzed epigenetic profiles associated with the Within a tissue, seemingly identical cells have been found to exhibit a large degree of variation in their transcriptomes145781in vitro cultures found significant heterogeneity2021in vivo and explored its potential in regulating same cell type transcriptional gradients in tissue. To answer this question, we examined transcriptional and epigenetic variation of the lactase gene (Lct) in enterocytes along the proximal-to-distal axis of the mouse small intestine. Lct, which is responsible for lactose metabolism, represents an ideal model to identify DNA modifications that can contribute to transcriptional heterogeneity for tissue subregion specialization, as it exhibits a distinguished transcriptional gradient in enterocytes; a gradual elevation from duodenum to jejunum, followed by a steady decline to the ileum2324Lct mRNA expression is dependent upon DNA modifications at key genomic regulatory elements18Lct transcription along the length of the intestine remains uninvestigated. In addition to this, we also examined how aging and the environment can modify Lct expression through DNA modifications at Lct genomic regulatory elements.Epigenetic mechanisms could contribute to transcriptional variation within the same cell type, as epigenetic modifications are important in regulating gene transcription17Lct. Aging and environmental exposures (lactose feeding after weaning) resulted in DNA modification changes at these regulatory sites and changes in Lct transcriptional patterns. Overall, DNA modification patterns support the aging- and environmentally-induced gradients of Lct mRNA and, more broadly, could affect phenotypic outcome by modifying transcriptional programs within same cell types.Here, we found that in enterocytes isolated from different intestinal regions, DNA modifications at specific genomic regulatory elements were concordant with the transcriptional variation of Lct transcriptional patterns along the proximal-distal axis of the small intestine, which enables intestinal subregion specialization in lactose metabolism mouse small intestine; the proximal, middle and distal segments of the duodenum (segment 1\u20132), jejunum (segment 3\u20135), and ileum (segment 6\u20139), and observed a gradient in steady-state Lct mRNA levels , and then gradually declined toward the proximal duodenum (p\u2009=\u20090.008 compared to segment 3) and distal ileum (p\u2009=\u20091.2\u2009\u00d7\u200910\u221210). In contrast, Mcm6 mRNA levels did not change across the intestinal segments. This indicates that enterocytes from the mid-duodenum to mid-jejunum are most specialized for the digestion of lactose.Enterocytes exhibit differing tabolism . We examLct and its neighbouring gene, Mcm6. We included Mcm6 in our investigation because DNA variation in this gene, particularly at MCM6 intron 13, affects inter-individual differences in lactase levels in humans2829Lct mRNA levels . DNA modification densities in this region were lowest in segment 3 (34.7%) and reached maximal levels in the distal ileum , an active enhancer at Lct intron 2 (H3K4me1 & H3K27ac), and an active promoter at Lct exon 1 (H3K4me3 & H3K27ac). Mcm6 exon 13\u2013intron 13, Lct exon 1 and intron 2 showed binding of CTCF, a protein that facilitates interactions between transcription regulatory sequences by affecting chromatin architecture32Lct exon 1 and intron 2 overlapped GATA1 binding sites. GATA1 is a member of GATA transcription factor family that modulates Lct expressionLct contained chromatin signatures of enhancers and a promoter, along with an enrichment in transcriptional regulatory proteins.We next examined ENCODE data of adult mouse small intestineelements . We founLct mRNA levels in enterocytes across the small intestine of infant mice (P6) and adult mice (P60). Lct mRNA levels in infants were mostly similar across intestinal segments, and were 5\u201315 fold higher than adults differed substantially in Lct abundance, and consistently, DNA modifications differed significantly between adult segments , while there was an 18% increase segment 1 (p\u2009=\u20091.3\u2009\u00d7\u200910\u22124) and a 12% increase in segment 3 or lactose-free milk (lac\u2212) for 60 days. In response to lactose treatment, Lct mRNA levels increased in the enterocytes of distal intestinal segments and 8 by 1.3- to 2- fold (p\u2009=\u20090.023) compared to mice in the lactose-free group could modify transcriptional patterns and induce epigenetic changes in enterocytes along the small intestine. For this experiment, we fed mice either lactose-containing milk or showed no statistically significant change . Distal intestinal segments exhibited a significant decrease in CpG modification densities at Lct exon 12 , Lct intron 8 , and Lct intron 2 . Segment differences in CTCF binding were most apparent at Mcm6 exon 13\u2013intron 13, and to a lesser extent at Lct intron 2. In infant mice, on the other hand, we found that CTCF was absent from Lct\u2013Mcm6 regulatory elements 28Lct transcription in vitro4243Mcm6 exon 13\u2013intron 13 is important to Lct regulation in both mice and humans262845Mcm6 exon 13\u2013intron 13 site was not sensitive to environmental cues (i.e. milk), indicative that age-dependent epigenetic programming of Mcm6 exon 13\u2013intron 13 is not malleable. By contrast, other regulatory elements affecting the Lct gradient (i.e. Lct intron 2 and intron 8) remained epigenetically adaptive to environmental signals, enabling a partial recovery of Lct expression in the adult intestine after weaning. Therefore, localized DNA modification changes accumulating with age may facilitate the intestinal Lct transcriptional gradient, while some remain partially dynamic to environmental signals.Our results indicate that divergent epigenetic programming enables aging- and environmentally-induced changes in gene transcription occurring in cells of the same type . In partLct transcriptional gradient along the small intestine, and inversely correlated to DNA modification profiles at these genomic regulatory elements, particularly at Mcm6 exon 13\u2013intron 13. CTCF is a multifunctional protein that participates in many epigenetic regulatory functions, including insulation via enhancer blocking, imprinting, X chromosome inactivation, and both transcriptional activation and repression473749Lct\u2013Mcm6 locus functions as an intestinal region-specific transcriptional activator in adult mice, potentially by enabling DNA looping of distal enhancers, such as the Mcm6 exon 13\u2013intron 13 locusLct through unobstructed accumulation of DNA modifications and epigenetic silencing. On the other hand, the lack of CTCF at Lct\u2013Mcm6 sites in infant mice signifies that CTCF binding is not required for the expression of high Lct mRNA per se. Rather, CTCF occupancy (and its putative effects on chromatin structure) occurs in tandem with DNA modification changes following weaning, to prevent uniform epigenetic silencing of Lct with age. The resulting effect in adulthood is that CTCF and the opposing DNA modification landscape work in concert to facilitate a transcriptional gradient in Lct across the intestine.CTCF occupancy was concordant with the Lct253551525DNA modifications and CTCF may work in tandem with transcription factors to create and maintain age-dependent transcriptional gradients in cells of the same type. Transcription factors can interact with gene enhancers, including those affecting Profiling epigenetic modifications of individual cell types in a tissue region-specific manner could offer insights into tissue specialization. Our findings emphasize the future studies should examine epigenetic contributions to the transcriptional divergence of numerous genes within cells of the same type, as these could help explain why tissue subregions can perform diverse biological functions41054ad libitum sterile food and either 2% lactose-containing milk or lactose-free milk in the place of water for 60 days. All animal procedures were approved by the Institutional Animal Care Committee of the Toronto Centre for Phenogenomics (TCP) and compiled per the requirements of the Canadian Council on Animal Care and Province of Ontario Animals for Research Act.Infant C57BL/6NCrl mice at postnatal day 6 (P6) and adult mice at postnatal day 60 (P60) and 90 (P90) were used to investigate epigenetic changes across the small intestine (segments 1\u20139). To investigate diet/environmental associated epigenetic changes, P30 mice were supplied Lct mRNA levels were examined in each of the nine intestinal segments. For each segment, a small proximal portion (~30\u2009mg) was homogenized with a ceramic bead-based homogenizer. Total RNA was extracted using Qiagen RNeasy Mini Kit with Qiagen RNase-free DNase I. RNA yield was quantified using NanoDrop ND-1000 (Thermo Fisher Scientific), and RNA integrity was verified via the Agilent Bioanalyzer 2100 system (Agilent Technologies). Purified RNA was converted to cDNA using High Capacity RNA-to-cDNA Kit (Life Technologies). Lct (Mm01285112_m1) and Mcm6 (Mm00484848_m1) mRNA levels were quantified using TaqMan Gene Expression Master Mix (Life Technologies) using Applied Biosystems ViiA 7 real-time PCR system. The enterocyte marker Villin-1 (Mm00494146_m1) mRNA was used as endogenous control for both Lct and Mcm6 gene expression. \u0394\u0394Ct was used to calculate the relative steady-state mRNA levels of each sample. Analysis was performed using repeated-measures (RM) ANOVA, and significant interactions were analyzed by Tukey\u2019s honest significant difference (HSD) post hoc comparisons. Mcm6 mRNA levels did not display tissue subregion variations, indicating that within cell-type transcriptional gradients can be gene specific.Enterocyte Villin-1 promoter . Significance was set at p\u2009<\u20090.01 after Bonferroni correction for multiple testing. Clusters of significantly associated modified cytosines contained 3 or more cytosines within 500\u2009bp. P-values are expressed as the \u2013log p-value of the correlation coefficient, with the sign (\u2009+\u2009/\u2212) representing the direction of Pearson\u2019s correlation (SLP). Genomic sites with DNA modification clusters significantly associated with Lct transcriptional variation were further investigated. At these sites, significant changes in the average % DNA modification status in enterocytes across the intestine was determined by one-way ANOVA. Mcm6 mRNA and lac\u2212 P90 mice. A group-wise variance matrix showed that heavily modified CpGs (>90%) lacked deviation and these were removed from analysis in this experiment . We determined which genomic sites showed significant age-associated differences by analyzing change in DNA modification (adults P60 minus infants P6). Diet-associated DNA modification changes were identified by comparing LACperiment . SignifiLct\u2013Mcm6 loci and two negative control locations up/downstream of this site (primers listed in Chromatin immunoprecipitation (ChIP) was performed to investigate CTCF binding in intestinal segments 1, 3, 5, 7 and 9 of both P6 and P60 mice (n\u2009=\u20093 per group). Tissue homogenization and ChIP were performed using the MAGnify ChIP kit (Life Technologies). Immunoprecipitation was performed overnight, using 3\u2009\u03bcl monoclonal CTCF antibody (Pierce G.758.4), and negative control reactions used 1\u2009\u03bcg of mouse IgG antibody (Life Technologies). Input controls were also taken for each sample. qPCR was performed with Universal SYBR Green Supermix (Bio-Rad) in triplicate for four Accession codes: Bisulfite sequencing data can be access on GSE76373.How to cite this article: Oh, E. et al. Transcriptional heterogeneity in the lactase gene within cell-type is linked to the epigenome. Sci. Rep.7, 41843; doi: 10.1038/srep41843 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations."} +{"text": "Left atrial volume (LAV) is an important prognostic predictor in cardiac disease. LAV is not routinely evaluated by cardiac magnetic resonance (CMR) as acquisition of a full volume dataset is time consuming, and previous authors have shown calculation of LAV using the biplane area-length method (BAL) from routinely acquired 4 and 2 chamber views significantly underestimates true volume. We hypothesized this underestimation was due to standard CMR 4CV and 2CV images foreshortening the atrium, and that additional 4CV and 2CV images piloted from mid mitral valve to the mid posterior wall of the left atrium (LA piloting) would enable rapid, accurate calculation of LAV using BAL.We evaluated 3-D datasets from 44 consecutive patients undergoing retrospective 320 slice cardiac computed tomographic studies. True 3-D left atrial volume (gold standard) was calculated at end systole by a blinded observer excluding pulmonary veins and left atrial appendage. A second blinded observer manipulated images to create standard \u2018CMR' 4 and 2 chamber views piloted from mid mitral valve to LV apex (standard LV piloted) enabling measurement of LAV using BAL. The dataset was then manipulated / 're-piloted' from mid mitral valve to the middle of posterior LA (LA piloted) and LAV was re-measured - see figure.As previously shown, LAVI calculated with BAL from LV piloted 4CV and 2CV images significantly underestimates true LAV (see table). Mean LAV calculated from LA piloted images was not significantly different from true LA volume and there was a strong correlation between the 2 with narrow confidence intervals.Accurate calculation of LAV can be made using BAL method from LA piloted images, and is superior to calculation from standard LV piloted images. Addition of two LA piloted images to a standard CMR protocol may enable rapid and accurate calculation of an important prognostic marker for cardiovascular disease."} +{"text": "We aim to evaluate the prevalence of vitamin D deficiency in patients with systemic lupus erythematosus (SLE) and investigate the association between total, free and bioavailable vitamin D serum concentrations and disease activity. Patients with SLE (ACR 1997) consecutively seen at UNIFESP\u2019s outpatient\u2019s clinics had disease activity measured after clinical and laboratory evaluation using SLEDAI (Systemic Lupus Erythematosus Disease Activity Index). 25-hydroxyvitamin D (25(OH)D) serum concentrations measured by chemiluminescence and vitamin D binding protein (DBP) measured by ELISA were used to calculate free and bioavailable vitamin D. Healthy blood donors were used as controls. A total of 142 patients (71.4%) had 25(OH)D serum concentrations below 30 ng/mL. Total 25(OH)D serum concentration was associated with disease activity categorized in 5 continuous groups of SLEDAI. 25(OH)D serum concentrations were higher among patients with SLEDAI 1\u20135 and lower in those with severe activity (SLEDAI\u226520) (p <0.05). On the other hand, no statistically significant difference was observed for DBP, free and bioavailable vitamin D measurements in the disease activity subgroups evaluated. Vitamin D deficiency is highly prevalent among patients with SLE and was associated with higher disease activity. DBP serum level and calculation of free and bioavailable vitamin D were not associated with SLE disease activity. Systemic lupus erythematosus (SLE) is a chronic multisystem inflammatory autoimmune disease . Severalin vitro evidence implies that vitamin D modulates the differentiation and activity of T and B-lymphocytes and, therefore, the production of autoantibodies [Vitamin D deficiency seems to be associated with immunological abnormalities in SLE. Some tibodies . On the tibodies , the litAn association between high disease activity in SLE with low vitamin D serum concentrations has been reported, but these results are controversial . In chilstatus than the total 25(OH)D serum concentration measurement. In the present study we assess the prevalence of vitamin D deficiency in a cohort of patients with SLE and examine the association between total, free and bioavailable vitamin D serum measurements with disease activity.Part of the disagreement regarding a potential role for vitamin D in SLE disease activity ,12,15,13The study included a total of 199 patients diagnosed with SLE according to the American College of Rheumatology (ACR) 1997 classification criteria . All parPatients with overlapping findings with other systemic autoimmune diseases, rituximab use or plasmapheresis six months before or during the course of the study, bone marrow transplantation, acquired immunodeficiency syndrome (AIDS), neoplastic disease and common variable immunodeficiency were excluded. Use of vitamin D supplements was not an exclusion criterion.All patients were contacted for clarification on the nature of the study and gave written informed consent to participate in the study. Minors/children were not included in the present study. The UNIFESP\u2019s Ethics Committee approved the study protocol.Anthropometric, demographic and clinical data collected from the electronic medical charts or clinical interview included age, sex, race, weight, height, disease duration and medication in use. Disease activity was measured using Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) at the time that blood samples were collected. Nephritis was assessed taking into account the presence of the following parameters: urine protein-to-creatinine ratio (or 24-hour urine protein) representing 500 mg protein/24 hours or red blood cell casts or renal biopsy with abnormalities suggestive of lupus nephritis.Potential differences in vitamin D serum concentrations in this sample of patients with SLE were examined using the categorization of disease activity in 5 distinct levels: Inactive disease (SLEDAI = 0), light activity (SLEDAI 1 to 5), moderate activity (SLEDAI 6 to 10), high activity (SLEDAI 11 to 19) and severe activity (\u226520 SLEDAI), as previously published . These cA total of 150 healthy volunteers selected among blood donors without SLE were used as control group.Blood samples were collected by intravenous puncture and serum aliquots were stored at -80\u00b0C for the biochemical analyses described below.25(OH)D serum measurements were performed on a Siemens ADVIA Centaur apparatus using chemiluminescence technique. Total 25(OH)D measurement coefficient of variation using this method is 11.7%.Vitamin D binding protein (DBP) was measured in SLE patients by monoclonal antibody ELISA (Cloud-Clone Corp. kit\u2014USCN Life Science Inc.) using standard technique. The coefficient of variation (CV%) for DBP measurement is 4.8%. DBP measurements in the study were considered only when duplicate measures were available. In 39 SLE samples DBP was not performed in duplicate and so was not used in the analysis.DBP measurements were used to calculate free and bioavailable vitamin D according to previously developed equation further Bone densitometry data was available for a total of 107 patients included in the present analysis. Bone mineral density (BMD) measurements were performed at the lumbar spine (L1-L4) and proximal femur using dual energy X-ray absorptiometry (DXA) . The coefficient of variation for BMD measurement was 1.5% and 2% at lumbar spine and total hip, respectively.t test for normally distributed variables and Mann-Whitney test for variables with non-normal distribution. For prospective analyses (dependent samples), quantitative variables were compared using ANOVA. Categorical variables were analyzed using Chi-square test with Bonferroni correction for multiple comparisons. Statistical analyzes were performed using SPSS software version 17.0 . Significance level was set as p <0.05.Descriptive statistics was used to characterize patients and their groups. Quantitative variables were compared between groups of independent samples using Student's A total of 199 consecutive patients with SLE were included in the study. Demographic, clinical and laboratorial data for these patients and their healthy controls are shown in Vitamin D serum concentrations according to SLEDAI categories are shown in A statistically significant difference between 25(OH)D serum concentrations was observed between the categories of disease activity differ significantly in the proportions of disease activity (p = 0.001). The proportion of patients with vitamin D sufficiency (25(OH)D \u2265 30 ng/mL) is significantly higher in the groups SLEDAI 0 and 1\u20135 when compared to 25(OH)D values below 30 ng/mL (p = 0.003). In patients with severe activity (SLEDAI \u226520) we observed the contrary: the proportion of patients with 25(OH)D serum concentration lower than 30 ng/mL was significantly higher as compared to values \u2265 30 ng/mL (p = 0.026).As shown in status and season, body mass index (BMI), Bone Mineral Density (BMD) and use of medication were also tested. There was no statistically significant difference in the mean serum concentrations of 25(OH)D (p = 0.179), free and bioavailable vitamin D (p = 0.441) between the different seasons (ANOVA) .25(OH)D serum concentrations were not significantly associated with disease duration or BMI. Both free and bioavailable vitamin D was also not associated with those variables.statuses (Spine and hip BMD did not correlate significantly with 25(OH)D, free or bioavailable vitamin D serum concentrations. Moreover, BMD did not differ significantly among the various vitamin D statuses .Possible associations between 25(OH)D serum concentration and the use of medications were considered. status of vitamin D: both free and bioavailable fractions of 25(OH)D did not differ between the various categories of SLE disease activity.In the present study we investigated the association between vitamin D serum concentration, its free and bioavailable fractions and disease activity in SLE patients. We have observed a very high prevalence of vitamin D deficiency and insufficiency among SLE patients. Only 28.6% of the patients were vitamin D sufficient (serum 25(OH)D \u2265 30 ng/mL). Vitamin D deficiency was significantly more frequent in SLE patients than in healthy controls. Vitamin D deficiency is then more prevalence in SLE patients as compared to the Brazilian general population without SLE \u201325. SignSome cross-sectional studies have shown an inverse correlation between vitamin D serum concentration and disease activity in SLE patients ,6,12. OnThe importance of free and bioavailable vitamin D has been tested in other clinical scenarios. Low 25(OH)D serum levels have been associated with high risk of multiple sclerosis . Free anIt was recently demonstrated that DBP measurements using monoclonal ELISA are biased by DBP genotype and race . DBP valVitamin D serum concentrations did not correlate with the seasons in sample of patients with SLE, unlike what has been observed in the general population . This isIn this study, vitamin D supplementation was associated with increased 25(OH)D serum concentrations. The reported doses of 400 to 1000 IU/day do not seem to be sufficient to ensure adequate levels of vitamin D. About 35% of patients using cholecalciferol still presented vitamin D deficiency or insufficiency, suggesting that higher doses are required for proper supplementation in these patients.Due to its cross-sectional nature, our study cannot establish a causal relationship between vitamin D serum concentration and disease activity in SLE patients. If low vitamin D serum concentrations are causal co-factor in the immunological disturbances that characterize SLE or if, on the contrary, the inflammatory disease process and low sun exposure causes reduction in vitamin D serum concentrations will still require further studies . It is astatus and disease activity in SLE.In conclusion, we have demonstrated a very high frequency of vitamin D deficiency and insufficiency among patients with SLE. In this clinical scenario, disease activity was associated with lower serum concentrations of 25(OH)D. DBP measurements with monoclonal ELISA and free and bioavailable vitamin D calculations did not differ among different categories of SLE disease activity. Prospective studies are needed to investigate and establish a potential causal relationship between vitamin D S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file."} +{"text": "Gene ontology analysis identified several key pathways, including lymphocyte activation. A list of differentially expressed genes is provided. The raw data was uploaded to the sequence read archive (SRA) database and the BioProject ID is PRJNA432903.Invasive ductal carcinoma is the most common type of breast cancer. Here, we provide a whole transcriptome shotgun sequencing dataset conducted with ten samples of invasive ductal carcinoma tissue and three samples of adjacent normal tissue from a single Korean breast cancer patient . Differentially expressed genes (DEGs) were identified with a false discovery rate (FDR)-adjusted Specifications TableValue of the data\u2022This RNA-seq data provides a deep sequencing of ten samples of invasive ductal carcinoma tissue and three samples of adjacent normal tissue from a Korean breast cancer patient \u2022The heterogeneous expression data from spatially distinct tumor samples can be used for various evaluation purposes.\u2022Gene ontology analysis revealed that lymphocyte activation and PPAR signaling pathway are significantly up- and down-regulated pathways, respectively, in breast cancer tissue compared to adjacent normal tissue.1p-value cutoff of 0.05. Gene ontology analysis indicated that several pathways are associated with the onset or progression of breast cancer.Total RNA was extracted from ten samples of cancer tissue and three samples of adjacent normal tissue from a Korean patient with breast cancer. RNA-seq was performed to profile transcriptomes of breast cancer and normal samples. Differentially expressed genes were identified with an FDR-adjusted 22.1One tissue sample of invasive ductal carcinoma from breast tissue and a corresponding adjacent normal tissue were biopsied from a Korean woman with informed consent. This study was approved by the institutional review board of Catholic Medical Center . The tumor and adjacent normal tissues were divided into ten and three samples, respectively. Poly(A) RNA was purified from 1\u202fg total RNA from each sample, and cDNA was synthesized using SuperScript II (Invitrogen). Sequencing libraries were prepared using the TruSeq RNA Library preparation kit (Illumina) and sequenced using HiSeq. 2500 (Illumina).2.2https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/) with Cutadapt (version 1.1.2) Sequenced reads were trimmed using Trim Galore between cancer and normal samples were identified using Cufflinks with the Cuffdiff function (version 2.2.1) samples . When th samples .Fig. 1Co2.4http://metascape.org) Gene ontology (GO) analysis was performed to identify key pathways regarding the DEGs that were identified without the C3 sample. The top 100 up-regulated (or down-regulated) DEGs that were highly expressed (> 10 average FPKM) were analyzed using Metascape ("} +{"text": "Bicuspid aortic valve (BAV) is known to be heritable and often shows a concomitant aortopathy which was thought also to be present in some family members. While one recent study reported normal ascending aortic diameters in family members of the general BAV population, little is known about aortic function in such family members. We used advanced aortic cardiovascular magnetic resonance (CMR) and peripheral blood analysis to assess evidence for an underlying aortopathy in family members of bicuspid aortic valve patients.\u00a9 as well as circulating matrix metalloproteinases (MMP) 2 and 9 from a peripheral blood sample. A subset of 10 family members also underwent comprehensive 4D flow MRI assessment.We prospectively enrolled 229 participants in total. This included 42 families (42 BAV index cases and 132 family members) in whom at least one member was affected by BAV. Participants over 6 years of age underwent CMR, and those under 6 years echocardiography. We also recruited 55 age and sex-matched healthy volunteers. Advanced aortic assessment included aortic diameters, pulse wave velocity (PWV), arterial stiffness, maximum rate of systolic dysfunction (MRSD) and distensibility by CMR, total peripheral resistance (TPR) by Vicorder2, p = 0.22).11% (14/132) family members were found to have a BAV. The remaining 118 family members with a normal functioning tricuspid aortic valve had an average age of 38.7 years with a mean pulse pressure 57 mmHg. Compared to sex, age and blood-pressure matched healthy volunteers all family members had normal sinus, ascending and descending aortic diameters. There was no difference in PWV , ascending, proximal and distal descending aortic strain . MRSD and distensibility , TPR , MMP2 and MMP9. In the subset of family members undergoing advanced 4D flow MRI assessment, there was no difference in flow angle , flow displacement and wall shear stress compared to healthy volunteers trileaflet aortic valves of patients with a bicuspid aortic valve have normal aortic size and function. These findings point towards the importance of haemodynamic factors rather than additional haemodynamic-independent mechanisms in the aortopathy of bicuspid aortic valve patients."} +{"text": "APC1311 truncating mutation orthologous to human APC1309, analysing whole samples and microdissected dysplastic epithelium. Gene set enrichment analysis revealed differential expression of gene sets similar to human normal mucosa versus T1 stage polyps. Transcriptome analysis of whole samples revealed many differentially-expressed genes reflecting immune infiltration. Analysis of microdissected dysplastic epithelium was markedly different and showed increased expression in high-grade intraepithelial neoplasia of several genes known to be involved in human CRC; and revealed possible new roles for GBP6 and PLXND1. The pig model thus facilitates analysis of CRC pathogenesis.We compared gene expression in low and high-grade intraepithelial dysplastic polyps from pigs carrying an APC) tumour suppressor initiates formation of most adenomas in the human gut, and is responsible for most cases of familial adenomatous polyposis (FAP), an inherited predisposition to colorectal cancer (CRC)1. While the severity of the condition varies considerably, FAP patients typically develop tens to hundreds of adenomatous polyps in the colon and rectum in early life and have greatly increased risk of developing CRC2.Functional disruption of the adenomatous polyposis coli , chromosomal instability, aberrant DNA methylation and histone modification3. However the events that determine whether an early stage polyp proceeds towards cancer have not yet been identified. As Sievers et al.4 recently reported, human precancerous polyps are very heterogeneous, which hinders comparative molecular analyses.Several genetic and epigenetic alterations have been implicated in this progression, including tumour suppressor genes and proto-oncogenes (APC gene (APC1311) orthologous to a human APC1309 mutation responsible for FAP5, to determine if molecular changes in the pig model parallel events in small polyps in humans, and whether pigs can be used to reveal new events in early CRC development. The value of pigs in modeling CRC and other cancers has been discussed in several publications e.g. refs APC1311 pig model has the advantage that samples can be taken from many individuals with the same initiating mutation, avoiding an important source of variation. Changes held in common across such samples are more likely to have a function in premalignant transformation.We investigated gene expression in the premalignant progression of dysplastic polyps in pigs carrying an engineered translational stop signal at codon 1311 in the APC1311 mutation, orthologous to human APC1309 responsible for a severe form of FAP5. Four generations of APC1311/+pigs have now been examined regularly by colonoscopy and high-grade intraepithelial (HG-IEN) dysplasia and carcinoma in situ or HG-IEN (n\u2009=\u20096), were analysed by next generation RNA sequencing, and transcriptome clusters specific for LG-IEN and HG-IEN polyps identified. More than 300 genes were expressed at significantly different levels between the two groups , intracellular transport , and many were immune related . Differential expression of selected genes was validated by quantitative PCR (qPCR) . AHNAK has been reported as a negative regulator of cell growth, and as a tumour suppressor that potentiates TGF\u03b2 signalling10. Inactivation of epithelial TGF\u03b2 signalling promotes development of intestinal cancer in mouse and human colon cells ex vivo12. Of those genes expressed at higher level in HG-IEN polyps, the stress-responsive gene HSPA1L (P\u2009=\u20091.7\u2009\u00d7\u200910\u22126), a member of the HSP70 family, showed the greatest difference (3.6 fold). Others had functions related to Wnt signalling (WISP1), the TNF pathway (SLC12A6), and cell apoptosis and tumour microenvironment . Pathway analysis of differentially expressed genes using PANTHER software revealed that the histamine H1 receptor mediated signalling and B cell activation pathways were significantly over-represented . Histamine receptors are known to play an important role in cancer development including CRC13. It should however be pointed out that most genes studied (n\u2009=\u2009238) were unclassified in the pathways, which is a limitation when analysing the pig genome.The progress of polyposis in 14. Equivalent data from human LG-IEN versus HG-IEN polyps was not available, precluding direct side-by-side comparison with the pig. Differential expression data for human T1 stage microsatellite stable (MSS) and microsatellite instable (MSI) polyps versus normal colonic mucosa from The Cancer Genome Atlas (TCGA) database provided the closest comparison. GSEA analysis of pig data revealed that 25 of 50 gene sets showed significant differences between LG-IEN and HG-IEN, including many sets important for tumorigenesis. Parallel analysis of the human MSS data showed 26 of 50 gene sets with q\u2009<\u20090.05, of which 19 were held in common with pig; human MSI data showed 23 of 50 gene sets with q\u2009<\u20090.05, of which 7 were held in common with pig, GSEA results are summarised in Fig.\u00a0APC mutations are frequent in non-hypermutated (MSS) colon cancers15. There was a striking degree of similarity between changes in gene sets in pig and human MSS polyps and several close parallels in key functional sets, e.g. upregulation of MYC target genes, the E2F group of genes involved in cell cycle regulation and DNA synthesis, and the cell cycle G2/M checkpoint DNA damage gene set, and downregulation of interferon alpha and gamma response targets. This was clearly only a broad comparison and there were some anomalies, such as the apparent lack of Wnt pathway upregulation T1 human polyps despite its known role in polyposis. This latter is probably explained by the heterogeneous nature of the human data and the likely high proportion of stroma in the human polyp biopsies masking the contribution of tumour cells16.To assess the relevance of our animal model to human disease, we carried out gene set enrichment analysis (GSEA)APC1311 FAP pig and in humans.These findings strongly suggest that similar molecular changes are involved in early polyp development in the 18. To investigate whether transcriptome data is useful for assessing severity of polyposis, we performed RNA sequencing on a series of histologically unclassified 1\u2009cm diameter polyps taken from 22 APC1311/+ animals. Polyp biopsies were taken from animals with very low and very high (>100 in same region) numbers of colorectal polyps. Unsupervised clustering of RNA sequencing data revealed two distinct gene expression profiles of genes with the most marked differences in expression between pig LG-IEN and HG-IEN were immune-related or known to be specific to immune cells. Immunohistochemical analysis of polyp samples confirmed infiltration of CD3+, CD4+, and CD8+ T cells Figure\u00a0. The rolet al.21 examined a limited number of cancer-related genes in microdissected human LG-IEN polyps. Our analysis of microdissected samples revealed marked differences to results from whole samples. Several genes identified as differentially expressed in the whole samples were not represented, e.g. IL7, IL17D, S100A8, S100A9, AHNAK, indicating that such transcriptional changes occurred in other tissues, such as stroma and infiltrating immune cells. New gene expression differences were also revealed by the more defined analysis. Indeed, none of the top 20 genes identified as differentially expressed in microdissected epithelium were significant in the whole sample analysis. Most notable were higher expression of PLXND1 and GBP6 in HG-IEN , p53 regulation and cell proliferation (VASH1), Wnt pathway regulation (SMARCD3), and DNA binding . Of those genes expressed at lower level in HG-IEN, the greatest differences were in RAD23B, a DNA repair enzyme, and CHST12 or immune cells (SLA2) and analysed allele expression in microdissected samples from distinct LG-IEN and HG-IEN adenoma from the same polyp is reportedly common in CRClyp Fig.\u00a0. The allAPC1311 pigs correlates with alterations in molecular pathways involved in early pathogenesis of human colorectal cancer. Microdissection and specific analysis of the epithelial component revealed that many transcriptome changes apparent in whole samples actually reflect events outside the epithelium, notably immune infiltration. Allele-specific expression was clearly evident, especially in microdissected HG-IEN, consistent with the emergence of cell subpopulations. Analysis of microdissected dysplastic epithelium also revealed possible roles for cancer-related genes not previously associated with CRC in early stage adenoma progression.In summary, pigs genetically predisposed to colorectal polyposis based on a single, well-defined genetic mutation enable systematic analysis of CRC precursor lesions, offering an important complement to the study of human patients and existing genetically-modified mouse models. Gene expression data from whole samples obtained by biopsy of polyps from Animal experiments were approved by the Government of Upper Bavaria (permit number 55.2-1-54-2532-6-13) and performed according to the German Animal Welfare Act and European Union Normative for Care and Use of Experimental Animals. All pigs were fed with normal pig diet. Blood and tissue samples were collected every 6 months.in vivo with a STORZ colonoscopy system and ex vivo with a Zeiss Stemi 11 stereomicroscope. Colonoscopy examination was carried out approximately every 6 months, starting at 3 months old. Polyp and normal mucosa biopsy samples were divided and portions were fixed as described below, frozen in 2-methylbutane (OCT) for cryosection, and snap frozen and stored at \u221280\u2009\u00b0C for molecular analyses.Macroscopic images of colonic lesions were taken 32.For histopathology analysis, specimens were fixed in 4% buffered paraformaldehyde, embedded in paraffin, sectioned (3\u2009\u03bcm) and stained with haematoxilin and eosin (H&E). Polyps were classified as LG-IEN or HG-IEN according to the AJCC TNM staging system33, embedded in paraffin and 3\u2009\u03bcm sections cut. Sections were dewaxed, microwaved , and incubated with mouse anti-porcine primary antibodies: CD3 , CD4 , CD8 . All primary antibodies were obtained from SouthernBiotech. Tissue sections were incubated with biotinylated anti-mouse IgG followed by Elite ABC kit incubation. Antibody binding was detected with a DAB Peroxidase Substrate kit (Vector Laboratories).For immunohistochemistry, specimens were fixed using the HOPE techniqueLaser microdissection (LMD) of cryosectioned samples was performed immediately after H&E staining using a Leica Microsystems Laser Microdissection Systems 6000 and Leica Application Suite software (Leica). In total, ten LMD captured crypts per polyp sample were cut, providing 300\u2013500 cells, and collected into 50\u2009\u00b5L lysis buffer from the AllPrep\u00ae DNA/RNA Micro Kit (Qiagen), and a further 50\u2009\u00b5L lysis buffer added after dissection. Dissected samples were stored at \u221280\u2009\u00b0C. Genomic DNA and total RNA from the same sample were isolated using the AllPrep\u00ae DNA/RNA Micro Kit according to the manufacturer\u2019s instructions (Qiagen).For allele expression assessment by pyrosequencing, 100\u2009pg total RNA isolated from LMD samples was used for whole transcriptome amplification using the QuantiTect Whole Trancriptome Kit according to the manufacturer\u2019s instructions (Qiagen).Libraries for RNA sequencing of whole biopsies were prepared using the TruSeq Stranded mRNA LT Kit (Illumina). RNA integrity and fragment size were tested using the RNA6000 Nano kit (Agilent) on an Agilent Bioanalyzer 2100 (Agilent).Libraries for RNA sequencing of LMD samples were prepared using SMART-Seq. 2 Ultra-low kit (Clontech) and Nextera XT DNA SMP preparation kit according to the manufacturers protocol (Illumina). RNA and dscDNA integrity and fragment size were tested using RNA6000 Pico and High Sensitivity DNA kits (Agilent).STAR aligner with default parameters34. 80% of these reads uniquely hit to the reference genome. Duplicate reads were denoted with the MarkDuplicates tool of Picard (http://broadinstitute.github.io/picard). Aligned reads (omitting duplicate reads) were assigned to gene sequences as defined in the 10.2.77 porcine gene set and counted with featureCounts35. Normalisation of read counts and estimation of fold change was carried out using DESeq. 236. Hierarchical clusters and heat maps for 60 genes with the most significantly different levels of expression were generated using the python (http://www.python.org) packages spatial and cluster of scipy37 along with matplotlib38.Libraries were sequenced with a HiSeq. 2500 ultra-high-throughput sequencing system (Illumina) to produce 100-base-paired end reads. An average of 45 million reads per sample were generated and mapped to the porcine reference genome (Sscrofa10.2) using the 2-pass method of the STAR alignments was performed according to GATK39 best practice recommendations for RNAseq41. The GATK tool SplitNCigarReads was used to split reads into exons and remove false variants resulting from overhangs. This step included reassignment of the STAR alignment mapping qualities. GATK recalibration of base scores was based on the Ensembl release 83 variant database. Variant calling was carried out using GATK HaplotypeCaller with the dontUsedSoftClippedBases option. GATK VariantFiltration was applied to clusters of at least 3 SNPs within a window of 35 bases between them with the following parameters: Fisher strand value (FS) >30.0 and a quality by depth value (QD) <2.0. The probability of allelic imbalance for each SNP was calculated based on the number of reference and alternate allele reads in heterozygous animals using a two-sided binomial test. P values were adjusted for false discovery rate to take account of multiple testing.Variant calling based on TCGAanalyze_DEA and TCGAanalyze_LevelTab. For porcine colon polyps, the numbers of mapped reads to each gene were counted using the featureCount tool implemented in the subread package (version 1.5.0-p2)35. The gene annotation file was downloaded from Ensembl (Ensembl release 86). Log2 fold change of gene expression (HG-IEN versus LG-IEN) was analysed using the R/Bioconductor package DESeq. 2 (version 1.14.0)36.Gene expression data (HTSeq-counts) for stage I human colon cancers (n\u2009=\u200980) and normal samples of colonic mucosa (n\u2009=\u200941) were retrieved from The Cancer Genome Atlas (TCGA) using the R/Bioconductor package TCGAbiolinks (version 2.2.3). From the TCGA, we also retrieved gene expression data (HTSeq-counts) for stage I microsatellite stability (MSS) (n\u2009=\u200923) and stage I low level microsatellite instability (MSI-L) (n\u2009=\u200911) human colon cancers. Log2 fold change of gene expression was analysed using the included functions 42 was performed using Human Genome Organization (HUGO) gene symbols together with log2 fold change of gene expression as input. The tool GSEAPreranked implemented in GSEA (version 2.2.3) was used to estimate gene set enrichment with the following parameters: classic enrichment statistics, 1000 permutations, and hallmark gene sets collection from Molecular Signatures Database (version 5.2)43. P values were adjusted for false discovery rate to take account of multiple testing.Gene set enrichment analysis44 was used to perform functional pathway analysis for the top >300 differentially expressed genes with PANTHER pathways as annotation data set and statistical overrepresentation test with default settings.PANTHER Gene List AnalysisPPIA and RPS28 reference genes. Reference genes were selected from a set of eight examined genes ATP6, RPS28, GAPDH, HPRT1, RPS23, TPT1, PPIA, RN18S based on stability values from the NormFinder.Total RNA was extracted using Direct-zol RNA Mini Prep Kit (Zymo Research) according to the manufacturer\u2019s instructions. 200\u2009ng total RNA was used to synthesise complementary DNA (cDNA) using Superscript IV reverse transcriptase (Invitrogen). Two-step qPCR experiments were performed using Fast SybrGreen MasterMix (Applied Biosystems) and run on an ABI 7500 thermocycler (Applied Biosystems). Primer specificity and capture temperature were determined by melt curve analysis. The relative expression difference between the groups in all tissues was calculated for each sample (\u0394\u0394CT). All cDNA samples were assayed in triplicate and relative expression levels normalised to Genomic DNA was extracted using the GenElute Mammalian Genomic DNA Kit (Sigma Aldrich) and used for allele quantification in heterozygous SNPs. The allele proportion of 50:50 on DNA level for valid pyrosequencing assays was expected. Total RNA was extracted using the Direct-zol RNA Mini Prep Kit (Zymo Research) according to the manufacturer\u2019s instructions. 200\u2009ng total RNA was used to synthesise complementary DNA (cDNA) using Superscript IV reverse transcriptase (Invitrogen). cDNA was amplified by RT-PCR, and allele expression for selected SNPs was analysed by pyrosequencing using the Pyromark Q24 system (Qiagen). Pyrosequencing primers were designed using the PyroMark Assay Design Software 2.0 (Qiagen). Allele expression differences at individual SNPs were evaluated using Students t-test.The authors will provide all primer sequences used in this study on request.Supplementary information"} +{"text": "Kinomics identified 7 peptides with increased tyrosine phosphorylation in metastases that were significantly altered (p<0.005). Based on these peptides, bioinformatics analyses identified several candidate kinases activated in metastases compared to primary tumors. The highest ranked upstream kinase was Focal Adhesion Kinase 1 (FAK1). RCC lines demonstrate evidence of elevated FAK1 activation relative to non-transformed renal epithelial cells. Pharmacologic inhibition of FAK1 with GSK2256098 suppresses in vitro tumor phenotypes. In turn, FAK1 knockdown in RCC cells suppresses both in vitro phenotypes and in vivo tumor growth. Collectively, these data demonstrate functional activation of FAK1 in metastases and provide preclinical rationale for targeting this kinase in the setting of advanced ccRCC.The introduction of targeted therapies has caused a paradigm shift in the treatment of metastatic clear cell (cc)-renal cell carcinoma (RCC). We hypothesized that determining differential kinase activity between primary and metastatic tumor sites may identify critical drivers of progression and relevant therapeutic targets in metastatic disease. Kinomic profiling was performed on primary tumor and metastatic tumor deposits utilizing a peptide substrate microarray to detect relative tyrosine phosphorylation activity. Pharmacologic and genetic loss of function experiments were used to assess the biologic significance of the top scoring kinase on Despite multiple targeted agents in the therapeutic armamentarium for clear cell (cc)-renal cell carcinoma (RCC), this disease remains largely incurable. The median overall survival (OS) is approximately 2 to 2.5 years when employing vascular endothelial growth factor (VEGF) inhibitors , 2. SecoContinuing to investigate the role of novel kinase inhibitors of alternate targets may be reasonable. Indeed, multiple kinases are likely responsible for progression and metastasis and the discovery of novel kinases may offer the opportunity to therapeutically target them with kinase inhibitors . MoreovePreliminary studies demonstrate the feasibility of kinomics profiling of primary renal tumors . HoweverAmong 96 available fresh frozen ccRCC tumor samples, 92 met quality control criteria for kinase activity. ccRCC tumor was available from 80 primary tumors and 12 metastases . The perWe next performed algorithmic analyses of these 7 phosphopeptides as a means to infer candidate upstream kinase activated in metastasis relative to primary RCC and RCC cells Figure . All linWe measured the change in the proliferative rates of RCC cells in response to GSK2256098. 786-O and RXF393 cells treated with GSK2256098 demonstrated reduced proliferation relative to untreated cells Figure . To assein vitro phenotypes, we wanted to validate the effects of FAK1 inhibition via genetic loss of function experiments. We used lentivirus to stably knockdown FAK1 expression via shRNA. Immunoblotting after puromycin selection demonstrated reduced FAK1 protein expression in cells transduced with two non-overlapping shRNA constructs relative to control vector (PLKO) transduced cells of tyrosine kinases on peptide substrates which is likely more physiologically relevant than kinase expression . We studWhen examining the panel of increased substrate phosphorylation in metastases in the overall population, FGFR1 demonstrated the highest fold difference compared to primary tumors. In addition to VEGF and PDGF, the fibroblast growth factor (FGF) pathway mediates angiogenesis and may be especially critical in mediating resistance to VEGF inhibitors \u201333. Notain vitro ccRCC cell line proliferation and FAK1 knockdown in ccRCC cells suppressed both in vitro and in vivo tumor growth. Preclinical data exist to support the induction of anoikis in RCC by targeting FAK survival signaling by quinazoline compounds . shRNAs were co-transfected into 293T cells together with packaging plasmids by following the manufacturer's protocol . RXF 393 and 786-O cells were passaged and plated in a 6-well plate and allowed to adhere for 24 h before infection. RXF 393 and 786-O cells were transduced in the presence of polybrene overnight. After 24 h cells were selected by treating with media containing 2 \u03bcg/ml puromycin.Foxn1nu ; Charles River Laboratories) following the Institutional Animal Care and Use Committee (IACUC) protocols at UAB. Five animals were injected per group and assessed over time for the development of tumors. Animals were sacrificed at 8 weeks post-injection and tumors were harvested from the flanks.786-O with PLKO.1 empty vector or FAK1 shRNA, were grown and maintained in complete media containing puromycin. Two million cells were collected and resuspended in 150\u03bcL of media and mixed with an equal volume of BD Matrigel\u2122 Basement Membrane Matrix (BD Biosciences). These cells were injected into the flanks of athymic nude mice (NU(NCr)-www.r-project.org), and the commercial MetaCore knowledge base, to develop pathway maps and biological networks. Peptides with increased phosphorylation were queried based on their phosphorylatable residues (up to 6 per peptide) on www.phosphonet.ca (Kinexus). Two scoring algorithms, \u2018V2\u2032 and \u2018Proximity\u2019, were used to identify putative upstream kinases responsible for peptide phosphorylation. The \u2018score\u2019 is the combination of both V2 and Proximity scores as previously described [The degree of phosphorylation on each PamChip peptide probe was measured kinetically using Evolve software (PamGene), that measured FITC labeled anti-phosphotyrosine antibody binding to each phosphorylated peptide substrate during the 60 min assay and were further analyzed using BioNavigator software (PamGene), the open source microarray statistical package, R (escribed , 18. In escribed , 18. Forescribed , 18. All"} +{"text": "However, unfavorable effects occur relative to controls when Cpt1bm-/- mice are fed a 25% fat diet, including decreased activity and fat free mass and increased intramuscular lipid and serum myoglobin. In this study we explore if a low fat, high carbohydrate diet can ablate the unfavorable effects while maintaining the favorable phenotype in Cpt1bm-/- mice. Mice were fed either 10% fat (low fat) or 25% fat (chow) diet. Body composition was measured biweekly and indirect calorimetry was performed. Low fat diet abolishes the decreased activity, fat, and fat free mass seen in Cpt1bm-/- mice fed chow diet. Low fat diet also reduces serum myoglobin levels in Cpt1bm-/- mice and diminishes differences in IGF-1 seen between Cpt1bm-/- mice and control mice fed chow diet. Glucose tolerance tests reveal that glucose clearance is improved in Cpt1bm-/- mice relative to controls regardless of diet, and serum analysis shows increased levels of muscle derived FGF21. Electron microscopic analyses and measurements of mRNA transcripts show increased intramuscular lipids, FGF21, mitochondrial and oxidative capacity markers regardless of diet. The favorable metabolic phenotype of Cpt1bm-/- mice therefore remains consistent regardless of diet; and a combination of a low fat diet and pharmacological inhibition of CPT1b may offer remedies to reduce blood glucose.Inhibiting fatty acid oxidation is one approach to lowering glucose levels in diabetes. Skeletal muscle specific Carnitine Palmitoyltransferase 1b knockout mice (Cpt1b Considerable evidence supports the idea that oversupply of dietary fat exceeds the storage capacity of adipose tissue and leads to ectopic lipid accumulation resulting in \u201cmetabolic stress\u201d in skeletal muscle, liver, pancreas and possibly other tissues, contributing to insulin resistance \u20134. One pCarnitine palmitoyltransferase 1 (Cpt1) catalyzes a rate limiting step of fatty acid oxidation via the shuttling of long chain fatty acids across the mitochondrial membrane. The enzyme is expressed as an isozyme subset, with Cpt1b being the predominant form expressed in skeletal muscle, heart, and brown adipose tissue , 4. Cpt1While these studies implicate Cpt1b as a potential target for further investigation and development of drugs used to treat type II diabetes mellitus, other studies have demonstrated detrimental side effects in their administration. Widespread inhibition of all Cpt1 isoforms by etomoxir can result in steatohepatitis . OxfenicUsing a skeletal muscle-specific CPT1b KO model, we showed that mitochondrial FAO inhibition results in a myriad of diabetes risk factors without inducing insulin resistance . Perhapsm-/- mice [m-/- mice, side effects such as the accumulation of intramyocelluar lipids (IMCL), myodegeneration, and decreased activity and energy expenditure also occur [More recently we showed that inhibition of mitochondrial FAO induces FGF21 expression specifically in skeletal muscle and that FGF21 acts in a paracrine manner to increase glucose uptake under low insulin conditions and has no effect on adiposity in Cpt1b-/- mice . Though so occur .m-/- mice might show fewer negative side effects associated with excessive fat intake when fed a diet low in fat and high in carbohydrates. In the current report we examine the effects of varying diets upon Cpt1bm-/- mice in order to determine the flexibility of their previously observed phenotypes; and to assess optimal diet conditions in systems where Cpt1b action is decreased in an attempt to maintain or even restore insulin sensitivity.As Cpt1b continues to develop as an intriguing target for the treatment of metabolic disorders, it is important to consider possible effects of varying dietary intake during such treatment. Known clinical variants of Cpt1b have not been functionally characterized as yet. However, maintenance strategies for individuals with Cpt1a defects involve limiting exercise, fasting, and fat intake, while also ensuring high dietary levels of carbohydrates . We therCpt1b to Mlc1f-Cre transgenic mice for specific ablation of Cpt1b in skeletal muscle, but not cardiac tissue [Cpt1b targeting construct and breeding scheme to generate constitutive muscle-specific inactivation of Cpt1b are shown in In order to address the role of impaired skeletal muscle FAO in the development of insulin resistance, we targeted the muscle-specific isoform of carnitine palmitoyltransferase, Cpt1b. We crossed mice bearing floxed alleles of c tissue . C57L/B62 asphyxiation followed by cervical dislocation, according to approved procedures of the Panel on Euthanasia of the American Veterinary Medical Association.Animal studies were conducted at Pennington Biomedical Research Center\u2019s AALAC-approved facility. All experiments were in compliance with the NIH Guide for the Care and Use of Laboratory Animals, and approved by the Pennington Biomedical Research Center Institutional Animal Care and Use Committee (IACUC). All mice were 3\u20134 month old male mice on C57BL/6 background unless specified otherwise. Unless otherwise stated, mice were multi-housed, and all were exposed daily to 12 hours of light and 12 hours of dark and fed different diets as indicated . Mice weBody composition was measured using a Bruker NMR Minispec (Bruker Corporation). Serum and plasma collections were performed by submandibular bleeding. GTT were performed following a 4-h fast by intraperitoneal injection of 20% D-glucose (40mg of glucose per mouse). Behavioral and indirect calorimetry studies were done in a 16-chamber Oxymax system (Columbus Instruments) as described previously , 16. ForELISA kits were used for measurement of IGF-1, Myoglobin, and FGF21 in serum. IGF-1 and Myoglobin were tested in the fed state, while FGF21 was assayed in overnight fasted serum.T procedure described previously [Total RNA from red quadriceps muscle was isolated for later qRT-PCR using an RNeasy Mini Kit supplemented with DNase digestion (Qiagen). cDNA was then synthesized with an iScript cDNA synthesis kit and was used for qRT-PCR with the SYBR Green system (Bio-Rad). Analysis was conducted using the \u2206\u2206Ceviously . QuantifSoleus muscles were fixed in 2% (wt/vol) glutaraldehyde/1% (wt/vol) formaldehyde and then postfixed in 2% (wt/vol) osmium tetroxide, en bloc stained in 0.5% uranyl acetate, and embedded in Epon-NMA. 70nm ultrathin sections were mounted on collodion-coated copper grids, stained with Reynolds lead citrate, and imaged with a JEOL 100CX transmission electron microscope at the Socolofsky Microscopy Center at Louisiana State University.t-tests where normality was established using GraphPad Prism software and the D'Agostino-Pearson normality test. P\u2264 0.05 was considered significant. For adiposity, gene expression, and food intake studies populations were not often normally distributed or had sample sizes of N<8. For these analyses GraphPad Prism software and Mann-Whitney U Tests were performed as a measure of significant differences with P\u22640.05 considered significant. JMP software from SAS was used for ANCOVA analysis.Data are expressed as mean \u00b1 s.e.m. For measurement of serum proteins and lipids, blood glucose levels, and activity Microsoft Excel software was used for analysis of variance with paired two-tailed Student\u2019s m-/- mice and control (Cpt1bfl/fl) mice could be ablated by varying nutrient composition within diets, mice were fed low fat (10% fat) or chow (25% fat) diet diet . As fat at) diet . Howeverdecrease . Becausem-/- animals are due to differences in fat mass, which begins to deviate several weeks before differences in food intake become significant , but are lower than Cpt1bm-/- mice fed a chow diet with marginal significance (p = 0.06). Cpt1bm-/- mice on a low fat, high carbohydrate diet therefore show a partial correction of the muscle damage observed in Cpt1bm-/- mice eating a chow diet.An additional negative characteristic of Cpt1bcontrols . Similarnificant . Circulam-/- mice have reduced activity relative to control mice when fed the 25% fat chow diet [m-/- mice have similar levels of activity when fed a 10% fat diet compared to controls when fed chow diet [m-/- mice relative to controls on chow diet . Much like the serum hormones and activity levels discussed above, the differences seen between genotypes in these lipid metabolites is diminished during low fat feeding. The changes in the levels of these biomarkers and behavioral patterns together with the abolition of weight gain differences suggest that many of the negative physiological effects upon Cpt1bm-/- mice fed chow diet are restored to control levels with low fat diet feeding.In addition to muscle damage, we have previously shown that Cpt1bhow diet . Howeverfat diet . Cpt1bm-how diet . This ishow diet . Though m-/- mice have improved glucose clearance relative to control animals when fed chow diet [m-/- mice at baseline and throughout the time course of glucose tolerance tests in mice fed either chow or low fat diet Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S1 FigANCOVA analysis of weight (A) and fat mass (B) as a function of covariables genotype and average daily Kcal intake. White circles indicate controls and black circles indicate Cpt1bm-/- mice .(DOCX)Click here for additional data file.S2 Fig.(DOCX)Click here for additional data file."} +{"text": "Inhibitors of FGF receptor signaling are currently entering the clinic and our results suggests that FGF receptor signaling is a therapeutic target in cancers with TE fusion gene expression and PTEN loss.Prostate cancer is the most common visceral malignancy and the second leading cause of cancer deaths in US men. Correlative studies in human prostate cancers reveal a frequent association of the TMPRSS2/ERG (TE) fusion gene with loss of PTEN and studies in mouse models reveal that ERG expression and PTEN loss synergistically promote prostate cancer progression. To determine the mechanism by which ERG overexpression and PTEN loss leads to transformation, we overexpressed the TE fusion gene and knocked down PTEN in an immortalized but non-transformed prostate epithelial cell line. We show that ERG overexpression in combination with PTEN loss can transform these immortalized but non-tumorigenic cells, while either alteration alone was not sufficient to fully transform these cells. Expression microarray analysis revealed extensive changes in gene expression in cells expressing the TE fusion with loss of PTEN. Among these gene expression changes was increased expression of multiple FGF ligands and receptors. We show that activation of fibroblast growth factor receptor signaling plays a key role in transformation induced by TE fusion gene expression in association with PTEN loss. In addition, Prostate cancer (PCa) is the most common malignancy in American men, affecting one in nine men over 65 years of age . CurrentCorrelative studies in human PCa reveal a frequent association of the TE fusion gene with loss of PTEN and studies in mouse models reveal that ERG expression and PTEN loss synergistically promote PCa progression \u201318. ConsFibroblast growth factors (FGFs) are a family of 18 different ligands that bind with variable affinity to four different FGF receptors (FGFR1-4). There iTo determine the mechanism by which TE fusion gene expression and PTEN loss leads to transformation, we expressed The TE fusion gene and/or knocked down PTEN in an immortalized but non-transformed prostate epithelial cell line. We show that TE expression in combination with PTEN loss can transform these immortalized but non-tumorigenic cells while either alteration alone was not sufficient to fully transform these cells. Furthermore, we show that activation of fibroblast growth factor receptor (FGFR) signaling plays a key role in transformation induced by ERG expression in association with PTEN loss. In addition, we show that PTEN loss results in widespread but variable increases in FGF ligands and receptors in PCa.To determine if TE fusion gene expression and PTEN loss were sufficient to transform prostate epithelial cells we constructed cell lines with expression of the TMPRSS2/ERG fusion gene (TE), stable knockdown of PTEN with shRNA (PTEN KD) or both alterations (PTEN KD/TE) using the PNT1A cell line. The PNT1A cell line was originally established by SV40 T-Ag immortalization of benign prostate epithelial cells . PNT1A ain vitro phenotypes of the four cell lines. The PTEN KD, TE and PTEN KD/TE cell lines all grew significantly faster in vitro than control cells although PTEN KD cells grew slower than both the TE and PTEN KD/TE cells and SV40-T antigen (to confirm origin from PNT1A) and were negative for AR were identified. As shown in Figure Examination of the genes that were uniquely altered at the chosen thresholds in PTEN KD/TE showed that both FGF3 and FGFR4 were upregulated in these cells. Given that there is a large body of evidence that FGFs and their receptors are upregulated in PCa and that FGFR signaling is a potential drug target , we chosWe then examined both FGFR1 and FGFR4 phosphorylation the PTEN KD/TE cell lines compared to PNT1A controls after stimulation with FGF2 using phospho-FGFR ELISAs. As seen in Figure To determine whether FGFR signaling plays an important role in invasion induced by TE expression and/or PTEN KD we used the FGFR kinase inhibitor AZD4547. We also used the AKT kinase inhibitor AZD5363 to confirm that the cellular phenotypes induced by PTEN knockdown are sensitive to inhibition of this key downstream target activated by PTEN loss. In control PNT1A cells there was a small but not statistically significant inhibition by either inhibitor alone but the combination inhibited invasion approximately 20% Figure . AZD4547We then evaluated the impact of treatments single or in combination on soft agar colony formation of PTEN KD/TE cells. AZD4547 alone and in combination with AZD5363 almost completely abolished soft agar colony formation Figure . AZD5363We then examined whether PTEN knockdown altered FGF signaling in PCa cell lines. We analyzed RNAs from 22RV1 cells (which have wild-type PTEN) and 22RV1 with PTEN knockdown using a human FGF pathways Q-RT-PCR array (Thermo Fisher) to identify changes in FGF ligands and/or receptors induced by PTEN knockdown, with increases of 2-fold or decreases 0.5-fold considered biologically significant. As shown in Figure To determine whether there is evidence of an association of PTEN loss with expression of FGF signaling components in human PCa specimens we examined mRNA expression of human FGF ligands and FGF receptors in 131 primary cancers with mRNA with from the Taylor dataset in cBiopExamination of the smaller number of metastatic cases with mRNA revealed a similar pattern. As seen in Figure We also carried out Gene Set Enrichment Analysis using the FGF ligands and FGFRs described above on the Taylor dataset of primary and metastatic PCa. We found a highly significant enrichment of FGF ligands and FGFRs in cases with PTEN loss . VCaP and HEK 293T cells were cultured in DMEM with 10% FBS. PNT1A cells were obtained from the European Type Culture Collection. LNCaP, VCaP and 293T cells were obtained from the American Type Culture Collection. Cells were obtained between 2001 and 2016, expanded, frozen and stored as stocks in liquid nitrogen. All cell lines are authenticated by STR analysis at MD Anderson Cancer Center Characterized Cell Line Core Facility.5\u2019-AATGTTTGGATAAATATAG-3\u2019 (Clone Id:V2LHS_92314) and 5\u2019-TAATAATACACATAGCGCC-3\u2019 (V2LHS_92317). Lentiviral shRNA was produced by cotransfection of the Trans-Lentiviral packaging mix with a shRNA transfer vector into 293T packaging cells.GIPZ shRNA clones targeting human PTEN gene and non-silencing pGIPZ control vector, both containing the Turbo GFP reporter, puromycin-resistance gene, and elements required to allow packaging of the expression construct into virions were purchased from the Cell-Based Assay Screening Core Facility at Baylor College of Medicine. The mature antisense sequences are: NotI and ligated into pcDH-CMV-EF1-neomycin vector. TE lentiviral particles were generated in 293T cells using pCDH-CMV-MCSEF1- TE3+72-neo and pPACK Lentivector packaging kit (System Biosciences).The TE isoform (3+72) lentivirus was consin vitro experiments. V2LHS_92317 were used for the in vivo studies because of their better PTEN knockdown efficiency. Similarly, PNT1A with TE expression were generated by infecting with TE lentivirus and selected in 200ug/mL G418. The combined PTEN KD/TE stable cell line was generated by infection with equal amounts of TE and PTEN shRNA lentivirus and selected with puromycin and G418. The cell lines were generated twice and had the same in vitro phenotypes both times.To make stable cell lines, RPMI medium supplemented with lentiviral supernatants PTEN KD, TE or both were supplemented with 5 \u03bcg/mL polybrene and incubated with cells for 24 hours. The PTEN KD cells were maintained in medium with 400ug/mL puromycin. PNT1A stably transfected with PTEN shRNA with clone Id V2LHS_92314 and V2LHS_92317 were used for To evaluate the role of PTEN in controlling FGF and FGFR receptor expression a VCAP cell with stable PTEN knockdown was constructed as described for the PNT1A stable cell lines using PTEN ShRNA lentivirus (V2LHS_92317) and vector control. Q-RT-PCR showed a 80% knockdown efficiency. For 22RV1 cells PTEN was knocked down transiently using PTEN siRNA purchased from Sigma; PTEN sense: 5\u2019-AAC CCA CCA CAG CUA GAA CUU dTdT-3\u2019 and antisense: 5\u2019-AAG UUC UAG CUG UGG UGG GUU dTdT-3 and scramble siRNA control. Transfections were performed using 50 nM with RNAiMax transfection reagent (Invitrogen) for 48 hours and RNA was extracted. PTEN mRNA knockdown was 75% compared to controls. To evaluate FGF23 expression LNCaP or VCaP, cells were treated for 48 hours with 300 nM AZD5363 or vehicle, RNA was extracted and used for Q-RT-PCR of FGF23 mRNA.Total RNA was isolated using the RNeasy Mini Kit (Qiagen). 500 ng total RNA were used for cDNA reverse transcription using amfi Rivert Platinum cDNA Synthesis Enzyme Mix (GenDEPOT). 1-5 uL of cDNA was used for Q-RT-CR using in a final reaction volume of 15ul using SYBR Green PCR Master Mix (Applied Biosystems) for PTEN or TaqMan Fast Advanced Master Mix (Applied Biosystems). The PTEN primers were sense: 5\u2019- AGCGTGCAGATA ATGACAAGG-3\u2019 and antisense: 5\u2019- TGGATCAGA GTCAGTGGTGTC-3\u2019, with an annealing temperature of 60\u00b0 C. FGFR1 and FGFR4 were analyzed using Taqman MGB probe (FAM-FGFR1 and FGFR4 and VIC-ACT). FGF23 primers and conditions were used as published in previously . Q-RT-PCCells were lysed in modified RIPA buffer (Santa Cruz) supplemented with PMSF, protease inhibitor and sodium orthovanadate. Protein concentration of the lysates were determined using BCA protein assay reagent (Bio-Rad); 30\u03bcg of the extracted protein was mixed with Laemmli sample buffer containing \u03b2-ME, denatured, separated using 10% PAGEr Gold Precast Gels (Lonza), and transferred using iBlot Gel Transfer system (Invitrogen) and iBlot Gel Transfer Stacks Nitrocellulose (Invitrogen). Anti-ERG , anti-PTEN , anti-\u03b2-actin antibody (Santa Cruz Biotech), antibodies were used at 1:1000 dilution with blocking with 5% skim milk. After incubation with primary antibodies overnight at 4\u00b0C, horseradish peroxidase\u2013labeled secondary antibodies were then applied to the membranes for 1 h at room temperature. Signals were visualized using ECL Western blotting detection substrate (Thermo Scientific).5 cells were seeded on 24-well plates in triplicate and attached cells were counted using Beckman Cell counter. The experiment was repeated three times. Final experiments were also confirmed by MTT assay using CellTiter 96 Aqueous One Solution Cell Proliferation Assay (Promega).1\u00d710Invasion assays were conducted using pre-coated BD Matrigel Invasion Chamber 24 well plates (BD Biosciences) in triplicate. A total of 5000 cells were plated per insert and after 48 hrs culture, non-invading cells in the upper chambers were removed and the invading cells on the lower surface of the membrane were fixed and stained with DAPI. The membranes were mounted on slides, photographed under fluorescent microscopy at 10X and cells enumerated using Image J software. Each experiment was repeated 3 times. For drug treatment AZD4547 or AZD5363 was added in media in the bottom wells with the final concentration of 300 nM for both drugs as described previously .Five thousand cells expressing were mixed with the 0.7% agarose (top agar) with warm 2\u00d7RPMI 1640 + 20% fetal bovine serum and plated in each well of a 6-well plate on top of the prepared 1% base agar. Plates were incubated at 37\u00b0C for 3 weeks before the foci were stained with 0.005% Crystal Violet and counted. For drug treatment, AZD4547 or AZD5363 was supplemented to the 2XRPMI media for the preparation of the base agar and top agar with the final concentration of 300 nM. Twice a week 100 uL of RPMI medium supplemented with 300 nM of one or both drugs was added to the top of the agar to keep the agar humid.Immunohistochemistry (IHC) was carried out using the general procedures described previously , using sPhospho-FGFR1 levels from the cells were measured using PathScan\u00ae Phospho-FGF Receptor 1 (panTyr) Sandwich ELISA Kit according to manufacturer's protocol. The cell lysates were prepared using cell lysis buffer included in the kit supplemented with 1mM PMSF, sonicated, spun down at 4 \u00b0C and supernatant protein concentration was measured using BCA method (Bio-Rad.). A total of 300 ug protein was loaded to the ELISA strip in duplicate in total volume of 200 uL adjusted with sample buffer included in the kit. After incubations and washes following the manufacturers protocol optical density at 450 nM was measured using a microplate reader . The positive control was 30 ug of A201 treated with FGF and 200 ug of 22RV1 cells lysate treated with 300 nM AZD4547 was served as negative control. Similarly, Phospho-FGFR4 (PanTyr) ELISA kit was used for Phospho-FGFR4 detection in cell lysates.6 PNT1A TE, PNT1A-PTEN KD, PNT1A PTEN KD/TE in or PNT1A vector control cells in SCID mice and after 3 months the genitourinary tracts of all mice were harvested and H&E sections evaluated for tumor formation by a pathologist (MI). In a 2nd experiment, SCID mice were injected subcutaneously over both lateral flanks with 3\u00d7106 cells of each genotype in 200ul volume mixed with 100ul Matrigel (BD Bioscience). After 3 months, any palpable tumors were collected fixed and paraffin-embedded for H&E examination to confirm the presence of cancer cells.All procedures were approved by the Baylor College of Medicine Institutional Animal Use and Care Committee. In the first experiment, eight-week old male SCID mice were injected intraprostatically with 2\u00d710Total RNA were extracted with RNeasy Mini Kit (Qiagen), according to the manufacturer's recommendations. The cDNA reverse transcription and fluorescent labeling reactions were carried out using Invitrogen SuperScript Plus Direct cDNA Labeling System with Alexa Fluor S\u2019-Aminohexylacrylamido-dUTP. 60K Whole Human Genome Oligo Microarray chip (Agilent Technologies) using SureHyb DNA Microarray Hybridization Chambers as described previously .For each treatment group, top differentially expressed genes relative to control were defined , and the set of top differential genes found for any treatment group were clustered, using a supervised approach as described elsewhere . ExpressTaqMan\u2122 Array Human FGF Pathway , which contains 92 assays to FGF pathway associated genes and 4 assays to candidate endogenous control genes, was used to evaluate FGF ligands and/or receptor changes in 22RV1 and 22RV1 with PTEN knockdown or VCaP and VCaP with PTEN knockdown cell lines. 25ng of cDNA was added into each well of these 96 wells FGF arrays along with TaqMan\u00ae Gene Expression Master Mix (2x) to make the total reaction of 10ul. Real-time PCR were performed using Thermo Fisher StepOnePlus\u2122 by standard protocol.Numerical values were compared using t-test. Proportions were compared with chi-squared test or Fishers exact test. Differences were considered significant if p<.05.Bioinformatics analysis: Yiqun Zhang and Chad J. Creighton.Study supervision and data analysis: Michael Ittmann."} +{"text": "The postnatal heart undergoes highly coordinated developmental processes culminating in the complex physiologic properties of the adult heart. The molecular mechanisms of postnatal heart development remain largely unexplored despite their important clinical implications. To gain an integrated view of the dynamic changes in gene expression during postnatal heart development at the organ level, time-series transcriptome analyses of the postnatal hearts of neonatal through adult mice were performed using a newly developed bioinformatics pipeline. We identified functional gene clusters by principal component analysis with self-organizing map clustering which revealed organized, discrete gene expression patterns corresponding to biological functions associated with the neonatal, juvenile and adult stages of postnatal heart development. Using weighted gene co-expression network analysis with bootstrap inference for each of these functional gene clusters, highly robust hub genes were identified which likely play key roles in regulating expression of co-expressed, functionally linked genes. Additionally, motivated by the role of the transcription factor Sox6 in the functional maturation of skeletal muscle, the role of Sox6 in the postnatal maturation of cardiac muscle was investigated. Differentially expressed transcriptome analyses between Sox6 knockout (KO) and control hearts uncovered significant upregulation of genes involved in cell proliferation at postnatal day 7 (P7) in the Sox6 KO heart. This result was validated by detecting mitotically active cells in the P7 Sox6 KO heart. The current report provides a framework for the complex molecular processes of postnatal heart development, thus enabling systematic dissection of the developmental regression observed in the stressed and failing adult heart. Mef2c) , while 2 genes were consistently downregulated in Sox6 KO heart [leucine carboxyl methyltransferase 1 (Lcmt1) and nudix-type motif 6 (Nudt6)]. Among these, Myocd is known to be involved in cardiovascular development [Myocd also plays an important role in the functional maturation of the postnatal heart. Although the functions of the rest of the genes in heart development are as yet unknown, our result indicates that they are Sox6 target candidates, thus could be involved in the maturation of the postnatal heart as well.In our analysis of DE genes between the Sox6 KO and control hearts, the majority of genes showed differential expression only at a single time point, while some genes demonstrated consistent differential expression at multiple time points . Of theselopment as well elopment ; therefoEgr1, a hub gene in cluster J4, showed clearly different expression patterns between cardiac ventricle tissues and isolated cardiomyocytes A box plot showing normalized microarray data from triplicate samples at each developmental stage. (B) A multidimensional scaling (MDS) plot demonstrating similarity between each genotype and each developmental stage. Each axis represents an arbitrary unit and is therefore dimensionless.(TIF)Click here for additional data file.S2 Fig(A) PCA with SOM clustering of gene expression. The expression profile of each gene is represented, and genes that belong to different clusters are indicated by different colors and separated by PC1 and PC2. (B) Loading plots for PC1 and PC2 showing contributions of each variable to the primary components. (C) Expression profiles of each SOM cluster. Scaled expression along developmental stages is shown in box plots together with cluster categories and names . Of note, the SOM clusters obtained with mean intensity values were comparable but slightly different from those with median intensity values , suggest(TIF)Click here for additional data file.S3 FigGenes were sorted according to the average ranking calculated from TOM-based connectivities after bootstrap inference , and plotted against standard deviation (s.d.) of the ranks. Each dot represents individual gene. Genes ranked in top 50% in each SOM cluster are shown. Red dots: hub genes (ranked in top 5%). Black dots: other genes.(TIF)Click here for additional data file.S4 FigWorkflow for analyzing control and Sox6 KO mice is shown. Data and results are shown in red, and processing steps are shown in green.(TIF)Click here for additional data file.S5 FigSox6 mRNA levels were quantified by RT-qPCR. Data are normalized for the reference genes (see the n = 3).Total RNA was extracted from control and Sox6 KO mice ventricles, and (TIF)Click here for additional data file.S6 Figp-values (highlighted) obtained by AME.Significant enrichment of the Sox6 binding motif is shown as adjusted (TIF)Click here for additional data file.S7 FigThe number of probes for DE genes was counted at each time point and is shown as bar graphs. DE genes unique to each time point are shown in dark gray, and DE genes at multiple time points are shown in color (i.e. the same color indicates the same set of genes). (A) Upregulated genes in the Sox6 KO heart. (B) Downregulated genes in the Sox6 KO heart.(TIF)Click here for additional data file.S8 FigEno1 from cluster N1, Egr1 from cluster J4, and Lyz1 from cluster A4) obtained by this study and O\u2019Meara et al. (2015) are exemplified Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 TableGenes were sorted according to the average ranking calculated from TOM-based connectivities after bootstrap inference . Red: hub genes Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 TableData from this study and O\u2019Meara et al. (2015) (\u201c1A_FPKM\u201d sheet of Online Table I) are merged using gene symbol, and mean intensity values/FPKM values and standard deviations are shown.(XLSX)Click here for additional data file."} +{"text": "IRF6) gene variants are significant genetic contributors to orofacial clefts. Although approximately three hundred IRF6 gene variants have been documented, their effects on protein functions remain difficult to interpret. Here, we demonstrate the protein functions of human IRF6 missense gene variants could be rapidly assessed in detail by their abilities to rescue the irf6-/- phenotype in zebrafish through variant mRNA microinjections at the one-cell stage. The results revealed many missense variants previously predicted by traditional statistical and computational tools to be loss-of-function and pathogenic retained partial or full protein function and rescued the zebrafish irf6-/- periderm rupture phenotype. Through mRNA dosage titration and analysis of the Exome Aggregation Consortium (ExAC) database, IRF6 missense variants were grouped by their abilities to rescue at various dosages into three functional categories: wild type function, reduced function, and complete loss-of-function. This sensitive and specific biological assay was able to address the nuanced functional significances of IRF6 missense gene variants and overcome many limitations faced by current statistical and computational tools in assigning variant protein function and pathogenicity. Furthermore, it unlocked the possibility for characterizing yet undiscovered human IRF6 missense gene variants from orofacial cleft patients, and illustrated a generalizable functional genomics paradigm in personalized medicine.Large-scale sequencing efforts have captured a rapidly growing catalogue of genetic variations. However, the accurate establishment of gene variant pathogenicity remains a central challenge in translating personal genomics information to clinical decisions. Interferon Regulatory Factor 6 ( IRF6, a gene of established importance in orofacial cleft pathogenesis. IRF6 functions are well-conserved across species, and allowed us to test the functions of human IRF6 variants by their abilities to rescue zebrafish embryos depleted of irf6. Many variants previously labeled pathogenic and loss-of-function retained full wild type-level protein activity, suggesting they could potentially represent benign, rather than disease-causing genetic variations. This paradigm is applicable to other genes and will allow researchers to rapidly triage gene variants for further study and clinicians to provide more accurate genetic diagnoses.Advances in sequencing technologies have led to rapid increases in personalized genetics information. Millions of differences exist when comparing the genomes of two individuals, accounting for the diversity of humans and occasionally disease pathogenicity. Accurate determination of the functional consequences of these differences is critical for translating personal genetic information into clinical decision. Various methods have been devised to meet this challenge. However, they are imperfect, especially in their abilities to interpret rare variants. Rare variants must be evaluated by multi-pronged approaches to establish disease pathogenicity, one crucial approach being functional testing through experimental models. Here, we utilized a zebrafish model to rapidly evaluate the protein functions of rare gene variants of The rapid development of next-generation sequencing technologies has ushered in a new era of personalized medicine for a myriad of diseases . Large-sIRF6, ENSG00000117595) [IRF6 are among the most common genetic causes of cleft lip and/or palate (CL/P) [IRF6 gene sequence is highly conserved across vertebrates and contains two functional domains, a helix-turn-helix DNA-binding domain and a SMIR/IAD protein-binding domain [Irf6 led to a CL/P phenotype in addition to oral epithelial adhesions, poor epithelial barrier functions, and improper skin stratification, revealing a potentially important role for the oral epithelium in directing palate development [IRF6 gene variants have been catalogued [IRF6 gene variants remain significant challenges. Moreover, different computational programs use algorithms that weight various aspects of amino acid change differently and often provide conflicting predictions on protein function for the same missense mutation [One well-studied example of a common congenital malformation associated with gene variants is that of orofacial clefts associated with the transcription factor Interferon Regulatory Factor 6 0117595) \u201315. Orof0117595) . Pathoge595 \u201315. e (CL/P) . The IRFg domain ,18. Fromelopment ,20. Apprmutation .IRF6 gene variants and its well-understood biology make IRF6 an ideal model to examine the challenges of assigning variant protein function and pathogenicity. In order to develop a rapid functional assay to determine the protein functions of a large number of IRF6 gene variants, we utilized a novel irf6 null zebrafish model by taking advantage of the finding that maternally-deposited irf6 transcripts are required for the proper development of the embryonic epithelium (periderm) during epiboly [irf6 null model, we were able to establish maternal-null irf6-/- homozygotes where 100% of the embryos lacked Irf6 and ruptured unless rescued by functional Irf6 protein. This irf6 rescue assay was used to test the protein functions of human IRF6 missense gene variants, and provide an additional line of biological evidence to help bridge the gap between gene variant identification and pathogenicity assignment.The large number of human epiboly . By usinirf6 null model and investigate the role of irf6 in periderm and craniofacial development, CRISPR-Cas9 was used for mutagenesis targeted at exon 6 of irf6 at the start of the SMIR/IAD protein-binding domain (irf6 coding region (NC_007133.7(NM_200598.2):c.772_779del), here forward referred to as \u201c\u03948bp\u201d (irf6+/\u03948bp) were in-crossed to produce wild type, heterozygous and homozygous progeny at the expected Mendelian ratios with normal embryonic and craniofacial development and homozygous (\u03948bp/\u03948bp) irf6 embryos that developed normally up to the sphere stage (4 hours post fertilization = hpf). However, shortly thereafter these maternal irf6\u03948bp/\u03948bp embryos, regardless of genotype, failed to appropriately initiate epiboly compared to wild type embryos, resulting in the separation of the animal pole structures from the underlying yolk, periderm rupture, and embryonic lethality in 100% of embryos between 5\u20136 hpf compared to the complete wild type 492 amino acids (55 kD) from permutations of wild type and maternal/paternal homozygous irf6\u03948bp/\u03948bp crosses. Transcript levels of irf6 were undetectable in all maternal irf6\u03948bp/\u03948bp crosses at 4 hpf, regardless of whether the embryos were homozygous or heterozygous for the \u03948bp irf6 allele .The 8bp deletion in the (55 kD) . It has tivities . Thus, i6 allele . Conversmparable . Western crosses . Furtherhenotype . Taken tirf6 \u03948bp allele was predicted in silico to have no off-target sites in coding regions of the zebrafish genome (crispr.mit.edu), we sought to determine the specificity of the embryonic periderm rupture phenotype from CRISPR irf6 gene disruption by injecting wild type zebrafish irf6 mRNA into maternal-null irf6-/- embryos at the one-cell stage, and assessing whether the periderm rupture phenotype can be rescued. Injection of zebrafish irf6 mRNA reliably rescued the rupture phenotype in maternal-null irf6-/- embryos, and the rescued embryos were able to initiate epiboly and undergo normal embryonic development indistinguishable from wild type embryos (Although the CRISPR gRNA used to generate the embryos .IRF6 across species ranging from human to zebrafish [IRF6 to retain its function in zebrafish. Wild type human IRF6 cDNA (NM_006147.3) was isolated, in vitro transcribed into mRNA, and injected into maternal-null irf6-/- zebrafish embryos to determine whether the periderm rupture phenotype could be rescued. The ability of human IRF6 mRNA to fully rescue the maternal-null irf6-/- periderm rupture phenotype was indistinguishable from that of zebrafish irf6 mRNA. In addition, maternal-null irf6-/- embryos rescued by either zebrafish or human IRF6 mRNA appeared indistinguishable from wild type embryos throughout embryonic development with normal craniofacial morphologies both programs agree the variant disrupts protein function, resulting in a loss-of-function protein, 2) the programs disagree on the effects of the variant on protein function, and 3) both programs agree the variant does not disrupt protein function. Human IRF6 missense gene variants were then mapped to their corresponding nucleotides in the zebrafish irf6 cDNA by sequence conservation, in vitro transcribed into mRNA, and microinjected into maternal-null irf6-/- zebrafish embryos to assess their ability to rescue the periderm rupture phenotype (irf6-/- rupture phenotype (IRF6 gene variants were grouped according to their abilities to rescue periderm rupture and mapped to the predicted human IRF6 protein structure (protein-binding domain and C-terminus) generated by ExPASy, the distribution of the amino acid residues suggested variants that could not rescue mostly resided in protein secondary structures and thus likely to disrupt protein conformation and function was performed in strict accordance with protocols approved by Massachusetts General Hospital IACUC (Protocol 2010N000106).Danio rerio were maintained in accordance with approved institutional protocols at Massachusetts General Hospital, as described [Zebrafish escribed . Embryosescribed by hoursirf6 exon 6 gRNA was generated by in vitro transcription from a T7 promoter as described [cas9 mRNA was synthesized from a T3 promoter using the mMESSAGE mMACHINE T3 transcription kit (Ambion) and purified using the RNeasy mini kit (Qiagen). One-cell staged zebrafish embryos were injected directly in the cytoplasm with 2 nl of a solution containing 25 ng/\u03bcl of gRNA and 100 ng/\u03bcl of cas9 mRNA. Genomic DNA for genotyping was isolated from either whole 24 hpf embryos or tail fin clips using the HotSHOT method [Potential CRISPR gRNA target sites were identified using the CRISPR design program at (zifit.partners.org/ZiFiT/ and crispr.mit.edu). The escribed . Zebrafiescribed was lineT method . GenotypStage-matched zebrafish embryos were flash-frozen in liquid nitrogen and homogenized with a micropestle in TRIzol (Invitrogen). Total RNA was isolated using phenol-chloroform exaction and digested using DNase I (Ambion) to remove genomic DNA contamination. Total RNA was quantified using a NanoDrop spectrophotometer and 5 \u03bcg was used for reverse transcription using the SuperScript III cDNA synthesis kit (Invitrogen). Quantitative RT-PCR was performed using PowerUP SYBR Green qPCR master mix (Invitrogen) on the StepOne Plus RT-PCR platform (Applied Biosystems). Elongation factor 2\u03b1 or \u03b2-actin were used as internal controls for expression normalization. Amplification specificity was checked using melt-curve analysis. Control amplifications were performed on samples either without reverse transcription or template.Zebrafish embryos were enzymatically dechorionated with pronase (Sigma) and deyolked according to supplemeirf6 (NM_200598.2) and human IRF6 (NM_006147.3) cDNA was synthesized using GeneArt (Invitrogen) and sub-cloned into the pCS2+8 vector (Addgene #34931) at the EcoRV site in the MCS. IRF6 gene variants were identified in previously published literature and mapped to their corresponding nucleotides in the zebrafish irf6 cDNA. PCR-based site-directed mutagenesis was performed to generate pCS2+8 vectors containing irf6 missense gene variants using the Q5 site-directed mutagenesis kit (New England Biolabs) with amplification primers designed by the NEBaseChanger online tool (http://nebasechanger.neb.com). Plasmids containing irf6 missense gene variants were isolated using the QIAprep spin miniprep kit (Qiagen) and sequence verified with either complete plasmid sequencing or Sanger sequencing through the entire cDNA region.Full-length zebrafish IRF6 missense gene variant cDNAs were linearized by NotI-HF and purified using QIAquick PCR purification columns (Qiagen). Variant mRNA was synthesized by in vitro transcription using the SP6 mMESSAGE mMACHINE transcription kit (Ambion) and linearized plasmids as template. cDNA template was digested using DNase I and variant mRNA was purified using the RNeasy mini kit (Qiagen) and quantified with either a NanoDrop spectrophotometer or Qubit fluorometer (Invitrogen). Variant mRNAs were diluted to 800 ng/\u03bcl and stored as aliquots in -80\u00b0C. For embryo microinjections, 2 nl of the injection mix was delivered directly into the cytoplasm of one-cell staged embryos with variant mRNA diluted to a final concentration of 50 ng/\u03bcl.Plasmids (pCS2+8 backbone) containing individual Zebrafish embryos were fixed in 4% paraformaldehyde (PFA) at 4\u00b0C overnight and subsequently transferred into 100% methanol prior to whole-mount in situ hybridization (WISH). WISH and DIG-labeled riboprobe synthesis were performed essentially as described in . All rib2O2) until pigmentation of cells were no longer present. Acid-free alcian blue staining was performed essentially as described [Zebrafish embryos were fixed in 4% PFA at 4\u00b0C overnight and bleached and SIFT (http://sift.jcvi.org) for computational predictions of missense variant protein function. The ExPASy SWISS-MODEL online tool (https://swissmodel.expasy.org) was used to align the IRF6 amino acid sequence with the known crystalline structure of IRF1 to model the IRF6 SMIR/IAD protein-binding domain and C-terminus. The resulting protein structures were visualized using PyMOL. For statistical analysis of experimental data, error bars represent \u00b12x standard mean error (SEM), and statistical significance was interrogated using two-tailed Student\u2019s T-tests with <0.05 as the P-value cut-off. For the identification of IRF6 missense variants from the ExAC database (exac.broadinstitute.org) and gnomAD database (gnomad.broadinstitute.org), IRF6 was queried and the results were sorted for missense gene variants.Human terature and ente"} +{"text": "The World Health Organization (WHO) international standard for hepatitis A virus (HAV) RNA nucleic acid assays was characterized by complete genome sequencing. The entire coding sequence and noncoding regions were assigned HAV genotype IB. This information will aid the design, development, and evaluation of HAV RNA amplification assays. Hepatovirus genus within the Picornaviridae family that causes acute liver infection, potentially resulting in HAV disease (Hepatitis A virus (HAV) is a\u00a0nonenveloped single-stranded RNA virus\u00a0with a genome of 7.5\u2009kb and is a member of the disease . World H disease . ElucidaThe first WHO IS developed for HAV RNA genome amplification techniques (HAV IS 00/562) represents a cornerstone of HAV diagnostic algorithms (Taq HiFi (Invitrogen) using primer pairs described previously of an HAV-positive plasma donation sourced from the National Plasma Repository, South African National Blood Service, in 2011. To elucidate sequence identity, viral RNA was extracted using total nucleic acid isolation (TNAi) extraction methodologies on an Ampliprep instrument (Roche Ltd.). Rescued RNA was reverse transcribed and directly amplified with the SuperScript III one-step reverse transcription-PCR (RT-PCR) system with Platinum eviously . In combHAV-specific amplicons were sequenced using a cycling profile of 25 cycles on a 3130XL sequencer (ABI Ltd.) in forward and reverse directions and assembled and analyzed using Geneious v7 software. A contiguous sequence was represented by 7,386\u2009nt, with a total GC content of 38.2%. Phylogenetic analyses indicated that the 3rd HAV IS clusters among HAV genotype IB sequences, with no subtype recombination events identified. The complete genome sequence reported here, derived from the 3rd WHO HAV RNA IS, will support genome amplification assay development for HAV RNA detection and quantification in environmental samples and clinical studies.KY003229.The genome sequence of the 3rd WHO IS for HAV RNA reported here has been deposited in GenBank under accession number"} +{"text": "These data reflect the perceptions of beach water quality drawn from a convenience sample of 238 visitors to Georgia (USA) beaches collected in June\u2013July 2017 and are related to the research article entitled \u201cWater quality and the perception of risk: a study of Georgia, USA, beachgoers\u201d These data may thus be useful to researchers comparing coastal perceptions between residents and visitors.1This dataset presents information on the varied perceptions of what constitutes water quality including whether respondents view the absence of waterborne pathogens as the primary feature of water quality at tourist beaches. Data also include which illnesses associated with waterborne pathogens are perceived as linked to unclean water by respondents. 2Researchers collected these data using a quantitative survey offered either online via recruiting on Facebook or, primarily, through in-person collection of paper surveys from visitors to Georgia beaches in June\u2013July 2017. In-person data collection focused on two major Georgia recreational beaches (Tybee Island and Jekyll Island) during peak swimming season. This study was approved by the Georgia Southern University Institutional Review Board with participants\u2019 consent required to complete the survey. Data were analyzed using IBM SPSS 23 and ArcMap 10.4.1 .\u2022Consent .\u2022Description of which factor most represent clean water to a respondent.\u20225 variables asking whether a respondent perceives as not (0) or having (1) as association with polluted water.\u2022Age.\u2022Sex.\u2022Hispanic ethnicity.\u2022Race .\u2022Race2 (dichotomized to white (0) or minority (1)).\u2022Income (dichotomized as income below U.S. median household income (0) or higher (1)).\u2022Education (dichotomized as without a 4-year college degree (0) or with one (1)).Data variables include:"} +{"text": "Our earlier results show a decreased peak circumferential . SLV and SRV patients were asymptomatic at time of imaging and were post total cavo pulmonary connection (TCPC).Acquisition Protocol:4 images: \u03b5cc; and b) Fast-Strain Encoded (fSENC)5 images: \u03b5L.Strain information was acquired at three short axis slices at basal , and apical (uncoupled) locations in all 18 subjects in a 1.5T MRI scanner (Philips Acheiva) using: a) Complementary Spatial Modulation of Magnetization (CSPAMM)Data Analysis:cc and \u03b5L across all cardiac phases were calculated from SAX slices using DiagnosoftTM. Global, free-wall and septal strain were calculated at both locations and ventricular coupling index (VCI) is calculated as (\u03b5cc * \u03b5L /100).\u03b5Strain values of SLV and SRV subjects demonstrate significant differences compared to normal subjects. Figure 2)Strain at the septal location is significantly reduced in single ventricle patients while the freewall strain is relatively normal.3)Circumferential strain of the SV progressively reduces from the apex to the base, while the longitudinal strain increases.4)VCI is significantly reduced at the basal septum - pointing to the deleterious effect of the ventricular coupling between the systemic ventricle and the dysfunctional ventricle. Myocardial fiber arrangement and the hypoplastic chamber likely affect the regional differences demonstrated in this study Figure .Longitudinal strain is increased in regions where the circumferential strain is decreased in SV patients. However at the basal septum, where the dysfunctional ventricle is attached to the septum, it appears that the systemic ventricle mechanics is affected by deleterious ventricular coupling. Further studies are needed to understand differences between SRV and SLV cardiac biomechanics."} +{"text": "We investigated possible vaccinia virus (VACV) in urban house cats in Brazil. Serum samples from 6 cats were positive for VACV by PCR, indicating likely VACV circulation among house cats in urban areas of Brazil. This finding highlights the importance of epidemiologic surveillance to avoid outbreaks among urban human populations. Some cats had clinical illness inconsistent with orthopoxvirus infection, which can overlap with other common dermatologic diseases affecting cats of the cats were female. Thirteen cats (4.7%) had outdoor access, and 37 (13.4%) were admitted to the veterinary clinic for ing cats Table. Ming cats Figure 1ing cats Table. Aing cats Figure 1\u2013We describe evidence of VACV circulation in cats in an urban environment in Brazil. Many studies have attempted to elucidate VACV outbreaks and risk factors in rural and wild areas \u2013 Figure 1Hydrochoerus hydrochaeris) from the Pampulha region tested positive for VACV gene detected in domestic cats compared with homologous sequences of several other orthopoxviruses, Belo Horizonte, Brazil, September 2012\u2013December 2014."} +{"text": "During a 10 year study period 234 patients were admitted on 371 occasions with a total of 566 acute varicealbleeding episodes. Of these, 173 patients had 343 variceal bleeds which required balloon tamponade toachieve initial control of bleeding during 229 admissions and were then referred for emergency injectionsclerotherapy. Sixty-eight percent of these patients had alcoholic cirrhosis and 42% were poor risk GradeC patients. Injection sclerotherapy was performed initially using the rigid Negus oesophagoscope undergeneral anaesthesia and later using the fibreoptic endoscope under light sedation. Definitive control ofvariceal bleeding was achieved with sclerotherapy during 197 hospital admissions (92%). Of the 17failures of emergency sclerotherapy, 4 patients died from uncontrolled bleeding and 13 patientsunderwent major surgical intervention. Definitive control of variceal bleeding was achieved with a singleinjection treatment in 138 hospital admissions (70%). Complications were mostly of a minor nature andoccurred at a rate of 6% per injection treatment. The overall hospital admission mortality was 36%. Themajority of patients died due to liver failure. The mortality in patients who required 4 injection treatmentsto control variceal bleeding was 71%. Injection sclerotherapy is proposed as the emergency treatment ofchoice for patients whose variceal bleeding continues or recurs after initial conservative management.Patients whose variceal bleeding is not controlled by 2 injection treatments require more majoremergency surgery."} +{"text": "Drosophila muscle cell fusion takes place both during the formation of the somatic mesoderm and the visceral mesoderm, giving rise to the skeletal muscles and the gut musculature respectively. The core process of myoblast fusion is believed to be similar for both organs. The actin cytoskeleton regulator Verprolin acts by binding to WASP, which in turn binds to the Arp2/3 complex and thus activates actin polymerization. While Verprolin has been shown to be important for somatic muscle cell fusion, the function of this protein in visceral muscle fusion has not been determined.In Verprolin is specifically expressed in the fusion competent myoblasts of the visceral mesoderm, suggesting a role in visceral mesoderm fusion. We here describe a novel Verprolin mutant allele which displays subtle visceral mesoderm fusion defects in the form of mislocalization of the immunoglobulin superfamily molecule Duf/Kirre, which is required on the myoblast cell surface to facilitate attachment between cells that are about to fuse, indicating a function for Verprolin in visceral mesoderm fusion. We further show that Verprolin mutant cells are capable of both migrating and fusing and that the WASP-binding domain of Verprolin is required for rescue of the Verprolin mutant phenotype.Verprolin mutant, however it is not absolutely required for myoblast fusion in either the visceral or the somatic mesoderm.Verprolin is expressed in the visceral mesoderm and plays a role in visceral muscle fusion as shown by mislocalization of Duf/Kirre in the Drosophila muscle progenitors, i.e. myoblasts, arise during embryogenesis and undergo the central process of myoblast fusion during the development of both the visceral and the somatic muscles. The mechanisms underlying cell fusion are actively studied in musculature of Drosophila melanogaster, with significant focus on the process of fusion within the somatic mesoderm (SM), although the phenomenon of myoblast fusion also occurs during the formation of the visceral muscle. The visceral mesoderm (VM) of the fruitfly consists of an inner layer of circular muscles, formed after one round of myoblast fusion, surrounded by an outer layer of longitudinal muscles ,1918,19]5 mutant , indicatTaken together, we suggest that VM fusion is initiated in mutants of components in the Scar-Wasp signaling network, and that these molecules are involved in an increased efficiency of the fusion process.Drosophila, were they have been associated with cellular processes such as vesicle transport and actin-microtubule interactions [In addition to the Arp2/3 complex, other molecular pathways are able to nucleate actin. These include proteins such as formins, Spire and Cordon-bleu. Molecules of these protein families are structurally different to the Arp2/3 complex and produce linear instead of branched actin filaments. , Df(2R)ED3943 , P(Tub-PBac\\T)2/wgSp-1 , rp298lacZ [Vrp1f06715 2/wgSp-1 flies were crossed to Vrp1f06715 flies to induce expression of piggyBac transposase, in order to remobilize the WHf06715 element. To drive LacZ expression in the longitudinal muscles of Vrp1f06715 mutant as well as heterozygous controls, flies with the genotype Vrp1-UAS:lacZ/CyOWgLacZ were crossed to flies with the genotype Vrp1/CyOWgLacZ; 5053-GAL4. For studies of migration and fusion of VM cells in the SM, fly strains with the genotype Alk10-Vrp1/CyOWgLacZ were generated as well as flies with the genotype rp298lacZ;Alk10-Vrp1/CyOWgLacZ . For rescue experiments flies of the genotype Vrp1f06715/CyOWgLacZ;UAS-Vrp1 transgene was used as a PCR template to generate four different myc tagged Vrp1 transgenic constructs; Vrp1 full length (2250 bp), Vrp1 2X\u0394WH2 (1830 bp), Vrp1 \u0394Pro\u0394WBD (450 bp) and Vrp1 \u0394WBD (2140 bp). The primers added a BamHI restriction site to the 5' end of the PCR product and a XhoI restriction site and a myc sequence to the 3' end. Primers for Vrp1 full length were; 5' primer: GGA TCC GCC ATG GCT ATT CCG CCA CCC CCG GGA, 3' primer: CTC GAG CTA CAG ATC CTC TTC AGA GAT GAG TTT CTG CTC CAT ACC ATT GGT GGC CTT AAA. Primers for Vrp1 \u0394WH2 were; 5' primer: GGA TCC GCC GCC ATG ACA ACG AAC TCA TCC GCT CAG, 3' primer: CTC GAG CTA CAG ATC CTC TTC AGA GAT GAG TTT CTG CTC CAT ACC ATT GGT GGC CTT AAA. Primers for Vrp1 \u0394Pro\u0394WBD were; 5' primer: GGA TCC GCC ATG GCT ATT CCG CCA CCC CCG GGA, 3' primer: CTC GAG CTA CAG ATC CTC TTC AGA GAT GAG TTT CTG CTC TTG GCG CTT CAA CGT CAA GTG. Primers for Vrp1 \u0394WBD were; 5' primer: GGA TCC GCC ATG GCT ATT CCG CCA CCC CCG GGA, 3' primer: CTC GAG CTA CAG ATC CTC TTC AGA GAT GAG TTT CTG CTC GGT CTC CAA GTC GTT GAC CAG. Standard PCR programs were used to amplify DNA fragments. PCR products were then digested with BamHI and XhoI and subcloned into the pUAST plasmid [The in situ hybridization a digoxigenin-labelled RNA probe was made using cDNA encoding Vrp1 and a DIG RNA labelling kit (Roche). In situ hybridization of whole-mount wild type Drosophila embryos was carried out as described [Unless otherwise stated, embryos were collected, fixed and immunostained as described previously , prior tescribed .Porcine aortic endothelial (PAE ) cells were cultured in Ham's F12 medium, Supplemented with 10% FBS and penicillin/streptomycin at 37\u00b0C in an atmosphere of 5% CO2. For immunstaining experiments, the cells were seeded on coverslips and transiently transfected by Lipofectamine (Invitrogen Life Technologies) employing the protocol provided by the manufacturer. Twenty hours post-transfection, the cells were fixed in 3% paraformaldehyde in phosphate buffered saline (PBS) for 20 minutes at 37\u00b0C and washed with PBS. The cells were thereafter permeabilized in 0.2% Triton X-100 in PBS for 5 minutes, washed again in PBS and incubated in 5% FBS in PBS for 30 minutes at room temperature. To visualize filamentous actin, cells were incubated with tetramethyl rhodamine isothiocyanate (TRITC)-conjugated phalloidin (Sigma) diluted in 5% FBS in PBS for 30 minutes at room temperature. The coverslips were washed in PBS and mounted on object slides by the use of Fluoromount-G (Southern Biotechnology Associates). Cells were photographed by a Hamamatsu ORCA CCD digital camera employing the QED Imaging System software using a Zeiss Axioplan2 microscope. Thick bundles, actin dots and stress fibers were quantified manually in microscope by calculating the percentage of transfected PAE cells displaying these structures or cells displaying extensive loss of stress fibers Wild type embryo, (B) Vrp1f06715, (C) D-WIPD30, (D) Vrp1f06715/D-WIPD30 transheterozygotes.Click here for fileVrp1f06715 mutant embryos with different Vrp1 constructs, and description of mutant phenotypes observed in PAE cells upon expression of the different Vrp1 constructsSupplemental Figure 2; Rescue experiments of . Rescue of the Vrp1f06715 embryonic mutant phenotype performed with different Vrp1 constructs as described in Figure UAS-Vrp1full length and UAS-Vrp1\u03942xWH2 are both able to fully rescue the SM fusion phenotype of the Vrp1f06715 mutant when expressed with the Sns-Gal4 driver, while UAS-Vrp1\u0394WBD and UAS-Vrp1\u0394Pro\u0394WBD are not. A representative embryo from each cross is shown. Unfused cells are indicated by arrows. (A) Vrp1f06715Sns > > Vrp1full length (B) Vrp1f06715Sns > > Vrp1\u03942xWH2. (C) Vrp1f06715;Sns > > UAS-Vrp1\u0394Pro\u0394WBD. (D) Vrp1f06715;Sns > > UAS-Vrp1\u0394WBD. (E) The white arrow indicates normal stress fibers (SF). Non transfected PAE cells contain numerous stress fibers in contrast to cells that ectopically express full length Vrp1. The Vrp1-expressing cells undergo a very characteristic reorganization of the actin filament system; the cells appear almost empty of the bulk filamentous actin, apart from few and thick bundles of actin filaments and a formation of focal points of actin, so called actin dots. Red arrows indicate the presence of thick bundles and actin dots, as well as stress fiber loss (SF loss).Click here for file"} +{"text": "Heat shock protein (HSP) 47 is associated with collagen type I metabolism, both constitutively and after stress-inflicted injury. It has been claimed that, in contrast to hyperthermia (HT), photodynamic therapy (PDT) does not damage collagen, as measured at the level of tissue. We have studied HSP47 expression in normal murine skin fibroblasts (3T6) treated with hyperthermia or photodynamic therapy (PDT) mediated by three different photosensitizers: (1) haematoporphyrin ester (HpE), (2) meta tetra hydroxyphenyl chlorin (mTHPC) and (3) riboflavin (RB). Riboflavin is not an established photosensitizer for PDT and was chosen here because it is known to provoke collagen damage. The applied doses of the treatments were isoeffective in terms of 3T6 clonogenic cell survival. Analysis, at both transcriptional and translational levels, revealed HSP47 elevation after hyperthermia and after PDT with RB. PDT sensitized by HpE and mTHPC did not significantly alter HSP47 expression. These observations are consistent with our hypothesis that this collagen chaperone is up-regulated by laser-mediated modalities known to damage collagen (i.e. HT and RB PDT) but not by more conventional PDT treatments. Additionally, unexpected significant up-regulation of HSP47 was detected after illumination alone (no photosensitizer) of 3T6 cells at 653 nm laser light, but not at 630 nm."} +{"text": "Mastectomy probably represents over-treatment for the majority of women with screen detected ductal carcinoma in situ (DCIS) and breast-conserving surgery is now widely advocated. In this study, biopsy cavity shavings were used to ensure complete excision in 129 women undergoing breast-conserving surgery for screen detected DCIS. A margin was considered clear if DCIS was > 1 mm from any margin of excision and shavings were clear. Patients with involved margins (DCIS at resection margin) underwent re-excision, irrespective of shaving status. After re-excision, 101 women (78%) had clear margins and 28 (22%) close margins (DCIS < or = 1 mm from resection margin). Cavity shavings were histologically clear of DCIS in all cases. Ipsilateral DCIS recurrence occurred in 12 (9.3%) patients. Two recurrences also contained invasive carcinoma. The median time to diagnosis was 14 months and all recurrences occurred at the site of the previous biopsy. Seven recurrences were detected at the first annual mammogram, four at the second and one at the third. Ipsilateral recurrence was related to margin status; only 2 out of 101 (2%) patients with clear margins recurred, compared with 10 out of 28 (36%) patients with close margins. Local recurrence and close margin status both correlated with a high modified Van Nuys prognostic index score. Our results indicate that local relapse represents residual DCIS rather than true recurrence in the majority of cases. Cavity shavings have proved ineffective in ensuring complete excision. We now ensure a minimum 10 mm margin of excision around all screen-detected DCIS lesions."} +{"text": "Laser treatments were repeated in patients with residualdisease or recurrence for a total of 27 LITT sessions. Four of the 8 patients treated with laserthermal chemotherapy remained alive after a median follow-up of 12 months. Of the 12 tumorsites treated, complete responses were located in the oral cavity (3), oropharynx (1),hypopharynx (1), maxillary sinus (1), and median survival for these patients was 9.5 months.This initial experience with cisplatinum-based laser chemotherapy indicates both safety andtherapeutic potential for palliation of advanced head and neck cancer but this must be confirmedby longer follow-up in a larger cohort of patients.In recent years endoscopically controlled laser-induced thermal therapy (LITT) has beenincreasingly accepted as a minimally invasive method for palliation of advanced or recurrenthead and neck or gastrointestinal cancer. Previous studies have shown that adjuvantchemotherapy can potentiate endoscopic laser thermal ablation of obstructing tumors leadingto improved palliation in advanced cancer patients. Eight patients with recurrent head andneck tumors volunteered to enroll as part of an ongoing phase II LITT clinical trial, and alsoelected to be treated with systemic chemotherapy (cisplatin, 80 mg/m"} +{"text": "A case control study of smoking was carried out on 365 Egyptian males with bladder cancer divided into 278 patients (76%) with previous urinary bilharziasis and 87 (24%) without past infestation. The smoking index was significantly higher in both bilharzial and non-bilharzial patients with bladder cancer than their controls. A significant association was found between heavy and moderate cigarette smoking and bladder cancer developing in non-bilharzial subjects. The local habit of \u201cmeassel\u201d smoking did not differ significantly between bladder cancer patients and controls."} +{"text": "Nitric oxide induced apoptosis in a dose-dependent manner which preceded frank cell death (failure to exclude Trypan blue). These data suggest that epithelial cell death may be NO dependent and via apoptosis, in states of gut inflammation.Chronic inflammation is associated with inducible nitric oxide synthase expression in infiltrating and resident cells and an exaggerated release of nitric oxide. NO can induce apoptosis in macrophages and tumour cell lines. We investigated whether NO induced cell death in an epithelial (T84) cell fine via apoptosis. Culture T84 cells were exposed to a bolus of NO (40 or 80 \u03bcM) dissolved in Hank's balanced salt solution (HBSS) supplemented with 10% fetal calf serum (FCS). After incubation for 4 h at 37"} +{"text": "Haemophilus influenzae type b (Hib) vaccine (relative risk (RR) = 0.73; 95% confidence interval (CI) 0.50\u20131.06). Although similar proportions of cases (12%) and controls (11%) received the polysaccharide Hib vaccine , more controls (41%) than cases (35%) received the conjugate Hib vaccine . Although we found no relationship between most infant vaccinations and subsequent risk of childhood ALL, our findings suggest that infants receiving the conjugate Hib vaccine may be at reduced risk of subsequent childhood acute lymphoblastic leukemia. Further studies are needed to confirm this association and, if confirmed, to elucidate the underlying mechanism. \u00a9 1999 Cancer Research CampaignPrevious studies have suggested that infant vaccinations may reduce the risk of subsequent childhood leukaemia. Vaccination histories were compared in 439 children (ages 0\u201314) diagnosed with acute lymphoblastic leukaemia (ALL) in nine Midwestern and Mid-Atlantic states (USA) between 1 January 1989 and 30 June 1993 and 439 controls selected by random-digit dialing and individually matched to cases on age, race and telephone exchange. Among matched pairs, similar proportions of cases and controls had received at least one dose of oral poliovirus (98%), diphtheria\u2013tetanus\u2013pertussis (97%), and measles\u2013mumps\u2013rubella (90%) vaccines. Only 47% of cases and 53% of controls had received any"} +{"text": "In order to improve the therapeutic effectiveness of photodynamic therapy with Photofrin II and laser light for superficial esophageal cancer, we employed an excimer dye laser instead of an argon dyelaser. Eight superficial esophageal cancer lesions (7 cases) were treated. Of these 8 lesions, 6 werecured by initial treatment, while one lesion required another treatment. The final rate of cure was88% (7/8)."} +{"text": "Agrobacterium tumefaciens is known to cause unexpected phenotypes. Mutations of a specific set of homeotic genes can result in alterred floral structure.Genetic transformation mediated by LeTGA1 and SOLly GLB1) induced by nutrient availability in tomato. To further elucidate their function, we sought to knock out the genes using antisense RNAi. When antisense constructs for the two different tomato genes were each transformed into Micro-Tina tomato plants, one primary transformant with similar mutant flower phenotypes was identified from transformation of each construct. Microarray analysis shows that a similar set of genes were up- or downregulated in both mutants. Sequencing of insertion sites indicates that each is inserted into a repetitive region which could impact expression of affected genes but direct alteration of floral homeotic gene sequences was not detected.Previously we identified two genes (This is the first report that dominant flower mutations could be caused by genetic transformation designed to knock out two nutrient stress related genes. For the last two decades, economically important plants have been genetically transformed for longer shelf life, improved nutritional value, enhanced herbicide tolerance, microbial/insect resistance, and tolerance to various severe environmental stresses Unexpected and undesirable phenotypes are frequently encountered as a result of plant transformation Solanum lycopersicum) genes (LeTGA1 and SOLly GLB1) induced by nutrient stress treatments were identified using cDNA arrays Agrobacterium tumefaciens.Previously, two tomato . The mutants have leafy sepals (TM29 (a SEPALLATA homolog) tomato mutants also exhibits ectopic shoot growth from fruit but these mutants have different flower morphology 0) of a selfing plant To understand the function of a tomato leucine-zipper transcription factor y sepals and appay sepals . Petals y sepals resembley sepals . Anothery sepals . All muty sepals . It has LS1 (bands 1 and 2), two bands for LS3 (bands 3 and 4), suggesting that there may be two insertions in LS3 and one in LS1 (LS1) is partially matched by BI208052 (78% identity) which shares a very low similarity to DNA primase (YP_287459). Band 3 partially matches CV967117 with 75% identity which is 29% identical to the heat shock protein 33. Band 4 sequence is 83% identical to an EST (DB711192) which is not similar to any protein in GenBank. These sequences are presented in LS1 and LS3 insertion sites contain some repetitive sequences ] for that array data and PISTILLATA were affected in both mutants. While the PI homolog is upregulated, the AP1 homolog is downregulated in these mutants exhibits strong up-regulation in both and LS3 and it h mutants . In ArabLeTGA1 and SOLly GLB1 are induced at transcription level by nutrient availability LS mutants contain repetitive sequences while insertion sequence in the control mutant, which does not have the dramatic flower phenotype, is not a repetitive sequence based on GenBank search. This implies that repetitive sequences somehow contributed to the phenotype. It has been suggested that repetitive sequences may serve as either initiators or boundaries for heterochromatin domains It seems likely that the transformation-related changes in the genome may be responsible for this intriguing phenotype. However, it can not be a result of transgene expression because they are antisense constructs for two different genes which do not share any similarity (data not shown). Previously, it has been shown that both SOLly GLB1 antisense construct, gene coding region was amplified using flanking primers of LeHbSac (5\u2032-GAG CTC CAC GAG AAT CAT CAA TCA TGA GTA G-3\u2032) and LeHbXma (5\u2032-CCC GGG TAC AAG TAT TTT GAA CTG ATG ATC AAT-3\u2032). The resulting PCR product of 618 bp was cloned onto pGEM TA Easy vector (Promega). Selected clones were minipreped, digested with SacI and XmaI and cloned into SacI and XmaI digested pBI121. For LeTGA1 , the gene fragment was amplified with LeTGASac (5\u2032-GAG CTC ATG AAT TCT TCAA CAT ATA CTC-3\u2032) and LeTGAXma (5\u2032-CCC GGG AGT GAG CTA AGA GCA CGA AGA CGT-3\u2032). The fragment was 1087 bp and was cloned into pBI121 as above behind the 35S promoter. BLAST analysis revealed no similarity between LeTGA1 and SOLly GLB1sequences. Both constructs were transformed into Agrobacterium tumefaciens strain GV3101 for tomato transformation.Antisense constructs were made using the binary vector pBI121 Agrobacterium tumefaciens GV3101 strain containing a gene construct was cultured on the same day for transformation of these explants the next day. The explants were added to 20 mL of Agrobacterium cell and incubated for 15 minutes with periodic shaking. The explants were then returned to their plates upside down, sealed with micropore tape and incubated at room temperature for two days in subdued light. After this, the explants were transferred into regeneration media to allow for regeneration of shoots. As soon as shoots appeared (about 4\u20138 weeks), they were transferred to rooting medium. After the shoots developed adequate roots, they were transplanted into greenhouse. LS1, LS3, and the control mutants (0 mutant plants.Standard protocol mutants were ide5\u2032-CTA ATA CGA CTC ACT ATA GGG CTC GAG CGG CCG CCC GGG CAG GT-3\u2032 (Ad1) and 2-3\u20325\u2032-P-ACC TGC CC-NH (Ad2) 5\u2032-GGA TCC TAA TAC GAC TCA CTA TAG GGC-3\u2032) 5\u2032-CGT TGC GGT TCT GTC AGT TCC-3\u2032; 23]. In the first PCR cycle, primer extension occurred only from the specific PCR primer that binds to the vector sequence in the DNA fragment within the library. Subsequent PCR using nested primers (IP1nest: 5\u2032-GGTTCTGTCAGTTCCAAACG-3\u2032 and AP2: 5\u2032-AAT AGG GCT CGA GCG GC-3\u2032) complementary to the vector and adaptor sequences generated a DNA fragment. Fragment that did not contain a sequence complementary to the specific primer were not amplified. The PCR products were cloned into a TA-cloning vector pGEM-T Easy (Promega) and sequenced using T7 or SP6 primers. Sequencing was performed at the Penn State Nucleic Acid Facility at University Park, PA.The PCR-based genome walk procedure www.affymetrix.com). The purity of the RNA was determined using a spectrophotometer and integrity was confirmed using gel electrophoresis and visualization of ribosomal bands. RNA samples were labeled using a Superscript Plus Indirect cDNA labeling System (Invitrogen). Hybridization was to a Tom1 tomato cDNA microarray which was the only tomato microarray available at the time (http://bti.cornell.edu/CGEP/CGEP.html). Arrays were hybridized and washed according to the procedures outlined by DeRisi (http://derisilab.ucsf.edu/microarray/protocols.html) using a Biosciences Lucidea Slidepro Hybridizer (Amersham). Arrays were scanned at 532 and 635 nm using a Genepix 4000B (Axon Instruments) and gpr files were created using Genespring GX 7.3 software (Agilent Technologies). Expression levels were determined for the average across the arrays in two different replicates. Gpr files were loess normalized using the R Bioconductor package marray Click here for additional data file."} +{"text": "The oxidative modification of low-density lipoprotein (LDL) in the intima of arteries contributes to the initiation and progression of atherosclerotic lesions. We have previously reported that oxidized LDL (oxLDL) interacts with an endogenous plasma protein, \u03b22-glycoprotein I (\u03b22GPI), to form complexes and that the interaction is mediated by oxLDL specific ligands. We have also demonstrated the presence of oxLDL/\u03b22GPI complexes in the blood stream of patients with systemic inflammatory and autoimmune diseases. These findings implicate that oxLDL/\u03b22GPI complexes are possible atherogenic autoantigens. Autoantibodies to oxLDL/\u03b22GPI complexes have been associated with arterial thrombosis. Further, circulating IgG immune complexes containing oxLDL and 2GPI were also detected in patients with systemic lupus erythematosus (SLE) and/or antiphospholipid syndrome (APS). Although many unanswered questions remain about the exact pathogenic mechanism(s) involved, oxLDL/\u03b22GPI complexes may be described as metabolic products relevant in atherogenesis and as significant participants in autoimmune-mediated atherosclerosis."} +{"text": "Several new complexes of platinum with positively charged cellular dyes have been synthesised in an effort to find chemotherapeutic drugs with increased antitumour cytotoxicity. As part of this effort, the direct cytotoxicities of some of these complexes as well as their ability to inhibit bleomycin potentially lethal damage repair (PLDR) was studied in vitro in a squamous cancer cell line of human origin (SCC-25). All of the new agents were more cytotoxic against exponentially growing than against plateau phase cell cultures. Exposure of cells to non-lethal drug concentrations for between 1 and 6 h led to measurable inhibition of bleomycin PLDR in the case of each drug tested. In order of decreasing ability to inhibit bleomycin PLDR, Pt(fast black)2, Pt(thioflavin)2 and Pt(thionin)2 were more effective than CDDP, while Pt(methylene blue)2, Pt(Rh-123)2 and Pt(pyronin Y)2 were less effective. The most directly cytotoxic agents were Pt(thioflavin)2, Pt(pyronin Y)2 and Pt(Rh-123)2 which also proved to be the least selectively toxic drugs towards exponential versus plateau phase cells. These results indicate that several of the new platinum complexes may be effective cytotoxic agents as well as effective inhibitors of DNA repair process following exposure of cells to other DNA interactive modalities."} +{"text": "We established a panel of 17 xenografts from primary human breast carcinomas. We examined which characteristics of the original tumours and the xenografts facilitate growth in animals. Tumours expressing medium or strong immunoreactivity for p53 protein had significantly (P < 0.05) higher incidence (92%) of in vivo tumour take than those showing weak or negative immunoreactivity (9.1%). No such association was observed between either c-erbB-2 or epidermal growth factor receptor (EGFR) expression in the original tumours and their in vivo tumour take. Following subcutaneous (s.c.) transplantation of original breast tumours or established xenografts, 7/17 tumours showed metastatic disease spread to distant sites (mainly lungs). This study suggests that selective growth of highly aggressive tumours occurs during in vivo propagation of malignant tumours, and these tumours will be of particular interest in evaluating various chemotherapeutic agents for breast cancer management."} +{"text": "Using suppression-subtractive hybridization (SSH) 2 cDNA libraries were created, an UP library (202 cDNA fragments) and a DOWN library (153 cDNA fragments). Using reversed Northern hybridization 16 and 30 fragments were truly differentially expressed in the UP and DOWN libraries, respectively. Most prominent in the UP library were the mitochondrial and injury response clusters and in the DOWN library the cytoskeletal, protein synthesis and signalling clusters. These distinct clusters potentially represent an expression profile of the cisplatin-induced cellular injury response. \u00a9 2001 Cancer Research Campaign\u2002\u2002http://www.bjcancer.comThe goal of this study was to identify changes in mRNA levels in tumour cells after a toxic exposure to cisplatin (IC"} +{"text": "The following funding information was omitted: FW acknowledges financial support from the German Ministry for Education and Research (BMBF) via the Bernstein Center for Computational Neuroscience (BCCN) G\u00f6ttingen, Germany, under Grant No. 01GQ0430."} +{"text": "Subaxial cervical spine dislocations are common and often present with neurological deficit. Posterior spinal fusion has been the gold standard in the past. Pain and neck stiffness are often the presenting features and may be due to failure of fixation and extension of fusion mass. Anterior spinal fusion which is relatively atraumatic is thus favored using autogenous grafts and cages with anterior plate fixation. We evaluated fresh frozen fibular allografts and anterior plate fixation for anterior fusion in cervical trauma.n = 36), C4/5 (n = 15), C6/7 (n = 7) and C3/4 (n = 2). There were 38 unifacet dislocations with nine posterior element fractures and 22 were bifacet dislocations. Twenty-two patients had neurological deficit. Co-morbidities included hypertension (n = 6), non-insulin-dependent diabetes mellitus (n = 2) and asthma (n = 1). All patients were initially managed on skull traction. Following reduction further imaging included Computerized Tomography and Magnetic Resonance Imaging. Patients underwent anterior surgery . All patients were immobilized in a Philadelphia collar for eight weeks (range 7-12 weeks). Eight patients were lost to follow-up within a year. Follow-up clinical and radiological examinations were performed six-weekly for three months and subsequently at three-monthly intervals for 12 months. Pain was analyzed using the visual analogue scale (VAS). The mean follow-up was 19 months (range 14-39 months).Sixty consecutive patients with single-level dislocations or fracture dislocations of the subaxial cervical spine were recruited in this prospective study following a motor vehicle accident. There were 38 males and 22 females. The mean age at presentation was 34 years (range 19-67 years). The levels involved were C5/6 and two patients with root involvement recovered. At six months bony trabeculae at the graft-vertebrae interface were noted. There were 12 (20 %) cases of graft collapse and one case of angulation which showed no progression. At six months the VAS was 3 (range 0-6). There was no limitation of neck motion at six months in 47 patients.Fresh frozen fibular allografts are suitable and cost-effective for anterior fusion in cervical trauma. Subaxial cervical spine dislocations are common and represent significant osteoligamentous disruption, instability and neurological deficit.13111Sixty consecutive patients with single-level dislocations or fracture dislocations of the subaxial cervical spine following a motor vehicle collision between 2004 and 2006 were included into the study. There were 38 males and 22 females. The mean age was 34 years (range 19-67 years). The exclusion criteria included polytrauma, head injury and delayed presentation (>48 h).n = 36), C4/5 (n = 15), C6/7 (n = 7) [n = 2). There were 38 unifacet dislocations with nine associated posterior element injuries and 22 were bifacet dislocations. Twenty-two patients had neurological deficit [n = 6), non-insulin-dependent diabetes mellitus (n = 2) and asthma (n = 1). There were 26 smokers and they smoked an average of 10 cigarettes per day for less than five pack years. All patients were initially managed on skull traction. The patients were positioned supine on a double-split mattress, cone calipers were applied and reduction was effected in the radiology department under conscious sedation and monitoring. Reduction was achieved in 58 patients, the remaining two patients reduced spontaneously following intubation pre surgery. Further imaging included computerized tomography to assess posterior element fractures and magnetic resonance imaging was done to assess for intrinsic cord changes and extrinsic cord compression by a ruptured disc.The levels involved were C5/6 ( (n = 7) and C3/4 deficit . Co-morbPatients underwent anterior cervical surgery between one to two weeks (average seven days post injury). The fibular allografts were harvested and prepared by the national tissue bank (Bone SA) according to the American tissue bank standards of harvesting and storage. A segment measuring 3-5 mm (at a mean cost of $5 per segment) was bevelled to accommodate for the cervical lordosis. The marrow was curetted and both cortices were burred to remove soft tissue remnants. The fibular allograft was then inserted in the prepared disc space and an anterior cervical plate applied. The surgery was performed by the authors and senior residents under supervision. The mean duration of surgery was 75 min (range 64-115 min) and the mean blood loss was 50 ml (range 20-100 ml). There were no wound and neurological complications. All patients were immobilized in a Philadelphia collar for eight weeks (range 7-12 weeks). Bony union was analyzed using the Bridwell criteria by an independent radiologist and orthopedic surgeon. Graft collapse was determined by measuring the graft height initially and at six months. Clinical and radiological examinations were performed six-weekly for three months and subsequently at three-monthly intervals for 12 months. Pain was analyzed using the visual analogue scale (VAS). The neurological status was assessed clinically and documented using the Frankel grading.Eight patients were lost to follow-up within a year. In the remaining 52 patients the mean follow-up was 19 months (range 14-39 months).The mean neurological recovery was 1.1 Frankel grades (range 0-3) and two patients with C5 root involvement recovered. The four patients with severe neurological deficit who had no neurological recovery were elderly with co-morbidities.At three months all patients demonstrated early evidence of graft incorporation which was completed by six months . There wAutologous bone has been favored for use in spinal fusion in both the lumbar and cervical spine due to its superior capacity for incorporation and osteoinduction. The incidence of donor site morbidity following iliac crest bone graft harvesting has been cited as ranging from 8-39%.Successful fusion of cancellous allografts and dynamic anterior cervical plating were observed in 96% of patients at 12 months.717et al.,Butterman et al.,et al.,Samartzis et al.,et al.,et al.,Yue Fresh frozen fibular allografts are suitable and cost-effective (mean of $5 per patient) for anterior fusion in cervical trauma. There was no increased surgery time and donor site morbidity which is associated with the harvesting of autogenous grafts."} +{"text": "Objective: This study was designed to determine whether outpatient treatment of pyelonephritis in pregnancy can reduce costs without compromising safety or efficacy.Methods: Pregnant patients with uncomplicated initial episodes of acute pyelonephritis were considered for outpatient management. The outpatient treatment consisted of an initial dose of IV ceftriaxone (2 g), followed by daily outpatient IM ceftriaxone (2 g) until resolution of fever and flank tenderness, followed by a 10-day course of oral antibiotics. The study group was compared with a group requiring inpatient treatment and a historical control group meeting the criteria for outpatient management but having been treated as inpatients in the previous year.Results: Of the 34 treated as outpatients, only 4 (12%) required hospital admission and 1 developed an upper urinary tract recurrence. None of these patients had premature delivery or any other serious complication. The historical control group (N = 29) included 1 upper urinary tract recurrence, no preterm deliveries, and 1 case of acute respiratory disease syndrome. The outpatient group required an average of 3.4 daily outpatient visits compared with 3.9 days of hospitalization for the historical control group. The inpatient group (N = 39) was significantly more likely to require hospitalization >6 days (P = 0.0004), with a trend toward more frequent upper urinary tract recurrences . The cost analysis revealed a 3-fold difference between outpatient and inpatient therapy .Conclusions: The outpatient treatment of selected patients with pyelonephritis in pregnancy as a promising approach to reducing costs warrants further investigation."} +{"text": "The results were compared to data on proliferation obtained by both flow cytometry (labelling index (LI), the potential doubling time (Tpot) n = 55) and immunohistochemistry , together with immunohistochemical p53 expression (n = 68). There were no overall significant differences in the median values of the various parameters analysed for the different sites within the head and neck region, disease stages, grades of tumour differentiation or nodal states. A subgroup analysis showed that oropharyngeal (n = 22) versus oral cavity (n = 35) tumours were more radiosensitive (P = 0.056) and had a higher Ki-67 index (P = 0.001). Node-positive tumours had higher LI (P = 0.021) and a trend towards lower Tpot (P = 0.067) values than node-negative ones. No correlations were seen between SF2 and any of the parameters studied. The long-standing dogma of an increased radiosensitivity of rapidly proliferating cells in contrast to slowly proliferating cells was not confirmed. The study shows that parallel measurements of different biological markers can be obtained for a large number of patients with head and neck cancers. The independence of the various parameters studied suggests that there may be potential for their combined use as prognostic factors for the outcome of radiotherapy. \u00a9 1999 Cancer Research CampaignA study was made of the relationship between measurements of radiosensitivity versus proliferation and p53 status in head and neck cancers. Inherent tumour radiosensitivity was assessed as surviving fraction at 2 Gy (SF2) using a clonogenic soft agar assay ("} +{"text": "Attenuated APC phenotype is characterized by relatively few colonic polyps, early age at onset of colon cancer compared with the general population, and inactivating germline mutations within specific regions of the APC gene. We hypothesized that germline mutations within these APC gene regions, might contribute to early onset or familial CRC susceptibility. To test this notion, we analysed 85 Israeli patients with either early onset (< 50 years at diagnosis) or familial CRC for harbouring mutations within the relevant APC gene regions: exons 1\u20135, exon 9 and a region within exon 15 using denaturing gradient gel electrophoresis (DGGE), and all of exon 15 employing protein truncation test (PTT). No inactivating, disease-associated mutations were detected in any patient. A novel polymorphism in intron 5 was detected in 16 individuals, 8 patients were carriers of the 11307K variant, a mutation prevalent among Jewish individuals with colorectal cancer, and 4 displayed the E1317Q variant. We conclude that in Israeli individuals with early onset or familial CRC, truncating mutations in the APC gene regions associated with attenuated APC phenotype probably contribute little to disease pathogenesis. \u00a9 2001 Cancer Research Campaign"} +{"text": "Pgk2, Aldoart1, and Aldoart2). Their restricted expression profile suggests that retrotransposition may play a significant role in the evolution of sperm glycolytic enzymes.The central metabolic pathway of glycolysis converts glucose to pyruvate, with the net production of 2 ATP and 2 NADH per glucose molecule. Each of the ten reactions in this pathway is typically catalyzed by multiple isozymes encoded by a multigene family. Several isozymes in this pathway are expressed only during spermatogenesis, and gene targeting studies indicate that they are essential for sperm function and male fertility in mouse. At least three of the novel glycolytic isozymes are encoded by retrogenes and/or provided evidence of alternatively spliced exons. We analyzed expression of sequences with ORFs and <99% sequence identity in the coding region and obtained evidence for the expression of an alternative Gpi1 transcript in mouse spermatogenic cells.We conducted a comprehensive genomic analysis of glycolytic enzymes in the human and mouse genomes and identified several intronless copies for all enzymes in the pathway, except Our analysis detected frequent, recent, and lineage-specific retrotransposition of orthologous glycolytic enzymes in the human and mouse genomes. Retrotransposition events are associated with LINE/LTR and genomic integration is random. We found evidence for the alternative splicing of parent genes. Many retroposed sequences have maintained ORFs, suggesting a functional role for these genes. Although glycolysis is highly conserved, this central metabolic pathway is modified extensively during spermatogenesis. There are several glycolytic isozymes with restricted expression in the male germline including spermatogenic glyceraldehyde-3-phosphate dehydrogenase (GAPDHS) ,2, phospGapdhs and Pgk2) demonstrates an essential role of these enzymes in sperm motility and male fertility in mice HK2 [ENSG00000159399] HK3 [ENSG00000160883], GCK [ENSG00000106633], HKDC1 [ENSG00000156510], GPI1 [ENSG00000105220], PFKL [ENSG00000141959], PFKM [ENSG00000152556], PFKP [ENSG00000067057], ALDOA [ENSG00000149925], ALDOB [ENSG00000136872], ALDOC [ENSG00000109107], TPI1 [ENSG00000111669], GAPDH [ENSG00000111640], GAPDHS [ENSG00000105679], PGK1 [ENSG00000102144], PGAM1 [ENSG00000171314], PGAM2 [ENSG00000164708], PGAM5 [ENSG00000176894], ENO1 [ENSG00000074800], ENO2 [ENSG00000111674], ENO3 [ENSG00000108515], DKFZp781N1041 [ENSG00000188316], PKLR [ENSG00000143627], PKM2 [ENSG00000067225].Human: Hk1 [ENSMUSG00000037012], Hk2 [ENSMUSG00000000628], Hk3 [ENSMUSG00000025877], Gck [ENSMUSG00000041798], Hkdc1 [ENSMUSG00000020080], Gpi1 [ENSMUSG00000036427], Pfkl [ENSMUSG00000020277], Pfkm [ENSMUSG00000033065], Pfkp [ENSMUSG00000021196], Aldoa [ENSMUSG00000030695], Aldob [ENSMUSG00000028307], Aldoc [ENSMUSG00000017390], Tpi1 [ENSMUSG00000023456], Gapdh [ENSMUSG00000057666], Gapdhs [ENSMUSG00000061099], Pgam1 [ENSMUSG00000011752], Pgam2 [ENSMUSG00000020475], Pgam5 [ENSMUSG00000029500], Eno1 [ENSMUSG00000063524], Eno2 [ENSMUSG00000004267], Eno3 [ENSMUSG00000060600], 6430537H07Rik [ENSMUSG00000048029], Pklr [ENSMUSG00000041237], Pkm2 [ENSMUSG00000032294].Mouse: We blasted the mRNA sequence for each gene encoding a glycolytic enzyme using Ensembl BlastView in order to identify retroposed sequences in both the human and mouse genomes. We grouped BLAST hits based upon chromosome location and orientation. Multiple hits in close vicinity and with the same orientation were grouped together in a single hit that span the entire genomic sequences between and upstream/downstream of hits. Hits with less 50 base pairs were excluded. By comparing BLAST results between gene family members, we identified the parent gene for each retroposed sequence. For each retroposed sequence, we identified the parent gene by choosing matches with the longest hit and the highest percentage match. Using the BLAST results, we calculated the weighted average of the nucleotide identity of all retroposed sequences matching glycolytic enzymes. Ensembl was used to retrieve the FASTA sequence for each retroposed sequence on the appropriate strand.http://www.ebi.ac.uk/Tools/clustalw2/index.html-dCTP into the same size product from the parent gene and the retroposed sequences of interest: Tpi1/Tpi1-rs1, Pgam1/Pgam1-rs1, and Eno1/Eno1-rs1. The forward primer sequence for Tpi1/Tpi1-rs1 was 5'CCGACACCGAGGTGGTTT3' and the reverse primer sequence was 5'GTTCTGCGCAGCCACAGCAA3'. The forward primer sequence for Pgam1/Pgam1-rs1 was 5'GCAGACCTCACAGAAGATCAG3' and the reverse primer sequence was 5'ACAGATGTGGTCAGTGTGACAT3'. The forward primer sequence for Eno1/Eno1-rs1 was 5'TTGGGAAAGCTGGCTACACT3' and the reverse primer sequence was 5' CCAGTCATCCTGGTCAAAGG 3'. Arrows in Figure To detect expression of human retroposed sequences, primers were designed to amplify and incorporate \u03b1-[Prm1) in RNA samples. We also included a negative control with no reverse transcriptase as a control for genomic DNA contamination. The forward primer sequence for Prm1 was 5'TCACAGGTTGGCTGGCTC3'and the reverse primer sequence was 5'CATTGTTCCTTAGCAGGCTCC3' [32P]-dCTP into amplified products.As a positive control to confirm proper spermatogenesis in human testis samples, we detected expression of protamine 1 . SDS polyacrylamide gel electrophoresis (SDS-PAGE) on 7.5% polyacrylamide gels was used to separate samples with equal protein amounts, followed by electrophoretic transfer to Immobilon-P PVDF (polyvinylidene fluoride) membranes . Equal protein loading was confirmed by Coomassie blue R250 staining . Membranes were destained, rinsed with TBS-T and incubated in blocking buffer (5% nonfat dry milk in TBS-T) overnight at 4\u00b0C. Antibody incubations were performed at room temperature in blocking buffer. Membranes were incubated with a 1:500 dilution of a polyclonal antibody raised against a recombinant human glucose phosphate isomerase protein fragment for 2 hours. Membranes were then incubated for 45 min at room temperature with secondary antibody diluted 1:10,000. Following antibody incubations, membranes were rinsed for 5 minutes with TBS-T. Immunoreactive proteins were detected by enhanced chemiluminescence using the SuperSignal West Pico substrate and HyBlot CL autoradiography film .Lysis buffer was used to extract proteins from tissues or isolated cells. Samples were centrifuged at 16,000 \u00d7 http://galaxy.psu.edu was used to obtain both 1 kb and 10 kb FASTA format sequence flanking retroposed sequences and genes encoding glycolytic enzymes [http://www.repeatmasker.org. We calculated the percent frequency of repetitive elements in each base pair within 1 kb upstream or downstream of retroposed sequences or genes encoding glycolytic enzymes. Chi-square values were calculated using a contingency table comparing mouse and human sequences versus glycolytic enzymes and retroposed sequences for each repetitive element. (G + C) content was calculated using the eMBOSS geecee program http://inn-temp.weizmann.ac.il/cgi-bin/emboss/geecee[Galaxy enzymes . We analss/geecee.http://www.repeatmasker.org was used to generate sequence with the repetitive elements masked (represented by \"n\") [Repeatmasker by \"n\") . We repehttp://www.ncbi.nlm.nih.gov/projects/homology/maps/. Finally, to determine the evolutionary history of genes within each gene family and their rate of divergence we aligned the coding sequence using ClustalW http://www.ebi.ac.uk/Tools/clustalw2/index.html[http://evolution.genetics.washington.edu/phylip/.We dated the repeated elements by comparing their nucleotide divergence from their respective consensus and then compared these values to the nucleotide divergence of the corresponding retroposed glycolytic sequence to the corresponding parent gene. In addition, we determined whether retroposed sequences are located at homologous position of the human and mouse genome by determining the position of the flanking genes in the appropriate species. We then found the position of the homologous genes in the others species using comparative maps ndex.html and consHk: Hexokinase; Gpi1: Glucose phosphate isomerise; Pfk: Phosphofructokinase; Aldo: Aldolase; Tpi1: Triose phosphate isomerise; Gapdh: Glyceraldehyde phosphate dehydrogenase; Pgk: Phosphoglycerate kinase; Pgam: Phosphoglycerate mutase; Eno: Enolase; Pk: Pyruvate kinase; Ldh: Lactate dehydrogenase; LINE: Long interspersed nucleotide element; LTR: Long terminal repeats; SINE: Short interspersed nucleotide element; SSCP: Single strand conformation polymorphism electrophoresis; ATP: Adenosine triphosphate; RT-PCR: Reverse transcription polymerase chain reaction; ORFs: open reading frames.SAV developed and performed experiments, analyzed data, and drafted the manuscript. FPMV and DAO conceived of the study, participated in experimental design, and edited the manuscript. All authors read and approved the final manuscript.Human retroposed sequences matching genes encoding glycolytic enzymes. This table indicates the gene name for each retroposed sequence in the human genome, along with the chromosome position and strand. FL CDS refers to those sequences containing full-length coding sequence (Y) or only untranslated sequence (UTR), regardless of whether the sequences are in frame. In some cases, multiple retroposed sequences from the same parent gene are located at adjacent chromosome positions. For example ENO1-rs2 and ENO1-rs3 are located less than 2 kb apart on chromosome 15.Click here for fileMouse retroposed sequences matching genes encoding glycolytic enzymes. This table indicates the gene name for each retroposed sequence in the mouse genome, along with the chromosome position and strand. FL CDS refers to those sequences containing full-length coding sequence (Y) or only untranslated sequence (UTR), regardless of whether the sequences are in frame. In some cases, multiple retroposed sequences from the same parent gene are located at adjacent chromosome positions. For example: Eno1-rs15 and Eno1-rs23 are located less than 1 kb apart on chromosome 3 m and Pkm2-rs2 and Pkm2-rs3 are located 150 kb apart on chromosome 2.Click here for fileThe position of genes flanking retroposed sequences in the mouse and human genome. The table identifies the genes flanking each of the retroposed sequences derived from genes encoding glycolytic enzymes. For each retroposed sequenced we determined the position of the flanking genes in the appropriate species and determined the position of the homologous genes in the others species using well established comparative maps http://www.ncbi.nlm.nih.gov/projects/homology/maps/.Click here for fileAmino acid alignment of retroposed sequences in the human genome that maintain ORFs. Asterisks (*) denote identical residues. Methionine residues are highlighted in grey boxes, residues marked as \"X\" in a black box denote stop codons, and dashes denote deleted codons.Click here for fileAmino acid alignment of GPI1-related sequences in the mouse genome that maintain ORFs (GPI1-rs1). Asterisks (*) denote identical residues. Methionine residues are highlighted in grey boxes, dashes denote deleted codons and the stop codon is marked as \"X\" in a black box.Click here for fileAmino acid alignments are shown for retroposed sequences containing upstream start codons. (A) Amino acid sequence alignment comparing upstream extensions of human retroposed sequences to their parent glycolytic enzymes. (B) Amino acid sequence alignment of mouse retroposed sequences with upstream start codons and their parent glycolytic enzymes. Asterisks (*) denote identical residues. Methionine residues are highlighted in grey boxes, residues marked as \"X\" in a black box denote stop codons, and dashes indicate deleted codons. Pkm2-Ss sequence is from [Hs (Homo sapiens), Bt (Bos taurus), Ss (Su scrofa), Mm (Mus musculus). is from . Hs (HomClick here for filePercent frequency of repetitive elements flanking retroposed sequences and genes encoding glycolytic enzymes in the (A) human and (B) mouse genomes. Black bars represent the percent frequency of SINE, LINE and LTR elements flanking all intron-containing parent genes that encode glycolytic enzymes. Gray bars denote the percent frequency of SINE, LINE and LTR elements both upstream and downstream of retroposed sequences derived from these parent genes. White bars represent the genome average frequency of these elements, as determined by Waterston et al., 2002 [l., 2002 .Click here for fileRetroposed and non-retroposed genes evolve at similar rates within each gene family. The figure shows the phylogenetic trees for five gene families . h, denotes human genes; m, denote mouses genes. Black lines are used in branches for which we did not find evidence of retrotransposition. Red lines represent branches with evidence of retrotransposition. Numbers denote the average branch length since the primate/rodent split.Click here for file"} +{"text": "Although the presence of axillary node metastases in breast cancer is a key prognostic indicator and may influence treatment decisions, a significant proportion of patients diagnosed as axillary node negative (ANN) using standard histopathological techniques may have occult nodal metastases (OMs). A combination of limited step-sectioning and immunohistochemical staining (with cytokeratin (MNF.116) and MUC1 (BC2) antibodies) was used to detect OM in a retrospective series of 208 ANN patients. OMs were found in 53 patients (25%), and both step-sectioning and immunohistochemical detection significantly improved detection (P < 0.05). Detection using BC2 (25%) was superior to MNF.116 (18%) and haematoxylin and eosin (H&E) (8%). OMs were found in 51 patients using only the first and deepest sectioning levels and BC2 staining. OMs were more frequently found in lobular (38%) than ductal carcinoma (25%), and more frequently in women less than 50 years (41%) than in older women (19%). Univariate overall and disease-free survival analyses showed that the presence, size and number of OM had prognostic significance as did tumour size (disease-free only) and histological and nuclear grade (P > 0.05). Cox multivariate proportional hazard regression analyses showed that the presence and increasing size of OMs were significantly associated with poorer disease-free survival, independently of other prognostic factors (P < 0.05). However there was not a significant independent association of the presence of occult metastases with overall survival (P = 0.11). These findings have important implications with regard to selection of ANN patients for adjuvant therapy."} +{"text": "Tako-tsubo syndrome (TTS) in its typical and atypical forms is being increasingly recognized in the West owing to early systematic coronary angiography in acute coronary syndromes (ACS).To assess the incidence, the clinical characteristics and the outcome of TTS in a single high volume cath lab in Southern Italy over the last 6 years.Among 1674 consecutive patients (pts) referred to our coronary care units in the last 6 years (2001\u20132006) for ACS we selected 6 (0.5%) pts who fulfilled the following 4 criteria: 1) transient left ventricular wall motion abnormalities resulting in ballooning at contrast ventricolographic or echocardiographic evaluation; 2) normal coronary artery on coronary angiography performed 5 \u00b1 9 hours from hospitalization; 3) new electrocardiographic ischemic-like abnormalities (either ST-segment elevation or T-wave inversion) and 4) emotional or physical trigger event.At admission all pts had presumptive diagnosis of ACS and ECG revealed ST elevation in 3 (50%) and T wave inversion with QT elongation in 3 (50%). In the acute phase cardiogenic shock occurred in 2 (33%) and heart failure in 1(16%). Presenting symptoms were chest pain in 6 (100%), dyspnoea in 2 (33%) and lipotimia in 1 (16%). At echocardiographic-ventricolographic assessment, the mechanical dysfunction was apical in all 6 pts (\"classic\" TTS). In all patients wall motion abnormalities completely reversed within 4.5 \u00b1 1.5 days. The region of initial recovery was the anterior and lateral wall in 4 cases and the lateral wall in 2 cases. Ejection fraction was 35 \u00b1 8% in the acute phase and increased progressively at discharge (55 \u00b1 6%) and at 41 \u00b1 20 months follow-up . All patients remained asymptomatic with minimal treatment.Classic TTS is a frequent serendipitous diagnosis after coronary angiography showed \"surprisingly\" normal findings in a clinical setting mimicking an ACS. Despite its long-term good prognosis life threatening complications in the acute phase can occur. Stress induced cardiomyopathy is described as an acute cardiomyopathy characterized by acute, but rapidly reversible, left ventricle systolic dysfunction in the absence of atherosclerotic coronary artery disease which appears to be triggered by intense psychological and physical stress . This syCharacteristic evolutionary changes that occur over 2 to 3 days include resolution of the ST-segment elevation and subsequent development of diffuse and often deep T-wave inversion that involves most leads. New pathological Q waves may occasionally be observed and, furthermore, it has been possible to observe a prolongation of the corrected QT interval , potentiDuring the last 6 years 2001\u20132006), among 1674 consecutive patients (pts) referred to our coronary care units for ACS, 6 patients presenting at admission with acute onset of cardiovascular event have been retrospectively selected. All fulfilledthe following 4 predetermined criteria to define Tako-Tsubo cardiomyopathy [001\u20132006,All patients underwent, at admission, two-dimensional transthoracic echocardiography, 12 lead electrocardiogram, serial measurements of cardiac isoenzymes including creatine kinase, creatine kinase MB fraction, and troponin. Transthoracic echocardiography has again been repeated at discharge. All patients have been followed-up for a median period of 40 months.All patients underwent comprehensive transthoracic echocardiography examination at rest, upon discharge and at a median period of 40 months follow-up. End-systolic, end-diastolic volumes, ejection fraction and wall motion score index were calculated following the recommendations of the American Society of Echocardiography . The lefValues are expressed as mean \u00b1 standard deviation (SD) unless indicated otherwise. Groups were compared by parametric or non-parametric tests . More than 2 groups were compared using the analysis of variance. Post-hoc tests were performed with the help of Newman-Keuls test. A p < 0.05 was considered as statistically significant.Demographic and clinical features of the study patients are reported in Table Emergency coronary angiography showed angiographically normal coronary arteries in all with ventriculographic confirmation of apical ballooning observed by echocardiogram Figure . Due to We described 6 cases of tako-tsubo cardiomyopathy in a region of Southern Italy close to Naples. In the contemporary definition and classification of the Cardiomyopathies proposed in 2006 by the American Heart Association, stress (\"Tako-Tsubo\") cardiomyopathy is classified among acquired forms of cardiomyopathy . Our datClinicians should consider this syndrome in the differential diagnosis of acute coronary syndromes, especially in post-menopausal women with a recent history of acute emotional or physical stress. The non-invasive suspicion rests on the typical echocardiographic appearance apical ballooning \u2013 usually reversible within a few days or weeks. The angiographic documentation of normal coronary arteries is necessary to substantiate the diagnosis.ABS: Apical ballooning syndromeACS: Acute coronary syndromePt: patientTTS: Tako-tsubo syndrome"} +{"text": "Five injections of urethane, 1 mg./g. body weight to suckling mice markedly reduced the primary immune response against sheep red blood cells assessed by splenic plaque forming cells (PFC) determination and haemagglutinin (HA) titration. The immunological impairment lasted for about 50 days after the end of the treatment. The secondary response tested by HA titration was not affected. A lower dose of urethane (0.5 mg./g.) produced only a delay of the primary HA response. A single neonatal dose of N-nitrosomethylurea (NMU) caused a profound immunodepression evaluated as HA titre and number of PFC. Both primary and secondary responses were still depressed when tested at 50 and 90 days of age respectively. No clear correlation between the degree of immunodepression and lymphoma development was found."} +{"text": "The DNA content of paraffin embedded tumour specimens from 100 children with kidney tumours was studied by flow cytometry. Data of adequate quality were obtained from 93 cases comprising 67 Wilms' tumours with a favourable histology (FH), 12 Wilms' tumours with unfavourable histology (UH) (pleomorphic), 8 bone-metastasising renal tumours of childhood (BMRTC) and 6 rhabdoid renal tumours. Only 4.5% FH compared with 75% UH Wilms' were aneuploid (P less than 0.001). Although BMRTC and rhabdoid tumours are associated with poor prognosis, there were no examples of aneuploidy in these tumours. The proliferation index was found to be of no prognostic value. Staging and ploidy were not correlated with each other in any of the various histological types of renal tumours studied."} +{"text": "Ten benign breast tumours from 9 female patients (8 with fibrocystic disease and 1 with fibroadenoma) and 1 male patient (with gynaecomastia) were processed into slices and individually cultured for 2 days in serum-free Medium 199. [3H]-TdR was added to the culture medium to assess DNA synthesis. The addition of human prolactin to the culture medium (500 ng/ml) significantly (0.05 greater than P greater than 0.01) increased DNA synthesis; all 9 biopsy specimens from the 9 female patients responded positively to this hormone. Ovine prolactin (500 ng/ml) and bovine prolactin (500 ng/ml) increased the mean incorporation of [3H]-TdR into extracted DNA and increased the mean number of [3H]-TdR-labelled cells, but this increase did not reach the 5% level of probability. The sole case of male breast dysplasia analysed in this study did not respond to either human, ovine or bovine prolactin. These results provide evidence that human prolactin and, to a lesser degree, ovine and bovine prolactin are direct mitogenic stimulants to the epithelium in human benign breast tumours."} +{"text": "Telomeres protect the chromosome ends and consist of guanine-rich repeats coated by specialized proteins. Critically short telomeres are associated with disease, aging and cancer. Defects in telomere replication can lead to telomere loss, which can be prevented by telomerase-mediated telomere elongation or activities of the Werner syndrome helicase/exonuclease protein (WRN). Both telomerase and WRN attenuate cytotoxicity induced by the environmental carcinogen hexavalent chromium (Cr(VI)), which promotes replication stress and DNA polymerase arrest. However, it is not known whether Cr(VI)-induced replication stress impacts telomere integrity. Here we report that Cr(VI) exposure of human fibroblasts induced telomeric damage as indicated by phosphorylated H2AX (\u03b3H2AX) at telomeric foci. The induced \u03b3H2AX foci occurred in S-phase cells, which is indicative of replication fork stalling or collapse. Telomere fluorescence in situ hybridization (FISH) of metaphase chromosomes revealed that Cr(VI) exposure induced an increase in telomere loss and sister chromatid fusions that were rescued by telomerase activity. Human cells depleted for WRN protein exhibited a delayed reduction in telomeric and non-telomeric damage, indicated by \u03b3H2AX foci, during recovery from Cr(VI) exposure, consistent with WRN roles in repairing damaged replication forks. Telomere FISH of chromosome spreads revealed that WRN protects against Cr(VI)-induced telomere loss and downstream chromosome fusions, but does not prevent chromosome fusions that retain telomere sequence at the fusion point. Our studies indicate that environmentally induced replication stress leads to telomere loss and aberrations that are suppressed by telomerase-mediated telomere elongation or WRN functions in replication fork restoration. Telomeres are highly specialized chromatin structures consisting of tandem repeats of the TTAGGG sequence bound and regulated by telomeric proteins (shelterin) and a plethora of accessory factors. Located at the ends of linear chromosomes, telomeres prevent the DNA damage response (DDR) and repair machineries from recognizing and processing the ends as double-strand breaks (DSBs) Accumulating evidence indicates that replication fork stalling or collapse at telomeric ends can lead to telomere loss or aberrations. Telomeric instability associated with defects in telomere replication are induced by polymerase inhibitors and agents that stabilize DNA G-quadruplexes or by depletion of shelterin TRF2 or POT1 proteins in vitro induces DNA polymerase arrest with the most potent arresting lesions at G runs The environmental metal hexavalent chromium (Cr(VI)) is an important source of DNA replication stress. The inhalation of Cr(VI) particles is strongly linked to respiratory cancers in the occupational setting In this study we examined whether Cr(VI)- induced replication stress impacts telomere integrity at the molecular and chromosomal level. We further tested the hypothesis that WRN and telomerase protect against Cr(VI)-induced cytotoxicity and genomic instability, partly by preventing Cr(VI)-induced telomere defects. We found that Cr(VI) exposure in human cells induces telomeric damage as indicated by telomere dysfunction induced foci (TIFs), and telomeric abnormalities associated with defects in telomere replication including telomere loss on chromatids. The latter was attenuated by telomerase expression, and exacerbated by WRN depletion. Thus, we provide novel evidence that environmentally induced replicative stress can impact telomere integrity, and offer a mechanistic explanation for the increased sensitivity of WRN and telomerase deficient cells.Previous studies showed that exposure to Cr(VI) significantly reduced survival of TERT- BJ cells compared to the TERT+ BJ cells in situ hybridization (Telo-FISH) with a PNA probe (To determine whether Cr(VI) exposure can induce telomeric defects and whether telomerase can protect against such telomere instability, we examined individual telomeres in TERT- and TERT+ BJ cells after 0.5 \u00b5M Cr(VI) exposure for 48 h, followed by 10 h recovery. Telomeres on metaphase chromosome spreads were visualized by telomere fluorescent NA probe [31]. TeNA probe . ImportaNA probe . AdditioNA probe . In agreNA probe , but notOccupational Cr(VI) exposure poses a well established risk for developing lung cancer Telomere dysfunction-induced foci (TIFs) refer to the colocalization of DNA damage response factors with telomeres, and are widely used to investigate factors that induce telomere instability including compromised shelterin proteins or G4 stabilizing agents Low levels of Cr(VI) exposure for 6\u201324 h induces replicative stress, indicated by \u03b3H2AX association with S phase cells To directly evaluate the consequence of Cr(VI)-induced telomere damage on telomere structure and integrity we stained individual telomeres on chromosome metaphase spreads using the Telo-FISH assay . The totBiochemical and cellular evidence support WRN roles in facilitating telomere replication, and in recovery from replication fork stalling Upon Cr(VI) exposure WRN re-localizes from the nucleoli into nucleoplasmic foci in S-phase cells that co-localized with \u03b3H2AX foci Next, we asked whether the pattern of WRN foci localization is associated with Cr(VI)-induced telomere damage, as indicated by TIFs. The Cr(VI) cellular exposure experiment in To test whether WRN functions in repair of Cr(VI)-induced telomere damage, we employed U2OS cell lines that stably express either a short hairpin (sh) RNA targeting WRN mRNA or a control shRNA Based on the evidence that WRN depleted cells exhibit reduced recovery from telomere damage -5, we asTo directly test Cr(VI)-induced telomeric defects in WRN depleted cells, compared to WRN proficient cells, the shWRN and shCtrl U2OS cell lines were exposed to 3 \u00b5M Cr(VI) for 48 h and recovered in Cr(VI)-free media with colcemid for 10 h. We observed several types of telomeric defects Recent reports indicate that WRN prevents the induction of chromatid breaks at fragile sites due to replication fork stalling Telomere instability is linked to human diseases and cancer development, and several reports indicate that telomeres are vulnerable to oxidative and replication stress. Cr(VI) is an environmental lung carcinogen that induces lesions that interfere with DNA replication. In this study, we show that Cr(VI) exposure leads to telomere damage and chromosomal telomere loss and aberrations associated with Cr(VI)-induced replication stress. Telomerase expression alleviates Cr(VI)-induced telomere instability, and may explain the hypersensitive phenotype of telomerase negative cells to Cr(VI) toxicity. WRN protein promotes replication fork recovery and telomere replication Telomerase confers cellular resistance to genotoxins with different modes of action 2 to mimic physiological conditions and minimize oxidative stress induced by culture conditions, and while oxidative stress causes accelerated shortening of mean telomere lengths Together with previous work our findings support a role for Cr(VI)-induced replication stress in generating telomeric defects, rather than oxidative stress which can be induced by genotoxic metals and can target telomeres for damage. Guanine is particularly susceptible to oxidative damage and chronic oxidative stress accelerates telomere attrition WRN is implicated in telomere preservation by resolving alternate structures at telomeres during replication, and by restoring stalled or broken replication forks WRN prevents stochastic telomere loss presumably by facilitating telomere replication and resolving alternate structures In summary, our data show that environmentally induced replication stress can lead to telomeric aberrations and instability that are attenuated by telomerase expression or WRN protein activity. Furthermore, our data suggest that telomeric aberrations contribute to Cr(VI) induced cytotoxcitiy and genotoxicity, and may contribute to respiratory cancers resulting from Cr(VI) exposure. Thus, we provide novel evidence that an environmental pollutant can induce telomere instability, which may contribute to environmentally relevant diseases including cancer.2 and 5% O2 at 37\u00b0C. Human U2OS osteosarcoma cell line (ATCC) was cultured similarly except with 10% FBS Werner syndrome (WS) skin fibroblasts (AG03141) and WI-38 lung fibroblasts were from the Coriell Institute . The telomerase-immortalized WS cell line (AG03141) was a gift from Dr. Junko Oshima (University of Washington). BJ and telomerase-immortalized BJ (hTERT BJ) skin fibroblasts were kindly provided by Dr. Peter Lansdorp (University of British Columbia). Cells were cultured in Dulbecco's Modified Eagle Media supplemented with 15% fetal bovine serum, penicillin (50 units/ml), and streptomycin (50 \u00b5g/ml) in humidified chambers with 5% CO2Cr2O7; Cr(VI) as described previously Cells were exposed to potassium dichromate depending on Cr(VI) concentrations, were seeded in 6-cm culture dishes and incubated overnight. After Cr(VI) exposure, the cells were cultured in Cr(VI)-free medium for 7 days. Then cells were stained for 15 min, and colonies composed of 25 or more cells were counted. The survival fraction at each Cr(VI) concentration was determined by dividing the average number of colonies on treated plates by the average number of colonies on untreated plates after adjusting for the initial seeding cell number (plating factor). Each Cr(VI) concentration exposure was performed in triplicate for each of four independent experiments.The cell viability assay (CVA) was conducted as previously described with slight modification The association of \u03b3H2AX with S-phase cells was detected by double immunostaining with antibodies against \u03b3H2AX and incorporated BrdU as previously described with slight modification 2 buffer , and 0.5 \u00b5g/ml Cy3-OO-(CCCTAA)3 PNA probe ). After 2 h hybridization at room temperature, the samples were washed twice with wash solution (70% deionized formamide and 10 mM Tris-HCl [pH 7.4]). Samples were counterstained with DAPI, mounted onto slides and images were acquired with an Olympus FluoView 1000 confocal microscope as described previously The IF-FISH assay was performed as described previously with modification 5), BJ (2.0\u00d7105), hTERT BJ (6.0\u00d7104), shWRN U2OS (1.0\u00d7105) or shCtrl U2OS (1.0\u00d7105) cells were seeded in 10-cm culture dishes and incubated for 2 days. After Cr(VI) exposures, the cells were treated with 0.05 \u00b5g/ml colcemid (Invitrogen) for 10 h. Telomere FISH on metaphase chromosomes was performed as described previously with some modification WI-38 equipped with PlanApo 60\u00d7/1.40 oil immersion objective. The NIS element advanced software was used to acquire and analyze the images with the same settings for paired cell lines in each experiment. In order to rigorously identify and qualify telomere staining and telomere signal free chromosome ends, fusions and aberrations, a series of z-stacked images (0.15 \u00b5m steps) were acquired for each metaphase and analyzed. This technique allowed for rigorous distinction of a telomere signal that was lost from a telomere signal that was out of focus.All statistical analyses were conducted with SAS software . Student t-test was used to determine the significance of differences between two treatments or time points. To determine the significance of differences among more than two treatments or time points, one-way ANOVA followed by Duncan's multiple comparison test was employed. The statistically significant level was set at p<0.05.Table S1Percent telomere defects (number of defects/number of chromosomes). The cells were exposed to the indicated Cr(VI) doses for 48 h and cultured in Cr(VI)-free medium for 10 h.(0.70 MB EPS)Click here for additional data file.Figure S1WRN does not prevent TIF formation. Confocal images of shCtrl and shWRN U2OS cells exposed to 4 \u03bcM Cr(VI) for 48 h (A) and then cultured in Cr(VI)-free medium for 12 h (B). (C) Average \u03b3H2AX foci and TIF number per cell and the percent of TIF positive cells from (A). The data represent mean \u00b1SE from two independent experiments, based on at least 50 randomly chosen cells for each Cr(VI) treatment.(1.96 MB EPS)Click here for additional data file.Figure S2WRN deficiency does not affect Cr(VI)-induced doublets and telomeric DNA-containing double minute chromosomes (TDMs). shCtrl and shWRN cells were exposed to 0 and 3 \u03bcM Cr(VI) for 48 h and cultured in Cr(VI)-free medium for 10 h. (A) Average doublets per chromosome. (B) Average TDMs per chromosome. Around 40 metaphases from two independent experiments were analyzed to quantitate Cr(VI)-induced telomere instability.(0.70 MB EPS)Click here for additional data file."} +{"text": "Human chromosome 8q24 has been implicated in prostate tumorigenesis.Consequently, we evaluated seven 8q24 sequence variants relative to prostate cancer (PCA) in a case-control study involving men of African descent. Genetic alterations were detected in germ-line DNA from 195 incident PCA cases and 531 controls using TaqMan polymerase chain reaction (PCR).Inheritance of the 8q24 rs16901979 T allele corresponded to a 2.5-fold increase in the risk of developing PCA for our test group. These findings were validated using multifactor dimensionality reduction (MDR) and permutation testing (p = 0.038). The remaining 8q24 targets were not significantly related to PCA outcomes.Although compelling evidence suggests that the 8q24 rs16901979 locus may serve as an effective PCA predictor, our findings require additional evaluation in larger studies. Prostate cancer (PCA) accounts for 25% of all diagnosed cancer cases and was the second leading cause of cancer-related deaths in 2009 among U.S. men . Well-esSeveral genetic variants or single nucleotide polymorphisms (SNPs) in the 8q24 region have been associated with PCA risk in Caucasians; however, similar studies on African Americans are only represented in a handful of published reports -8. AmundUnfortunately, the impact of these two loci and other 8q24 markers on PCA risk among men of African descent remains largely unknown. Although the 8q24 rs10090154T allele confers a 1.7-fold increase in PCA risk in a small case-control study set involving African Americans (85 cases and 149 disease-free men), this locus only reached borderline significance .To clarify the role of 8q24 sequence variants in PCA susceptibility among men of African descent, we sought to confirm previous reports and generate new data on the role of seven 8q24 sequence variants in PCA risk among 864 men of African descent (195 cases and 531 disease-free men). To overcome sample size issues and control for multiple comparisons, we used a multifactor dimensionality reduction (MDR) algorithm along with permutation testing. This approach will aid future studies that analyze different sequence variants within the 8q24 region to decode health disparities in high-risk subgroups, especially PCA risk in men of African descent.Between 2001 and 2005, 864 unrelated male residents were recruited from the Washington, D.C. and Columbia, SC areas through the Howard University Hospital (HUH) Division of Urology or PCA screening programs. The study population of men of African descent consisted of 195 incident PCA cases and 531 controls. PCA patients between the ages of 41 and 91 were diagnosed within one year of enrollment. Following a visit to the HUH Division of Urology for an annual PCA screening exam or urinary symptoms, incident PCA cases were identified by a urologist using a transrectal ultrasound-guided biopsy . Biopsy (1) rs6983561 (A > C); (2) rs1447295 (G > T); (3) rs4242384 (A > C); (4) rs4242382 (G > A); (5) rs11934905 (G > A); (6) rs16901979 (G > T); and (7) rs10090154 (G > A). Each allelic discrimination assay contained approximately 40 ng of germ-line DNA, 1\u00d7 Universal Master Mix , a 40\u00d7 mixture containing 900 nM of each primer (forward and reverse), and 200 nM of each probe (FAM and VIC) to comprise a 5 \u03bcl reaction. To facilitate amplification of regions containing the aforementioned 8q24 SNPs, primers and probes were designed in our laboratory using sequences provided by NCBI [Polymorphisms detected in the 8q24 region were ascertained using TaqMan Polymerase Chain Reaction (PCR) allelic discrimination assays. The following seven alleles were detected: One hundred ancestry autosomal markers were included to account for potential population stratification among our admixed population of self-reported African-Americans, West African Americans, East African Americans, and Afro-Caribbean Americans, as previously described . Study p(1) rs6983561 (A/A); (2) rs1447295 (G/G); (3) rs4242384 (A/A); (4) rs4242382 (G/G); (5) rs11934905 (G/G); (6) rs16901979 (G/G); and (7) rs10090154 (G/G). For rs11934905, hetero- and homozygotes (AA + GA) were combined due to the low frequency of the minor allele. Test for trend included genotypes as ordinal variables. Statistical significance was assessed using a P-value < 0.05. All chi-square tests and LR analyses were conducted using the SAS 9.1.3 software .To assess whether inheritance of at least one variant 8q24 allele was associated with an elevated risk of developing PCA, we tested for significant differences in the distribution of seven 8q24 genotypes between 195 cases and 531 controls using the chi-square test of homogeneity. Associations between PCA risk and 8q24 sequence variants, expressed as odds ratios (ORs) and corresponding 95% confidence intervals (CIs), were estimated using unconditional multivariate LR models adjusted for potential confounders . All reported risk estimates and 95% CIs for the selected 8q24 loci used the following as reference genotypes: ths of the data, the average CVC from the observed data was compared to the distribution of average consistencies under the null hypothesis of no association. Validation of models as effective predictors of PCA susceptibility was derived empirically from 10,000 permutations. This approach accounted for multiple testing issues as long as the entire model fitting procedure was repeated for each randomized dataset to provide an opportunity to identify false-positives. We considered MDR permutation results to be statistically significant at the 0.05 level.Multifactor dimensionality reduction (MDR) was used to evaluate and validate main effects associated with PCA risk. This algorithmic tool aids in the identification of high-risk markers using a cross validation strategy to estimate the classification and prediction accuracy of individual factor models ,17. MDR The patient and tumor characteristics in the current study are summarized in Table Our laboratory successfully completed the analysis of seven 8q24 sequence variants among 864 men of African descent (195 PCA cases and 531 controls). Inheritance of at least one minor 8q24 minor allele was fairly common among controls with frequencies ranging between 4.0 and 66.3%, as detailed in Table The independent effects of genetic variations detected within the 8q24 region were analyzed relative to PCA susceptibility using MDR and unconditional LR multivariate models adjusted for age and West African ancestry and advanced PCA among carriers of 8q24 rs4242382A and rs4242384C alleles [Several of the evaluated 8q24 SNPs may be more appropriate as PCA predictors among men of European- rather than African descent ,6,11,23. alleles . To our We have considered the strengths and limitations of the current study. MDR controls for multiple comparisons and spurious risk estimates by using a cross validation and permutation testing scheme as a built-in feature. Misclassification by case status is also a potential limitation for this study. There is a possibility that some men who presented as disease-free following the initial diagnostic evaluation may eventually develop PCA. In an attempt to ease case-status misclassification, controls with at least one abnormal diagnostic test underwent multiple core needle biopsies. Those with an abnormal biopsy were reclassified as cases. Men who received a normal biopsy test but had an abnormal PSA (> 4.0 ng/ml) and/or irregular DRE (n = 48) were excluded because we could not predict with any level of certainty whether they would eventually develop PCA. For similar reasons, we also excluded individuals (n = 65) who were diagnosed with BPH following biopsy. Notably, after close inspection of PCA tissue, it is feasible to overlook a microscopic nodule that can later develop into cancer ; howeverAnother challenge for genetic epidemiology studies involving study participants of African descent is their unique population history of gene flow from divergent populations ,28. The Finally, it is feasible that the observed association is related to linkage disequilibrium of rs16901979 with unknown targets in this region. To address this issue, future studies will use next generation sequencing techniques to better consider the complete heterogeneity within this locus relative to PCA outcomes. Emphasis will be placed on sequence variants within region 3, as it is speculated that sequence variants may be related to a 2-fold increase in PCA risk ,13,29. OIn summary, the 8q24 rs16901979 locus has a strong genetic linkage to PCA among men of African descent in the current investigation as well as other studies. Future multicenter collaborative efforts will facilitate the identification and validation of a reliable panel of genetic susceptibility biomarkers with the capacity to improve PCA detection and prognosis strategies, ultimately reducing PCA health disparities among all men.The authors declare that they have no competing interests.Taqman PCR reactions were carried out in an ABI Prism 7900HT Sequence Detection System (Applied Biosystems) by MLB, TTV, and NAL. TTV designed the primer and probe sequences for Taqman allelic discrimination assays. MLB and LRK performed statistical analysis and produced drafts of this manuscript. All authors made contributions toward editing the manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2407/10/334/prepub"} +{"text": "A murine ascites tumour was treated with intraperitoneal haematoporphyrin derivative (HPD) and laser light . HPD was given intraperitoneally 2 hours before 16 minute laser treatment. Uptake studies 2 hours after HPD injection showed 5-12 fold greater concentration of HPD in tumour cells than in 4 different normal tissues. A total of four HPD/laser treatments, given at 2 day intervals, resulted in 100% complete response; the cure rate was 85%. This study illustrates the effective use of intraperitoneal photodynamic therapy and opens the possibility of exploring different sensitizers, excitation wavelengths, and delivery systems in the treatment of human ascites tumours."} +{"text": "Breast cancer prognosis has previously been linked to the degree of tumour vascularisation. In order to establish additional markers for tumour angiogenesis, we have used monoclonal antibodies against the endothelial Tie receptor tyrosine kinase to study the degree of vascularisation of breast carcinomas and the regulation of Tie expression in the vascular endothelial cells. Antibodies were used for Tie detection and the results were correlated with other prognostic markers. Of four monoclonal antibodies directed against different epitopes of the Tie extracellular domain, two reacted against Tie in unfixed histopathological sections of breast carcinomas. One of these antibodies (clone 7e8) was specific for the endothelial cells whereas the other (clone 10f11) also reacted with basement membranes and occasional carcinoma cells. When Tie expression was studied with the antibody clone 7e8, all 27 carcinomas, two in situ carcinomas, samples of histologically normal breast tissue (n = 16) or normal skin or lymph node tissue (n = 5) showed staining. Microvessel counts were higher in carcinomas than in fibrodenomas or histologically normal breast tissue . A similar result was obtained using antibodies against the CD31 (PECAM) antigen. Microvessel counts in 7e8 staining were not significantly associated with primary tumour size, axillary nodal status, histological grade or staining for oestrogen receptor, progesterone receptor, Ki-67 proliferation marker or p53 oncoprotein."} +{"text": "Vascular endothelial growth factor (VEGF) is known to be produced by various solid tumours and is thought to be involved in microvascular permeability and/or angiogenesis. To examine the relationship between VEGF expression in ovarian neoplasms and clinicopathological factors or patient survival, expression of VEGF was analysed immunohistochemically in 110 epithelial ovarian tumours. In addition, VEGF levels in the tumour fluid (17 patients), ascites (12 patients) and sera (38 patients) were determined using enzyme immunoassay. Positive immunostaining for VEGF was observed in 97% (68 out of 70) of ovarian carcinomas, which was significantly higher than that of tumours of low malignant potential (LMP) and benign cystadenomas (P < 0.01). In ovarian carcinomas, strong VEGF immunostaining was also observed more frequently in tumours of clear cell type (P < 0.05) in the advanced stage of disease (P < 0.05) and with positive peritoneal cytology (P < 0.01). Patients with strong VEGF staining had poorer survival rates than those with weak or no immunostaining for VEGF (P < 0.01). These findings suggest that strong VEGF expression plays an important role in the tumour progression of ovarian carcinoma. The enzyme immunoassay revealed higher serum VEGF levels in carcinoma patients than those in patients with LMP or benign tumours (P < 0.01). Serum VEGF levels decreased after the successful removal of tumours in ovarian cancer patients and, in one patient, the serum VEGF level was re-elevated during relapse. Therefore, serum VEGF could be used as a marker for monitoring the clinical course of ovarian cancer patients."} +{"text": "DNA measurements on biopsy material from 24 squamous cell carcinomas of the oral cavity given preoperative radiotherapy indicate that DNA aneuploid tumours respond better to radiotherapy than do diploid and polyploid tumours. The mean S-phase value was higher (16.1%) for 8 tumours that were eradicated by preoperative radiotherapy than for 13 that did not respond (8.1%). These factors correlated better with the response than did histological and clinical (T) classifications. DNA-ploidy and S-phase estimation can complement the histological diagnosis, and may prove valuable when planning treatment."} +{"text": "Aberrant Wnt gene expression is involved in the development of breast cancer, but its role in other tumours is unknown. Wnts regulate cadherin function, previously shown to be more commonly deregulated in invasive bladder cancer. This study investigated whether factors upstream of cadherins were aberrantly expressed in superficial bladder cancer. The expression of one transforming (Wnt7b) and one non-transforming (Wnt5a) Wnt gene in four human bladder carcinoma cell lines, and in normal human bladder tissues (n = 8) and bladder cancers (n = 48) were analysed by ribonuclease protection analysis. All cell lines expressed an approximately equal level of Wnt7b mRNA. Wnt5a and Wnt7b mRNAs were both expressed in normal bladder tissues and bladder tumours. The median expression of Wnt7b was fourfold higher in superficial tumours (n = 29) than in normal tissues and five fold higher than in invasive tumours . There was no significant difference between normal tissues and invasive tumours (P = 0.3). The expression of Wnt5a did not vary significantly between normal tissues and superficial tumours (P = 0.4), normal tissues and invasive tumours (P = 0.3) or superficial tumours and invasive tumours (P = 0.2). The differential expression of Wnt7b suggests a role in the early events of superficial bladder tumorigenesis involving cell adhesion and provides further evidence of different pathways of evolution of superficial and invasive cancer."} +{"text": "We studied peripheral blood mononuclear cells from 50 patients with active B- and T-cell non-Hodgkin's lymphoma by DNA hybridisation. Nineteen patients (38%) had circulating clones of cells detected by immunoglobulin gene rearrangement (17 patients) or T-cell receptor gene rearrangement (2 patients) with JH and J beta 2 probes. Lymphoma tissue and peripheral blood were studied simultaneously in 22 patients, 9 of which had a circulating clone of cells in peripheral blood. In 7 patients the gene rearrangement in lymphoma tissue and peripheral blood mononuclear cells was identical. However, in 2 patients both heavy chain and light chain gene rearrangements were different in tissue and peripheral blood. The incidence of peripheral blood involvement was commonest in advanced CSIII & IV disease (54%) compared to CSI & II disease (18%) (P less than 0.05), and in low grade (45%) compared to intermediate and high grade lymphoma (31%) . Only 4 patients had definite lymphoma cells seen on peripheral blood smear. The presence of circulating lymphoma cells correlated with conventional assessment of bone marrow involvement although circulating clones were detected in 30% (12/40) of patients with apparently normal bone marrow."} +{"text": "X death time, (2) protamine was more toxic for leukemic than normal mice and (3) the in vitro reaction between Rauscher virus and protamine reduced its infectivity for mice.This study indicated that: (1) i.p. inoculation of protamine into (Rauscher) leukemic mice increased their"} +{"text": "The aims of the trial were to establish the response rate and determine the toxicity of combination chemotherapy with ifosphamide, vincristine and cisplatin (HOP regimen) in advanced metastatic seminoma and to study the role of post-chemotherapy consolidation treatment. Patients with bulky metastatic non-alpha-fetoprotein-producing seminomas were eligible for this phase II study [serum human chorionic gonadotropin < 200 U l-1 (< 40 ng l-1)] if they presented with abdominal masses > or = 10 cm or had extra-gonadal seminoma or had relapsed after previous radiotherapy. The HOP regimen consisted of four 3-weekly cycles of the following drug combination: ifosphamide , vincristine and cisplatin . Residual masses persisting 6 months after chemotherapy could be considered for consolidation surgery or radiotherapy. Maximal response to the HOP chemotherapy was based on the WHO criteria. The median observation time was 2.5 years (range 1.8-5.5 years). Thirteen institutions treated 42 eligible patients within the study . Two patients were not evaluable for response owing to premature treatment discontinuation. Maximal response was as follows: complete remission (CR), 26 (65%); partial remission (PR) 11 (28%); no change (NC), 2 (5%); progressive disease (PD), 1 (3%). Four patients have died, three from their malignancy (two without previous irradiation and one with prior radiotherapy). The fourth patient died of treatment-related toxicity. The 3 year survival for all 42 eligible patients was 90%. Dose reduction and treatment postponement were necessary in 25 and 14 patients respectively. Ten patients experienced granulocytic fever. Previously irradiated patients tolerated chemotherapy as well as non-irradiated patients. Immediately after HOP chemotherapy a mass persisted in 16 of 17 patients with retroperitoneal masses of > or = 100 mm at presentation. Three of these residual lesions were resected within the following 6 months showing complete necrosis. Four lesions dissolved spontaneously during the first year of follow-up. Nine lesions persisted for > or = 1 year (one after consolidation radiotherapy) without leading to relapse. Four of seven patients with mediastinal lesions achieved CR and three a PR after HOP chemotherapy. The HOP chemotherapy regimen is highly effective in patients with advanced metastatic seminoma or those relapsing after previous radiotherapy, but is associated with a high risk of toxicity, in particular myelotoxicity."} +{"text": "P< 0.001). The positivity of hTERT for HCC and corresponding non-cancerous liver was 100% and 30.4% respectively (P< 0.001). Seventy-four per cent (17/23) of HCCs showed strong hTERT expression, but none of the non-cancerous liver tissues did. hTERT expression of the 21-gauge needle biopsied specimens showed no significant difference from that of the surgical samples. The present study revealed that hTERT is strongly expressed in most HCCs, and that hTERT but not hTEP1 is a key component regulating telomerase activity in human liver. \u00a9 2000 Cancer Research CampaignTo know whether two protein components of human telomerase (human telomerase-associated protein 1 (hTEP1) and human telomerase reverse transcriptase (hTERT) are useful markers for telomerase activation in human liver diseases, we examined mRNA levels of these and telomerase activity in human liver samples. Twenty-three human hepatocellular carcinomas (HCCs) and corresponding adjacent livers were analysed for hTEP1 and hTERT expression by semiquantitative reverse transcription-polymerase chain reaction, and for telomerase activity by a telomeric repeat amplification protocol assay. Thirteen liver samples (ten HCCs and three dysplastic nodules) that were biopsied with 21-gauge needles were analysed for hTERT expression. hTEP1 was expressed in all samples examined. No correlation between hTEP1 expression and telomerase activity was observed. hTERT expression significantly correlated with telomerase activity ("} +{"text": "The role of the neurotrophin regulated polypeptide, VGF, has been investigated in a rat spared injury model of neuropathic pain. This peptide has been shown to be associated with synaptic strengthening and learning in the hippocampus and while it is known that VGFmRNA is upregulated in dorsal root ganglia following peripheral nerve injury, the role of this VGF peptide in neuropathic pain has yet to be investigated.Prolonged upregulation of VGF mRNA and protein was observed in injured dorsal root ganglion neurons, central terminals and their target dorsal horn neurons. Intrathecal application of TLQP-62, the C-terminal active portion of VGF (5\u201350 nmol) to na\u00efve rats caused a long-lasting mechanical and cold behavioral allodynia. Direct actions of 50 nM TLQP-62 upon dorsal horn neuron excitability was demonstrated in whole cell patch recordings in spinal cord slices and in receptive field analysis in intact, anesthetized rats where significant actions of VGF were upon spontaneous activity and cold evoked responses.VGF expression is therefore highly modulated in nociceptive pathways following peripheral nerve injury and can cause dorsal horn cell excitation and behavioral hypersensitivity in na\u00efve animals. Together the results point to a novel and powerful role for VGF in neuropathic pain. The spontaneous burning pain, hyperalgesia and allodynia that characterize neuropathic pain are triggered and maintained by a combination of peripheral and central processes ,2. PeripAt the heart of many of these processes lie the neurotrophins, which in addition to controlling the survival and differentiation of neurons, play a key role in maintaining and modulating the function of adult nociceptive neurons. NGF and BDNF are highly regulated in skin, peripheral and central neurons, and glia following nerve injury and tissue inflammation, and have been repeatedly implicated in the development and maintenance of chronic neuropathic pain states -10.vgf gene transcription in vitro and in vivo, increasing VGF mRNA levels up to 50-fold in PC12 cells -triphosphate using terminal deoxynucleotidyl transferase . Specificity controls were (1) pretreating sections with RNase A for 1 hr before hybridization and (2) coincubation of the35S-labeled oligonucleotide in the hybridization medium with a 100-fold excess of unlabeled oligonucleotide.eviously using coImmunohistochemical staining was performed on 40 \u03bcm free-floating cryosections of L4/L5 spinal cord. For VGF, the sections were blocked for 1 hr in TTBS containing 3% normal rabbit serum (NRS) at room temperature and incubated at 4\u00b0C for 72 hr with goat \u03b1-VGF diluted 1:5000 in TTBS. Followed after three 10 min washes in 0.1 M phosphate buffer by a 90 min incubation at room temperature with biotinylated rabbit anti-goat secondary antibody diluted 1:200 in TTBS. A further three washes in 0.1 M phosphate buffer were followed by a 60 min incubation at room temperature with Avidin Biotin complex . Signal was amplified using a tyramide amplification protocol. Controls were carried out without the primary antibody. In addition, 3 independent primary antibodies to VGF were used. VGF (R15) which recognises the C-terminus of VGF, VGF (D20) which recognises the N-terminal of VGF and VGF (G17) which also recognises the N-terminus. All three antibodies showed the same clear and consistent increase in staining in the DRG and dorsal horn following SNI despite being raised to three separate VGF epitopes. Ultimately, VGF (G17) was selected as the staining was the clearest.Sections were double labeled for neuronal nuclei 1:5000), calcitonin gene regulated polypeptide 1:4000), ionized binding calcium adaptor molecule-1 1:2000), isolectin B4 ((Sigma) 1:200), 5 HT 1:75).Quantitative real-time PCR (RT-PCR) was performed using the SYBR green detection system with primer sets designed on Primer Express . Specific PCR product amplification was confirmed using dissociation protocol. Transcript regulation was determined using the relative standard curve method per manufacturers' instructions. Relative loading was determined prior to RT-PCR with RNA spectrophotometry followed by gel electrophoresis and post RT-PCR by amplification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). For each time point 4 samples of pooled tissue from 2 rats were analyzed.2) using a 10 \u03bcl 26 G Hamilton syringe. The experimenter was blind to the contents of the syringe and only one sensory modality was tested per rat.Flexion withdrawal reflex thresholds to punctate mechanical stimulation of the plantar surface of the hindpaw were established in na\u00efve adult rats using von Frey filaments (VF) applied sequentially to the plantar surface of the hind paw 10 times at one second intervals. Threshold was defined as the VF filament causing paw 50% withdrawal. Cold responses were measured by placing a drop of acetone on the plantar of the paw and recording the number of seconds that the paw was withdrawn over the following 20 seconds. Rats were tested one day prior to intrathecal injections to establish baseline sensitivity and again immediately before anaesthetising for intrathecal injection with TLQP-62 or vehicle. Mechanical and cold sensitivity were then tested 30 min after application and every 15 minutes for four hours Intrathecal injections were performed at the L4\u20135 level in anesthetized rats . Single spikes with cutaneous receptive fields on the hindpaw were recorded throughout the depth of the dorsal horn and analysed using Chart software . Spontaneous activity was recorded for one minute followed by 3 brush stimuli and 3 pinch stimuli . A single drop of acetone was the applied to the receptive field and responses recorded for 30 seconds. Following baseline recording, 20 ul of TLQP-62 (50 nM) in saline was applied directly onto the exposed spinal cord and stimuli were repeated at 5 and 10 minutes post drug application and every ten minutes thereafter for 60\u2013120 minutes.chniques . Rats we2,1 EGTA, 10 HEPES, 2 MgATP . Superficial dorsal horn neurons were visualized with IR-DIC, and voltage clamp recordings of whole-cell currents were obtained using a Multiclamp 700A amplifier . Spontaneous excitatory post-synaptic potentials (sEPSCs) were observed at a holding potential of -70 mV and baseline activity was established for at least 5 min before drug application. TLQP62 (50 nM in aCSF) was bath applied for up to 30 minutes and its effects on sEPSC amplitude and frequency analysed off-line.Superficial dorsal horn neurons were patched at room temperature in 400 \u03bcm lumbar sagittal slices in 21 day old na\u00efve rats . PipetteBDNF: brain derived neurotrophic factor; CGRP: calcitonin gene related peptide; DLF: dorsal lateral funiculus; DRG: dorsal root ganglia; NGF: nerve growth factor; sEPSC spontaneous excitatory postsynaptic potential; SNI: spared nerve injury.The authors declare that they have no competing interests.in vivo physiological studies. AT performed immunohistochemical studies. LL performed in situ hybridization studies. MB conceived and performed some synaptic physiology experiments. GH contributed to the behavioural investigations as well as preparation of the manuscript. MC performed qPCR and behavioral investigations. SS discovered and provided VGF for this study and aided in the conception of this work. MF conceived the project, lead the experimental design and data analysis and prepared the final manuscript.AM conceived and performed behavioural, histological and quantitiative PCR studies as well as interpreting data and aiding in the preparation of the manuscript. RI conceived and performed synaptic physiology experiments. SK performed Origins of terminal staining of VGF in the lumbar spinal cord. A. Immunohistochemistry of VGF (left) and CGRP (right) in the L4 spinal cord following unilateral L3\u20135 dorsal rhizotomy 5 days earlier. Ipsilateral CGRP depletion indicates total loss of primary afferent input. VGF staining in the region is decreased but not totally depleted (arrow). B. VGF (left) and 5 HT (right) immunostaining in L4 spinal cord following unilateral lesion of the dorsolateral funiculus at upper thoracic level 5 days earlier. Ipsilateral 5-HT depletion indicates loss of descending brainstem terminals, while VGF is only partially diminished (arrow). C Retrograde labeling of projection neurones in the rostroventral medulla, using bilateral True blue (2%) injection into the L4/5 spinal cord under anaesthetic, 5 days earlier (red), demonstrates expression of VGF in brainstem descending projection neurones. True blue:red. VGF:green. Double labeled: orange.Click here for fileOrigins of intrinisic staining of VGF in the lumbar spinal cord. A Immunohistochemistry for VGF and NeuN highlighting the ipsilateral increase in VGF in intrinsic dorsal horn neurones (Arrow). B High-power confocal immunohistochemistry of VGF and markers for microglia (Iba-1), neurones (NeuN) and astrocytes (GFAP) demonstrates that intrinsic VGF protein expression is co-localised specifically with the neuronal marker NeuN but not with either of the glial markers (arrows).Click here for file"} +{"text": "Dopamine and dopamine transporters are implicated in the modulation of attention but their specific roles are not well understood. Here we hypothesized that dopamine modulates attention by facilitation of brain deactivation in the default mode network (DMN). Thus, higher striatal DAT levels, which would result in an enhanced clearance of dopamine and hence weaker dopamine signals, would be associated to lower deactivation in the DMN during an attention task.11C]cocaine used as DAT radiotracer) and brain activation and deactivation during a parametric visual attention task in healthy controls. We show that DAT availability in caudate and putamen had a negative correlation with deactivation in ventral parietal regions of the DMN and a positive correlation with deactivation in a small region in the ventral anterior cingulate gyrus (BA 24/32). With increasing attentional load, DAT in caudate showed a negative correlation with load-related deactivation increases in precuneus.For this purpose we assessed the relationship between DAT in striatum and cingulate gyrus . These findings suggest that the beneficial effects of stimulant medications (increase dopamine by blocking DAT) in inattention reflect in part their ability to facilitate the deactivation of the DMN. Dopamine (DA) neurotransmission is believed to play a central role in attention Positron emission tomography (PET) technology can be used to directly measure markers of DA activity. Likewise, functional magnetic resonance imaging (fMRI) allows mapping of brain activation in brain regions modulated by DA Multiple brain regions have been implicated in attention 11C]cocaine to measure DAT, and BOLD-fMRI to measure brain activation during a parametric visual attention (VA) task To test this hypothesis we used PET and cocaine radiotracer to measure DAT availability in the brain. PET images were acquired in 3D mode using a Siemens HR+ tomograph with 4.5 mm isotropic spatial resolution. Each sequential dynamic scan started immediately after intravenous injection of 4 to 10 mCi of [11C]cocaine (specific activity >0.4 Ci/\u00b5mol at time of injection) for a total of 60 minutes of scanning Subjects underwent PET scans with [11C]cocaine images as previously described Regions of interest (ROI) in the striatum (caudate and putamen) and in the cerebellum were defined directly from the cocaine to measure DAT availability in the striatum, and BOLD-fMRI to evaluate brain activation during a parametric VA task. Increased DAT availability in striatum, which would result in lower DA in the synapse, was associated with lower deactivation of the precuneus (BA 7) and higher deactivation of the ventral ACG (BA 24/32). Also, increased DAT in caudate was associated with less deactivation in precuneus with increasing attentional load. Thus, this study suggests that lower DAT availability , such as that observed in ADHD patients treated with methylphenidate, may facilitate attention by modulating brain deactivation in DMN regions (precuneus).We used PET with [Figure S1Sagital views of the DAT-BOLD correlation patterns during VA tasks with (top panel) and without (bottom panel) the outlier rendered to a structural MRI image . The light-blue circle highlights the thalamic cluster.(3.44 MB TIF)Click here for additional data file.Figure S2BOLD-fMRI activation patterns during VA tasks pattern rendered to a structural MRI image . [First row] Average activation and deactivation across all three difficulty levels ; [second row] increased activation/deactivation caused by increased attentional load . B: Statistical maps of positive (top row) and negative (bottom row) correlations between BOLD-fMRI responses in the brain and [11C]cocaine (DAT) radiotracer binding in the striatum (caudate and putamen), rendered to a structural MRI image. Multiple regression (random-effects) analyses. Color maps are t-score windows: 2.7 to 5.(1.40 MB TIF)Click here for additional data file."} +{"text": "We have studied 30 patients presenting with breast cancer and 36 control patients admitted to hospital for minor surgery. Stool specimens were obtained for bile acid analysis and bacterial nuclear dehydrogenation activity (NDC) estimation. The mean total faecal bile acid (FBA) concentration (mumol/g) in patients with breast cancer was 15.6 /+- 1.8 s.e., significantly lower than for control patients (20.5 /+- 1.9). NDC were isolated from the faeces of 58% of breast cancer patients an 15% of control patients, this difference being statistically highly significant (P less than 0.005). Increased bile-acid degradation by bacteria in the large bowel may explain the reduced FBA concentration in patients with breast cancer. Increased NDC isolation in breast-cancer patients suggests that oestrogen production in the colon may play a role in the aetiogy of breast cancer in some patients."} +{"text": "Congenital hypertrophy of the retinal pigment epithelium (CHRPE) and multiple mandibular osteomata are markers of familial adenomatous polyposis (FAP). We have assessed their prevalence in non-polyposis familial colorectal neoplasia. Multiple mandibular osteomata were present in 1/29 (3%) patients with familial colorectal neoplasia. CHRPE was present in 11/33 (33%) patients with familial colorectal neoplasia compared with 3/36 (8%) with sporadic disease (P = 0.01) and 4/32 (12.5%) control subjects (P = 0.04). Seven patients with familial colorectal neoplasia had multiple areas of CHRPE compared with one with sporadic disease (P = 0.02) and one control subject (P = 0.02). There was no obvious correlation between calculated familial colorectal cancer risk and the presence of multiple areas of CHRPE. A proportion of patients with familial colorectal cancer have a marker found in FAP and may therefore have a constitutional genetic defect, at least in part responsible for their cancer, making them an interesting group for genetic study. Ophthalmoscopy may contribute to risk assessment in familial colorectal cancer."} +{"text": "Lymphoid tissue from 43 cases of canine lymphosarcoma and from 40 clinically normal dogs have been examined for markers of retrovirus infection. From 69-76% of culture supernatants from lymphosarcomas were shown to contain particles of retroviral density and to possess poly rC-oligo dG templated polymerase (reverse transcriptase) activity compared with 17-24% of culture supernatants from normal canine lymphoid cells. In 6 culture supernatants from cases of lymphosarcoma, high molecular weight 60-70S RNA was detected and shown to be found in association with this particulate reverse transcriptase activity. No such RNA was detected in 6 culture supernatants from normal canine lymphoid cells."} +{"text": "The effect of 24-hours continuous somatostatin 14 infusion on the volume of the bile secretion and on thebile composition were studied in seven patients with malignant biliary obstruction who had transhepaticexternal biliary drainage.+, K+, CI\u2212, Ca++ and Mg++. The loss of chloride and sodium was reduced with median 50 mmol/day each(p=0.01). The relative concentrations of the measured bile constituents did not change significantly,except for bile acids (p=0.02): the concentration of glycochenodeoxycholic acid increased significantly(p=0.04). The molar loss of taurocholic acid decreased significantly (p=0.035), so the increasedconcentration of glycochenodeoxycholic acid resulted only in a marginally significant reduction in thetotal molar loss of bile acids (p=0.051).The bile acid composition was measured with high performance liquid chromatography (HPLC).Somatostatin infusion significantly reduced the daily bile loss from median 473 ml to 140 ml with a concomitant significant reduction in the daily molar loss of cholesterol, triglyceride, NaSomatostatin is a potent inhibitor of bile secretion. The peptide may be used in severely bile depletedpatients for reducing their serious electrolyte and acidity problems. Analysis of bile acid composition byHPLC is well suited for further investigations of the regulatory mechanisms of bile acid secretion."} +{"text": "Biopsies from 40 human tumours, obtained at operation, have been transplanted to mice treated with antilymphocyte serum (ALS); viable grafts were obtained from 6 tumours. A further 26 human tumours were transplanted to mice thymectomized as adults and treated with ALS; viable grafts were obtained in 6 cases."} +{"text": "INK4/ARF (CDKN2A/B) tumor suppressor locus with risk of atherosclerotic diseases and type 2 diabetes mellitus. To explore the mechanism of this association, we investigated whether expression of proximate transcripts correlate with genotype of representative 9p21 SNPs.Genome-wide association studies (GWAS) have linked common single nucleotide polymorphisms (SNPs) on chromosome 9p21 near the INK4/ARF locus. Correlations among these variables were determined by univariate and multivariate analysis. Significantly reduced expression of all INK4/ARF transcripts was found in PBTL of individuals harboring a common SNP (rs10757278) associated with increased risk of coronary artery disease, stroke and aortic aneurysm. Expression of MTAP was not influenced by rs10757278 genotype. No association of any these transcripts was noted with five other tested 9p21 SNPs.We analyzed expression of 9p21 transcripts in purified peripheral blood T-cells (PBTL) from 170 healthy donors. Samples were genotyped for six selected disease-related SNPs spanning the INK4/ARF locus, which encodes three related anti-proliferative transcripts of known importance in tumor suppression and aging.Genotypes of rs10757278 linked to increased risk of atherosclerotic diseases are also associated with decreased expression in PBTL of the INK4/ARF (CDKN2A/B) tumor suppressor locus on chromosome 9p21 correlate with risk of atherosclerotic diseases (coronary artery disease (CAD) INK4a, one of the proteins encoded by the INK4/ARF locus, controls pancreatic islet proliferation and mass with aging Recent human genome-wide association studies (GWAS) have shown that SNPs with high minor allele frequencies near the INK4/ARF locus , indicating a potentially important role for cell cycle inhibitors in atherosclerotic disease.In this work, we sought to determine whether there is a link between these disease-associated SNPs and expression of 9p21 transcripts. We found that individuals harboring a common SNP genotype associated with increased risk for atherosclerotic disease demonstrated markedly reduced expression of This study was approved by the University of North Carolina Institutional Review Board with all participants providing informed written consent.2-transformed expression . In brief, PBTLs were obtained from 170 healthy subjects and isolated to >90% purity by Magnetic Activated Cell Sorting (MACS). MACS, RNA preparation and transcript expression analysis were performed in PBTL as described INK4/ARF transcript expression (log2-transformed or \u0394\u0394CT) and rs10757278 genotype (We selected six SNPs for genotyping based on strength of association in previous studies: two associated with atherosclerosis (rs10757278 and rs518394) genotype . This SNARF, INK4bp15, INK4ap16, and ANRIL in GG vs. AA individuals were also significantly increased in AA individuals compared to GG individuals was not associated with rs10757278 genotype are well-described at other loci Here, we show that the expression of INK4/ARF expression and atherogenesis.It is important to note that rs10757278 and linked SNPs in high LD (e.g. rs1333049) are associated with family history of atherosclerotic disease, but not other traditional risk factors including body mass index, diabetes, tobacco use, blood pressure, C-reactive protein or lipid levels INK4/ARF transcripts might modulate the earliest stages of atherosclerosis. The INK4-class cyclin dependent kinase (CDK) inhibitors, of which p15INK4b and p16INK4a are founding members, block cell cycle progression by inhibiting the activity of the proliferative kinases CDK4 and CDK6. A role for cell cycle inhibitors has been postulated in promoting favorable vascular remodeling and preventing pathologic intimal hyperplasia by regulating G1 to S phase progression in vascular smooth muscle cells complexes potently repress INK4/ARF locus expression Xist at the inactive X-chromosome, reviewed in ANRIL expression could potentially regulate INK4/ARF repression through PcG complexes, explaining why all four INK4/ARF transcripts are coordinately associated with rs10757278 genotype. On the other hand, ANRIL could also influence the expression of other genes in trans- as has been recently reported for HOTAIR at Hox loci As the putative non-coding RNA INK4/ARF expression on the development of atherosclerosis do not result from their expression in PBTL, but rather, observations in PBTL may serve as a surrogate for INK4/ARF expression in other tissues. As cis-regulatory elements may act as enhancers in one tissue and repressors in another, it is also possible that genetic variants in LD with rs10757278 genotype might be associated with increased INK4/ARF transcript expression in other tissues of greater relevance to atherogenesis . Several investigators have argued that cellular senescence, which is induced by p16INK4a, promotes atherosclerosis Click here for additional data file.Table S2(2.42 MB TIF)Click here for additional data file."} +{"text": "Rhizobium leguminosarum bv. viciae mutants unable to transport branched-chain amino acids via the two main amino acid ABC transport complexes AapJQMP and BraDEFGC produce a nitrogen starvation phenotype when inoculated on pea (Pisum sativum) plants 2 fixation on peas demonstrating that a low rate of branched amino acid transport in R. leguminosarum bv. viciae supports wild-type rates of nitrogen fixation. The importance of branched-chain amino acid transport was then examined in other legume-Rhizobium symbioses. An aap bra mutant of R. leguminosarum bv. phaseoli also showed nitrogen starvation symptoms when inoculated on French bean (Phaseolus vulgaris), a plant producing determinate nodules. The phenotype is different from that observed on pea and is accompanied by reduced nodule numbers and nitrogen fixation per nodule. However, an aap bra double mutant of Sinorhizobium meliloti 2011 showed no phenotype on alfalfa (Medicago sativa).A region important in solute specificity was identified in AapQ and changing P144D in this region reduced branched-chain amino acid transport to a very low rate. Strains carrying P144D were still fully effective for NSinorhizobium meliloti-alfalfa symbiosis under the conditions measured. The contrasting symbiotic phenotypes of aap bra mutants inoculated on different legumes probably reflects altered timing of amino acid availability, development of symbiotic auxotrophy and nodule developmental programmes.Symbiotic auxotrophy occurs in both determinate pea and indeterminate bean nodules demonstrating its importance for bacteroid formation and nodule function in legumes with different developmental programmes. However, only small quantities of branched chain amino acids are needed and symbiotic auxotrophy did not occur in the These symbioses arise from infection of host plants, mainly of the legume family, by proteobacteria and results in root structures called nodules Papilionoideae being indeterminate with a cylindrical shape and a persistent meristem at the tip of the nodule. Bacteria inside symbiosomes differentiate into N2 fixing bacteroids and this is accompanied by dramatic increases in size, shape and DNA content Phaseolus, Vigna or Glycine), form spherical determinate nodules with a transient meristem; bacteria do not undergo endoreduplication and therefore do not enlarge substantially. These bacteroids retain a normal DNA content and are largely viable after isolation from nodules The largest input of available nitrogen into the biosphere comes from biological reduction of atmospheric NPisum sativum) nodules R. leguminosarum bv. viciae strains contain two broad specificity amino acid ABC (ATP binding cassette) transporters, AapJQMP and BraDEFGC. AapJQMP consists of a solute binding protein (SBP) AapJ, two permease units AapQM and an ABC subunit dimer formed by AapP for ATP hydrolysis. The BraDEFGC complex consists of SBP BraC, permeases BraDE and ABC subunits BraFG. The aap bra double mutants, RU1357 and RU1722, formed pea bacteroids that appeared morphologically normal in electron micrographs and fixed nitrogen per bacteroid at wild-type levels, but the plants were nitrogen starved aap bra mutant bacteroid protein levels per plant were reduced to \u223c30% of wild-type. The reason for the nitrogen starved phenotype was later identified to be a limitation of branched-chain amino acid biosynthesis by developing bacteroids The shared metabolism between the symbiotic partners has been proposed to be a simple nutrient exchange of dicarboxylates for ammonium in the nodule R. leguminosarum bv. viciae strains in pea nodules. This shows that large quantities of amino acid are not needed and is consistent with symbiotic auxotrophy. Additionally we report the symbiotic phenotypes of an aap bra mutant in R. leguminosarum bv. phaseoli inoculated on French bean (Phaseolus vulgaris) forming determinate nodules and Sinorhizobium meliloti on alfalfa (Medicago sativa) forming indeterminate nodules.Here we provide evidence that only a low rate of branched-chain amino acid transport is required to overcome symbiotic auxotrophy of aap bra null mutant RU1722 (data not shown). Of these mutants P144D proved most interesting, so a single copy of AapQ P144D was generated by recombination into the chromosome of Rlv3841. This was achieved by replacing the \u03a9Sp cartridge in RU1722 aapJQM::\u03a9Sp braEF::\u03a9Tc with a full copy of the aapJQM operon containing the mutated AapQ P144D, generating RU1976. The braEF::\u03a9Tc null mutation is present in all the control strains so transport occurs exclusively via Aap. The uptake properties of RU1976 (aapQ P144D braEF::\u03a9Tc) were compared to those of RU1721 (braEF::\u03a9Tc), which contains a wild-type copy of Aap and the aap bra deletion mutant RU1722 (aapJQM::\u03a9Sp braEF::\u03a9Tc) (max) of transport of small amino acids such as alanine was less severely affected (51% of control) by the AapQ P144D mutation than transport of larger amino acids such as glutamate (25% of control). The lower levels of amino acid uptake observed in RU1976 could also arise from a reduction in solute affinity (Km). In order to investigate this, uptake rates were determined for glutamate, alanine, aspartate and leucine at varying concentrations in both RU1721 and RU1976. These were then used to calculate Km for amino acid uptake by these strains such as AapQ and AapM might differentially affect the uptake of amino acids EF::\u03a9Tc) . The max strains . It is cno acids .2-fixation and presumably bacteroid formation. Consistent with the proposal of symbiotic auxotrophy only a low rate of branched-chain amino acid transport is needed for N2-fixation.In order to analyse if reduced transport by Aap triggers symbiotic auxotrophy, strain RU1976 and the control strains, RU1721 and RU1722 were inoculated onto pea plants. Dry weights of 6 week old pea plants inoculated with RU1976 and RU1721 were similar, while RU1722 dry weights were reduced as expected . Thus a R. leguminosarum bv. viciae strain A34 and its aapJQM::\u03a9Sp braE::TnphoA mutant RU1357, have been previously described R. leguminosarum bv. phaseoli 4292 and R. leguminosarum bv. viciae A34 are isogenic strains that only carry different symbiotic (sym) plasmids. Strain 4292 nodulates Phaseolus while strain A34 nodulates Pisum, Vicia and LensaapJQM::\u03a9Sp (RU1809), braE::TnphoA (RU1932) and aapJQM::\u03a9Sp braE::TnphoA (RU1933) were isolated in R. leguminosarum bv. phaseoli 4292 as described in R. leguminosarum bv viciae A34 and R. leguminosarum bv. phaseoli 4292 (aap bra double mutant (RU1933) the transport of all solutes was reduced to background levels, except for alanine and histidine which under the assay conditions can still enter the cells via other transport systems R. leguminosarum bv. viciae double transport mutants on pea plants R. leguminosarum bv. viciae cells are dependent on the provision of branched-chain amino acids by the host plant to develop into mature bacteroids and probably also for their persistence The transport specificities and symbiotic phenotypes of oli 4292 . Thus inP. vulgaris) was measured at different time points. Plants at 8 weeks showed no significant differences when inoculated with the wild-type (RU2222), the aap mutant (RU1809) and the bra mutant (RU1932) reduced dry weights compared to wild-type RU2222 inoculated plants. Plants inoculated with RU1933 also showed signs of yellowing after 8 weeks although not as severe as uninoculated controls (aap bra mutants of R. leguminosarum bv. viciaeGrowth and symbiotic performance of French bean ((RU1932) . Howevercontrols . NitrogeAt 5 and 6 weeks the reduced symbiotic performance of strain RU1933 can be eaap bra mutant RU1933 showed remarkable differences (aap bra mutant background RU1933 (TEM pictures of bacteroids formed by wild-type RU2222 and the ferences . Bacterod RU1933 . TEM micS. meliloti livHMGFK is homologous to R. leguminosarum braDEFGCS. meliloti livK as braC and livH as braD. The double mutants aapJ::\u03a9Sp braC::mTn5 (LMB210) and aapJ::\u03a9Sp braD::mTn5 (LMB211) were generated in the S. meliloti 2011 background as described in aapJ braD mutant LMB211, which is very similar to R. leguminosarum strains is able to interact with the Bra membrane complex and is very specific for the branched-chain amino acids and alanine S. meliloti strain1021. strains . Thus muMedicago sativa plants inoculated with wild-type 2011 or the aapJ braD double mutant LMB211 were analysed for nitrogen fixation (AR) and dry weights after 4 weeks of growth. Neither nitrogen fixation rates nor dry weights differed significantly (p>0.05) between S. meliloti 2011 and LMB211, although uninoculated control plants showed strong signs of nitrogen starvation and a corresponding drop in dry weights ( weights .aapJ braD mutant LMB211 bacteroids showed no obvious differences in development (data not shown) and flow cytometric characterisation of bacteroids also failed to demonstrate developmental alterations of bacteroids formed by LMB211 (data not shown).TEM pictures of wild-type 2011 and the Rhizobium leguminosarum bv. viciae strains mutated in the two broad specificity amino acid ABC uptake systems, Aap and Bra develop a severe nitrogen starvation phenotype braC was mutated in an aap null mutant background and it was shown that strain RU1979 (\u0394aapJ braC::\u03a9Spec) retains the transport of branched-chain amino acids. This is because of the presence of an orphan SBP (BraC3) which specifically binds branched-chain amino acids and interacts with the core Bra membrane complex. Strain RU1979, which only retains branched-chain amino acid transport, fixes nitrogen at wild-type rates on pea plants. In addition complementation analysis using heterologous ABC transport systems demonstrated that branched-chain amino acid transport alone rescues the starvation phenotype on pea plants. Microarray analysis of pea bacteroids showed that the pathways of branched-chain amino acid biosynthesis are transcriptionally down regulated leading to premature arrest of development in aap bra bacteroids Pea plants inoculated with 2-fixation. It was already known that the single aap and bra mutants have no symbiotic phenotype so the wild-type rate of transport can be lowered by \u223c50% with no effect aapQ P144D braEF::\u03a9Tc) were 1\u20132 nmol min\u22121 mg protein\u22121, while the aap bra double mutant strains have rates below 1 nmol min\u22121 mg protein\u22121. This rate for RU1976 is roughly 10% of the maximum rates of branched-chain amino acid uptake measured in wild-type R. leguminosarum bacteroids are symbiotic auxotrophs and do not require amino acids for catabolism, which would need a much higher rate of transport.Our first aim in this study was to determine how much branched-chain amino acid transport is needed for fully effective NleuD mutant of R. leguminosarum strain 3841 required leucine at 100 \u00b5M added to AMS glucose/NH4Cl cultures to overcome auxotrophy and restore a wild-type rate of growth. By comparison as sole nitrogen source 10 mM leucine was needed to obtain the same growth rate as NH4Cl.A S. meliloti in symbiosis with alfalfa (M. sativa) and Bradyrhizobium japonicum with soybean (Glycine max) R. leguminosarum bv. phaseoli and French bean. This was done for two reasons. First, beans have determinate nodules, while peas have indeterminate nodules. Second, R. leguminosarum bv. viciae A34 and R. leguminosarum bv. phaseoli 4292 differ only in their Sym plasmid so the same aap bra mutations can be directly compared in pea and bean hosts.Published microarray data shows transcriptional down regulation of branched-chain biosynthetic genes occurs in aap bra) became nitrogen starved, indicated by reduced dry weight, nitrogen fixation, and nitrogen fixation per nodule as well as yellowing and increased PHB accumulation in bacteroids. Unexpectedly, they produced fewer nodules on beans, in contrast to the increase in nodule formation on peas by equivalent aap bra mutants. Legumes usually increase nodule numbers when inoculated with fix reduced strains. The reduction in French bean nodules inoculated with RU1933 might result from limitation of branched-chain amino acids during infection or very early in bacteroid development leading to reduced success in nodule formation. Therefore, while peas and beans have different nodulation responses to aap bra mutants their different patterns of nodule development probably alter the timing of amino acid availability and the development of symbiotic auxotrophy.French bean plants inoculated with RU1933 4Cl as nitrogen source. S. meliloti was grown in M9 medium with 1.25 mM CaCl2, 2.5 mM MgSO4, 0.1 ug/ml Biotin, 10 ug/ml CoCl2, 10 mM D-glucose and 10 mM NH4Cl. Antibiotics were used at the following concentrations (\u00b5g ml\u22121): streptomycin (St), 500; neomycin (Nm), 80; tetracycline (Tc), 5; gentamicin (Gm), 20 and spectinomycin (Sp), 100.The bacterial strains, plasmids and primers used in this study are detailed in Rhizobium leguminosarum bv. viciae AapQ site directed mutant was generated as follows. AapJQMP was amplified using primers p495 and p441. The 5 kb PCR product was TOPO cloned into pENTR/D-Topo and the CCG triplet coding for proline at position 144 within AapQ replaced by a GAC coding for aspartic acid (P144D) using the Quick Change site-directed mutagenesis kit . The region was transferred by a Gateway LR reaction into the pJQ200SK derivative, pGW1, producing pRU1247. This plasmid was conjugated into RU1722 (aapJQM::\u03a9Sp braEF::\u03a9Tc) to generate the homogenote RU1976 by sucrose selection and loss of the spectinomycin marker. The correct mutation was confirmed by PCR.The Rhizobium leguminosarum bv. phaseoli mutants were generated as follows. Plasmid pRU410 MfeI deletion (3.2 kb) spanning aapJQM, was transformed into Escherichia coli S17-1 and conjugated into R. leguminosarum bv. phaseoli 4292. This plasmid is derived from pJQ200SK sac genes. Transconjugants were spread on AMS agar plates aapJQM::\u03a9Sp genotype by PCR mapping using primers p418 and p419. As expected from the phenotype of aap mutants of R. leguminosarum bv. viciae, strain RU1809 did not grow on minimal medium with L-glutamate as the sole carbon and nitrogen source. To generate bra mutants, cosmid pBIO206A braE::TnphoA) was transformed into S17-1 and conjugated into strain 4292 and RU1809. Colonies were selected for tetracycline and kanamycin, resistance, before the chaser plasmid pPH1JI (gentamicin resistant) was then conjugated into them to allow the selection of recombinants with loss of the tetracycline marker and resistance to kanamycin and gentamicin. Plasmid pPH1JI was then removed by conjugating pLAFR1 into all strains, and selecting for loss of the gentamicin marker while retaining resistance to kanamycin and tetracycline. The braE::TnphoA insertion was confirmed by PCR mapping using primers pIS50R and p439. The braE::TnphoA single and aapJQM::\u03a9Sp braE::TnphoA double mutants were designated as RU1932 and RU1933, respectively. Strain 4292 containing pLAFR1 was designated RU2222.Sinorhizobium meliloti mutants were generated as follows. A series of mTn5STM mutants in the livHMGFK operon of S. meliloti strain 2011 S. meliloti livHMGFK is homologues to the R. leguminosarum braDEFGC operon livK (braC) mutant 2011mTn5STM.3.12.F11 and a livH (braD) mutant 2011mTn5STM.4.05.D04 were chosen as a background for an aapJ insertion mutation using an \u03a9Sp cassette S. meliloti strain 2011 aapJ gene. The resulting PCR product was cloned into pJET1.2 resulting in plasmid pLMB111. Primers pr0165 and pr0166 were used in an inverse PCR to amplify the aapJ surrounding DNA from pLMB111 without the aapJ ORF. The resulting PCR product was digested with BamHI (underlined in primers) and religated forming plasmid pLMB112. An XbaI/XhoI fragment was then cloned from pLMB112 into pJQ200SK BamHI and a \u03a9Sp cassette was inserted producing pLMB118. This plasmid was used to generate aapJ::\u03a9Sp homogenotes in the braC and braD mutant backgrounds to produce strains LMB210 and LMB211, respectively. The insertion was mapped from both sides using primers pOTforward and pr0240 or primer pr0241, respectively.R. leguminosarum uptake assays were performed with 25 \u00b5M (4.625 kBq of 14C) solute 4Cl to an OD600 of \u223c0.4. S. meliloti was grown in modified M9 medium with 10 mM D-glucose and 10 mM NH4Cl to an OD600 of \u223c0.4.Phaseolus vulgaris cv. Tendergreen) were surface sterilised and grown for 5 to 6 weeks in a sterile mixture of fine gravel and vermiculite (1\u22364) in 15 l pots with 7 seeds per pot in a controlled environment at 22\u00b0C and a 8:16 h day: night cycle as described earlier Medicago sativa cv. Europe plants were grown in 1 l pots in vermiculite with 7 seeds per pot for 4 weeks watered with nitrogen free rooting solution French bean plants (Acetylene reduction was measured as previously described Bean shoot dry weights were determined after 8 weeks, pea shoot dry weights after 6 weeks and alfalfa shoot dry weights after 4 weeks of growth.After counting and removing all nodules from the plants, nodules were crushed in 40 mM HEPES buffer and bacteroids harvested by differential centrifugation S. meliloti bacteroids was performed as described elsewhere Flow cytometric analysis of"} +{"text": "Calluna vulgarts). This terpene was found toinhibit HL-60 leukaemic cell proliferation and arachidonic acidoxidative metabolism in various cell species. The effects of ursolicacid and its analogues on soybean 15-lipoxygenase activity and onthe proliferation of a human gastric tumour cell line (HGT), havebeen assessed. These triterpenes inhibited soybean 15-lipoxygenaseat its optimal activity (pH 9). The proliferation ofHGT wasdecreased in a dose-dependent manner. At 20 \u03bcM the rank order is:ursolic acid > uvaol > oleanolic acid > methyl ursolate. Thecarboxylic group at the C28 position of ursolic acid appears to beimplicated in the inhibition of both lipoxygenase activity and cellproliferation. Thus methylation of this group decreases these twoinhibitory properties. Oleanolic acid, which differs by the positionof one methyl group (C20 instead of C19) is less inhibitory thanursolic acid. The lipophilicity of the terpene is also implicatedsince uvaol appears to be more inhibitory than methyl ursolate.The authors have previously isolated and purified ursolic acid fromheather flowers ("} +{"text": "Blood glucose (glc) was thenmeasured weekly for 16 weeks. Data showed thatthe experimental group contained 70% euglycemicanimals (defined as glc <200mg/dL) versus 10% ofthe control animals (P<.05) at 14 weeks. Meanweight in the treated group was greater than theuntreated group. Insulin mRNA was detected at theinjection site of all of the treated animals, but notcontrols. Complete destruction of islets was confirmedby histology ruling out the possibility ofspontaneous reversal of insulinitis. We concludethat IM delivery of the insulin gene in the NODmouse was able to prevent clinical DM up to 14weeks in a majority of treated animals. Our experimentaldata suggests that gene therapy may be analternative treatment for IDDM in the future.Using the Adeno-associated virus (AAV) as a genedelivery vehicle, we have constructed a recombinantvector containing the full length rat preproinsulingene (vLP-1). Utilizing the well described non-obesediabetic (NOD) mouse model, an experimentalgroup (n=10) of animals were intramuscularly (IM)injected with 10"} +{"text": "To confirm several recent studies pointing to loss of heterozygosity (LOH) at BRCA2 as a prognostic factor in sporadic breast cancer, we examined this genetic alteration in a large series of human primary breast tumours for which long-term patient outcomes were known. LOH at BRCA2 correlated only with low oestrogen and progesterone receptor content. Univariate analysis of metastasis-free survival and overall survival (log-rank test) showed no link with BRCA2 status . LOH at BRCA2 does not therefore appear to be a major prognostic marker in sporadic breast cancer."} +{"text": "Objective: The objectives of this study were to 1) determine the prevalance and characterize the symptomatology of Trichomonas vaginalis (TV) infection in pregnant women on entry into prenatal care in an inner-city population; 2) compare conventional microscopic methods vs. culture techniques in diagnosing TV in both symptomatic and asymptomatic pregnant patients; and 3) correlate wet mount microscopic and microbiologic characteristics of varying manifestations of trichomoniasis. Methods: One thousand two hundred sixty patients in an inner-city population were tested at entry into prenatal care for TV by saline wet mount and culture techniques. Other tests for lower genital tract infection were also performed. Vaginal symptoms were ascertained through standardized questioning prior to examination. Standard microscopic and microbiologic data were also obtained for analysis. Wet mounts were systematically examined and considered negative if no TV was identified in 10 high powerfields (HPFs). Cultures were inspected from days 4 to 7 or until positive results were obtained. Results were analyzed using McNemar's test for correlated proportions, chi-squared test, or Fisher exact test where appropriate. Results: Culture and wet mount results were available in 1,175 patients. TV infection was documented by one or both techniques in 110/1,175 (9.4%). Culture methods detected 105/110 (94.5%) of all patients while wet mount detected 90/110 (73%) (P <0.001). Vaginal symptoms were present in only 20/110 patents (18.2%). Among asymptomatic patients, culture detected 94% while wet mount detected 70% (P < 0.001). Among symptomatic patients, wet mount and culture were both effective and diagnosed 85% and 95% of infections, respectively (P = not significant). Patients with TV were more likely to have increased vaginal fluid wlaite blood cells (WBCs) and more severe vaginal flora disruption than uninfected controls. Subgroup analysis revealed wet mount-positive/culture-positive patients were more likely to have vaginal flora disruption, as evidenced by decreased lactobacilli and elevated vaginal pH, than wet mount-negative/culture-positive subjects. Coexistent infection rates were similar regardless of wet mount status. Elevated vaginal fluid WBCs were more common among patients with symptoms. Conclusions: 1) Screening pregnant women for TV based solely on symptomatology is ineffective in this population; 2) culture techniques detected more infections than conventional microscopic evaluation; and 3) significant increases in vaginal fluid WBCs and altered vaginal flora are found in both symptomatic and asymptomatic TV, suggesting that both infestations have the potential to adversely affect pregnancy outcome. Studies on the influence of TV on pregnancy outcomes are ongoing."} +{"text": "Intrapericardial instillation of 185\u2013370 MBq (5\u201310 mCi) 32P-colloid in 36 patients with malignant pericardial effusion resulted in a complete remission rate of 94.5% (34 patients) whereas two patients did not respond to treatment due to a foudroyant formation of pericardial fluid. The median duration time was 8 months. No side-effects were observed. These results suggest that intrapericardial instillation of 32P-colloid is a simple, reliable and safe treatment strategy for patients with malignant pericardial effusions. Therefore, since further evidence is provided that 32P-colloid is significantly more effective than external radiation or non-radioactive sclerosing agents, this treatment modality should be considered for the management of malignant pericardial effusion. \u00a9 1999 Cancer Research CampaignMalignant pericardial effusion is usually treated only when signs of cardiac tamponade develop. Several methods of treatment have been reported with an overall response rate of approximately 75%. Since our initial study using intrapericardial"} +{"text": "Sno mutants in flies and mice have proven enigmatic in revealing developmental roles for Sno proteins. Thus, to identify developmental roles for dSno we first reconciled conflicting data on the lethality of dSno mutations. Then we conducted analyses of wing development in dSno loss of function genotypes. These studies revealed ectopic margin bristles and ectopic campaniform sensilla in the anterior compartment of the wing blade suggesting that dSno functions to antagonize Wingless (Wg) signaling. A subsequent series of gain of function analyses yielded the opposite phenotype (loss of bristles and sensilla) and further suggested that dSno antagonizes Wg signal transduction in target cells. To date Sno family proteins have not been reported to influence the Wg pathway during development in any species. Overall our data suggest that dSno functions as a tissue-specific component of the Wg signaling pathway with modest antagonistic activity under normal conditions but capable of blocking significant levels of extraneous Wg, a role that may be conserved in vertebrates.The Sno oncogene (Snoo or dSno in Drosophila) is a highly conserved protein and a well-established antagonist of Transforming Growth Factor-\u03b2 signaling in overexpression assays. However, analyses of Transforming Growth Factor-\u03b2 (TGF-\u03b2) family members perform essential tasks during development in all animals more complex than sponges ski-related novel gene) protein shares significant amino acid identity with the viral oncogene v-ski and Sno overexpression causes transformation of chick embryo fibroblasts. Sno is present as a single copy in the human genome but multiple promoters and alternative splicing generate six distinct transcripts. Four isoforms of the Sno protein have been identified with the longest isoform known as SnoN. In cancer, high levels of SnoN are correlated with poor outcome in estrogen-receptor positive breast tumors and gene amplification at the Sno locus is associated with squamous cell carcinoma of the esophagus. Mechanistic studies in mammalian cells revealed that SnoN, as part of a histone deacetylase complex, binds to Smad4 and blocks its ability to transduce TGF-\u03b2 signals. As a result, Sno proteins were initially thought to be obligate antagonists of TGF-\u03b2 signaling The vertebrate Sno . Biochemical studies revealed that Medea - dSno complexes have reduced affinity for Mad and increased affinity for dSmad2 such that in the presence of dSno, Activin signaling is stimulated and Dpp signaling is reduced. The possibility that Sno proteins function as pathway switches in mammals is supported by data that SnoN facilitates Activin signaling in lung epithelial cells and cerebellar neurons Our analysis in Drosophila suggested that Sno has a subtler role in TGF-\u03b2 signaling - as a pathway switch. We found that overexpression of dSno resulted in small wings with multiple vein truncations suggesting antagonism for TGF-\u03b2 family members in the Decapentaplegic/Bone Morphogenetic Protein (Dpp/BMP) subfamily. Alternatively, Sno mutants in both flies and mice have proven enigmatic in revealing developmental roles for Sno proteins, particularly with regard to any requirement for viability. One study of SnoN knockout mice reports early embryonic lethality for homozygous mutant embryos SnoN mutants are viable and that these mice have a defect in T-cell activation dSno mutations are homozygous lethal at the larval/pupal transition and that the lethality is rescued to adulthood by expression of UAS.dSno dSno mutations could survive to adulthood Surprisingly, studies of dSno mutants. Then we conducted loss of function studies utilizing dSno mutants and mutant clones paired with gain of function experiments employing Gal4 driven UAS.dSno. When these paired experiments generated complementary results it increased our confidence that the phenotypes revealed a true role for dSno. We found that dSno restricts Wingless (Wg) signaling in wing imaginal disks. Further we found that dSno accomplishes this by antagonizing Wg signal transduction in target cells. Overall our data suggest that dSno functions as a tissue-specific protein in Wg signaling with modest inhibiting activity under normal conditions but that can effectively block ectopic Wg signals.To gain insight into dSno's role in development we first reconciled the conflicting data on the lethality of dSno is broadly expressed in the wing pouch when compared to the narrow stripe of dpp expression dSno mutant clones would result in Dpp overexpression phenotypes such as those seen with UAS.Mad or UAS.Medea - ectopic veins and enlarged wings.Numerous studies have found that overexpression of dSno results in small wings with multiple vein truncations suggesting that dSno is capable of blocking Dpp/BMP subfamily signaling dSno excision mutants Ex17BdSno and Ex4BdSno all of the reported dSno mutants are likely allelic, 2) the extent of viability for dSno homozygous deletions varies between laboratories due to environmental factors, and 3) a developmental role for dSno is to facilitate Activin signaling during optic lobe development as we reported previously Prior to initiating studies of somatic clones we further characterized the homozygous lethal dSnoEx4B . DNA seqdSnoEx4B and RNA dSnoEx4B revealedty tests with dSnEx4BdSno, Ex17BdSno or sh1402dSno in adult wings (zeste white3 mutant clones (M11zw3) 174dSno homozygous escapers and found they display ectopic margin bristles in the anterior compartment . Under nonsignaling conditions Zw3 phosphorylates Arm tagging it for destruction. Upon receipt of a Wg signal Arm is released from the complex, enters the nucleus and partners with transcription factors to activate gene expression The similarity of the wing phenotypes for Ex17BdSno or sh1402dSno in third instar larval wing disks.Results with both alleles were consistent and those of Ex17BdSno are shown. We examined the expression of Achaete (Ac), a target of Wg signaling in sensory organ precursor cells that will become bristles on the dorsal and ventral surfaces of the anterior wing margin dSno RNA in situ data dSno expression. We found that dSno clones do not affect normal Ac expression but they generate ectopic Ac on the presumptive wing blade . We conclude that dSno does not regulate Wg expression nor the expression of Zw3 (data not shown) and that the effect of dSno mutant clones on Ac is due to a role in restricting Wg signal transduction.To eliminate the possibility that ectopic Ac resulted from alterations in Wg expression we then stained wing disks bearing marked dy to Wg . This exOur first gain of function experiment was designed to determine if dSno was capable of sufficient antagonism for Wg signaling to overcome overexpression of the Wg pathway signal transducer Dsh. For these analyses we employed the wing-specific MS1096.Gal4, a homozygous viable insertion in the second intron of the Beadex gene on the X chromosome. Evidence that MS1096.Gal4 is exclusive to the wing derives from the two reports: complete deletion of the Beadex locus results only in wing defects arm mutant clones . These flies have small and veinless wings (n\u200a=\u200a102), as expected due to antagonism of Dpp signaling . These wes arm4; . The simsensilla and 5G.When we expressed UAS.Dsh with MS1096.Gal4 we found near-absolute lethality . The wings of rare escapers lack surface adhesion, veins and an obvious wing margin. Instead they display a \u201clawn\u201d of ectopic bristles on both wing surfaces. . In thisCoexpression of UAS.dSno and UAS.Dsh with MS1096.Gal4 resulted in nearly complete rescue of lethality with 90.8% of the expected experimental flies observed (n\u200a=\u200a564). The wings (n\u200a=\u200a40) of rescued flies are smaller than UAS.dSno wings and also have no veins . The numWe then tested the Wg antagonism hypothesis molecularly by examining gene expression in third instar wing disks. MS1096.Gal4 expression of UAS.dSno led to a modest reduction in the expression in disks n\u200a=\u200a7; of two WHowever Dsh has been reported to function as a positive factor in the Wg pathway and as a negative factor in Notch signaling in wing disks where Notch is also required for margin bristle development We then examined the expression of Ac (Wg target) and Cut (Notch target) To be certain that dSno does not play any role in Notch signaling in wing development we conducted coexpression experiments with dominant negative forms of both Notch (UAS.DN-Notch) and the Notch pathway transcription factor Mastermind (UAS.MamN). When expressed with MS1096.Gal4, UAS.DN-Notch leads to significant lethality with 12% of the expected experimental flies observed . These wings (n\u200a=\u200a34) are small, have no veins and very few anterior margin bristles . UAS.MamCoexpressing UAS.dSno and UAS.DN-Notch generates additional lethality with only 4.0% of expected adults observed . These wings (n\u200a=\u200a45) display additive effects of each gene's overexpression. Coexpressing wings are smaller than either parental wing, veinless and have lost all their margin bristles . CoexpreAn examination of wing disks also indicates that dSno coexpression does not rescue but rather exacerbates phenotypes due to UAS.DN-Notch and UAS.MamN. UAS.DN-Notch expressing disks have little Ac or Sens expression n\u200a=\u200a7; . UAS.dSnEX17BdSno and sh1402dSno mutant clones. This analysis showed that dSno clones have no effect on Notch, Delta or Serrate expression (data not shown). Taken together the negative results from our examination of interactions between dSno and the Notch pathway lend support to the hypothesis that a normal role for dSno is the restriction of Wg signal transduction during wing development.We also examined the expression of antibodies to the Notch intracellular domain and to the Notch ligands Delta and Serrate in wing disks with S10S10 with MS1096.Gal4 is not quite as lethal as UAS.Dsh - 4.6% of the expected number of adults was observed . These wings bear the hallmarks of ectopic Wg signaling. UAS.ArmS10 wings (n\u200a=\u200a52) lack surface adhesion, are veinless and display numerous ectopic margin bristles on both the dorsal and ventral surfaces . The surviving UAS.dSno, UAS.ArmS10 flies (n\u200a=\u200a262) display similarities and differences from UAS.ArmS10 wings. Wings from UAS.dSno, UAS.ArmS10 flies are smaller and the ectopic bristle phenotype is completely suppressed on the dorsal surface. However, they still display ectopic bristles on the ventral surface and multiple rows of tightly packed stout mechanosensory bristles on the anterior margin with other rows of bristles absent and suppression of the ectopic bristle phenotype. Further the coexpressing UAS.dSno, UAS.Zw3-DN wings (n\u200a=\u200a40) now display a distinct row of margin bristles though its content is mixed or a constitutively active form of Zw3 (Zw3-CA has an S9A mutation in an inhibiting phospho-serine) is mixed . The resAlternatively, MS1096.Gal4 driven UAS.Zw3-CA results in modestly reduced Wg signaling with little lethality - 91% of the expected number of adults was observed . Adults of this genotype have wings (n\u200a=\u200a24) that are smaller than wild type and have no veins. The row of stout mechanosensory bristles on the margin is sparse compared to wild type and there are no ectopic margin bristles . The UASSubsequently we analyzed wings coexpressing dSno and dAxin or dSno and dAxin\u0394RGS Coexpressing UAS.dSno and UAS.dAxin does not generate any lethality but the wing phenotype is enhanced . These wdSno has any role during embryonic development preliminary data suggests that dSno blocks Wg signaling in the ventral epidermis was previously shown to restrict Wg signaling in two embryonic tissues, the midgut and the ventral epidermis. Brk accomplishes this via repressor complexes containing Teashirt that compete for enhancer binding sites with Arm/dTCF activation complexes pidermis . Thus, wExpression of UAS.Brk with MS1096.Gal4 did not generate any lethality . Adult wings (n\u200a=\u200a40) were small, veinless and displayed a novel margin bristle phenotype. No normal margin bristles were evident but instead there were numerous ectopic bristles that appear similar to the pair of large bristles found normally on the margin at the distal tip of the costa . CoexpreMolecular and genetic analyses of phenotypes generated in complementation tests with dSno alleles from three different laboratories reveal that they are alleles of the same gene. These studies also support our previous data that a developmental role for dSno is to facilitate Activin signaling during optic lobe formation in the third instar larval brain. Here via a series of assays we report that another developmental role for dSno is to spatially restrict Wg signaling in third instar larval wing disks. To date TGF-\u03b2-independent functions for mammalian SnoN have been identified in myoblasts dSno mutant clones cell-autonomously express the Wg target gene Ac on the wing blade but have no effect on normal Ac expression suggesting a role for dSno in antagonizing ectopic Wg signaling. Analysis of Wg expression in these clones eliminated the possibility that loss of dSno affects the transcription or translation of Wg. Coexpression experiments ruled out a role for dSno in Notch signaling and as a partner for Brk in wing disks.Coexpression epistasis assays were able to specify where dSno might be acting in the Wg pathway Lastly, although we have not yet identified the biochemical basis for dSno - Wg pathway interactions we have continued our analysis of dSno - Smad complex formation. Our previous data showed that dSno is capable of binding to Medea and dSmad2 but not to Mad In summary, we report an unexpected developmental role for dSno as a tissue-specific protein in Wg signaling with modest antagonistic activity under normal conditions in wing development but that effectively blocks extraneous Wg signals. Genetic evidence suggests the hypothesis that dSno antagonizes Wg signaling via a protein-protein interaction mechanism in cooperation with members of the cytoplasmic Arm destruction complex. A cytoplasmic role as an antagonist of Wg signaling and a nuclear role in facilitating TGF-\u03b2 signaling may underlie the observation that the relative abundance of cytoplasmic versus nuclear SnoN is a prognostic indicator in a subset of tumors sh1402dSno, Ex17BdSno, Ex4BdSno and UAS.dSno 174dSnoGS-C517TdSnoS10Fly stocks are as described: Achaete-lacZ Ex17BdSno FRT40A or sh1402dSno FRT40A were generated by standard methods. Ex17BdSno or sh1402dSno clones in wing disks were marked with Arm-lacZ FRT40A EX17BdSno FRT40A and Arm-lacZ FRT40A were heat shocked to express FLP recombinase from an X chromosome insertion at 72\u201396 hours after egg deposition to generate numerous small clones. Arm-lacZ is ubiquitously expressed EX17BdSno or sh1402dSno by recombination were unambiguously visualized by the absence of lacZ.Mutant clones: Recombinant chromosomes carrying Gal4-UAS studies: MS1096.Gal4 is an insertion in the X-linked gene Beadex that has a hemizygous wing phenotype in males but is fully recessive in females Control experiments: Tests for Gal4 titration in strains with multiple UAS transgenes were conducted by substituting UAS.lacZ for UAS.dSno as described Statistics: To quantitate any observed lethality UAS transgenes were placed over a marked balancer in the parental strain and then the percent of expected adult progeny inheriting the transgene was calculated with reference to the number of siblings inheriting the balancer chromosome.Antibody labeling: The analysis of wing disks followed Microscopy: Images were collected on a Leica SP2 confocal microscope as a series of optical sections encompassing both cell layers of the wing disk. Each section was 0.18 \u00b5m thick and taken every 2.0 \u00b5m. Images displayed are compilations ranging in size from 14 to 24 optical sections. Images are sized to maximize visibility of the antibody labeling and are not to scale.Text S1Accompanying text, procedures and references for Supplemental Figures.(0.07 MB PDF)Click here for additional data file.Figure S1dSno mutants. (A) The coordinate line represents 105649base pairs from polytene region 28D3 (Genbank AE014134.5 - Release 5.22 sequence of D. melanogaster chromosome 2L - Dec 2009). Five resident genes (dSno is composed of two predictions CG7233 and CG7093) sized roughly to scale with their transcriptional orientations are shown above the line. The splicing pattern of the longest transcript encoding dSnoN (the longest protein isoform) is also shown. The nucleotide locations of the transcription start site and the initiator methionine for isoform are indicated below the coordinate line. (B) sh1402dSno contains a precise insertion of a P{lacW} transposon and a precise deletion (not shown) of a 297-class transposable element that is present in the 2L reference sequence. sh1402dSno is missing one of the three known dSno promoters and acts as a modest hypomorph. This data was previously shown in Ex17BdSno is a deletion of 5023 bp when compared to sh1402dSno that deletes the three known dSno promoters, the adjacent CG7231 and the 5\u2032 end of CG7228. Ex17BdSno acts as a strong hypomorph. (D) Ex4BdSno is a deletion of 20849 bp when compared to sh1402dSno that deletes all dSno promoters, CG7233 (corresponding to the dSnoI protein isoform), CG7231, CG7224, CG7228 but not CG7224. Ex4BdSno is a protein null. (E) As reported in 174dSno is a deletion of 9518 bp when compared to sh1402dSno. The deletion begins at amino acid 57 removing the remaining 276 amino acids of CG7233 and the splice acceptor creating essentially a protein null.Comparative genomic analysis of four (4.87 MB TIF)Click here for additional data file.Figure S2dSno transcription is significantly reduced in Ex17BdSno embryos and similar to Wg expression in the ventral epidermis.Embryos in lateral view. (A) Stage 17 wild type embryo hybridized with a dSnoI riboprobe displaying strong dSno expression in the brain and ventral cord. Additional expression in segmentally reiterated stripes in the ventral epidermis is indicated with red arrowheads. (B) Stage 15 homozygous Ex17BdSno embryo with weak staining in the brain and ventral cord. (C) Left side - Stage 17 transheteroygous Ex17BdSno/Ex4BdSno mutant embryo with weak staining in the brain and ventral cord that is significantly less than in wild type. Right side - Stage 17 embryo heterozygous for a dSno excision allele balanced over CyOP{wg-lacZ}. This sibling embryo is a control for embryo genotype and the staining reaction. (D)Stage 17 wild type embryo revealing that dpp RNA is present in many tissues but not in the ventral epidermis (red arrowheads). (E) Stage 16 wild type embryo with Wg protein expression visible in the ventral epidermis that corresponds to regions that will generate naked cuticle.(4.45 MB TIF)Click here for additional data file.Figure S3dSno is expressed in the optic lobe and dSno mutant optic lobes display reduced cell proliferation. A) In a wild type third instar larval optic lobe, a dSnoI riboprobe reveals prominent expression in the presumptive lamina plexus and medulla neuropil (black arrowhead). B-C) Optic lobes stained with antibodies to Brdu (green) and Elav (red). An arrowhead indicates the inner proliferation zone of the medulla neuropil. B) Wild type lobe has a well-defined inner proliferation zone containing numerous cells in S-phase. C) Transheteroygous 174dSno/Ex4BdSno mutant lobe with an ill-defined inner proliferation zone containing a reduced number of cells in S phase. This result is consistent with previous optic lobe data showing that sh1402dSno/Ex4BdSno mutants have reduced numbers of cells in M phase (1.70 MB TIF)Click here for additional data file.Figure S4Ex17BdSno wing clones and loss of function genotypes phenocopy clones of the Wg pathway antagonist zw3. Wild type wing. Wings with unmarked clones of Ex17BdSno display up to eight individual ectopic margin bristles in the distal region of the anterior compartment of the wing blade (arrowheads). Wings with unmarked clones of M11zw3 display numerous ectopic margin bristles, individual bristles as well as clusters of bristles, throughout the wing blade due to loss of Zw3 antagonism for Wg signaling. Wings with unmarked clones of the Wg transcription factor arm (4arm) are missing margin bristles due to the loss of Wg signaling. Wings of 174dSno homozygous escapers display up to ten individual ectopic margin bristles in distal and medial regions of the anterior compartment. Wings of EX4BdSno/GS-C517TdSno transheterozygous escapers display up to five ectopic margin bristles in the distal region of the anterior compartment.(7.60 MB TIF)Click here for additional data file.Figure S5dSno loss of function genotypes display ectopic sensilla, a phenotype not associated with the loss of Dpp signaling. (A) Wild type wing. (B) High magnification view of three campaniform sensilla on the dorsal surface of longitudinal vein3 (L3) in a wild type wing (arrowheads). (C) 174dSno homozygous escaper with five campaniform sensilla on L3 (four are shown - arrowheads). (D) Wing from M11zw3 has four campaniform sensilla on L3 (arrowheads). (E) Scabrous.Gal4;UAS.lacZ pupal disk stained with anti-lacZ. Note prominent expression in the L3 primordia (arrowhead). (F) Sca.Gal4; UAS.dSno wing with most of L3 missing due to antagonism of Dpp signal transduction and is also missing two of the L3 sensilla (the remaining one is indicated with an arrowhead). (G) Sca.Gal4; UAS.Mad-RNAi wing with all of L3 missing due to loss of Dpp signal transduction but all L3 sensilla are present (arrowheads). (H) Sca.Gal4; UAS.Dsh wing with ectopic bristles on L3 due to ectopic Wg signaling. (I) Sca.Gal4; UAS.Dsh, UAS.dSno rescued wing with one remaining ectopic bristle due to dSno antagonism of ectopic Wg signaling but also with most of L3 missing due to dSno antagonism of Dpp signal transduction.(7.62 MB TIF)Click here for additional data file.Figure S6dSno mutant embryos do not have altered Wg expression but they have ectopic expression of a Wg target gene in the ventral epidermis. (A) Wild type embryo. Each hemisegment (2 are shown) of the ventral cuticle contains six rows of denticles in a trapezoidal pattern pointing to the anterior and a region of equal size with no denticles. (B) en1wg homozygous loss of function embryo. All ventral cells have denticles. (C) Glawg heterozygous gain of function embryo. Tissue-specific and non-lethal wg overexpression prevents any ventral cells from producing denticles. Note that the loss of denticles is not fatal - this embryo would eventually become an adult with a Glazed eye phenotype resulting from a second round of Wg overexpression in eye disks. (D) sh1402dSno homozygous loss of function embryo. This embryo with no denticles is similar to a Gla1wg (gain of function) embryo. Note that these denticle-less embryos would eventually hatch but they do not survive past the pupal stage due to other defects. (E) Stage 13 sh1402dSno heterozygous embryo labeled to reveal the expression of segmentally reiterated stripes of Wg protein (green) and Wg RNA (red). An enhancer trap in wg present on the CyO balancer chromosome expresses lacZ and the embryo was stained with an antibody to lacZ. (F) Stage 13 homozygous sh1402dSno embryo with wild type expression of Wg protein. (G) Stage 14 wild type embryo labeled to display segmentally reiterated stripes of En expression (each En stripe is located immediately posterior to a Wg stripe and En is a target of Wg). The one to two cells wide stripe of En expression is visible in the inset. (H) Stage 14 homozygous sh1402dSno embryo with expanded En expression in each stripe. The width of each stripe of En staining is expanded to three to four cells (inset).(5.05 MB TIF)Click here for additional data file.Figure S7dSno - Medea binding is conserved between mammals and flies. (A) Deletion of amino acids 1\u201369 or 1\u2013108 from dSno did not affect Medea interaction. The T280Y mutation in dSno decreased the intensity of Medea interaction. (B) The W283E mutation in dSno abolishes Medea interaction as does the dSno double mutant T280Y and H271A. (C) Deletion of amino acids 1\u2013108 of dSno decreases recruitment of dSmad2 to dSno - Medea complexes: compare the amount of dSmad2 in lane 4 with lane 6. Reduction in dSno - Medea binding by the T280Y mutation also leads to reduced binding of dSmad2: compare lane 4 with lane 8. (D) Analysis of a deletion series covering the first 108 amino acids of dSno reveals that only the first 13 amino acids are required for dSmad2 recruitment to Medea - dSno complexes. (E) Schematic of dSno mutants with an amino acid scale bar and domains as indicated: blue is Medea interaction, purple is a coiled-coil and gray is a region of significant identity between predicted Sno proteins from 12 Drosophila species . Also shown are effects on dSno - Medea binding or Medea - dSno complex recruitment of dSmad2: + \u200a=\u200a interaction, - \u200a=\u200a no interaction, +/\u2212 \u200a=\u200a weak interaction and nd \u200a=\u200a not determined.(9.69 MB TIF)Click here for additional data file."} +{"text": "To establish the frequency of necrotizing funisitis in congenital syphilis, we conducted a prospective descriptive study of maternal syphilis in Bolivia by testing 1,559 women at delivery with rapid plasma reagin (RPR). We examined umbilical cords of 66 infants whose mothers had positive RPR and fluorescent treponemal antibody absorption tests. Histologic abnormalities were detected in 28 (42%) umbilical cords , and 38 [58%] were normal. Of 22 umbilical cords of infants from mothers without syphilis (controls), only two (9%) showed mild funisitis; the others were normal. Testing umbilical cords by using immunohistochemistry is a research tool that can establish the frequency of funisitis due to Treponema pallidum infection."} +{"text": "We have used polymerase chain reaction (PCR) to measure keratin 19 mRNA in order to detect breast cancer cells invading axillary lymph nodes. In a consecutive series of 125 patients with primary breast cancer, 75 patients had no evidence of lymph node involvement by conventional histology. A total of 530 lymph nodes from these patients were examined and 106 (20%) gave a keratin 19 product detectable by Southern hybridisation. This correlated with primary tumour size (P<0.001). These 106 nodes came from 23 patients. Thus, using this technique, 23/75 (30.6%) patients were found to have evidence of lymph node involvement who would otherwise have been designated lymph node negative."} +{"text": "Amongst 17 patients with hepatic focal nodular hyperplasia (FNH) encountered at Westmead Hospitalbetween 1981 and 1990, FNH was found in association with hepatocellular carcinoma (HCC) in three (3/17), one male and two females, one of whom also had peliosis and an hepatic adenoma. FNH was alsofound in association with other conditions which may affect hepatic function, structure or circulation,including chronic obstructive airways disease (2), congestive cardiomyopathy (1), chronic activehepatitis (1), granulomatous hepatitis (1), coeliac artery stenosis (1) and metastatic malignant melanoma(1).This report, derived from our experience with FNH over 10 years draws attention to a possible linkbetween FNH, hepatic malignancy and conditions which may disturb the hepatic circulation. We suggestthat patients with FNH should be investigated thoroughly and an aggressive management policy shouldbe adopted."} +{"text": "To the Editor:vesicle associated membrane protein associated protein B (VAPB) gene was described in eight Brazilian families of Portuguese descent showing a wide spectrum of motor neuron diseases (MNDs) including spinal muscular atrophy (SMA) and familial amyotrophic lateral sclerosis (ALS) (ALS8) (VAPB gene in a non-Brazilian patient.A dominant missense mutation p.P56S in the ) (ALS8) , 2. HaplA 43-year-old man (III-1) showed slowly progressive muscular weakness for 2 years and a family history of autosomal dominant neuromuscular disease through at least three generations . The patThe index patient (III-1) showed paresis of the hip flexors and extensors grade 4/5) and fasciculations in the proximal muscles of arms, legs on both sides. Deep tendon reflexes were normal except for absent Achilles tendon reflexes. There were no pyramidal tract signs. Needle electromyography showed fasciculations and signs of chronic denervation. Nerve conduction studies of tibial nerves revealed slightly reduced amplitudes on the left. Motor evoked potentials in both tibialis anterior muscles after magnetic stimulation of the motor cortex and the lumbar roots were normal.VAPB gene. Amplicons were purified and sequenced directly on an ABI PRISM 310 Genetic Analyzer . The p.P56S mutation was screened in 100 German healthy controls. Haplotype analysis was performed using microsatellite markers D20S100, D20S171 and D20S173 from the ABI Prism Linkage Mapping Set kit version 2 as reported previously Genomic DNA of the index case was extracted from peripheral blood and amplified using primer pairs flanking all exons and exon/intron boundaries of the VAPB gene. This mutation was not present in 200 German control chromosomes. Haplotype analysis revealed that this patient had a different haplotype compared to the Brazilian families (Sequencing revealed a heterozygous p.P56S point mutation in exon 2 of the families .VAPB mutation VAPB mutations in cohorts of patients with ALS (VAPB gene seem to be rare in familial MNDs. However, our case report demonstrates that the p.P56S mutation can be observed outside Brazil, and should be considered as a rare differential diagnosis in familial MNDs.The phenotype of our index case and his mother represented late onset SMA as observed in one-third of the Brazilian patients carrying the p.P56S with ALS \u20136]. Muta"} +{"text": "The wound healing process and production oftumour necrosis factor alpha (TNF-\u03b1) by peritonealcells of 7-day and 14-day obstructive jaundice (OJ)and sham-operated rats were investigated. In thestudy the skin wound breaking strength was measured,In addition such histological and biochemicalparameters as fibroblast and endothelial cell proliferation,inflammatory cell infiltration and hydroxyprolinecontent were evaluated in polyurethanesponge discs implanted subcutaneously into rats.TNF-\u03b1 production by peritoneal exudate cells (PEC),both spontaneous and lipopolysaccharide (LPS)-induced was determined by a bioassay. In OJ rats theprocess of both early as well as late phase of healingwas impaired. The breaking strength of skin woundwas decreased, the fibroblast and endothelial cellproliferation and collagen deposition, as well as hydroxyprolinecontent were diminished. In 7 day OJthe numbers of inflammatory cells in the implantswere lowered with a subsequent slight increase onday 14 of OJ. The spontaneous and LPS induced TNF-\u03b1 production by PEC were significantly higher in 7day OJ as compared with sham-operated controls. Onday 14 of OJ the LPS-induced TNF-\u03b1 level was, incontrast, much lower and did not differ much fromthe spontaneous TNF-\u03b1 production. We concludethat the impairment of wound healing in OJ resultsfrom disturbances in functioning of the immunesystem caused by systemic endotoxaemia."} +{"text": "However, cells derivedfrom both strains produce large amounts of IL-10 but not IL-4. Immunized lymph node cellsderived from EAE susceptible (AO \u00d7 DA) F1rats induce clinical signs of disease in sublethallyirradiated parental DA but not AO rats. The pathohistology of the target tissue in theserecipients clearly demonstrated infiltration of mononuclear cells in both parental strains.However, the number of CD4+ cells was significantly higher and number of apoptotic cellssignificantly lower in DA rats sacrificed 8 days after passive transfer. We postulate that inaddition to higher IFN-\u03b3 and TNF-\u03b1 production, resistance to early apoptosis of the invadingcells in the target tissue possibly due to lack of downregulation by TGF-\u03b2 leads to exceptionalsusceptibility to EAE in DA rats.Dark Agouti (DA) rats are highly susceptible to induction of Th-l-mediated autoimmunitydisease, including experimental allergic encephalomyelitis (EAE). In contrast to other susceptiblerat strains in which disease is induced only with encephalitogen emulsified in completeFreund's adjuvants (CFA), in DA rats EAE develops after injection of encephalitogen inincomplete Freund's adjuvants (IFA) or Titermax, putative Th-2 directed adjuvant. Lymphnode cells derived from immunized DA rats and stimulated"} +{"text": "Data were collected on subsequent primary cancers occurring in 194 individuals diagnosed with ampullary carcinoma during 1979-92 in the north western region of England, UK. Four cancers were identified compared with 6.62 expected (relative risk 0.60), suggesting that individuals with ampullary carcinoma are not at increased risk of developing subsequent primary cancers."} +{"text": "Extrapolation to humans from experimental radioimmunotherapy in nude mouse xenograft models is confounded by large relative tumour size and small volume of distribution in mice allowing tumour uptake of radiolabelled antibodies unattainable in patients. Our large animal model of human tumours in cyclosporin-immunosuppressed sheep demonstrated tumour uptake of targeted radiolabelled monoclonal antibodies comparable with uptakes reported in clinical trials. Sheep immunosuppression with daily intravenous cyclosporin augmented by oral ketoconazole maintained trough blood levels of cyclosporin within the range 1000-1500 ng ml(-1). Human tumour cells were transplanted orthotopically by inoculation of 10(7) cells: SKMEL melanoma subcutaneously; LS174T and HT29 colon carcinoma into bowel, peritoneum and liver; and JAM ovarian carcinoma into ovary and peritoneum. Tumour xenografts grew at all sites within 3 weeks of inoculation, preserving characteristic morphology without evidence of necrosis or host rejection. Lymphatic metastasis was demonstrated in regional nodes draining xenografts of melanoma and ovarian carcinoma. Colonic LS1 74T xenografts produced mucin and carcinoembryonic antigen (CEA). The anti-CEA IgG1 monoclonal antibody A5B7 was radiolabelled with iodine-131 and administered intravenously to sheep. Peak uptake at 5 days in orthotopic human tumour transplants in gut was 0.027% DI g(-1) (percentage of injected dose per gram) and 0.034% DI g(-1) in hepatic metastases with tumour to blood ratios of 2-2.5. Non-specific tumour uptake in melanoma was 0.003% DI g(-1). Uptake of radiolabelled monoclonal antibody in human tumours in our large animal model is comparable with that observed in patients and may be more realistic than nude mice xenografts for prediction of clinical efficacy of radioimmunotherapy."} +{"text": "Vascular endothelial growth factor (VEGF) expression, vascularisation and tumour cell proliferation were analysed in 91 human epidermoid lung carcinomas using immunohistochemistry. A polyclonal anti-VEGF antibody was used for VEGF expression, a polyclonal antibody directed against human von Willebrand factor (factor VIII) to identify blood vessels and the proliferating cell nuclear antigen (PCNA) as a marker for proliferating cells. Positive staining for VEGF was obtained in 54 out of 91 cases (59%), the number of blood vessels varied from zero to 64 counts (mean 9.4) and the proportion of PCNA-positive cells varied from 1.3% to 72.1% (mean 25.2%). The mean PCNA labelling index and mean microvessel count in VEGF-positive tumours were significantly higher than those in VEGF-negative tumours . In addition, PCNA labelling index significantly increased with increasing VEGF expression . In contrast, no association was found between PCNA labelling index and tumour vascularity . The close correlation of VEGF expression with tumour cell proliferation and microvessel density suggests that VEGF acts both as an autocrine growth factor and as stimulator for angiogenesis. However, tumour cell proliferation and microvessel growth and/or density may be regulated by separate mechanisms."} +{"text": "Lancet343: 86\u201387; McDonald et al (1995) Cancer Epidemiol Biomarkers Prevent4: 791\u2013793). Of the 29 archived samples of SCC meeting quality criteria for DNA analysis by polymerase chain reaction (PCR) and Hae III restriction enzyme digestion, two tumours were found that harboured this mutation. DNA sequencing confirmed the presence of a G to T base substitution within the Hae III site spanning codons 249 and 250 of the p53 gene that results in replacement of arginine (wild-type) by methionine at residue 249. When these data are combined with those from our previous study of tumours from the Stollberg Archive in which 50 lung tumours were examined, (including nine SCCs), we conclude that the G\u2192T (arg\u2192met) codon 249 mutation prevalence in the Wismut miner cohort is not sharply elevated in lung cancers in general (two mutations/79 tumours), or specifically in SCCs of the lung (two mutations/38 SCC) when compared to data from lung cancer patients with no reported occupational exposure to radon gas. \u00a9 2000 Cancer Research CampaignSquamous cell lung carcinomas (SCC) from former employees of the Wismut uranium mining company were obtained from the Stollberg Archive in order to screen for p53 tumour suppressor gene codon 249 arg\u2192met hotspot mutations, a putative molecular bio-dosimeter of alpha-particle (radon) exposure"} +{"text": "Using flow cytometric DNA analysis of paraffin embedded tissue, DNA histograms were successfully obtained from the anal cancers of 117 patients. DNA diploid patterns were given by 82 cancers (70%) and DNA non-diploid patterns by 35 cancers (30%): 15 DNA aneuploid, 20 DNA tetraploid. Well differentiated squamous cell cancers were mainly DNA diploid, while a larger proportion of poorly differentiated and small cell cancers were DNA non-diploid. The large majority of stage A cancers were DNA diploid. A greater proportion of tumours that had invaded through the anal sphincter or had lymph node metastases or distant spread were DNA non-diploid. Prognosis was slightly poorer for patients with DNA non-diploid cancers when compared to patients with DNA diploid tumours (P = 0.08) and significantly poorer for individuals with DNA aneuploid anal cancers (P = 0.037). However, in a multivariate analysis model, the DNA ploidy pattern of an anal cancer was not of independent prognostic significance alongside tumour histology and tumour stage."} +{"text": "The mean electrophoretic mobility (EPM) of splenic cells was determined in 4 different host-tumour systems. In splenic cells harvested from mice bearing slow-growing tumours, a significant EPM decrease was observed (12%) and in increase in the proportion of cells with slow mobility. Moreover, after 1 h incubation in RPMI medium at 37 degrees C and 2 washings, spleen cells showed a marked increase in their EPM (average 30%). Finally, the supernatant from incubation medium after contact with normal spleen cells (1 h at 37 degrees C) produced a significant decrease in their EPM (approximately 12%). On the other hand, no significant EPM variations were found between control spleen cells and cells from fast-growing tumours, before or after 1h incubation. The existence of factors which induce a loss of negative surface charge on spleen cells of some tumour-grafted mice is discussed."} +{"text": "Of 297 patients with metastatic testicular and extragonadal germ cell tumours (GCT), bone involvement was detected clinically in 3% (7/251) of those at first presentation and in 9% (4/46) of relapsed cases. This difference was not statistically significant . Concurrent systemic metastases, commonly involving lung (7/11 cases) and para-aortic lymph nodes (6/11), were present in all patients with bone disease. All affected patients had localized bone pain and lumbar spine was the most frequent site involved (9/11). Spinal cord compression occurred in two patients while a third developed progressive vertebral collapse after chemotherapy and required extensive surgical reconstruction. At median follow-up of 4 years, survival among patients presenting with bone disease (6/7) was similar to overall survival in the whole group (84%) and appeared better than in those with liver or central nervous system (6/9) metastases at presentation. Back pain in metastatic germ cell tumours is often due to retroperitoneal lymphadenopathy but lumbar spine osseus metastases must be recognized early if severe potential complications, such as spinal cord compression, are to be avoided. In this series, bone metastases were not seen in the absence of widespread systemic disease suggesting all solitary bony lesions in GCT patients should be biopsied."} +{"text": "Continuous hepatic artery floxuridine infusion benefits patients with colorectal liver metastases. Implanted infusion pumps are more expensive but may result in fewer treatment interruptions than when using an external pump connected to a port. We have assessed device-related complications, treatment interruptions and added nurse interventions in 95 patients undergoing a total of 959 treatment cycles via either implanted pump (64 patients) or port (31 patients). Compared with the implanted pump, the port was associated with a significant increase (P < 0.003) in catheter blockage (24/31 vs 2/64 patients), treatment interruption (15/265 vs 12/694 treatments) and added nurse intervention (80/265 vs 20/694 treatments). Survival in patients with colorectal liver metastases is limited and the complications of treatment should be kept to a minimum. An implanted subcutaneous infusion pump offers the benefit of a 3-fold lower incidence of treatment interruption and a 30-fold lower incidence of catheter blockage than when continuous infusion chemotherapy is given via an external infusion device."} +{"text": "Although micronucleus frequency at 2 Gy differed widely between tumours evaluated , no significant correlation was observed between this parameter and clinical outcome. The average increment of micronucleus frequency after 20 Gy amounted to 193% of spontaneous level (range 60\u2013610%) and was independent of spontaneous micronucleation before radiotherapy. In contrast to in vitro results, these from in vivo assay seem to have a predictive value for radiotherapy of cervix cancer. The micronucleus increment in vivo that reached at least 117.5% of pretreatment value (first quartile for MN20 data set) correlated significantly with better tumour local control (P < 0.008) and overall survival (P < 0.045). Our results suggest that evaluation of increment of micronucleus frequency during radiotherapy (after fixed tested dose of 20 Gy) offers a potentially valuable approach to predicting individual radioresponsiveness and may be helpful for individualization of treatment strategy in advanced stage cervical cancer. \u00a9 1999 Cancer Research CampaignA potential usefulness of micronucleus assay for prediction of tumour radiosensitivity has been tested in 64 patients with advanced stage (II B\u2013IV B) cervical carcinoma treated by radiotherapy. The study of cellular radiosensitivity in vitro was conducted in parallel with the study of cellular damage after tumour irradiation in vivo. Radiosensitivity of in vitro cultured primary cells isolated from tumour biopsies taken before radiotherapy was evaluated using cytokinesis-block micronucleus assay. Frequency of micronuclei per binucleated cell (MN/BNC) at 2 Gy was used as a measure of radiosensitivity. Radiation sensitivity in vivo was expressed as per cent increment of micronucleus frequency in cells isolated from biopsy taken after 20 Gy over the pre-treatment spontaneous micronucleus level and was called MN20. Very low correlation ("} +{"text": "Four hundred and fifteen males and 367 females who had invasive malignant tumours of the salivary glands as their first cancer diagnosed in Connecticut between 1935 and 1978 were identified and followed 2342 and 2868 person-years respectively. Overall a slight excess of second primary cancers (relative risk 1.35) was observed. Significant excesses were noted for respiratory cancers in males (relative risk 2.8) and for ovarian cancer (relative risk 5.3) but not breast cancer (relative risk 1.3) in women. Possible reasons for excesses at these sites are discussed, but it seems most likely they are related to small number variation."} +{"text": "Antiphospholipid antibodies (aPL) have been found in the blood of patients with systemic andneurological disease. The rare reports of aPL in cerebral spinal fluid (CSF) have been limited mostly toIgG and IgM anticardiolipin (aCL). Our published finding of IgA aPE in the CSF of a young strokevictim prompted us to establish \u201cnormal\u201d CSF aPL values for a panel of aPL, which included aCL,antiphosphatidylserine (aPS), antiphosphatidylethanolamine (aPE) and antiphosphatidylcholine (aPC).CSF samples were tested by ELISA for IgG, IgM and IgA aPL. In addition, the CSF samples weretested for activity in the presence and absence of phospholipid (PL) binding plasma-proteins. A total of24 data points were obtained for each CSF sample.We tested 59 CSF samples obtained from 59 patientswho were undergoing evaluation for systemic or neurologic diseases. All CSF samples had normalprotein, glucose and cell counts. Ten of the 59 CSF samples (17%) had elevated aPL optical density(OD) values an order of magnitude higher than the other 49 CSF samples for one or more aPLspecificity and/or isotype. One CSF sample had both PL-binding protein dependent and independentIgG aPE activity. Another CSF sample showed both IgG aPE and aPC reactivity. The remaining eightCSF samples showed single aPL findings; IgG aPE (5), IgG aPC (1), IgG aCL (1) and IgM aPC (1).Seven of 10 patients with elevated CSF values were females. As expected, most \u201cnormal\u201d aPL ODvalues were substantially lower in CSF than those we have reported in blood samples from volunteerblood donors."} +{"text": "Five patients with hepatic (3), pelvic (1) or spinal (1) hydatid cysts received 10 mg/kg/d albendazole for 1\u20133months prior to surgery. Daughter cysts were present in the spinal hydatid and in one patient with hepaticdisease. Electron microscope examination of the cyst tissue of the pelvic and the 2 hepatic cysts lackingdaughter cysts showed no evidence of germinal layer, and the protoscoleces were dead. The primary cyst ofthe hepatic hydatid with daughter cysts (1 month therapy) was also judged dead but some pieces of thedaughter cyst germinal layer appeared normal and had unaffected protoscoleces. The daughter cyst tissue ofthe spinal hydatid (3 month therapy) appeared normal and the protoscoleces viable. In view of theundetermined viability of human hydatids before chemotherapy, treatment of longer than 1 month isadvocated for hepatic cysts, particularly if daughter cysts are present, and longer therapy is indicated forspinal disease."} +{"text": "A perifusion technique for microscopy with computerized detection of early changes in cell morphology during continuous perifusion was used to show that the geometry of cultured glioma cells (MG-251) changes rapidly when they are exposed to estramustine phosphate (EMP). When the cells were exposed to 20 or 40 mg l(-1) EMP, cell volume projected cell area (PCA) rapidly increased. When the Na+,K+-ATPase blocker ouabain (100 micromol l(-1)) was added to the EMP (40 mg l(-1)) perifusion, the acute EMP response was eradicated. When the PCA curve for ouabain alone was subtracted from the curve of combined ouabain and EMP perifusion, the resulting curve showed that ouabain completely blocked the EMP-induced increase in PCA. When the Na+, K+, Cl- co-transport inhibitors bumetanide (10 micromol l(-1)), or furosemide (100 micromol l(-1)), were added to EMP (40 mg l(-1)), the acute increase in PCA seen for EMP alone was also completely blocked. This study shows that inhibitors of ion transmembrane transport can modify EMP-induced cell volume increases. This may be of particular importance since the blockers have been found to interfere also with the cytotoxic function of EMP during cell culture. Thus, it is possible that cell volume changes could serve as a rapid technique for predicting the cytotoxic activity of antineoplastic drugs."} +{"text": "Although restricted transhepatic portal flow is necessary for development of generalized portalhypertension (GPH), increased splanchnic arterial inflow also contributes to GPH and its clinicalsequelae. In this context, we describe 7 male and 6 female patients (mean age 48 years) in whom thelesser splanchnic (gastrosplenic) system played a key role in the signs and symptoms of GPH. These 13patients shared common features of massive splenomegaly, huge splenofundic gastric varices, oftenwith a prominent natural shunt to the left renal vein. Total or near total splenectomy alone or combinedwhere appropriate with coronary vein ligation was effective in controlling varix hemorrhage (10patients), ascites (3), or complications of an enlarged spleen-anorexia and abdominal pain (3),hemolytic anemia (1) and profound thrombocytopenia with severe epistaxis (1). Intraoperative jejunalportal venography was crucial in operative management in order to establish definitively the presence orabsence of coronary venous collaterals, and when present, to verify their operative ligation.\t\tThese distinctive patients illustrate: 1) GPH is a heterogeneous syndrome of divergent splanchniccirculatory patterns, a feature which should be taken into account in selecting operative treatment; 2)one well-defined subgroup displays prominent hyperdynamic lesser splanchnic and specifically, splenicblood flow as a major contributor to clinical complications; and 3) within this subgroup, splenectomycombined with documented absence or surgical interruption of coronary venous collaterals as corroboratedby intraoperative portography is effective alternative treatment."} +{"text": "We retrospectively analysed neuromuscular toxicity associated with paclitaxel 210 mg m(-2) given by 3-h infusion in 247 patients. The severity correlated significantly with total cumulative dose, but could not be predicted by the pretreatment clinical variables or by pharmacokinetic parameters. The toxicity tended to occur in early treatment cycles."} +{"text": "Circulating tumour cells play a central role in the metastatic process, but little is known about the relationship between this cellular subpopulation and the development of secondary disease. This study was aimed at assessing the presence of colonic cells in peripheral blood of patients with colorectal cancer in different evolutionary stages, by means of reverse transcriptase polymerase chain reaction (RT-PCR) targeted to carcinoembryonic antigen (CEA) mRNA. In vitro sensitivity was established in a recovery experiment by preparing serial colorectal cancer cell dilutions. Thereafter, 95 colorectal cancer patients and a control group including healthy subjects (n=11), patients with other gastrointestinal neoplasms (n=11) or inflammatory bowel disease (n=9) were analysed. Specific cDNA primers for CEA transcripts were used to apply RT-PCR to peripheral blood samples. Tumour cells were detected down to five cells per 10 ml blood, thus indicating a sensitivity limit of approximately one tumour cell per 10(7) white blood cells. CEA mRNA expression was detected in 39 out of 95 colorectal cancer patients (41.1%), there being a significant correlation with the presence of distant metastases at inclusion. None of the healthy volunteers and only 1 of 11 patients (9.1%) with other gastrointestinal neoplasms had detectable CEA mRNA in peripheral blood. By contrast, CEA mRNA was detected in five of the nine patients (55.6%) with inflammatory bowel disease. These results confirm that it is feasible to amplify CEA mRNA in the peripheral blood, its presence being almost certainly derived from circulating malignant cells in colorectal cancer patients. However, CEA mRNA detectable in blood of patients with inflammatory bowel disease suggests the presence of circulating non-neoplastic colonic epithelial cells."} +{"text": "Chronic hepatitis C virus (HCV) infection is a leading cause of end-stage liver disease worldwide and HCV genotype 4 (HCV4) is predominant in African and Middle Eastern countries. It is well established that interferon-\u03b1 (IFNa) treatment for HCV may trigger serum autoantibodies against pancreatic islet cells (ICA) in a subgroup of patients. Available data on the incidence of ICA during IFNa therapy for chronic HCV4 infection are not conclusive. We investigated the appearance of ICA in 40 na\u00efve Egyptian patients with histologically defined chronic HCV4 infection undergoing IFNa treatment at a dose of 9-million U/week for 24 weeks. Serum samples were collected at baseline and following IFNa therapy and ICA were detected using indirect immunofluorescence. Baseline evaluation indicated that 2/40 (5%) patients had detectable serum ICA. After the completion of the treatment scheme, 12/38 (32%) previously ICA negative patients became ICA positive; however, no patient developed impaired glucose tolerance (IGT) or diabetes during follow-up. In conclusion, we submit that IFNa treatment for chronic hepatitis C (CHC) may induce serum ICA in one-third of Egyptian patients with HCV4. These autoantibodies, however, do not lead to alterations in glucose metabolism."} +{"text": "Metastatic osteosarcoma (OS) has a very poor prognosis. New treatments are therefore wanted. The conditionally replicative adenovirus Ad5-\u039424RGD has shown promising anti-tumor effects on local cancers, including OS. The purpose of this study was to determine whether intravenous administration of Ad5-\u039424RGD could suppress growth of human OS lung metastases. Mice bearing SaOs-lm7 OS lung metastases were treated with Ad5-\u039424RGD at weeks 1, 2 and 3 or weeks 5, 6 and 7 after tumor cell injection. Virus treatment at weeks 1\u20133 did not cause a statistically significant effect on lung weight and total body weight. However, the number of macroscopic lung tumor nodules was reduced from a median of >158 in PBS-treated control mice to 58 in Ad5-\u039424RGD-treated mice (p = 0.15). Moreover, mice treated at weeks 5\u20137 showed a significantly reduced lung weight , a significantly increased body weight gain and a reduced number of macroscopic lung tumor nodules compared to PBS treated control animals. Adenovirus hexon expression was detected in lung tumor nodules at sacrifice three weeks after the last intravenous adenovirus administration, suggesting ongoing viral infection. These findings suggest that systemic administration of Ad5-\u039424RGD might be a promising new treatment strategy for metastatic osteosarcoma. Osteosarcoma is the most prevalent non-hematological primary malignant bone tumor. They can be subdivided by location (medullary and surface) and by grade (high or low grade). The vast majority of high-grade osteosarcoma comprises of conventional osteosarcoma, but also includes high-grade surface, telangiectatic and small-cell osteosarcoma . PatientWe explore the use of conditionally replicative adenoviruses (CRAds) as a potential new treatment modality for high grade OS and its lung metastases. Virus replication will lyse tumor cells and released viral progeny can infect neighboring tumor cells leading to lateral spread and increased tumor cell kill. Previously we found that primary OS cells express low levels of the high affinity receptor for adenoviruses, the coxsackie and adenovirus receptor (CAR) ,11. In cLocal treatment of OS with CRAds has shown promising results in animal models ,18. Howe9 plaque forming units (pfu) intravenously once a week for three consecutive weeks . This was the practically highest achievable dose and injection of this amount of virus did not affect body weight, suggesting that it did not induce overt toxicity. Therefore, this dose was chosen to explore the anticancer potency against OS lung metastases. SaOs-lm7 cells were injected intravenously into mice to establish OS lung metastases [9 pfu Ad5-\u039424RGD or with PBS at weeks 1, 2 and 3 after SaOs-lm7 injection. As parameter of mouse well-being, their relative gain in body weight was calculated. Ten weeks after tumor cell injection, mice were sacrificed and their lungs dissected. Mouse lung weight and number of macroscopic lung tumor nodules were quantified to assess Ad5-\u039424RGD anticancer effect. Mice treated with Ad5-\u039424RGD or PBS showed no significant difference in body weight gain during the experiment (data not shown). As shown in figure We first performed a pilot study in which mice were given 1 \u00d7 10tastases . In a fiIn a next experiment, we delayed the onset of oncolytic adenovirus treatment by 4 weeks. Virus or PBS was injected once per week at 5, 6 and 7 weeks after SaOs-lm7 tumor cell injection. Mice treated with Ad5-\u039424RGD showed an average increase in weight of 18%, which was significantly higher than the average 8% weight gain observed for PBS treated control animals (P < 0.005). Moreover, intravenous administration of Ad5-\u039424RGD resulted in a significant reduction of lung weight (median 241 mg) compared to PBS treated control animals . Mice were injected with 5 \u00d7 106 SaOs-lm7 cells in the lateral tail vein in 200 \u03bcl Hanks Balanced Salt Solution w/o calcium and magnesium. Intravenous injection of SaOs-lm7 cells results in microscopic lung metastases by week 3\u20135 and macroscopic disease at 6 weeks.Female athymic/nude/nude mice were purchased from Harlan . Animals were maintained under pathogen-free conditions and fed a standard laboratory diet 9 pfu Ad5-\u039424RGD in 200 \u03bcl PBS and the second control group received 200 \u03bcl PBS.In two independent experiments, 12 mice bearing SaOs-lm7 lung metastases were randomly divided into 2 groups of each 6 mice. Mice were treated three times, in weeks 1, 2, and 3 (experiment 1) or in weeks 5, 6 and 7 (experiment 2). The first group received intravenous injections in the tail vein of 1 \u00d7 10Body weight was measured twice a week. All mice were sacrificed in week 10 after tumor cell injection. The lungs were removed in toto without difficulty and photographed. Two researchers counted tumor nodules on the outside of the lungs independently and the average of the two measurements was calculated. Lungs were snap frozen in liquid nitrogen and weighed using a Mettler H31 .Cryosections of lungs obtained at week 10 from mice treated at weeks 5, 6 and 7 after tumor cell injection were fixed in acetone. Immunohistochemical staining for adenovirus (hexon) was done using goat anti-adenovirus antibody 1056 . Sections were then incubated with biotinylated goat anti-rabbit IgG antibodies. The immune reaction was visualized using the streptavidin-biotin-Horse Radish peroxidase complex method followed by diaminobenzidine solution . Sections were counterstained with hematoxylin , dehydrated and mounted.Differences in relative gain in mouse weight, number of lung metastases and total lung weight between treatment groups were analyzed with a two and one-tailed Mann-Whitney test, respectively, using GraphPad Instat 3.0 .The author(s) declare they have no competing interests.HCAG: designed and carried out the experiments and drafted the manuscriptVWB: designed experiments, analyzed data and critically commented on final manuscriptFHES: counted OS lung tumor metastases and corrected manuscriptMAW: optimized mouse model and corrected manuscriptESK: optimization of mouse model, and commented on manuscriptMNH: designed experiments and corrected manuscriptWRG: supervised project and commented on manuscriptGJLK: obtained funding, supervised project and corrected final manuscriptPIJMW: obtained funding, supervised project and corrected final manucriptAll authors read and approved the final manuscript."} +{"text": "Two rapidly growing allogeneic tumours, sublines of Yoshida (Y-P388) and Walker (W-256) injected intravenously in single cell suspensions produced tumour macrocolonies in the lungs of rats within 7 days. Y-P388 produced similar but fewer colonies in the kidneys. Colony forming efficiency (CFE) in lung was high in weanling rats given either sublethal whole body irradiation (WBI) or a single dose of rabbit anti-rat lymphocytic serum (ALS) to suppress immunity. In immunologically intact weanlings CFE was much lower and many 7-day old colonies showed signs of regression. CFE for primary tumour cell challenges decreased rapidly and markedly with increase in age of host during the first 1-2 weeks after weaning. This resistance to growth of a primary challenge in lungs of older rats was not significantly reduced by WBI but was decreased by ALS. CFE of a secondary challenge of tumour cells injected intravenously in rats which had been previously immunized with heavily irradiated (HR) tumour cells was very low; it was not significantly increased by WBI but was moderately increased by ALS. In weanling rats given lethal (900 rad) WBI, 1 hour before intravenous injection of tumour cells, treatment with bone marrow (BM) cells derived from normal adult donors increased CFE, whereas BM (or spleen) cells from immunized donors decreased CFE. The results suggest that ALS and WBI not only increase tumour CFE by suppressing immunity to tumour growth but also \u201ccondition\u201d host tissue (tumour bed) in such a way as to facilitate the survival, \u201ctake\u201d and initial replication of grafted tumour cells before the rats recover from the immunosuppressive effects of these treatments."} +{"text": "The efficacy of high-dose chemotherapy (HDC) and circulating progenitor cell (CPC) transplantation in metastatic breast cancer (MBC) relies mainly on giving this treatment after a response to conventional induction chemotherapy has been achieved. For this reason an optimal mobilization regimen should be therapeutically effective while minimizing the number of leucaphereses required to support the myeloablative therapy. The combination of an anthracycline and paclitaxel in chemotherapy-untreated MBC has produced impressive response rates. We evaluated the CPC-mobilizing capacity of the combination epirubicin (90 mg m(-2)) and paclitaxel (135 mg m(-2)) followed by filgrastim (5 microg kg(-1) day(-1)) starting 48 h after chemotherapy administration in ten patients with MBC who were eligible for an HDC and CPC transplantation programme. Leucaphereses were performed by processing at least two blood volumes per procedure at recovery from neutrophil nadir when CD34+ cells in the peripheral blood exceeded 20 microl(-1). In most patients (six out of 10) more than 2.5 x 10(6) CD34+ cells kg(-1), a threshold considered to be sufficient for haematopoietic reconstitution, were collected with a single apheresis. In the remaining four patients an additional procedure, performed the following day, was enough to reach the required number of progenitors. These data suggest that the epirubicin-paclitaxel combination, besides being a very active regimen in MBC, is effective in releasing large amounts of progenitor cells into circulation."} +{"text": "The genetic closeness and divergent muscle growth rates of broilers and layers make them great models for myogenesis study. In order to discover the molecular mechanisms determining the divergent muscle growth rates and muscle mass control in different chicken lines, we systematically identified differentially expressed genes between broiler and layer skeletal muscle cells during different developmental stages by microarray hybridization experiment.Taken together, 543 differentially expressed genes were identified between broilers and layers across different developmental stages. We found that differential regulation of slow-type muscle gene expression, satellite cell proliferation and differentiation, protein degradation rate and genes in some metabolic pathways could give great contributions to the divergent muscle growth rates of the two chicken lines. Interestingly, the expression profiles of a few differentially expressed genes were positively or negatively correlated with the growth rates of broilers and layers, indicating that those genes may function in regulating muscle growth during development.The multiple muscle cell growth regulatory processes identified by our study implied that complicated molecular networks involved in the regulation of chicken muscle growth. These findings will not only offer genetic information for identifying candidate genes for chicken breeding, but also provide new clues for deciphering mechanisms underlining muscle development in vertebrates. Myogenesis is a very complicated but precisely regulated developmental process whose mal-regulation can cause many neuronal, heart and muscular dysfunction diseases. During the embryonic period, muscle development starts with the emergence of somites. Myogenic precursor cells originated from somites first give rise to myoblasts, which will further undergo proliferation, migrate to their final locations and fuse into multinucleated myotubes, finally differentiate into mature muscle fibres . In geneet al. found the hypothalamic somatostatin (SS) mRNA expression was higher in layers than in broilers whereas the expression of hepatic GH receptor (GHR) mRNA was opposite [et al. also reported that broilers were less sensitive or responsive to peripheral concentrations of leptin than layers [Many researchers have made great efforts to investigate the divergent muscle growth rates and muscle cell sizes of broilers and layers and attempted to decipher the underlining mechanisms of above phenomena. Some previously published works observed that broilers have higher food intake , less acopposite . Cassy Sn layers . Severaln layers ,18. A fen layers ,20. At tn layers -23.Despite these progresses, genome-wide systematic study for the genetic basis and molecular mechanisms implicated in the divergent muscle growth rates between broilers and layers is still lacking. In the present study, we used Affymetrix chicken genome array to systematically identify differentially expressed genes in skeletal muscle tissues between broilers and layers at five posthatch developmental stages. A total of 543 differentially expressed genes were identified between broilers and layers. Functional analysis showed that muscle development related GO terms were enriched among differentially expressed genes. Greater expression ratio of type II to type I fiber genes and more activity of genes related to satellite cell proliferation and differentiation were observed in broilers than in layers, which could be important factors contributing to the divergent muscle growth rates of the two chicken lines. We also identified a few genes whose expression profiles were highly correlated with the growth rates of broilers and layers, and found several metabolic pathways significantly over-presented among differentially expressed genes in broilers and layers.2 > 99.8%), indicating that our data contained very limited artificial variance.Broiler and layer chickens showed significantly divergent skeletal muscle growth rates during development, and the body weight increment difference between broilers and layers is most significant during posthatch 2\u20136 weeks with changes \u2265 1.5 fold and p-value < 0.01 between broilers and layers considering all examined time points. The estimated False Discovery Rate (FDR) of DE genes was 0.02%. Among the 543 genes, 366 are known genes and the rest 177 are unknown genes, including ESTs and hypothetical ORFs was performed on 52 randomly selected DE genes. Among them, 46 (81%) genes exhibited expression patterns consistent with the microarray results , myoglobin (MB), myosin light chain 3 (MYL3), myosin heavy chain 7B (MYH7B) and myosin L2B regulatory light chain (MYL2B), were significantly higher in layers than in broilers , intestinal 15 kDa protein (FABP6) homolog, and elongation of long chain fatty acids (ELOVL family member 6). Genetic variation in bovine FABP4 gene has been found to be associated with intramuscular fat content and subcutaneous fat depth [FABP4 and FABP6 were higher in broilers than in layers, in agreement with more intramuscular fat deposition of broilers.Some glucose synthesis accelerating enzyme encoding genes, at depth . In our PDK4) and 3-oxoacid CoA transferase 1 (OXCT1), which play important regulatory roles in glucose and fatty acid metabolic networks [COMT) was another metabolic related gene. It has been shown that inhibition of COMT expression resulted in body weight lose in human [COMT in layers than in broilers. Notably, the Tyrosine metabolism and synthesis pathway and degradation of ketone bodies pathway were found significantly enriched of differentially expressed genes is a key enzyme for testosterone synthesis. Testosterone has pronounced effects on muscle protein synthesis and muscle mass enlargement, especially during the period of rapid muscle cell growth [HSD17B7 was observed in broilers, which might result in higher testosterone level thereby contributed to fast muscle growth and hypertrophy.Other two interesting differentially expressed genes were pyruvate dehydrogenase kinase 4 , as well as several other E3 ligases . Such overall lower expression of ubiquitin related protein degradation genes in broilers than in layers might reflect the lower degradation rate of skeletal muscle proteins in broilers, thereby contributing to the larger body sizes of broilers.Ubiquitin mediated proteolysis regulates protein abundance and serves as a central regulatory function in many biological processes. It has been shown that the protein degradation rate in broilers is lower than in layers during muscle development ,18. HoweBecause the growth rate of skeletal muscle cells vary a lot during development between broilers and layers, we wanted to identify genes with expression traits significantly correlated with chicken muscle cell growth rate, these genes might directly or indirectly involve in muscle cell growth control. Because the skeletal muscle tissue contributes the majority body weight of chicken , we usedWe also calculated the FDRs of QTE results by similar random permutation methods used for DE genes. As a result, the FDRs were 4.4%, 4.8% and 0.18% for identified QTE genes in broilers, layers and common genes between them, respectively.DKK3) gene could be a potential QTE gene both in broilers and layers. DKK3 is a tumour suppressor which could inhibit proliferation of many cancer cells [DKK3 in muscle development regulation, we found it expressed at a relative high level in skeletal muscle cells of both chicken lines. In addition, significantly higher expression level of DDK3 in broilers than in layers was observed were accumulated in broilers than in layers. In general, type-II fibres have larger diameter and are easier to response to various stress induced muscle hypertrophy. Accumulation of type-II fibres might be a major contributor to the fast growth rate of broilers at 4 weeks. Table .FHL2 and CSRP3 [MUSTN1 [The postnatal muscle growth was mainly contributed by muscle fibre hypertrophy resulting from satellite cell activation, proliferation, differentiation and fusion into the existing fibres. We found that the differential regulation of satellite cell activities might account a lot for divergent muscle growth of broilers and layers. Several growth factors and growth related genes, which play pivotal roles in regulating muscle growth and hypertrophy as strong stimulators or inhibitors of myoblast and satellite cell proliferation and differentiation, were differentially expressed between the two chicken breeds. In addition, we also identified several differentially expressed potential regulators for satellite cell proliferation and differentiation, including LIM-domain containing protein encoding genes nd CSRP3 ,28, as w [MUSTN1 .PDHX, ALDH1A2, AER61, AOX1) and fatty acid transportation and utilization related proteins (FABP4 and FABP6) expressed higher in broilers than in layers, whereas a fat break-down related enzyme thiolesterase B expressed higher in layers, suggested that different metabolic regulatory networks are indispensable for the differential growth rates between broilers and layers. The expression of 3-oxoacid CoA transferase 1 (OXCT1), a key enzyme of ketone body utilization, was higher in broilers than in layers. Skeletal muscle is one of the main target organs for ketone utilization. Previously published work showed that OXCT1 expression was increased during anaerobisis [et al found that OXCT1 is one of the most abundantly expressed proteins in human hepatocarcinoma cell line SMMC-7721 [The microarray data also helped us to investigate the mechanisms of metabolic rate difference giving rise to divergent muscle growth and hypertrophy between broilers and layers. In our microarray data, many broiler and layer differentially expressed genes were classified into functional categories involved in metabolic processes. Genes encoding some glucose metabolic related enzymes and egg-type layers (White Leghorn) were obtained from China Agricultural University. All chickens were grown under conditions in accordance with the University's Animal Care and Use Committee policy. All chickens were kept in floor pens under the same temperature and lighting conditions. For the first two days after hatch, chickens were kept in floor pens at 35\u00b0C under constant light. From the post-hatching third day to 8 weeks, temperature was reduced from 35\u00b0C to 18\u00b0C in a stepwise manner and the lighting schedule was maintained at a 18 h light and 6 h darkness cycle. Both chicken lines were fed with the same commercial layer diet. All chickens were weighed weekly. At each time point , 5 randomly selected male individuals from each group were sacrificed for tissue sampling. \u03bcg/\u03bcl. Affymetrix Gene Chip microarray hybridization was carried out according to the Affymetrix Expression Analysis Technical Manual by GeneTech Biotechnology Limited Company . Array scanning and data extraction were carried out following the standard protocol.Total RNA was isolated from breast muscle cells using Trizol reagent (Invitrogen) according to the manufacturer's instructions. Twenty microgram total RNA was suspended in RNase-Free water with a final concentration of 1.5 \u00ae) of our microarray data were detected using Affymetrix MAS5 method implemented in Bioconductor packages. Present probe-sets were defined as present in samples of at least one examined time point and were used for all following studies. The expression value of each probe set was normalized and calibrated using the RMA method. Differentially expressed probe-sets between broilers and layers were identified by cutoff of fold-change \u2265 1.5 and p-value < 0.01 in ANOVA test. To assess the False Discovery Rate (FDR) of the DE gene result, a control expression sets were generated by randomly permuting the expression values of all arrays for each gene 500 times. As a result, the estimated FDR of our approach for detecting DE genes was 0.02%; such high specificity would mainly result from the relative large sample size (N = 30).The present genes is the body mass, W0 is the initial body mass immediately after hatch (at time t0), r is the absolute growth rate, both r are unknown parameters. The average initial body mass of each chicken line was used as W0, r were calculated independently for broilers and layers by Gauss-Newton nonlinear least square regression. Then the absolute Growth Rate (GR) and the Natural Increase Rate (NIR) of chicken body mass could be calculated bywhere iBy the later function, the NIR of broilers and layers at each examined time point was obtained; these data were then used in company with the microarray expression data to identify significant QTE genes of broilers and layers by fitting the following simple linear model to each present gene NIRij and Gij is the NIR and log expression value for gene iat time point j, respectively; significant QTE genes were finally determined as those with significant \u03b2\u03b9 \u2260 0 .where KEGG pathway -44 infor\u03bcl amplification reactions containing 10 \u03bcl of SYBR Green PCR Master Mix, 20 ng cDNA and 0.5 \u03bcM of each primer at the following conditions: 95\u00b0C for 10 minutes for 1 cycle, 40 cycles at 95\u00b0C for 15 seconds and then at 60\u00b0C for 1 minute. Because postnatal 2 weeks to 4 weeks is the period that most divergent growth rate occur between broilers and layers, only RNA samples from postnatal 2 weeks and 4 weeks were used in the qRT-PCR experiment.PCR analyses were performed using the ABI Prism 7500 System. Real-time quantification was employed using SYBR Green PCR Master Mix (ABI). PCR primers were designed using AB PRISM Primer Express 2.0 software. To avoid amplification of contaminated genomic DNA, one of the two primers was placed at or just outside of the exon/exon junction. BLASTN searches were performed to confirm the gene specificity of the primer sequences. All primers for qRT-PCR were listed in Additional file SS): somatostatin; (GHR): growth hormone receptor; (MYH7B): myosin heavy chain 7B; (TNNI1): troponin I type 1; (JNK1): Janus kinase 1; (P38): serine/threonine kinase 38 like; (INCENP): inner centromere protein antigens; (NDE1): nudE nuclear distribution gene E homolog 1; (MUSTN1): musculoskeletal embryonic nuclear protein 1; (CSRP3): cysteine and glycine-rich protein 3; (FHL2): four and a half LIM domains 2; (FGFR2): fibroblast growth factor receptor 2; (FGF16): fibroblast growth factor 16; (HS6ST2): heparin sulphate 6-O-sulfotransferase 2; (HSPGs): heparan sulfate proteoglycans; (MB): myoglobin; (CES1): thiolesterase B; (PDHX): pyruvate dehydrogenase complex, component X; (ALDH1A2): aldehyde dehydrogenase 1 family, member A2; (AER61): glycosyltransferase AER61; (AOX1): aldehyde oxidase 1; (FABP4): fatty acid binding protein 4; (FABP6): intestinal 15 kda protein; (ELOVL 6): homolog, elongation of long chain fatty acids; (PDK4): pyruvate dehydrogenase kinase 4; (OXCT1): 3-oxoacid CoA transferase 1; (COMT): Catechol-O- -methyltransferase; (FBXO22): F-box proteins 22; (FBXO30): F-box proteins 30, (UCH-L1): ubiquitin carboxyl-terminal hydrolase isozyme L1; (HERC4): hect domain and RLD 4; (HERC5): hect domain and RLD 5; (RNF12): ring finger protein 12; (AER61): glycosyltransferase; (FGF1): fibroblast growth factor 1 (acidic); (DKK3): dickkopf homolog 3; (EDNRA): endothelin receptor type A; (EDNRB): endothelin receptor type B; (PTCHD1): patched domain containing 1; (AER61): glycosyltransferase; (LARGE): like-glycosyltransferase; (ACTN3): actinin, alpha3; (MYBPC3): myosin binding protein C, cardiac.(IGF): Insulin-like growth factor; (PI3K): phosphatidylinositol 3-kinase; (mTOR): mammalian target of rapamycin; (AMPK): AMP-activated protein kinase; (ERK1/2): extracellular signal regulated kinase 1/2; (NF-kappa B): nuclear factor-kappa B; (PKC): protein kinase C; and the Natural Growth Rates (unit: \u0394g/g) of broiler and layer chickens are shown as a function of days after hatch (DAH). Red lines, broilers; blue lines, layers; solid lines, absolute growth rate; dashed lines, Natural Growth Rates (NIR).Click here for fileAnnotation and expression values of the 543 differentially expressed genes between broilers and layers. This file provides the probe set annotations and expression values of the 543 differentially expressed genes at all examined time points.Click here for fileList differentially expressed genes between broilers and layers validated by qRT-PCR. This file includes the information of differentially expressed genes confirmed by qRT-PCR. The primer sequences used for qRT-PCR are also included.Click here for fileEnriched GO terms among the 543 differentially expressed probe sets between broilers and layers. This file includes the list of enriched GO terms among the 543 differentially expressed probe sets between broilers and layers, as well as the enrichment degree of each GO term.Click here for fileDifferentially expressed genes in the chicken tyrosine metabolism pathway. This file presents the differentially expressed genes in chicken tyrosine metabolism pathway. The differentially expressed genes within the tyrosine metabolic pathway are highlighted by red ellipses.Click here for fileList of genes with expression patterns correlated with the growth rates of broilers and/or layers (QTE genes). This file includes the list of genes with expression patterns correlated with growth rates of broilers and/or layers. The probe information, annotation and expression values of each gene are included.Click here for fileComparison of enriched GOBP Terms for DE genes vs. common-QTE genes. This file presents the enriched biology process GO terms of the differentially expression genes and the common QTE genes between broilers and layers. Red boxes, enriched GO BP terms for DE genes; Green boxes, enriched GO BP terms for common-QTE genes; Yellow, Shared GO BP terms for both DE-genes and common-QTE genes. The colour saturation degree is positively correlated with the significance of enrichment.Click here for fileGene Ontology analysis of broiler specific QTE genes. Enriched biological process GO terms are highlighted by yellow; the colour saturation degree is positively correlated with the significance of enrichment.Click here for fileGene Ontology analysis of layer specific QTE genes. Enriched biological process GO terms are highlighted by yellow; the colour saturation degree is positively correlated with the significance of enrichment.Click here for fileEnriched KEGG pathways among the 543 differentially expressed genes between broilers and layers. This file lists the enriched KEGG pathways among the 543 differentially expressed genes between broilers and layers. Pathways with FDR less than 0.1 are included.Click here for file"} +{"text": "Recent observations have shown that the pineal hormone melatonin (MLT) may modulate oestrogen receptor (ER) expression and inhibit breast cancer cell growth. On this basis, we have evaluated the biological and clinical effects of a concomitant MLT therapy in women with metastatic breast cancer who had progressed in response to tamoxifen (TMX) alone. The study included 14 patients with metastasis who did not respond (n = 3) to therapy with TMX alone or progressed after initial stable disease (SD) (n = 11). MLT was given orally at 20 mg day-1 in the evening, every day starting 7 days before TMX, which was given orally at 20 mg day-1 at noon. A partial response was achieved in 4/14 (28.5%) patients (median duration 8 months). The treatment was well tolerated in all cases, and no MLT-induced enhancement of TMX toxicity was seen; on the contrary, most patients experienced a relief of anxiety. Mean serum levels of insulin-like growth factor 1 (IGF-1), which is a growth factor for breast cancer, significantly decreased on therapy, and this decline was significantly higher in responders than in patients with SD or progression. This pilot phase II study would suggest that the concomitant administration of the pineal hormone MLT may induce objective tumour regressions in metastatic breast cancer patients refractory to TMX alone."} +{"text": "To the Editor: In 2005 and 2006, the highly pathogenic avian influenza (HPAI) virus subtype H5N1 rapidly spread from Asia through Europe, the Middle East, and Africa. Waterbirds are considered the natural reservoir of low pathogenic avian influenza viruses (Anas querquedula) is the most numerous duck migrating between Eurasia and Africa: \u22482 million gather in the wetlands of Western Africa every northern winter in the period February 7\u201315, 2007. All cloacal and tracheal swabs tested negative for avian influenza virus by real-time reverse transcription\u2013PCR analysis of the matrix gene. One second-year (>9-month-old) female garganey migrated from northern Nigeria to Russia in April\u2013May 2007 , where sThis transcontinental migration path connects several areas of past major HPAI (H5N1) outbreaks . The winDuring spring migration from Nigeria to Russia, the garganey stopped several days in wetlands situated close to areas of past outbreaks in the Danube delta (4 days at a distance of 1\u20134 km from October 2005 outbreaks) and Lake Kus, Turkey (8 days at a distance of 10\u201330 km from October 2005 outbreaks). The occurrence of past outbreaks indicates that the duck used wetlands favorable to HPAI virus (H5N1) transmission as stopover sites. The relatively long stopover periods enabled prolonged contact of migratory ducks with local domestic and wild bird populations or through shared water, thus prolonging the potential for virus transmission. Considering the persistence of infectivity of HPAI virus (H5N1) in aquatic habitats can potentially be transported rapidly by migratory birds across continents. However, the physiologic impact of an HPAI (H5N1) infection on the ability of birds to migrate long distances is still unknown ( A) Migration route of a garganey tracked with satellite telemetry from February through November 2007, and major highly pathogenic avian influenza (H5N1) outbreaks in domestic and wild birds over Europe, the Middle East, and Africa from July 2005 through November 2007. Close-up maps of stopover sites during the spring migration in (B) the Danube River delta in Romania , and (C) Lake Kus in Turkey ."} +{"text": "Of 61 consecutive patients undergoing laparoscopic cholecystectomy, 4 (6.25%) developed abdominal wallhaematomas. This complication of laparoscopic cholecystectomy may occur more commonly than existingliterature suggests, and manifests in the post-operative period (days 2 to 6) by visible bruising, excessive painor an asymptomatic drop in haematocrit. It is readily confirmed by ultrasonography. While no specifictreatment is necessary apart from replacement of significant blood loss, the patient requires reassurance thatthis apparently alarming complication will rapidly resolve."} +{"text": "In contrast, animals vaccinated with only a single CTL epitope and challenged with the same SIVmac251 stock had high levels of viral replication and rapid CTL escape. Unvaccinated na\u00efve animals exhibited a slower emergence of immune escape variants. Thus narrowly directed vaccination against a single epitope resulted in rapid immune escape and viral levels equivalent to that of na\u00efve unvaccinated animals. These results emphasize the importance of inducing broadly directed HIV-specific immunity that effectively quashes early viral replication and limits the generation of immune escape variants. This has important implications for the selection of HIV vaccines for expanded human trials.Successful vaccination against HIV should limit viral replication sufficiently to prevent the emergence of viral immune escape mutations. Broadly directed immunity is likely to be required to limit opportunities for immune escape variants to flourish. We studied the emergence of an SIV Gag cytotoxic T cell immune escape variant in pigtail macaques expressing the Mane-A*10 MHC I allele using a quantitative RT-PCR to measure viral loads of escape and wild type variants. Animals receiving whole Gag expressing vaccines completely controlled an SIV The HIV pandemic continues unchecked and a vaccine is urgently needed. The lack of successful vaccination strategies contrasts starkly with the success of antiretroviral drug therapy. Combinations of drugs that successfully limit HIV replication do not select for drug resistant variants and result in long-term control of infection and near-normal life expectancy. Uniform testing and anti-retroviral treatment is predicted in some models to ultimately control the HIV epidemic Many immune escape mutations (EM), particularly those in conserved proteins such as Gag, are likely to inflict at least some reduction in viral replication capacity \u2013 known as a \u201cfitness cost\u201d. This is demonstrated clearly by reversion of mutations back to the fitter wild type (WT) sequences upon transmission to new hosts not able to mount the same immune response Immune escape can occur at various times after natural infection \u2013 escape variants to strains that induce early CTL responses will tend to appear earlier, sometimes during acute infection The prior generation of effective immune responses through vaccination could effectively quash viral replication and limit opportunities to escape. However, we recently postulated that if replication is sufficiently high, pressure from vaccine-induced response might force immune escape variants to arise early , reducing the flexibility of the overall immune response Macaca nemestrina) involved in previously published SIV vaccine experiments. Macaques were either vaccinated with regimens that either expressed (a) whole SIV Gag protein Mane-A*10 MHC I allele and 4 animals were followed for >20 weeks post infection. Two animals received recombinant viral vectors (influenza A viruses) expressing only the KP9 Gag epitope mac239 Gag-Pol - two animals received a prime/boost regimen with recombinant vaccinia and fowlpox viruses Mane-A*10+ animals from within these vaccine studies were analyzed. The viral loads, CD4 T cell levels and KP9 specific CD8 T cell responses from these studies have previously been reported, and the immune escape profiles from 3 of the na\u00efve animals and the 2 KP9-only vaccinated animals have also been reported. Here we analyze immune escape in all animals and compare the outcomes of infection and timing of escape in these different vaccination scenarios.The 22 pigtail macaques involved in published vaccine studies were analyzed as shown in Mane-A*10 MHC class I allele by pigtail macaques is associated with reduced viral load and reduced disease compared to non-Mane-A*10+ pigtail macaques mac251 viremia was observed in 3 Mane-A*10+ pigtail macaques vaccinated with recombinant viral vectors expressing whole SIV Gag which had the less common escape variant P172S .10 copies/ml) at weeks 7 and 10 after challenge before disappearing.We then employed the qRT-PCR assay on animals with K165R escape to generate serial viral load data for both WT and the K165R KP9 EM variant. In the animals vaccinated whole Gag-expressing vectors, the immune escape variant was not detected at all 2 of 3 animals . In one The failed vaccination approach based on a single epitope (Gag KP9) predictably resulted in the emergence of a CTL escape variant, as previously reported A delayed pattern of CTL escape at the KP9 epitope is observed in the unvaccinated SIV-infected animals. Immune escape was substantially delayed compared to animals vaccinated against KP9 alone, with significant levels of EM virus not appearing until week 5 of infection and not exceeding WT virus until 7\u201310 weeks after infection in the 3 animals with chronic SIV and the K165R mutation .We compared the estimate of the timing of escape starting across the vaccination groups . Immune r\u200a=\u200a\u22120.6390, p<0.0001; for KP9-vaccinated animals r\u200a=\u200a\u22120.749, p\u200a=\u200a0.0149; and for na\u00efve animals r\u200a=\u200a\u22120.523, p\u200a=\u200a0.0018. Spearman correlation).We calculated the rates of immune escape across all animals and timepoints as previously described mac251The control of viremia and prevention of immune escape at the KP9 CD8 T cell epitope by whole Gag-expressing vaccines suggested that immunity to other regions within Gag may have been generated. Indeed Gag-specific T cell immunity is linked to control of viremia both in pigtail macaques infected with SIVHIV-infected subjects who spontaneously control viremia are likely to mount effective combinations of immune responses. Understanding and inducing such effective immunity by vaccination is a priority. We found that inducing CTL responses to a single epitope resulted in the early selection of immune escape species and no control of viremia, consistent with previous macaque vaccine studies The use of mono-epitope vaccine strategies is likely to be particularly damaging: the recall response following viral challenge will be primarily directed towards the escaping epitope. Subdominant responses, which could otherwise help facilitate viral control Mane-A*10 MHC I allele which restricts the dominant KP9 Gag response, alternative MHC alleles restricting other useful T cell responses in this outbred group could confound our data. More extensive MHC typing found none of the vaccinated macaques expressed the Mane-B*10 or A*17 alleles previously shown to present the dominant KW9 or AF9 Gag epitopes Several future studies should assist in refining this model of CTL-based control of primate lentiviruses. Although we studied 22 macaques in total, only smaller subsets of these were vaccinated and larger studies will help validate this work. Further, although all macaques expressed the An additional area requiring further work is that different vaccine vectors will likely play an important role in the immune responses generated. Although we observed a similar magnitude of the KP9 responses after challenge despite different vaccine vectors studied, we did not examine the many other aspects of the quality of the CD8 T cell responses generated by vaccination as shown in Mane-A*10 by reference strand-mediated conformational analysis and sequence-specific primer PCR as described mac251 IV 40 TCID50. Four animals did not receive any SIV-expressing vaccines. Two animals received recombinant influenza A viruses expressing only the KP9 Gag epitope mac239 Gag-Pol mac239 Gag-Pol Mane-A*10+ animals from within these vaccine studies were analyzed.We studied 22 KP9-specific immune responses were studied on serial blood samples using a Mane-A*10/KP9 tetramer and flow cytometry as previously described 4 cycler. Analysis was performed using Eppendorf Realplex software. Baselines were set 2 cycles earlier than real reported fluorescence and threshold value was determined by setting threshold bar within the linear data phase. Samples amplifying after 40 cycles were regarded as negative, and corresponded to <1.5-Log10 SIV RNA copies/ml of plasma. Analyses of the rate of immune escape were performed as previously described by calculating the change in the viral species over time To quantify virus levels of WT or EM quasispecies at the KP9 epitope we employed a discriminatory real-time PCR assay as described To analyze the impact of breadth of T cell immunity to Gag (i.e. other than KP9-specific responses), we performed intracellular cytokine staining assays for T cell activation using 5 pools of 25 overlapping 15mers peptides spanning the entire SIV Gag protein as previously described"} +{"text": "The leucocyte migration and guinea-pig macrophage migration procedures were used to assess cell mediated, tumour directed immune reactions in patients with mammary carcinoma undergoing simple mastectomy with or without post-operative irradiation. Forty-seven per cent of patients reacted to autologous tumour antigens and 40% to allogeneic antigens when tested 7 days after operation; 23% reacted to autologous antigens at 2 months, 19% at 6 months and 34% at 1 year after surgery. Reactions to benign tissue fractions were rare. Better discrimination between test and control subjects was obtained when 3000 g sediments rather than nuclei-depleted homogenates (extracts) were used. Irradiation 3-7 weeks post-operatively did not depress the in vitro response at 2 months and yielded a higher rate of positive reactions at 6 months. Correlations of serial LMT responses with certain clinical findings are discussed."} +{"text": "Reaction of aromatic/heterocyclic sulfonamides containing a free amino group with triflicanhydride afforded compounds possessing trifluoromethanesulfonamido moieties in their molecule. The Zn(II) and Cu(II) complexes of these new sulfonamides were prepared and characterized by standard procedures . The new derivatives showed good inhibitory activity against three isozymes of carbonic anhydrase (CA), i.e., CA I, II and IV."} +{"text": "Non-synonymous single nucleotide polymorphisms (SNPs) within vital DNA repair genes may cause reduction of activity leaving the genome unrepaired resulting in genomic instability and cancer.The present endeavour involved study on the association of the SNP rs13181 (Lys751Gln/A18911C) in the Nucleotide Excision Repair (NER) pathway gene ERCC2 with the risks of Squamous Cell Carcinomas of the Head and Neck (SCCHN) and Breast cancer using a case-control based association study among 685 (400 controls and 285 SCCHN-affected cases) and 395 ethnically-matched samples, respectively from north India using Polymerase Chain Reaction followed by Restriction Fragment Length Polymorphism (PCR-RFLP) analysis.Results showed significant association of rs13181 homozygous mutant (CC) , heterozygous (AC) and combined mutant (AC + CC) genotypes with predisposition to Breast cancer. Statistically significant increase in SCCHN risk was also associated with the mutant genotypes of rs13181 (ERCC2), viz. homozygous mutant (CC) , heterozygous (AC) and combined mutant (AC + CC) genotypes.The results of this case-control study indicate that the polymorphism rs13181 might be a risk factor for predisposition towards SCCHN and breast cancer among north Indian subpopulations. Breast cancer is the primary subtype of cancer leading to death among women in developing countries. 13% out of the 58 million deaths worldwide in the year 2005 were caused due to cancer which included 502,000 deaths per year due to breast cancer. Well-established risk factors ascribed to breast cancer include early menarche, late menopause, age of first child's birth, nulliparity and family history (FH) [Cancer is a genetic disease resulting from gradual accumulation of changes in the DNA that activate proto-oncogenes and inactivate tumour-suppressor genes leading to genetic instability which is further aggravated by DNA damage and errors made by the DNA maintenance and repair machinery . Many caorldwide with higorldwide . SCCHN aory (FH) .DNA repair is considered to play a key role in cancer susceptibility whereby some individuals are at very high risk of cancer due to SNPs in crucial DNA repair genes -15. InacThe gene ERCC2 encodes the ERCC2/Xeroderma pigmentosum Type D (XPD) protein, which is one of the seven genetic complementation groups that forms an essential component of the Nucleotide excision repair (NER) pathway, a major DNA repair pathway that removes photoproducts from UV radiation and bulky adducts from a huge number of chemicals, cross-links and oxidative damage through the action of 20 proteins and several multiprotein complexes . XPD is In the present study, genetic association of the nonsynonymous SNP rs13181 with the risks of Breast cancer and Squamous Cell Carcinomas of the Head and Neck (SCCHN) was analysed using Polymerase Chain Reaction followed by Restriction Fragment Length Polymorphism (PCR-RFLP) in a subpopulation cluster-matched case-control based study among north Indians.Blood samples (2 ml each) were collected following written informed consent from 168 Breast cancer patients and 285 SCCHN patients, following histopathological and cytological confirmation, from different parts of north India undergoing treatment at Lucknow Cancer Institute (LCI) and Sanjay Gandhi Post Graduate Institute of Medical Sciences (SGPGIMS) between September, 2005 and June, 2008. 400 unrelated ethnically-matched cancer-free blood donors from the north Indian states of Uttar Pradesh and Uttarakhand were included in the current study as healthy normal controls for cancer association studies. All the subjects inducted in this study belonged specifically to the Caucasoid morphological subtype and IndoDNA was isolated from blood samples of SCCHN and Breast cancer cases and controls using QIAamp DNA Blood Midikit (Qiagen Inc.) following manufacturer's protocol, quantitated using spectrophotometer and stored at -20\u00b0C. and UCSC In-silico PCR to eradicate the possibility of amplification of any non-specific DNA sequences and synthesized commercially.Reference sequence of the gene ERCC2 and information on coding regions (CDS) were retrieved from NCBI's sequence databases. The primers 5' CCCCCTCTCCCTTTCCTCTGTTC 3' (Forward Primer) and 5' GGACCTGAGCCCCCACTAACG 3' (Reverse Primer) were designed for the present study on the SNP rs13181 (ERCC2) using PrimerSelect module of Lasergene v6.0 software (DNAStar). The primer sequences were verified using NCBI BLAST Gradient PCR reactions were performed for standardization of DNA amplification conditions and optimization of annealing temperature for the set (forward + reverse) of primers. Briefly, the primer set was used to amplify a standard DNA template at different annealing temperatures (with increment of approximately 2\u00b0C) and the temperature at which highest amount of PCR product was formed was considered the optimum annealing temperature for further PCR reactions.2, 0.2 mM dNTPs, and 15 pmol of each primer. Thermal cycling conditions were as follows: initial denaturation step at 95\u00b0C for 10 min, 31 cycles of PCR consisting of denaturation at 94\u00b0C for 1 min, annealing at 63.0\u00b0C for 1 min and extension at 72\u00b0C for 1 min, followed by a final extension step at 72\u00b0C for 5 min. The reaction was held at 4\u00b0C. The PCR products were visualized by electrophoresis on 1.2% agarose gel and stored at 4\u00b0C. For gel electrophoresis, 5 \u03bcl of the amplified product was mixed with 1 \u03bcl of 6\u00d7 gel loading buffer and resolved on 1.2% agarose gel in TAE buffer at 85 volts for 1 1/2 hrs. 100 bp DNA markers (New England Biolabs) were run with the amplified products as reference.All PCR reactions were performed in 200 \u03bcl transparent PCR tubes (Axygen Scientific Pvt. Ltd.) on a peltier-based thermal cycler using reagents from Fermentas Life Sciences in a total reaction volume of 50 \u03bcl containing nearly 100 ng genomic DNA, 1.5 U Taq polymerase in 1\u00d7 PCR buffer, 1.5 mM MgCl, an on-line tool for SNP-RFLP analysis. The 413 bp PCR product was subjected to restriction digestion using PstI following optimum reaction conditions as per manufacturer's protocols. The digestion products were visualized by electrophoresis on 3% agarose gel for RFLP analysis and the genotypes were inferred from the number of bands observed in the gel. The homozygous wild type (AA) genotype generated a single band of 413 bp upon restriction digestion, the homozygous mutant genotype (CC) produced two bands of 322 bp and 91 bp, while the heterozygous genotype (AC) was inferred by the presence of all the three bands upon visualisation on agarose gel following restriction digestion using the enzyme PstI.The restriction enzyme PstI (Fermentas Life Sciences) was selected for PCR-RFLP studies using SeqBuilder module of Lasergene 6.0 (DNAStar) and WATCUT among cases and controls separately, comparing the observed allele counts with that of the expected, by means of Goodness-of-fit Chi square test at df (degrees of freedom) = 1. 3 \u00d7 2 Contingency Chi-square test was performed to verify overall association of the genotypes between cases and controls. Odds ratios (OR), relative risk (RR) and corresponding 95% confidence intervals (CI) were estimated to ascertain association of individual genotypes with SCCHN and Breast cancer risks. Logistic regression was performed to calculate adjusted ORs for subsequent analysis of potential risk factors like gender, smoking, Tobacco chewing and pan masala. All statistical tests were two-sided.Maximum likelihood estimates of descriptive statistics like allele and genotype frequencies were calculated using Microsoft Excel. Further statistical analysis of data was performed using the computer softwares Statistical Package for the Social Sciences (SPSS) version 16.0 and GraphPad Prism version 5.0. Hardy-Weinberg Equilibrium (HWE) was tested online using Hardy-Weinberg Equilibrium Calculator HWE for genotype distributions were 0.2488 among controls. Genotype and allele frequencies for the loci rs13181 (ERCC2) among Breast cancer cases and normal healthy female controls have been provided in Tables Genotype results were successfully obtained among 215 female controls and 155 breast cancer cases. ChisquareStatistically significant association with breast cancer susceptibility was observed for the mutant genotypes of the polymorphism rs13181 in the gene ERCC2 viz. homozygous mutant (CC) , heterozygous (AC) and combined mutant (AC + CC) . Results of association studies between the mutant genotypes and breast cancer risk are represented in terms of corresponding Odds ratios in Table HWE for genotype distributions was 0.345 among controls. The genotype and allele frequencies of the SNP rs13181 (ERCC2) among SCCHN cases and healthy control subjects are provided in Tables Genotype results were successfully obtained among 385 healthy unaffected control subjects and 275 SCCHN-affected cases for rs13181 (ERCC2). ChisquareThe ERCC2/XPD protein functions as an ATP-dependent 5'-3' helicase joint to the basal TFIIH complex and participates in the local unwinding of DNA helix to allow RNA transcription machinery to access the promoter and to permit the NER machinery to access the lesion ,52. SeveLiteratures evaluating the risk of rs13181 (ERCC2/XPD) polymorphism with the risk of Breast cancer have been controversial. Although some studies found no correlation between this polymorphism and breast cancer risk ,56-59, sOn the other hand, only two studies have been found to have been conducted so far on the assessment of the risk of SCCHN associated with rs13181 (ERCC2) mutant genotypes. Both of these studies exhibited significant positive association with SCCHN risk among non-Hispanic white subjects and the south Indian population, respectively ,62. CorrThe results of the present investigation indicate that the polymorphism rs13181 might be a risk factor for predisposition towards SCCHN and Breast cancer among north Indian subpopulations. The data generated from this study may have wide-ranging applications for further epidemiological and public health related research on the Indian population.The degree of susceptibility to cancers is hypothesised to be the final product of a mishmash of high-risk genetic polymorphic variants or SNPs in a subset of medium and low penetrance genes like DNA repair genes which, even in the absence of the highly penetrant variant cancer-associated alleles, may increase the degree of susceptibility towards cancers a few fold thus having a major impact on the population incidence of cancer . TherefoThe authors declare that they have no competing interests.AKM conceived of the study, participated in sample collection, carried out the molecular genetic studies, performed PCR-RFLP analysis, conducted the statistical analysis and drafted the manuscript. NS participated in sample collection. VKG offered clinical support and provided cancer samples. RC and MS carried out histopathology on the cancer samples. SKR supervised the study, participated in its conception, design and coordination and reviewed the manuscript. All authors read and approved the final manuscript."} +{"text": "In this study, we analysed immunocytochemically p53 expression in first primary and second primary cancers from 25 head and neck cancer patients (HNCPs) with multiple malignancies in comparison with oncoprotein expression in tumour tissues from 25 historical HNCP controls with single cancer in a match-paired analysis. Moreover, we investigated bleomycin-induced chromosome fragility in both groups of HNCPs and in 21 additional healthy controls. Thirty-nine out of 75 tumour specimens analysed (52%) showed positive p53 immunostaining. Eleven out of 25 (44%) from single cancer patients and 28 out of 50 (56%) tumours from HNCPs with multiple malignancies were p53 positive. In the group of multiple primary cancers, nine patients (36%) showed positive staining of both first and second primaries, whereas six (24%) had positive labelling of first primary cancer but not of the subsequent second primary, four (16%) patient showed p53 expression only in the second primary cancer and six (24%) patients showed no p53 immunoreactivity in both tumours. Chromosomal analysis demonstrated a higher sensitivity to clastogens of HNCPs with multiple tumours than of HNCPs with a single cancer (P < 0.01), and a significant correlation between chromosome fragility and p53 overexpression (P < 0.01) only in HNCPs with multiple malignancies more than in those with single head and neck cancer (P = 0.11). Moreover, we found that patients with p53-positive staining of both first and second primaries showed a statistically significant higher mutagen sensitivity than those with a single p53 immunoreactive tumour or those in whom both cancers were p53 negative (P < 0.01). Our data suggest that subjects with increased susceptibility to carcingogens after exposure to tobacco or alcohol are at higher risk for multiple cancers in which one of the most common genetic events is aberrant p53 expression."} +{"text": "P = 0.09). Relapsed patients had a longer TTD than patients with progressive disease (P = 0.002). Early relapse led to a shorter TTD than late relapse (P = 0.005). Median TTD was 14 months for patients who underwent salvage therapy and 2 months for untreated cases (P < 0.00001). A multivariate analysis showed an independent prognostic role for salvage therapy and time to relapse. Age and type of failure had no predictive value. Salvage therapy significantly improves outcome and, possibly, quality of life. As many different treatments were used conclusions cannot be made regarding an optimal treatment schedule. \u00a9 1999 Cancer Research CampaignFailure after first-line treatment was reported in 35\u201360% of immunocompetent patients with primary central nervous system lymphoma (PCNSL). There are currently no reports focusing on salvage therapy. This review analyses prognostic factors and the efficacy of salvage therapy by focusing on data from papers reporting results of first-line treatment in 355 cases. The study group consisted of 173 patients presenting treatment failure. The interval between failure and death (TTD) was compared for age at relapse (\u226460 vs >60 years), type of failure (relapse vs progression), time to relapse (\u226412 vs >12 months) and salvage treatment (yes vs no). Median TTD was similar in younger and older patients ("} +{"text": "Erythropoietin (EPO), a hematopoietic cytokine, enhances neurogenesis and angiogenesis during stroke recovery. In the present study, we examined the effect of EPO on oligodendrogenesis in a rat model of embolic focal cerebral ischemia.Recombinant human EPO (rhEPO) at a dose of 5,000 U/kg (n\u200a=\u200a18) or saline (n\u200a=\u200a18) was intraperitoneally administered daily for 7 days starting 24 h after stroke onset. Treatment with rhEPO augmented actively proliferating oligodendrocyte progenitor cells (OPCs) measured by NG2 immunoreactive cells within the peri-infarct white matter and the subventricular zone (SVZ), but did not protect against loss of myelinating oligodendrocytes measured by cyclic nucleotide phosphodiesterase (CNPase) positive cells 7 days after stroke. However, 28 and 42 days after stroke, treatment with rhEPO significantly increased myelinating oligodendrocytes and myelinated axons within the peri-infarct white matter. Using lentivirus to label subventricular zone (SVZ) neural progenitor cells, we found that in addition to the OPCs generated in the peri-infarct white matter, SVZ neural progenitor cells contributed to rhEPO-increased OPCs in the peri-infarct area. Using bromodeoxyuridine (BrdU) for birth-dating cells, we demonstrated that myelinating oligodendrocytes observed 28 days after stroke were derived from OPCs. Furthermore, rhEPO significantly improved neurological outcome 6 weeks after stroke. In vitro, rhEPO increased differentiation of adult SVZ neural progenitor cells into oligodendrocytes and enhanced immature oligodendrocyte cell proliferation.Our in vivo and in vitro data indicate that EPO amplifies stroke-induced oligodendrogenesis that could facilitate axonal re-myelination and lead to functional recovery after stroke. Oligodendrocytes are the myelin-forming glial cells in the adult brain and are highly vulnerable to ischemic insult Erythropoietin (EPO), a hematopoietic cytokine, facilitates oligodendrocyte maturation in vitro P<0.05) improved neurological outcome compared with saline treated rats 6 weeks after stroke (P>0.05) different between rats treated with saline and rhEPO (34.4\u00b15.6%).To test the restorative effect of recombinant human EPO (rhEPO) on stroke, rhEPO was intraperitoneally administered daily for 7 days starting 24 h after stroke onset. Neurological deficits were examined before the treatment and 2 and 6 weeks after stroke by means of modified neurological severity score (mNSS) and foot-fault test. All rats exhibited severe deficits measured by mNSS and foot-fault test 24h after middle cerebral artery (MCA) occlusion and there were no significant differences among the groups . Althougr stroke , which iAforementioned data suggest that the effect of EPO on improvement of functional outcome is not primarily resulted from the neuroprotective effect. To examine the effect of EPO on oligodendrocytes in the ischemic brain, we measured OPCs and myelinating oligodendrocytes 7, 28 and 42 days after embolic ischemia. OPCs were identified by NG2 positive cells, while myelinating oligodendrocytes were detected by cyclic nucleotide phosphodiesterase (CNPase) or myelin basic protein (MBP) immunoreactive cells The adult SVZ generates OPCs that disperse throughout the corpus callosum and striatum Twenty-eight and 42 days after MCAo, ischemic rats treated with rhEPO exhibited a significant increase in CNPase and MBP immunoreactivity along the peri-infarct corpus callosum and striatum compared to the ischemic rats treated with saline and 4. TFor both rhEPO treated and saline control rats, the number of NG2 positive cells in the peri-infarct region and the SVZ decreased 28 and 42 days after MCAo compared with the number at 7 days after stroke . HoweverOur observation that rhEPO increased mature oligodendrocytes prompted us to examine the effect of EPO on myelinated axons. We performed Bielschowsky and Luxol fast blue staining which detects myelinated axons and neurofilament-H (NF-H) immunostaining which labels axons To verify our in vivo findings, we performed in vitro experiments using primary SVZ cells harvested from the adult rat and an oligodendrocyte cell line (N20.1). Adult rodent SVZ cells contain oligodendrocyte progenitor cells that migrate into the white matter In addition to the SVZ, many OPCs are present in white matter The present study demonstrates that administration of rhEPO 24 h after embolic MCAo induced sustained OPC proliferation in the peri-infarct white matter and the SVZ. In addition, rhEPO treatment substantially amplified myelinating oligodendrocytes and increased myelinated axons in peri-infarct white matter, which was associated with substantial improvement of functional outcome 6 weeks after stroke. These data suggest that EPO amplifies stroke-induced oligodendrogenesis and axonal remodeling that may contribute to functional recovery after stroke.Oligodendrocytes are the myelin-forming glial cells in the adult brain Studies in CNPase knockout mice indicate that CNPase expressing cells are required to support axonal integrity NG2 positive cells are generally considered as OPCs, although NG2 can be induced in activated microglia All experimental procedures were carried out in accordance with the NIH Guide for the Care and Use of Laboratory Animals and approved by the Institutional Animal Care and Use Committee of Henry Ford Hospital .Male Wistar rats weighing 350\u2013400 g were subjected to embolic MCA occlusion, as previously described To test the effect of rhEPO on stroke, rhEPO at a dose of 5,000 units/kg was intraperitoneally administered daily for 7 days starting 24 h after stroke onset (n\u200a=\u200a18). As a control group, the same volume of saline was administered to ischemic rats (n\u200a=\u200a18) with the identical protocol described above. For mitotic labeling, BrdU (100 mg/kg) was administered daily for 7 days starting 24 h after stroke onset.Lentiviral labeling: To examine whether SVZ neural progenitor cells migrate to the ischemic boundary region, SVZ cells were labeled with a lentivirus-GFP vector that was constructed, as previously described All functional outcome tests were performed by observers blinded to the treatments 1, 14, and 42 days after onset of MCA occlusion.Rats were tested for motor, sensory, reflex, and balance dysfunctions with the mNSS A modified foot-fault test was employed to measure forelimb placement dysfunction Rats were sacrificed 7, 28 or 42 days after stroke. The brains were removed and consecutive coronal sections at bregma -0.4 to -1.4 mm were prepared. Infarct volume was measured on 7 hematoxylin and eosin (H&E) stained coronal sections using the microcomputer imaging device (MCID) system , as previously described OPCs were identified by staining with antibodies against NG2 and O4 . CNPase is a prenylated myelin protein and MBP is an abundant protein component of the myelin sheath, which are highly expressed in mature oligodendrocytes Five coronal sections (8 \u00b5m/section) spaced as 100 \u00b5m intervals per staining were used from each rat and these coronal sections were within the territory supplied by the MCA at bregma \u22120.4 to \u22121.4 mm Coronal sections double stained with antibodies against NF-H and CNPase or MBP were imaged using Zeiss confocal microscopy (Zeiss LSM 510 NLO).4 cells/ml in the presence of growth medium To directly test whether EPO regulates oligodendrogenesis, neural progenitor cells were isolated from the SVZ of adult rats (n\u200a=\u200a3), as previously described To further investigate the effect of EPO on oligodendrocyte cell proliferation, we employed a mouse premature oligodendrocyte cell line , which was obtained from mouse primary cultures of oligodendrocytes conditionally immortalized by transformation with a temperature-sensitive large T-antigen P<0.05. All values are presented as mean \u00b1 SE.Data were evaluated for normality. Behavioral data were evaluated with one-way analysis of variance (ANOVA) followed by Student-Newman\u2013Keuls test. Two sample t-tests were used to compare the group difference on histological outcome if data were normal, otherwise nonparametric (Wilcoxon) test was considered. Statistical significance was set at"} +{"text": "P< 0.05) and maintained appetite (P< 0.05). Serum and urine concentrations of basic fibroblast growth factor (bFGF), TNF-\u03b1 and VEGF were measured during treatment and higher levels were associated with progressive disease. Thalidomide was well tolerated: Two patients developed WHO Grade 2 peripheral neuropathy and eight patients developed WHO grade 2 lethargy. No patients developed WHO grade 3 or 4 toxicity. Further studies evaluating the use of Thalidomide at higher doses as a single agent for advanced renal cancer and in combination with biochemotherapy regimens are warranted. \u00a9 2000 Cancer Research CampaignTo grow and metastasize, solid tumours must develop their own blood supply by neo-angiogenesis. Thalidomide inhibits the processing of mRNA encoding peptide molecules including tumour necrosis factor-alpha (TNF-\u03b1) and the angiogenic factor vascular endothelial growth factor (VEGF). This study investigated the use of continuous low dose Thalidomide in patients with a variety of advanced malignancies. Sixty-six patients with advanced measurable cancer received Thalidomide 100 mg orally every night until disease progression or unacceptable toxicity was encountered. Three of 18 patients with renal cancer showed partial responses and a further three patients experienced stabilization of their disease for up to 6 months. Although no objective responses were seen in the other tumour types, there were significant improvements in patients' sleeping ("} +{"text": "We have used primary endocrine therapy for 61 elderly women with operable breast cancer (median age 77 years). Eleven patients (18%) had complete and 24 (39%) partial tumour regression, 12 (20%) had stable disease for a minimum of six months and 14 (23%) no response. Salvage surgery was undertaken in the 14 with no response and 8/9 with progressive disease following initial response, thus samples were available from relapse patients only. Assays for EGFr (two point radioreceptor assay) and oestrogen receptors (ER) were performed on 20/22 patients. Ten of these 20 tumours were EGFr+ (greater than 10 fmol mg-1 binding) and 9/13 patients progressing within six months had EGFr+ tumours. 15/22 were available for ER evaluation and there was no such association with ER status. EGFr status was also associated with early recurrence after surgery and death in the endocrine failure group (P less than 0.005 and P less than 0.05 respectively). Of a control population of 33 patients (median age 72 years) treated by primary surgery, only 6 were EGFr+. In this group early relapse was predicted by EGFr status, but not by ER status . There was a significantly higher proportion of EGFr+ tumours in the endocrine failure group compared with the control population (P less than 0.001). EGFr status is a marker for rapid early progression on primary endocrine therapy and the development of non-excisional methods of EGFr analysis would allow better directed therapeutic decisions."} +{"text": "TP53 mutation is highly predictive of complete response to high-dose epirubicin/cyclophosphamide chemotherapy. In these tumours with an altered control of genomic stability, accumulation of chemotherapy-induced genetic alterations may contribute to cell death and account for complete response. To explore the effects of chemotherapy on stability of the tumour genome, allelic profiles were obtained from microdissected tumour samples before and after chemotherapy in 29 unresponsive breast cancers (9 with TP53 mutation). Ninety-four per cent allelic profiles remained unchanged after treatment. Interestingly, 11 profiles (6%) showed important changes after treatment; allelic imbalances significantly increased (four cases) or decreased (seven cases) after chemotherapy in three distinct experiments, two of which using laser microdissected tumour cells. These genetic changes were not linked to the TP53 status, but one tumour showed complete disappearance of TP53-mutated cells in the residual tumour after treatment. Altogether, these observations carry important implications for the clonal evolution of breast cancers treated with DNA-damaging agents, as they point both to the importance of tumour heterogeneity and chemotherapy-driven selection of subclones.In advanced breast cancers, Breast cancers are a heterogeneous group of tumours. Whereas most advanced breast cancer patients receive neoadjuvant chemotherapy, less than 20% of them will fully benefit from this type of treatment and reach complete pathological response, which is closely linked with longer survival more than 15% of the yeast colonies were red and (ii) analysis using the split versions of the test could identify the defect in the 5\u2032 or 3\u2032 part of the gene, confirming the initial determination and (iii) sequence analysis from mutant yeast colonies could identify an unambiguous genetic defect .After tumour RNA extraction , TP53 stTP53 status with the yeast functional assay in all 29 tumours before and after chemotherapy, whenever sufficient frozen material was available for RNA extraction (18 tumours).We determined the TP53 mutation before treatment, frozen tumour tissue was available after treatment in six cases and used to perform the yeast functional test as well as TP53 gene sequencing. For the other three mutated cases, we tried to use formalin-fixed paraffin-embedded tissue, but failed to obtain good enough RNA to perform the yeast functional assay or long enough DNA fragments (>300\u2009bp) to perform gene sequencing.For the nine cases with For all nine mutated cases, TP53 immunostainings on paraffin-embedded samples were obtained before and after treatment using DO-7 monoclonal mouse anti-human antibody , diluted 1/50 in an automated immunostainer . The percentage of stained cells (nuclear staining) was noted.\u03bcm thick paraffin sections were spread on membrane-coated slides and stained with H&E (\u03bcl of buffer (50\u2009mM Tris-HCl (pH 7.5), 1\u2009mM EDTA, 0.5% Tween 20, 0.2\u2009mg\u2009ml\u22121 proteinase K). After an incubation of 24\u201348\u2009h at 56\u00b0C, proteinase K was inactivated at 95\u00b0C for 10\u2009min. No further DNA extraction was performed before PCR analysis for these microdissected samples , EDTA 1\u2009mM) were also analysed with DNA extracted from frozen whole tumour tissue. Briefly, frozen tumour sections were immersed in a buffer containing 8\u2009M MgCl2, 0.2\u2009mM dNTP, 0.2\u2009\u03bcM labelled forward primers or VIC\u2122 for tumour) and 0.2\u2009\u03bcM non-labelled reverse primers. The PCR final volume was 20\u2009\u03bcl. Thirty-five cycles of PCR were performed.For microdissected cells, a volume of lysis buffer accounting for 500 cells was added in each PCR vial. For DNA extracted from blood cells, 10\u2009ng DNA was used for each PCR. The following 10 microsatellite dinucleotide repeats were used : D3S1573After denaturation, the PCR products were run on ABI PRISM 310 Genetic Analyzer. The analysis of the migration data were performed with Genescan 3.1 software (Applied Biosystems).Fluorescent allelic ratios obtained from microdissected tumour tissue were compared with fluorescent allelic ratios obtained from control DNA, allowing the measurement of allelic imbalances (AI). Loss of heterozygosity (LOH) was defined as AI greater than 50% for one tumour allele.AI observed after chemotherapy were then compared to AI observed before chemotherapy, showing locus with unchanged AI, decreased AI or increased AI. Decreased or increased AI were considered significant only when allelic peak heights differed by at least 50%. This threshold was chosen to minimize the risk of false-positive results induced by PCR variations. All these significant pre-post-treatment differences were verified with a second round of PCR performed with microdissected tissue and a third round of PCR performed with frozen whole tumour DNA.Among a total of 290 \u2018before chemotherapy\u2019 PCR (10 markers performed in 29 tumours), 63 allelic profiles were homozygote (non-informative) and 15 PCR could not be analysed due to technical issues. The rates of informativity for these markers were similar to those expected from databases and 3. TAmong a total of 290 \u2018after chemotherapy\u2019 PCR , 63 allelic profiles were homozygote (non-informative) and 24 PCR could not be analysed due to technical issues. The 203 informative profiles revealed 78 (38%) LOH and 125 (62%) retentions of heterozygosity. and 3* cells).Altogether, 191 pairs of PCR were informative and analysable before and after chemotherapy. In 180 pairs (94%), allelic profiles before and after chemotherapy were not different or after treatment .The overall rates of LOH at all loci were similar in tumours with or without TP53 intragenic IGP53 locus before treatment was observed in 4 out of these 9 tumours with TP53 mutation (44%) and in 5 out of 20 tumours without TP53 mutation (20%). Eighteen tumours had sufficient post-treatment frozen material to allow analysis with the yeast functional assay after chemotherapy. Among them, all 12 tumours that did not bear TP53 mutation before treatment did not show post-treatment TP53 mutation.LOH at the TP53 mutation before treatment had good quality post-treatment tissue available and have been further analysed for post-treatment TP53 status or a single small (2\u2009mm) residual focus of tumour cells (patient 8).Six out of nine tumours with 3 status . Among tTP53 status of the tumours.The distribution of the 11 post-chemotherapy profile changes was not linked to any other clinicopathological parameter and was not linked to the pre-treatment We found that genetic changes are detectable early after chemotherapy in breast cancers.All 29 patients had poor prognosis breast tumours treated with the same dose\u2013dense epirubicin/cyclophosphamide regimen. The large amount of tumour tissue obtained before and after (mastectomy) treatment allowed reliable assessment of response to treatment. Obtaining such homogeneous pre- and post-chemotherapy tissue samples is difficult and explains why we could study only 29 patients.Advanced breast cancers are often high grade with necrotic areas, and become more heterogeneous after treatment, with inflammatory infiltrate, oedema, fibrosis and necrosis. To overcome tissue heterogeneity, we used laser tissue microdissection, which allows a targeted sampling of tumour cells with very little contamination. In this regard, 20/29 tumours in our study had at least one allelic loss greater than 80%, indicating that tumour tissue samples were made of almost pure tumour cells. Although laser-microdissection is a highly efficient sampling method, it yields limited amounts of tissue material, and we took special care of sampling at least 5000 cells for each tumour. Indeed, allelic profiling must take into account allelic dropout artefacts that are artificial allelic imbalances occurring when DNA concentration is too low , the TP53 mutation found before chemotherapy was not found after that showed a low post-treatment stromal cell density as well as an important decrease of the proportion of cells with TP53 LOH is clearly in favour of clonal selection. Yet, whatever the mechanism, these treatment-induced events should favour the emergence of therapy-resistant tumour subclones and subsequent tumour recurrences.In this study, we demonstrate that genetic changes do occur early after chemotherapy in breast carcinomas. The most likely explanation for our observations is chemotherapy-driven selection of tumour subclones: breast cancer tumour cells are well known to be highly heterogeneous ("} +{"text": "Hyperbaric oxygen (HBO) has been proposed to reduce tumour hypoxia by increasing the dissolved molecular oxygen in tissue. Using a non-invasive magnetic resonance imaging (MRI) technique, we monitored the changes in MRI signal intensity after HBO exposure because dissolved paramagnetic molecular oxygen itself shortens the T1 relation time. SCCVII tumour cells transplanted in mice were used. The molecular oxygen-enhanced MR images were acquired using an inversion recovery-preparation fast low angle shot (IR-FLASH) sequence sensitizing the paramagnetic effects of molecular oxygen using a 4.7 tesla MR system. MR signal of muscles decreased rapidly and returned to the control level within 40 min after decompression, whereas that of tumours decreased gradually and remained at a high level 60 min after HBO exposure. In contrast, the signal from the tumours in the normobaric oxygen group showed no significant change. Our data suggested that MR signal changes of tumours and muscles represent an alternation of extravascular oxygenation. The preserving tumour oxygen concentration after HBO exposure may be important regarding adjuvant therapy for cancer patients. \u00a9 2000 Cancer Research Campaign"} +{"text": "Sir,WHO has classified adolescent age group as 10-19 years.(th percentile for age on at least three occasions (WHO)..3 Nationad gender. Adult hyMoreover in updated Task Force Report on high blood pressure in children and adolescents, National High Blood Pressure Education Programme (NHBPEP) Coordinating Committee has given hypertension criteria for 1-17 years of age.Hence if one wishes to conduct a survey strictly on \u2018adolescent hypertension\u2019 what age group the researcher should select?"} +{"text": "Patients with an elevated level of urokinase plasminogen activator (uPA) in breast cancer tissue have an adverse prognosis. This study evaluated the prognostic relevance of uPA detection in disseminated tumour cells in bone marrow. Bone marrow was sampled intraoperatively from both iliac crests in 280 patients with primary breast cancer. Interphase cells were enhanced and stained immunocytologically with two antibodies: 2E11, which detects TAG 12--a tumour-associated glycoprotein typically expressed by almost all breast cancer cells--and the anti-uPA antibody HD-UK9. Thirty-five of the 2E11-positive women developed metastatic disease (median follow-up time 44 months). Of these, most were uPA positive and only 12 were uPA negative. Patients with uPA-positive cells in bone marrow had a significantly shorter metastasis-free interval (36 months) than women who were uPA negative (44.5 months). The worst prognosis was seen in patients positive for both markers (29.5 months), followed by those who were uPA negative and 2E11 positive (37 months). The detection of uPA on disseminated tumour cells characterizes a subgroup of patients with an even worse prognosis, who should undergo more aggressive adjuvant systemic therapy. For the first time, it was possible to evaluate an important qualitative parameter involved in the process of breast cancer metastases."} +{"text": "Thus, the coordination environment of the CaII ions is composed of six O atoms belonging to the phosphoryl and sulfonyl groups of two chelate rings and two additional O atoms of two bridging sulfonyl groups. The coordination polyhedron of the central atom (2 symmetry) has a distorted octa\u00adhedral geometry.The crystal structure of the title calcium complex, [Ca(C DOI: 10.1107/S1600536809032875/bq2148Isup2.hkl Structure factors: contains datablocks I. DOI: crystallographic information; 3D view; checkCIF report Additional supplementary materials:"} +{"text": "We developed a method that allows the detection of circulating carcinoma cells in the blood of cancer patients. Circulating epithelial cells are harvested from peripheral blood mononuclear cells by immunomagnetic separation using BerEP4-coated beads. A telomeric repeat amplification protocol (TRAP)-ELISA is then used to measure telomerase in harvested epithelial cells. This method is specific and sensitive as demonstrated by experiments using BerEP4-positive and negative cell lines. Whereas we never found telomerase activity in harvested epithelial cells (HEC) samples from 30/30 healthy donors, we have detected telomerase activity in HEC from 11/15 (73%) patients with stage IIIB or IV non-small cell lung cancer (NSCLC) patients and from 8/11 (72%) stage C or D (Dukes classification) colon cancer patients. This non-invasive method could be of great value as a diagnostic or prognostic marker, or for monitoring cancer progression. \u00a9 2001 Cancer Research Campaign"} +{"text": "Pneumococcal surface protein A (PspA) elicits protection in mice against fatal bacteremia and sepsis caused by genetically diverse pneumococci and protects against carriage and lung infection. We determined the PspA families of invasive isolates of Streptococcus pneumoniae recovered from Colombian children <5 years of age. That 97.5% of Colombian isolates belong to PspA families 1 and 2 supports the hypothesis that a human PspA vaccine covering a few PspA families could be broadly effective."} +{"text": "IRG) play an important role in defense against intracellular pathogens. One member of this gene family in humans, IRGM, has been recently implicated as a risk factor for Crohn's disease. We analyzed the detailed structure of this gene family among primates and showed that most of the IRG gene cluster was deleted early in primate evolution, after the divergence of the anthropoids from prosimians ( about 50 million years ago). Comparative sequence analysis of New World and Old World monkey species shows that the single-copy IRGM gene became pseudogenized as a result of an Alu retrotransposition event in the anthropoid common ancestor that disrupted the open reading frame (ORF). We find that the ORF was reestablished as a part of a polymorphic stop codon in the common ancestor of humans and great apes. Expression analysis suggests that this change occurred in conjunction with the insertion of an endogenous retrovirus, which altered the transcription initiation, splicing, and expression profile of IRGM. These data argue that the gene became pseudogenized and was then resurrected through a series of complex structural events and suggest remarkable functional plasticity where alleles experience diverse evolutionary pressures over time. Such dynamism in structure and evolution may be critical for a gene family locked in an arms race with an ever-changing repertoire of intracellular parasites.Immunity-related GTPases ( IRG gene family plays an important role in defense against intracellular bacteria, and genome-wide association studies have implicated structural variants of the single-copy human IRGM locus as a risk factor for Crohn's disease. We reconstruct the evolutionary history of this region among primates and show that the ancestral tandem gene family contracted to a single pseudogene within the ancestral lineage of apes and monkeys. Phylogenetic analyses support a model where the gene has been \u201cdead\u201d for at least 25 million years of human primate evolution but whose ORF became restored in all human and great ape lineages. We suggest that the rebirth or restoration of the gene coincided with the insertion of an endogenous retrovirus, which now serves as the functional promoter driving human gene expression. We suggest that either the gene is not functional in humans or this represents one of the first documented examples of gene death and rebirth.The IRG), a family of genes induced by interferons, are one of the strongest resistance systems to intracellular pathogens IRGM gene has been shown to have a role in the autophagy-targeted destruction of Mycobacterium bovis BCGIRGM haplotypes associate with increased risk for Crohn's disease IRG gene family exists as multiple copies (3\u201321) in most mammalian species but has been reduced to two copies, IRGC and a truncated gene IRGM, in humans IRG genes except IRGC are organized in tandem gene clusters mapping to mouse chromosomes 11 and 18 (both syntenic to human chromosome 5) IRGM copy and IRGC in human IRG gene family in multiple nonhuman primate species in order to reconstruct the evolutionary history of this locus.Immunity Related GTPases (Microcebus murinus and Lemur catta) confirmed the mammalian archetypical organization with three IRGM paralogs in each species preserves a complete ORF based on the mouse model and shows the greatest homology to mouse Irgm1. The ORF encodes a putative 47 kD protein including a classical N-terminal region as well as classical motifs at the end of the carboxyl-terminus associated with most functional murine IRGM loci IRGM8, is likely a pseudogene because of a mutation generating a stop codon within the G domain and a frameshift mutation at the C terminus. The third mouse lemur gene, IRGM7, is atypical because it has substitutions in the G domain that disrupt the G1 motif that interacts with the nucleotide phosphates and is highly conserved in P-loop GTPases We next compared the structure of the c repeat immediately after the splicing acceptor that disrupts the ORF of the sole remaining IRGM gene suggested that the frameshift mutations were fixed likely as a result of an Alu repeat integration event that disrupted the ORF of the gene in the anthropoid ancestor could no longer be produced. These data strongly suggest that ERV9 integration significantly reshaped the expression and splicing pattern of IRGM in the common ancestor of humans and apes , chimpanzee (Ptr), gorilla (Ggo) and orangutan (Ppy)); Group 2 consists of species that carry a single copy of IRGM but lack the ERV9 element (Macaque (Rh), baboon (Pha) and marmoset (Cja)); while Group 3 was formed by species (dog and mouse lemur) that had multiple copies in a tandem orientation and Group 3 (\u03c9\u200a=\u200a0.3866) with an intermediate value for Group 1 (\u03c9\u200a=\u200a0.6073). Group 3 was found to be under constrained evolution (\u03c9\u200a=\u200a0.3866) and it was significantly different (P\u200a=\u200a6.09E\u221212) from a model of neutral evolution. In contrast, Group 1 and 2 gene evolutions were indistinguishable from a model of neutral evolution but was resurrected \u223c20 million years ago in the common ancestor humans and apes , gorilla (Gorilla Gorilla), orangutan (Pongo pygmaeus), rhesus macaque (Macaca mulatta), marmoset , baboon (Papio hamadryas), and Gray Mouse Lemur (Microcebus murinus) from NCBI Trace Archive (http://www.ncbi.nlm.nih.gov/Traces/trace.cgi?) and constructed local sequence assemblies using PHRAP (http://www.phrap.org). We sequenced and confirmed the IRGM genome organization based on DNA samples from four different New World monkey species and from eleven different Old World monkey species. We also resequenced the IRGM gene in unrelated macaques (n\u200a=\u200a5), baboons (n\u200a=\u200a5), orangutans (n\u200a=\u200a12) and gibbons (n\u200a=\u200a7). For Microcebus murinus with multiple copies of IRGM, we first isolated large-insert BAC clones, subcloned and sequenced PCR amplicons corresponding to the different copies.We retrieved whole genome shotgun sequence of the Homo sapiens), rhesus macaque (Macaca mulatta), marmoset and lemur (Lemur catta). FISH was performed using either human IRGM probe WIBR2-3607H18 or lemur IRGM BAC DNA LB2-61D22, LB2-77B23 and LB-61A22, directly labeled by nick translation with Cy3-dUTP (Perkin-Elmer). Lemur BAC probes were obtained by library hybridization screening of a L. catta genomic library (CHORI Resources: LBNL-2 Lemur BAC Library [http://gsd.jgi-psf.org/cheng/LB2]).Metaphase spreads were obtained from lymphoblast or fibroblast cell lines from human (IRGM transcript was obtained by 5\u2032RACE PCR followed by subcloning (PGEM-T easy) and sequencing (EU742619). RT-PCR experiments were performed using cDNA synthesized from mRNA extracted from total RNA . Total RNA was obtained from tissues isolated from chimpanzee, rhesus macaque, marmoset and human. IRGM splice variants were detected by a quantitative PCR assay using the LightCycler SYBR Green System (Roche) with primers IRGM (b)-(c)-(d) and IRGM all primers (Full-length human primers . TranscrN/dS) were performed by maximum likelihood using PAML We generated multiple sequence alignments using Clustal-WFigure S1IRGM proteins. Protein sequence alignment of primate, dog and mouse IRGM shows close homology in N-terminal GTPase binding domain (G domain). Canonical GTPase motifs are indicated by red boxes. The sequences are edited to maintain the open reading frame of Cja, Rh, and Pph IRGM, which are considered to be pseudogenes (names are indicated in red color). Species names are indicated as: Hs (Homo sapiens), Ptr (Pan trogylodytes), Ggo (Gorilla gorilla), Ppy (Pongo pygmaeus), Rh (Rhesus macaque-Macaca mulatta), Cja , Pph (Papio hamadryas), IRGM7, IRGM8, IRGM9 Mmu (Microcebus murinus), IRGM4, IRGM5, IRGM6 (Dog IRGM GMS type GTPases), IRGM1, IRGM2, IRGM3 (Mouse IRGM GMS type GTPases).Amino acid alignment of the (0.09 MB PDF)Click here for additional data file.Figure S2IRGM Alu repeat integration region. Blue highlighted sequence denotes the canonical splicing acceptor (based on murine gene model) with the red underlined sequence indicating the position of polypyrimidine tract. Green highlighted sequences correspond to the IRGM ORF. Alu integration site is indicated as red box (292 bp). Translation start site with preferred Kozak consensus sequence for Human IRGM is indicated as a green arrow. Stop codons in the ORF are indicated as red triangles.Alignment of the (0.09 MB PDF)Click here for additional data file.Figure S3IRGM. Phylogenetic reconstruction of IRGM related genes in different primate, dog and mouse species using the NJ method. Species names are indicated as: Mouse (Mus musculus domesticus), Dog (Canis familiaris), Gray mouse lemur (Microcebus murinus), Sbo (Saimiri boliviensis), Cge Marmoset , Cmo , Ppi (Pithecia pithecia), Mar (Macaca arctoides), Mni (Macaca nigra), Mmu Rhesus macaque (Macaca mulatta), Mfa (Macaca fascicularis), Pan (Papio hamadryas anubis), Pha Baboon (Papio hamadryas), Cce (Cercopithecus cephus), Cae (Cercopithecus aethiops), Pcr (Presbytis cristata), Cpo (Colobus polykomos), Cgu (Colobus guereza), Hga Gibbon (Hylobates gabriellae), Ppy Orangutan (Pongo pygmaeus), Ggo Gorilla (Gorilla gorilla), Ptr Chimpanzee (Pan troglodytes) and Hs Human (Homo sapiens). Shared stop codons for New World and Old World monkeys are highlighted in purple and blue respectively. Pseudogenes are highlighted in red.Phylogeny of (0.47 MB PDF)Click here for additional data file.Figure S4IRGM ERV9 region in . Red highlighted sequence denotes the ERV9 element. Yellow and green highlighted sequences correspond to the AluSc element and the IRGM ORF. Intron sequence is not included in this alignment indicated as red box (489 bp). Transcription start site (+1) indicated as green box. Stop codons in open reading frame are indicated as red triangles. Note the presence of a marmoset insertion sequence: (TAATGATAATTTCTAATCACTGCAAGAATCACATCACCTTCTTTGAATCAATCTCAAATACCTGGCCTGGTGGGAGCCAGGTTCTGCTCTTCTTCAAGG).Alignment of the (0.11 MB PDF)Click here for additional data file.Figure S5IRGM mRNA expression levels. A) A schematic summarizing the location of a sequenced structural polymorphism with respect to the IRGM gene ). The figure shows the relative fold expression of GM18507 (I/I), GM18555 (D/D) and GM15510 (I/D). C) Relative fold expression of IRGM (B) detected by real-time PCR. The figure shows a two-fold expression difference between a lymphoblastoid cell line homozygous for the 20.1 kb insertion GM18507 (I/I) and cell line homozygous for the deletion GM18555 (D/D).Structural variation and gene see . B) Rela(1.38 MB PDF)Click here for additional data file.Figure S6IRGM locus. A) A miropeats alignment comparing the human chromosome 5 reference sequence to a sequence from an alternate haplotype . The alignment depicts a 20.1 kb deletion region 5\u2032 upstream of the human IRGM. Arrow indicates the transcription start point within the ERV9 retroviral element. Green box represents IRGM open reading frame; red boxes indicate exons for adjacent MST150 gene. B) Array comparative genomic hybridization (aCGH) results for nine human DNA samples (four African and four non-African) against a reference genome DNA sample (NA15510). The analysis confirms a 20.1 kb deletion polymorphism (indicated as red dotted line) located at a distance of 2.82 kb 5\u2032 to the IRGM transcription start site. The individual NA15510 is hemizygous (one copy) and is used as the reference in these experiments.Structural polymorphism 5\u2032 upstream of the (0.42 MB PDF)Click here for additional data file.Text S1IRGM gene.Supplementary note: Death and resurrection of the human (0.61 MB PDF)Click here for additional data file."} +{"text": "We have examined the biological properties of CEA6, a human carcinoembryonic antigen (CEA)-specific single-chain Fv (scFv) isolated by phage display, and five related clones derived by affinity maturation and selected for improved off-rate (Koff). All clones bind strongly and specifically to CEA-positive human tumours by immunocytochemistry and show negligible cross-reactivity with normal colon. Flow cytometry of scFv on human liver cells indicates a shift in fine epitope specificity resulting from mutagenesis. All monomeric scFv have been radioiodinated, retaining effectively full binding activity. A single intravenous injection into nude mice bearing human colon tumour xenografts confirms tumour targeting in all cases. As reported in other studies, the kidney is the main route of elimination of scFv at early time points. Tumour binding of the parental antibody CEA6 consistently gives the highest tumour-blood ratios at 24 h (mean 16:1). Clone TO6D11, which has a sevenfold reduced Koff relative to CEA6, showed no difference in tumour uptake at 24 h but persisted at the tumour site for longer than CEA6. This study demonstrates a possible correlation between binding affinity and tumour residence time when examined in this model."} +{"text": "A recent report has provided strong evidence for a major prostate cancer susceptibility locus (HPC1) on chromosome 1q24-25 . Most inherited cancer susceptibility genes function as tumour-suppressor genes (TSGs). Allelic loss or imbalance in tumour tissue is often the hallmark of a TSG. Studies of allelic loss have not previously implicated the chromosomal region 1q24-25 in prostate cancer. However, analysis of tumour DNA from cases in prostate cancer families has not been reported. In this study, we have evaluated DNA from tissue obtained from small families [3-5 affected members (n = 17)], sibling pairs (n = 15) and sporadic (n = 40) prostate tumours using the three markers from Smith et al (1996) that defined the maximum multipoint linkage lod score. Although widely spaced (12-50 cM), each marker showed evidence of allelic imbalance in only approximately 7.5% of informative tumours. There was no difference between the familial and sporadic cases. We conclude that the incidence of allelic imbalance at HPC1 is low in both sporadic tumours and small prostate cancer families. In this group of patients, HPC1 is unlikely to be acting as a TSG in the development of prostate cancer."} +{"text": "Univariate logistic regression models revealed a significant influence of MCP-1 serum levels on the odds of presenting with primary ovarian cancer versus benign cysts and versus healthy women respectively . In a multivariate logistic regression model considering MCP-1 and CA 125 serum levels simultaneously, both MCP-1 and CA 125 revealed statistical significance on the odds of presenting with primary ovarian cancer versus benign cysts . In ovarian cancer patients, MCP-1 serum levels showed a statistically significant correlation with histological grade and age at the time of diagnosis . Elevated MCP-1 serum levels prior to therapy were not associated with disease-free and overall survival . In summary these data indicate that MCP-1 might play a functional role in the natural history of ovarian cancer and might serve as differentiation marker between benign ovarian cysts and ovarian cancer, providing additional information to the established tumour marker CA 125. \u00a9 1999 Cancer Research CampaignThe chemokine monocyte chemoattractant protein (MCP)-1 is an important mediator of monocyte infiltration in various solid tumours of epithelial origin. The aim of the present study was to evaluate the role of MCP-1 in the natural history of ovarian cancer and to determine its value as differentiation marker and prognostic marker regarding disease free and overall survival. This retrospective study comprises 86 patients with ovarian cancer, 48 with primary ovarian cancer and 38 with recurrent ovarian cancer, 67 patients with benign ovarian cysts and 42 healthy women. Median serum levels in patients with primary ovarian cancer, recurrent ovarian cancer, benign ovarian cysts and in healthy women were 535.6 (range 129.6\u20131200) pg ml"} +{"text": "Aims To asses whether clinically severe insulinresistance and poor metabolic control in patients withtype II diabetes are associated with aberrantexpression or function of the p21ras pathway. Methods We examined the expression and functionof the p21ras pathway in resting and activated PBMCfrom 10 insulin treated patients with type II diabetescharacterized by high insulin requirements and poormetabolic control (IR group) and 10 age and sexmatched well controlled patients treated by dietalone or oral hypoglycemic medications (WC group). Results Levels of p21ras and its regulatoryelements: p21rasGAP and hSOS1, were comparablein the two groups. The induced activities of p21rasand its associated down-stream regulatory enzymeMAP-kinase following TPA stimulation were alsocomparable in the IR and WC patients. Conclusions Taken together, these data indicate thatclinically significant severe insulin resistance doesnot modify the expression, regulation and activationof p21ras pathway in PBMC of patients with type IIdiabetes."} +{"text": "The frequency of DNA aneuploidy was investigated by flow cytometry in 156 colorectal adenomas including 56 associated with 36 synchronous adenocarcinomas. Nine of 156 adenomas (6%) were DNA aneuploid. DNA aneuploidy correlated with increasing size (P less than 0.005) and histopathological type P less than 0.05) but not with dysplasia. Adenomas in associated with a synchronous adenocarcinoma did not have an increased incidence of DNA aneuploidy. Adenocarcinomas found in association with adenomas tend to have a lower incidence of DNA aneuploidy then the generality of colorectal cancers."} +{"text": "Automatic semantic role labeling (SRL) is a natural language processing (NLP) technique that maps sentences to semantic representations. This technique has been widely studied in the recent years, but mostly with data in newswire domains. Here, we report on a SRL model for identifying the semantic roles of biomedical predicates describing protein transport in GeneRIFs \u2013 manually curated sentences focusing on gene functions. To avoid the computational cost of syntactic parsing, and because the boundaries of our protein transport roles often did not match up with syntactic phrase boundaries, we approached this problem with a word-chunking paradigm and trained support vector machine classifiers to classify words as being at the beginning, inside or outside of a protein transport role.We collected a set of 837 GeneRIFs describing movements of proteins between cellular components, whose predicates were annotated for the semantic roles AGENT, PATIENT, ORIGIN and DESTINATION. We trained these models with the features of previous word-chunking models, features adapted from phrase-chunking models, and features derived from an analysis of our data. Our models were able to label protein transport semantic roles with 87.6% precision and 79.0% recall when using manually annotated protein boundaries, and 87.0% precision and 74.5% recall when using automatically identified ones.We successfully adapted the word-chunking classification paradigm to semantic role labeling, applying it to a new domain with predicates completely absent from any previous studies. By combining the traditional word and phrasal role labeling features with biomedical features like protein boundaries and MEDPOST part of speech tags, we were able to address the challenges posed by the new domain data and subsequently build robust models that achieved F-measures as high as 83.1. This system for extracting protein transport information from GeneRIFs performs well even with proteins identified automatically, and is therefore more robust than the rule-based methods previously used to extract protein transport roles. Automatic semantic role labeling (SRL) is a natural language processing (NLP) technique that maps sentences to semantic representations, which can be useful for many NLP tasks (e.g. information extraction). With the advent of resources like FrameNet and PropAs a variety of research groups have reported success on these corpora, recent work has turned to transferring these results to different kinds of predicates and different genres of text. In this article, we show that automatic semantic role labeling can be transferred to the biomedical domain. Our goal is to accept as input sentences describing biological processes and infer structures like the following:PATIENT Bax] [PREDICATE translocation] from the [ORIGIN cytosol] to [DESTINATION mitochondria] leads to the subsequent formation....(1) workers [PREDICATE(VERB) have] asbestos-related diseases, [PREDICATE(VERB) including] three with recently [PREDICATE(VERB) diagnosed] cancer.(2) Four of the five of up to 44 C-terminal amino acids from the putatively cytoplasmic C-terminal hydrophilic domain left transport function [PREDICATE(ADJ) unimpaired], but [PREDICATE(NOUN) deletion] of the adjacent STAS (sulfate transporter anti-sigma factor antagonist) domain [PREDICATE(VERB) abolished] function.(3) [PREDICATE translocation] from the [ORIGIN cytosol] to the [DESTINATION plasma membrane].(4) IRS-3 expression blocked glucose/IGF-1 induced translocation(5) [PATIENT GLUT-4] translocation(6) [DESTINATION nuclear] translocation(7) [PREDICATE translocation]](8) regulate its [ORIGIN nuclear] [PREDICATE export] during glucose deprivation(10) Tryptophan 521 and serine 667 residues of [PREDICATE translocation] to the [DESTINATION nucleus](11) , overexpressed in prostate cancer, [PREDICATE shuttles] between the cytoplasm and the nucleus.(12) This [DESTINATION1 nuclear] [PREDICATE1 import] and inhibiting [PATIENT2 BRCA1] [ORIGIN2 nuclear] [PREDICATE2 export](13) BARD1 induces BRCA1 intranuclear foci formation by increasing RING-dependent [DESTINATION1 nuclear] [PREDICATE1 import] and inhibiting [PATIENT2 BRCA1] [ORIGIN2 nuclear] [PREDICATE2 export](14) BARD1 induces BRCA1 intranuclear foci formation by increasing RING-dependent requires [PREDICATE translocation]...(15) These results suggest that [PATIENT eNOS] [PREDICATE translocation]...(16) ... involved in folding and [PREDICATE translocation]...(17) ... for ERdj5 in [PATIENT Tir] secretion and [PREDICATE translocation]...(18) ... for efficient correlating with its microtubule- and microfilament-mediated [PREDICATE translocation](19) a rapid activation of the [PREDICATE translocation] from the [ORIGIN cytosol] to [DESTINATION mitochondria] leads to the subsequent formation. ...(1) [PREDICATE(NOUN) Truncation] of up to 44 C-terminal amino acids from the putatively cytoplasmic C-terminal hydrophilic domain left transport function [PREDICATE(ADJ) unimpaired], but [PREDICATE(NOUN) deletion] of the adjacent STAS (sulfate transporter anti-sigma factor antagonist) domain [PREDICATE(VERB) abolished] function.(3) [PATIENT IRS-2] [PREDICATE translocation] from the [ORIGIN cytosol] to the [DESTINATION plasma membrane].(4) IRS-3 expression blocked glucose/IGF-1 induced [PATIENT p53] stability via the inhibition of its [ORIGIN nuclear] [PREDICATE export] mechanism.(9) Serine 392 exerts important effects upon [PATIENT Daxx] regulate its [ORIGIN nuclear] [PREDICATE export] during glucose deprivation(10) Tryptophan 521 and serine 667 residues of [PATIENT Insulin receptor substrate 1] [PREDICATE translocation] to the [DESTINATION nucleus](11) [PATIENT protein], overexpressed in prostate cancer, [PREDICATE shuttles] between the cytoplasm and the nucleus.(12) This ["} +{"text": "The potential role of endogenous sex hormones in regulating hypothalamo\u2013pituitary\u2013adrenal (HPA) axis function was investigated after a single injection of endotoxin in adult (8 week old) BALB/c mice of both sexes. The effect of LPS on plasma ACTH, corticosterone (B), testosterone and oestradiol (E) levels and on anterior pituitary (AP) ACTH and adrenal B contents at different times after treatment was studied. The results indicate that: (a) basal B but not ACTH plasma levels were significantly higher in female than in male mice; (b) LPS significantly increased both ACTH and B plasma levels over the baseline 2 h after injection, both hormone levels being higher in female than in male mice; (c) although plasma ACTH concentrations recovered the basal value at 72 h after LPS in animals of both sexes, plasma B levels returned to the baseline only at 120 h after treatment; (d) E plasma levels significantly increased 2 h after LPS and returned to the baseline at 72 h post-treatment, in both sexes; (e) at 2 h after LPS, testosterone plasma levels significantly decreased in male mice and increased in female mice, recovering the baseline level at 120 and 72 h after LPS, respectively; (f) AP ACTH content was similar in both sexes in basal condition and it was significantly diminished 72 h post-treatment without sex difference; whereas AP ACTH returned to basal content 120 h after LPS in males, it remained significantly decreased in females; (g) basal adrenal B content was higher in female than in male mice, and it significantly increased in both sexes 2 h post-LPS, maintaining this sex difference. Whereas adrenal B returned to basal content 72 h after treatment in male mice, it remained significantly enhanced up to 120 h post-LPS in female animals. The data demonstrate the existence of a clear sexual dimorphism in basal condition and during the acute phase response as well as in the recovery of the HPA axis function shortly after infection."} +{"text": "Cerebral metastases of cutaneous melanoma carry a very poor prognosis. We report our experience of 31 patients who presented with cerebral metastasis of cutaneous melanoma in a 5-year period between mid-1991 and mid-1996. Cerebral metastases were diagnosed on computerized tomography (CT) scan after patients became symptomatic. The overall median survival in our series was 4 months. Seventeen patients (55%) received treatment with radiotherapy and dexamethasone with resolution of their symptoms, although median survival remained at 4 months. Six patients (19%) had surgery followed by whole brain radiotherapy, with median survival of 5 months. The remaining eight patients received dexamethasone alone. Data from patients surviving less than 2 months and over 6 months suggest that the poor prognostic factors are the presence of more than one cerebral metastasis and additional extracranial metastases."} +{"text": "TGF\u03b2RII gene. The aim of our study was to evaluate the significance of simple tests performed on tumours to select appropriate candidates for germline mutational analysis. We studied three groups of patients, HNPCC kindreds fulfilling the International Collaborative Group (ICG) criteria (n = 10), families in which at least one of the criteria was not satisfied (n = 7) and sporadic colorectal cancer (CRC) diagnosed before the age of 50 (n = 17). We searched for microsatellite instability (MSI), presence of hMSH2 and hMLH1 germline mutations, expression of hMSH2, hMLH1 and p53 proteins in tumoural tissue samples by immunostaining. Fifteen out of 17 (88%) of HNPCC and incomplete HNPCC cases were MSI and eight pathogenic germline mutations in hMSH2 or hMLH1 were detected in these two groups (53%). All the 17 early-onset sporadic cases were MSS and no germline mutations were detected among the seven investigated cases. Thirteen out of 15 (81%) familial cases were MSI and p53 protein-negative, whereas 13/14 (93%) sporadic cases were MSS and strongly p53 protein-positive. This extensive molecular investigation shows that simple tests such as MS study combined with hMSH2 and hMLH1 protein immunostaining performed on tumoural tissues may provide valuable information to distinguish between familial, and probably hereditary, and sporadic CRC cases. \u00a9 2000 Cancer Research CampaignThe genetic abnormalities underlying hereditary non-polyposis colorectal cancer (HNPCC) are germline mutations in one of five DNA mismatch repair genes or in the"} +{"text": "Hlnd[RuInd2Cl4] will react with apotransferrin only in the presence of bicarbonate, thisanion dictating the kinetic and mechanistic characteristics of protein-binding.Circular dichroism studies had previously indicated that binding of both Ru(III) complexes occursaround the unoccupied iron(III) binding sites; this result is now confirmed by preliminary X-raydata of Hlnd[RuInd2Cl4] and Hlm[RuIm2Cl4] bound to apolactoferrin, a related iron protein. Thecrystallograhic data reveals that binding of both complexes takes place at histidine residues, andthat the ligand (indazole) remains bound in the case of Hlnd[RuInd2Cl4].The interaction of two ruthenium(III) complexes exhibiting high anticancer activity - namely"} +{"text": "BRCA1 mutations have an increased risk for colorectal cancer (CRC). To gain insight into this issue, 225 unselected Ashkenazi Jewish CRC patients were tested for the presence of the three common Jewish BRCA1/2 germline mutations: 185delAG and 5382insC (BRCA1) and 6174delT (BRCA2). A total of four carriers was found . This frequency is similar to the estimated normal Ashkenazi population frequency, thus suggesting that these specific mutations do not contribute to CRC predisposition.\u00a9 2001 Cancer Research Campaign http://www.bjcancer.comIt is presently unclear whether carriers of"} +{"text": "This dose-intensified CMF schedule was accompanied by enhanced haematological toxicity with clinical sequelae, namely fever, intravenous antibiotics and red blood cell transfusions, but allows a high dose intensity in a majority of patients. \u00a9 2000 Cancer Research CampaignOur aim was to study the feasibility of an intensified intravenous CMF schedule with the aim to escalate dose intensity (DI). Twenty-three premenopausal breast cancer patients received 6 cycles of adjuvant CMF intravenously on days 1 and 8 every 3 weeks and granulocyte colony-stimulating factor days 9\u201318. Endpoints were DI and toxicity. Twenty-one out of 23 patients (91%) received the projected total dose and reached \u2265 85% of the projected DI. Compared to \u2018classical\u2019 CMF, all patients reached \u2265 111% DI. Nine patients received the planned schedule without delay. Thirteen patients (57%) were treated for infection and four patients (17%) were hospitalized for febrile neutropenia. Twelve patients received red blood cell transfusions (52%). Radiation therapy ("} +{"text": "A reduction in tumour yield was apparent when progesterone administration was begun 25 days before feeding 7,12-dimethylbenz(a)anthracene (DMBA). This effect was most obvious when the duration of hormone administration was brief. Continuation of progesterone for some time after feeding DMBA caused a progressive diminution of the inhibitory effect, and 135 days of continuous hormone treatment entirely abolished the effects of 25 days pretreatment with the hormone.In contrast, when progesterone injections were begun 2 days after feeding DMBA, there was a trend towards enhancement of tumour yield. Continuous hormone administration appeared more effective than shorter treatment regimens."} +{"text": "A cyclic compound of composition {[(en)Pt]4}4+ (UH=monoanion of unsubstituted uracil) is presented and theanalogy with organic calix-[4]-arenes is pointed out. (iii) Cyclic nucleobase complexes from trans-a2Pt(II). Possible ways for the preparation of macrocyclic nucleobase complexes containing trans-a2Pt(II) linkages are outlined and precursors and intermediates are presented.Of all properties of metal nucleobase complexes, formation of multinuclear species appears to bean outstanding feature. After a brief introduction into well known polymeric metal nucleobasecomplexes, three aspects recently Studied in our laboratory will be dealt with in more detail: (i) Heteronuclearcomplexes derived from trans-[(amine)"} +{"text": "We analysed aromatase gene expression and its regulation in seven cases of endometrioid endometrial carcinoma. Immunohistochemistry revealed the presence of strong aromatase immunoreactivity in the stromal cells of carcinoma in five out of seven cases. A polymerase chain reaction after reverse transcription (RT PCR) revealed varying levels of aromatase transcripts (0.1-27.0 amol ng(-1) total mRNA) in five cases. The alternative use of multiple exons 1 was also examined by identifying various human aromatase transcripts specific for exons 1 in RT-PCR products. Gonadal type or exon 1d was primarily used in three cases in which aromatase overexpression was not detected. The two cases in which fibroblasts type or exon 1b was used with other exons 1 as minor transcripts demonstrated aromatase overexpression in immunohistochemistry and RT PCR analysis. Further studies are required, but alternative splicing as well as use of multiple exons 1 transcripts may result in increased aromatase expression in stromal cells observed in endometrial carcinoma."} +{"text": "PTEN is a novel tumour-suppressor gene located on chromosomal band 10q23.3. This region displays frequent loss of heterozygosity (LOH) in a variety of human neoplasms including breast carcinomas. The detection of PTEN mutations in Cowden disease and in breast carcinoma cell lines suggests that PTEN may be involved in mammary carcinogenesis. We here report a mutational analysis of tumour specimens from 103 primary breast carcinomas and constitutive DNA from 25 breast cancer families. The entire coding region of PTEN was screened by single-strand conformation polymorphism (SSCP) analysis and direct sequencing using intron-based primers. No germline mutations could be identified in the breast cancer families and only one sporadic carcinoma carried a PTEN mutation at one allele. In addition, all sporadic tumours were analysed for homozygous deletions by differential polymerase chain reaction (PCR) and for allelic loss using the microsatellite markers D10S215, D10S564 and D10S573. No homozygous deletions were detected and only 10 out of 94 informative tumours showed allelic loss in the PTEN region. These results suggest that PTEN does not play a major role in breast cancer formation. 1999 Cancer Research Campaign"} +{"text": "The geometric and subcellular organization of axon arbors distributes and regulates electrical signaling in neurons and networks, but the underlying mechanisms have remained elusive. In rodent cerebellar cortex, stellate interneurons elaborate characteristic axon arbors that selectively innervate Purkinje cell dendrites and likely regulate dendritic integration. We used GFP BAC transgenic reporter mice to examine the cellular processes and molecular mechanisms underlying the development of stellate cell axons and their innervation pattern. We show that stellate axons are organized and guided towards Purkinje cell dendrites by an intermediate scaffold of Bergmann glial (BG) fibers. The L1 family immunoglobulin protein Close Homologue of L1 (CHL1) is localized to apical BG fibers and stellate cells during the development of stellate axon arbors. In the absence of CHL1, stellate axons deviate from BG fibers and show aberrant branching and orientation. Furthermore, synapse formation between aberrant stellate axons and Purkinje dendrites is reduced and cannot be maintained, leading to progressive atrophy of axon terminals. These results establish BG fibers as a guiding scaffold and CHL1 a molecular signal in the organization of stellate axon arbors and in directing their dendritic innervation. Large principal neurons in vertebrate neural circuits often consist of distinct anatomical and physiological compartments, which allow distributed and compartmentalized signaling and greatly increase the computational power of single neurons. Superimposed upon this intrinsic compartmental architecture is the subcellular organization of synaptic inputs, which exert local control over the biophysical properties and differentially regulate the input, integration, and output of principal neurons. In the cerebellar cortex, Purkinje neurons are innervated by GABA inhibitory synapses from the stellate and basket cells at dendrites and soma-axon initial (AIS) segments, respectively. Previous studies have shown that an L1 family immunoglobulin cell adhesion molecule (neurofascin186) is distributed as a subcellular gradient and directs basket cell axons to innervate Purkinje cell AIS. Here, we examine the mechanisms underlying the innervation of Purkinje cell dendrites by stellate axons. We found that stellate axons are organized into characteristic trajectories and guided towards Purkinje dendrites by an intermediate scaffold of astroglia\u2014the Bergmann glial (BG) fibers. Another member of L1 family, Close Homologue of L1 (CHL1), is localized to BG fibers and stellate cells, and contributes to the organization of stellate axons along BG fibers and to the innervation of Purkinje cell dendrites. Subcellular synapse organization regulates the input, integration, and output of target neurons. An astroglial scaffold and an L1 family cell adhesion molecule contribute to dendritic innervation by GABA inhibitory synapses. Neurons are often characterized by striking polarity and extensive subcellular specialization. For example, large principal neurons in many vertebrate neural circuits consist of distinct anatomical and physiological compartments , which aA prime example of subcellular synapse organization is the Purkinje neurons of the cerebellum. The cerebellar cortex is organized as a near lattice-like circuit architecture along the two axes of the cerebellar lobules, the translobular and parlobular planes . At the Stellate cells mainly occupy the upper half of the molecular layer (ML) and are the only cell type of the upper third of the ML. Like the basket cells, stellate cells extend their axons within the translobular plane of the cerebellar cortex . AlthougBergmann glia (BG) cells are highly polarized astrocytes, whose radial fibers dominate the cerebellar cortex ,14,15. DThe L1 family immunoglobulin cell adhesion molecules (L1CAMs) have been implicated in axon growth, guidance , and theTo investigate the cellular mechanisms underlying development of the stellate axon arbor and innervation pattern, it is necessary to visualize stellate axons together with their postsynaptic targets at high resolution and during the developmental process. We generated several lines of bacterial artificial chromosome (BAC) transgenic reporter mice to achieve such visualization. The calcium binding protein parvalbumin (Pv) is normally expressed in all Purkinje, basket, and stellate neurons in the postnatal cerebellar cortex. In our Pv-GFP BAC transgenic mice A, differStellate and basket cells occupy mainly the upper or lower half of the ML, respectively. Consistent with previous Golgi studies , GFP labTo better understand the developmental process by which stellate axons innervate Purkinje dendrites, we used our reporter mice expressing GFP from the GAD67 promoter elements . These Besides Purkinje dendrites, an equally prominent cellular component of the cerebellar cortex are the BG fibers FiguresF and S1.The vertical bias of the orientation of stellate axon collaterals prompted us to examine their relationship with BG fibers during the development of dendritic innervation. As expected, when stellate cell precursors were migrating across the PCL in the second postnatal week, GFAP-positive BG fibers were prominent throughout the ML , indicating that the organization of GAD65 along BG fibers was not due to chance. This analysis is likely an underestimate of GAD65\u2013BG association since GFAP antibodies did not label well the finer BG processes littermates, \u2212/\u2212L1 mice, and \u2212/NrCAM\u2212 mice of GAD65 in \u2212/\u2212CHL1 mice compared to their WT littermates mice. In the ML of mature WT B20 mice (P44), stellate axons display complex arbors with characteristic orientations A; a majo\u2212/\u2212PV-GFP(B20)::CHL1 littermates, most stellate axons still were able to develop fairly complex arbors at this age, but appeared thinner, more wavy, with significantly altered orientation and trajectories . At P30, there was also a significant reduction in the density of symmetric synapses by approximately 40% (p < 0.03). On the other hand, basket axon synapses on Purkinje somata, parallel fiber synapses on dendritic spines, and climbing fiber synapses on dendritic shafts were all indistinguishable between P44 WT and \u2212/\u2212CHL1 mice . We also deleted CHL1 in Purkinje cells by breeding CHL1flx mice with the L7-Cre transgenic mice . A. A11]. AIn mature cerebellar cortex, each BG cell gives rise to several ascending BG fibers, which extend approximately 40\u201350 \u03bcm in the translobular plane and 15\u201320 \u03bcm in the parlobular plane ,15. InteA receptors that enwrap inhibitory synapses ) nor climbing fibers (CF [G\u2013K]) synapses showed any discernable defects in (8.53 MB TIF)Click here for additional data file.Figure S6\u2212/\u2212CHL1 mice extended their axons but failed to associate with the GFAP-labeled BG fibers (arrows).(A) At P16, stellate cells in (B and C) At more-mature ages (P20 and P40), stellate cell axons were still largely not associated with BG fibers. Note that at P40 (C), some of these stellate axons extended rather randomly, twisted, tangled, and even circled around (arrows). See (8.36 MB TIF)Click here for additional data file.Figure S7\u2212/\u2212CHL1 mice and expressed GAD65 .(A) At single\u2013basket cell resolution from PV-GFP (B20 mice), pinceau synapses (arrows) developed normally in \u2212/\u2212CHL1 mice (B2\u20133) as in WT mice ((B) Basket axons (green) grew along Purkinje proximal dendrite in WT mice H.\u2212/CHL1\u2212 (D) mice.(C and D) Ultrastructural analysis revealed similar basket synapses onto Purkinje soma in WT (C) and Bt, basket; Pc, Purkinje cell. Scale bars indicate 20 \u03bcm.(8.58 MB TIF)Click here for additional data file."} +{"text": "Preincubation with H2O2 (10\u03bcM) produced a significant decrease in PE (1mM)-induced contraction in untreated-diabeticbut not in control rats. Single treatment withinsulin counteracted this effect of H2O2 andalso reversed the increased contractile responseof diabetic aorta to PE, while vitamin A wasfound to be ineffective. H2O2 (10 \u03bcM) alsoinhibited ACh (1 mM)-stimulated endothelium-dependent relaxation two fold more in diabeticthan in control aorta. In the prevention ofH2O2-induced inhibition of vascular relaxationto ACh, vitamin A alone was markedly effectivewhile insulin alone was not. The combinationof vitamin A plus insulin removed theinhibitory action of H2O2 in diabetic aorta.Diabetic animals displayed an increased level of aorta thiobarbituric acid reactive substance(TBARS) in association with decreased levels ofplasma retinol and retinol-binding protein(RBP). Single treatment with insulin, in spite ofallowing recovery of normal growth rate andimproved glucose and retinol metabolism indiabetic rats, was unable to control TBARSproduction to the same extent as vitamin Aalone. Our findings suggest that the maintenanceof ACh-stimulated endothelium-dependentvasorelaxant tone in normal physiologicallevels depends largely on the prevention and/orinhibition of peroxidative stress, which isachieved by combined treatment with vitaminA plus insulin. The use of vitamin A togetherwith insulin provides a better metabolic controland more benefits than use of insulin alone inthe reduction of diabetes-induced vascularcomplications.A positive correlation has been establishedbetween increased oxidative stress and cardiovasculardiseases in diabetes mellitus. We evaluatedthe effects of single or combined treatmentswith vitamin A and insulin on vasomotoractivity, oxidative stress and retinol metabolismin 12-week streptozotocin diabetic rats. Thevasomotor activity was determined by measuringin"} +{"text": "The vascularisation of human primary colorectal carcinomas was studied immunohistochemically using the endothelial cell markers CD31 and factor VIII-related antigen. Tumour sections were systematically scanned at a magnification of x 100 to find areas of intense neovascularisation. Microvessel counts within these vascular 'hotspots' were performed at magnification x 250. Regions in which tumour cords were surrounded by a collagen IV-positive basement membrane were compared with those in which this was absent and with normal mucosa. CD31 appeared to be a more sensitive marker for endothelial cells than factor VIII-related antigen (mean 185 +/- 59 and 120 +/- 38 microvessels mm-2). Within individual tumour sections microvessel counts in vascular hotspots with highest vessel density correlated significantly with microvessel counts in vascular hotspots with second highest vessel density (P < 0.01). Microvessel counts in tumour areas where collagen IV-positive basement membrane were absent exceeded those in areas where it was present (factor of 1.7) and those in normal mucosa (factor of 1.6). The differences in vessel density between individual tumours and the low variability in vessel density within individual tumours using this quantification technique allow us to investigate the prognostic value of vessel density in areas of intense neovascularisation in human primary colorectal carcinomas."} +{"text": "We previously reported the efficacy of nonmyeloablative allogeneic transplantation in 2 HIV positive recipients, one of whom received retrovirus transduced hematopoietic stem cells to confer resistance to HIV. Here we report an assessment of retroviral integration sites (RISs) recovered out to 3 years post-transplantation. We identified 213 unique RISs from the patient's peripheral blood samples by linear amplification-mediated PCR (LAM-PCR). While vector integration patterns were similar to that previously reported, only 3.76% of RISs were common among early (up to 3 months) and late samples (beyond 1 year). Additionally, common integration sites were enriched among late samples . Three RISs were found near or within known oncogenes, but 2 were limited to early timepoints. Interestingly, an integration site near the MDS1 gene was detected in long-term follow-up samples; however, the overall contribution of MDS1 integrated clone remained stably low during follow-up. LMO2 gene in patients treated for severe combined immunodeficiency MDS1-EVI1, PRDM16, or SETBP1 integration containing myeloid cells observed in patients treated for chronic granulomatous disease using a similar approach Mds1/Evi1 gene MDS-EVI1 translocation 3\u223621 found in human acute myelogenous leukemia (AML) The establishment of safe and effective vector delivery systems for gene therapy applications appears achievable; however, the adverse events reported from recent gene therapy trials have lead to a reassessment of the risks associated with vector insertional mutagenesis TasI enzymes and TaiI enzymes were used , located in a region of frequent homozygous deletions in tumor samples ARL11), a genetic variant of which predisposes to familial cancerMDS1 .We found a total of 15 common integration sites . As the frequency of CISs increased in late phase populations, these results suggest that integrations at CISs dominate gene-modified long-term hematopoiesis. These observations after long term follow-up in a human gene therapy trial are in agreement with an emerging theory that vector integrations may serve as a tool to query genes involved in hematopoiesis in vivo We found total of 15 common integration sites (CISs) . None of the CISs were near or within known oncogenes. Only 14.9% (20 of 134 early RISs) were CISs, while 36.8% (32 of 87 late RISs) were CISs gene-therapy clinical trials could in part be explained by a lower infused cell dose along with the disease context MDS1 region occurred in a LT-HSC and though theoretically it might impact engraftment or survival of the LT-HSC, it did not result in an abnormal proliferation, clonal expansion, or oncogenesis in our patient. Recent reports In contrast, the clonal dominance of In summary, the retroviral integration pattern observed in our HIV gene therapy trial is similar to that previously observed in model systems and human clinical trials, yet several novel observations warrant emphasis. The pattern of contribution by genetically modified cells is distinct between the early and late phase post transplantation and emphasizes the importance of long-term studies to assess the risk of integrating vectors. Additionally, the enrichment for CISs in the late phase supports the concept that integrations in the LT-HSCs favors genes that may be involved in \u201cstemness\u201d www.clinicaltrials.gov (NCT 00005785), and are described here briefly. An HIV positive patient with treatment-related AML underwent nonmyeloablative allogeneic transplantation from an HLA-matched sibling. Half of the donor cells were genetically modified with a Moloney murine leukemia virus (MoMLV) based HIV resistance vector containing a transdominant negative mutant Rev (TdRev) 8 cells) or a control vector MoMLV based vector encoding gp91phox (4.04\u00d7108 cells). The transduction efficiency was estimated by PCR, with 80% efficiency achieved from the TdRev aliquot and 90% for the gp91phox aliquot 3 long-term while continuing on the same highly active antiretroviral therapy (HAART) regimen until her death from thrombotic thrombocytopenic purpura 3 years and 4 months post transplant. Vector-transduced cells remained detectable at low levels. DNA was isolated from patient blood sample as previously described phox and TdRev vectors in both lymphoid and myeloid cells at extended follow up The protocol was approved by the Institutional Scientific Review Committee and the Institutional Review Board of the National Heart, Lung, and Blood Institute, the Recombinant DNA Advisory Committee, and the Food and Drug Administration. All the study subjects gave written informed consent. The study design and outcome have been previously published To identify genomic-proviral integration sites, LAM\u2013PCR was performed as previously described MDS1, PCR was performed on 10 ng DNA using the primer set in MDS1 integrated clone's contributions over time after engraftment was determined by Real-time PCR analysis (QPCR) with Taqman probes as previously described To confirm the presence of an insertion near Table S1(0.04 MB DOC)Click here for additional data file.Table S2(0.13 MB DOC)Click here for additional data file.Table S3(0.09 MB DOC)Click here for additional data file.Table S4(0.04 MB DOC)Click here for additional data file."} +{"text": "In order to assess the blood flow patterns through human lung tumours, 20 patients received 400-750 MBq 99TcmHMPAO intravenously 10 min before single photon emission computed tomography (SPECT). Ratios of uptake in the whole tumour relative to normal lung ranged from 0.35 to 1.53 (mean 1.01) with eight tumours showing less uptake than normal lung and ten showing greater uptake. In one patient the tumour was not distinguishable from surrounding lung and in another a large pleural effusion prevented evaluation. Tumour: lung ratios for central tumour regions ranged from 0 to 1.83 (mean 0.80) with 13 showing lower uptake than normal lung and five showing greater uptake. Duplicate scans were performed in eight patients demonstrating satisfactory reproducibility. This technique provides a simple and reproducible method for the assessment of tumour blood flow."} +{"text": "The optimal use of mitoxantrone (NOV) in the high-dose range requires elucidation of its maximum tolerated dose with peripheral blood progenitor cell (PBPC) support and the time interval needed between drug administration and PBPC reinfusion in order to avoid graft toxicity. The aims of this study were: (1) to verify the feasibility and haematological toxicity of escalating NOV up to 90 mg m(-2) with PBPC support; and (2) to verify the safeness of a short (96 h) interval between NOV administration and PBPC reinfusion. Three cohorts of ten patients with breast cancer (BC) or non-Hodgkin's lymphoma (NHL) received escalating doses of NOV, 60, 75 and 90 mg m(-2) plus melphalan (L-PAM), 140-180 mg m(-2), with PBPC rescue 96 h after NOV. Haematological toxicity was evaluated daily (WHO criteria). NOV plasma pharmacokinetics was also evaluated, as well as NOV cytotoxicity against PBPCs. Haematological recovery was rapid and complete at each NOV dose level without statistically significant differences, and there were no major toxicities. NOV plasma concentrations at the time of PBPC reinfusion were below the toxicity threshold against haemopoietic progenitors. It is concluded that, when adequately supported with PBPCs, NOV can be escalated up to 90 mg m(-2) with acceptable haematological toxicity. PBPCs can be safely reinfused as early as 96 h after NOV administration."} +{"text": "In September 2010 the European Food Safety Authority (EFSA) released the findings of its latest review of bisphenol A (BPA), concluding there is no new evidence that warrants a revision of the current Tolerable Daily Intake of 0.05 mg/kg body weight.Significant new use rules went into effect 18 October 2010 for generic multi-walled carbon nanotubes and single-walled carbon nanotubes.3University of Southern California researchers have developed a smartphone application to estimate atmospheric particulate matter.EHP reported on the growing use of environmental stewardship claims in product marketing.7In June 2010, A new online resource offered by the U.S. EPA aids consumers battling bedbug infestations.9"} +{"text": "Arabidopsis imprinted genes FERTILIZATION INDEPENDENT SEED2 (FIS2) and FLOWERING WAGENINGEN (FWA) controlled by DNA methylation, and MEDEA (MEA) and PHERES1 (PHE1) controlled by histone methylation. Genome dosage imbalance deregulated the expression of FIS2 and PHE1 in an antagonistic manner. In addition increased dosage of inactive alleles caused a loss of imprinting of FIS2 and MEA. Although FIS2 controls histone methylation, which represses MEA and PHE1 expression, the changes of PHE1 and MEA expression could not be fully accounted for by the corresponding fluctuations of FIS2 expression. Our results show that parental genome dosage imbalance deregulates imprinting using mechanisms, which are independent from known regulators of imprinting. The complexity of the network of regulations between expressed and silenced alleles of imprinted genes activated in response to parental dosage imbalance does not support simple models derived from the parental conflict hypothesis.In mammals and in plants, parental genome dosage imbalance deregulates embryo growth and might be involved in reproductive isolation between emerging new species. Increased dosage of maternal genomes represses growth while an increased dosage of paternal genomes has the opposite effect. These observations led to the discovery of imprinted genes, which are expressed by a single parental allele. It was further proposed in the frame of the parental conflict theory that parental genome imbalances are directly mirrored by antagonistic regulations of imprinted genes encoding maternal growth inhibitors and paternal growth enhancers. However these hypotheses were never tested directly. Here, we investigated the effect of parental genome imbalance on the expression of Arabidopsis plants. Surprisingly, parental dosage imbalance affected each imprinted gene in a different manner and the imprinted status was also affected. Our results point to a relationship between imprinting and dosage imbalance that is more complex than predicted.In mammals and plants, imprinted genes are expressed preferentially by the copy inherited from either the mother or the father. In plants genome dosage is easily manipulated using tetraploid plants that contain twice the genome dosage of the natural diploid plants. The increased maternal dosage reduces seed size while increased paternal dosage has the opposite effect. It was further proposed that parental genomic imbalances are directly mirrored by antagonistic regulations of imprinted genes encoding maternal growth inhibitors and paternal growth enhancers. However these hypotheses were never tested directly. We measured the expression of imprinted genes and their regulators, in crosses between diploid and tetraploid MEA locus did not lead to unequivocal support Arabidopsis relatives Arabidopsis background remained to be tested in order to provide experimental evidence for the parental conflict theory in plants.In mammals and plants, mothers differentiate distinctive structures specialized in the transport of maternal nutrients to the embryo, the mammalian placenta and the plant seed endosperm Arabidopsis, the Polycomb Group (PcG) gene MEDEA (MEA) MATERNALLY EXPRESSED PAB C-TERMINAL (MPC) FERTILIZATION INDEPENDENT SEED 2 (FIS2) FWAFORMIN5MEA and FIS2 causes enhanced endosperm growth FORMIN5 loss of function leads to a reduction of endosperm growth and does not conform to the prediction of the parental conflict theory PHERES1 (PHE1) is a paternally expressed imprinted gene in Arabidopsis, which could play a role as a PEG ArabidopsisCurrently the regulation of five maternally expressed imprinted genes have been characterized in Arabidopsis, increased maternal genome dosage in seeds resulting from crosses between ovules from tetraploid plants and pollen from diploid plants (4nmat\u00d72npat) leads to production of smaller endosperm, embryo and seed mat\u00d74npat) cause the opposite effect. These results have suggested that collectively increased dosage of the expressed maternal allele of MIGs reduces endosperm growth while increased dosage of the expressed paternal allele of PEGs increases endosperm growth Plant reproduction is initiated by a double fertilization event MEA and PHE1 expression were regulated by FIS2MEA, FIS2, FWA and PHE1 in seeds resulting from interploid crosses. We performed quantitative RT-PCR to assess the expression of imprinted genes in endosperm produced by crosses between diploid and tetraploid plants and observed a global deregulation of expression levels of imprinted genes accompanied by an unexpected loss of parental imprinting for some genes. However the expression of known key regulators of imprinting were not affected. Our results suggest that parental dosage imbalance disrupts imprinting through interactions between imprinted genes and other unidentified regulators.Although it was assumed that parental dosage imbalances would be directly mirrored by variations in the expression of the PEGs and MEGs FIS2 and FWA. Conversely, the global level of expression of these genes should not be affected by an increased dosage of inactive paternal alleles. We used quantitative RT-PCR to investigate the effect of increased parental dosages in crosses between tetraploid and diploid plants. We measured the expression at 2 days after pollination (2DAP) when the imprinted genes studied are highly expressed and control the timing of endosperm development GAPC or more specifically in endosperm as MINI3Increased maternal dosage is expected to increase the level of expression of the active maternal allele of FIS2 and FWA, which are silenced by DNA methylation FWA expression crosses increased FIS2 mRNA levels in endosperm (mat\u00d74npat) crosses did not change the dosage of transcriptionaly active maternal FIS2 alleles, FIS2 expression was reduced in comparison to seeds produced by self-fertilized diploid plants. (FIS2 expression was reported from 3 to 5 DAP in Landsberg erecta background using the meiotic jason (jas) mutant, which produces a proportion of diploid pollen pFIS2-GUS, which contains the transcriptional regulatory cis-elements required for imprinting regulation FIS2 expression irrespective of its genomic context.We tested the effect of genome dosage imbalance on maternally expressed imprinted genes pression . By contmparable . As expendosperm . Surpris plants. . We obta plants. . A similArabidopsis and maize FIS2 expression and the down-regulation of FIS2 expression in response to increased dosage of inactive paternal alleles likely result from a distinct mechanism. We propose that the unexpected silencing of the maternal alleles of FIS2 in endosperm produced by (2nmat\u00d74npat) crosses could originate from increased paternal dosage of a paternally expressed imprinted inhibitor of FIS2 or from the activity of yet unidentified cis-elements.Trans-silencing in FIS2 and FWA in interploid crosses. Both genes remained strictly maternally expressed in (4nmat\u00d72npat) crosses (FIS2. The opposite (2nmat\u00d74npat) crosses did not affect the FWA imprinting status . We thus concluded that increased paternal dosage decreases the overall expression of FIS2 while both parental alleles become expressed. Such rather paradoxical effect is difficult to interpret. A negative interaction between MET1 activity, which maintains silencing marks and the trans-silencing mechanisms activated by the increased dosage of silenced paternal allele may cause removal of the silencing marks on the paternal allele of FIS2. Alternatively in response to reduced FIS2 expression, a transcriptional activator of FIS2 might be over-expressed and overcome silencing of the paternal allele.We assessed the imprinted status of crosses . This ing status but causprinting . Loss ofMEA and PHE1. MEA was predominantly expressed from the maternal allele in (4nmat\u00d72npat) crosses as in control diploid crosses (mat\u00d74npat) crosses the expression from the maternal allele decreased causing a predominant paternal expression of MEA leading to an apparent inversion of MEA imprinted expression crosses in C24 background crosses We further tested the effect of dosage imbalance on the maternally expressed imprinted gene ckground but not ckground . A modesckground . A mild PHE1 expression following the trends exhibited by MEA expression levels 12) Polycomb group subunit Su(z)12 family are not expressed in Arabidopsis endosperm FIS2 expression levels in (2nmat\u00d74npat) crosses could become limiting for Polycomb group activity, leading to increased expression of MEA and PHE1. This effect would also be directly responsible for the inversion of MEA imprinting (FERTILIZATION INDEPENDENT ENDOSPERM (FIE) FIE expression levels was kindly provided by Abed Chaudhury fis2-6 was previously identified in our laboratory The wild-type control lines C24, Col, Ler, and RLD were obtained from the ABRC stock center. The tetraploid lines in C24 and in Col ecotypes were kindly provided by Rod Scott Arabidopsis plants and frozen in liquid nitrogen. Total RNAs were extracted using the RNeasy mini kit (Qiagen). After DNAse treatment using DNase free kit (Ambion), RNAs were reverse-transcribed using Stratascript RT kit (Stratagene).Siliques two days after pollination (2DAP) were collected from http://rsbweb.nih.gov/ij/).Allele-specific RT-PCR reactions were performed as previously described ACT11 gene as endogenous control except for FIE was used as endogenous control. Relative Quantitation values (RQ) were calculated using the 2\u2212\u0394\u0394Ct method (RQ\u200a=\u200a2\u2212\u0394\u0394Ct) Real-time PCR assays were performed using a PCR Master Mix . One \u00b5l of RT product was used to perform each PCR reaction. Amplification reaction was carried out using specific primers at a concentration of 0.5 mM in a 10 \u00b5l reaction. Sequence of specific primer pairs can be found in Figure S1FIS2 (A), FWA (B), DME (C), MET1 (D), GAPC (E), and MINI3 (F) mRNAs were performed on total mRNAs extracted from siliques produced by crosses between diploid and tetraploid parents . Each point represents the average RQ value obtained for four independent biological samples (Effects of interploid crosses on the expression of DNA methylation dependent imprinted genes. (A-D) Quantitative PCR measurements of samples . Error b(0.89 MB TIF)Click here for additional data file.Figure S2FIS2 transgenes. Effect of an increased maternal dosage on expression of transcriptional reporter pFIS2-GUS expression at 1.5 DAP. Staining was stopped before signal saturation and three classes of seeds were distinguished on the basis of the intensity of signal. The percentage of each class in crosses between ovules of the marker line and wild-type pollen from diploid or tetraploid plants is indicated below each corresponding micrograph. Scale bars correspond to 25 \u00b5m.Effects of interploid crosses on the expression (2.58 MB TIF)Click here for additional data file.Figure S3MEA (A), PHE1 (B), and FIE (C) mRNAs were performed on total mRNAs extracted from siliques produced by crosses between diploid and tetraploid parents . Each point represents the average RQ value obtained for four independent biological samples (Effects of interploid crosses on the expression of genes imprinted by Polycomb group activity. Quantitative PCR measurements of samples . Error b(0.75 MB TIF)Click here for additional data file.Table S1Act11. C24 2nX2n sample 1 was normalised to 1. Each RQ value in this table corresponds to the average RQ value of 3 technical replicates.RQ value of the C24 experiment after normalisation with (1.32 MB TIF)Click here for additional data file.Table S2Probability values obtained after a student's t-test on C24 sets of crosses from (1.31 MB TIF)Click here for additional data file.Table S3Act11. Col 2nX2n sample 1 was normalised to 1. Each RQ value in this table corresponds to the average RQ value of 3 technical replicates.RQ value of the Columbia experiment after normalisation with (1.34 MB TIF)Click here for additional data file.Table S4Probability values obtained after a student's t-test on Col sets of crosses from (1.31 MB TIF)Click here for additional data file.Table S5Probability values obtained after a student's t-test on qPCR results shown in (0.76 MB TIF)Click here for additional data file.Table S6List of primers used in this study.(1.12 MB TIF)Click here for additional data file."} +{"text": "P = 0.64). Disease progression occurred within a year in about 50% of patients who were initially untreated. Response rate was similar in both groups, but duration of response was shorter in patients who were treated at disease progression . Patients actually treated at disease progression (34/70) survived shorter than those who had neither disease progression nor treatment . Starting MPH-P just after diagnosis does not improve survival and response rate in stage I MM, with respect to deferring therapy until disease progression. However, patients with stage I MM randomized to have treatment delayed and who actually progressed and were treated had shorter survival than those with stable disease and no treatment. Biologic or other disease features could identify these subgroups of patients. \u00a9 2000 Cancer Research CampaignWe conducted a randomized trial to evaluate whether melphalan-prednisone (MPH-P) treatment administered just after diagnosis improves survival of stage I multiple myeloma (MM). Between January 1987 and March 1993, 145 consecutive previously untreated patients with stage I MM were randomized between treatment with MPH-P (administered for 4 days every 6 weeks) just after diagnosis and treatment only at disease progression. Survival was not influenced by MPH-P treatment either administered just after diagnosis or at disease progression (64 vs 71 months respectively). Comparing the first with the second group the odds ratio of death is 1.17 (95% confidence interval 0.57\u20132.42;"} +{"text": "In previous studies, highly heterogeneous uptake of 131I-labelled chimeric monoclonal antibody G250 ([131I]cG250) in primary renal cell carcinomas has been observed . In this study, we investigated a possible correlation between intratumoral antibody uptake and four immunohistochemically determined parameters: G250 antigen expression, blood vessel density, neovascularization and percentage of viable tumour cells. Whole tumour slices of four different tumours were cut into 1-cm3 cubes, and in each cube the [131I]cG250 uptake was determined. The correlation between [131I]cG250 uptake and each individual parameter was determined in a multiple regression analysis. Additionally, the data were reanalysed after introducing arbitrary cut-off values for each parameter. If a sample showed expression of a parameter above the introduced threshold value, this sample fulfilled one condition. Subsequently, the Pearson correlation coefficients were calculated from [131I]cG250 uptake and the number of fulfilled conditions (0-3). All tumour samples with high [131I]cG250 uptake [> 0.1% of the injected dose per gram (ID g(-1))] showed high antigen expression (> 50%). However, not all samples with high antigen expression displayed high uptake. A statistically significant correlation between [131I]cG250 uptake and antigen expression was found in three out of four tumours analysed. Of the other determined parameters, no consistent correlation with [131I]cG250 uptake was found; only the percentage of viable tumour cells correlated significantly in two out of four tumours (beta = 0.80 and 0.26). Calculation of the Pearson correlation coefficients showed a statistically significant correlation between [131I]cG250 uptake and an increased number of fulfilled conditions in all tumours, indicating that each of the individual parameters contribute to the uptake of [131I]cG250. These observations indicate that high antigen expression is a prerequisite for high antibody uptake. However, regional differences in antibody uptake within a tumour cannot be explained by antigen expression alone."} +{"text": "The relationship between vascular endothelial growth factor (VEGF) and lymph node metastasis was studied in 90 cases of primary lung cancer without distant metastasis. As a result of quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis, the VEGF121 mRNA expression levels in lung cancer tissues with nodal metastasis (n = 35) were higher than in those without nodal metastasis (n = 55). However, no significant difference could be found in VEGF121 mRNA expression levels as stratified by tumour size (T1N0M0 vs T2N0M0). Simultaneously, ten lymph nodes (four node positive and six node negative) together with the corresponding primary lung tumours and adjacent normal lung tissue, were studied for VEGF expression. The VEGF mRNA expression in metastatic lymph nodes was intense in three out of the four cases examined. Further, while VEGF expression levels in metastatic lymph nodes were conspicuously higher than those for the primary site, all its expression levels in non-metastatic nodes were inferior to those of the primary tumours. Except for macrophages, the VEGF antigen was identified mainly in the cytoplasm of metastatic cancer cells and the endothelial cells of blood or lymphatic vessels in lymph nodes. Although the detailed mechanisms and the significance of strong VEGF expressions in metastatic lymph nodes are still unknown, these data are consistent with a model whereby VEGF increases the opportunity for nodal metastasis through neoblood and lymphatic vessels."} +{"text": "Current reports suggest serum S100 as a prognostic marker for disease progression in advanced malignant melanoma. In this study, we assessed serum levels of S100 and multiple clinical factors in relation to overall survival in 99 patients with metastatic malignant melanoma seen at our institution between May 1990 and April 1996. For statistical analysis, we used both univariate and multivariate Cox proportional-hazards models. Elevated serum levels of S100 correlated with poor outcome in metastatic malignant melanoma (P < 0.0001), univariate analysis). Upon multivariate analysis, however, S100 added no information to known clinical prognostic parameters."} +{"text": "Patients undergoing surgical treatment for calculous disease were considered to have had a partialcholecystectomy performed when a part of the gall bladder wall was retained for technical reasons.Forty patients underwent partial cholecystectomy: for chronic cholecystitis (20), acute cholecystitis (4),Mirizzi's syndrome (14), portal hypertension or partially accesible gall bladder (one patient each). Fourpatients (10%) developed infective complications and two patients had retained common bile ductstones. In a mean follow up period of 13 months (range 1\u201336 mths), only 3 patients have ongoing milddyspeptic symptoms while the rest have remained asymptomatic. Partial cholecystectomy has beenfound to be a safe and effective procedure in difficult cholecystectomy situations, since it combines themerits of cholecystectomy and cholecystostomy."} +{"text": "Seventy-eight adult patients with acute leukaemia were classified cytologically into 3 categories: acute lymphoblastic leukaemia (ALL), acute myelogenous leukaemia (AML) or acute undifferentiated leukaemia (AUL). The periodic acid-Schiff stain was of little value in differentiating the 3 groups. The treatment response in each group was different: 94% of patients with ALL (16/17) achieved complete remission with prednisone, vincristine and other drugs in standard use in childhood ALL; 59% of patients with AML (27/46) achieved complete remission with cytosine arabinoside and daunorubicin (22 patients), or 6-thioguanine and cyclophosphamide (2 patients), 6-thioguanine, cyclophosphamide and Adriamycin (1 patient), and cytosine and Adriamycin (1 patient); only 2 out of 14 patients (14%) with acute undifferentiated leukaemia achieved complete remission using cytosine and daunorubicin after an initial trial of prednisone and vincristine had failed. Prednisone and vincristine would seem to be of no value in acute undifferentiated leukaemia. It would seem also that no benefit is obtained by classifying all patients with acute leukaemia over 20 years of age as \u201cadult acute leukaemia\u201d and treating them with the same polypharmaceutical regimen. The problems posed by each disease are different and such a policy serves only to obscure them."} +{"text": "Response to preoperative radiochemotherapy (RCT) in patients with locally advanced rectal cancer is very heterogeneous. Pathologic complete response (pCR) is accompanied by a favorable outcome. However, most patients show incomplete response. The aim of this investigation was to find indications for risk stratification in the group of intermediate responders to RCT.From a prospective database of 496 patients with rectal adenocarcinoma, 107 patients with stage II/III cancers and intermediate response to preoperative 5-FU based RCT (ypT2/3 and TRG 2/3), treated within the German Rectal Cancer Trials were studied. Surgical treatment comprised curative (R0) total mesorectal excision (TME) in all cases. In 95 patients available for statistical analyses, residual transmural infiltration of the mesorectal compartment, nodal involvement and histolologic tumor grading were investigated for their prognostic impact on disease-free (DFS) and overall survival (OS).Residual tumor transgression into the mesorectal compartment (ypT3) did not influence DFS and OS rates . Nodal involvement after preoperative RCT (ypN1/2) turned out to be a valid prognostic factor with decreased DFS and OS . Persistent tumor infiltration of the mesorectum (ypT3) and histologic tumor grading of residual tumor cell clusters were strongly correlated with lymph node metastases after neoadjuvant treatment (p < 0.001).Advanced transmural tumor invasion after RCT does not affect prognosis when curative (R0) resection is achievable. Residual nodal status is the most important predictor of individual outcome in intermediate responders to preoperative RCT. Furthermore, ypT stage and tumor grading turn out to be additional auxiliary factors. Future clinical trials for risk-adapted adjuvant therapy should be based on a synopsis of clinicopathologic parameters. Multimodal treatment strategies and optimized surgical procedures with total mesorectal excision (TME) led to a significant improvement in rectal cancer therapy within the last 15 years ,7.After preoperative RCT, therapy-induced downsizing effects have widely been described as important prognostic factors ,9. Local(y)pT3a to (y)pT3d. Therefore (y)pT3 category spans the invasion of only a few tumor cells beyond the muscularis propria to a complete infiltration of the mesorectum, nearby reaching the visceral peritoneum or contiguous organs [According to TNM classification , tumor is organs .A risk-adapted stratification of patients after preoperative RCT and TME-based surgery is crucial for adjuvant treatment strategies in individual patients. Currently, a beneficial impact of adjuvant chemotherapy (CT) is discussed controversely ,16. To dIn this study we investigated 107 patients with intermediate local response to preoperative 5-FU based RCT (ypT2/3) and curative (R0) surgery. The aim of this investigation was to clarify the impact of residual tumor infiltration of the mesorectal compartment (\u2265 ypT3b), nodal status (ypN) and histologic tumor grading on DFS and OS and to evaluate their relevance within an individual risk stratification model in intermediate responders to RCT.This study included patients with locally advanced rectal cancer (stage II/III) and moderate RCT-induced histopathologic tumor regression (TRG 2 and 3 according to the Dworak classification) and conPatients with clinical evidence of distant metastatic disease were excluded from the actual investigation and received individual multimodal treatment.Pretherapeutical staging procedures consisted of rigid rectoscopy, flexible colonoscopy, endorectal ultrasound (ErUS), magnetic resonance imaging (MRI) of the pelvis and computed tomography (CT) scans of chest, liver and pelvis. Staging results were conferred and interdisciplinary discussed before initiation of multimodal treatment. Clinical tumor stages were determined by ErUS, pelvic MRI, and CT scans.Preoperative treatment included fractional radiation with cumulative 50.4 Gy (28 \u00d7 1.8 Gy) in 3- or 4-field technique. Concomitant chemotherapy consisted of either 5-Fluorouracil (5-FU) monotherapy in 84 patients or a combined 5-FU + Oxaliplatin regime in 23 patients. Six weeks after completion of neoadjuvant treatment all patients underwent standardized TME-based surgery. Subsequently, postoperative systemic therapy was applied according to the preoperative treatment regimen and the actual study protocol.Quality assessment of the surgical specimens was performed according to the MERCURY criteria and was Pathological staging included ypTNM stage according to the current TME classification, tumor dresidual tumor cells was evaluated after preoperative RCT and subdivided into two categories by a Cox proportional hazards regression model. The ypT and ypN parameters were additionally evaluated in a multivariate analysis.http://www.r-project.org).The distributions of ypN status within the two subgroups of ypT (ypT2/3a and ypT3b-d) were compared by Fisher's exact test. The number of detected lymph nodes between nodal positive and negative patients was compared with the Mann-Whitney-U test. The significance level was set to \u03b1 = 5% for all tests. All analyses were performed with the free software R rectal cancer received preoperative RCT within the German rectal cancer trials , as detected by manual liver palpation and intraoperative ultrasound. These patients likewise had evidence of residual mesorectal lymph node metastases within the surgical specimen and were excluded from survival analysis.During a median follow-up period of 42 months (range: 4 - 126 month), 9 of the 107 patients died of non-cancer-related disease and were excluded from cancer specific survival analyses. Seventeen (15.9%) patients had cancer relapse, with 15 cases of separate distant metastatic disease and 2 cases of local recurrence combined with synchronous distant metastases. No isolated local recurrence occurred. Three patients failed statistical analyses due to occult synchronous hepatic metastases detected during surgery (ypUICC IV). In summary, 95 patients were included into survival analysis.Patient characteristics, pretherapeuthical staging results and treatment procedures of all 107 intermediate responders to preoperative RCT are presented in Table When comparing pretherapeuthical clinical staging with pathological staging results, RCT-induced T-Level downsizing was achieved in 42% of patients (n = 45). Eight tumors, initially staged as cT4 were downsized to ypT2 (n = 2), ypT3b (n = 2), ypT3c (n = 3) and ypT3d (n = 1). Thirty-seven tumors, previously staged as cT3 were downsized to ypT2 status. Nodal downstaging from cUICC III to ypUICC II stage was achieved in 41% of patients (n = 44).The median number of detected and histopathologically evaluated lymph nodes per specimen was 21 (range: 6 - 79). In 68% of specimens, lymph node yield accounted for \u2265 18 nodes. Fewer than 12 nodes, which is the consensual number according to TNM criteria, were found in a total of 5 specimens (4.7%).In patients with extended lymph node recovery, lymph node metastases were detected more frequently. However, this finding was not statistically significant (p = 0.06). In detail, the median number of lymph nodes found in the ypN0 group was 19 (range: 6 - 79) compared to 24 (range: 7 - 77) in the ypN1/2 group.Of 95 patients included in cancer specific survival analyses, 63 (66.3%) were classified as node negative (ypN0), and 32 (33.7%) patients presented with residual lymph node metastases (ypN1/2) after RCT. Fifty patients (54.3%) had intramural tumor infiltration with a maximal infiltration of \u2264 1 mm beyond the muscularis propria (ypT2/3a). Forty-two patients (45.7%) had advanced ypT status with distinct (>1 mm) tumor invasion into the mesorectal compartment (ypT3b-d).Compared to the ypT2/3a stage, advanced residual infiltration into the mesorectal compartment ypT3b-d) after preoperative RCT was not associated with a significantly decreased DFS and OS Figure . However-d after The probability of cancer relapse and distant metastases was stage-dependent. There was no significant difference within the group of nodal-negative patients with stage I and II disease (ypT2/3a N0 and ypT3b-d N0: 91% and 88%) or within the group of stage III patients (ypT2/3a N+ and ypT3a-d N+: 55% and 67%).Residual mesorectal tumor infiltration (ypT3b-d) - though without immediate impact on survival - was significantly associated with occurrence of metastatic lymph node involvement after preoperative RCT (p < 0.001), which itself is an independent prognostic factor for survival.Histologic tumor differentiation grading after RCT had a significant influence on DFS (p = 0.04), whereas patients with well and moderate tumor differentiation showed a tendency for prolonged OS . Furthermore, histologic tumor differentiation grading after RCT correlated with residual lymph node metastases (p < 0.001) as well as mesorectal tumor infiltration (ypT3b-d) (p = 0.0001).When evaluating the 63 patients with ypN0 status for their pretherapeuthical nodal status (cN), staged by ErUS and MRI, 29% (n = 18) of patients had previous cN0 status and 71% (n = 45) presented with cN+ status. DFS and OS did not significantly differ in patients who initially presented with clinical evidence of mesorectal lymph node involvement but resulted in ypN0 after RCT . Patients with clinically staged III rectal cancers therefore showed no higher risk of cancer relapse and cancer-related death than initially node-negative patients, as long as sterilization of lymph node metastases can be achieved with RCT.Recent results from the randomized multicenter trial CAO/ARO/AIO-94 showed an enhanced local control and sphincter preservation with concurrently decreased toxicity after preoperative long-term RCT compared to postoperative RCT . These rOf 153 patients with stage II/III rectal cancer who received standardized preoperative RCT within randomized clinical trials, pCR as a major response criterion, was achieved in 16% n = 10) of patients. pCR rates vary between 10 and 20% and were associated with a favorable outcome ,10. Neve of patieIt remains unclear which subgroup of patients with intermediate response can be considered as cured after preoperative RCT and subsequent TME surgery. Conversly, it is of enormous clinical interest to know which subgroup necessitates adjuvant systemic therapy.Involvement of circumferential resection margins (CRM) has recently been described as a very strong prognostic factor after preoperative short term radiation . AlthougPrior to implementation of neoadjuvant strategies for rectal cancer, a tumor invasion of \u2265 5 mm into the mesorectal compartment, besides circumferential involvement, was described as a significant prognostic factor . The decWe therefore evaluated the impact of intramural depth of tumor invasion (ypT2) together with minimal (<1 mm) transgression of the muscularis propria (ypT3a) compared to a distinct transmural tumor invasion into the mesorectum . Since patients with ypT3a status show only an extremely marginal infiltration of the mesorectal compartment (<1 mm) we consider them to prognostically belong to the ypT2 group rather than to the tumors with distict mesorectal infiltration. Our results underline this assumption showing an increased incidence of nodal metastases in ypT3b-d patients compared to ypT2/3a patients.In the patients presenting with previous cT3/4 rectal cancers the RCT-induced regression of tumor invasion depth to ypT2/3a status had no impact on prolonged DFS and OS. Thus, residual tumor transgression into the mesorectum after preoperative RCT showed no significant influence on cancer recurrence, providing that complete resection with negative CRM is achieved by adequate TME surgery.Tumor downsizing from the extramural mesorectal compartment into the actual rectal wall therefore seems to be of importance only when tumor-free CRM and R0-resection cannot be guaranteed (former T3d/4 status).In contrast to ypT, nodal status after preoperative CRT (ypN) significantly influenced cancer recurrence and overall survival in stage II/III rectal cancer patients with intermediate response within our investigation. This finding coincides with previous results and supports recent investigations with considerable numbers of patients ,26 but iIn agreement with other authors ,27, we oAnyway, patients with ypN+ status should be considered for upcoming trials with intensified adjuvant CT regimes as this might be more efficient in preventing systemic tumor relapse. Nonetheless, mesorectal tumor invasion (ypT3b-d) was significantly associated with residual lymph node metastases after RCT in our study (p < 0.001). We interpret this finding with a generally lower response to RCT regarding both downsizing of the primary tumor and sterilization of lymph node metastases. This might be due to improved biological behavior and enhanced resistance to RCT in individual cancers. The prognostic impact of mesorectal tumor infiltration remains unclear. We could not show straight effects on tumor recurrence and survival but are well aware that this might be due to the relative small number of patients underlying this investigation.Neoadjuvant RCT has repeatedly been accused of reducing lymph node yield in rectal cancer specimens -31. It hWhile histologic tumor grading in colorectal cancers after primary surgery has been ascertained as a prognostic factor , its proNot unexpectedly, lymph node status displays as the major criterion for therapy stratification after application of preoperative RCT within our study and several recent investigations and might subdivide patients with need of intensified adjuvant treatment from those who can be considered as cured after surgery. In contrast Collette et al. , who repTo date, most patients with positive nodal status after preoperative RCT will intuitively get adjuvant CT. Prospective randomized clinical trials should therefore clarify the impact of adjuvant treatment in patients undergoing preoperative RCT and radical surgery. For ypN0 patients 5-FU based adjuvant CT was shown as a potential overtreatment and had no significant effect on survival ,37.Nevertheless, in our actual study population, 7 patients with ypN0 status developed distant metastases during follow-up. All 7 had poorly differentiated residual tumors (low grade). Poor differentiation of residual tumor cell clusters after RCT and advanced invasion depth turned out to be predictors of lymph node metastases and may be indicators of occult nodal (micro-) metastases in patients classified as ypN0. Both parameters should thus be taken into account in ypN0 patients, particularly in cases of minor lymph node recovery, and might have influence on the decision for adjuvant CT.Although this investigation is based on a homogeneous collective of patients treated within randomized clinical trials with replicable and standardized diagnostic and therapeutic procedures, its principal limitations are the retrospective character and the relatively small number of patients. Thus this study does not want to claim to ultimately answer the question which subgroup of patients need adjuvant CT after preoperative multimodal treatment and subsequent R0-resection. Prospective randomized trials will have to clarify the debatable role of postoperative CT in rectal cancer patients after preoperative RCT and radical TME surgery. The clinicopathologic parameters investigated in this study might give indications to stratify patient groups with lower and higher individual risk of tumor relapse and tumor-related death within future clinical trials.The authors declare that they have no competing interests.TS prepared the study design, assembled and analysed the data and drafted the manuscript. HR carried out the pathological diagnostics of the rectal cancer specimens and reviewed the manuscript. KJ carried out the statistical analyses. HC contributed the radiation therapy data and reviewed the manuscript. LC participated in assembling of the data and reviewed the manuscript. BMG and HB reviewed the manuscript. TL supervised the study and data assembling and critically reviewed the manuscript. All authors read and approved the final manuscript."} +{"text": "Acute cholangitis is associated with a high mortality and morbidity and often requiresdrainage of the obstructed biliary system. The purpose of this study was to evaluate theusefulness and safety of endoscopic nasobiliary drainage in the treatment and preventionof acute cholangitis due to diverse etiology. During a 32-month period, 143 patients with age range of 15 to 84 years underwent urgent fluoroscopyguided endoscopic nasobiliary drainage using a 7 Fr catheter either to treat acutecholangitis not responding to antibiotics or to prevent its developmentfollowing endoscopic retrograde cholangiography performed in an obstructed biliarysystem . Underlying etiology included bile duct stones (92), malignantbiliary obstruction (34), choledochal cyst (4), chronic pancreatitis (4), ruptured hydatidcyst (3), portal hypertensive cholangiopathy (3) and liver abscess (3). Endoscopicnasobiliary drainage was performed successfully in 129 patients (90.2%). Cholangitisimproved within 1 to 3 days (in group A) or did not develop (in Group B) in 125 patients(96.7%) with successful endoscopic nasobiliary drainage. Two patients however requiredadditional drainage by percutaneous transhepatic route, while two died inspite of effectiveendoscopic drainage. Of the 14 patients (9.8%) with failed endoscopic drainage, 9 weremanaged by surgical decompression or percutaneous transhepatic drainage, 3 died ofsepticemia. Endoscopic nasobiliary drainage is a safe and effective method to treatpatients with acute cholangitis as well as to prevent its development followingcholangiography performed in an obstructed biliary system."} +{"text": "In In There are errors in the There are errors in the There are errors in the There are errors in the There are errors in the There are errors in the There are errors in S1 TableMonthly under 5 mortality ratio ((U5MR); Deaths before 60 months of age per 1000 live births) or gender mortality (count of monthly deaths before 153 days) regressed on MEAN monthly temp and MEAN temp in the prior month. All models use first differences of all variables to correct for non- stationarity. ARIMA terms included to minimize AIC.(DOCX)Click here for additional data file.S2 TableMonthly under 5 mortality ratio (Deaths before 60 months of age per 1000 live births) or gender mortality (count of monthly deaths before 153 days) regressed on MAXIMUM monthly temp and MAXIMUM temp in the prior month. All models use first differences of all variables to correct for non-stationarity.(DOCX)Click here for additional data file.S3 TableMonthly neonatal mortality (Death count before 1 month) and monthly post neonatal mortality (Death count between 30 and 153 days) regressed on MEAN monthly temp and MEAN temp in the prior month. All models use first differences of all variables to correct for non- stationarity. ARIMA terms included to minimize AIC. Both sexes analysed together.(DOCX)Click here for additional data file.S4 TableMonthly neonatal mortality (Deaths before 1 month) and monthly post neonatal mortality (Death count between 30 and 153 days) regressed on MEAN monthly temp and MEAN temp in the prior month. All models use first differences of all variables to correct for non- stationarity. ARIMA terms included to minimize AIC. Both sexes analysed together.(DOCX)Click here for additional data file.S5 TableTable S5 below shows the AIC of several ARMA models fitted to the residuals (e1) of the various regressions at time lag = 0(DOCX)Click here for additional data file.S6 TableTable S6 below shows AIC of ARMA models fitted to the residuals (e2) of the various regressions at time lag = 1(DOCX)Click here for additional data file.S1 FileMonthly reports of temperature, maximum temperature, minumum temperature, infant mortality, female infant mortality, male infant mortality, male mortality less than 30 days, male mortality greater than 30 days, female mortality less than 30 days, female mortality greater than 30 days, both sex mortality less than 30 days, and both sex mortality greater than 30 days.(XLS)Click here for additional data file."} +{"text": "The experimental procedure for the grafting of POEGMA and PAAm via atom transfer radical polymerization (ATRP) is described in Wassel et al. (2019) https://doi.org/10.1016/j.matdes.2018.107542 [1]. The FTIR spectra of the porous oxides before and after attachment of (3-Aminopropyl)trimethoxysilane (APTMS) are presented. Microscopic images of thick POEGMA films and PAAm on AAO are displayed, and an FTIR spectrum of AAO/PAAm is shown. An EDX mapping of carbon is shown on an AAO/POEGMA sample. The adsorption behavior of Fluorescein isothiocyanate (FITC) marked bovine serum albumin (BSA) on patterned porous TiO2-NT films is documented. Finally microscopic images are presented to compare the scratch resistance behavior of pristine porous films with those functionalized with POEGMA.The data presented in this article affords insight into the fabrication and ensuing microstructure of the supported porous anodic aluminum oxide (AAO) and TiO Specifications tableValue of the data\u25cfThese data demonstrate the successful grafting of antifouling poly(oligo ethyleneglycol) methylether methacrylate (POEGMA) and acrylamide (AAm) brushes directly onto the pore walls of supported porous anodized aluminum oxide (AAO), generating robust transparent organic-inorganic anti-adhesive nanocomposite films that can be used for displays and as windows for submerse optical sensors.\u25cf2-nanotubes (NT), yielding 3D anti-adhesive surfaces that can be used for Ti-base surgical implants. These porous oxide materials afford a huge surface area.Our data also verify the covalent-grafting of antifouling polymer brushes onto the pore walls of TiO\u25cf2-NT films were investigated and proved using Fluorescein isothiocyanate (FITC) marked bovine serum albumin (BSA). The data obtained point to the strong potential of our 3D anti-fouling coatings as candidates in the area of health and biomedical research.The anti-adhesive properties of anchored POEGMA brushes into the patterned 3D porous TiO1Data presented in this article displays a drawing showing the test procedure for the scratch test .Fig. 1A 2-NT.The SEM images show top views and cross sections of pristine AAO pores superimposed on a secondary electron micrograph is displayed in 2 surface ) demonstrate the similar grafting of PAAm polymer brushes on the surface and inside of the pores.Not only the obtained SEM micrograph proves tSecondary electron micrographs and EDX analyses of AAO/POEGMA before 2 surfaces before and after grafting of OEGMA brushes (Different wettability of the AAO and TiO2-structured surface and subsequently treated with FITC labelled BSA (The Fluorescence micrograph obtained from a TiOlled BSA shows gr2-NT (4\u2009N) with APTMS known as linking moieties for attachment of the atom transfer radical polymerization (ATRP)-initiator for the synthesis of polymer brushes.The modification of AAO and TiO2.4POEGMA brushes were prepared using BiPy (1.2480\u2009g), 12\u2009mL of deionized water and methanol (1:4), purified OEGMA (19.08\u2009mL) and CuBr (0.5720\u2009g), while for synthesis of PAAm brushes PMDETA, (0.28\u2009mL), 20\u2009mL of deionized water and methanol (3:7), AAm (2.00\u2009g) and CuBr (0.064\u2009g) were used.2.5Structured samples were incubated in aqueous solution of FITC-labelled BSA (3\u2009mg/mL) for 2\u2009hours at 37\u2009\u00b0C, rinsed with water, dried with nitrogen gas and investigated using a fluorescence microscope.2.6For the scratch test a hardened steel sphere was moved on the surface with a constant load, describing a meander as in 2.7Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), Water contact angle (WCA) and Scanning electron microscopy were used to characterize material chemistry and microstructure, wettability and topography of the pristine - and covered surfaces with polymer brushes."} +{"text": "Seeds of plants are a confirmation of their next generation and come associated with a unique microbia community. Vertical transmission of this microbiota signifies the importance of these organisms for a healthy seedling and thus a healthier next generation for both symbionts. Seed endophytic bacterial community composition is guided by plant genotype and many environmental factors. In north-east India, within a narrow geographical region, several indigenous rice genotypes are cultivated across broad agroecosystems having standing water in fields ranging from 0-2 m during their peak growth stage. Here we tried to trap the effect of rice genotypes and agroecosystems where they are cultivated on the rice seed microbiota. We used culturable and metagenomics approaches to explore the seed endophytic bacterial diversity of seven rice genotypes (8 replicate hills) grown across three agroecosystems.P\u22640.05) across genotypes. Significant differences were also found between the individual hills of a rice genotype. PCoA analysis exhibited three separate clusters and revealed the clusters separated based on genotype, while agroecosystem showed a minimal effect on the variation of seed microbiota . Interestingly, animal gut resident bacteria such as Bifidobacterium, Faecalibacterium, Lactobacillus, etc. were found in abundance as members of the seed microbiota.From seven growth media, 16 different species of culturable EB were isolated. A predictive metabolic pathway analysis of the EB showed the presence of many plant growth promoting traits such as siroheme synthesis, nitrate reduction, phosphate acquisition, etc. Vitamin B12 biosynthesis restricted to bacteria and archaea; pathways were also detected in the EB of two landraces. Analysis of 522,134 filtered metagenomic sequencing reads obtained from seed samples (n=56) gave 4061 OTUs. Alpha diversity indices showed significant differences in observed OTU richness (Overall, our study demonstrates, indigenous rice genotypes of north-east India have a unique blend of endophytic bacteria in their mature seeds. While there are notable variations among plants of the same genotype, we found similarities among genotypes cultivated in completely different environmental conditions. The beta diversity variations across the seven rice genotypes were significantly shaped by their genotype rather than their agroecosystems. The occurrence of bacteria in the rhizosphere and internal parts of plants is a natural phenomenon \u20134. Sever(Oryza sativa), and this region is home to many local landraces of rice. The cultivated rice has originated from its wild ancestors 10,000 years ago based on evidence gathered from studies using phylogeographic approach . Agar plates were incubated at 28\u2218C and observed for bacterial growth till seven days. The number of colony forming units (cfu) from each replicate was counted under a colony counter and reported in log cfu/g dry seed (DS). Each replicate value of a dilution was multiplied with the dilution factor and then calculated for 1 gram sample. After conversion to log10 scale, mean was calculated, standard error was determined and bar graph was plotted in MS Excel , 3 \u03bcl 10% SDS and 3 \u03bcl proteinase K (20 mg/ml) were added and incubated for 1 hour/ 60\u2218C. 100 \u03bcl 5 M NaCl was added followed by 80 \u03bcl of CTAB/NaCl solution (10% CTAB in 0.7 M NaCl), mixed and incubated for 10 min/ 65\u2218C. Phenol: chloroform: isoamyl alcohol (25:24:1) was added for extraction followed by one volume of chloroform: isoamyl alcohol (24:1) to the aqueous phase. DNA was precipitated with 5 M NaCl and two volumes of chilled absolute ethanol. The pellet was washed twice with 80% ethanol. The dried pellet was resuspended in TE buffer (pH 8.0). RNase treatment was done as required. Samples were checked in agarose gel. 16S rRNA gene was amplified in Eppendorf Mastercycler using primer pair 27f (5\u2019 - AGAGTTTGATYMTGGCTCAG) and 1492r (5\u2019 - TACCTTGTTAYGACTT). PCR products were purified using Gen-elute PCR cleanup kit (Sigma). Purified samples were sequenced in Applied Biosystems sequencer at Xcelris Genomics, Ahmedabad, Gujarat. Two additional internal primers, 533f (5\u2019- GTGCCAGCAGCCGCGGTAA), 805r (5\u2019- GACTACCAGGGTATCTAATCC) were used for sequencing. Contigs were assembled based on their phred scores (>15) and identified by aligning in NCBI reference rRNA database using the blastn algorithm. 16S rRNA gene sequences were used to build a phylogenetic tree in MEGA7 software [Prevotella brevis strain GA33 was used as outgroup. Sequences from isolates of Maguri bao had very low phred scores and could not be identified.The method by Ding et al. was follsoftware to deterThe 16S rRNA gene sequences were individually aligned in PAPRICA (Pathway Prediction by Phylogenetic Placement) pipelineMetagenomic DNA was extracted from 1 gram dehusked and surface sterilized seeds of the 8 hills of seven genotypes. Each hill was treated as a biological replicate. DNA was extracted by the method of Sharma et al. For nextThe demultiplexed data was processed in QIIME v1.9.1 . Paired-Additional file 1Table S1 number of colony forming units in each replicate of the seven culture mediaacross the seven genotypes and statistical analysis.Additional file 2Table S2 list of metabolic pathways and their confidence score across the culturable solates.Additional file 3Table S3 next generation sequencing based OTUs of endophytic bacteria at different taxa and percentage abundance of unaasigned OTUs.Additional file 4Table S4 range of three alpha diversity indices in the seven rice genotypes.Additional file 5Table S5: list of OTUs detected in all the hiils of a genotype."} +{"text": "This study evaluated the clinical, laboratory, ultrasonographic and pathological findings in 87 cows aged 2 to 10\u2009years (4.5\u2009\u00b1\u20091.5\u2009years) with type-4 abomasal ulcer.The most common clinical findings were in decreasing order compromised health status accompanied by partial or complete anorexia (100%), abdominal guarding (81%), congested scleral vessels (77%), ruminal atony (73%), tachycardia (68%), tachypnoea (65%), positive foreign body tests (58%), decreased skin surface temperature (53%), fever (49%), reduction in negative intraabdominal pressure assessed transrectally (39%), poorly subdivided plant fragments in faeces (35%) and arched back (28%). The principal haematological abnormalities were hypokalaemia (72%), haemoconcentration (69%), azotaemia (56%), metabolic acidosis (49%), hyperfibrinogenaemia (45%), leukopenia (35%) and hypoproteinaemia (29%). Other abnormalities were aciduria (56%), haematuria (44%), increased chloride concentration in rumen fluid (34%) and abnormal peritoneal fluid (98%). Of 75 examined cows, 65 (87%) had ultrasonographic evidence of local or generalised peritonitis. On postmortem examination all cows had a type-4 abomasal ulcer and generalised peritonitis. In addition, 36 cows had type-1 ulcers, 6 had type-2 ulcers and one cow had a type-3 ulcer.The clinical signs in cows with type-4 abomasal ulcer are associated with generalised peritonitis. An increased haematocrit, indicating shock-induced haemoconcentration is characteristic in contrast to cows with traumatic reticuloperitonitis. Ultrasonography is useful for visualising and assessing generalised peritonitis.The diagnosis of type-4 abomasal ulcer based on clinical signs alone is difficult and therefore requires additional diagnostic procedures including the determination of the haematocrit and plasma protein concentration, abdominal ultrasonography and analysis of peritoneal fluid. In most cases, these steps lead to a correct diagnosis and allow timely euthanasia of the cow to prevent further suffering and unnecessary treatment costs.The cows underwent a clinical, laboratory, ultrasonographic and postmortem examination. Abomasal ulcer disease is of great importance in cattle. Abomasal lesions are divided into erosions and ulcers ; erosionThe clinical signs vary widely depending on the type of abomasal ulcer; generalised peritonitis is common in cows with type-4 ulcer and is often fatal within 24 to 48\u2009h . The cliUltrasound examination can be helpful in assessing the position, size, wall and content of the abomasum and possible inflammatory lesions that involve neighbouring organs . HoweverBecause the clinical diagnosis of type-4 abomasal ulcer is not straightforward, and detailed investigations of large numbers of cows with this ulcer type are lacking, the purpose of this study was to describe the clinical, laboratory and ultrasonographic findings of cows with type-4 abomasal ulcer to facilitate the diagnosis of this disease.n\u2009=\u200936, 49%) became ill within 4\u2009weeks after calving . Twenty-four cows had been treated with non-steroidal anti-inflammatory drugs (NSAIDs), six with corticosteroids and another six with NSAIDs and corticosteroids before referral, but the exact dosages were not known.This was a retrospective study of 87 cows that had a main diagnosis of type-4 abomasal ulcer. The cows had been admitted to the Veterinary Teaching Hospital, University of Zurich, from January 1, 1991 to December 31, 2014. The final diagnosis was based on the results of postmortem examination. The results were described in detail . The cowThe cows underwent a thorough clinical examination . GeneralThe following blood samples were collected from all cows: 5\u2009ml of EDTA blood for haematological analysis, 10\u2009ml of whole blood for serum biochemistry, 2\u2009ml of whole blood mixed with 0.2\u2009ml heparin for venous blood gas analysis and 5\u2009ml of EDTA blood for the glutaraldehyde test. Haematological analysis included the determination of haematocrit, total leukocyte count and the concentrations of fibrinogen and total protein using an automated blood analyser . The concentration of serum urea nitrogen was determined at 37\u2009\u00b0C using an automated analyser and the manufacturer\u2019s reagents (Roche Reagents) according to the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC). Venous blood gas analysis was done using an automated analyser . A glutaraldehyde test was done according to the manufacturer\u2019s instructions. Results were interpreted relative to reference intervals recently reported .A urine sample, mainly collected during spontaneous micturition, was analysed in 76 cows. The colour and transparency of the urine were assessed macroscopically, and the specific gravity was determined using a refractometer . A urine test strip was used to determine pH and the presence of protein, erythrocytes, glucose, ketones, leukocytes, nitrite, urobilinogen and bilirubin.A sample of rumen fluid (200 to 300\u2009ml) was collected using a Dirksen probe in 67 con\u2009=\u200958), abomasum (n\u2009=\u200921) and abdomen (n\u2009=\u200963) using a 3.5 or 5.0\u2009MHz convex or linear transducer [Seventy-five cows underwent ultrasonographic examination of the reticulum , 80\u2009mg/kg body weight intravenously.2, HCO3\u2212 and base excess of venous blood, urine pH, urine specific gravity). Differences in ulcer occurrence at various stages of lactation were analysed using a one-way analysis of variance and the post hoc Bonferroni test. A value of P\u2009<\u20090.05 was considered significant.The program IBM SPSS Statistics 22.0 was used for analysis. Frequencies were determined for each variable. The Wilk-Shapiro test was used to test the data for normality. Means \u00b1 standard deviations were calculated for normal data and medians for non-normal data , abdominal guarding (81%), congested scleral vessels (77%), ruminal atony (73%), tachycardia (68%), tachypnoea (65%), positive foreign body tests (58%), decreased skin surface temperature (53%), fever (49%), reduction in negative intraabdominal pressure assessed transrectally (39%), poorly digested plant fragments in faeces (35%) and arched back 28%) , spontaneous grunting and colic .All cows were ill and had reduced appetite or anorexia. Five (6%) cows were down at the time of admission, 24 (28%) were obtunded, 13 (15%) had muscle tremors and 24 (28%) had an arched back. Six (7%) cows had droopy ears and 4 (5%) had a droopy head. Two (2%) cows had a sawhorse stance and two others had abducted elbows. The principal signs of abdominal pain were bruxism (Table\u00a0Sixty-eight 81%) cows had abdominal guarding and six (7%) had abdominal distension. Rumen motility was reduced in 22 (26%) cows and absent in 63 (73%) , haemoconcentration (69%), azotaemia (56%), metabolic acidosis (49%), hyperfibrinogenaemia (45%), leukopenia 35%) and hypoproteinaemia (29%) in 35 (56%) cows and haematuria (5 to 250 erythrocytes per high-power field) in 33 44%) cows with macroscopically normal urine , left or right displacement of the abomasum (n\u2009=\u20097), fatty liver syndrome (n\u2009=\u20096), metritis or endometritis (n\u2009=\u20096), fascioliasis (n\u2009=\u20096), mastitis (n\u2009=\u20091) and ketosis (n\u2009=\u20091).Comorbidity occurred in 29 (33%) cows; 20 cows had one, seven cows had two and two cows had three or more additional diseases. The most common problems were claw disorders cows, and in 21 (24%) cows, the diagnosis was not clear. Fifty-four (62%) cows in which a type-4 ulcer was diagnosed were euthanased immediately except for a few that died during the examination. Twelve (14%) cows underwent right flank exploratory laparotomy to confirm the diagnosis and all were euthanased because of generalised peritonitis. The cows with an unclear diagnosis received a continuous intravenous infusion of a sodium-chloride-glucose solution , antibiotics and an NSAID , and two cows also received a magnet. Six cows died after the start of treatment and the remaining 15 were euthanased after two to four days because of deterioration in condition.All cows underwent portmortem examination and all had a type-4 abomasal ulcer and generalised peritonitis Figs.\u00a0 and 12. Forty-nine percent of all cases of type-4 abomasal ulcer occurred during the first four weeks of lactation, which had the highest incidence of all lactational stages. This was in general agreement with other reports , 19\u201321. Haematocrit and plasma protein are important laboratory variables in cows with type-4 ulcer. Increased haematocrit in 69% of cows indicated shock-induced haemoconcentration, but interestingly, this was accompanied by hyperproteinaemia in only 11% of cases. Usually an increase in haematocrit goes hand in hand with an increase in plasma protein concentration, but in the present study, 60% of cows had normal plasma protein concentrations and in 29% the concentration was even lower than normal. An increased haematocrit accompanied by normal or decreased plasma protein concentration is indicative of loss or active secretion of protein-rich fluid into the peritoneal cavity . It consAll but one of 47 peritoneal fluid samples had abnormalities typical of peritonitis including increased specific gravity and/or increased protein concentration, green discolouration suggesting contamination with ingesta or a putrid odour. However, it should be remembered that the standard classification of transudate and exudate do not always apply in sick cattle \u201342. In cUltrasonography was useful for visualising and assessing generalised peritonitis in 47 cows. The proximity of the abomasum to the reticulum showed reticular lesions in many cows with careful scanning of both sides. This underlines the importance of a thorough examination of the entire abdomen even in cases where lesions that support the tentative diagnosis are readily detected. For instance, when ultrasonographic examination is limited to the reticulum in cows with a history or clinical signs typical of traumatic reticuloperitonitis, the examiner runs the risk of missing changes associated with generalised peritonitis. Widespread peritonitic changes were seen in 26% of 503 cows with traumatic reticuloperitonitis . FibrinoCows with diffuse peritonitis attributable to a perforated abomasal ulcer respond poorly to treatment and repoThe diagnosis of type-4 abomasal ulcer based on clinical signs alone is difficult and therefore requires additional diagnostic procedures including the determination of the haematocrit and plasma protein concentration, abdominal ultrasonography and analysis of peritoneal fluid. In most cases, these steps lead to a correct diagnosis and allow timely euthanasia of the cow to prevent further suffering and unnecessary treatment costs. The dilemma faced by clinicians is that even though a correct diagnosis is usually possible, effective treatment of type-4 abomasal ulcer is not feasible. Efforts should therefore be made to minimise or eliminate ulcerogenic factors in cattle in later pregnancy and early lactation."} +{"text": "Proteobacteria isolated from Lechuguilla Cave in New Mexico. Genome-based phylogeny indicates that each strain belongs to a distinct genus. Two Rhizobiaceae isolates possess genomic potential for the biosynthesis of acyl-homoserine lactone.Genomic resources remain scarce for bacteria isolated from oligotrophic caves. We sequenced the genomes of five Proteobacteria isolated from Lechuguilla Cave in New Mexico. Genome-based phylogeny indicates that each strain belongs to a distinct genus. Two Rhizobiaceae isolates possess genomic potential for the biosynthesis of acyl-homoserine lactone.Genomic resources remain scarce for bacteria isolated from oligotrophic caves. We sequenced the genomes of five Rhizobiaceae isolates and confirmed the predicted phenotype using an Agrobacterium tumefaciens reporter assay . For the generation of genome-based phylogeny using GToTree v1.2.1 and LC148 (GenBank Protein accession numbers TKT46115 and TKT66962). The genes coding for these proteins were similarly classified as luxI homologs by antiSMASH 4 and the NCBI Prokaryotic Genome Annotation Pipeline (Agrobacterium tumefaciens NTL4(pZLR4) (retardation factor (fR) that is similar to that of 3-oxo-C8. LC148 produced two distinct AHLs, one with an fR value similar to that of C6 and the other with an fR value smaller than the included AHL standards (We used a previously described hidden Markov model (HMM) approach to ident4(pZLR4) . LC34 prtandards . Future tandards , targetetandards , or trantandards will be The raw Illumina paired-end reads and genome assemblies have been deposited in GenBank under the BioProject numbers listed in"} +{"text": "Mus musculus) adipocytes during insulin resistance progression, we uncovered the transcription factor Krueppel-like Factor 4 (KLF4) as a regulator of early insulin signaling. We experimentally confirmed that KLF4 controls the expression of two key insulin signaling molecules, the Insulin Receptor Substrate 2 (IRS2) and Tuberous Sclerosis Complex 2 (TSC2).One of the biggest challenges in analyzing high throughput omics data in biological studies is extracting information that is relevant to specific biological mechanisms of interest while simultaneously restricting the number of false positive findings. Due to random chances with numerous candidate targets and mechanisms, computational approaches often yield a large number of false positives that cannot easily be discerned from relevant biological findings without costly, and often infeasible, biological experiments. We here introduce and apply an integrative bioinformatics approach, Biologically Anchored Knowledge Expansion (BAKE), which uses sequential statistical analysis and literature mining to identify highly relevant network genes and effectively removes false positive findings. Applying BAKE to genomic expression data collected from mouse ( In silico network reconstruction techniques have been built upon quantitative measurements of relationships such as mutual information, pair-wise correlations, and conditional probabilities among interacting molecules i,random (i = 1 to 100), random distance matrices generated from random gene expression profiles. Random gene expression profiles were generated by permuting the observed gene expression profiles (Lanchor and Lneighbor genes) and repeating it 100 times. The reason for using permuted random gene expression profiles was to obtain a biologically relevant critical/threshold temperature at which real gene associations can be distinguished from random cluster formation. While the input for SPC clustering is the distance matrix, output is a temperature profile of clusters in the form of a matrix ([Y]) that provides the cluster membership of each gene at each temperature and chronicles the history of gene clusters from T = 0 to Tend. At T = 0 all the genes form one cluster. While the temperature is raised the large cluster gradually falls apart into smaller more stable sub-clusters until at Tend all clusters fall apart. The average (and standard deviation) of maximum cluster sizes at each temperature was estimated and compared between observed and random cases. Tcritical was determined as the lowest temperature at which the maximum cluster sizes for observed was significantly higher than for random cases , was converted to a \"visual representation\" of clusters by assigning a color to each cluster. A gradual shrinkage of clusters with rising clustering temperature can then be observed. This allowed us to identify strong associations between novel genes and anchor genes. Temperature profiles of clusters generated for anchor gene IRS2 are shown in We conducted SPC clustering on two sets of genes: 1) [X]om cases . StartinERT-Cre+/-/Klf4loxP/loxP) were generated as described in [ERT-Cre+/-/Klf4loxP/+) and wild type (ERT-Cre+/-/Klf4+/+) for the conditional allele were treated with tamoxifen (62.5 mg/kg body weight) in peanut oil by intraperitoneal injection starting at 6 weeks of age. Five days of treatment were followed by 2 days of rest and another 5 days of tamoxifen injections. At 12 weeks of age mice were euthanized. Parametrial and periovarian white adipose tissues were dissected, frozen in liquid nitrogen and stored at \u201380\u00b0C until RNA isolation. Total RNA was prepared from adipose tissues using TRIzol Reagent (Life Technologies Corporation) according to manufacturer\u2019s instructions. cDNA was synthesized from 2 microgram of total RNA using 1 micromolar oligo-dT (Life Technologies Corporation) and M-MLV reverse transcriptase (Life Technologies) in 20 microliter total volume following manufacturer\u2019s instructions. To the completed reaction 80 microliter H2O was added to obtain 100 microliter of diluted cDNA. Real time PCR was then performed on 6 microliter of the diluted cDNA (1X) using the qSTAR Expression Detection System (OriGene Technologies Inc) with IRS2 primers (Cat. No. MP206573), TSC2 primers (Cat. No. MP217683), KLF4 primers (Cat. No. MP207225) and MicroTubule-Associated Protein 1B primers , and the SensiMix SYBR Master Mix. MTAP1B was used as a negative control as its expression was not expected to be affected by changes in KLF4 expression. For PCR amplification the following conditions were used: initial denaturation and enzyme activation step at 95\u00b0C for 10 min followed by 42 cycles with denaturation at 95\u00b0C for 15 sec, annealing/extension/collection of data at 60\u00b0C for 60 sec. A melt curve analysis was performed for each reaction to confirm the amplification of single products. A standard curve for each primer set was obtained using different amounts of total cDNA input for one of the wild type samples and plotting the threshold cycles versus the log of cDNA input. Relative gene expression levels of IRS2, TSC2, KLF4, and MTAP1B in each sample were then derived from the standard curve.Tamoxifen-inducible KLF4-deficient mice (ribed in . For conIR+/-IRS1+/-ApoE-/- mice were fed a Western (high-fat) diet for 8 and 16 weeks starting at 8 weeks of age or a regular chow diet . Body we, n = 5) . Consistern diet . The lacent mice .0 genes) revealed that 33% played roles in cellular growth and proliferation and 31% in cell death and survival. Network enrichment analysis based on functional annotation of genes revealed genes belonging to gene networks associated with cellular assembly and organization, molecular transport, RNA trafficking, cell cycle, and inflammatory and infectious diseases.We analyzed genome-wide gene expression data to computationally discover genes that were differentially expressed in adipocytes of western and chow diet-fed mice. For this we simultaneously used two widely used statistical test procedures for microarray data analysis; LIMMA and SAM adjusted for multiple comparisons , 31. Whi0 genes) . Analysipath, Suppl. Table S3)(Step 2: Literature mining of known gene networks). To uncover novel genes linked to impaired insulin signaling, we then extracted genes from L0 that were strongly associated with the known insulin signaling pathway genes in Lpath (Step 3: Identification of novel genes associated with known network genes). Specifically, we correlated adipocyte gene expression patterns of L0 and Lpath for all the 26 animals in the different groups to capture possible functional associations across the different conditions representing type and duration of the diet (0 genes that were significantly correlated (Spearman\u2019s rank correlation \u03c1\u22650.72) with known insulin signaling pathway genes were considered candidate novel pathway genes in the insulin signaling pathway from the KEGG (Kyoto Encyclopedia of Genes and Genomes) database and the literature that showed significant expression changes between DW16 and DC16 with SAM test p-value <0.01 (1 genes that were most highly correlated with IRS2 as candidate novel network genes (LIRS2) (anchor) (top panel in We selected 15 anchor genes (represented by 20 Affimetrix probes), Lue <0.01 . Novel n (LIRS2) . PotentiTo further expand the insulin signaling pathway network, we determined the 20 closest neighbors around other anchor genes and performed SPC clustering with those and the 15 anchor genes (20 probes). We again examined tight clusters containing one or more anchor genes but not more than one novel gene. This analysis added novel genes to the network around anchor genes but also identified significant relationships between anchor genes . Among aBased on their interactions with different anchor genes five novel genes were introduced into the network: Chromodomain Helicase DNA binding protein 7 (CHD7), Amyloid beta (A4) Precursor-Like Protein 2 (APLP2), Ataxin-2 (ATXN2), and Kruppel-Like Factor 4 (KLF4) and REST CoRepressor 1 (RCOR1) . The infAmong the top 20 genes most highly correlated with IRS2 we extracted five genes that a literature search identified as transcriptional regulators: KLF4, ANKRD11, ZMYND8, CHD7 and NFIX . Among tKLF4; To reversely validate this novel gene\u2019s network interaction, we next expanded the network around KLF4 using the same strategy as described above for IRS2. From this reverse BAKE application, several known insulin signaling pathway genes including IRS2 were identified as close neighbors of KLF4 LKLF4; . In partKLF4+/-) [KLF4+/- mice compared to KLF4 wild-type mice . We founype mice . Thus, tpath adipog listed in anchor adipog) , EW16 (n = 8) and EC16 (n = 5)) to the data from IR+/-IRS+/-ApoE-/- mice when determining Spearman\u2019s rank correlation between gene expression profiles. Genes that were identified as novel after the application of the BAKE steps described above are listed in hidden, shown in bold in The results from the above analysis suggested a high prediction accuracy of BAKE for biological network inference. However, it was uncertain what degree of novel real network discovery could be achieved by the BAKE approach overall. Therefore, to evaluate overall accuracy of novel gene discovery and network relationships inferred by BAKE, we reconstructed an established adipogenesis regulatory network containing 84 pathway genes represented by 225 Affimetrix probes from the literature \u201351 and QNote that the performance values in We then also compared the performance of our adipogenesis network reconstruction inferred by BAKE with the most widely used and highly cited Network Inference (NI) approaches, the Gene Pattern-Dialogue on Reverse Engineering Assessment and Methods (GP-DREAM) , 24. GP-silico network mining techniques, however, often identify numerous false network interactions that can only be conclusively uncovered by performing extensive biological experiments. Some recent computational network inference techniques may yield as high as 98% false positives when identifying novel gene interactions in an adipogenesis network (Table 1). To overcome this critical shortcoming, we have developed BAKE, a new bioinformatics investigation strategy that is based on interactive sequential integration of known biological information and computational in silico analysis. Specifically, we have formalized an approach that researchers often already use to effectively investigate and validate their omics data-based discovery as five sequential BAKE analysis steps: in silico search \u2192 literature mining \u2192 integration and association of in silico search and literature mining \u2192 in silico network expansion around known network genes \u2192 in silico reverse-confirmation and use of literature and data resources for confirmation. Note that BAKE is an omics-based network inference approach rather than a fixed algorithm or software. Nevertheless, the full use and order of the five BAKE interactive and integrative analysis steps, combining computational discovery and known information, is important. In earlier BAKE attempts, we omitted some of these analysis steps and found that performance was significantly worse. For instance, gene clustering before biological knowledge anchoring resulted in numerous false positive gene clusters.Different computational network mining and modeling approaches have been developed in recent years to reconstruct complex biological networks from high throughput molecular data in genomics, proteomics, metabolomics and other omics-based studies. In IR+/-IRS1+/-ApoE-/-) may mediate down-regulation of IRS2 and TSC2 expression thereby impairing insulin signaling in adipocytes and promoting insulin resistance. To our knowledge, this is the first study to report the expression regulation of KLF4 on the key intermediates in the insulin signaling pathway, so a further investigation will be of high interest for its biological mechanisms and regulation.As we demonstrated, the implementation of the sequential BAKE analysis steps dramatically increased the likelihood of identifying true positives. When applying BAKE to an experimental animal model of insulin resistance progression, we discovered a novel regulatory transcription factor KLF4 to interact with IRS2 and TSC2, two key intermediates in the insulin signaling pathway. We were then able to experimentally corroborate a role for KLF4 in regulating IRS2 and TSC2 expression using KLF4 knockout mice. These investigations suggest that reduced expression of KLF4 triggered by a high-fat diet in our mouse model (neighbor + Lanchor = 40). Therefore, the number of novel genes (Lneighbor) that can be effectively introduced and computationally investigated with the known network genes is dependent on the number of anchor genes. For example, SPC with ~40 genes at K (number of nearest neighbors for clustering) = 10 generally produced stable clusters of 3 to 5 genes, which is an ideal size to study interactions between anchor and novel genes. If the total number was larger or smaller, the number or size of clusters tended to be too large or too small. BAKE/SPC algorithm is not directly sensitive to the number of genes in Lanchor but we used this empirical guideline for defining the number of novel genes to be investigated and suggest to keep Lanchor = Lneighbor = 20. This helps to generate more clusters involving anchor and novel genes, ideal for network expansion around known network genes in Step 4 of BAKE.One of the key features of BAKE was the use of SPC algorithm. The ability of SPC algorithm to naturally deduce gene relationships as well as their strength and closeness through the algorithm\u2019s temperature gradient without much intervention from the user was an attractive feature that led us to use it in BAKE. One of the questions that arose while implementing SPC algorithm in BAKE was to decide on the number of anchor and novel genes to be clustered by SPC. While it is difficult to define an exact number of anchor genes, we empirically found that our SPC and other algorithms we used perform well when the total number of both anchor and novel genes was around 40 and non-pathway genes (Lpathrandom) was repeated 100 times using Response Operator Characteristics (ROC) curves. Youden\u2019s J Index was estimated for the 100 comparisons in the ROC curve and the median correlation threshold for maximum Youden\u2019s J Index was estimated as 0.72. A more detailed description is provided under step 3 of BAKE in Materials and Methods.Comparison of correlation coefficients of L(PDF)Click here for additional data file.S2 FigThe central part of the temperature profile shown in (PDF)Click here for additional data file.S3 Fig\u00ae network analysis tool. Significant pathway genes ) are shown in green or red. Anchor genes (Lanchor adipog = 20 genes (20 probes)) are shown in red and hidden genes (Lhidden = 40 genes (53 probes)) in green. Eight genes were represented by different probes present in both the anchor and hidden gene list.The adipogenesis network was constructed based on literature \u201351 and Q(PDF)Click here for additional data file.S4 Figcritical (0.11 for the data shown) was defined as the lowest temperature at which maximum of cluster sizes was significantly higher for observed over random expression data or, in other words, the temperature beyond which the clustering pattern between random and observed gene expression data was distinct.Maximum cluster sizes at each temperature (shown as mean +/- SD estimated from N = 100 simulations of SPC clustering) were compared between temperature profiles of clusters obtained with observed gene expression data (red) and random gene expression data (green). T(PDF)Click here for additional data file.S1 TableParameters were determined in random-fed mice before or after euthanasia as described in Materials and Methods. Data are given as means +/- SEM.(PDF)Click here for additional data file.S2 Table0 represents genes that were differentially expressed between DW16 and DC16 adipocytes. L1 represents genes in L0 for which expression profiles significantly correlated with expression of insulin signaling pathway genes (Lpath) in adipocytes using data for all four conditions DC8, DW8, DC16 and DW16 (marked L1 in table).L(PDF)Click here for additional data file.S3 Tableanchor represents genes in Lpath that were differentially expressed in adipocytes between DW16 and DC16 (marked Lanchor in table). Fold Changes (FC) in gene expression between DW16 and DC16 are given in logarithmic scale (base 2).L(PDF)Click here for additional data file.S4 TableFold Changes (FC) in expression of neighbor genes between DW16 and DC16 are given in logarithmic scale (base 2).(PDF)Click here for additional data file.S5 TableNucleotide sequences 10 kb upstream of initiation start site ATG were scanned for KLF4 binding motifs ({G/A}{G/A}GG{C/T}G{C/T}) and the positions of motifs compared between human, mouse and rat promoters. Motifs shown in red were considered conserved based on the following criteria. They were found in the promoter of all three species, and located no more than 100 bases of each other across the different species and no more than ~ 1000 bases from the start site. Motifs that were less conserved across the three species due to single base variations are shown in green. Positions in promoters are given relative to translation start sites.(PDF)Click here for additional data file.S6 TableFold Changes (FC) in expression of neighbor genes between DW16 and DC16 are given in logarithmic scale (base 2).(PDF)Click here for additional data file.S7 TablePromoter regions of human, mouse and rat TSC2 were analyzed as described in legend to Suppl. Table S5. Positions in promoters are given relative to translation start sites.(PDF)Click here for additional data file.S8 Tablepath adipog, 52 genes represented by 73 Affymetrix probes) between DW16 and DC16 (p<0.01) are shown with fold changes in expression . Among these, 20 genes (represented by 20 Affymetrix probes) were randomly selected as anchor genes (Lanchor adipog). The remaining genes formed the list of hidden genes (Lhidden) to be discovered by BAKE through network expansion around anchor genes.List of differentially expressed adipogenesis network genes (L(PDF)Click here for additional data file.S9 TableGenes are listed in alphabetical order. Hidden adipogenesis network genes discovered among novel genes are in bold.(PDF)Click here for additional data file.S10 Table\u00ae). In IPA gene interactions are derived and curated from a variety of databases such as Ingenuity Expert Information, microRNA-mRNA interaction database (miRecords), protein-protein interaction databases , BioGRID, Gene Ontology (GO), Online Mendelian Inheritance in Man (OMIM) and Mouse Genome Database (MGD).Anchor gene-anchor gene and anchor gene-hidden gene interactions inferred by BAKE are shown along with their classification as True Positives (TPs) and False Positives (FPs). Literature evidence for TP gene interactions were obtained from the adipogenesis network shown in (PDF)Click here for additional data file."} +{"text": "TP53 (49%), followed by KRAS (31%) and EGFR (13%); in mCRC, TP53 (50%), KRAS (48%) and PIK3CA (16%) were the most frequently mutated genes. Moreover, NGS identified actionable genetic alterations in 58% of NSCLC patients, and 49% of mCRC patients did not harbor primary resistance mechanisms to anti-EGFR treatment. Validation with conventional approaches showed an overall agreement >90%. Turnaround time and cost analysis revealed that NGS implementation is feasible in the public healthcare context. Therefore, NGS is a multiplexed molecular diagnostic tool able to overcome the limitations of current molecular diagnosis in advanced cancer, allowing an improved and economically sustainable molecular profiling.The establishment of precision medicine in cancer patients requires the study of several biomarkers. Single-gene testing approaches are limited by sample availability and turnaround time. Next generation sequencing (NGS) provides an alternative for detecting genetic alterations in several genes with low sample requirements. Here we show the implementation to routine diagnostics of a NGS assay under International Organization for Standardization (UNE-EN ISO 15189:2013) accreditation. For this purpose, 106 non-small cell lung cancer (NSCLC) and 102 metastatic colorectal cancer (mCRC) specimens were selected for NGS analysis with Oncomine Solid Tumor (ThermoFisher). In NSCLC the most prevalently mutated gene was Cancer is a complex and heterogeneous disease with considerable variation in histological and biological features. Understanding the role of genetic alterations involved in cancer development has led to its reclassification into different molecular subtypes that reflect biological behavior and may lead to further effective therapeutic targets to achieve improved outcome ,2. For tThe knowledge of a tumor\u2019s genetic profile is crucial to improve clinical-decision making in the patient management. Consequently, laboratories must integrate high-throughput sequencing technologies in routine molecular diagnostics . These aThe purpose of the current study is to evaluate the integration of NGS technology in a routine clinical setting. We describe the mutational profile of two highly prevalent cancers (advanced NSCLC and mCRC) and analyze its diagnostic potential to characterize molecular heterogeneity and to increase the therapeutic opportunities with targeted therapies; we assess NGS technology at a technical and economical level; and we describe our experience in clinical practice of an NGS pipeline for cancer molecular diagnostics in the UNE-EN ISO 15189:2013 accreditation scope.EGFR p.Leu858Arg and KRAS p.Gly13Asp mutations in which NGS VAF was slightly higher .NGS assay was able to detect the seven low frequency variants (between 1\u20133%) present in two reference materials used as positive controls. The variant allele frequency (VAF) detected was consistent to the data obtained by digital droplet PCR (ddPCR) assays except for Quality control analysis revealed excellent performance of the NGS panels . The medOn the other hand, NGS showed an invalid test rate of 3.8% (8/208 FFPE specimens). Six NSCLC samples and two mCRC failed due to low sequencing quality metrics . These samples were subsequently excluded from the study.ROS1 rearrangements and NRAS mutations the overall agreement (OA) was 100% . Instead, NGS and Sanger Sequencing (SS) revealed a synonymous change in homozygosis in 787 codon p.(Gln787Gln). NGS technology allowed the detection of an ALK rearrangement not detected by immunohistochemistry (IHQ) or fluorescence in situ hybridization (FISH) (OA: 99.0%). In KRAS, seven specimens gave discordant results when comparing with Real Time (RT)-qPCR assay (OA: 94.7%). BRAF pVal600Glu mutation was detected by High Resolution Melting (HRM) in nine out of eleven NGS BRAF p.Val600Glu mutated samples (OA: 96.4%).For was 100% . RegardiEGFR, ALK and ROS1) and mCRC . Starting in both cases from FFPE tissue blocks we were able to prepare libraries, sequence eight NSCLC or ten mCRC samples, and analyze data in five working days. Conventional methodologies for molecular testing of EGFR, ALK and ROS1 resulted approximately in three days while testing KRAS, NRAS and BRAF in mCRC resulted approximately in four working days. However, if KRAS is positive, TAT is reduced to three days. TAT and cost comparison between NGS and conventional methods is shown in In order to compare NGS with conventional methodologies under theoretical conditions, we calculated the turnaround time and cost for three mandatory testing genes in NSCLC . Moreover, this NGS assay was externally validated within the European Genetics Quality Network (EMQN) External Quality Assessment Scheme for Oncogene Panel Testing, obtaining satisfactory results in 2017 and 2018 editions [Sequencing analysis identified on average 1.45 non-synonymous and non-polymorphic variants per sample (291/200) . After fTP53 (49%) followed by KRAS (31%), EGFR (13%), BRAF (11%) and PIK3CA (7%). Rearrangements were found in ALK and ROS1 . In the entire group, 9% of patients did not carry any somatic mutation; 56% harbored one somatic mutation and 35% two or more . Mutations in KRAS were the second most prevalent (48%), specifically mutations in codon 12 accounted for 37%. We also detected pathogenic variants in codon 13 (5%), codon 146 (5%), codon 117 (1%) and the uncommon codon 19 mutation (p.Leu19Phe) found in concomitancy with a codon 146 mutation. Pathogenic variants in PIK3CA supposed 16% and mutations in SMAD4 were detected in 11% of patients. Regarding BRAF, eight samples harbored the classical p.Val600Glu and two showed mutations outside this hotspot. NRAS mutated samples (2%) harbored the hotspot p.Gln61Arg mutation , PIK3CA (n = 5), SMAD4 (n = 3) and FBXW7 (n = 1), CTNNB1 (n = 1) and AKT (n = 1). BRAF p.Val600Glu mutation was found in concurrency with TP53 (n = 3), PTEN (n = 1), PIK3CA (n = 1) and SMAD4 (n = 1). NRAS and PIK3CA were concomitant in one sample. Fifteen patients carried three concurrent mutations, KRAS-PIK3CA-TP53 being the most frequent combination (n = 5). Interestingly, one patient harbored concurrent mutations in KRAS, BRAF and TP53 and other carried mutations in NRAS, BRAF and TP53. One patient carried four concurrent mutations in KRAS, SMAD4, FBXW7 and TP53 (Fifty mCRC patients harbored concurrent mutations. and TP53 .EGFR were exon 21 mutations (6%) followed by exon 19 alterations (5%). Codon 12 was the most frequently mutated in KRAS (23%) followed by codon 13 (3%) and codon 61 (2%). In NRAS only codon 61 was found mutated (1%). Regarding BRAF, three out eleven detected mutations occurred on the hotspot Val600. In regard to PIK3CA, codon 542 mutations were the most frequent (2%) followed by mutations in codons 545 and 1047 (1% for both). Duplication in exon 20 and the hotspot mutation p.Arg784His (1%) were found in ERBB2. Four patients showed fusions between ALK and EML4, in all of them, the rearrangement involved exon 20 of ALK, in three patients with exon 6 of EML4 and in other with exon 13. One patient showed a fusion of ALK with an unknown partner. Finally, one patient presented a fusion between ROS1 (exon 35) and CD74 (exon 6).NGS identified actionable genomic alterations in 58% of NSCLC patients . The mosRAS mutations as a primary resistance mechanism to anti-EGFR therapies. Additionally, in the RAS wild type patients, NGS identified eight BRAF V600E mutated patients, five PIK3CA mutated patients and one patient harboring p.Lys57Asn in MAP2K1 gene . However, since both NGS and SS did not detect this mutation, we hypothesize that the synonymous variant could affect primer or probe hybridization of the Cobas\u00ae assay, resulting in a false positive detection of the p.Thr790Met mutation. In KRAS testing, we found seven discordant cases. Two samples resulted positive by RT-qPCR assay but were not detected by NGS. These samples were re-tested using a new lot of the AmoyDx assay, providing then concordant results with the NGS assay. Among the five negative samples for KRAS mutations by RT-qPCR, NGS reported mutations at low VAF in two cases . In theory, these VAFs should be detected by the AmoyDx assay, which has a limit of detection (LOD) of 1\u20132%, established by using cell line DNA. However, we suspect this LOD could be higher when using highly degraded DNA obtained from FFPE samples. In the three remaining cases, NGS revealed KRAS mutations at high VAF , which could be confirmed by SS. Moreover, these samples were re-tested by a technician in another institution, showing concordant results with the NGS assay. For BRAF p.Val600Glu mutation, NGS revealed two mutated samples with VAF of 4% and 5%, not detected by HRM (LOD = 10%). Regarding fusion transcripts, an OncoNetwork collaborative research study was able to detect EML4/ALK fusion up to 1% dilution [Moreover, we found an excellent correlation between NGS and single-gene conventional methods. The analysis of discordant results revealed that NGS is a more robust method compared with conventional approaches. Concerning dilution . In our dilution . Taken tdilution .KRAS mCRC patients. In KRAS mutated patients this benefit is not observed, however, a complete NGS test is achieved with a \u20ac30 difference per sample, therefore being an economically sustainable approach. In NSCLC, the extra cost associated with NGS studies (\u20ac51.5/patient) can be assumed based on the ability to identify actionable alterations with significant impact on patients\u2019 outcome. TAT and economic costs are essential for NGS implementation in routine molecular diagnostics in a public healthcare hospital. Here, we found a great economic benefit in the employment of NGS technology versus conventional methodologies when it came to wild type KRAS mutational status). This delay in molecular studies should not be an important limitation of NGS implementation because of its ability to identify clinically relevant alterations beyond the routinely tested genes. Moreover, the coexistence of both strategies may allow the choice of a faster conventional strategy when needed, especially in patients whose clinical situation requires a molecular result in a short period of time.The NGS approach described in this study requires a manual library and template preparation . Consequently, hands-on time is clearly higher than conventional studies in both NSCLC and mCRC samples. However, the development of new automatized devices for library and template preparation has drastically reduced hands-on time to approximately 1 hour, making NGS implementation in terms of technical staff much easier. Global time duration of NGS studies has also been higher than conventional approaches in NSCLC (5 versus 3 working days) and in mCRC and automatizing the process (IonChef Instrument). Moreover, the development of new and faster sequencers (Ion S5 Instrument) is able to reduce global time duration to 4 days. However, it is important to acknowledge that NGS is economically sustainable when the appropriate number of samples is studied in the same experiment. In this sense, and according to the number of samples received for NGS studies, we are reporting NGS results under routine laboratory conditions in approximately 10\u201315 working days, as recommended ,29,30.The major advantage of the NGS approach is to provide information about potential therapeutic targets to improve clinical outcomes of patients with advanced cancer. Multiple biomarker testing has become a major challenge for molecular diagnostic laboratories because of the increasing number of approved targeted therapies and clinical trials. In this scenario NGS has been postulated as a technology with clinical applicability able to provide an exhaustive molecular profiling, deciphering tumoral heterogeneity that can in certain cases have a prognostic value and/or explain treatment resistance .RAS wild type patients (n = 50) NGS identified four patients with mutated PIK3CA. Response to anti-EGFR treatment in these patients is still controversial [RAS-RAF and PIK3CA wild-type patients seem to have better responses [MAP2K1 that has been described as a primary resistance mechanism to anti-EGFR treatment [The mutation prevalence identified in our study for NSCLC and mCRC samples is concordant with previously published studies ,33,34. Eoversial althoughesponses . Moreovereatment . Taken tTP53 [STK11 in concomitancy with KRAS mutations [TP53 [SMAD4 [FBXW7 [Concurrent mutations have been detected in 35% of NSCLC patients and in 50% of mCRC patients revealing tumor biology complexity. Although there are no well-established molecular prognostic factors neither in NSCLC nor mCRC certain passenger mutations may be associated with an adverse prognosis. In this sense, TP53 or STK11utations have beens [TP53 or SMAD43 [SMAD4 mutation4 [FBXW7 has receThe study included a series of 106 advanced NSCLC (stages III\u2013IV) and 102 mCRC (stage IV) patients diagnosed in the Department of Medical Oncology at the University Hospital La Fe from 2015 to 2017. The epidemiological, clinical and pathological features of these patients are summarized in Genomic DNA was isolated from three 5 \u03bcm thick FFPE sections using Deparaffinization Solution and the GeneRead DNA FFPE Kit . RNA was extracted from three 15 \u03bcm thick FFPE sections employing the RecoverAllTM Total Nucleic Acid Isolation Kit . DNA and RNA concentration was assessed using Qubit 3.0 fluorometer with DNA HS or RNA HS Assay Kit (ThermoFisher Scientific).Conformit\u00e9 Europ\u00e9enne-In vitro diagnostic (CE-IVD) approved kits and workflows.Molecular analysis was performed at the Molecular Biology Unit using AKT1 (NM_001014431.1), ALK (NM_004304.4), BRAF (NM_004333.4), CTNNB1 (NM_001904.3), DDR2 (NM_006182.2), EGFR (NM_005228.3), ERBB2 (NM_004448.3), ERBB4 (NM_005235.2), FBXW7 (NM_033632.3), FGFR1 (NM_001174067.1), FGFR2 (NM_022970.3), FGFR3 (NM_001163213.1), KRAS (NM_033360.3), MAP2K1 (NM_002755.3), MET (NM_001127500.1), NOTCH1 (NM_017617.3), NRAS (NM_002524.4), PIK3CA (NM_006218.2), PTEN (NM_000314.4), SMAD4 (NM_005359.5), STK11 (NM_000455.4), TP53 (NM_000546.5)). The design includes 92 amplicons. For RNA sequencing of NSCLC samples, we used Oncomine Solid Tumor Fusion Transcript kit , that allows the detection of fusion transcripts involving ALK, RET, ROS1 and NTRK1 genes with 85 amplicons. All NGS studies were conducted with the Ion Torrent Personal Genome Machine (PGM) technology (ThermoFisher Scientific).Oncomine Solid Tumor DNA kit was used for mutation detection in 22 genes involved in colon and lung cancer . Finally, quantification and dilution (100 pM) of the amplified libraries was performed using the Ion Library Equalizer Kit (ThermoFisher Scientific) as described by the manufacturer. For DNA libraries preparation, multiplex PCR was performed on 10 ng of DNA. After primer digestion and barcode ligation, library fragments were purified with AgencourtRNA libraries preparation included a previous cDNA synthesis step from 10 ng of RNA using the SuperScript kit VILO cDNA synthesis kit (ThermoFisher Scientific). In this case, a multiplex PCR amplification of cDNA was performed. Library quantification was carried out by qPCR, inferring the concentration from a standard curve generated with Ion Library Quantification Kit (ThermoFisher Scientific). RNA libraries were diluted to a concentration of 100 pM.In NSCLC, DNA and RNA libraries from eight patients were combined in a 4:1 proportion, generating the library pool. In mCRC samples, 10 DNA libraries were combined in equal proportion.The library pool was clonally amplified in an emulsion PCR reaction using Ion Sphere Particles (ISPs) in the One Touch 2 Instrument. Subsequently, template-positive ISPs were enriched using the Ion One Touch ES with the Ion PGM Hi-Q OT2 kit following manufacturer\u00b4s protocol. Enriched template-positive ISPs were subjected to sequencing on the Ion Torrent Personal Genome Machine (PGM) on a 318v2 Ion Chip using Ion PGM Sequencing Hi-Q kit . Raw data processing and alignment to the hg19 human reference genome was performed with Torrent Suite v5.6. Aligned sequences (Binary Alignment Map (BAM) files) were automatically transferred to the Ion Reporter Software (v5.6) to perform variant calling/annotation by using commercial workflows. Intronic variants and synonymous changes were filtered out. Variants with low total read depth <500 total) and/or low variant read depth (<20 reads) were excluded. Additionally, Variants were visually examined using the Integrative Genomics Viewer (IGV) software (v.2.4). Subsequently, sequence variation databases such as Catalogue of Somatic Mutations in Cancer (COSMIC) , VarSome0 total aVerification instead of a full validation analysis was performed according to our national accreditation body , since the OST-DNA and OST-RNA kits are CE-IVD approved. The performance of NGS testing was extensively evaluated on different aspects. Firstly, we used well-characterized reference material to assess the presence or absence of somatic variants and their allele frequencies. Secondly, we considered the pre-analytical conditions and assessed the quality of NGS analysis on determining FFPE samples as start material, allowing us to establish the sequencing quality metrics. Thirdly, diagnostic sensitivity and specificity were determined experimentally by comparing it with conventional methods for routinely tested alterations; additionally, clinical reporting was adapted according to international diagnostic standards and professional guidelines. Finally, to ensure a consistent high standard of performance, it was essential to establish an EQA program to monitor the quality of NGS testing in clinical practice and to propose corrective actions when needed.EGFR hotspot mutation p.Leu858Arg (VAF:3%), the EGFR deletion p.Glu746_Ala750del (VAF:2%) and the resistance hotspot mutation p.Thr790Met (VAF:1%) and the other harbored the following low frequency variants: EGFR p.Leu858Arg (VAF:3%), p.Thr790Met (VAF:2%), KRAS p.Gly13Asp (VAF:3%) and PIK3CA p.His1047Arg (VAF:3%). All described variants had previously been validated by ddPCR. To evaluate the performance of the NGS assay for low frequency variant (<5%) detection we used reference materials provided by the European Molecular Genetics Quality Network (EMQN) in the External Quality Assessment Scheme for Oncogene Panel Testing (2017 and 2018). One of the reference materials used harbored the The number of reads, mean depth, \u201con-target\u201d reads and uniformity were the parameters used as quality control check points for further sample analysis. A total number of reads higher than 100,000 together with \u201con-target\u201d and uniformity values >80% were required for each DNA library and 20,000 total reads for each RNA library.EGFR, NRAS, KRAS and BRAF genes, as well as ALK and ROS1 genes rearrangements were tested by conventional methods. EGFR mutations were validated by Cobas\u00aeEGFR Mutation Test v2 (CE-IVD) ; NRAS and KRAS mutations were confirmed by Real Time (RT)-qPCR using AmoyDx\u00aeKRAS Mutation Detection Kit and AmoyDx\u00aeNRAS Mutation Detection Kit ; BRAF mutations were validated by High Resolution Melting (HRM) as previously described [ALK and ROS1 rearrangements were studied by immunohistochemistry (IHQ) employing VENTANA ALK (D5F3) CDx Assay and IHQ ROS1 Clon D4D6 , respectively. Positive IHQ assays were confirmed by fluorescence in situ hybridization (FISH) employing Vysis ALK Break Apart FISH Probe Kit and Vysis 6q22 ROS1 Break Apart FISH Probe Kit (Abbott Laboratories), respectively.Detected missense mutations in escribed . ALK andTo ensure a consistent high standard of performance, this assay was externally validated by the participation in the EMQN External Quality Assessment Scheme for Oncogene Panel Testing (2017/2018).Quantitative variables were summarized by their mean and standard deviation, and categorical variables by absolute frequencies. NGS is a technology able to assess multiple genetic biomarkers that has demonstrated a great concordance with conventional single target assays, providing an exhaustive molecular profiling of clinically relevant alterations at reasonable costs and turnaround times. The implementation of NGS in the diagnostic routine under the scope of UNE-EN ISO 15189:2013 accreditation has provided relevant information for patients\u2019 clinical management, improving the molecular diagnostic in our center."} +{"text": "Mechanical thrombectomy (MT) using stent retriever assisted vacuum-locked extraction (SAVE) is a promising method for anterior circulation strokes. We present our experience with SAVE for large vessel occlusions (LVO) of the posterior circulation.We retrospectively analyzed 66 consecutive MT patients suffering from LVO of the posterior circulation. Primary endpoints were first-pass and overall complete/near complete reperfusion, defined as a modified thrombolysis in cerebral infarction (mTICI) score of 2c and 3. Secondary endpoints contained number of passes, time interval from groin puncture to reperfusion and rate of postinterventional symptomatic intracranial hemorrhage (sICH).p\u2009=\u20090.0249). Median groin to reperfusion time did not differ significantly between groups. The rate of sICH was 5% without any complications in the SAVE cohort.Median age was 75\u2009years (interquartile range (IQR) 54\u201381\u2009years). Baseline median National Institutes of Health stroke scale (NIHSS) was 13 (IQR 8\u201321). Fifty-five (83%) patients had LVO of the basilar artery and 11 (17%) of the posterior cerebral artery. Eighteen (27%) patients were treated with SAVE and 21 (32%) with aspiration only. First pass mTICI2c or 3 and overall mTICI2c or 3 were documented in 11/18 (61%) and 14/18 (78%) with SAVE and in 4/21 (19%) and 13/21 (33%) with aspiration only. Median attempt was 1 (IQR 1\u20132) with SAVE and 2 (IQR 1\u20134) with aspiration (Mechanical thrombectomy of posterior large vessel occlusions with SAVE is feasible, safe, and effective with high rates of near-complete and complete reperfusion. Mechanical thrombectomy (MT) is the standard treatment for patients suffering from acute ischemic stroke (AIS) due to intracranial large vessel occlusion (LVO) in the anterior circulation since large randomized controlled trials (RCT) showed efficacy of this strategy over standard medical care . The timIn recent years, thrombectomy techniques have evolved rapidly by advances in catheter technology in order to improve the rate of successful reperfusion and minimize occurrence of distal embolization. The main methods of MT for LVO include stent retriever thrombectomy with or without the use of an aspiration catheter or the uWe report a single-center, retrospective analysis of patients, who were treated with MT for AIS caused by LVO in the posterior circulation from March 2014 to November 2018 in our department. Baseline parameters, angiographic features, technical and clinical outcome were derived from a prospectively maintained neurointerventional database. Primary endpoints were first pass and overall complete or near-complete reperfusion, defined as the modified Thrombolysis in Cerebral Infarction (mTICI) scale score of 2c and 3 [Following inclusion criteria were applied: clinical diagnosis of AIS due to LVO in the posterior circulation and initiation of endovascular stroke treatment with complete angiographic documentation. There were no general limitations on baseline variables such as age and National Institutes of Health Stroke Scale (NIHSS) on admission or procedural characteristics, including the use of different thrombectomy techniques and intra-arterial thrombolysis, which were left to the attending neuroradiologist\u2019s discretion. According to neurological guidelines, patients received IVT whenever possible. Procedural angiograms were reviewed independently by experienced interventional neuroradiologists to evaluate the mTICI score before and after MT and blinded to the used MT technique. In accordance to the anterior circulation, reperfusion was understood as contrast material passes beyond the area of initial occlusion. Therefore, the territory distal to the occlusion was set as 100% and percent reperfusion was measured and translated into the mTICI scoring system. Ischemic stroke was confirmed with native computed tomography (CT) or flat panel detector CT in cases of one stop management and CTA (or multiphase FDCTA) . Native The SAVE technique has been recently described . Brieflyhttp://www.medcalc.org; 2019). Descriptive statistics of normally distributed data are stated as mean and standard deviation; not normally distributed data are summarized as median and interquartile range (IQR). Differences between groups were examined using Fisher\u2019s exact test or Mann-Whitney test. Statistical significance was defined as p\u2009\u2264\u20090.05.All analyses were conducted using MedCalc Statistical Software version 18 were male and median age was 75\u2009years IQR 54\u201381\u2009years). Median baseline NIHSS was 13 (IQR 8\u201321) and the proportion of patients receiving IVT was 56% . The rate of first pass successful reperfusion (mTICI \u22652b) was also higher in the SAVE group with 12/18 (67%) vs. aspiration group with 7/21 . An overall rate of mTICI 2c and 3 on final angiogram was achieved in 14/18 (78%) SAVE cases and in 7/21 (33%) aspiration cases (p\u2009=\u20090.0061). A total of 12/18 (67%) SAVE patients and 3/21 (14%) aspiration patients were completely reperfused (mTICI 3) at the end of the procedure (p\u2009=\u20090.0009). The overall rate of successful reperfusion (mTICI \u22652b) was 16/18 (89%) with SAVE vs. 13/21 (62%) with aspiration. Median attempt was 1 (IQR 1\u20132) with SAVE and 2 (IQR 1\u20134) with aspiration (p\u2009=\u20090.0249). Median groin to reperfusion time did not differ significantly between SAVE and aspiration only groups (46\u2009min (IQR 29\u201356) vs. 39\u2009min (IQR 22\u2013102); p\u2009=\u20090.9723).The primary endpoint of first pass complete or near complete reperfusion mTICI \u22652c) was achieved in 11/18 (61%) with SAVE vs. 4/21 (19%) with aspiration (c was achNo complications occurred with SAVE. Two patients (5%) receiving aspiration only suffered from sICH. There were no statistical differences observed in median NIHSS and mRS at discharge between the two groups: median NIHSS score at discharge was 5 for both groups and favorable neurological outcome (mRS\u2009\u2264\u20092) was confirmed in 6/18 (33%) patients treated with SAVE and 8/21 (38%) patients treated with aspiration only. Overall, favorable outcome at discharge was 24/66 (36%) and median NIHSS score at discharge was 5 (IQR 2\u201312).So far, there is one randomized trial, which compared the safety and efficacy of MT plus standard medical therapy vs. standard medical therapy in patients suffering from posterior circulation strokes . The BESAs the central requirement of MT is a fast recanalization of the occlusion with complete reperfusion of the affected territory, new devices and different strategies were developed during the last years. Promising techniques using large-bore aspiration catheters and/or new generation stent retrievers had shown promising results with high reperfusion rates, however, those techniques were mainly applied in the anterior circulation , 19. ForNew aspiration catheters and aspiration pumps have been implemented with different results in achieving high and constant flows , 26. In Although the procedure times by using SAVE tended to be longer compared to aspiration only, no differences in clinical outcome at discharge were observed, which might possibly be explained by the better reperfusion results. However, as stated in the literature up to now , this stEffectiveness of proximal flow arrest/aspiration in the posterior circulation is matter of debate and limited to few cases , 28. WhiA limitation of our study is the retrospective design with the attendant selection bias. The small sample size limits the validity of the data. The observed differences between the techniques should be interpreted with caution as in our institute experience with SAVE is high compared to the aspiration only technique. The small sample size and missing evaluation of the aforementioned anatomic factors might prevent the conclusion that SAVE is more effective than aspiration. Clinical outcome after 90\u2009days is missing; however, our intention was to focus on the angiographic results as complete reperfusion is a basic requirement for recovery of stroke patients. A prospective trial is warranted to demonstrate efficacy of SAVE for posterior circulation strokes.Mechanical thrombectomy of posterior large vessel occlusions with SAVE is feasible, safe, and effective with high rates of near-complete and complete reperfusion."} +{"text": "Mycobacterium abscessus (2/12) co\u2010infection. Antibiotics, systemic corticosteroids, and therapeutic lavage were interventions in all eight and five patients, respectively. Clinicians should consider ELP in children with non\u2010resolving pneumonia in settings with similar practices.Exogenous lipoid pneumonia (ELP), an important cause of interstitial lung disease, often goes unrecognized. We conducted a retrospective study of children with histologically confirmed ELP at Red Cross Children\u2019s Hospital, South Africa. Twelve children of Zimbabwean heritage aged 2.1\u201310.8 months were identified between 2012 and 2017. Repeated oral administration of plant\u2010based oil for cultural reasons was reported by 10 of 11 caregivers. Cough (12/12), tachypnoea (11/12), hypoxia (9/12), and diffuse alveolar infiltrates on chest radiography (12/12) were common at presentation. Chest computed tomography revealed ground\u2010glass opacification with lower zone predominance (9/9) and interlobular septal thickening (8/9). Bronchoalveolar lavage specimens appeared cloudy/milky, with abundant lipid\u2010laden macrophages and extracellular lipid on Oil\u2010Red\u2010O staining (12/12), with polymicrobial (6/12) and Exogenous lipoid pneumonia (ELP) is considered a rare disorder caused by inhalation or aspiration of mineral, plant\u2010based, or animal oils Oil administration may be a common practice that is not recognized to be potentially dangerous by caregivers or medical professionals. This history may not be forthcoming from caregivers unless health workers probe for it explicitly, leading to misdiagnosis, treatment delay, and a missed opportunity to prevent ongoing oil aspiration , we conducted a retrospective case series at Red Cross Children\u2019s Hospital, a tertiary referral hospital in South Africa.Following ethical approval from the University of Cape Town Human Research Ethics CommitteeWe consecutively selected children aged <18 years with histologically confirmed ELP defined under the following conditions: (1) clinical presentation of persistent or recurrent unexplained pneumonia associated with tachypnoea or hypoxia; (2) radiological evidence of persistent diffuse alveolar infiltrates on chest radiography or chest computed tomography (CT); (3) extracellular lipid and lipid\u2010laden macrophages in bronchoalveolar lavage (BAL) and/or frozen section lung biopsy; and (4) history of oil administration related to time of presentation/diagnosis.Permission to search patient records including medical folders and contact information was obtained from the hospital management. The lead researcher reviewed medical records of eligible patients and extracted relevant clinical data. Data were captured electronically, anonymized, and recorded in a standard case report form. The study radiologist and pathologist independently reviewed and entered data related to chest radiography, CT findings, BAL, and lung biopsy findings.Children with suspected interstitial lung disease (ILD) routinely underwent high\u2010resolution CT utilizing a paediatric\u2010friendly radiation dosing protocol with controlled ventilation if under the age of six years. Flexible bronchoscopy (Olympus\u00ae 2.8 mm) and radiology\u2010directed BAL was performed in all children under general anaesthesia through a laryngeal mask. Histocytological assessment of BAL and lung biopsy specimens routinely include Oil Red O stain for lipids, periodic acid Schiff stain for glycoprotein exudates, Perls\u2019 Prussian Blue stain for iron, Grocott\u2019s methanamine silver stain for fungi, and Ziehl\u2010Neelsen stain for acid fast bacilli. Processing and examination of BAL and lung tissue were conducted according to the European Management Platform for Childhood Interstitial Lung Disease protocols Between October 2012 and December 2017, we identified 12 children with ELP as per our study case definition. All children were of Zimbabwean heritage, presenting in infancy (range 2.1\u201310.8). Dry cough was the main presenting symptom in all children with a duration varying from one day to three months. Common symptoms at presentation of these infants included: tachypnoea (11/12), hypoxia (9/12), fever (4/12), hyperinflation (2/12), and digital clubbing (2/12). Six of 12 children were hospitalized for pneumonia on at least one other occasion besides the episode in which the diagnostic BAL that was conducted provided histocytological evidence of ELP. Underlying risk factors that were documented included gastro\u2010oesophageal reflux (GOR) confirmed on scintigraphy (3/6), in combination with silent aspiration confirmed on contrast swallow (2/4). Ten of the 11 mothers interviewed confirmed the administration of oil to their children. In five patients, this history was obtained after reviewing BAL findings. Oil administration emerged to be a nearly universal cultural practice by Zimbabweans, even in the Cape Town diaspora. ground\u2010glass opacification with lower zone predominance (9/9); (2) smooth interlobular septal thickening (8/9)/crazy paving appearance (5/9); (3) expansile right upper lobe consolidation (2/9); and (4) fat attenuation within the areas of airspace consolidation (1/9). These patterns were present in various combinations.Twelve BAL samples and one frozen section lung biopsy were assessed. On gross inspection, all BAL specimens were cloudy, 11 of 12 being predominantly milky in nature ; oxygen supplementation (11/12), non\u2010invasive ventilation (8/12), and mechanical ventilation (1/12); antibiotics (12/12); and systemic corticosteroids (8/12). Five patients underwent partial therapeutic lung lavage. Four children needed only one procedure to obtain satisfactory clinical response, and one needed three sequential lavages over 11 weeks before achieving satisfactory clinical improvement. The median hospital stay duration was 23 (range 2\u2013117) days. Clinical resolution was documented in 10 of 12, with a median time to clinical resolution from presentation of 1.1 (range 0.3\u201314) months. To date, radiological resolution on plain radiography from presentation is only documented in two, at 19.4 and 27.0 months, respectively. No mortalities were reported, and the children continue to be followed up in our service.Exogenous lipoid pneumonia in children has been described in Asia, the Americas, and Europe M. abscessus co\u2010infection has not previously been reported in children with ELP. Other Nontuberculous Mycobacteria (NTM) described in children with ELP include Mycobacterium fortuitum, Mycobacterium chelonei, and Mycobacterium smegmatisMycobacterium abscessus complex has been reported in adult patients with ELP Furthermore, M. abscessus disease. This possibly reflects a severe form of lung oleoma/paraffinoma characteristically seen in adults Diffuse ground\u2010glass opacification, predominantly in the posterior segments, and interlobular thickening were common patterns on chest CT in children with ELP consistent with the literature. Fatty attenuation within consolidation has also been described in children with ELP; however, we only observed this pattern in one child Discontinuing oil, treating infections, identifying underlying risk factors, and overall supportive care in line with consensus reviews In conclusion, our case series highlights that ELP is an uncommon but serious condition that may occur in any global context with similar cultural practices. Obtaining a history of oil administration from caregivers, chest radiography and cytological analysis of BAL are sufficient to diagnose ELP. NTM co\u2010infection should be excluded in children with suspected ELP. Health education messages to highlight the risks associated with the cultural practice of oil administration are needed.This study was approved by the University of Cape Town Human Research Ethics Committee (548/2017)."} +{"text": "HFE gene variants has been studied in many cancer types; however, the impact of HFE variants, sex and HFE gene expression in lung cancer has not been studied. We determined the prevalence of HFE variants and their impact on cancer phenotypes in lung cancer cell lines, in lung cancer patient specimens, and using The Cancer Genome Atlas (TCGA) database. We found that seven out of ten human lung cancer cell lines carry the H63D or C282Y HFE variant. Analysis of lung cancer specimens from our institute revealed a sex and genotype interaction risk for metastasis in lung adenocarcinoma (LUAD) patients; H63D HFE is associated with less metastasis in males compared to wild type (WT) HFE; however, females with the H63D HFE variant tend to develop more metastatic tumors than WT female patients. In the TCGA LUAD dataset, the H63D HFE variant was associated with poorer survival in females compared to females with WT HFE. The frequency of C282Y HFE is higher in female lung squamous cell carcinoma (LUSC) patients of TCGA than males, however the C282Y HFE variant did not impact the survival of LUSC patients. In the TCGA LUSC dataset, C282Y HFE patients had poorer survival than WT HFE patients. HFE expression level was not affected by HFE genotype status and did not impact patient\u2019s survival, regardless of sex. In summary, these data suggest that there is a sexually dimorphic effect of HFE polymorphisms in the survival and metastatic disease in lung cancer.The homeostatic iron regulator protein HFE is involved in regulation of iron acquisition for cells. The prevalence of two common EGFR) (10\u201335%), K-ras (KRAS) (15\u201325%), and Phosphatase and tensin homolog (PTEN) (4\u20138%). Other gene alterations, e.g. Fibroblast growth factor receptor 1 (FGFR1) amplification (20%) and anaplastic lymphoma kinase (ALK) rearrangement (3\u20137%) are also found in NSCLC [Lung cancer is the second most common cancer in both men and women and is the leading cause of cancer deaths in the US . There ain NSCLC \u20138. InterHFE (homeostatic iron regulator) gene has been interrogated for its relationship to cancer. The HFE gene encodes for a 343-amino acid major histocompatibility complex (MHC) class 1 molecule [HFE gene contains genetic polymorphisms identified as risk factors or disease modifiers for several human diseases such as hereditary hemochromatosis, neurodegenerative diseases, liver disease, and cancers [Cancer cells have a robust iron appetite associated with their higher growth and metabolism . Iron camolecule whose in cancers \u201318.HFE gene occur more frequently in Caucasians than in other races [HFE gene [HFE genotype is around 26% (24% heterozygote and 2.4% homozygote) and 10% (10% heterozygote and 0.44% homozygote), respectively [HFE in the general Caucasian population is 15.3% and 6.8% [HFE variants have been reported in acute lymphoblastic leukemia [HFE variant was observed in malignant gliomas [HFE variant is also a risk factor for colorectal cancer [HFE variant was observed in colorectal cancer [HFE variant is a risk factor for breast cancer [HFE variant in colorectal cancer has been reported as both increased [HFE than male WT HFE GBM patients [Polymorphisms in the er races \u201321. TherHFE gene . One is ectively . The alland 6.8% . Increasleukemia , 23 and leukemia . Increasl cancer , hepatocl cancer \u201330, and l cancer . Increasl cancer and hepal cancer . In addit cancer , 34, 35,t cancer , 35, 36,t cancer , and ovat cancer . The risncreased and decrncreased . In our patients althoughpatients .HFE gene variants, the impact of HFE genotype on lung cancer has not been studied systematically and this knowledge gap is addressed in this study. We interrogated the HFE genotype and/or HFE expression in human lung cancer cell lines, specimens of lung cancer patients, and lung cancer database.Despite the prevalence of lung cancer and the prevalence of the All tested lung cancer cell lines were ordered from American Type Culture Collection or obtained from Dr. Jong K. Yun (Pennsylvania State University College of Medicine). Human lung cancer cell lines were maintained in Roswell Park Memorial Institute (RPMI) media 1640 with 1X Penicillin-Streptomycin and 10% fetal bovine serum (FBS). RPMI media 1640 and other cell culture ingredients were purchased from Life Technologies .HFE genotype of lung cancer cell lines was determined using a restriction enzyme digestion method after PCR. The digested PCR products were run in 5% TBE polyacrylamide gel for HFE genotype as reported [HFE genotyping from plasma of lung cancer patients was performed using same method after genomic DNA purification using DNeasy Tissue kit (Qiagen) according to the manufacturer's instructions. The HFE genotype of select samples was confirmed via DNA sequencing.reported . The HFEHFE genotype as described above. For HFE genotype data analysis, we used only Caucasian lung cancer patients\u2019 samples, because HFE polymorphisms are most prevalent in the Caucasian population. We consented and enrolled 53 adenocarcinoma of the lung and 41 squamous cell lung cancer patients .The de-identified plasma samples and clinical data of human lung cancer patients were obtained from Tumor Bank of Penn State Institute for Personalized Medicine (PSIPM) and approved by Penn State College of Medicine Institutional Review Board (IRB Protocol Number 40532). DNA was purified from the plasma samples by DNeasy Blood & Tissue kit (QIAAGEN) to determine HFE genotype of lung cancer patients was compared with samples from individuals without cancer or with neurological disease in our Institute of Personalized Medicine [HFE gene using Variant Call Format (VCF) file from The International Genome Sample Resource.The Medicine and 1000HFE genotype information was 408. Among 408 samples, there were 307 Caucasian, 36 Black, 6 Asian, and 59 unknown. The total number of TP samples of LUAD patients who had HFE genotype information was 558. Among the 558 samples, there were 381 Caucasian, 52 Black, 8 Asian, 1 American Indian, and 116 unknown. The total number of NB samples of LUSC patients who had HFE genotype information was 322. Among 322 samples, there were 203 Caucasian, 15 Black, 6 Asian, and 98 unknown. The total number of TP samples of LUSC patients who had HFE genotype information was 507. Among 507 samples, there were 300 Caucasian, 28 Black, 9 Asian, and 170 unknown. We accessed HFE gene variant data from Cancer Genomics Hub (CGHub) using GeneTorrent and GTFuse software to extract and download only HFE genes from complete mapped sequence (BAM) files. Genome Analysis Toolkit (GATK) software, based on the GATK best practices pipeline, was used to identify HFE gene variants from the sequences as we previously reported [HFE genotype status and survival analyses using either (i) all subjects, or (ii) all subjects with follow-up times greater than 30 days (both living and deceased). The two approaches produced highly concordant results. Therefore, the results presented here are based on an analysis of all patients.There are two types of lung cancer data in the TCGA database: lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). In the TCGA lung cancer database, we used two samples data from each lung cancer patients. i.e., Blood Derived Normal (NB), Primary Solid Tumor (TP). The total number of NB samples of LUAD patients with corresponding reported \u201344. Durihttps://gdac.broadinstitute.org/). We applied a log transformation to the resulting normalized RSEM (RNA-Seq by Expectation-Maximization) values from RNA-Seq, so gene expression was quantified as log2 (RSEM + 1). Primary tumor samples and matched patient\u2019s normal samples were identified from the TCGA barcodes. Kruskal-Wallis tests or Wilcoxon rank sum tests were used to compare HFE expression values in the following groups: (i) HFE variant vs. HFE wild type (WT), (ii) females vs. males. Separate analyses were performed for primary tumor and matched blood samples in each subject. Additionally, H63D and C282Y variants were considered separately in each tumor type, and the variant status for matched normal samples was determined by the status of the matched primary tumor sample. Kaplan-Meier plots and log rank tests were applied to compare survival times in groups defined by HFE expression level. Survival times were compared for females and males after defining HFE expression groups as high or low based on median expression or based on quartiles. Additionally, multivariable Cox proportional hazards models were fit using either HFE genotype or HFE expression while adjusting for tumor stage (early stage (I/II) vs. advanced stage (III/IV)), age at diagnosis, gender, and smoking (never smoked vs. any smoking history). All survival analyses were performed after restricting to Caucasians only.RNA sequencing (RNA-Seq) and clinical data from the TCGA LUAD and LUSC cohorts were accessed from the Broad Institute\u2019s Firehose GDAC was indicated by Kaplan-Meier survival curve and analyzed by log-rank test. The log-rank test results were further examined using multi-variable Cox proportional hazards regression models, by controlling for several known confounders. R 3.5.0 (R Foundations) was used to conduct all data analyses for HFE expression related study [Comparisons of ed study , and theed study and GrapHFE variant and 1 had the C282Y HFE variant (Table 1). Only 1 large cell lung cancer cell line did not have a HFE gene variant and the only squamous cell carcinoma cell line was heterozygote for C282Y.Of the ten lung cancer cell lines investigated, 6 expressed the H63D HFE gene variants in lung cancer patients seen in our Penn State Hershey Medical Center (PSHMC), and publically available lung cancer databases. The characteristics of lung cancer patients, such as median age, age range, and male: female ratio, are summarized in Table 2.We determined the frequency of HFE polymorphisms compared to other races [HFE gene variant was 32.0% and 13.2%, respectively . The allele frequency of H63D HFE was 20.8% and 8.5% for C282Y HFE in lung adenocarcinoma, and 7.3% for H63D HFE and 7.3% for C282Y HFE in lung squamous cell carcinoma. Statistical analysis revealed a lower frequency of H63D HFE alleles in squamous cell carcinoma compared to adenocarcinoma (p = 0.007). In comparison to non-cancer population adenocarcinoma tended to have higher C282Y HFE allele frequency (p = 0.08) compared to 1000Genome database. Squamous cell carcinoma had significantly lower H63D HFE allelic frequency than the PSHMC control samples (p = 0.0336) or 1000Genome data (p = 0.04).We limited our analysis to Caucasians because of the higher frequency of er races \u201321. AmonHFE genotype or HFE alleles (S2 Table). The summary of allele frequencies for H63D HFE and C282Y HFE in lung cancers and controls are summarized in Fig 1.There were no differences between TCGA lung cancer samples and non-cancer population for the frequency of HFE genotype data were stratified by sex. There was an increase in the C282Y HFE allelic frequency in female LUAD patients in the TCGA database (p = 0.0474) (Table 3) but not in the PSHMC database (S3 Table). There were no sex differences in the H63D HFE allele groups.Because distinct male: female ratios exist in two lung cancer types , the HFE genotype and HFE mRNA expression in TCGA lung cancer data. There were no differences between WT and H63D or C282Y HFE variants in matched normal or primary tumors of LUAD and LUSC lung cancer patients in two sample t-test . The level of HFE expression was not different between males and females in matched normal or primary tumors of LUAD and LUSC lung cancer patients in two sample t-test . There were no HFE expression differences between WT and HFE variants in male or female LUAD and LUSC in the TCGA lung cancer dataset .We evaluated the association between HFE polymorphisms but when we stratified for sex, there were significant differences between WT and HFE polymorphisms (Table 4). None of the H63D HFE male adenocarcinoma patients developed metastases while 50% of WT HFE male patients did (p = 0.0189); whereas in females the findings were opposite with H63D HFE carriers tending to develop more metastases than WT HFE (p = 0.0596). With the C282Y HFE variant, male adenocarcinoma patients tend to develop more metastases compared to WT HFE male carriers (p = 0.0672) while there is no difference in females. In squamous cell lung cancer, neither of the HFE variants (H63D and C282Y) or sex impacted metastatic rate.Metastatic disease was only available in the PSHMC database. The incidence of metastasis in lung cancer patients at PSHMC revealed no statistical difference between WT and HFE , or WT and C282Y HFE lung cancer patients who enrolled at PSHMC .There was no survival difference between WT and H63D HFE variants compared to WT HFE patients (p = 0.0763) . When the analysis focused on H63D homozygotes compared to WT HFE patients, the survival difference was marginally significant (p = 0.0539) . The survival for WT HFE patients and C282Y HFE patients was not different . The Kaplan-Meier survival curve for LUSC patients revealed a significant survival difference between WT HFE patients and C282Y HFE patients (p = 0.0067), but not between WT HFE and H63D HFE variants .In TCGA database, the Kaplan-Meier survival curve for lung adenocarcinoma patients showed a trend toward poorer survival in H63D HFE gene variant and overall survival by further controlling other possible factors (tumor stage etc.) that could contribute to the overall survival. The effect of C282Y HFE variant on LUSC patient survival remains significant after controlling for age, tumor stage, and smoking status in multiple Cox regression (Table 5).We performed multiplex Cox regression to double check the bivariate associations (Kaplan-Meier curve) between HFE genotype we found that male LUAD patients with H63D HFE had no survival difference compared to WT HFE and female LUAD patients with H63D HFE had significantly poorer survival compared to WT HFE (p = 0.0455) . The dramatic difference in survival for male LUSC patients with C282Y HFE compared to WT HFE did not reach statistical significance (p = 0.0665); however, female LUSC patients with C282Y HFE had significantly poorer survival compared to WT HFE (p = 0.0283) .When we stratify the survival data by sex and HFE expression and sex. The survival pattern was not different regardless of HFE expression levels in lung cancer patients. There were no survival differences between lower 50% and upper 50%, or between lower 25% and upper 75% in TCGA lung cancer dataset . In general, the survival curve between males and females was not different regardless of HFE expression levels in lung cancer patients . However, when HFE expression level is in the upper 50%, male LUAD and LUSC patients tend to had poorer survival than females .We further compared the survival of lung cancer patients based on HFE polymorphisms in lung cancer cell lines and human lung cancer patients. We found 7 of 10 lung cancer cell lines had the H63D or C282Y variant of the HFE gene. This result provided motivation to interrogate lung cancer samples from our institute and existing database such as TCGA for the frequency and impact of HFE gene variants on lung cancer.The present study determined the frequency of HFE genotype and sex differences for survival and metastatic disease. In the TCGA lung cancer database, adenocarcinoma patients with H63D HFE trended to have poorer survival outcomes than WT HFE, but when the outcome data were stratified by sex the difference in survival outcome became statistically significant for females. The TCGA database does not include information on metastatic disease so we used our smaller internal database (PSHMC) and found a sex and genotype dependent effect for adenocarcinoma patients; males with the H63D HFE variant had less metastasis and females with the H63D HFE variant had more metastasis. There is no frequency increase in H63D HFE in the squamous cell cancer population or impact on the disease noted in our studies. These interesting results suggest that H63D HFE variant protects lung cancer metastasis in males, but enables metastasis in females. As far as we know, this is the first report for the impact of HFE variants and sex on lung cancer patients\u2019 outcome.There are significant HFE gene, the C282Y HFE variant is also found in the general population although at a lower prevalence. This gene variant has received considerable attention in hepatic cancers because of its tendency to be associated with hemochromatosis, the iron overload disease. In the TCGA database, individuals with squamous cell carcinoma that carry C282Y HFE have poorer survival than WT HFE. Moreover, when stratified for sex, it appears the females with C282Y are driving the significant difference as their survival outcome is much worse than males. There is no impact of this genotype of metastatic disease in the squamous cell cancer population. There is no statistically significant effect of C282Y HFE on the adenocarcinoma population, but there is a trend toward greater frequency of metastatic disease in males compared to females although the sample size is small. All these data suggest that females with C282Y HFE are a risk factor for lung squamous cell carcinoma. In our previous PSHMC GBM samples study, we found poorer survival in female metastatic brain tumor patients with C282Y HFE than WT HFE patients or male metastatic brain tumor patients with C282Y HFE [HFE is both cancer type and sex dependent.In addition to the H63D variant of the 282Y HFE . The datHFE genotype and sex effect for lung cancer patient\u2019s survival and metastatic disease, this is not related with the frequency of HFE genotype/alleles. For example, we observed higher frequency of C282Y HFE allele in female lung adenocarcinoma patients than male patients in TCGA; however, there was no survival difference between female C282Y HFE and male C282Y HFE lung adenocarcinoma patients. Instead, we found survival difference between female H63D HFE adenocarcinoma patients and female WT HFE adenocarcinoma patients.Although we found HFE gene expression was not different between WT HFE and HFE variants of lung cancer patients in TCGA lung cancer data. In addition, HFE gene expression level did not impact survival of TCGA lung cancer patients even when stratified for sex. These results suggest that poor survival in female lung adenocarcinoma with H63D HFE and female lung squamous cell cancer with C282Y HFE is not due to the expression level of HFE variants or frequency of HFE genotype but rather the function of HFE variant itself. There are many reports for the function of H63D or C282Y HFE in human cancer. For example, at the cellular level, the HFE H63D variant alters cholesterol metabolism [HFE variants in LUAD and LUSC in animal models as a future direction.In the present study, the level of tabolism , Endoplatabolism and altetabolism , 49 incltabolism . Mouse Htabolism . The C28HFE genotype and cancer patient survival. When we compare the key variables between the TCGA lung cancer and the PSHMC samples, there are difference in age at diagnosis , smoking status , and the vital status suggesting the PSHMC and TCGA cohorts have important distinctions. Therefore, we only showed Kaplan-Meier survival curve of PSHMC samples without Cox-regression results.There are no consistent results between PSHMC and TCGA lung cancer samples for HFE gene variant is associated with poor survival and increased metastasis in female, but not male, LUAD patients. Although the frequency of C282Y HFE gene variant is higher in female LUAD patients than males, there was no gene effect on survival in these patients. However, the C282Y HFE gene variant is associated with poor survival in LUSC patients, especially females. The present findings indicate that there is a distinct impact of H63D and C282Y HFE variants in two different subtype of human lung cancers and impact of the genotype is influenced by sex.In summary, the H63D S1 FigHFE gene expression based on WT HFE vs. H63D HFE in matched normal or primary tumor LUAD patients. (B) HFE gene expression based on WT HFE vs. C282Y HFE in matched normal or primary tumor LUAD patients. (C) HFE gene expression based on males vs. females in matched normal or primary tumor LUAD patients. P value was calculated from Wilcoxon rank sum tests to compare HFE expression values in the HFE mutant vs. HFE wild type.(A) (PPTX)Click here for additional data file.S2 FigHFE gene expression based on WT HFE vs. H63D HFE in matched normal or primary tumor LUSC patients. (B) HFE gene expression based on WT HFE vs. C282Y HFE in matched normal or primary tumor LUSC patients. (C) HFE gene expression based on males vs. females in matched normal or primary tumor LUSC patients. P value was calculated from Wilcoxon rank sum tests to compare HFE expression values in the HFE mutant vs. HFE wild type.(A) (PPTX)Click here for additional data file.S3 FigHFE or H63D HFE. (B) Survival curve of LUAD patients with WT HFE or C282Y HFE. (C) Survival curve of LUSC patients with WT HFE or H63D HFE. (D) Survival curve of LUSC patients with WT HFE or C282Y HFE. Statistical analysis was performed by log-rank test and indicated as p value. Censored record is indicated as + in the graph.(A) Survival curve of LUAD patients with WT (PPTX)Click here for additional data file.S4 FigHFE or heterozygote or homozygote H63D HFE. (B) Survival curve of LUSC patients with WT HFE or heterozygote or homozygote C282Y HFE. LUSC with C282Y HFE heterozygote had poorer survival than WT HFE (p = 0.0052). Statistical analysis was performed by log-rank test and indicated as p value. Censored record is indicated as+ in the graph.(A) Survival curve of LUAD patients with WT (PPTX)Click here for additional data file.S5 FigHFE gene expression. (B) Survival curve of TCGA LUAD patients between lower 25% and upper 75% of HFE gene expression. (C) Survival curve of TCGA LUSC patients based on lower 50% or upper 50% of HFE gene expression. (D) Survival curve of TCGA LUSC patients between lower 25% and upper 75% of HFE gene expression. P value was calculated from log rank tests to compare survival times in groups defined by HFE expression level.(A) Kaplan-Meier survival curve of TCGA LUAD patients based on lower 50% or upper 50% of (PPTX)Click here for additional data file.S6 FigHFE gene expression at lower 25% (A) or lower 50% (B) or upper 50% (C) or upper 75% (D). Log rank tests were used to compare survival times in groups defined by HFE expression level.Survival curve between males and females of TCGA LUAD patients based on (PPTX)Click here for additional data file.S7 FigHFE gene expression at lower 25% (A) or lower 50% (B) or upper 50% (C) or upper 75% (D). Log rank tests were used to compare survival times in groups defined by HFE expression level.Survival curve between males and females of TCGA LUSC patients based on (PPTX)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file."} +{"text": "High grade serous ovarian carcinoma (HGSOC) is the most common subtype of epithelial ovarian cancers (EOC) with poor prognosis. In most cases EOC is widely disseminated at the time of diagnosis. Despite the optimal cytoreductive surgery and chemotherapy most patients develop chemoresistance, and the 5-year overall survival being only 25\u201335%.Here we analyzed the gene expression profiles of 10 primary HGSOC tumors and 10 related omental metastases using RNA sequencing and identified 100 differentially expressed genes.The differentially expressed genes were associated with decreased embryogenesis and vasculogenesis and increased cellular proliferation and organismal death. Top upstream regulators responsible for this gene signature were NR5A1, GATA4, FOXL2, TP53 and BMP7. A subset of these genes were highly expressed in the ovarian cancer among the cancer transcriptomes of The Cancer Genome Atlas. Importantly, the metastatic gene signature was suggestive of poor survival in TCGA data based on gene enrichment analysis.By comparing the gene expression profiles of primary HGSOC tumors and their matched metastasis, we provide evidence that a signature of 100 genes is able to separate these two sample types and potentially predict patient survival. Our study identifies functional categories of genes and transcription factors that could play important roles in promoting metastases and serve as markers for cancer prognosis. Ovarian cancer is the seventh most common cancer in females worldwide, and the fifth most common in Europe . In EuroDespite large number of studies profiling the transcriptome of EOC primary ovarian tumors, only limited number of reports have compared gene expression between primary tumors and their matched metastases. These studies have identified differentially expressed genes implicated in oncogenesis, metastasis, p53 signaling , cell adAim of this study was to study the differences in the gene expression profiles of histologically validated HGSOC metastases compared to primary tumors using RNA-Seq. Samples were collected during the same cytoreductive surgery before chemotherapy. To validate our results, our data was compared to TCGA database and to the known four molecular subtypes of HGSOC described by Tothill et al. and TCGA , 11.n\u2009=\u20094) or IV (n\u2009=\u20096). Histologically all tumors were high grade serous ovarian carcinomas. Samples were frozen in liquid nitrogen and stored at \u2212\u200980\u2009\u00b0C until RNA preparation. For qRT-PCR analyses paired primary tumors and omental samples of six additional HGSOC patients were included. Those patients\u2019 ages ranged from 46 to 86 (median 67\u2009years) and FIGO stages of the patients were IIIC (n\u2009=\u20094) or IV (n\u2009=\u20092). The samples for qRT-PCR were also collected in the same cytoreductive surgery before chemotherapy like samples for RNA\u2013seq.Samples of primary adnexal tumor and paired omental metastases of 10 HGSOC patients were included in the study. Primary and metastatic samples were collected in the same cytoreductive surgery before chemotherapy in each patient in Kuopio University Hospital between 2004 and 2013. The patients\u2019 ages ranged from 44 to 75 (the median 58\u2009years). All patients were FIGO stage IIIC followed by DNase treatment using the Turbo DNase kit (Thermo Scientific). Ribosomal RNA was depleted using the Ribo-Zero Gold (Illumina). Libraries were prepared as previously described by Kaikkonen et al. . BrieflyAMHR2 (qHsaCEP0041252), GATA4 (qHsaCIP0028312), MAL (qHsaCEP0039522), MYOCD (qHsaCEP0058240), NR5A1 (qHsaCIP0028304), PPIA (qHsaCEP0041342), PROK1 (qHsaCEP0024916), SFRP2 (qHsaCEP0052530), WIPF3 (qHsaCEP0051213), WNT5A (qHsaCIP0028356). Relative expressions were quantified with 2-\u0394\u0394CT method [PPIA as the reference gene.RNA was isolated using TRI-reagent (Thermo Scientific). One microgram of RNA was treated with DNAse I (Thermo Scientific) and converted into cDNA using RevertAid reverse transcriptase (Thermo Scientific) and random hexamers (Thermo Scientific). Analysis of mRNA levels were done using StepOnePlus Real-Time PCR System (Life technologies), TaqMan Universal PCR Mastermix (Applied Biosystems) and gene-specific Prime PCR Probe Assays (BioRad): www.qiagen.com/ingenuity) was used. For IPA\u00ae upstream regulator analysis, the top transcriptional regulators and growth factors were chosen based on most significant P-values (P\u2009<\u20093.5E-04) and a clear predicted activation state (\u2212\u20092\u2009<\u2009activation z-score\u2009>\u20092).RNA-Seq was mapped using tophat allowing up to two mismatches and reporting only one alignment for each read. Poor quality reads were filtered out and tag per base value was set to 3. RefSeq expression was quantified using \u2018analyzeRNA.pl\u2019 program in HOMER . DiffereThe experiments performed in this study are available in GEO under the accession number GSE98281.Survival time and status and RSEM RNA-seq data for each TCGA OV sample was obtained from firehose GDAC, doi:10.7908/C11G0KM9.P-value was computed using 1000 random permutations of genes. Same amount of genes as the observed gene set was used. The observed pathway score was compared with the random permutations of a gene set size and empirical P-value computed as the number of higher/lower scores in the permuted set divided by the total number of permutations. Upregulated and downregulated metastasis genes were split to individual gene sets to account for directionality of gene set enrichment. Enriched samples were required to have significant enrichment of both gene sets with P-value <\u20090.001 for Kapplan Meier survival analysis.One hundred differentially expressed genes between primary tumors and metastases, defined as metastatic signature were used in the analysis. The gene set variation analysis (GSVA) , availabP-values.The R package \u2018survival\u2019 was used to draw Univariate Kaplan Meier curves comparing samples with significant enrichment of metastatic signature to rest of the samples, as indicated above. The log-rank test was computed for significance evaluation between the groups. Univariate cox proportional hazard analysis was performed for TCGA data for each 100 metastatic genes and BH method was used to adjust AMHR2), GATA binding protein 4 (GATA4), myelin and lymphocyte protein (MAL), myocardin (MYOCD), nuclear receptor subfamily 5 group A member 1 (NR5A1), prokineticin 1 (PROK1), secreted frizzled related protein 2 (SFRP2), WAS/WASL interacting protein family member 3 (WIPF3) and Wnt family member 5A (WNT5A) using qPCR from 6\u2009+\u20096 samples of them exhibiting downregulation in the omental samples , 4/6 GATA4-targets , 3/4 NR5A1-targets were found to contain the respective transcription factor motif within the gene promoter was found associated with a significant negative z-score (thus likely to be repressed) and bone morphogenetic factor 7 (BMP7) with a positive z-score contains publically available data about the genetic alterations of different cancers and also linkage to clinical features and prognosis. TCGA database contains information on the key genomic changes in over 30 different cancer types and also collection of primary ovarian tumors at the initial site of cancer, which allows comparison between different cancer types based on their gene expression profile. To see which of our differentially regulated genes were highly expressed in ovarian tumors, we compared the expression level of the 100 genes identified in the study throughout the TCGA cancer types. The analysis revealed that many of the embryonic and cell development genes are fairly high expressed in ovarian cancer including P-value <\u20090.05), including AMHR2, GATA4, MAL, SFRP2, Family With Sequence Similarity 19 Member A2 (FAM19A2), Paired Box\u00a05 (PAX5) and Proprotein Convertase Subtilisin/Kexin Type 6 (PCSK6) where they conducted a whole tumor gene expression profiling of 285 predominantly high-grade and advanced-stage serous cancers of the ovary, peritoneum and fallopian tubes . The autN8) Fig. b. AltogeNR1H4, CADPS, STAR, SFRP2 and EPYC were also observed to be differentially expressed in the similar direction in the earlier studies [FOXL2, GATA4, NR5A1, AMHR2, MAL and WIPF3. Of these, the first three were further identified as potential upstream regulators that could explain the observed gene expression patterns. Accordingly, the GATA4 has been shown regulate genes involved in embryogenesis and development of the female reproductive organs, testes, GI-tract, heart and lungs [GATA4 [GATA4 is downregulated in our metastatic gene signature in HGSOC. Statistically significant higher methylation leading to the loss of GATA4 expression in endometrioid type compared to serous ovarian adenocarcinoma has been reported [GATA4 expression and patient age, histologic type, histologic grade, stage of the disease or survival in ovarian surface epithelial carcinomas has been reported [NR5A1 transcription factor, was also downregulated in omental samples. It encodes a human steroidogenic factor 1-protein (hSF1) that is involved in gonad development in both males and females [NR5A1 are associated with primary ovarian insufficiency [FOXA2, that has demonstrated favorable prognosis based on TCGA data [DLK1, GATA4, MAFA, MYOCD, NR1H4 and WNT5). However, FOXA2 was not differentially expressed in our data but rather another member of the FOX-family that encodes for transcription factor that is involved in all stages of ovarian development and function, FOXL2 [FOXL2-positive cells were found mainly in primary and secondary ovarian tumors and very few in peritoneal seeding sites suggesting that local tissue environment could be responsible for its omental downregulation [This is the first study to compare gene expression between primary EOC tumors and their matching omental metastases using RNA-seq, allowing more sensitive and deeper characterization of transcriptome compared to microarray . In line studies , 8. In cnd lungs . Loss ofnd lungs , clear cnd lungs , 26 and nd lungs ovarian s [GATA4 . This isreported . Howeverreported . Another females . hSF1 ex females and mutaficiency . The thiCGA data and was n, FOXL2 . Whethern, FOXL2 . In a regulation . On the gulation . Future The identification of different ovarian cancer subgroups could allow for more personalized treatments and is therefore heavily investigated. Previous molecular subtyping systems defined by TCGA and Tothill studies , 11 haveMAL and FAM19A2 in our analysis. However, among the five other genes with prognostic value, genes associated with better prognosis were downregulated and genes with poorer prognosis were upregulated (PAX5 and SFRP2) in the metastatic samples. This could reflect subtype differences of the EOCs, as patients in our study were limited to HGSOCs only. Recent reports have also identified markers related to recurrence in ovarian cancer primary tumors. These further identified networks related to TP53 and TGF-\u03b2 signaling, cell cycle, leukocyte migration and cellular adhesion [Finally, survival analysis based on gene set enrichment analysis of TCGA primary tumors expression profiles revealed that the differentially regulated genes identified in this study could be indicative of poorer survival. This is in line with previous report based on 19 matched primary and omental metastatic tumors from 3 different serous adenocarcinoma types . In contadhesion , 42. EviIn this study we provide evidence that the gene expression profile of primary HGSOC tumors differs from their matched metastases, and that the 100 differentially expressed genes identified could nominally predict patient survival. Identified functional categories of genes and transcription factors could play important roles in promoting metastases and serve as markers for cancer prognosis. These findings serve candidates for future research and could lead to improved treatments for HGSOC in the future.Additional file 1: Figure S1. Comparison of the expression of 9 differentially regulated genes from additional 6 patients by qPCR (white bars) was in line with the RNA-Seq results (black bars).Additional file 1: Table S1. The differentially expressed genes identified in our analysis.Additional file 2: Table S2. Gene Ontology Analysis (Diseases or Functions Annotation) of differentially expressed genes performed using Ingenuity Pathway Analysis.Additional file 3: Table S3. Number of upstream transcription factor motifs predicted within (+/\u2212\u20091.5\u2009kb of TSS) or around (+/\u2212\u200950\u2009kb from TSS) promoters of IPA predicted target genes.Additional file 4: Table S4. Upstream regulators identified in our data using Ingenuity Pathway Analysis.Additional file 5: Table S5. Univariate cox proportional hazard analysis for our 100 metastatic genes performed from TCGA data."} +{"text": "SRL1, ROC5, OsRRK1, SLL2, CLD1, OsZHD1/2, and NRL1, structure and processes of sclerenchyma cells by SLL1 and SRL2, leaf polarity by ADL1, RFS and cuticle formation by CFL1, and CLD1. Many of above mentioned and several other genes interact in a complex manner in order to sustain cellular integrity and homeostasis for optimum leaf rolling. While, leaf size is synchronized by multifarious interaction of PLA1, PLA2, OsGASR1, and OsEXPA8 in cell division, NAL1, NAL9, NRL1, NRL2 in regulation of number of veins, OsCOW1, OsPIN1, OsARF19, OsOFP2, D1 and GID in regulation of phytohormones and HDT702 in epigenetic aspects. In this review, we curtailed recent advances engrossing regulation and functions of those genes that directly or indirectly can distress leaf rolling or size by encoding different types of proteins and genic expression. Moreover, this effort could be used further to develop comprehensive learning and directing our molecular breeding of rice.Yield is majorly affected by photosynthetic efficiency. Leaves are essential structure for photosynthesis and their morphology especially size and shape in a plant canopy can affect the rate of transpiration, carbon fixation and photosynthesis. Leaf rolling and size are considered key agronomic traits in plant architecture that can subsidize yield parameters. In last era, a number of genes controlling leaf morphology have been molecularly characterized. Despite of several findings, our understanding toward molecular mechanism of leaf rolling and size are under-developed. Here, we proposed a model to apprehend the physiological basis of different genes organized in a complex fashion and govern the final phenotype of leaf morphology. According to this leaf rolling is mainly controlled by regulation of bulliform cells by Rice is a model plant of monocots and an important crop that feeds more than half of the population around the globe. According to estimates, food supply will be insufficient as growth rate of population and increase in yield is not harmonious . Yield iThe leaf is a major photosynthetic organ in plants and its morphology such as size and rolling are key components in plant architecture that significantly affect crop yield . Leaf roThe ideotype breeding of \u2018Super rice\u2019 suggests that uppermost three leaves should be erect, long, rolled (V-shaped), narrow, and dark green in color . EngineeAdvancements in molecular methodologies lead to discovery of several leaf rolling genes that have been cloned and functionally characterized in rice. We have categorized those genes based on their part, which they play in final appearance of leaf.ADL1 (ADAXIALIZED LEAF 1) gene was isolated from adl1 mutant using positional cloning, and it encodes a CALPAIN-LIKE CYSTEINE PROTEINASE. Deficiency of CALPAIN-LIKE CYSTEINE PROTEINASE leads to rolling of leaves abaxially. Morphological analysis revealed bulliform cells specifically appear only on the adaxial side of leaf normally were also found on the abaxial side in adl1 mutant (SLL1 (SHALLOT-LIKE 1) and overexpression of OsAGO7 (ARGONATUE) caused leaf rolling (ROLLED FINE STRIPED (RFS) also showed leaf rolling phenotypes in rice due to poor development of vascular bundles on adaxial side of leaf. Knock down of key elements of CHD3/Mi-2 (chromatin remodeling factor of RFS) caused severe leaf rolling in rfs-1 mutant encodes a putative GPI (GLYCOSYLPHOSPHATIDYLINOSITOL) anchored protein in rice that is located in plasma membrane. The loss-of-function mutant of SRL1 exhibits rolling of leaves adaxially due to an augmented number of bulliform cells on adaxial surface of leaf. Further studies demonstrated SRL1 negatively normalizes the expression of genes encoding vacuolar H+-PYROPHOSPHATASE and H+-ATPASE that usually impede the development of bulliform cells (OsHox32 that belongs to HD-ZIPIII (HOMEODOMAIN LEUCINE ZIPPER) gene family had a similar phenotype to that of loss-of-function mutant of SRL1. However, besides increasing, reduced number of bulliform cells are also responsible for leaf rolling mutant displayed defects of leaf rolling and is allelic to SRL1. CLD1/SRL1 encodes a GPI anchored protein that plays its role in formation of cell wall. The loss-of-function mutant of CLD1/SRL1 showed lesser contents of lignin and cellulose in epidermis of bulliform cells. Defects in cell wall formation cause more rapid water loss and reduce water retaining capacity of leaves or its homolog OsZHD2 and a defect in LC2 (LEAF INCLINATION 2) induced more number of bulliform cells that causes abaxial rolling of leaves mutant in rice exhibited adaxial curling of leaves phenotype due to underdeveloped size of bulliform cells. Mutation in hal1 mutant also affected size of leaf blade and spikelet (RL14 (ROLLING LEAF 14) encodes a 2OG-Fe (OXYGENASE PROTEIN) that is convoluted in formation of secondary cell wall of leaf (NRL1 (NARROW AND ROLLED LEAF 1) , encodes LEAF 1) . OsMYB10ynthesis . TransgeROC5 (RICE OUTER CELL SPECIFIC 5) and its downstream gene (PFL) PROTODERMAL FACTOR LIKE (ACL1 (ABAXIALLY CURLED LEAF 1), its homologous gene ACL2 (ROLLED AND ERECT LEAF 1 (REL1) (REL2 ((LATERAL ORGAN BOUNDARIES DOMAIN 3-7) OsLBD 3-7 ((NARROW LEAF 7) NAL7 (NAL2/3 ((BRASSINOSTEROID INSENSITIVE 1-ASSOCIATED KINASE 1) OsI-BAK1 ((AUXIN RESPONSE FACTOR) OsARF18 mutant revealed positive correlation of bulliform cell area and LRI (leaf rolling index) in rice showed more adaxial curling of leaves in order to increase the photosynthetic activity through regulation of bulliform cells and other related genes (OsRRK1 (RECEPTOR-LIKE CYTOPLASMIC KINASE 1) also showed an erect morphology with decreased number and size of bulliform cells in order to get high seed setting , REL2 (Y1) NAL7 , NAL2/3 in cuticle development exhibited its involvement in leaf rolling. CFL1 encodes a WW DOMAIN PROTEIN that was isolated from cfl1 mutant with curling leaf phenotype. Moreover, overexpression of OsMYB103L encodes a transcription factor R2R3MYB that regulates contents of cellulose and mechanical strength of leaves is major target of miRNA166 and play role in development of xylem and transpiration rate. Knockdown of miRNA166 in short tandem target mimic (sttm166) line resulted in rolling of leaf, which had smaller bulliform cells and reduced stomatal conductance (DICER-LIKE 1 (DCL1) is required for the miRNA biogenesis and has pivotal influences on plant growth and development. Down-regulation of OsDCL1 in RNAi lines leads to a phenotype of narrow leaf blade encode WUSCHEL-RELATED HOMEOBOX PROTEIN in rice that are duplicate orthologs of genes NS1 (NARROW SHEATH 1) and NS2 in maize, respectively. The nal2:nal3 double mutant leaves contains decreased number of veins with extremely narrow leaves in rice and candidate gene for was NAL1 . Yield pmosome 7 . These eNAL1, NRL1, and AVB (ABNORMAL VASCULAR BUNDLES) all are reported to be involved in cell division, implying these genes probably affect cell division to regulate leaf size homeostasis to sustain osmoregulation, and loss-of- function of OsCCC1 leads to narrow leaves illustrating cell division also accounts for narrow leaves phenotype (PLA1 (PLASTOCHRON 1) and PLA2 encode cytochrome P450 and an RNA-binding protein, respectively, and both of them were reported to regulate leaf size mainly due to increase in cell size that belongs to family of GAST [GIBBERELLIN (GA)-STIMULATED TRANSCRIPT] showed higher expression in regions of cell proliferation and increase leaf blade size due to increase in cell length. Its mRNA expression could be triggered by an exogenous application of gibberellins (OsGIF1 (GRF-INTERACTING FACTOR 1) influenced grain production and size of leaves in rice by regulating leaf cell size and ANGUSTIFOLIA3 (AN3) in rice and Arabidopsis, respectively. A loss-of-function mutant MKB3 exhibited narrowed- and rolled-leaf phenotype (OsEXPA8 (EXPANSIN 8) produced improved root system, enhanced leaf number and enlarged leaf size in rice. Further analysis of OsEXPA8 line showed increased lignin content in cell wall and enhanced length of leaf cells (OsEBS (ENHANCING BIOMASS AND SPIKELET NUMBER) in rice caused increase in plant height, leaf size and spikelet number per panicle due to increase in cell number (dissociator (Ds) was inserted into gene OsCYP96B4 (CYTOCHROME P450 96B4). It showed defects in plant height and length of leaf sheath cells gene is identical to NAL7 that encodes a FLAVIN-CONTAINING MONO-OXYGENASE protein and indicated resemblance with YUCCA in Arabidopsis and FLOOZY in petunia, which encode for auxin biosynthesis showed that it can serve as an auxin efflux facilitator and FIB (FISH BONE) both were involved in auxin biosynthesis and mutants with reduced expression of TDD1 or FIB showed narrow leaves phenotype OsOFP2 (OsGA2ox6 ((DWARF 1) D1 ((GIBBERELLIN-INSENSITIVE DWARF 2) GID2 OsOFP2 , OsGA2oxOsGA2ox6 , (DWARF RF 1) D1 , and (GI 2) GID2 all wereNAL1 regulates both plant height and leaf size. DNL1 (DWARF AND NARROW LEAF 1) allelic to NRL1, is a QTL for leaf size and plant height (SLL1 gene displayed narrow rolled leaves and reduced plant height (NAL2/3. Overexpression of NAL2/3 leads to dwarf phenotype in rice (dwarf 1 (d1) mutant have wider leaves phenotype and overexpression of SG1 (SHORT GRAIN 1) results in reduced plant height and increased leaf size gene family that plays an important function in histone modifications and plant gene expression. Down-regulation of HDT702 results in narrow leaves, indicating that histone modifications are also involved in regulation of leaf size (Histone modifications are important part of epigenetic mechanisms and have determinant role in controlling leaf size. Different stresses act as stimulus and change the genic expression level by various epigenetic mechanisms, e.g., DNA methylation, histone modifications and miRNA . HDT702 eaf size .In past several years, molecular and genomic studies disclosed important advancements in identification of genes or QTLs controlling leaf morphology Table . InnovatBiotic and a-biotic stresses such as high temperature, drought, fungus and insects cause severe yield losses to plants . Plant mPX and AA wrote the review. XW read and approved the contents. BH helped in literature and reference digestion.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Cancer cells often accumulate spontaneous and treatment-induced DNA damage i.e. potentially lethal DNA double strand breaks (DSBs). Targeting DSB repair mechanisms with specific inhibitors could potentially sensitize cancer cells to the toxic effect of DSBs. Current treatment for glioblastoma includes tumor resection followed by radiotherapy and/or temozolomide (TMZ) \u2013 an alkylating agent inducing DNA damage. We hypothesize that combination of PARP inhibitor (PARPi) with TMZ in glioblastoma cells displaying downregulation of DSB repair genes could trigger synthetic lethality. In our study, we observed that PARP inhibitor (BMN673) was able to specifically sensitize DNA ligase 4 (LIG4)-deprived glioblastoma cells to TMZ while normal astrocytes were not affected. LIG4 downregulation resulting in low effectiveness of DNA-PK\u2013mediated non-homologous end-joining (D-NHEJ), which in combination with BMN673 and TMZ resulted in accumulation of lethal DSBs and specific eradication of glioblastoma cells. Restoration of the LIG4 expression caused loss of sensitivity to BMN673+TMZ. In conclusion, PARP inhibitor combined with DNA damage inducing agents can be utilized in patients with glioblastoma displaying defects in D-NHEJ. Glioblastoma is the mOne of the hallmarks of the cancerous cells is genomic instability responsible for accumulation of further genome rearrangements and tumor progression . DevelopDNA double-strand breaks (DSBs) are the most toxic among DNA lesions and can be responsible for genome rearrangements leading to genomic instability, neoplastic transformation and cell death . DSBs caIn order to utilize personalized synthetic lethality approach we determined the expression profile for 3 patient-derived glioblastoma primary cell lines and compared it to the profile of normal human astrocytes (NHA). The subject of our interest were 15 genes involved in DSB repair pathways . Significant changes in the mRNA expression profile of LIG4 was found between glioblastoma cell lines and NHA Figure . DecreasLoss of heterozygosity (LOH) in chromosome loci 10q, 10p and 22q were reported to be one of the most commonly occurring abnormalities during astrocytoma progression \u201320. Ther+, PI-) and then shifting to Q2 quadrant what would characterize slow externalization of phosphatidylserine and prolonged annexin V binding which is typical for apoptosis used either alone or in combination with alkylating agent (TMZ), double staining with propidium iodide (PI) and annexin V was used. Annexin V staining in conjunction with vital dye (PI) distinguishes viable cells from dead, and also early apoptotic cells from necrotic cells. When compared to individual agents, the combination of BMN673 + TMZ exerted significantly stronger anti H6 and H7 glioblastomas effect with only minimal toxicity to normal astrocytes Figure . The flos Figure can be used as a marker of DSBs . TMZ treNeutral comet assay was also employed to detect DSBs after treatment with BMN673 and/or TMZ. After treatment with individual drugs only TMZ enhanced the percentage of DNA in tails of H7 cells in comparison to NHA cells Figure . CombinaTo determine the role of reduced expression of LIG4 in sensitivity of glioblastoma cells to BMN673+TMZ, H7 cells were transfected with the plasmid carrying LIG4 cDNA followed by treatment with the drugs. Elevated expression of LIG4 resulted in resistance of H7 glioblastoma primary cell line to BMN673 + TMZ was detected only in approximately 4% of glioblastomas in The Cancer Genome Atlas (TCGA) database inhibitiIn summary, this study implicates potential therapeutic effect of PARPi used in combination with DNA-damaging agents in D-NHEJ-deprived glioblastoma cells. Therefore patient pre-selection based on expression of DNA repair genes may be applied for personalized medicine approach to improve the effectiveness of anti-glioblastoma therapy.2 at 37\u00b0C. The normal human astrocytes NHA (Lonza) were grown in ABM Basal Medium supplemented with AGM BulletKit (Lonza) and cultured according to the protocol provided by manufacturer.Glioblastoma specimens, histopathologically classified as clinical stage IV, were obtained from patients of Department of Neurosurgery, Surgery of Spine and Peripheral Nerves, Medical University of Lodz and Department of Neurosurgery, Medical University of Lodz . Cell cultures derived from specimens were established in the Laboratory of Molecular Genetics, University of Lodz and named H3, H6 and H7. After several washes tissue fragments were minced with scalpel and cells were filtered through 70 \u03bcM pore size cell strainer. Glioblastoma cells were cultured in DMEM medium supplemented with 10% FBS (Lonza), 100 IU/ml penicillin, 100 \u03bcg/ml streptomycin (Lonza) and gentamycin 50 \u03bcg/ml (Lonza) in a humidified atmosphere containing 5% COLoss of heterozygosity (LOH) analysis was performed using microsatellite markers. The aim of the experiment was to verify presence of genetic aberrations specific for glioblastoma cells in suspected cancerous tissue. The samples were examined for LOH using sets of DNA samples isolated from tumor bulk specimen and corresponding cell culture and peripheral blood samples (PBMC). Isolation and purification of genomic DNA was performed with Genomic Mini and Blood Mini isolation kits (A&A Biotechnology) according to the manufacturer's protocol. The microsatellite markers D10S1709 (10q) , D10S1172 (10p) , D22S283 (22q) were selected using the NCBI database , 21. TheRNA isolation and purification was performed using RNA isolation kit (A&A Biotechnology). In the next step samples were transcribed into cDNA with SuperScript II Reverse Transcriptase according to the manufacturer's protocol. Real-Time PCR quantitation was carried out using TaqMan Real-Time PCR Master Mix and TaqMan probes detecting genes which products are involved in DSB repair pathways . 18S rRNA TaqMan probe was included as the reference gene. The parameters for Agilent Technologies Stratagene Mx300SP instrument were 95\u00b0C for 10 minutes, 30 cycles of 95\u00b0C for 15 seconds and 60\u00b0C for 60 seconds.Protein extraction was performed incubating cell pellet with the mixture of RIPA buffer (Sigma) and protease inhibitor cocktail . After concentration measurement, 30 \u03bcg of cell lysates was resolved on 4\u201320% ExpressPluS PAGE Gel . The proteins were then transferred onto PVDF Transfer Membrane using eBlot Protein Transfer device . Membranes were blocked and blotted overnight with primary antibodies recognizing LIG4 and GAPDH . Membranes were then washed and incubated 1 h with secondary anti-mouse antibody conjugated with HRP . The result was visualized using Pierce ECL Western Blotting Substrate and BioRad Universal Hood II with Chemiluminescence System .5 viable cells per well. Cells were cultured with 50 nM BMN673 (Selleckchem), 6.25 \u03bcM TMZ (Sigma Aldrich), BMN673 + temozolomide or vehicle for 48 h followed by the second dose of the compounds and another 72 h of incubation.Normal astrocytes and glioblastoma cells were plated in a 6-well plate at a density of 2 \u00d7 10After the indicated treatments, normal and cancer cells were incubated for 30 min at 37\u00b0C with the mixture of 2 mM Calcein AM and propidium iodide 1 mM diluted in PBS. Fluorescence emitted by stained cells was then observed in an inverted fluorescence microscope .Flow cytometry and staining with propidium iodide and FITC Annexin V (BD Biosciences) was used to assess changes in viability and to track the mechanism of cell death after treatment. Cells were prepared and analyzed according to the FITC Annexin Apoptosis Detection Kit II (BD Biosciences). To analyze the influence of the compounds on glioblastoma and NHA distribution in cell cycle, cells fixed with 70% cold ethanol were stained with propidium iodide with addition of RNase (BD Biosciences) and analyzed. The extent of DNA DSBs measured by phosphorylation of H2A.X histone was obtained using Alexa Fluor 647 Mouse Anti-H2A.X (pS139) antibody after 48 h treatment with the compounds. Fixed cells were washed resuspended in 20 \u03bcl BD Perm/Wash\u2122 buffer and stained for 20 min with H2A.X antibody (5 \u03bcl/test). All the experiments were performed using a FACS Canto II cytometer .Neutral comet assay was performed according to the protocol used in the previous research on cells3 cells were resuspended in 700 \u03bcl of soft agar and plated over 700 \u03bcl of solidified agar underlay on a 12-well plate. After solidifying cell layer was covered with medium (changed weekly). After 2\u20133 weeks colonies were stained with crystal violet (0.5% w/v) and counted under the microscope. Clonogenic efficiency was expressed as percent of untreated control (no. of colonies after treatment vs no. of colonies in control sample \u00d7 100%).To examine clonogenic activity glioblastoma cells were first cultured with drugs or vehicle for 48 h followed by the second dose of the compounds and another 72 h of incubation. After treatment cell viability was determined by staining with trypan blue and 10Glioblastoma H6 cells were transfected with pCMV6-AC-GFP plasmid containing human LIG4 cDNA (OriGene Technologies). The method was performed using Lipofectamine 2000 . GFP-positive cells were sorted 48 h after transfection.Microarray data sets were obtained from NCBI GEO GSE13041). Gene expression profiling was performed as described before \u201330. Micr41. Gene t test. P values lower than 0.05 were considered significant. The synergistic effect of drugs was studied using response additivity approach.Data was accessed in three independent experiments and presented as mean \u00b1 SD. Results were compared using two tailed Student Studies performed on cells derived from surgical specimens were approved by the Ethical Commission of the Medical University of Lodz (no. RNN/194/12/KE) and informed consent was obtained from all patients."} +{"text": "Mobile health applications (app) have shown to be beneficial for chronic disease management. However, few studies assessed older adults\u2019 engagement in tracking self-management activities with app functions and effectiveness on improving their diabetes outcomes. This study investigated tracking patterns of each app function in a graphic-based aging-friendly diabetes self-management app (IMTOP app) and associated the patterns with changes in HbA1c, self-care behavior, diabetes empowerment, and health promotion. The sample included 334 community-dwelling older adults with type 2 diabetes in Taiwan (mean age 64.57 \u00b1 6.64 years) participated in the IMTOP training course that designed to motivate and train older adults with diabetes to use mobile tablets and apps. We performed trajectory analyses using SAS TRAJ procedure to identify distinct classes of individuals following similar longitudinal patterns on absence or presence of weekly app use for each individual app function. The relationships between the app engagement class memberships and 4- and 8-month diabetes health outcomes were assessed using an econometric regression analysis approach. The results showed the degree of app engagement on any single function was significantly and positively correlated with diabetes self-care scale scores . Only the engagement on the blood sugar function had statistically significant association with HbA1c improvements (p < .05). The app use was not associated with diabetes empowerment or health promotion. The study findings suggest any app function engagement significantly improved older adults\u2019 overall self-management but blood sugar tracking is critical to improve HbA1c."} +{"text": "Many adults with IgG subclass deficiency (IgGSD) experience long intervals of frequent/severe respiratory tract infection before IgGSD diagnosis, but reasons for delays in IgGSD diagnoses are incompletely understood. We performed a retrospective study of 300 white adults (ages \u226518 y) with IgGSD including frequency analyses of age at IgGSD diagnosis, duration of frequent/severe respiratory tract infection before IgGSD diagnosis, and age at onset of frequent/severe infection . We performed multivariable regressions on age at diagnosis, infection duration, and age at infection onset using these variables, as appropriate: sex; age at diagnosis; diabetes; autoimmune condition(s); atopy; allergy; corticosteroid use; body mass index; serum immunoglobulin isotype levels; blood lymphocyte subsets; three IgGSD-associated human leukocyte antigen-A and -B haplotypes; and referring physician specialties. Mean age at diagnosis was 50 \u00b1 12 (standard deviation) y (median 50 y (range 19\u201379)). There were 247 women (82.3%). Mean infection duration at IgGSD diagnosis was 12 \u00b1 13 y (median 7 y (range 1\u201366)). Mean age at infection onset was 38 \u00b1 16 y ). Age at infection onset was \u226518 y in 95.7% of subjects. Regressions on age at diagnosis and infection duration revealed no significant associations. Regression on age at infection onset revealed one positive association: age at diagnosis (p <0.0001). We conclude that the median duration of frequent/severe respiratory tract infection in adults before IgGSD diagnosis was 7 y. Older adults may be diagnosed to have IgGSD after longer intervals of infection than younger adults. Duration of frequent/severe respiratory tract infection before IgGSD diagnosis was not significantly associated with routine clinical and laboratory variables, including referring physician specialties. Immunoglobulin (Ig) G subclass deficiency (IgGSD) in adults is characterized by frequent or severe respiratory tract infection, suboptimal IgG response to polyvalent pneumococcal polysaccharide vaccination, female predominance, and increased prevalence of autoimmune conditions \u20133. Many To learn more, we performed a retrospective study of 300 unrelated non-Hispanic whites diagnosed to have IgGSD as adults (ages \u226518 y). We analyzed distributions of ages at IgGSD diagnosis, duration of frequent or severe respiratory tract infection before IgGSD diagnosis, and ages at onset of frequent or severe respiratory tract infection. We also performed multivariable regressions on age at IgGSD diagnosis, duration of frequent or severe respiratory tract infection, and age at onset of frequent or severe respiratory tract infection using these independent routine clinical and laboratory variables, as appropriate: sex; age at diagnosis; diabetes; autoimmune condition(s); atopy; allergy; corticosteroid use; body mass index; serum Ig isotype levels; blood lymphocyte subsets; three human leukocyte antigen (HLA)-A and -B haplotypes associated with IgGSD in adults; and specialties of referring physicians. Herein, we determined that the median duration of frequent/severe respiratory tract in adults before IgGSD diagnosis was 7 y and that the duration of frequent/severe respiratory tract infection before IgGSD diagnosis was not significantly associated with routine clinical and laboratory variables, including referring physician specialties. We discuss our findings in the context of previous reports of IgGSD diagnosis in adults.This work was performed according to the principles of the Declaration of Helsinki . WesternWe studied consecutive unrelated self-identified non-Hispanic white adults ages \u226518 y) referred to a single outpatient referral practice because they had frequent or severe upper or lower respiratory tract infection and were diagnosed to have IgGSD 8 y refer before 2Mycobacterium sp., or human immunodeficiency virus; and incomplete evaluations.We excluded adults with Ig deficiency other than IgGSD , includiWe compiled reports of duration of frequent or severe respiratory tract infection before IgGSD diagnosis. Age at onset of frequent or severe infection was defined as the difference between age at IgGSD diagnosis and reported duration of frequent or severe respiratory tract infection before IgGSD diagnosis.2.We classified diabetes according to the criteria of the American Diabetes Association . AutoimmSerum Ig levels were measured using standard methods before IgG replacement therapy was initiated. We defined mean \u00b1 2 standard deviations (SD) as reference ranges for all Ig measurements . ReferenBlood lymphocyte subsets were measured using flow cytometry . Reference ranges (mean \u00b1 2 SD) are: CD19+ 12\u2013645 cells/\u03bcL; CD3+/CD4+ 359\u20131,519 cells/\u03bcL; CD3+/CD8+ 109\u2013897 cells/\u03bcL; and CD56+/CD16+ 24\u2013406 cells/\u03bcL. Subnormal levels were defined as those below the corresponding lower reference limit.HLA-A and -B types and haplotypes were detected using low-resolution DNA-based typing (polymerase chain reaction/sequence-specific oligonucleotide probe) . ControlThe dataset for analyses consisted of complete observations on 300 adults. All data underlying the findings reported in this work are provided in a Supporting Information file . All ageMean age at diagnosis was 50 \u00b1 12 y (median 50 y (range 19\u201379)). There were 247 women (82.3%). Otolaryngologists, primary care physicians, rheumatologists, and pulmonologists referred 29.3%, 28.7%, 25.0%, and 9.3% of adults, respectively (92.3% in aggregate). Other referring physicians were endocrinologists, gastroenterologists, neurologists, infectious disease specialists, gynecologists, and cardiologists (in decreasing order).2 (range 16.3\u201368.6).Thirty-one adults (10.3%) had type 2 diabetes. One hundred and fifteen adults (38.3%) were diagnosed to have an autoimmune condition(s). Atopy, other allergy, and corticosteroid therapy occurred in 24.0%, 27.3%, and 18.7% of adults, respectively. Median body mass index was 27.9 kg/mSubnormal blood levels of CD19+, CD3+/CD4+, CD3+/CD8+, and CD56+/CD16+ cells were observed in 1.0%, 5.0%, 1.7%, and 3.3% of adults, respectively. Elevated blood levels of CD19+, CD3+/CD4+, CD3+/CD8+, and CD56+/CD16+ cells were observed in 2.3%, 1.2%, 3.7%, and 1.0% of adults, respectively.IgG immunophenotypes are displayed in These data were normally distributed . The ageThese data were not normally distributed . Mean duAge at onset of frequent or severe respiratory tract infection data were not normally distributed . Mean agWe performed a backward stepwise variable regression on age at IgGSD diagnosis using these independent variables: sex; diabetes; autoimmune condition(s); atopy; allergy; corticosteroid use; body mass index; serum Ig isotype levels; blood lymphocyte subsets; three HLA-A and -B haplotypes; and referring physician specialties. This regression revealed no significant associations.We performed a backward stepwise variable regression on duration of infection before IgGSD diagnosis using these independent variables: sex; age at diagnosis; diabetes; autoimmune condition(s); atopy; allergy; corticosteroid use; body mass index; serum Ig isotype levels; blood lymphocyte subsets; three HLA-A and -B haplotypes; and referring physician specialties. This regression revealed no significant associations.We performed a backward stepwise variable regression on age at onset of frequent or severe respiratory tract infection using these independent variables: sex; age at diagnosis; diabetes; autoimmune condition(s); atopy; allergy; corticosteroid use; body mass index; serum Ig isotype levels; blood lymphocyte subsets; three HLA-A and -B haplotypes; and referring physician specialties. This regression revealed a single positive association: age at diagnosis (p <0.0001). This regression accounted for 42.2% of the deviance of age at onset of infection (ANOVA p of regression <0.0001).The mean age (range) at IgGSD diagnosis in the present adults (50 y (19\u201379)) is consistent with those of three other adult IgGSD cohorts from The Netherlands and US: 49 y (33\u201370) ; 51 y . In a cohort of English adults, the median duration of frequent or severe respiratory tract infection before diagnosis of IgGSD or specific antibody deficiency was 11 y . Most KoFrequent or severe respiratory tract infection is the most common manifestation of undiagnosed IgGSD ,18\u201320. BDiabetes was diagnosed in more than 10% of the present adults with IgGSD, a proportion similar to that of adults ages \u226518 y with diabetes in the general Alabama population (12%) . DiabetePhysicians of different specialties treat patients with frequent or severe respiratory tract infection. Otolaryngologists, primary care physicians, rheumatologists, and pulmonologists referred 92.3% of the present adults, consistent with results of a previous study of adults with IgGSD or common variable immunodeficiency . Pre-refNinety-six percent of the present adults reported that they experienced frequent or severe respiratory tract infection at ages \u226518 y, suggesting that IgGSD in most adults becomes clinically manifest in adulthood. Almost half of the present adults reported that they first experienced frequent or severe respiratory tract infection at ages \u226435 y. Our multivariable regression on age at onset of infection before IgGSD diagnosis revealed a significant positive association with age at IgGSD diagnosis only. This suggests that older adults are diagnosed to have IgGSD after longer intervals of frequent or severe respiratory tract infection than younger adults, after adjustment for other variables.The principal significance of this study is that many of the present adults with frequent or severe respiratory tract infection experienced a delay of many months or years before the underlying cause of their infections was diagnosed, yet most of them had repeated healthcare encounters for treatment of respiratory tract infection before they were evaluated with Ig measures . ConsequA strength of this study is that our sample size provides statistical power to detect significant independent associations with age at IgGSD diagnosis, duration of frequent or severe respiratory tract infection before IgGSD diagnosis, and age at onset of frequent or severe respiratory tract infection in multivariable models. Limitations of this study include lack of serum Ig levels measured in the present adults long before IgGSD diagnosis and possible inaccuracy of reports of duration of frequent or severe respiratory tract infection before IgGSD diagnosis. Another limitation is the lack of observations on a) ostensibly healthy, age- and sex-matched control subjects from the general non-Hispanic white population and b) other control subjects who had frequent or severe respiratory tract infection without evidence of IgGSD or other antibody deficiency. Evaluation of other heritable or acquired factors or health care delivery features that may influence duration of frequent or severe respiratory tract infection in adults before IgGSD diagnosis and morbidity associated with delayed diagnosis of IgGSD was beyond the scope of this study.Median duration of frequent or severe respiratory tract infection in adults before IgGSD diagnosis was 7 y. Older adults may be diagnosed to have IgGSD after longer intervals of infection than younger adults. Duration of frequent or severe respiratory tract infection before IgGSD diagnosis was not significantly associated with routine clinical and laboratory variables, including referring physician specialties.S1 File(XLSX)Click here for additional data file."} +{"text": "Many NCS proteins ) form functional dimers under physiological conditions. The dimeric NCS proteins have similar amino acid sequences (50% homology) but each bind to and regulate very different physiological targets. Retinal recoverin binds to rhodopsin kinase and promotes Ca2+-dependent desensitization of light-excited rhodopsin during visual phototransduction. The guanylyl cyclase activating proteins (GCAP1\u20135) each bind and activate retinal guanylyl cyclases (RetGCs) in light-adapted photoreceptors. VILIP1 binds to membrane targets that modulate neuronal secretion. Here, I review atomic-level structures of dimeric forms of recoverin, GCAPs and VILIP1. The distinct dimeric structures in each case suggest that NCS dimerization may play a role in modulating specific target recognition. The dimerization of recoverin and VILIP1 is Ca2+-dependent and enhances their membrane-targeting Ca2+-myristoyl switch function. The dimerization of GCAP1 and GCAP2 facilitate their binding to dimeric RetGCs and may allosterically control the Ca2+-dependent activation of RetGCs.Neuronal calcium sensor (NCS) proteins are EF-hand containing Ca The Ca2+-induced dimerization of recoverin enhances membrane binding by creating a dual pronged myristoyl anchor compared to the affinity of a monomeric Ca2+-myristoyl switch ) is equal to the square of the dissociation constant of the monomer: Kd(dimer) = Kd(monomer)2 = (10\u20134 M)2 = 10\u20138 M. Thus, the membrane binding affinity of Ca2+-myristoyl switch proteins is predicted here to be dramatically enhanced by the combined effect of both Ca2+-binding and protein dimerization. Dimerization of Ca2+-myristoyl switch proteins may also entropically enhance its binding to dimeric protein targets, as was suggested for the binding of dimeric Ca2+-bound recoverin to dimeric rhodopsin that each weaken dimerization also abolish activation of the cyclase , which may help explain the steep CaJA wrote and conceived the entire manuscript.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Decades of work have shown that diabetes increases the risk of heart disease and worsens clinical outcomes after myocardial infarction. Because diabetes is an absolute contraindication to heart transplant, cell therapy is increasingly being explored as a means of improving heart function for these patients with very few other options. Given that hyperglycemia promotes the generation of toxic metabolites, the influence of the key detoxification enzyme glyoxalase 1 (Glo1) on chronic hyperglycemia induced heart explant-derived cell (EDC) dysfunction was investigated.Methods: EDCs were cultured from wild type C57Bl/6 or Glo1 over-expressing transgenic mice 2 months after treatment with the pancreatic beta cell toxin streptozotocin or vehicle. The effects of Glo1 overexpression was evaluated using in vitro and in vivo models of myocardial ischemia.Results: Chronic hyperglycemia reduced overall culture yields and increased the reactive dicarbonyl cell burden within EDCs. These intrinsic cell changes reduced the angiogenic potential and production of pro-healing exosomes while promoting senescence and slowing proliferation. Compared to intra-myocardial injection of normoglycemic cells, chronic hyperglycemia attenuated cell-mediated improvements in myocardial function and reduced the ability of transplanted cells to promote new blood vessel and cardiomyocyte growth. In contrast, Glo1 overexpression decreased oxidative damage while restoring both cell culture yields and EDC-mediated repair of ischemic myocardium. The latter was associated with enhanced production of pro-healing extracellular vesicles by Glo1 cells without altering the pro-healing microRNA cargo within.Conclusions: Chronic hyperglycemia decreases the regenerative performance of EDCs. Overexpression of Glo1 reduces dicarbonyl stress and prevents chronic hyperglycemia-induced dysfunction by rejuvenating the production of pro-healing extracellular vesicles. Over the past decade, heart-derived cell therapy has emerged as a promising means to promote therapeutic regeneration In this study, we use a transgenic murine model of Glo1 overexpression to critically evaluate the effect of chronic hyperglycemia on EDC-derived EVs. EVs are cell-derived vesicles enclosing proteins and nucleic acids that are naturally secreted as a means of cell-to-cell communication All animal protocols were reviewed and approved by the University of Ottawa Animal Care Committee. Male and female murine cardiac tissue was obtained from 3.4\u00b11.3 month old wild-type C57Bl/6 (WT) or C57Bl/6-PEP8-Glo1 (Glo1) transgenic mice Hyperglycemia was induced by intraperitoneal injection of streptozotocin in 0.05 M sodium citrate (Fisher Scientific). Non-hyperglycemic control mice received equal volumes of 0.05 M sodium citrate. Fasting blood glucose and glycated hemoglobin (HbA1c) measurements were measured prior to sacrifice for enzyme- linked immunosorbent assay .Bioenergetic determinations were performed using Agilent-Seahorse XF24 analyzer Cell growth within 1% oxygen 1% serum 5% glucose conditions was quantified using a colorimetric assay (Dojindo) with confirmatory manual haemocytometer cell counts. Hydrogen peroxide is a major form of reactive oxygen species (ROS) and its level was assayed in cell at 5% oxygen with 20% serum physiological glucose conditions using 2',7-dichlorofluorescein diacetate fluorescent emission . Cell senescence within 5% oxygen 1% serum physiological glucose conditions was quantified using senescence-associated \u03b2-galactosidase activity . The percentage of \u03b2-galactosidase cells was quantified in five random fields per cell line assayed. The number of apoptotic cells within 1% oxygen 1% serum physiological glucose conditions cultured with 0.1\u00b5M staurosporine (Millipore Sigma) was quantified using flow cytometry for phycoerythrin-Annexin V (PE-Annexin V) and 7-Aminoactinomycin D .5 EDCs or vehicle injected into the infarct border zone using echocardiographic guidance The ability of transplanted EDCs to promote cardiac function after permanent left coronary artery (LCA) ligation was evaluated using male murine EDCs injected into female wild-type C57 mice Conditioned medium was prepared from confluent cultures after 48 hours within 1% oxygen, and 1% serum under physiological glucose conditions in the presence and absence of 20 \u03bcM GW4869 (Millipore Sigma). The angiogenic potential of EDC conditioned media was evaluated using a cytokine depleted matrigel assay Conditioned medium was collected after 48 hours of culture within 5% oxygen and 1% exosome- free serum under physiological glucose conditions. EVs were isolated using ExoQuick-TC exosome precipitation solution (System Biosciences) for EV tracking analysis . Micro and total RNA were extracted (Qiagen) and a small RNA library was prepared (Lexogen) for RNASeq evaluation of expression profiles (Illumina NextSeq500). Quality assessment was performed using FastQC (Babraham Institute) and reads were trimmed with a minimum sequence length of 18bp for mapping using bowtie (v1.1.2) aligned to the mus musculus mature microRNA (miRNA) sequences. Differential expression analysis was performed using the negative binomial model of read counts implemented in the DESeq2 R library. Principal component analysis was applied to the matrix of gene expression values (read counts) to identify the major components of gene expression variation. Hierarchical clustering was calculated using Euclidian distance between rlog-transformed normalized count values for all transcripts.All procedures and analyses were performed blinded to animal or cell identity. All data are presented as mean \u00b1 SEM. To determine if differences existed within groups, data was analyzed by a one-way or two-way ANOVA, as appropriate. If such differences existed, Sidak's multiple comparisons test was used to determine the group(s) with the difference(s) . A final value of P\u22640.05 was considered significant for all analyses.EDCs are the early cell product collected from plated cardiac biopsies prior to antigenic selection Unexpectedly, cell culture yields from control normoglycemic tissue were decreased therapy and uniformly showed these products to be safe with promising hints of efficacy We found that chronic hyperglycemia decreases both EDC culture yields and the ability of these cells to withstand oxidative stress or senescence. Intramyocardial injection after cardiac damage also demonstrated that hyperglycemia reduces the angiogenic, mitotic and anti-scarring properties of EDCs. Unlike other comorbidities (such as advanced age or hypertension) Several years ago, we demonstrated that \u201chigh glucose\u201d culture conditions 25 mM) significantly impacted the pro- angiogenic capacity of non-diabetic EDCs mM signiFrom previous work, we suspected that methylglyoxal detoxification might attenuate the adverse effects of chronic hyperglycemia In the course of the study, we unexpectedly found that our inbred Glo1 mice displayed impaired cellular metabolic characteristics which may be related to secondary effects of high levels of Glo1 overexpression such as altered glutathione and/or NADH redox potentials, or the shunting of glycolytic 3-carbon metabolites into the methylglyoxal pathway, away from oxidative pathways. Despite this tendency, Glo1 overexpression markedly attenuated the impact of hyperglycemia on post infarct repair by increasing EV production and endogenous repair which suggests these adverse metabolic derangements were more than compensated by the cytoprotective effects conferred by prevention of methylglyoxal overload. As such, we provide the first proof that reducing methylglyoxal content during chronic hyperglycemia prevents the adverse effects of hyperglycemia.We also noted a marked disparity in the EV nanoparticle content within media conditioned by normoglycemic Glo1 or WT mice . Despite this, equivalent degrees of post infarct function and scar burden were seen. This observation supports our hypothesis that cardiac cell transplant outcomes are dependent on exposure to a potent paracrine cell product and, once a stimulation threshold is achieved, simply increasing the number of cells retained or EVs provided has no further benefits in vivo study design employed a non-diabetic recipient to avoid confounding effects from recipient hyperglycemia but the regenerative performance of cells within the hyperglycemic host deserves further study as cell treatment outcomes are clearly leveraged on endogenous repair mechanisms. Finally, panning through ventricular lysate failed to demonstrate meaningful persistence of transplanted cells. Previous work has shown that long-term persistence of transplanted cells is very modest In terms of study limitations, the STZ model chosen reflects insulin deficient hyperglycemia while the majority of patients potentially in need of cell therapy are type 2 diabetics with both hyperglycemia and insulin resistance. Although the STZ model provides an unbiased platform to observe the effects of chronic hyperglycemia, further work to extend this data to patients with type 2 diabetes and ischemic cardiomyopathy are needed. Our"} +{"text": "Hexanchus griseus) in response to environmental changes experienced during diel vertical migrations. In the subtropical waters off Hawai\u2018i, sixgill sharks undertook pronounced diel vertical migrations and spent considerable amounts of time in cold (5\u20137\u00b0C), low oxygen conditions (10\u201325% saturation) during their deeper daytime distribution. Further, sixgill sharks spent the majority of their deeper daytime distribution with intramuscular temperatures warmer than ambient water temperatures, thereby providing them with a significant thermal advantage over non-vertically migrating and smaller-sized prey. Sixgill sharks exhibited relatively high rates of activity during both shallow (night) and deep (day) phases and contrary to our predictions, did not reduce activity levels during their deeper daytime distribution while experiencing low temperature and dissolved oxygen levels. This demonstrates an ability to tolerate the low oxygen conditions occurring within the local oxygen minimum zone. The novel combination of biologging technologies used here enabled innovative in situ deep-sea natural experiments and provided significant insight into the behavioral and physiological ecology of an ecologically important deepwater species.Diel vertical migration is a widespread behavioral phenomenon where organisms migrate through the water column and may modify behavior relative to changing environmental conditions based on physiological tolerances. Here, we combined a novel suite of biologging technologies to examine the thermal physiology (intramuscular temperature), fine-scale swimming behavior and activity of bluntnose sixgill sharks ( Hexanchus griseus; hereafter referred to as sixgill sharks) undergo a distinct diel vertical migration descending to depths below 500 m during the day and ascending to 200\u2013350 m during the night and encounter a wide range of temperatures and and Hexa . Sixgil ) compared to smaller species [crit) [2) at which blood is 50% saturated (P50) [Given the greater energy yield of aerobic metabolism, we expect anaerobic metabolism to merely support activity levels above routine metabolic rates for short periods of time, as opposed to completely supporting metabolic expenditure throughout the daytime distribution under hypoxic conditions . However species \u201378. In as [crit) ,18. For s [crit) ,18. In as [crit) ,18,72,79) and thePrionace glauca) and ocean sunfish (Mola mola) make frequent ascents to surface waters well before intramuscular temperatures reach equilibrium with ambient water temperatures experienced at depth and employ variable heat-transfer coefficients where intramuscular temperatures warm more quickly than it cools [Thunnus obesus) [The enhanced thermal inertia of sixgill sharks may confer ecological and physiological advantages at cold temperatures as suggested for fishes with endothermic capacities. Endothermic fishes have evolved specialized anatomical and physiological mechanisms for retaining metabolic heat enabling higher levels of activity for longer durations when diving below the mixed layer to exploit deep forage resources more effectively than ectothermic species \u201383. Howeit cools ,84, possit cools and bige obesus) ,32. In c obesus) ,89. We s obesus) , low actODBA had many extreme values that needed to be accommodated in both modeling approaches. This issue could be resolved in HMM analyses by estimating the state-dependent densities nonparametrically, particularly for the high-activity state ,50,90. Fin situ deep-sea natural experiments, thus improving our understanding of the biological response of deepwater species to contemporary and future ocean conditions.The novel combination of biologging technologies used in this study provides significant insight into the behavioral and physiological ecology of a deepwater species. The different modeling approaches provide a robust framework for empirically testing the influence of environmental drivers on the activity of sixgill sharks across diel vertical migrations and empirically demonstrated their tolerance of cold, low oxygen conditions in deepwater habitats. Obtaining similar data in regions with more variable and/or lower dissolved oxygen content may help define the oxygen tolerance for this species and resolve the relative impacts of cold temperatures and low oxygen on behavior ,72. SignS1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file.S3 Fig(PDF)Click here for additional data file.S4 Fig(PDF)Click here for additional data file.S5 Fig(PDF)Click here for additional data file.S6 Fig(PDF)Click here for additional data file.S7 Fig(PDF)Click here for additional data file.S8 Fig(PDF)Click here for additional data file.S9 Fig(PDF)Click here for additional data file.S1 Table(PDF)Click here for additional data file.S2 Table(PDF)Click here for additional data file.S3 Table(PDF)Click here for additional data file.S4 Table(PDF)Click here for additional data file.S5 Table(PDF)Click here for additional data file.S6 Table(PDF)Click here for additional data file."} +{"text": "Air pollution (AirPoll) is among the leading human mortality risk factors and yet little is known about the molecular mechanisms of this global environmental toxin. Our recent studies using mouse models even showed genetic variation and sex can alter biological responses to air pollution. To expand genetic studies of AirPoll toxicity throughout the lifespan, we introduced Caenorhabditis elegans as a new AirPoll exposure model which has a short lifespan, high throughput capabilities and shared longevity pathways with mammals. Acute exposure of C. elegans to airborne nanosized AirPoll matter (nPM) caused similar gene expression changes to our prior findings in cell culture and mouse models. Initial C. elegans responses to nPM included antioxidant, inflammatory and Alzheimer homolog genes. The magnitude of changes was dependent on the developmental stage of the worms. Even short term exposure of C. elegans to nPM altered developmental and lifespan hormetic effects, with pathways that included skn-1/Nrf family antioxidant responses. We propose C. elegans as a new and complementary model for mouse and cultured cells to study AirPoll across the lifespan. Future chronic nPM exposure and high throughput genetic screening of C. elegans can identify other major regulators of the developmental and lifespan effects of air pollution. This work was supported by grants R01AG051521 (CEF); R21AG05020 (CEF); Cure Alzheimer\u2019s Fund (CEF); R01GM109028 (SPC), F31AG051382 (HMD) and T32AG000037 (HMD), T32AG052374 (AH)."} +{"text": "Mycobacterium tuberculosis isolates from extrapulmonary sites\u201d [1].This article describes the whole genome sequencing data from 5 extrapulmonary tuberculosis clinical isolates. The whole genome sequencing was carried out on Illumina MiSeq platform to identify single nucleotide variations (SNVs) associated with drug resistance. A total of 214 SNVs in the coding and promoter regions were identified in the whole genome sequencing analysis. Among the identified SNVs, 18 SNVs were identified in genes known to be associated with first and second line drug resistance. The data is related to the research article \u201cWhole genome sequencing of Specifications TableValue of the data\u2022M. tuberculosis clinical isolates from extra pulmonary sitesThis data provides insight into the genomic profiles of \u2022Lineage-specific SNVs identified in whole genome sequencing allows accurate strain typing and provided the information of lineage distribution of EPTB isolates\u2022The data also provided information on SNVs associated with conferring resistance to anti-tubercular drugs\u2022M. tuberculosis isolated from different infection sitesSince genomic profiles of EPTB isolates remains largely unexplored, this data would add value to our current knowledge on genomes of 1) . Three of these 157 genes are associated with drug resistance show promoter region SNVs in all of the 5 isolates A. A scieSupplementary Table 1B) . Three oisolates B.Fig. 1 using Burrows-Wheeler Alignment Tool (BWA version-0.7.15)"} +{"text": "In 2003, the Assisted Living Workgroup (ALW) published quality improvement recommendations for states\u2019 regulations, including 26 regarding staffing/workforce. We reviewed states\u2019 2003 and current regulations to identify the presence of ALW standards. Over half of states\u2019 regulations reflect 7 of the 26 staffing/workforce recommendations. Those most often added after 2003 concern criminal background checks, with a 58.8 percent increase in states that added federal background checks and use of criminal background checks to inform hiring. At least 40 states\u2019 regulations reflect the ALW recommendations for administrator and direct care staff training. Very few states require staff performance evaluations (n=13), human resource policies to improve retention (n=1), or management practices to improve retention (0). The 10 ALW recommendations concerning staff who administer medications have been adopted by fewer than 23 states. These findings can inform future policy analysis and research on staffing/workforce in assisted living communities."} +{"text": "Lung cancer is the leading cause of cancer-related deaths in Asian Americans. Low-dose computed tomography lung cancer (LDCT) screening is an effective way to decrease lung cancer mortality. This study aimed to examine the difference in LDCT screening eligibility among Asian American subgroups. The National Health Interview Survey data (2006-2016) was analyzed. The U.S. Preventive Services Task Force guideline was used to determine the LDCT eligibility. A higher and statistically significant proportion of current Filipino smokers (35.4%) met LDCT screening eligibility criteria compared to Chinese (26.5%) and other Asian smokers (22.7%) (p=0.02). Hierarchical logistic regression results further showed that Filipino were more likely to meet LDCT screening criteria than other Asian while adjusting demographics . The differences in LDCT screening eligibility no longer existed after additionally adjusting socioeconomic factors as well as perceived health status. Future targeted outreach and intervention research is needed for Filipinos with lower socioeconomic status."} +{"text": "We identified a Eurasian-origin influenza A(H8N4) virus in North America by sampling wild birds in western Alaska, USA. Evidence for repeated introductions of influenza A viruses into North America by migratory birds suggests that intercontinental dispersal might not be exceedingly rare and that our understanding of viral establishment is incomplete. Research of and surveillance for influenza A viruses in wild birds inhabiting western Alaska have consistently provided support for the exchange of viruses between East Asia and North America via Beringia (Anseriformes spp.) and 401 environmental fecal samples from monospecific flocks of either emperor geese (Chen canagica) or glaucous-winged gulls (Larus glaucescens) within and around Izembek NWR. Samples were deposited into viral transport media, placed in dry shippers charged with liquid nitrogen within 24 h, shipped, and stored frozen at \u221280\u00b0C before laboratory analysis. We screened samples for the influenza A virus matrix gene and subjected them to virus isolation; resultant isolates were genomically sequenced in accordance with previously reported methods by incorporating sequence information for representative reference sequences from avian-origin influenza A virus isolates from Eurasia and North America using the general time-reversible plus invariant sites (G+I) model with 1,000 bootstrap replications.We queried sequence information for the complete coding region of each gene segment of A/northern pintail/Alaska/UGAI16-3997/2016(H8N4) against the GenBank database to identify strains sharing >99% nt identity to those of >1 isolates recovered from wild and domestic birds sampled in East Asia during 2006\u20132016 , isolated from a sample collected from a hunter-harvested duck on September 6, 2016, shared 006\u20132016 Table. Tsegments Table. Aellation Table.>99; Phylogenetic analyses strongly supported structuring of tree topologies into major clades by continental affiliation of reference sequences suggest that viral dispersal between East Asia and North America might not be exceedingly rare. Thus, a lack of selective advantage for comparatively rare foreign-origin influenza A viruses, purifying selection for endemic viruses, or both might be important mechanisms regulating the establishment of these viruses within the wild bird reservoir. Therefore, additional research directed toward understanding selection pressures regulating the establishment of these viruses might provide useful inference for informing surveillance and response activities for economically costly or potentially pandemic foreign-origin viruses in wild birds inhabiting North America.During 2010\u20132016, research and surveillance for influenza A viruses in wild birds inhabiting North America have provided evidence for the intercontinental dispersal of the following 4 viral genome constellations between Eurasia and North America: H16N3 (>1 gene segments with A/northern pintail/Alaska/UGAI16\u20133997/2016(H8N4), March 12, 2018; unrooted maximum-likelihood phylogenetic trees for the complete coding regions of the gene segments for influenza A virus strain A/northern pintail/Alaska/UGAI16\u20133997/2016(H8N4).Viral isolates sharing \u226599% nt identity at"} +{"text": "This study shows that mRNA or nuclear protein levels of the ZNF217 significantly correlate with Oncotype DX\u00ae Recurrence Score.We assessed mRNA and protein expression levels of the ZN217 oncogene in 17 clinical FFPE ER-positive invasive breast cancer specimens with known (low or high) Oncotype DX Thus, a more precise method for stratifying patients based on their prognosis and for predicting their response to therapy remains needed.Breast cancer (BC) is the most frequent cancer among women. Expression of Estrogen Receptor \u03b1 (ER\u03b1) is found in 60\u201380% of BC patients, and allows an accurate prediction of response to endocrine therapy (ET). However, between 10 and 50% of ER\u00ae (ODX) genomic assay tests for the expression of 21 genes and calculates a Recurrence Score (RS), which predicts the risk of distant disease recurrence in ER+ BC. A high RS value indicates a poor prognosis and a higher probability of distant recurrence at 10 years in patients treated with adjuvant ET low-risk (<18) or high-risk (>31) ODX RS; (ii) sufficient tissue for both ZNF217 immunohistochemistry (IHC) and the mean value was used as a cutoff. ZNF217 mRNA levels (<4) or low percentage of IHC stained nuclei (<5%); (ii) ZNF217 nuclear staining and ZNF217 mRNA levels were significantly associated with ODX RS; (iii) combining both IHC analysis and ZNF217 mRNA levels allowed the stratification of the samples with a better accuracy, with 100 and 80%, respectively, of low-risk ODX RS and high-risk ODX RS correctly classified and significantly association with ZNF217 expression levels (P = 0.002). Strikingly, two high ODX RS specimens displaying the highest ZNF217 mRNA levels (9.2 and 22.5) also displayed the highest ZNF217 IHC staining (70\u201380%) and pejorative clinical record .After approval by the Institutional Review Board, the pathology database of the Montefiore Medical Center was searched to identify ER+ BC. Supporting recent observations indicate that ZNF217 expression levels also predict neoadjuvant ET response in these patients .PC and SF conceived the study. SF supervised and analyzed the IHC experiments performed by JA. OL supervised and analyzed the RTQ-PCR experiments performed by CL. PC and SF co-analyzed the data. PC wrote the manuscript.SF served in an expert advisory panel for Genomic Health. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The human intestinal microbiota contains a vast community of microorganisms increasingly studied using high-throughput DNA sequencing. Standardized protocols for storage and DNA extraction from fecal samples have been established mostly for bacterial microbiota analysis. Here, we investigated the impact of storage and DNA extraction on bacterial and fungal community structures detected concomitantly.later\u00ae and subjected to 2 extraction protocols with mechanical lysis: the Powersoil\u00ae MoBio kit or the International Human Microbiota Standard (IHMS) Protocol Q. Libraries of the 12 samples targeting the V3-V4 16S and the ITS1 regions were prepared using Metabiote\u00ae (Genoscreen) and sequenced on GS-FLX-454. Sequencing data were analysed using SHAMAN (http://shaman.pasteur.fr/). The bacterial and fungal microbiota were compared in terms of diversity and relative abundance.Fecal samples from healthy adults were stored at -80\u00b0C as such or diluted in RNAlater\u00ae impacted (downward trend) the relative abundances of 7/41 bacterial and 6/40 fungal taxa, while extraction impacted randomly 18/41 bacterial taxa and 1/40 fungal taxon.We obtained 171869 and 199089 quality-controlled reads for 16S and ITS, respectively. All 16S reads were assigned to 41 bacterial genera; only 52% of ITS reads were assigned to 40 fungal genera/section. Rarefaction curves were satisfactory in 3/3 and 2/3 subjects for 16S and ITS, respectively. PCoA showed important inter-individual variability of intestinal microbiota largely overweighing the effect of storage or extraction. Storage in RNAlater\u00ae moderately impacts bacterial or fungal community structures, while extraction significantly influences the bacterial composition. For combined bacterial and fungal intestinal microbiota analysis, immediate sample freezing should be preferred when feasible, but storage in RNAlater\u00ae remains an option under unfavourable conditions or for concomitant metatranscriptomic analysis; and extraction should rely on protocols validated for bacterial analysis, such as IHMS Protocol Q, and including a powerful mechanical lysis, essential for fungal extraction.Our results showed that RNA Using Ps, 8.1%) .later\u00ae; PowerSoil\u00ae MoBio kit), an increased diversity was found with all 40 fungal genera/section detected and 6/40 fungal taxa and 1 more fungal taxa significantly impacted by storage condition in terms of relative abundance (later\u00ae prior freezing (Cryptococcus), which was detected only from samples stored with RNAlater\u00ae for both bacterial and fungal analysis but only 1/40 fungal (Debaryomyces) genera (Roseburia and Sutterella) and 3 fungal additional taxa statistically different according to the extraction protocol genera . Of noteprotocol . Among aprotocol , S5 Fig.later\u00ae and of two well-known extraction protocols on the i.e., all taxa were detected by both methods). Although the number of subjects in our study was low, our results were in agreement with that of other authors, who highlighted the impact of extraction protocols on the assessment of bacterial microbiota or RNAlater\u00ae dilution before freezing [square]) and two extraction protocols (IHMS Protocol Q [dark color] and PowerSoil\u00ae MoBio kit [light color]) are presented. Sixty-five percent and 82% of between-sample variations were explained by the first two PC1 and PC2 axis for bacterial and fungal PCoA analyses, respectively.(TIF)Click here for additional data file.S4 FigBoxplot comparison, at individual level, of log2-abundance of bacterial (A) and fungal (B) taxa according to storage condition. Bacterial diversity was assessed at genus level using 16S rRNA gene ultra-deep sequencing (454 technology) and fungal diversity at genus or section level using ITS1 ultra-deep sequencing. Boxplot of log2-abundance of taxa significantly different according to storage condition (RNAlater\u00ae dilution before freezing [dark colors] vs. within two-hours freezing without additive [light colors]) are presented separately for each individual . Significant differences observed at individual level are indicated using colored asterisks placed above/below boxplots. Significant differences observed at general level are indicated using black asterisks attached to the genera/sections names.(TIF)Click here for additional data file.S5 FigBoxplot comparison, at individual level, of log2-abundance of bacterial (A) and fungal (B) taxa according to extraction protocol. Bacterial diversity was assessed at genus level using 16S rRNA gene ultra-deep sequencing (454 technology) and fungal diversity at genus or section level using ITS1 ultra-deep sequencing. Boxplot of log2-abundance of taxa significantly different according to extraction protocol (PowerSoil\u00ae MoBio kit [dark colors] vs. IHMS Protocol Q [light colors]) are presented separately for each individual . Significant differences observed at individual level are indicated using colored asterisks placed above/below boxplots. Significant differences observed at general level are indicated using black asterisks attached to the genera/sections names.(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 TableAlpha diversity measurements for bacterial (A) and fungal (B) metagenomic analyzes of 3 fecal samples using 2 storage and 2 extraction conditions.(DOCX)Click here for additional data file.S3 TableAbundance Fold Change of bacterial (A) and fungal (B) taxa significantly different according to storage condition at general level.(DOCX)Click here for additional data file.S4 TableAbundance Fold Change of bacterial (A) and fungal (B) taxa significantly different according to storage condition at individual level.(DOCX)Click here for additional data file.S5 TableAbundance Fold Change of bacterial (A) and fungal (B) taxa significantly different according to extraction protocol at general level.(DOCX)Click here for additional data file.S6 TableAbundance Fold Change of bacterial (A) and fungal (B) taxa significantly different according to extraction protocol at individual level.(DOCX)Click here for additional data file."} +{"text": "Recommendations to conceptualize sexual orientation as a continuum and as multidimensional rather than one dichotomous variable have been largely unexplored in sexual minority older adults, including how these dimensions might differ by age and gender. In this study, participants indicated their sexuality using three continua representing (1) attraction in general, (2) emotional attraction, and (3) physical attraction. Possible responses ranged from 0=exclusively opposite sex to 7=exclusively same sex. The current sample included 187 participants self-identifying their sexual attraction in general as not exclusively to the opposite sex. Age groups were 50-55 (n=56), 56-64 (n=84), and 65-86 (n=47) years. MANOVA results indicated a significant multivariate age group by gender interaction (p=.040) that was significant for all three attraction variables---attraction in general (p=.035), emotional attraction (p=.010), and physical attraction (p=.029). In the 50-55 age group, the average response for physical attraction was closer to exclusively same sex for men than for women. For the 56-64 age group, the average response for attraction in general and emotional attraction was closer to exclusively same sex for women than men. Among those 65+, women responded closer to exclusively same sex than men only for emotional attraction. Gender differences on all three sexual attraction continua were not consistent across age groups, which may reflect a more fluid and complex understanding of sexuality in older LGB adults. Future studies should consider using multidimensional and continuous variables when measuring sexual orientation."} +{"text": "Approximately 85% of NH3 emissions are estimated to come from agricultural nonpoint sources. We suspect a strong seasonal pattern in NH3 emissions; however, current NH3 emission inventories lack intra-annual variability. Annually averaged NH3 emissions could significantly affect model-predicted concentrations and wet and dry deposition of nitrogen-containing compounds. We apply a Kalman filter inverse modeling technique to deduce monthly NH3 emissions for the eastern U.S. Final products of this research will include monthly emissions estimates from each season. Results for January and June 1990 are currently available and are presented here. The U.S. Environmental Protection Agency (USEPA) Community Multiscale Air Quality (CMAQ) model and ammonium (NH4) wet concentration data from the National Atmospheric Deposition Program (NADP) network are used. The inverse modeling technique estimates the emission adjustments that provide optimal modeled results with respect to wet NH4 concentrations, observational data error, and emission uncertainty. Our results suggest that annual average NH3 emissions estimates should be decreased by 64% for January 1990 and increased by 25% for June 1990. These results illustrate the strong differences that are anticipated for NH3 emissions.Significant uncertainty exists in magnitude and variability of ammonia (NH"} +{"text": "Dear Editor,n\u2009=\u200913/87) of included patients eventually had a microbiologically proven GAS NSTI. This was a major limitation and early identification of patients with a high probability of GAS-associated NSTIs would thus be crucial for further studies evaluating similar interventions.Necrotizing soft tissue infections (NSTIs) are a heterogeneous group of devastating diseases involving a wide variety of microorganisms and affecting different body areas. The need for individualized treatment strategies has been recently put forward in a prospective cohort study of 402 patients in which group A streptococcus (GAS) infections were associated with more frequent septic shock . Early iA secondary analysis of a retrospective cohort including 224 patients admitted to our center for NSTI between 2006 and 2017 was conducted and 97% [86\u2013100] had a GAS infection, which was monomicrobial in 39 17%) cases. Overall, 134 (59.8%) patients were admitted to the intensive care unit during their stay, of whom 113 during the first 24\u2009h. Ninety-one (41%) patients presented with shock , and 89 (40%) required mechanical ventilation. Sixty days after admission, 51 (23%) patients had died, including 10 (17%) with GAS, and 41 (25%) with non-GAS infections (7% cases.In conclusion, we retrospectively identified two simple and available upon admission clinical predictors of GAS documentation among a large cohort of surgically proven NSTIs. Our results show that NSTI patients with pre-existing immunodeficiency or an abdominal infection have a low probability of GAS infection and might thus not be suitable for inclusion in a trial assessing the effect of GAS-specific interventions. Such findings need to be assessed in a validation cohort in order to reinforce their generalizability. Improving identification upon admission of a subgroup of patients with a higher prevalence of GAS infection might help design future prospective trials aimed at assessing personalized treatment strategies ."} +{"text": "The Baltimore Longitudinal Study of Aging (BLSA), an ongoing continuous enrollment cohort study of normative aging established in 1958 currently conducted by the NIA Intramural Research Program, began including women in 1978. To date, nearly 1200 women aged 17-94 at enrollment (median=53 Q1-Q3=40-70) have been followed for up to 21 visits spanning 40 years . Over 3 days, participants receive comprehensive examinations, interviews, imaging and functional and cognitive evaluations; repeat visits occur every 1-4 years depending on age. The BLSA offers opportunities to examine distributions of, change in and interrelationships among several rarely concurrently ascertained parameters over the life course and across birth cohorts. The BLSA also maintains an extensive biorepository. This talk will summarize the extensive measurement catalogue and timeline and provide illustrative examples from ongoing research on women\u2019s aging and health."} +{"text": "OBJECTIVES/SPECIFIC AIMS: To understand the role of PGF2a and to characterize a novel cyclooxyrgenase (COX)-independent prostaglandin synthesis pathway in epithelial ovarian cancer. METHODS/STUDY POPULATION: We used high grade epithelial ovarian cancer cell line (OVCAR3) as a model to study our pathway. Our main mode of PGF2a detection is through mass spectrometry. RESULTS/ANTICIPATED RESULTS: Our current results suggest the OVCAR3 cells may synthesize PGF2a independently of COX enzymes. We anticipate this novel pathway may be dependent on the TGFb pathway. DISCUSSION/SIGNIFICANCE OF IMPACT: Understanding the role and synthesis pathway of PGF2a may allow us to uncover a novel therapeutic pathway for high grade ovarian cancer."} +{"text": "Pseudomonas putida in rat nicotine self-administration models of addiction.Smoking and tobacco use continue to be the largest preventable causes of death globally. A novel therapeutic approach has recently been proposed: administration of an enzyme that degrades nicotine, the main addictive component of tobacco, minimizing brain exposure and reducing its reinforcing effects. Pre-clinical proof of concept has been previously established through dosing the amine oxidase NicA2 from This paper describes efforts towards optimizing NicA2 for potential therapeutic use: enhancing potency, improving its pharmacokinetic profile, and attenuating immunogenicity. Libraries randomizing residues located in all 22 active site positions of NicA2 were screened. 58 single mutations with 2- to 19-fold enhanced catalytic activity compared to wt at 10\u2009\u03bcM nicotine were identified. A novel nicotine biosensor assay allowed efficient screening of the many primary hits for activity at nicotine concentrations typically found in smokers. 10 mutants with improved activity in rat serum at or below 250\u2009nM were identified. These catalytic improvements translated to increased potency in vivo in the form of further lowering of nicotine blood levels and nicotine accumulation in the brains of Sprague-Dawley rats. Examination of the X-ray crystal structure suggests that these mutants may accelerate the rate limiting re-oxidation of the flavin adenine dinucleotide cofactor by enhancing molecular oxygen\u2019s access. PEGylation of NicA2 led to prolonged serum half-life and lowered immunogenicity observed in a human HLA DR4 transgenic mouse model, without impacting nicotine degrading activity.Systematic mutational analysis of the active site of the nicotine-degrading enzyme NicA2 has yielded 10 variants that increase the catalytic activity and its effects on nicotine distribution in vivo at nicotine plasma concentrations found in smokers. In addition, PEGylation substantially increases circulating half-life and reduces the enzyme\u2019s immunogenic potential. Taken together, these results provide a viable path towards generation of a drug candidate suitable for human therapeutic use in treating nicotine addiction. Smoking is a global healthcare problem . The WorPseudomonas putida in well-accepted rat behavioral models of nicotine addiction \u2009=\u2009kcat/Km*[S]) and to construct progress curves. The progress curves were fit to a simple kinetic model using KinTek software using methodology as described to derive apparent kcat values thymidine addition on day 5.Local Research Ethics Committee approval was granted prior to study commencement. Peripheral blood (60\u2009mL) was obtained from healthy volunteers (escribed . T cell 3H]-Thymidine incorporation in counts per minute (cpm). Analysis was performed by calculating a stimulation index of T cell proliferation by dividing the cpm value obtained for test samples with the baseline cpm value. A fold increase in IFN-\u03b3 levels (pg/mL) was calculated by dividing the value of cells treated with each drug with the baseline value. The cut off value of a 3-fold increase [T cell proliferation was measured by [increase was consEqual numbers of age-matched male and female Sprague Dawley rats (Charles River Laboratories) or HLA transgenic mice per study group were used. Study animals with surgical modifications were housed individually in disposable microisolator cages (Innovive). Environmental controls were set to maintain the following conditions: a temperature range of 64 to 79\u2009\u00b0F, a relative humidity range of 30 to 70%, ten or greater air changes/h, and a 12-h light/12-h dark cycle. Food and water were available ad libitum. Animal welfare followed the NIH guide for Care and Use of Laboratory Animals (8th ed.) and all protocols were approved by Noble Life Sciences (NLS) Institutional Animal Care and Use Committee. NLS is an AAALACi accredited and USDA Licensed (51-12-0092) and OLAW Assured (A4633\u201301) facility. Collection of blood from animals occurred while under isoflurane anesthesia and steps necessary to minimize animal suffering were undertaken. Study animals were euthanized after terminal blood collection by thoracotomy under isoflurane anesthesia consistent with AVMA Guidelines."} +{"text": "Bradyrhizobium sp. strain USDA 3456 is a historic strain from the United States Department of Agriculture (USDA) Agricultural Research Service (ARS) National Rhizobium Germplasm Collection isolated from Vigna unguiculata (cowpea) in 1966. Strain USDA 3456 has been utilized in global agricultural applications, including improving soil nitrogen fertility. The draft genome sequence here provides a genetic reference of a novel diazotroph. Bradyrhizobium sp. strain USDA 3456 is a historic strain from the United States Department of Agriculture (USDA) Agricultural Research Service (ARS) National Rhizobium Germplasm Collection isolated from Vigna unguiculata (cowpea) in 1966. Strain USDA 3456 has been utilized in global agricultural applications, including improving soil nitrogen fertility. The draft genome sequence here provides a genetic reference of a novel diazotroph. The rhizosphere microbiome is one of the most dynamic interfaces on Earth \u20133, with \u2013Bradyrhizobium sp. strain USDA 3456 was isolated from a Vigna unguiculata (cowpea) nodule from Wisconsin in 1966 (Arachis hypogaea) suggest that it possesses high phosphorus solubilization and moderate indole acetic acid (IAA) production (2H4/g\u22121\u2009(nodule dry weight)/h . A single colony was inoculated in AG broth culture (25\u2009ml) at 30\u00b0C at 200 rpm to obtain biomass for DNA extraction (A lyophilized culture of traction .https://seqonce.com/rhinoseq/). Libraries were quantified and sequenced on a HiSeq 4000 instrument in a 150-bp paired-end read format at the Michigan State University Research Technology Support Facility (RTSF).DNA was extracted using the MasterPure DNA extraction kit following the manufacturer\u2019s protocols. A SeqOnce RhinoSeq kit was used to prepare libraries . The resN50 value of 324,457\u2009bp. We estimated completeness and contamination of the draft genome using CheckM (version 1.0.12); the genome was 100% complete with 0% contamination , and code used to generate the assembly can be found at www.github.com/friesenlab/Bradyrhizobium_USDA3456.This whole-genome shotgun project has been deposited at DDBJ/ENA/GenBank under the accession number"} +{"text": "Serine protease high temperature requirement protease A2 (HtrA2) is involved in apoptosis and protein quality control. However, one of its murine inactive mutants (S276C aka mnd2) is associated with motor neuron degeneration 2. Similarly, this conserved mutation in human HtrA2 (hHtrA2) also renders the protease inactive, implicating pathogenicity. However, the structural determinants for its inactivation have not yet been elucidated. Here, using multidisciplinary approach, we studied the structural basis of inactivity associated with this mutation in hHtrA2. Characterization of secondary and tertiary structural properties, protein stability, oligomeric properties, and enzyme activity for both wild-type and mutant has been performed using biophysical and functional enzymology studies. The structural comparison at atomic resolution has been carried out using X-ray crystallography. While enzyme kinetics showed inactivity, spectroscopic probes did not identify any significant secondary structural changes in the mutant. X-ray crystallographic analysis of the mutant protein at 2 \u00c5 resolution highlighted the significance of a water molecule that plays important role in mediating intermolecular interactions for maintaining the functional ensemble of the protease. Overall, the crystallographic data along with biophysical and enzymology studies helped decipher the structural basis of inactivity of hHtrA2S276C, which might pave way toward further investigating its correlation with aberration of normal cellular functions, hence pathogenicity. Escherichia coli that degraded misfolded/unfolded proteins formed during excessive stress conditions or protein\u2013protein interaction domain(s) . This observation was validated by solving structure of hHtrA2S306A under identical crystallization conditions as S276C mutant. It is well established that the contribution of water molecules to a protein\u2019s three dimensional structure is phenomenal. Hydration of protein structure is very important for maintaining its overall tertiary/quaternary structure [Therefore, to understand the structural basis of inactivity in human counterpart (hHtrA2S276C) of mnd2 mouse mutant, high resolution crystal structure was solved followed by elucidation of its biophysical properties. Although, the mutant showed no overall secondary structural changes, conformational stability (Supplem"} +{"text": "This data article presents mean serum concentrations (wet weight and lipid standardized) of 32 persistent organic pollutants (POPs) detected in >75% of participants of the Coronary Artery Risk Development in Young Adults (CARDIA) study across levels of POPs scores, and their corresponding coefficients of determination. POPs scores were calculated as: A) the sum of each participant's log-transformed POPs concentrations as the sum of the participants' log-transformed concentrations of each POP divided by the groups' standard deviation of the corresponding log-transformed POP in early adulthood and blood lipids over a 23-year follow-up\u201d . HDL concentrations were measured via precipitation using dextran sulfate-magnesium chloride on the ABA 200 Biochromatic device. LDL concentrations were calculated using the Friedewald equation 2.3individual/logPOP standard deviationgroup]). We created similar summary variables for 8 organochlorine pesticides (8 OCP summary score), and 23 PCBs (23 PCB summary score); summary variables were created for both wet weight and lipid standardized concentrations.This article presents median POPs concentrations in serum across categories of POPs composite variables. Composite variables are used because the general population is exposed to a mixture of many POPs 2, Pearson correlation) for each association."} +{"text": "Heart failure (HF) is a leading cause of potentially preventable hospital readmissions for Medicare beneficiaries from skilled nursing facilities (SNFs). This research seeks to determine if a HF patient\u2019s insurance type (Medicare Fee-for-Service (FFS) vs. Medicare Advantage (MA)) influences their risk for readmission within 30 days of hospital discharge to a SNF. MA beneficiaries receive benefits through managed care plans with restricted networks, but typically expanded benefits. This research is particularly timely in light of CMS\u2019 new penalties under the Protecting Access to Medicare Act (PAMA) directed at SNFs for 30-day rehospitalizations. Outcomes data on readmissions from a randomized controlled trial of HF Disease Management in SNFs conducted from 2014-2017 were used to evaluate the risk of readmission. Patients with HF receiving SNF care were enrolled and followed for 30 days from SNF admission. Patients were recruited from 29 primarily for-profit (93%) SNFs that contracted with an average of 4.07 (\u00b15.48) MA plans. Of the 340 study participants followed, 62% had FFS Medicare coverage (n=212) and 38% had MA (n=128). In total, 23% (n=79) of patients experienced at least one readmission within 30 days of hospital discharge. FFS patients had a higher risk of rehospitalization within 30 days of hospital discharge than MA patients (25% vs. 20%), but the association between insurance type and rehospitalization was not statistically significant . Findings suggest that insurance type may be an important risk factor for rehospitalizations for patients with HF from SNF; however, a larger sample will need to confirm this relationship."} +{"text": "Pancreatic cancer remains a devastating disease with dismal outcomes despite the development of novel chemotherapeutic regimens and radiation techniques. Stereotactic body radiation therapy (SBRT) offers an advantage both in image guidance and radiation dose delivery to direct ablative doses to tumors with acceptable toxicity compared to conventional techniques. Recent literature is clustered with data pertaining to SBRT in patients with resectable, borderline resectable and locally advanced pancreatic tumors. We here present a summary of the current data and highlight the limitations and potential for future growth. Further clinical study in the form of multi-institutional trials is warranted to establish the role of SBRT in combination with new chemo- therapeutic agents as well as a non-invasive alternative to surgery. Favorable results of SBRT for locally advanced pancreatic cancer (LAPC) patients are now leading to the exploration of SBRT for other pancreatic cancer patients.2Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy with limited effective therapeutic options and exceedingly high mortality. Currently, a cure may be achieved through resection; recent evidence suggests that neoadjuvant therapy can increase R0 resection rates with effective local control.3 The European F\u00e9d\u00e9ration Francophone de Canc\u00e9rologie Digestive (FFCD)/The Soci\u00e9t\u00e9 Francophone de Radioth\u00e9rapie Oncologique (SFRO) Phase III trial compared GEM alone versus induction 5 Fluorouracil (FU) and cisplatin chemoradiation (CRT), followed by maintenance gem.4 Overall survival (OS) was shortened in the CRT arm from 13 to 8.6-months. Higher grade 3 toxicities with CRT were observed during both induction (36% vs. 22%) and maintenance (32% vs. 18%) phases. Notably, the trial utilized a higher than normal conventionally fractionated 60 Gy dose. The recent success of more aggressive, but increasingly toxic, chemotherapy regimens such as FOLFIRINOX and gem plus nab-paclitaxel have spurred re-examination of local therapy.6 With improved systemic control, local progression may become a more serious issue for survival and quality of life. However, local control rates from standard external beam radiotherapy (EBRT) have been disappointing with 1-year local progression rates of around 50%.7 Furthermore, with two-thirds of patients failing distantly within 1 year, a shorter course approach with minimal interruption to systemic therapy is desirable.7 These factors paved the way for the use of SBRT in pancreatic cancer patients, and initially those with LAPC.The utility of pancreatic SBRT was established in the locally advanced patient population. With the advent of gemcitabine-based (GEM) chemotherapy, the role of RT for LAPC has become more precarious.8 The trial was stopped at a dose of 25 Gy since all patients achieved local control with distant metastasis as the first site of failure. The median survival for all patients was 11-months, with 100% local control. However, despite smaller margins and less acute toxicity, patients treated on the Stanford single-fraction SBRT protocol experienced a high degree of late toxicities (25% grade \u22652).9 Hypofractionated studies showed reduced 1-year grade 2 toxicity to 7.8%. This reduction came without a compromise in disease control. The 1-year local control was 91.5% vs. 88.3% (p=0.8) for single vs. 5-fraction SBRT with median OS of 13.6-months for all patients. More contemporary SBRT series have also largely employed a fractionated approach.14 These institutional studies re- veal a median survival of 14\u201315-months, 1-year local control rates of about 80%, and grade 3 toxicities below 10%.15The inception of SBRT for pancreatic cancers began at Stanford with a phase I dose escalation study in a LAPC cohort.16 SBRT doses ranged from 25\u201333 Gy in 5 fractions. The 1-year local control rate was 61%, but with a median OS of 18.4-months for LAPC patients. Notably, 20% of LAPC patients underwent surgery. Resected patients had a median OS of 20.2-months, compared to 12.3-months for unresected cases. Grade 3 toxicity was below 6%. Similar to the study from Hopkins, SBRT data from Moffitt also shows the possibility of downstaging for surgery.14 They reported a 24% surgical conversion rate for LAPC patients receiving FOLFIRINOX chemotherapy. All converted patients achieved an R0 (microscopic negative margin) resection. Any grade 3 or higher toxicity was 7%. Median OS was 34.2-months for patients who underwent resection, and 11.3-months for those who did not. See Very recently, a few groups have reported that LAPC patients may have an increased likelihood of undergoing resection after aggressive induction chemotherapy regimens. Recently, the group from Hopkins reported on 88 patients treated from 2010\u201314 with SBRT using gem-based or FOLFIRINOX regimens.12 SBRT was delivered using 5 consecutive daily fractions targeting the primary tumor with a median dose of 30 Gy , the region of vasculature involvement was prescribed a median dose of 35 Gy using a simultaneous integrated boost (SIB) to further increase the likelihood of tumor regression and R0 resection. After restaging, 56.1% of the BRPC patients underwent surgical resection with all except for one (96.9%) having negative margins. Resected patients had significantly improved median OS and median progression-free survival (PFS) . No acute grade 3 toxicities were reported and the most common acute toxicities were grade 1\u20132 fatigue and nausea. Their subsequent study of 159 patients , surgical resection was performed on 51% of the BRPC patients and R0 resection was achieved in 96%. Portal vein (PV) or superior mesenteric vein (SMV) resection and reconstruction was performed in 34% of BRPC patients. Median OS was significantly higher among patients who had surgery compared to those who did not . Finally, while the prescription doses generally increased compared to the previous publication , the incidence of late grade 3 radiation-related toxicity remained consistently low (~5%).14While pancreatic SBRT has been most extensively evaluated in LAPC patients, there is emerging data that SBRT may also benefit patients with borderline resectable pancreas (BRPC) . The SBR17The feasibility of using SBRT for BRPC is also supported by other studies with more limited numbers of BRPC patients. A study from Johns Hopkins included 88 patients who received 5-fraction SBRT and reported favorable surgical and SBRT-related toxicity outcomes.18 A high rate of R0 resection was achieved (92%) with minimal toxicity. Pathologic complete response (pCR) was achieved in 25%, which is higher than would be expected with standard EBRT and perhaps signaling that SBRT may have unique histopathologic effects. It is plausible that a higher rate of pCR may be achieved using dose fractionation schedules with a higher biologically effective dose. He et al compared surgical outcomes among BRPC/ LAPC patients who received SBRT (n=29), CRT (n=82), or chemotherapy alone (n=26) and reported R0 resection rates of 90%, 84% and 62%, respectively (p=0.02).19 The PCR rate was notably higher among patients who received SBRT .Investigators from the University of Pittsburgh published their experience of 12 patients who received chemotherapy followed by SBRT prescribed to 36 Gy in 3 fractions (n=7) or 24 Gy in a single fraction (n=5) and then had surgery.In conclusion, while various neoadjuvant treatment regimens are commonly used for BRPC including standard fractionation CRT, increasing consideration should be given to SBRT based on its clear advantage in increasing R0 resectability with higher PCR rates, and providing improved OS in these patients.20 Recent studies based on rigorous pathological examination protocols report R1 rates of well over 70%.25 Several studies have shown that residual cancer cells are frequently present in the resection bed even in appropriately staged patients after surgery that is properly performed,26 where even with R0 resections nearly 80% of patients were found to have evidence of microscopic cells left in situ at the surgical site.27 In a recent phase III adjuvant chemotherapy trial in patients with resected pancreatic cancer in which many patients had positive margins (0\u201360%) and nodal involvement (63\u201380%), local recurrence rates were 18\u201341%, suggesting the presence of residual disease may benefit from local therapy in addition to systemic therapy.28 Early data from MD Anderson Cancer Center included 86 patients who received gemcitabine-based X-ray telescope (XRT) radiation (30 Gy); 75% of patients were resected, 95% had R0 resections and the median OS for those who completed all therapy was 34-months.29 Their subsequent study of cisplatin and gemcitabine followed by gemcitabine-based chemoradiation in 90 patients with remote procedure call (RPC) revealed an R0 resection rate of 96% and median OS of 31-months.30 Cloyd et al published a unique retrospective study utilizing propensity score weighted methodologies. The authors queried MD Anderson database to identify all patients who received pre-operative chemotherapy or CRT before pancreatectomy for anatomically resectable PDAC between 1999 and 2014. They concluded that the receipt of pre-operative CRT alone was associated with a higher rate of margin-negative resection , lower rate of positive lymph nodes , greater treatment effect, reduced incidence of locoregional recurrence (LR) but similar median overall survival compared with systemic chemotherapy alone.31 Katz et al, report- ed wider special memorandum account (SMA) margin distance on histological examination on patients who receive pre-operative CRT.32 This suggests that the local effect of CRT may occur primarily through sterilization of the retroperitoneum.The significance of microscopic margin involvement on survival is a controversial topic, with some studies claiming an impact on survival and others finding no such correlation.37 Damage to tumor cell deoxyribonucleic acid (DNA) is thought to account for only part of the efficacy of hypofractionated regimens.38 Many studies indicate that in addition to the direct impact on DNA, the effects of high-dose radiation on the tumor microenvironment (TME) may play a role in tumor control by SBRT and stereotactic radiosurgery (SRS).41 Many studies indicate the effect of a single fraction or hypofractionated radiation therapy in the treatment of pancreatic tumor xenografts.Both SBRT and SRS have been used effectively for the treatment of lung, liver, brain, prostate, and recurrent head and neck cancers, among others.46 CAFs are also responsible for the deposition of key extracellular matrix (ECM) proteins as well as secreting ECM-degrading enzymes ,43 which promotes migration of CAFs and degradation of the ECM, allowing the invasion of tumor cells.47In the stroma of human carcinomas, cancer-associated fibroblasts (CAFs) are the most abundant cell types and play a significant role in tumor cell growth, angiogenesis, and invasiveness .42\u201346 CAIn vitro studies have shown that fibroblasts develop an irreversible senescent phenotype when exposed to a dose>10 Gy of radiation, whereas low doses of radiation induce reversible DNA damage without growth arrest. Senescent fibroblasts release proteolytic enzymes, cytokines, growth factors, and reactive oxygen species, creating a protumorigenic environment.48 Radiation doses higher than 10 Gy per fraction are associated with severe vascular damage leading to the deterioration of the TME.49 Although endothelial cell damage has been shown to be a major factor in the biological mechanism of SBRT and SRS, this phenomenon is sometimes transient and may lead to neovasculogenesis via hypoxia-inducible factor (HIF)-1 induction.49 Baird et al reported pancreatic tumor regression through activation of type 1 interferon-dependent responses with a single dose of 10 Gy and co-treatment with cGAMP or STING (simulator of interferon genes) agonists that amplify the radiation-induced antitumor immune response.51 Type 1 interferons (interferon (IFN)-\u03b1 and IFN-\u03b2) are important for activation of both innate and adaptive immune responses and are well-known for their role in viral immunity.5253 Likewise, increased infiltration of T-cells into tumors and tumor killing mediated by iNOS+M1 macro- phages through the expression of Type 1 T helper (TH1) cytokines have been reported in murine models of pancreatic cancer and melanoma after low-dose radiation treatment.54 Moreover, many studies have demonstrated M2 polarization after treatment with single high-dose and hypofractionated radiation regimens.57 Several clinical trials are underway to determine the effects of combination therapy with radiation and immune checkpoint inhibitors (60Treatment of pancreatic tumor xenografts with radiation given as 4 Gy in 2 fractions resulted in a switchin tumor-infiltrating macrophages from a protumorigenic M2 phenotype to an antitumorigenic M1 phenotype.hibitors .58\u201360SBRT has been shown to be safe and effective in pancreatic cancer patients. It offers several advantages over standard EBRT including increased patient convenience, reduced toxicities, and the ability to minimize delays in modern multi-agent chemotherapy. The ability of SBRT to convert patients with borderline and locally advanced tumors to resectable disease with higher percentage of negative resection margins may improve survival. Favorable SBRT outcomes for LAPC patients have paved the way for exploration of SBRT for resectable pancreatic cancer patients, with promising early results. The immunotherapeutic approach has very limited clinical activity to date in pancreatic cancer, it is still unclear how to optimally combine ablative radiation and immunotherapy, including optimal sequencing, radiation dose to effectively overcome the immunosuppressive pancreatic tumor microenvironment."} +{"text": "Lung cancer is the leading cause of cancer death worldwide. Cigarette smoking is the most common risk factor for lung carcinoma; other risks include genetic factors and exposure to radon gas, asbestos, secondhand smoke, and air pollution. Nicotine, the primary addictive constituent of cigarettes, contributes to cancer progression through activation of nicotinic acetylcholine receptors (nAChRs), which are membrane ligand-gated ion channels. Activation of nicotine/nAChR signaling is associated with lung cancer risk and drug resistance. We focused on nAChR pathways activated by nicotine and its downstream signaling involved in regulating apoptotic factors of mitochondria and drug resistance in lung cancer. Increasing evidence suggests that several sirtuins play a critical role in multiple aspects of cancer drug resistance. Thus, understanding the consequences of crosstalk between nicotine/nAChRs and sirtuin signaling pathways in the regulation of drug resistance could be a critical implication for cancer therapy. Globally, lung cancer is greatest cause of cancer-related deaths. Lung cancer accounts for 14% and 12% of all cancers in men and women, respectively, and represents 24.6% of all cancer-related deaths a]pyrene (BaP), polycyclic aromatic hydrocarbons (PAH), nicotine, and nitrosamines Chemicals in cigarette smoke enter the bloodstream and affect the body; thus, smoking causes many diseases including cardiovascular disease, chronic obstructive pulmonary disease (COPD), and lung cancer In addition to nicotine, its oncogenic derivatives 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N-nitrosonornicotine (NNN), present in tobacco smoke, can activate nAChRs signaling and stimulate multiple cancer-promoting signaling Cigarette smoke is associated with an increased risk of all histological types of lung cancer Survival analysis of a Cancer Genome Atlas (TCGA) lung cancer dataset demonstrated that high expression of acetylcholine receptors (AChRs) gene family such as CHRM2, CHRM3, CHRNA1, CHRNA2, CHRNA6, CHRNB3, or CHRNE is associated with favorable prognosis in NSCLC adenocarcinoma, but that of CHRNA5/\u03b15nAChR or CHRNA7/\u03b17nAChR is associated with an unfavorable prognosis Single nucleotide polymorphisms (SNPs) located on chromosome 15q25, which contains the nAChR subunits encoding by the CHRNA5, CHRNA3, and CHRNB4, are associated with lung cancer risk Nicotine may induce \u03b17nAChR expression in human SCLC cells via the Sp1/GATA regulation signaling pathway 2+\u03b17nAChR mediates the proliferative effects of nicotine in lung cancer cells Cigarette smoke status and history are associated with lung cancer metastasis \u03b17nAChR enhances angiogenesis via the PI3K/AKT pathway and NF-\u03baB activation, which is partially dependent on vascular endothelial growth factor (VEGF) \u03b17nAChR attenuates ventilator-induced lung injury and plays an anti-inflammatory role in several inflammatory diseases Exposure to nicotine adversely affects dendritic cells, a cell type that has an important role in anticancer immunosurveillance Sirtuins play diverse roles in controlling the cell cycle and proliferation in response to stress, thus promoting cell survival, apoptosis, or senescence Sirtuins are responsible for cellular metabolic reprogramming and drug resistance by inactivating cell death pathways and promoting uncontrolled proliferation The SIRT1 inhibitor sirtinol induces senescence-like growth arrest through impaired activation of RAS-mitogen-activated protein kinase (MAPK) signaling in human lung cancer cells Nicotine may reduce the cytotoxic effects of chemotherapy and radiotherapy that cause poor therapeutic response Nicotine also permeated cells and activated mitochondrion-nAChRs coupling to inhibit mitochondrial permeability transition pore (mPTP) opening, preventing apoptosis +-dependent deacetylases and are implicated in the oxidative stress response through the regulation of mitochondrial metabolism and antioxidant mechanisms Sirtuins can exert their capacity to respond to environmental changes and their expression is often altered in cancer Nicotine can upregulate SIRT1 expression in a time- and concentration-dependent manner 2O2 or etoposide treatment SIRT3, located in mitochondria, is correlated with NSCLC malignancy in vivo (mice) and in vitro Patients with high cytosol expression but low nuclear expression of SIRT6 can have poor clinical outcomes of lung cancer Thus, the aforementioned studies together have demonstrated tumor progression modulated by the SIRT1, SIRT3, and SIRT5-7, along with the tumor-suppressive effects of SIRT2 and SIRT4. SIRT2 mediates the ROS production and p27 levels, leading to lung cancer cell apoptosis and cell-cycle arrest Cell-membrane nAChRs implement upregulation of proliferative and survival genes +/SIRT1 pathway, which promotes chemotherapeutic drug resistance Genome-wide association studies have indicated a strong link between nicotine/nAChRs and lung cancer risk Nicotine activates \u03b17nAChR and \u03b22nAChR, and these two factors are associated with EGF and VEGEF receptors, respectively. However, activated mitochondrial \u03b17nAChRs and \u03b24nAChR interacted with the PI3K and Src An accumulating body of evidence is demonstrating that long noncoding RNAs (lncRNAs) have various biological functions, including modulation of growth, cell differentiation, drug resistance, and cancer progression Over the past few years, nAChRs have been found to be selectively overexpressed in various cancers, including lung cancer. Targeting nAChRs signaling pathways can significantly attenuate nicotine-associated drug resistance. Several \u03b17nAChR antagonists are potential anticancer drugs Nicotine and derived metabolites are associated with lung cancer risk in smokers. The \u03b17nAChR is significantly upregulated in NSCLC and correlates with its unfavorable prognosis. Activation of nicotine/\u03b17nAChR signaling leads to lung cancer progression. Studies have suggested that nicotine/nAChR axis and SIRT1/3/5-7 mediates cancer drug resistance. Blockade of the signaling mediated by nicotine/nAChR and specific sirtuins may enhance the efficacy of chemotherapy."} +{"text": "E. coli) measurements to elucidate water quality fecal pollution trends. Results demonstrate the feasibility of implementing standardized fecal source identification qPCR methods with established data acceptance metrics in a large-scale field study leading to new investigative leads suggesting that elevated E. coli levels may be linked to specific pollution sources and land use activities in the Tillamook Bay Watershed.Fecal pollution management remains one of the biggest challenges for water quality authorities worldwide. Advanced fecal pollution source identification technologies are now available that can provide quantitative information from many animal groups. As public interest in these methodologies grows, it is vital to use standardized procedures with clearly defined data acceptance metrics and conduct field studies demonstrating the use of these techniques to help resolve real-world water quality challenges. Here we apply recently standardized human-associated qPCR methods with custom data acceptance metrics (HF183/BacR287 and HumM2), along with established procedures for ruminant (Rum2Bac), cattle (CowM2 and CowM3), canine (DG3 and DG37), and avian (GFD) fecal pollution sources to (i) demonstrate the feasibility of implementing standardized qPCR procedures in a large-scale field study, and (ii) characterize trends in fecal pollution sources in the research area. A total of 602 water samples were collected over a one-year period at 29 sites along the Trask, Kilchis, and Tillamook rivers and tributaries in the Tillamook Bay Watershed . Host-associated qPCR results were combined with high-resolution geographic information system (GIS) land use and general indicator bacteria ( E. coli or enterococci for routine monitoring to identify locations with unsafe levels of fecal pollution and amplification efficiency (0.90 to 1.10 where E = 10(-1/slope)\u2013 1) acceptance criteria, as well as instrument run-specific IAC proficiency testing .A series of acceptance metrics designed to ensure high quality data generation were used in this study . Briefly10 copy number per reaction. Sensitivity was defined as the total number of correct positive reactions divided by the total number of reactions containing the target pollution source (sensitivity = TP/(FN + TP), where TP and FN are true positives and false negatives, respectively). Specificity was calculated as the total number of correctly identified negative reactions divided the total number of reaction that do not contain the target pollution source (specificity = TN/(FP + TN) where TN and FP are true negatives and false positives, respectively). To investigate potential land use and weather trends in qPCR measurements, average log10 copies per reaction were estimated using a maximum likelihood estimation method by either sampling site (land use) or sampling day (weather). Average log10 MPN/100mL values were used for E. coli measurements. Water quality metrics were considered eligible for trend analysis if more than 20% of respective average concentrations (sampling site or sampling day) were greater than zero. For weather trend analysis, eligible sampling day log10 copies per reaction concentrations were binned into two groups based on paired precipitation, solar irradiance, or air temperature median values . Sample sites were ranked for each qPCR assay using a weighted average fecal score utilizing all measurements including non-detects as reported elsewhere [Master calibration models were generated for each qPCR assay from six independent standard curves using a Bayesian Markov Chain Monte Carlo approach . The lown values and subjlsewhere . R stati2 values were all greater than 0.981, and E values ranged from 0.90 (GFD) to 0.97 (HF183/BacR287). Of the 598 water filters, six DNA extracts (1.0%) were discarded from the study due to severe matrix interference based on SPC tests. SPC acceptance thresholds ranged from 21.9 Cq to 25.5 Cq. A total of 217 filter DNA extracts were eligible for Cq adjustments ranging from 0.004 to 3.09. Acceptable DNA recovery was monitored for each extraction batch (n = 38 samples/batch) using the SPC proficiency test [q to 37.8 Cq (HF183/BacR2876) and 34.4 Cq to 38.6 Cq (HumM2). Competition thresholds were 27.9 Cq for HF183/BacR287 and 30.1 Cq for HumM2. Extraneous DNA control reactions indicated 99.95% DNA-free . False positives were both from HF183/BacR287 tests .All qPCR experiments were subject to a rigorous series of quality controls and data acceptance metrics to ensure the use of high-quality data for fecal source identification. Calibration model performance parameters and IAC thresholds for each qPCR assay are shown in The standardization of fecal source identification qPCR laboratory practices and development of data acceptance criteria are critical for these technologies to transition from a subject of environmental microbiology research to useful water quality management and safety planning tools. In 2016, a team of researchers published the first standardized qPCR methodology including custom data acceptance metrics for two top performing human-associated fecal source identification technologies , which wSystematic surveillance of standardized HF183/BacR287 and HumM2 quality control and data acceptance data from the Tillamook Bay Watershed field study revealed two important observations. First, it is feasible to employ all proposed data acceptance metrics in a large-scale study. Notably, proficiency tests specifically designed to ensure proper implementation of DNA recovery (SPC) and amplification inhibition (IAC) controls demonstrated acceptable performance in 92% (DNA recovery) and 100% (amplification inhibition) of experiments. Second, poor DNA recovery and amplification inhibition were absent in more than 98% of water samples tested suggesting that custom environmental reagents and standardized DNA purification practices can consistently yield suitable DNA for genetic testing in environmental conditions. It is important to note that this case study focused on freshwater samples collected from rivers in the Tillamook Bay Watershed. Future large-scale field research studies are warranted to assess the performance of these technologies across a broader range of geographic locations and water types (i.e. marine).E. coli cultivation measurements to characterize fecal pollution in water samples collected from the Tillamook Bay Watershed. Estimated mean log10 copies per reaction concentrations for host-associated qPCR methods are shown in E. coli site average log10 MPN/100mL concentrations ranged from 1.38 (K3) to 2.76 (TR11) (E. coli log10 MPN/100mL concentrations and corresponding rankings for each host-associated genetic marker (average log10 copies/reaction). For a complete list of site rankings, refer to Fecal source identification qPCR methods for human (HF183/BacR287 and HumM2), ruminant (Rum2Bac), cattle (CowM2 and CowM3), canine (DG3 and DG37), and avian (GFD) were combined with 6 (TR11) . Table 2E. coli (100% of sampling site averages > 0), Rum2Bac (75.9%), GFD (34.5%), and HF183/BacR287 (24.1%). Descriptive statistics and daily measurements for air temperature (\u00b0C), solar irradiance (kW\u2027hr/m2), and precipitation (mm) are reported in 2, and no precipitation (per a 72-h accumulation period) was observed on 50% of days (11 of 22). Three water quality metrics were eligible for comparing potential links between water quality measurements and weather parameters including E. coli (100% of sampling day averages > 0), Rum2Bac (55.9%), and GFD (23.8%). Detailed findings are presented and discussed below organized by E. coli, ruminant, avian, human, and canine fecal pollution trends.Water quality measurements were compared to land use and weather data to uncover potential trends in fecal pollution. GIS mapping was used to define catchment drainage boundaries allowing for the estimate of human population , non-sewer (% of catchment area), cropland (% of catchment area), and maximum permitted CAFO cattle population associated with each sampling site . Across E. coli count \u2265 406 MPN/100mL), study sites were impaired from 0% to 80% (TR11) of the time further supporting a link between local weather and the occurrence of fecal pollution. In addition, a significant positive correlation between E. coli and percent non-sewered area was observed suggesting that local septic system use may be a contributing factor. While E. coli testing confirms the presence of unsafe levels of fecal pollution at several sites and suggests rainfall and septic systems are contributing factors to poor water quality in the Tillamook Bay Watershed, these measurements do not specify pollution sources making it difficult to plan cost-effective remediation efforts.Fecal pollution management in the Tillamook Bay Watershed is a year-round challenge. Using the local regulatory criteria for a single grab surface water sample (the time . This brCervus canadensis) and cattle (Bos taurus), CowM2 and CowM3 genetic markers do and can therefore confirm the presence of cattle fecal pollution in a water sample. These methods identified the presence of measurable concentrations of cattle fecal pollution in 8.9% of samples successfully identifying this pollution source at 58.6% (17 of 29) of study sites (10 2.12 copies/reaction). This location also had the highest average concentration of cow-associated CowM2 (log10 0.35 copies/reaction) and CowM3 (log10 1.02 copies/reaction) genetic markers affirming the presence of cattle fecal pollution at this site. The TR3 catchment area includes three dairy cattle CAFO facilities permitted to house up to 2,205 individual animals. Across all study sites, Rum2Bac genetic marker concentrations were positively associated with the permitted number of CAFO cattle and precipitation . In addition, Rum2Bac concentrations were significantly correlated all weather conditions (p \u2264 0.002) suggesting a catchment area source loading potential consistent with rainfall run-off models, in which ruminant pollution builds up on the landscape between storm events and is washed off during subsequent rain storms. In addition, cattle roughly outnumber elk 20:1 in the study area and increases in Rum2Bac genetic marker concentrations were significantly correlated with the number of permitted CAFO cattle. However, it is still possible that elk also contribute to water quality challenges in the Tillamook Bay Watershed. To help identify sampling sites with a potentially high likelihood of elk fecal pollution impact, it will be necessary to obtain more accurate elk population and seasonal movement information or develop an elk-associated genetic marker for future water quality testing.The Tillamook Bay Watershed is home to a large dairy cattle population producing more than 300,000 tons of manure each year . The rumE. coli, as well as a range of pathogens that can potentially infect humans and contribute to poor water quality [Fecal waste from bird species can harbor general fecal indicator bacteria such as quality \u201341. The E. coli, these human waste sources can harbor numerous pathogens .There are an estimated 29,758 residents in the study area, roughly half the number of permitted CAFO cattle. Human waste can potentially enter local waters in the Tillamook Bay Watershed from wastewater treatment plants, public campgrounds, potential stormwater sewer cross-connections, faulty onsite septic systems, seasonal portable restrooms at local parks, and transient camps. In addition to containing poridium ), solidsporidium ). The inporidium , no measE. coli [E. coli 80% of the time to 100% suggesting that if a host-associated genetic marker was observed in a water sample, the presence of the respective pollution source can be interpreted with a high degree of confidence. Second, Rum2Bac, CowM2, and CowM3 genetic markers were undetectable in all calf samples (< 115 days in age), but present in 36.5% (CowM2), 82.3% (CowM3), and 93% (Rum2Bac) of adult samples tested (> 6 months in age) indicating that these methods could underestimate the total cattle impact (juvenile + adults) in the study area. This same trend was reported in a dairy calf population from another agricultural facility suggesting these ruminant-associated genetic markers may be absence in calves across the United States . Third, Other notable observations from reference pollution source testing include a higher prevalence of CowM3 compared to CowM2, likely due to local feeding practices ,50. A loMeasurements whose values are known only to be above or below a defined threshold are called \u2018censored\u2019 data. For qPCR fecal source identification applications, a censored data situation occurs when samples yield Cq values greater than the respective LLOQ threshold . This creates a censor data challenge in the sense that the true number of DNA target molecules in the sample cannot be firmly established. For fecal source identification qPCR data interpretation, these values are often deleted ,52 or thThe objectives of this study were to implement the HF183/BacR287 and HumM2 human-associated qPCR standardized laboratory and data acceptance procedures in a large-scale field study and elucidate fecal pollution dynamics in host-associated genetic markers measured by qPCR across the Tillamook Bay Watershed. Intensive fecal source characterization of more than 600 samples collected from 29 sites over a year clearly demonstrates the feasibility and value of using standardized laboratory procedures and data acceptance metrics, especially for the HF183/BacR287 and HumM2 human-associated qPCR methods. Host-associated qPCR testing successfully uncovered numerous fecal pollution trends in the Tillamook Bay Watershed offering a multitude of new information to help local authorities improve water quality management. In addition, this large-scale field demonstration revealed key issues regarding qPCR data interpretation such as the importance of confirming method performance with local reference fecal samples and utilizing appropriate censored data analysis strategies. Further investigation of this rich data set will likely lead to additional water quality management information. Finally, it will be vital to continue to conduct large-scale, intensive field studies, such as the study presented here, in other water quality management arenas such as urban stormwater scenarios and recreational beach settings to tailor qPCR data interpretation strategies for these different applications.S1 Fig(TIF)Click here for additional data file.S2 Fig2; middle), and 120-h precipitation are shown. Black diamonds indicate sampling event time points.Air temperature , solar irradiance (kW\u2027hr/m(TIF)Click here for additional data file.S3 Fig(TIF)Click here for additional data file.S1 TableE. coli exceedance across study area.Sampling site information and historical trends in single-day maximum (PDF)Click here for additional data file.S2 Table(PDF)Click here for additional data file.S3 Table(PDF)Click here for additional data file.S4 Table(PDF)Click here for additional data file.S5 Table(PDF)Click here for additional data file."} +{"text": "Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Trichomonas vaginalis (TV) as well as human papillomavirus (HPV), and persistent cervical inflammation is one of the etiologic agents of cervical cancer. Toll-like receptors (TLRs) play an important role in the recognition and subsequent elimination of these pathogens. Variations in the Toll-like receptor genes influence susceptibility to pathogens as well as disease progression independently.Cervicitis is one of the major health problems amongst women caused by infection of various pathogens including TLR4 and TLR9 genes were analyzed among 130 cervicitis patients and 150 controls either using polymerase chain reaction-restriction fragment length polymorphism or allele specific-PCR.Ten single nucleotide polymorphisms, five each of T. vaginalis infection was found at the highest frequency (30.7%) as compared to C. trachomatis (1.5%), N. gonorrhoeae (2.3%) and HPV (4.6%) infections in cervicitis patients. TLR4 rs11536889 CC and TLR9 rs187084 TC genotypes showed the higher distribution in cervicitis patients compared to controls. In addition, TLR4 rs11536889 C allele was shown to increase the risk of cervicitis compared to controls. The TLR4 haplotype GCA and TLR9 haplotype GTA were found to be associated with decreased and increased risk of cervicitis respectively.TLR4 and TLR9 polymorphisms, as well as haplotypes were shown to modulate the cervicitis risk. Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG). Infections of Trichomonas vaginalis (TV) and human papillomavirus (HPV) have also been implicated in the pathogenesis of cervicitis, in addition to other bacterial and viral pathogens ) , Iran or intronic rs1927911 polymorphism to be associated with cervicitis. The Asp299Gly change was associated with inflammatory bowel diseases [TLR4 intronic SNP, rs1927911 has been reported to increase the risk of diabetic foot ulcers [On the other hand, we did not find either diseases . In the t ulcers and athet ulcers .TLR9 gene polymorphisms, we found promoter rs187084 TC genotype to be associated with an increased risk of cervicitis. A similar result was obtained in inflammatory bowel diseases [TLR9 rs5743844, rs352140, rs5743836 or rs352139 polymorphisms were associated with cervicitis. A complete absence of TLR9 non-synonymous polymorphism rs5743844 (Pro99Leu) in our study population corroborates with the report of Lee and group (2006) where neither controls nor lung disease patients carried the same polymorphism [With regard to diseases . On the morphism . Howevermorphism . The disTLR4 and TLR9 SNPs. TLR4 GCA and TLR9 GTA haplotypes were significantly associated with decreased and increased risk of cervicitis respectively. Moreover, within cervicitis cases, haplotype AGC was found to be significantly associated with TV induced cervicitis. Our results indicate that two SNPs each in both TLR4 , and TLR9 genes were in strong LD. Furthermore, certain SNP pairs in our study deviated from the norm that the linkage disequilibrium is a function of distance, which is accordance with the observations of Stephens et al. (2001) [TLR4 SNPs (rs10759931 and rs4986790) that were separated by a distance of 11.1 kb showed strong LD (D\u2032 = 0.62) while the SNPs (rs4986790 and rs11536889) that were separated by a shorter 2.8 kb distance did not exhibit a strong linkage disequilibrium (D\u2032 = 0.04). The SNP pairs rs352140:rs187084 and rs5743836:rs187084 of TLR9 gene were also in agreement with the above-mentioned trend, where the SNP pair rs352140:rs187084 that were separated by a larger distance (4.3kb) exhibited a stronger LD (D\u2032 = 0.52) as compared to the pair (rs5743836:rs187084) that were separated by a comparatively smaller distance . TLR4 and TLR9 gene, the genetic distance between SNPs and the D\u02c8 values.As haplotypes are considered more informative than SNPs , we gene. (2001) . The TLRTLR polymorphisms, we found TLR4 rs11536889 CC genotype to be significantly associated with higher risk of TV induced cervicitis. None of the other TLR4 and TLR9 SNPs or haplotypes showed association with TV infection. No research group has yet investigated the role of TLR polymorphisms in TV induced cervicitis. However, Chen et al., (2013) observed a marginal association of the TLR4 rs4986790 AG genotype with TV infected prostate cancer patients. In the case of CT, NG, and HPV infections and their association with TLR SNPs, limited reports are available worldwide, and none is available on cervicitis. Several reports demonstrate that TLR4 and TLR9 polymorphism are associated with CT and NG (TLR4: rs1927911 and rs4986790 with PID) infections and disease association [TLR9 promoter rs5743836 polymorphism with HPV clearance or persistence healthy women [TLR4 or TLR9 SNPs and haplotypes. Moreover, exploring the effect of above said polymorphisms on the expression pattern of TLR4 and TLR9 genes could provide more insights on the influence of CT, NG, TV, HPV infections on cytokine production and the host immune response.Coming back to pathogen infection and ociation \u201329. On thy women . Due to TLR4 and TLR9 polymorphisms on cervicitis. However, our study also suffered from many limitations. For example, being a hospital-based case-control study, the selection bias could not be excluded. Study on the expression pattern of the TLR4 and TLR9 would have reflected the effect of SNPs. Last but not the least, HPV16 and 18 copy number analysis could have also revealed a probable link between TLR4 and TLR9 polymorphisms and their effect of severity of HPV infection.Based on our results, we suggest a significant influence of TLR4 and TLR9 SNPs as well as haplotypes modulated the cervicitis risk as a whole and TV induced cervicitis as well. Furthermore, elucidation of the functional role of these polymorphisms may help in understanding the pathophysiology of cervicitis. Our results provide lead-in information to develop personalized clinical marker that could be utilised in future as a screening tool. This may be useful in providing primary preventive care by identifying women at greater risk of cervicitis and possibly cervical cancer. Finally, a comprehensive multicentric study on large and varied ethnic populations will help in precisely understanding the clinical relevance and overall impact of both the genes to CT, NG, TV, HPV infections and cervicitis risk.Cervicitis is though curable using antibiotic regime , it is wS1 Figa and d represents haplotype block structures generated excluding rs10759931 and rs11536889 respectively. b and e shows linkage disequilibrium plots generated excluding rs10759931 and rs11536889 respectively, representing the degree of linkage disequilibrium between two SNPs, indicated by the level of pair-wise D\u2019 values shown in the blocks. c and f represents the r2 values generated excluding rs10759931 with percentage correlation between the two SNPs shown in each box.(TIF)Click here for additional data file.S2 Figa and d represents haplotype block structures generated excluding rs187084 and rs5743836; respectively. b and e shows linkage disequilibrium plots generated excluding rs187084 and rs5743836 respectively, representing the degree of linkage disequilibrium between two SNPs, indicated by the level of pair-wise D\u2019 values shown in the blocks. c and f represents the r2 values generated excluding rs187084 and rs5743836 respectively, with percentage correlation between the two SNPs shown in each box.(TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.S4 Table(DOCX)Click here for additional data file.S5 Table(DOCX)Click here for additional data file.S6 Table(DOCX)Click here for additional data file.S7 Table(DOCX)Click here for additional data file.S8 Table(DOCX)Click here for additional data file.S9 Table(DOCX)Click here for additional data file.S10 Table(DOCX)Click here for additional data file.S1 Dataset(XLSX)Click here for additional data file."} +{"text": "Escherichia coli (E. coli) in porcine mammary epithelial cells (PMEC). Two time points (3\u2009h and 24\u2009h) were selected based on specific transcriptomic changes during the early and late immune responses, respectively.Epigenetic changes such as cytosine (CpG) DNA methylations regulate gene expression patterns in response to environmental cues including infections. Microbial infections induce DNA methylations that play a potential role in modulating host-immune response. In the present study, we sought to determine DNA methylation changes induced by the mastitis causing P\u2009<\u00a00.01) differentially methylated CpG sites at 3\u2009h and 24\u2009h after E. coli challenge in PMEC respectively. These CpG sites mapped to genes that have functional roles in innate and adaptive immune responses. Significantly, hypomethylated CpG sites were found in the promoter regions of immune response genes such as SDF4, SRXN1, CSF1 and CXCL14. The quantitative transcript estimation indicated higher expression associated with the DNA CpG methylation observed in these immune response genes. Further, E. coli challenge significantly reduced the expression levels of DNMT3a, a subtype of de novo DNA methylation enzyme, in PMEC indicating the probable reason for the hypomethylation observed in the immune response genes.DNA methylation analysis revealed 561 and 898 significant contains supplementary material, which is available to authorized users. DNMT1) [Escherichia coli infection (UPEC) of human uroepithelial cells was reported to regulate the expression of DNMT1, and CpG hypermethylation down-regulated the cell-cycle inhibitor CDKN2A likely inhibiting apoptosis and increasing proliferation of uroepithelial cells [Leishmania donovani also alters DNA methylation profiles in human macrophages suppressing the host immune response and enabling intracellular survival of the protozoan [Mycobacterium tuberculosis (TB) infection that rapidly methylates host DNA at distal enhancer elements and associated chromatin remodelling [Epigenomic changes are dynamically regulated by environmental cues. Among the various epigenetic modifications, cytosine (CpG) methylation of genomic DNA is an important reversible gene regulatory mechanism. DNA methylation plays a crucial role in transcriptional regulation by affecting the recruitment of regulatory factors onto promoters and enhancers . MicrobiDNMT1) . Escherial cells . Infectirotozoan . Similarodelling . Schistoodelling . These eEscherichia coli (E. coli) are the most prominent causative pathogens isolated from PDS- affected sows. Lipopolysaccharide (LPS), an outer membrane endotoxin component of E. coli, is the major pathogenic factor that can induce inflammatory responses in sows with PDS [E. coli bacterial challenge of porcine mammary epithelial cells (PMECs) was used as a model for porcine mastitis at two time points (3\u2009h and 24\u2009h) that represented early and late transcriptional responses [E. coli in porcine mammary epithelial cells and found upregulated set of genes, including of cytokines, chemokines, and cell adhesion factors, which together coordinate the immune response of host cells [E. coli induced transcriptomic changes in PMEC also follow other epigenomic modifications. Therefore, our present study focuses on host-cell DNA methylation changes induced by the mastitis causing E. coli.Coliform mastitis (CM) causes postpartum dysagalactia syndrome (PDS), an important disease in pigs. The affected animals show high fever, loss of appetite, pain and inflammation of teats. PDS is a disease of economic significance as it severely affects the health and milk production of sows leading to poor survival of piglets . Gram-newith PDS . In our esponses . There wst cells . These tst cells . We hypoE. coli and 24hpc E. coli) generated 25\u201340 million Illumina sequencing reads for each of the nine reduced-representation bisulfite sequencing (RRBS) libraries. Mapping to the pig genome (Sscrofa 11.1) using Bismark (Bowtie 2) revealed that overall 50% of the generated reads uniquely mapped to the genome. Twenty percent of the CpGs analysed across all samples mapped to the 5\u2032 regulatory region (promoter) of genes with functional annotations. Further, approximately 45% of mapped CpGs were enriched in known CpG islands of the pig genome. The average CpG methylation levels was approximately 45% and the non-CpG level was less than 10% in both the control and treated groups Sscrofa 1.1 usingE. coli 3 hpc group, between control and E. coli 24\u2009hpc group and between E. coli 3 hpc group and 24\u2009hpc group, respectively, were further used for differential methylation analysis. There were a total of 561 differentially methylated CpGs when comparing the E. coli 3 hpc group and the unchallenged control group on the log 10 scale. The chromosome wide distributions of CpG sites for the comparisons groups were shown were examined by cluster analysis using hierarchical clustering method. The heatmap shows differential methylation of the top 100 CpG sites between different groups , GDNF (cg22964888), PTHR20855_SF21 (cg3759161), MYO1B (cg95955381), GNG7 (cg76083105) and LARP1. Similarly, the top ten differentially methylated loci at the late phase of infection were identified by pairwise comparison of E. coli 24\u2009hpc and control samples. These differentially methylated loci mapped to genes including CYB5R1 (cg24937951), COLA4 (cg77052099), SDF4 (cg63545568), NTN1 (cg54274409), DNAH17 (cg3518148), GSE1 (cg3323937), MMP20 (cg33280374), ABCA4 (cg123320105) and SF3B2 (cg6288374) and late 24 hpc) phases of hpc and hpc phasHomo sapiens UCSC TFBS function, a function not yet supported in Sus scrofa spp. the results revealed the enriched transcription factors identified by comparing E. coli 3 hpc to control samples , CREB , and IRF2 (Interferon Regulatory factor-2). Similarly, E. coli 24\u2009hpc vs unchallenged analysis revealed enrichment of transcription factors , SRXN1 (cg34587223), JAK2 (cg217001619), AQP2 (cg62156997) and ZMYM2 (651169) were selected for bisulfite PCR and pyrosequencing analysis. The results validated the methylation patterns of CpG sites for SRXN1, SENP6, and JAK2 that were hypomethylated in E. coli after 24\u2009h challenged cells compared to the control cells. However, the pyrosequencing validations were not significant for CpG sites in AQP2, and ZMYM2 genes , sulforedoxin (SRXN1), Colony stimulating factor 1 (CSF1), and CXCL14. The zinc finger transcription factor (ZMYM2) gene exhibited no change in expression between when comparing control and E. coli challenged PMEC was also examined. There was a significant reduction in the expression of DNMT3a (de novo methyl transferase) in 3 hpc cells, however no major changes were observed for other related genes (DNMT1 and DNMT3b).The differentially methylated CpG loci within promoter or regulatory regions were deemed quite likely to influence the expression of their downstream genes. We selected genes with the significant hypomethylated CpG sites in the promoter (\u2212\u20092\u2009kb to +\u20091\u2009kb from TSS) as identified in our study for quantitative expression analysis. Significantly higher expression was noticed for stromal derived factor 4 associated mastitis [E. coli-derived pathogenic factor for inflammatory response in mastitis, induces changes in innate immunity genes in dairy cows, and a role for DNA methylation has been demonstrated for this process [mastitis \u201315. The process . Exposur process . Also, a process .E. coli challenge of PMEC resulted in up-regulation of immune response genes, such as chemokines, cytokines, and cell adhesion molecules.Pigs are an important disease model of coliform mastitis, and the present study is the first to investigate the underlying epigenetic mechanisms mediating host-pathogen interaction in cultured porcine mammary cells. To achieve this, we have taken advantage of the clear differences in gene expression patterns occurring during the immediate (3\u2009h) and late immune response (24\u2009h) observed in our earlier studies , 12 wherE. coli challenge. The genome wide methylation patterns indicated comparatively lower CpG methylation in upstream promoter regions compared to gene body regions, and a bimodal distribution of CpG methylation levels was observed. These findings are consistent with prior studies on genome wide CpG methylation in pig [E. coli challenge, respectively at P\u2009<\u00a00.01. Annotation of these CpG probes identified differentially methylated genes, and pathway analysis revealed that the highest enriched functional associations of these genes were inflammatory diseases and organismal injury. In terms of molecular and cellular functions, the top differentially methylated probes associated with cell death & survival, post-translation modifications, cell morphology, cell growth & proliferations. Further differentially methylated CpG\u2019s were clustered into different groups of related genes that indicated immune response as one of the major biological process affected including other cellular processes. Promoter regions are known to have binding sites for different transcription factors that drive gene expression. The genes with differentially methylated CpG\u2019s in their promoters were used to identify relevant transcription factors, and several such identified factors have known prominent roles in inflammation, including PAX5, AP4, CREB, IRF2 and XBP1. PAX5 is a transcription factor that regulates various B cell functions including activation of NF-kB [IRF2 (Interferon regulatory factor 2) regulates IFN-\u03b2 expression and was found to inhibit LPS induced proinflammatory responses [CREB (cAMP-responsive element binding protein) transcriptionally activates pro-inflammatory genes, such as IL-2, IL-6, and TNF-\u03b1 by binding to the cAMP response element (CRE) in their promoter sequences [XBP1 (X-box binding protein 1) is a positive regulator of TLR gene induction and plays a major role in interlinking LPS-associated TLR activation to Endoplasmic Reticulum (ER) stress [In the current study, we examined changes in DNA methylation induced by n in pig , 20. Ourof NF-kB . IRF2 (Iesponses . CREB (cequences . XBP1 (X) stress .SDF4, SRXN1, CSF1, CXCL14 and ZMYM2, all genes which we found to have a significant challenge-dependent reduction in CpG methylation in their upstream regulatory regions around TSS. SDF4, SRXN1, and CXCL14 were selected from hypomethylation CpGs at 24\u2009h after E. coli-challenged, while CSF1 were selected from hypomethylation CpGs at 3\u2009h after E. coli-challenged. The quantitative transcript estimation showed that only the expression of SDF4 and CSF1 in E.coli-challenged cells at 24\u2009h and 3\u2009h was significantly higher than in control cells. However expression pattern and methylation level of SRXN1, and CXCL14 didn\u2019t show a negative relationship. Many studies reported different methylation sites associated with expression, regardless of the directional change in expression and methylation level [ZMYM2 was also higher in the challenged cells, although this difference was not significant. These results confirmed that the CpG methylation in the promoter regions of these genes correlates with changes in gene expression.In general, it is known that hypomethylation of CpG motifs in the promoter regions of genes enhances their transcription. As shown in earlier studies, infection induced hypomethylation of CpGs leads to higher expression of immune genes , 25. In on level . This maon level . The expSDF4 and significantly higher expression of SDF4 in E.coli-challenged PMEC. SDF belong to the CXC subfamily of proteins, are expressed in different tissues, and function as chemokines to attract inflammatory cells [SDF1 (CXCL12) can be seen in inflammatory conditions, such as sub-acromial bursitis and acute liver injury [SDF4 expression was found to be significantly reduced in mammary tumours compared to normal tissue and low levels of SDF4 are linked to a poor prognosis [SDF expression and immune response in mammary tissues.The present study revealed promoter hypomethylation of ry cells . Increasr injury , 30. SDFrognosis . Our resSRXN1 gene had reduced CpG methylation and significantly increased expression in E. coli-challenged cells. Sulfiredoxin (SRXN1) is an antioxidant enzyme that prevents ROS injury to cells, can reduces oxidized cysteine residues of peroxiredoxin proteins (Prx I- IV), and facilitate removal of free radicals [E. coli induces reactive oxygen species (ROS) formation leading to upregulation of antioxidant enzymes, including SRXN1, and SRXN1 production in macrophages was shown to protect mice from LPS induced endotoxic shock [SRXN1 gene has putative NF-kB and AP1 transcription factor binding sites required for up-regulation of SRXN1 expression by LPS treatment [SRXN1 expression.In this study, the upstream regulatory region of the radicals . LPS relic shock . Earlierreatment . HypometCSF1 in the immediate early phase of E.coli challenge but not at the later phase. Colony stimulating factor-1 (CSF-1) has been shown to differentiate peripheral blood monocytes into tissue macrophages [CSF-1 expression is induced by uropathogenic E.coli infection and has a critical role in bacterial clearance during infection [CSF1 corresponds to hypomethylation in the upstream regulatory regions of the CSF1 gene. Curiously, expression of the CSF1 receptor (CSF1R) is also known to be regulated by DNA methylation of its promoter region [CXCL14 is an additional gene found to be regulated in this study. CXCL14 is a chemokine that plays inflammatory modulator and host defence roles in epithelial tissues [Streptococcus pneumoniae [CXCL14 in prostate cancer cells. Treatment of these cells with the demethylating agent 5-aza-2-deoxycytidine affects a hypermethylated CpG island in the CXCL14 gene promoter resulting in the recovery of CXCL14 expression and restoration of chemotaxis [CXCL14 can also be inferred from our study where an E.coli challenge induced hypomethylation in the promoter region of the CXCL14 gene associated with higher levels of expression. These results indicate the potential involvement of epigenetic mechanisms in regulating host cell response to E. coli infection.In the present study, we found increased expression of rophages , 36. CSFnfection , 38. Ther region . CXCL14 tissues , 41. It eumoniae . There iemotaxis . MethylaSome of the genomic regions showed different methylation between infection groups, including CpGs at SSC5 39.78\u201340.07\u2009Mb (OSBPL8) and at SSC9 63.38\u201363.39\u2009Mb (MROH9). These regions (9:63272406\u201363401079\u2009bp and 5: 39774063\u201339828561\u2009bp) contained large CpG islands (CGI) (genome order: Sscrofa11.1). These regions also showed heterogeneity in methylation and changes in the degree of methylation between infection groups. But both of these regions are far away from the promoter site of the transcripts.DNMT3a in the E. coli infected PMEC compared to the control group; however, no changes in the patterns of either DNMT1 or DNMT3B were observed. Hypomethylation in upstream regions of immunity genes may partially be explained by the reduced expression of DNMT3a. It was previously shown that UPEC infections increase DNMT1 expression, the enzyme responsible for maintenance DNA methylation [DNA methyltransferases (DNMT) are the principle enzymes responsible for controlling epigenetic modifications, and DNMT3a and DNMT3b are responsible for de novo DNA methylation. In the present study, the quantitative expression results indicated significantly reduced expression of hylation . The levE. coli challenge in PMEC. CpG methylation changes in the upstream regulatory regions were used to identify enriched transcription factors that regulate immune response pathways. Further, reduced DNA CpG methylation was observed in the immune response genes with corresponding increases in their expression. These results indicate potential epigenetic mechanisms that regulate inflammation during coliform mastitis in pigs.In conclusion, the present study identified first time genome wide differential CpG methylation patterns induced by German Law of Animal Protection guidelines were followed for collecting the tissues. Animal Care Committee at Leibniz Institute of Farm Animal Biology (FBN), Dummerstorf 18196, Germany approved the experiments. The sows were weighed and slaughtered by electronarcosis followed by exsanguination in the experimental abattoir of the FBN. Tissues from mammary complexes cranial of the navel were collected aseptically immediately after slaughter from each individual. After tissue collection all animals underwent routine processes of the slaughterhouse. Veterinary inspection of the animals before slaughtering and of their carcasses and organs after slaughter proofed that they were without any impairment, disease symptoms and pathological signs. Primary cultures of PMEC were obtained as described in our earlier studies . BrieflyE. coli strain isolated from milk of PDS-positive sows described in our previous study [5 PMEC from each 3 sow were seeded and cultured in collagen-coated 6-well plates in complete medium without APS . After 24\u2009h, the medium was changed. Forty-eight hours after seeding, the cells reached ~\u200990% confluency. Then, PMEC were challenged with 107/ml heat-inactivated E. coli for 3\u2009h or for 24\u2009h. The control PMEC cells were not challenged with E. coli. The medium was discarded, and the cells washed three times with phosphate buffered saline to remove bacteria after incubation periods. The experiments included in triplicates for each three animals in three groups . In total 27 genomic DNA and total RNA samples were isolated from treated and control PMEC.The present study uses the same us study , 12. BriE. coli challenged PMEC at two different time points and 24\u2009hpc) and unchallenged control. PMEC were used for libraries construction. RRBS libraries were prepared using 2\u2009\u03bcg of pooled genomic DNA with a 1% spike-in control (unmethylated cl857 Sam Lambda DNA (Promega)). The genomic DNA was digested with Msp I and Taq\u03b1I. Double-enzyme (MspI and Taq\u03b1I) digestion RRBS with increased size-selected fragments will enhance genome-wide CpG coverage. The digested fragments were end repaired, A-tailed and ligated with the C-methylated adaptor sequences TruSeq Nano DNA Sample Preparation kit (Illumina) by following the manufacturer\u2019s protocol . The DNA fragments were later size selected for 40\u2013200\u2009bp with a 2.5% NuSieve 3:1 agarose gel and extracted using the Zyomclean\u2122 Gel DNA Recovery Kit (Zymo Research). The purified DNA was treated with bisulfite using the EZ DNA Methylation-Gold Kit\u2122 (Zymo Research). The preparative scale PCR was performed for 15\u2009cycles and PCR products were purified with the DNA Clean and Concentrator Kit\u2122 (Zymo Research). The qualities of the RRBS libraries were assessed using an Agilent DNA 1000 kit (Agilent Technologies). NGS of the 9 RRBS libraries were performed on an Illumina HiSeq2500 for single-reads of 114\u2009bp at the FBN, Dummerstorf. The bcl2fastq2 conversion software v2.19 was used to convert base call (BCL) files from a sequencing run into FASTQ files that were used for further analysis.Equivalent amount of genomic DNA from three technical replicates per individual animal were pooled. In total 9 pooled samples of DNA, three for each in-silico bisulfite converted porcine genome (11.1) using the Bismark alignment tool . Bisulfite treatment converts unmethylated cytosines to uracils whereas, methylated cytosine is not affected. The sequence reads were mapped to the pre-converted reference genome (Sscrofa 11.1), reads aligned to the multiple regions are removed and best uniquely mapped reads were used for methylation calling.The sequence reads were assessed for quality using FastQC and bases with Phred score greater than 20 were retained for further downstream analysis. RRBS introduces artificial CpG at the 3\u2032 end of the fragments that are removed along with the adaptor sequences to avoid their inclusion in the methylation calling. The default settings for Trim Galore were used for Illumina adaptor trimming as they specifically remove the first two bases from the 3\u2032 end of the sequence such that the additional \u2018C\u2019 closest to the enzyme cut site is removed. The trimmed reads were mapped to the P\u2009<\u00a00.01. A heatmap was used to show the methylation differences between the groups using selected DNA methylation loci. The differentially methylated CpG were annotated to genomic features by using the genomation R/Bioconductor package.Methyl call files from the Bismark aligner with the percent methylation score per base were taken as input files for analysis. The reads which cover all the treatment and control samples with a minimum coverage of 10 were only considered to increase the power of statistical test. The reads from the sex chromosomes, mitochondria, unannotated genome segments and those showing no methylation variation across all samples were filtered out. Differential methylation analysis was done using the MethylKit . LogistiDifferentially methylated CpGs identified by genome-wide analysis were validated using bisulfite PCR and pyrosequencing methods. The same genomic DNA which was used for genome wide methylation analysis was treated with bisulfite using EZ DNA Methylation-Gold Kit\u2122 (Zymo Research). Primers were designed using the Pyrosequencing Assay Design Software with its core analysis features was used. Differentially methylated CpGs present between \u2212\u20092.5\u2009kb and\u2009+\u20091\u2009kb from an annotated transcription start site (TSS) were considered as being in the promoter region of a gene. Such promoter gene IDs were used for transcription factor enrichment, and transcription factor binding site (TFBS) analysis was done using the default parameters of the UCSC_TFBS track of Homo sapiens due to the lack of a Sus scrofa data track and considering that most of the TFBS are conserved.Functional network analysis was done to gain biological insights into top differentially methylated loci between E. coli challenged and control PMEC using the TRI reagent (Sigma-Aldrich) following the manufacturer\u2019s instructions. Isolated RNA was purified by the RNeasy Mini Kit (Qiagen) and DNase I treatment was done to remove the contaminating DNA. First strand cDNA was synthesized using SuperScript III MMLV reverse transcriptase (Invitrogen) with 1\u2009\u03bcg of RNA, 500\u2009ng oligo (dT) and 500\u2009ng random hexamer primers (Promega). Quantitative real-time PCR was performed using the LightCycler\u00ae 480 Real-Time PCR System (Roche Diagnostics). GAPDH and RPL32 were used as internal housekeeping control genes. The sequences of primers for the selected test genes and internal control genes were designed using Primer3 (v.0.4.1) and percentages (B). The genomic distribution of RRBS reads to the porcine genomic CpG island/CpG shore regions in terms of reads fraction (C) and percentages (D). (TIF 855 kb)Additional file 2:Figure S2. Methylation levels of identified CpG sites. The bimodal distribution of CpG methylation was observed in all the samples (A). The methylation levels (%) at different genomic features such as CpG islands, CpG shores (B) and at Promotes, Exons, Introns and Intergenic regions (C) represented. (TIF 1017 kb)Additional file 3:Figure S3.\u2009k-means clustering of differentially methylated genes (CpG in TSS\u2009\u00b1\u20092000) with k\u2009=\u20092 and scaled as Z-score across rows. Top five gene ontology (GO) biological processes derived from each k-means clusters ranked based on the fold enrichment. A) E coli 3 hpc vs control, B) E coli 24\u2009hpc vs control. (TIF 2836 kb)Additional file 4:Figure S4.\u2009k-means clustering of differentially methylated common CpG that were present in both E. coli 3 hpc vs control and E. coli 24\u2009hpc vs control and top five enriched biological biological process. (TIF 1258 kb)Additional file 5:Figure S5. Differentially methylated CpG sites identified between E. coli 3hpc or 24hpc compared to the unchallenged control group from NGS data compare to pyrosequencing including SENP6 (cg90300054), SDF4 (cg63545568), JAK2 (cg217001619), SRXN1 (cg34587223), ZMYM2 (c651169). The y-axis for both box plots represents methylation level. Genes associated with the CpG are given. Box plot represents the range of variation and median value. (TIF 2466 kb)Additional file 6:E. coli 3hpc compared to the unchallenged control group. (XLSX 55 kb)Differentially methylated CpG sites in Additional file 7:E. coli 24hpc compared to the unchallenged control group. (XLSX 75 kb)Differentially methylated CpG sites in Additional file 8:E. coli 3hpc compared to E. coli 24hpc. (XLSX 71 kb)Differentially methylated CpG sites in Additional file 9:List of primers sequences used for quantitative gene expression and pyrosequencing. (DOCX 15 kb)"} +{"text": "Persons with dementia commonly experience mealtime challenging behaviors resulting in negative outcomes. Appropriate caregiver engagement is critical in engaging residents in eating. Current caregiver behavior measures are neither validated nor specific for mealtime care. A feasible and reliable measure to evaluate caregiver engagement during mealtimes is needed. Our team developed the Caregiver Mealtime Engagement Scale (CMES), a 29-item observational measure with good content validity . The CMES includes 24 positive behaviors and 5 negative behaviors . Each item is scored by frequency on a 0 (never) \u2013 3 scale. Total score ranges from 0-87; higher score indicates better engagement. This study aimed to test the CMES\u2019 reliability and validity through a secondary analysis of 87 mealtime video-recorded observations from a hand feeding trial (P30). The sample included 7 residents and 25 staff from 2 nursing homes. The CMES has good internal consistency (Cronbach\u2019s \u03b1 =.775). Inter-rater reliability was good based on ratings of 20 videos by two independent trained coders. Intra-rater reliability was excellent based on ratings of 20 videos by one trained coder at two times (2-3 weeks apart). The CMES demonstrated good convergent validity based on association with the Relational Behavior Scale and Mealtime Relational Care Checklist . Findings support the CMES\u2019 reliability and validity. Future research is needed to test CMES among a larger diverse sample of caregivers in different settings."} +{"text": "Crimean Congo hemorrhagic fever virus (CCHFV) is endemic in South Africa, but whether mild undiagnosed cases occur is unclear. In a seroepidemiologic survey, only 2 of 387 adults considered at risk because of occupational or recreational activities had evidence of previous infection. Seroprevalence in South Africa remains low within the groups investigated. Nairoviridae, genus Orthonairovirus) is a tickborne virus that causes human disease . A questionnaire inquiring about demographic and occupational information and possible risk exposure was completed for each volunteer participant. We collected 374 blood samples from volunteers during April 2016\u2013February 2017 and included 13 stored serum samples, collected mainly from large animal veterinarians in 2012.Specific IgG against CCHFV was detected by using a commercial indirect immunofluorescence assay (IFA) , according to the manufacturer\u2019s instructions. Each IFA slide contains biochips coated with transfected cells expressing either CCHFV glycoprotein (GP), nucleoprotein (NP), or untransfected cells. We screened serum samples at a dilution of 1:100 and retested positive or undetermined samples using serum diluted 2-fold from 1:100 to 1:800. Samples reacting against CCHFV NP only were retested using 2-fold dilutions from 1:10 to 1:80 for evidence of low reactivity against CCHFV GP. We tested all positive reactors for IgM using IFA.Most (299 [77.3%]) participants were from the Free State province ; Figure.Abattoir workers formed the largest high-risk group sampled, accounting for 215 (55.6%) of participants. An additional 30 (7.8%) participants were involved in informal slaughtering. Most participants reported multiple potential routes of exposure, either currently or in the past, resulting in considerable overlap among the different groups. A total of 163 (42.1%) participants reported tick exposure; 27 (7%) participants reported an illness after a tick bite or exposure to animal blood or tissue, and 18 (4.7%) reported a confirmed diagnosis of tick-bite fever.Of the 387 serum samples tested, 2 tested positive for CCHFV IgG. The seropositive samples were collected from men, both 27 years of age, who were abattoir workers at the same abattoir in rural Free State. Both participants had additional potential CCHFV risk exposures, including tick exposure and hunting . NeitherIgG-positive samples for both men tested IgM negative, which excluded acute or recent infections. The IgG titers obtained for participant 1 were 1:100 against the NP and 1:80 against the GP antigen. The IgG titer for participant 2 was 1:400 against the NP antigen only. The variation in antibody titers against NP and GP is not unexpected and has been reported previously, although the reason is unknown. Evidence exists of serologic cross reactivity between CCHFV and Hazara virus; however, previous serologic surveys suggest that Hazara virus is not circulating in South Africa .Our seroprevalence results were similar to those obtained 30 years ago among farm workers (The 2 participants with CCHFV IgG tested negative for CCHFV IgM and recalled no previous illness resembling severe Crimean-Congo hemorrhagic fever, which might hint at possible mild CCHF in South Africa. However, in view of documented widespread CCHFV and antibodies in ticks and animals, respectively, in South Africa ("} +{"text": "OBJECTIVES/SPECIFIC AIMS: Improving human papillomavirus (HPV) vaccination rates ultimately decreases the morbidity and mortality of HPV-associated diseases. A school-based program was piloted in the Rio Grande City Consolidated Independent School District (RGCCISD) to increase HPV vaccination. METHODS/STUDY POPULATION: We assessed baseline HPV vaccination; surveyed 622 parents of eligible children aged \u22659 years; and piloted and developed a school-based HPV education and vaccination program in 1 middle school in 2017 and 4 additional middle schools in 2018. The parent survey included (1) demographic information, (2) an assessment of parental knowledge about the HPV vaccine, and (3) information about their children and HPV vaccine experience. Results of the parent survey and pilot program are in progress. RESULTS/ANTICIPATED RESULTS: As of 9/1/2016, 20.4% of the 7527 RGCCISD eligible students (\u22659 years) had completed the HPV vaccine. Baseline completion rates were higher for RGCCISD students aged 12\u201314 years compared with students aged 9\u201311 and \u226515 years (28.4% vs. 16.5%). Baseline completion rates for RGCCISD adolescents were substantially lower than those reported in NIS-Teen and for Texas . DISCUSSION/SIGNIFICANCE OF IMPACT: Initial results show that engagement with key stakeholders is important and schools are a great venue for delivering and increasing HPV vaccination."} +{"text": "OsPSTOL1) gene through marker-assisted backcross breeding (MABB) in to two intermediate genetic stocks of popular local-varieties namely, ASD 16 and ADT 43 which harbour bacterial blight and blast resistance (R) genes. To delve into the P starvation phenotypic effect, we have generated a set of four backcross inbred lines (BILs) with enhanced P starvation tolerance. The developed BILs showed altered root architecture pattern and greater root surface area with increased P uptake, confirming their adaptability to P deficient soil conditions. Further, a correlation between root traits and low/high P conditions indicates the function of introgressed OsPSTOL1 in BILs. The enhanced root characteristics, therefore, enabled the plants to access and effectively absorb available nutrients from soil. In summary, the unique features of the OsPSTOL1 BILs with bacterial blight and blast resistance can aid varietal development suitable for cultivation in P deficient soils.Phosphorus (P), an essential macronutrient, is a prerequisite for various plant-growth mechanisms including root establishment/development, early/late vegetative stage development and reproductive stage development. Rice (Oryza sativa) is very sensitive to P starvation. Most cultivated genotypes have poor tolerance levels to P deficiency and consequently the grain yield is severely affected by P starvation. Since P deficiency of soils is a major concern of rice production areas, it is necessary to develop new cultivars with enhanced P tolerance. This is also an expectation of farmers and the Agriculture ministry of southern states of India where rice cultivation is intensive. Our objective was to introgress the phosphorus starvation tolerance ( Oryza sativa L.), as one of the major staple crops of the world, feeds billions of people. Increases in the global population demand an equitable rise in grain production. However, a recent report indicates that the current trends in crop yield increases will not meet the future demands. The reasons for low rice productivity are 1) decline in natural resources, including nutrients and water, 2) climate change, 3) poor harvest indices of existing crop varieties, 4) prevalence of pest and diseases, and 5) physical soil deterioration. Thus, sustaining the crop yields under changing climatic conditions has become the focus of major rice breeding programs. One of the most effective strategies is to pyramid multiple stress tolerances (both abiotic and biotic) and resource use efficiencies into elite genetic backgrounds through repeated backcrossing and molecular marker-assisted selection.Rice , a major quantitative trait locus (QTL) on chromosome 12 . Crops were cultivated by adopting normal cultivation practices as mentioned in http://agritech.tnau.ac.in/sri.html.Also, we have evaluated the agronomical performance of improved and parental lines under PYoshida nutrient solution with 1002) and network length (cm) were measured adopting the \u201cGiA Roots\u201d program[Seeds were incubated at 37\u00b0C for 3 days to break the dormancy and sprouted in half strength MS media with 0.3\u201d program.5 conidia/ml & 2% carboxymethyl cellulose (CMS) and plants were covered with polythene sheets during the night time. Scoring of test entries was performed through SES scale and readings on leaf blast severity in test entries were collected at 10-day intervals from 25 to 30 DAS [A uniform blast nursery (UBN) method was adopted for the artificial screening of parental lines and backcrossed lines for their responses to blast disease. Each entry was grown in a row of 70 cm long and 10 cm apart. A local susceptible variety CO39 was raised in between every five lines of BILs and parents. The entire nursery was bordered by a row of the susceptible variety CO39 and excessive nitrogen (100\u2013120 kg N/ha) was given to all the entries. Blast pathogen isolates that were available at the Department of Plant Pathology, TNAU, Coimbatore, India were utilized for inoculation. Nursery beds were smeared with a mixture of blast spore suspension 1 x 10o 30 DAS ,49.8 to 109CFU/ml). Plants were inoculated atthe maximum tillering and booting stages [2F3 lines, recipient parental lines and local susceptible check ADT38 were evaluated along with the resistant control, IRBB60.For screening against BB pathogen, each test entry was raised with a spacing of 20 x 15 cm in two rows of 70 cm length. The nursery was encircled by 3 to 4 border rows of local susceptible variety ADT38. The resistant check variety IRBB60 was raised in between every five lines of BILs and parents. Adequate level of nitrogen 150 kg N/ha) was applied. Peptone sucrose agar was used to sub-culture the pathogen and 48-hour old pure slant culture was used to prepare cell suspension . The data of five plants in each replication were recorded for various agronomic characters viz., days to 50% flowering (DFF), plant height (PH), number of productive tillers per plant (NPT), flag leaf length (FL), flag leaf breadth (FB), panicle length (PL), panicle breadth (PB), shoot biomass per plant (SB), number of filled grains per panicle (NFG), spikelet fertility (SF), 1000 grain weight (GW) and single plant yield (SPY) using the standard evaluation system of rice (IRRI 2002) [In the BCRI 2002) . The graRI 2002) .http://darwin.cirad.fr/).Dendrograms were built based on sequential agglomerative hierarchical nesting (SAHN) based unweight pair group method with arithmetic means (UPGMA) using numerical taxonomy and multivariate analysis system (NTSYS-PC) computer package , to asceF-test and Kolmogorov-Smirnov test, respectively.The gelrite growth data were subjected to single-data correlation analysis.The P-related traits were analysed through Student\u2019s t-test. Equality of variances and normality of variables were justifiedby OsPSTOL1-specific markers , K 29\u20133 showed clear scorable polymorphism between the parents with unambiguous banding pattern when compared to K 29\u20131 and K 29\u20132 was conducted using 523 genome-wide SSR markers. This studyidentified 81 polymorphic markers for CB14002/IR74-Pup1cross and 109 polymorphic markers for CB14004/IR74-Pup1 cross and three bacterial blight resistance genes using sequence tagged site (STS) markers were tested at two P levels (0 and 100%). BC lines showed higher root length when compared to recurrent parents under low P conditions in hydroponic conditions. CB14002IL69 showed higher root weight compared to its recurrent parent at moderate P stress (50% P). In case of total Puptake, CB14002IL69 outperformed the parents under low P condition compared to their respective recipient lines (118 to 119) under low P conditions (50% P). CB14004 IL52 showed higher network depth of 0.997 cm than the recipient parent (0.901 cm). Homozygous lines CB14002 IL16 (54.17 cm) and CB14004 IL52 (49.02 cm) showed increased network length when compared to their respective recipient parents under low phosphorus supply .Two dendrograms (one for CB14002 genetic background and the other for CB14004 background) were generated based on the SSR data gathered in the background selection. Based on the dendrogram generated for CB14002 background, 5 genotypes, including the recurrent parent were grouped into two major clusters. Cluster I comprised CB14002 IL16 and CB14002 IL7 and cluster II comprised recurrent parent CB14002 and other pyramids CB14002 IL38, CB14002 IL69. In cluster II, CB14002 and CB14002 IL69 were adjacent to each othermaking a single line. In case of CB14004 background, there were two major clusters, cluster I and cluster II. Cluster I had CB14004 IL23 and cluster II posses CB14004 IL4, CB14004 IL52, and CB14004, in which CB14004 and CB14004 IL52 formed a single line .2F4 generation was on par with the respective recurrent parent for most of the traits examined recorded 26.65 g/plant. The test entries viz., CB 14002 IL6 & 69 and CB14004 IL4 & 52 had higher grain yields than their respective recurrent parent. The improved lines showed nosignificant variation as compared to recurrent parent in terms of days for flowering, number of productive tillers, shoot biomass, and particularly for distinctness, uniformity and stability (DUS) characters. The genetic distance coefficient on twelve agro-morphologic traits of two pyramids and two parental lines revealed that all the pyramided lines were identical to the recipient parent. CB14002 and CB14004 were clubbed in a single cluster while in another cluster, only a solitary line, the donor parent IR74 Pup1 was accommodated confers a broad spectrum resistance for several races or isolates of M. oryzae [2F3 progenies revealed that all the BC progenies that were screened showed a high level of resistance which proved the efficacy of Pi54 as reported earlier [xa5, xa13,and Xa21) is reported to be effective against bacterial blight isolates in major growing regions of the world [Blast ranks first among the rice diseases affecting rice productivity in Asia because of its wide distribution and high incidence levels during growth seasons once conditions are favorable. It has been already demonstrated that pyramiding multiple blast resistant genes exhibiting resistance to multiple races/pathotypes is a powerful strategy to build broad and durable resistance in new cultivars . The bla. oryzae . In our earlier ,68. Bact earlier . Severalhe world ,40,69. RThis study demonstrated the efficiency of marker-assisted backcross breeding combined with phenotyping in developing rice genotypes with improved disease resistance and enhanced level of tolerance to P starvation. These improved versions have to undergo a rigorous evaluation under field conditions for yield, disease resistance and tolerance for low P to assess its performance over locations in the subsequent years. Upon successful evaluation, these lines are expected toassist the farmers for cultivation in P limited soils.Strategies to develop cultivars with good nutrient use efficiency and durable resistance to pests and diseases are relying on the introgression of multiple genes into susceptible cultivars. Results of this study demonstrated accelerated development of rice genotypes upgraded for disease resistance and nutrient use/uptake efficiency through MABB. The fullest success of this work will provide a roadmap to rice breeders to undertake molecular breeding as a tool for developing resilient rice cultivars possessing disease resistance and P use efficiency. In the future, these lines will be forwarded to yield trials over multiple locations to study the stability in performance for the target traits. Furthermore, these backcrossed lines can be used as donor parents in breeding programmes, andcan also serve as a potential base material for studying host-pathogen interactions combined with P tolerance.S1 Appendix(DOCX)Click here for additional data file.S1 Text(DOCX)Click here for additional data file.S1 FigOsPSTOL1 between the parents.Agarose gel electrophoresis pattern of gene based markers viz., (a) K 29\u20131, (b) K 29\u20132, (c) K 29\u20133 located within (TIF)Click here for additional data file.S2 Fig(TIF)Click here for additional data file.S3 Fig(A)Graphical view of recovery of recurrent parent genome in BC2F2 lines of (i) CB 14002 X IR 74-Pup1 and (ii) CB 14004 X IR 74-Pup1 cross . A, proportion of recurrent parent genome and H, proportion of heterozygous loci. (B) Genetic relatedness analysis of (i) CB14002 and, (ii) CB14004 derived lines through construction of dendrogram using SSR genotyping data.(TIF)Click here for additional data file.S4 FigPup1 (B) CB 14004 X IR 74-Pup1.(A) CB 14002 X IR 74-(TIF)Click here for additional data file.S5 Fig2F2 progenies using markers specific to (a) OsPSTOL1 ; (b) Pi54 (Pi 54 MAS); (c) xa13 (using xa13F and xa13R); (d) Xa21 (using Xa21F and Xa21R and (e) xa5 (using xa5_1F and xa5_1R). A, homozygous recurrent parent allele; H, heterozygous and B, homozygous donor allele respectively.Foreground selection ofBC(TIF)Click here for additional data file.S6 FigPup1 lines, we didnot includ outgroup in the analysis).Numerical values were used to draw the tree in DARwin. Click here for additional data file.S1 Table(PDF)Click here for additional data file.S2 Table(PDF)Click here for additional data file."} +{"text": "Sinorhizobium meliloti species live in a symbiotic relationship with alfalfa plants. We report here the draft genome sequence of S. meliloti strain AK170, recovered from nodules of Medicago orthoceras growing in an area impacted by salinization.Root nodule bacteria of Sinorhizobium meliloti strain AK170 was isolated from nodules of Medicago orthoceras (Kar. & Kir.) Trautv. (syn. Trigonella orthoceras Kar. & Kir.) in the Mugodzhary region in 2002. This region is a part of a modern center of alfalfa intr\u043egressive hybridization in northwest Kazakhstan, which has been suffering from manmade salinization since the 1960s (1e 1960s 13.The culture of AK170 was stored in glycerol at \u221280\u00b0C. A glycerol stock was used to inoculate a tryptone-yeast extract (TY) agar plate ; the resde novo using the SPAdes software version 3.11.1 with default parameters . The NCBI Prokaryotic Genome Annotation Pipeline fiber snap-cap tube using a Covaris S2 instrument. The DNA library was constructed using the dual-index NEBNext multiplex oligos (NEB) and the NEBNext Ultra II DNA library prep kit for Illumina (NEB). This DNA library was sequenced with reagent kit version 3 (600-cycle) on a MiSeq platform (Illumina) at the SB RAS Genomics Core Facility . The entire genome was assembled rameters . Gap clorameters has 100% identity with the corresponding sequences in genomes of Rm1021 (GenBank accession number AL591688) and AK83 (GenBank accession numbers NC_015590 and CP002781). The genome of AK170 does not contain any sequences similar to those which were identified as phage-related genomic islands in Rm1021 (hsd genes encoding a type I restriction-modification system and are known as a part of genomic island Sme19T of Rm1021 (hsdR and hsdM of AK170 showed less than 70% nucleotide similarity with those in Rm1021 and more than 81% with the chromosome sequences of Rhizobium sp. strain TAL182 and the chromosome 2 sequence of Rhizobium sp. strain Y9 (GenBank accession numbers CP021024 and CP018000), respectively.The Sinorhizobium meliloti AK170 is deposited in GenBank under the accession number RDQX00000000. Raw sequencing data are registered in the NCBI SRA database under the accession number SRS3952362. This announcement describes the first version of the genome assembly.The genome sequence of"} +{"text": "TUBA1A p.Arg2His variant. This paper also describes the detailed phenotyping required to aid in the interpretation of novel genomic variants. Variants in GBA1, the gene causing Gaucher Disease (GD), are associated with an increased risk of Parkinson\u2019s Disease (PD) [In this Special Issue we bring together papers demonstrating the need for both detailed genomic and phenotypic studies to aid our scientific and clinical understanding of neurogenetic disorders. Genomic techniques such as genome and exome sequencing are vital tools for diagnosing rare neurogenetic disorders and identifying novel causal genes ,2. In thase (PD) ,5. Gattoase (PD) . Genomicase (PD) . Garcia ase (PD) review t"} +{"text": "Hypoxemic patients undergoing fiber-optic bronchoscopy (FOB) are at risk of worsening of respiratory failure requiring mechanical ventilation due to FOB procedure itself and its complications. As patients with respiratory failure are frequently managed by non-invasive ventilation (NIV); feasibility of FOB through NIV mask has been evaluated in some studies to avoid intubation. We describe here our own case series.Clinical data of 28 FOB done through NIV mask in 27 intensive care unit (ICU) patients over 6 years period at our center was collected retrospectively and analysed.2 71.3\u00b114.2, on NIV and oxygen supplementation) patients. All FOB were done at bedside, 15 of them were given sedation for the procedure. Twenty four patients had bronchoalveolar lavage (BAL); three underwent bronchial biopsies, four brush cytology and seven transbronchial biopsies. In 10 patients lung or lobar collapse was reversed. There was no significant change between pre and post bronchoscopy ABG parameters except for improved post FOB PaO2 (p = 0.0032) and SpO2 (p = 0.0046). One patient (3.57%) developed late pneumothorax and 3 patients (10.7%) had bleeding after biopsy. Prior to bronchoscopy 17 patients were already on NIV. Two patients required mechanical ventilation 6 hours after FOB due to subsequent clinical deterioration but could be weaned off later. One patient died on third day after FOB from acute myocardial infarction.Study comprises 27 hypoxemic :363-367. In non-intubated patients FOB may be done via laryngeal mask airway4 or by intubating them.5 These invasive methods may be avoided by using non-invasive ventilation (NIV) during FOB.18 An alternative technique using high-flow nasal cannula oxygen therapy during FOB is also emerging.19Patients with respiratory failure in intensive care unit (ICU) may require fiber-optic bronchoscopy (FOB). However, FOB itself can lead to hypoxemia, hypotension, tracheobronchial bleeding or cardiac arrhythmias.16 We have been using this technique and our experience over a period of six years is presented here.20Bronchoscopy with NIV support is relatively less used technique with small number of published papers and only one from India.The study population comprised of 27 patients undergoing 28 bedside FOB procedures on NIV during a period of six years in a medical\u2013surgical ICU. Intent of bronchoscopy was therapeutic in 10 patients with atelectasis (four patients with one lung and six with lobar collapse) that were unresponsive to physiotherapy and was diagnostic in the remaining eighteen.2/FiO2 ratio <300, respiratory rate >30 per min and using accessory muscles of respiration but were not candidates for immediate intubation. These 11 patients were put electively on BIPAP, based on clinical judgement, one hour before FOB.Of these 28 patients\u2019 episodes, 17 were already on NIV (16 on BIPAP and one on CPAP). All these patients were observed to desaturate even with a brief discontinuation (<10 minutes) of NIV and were therefore considered unfit for bronchoscopy off NIV. The patient on CPAP was electively switched to BIPAP for FOB. The remaining 11 patients (included four with lung collapse and rest with diffuse lung diseases) had PaO2 above 90% during the bronchoscopy procedure.Noninvasive ventilation was delivered through a tight fitting anaesthesia face mask , kept in place using adjustable straps. This mask was then attached to a catheter mount with inbuilt bronchoscopy port, an Oxygen enrichment connector with inbuilt expiratory port and a single limb NIV circuit that was attached to BiPAP-S/T30 machine . Oxygen 2O and EPAP of 4\u20138 cm H2O. For the remaining 12 (including patient on CPAP) patients\u2019 initial settings were IPAP of 12 cm H2O and EPAP of four cm H2O and this was adjusted based on clinical response. All patients were also put on a backup rate of 12 breaths per min. Arterial blood gas(ABG) analysis was obtained after one hour when stable clinical parameters were achieved for these 12 patients, as also for all other patients, before proceeding to FOB. ABG was repeated one hour after FOB procedure.Sixteen patients were already on BIPAP and they continued on their preexisting setting of IPAP of 12\u201320 cm H15 All patients had bedside ECG, blood pressure and pulse oximetry monitoring during the procedure. A critical care physician provided the sedation and monitored the patient during and after FOB procedure. All bronchoscopies were done by consultant pulmonologists. Bronchoscopy was done through nasal passage in 23 patients and through oral route in five patients (using a bite guard inside the NIV mask). Fiber-optic bronchoscope used (Olympus BF-1T30) had 6.0-mm outer diameter with 2.8 mm instrument channel. Chest X-ray was done in all patients two hours after FOB procedure.All patients received topical anesthesia (10% lignocaine throat spray and 2% lignocaine via bronchoscope). Sedation was used as infusion during 15 bronchoscopies (1% Propofol in 14 patients and Midazolam in one patient).Twenty-eight FOB procedures were done using NIV support. Demographic details are shown in . IndicatCollapsed lung or lobes were successfully opened up in all 10 patients by repeated saline and N-acetyl cysteine instillations and suctioning. Broncheoalveolar lavage (BAL) samples were obtained in 24 patients. Bronchial biopsies were done in three patients, brush cytology in four patients and transbronchial biopsies (TBLB) (without using C-arm) in seven patients.All procedures were well tolerated; no patient needed endotracheal intubation and no hemodynamic instability occurred during FOB. Sedation was well tolerated by patients. Three patients had significant bleeding during FOB that was controlled with bronchoscopic instillation of adrenaline and cold saline. Routine postprocedure chest X-rays after two hours did not reveal any pneumothorax. There was significant improvement in oxygenation noted in post bronchoscopy ABGs in comparison with pre-FOB ABG .All patients on pre-FOB NIV (17 patients) remained on BIPAP afterwards too without requiring intubation. Of the remaining 11 patients, NIV could be discontinued 1\u20132 hours after FOB in eight patients; one of them developed acute respiratory distress six hours after FOB including TBLB. A repeat chest X-ray revealed a delayed pneumothorax requiring emergent intercostal tube drainage and invasive mechanical ventilation. The patient could be successfully extubated after two days. However, a day later he died of myocardial infarction. This patient had history of myocardial infarction five year back.Final bronchoscopic diagnosis for this patient was pulmonary tuberculosis and atypical carcinoid , Case 2.25Fiber-optic bronchoscopy is extensively used in ICUs for diagnosis of pulmonary pathology and mucous plugging. However, bronchoscopy is often associated with temporary changes in gas exchange and lung mechanics.26 In contrast, a 5.7 mm bronchoscope occupies 40% of a 9.0 mm internal diameter (ID) endotracheal tube and 66% of a 7.0-mm ID endotracheal tube27 potentially increasing the work of breathing even more.Fiber-optic bronchoscope occupies about 10% of tracheal cross section in nonintubated patients and this can increase work of breathing.28 PaCO2 increases by about 30% and PaO2 is decreased by 40%.21 Around 200\u2013300 mL of tidal volume is lost during each suctioning period. These changes in blood gases result from decrease in lung volumes and loss of functional gas exchange surface during BAL and reflex bronchospasm due to stimulation of vagal receptors in upper airways. Desaturation during bronchoscopy has also been attributed to upper airway collapse that can be reversed by placement of nasopharengeal airway.29 The delay before normalization of gas exchange varies from about 15 minutes for normal lungs to several hours in those with parenchymal disease.30Significant changes in gas exchange parameters occur during FOB, primarily during suctioning. Hypoxemia during or immediately after taking BAL is most common complication.30 The American Thoracic Society recommends avoiding BAL in patients spontaneously breathing with hypercapnia and/or hypoxemia and who cannot be corrected to at least PaO2 of 75 mm Hg or to SpO2 more than 90% with supplemental oxygen.1Significant cardiopulmonary risk has been reported in up to 13% ventilated patients during bronchoscopy.31 As an extension, it has been used to perform FOB in patients with respiratory failure14 and it has been to shown to help in improving oxygen saturation and arterial oxygen tension and avoid invasive ventilation. Mechanisms for these are likely to be same as for treatment of respiratory failure with NIV. Further as a corollary with obstructive sleep apnea, NIV is likely to counter upper airway collapse during bronchoscopy.33 Moreover, as the presence of the bronchoscope within the trachea reduces its caliber and increases airway resistance and work of breathing, application of NPPV may compensate for this extra-work load, thereby improving the tolerability and safety of the bronchoscopic procedure. Small number of studies, mostly observational, have shown successful use of NIV assisted FOB in COPD,18 immunocompromized patients with pneumonia,5 acute respiratory failure with or without pneumonia in ICU,34 ARDS patients35 and non ICU hypoxemic patients.12 Few randomized control trials of NIV versus Oxygen supplementation8 and NIV versus mechanical ventilation13 have all reported benefit of from using NIV during FOB. Even upper gastrointestinal endoscopy,36 endoscopic retrograde cholangiopancreatography37 and transesophageal echocardiography38 have been successfully done under NIV support. In yet another study COPD patient in exacerbations with mild encephalopathy and inability to clear secretions were randomized to only NIV versus NIV combined with early FOB for bronchial toileting and latter had superior clinical outcomes.18Noninvasive ventilation has been successfully used for respiratory failure \u2013 it decreases work of breathing, improves lung mechanics, recruits closed alveoli, improves blood gases and avoids intubation and mechanical ventilation.34 Protected brushing has been reported by Chiner et al.34 In our study too we have reported obtaining BAL (24 patients), bronchial (three patients) and transbronchial (seven patients) biopsies as well as brushings (4 patients) safely.20 Three patients had significant bleed that was managed successfully while only one patient had (late) pneumothorax.In our study too all 27 patients could successfully undergo 28 FOB procedure with NIV support without need for immediate intubation. This is one of the largest study evaluating feasibility of FOB in patients on NIV support. All published studies have reported NIV supported FOB guided BAL procedures. Only few studies like Maitre et al. (6 patients) and Chiner et al (eight patients) have reported bronchial biopsies while Agarwal R has reported transbronchial biopsies (six patients) during FOB on NIV support.18 The first one needed ventilation after developing a delayed pneumothorax that could be a possible complication of transbronchial biopsy procedure during preceding FOB or simply due to the underlying disease itself. The patient could be successfully extubated two days later after complete resolution of pneumothorax and resultant respiratory deterioration. The second patient needing mechanical ventilation after FOB had bilateral lung contusion after a road traffic accident and had only been subjected to BAL. Worsening of gas exchange is well known after BAL,28 which may or may not be prevented or reversed by application of NIV. This patient too could be extubated later.Two of our patients needed intubation and mechanical ventilation within 24 hours of bronchoscopy as have been reported earlier by other authors.Overall there was improvement in oxygenation although FOB can adversely affect it, perhaps due to inclusion of 10 patients who had resolution of their lung/lobe collapse.12 others too have done FOB with NIV support only for ICU patients.All our patients were admitted in ICU. Bronchoscopy was therapeutic for 10 patients as they succeeded in opening up atelectatic areas. In 18 remaining instances FOB was primarily diagnostic in intent. Except for a few studies39 and during NIV supported FOB.40We used sedation for 15 of 28 FOB procedures and it was well tolerated as has been reported by others in hypoxemic patients in general17 full face mask,18 nasal mask with bite block covered by glove finger through which FOB is introduced,34 helmet mask,9 modified total face mask12 and Boussignac CPAP coupled to a face mask.7 No study till date has compared different types of masks.Different interfaces have been used for FOB on NIV support including oro-nasal mask,17 or dedicated NIV devices34 or even Boussignac CPAP device.7 All approaches seem to work equally well though no study has tried to compare different NIV devices.We used a dedicated NIV device (BiPAP-ST) for our FOB procedures. Other studies have reported using either ICU ventilators to deliver NIVOur study has shown that NIV can facilitate both diagnostic and therapeutic bronchoscopic procedures in selected hypoxemic ICU patients while mostly avoiding intubation. Not only BAL but biopsies and brushing could be done safely. Close monitoring of vital signs is required both before initiation of FOB and for extended periods afterwards as late complications can occur. All bronchoscopies were however performed by experienced pulmonologists while being monitored by intensivists with combined expertise to manage complications of FOB."} +{"text": "Desmodus rotundusendogenous retrovirus (DrERV) QR09 was obtained from a bat tissue sample collected from Desmodus rotundus in the Brazilian rain forest. The complete genome was sequenced using the next-generation sequencing strategy.The strain Desmodus rotundusendogenous retrovirus (DrERV) QR09 was obtained from a bat tissue sample collected from Desmodus rotundus in the Brazilian rain forest. The complete genome was sequenced using the next-generation sequencing strategy. The full-length genome of DrERV QR09 is 8,256 nucleotides in length and showed high similarity with other DrERVs.The strain Endogenous retroviruses (ERVs) are found in a wide variety of hosts and are possibly one of the first circulating viral strains of their respective hosts \u20133. ERVs \u20135\u2013Desmodus rotundusendogenous retrovirus (DrERV) was sequenced from a brain tissue sample of a bat (Desmodus rotundus) collected in the Brazilian Amazon rainforest . The project was approved by the Ethics Committee on the Use of Animals of Evandro Chagas Institute and Biodiversity Information and Authorization System .The strain QR09 of The viral particles were released from the cells using a stainless bead with a TissueLyser II (Qiagen), and the sample was preenriched using 0.45-\u00b5m filters and treatment with benzonase (25 U/liter). DNA and RNA were extracted with a iPrep PureLink virus kit (Thermo Fisher) following the manufacturer\u2019s guidelines. The extracted DNA and RNA were quantified with a Qubit 2.0 fluorometer (Thermo Fisher) using the Qubit RNA HS assay kit, as well as the Qubit double-stranded DNA (dsDNA) HS assay kit (Thermo Fisher). The RNA samples were subjected to reverse transcription using the cDNA synthesis system kit , according to the manufacturer\u2019s guidelines. The cDNA and DNA of brain tissue were combined and sequenced as a single sample. Sequencing libraries were constructed using the Illumina Nextera XT DNA sample preparation kit and sequenced on an Illumina HiSeq 2500 instrument with the high-output V4 2 \u00d7 100-bp sequencing kit .The raw data were filtered for Q30 quality, adapters were removed using Trim_galore pipeline v.0.4.5 , and reaBetaretrovirus, the lineage shows the 4 common genes gag , protease (837 nt), pol , and env . The gag and protease open reading frames did not present any stop codon; however, for the two coding regions pol and env, 2 and 3 stop codons were found, respectively. According to NCBI BLASTn analysis, the DrERV QR09 possesses 99% sequence identity over 100% query coverage of the DrERV isolate 824 (GenBank accession number KP175580) and 99% sequence identity over 96% query coverage of the DrERV isolate 216 (GenBank accession number KP175581); both genomes described were collected in Mexico from D. rotundus.The complete genome of DrERV strain QR09 is 8,256 nucleotides (nt) long, with 65-fold coverage and 48.6% GC content. Compared with the common structure of Desmodus rotundusendogenous retrovirus (DrERV) strain QR09 has been deposited in NCBI GenBank under the accession number MH648003. The sequencing reads (under Sequence Read Archive number SRR8208870) can be accessed through BioProject number PRJNA480298.The complete genome sequence of"} +{"text": "The dyserythropoietic anemia disease is a genetic disorder of erythropoiesis characterized by morphological abnormalities oferythroblasts. This is caused by human gene C15orf41 mutation. The uncharacterized C15orf41 protein is involved in the formation ofa functional complex structure. The uncharacterized C15orf41 protein is thermostable, unstable and acidic. This is associated with TPD domain (135 to 265 residue position) and three PTM sites such as K50 (Acetylation), T114 (Phosphorylation) andK176 (Ubiquitination). C15orf41 is paralogous to isoform-1 (gi|194018542|) and open reading frame isoform-CRA_c (gi|119612744|)of Homo sapiens located at chromosome 15. It interacts with the human ATP (Adenosine Triphosphate) binding domain 4 (ATPBD4)having similarity score 0.725 as per protein-protein interaction (PPI) network analysis. This data provides valuable insights towardsthe functional characterization of human gene C15orf41. The human uncharacterized gene C15orf41 is located atchromosome 15 encodes a protein with two predicted helix-turnhelixdomains. Mutations of this gene are found in the family ofcongenital dyserythropoietic anemia type-I [The structural and functional characteristics of proteins play thesignificant role in drug design and discovery. Investigations ofthese proteins characteristics experimentally in the wet lab aremuch laborious, time consuming and costly. Thecomputational/statistical tools of bioinformatics reduce this costand time significantly to characterize the uncharacterizedproteins. These tools are widely used for homology modeling ofsequence profiles and predicting the three-dimensional 3D)structure of the targeted protein. The homology modeling isutilized when the experimentally obtained structure isunavailable. It can provide a useful 3D model for the protein ofinterest that is related to at least one known protein structure. It isalso used to predict the 3D structure of one or more proteins ofknown structure for a given protein sequence based on theprimarily sequence alignment. The inclusive municipal sequencesare increasing in some databases like SwissProt and NCBIDstructuThe target/query sequence (human uncharacterized proteinC15orf41) was collected in FASTA format using the accessionnumber Q9Y2V0 from the UniProt protein database(http://www.uniprot.org/uniprot/Q9Y2V0) .ReferenHomology modelling are used to identify the structure of thequery protein sequence based on one or more known proteinstructures and on the production of an alignment that mapsresidues in the query sequence to residues in the templatesequence. The quality of the homology model is associated withthe quality of the sequence alignment approach and templatestructure. Homology model produces high-quality structural models when the target and template are closely related.Homology search of the target/query protein sequence with thereference/template sequences are performed using the onlinebioinformatics tool \"BLASTp \" of NCBI database .Various physiochemical properties of the query protein like thenumber of amino acids, molecular weight, hypotheticalisoelectric point (pI), amino acid composition (%), number ofpositively (Arg + Lys) and negatively charged (Asp + Glu)residues, extinction coefficient, instability index, aliphatic indexand Grand Average of Hydropathicity (GRAVY) are investigatedusing the online ExPASy's ProtParam tool(http://expasy.org/tools/protparam.html).A protein consists of one or more domains for functionalactivities in the cellular processes. Some domains show theirfunctions regularly and some are active during the evolutiononly. The post translational modification (PTM) is themodification of amino acid covalent based on the proteinsequence and it is important issues for regulating of biologicaland physiological functions in the cell . The queThe computational/statistical offline bioinformatics tool is usedThe secondary structure of the query protein sequence ispredicted using the online bioinformatics tool SABLE(http://sable.cchmc.org/) . Then itProtein-protein interaction (PPI) networks analysis of a queryprotein with the template/reference proteins is important formore accurate prediction of its function. The PPI network isperformed using online STRING (http://string-db.org/)bioinformatics tool .The detail workflow of this study is shown in Structural and functional characteristics of human proteinC15orf41 (query protein) was analyzed using the several onlineand offline bioinformatics tools. At first, we performed thehomology modeling of query protein with thereference/template protein sequences using the BLASTp toolfrom NCBI database. Out of which, the best homology(template/reference) protein sequences were selected based onthe different criteria such as maximum score (580), querycoverage (100%), identity (99%) and e-value (0.0), and selectedprotein sequences werre used for the further analyses. Thecomputation of amino acid composition of query proteinsequence using ExPASY's ProtParam tool detected very highpercentages of isoleucine (7.5%) and leucine (11.4%) as comparedto other amino acids of this In this study, suggests that the physiochemical properties of thequery protein seem to be thermostable, unstable and acidic. Wefound that the query protein has three PTM sites such as K50(Acetylation), T114 (Phosphorylation) and K176 (Ubiquitination).The functional similarity is the isoform-1 and isoform-CRA_c ofHomo sapiens, Gorilla (C15orf41) and Chimpanzee(C15H15orf41) with the query protein based on the phylogeneticanalysis. The study protein showed the similar functionalbehavior compare with the known proteins functionality ofGorilla and Chimpanzee. The human ATPBD4 is the highinteracting score protein with the query protein. Thiscomputational study would be helpful for theresearchers/scientists/biologists to characterize the otheruncharacteristic proteins.The authors declare that they have no competing interests."} +{"text": "Adults with cystic fibrosis (CF) have been reported to be at five to ten-fold risk (25 to 30 fold risk after solid organ transplant) of colorectal cancer (CRC) than the general population. Limited publications to date have reported on practical aspects of achieving adequate colonic cleanse producing good visualisation. In this study, we compared two bowel preparation regimens, standard bowel preparation and a modified CF bowel preparation.A non-randomised study of adults with CF attending a single centre, requiring colonoscopy investigation were selected. Between 2001 and 2015, 485 adults with CF attended the clinic; 70 adults with CF had an initial colonoscopy procedure. After five exclusions, standard bowel preparation was prescribed for 27 patients, and modified CF bowel preparation for 38 patients. Demographic and clinical data were collected for all consenting patients.p\u2009=\u20090.006). Rates of \u201cfair\u201d GI cleanse visualisation were similar between the two groups (39.4% versus 29.6%) . Positive adenomatous polyp detection rate in patient\u2019s age\u2009>\u200940\u2009years of age was higher (62.5%) than those <\u200940\u2009years of age (24.3%) (p\u2009=\u20090.003). Colonic adenocarcinoma diagnosis was similar in both groups.There was a significant difference between modified CF bowel preparation group and standard bowel preparation group in bowel visualisation outcomes, with the modified CF bowel preparation group having a higher proportion of \u201cexcellent/good\u201d GI visualisation cleanse (50.0% versus 25.9%) and lower rates of \u201cpoor\u201d visualisation cleanse (10.5% versus 44.5%) than standard bowel preparation\u00a0 was evident. These results support adults with CF considered for colonoscopy screening at 40\u2009years of age, or prior to this if symptomatic; which is earlier than CRC screening in the non-CF Australian population.The online version of this article (10.1186/s12876-019-0979-z) contains supplementary material, which is available to authorized users. Cystic fibrosis (CF) affects more than 70,000 people globally, and adults outnumber children living with CF in many countries . Cystic The detection of CRC by symptoms in the general population, can lead to delayed diagnosis and many countries have programs to assist early detection , 8. AustOptimal CRC screening has not been established in patients with CF, although the CF CRC screening consensus recommendations 2018 have recently been published . These rOver several years CF, transplant and gastroenterology services at The Prince Charles Hospital (TPCH), have observed a higher incidence of inadequate bowel cleanse with standard bowel preparation lavage regimens . In thisn\u00a0=\u200927) and modified cystic fibrosis bowel preparation (n\u00a0=\u200938).A non-randomised cohort study of adults with CF, requiring colonoscopy investigation from TPCH; were selected. Between 2001 and 2015, 485 adults with CF attended our public government funded health system clinic. The state lung transplant service is also based at TPCH. The population included 70 adults with CF who had an initial colonoscopy procedure between 2001 and 2015. Five patients were excluded due to inability to determine the quality of the bowel preparation from the report or accompanying pictures. Standard bowel preparation was performed on 27 patients from 2001 to 2009, (data was collected retrospectively in this group after informed written consent obtained). A modified CF bowel preparation was prescribed after 2009 and informed written consent and all prospective data collected on 38 patients. Additional file Patients provided informed written consent for colonoscopy procedure for investigation of significant GI symptoms including rectal bleeding, constipation, and a family history of colonic cancer or as a part of lung transplant assessment procedures and written consent for data collection as part of this study. Patients requiring repeat surveillance colonoscopy procedures had data recorded on type of preparation and clearance reports. In four cases the initial colonoscopy was extended by one litre of GI lavage (due to inadequate bowel preparation) within 24\u2009h of the initial procedure to permit a full colonoscopic investigation at the clinical discretion of the Gastroenterologist; such examinations within 24\u2009h were counted as one initial colonoscopy procedure.Colonoscopies performed prior to November 2009 included patients with CF who received standard bowel preparation (control group) with data collated retrospectively from medical records and gastroenterology database reports, written consent for data collection was obtained. The modified CF bowel preparation [Additional file The efficacy of the bowel preparation lavage was recorded using the Queensland Bowel Cancer Screening Program Bowel Preparation Descriptor scale modifiedp-value of <\u20090.05 was considered significant. Data analysis was performed using PASW, version 18 .The primary outcome measure was efficacy of bowel preparation lavage documented on the colonoscopy report. Secondary outcome measures were the number of adenomatous polyps and colonic adenocarcinomas detected; duration of pre- and post-colonoscopy hospital length of stay. Student t-tests were used to compare continuous variables and Chi squared test (or Fishers exact test where appropriate) for categorical comparisons. A Baseline characteristics at first colonoscopy procedure were similar between the two groups . Rates of \u201cfair\u201d GI cleanse were similar between the two groups (39.4% versus 29.6%) and lower rates of \u201cpoor\u201d cleanse in the modified CF bowel preparation group than standard bowel preparation (10.5% versus 44.5%)\u00a0(p\u2009=\u20090.01) than those <\u200940\u2009years of age (24.3%) (.5% than Repeat surveillance colonoscopes (20 procedures) in the CF Preparation cohort had similar rates of excellent/good bowel cleansing was 2\u2009days and the median days hospitalised after the colonoscopy was 1\u2009day . By comparison, the median days hospitalised before the colonoscopy (modified CF bowel preparation) was 3\u2009days and the median days hospitalised after the colonoscopy was 1\u2009day .There were no serious adverse events, e.g. deaths, colon perforations, ventilation or intensive care unit (ICU) admissions related to the colonoscopy procedure.Increasing longevity of adults with CF reflects enhanced medical patient management and has led to greater numbers of adults with CF reaching middle age; with further growth expected over the coming decade . The preThe CFF emphasisIn the general population, up to 25% of all colonoscopies are reported to have an inadequate bowel preparation , 13 the To date, limited publications have reported on practical aspects of colonic lavage in adults with CF. In our study, we report poor colonic lavage in 44% of patients with CF undergoing a standard colonoscopy preparation, yet this was reduced to 10% with the use of the longer modified CF bowel preparation. No difference was seen comparing efficacy of colonic lavage from initial and repeated surveillance procedures using the modified CF bowel preparation protocol. Research at another CF center has reported standard colonoscopy preparation protocol to be suboptimal and therefore lead to development of a CF-specific colonoscopy preparation in Minnesota USA ; althougInadequate preparation in the general population has been associated with reduced adenoma detection rates . Our stup\u2009=\u20090.003), supporting this earlier recommendation to initiate colonoscopy screening at age 40 for people with CF [Our centre reported positive adenomatous polyp detection on initial scope rate of 50% (mean age 37.6\u2009years) similar in incidence (49%) to the Minnesota CF cohort Additional file 2:Table S2. Comparison of age and rate of adenomatous polyp detection on first colonoscopy. (DOC 28 kb)Additional file 3:Table S3. Comparison of initial and repeat scopes for efficacy of GI cleanse. (DOC 29 kb)Additional file 4:The Adult Cystic Fibrosis Centre. The Prince Charles Hospital. (TPCH) Brisbane. The Adult Cystic Fibrosis Centre. The Prince Charles Hospital. (TPCH) Brisbane. Modified CF Bowel Preparation. (DOC 34 kb)Additional file 5:Dept of Gastroenterology, The Prince Charles Hospital (TPCH) Brisbane. Preparation for Colonoscopy. STANDARD Bowel Preparation. (DOC 96 kb)Additional file 6:Comparison of Bowel Preparations: 1-The Prince Charles Hospital (TPCH) Brisbane Australia Standard bowel preparation, 2- The Prince Charles Hospital (TPCH) Modified Cystic Fibrosis (CF) bowel preparation and 3 \u2013 University of Minnesota Colonoscopy Prep Instructions for patients with Cystic Fibrosis. (DOC 45 kb)"} +{"text": "This article has been corrected: In the Abstract section, the 2nd sentence of the 3rd paragraph has been corrected to include the following: \u201cPMCA expression was lower in untreated malignant cells than normal cells. CaEP caused decreased expression of NCX1 in malignant cells and RyR1 in both cell lines whereas normal cells exhibited increased expression of NCX1 after CaEP.\u201dCalcium electroporation (CaEP) describes the use of electric pulses (electroporation) to transiently permeabilize cells to allow supraphysiological doses of calcium to enter the cytosol. Calcium electroporation has successfully been investigated for treatment of cutaneous metastases in a clinical study. This preclinical study explores the possible use of calcium electroporation for treatment of sarcoma.in vivo in immuno-deficient mice.A normal murine muscle cell line (C2C12), and a human rhabdomyosarcoma cell line (RD) were used in the undifferentiated and differentiated state. Electroporation was performed using 8 pulses of 100 \u00b5s at 600\u20131000 V/cm; with calcium . Viability was examined by MTS assay, intracellular calcium levels were measured, and expression of plasma membrane calcium ATPase (PMCA) was investigated using western blotting. Calcium/sodium exchanger (NCX1), ryanodine receptor (RyR1) expression and cytoskeleton structure (zyxin/actin) were assessed by immunofluorescence. CaEP efficiency on RD tumors was tested CaEP was significantly more efficient in RD than in normal cells. PMCA expression was lower in untreated malignant cells than normal cells. CaEP caused decreased expression of NCX1 in malignant cells and RyR1 in both cell lines whereas normal cells exhibited increased expression of NCX1 after CaEP. Calcium electroporation also affected cytoskeleton structure in malignant cells.This study showed that calcium electroporation is tolerated significantly better in normal musclreatment this could potentially be used in connection with sare cells than sarcoma cells and as an inexpensive and simple cancer tcoma surgery for local treatment.11604-11618.https://doi.org/10.18632/oncotarget.24352Original article: Oncotarget. 2018; 9:11604\u201311618."} +{"text": "Scragg reports increasing evidence for differences in the thresholds for serum 25-hydroxyvitamin D [25(OH)D] concentration [vitamin D repletion] that need to be reached in populations observationally, or to be achieved by vitamin D supplementation in deficiency, before health benefits become apparent in different conditions . Such va3 intakes and serum 25(OH)D concentrations, which would allow rationalisation of public health decisions on recommended daily intakes, according to demonstrable biological effects of different levels of vitamin D status in humans. Reductions in abnormal insulin resistance have been reported after six months of supplementation with vitamin D at 4000 IU/day in deficient South-East Asian women living in New Zealand, but only in those women who achieved 25(OH)D values >80nmol/L and betw"} +{"text": "Anaplastic meningioma is a rare and aggressive brain tumor characterised by intractable recurrences and dismal outcomes. Here, we present an integrated analysis of the whole genome, transcriptome and methylation profiles of primary and recurrent anaplastic meningioma. A key finding was the delineation of distinct molecular subgroups that were associated with diametrically opposed survival outcomes. Relative to lower grade meningiomas, anaplastic tumors harbored frequent driver mutations in SWI/SNF complex genes, which were confined to the poor prognosis subgroup. Aggressive disease was further characterised by transcriptional evidence of increased PRC2 activity, stemness and epithelial-to-mesenchymal transition. Our analyses discern biologically distinct variants of anaplastic meningioma with prognostic and therapeutic significance. Nearly half of anaplastic meningiomas represent progression of a previously resected lower grade tumor, whereas the remainder arise de novo4. Recurrence rates are 5\u201320% and 20\u201340%, respectively, for grade I and II tumors5. By contrast, the majority of anaplastic meningioma patients suffer from inexorable recurrences with progressively diminishing benefit from repeated surgery and radiotherapy and 5-year overall survival of 30\u201360%6.Meningiomas arise from arachnoidal cells of the meninges and are classified as grade I (80% of cases), grade II (10\u201320%) or grade III (1\u20133%). Grade III meningiomas comprise papillary, rhabdoid and anaplastic histological subtypes, with anaplastic tumors accounting for the vast majority of grade III diagnoses7. In keeping with previous smaller studies, mutually exclusive mutations in NF2 and TRAF7 were the most frequent driver events, followed by mutations affecting key mediators of PI3K and Hedgehog signalling8. Recurrent hotspot mutations were also identified in the catalytic unit of RNA polymerase II (POLR2A) in 6% of grade I tumors7. More recently, a study comparing benign versus de novo atypical (grade II) meningiomas found the latter to be significantly associated with NF2 and SMARCB1 mutations9. Atypical meningiomas were further defined by DNA and chromatin methylation patterns consistent with upregulated PRC2 activity, aberrant Homeobox domain methylation and transcriptional dysregulation of pathways involved in proliferation and differentiation9.A recent study of 775 grade I and grade II meningiomas identified five molecular subgroups defined by driver mutation profile10. Here, we present an analysis of the genomic, transcriptional and DNA methylation patterns defining anaplastic meningioma. Our results reveal molecular hallmarks of aggressive disease and suggest novel approaches to risk stratification and targeted therapy.Despite the high mortality rate of anaplastic meningiomas, efforts to identify adjuvant treatment strategies have been hampered by a limited understanding of the distinctive molecular features of this aggressive subtype. A recent analysis of meningioma methylation profiles identified distinct subgroups within Grade III tumors predictive of survival outcomes, though the biology underpinning these differences and any therapeutic implications remain unknownWe performed whole genome sequencing (WGS) on a discovery set of 19 anaplastic meningiomas resected at first presentation (\u2018primary\u2019). A subsequent validation cohort comprised 31 primary tumors characterised by targeted sequencing of 366 cancer genes. We integrated genomic findings with RNA sequencing and methylation array profiling in a subset of samples in the 18 primary tumor genomes , the somatic point mutation burden of primary anaplastic meningioma was low with a median of 28 somatic coding mutations per tumor sequencing of 31 anaplastic meningioma samples from a total of 28 patients (26 primary tumors and 5 recurrences). Gene expression variability within the cohort did not correlate with clinical parameters including prior radiotherapy, anatomical location or clinical presentation (.0) Fig.\u00a0. The sub.0) Fig.\u00a0.Figure 210. Unsupervised hierarchical clustering using methylation data available for a subset of the cohort (n\u2009=\u200919) demonstrated segregation into two main groups largely overlapping the subgroups delineated on the basis of gene expression profile, though correlation with survival outcomes was less marked and upregulation of CXCL14, both prognostic biomarkers in diverse other cancers and function in a paracrine manner31. It is hence possible that some of the gene expression patterns we observed may reflect differences in the tumor stromal compartment, itself an increasingly recognised therapeutic target33.Nineteen hundred genes underpinned the differentiation of anaplastic meningioma into subgroups C1 and C2, which could be reduced to only 6 transcripts selected on the basis of PCA coefficient and differential expression analysis .The C1 tumors were further characterised by upregulation of transcriptional programs associated with increased proliferation, PRC2 activity and stem cell phenotype . In keeping with this finding, formal pathway analysis identified significant dysregulation of stemness, proliferation, EMT and PRC2 activity (Supplementary Tables\u00a042. Yes-associated protein 1 (Yap1), a cornerstone of oncogenic Hippo signalling, is frequently overexpressed in cancer and synergises with Wnt signalling to induce EMT44. YAP1 was upregulated in anaplastic tumors along with MYL9, a key downstream effector essential for Yap1-mediated stromal reprogramming subgroup, which was further characterised by transcriptional signatures of PRC2 target activation, stemness, proliferation and mesenchymal differentiation. These findings were in part underpinned by differential expression of Hox genes. Acquisition of invasive capacity and stem cell traits are frequently co-ordinately dysregulated in cancer, often through subversion of Hox gene programs integral to normal tissue morphogenesis52. Hox genes have a central role in orchestrating vertebrate development and act as highly context-dependent oncogenes and tumor suppressors in cancer53. Several of the most starkly upregulated Hox genes in the C1 tumors consistently function as oncogenes across a range of solid and haematological malignancies, including HOTAIR, HOXB7, HOXA4, HOXA-AS2, HOXC11, and NKX2-262. Like many other long non-coding RNAs (lncRNA), HOTAIR and HOXA-AS2 modulate gene expression primarily by interacting directly with chromatin remodelling complexes, exerting oncogenic activity by recruiting PRC2 to target genes65. HOXA-AS2 has been shown to mediate transcriptional repression of the tumor suppressor gene CDKN2A (p16INK4A), deletion of which is associated with poor meningioma survival67. Given the antagonistic relationship between the SWI/SNF and PRC2 chromatin regulators, deleterious SWI/SNF mutations and overexpression of lncRNAs known to mediate PRC2 activity emerge as potentially convergent mechanisms underpinning the differences between C1 and C2 tumors68. Further endorsing a link between transcriptional subgroups and chromatin dysregulation, 15 of the differentially expressed transcripts delineating C1 and C2 subgroups (absolute log2 fold change >2 and FDR\u2009<\u20090.01) are among the 50 genes most often associated with frequently bivalent chromatin segments (FBS) in cancer, including 11 transcripts from the HOXB cluster on chromosome 1769. This overlap was highly statistically significant (hypergeometric distribution P\u2009=\u20091.98\u2009\u00d7\u200910\u221211). Bivalent, or epigenetically \u2018poised\u2019, chromatin is characterised by finely balanced activating (H3K4me1/H3K4me3) and repressive (H3K27me3) histone marks and pre-loaded DNA polymerase II poised to transcribe in response to modest epigenetic changes70. Bivalent chromatin most often marks genes involved in developmental reprogramming, in particular Hox cluster genes and homeotic non-coding transcripts, and is a frequent target of aberrant chromatin modification in cancer71.Although anaplastic tumors resist comprehensive classification based on driver mutation patterns, transcriptional profiling revealed two biologically distinct subgroups with dramatically divergent survival outcomes. This finding is emblematic of the limitations of histopathological grading as a risk stratification system for meningiomaSMARCB1 mutations and large deletions encompassing chromosomes 1q, 6q and 14q. Notably, these genomic regions encompass ARID1A and several other SWI/SNF subunit genes. Both SMARCB1 mutations and the aforementioned copy number changes were associated with epigenetic evidence of increased PRC2 activity, differential Homeobox domain methylation, and upregulation of proliferation and stemness programs in atypical grade II meningiomas9.In the context of recent studies of lower grade meningiomas, our findings raise the possibility that the balance between PRC2 and SWI/SNF activity may have broader relevance to meningioma pathogenesis. Compared to grade I tumors, atypical meningiomas are more likely to harbor 74. Several EZH2 inhibitors are in development with promising initial clinical results75. Other modulators of PRC2 activity, including HOTAIR, may also be relevant therapeutic targets77. Furthermore, growing recognition of the relationship between EMT and resistance to conventional and targeted anti-cancer agents has profound implications for rational integration of treatment approaches33. Notably, EGFR inhibition has yielded disappointing response rates in meningioma despite high EGFR expression78. A mesenchymal phenotype is strongly associated with resistance to EGFR inhibitors in lung and colorectal cancer81. Combining agents that abrogate EMT with other therapies is a promising strategy for addressing cell-autonomous and extrinsic determinants of disease progression and may warrant further investigation in meningioma33.The extent to which SWI/SNF depletion plays a role in meningioma development may be therapeutically relevant. Diverse SWI/SNF mutated cancers exhibit dependence on both catalytic and non-catalytic functions of EZH2, a core subunit of PRC2This study has revealed biologically and prognostically significant anaplastic meningioma subgroups and identified potentially actionable alternations in SWI/SNF genes, PRC2 activity and EMT regulatory networks. However, a substantially larger series of tumors, ideally nested in a prospective multicentre observational study, will be required to expand upon our main findings and explore mechanistic and therapeutic ramifications of meningioma diversity.DNA was extracted from 70 anaplastic meningiomas; 51 samples at first resection (\u2018primary\u2019) and 19 from subsequent recurrences. Matched normal DNA was derived from peripheral blood lymphocytes. Written informed consent was obtained for sample collection and DNA sequencing from all patients in accordance with the Declaration of Helsinki and protocols approved by the NREC/Health Research Authority (REC reference 7/YH/0101) and Ethics Committee at University Hospital Carl Gustav Carus, Technische Universit\u00e4t Dresden, Germany (EK 323122008). Samples underwent independent specialist pathology review (V.P.C and K.A). DNA extracted from fresh-frozen material was submitted for whole genome sequencing whereas that derived from formalin-fixed paraffin-embedded (FFPE) material underwent deep targeted sequencing of 366 cancer genes.NAB2-STAT6. This fusion is pathognomonic of meningeal hemangiopericytoma, now classified as a separate entity, solitary fibrous tumors84. We therefore excluded three samples from this tumor from further study. A second sample (PD23354a), diagnosed as an anaplastic meningioma with papillary features, was found to have a strong APOBEC mutational signature as well as an EML4-ALK gene fusion .One tumor sample PD23348 (and two subsequent recurrences) separated from the main study samples in a principal components analysis of transcriptomic data , or 75 base (transcriptomic) paired-end sequencing were performed on Illumina X10 genome analyzers in accordance with the Illumina Genome Analyzer operating manual. The average sequence coverage was 65.8X for tumor samples and 33.8X for matched normal samples to enrich for all coding exons of 366 cancer genes 88. Transcripts Per kilobase per Million reads (TPM) were generated using an in-house python script (https://github.com/TravisCG/SI_scripts/blob/master/tpm.py)88. We downloaded archived RNA sequencing FASTQ files for 19 grade I meningioma samples representing the major mutational groups (ArrayExpress: GSE85133)7. Reads were then processed using STAR and HTseq as described above. Cancer cell line (n\u2009=\u2009252) and triple-negative breast cancer (n\u2009=\u2009100) RNA sequencing data was generated in-house by the aforementioned sequencing and bioinformatic pipeline.For transcriptome sequencing, 350\u2009bp poly-A selected RNA libraries were prepared on the Agilent Bravo platform using the Stranded mRNA library prep kit from KAPA Biosystems. Processing steps were unchanged from those specified in the KAPA manual except for use of an in-house indexing set. Reads were mapped to the GRCh37 reference genome using STAR (v2.5.0c)https://github.com/cancerit/cgpRna)90. Fusions identified by one or two algorithms or also detected in the matched normal sample were flagged as likely artefacts. Fusions were further annotated according to whether they involved a kinase or known oncogene and whether they occurred near known fragile sites or rearrangement break points91 , STAR-Fusion (v0.1.1) and deFuse (v0.7.0) 95.The DESeq2 R package was used for all differential gene expression analyses97. RUVg estimates the factor attributed to spurious variation using control genes that are assumed to have constant expression across samples100. We selected control genes on the basis of previous studies of suitable control genes for transcript-based assays in meningioma101. PEER is based on factor analysis methods that infer broad variance components in the measurements. PEER can find hidden factors that are orthogonal to the known covariates. We applied this feature of PEER to remove additional hidden effect biases. The final fitted linear regression model consists of the factor identified by RUVg method that represents the unwanted laboratory batch effect and 13 additional hidden factors found by PEER that are orthogonal to the estimated laboratory batch effect. Using this approach we were able to reduce the number of DEGs from more than 18000 to 8930, of which <4,000 are predicted to be protein-coding.Preliminary comparison of anaplastic and externally-generated grade I meningioma data revealed evidence of laboratory batch effects, which we mitigated with two batch-correction methods: RUVg and PEER106. We further corroborated these findings with a more general Gene Ontology (GO) pathway analysis107.To identify biological pathways differentially expressed between meningioma grades and anaplastic meningioma subgroups we applied a functional class scoring algorithm using a collection of 461 published gene sets mapped to 10 canonical cancer hallmarks (Supplementary Table\u00a095. PCA was performed using the top 500 most variably expressed transcripts and the R stats::prcomp function108. Given that primary component 1 (PC1) was the vector most clearly distinguishing the closely clustered C2 subgroup from the more diffusely clustered C1 between i) the C1 and C2 anaplastic meningioma subgroups and ii) the C1 anaplastic meningiomas and the 19 grade I tumors function implemented by the DESeq2 package C1 Fig.\u00a0, we extr109. CaVEMan (Cancer Variants Through Expectation Maximization: http://cancerit.github.io/CaVEMan/) was used for calling somatic substitutions. Small insertions and deletions (indels) in tumor and normal reads were called using a modified Pindel version 2.0. (http://cancerit.github.io/cgpPindel/) on the NCBI37 genome build111. Annotation was according to ENSEMBL version 58. Structural variants were called using a bespoke algorithm, BRASS (https://github.com/cancerit/BRASS) as previously described112.Genomic reads were aligned to the reference human genome (GRCh37) using the Burrows-Wheeler Aligner, BWA (v0.5.9)113.The ascatNGS algorithm was used to estimate tumor purity and ploidy and to construct copy number profiles from whole genome data22.To identify recurrently mutated driver genes, we applied an established dN/dS method that considers the mutation spectrum, the sequence of each gene, the impact of coding substitutions and the variation of the mutation rate across genes114. Variants were screened against lists of somatic mutations identified by a recent study of 11,119 human tumors encompassing 41 cancer types and also against a database of validated somatic drivers identified in cancer sequencing studies at the Wellcome Trust Sanger Institute copy number variants meeting these criteria were considered potential drivers. Additional truncating events in established tumor suppressors were also flagged as potential drivers. Only rearrangements with breakpoints able to be reassembled at base pair resolution are included in this dataset.12. TraFiC uses paired-end sequencing data for the detection of somatic insertions of transposable elements (TEs) and exogenous viruses. The identification of somatic TEs is performed in three steps: (i) selection of candidate reads, (ii) transposable element masking, (iii) clustering and prediction of TE integration sites and (iv) filtering of germline events12.For the identification of putative solo-L1 and L1-transduction integration sites, we used the TraFiC (Transposome Finder in Cancer) algorithmWe performed quantitative methylation analysis of 850,000 CpG sites in 25 anaplastic meningiomas. Bisulfite-converted DNA (bs-DNA) was hybridized on the Ilumina Infinium HumanMethylationEPIC BeadChip array following the manufacturer\u2019s instructions. All patient DNA samples were assessed for integrity, quantity and purity by electrophoresis in a 1.3% agarose gel, picogreen quantification and Nanodrop measurements. Bisulfite conversion of 500\u2009ng of genomic DNA was done using the EZ DNA Methylation Kit (Zymo Research), following the manufacturer\u2019s instructions. Resulting raw intensity data (IDATs) were normalized using the Illumina normalization method developed under the minfi R\u00a0package (v1.19.10). Normalized intensities were then used to calculate DNA methylation levels . We then\u00a0excluded from the analysis the positions with background signal levels in methylated and unmethylated channels (p\u2009>\u20090.01). Finally we removed probes with one or more single nucleotide polymorphisms (SNPs) with a minor allele frequency (MAF) >1% in the first 10\u2009bp of the interrogated CpG, as well as the probes related to X and Y chromosomes. From the filtered positions, we selected only CpG sites present both in promoter regions and CpG islands .116. The gene sets used were: H: hallmark gene sets, BP: GO biological process, CC: GO cellular component, MF: GO molecular function and C3: motif gene sets (http://software.broadinstitute.org/gsea/msigdb/collections.jsp). The gene clusters resulting from the hypergeometric test with a FDR adjusted p-value\u2009<\u20090.05 were finally considered. We observed high levels of methylation for CREBBP in the majority of tumor samples, however, similar patterns were manifest in normal tissue controls, hence CREBBP hypermethyation does not appear to be a feature of oncogenesis in these samples.For the supervised analysis of the probes, CpG sites were selected by applying an ANOVA test to identify statistically significant CpG positions that were differentially methylated among the compared groups (\u0394\u03b2\u2009>\u20090.2). Selected CpG sites were later clustered based on the Manhattan distances aggregated by ward\u2019s linkage. Finally, the genes corresponding to the selected CpGs were used to perform a Gene Set Enrichment Analysis (GSEA) with curated gene sets in the Molecular Signatures Databasehttp://github.com/vqv/ggbiplot). For each group we plotted a normal data ellipse with size defined as a normal probability equal to 0.68. Unsupervised hierarchical clustering was performed with the stats::hclust function using the 75 probes with the highest variance in methylation beta values.For principal component analysis, we used the R function prcomp to calculate the Singular Value Decomposition of the beta value matrix after removing the CpGs without methylation information. We plotted the first two principal components which contain most variation by using the ggbiplot R package algorithmThe Kaplan-Meier method was used to analyze survival outcomes by the log-rank Mantel-Cox test, with hazard ratio and two-sided 95% confidence intervals calculated using the Mantel_Haenszel test . Overall survival data from time of first surgery for each anaplastic meningioma within gene-expression defined subgroups C1 and C2 was collected and used to plot a Kaplan-Meier survival curve.TP53 p.R248Q missense mutation and a homozygous truncating variant in the mismatch repair gene MSH6 (p.L1330Vfs*9). Despite the latter finding, mutational signatures analysis was dominated by signature 1, with no evidence of signatures typically associated with defects in homologous recombination, mismatch repair or POLE activity . The copy number profile is most consistent with this tumor having first undergone haploidization of its genome, with the exception of chromosomes 7, 19 and 20, followed by whole genome duplication had a hypermutator phenotype, with 27,332 point mutations and LOH across nearly its entire genome (Supplementary Fig.\u00a0Supplementary File 1Supplementary Dataset 1"} +{"text": "RESULTS/ANTICIPATED RESULTS: We studied a heterogeneous cohort of patients with coronary artery disease (57%), peripheral artery disease (30%), carotid artery stenosis (7%), cerebral artery aneurysm (3%), and stroke (3%) on clopidogrel therapy for secondary prevention of thromboembolic events. The mean TPR was 205\u00b149 PRU (range: 61\u2013304), with a prevalence of 28% patients with high TPR (PRU\u2265230). No significant clinical differences were found between the non-high TPR and high-TPR groups (p>0.05). However, multivariable logistic regression analysis showed that both diabetes mellitus and proton-pump inhibitors were independently correlated with high TPR (p<0.05) after adjusting for other clinical variables. DISCUSSION/SIGNIFICANCE OF IMPACT: These results provide new insight into the importance of clinical characteristics on platelet reactivity in this Caribbean population. Further studies are warranted to determine whether important clopidogrel pharmacogenes are related with platelet function in Hispanics, as well as the role of TPR in guiding antiplatelet therapy and predicting future adverse cardiovascular events in this population.OBJECTIVES/SPECIFIC AIMS: To determine the association between clinical characteristics and platelet reactivity in Hispanic patients on clopidogrel therapy. METHODS/STUDY POPULATION: A cross-sectional pilot study was performed in 58 Puerto Rican patients diagnosed with any type of vascular disease and actively receiving a maintenance dose of clopidogrel for at least 7 days. The study population was divided into 2 groups: Group I with non-high on-treatment platelet reactivity (TPR); Group II with high TPR. To determine the platelet function, P2Y12 reaction units (PRU) were obtained by VerifyNow"} +{"text": "Using data from 46,029,364 Medicare beneficiaries included in the 2015 Current Beneficiary Survey (MCBS), we examined the relationship between dual sensory impairment (DSI) \u2013 concurrent vision impairment (VI) and hearing impairment (HI) \u2013 and accompaniment to physician visits. Analyses examined reasons for accompaniment and self-reported sensory impairment was categorized as: no sensory impairment (89%), hearing impairment (HI) only (5%), vision impairment only (4%), and DSI (1%). There was no difference in odds of accompaniment among HI compared to those without sensory impairment ; however, VI and DSI were associated with accompaniment: . Our study further demonstrates that older adults with sensory impairment are accompanied to physician visits more often than those without sensory impairment, and transportation is the most frequently reported reason for accompaniment among adults with VI and communication for those with HI."} +{"text": "Mouse models of hypercholesterolaemia have been used to identify arterial proteins involved in atherosclerosis. As the liver is extremely sensitive to dyslipidemia, one might expect major changes in the abundance of liver proteins in these models even before atherosclerosis develops. Lipid levels were measured and a proteomic approach was used to quantify proteins in the livers of mice with an elevated low-density lipoprotein (LDL) and the presence of lipoprotein(a) [Lp(a)] but no atherosclerosis.P<0.05). A bioinformatic analysis of the 24 proteins showed the major effect was that of an enhanced antioxidant and lipid efflux response with significant increases in antioxidant and lipid metabolism proteins . Interestingly, human liver cells treated with Lp(a) showed significant increases in Gpx1 and Prdx6 but not Sod1 or Park7. The livers of Lp(a) mice showed an increased triglyceride but reduced phospholipid and oxidised lipid content. Two-dimensional gel electrophoresis and mass spectrometry analysis identified 24 liver proteins with significantly increased abundance in Lp(a) mice ( The presence of human LDL and Lp(a) in mice promotes an enhanced flux of lipids into the liver which elicits an antioxidant and lipid export response before the onset of atherosclerosis. The antioxidant response can be reproduced in human liver cells treated with Lp(a). Mouse models of hypercholesterolaemia are widely used to study atherosclerosis. The two most widely used models are the apolipoprotein E deficient (ApoE-/-) mouse with eleThe hypercholesterolaemic mouse models have been subject to transcriptomic and proteomic approaches to investigate changes in arterial genes and proteins related to atherosclerosis development \u201310. A trThe liver has a central role in lipid metabolism and it synthesizes many of the proteins implicated in atherosclerosis. It is extremely sensitive to altered lipid flux driven by dyslipidemias and prolmus musculus) used here were the same wildtype C57BL/6 mice and Lp(a) transgenic mice used previously to investigate arterial protein expression [The mice on a Superose 6HR 10/30 column from GE Healthcare Bio-Sciences . Separated fractions were measured for cholesterol and triglycerides by enzymatic assay . et al. [The livers from 12 Lp(a) transgenic mice and 12 wildtype mice were rinsed in phosphate-buffered saline (PBS) and stored at -80\u00b0C. Lipids were extracted from 500 mg biopsies of each liver following the method by Bartels et al. . Tissue et al. was usedThe remaining frozen liver sections from 12 Lp(a) transgenic and 12 wildtype mice were homogenized in Tri-reagent containing 2% of a protease inhibitor cocktail (Roche) on ice. Proteins were precipitated with isopropanol and subsequently washed with ethanol and contaminants were removed using a 2D cleanup kit . The liver protein extracts were used for 2D PAGE proteomic analysis using the Mini-PROTEAN 2D electrophoresis system . Three replicate gels were run for each mouse. Following electrophoresis, all gels were stained with colloidal Coomassie brilliant blue and scanned with a calibrated densitometer . The 2D PAGE images were analyzed with the ImageMaster 2D platinum software and spots exhibiting a statistical difference (p<0.05) were excised for identification by mass spectrometry. Western blots were used to validate the comparative 2D proteomic analysis for the antioxidant enzymes. Following 2D PAGE, liver proteins were transferred onto nitrocellulose membrane using the PROTEAN Trans-Blot transfer system (Bio-Rad). Membranes were incubated overnight with primary polyclonal antibodies against Gpx1 (ab22604), Prdx6 (ab59543), and Sod1 (ab13498). Membranes were then incubated with HRP-conjugated anti-rabbit IgG or HRP-conjugated anti-goat IgG for 2 hours at room temperature. For the Park7 protein, liver proteins were separated by SDS PAGE and transferred onto nitrocellulose membrane and the membrane was probed with an anti-Park7 antibody (ab18257). Membranes were incubated in ECL reagent and exposed under standard sensitivity on a LAS-3000 luminescent image analyzer . Park7 protein levels were quantified by densitometry using Image Studio Lite after normalisation to actin.Lp(a) was isolated from human plasma as previously described using a combination of density ultracentrifugation and size exclusion chromatography . LDL was\u03bcg/mL amphotericin B, 100 U/mL penicillin, and 100 \u03bcg/mL streptomycin (Life Technologies) at 37\u00b0C in a humidified environment with 5% CO2. HepG2 cells were seeded at 5\u00d7105 cells/mL. 24 hours after seeding, cells were treated with 5 \u03bcg/ml of either purified Lp(a) or LDL for 6 hours at 37\u00b0C. Cell lysates were harvested and 40 \u03bcg of cell lysate protein was subject to western blot analysis with anti-Gpx1 , anti-Prdx6 (ab16947 Abcam), and anti-SOD (ab13498 Abcam) antibodies. An anti-actin antibody (Novus NB100-74340) was used as a loading control. Protein levels were quantified by densitometry using Image Studio Lite after normalisation to actin.Human hepatocellular carcinoma (HepG2) cells were maintained in Advanced Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum , 2 mM L-glutamine, 0.25 t-test was used to test for significant differences in mean hepatic lipid levels, the mean normalized spot volumes obtained from 2D PAGE analysis of the Lp(a) transgenic versus wildtype mice, the mean Park7/actin ratio, and the mean protein band intensity from lipoprotein-treated HepG2 cells. A difference with P<0.05 was considered significant. The AmiGO search tool was used to screen the gene ontology (GO) database to annotate the functional relevance of proteins identified as being significantly different in relative abundance between Lp(a) and wildtype mice. Protein network analysis was performed by uploading the list of proteins to the online STRING database tool (http://string-db.org) [Statistical analyses were performed using GraphPad Prism . A nonpaired Student's-db.org) and seleThe plasma lipid levels of the Lp(a) mice used for this study have been previously reported and show elevated cholesterol, triglyceride, and phospholipid levels as a result of elevated LDL and HDL cholesterol and the presence of Lp(a) . SeparatThe livers of Lp(a) transgenic mice showed similar cholesterol but signRepresentative 2D PAGE images from the livers of Lp(a) and wildtype mice on which the proteomic analysis was based are shown in Figures Western blot analysis of 2D PAGE gels for the antioxidant enzymes Sod1, Gpx1, and Prdx6 confirme in vitro on antioxidant proteins in the livers of Lp(a) transgenic mice, human HepG2 liver cells were treated with the two purified lipoproteins. Treatment with Lp(a) significantly increased the expression of GPx1 and Prdx6, but not Sod1 or Park7 or LDL could elicit a direct effect Figures . Treatme Figures .This study aimed to establish the changes in hepatic proteins in the Lp(a) mouse model of hypercholesterolemia. This model constitutes a milder form of hypercholesterolaemia than the apoE-/- or LDLR-/- mice , 2 and p\u03b2-oxidation enzymes (Acaa2 and Echs1). An increase in \u03b2-oxidation might also underlie the increase in the Uqcrc1 and Atp5h subunits of the electron transport chain. This response was in contrast to that seen in the arteries of the Lp(a) mice where there was significant decrease in Fabp4 and electron transport chain proteins [Despite significantly elevated plasma cholesterol levels , the Lp [\u03b2-oxidation thus preventing their breakdown to aldehydes. In contrast, the arteries of the Lp(a) mice accumulate TBARS and only show an increased abundance of one antioxidant enzyme, Prdx4, [An increase in es (ROS) . An incres (ROS) . Intereses (ROS) . The thres (ROS) , whereases (ROS) , 27. Intes (ROS) . Upregules (ROS) \u201331. The , Prdx4, which, uOur results contrast with studies of apoE-/- mice that showed a decrease in antioxidant capacity with significant decreases in antioxidant enzymes in both liver mitochondria and arteWith respect to lipid metabolism, the proteomic data also indicated an increased abundance of proteins involved in lipid export. Compared to the livers of wildtype mice, the Lp(a) mice had significant increases in both Apoa1 and Apoa4 . ApoA1 fLp(a) is primarily cleared by the liver via a nuWe have performed a proteomic analysis on the livers of human Lp(a) transgenic mice and shown an increased abundance in antioxidant and lipid export proteins. Our findings suggest that the initial response to mild hyperlipidaemia is to invoke protective mechanisms that reduce the accumulation of lipid and its oxidised products. This contrasts with mouse models of a similar age with more extreme hyperlipidaemia that have lost this protection mechanism and display atherosclerosis."} +{"text": "Ran and Stx, and we hypothesized that chromosomal proximity of these two genes across metazoans could be related to a regulatory logic or a functional linkage. We studied, for the first time, Ran expression during amphioxus development and reported its presence in the neural vesicle, mouth, gill slits and gut corresponding to body regions involved in active cell division.Ran (ras-related nuclear protein) is a small GTPase belonging to the RAS superfamily that is specialized in nuclear trafficking. Through different accessory proteins, Ran plays key roles in several processes including nuclear import-export, mitotic progression and spindle assembly. Consequently, Ran dysfunction has been linked to several human pathologies. This work illustrates the high degree of amino acid conservation of Ran orthologues across evolution, reflected in its conserved role in nuclear trafficking. Moreover, we studied the evolutionary scenario of the pre-metazoan genetic linkage between Xenopus laevis egg extracts has demonstrated the involvement of Ran pathway in mitotic progression antibody as a mitosis-specific marker in amphioxus to understand if there is a positive correlation of its expression pattern with cellular mitotic activity during development . PHH3 isRan is an atypical small GTPase that is localized exclusively in nucleus and usually linked to macromolecular transport through nuclear pores. This protein plays a pivotal role in diverse mitotic steps in mammalian cells, like spindle formation and chromosome separation , and hasS. salar show partial conservation of this amino acid stretch (DVQ), suggesting a possible diversification during evolution.Since Ran is so important for aspects of eukaryotic cell behaviour, we inquired into its molecular history broadly in Eukarya domain . Except Ran and Stx, two unrelated genes, belongs to a list of duplets unchanged across over 600 million years during evolution for 2 h at 4\u00b0C. The labeled specimens were rinsed in seven 20-min changes of PBT, mounted in 80% glycerol in PBS, and imaged by Axio Imager with ApoTome (Zeiss).S1 FigLatimeria chalumnae (Ran2) has been excluded from the tree for its divergence. Values at the branches indicate replicates obtained using the ML estimation method.The Maximum Likelihood (ML) tree indicates the existence in gnathostomes of Ran1 (orange box) and Ran2 (red box). The Ran of (PDF)Click here for additional data file.S2 FigIn particular, the Maximum Likelihood (ML) tree evidences a common evolutionary origin for Stx1 (blue box), Stx2 (light blue box) and Stx3 (violet box). Values at the branches indicate replicates obtained using the Maximum Likelihood estimation method.(PDF)Click here for additional data file.S3 FigA color code has been used to represent orthologue genes.(PDF)Click here for additional data file.S4 Fig(PDF)Click here for additional data file.S5 FigCollection of two series of embryos showing slightly different PHH3 immunolocalization signals (red). Scale bars: 60 \u03bcm. Embryos orientation: anterior to the left, dorsal to the top.(PDF)Click here for additional data file.S1 File(PDF)Click here for additional data file.S2 File(PDF)Click here for additional data file."} +{"text": "The present data profiles a large scale transcriptome changes in seed tissues (embryo and endosperm) during maturation in dormant and non-dormant genotypes of hexaploid wheat. Seed dormancy is an adaptive trait that has a significant influence on the incidence of preharvest sprouting, which is referred to as the germination of grains on the spike prior to harvest, in wheat. Given that preharvest sprouting causes a substantial yield and quality losses, elucidation of the molecular features that regulate seed dormancy has a paramount significance in the development of preharvest sprouting resistant wheat cultivars. The data presented here was produced from total RNA/mRNA samples isolated from developing seeds of dormant and non-dormant wheat genotypes using the Affymetrix GeneChip Wheat Genome Array. The raw and normalized formats of these data are available in Gene Expression Ominbus (GEO), NCBI's gene expression data repository, with accession number GSE83077. Reproducibility of the transcriptomic data from the independent replicates of each sample was confirmed by scatter plot expression analysis with the squared Pearson correlation coefficient (R2) . The raw and normalized formats of these data are available in Gene Expression Ominbus (GEO), NCBI's gene expression data repository (ent (R2) , Fig.\u00a04.22.1Plant of the dormant wheat genotype AC Domain and the non-dormant genotype RL4452 2.2Total RNA was extracted from both embryo and endosperm tissues. The total RNA from the embryos was isolated using RNeasy Plant Mini Kit while the total RNA from endosperm tissue was isolated as described previously 2.3The total RNA or mRNA samples were used for cDNA synthesis. After purification, biotinylated cRNA samples were prepared using the GeneChip IVT Labelling Kit and the GeneChip Sample Cleanup Module (Affymetrix). Assessment of the quality of the labelled cRNAs was undertaken using an Agilent 2000 Bioanalyzer. Subsequent to fragmentation, labelled cRNA samples were hybridized for 16 hr at 45\u00a0\u00b0C on GeneChip Wheat Genome Array. Washing and staining of the GeneChips were performed in the Affymetrix Fluidics Station 450. Afterwards, the GeneChips were scanned using an Affymetrix Scanner 3000.2.4http://harvest.ucr.edu/) 2) (Using the Affymetrix Microarray Suite (MAS5) statistical algorithm, the number of probesets with \u2018present\u2019 detection was determined. Subsequently, the raw data was normalized using Robust Multi-array Average (RMA) methodology. HarvEST WheatChip (edu/) 2) . As a reedu/) 2) , Fig.\u00a02 edu/) 2) , Fig.\u00a04"} +{"text": "Thousands of loci across the genome have been identified for specific diseases in genome-wide association studies (GWAS), yet very few are associated with lifespan itself. We hypothesized that specific biological pathways transcend individual diseases and affect health and lifespan more broadly. Using the published results for the most recent GWAS for 10 key age-related diseases we identified 22 loci with a strong genetic association with at least three of the diseases. These multi-trait aging loci include known genes affecting multiple diverse health end points, such as CDKN2A/B (9p21.3) and APOE. There are also novel multi-trait genes including SH2B3 and CASC8, likely involved in hallmark pathways of aging biology, including telomere shortening and inflammation. Several of these loci involve trade-offs between chronic disease risk and cancer."} +{"text": "TAZ knockdown significantly reduced expression of SOX2 at both mRNA and protein levels, whereas its ectopic overexpression markedly increased its abundance in HNSCC cells. Moreover, reintroduction of ectopic SOX2 abolished, at least in part, the reduced tumorsphere formation and tumorigenicity in vivo induced by TAZ knockdown. Mechanistically, transcriptional complex formed by TAZ and TEAD4 was recruited to two binding sites in SOX2 promoter, which in turn facilitated transcription of SOX2 in HNSCC cells. In addition, the abundance of TAZ and SOX2 was positively correlated in HNSCC clinical samples, and both upregulations of TAZ and SOX2 associated with the worst survival. Taken together, our data reveal a previously unknown mechanistic linkage between TAZ and SOX2 and identify SOX2 as a direct downstream target of TAZ in modulating CSCs self-renewal and maintenance in HNSCC. These findings suggest that targeting TAZ-SOX2 axis might be a promising therapeutic strategy for HNSCC.The Hippo-TAZ signaling has emerged as a fundamental regulator underlying cancer stem cells (CSCs) stemness which intricately associates with local recurrence and metastatic spreading in head neck squamous cell carcinoma (HNSCC). However, the precise downstream targets of TAZ responsible for HNSCC CSCs maintenance remain largely underexplored. Here, we identified Sex determining region Y box 2 (SOX2) as a putative downstream target of TAZ to promote CSCs maintenance and tumorigenicity in HNSCC. Both TAZ and SOX2 were significantly enriched in CSCs subpopulation (CD44 The overwhelming majority of these tumors are squamous cell carcinoma (SCC) and account for approximately 3.8% of all cancer cases and 3.6% of cancer-related deaths worldwide2. Several etiological factors like tobacco smoking, alcohol consumption, chewing of betel quid and human papillomavirus (HPV) infection have been identified for HNSCC. Current clinical management of HNSCC including ablative surgery, radiotherapy and chemotherapy has yielded remarkable progress in the past decades. However, the long-term survival rate remains dismal and only 40\u201350% of patients survive for more than 5 years since initial diagnosis3. Molecular targeted therapies to treat advanced, recurrent or metastatic HNSCC are lacking or with limited success in selected patients4. These challenges largely hinge on our incomplete understanding about molecular tumorigenesis of HNSCC as well as its genetic and biological heterogeneities. Thus, in-depth investigations of molecular pathways underlying HNSCC pathogenesis will lead to novel effective therapeutics and optimal treatment guiding, ultimately improving patients survival and quality of life5.Head and neck cancer comprises a heterogeneous group of malignancies that arise in the mucosal surfaces of the upper aerodigestive tract including oral and nasal cavity, pharynx and larynx as well as paranasal sinus6. These aggressive events have been revealed to be intricately associated with a unique cell subpopulation in bulk cancer, termed cancer stem cells (CSCs) or tumor-initiating cells (TICs) with potent self-renewal and tumor-seeding properties. These cells sustain tumor overgrowth and drive recurrence and metastatic dissemination8. Moreover, these unique characteristics have enabled CSCs as attractive and yet challenging therapeutic targets as evidenced by the facts that targeting key regulators or molecular pathways responsible for CSCs properties has yielded remarkable anti-cancer effects with promising translational potentials9. Previous pioneering work has documented several surface or functional markers for HNSCC CSCs including CD44, CD133, Bmi1, SOX2 and ALDH112. Particularly, SOX2 (Sex determining region Y box 2), the key transcriptional factor preferentially expressed in embryonic and adult stem cells, has been demonstrated as an essential marker and regulator underlying these unique properties of CSCs from diverse cancer origins15. Previous reports have documented some essential clues to support SOX2 as a key CSC regulator in HNSCC19. However, the detailed mechanisms concerning how SOX2 itself is regulated in HNSCC remain incompletely known.Locoregional recurrence, cervical metastatic spread and chemoresistance largely account for therapeutic failure and cancer-related death in HNSCC20. Dysregulation of Hippo signaling essentially contributes to cancer initiation, outgrowth, metastatic dissemination and therapeutic resistance. Upon Hippo inactivation, two downstream effectors transcriptional coactivator PDZ-binding motif (TAZ/WWTR1) and yes-associated protein (YAP) are translocated into nucleus whereby they drive transcription of target genes mainly by forming complexes with TEA domain DNA-binding family of transcription factors (TEADs)21. Importantly, we and others have reported that elevated TAZ promotes self-renewal and tumor-seeding potentials of CSCs and also confers CSCs-like properties on differentiated non-CSCs in diverse cancer contexts24. However, the accurate downstream targets responsible for TAZ in HNSCC CSCs self-renewal and maintenance remain underexplored yet.The Hippo signaling pathway has increasingly been recognized as a pivotal and indispensable mediator in tissue homeostasis, regeneration and tumorigenesisHere, we sought to determine whether SOX2 was a novel downstream target of TAZ underlying CSCs properties in HNSCC. Our findings by integrating cellular experiments in vitro, tumor-forming assay in xenograft animal model as well as bioinformatics data mining provide evidence that TAZ enhances CSCs self-renewal and maintenance by direct transcriptional activation of SOX2 in HNSCC.25. However, the accurate downstream targets responsible for TAZ underlying HNSCC CSCs properties remain largely underexplored. To address this, we set out to perform initial screen of dozens of putative CSCs regulators such as CD44, Nanog, SOX2, Bmi1 and ALDH1 as potential candidates via shRNA-mediated TAZ knockdown approach in vitro (data not shown). Among these candidates screened, SOX2 attracted our attentions due to the following reasons: its consistent and significant reduction upon TAZ silencing in both Cal27 and Fadu cells, its well-established roles in CSCs in squamous cell carcinoma as well as its intricate link with Hippo pathway in other biological contexts26. TAZ knockdown mediated by shRNA lentiviral constructs remarkably reduced both mRNA and protein abundance of SOX2, as well as CTGF (a well-known downstream target of TAZ) in vitro to predict potential binding sites of TEAD4 in human SOX2 promoter region (2000 bp upstream of TSS and 150\u2009bp downstream of TSS) and identified eight potential binding sites and demonstrated its robust self-renewal and tumor-initiating potentials both in vitro and in vivo23. As shown in Fig. S+CD133+ CSCs subpopulation yielded much more and larger tumorsphere in vivo as compared to CD44\u2212CD133\u2212 subpopulation (non-CSCs). Then we utilized two independent siRNAs targeting human SOX2 to knockdown its expression and subsequently monitored the resulting phenotypic changes in vitro with stable TAZ knockdown, SOX2 overexpression and TAZ knockdown plus SOX2 overexpression were subcutaneously inoculated into both blanks of NOD/SCID mice. Consistently, as shown in Fig. 6 cells within 4 weeks. Data from volume during tumor growth, final volume and weight of tumor samples indicated that TAZ and SOX2 facilitated tumor overgrowth in vivo. Notably, enforced SOX2 overexpression was capable to partially abrogate the anti-growth effects resulted from TAZ knockdown. Then, tumor samples derived from Fadu cells with TAZ or/and SOX2 manipulations were subjected to further analyses. As shown in Fig. To recapitulate these in vitro findings in vivo, we utilized the limited dilution and tumorigenic assay. Various amounts of cells experiments in Cal27 and Fadu cells and found endogenous TEAD4 in complex immunoprecipitated by TAZ specific antibody without TEAD4 binding ability failed to increase luciferase activities of SOX2 promoter , while no significant correlations between TAZ/SOX2 and other clinicopathological parameters were identified. In addition, significant correlation between TAZ and SOX2 expression was found in these samples examined in breast cancer and OSCC23. To extend these findings and bridge the gap between TAZ and CSCs stemness in HNSCC, we screened dozes of known CSCs regulators and found that SOX2 might be downstream target of TAZ in HNSCC as evidenced by its downregulation or upregulation followed by TAZ knockdown or overexpression in vitro. Consistent with previous findings regarding SOX2 in HNSCC stemness31, our data further confirmed the pivotal roles of SOX2 in CSCs maintenance in HNSCC as evidenced by impaired abilities of tumorsphere formation, reduced expression of CSCs markers and compromised tumor initiation in vivo following SOX2 knockdown. Additionally, both TAZ and SOX2 were significantly enriched in CSCs subpopulation and were significantly downregulated during serum-induced tumorsphere differentiation. More importantly, experimental findings derived from limiting dilution and tumorigenic assay in vivo further revealed the pivotal roles of TAZ and SOX2 required for HNSCC tumor initiation and overgrowth. Collectively, these findings provide compelling evidence to support the essential roles of TAZ and SOX2 in CSCs self-renewal and maintenance in HNSCC.Our previous results have showed that TAZ mediates self-renewal and maintenance of CSC in OSCC as evidenced by its enrichment in CSC subpopulation, impaired tumorsphere formation and reduced CSC percentage upon TAZ knockdown, as well as positive associations between TAZ expression and tumor aggressiveness in OSCC samples32. Disruption of TAZ-TEADs binding blocked the major effects mediated by TAZ including oncogenic transformation, pro-proliferation and EMT in cancer33. Here, we identified TEAD4 as the key partner with TAZ to regulate SOX2 expression in HNSCC through IP assay, which was also consistent with its oncogenic functions and aberrant upregulation in cancer34. Subsequently, our results from luciferase reporter assays involving SOX2 promoter and its mutant as well as ChIP assays further confirmed the direct binding between TAZ/TEAD4 and SOX2 promoter in HNSCC. Noticeably, TAED binding sites in SOX2 promoter identified here were different from those found in other reports whereby two putative YAP/TEAD binding sites were found at upstream (\u22123759) and downstream (\u2009+\u20095313) of SOX2 transcription start site (TSS) in murine osteosarcoma cells35. We reasoned that it\u2019s conceivable that binding sites for transcriptional factors vary in diverse cell types and biological settings. More importantly, reintroduction of SOX2 into cells with TAZ stable knockdown abrogated, at least in part, the impaired proliferation, migration and tumorsphere formation, and in vivo tumor initiation and growth induced by TAZ depletion. Interestingly, several previous reports revealed that YAP1 transcriptionally activated SOX2 expression through a physical interaction with OCT4 to facilitate self-renewal of stem-like cells in lung cancer28. SOX2 antagonized Hippo pathway leading to exaggerated YAP functions to maintain stemness in osteosarcoma15. However, our data failed to support TAZ as a potential, direct downstream target of SOX2, which was consistent with previous report wherein TAZ was unaffected in SOX2-depleted osteoprogenitors26. Taken together, these in vitro and in vivo findings offer experimental support that TAZ-TEAD4 complex activates SOX2 transcription by direct binding with its promoter, which in turn modulates CSCs stemness in HNSCC. These findings identified another upstream regulator of SOX2 and added another layer of complexity to the transcriptional regulatory network governing CSCs unique characteristics.Previous studies have established that TAZ primarily functions as a transcriptional coactivator in complex with TEADs to mediate downstream transcriptional events30. However, the expression pattern of SOX2 and its prognostic significance in HNSCC remain inconsistent and contradictory. Some reports found that SOX2 was frequently amplified in HNSCC and its upregulation associated with favorable survival, while others documented the opposite results40. In our patient cohort, we failed to identify positive or negative association between SOX2 expression and patient survival (data not shown). These discrepancies might be due to sample size, tumor heterogeneity as well as methods of patient stratification. Nonetheless, our results revealed that patients with TAZhigh/SOX2high had the worst survival ratios as compared to other patient subgroups. Of course, further studies are needed to resolve the inconsistency regarding the prognostic value of SOX2 in multiple, large and independent HNSCC cohorts. Noticeably, we developed a prognostic score based on TAZ/SOX2-correlated genes via bioinformatics and statistical approaches and found that this score had potent power to stratify HNSCC patients into subgroups with favorable or inferior survival. This suggests that the regulatory network modulated by TAZ and SOX2 in HNSCC might not only intricately contribute to tumorigenesis, but also might be exploited as a novel prognostic biomarker with translational promise in the clinic.The prognostic value of TAZ has been established in several human cancer including OSCC and HNSCCIn conclusion, our data reveal a previously unknown mechanistic linkage between TAZ and SOX2 and identify SOX2 as a direct downstream target of TAZ in modulating CSCs self-renewal and maintenance in HNSCC. These findings suggest a novel therapeutic approach to target TAZ/TEAD4-SOX2 signaling axis in HNSCC.2. Mycoplasma detection was routinely performed during the whole course of this study. All regents were purchased from Sigma-Aldrich unless otherwise stated.A panel of HNSCC cell lines including Cal27, Fadu, HN6 and human embryonic kidney 293\u2009T (HEK293T) was used here. Cal27, Fadu and HEK293T were obtained from American Type Culture Collection and authenticated by short tandem repeat profiling at regular intervals. HN6 was a generous gift from Prof. Wantao Chen (Shanghai Jiaotong University). All cancerous cells lines were maintained in DMEM/F12 (Gibco) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin at 37\u2009\u00b0C in 5% CO23. The TAZ mutant plasmids (TAZ4SA and TAZ4SA+S51A) were kindly gifted from Prof. Kunliang Guan41. The human full-length SOX2 or TEAD4 cDNA with 3\u2009\u00d7\u2009Flag was subcloned into lentiviral plasmid pLenti CMV/Puro and then verified by direct sequencing. Lentiviral particles were prepared by transiently co-transfecting HEK293T cells with individual lentiviral constructs and controls together with packaging and envelope plasmids (pCMV-VSV-G and pCMV-\u03948.2) using the calcium-phosphate method. These viral supernatants were filtered, concentrated and stored until use. For transient transfection assay with siRNA or plasmids, cells were harvested at 48\u2009h for further experiments. To gain stable clones after infections with shRNA or overexpression lentiviral vectors, cells were selected with puromycin for at least one week.Two independent sequences of siRNA or shRNA targeting human SOX2 and TEAD4 mRNA (detailed sequences were listed in Table TM RT-PCR kit (Takara) was used for qRT-PCR reactions, as we described previously42. Endogenous 18\u2009S RNA or GAPDH was used for data normalization. All qPCR primers used were listed in Table Total RNA of tissue specimens or cells was extracted with Trizol reagent (Invitrogen) and then subjected to transcription into cDNA by PrimeScript\u2122 RT Master Mix (Takara) according to the manufacturer\u2019s instructions. PrimeScript+ cells were identified under fluorescent microscopy, photographed and counted via ImageJ software. Cell invasion was assessed using transwell chambers with 8-\u03bcm pore size (Corning) with pre-coated Matrigel (BD Pharmingen) as we described previously43.Cell proliferation and viability were assessed by absorbance using CCK-8 cell viability assay and BrdU incorporation assay according to manufacturer\u2019 instructions. BrdU23. Briefly, for apoptosis detection, cells were trypsinized, dissociated into single cell suspension, then assayed with Annexin V: PE Apoptosis Detection Kit (BD Bioscience) for flow cytometry. For FACS, single cell suspension was incubated with CD44 and CD133/1 and two subpopulations of CD44+CD133+ and CD44\u2212CD133\u2212 was separated when corresponding immunoglobulins was used for blank control. All data were collected and analyzed by BD FACSuite software.Flow cytometry for cell apoptosis and fluorescence-activated cell sorting were similar as we reported previously23. GAPDH was used as a loading control. For co-IP assay, cells were lysed by the Western & IP Lysate Buffer supplemented with 1% protease inhibitor (Roche) on ice for >15\u2009min. Cell lysate was centrifuged under 4\u2009\u00b0C for 10\u2009min at 12,000\u2009rpm, and the supernatant was incubated with primary antibodies and protein A/G agarose beads (Thermo Fisher Scientific) with rotating at 4\u2009\u00b0C overnight. The pellet was washed at least three times with IP lysate buffer on ice and then subjected to western blotting analysis. All antibodies used were listed in Table Western blot analyses were routine performed as described previouslyThe promoter sequence (2065\u2009bp) upstream of the transcriptional start site of human SOX2 was subcloned into a luciferase reporter plasmid and verified with direct sequencing. Two putative binding sites between TEAD4 and SOX2 were individually mutated using QuikChange\u00ae Lightning Site-Directed Mutagenesis Kits (Stratagene) and verified by direct sequencing. The 293\u2009T cells were transiently transfected with pGL-SOX2, TAZ or TEAD4 plasmids and phRL-CMV plasmid (Promega) using lipofectamine 2000 (Invitrogen). Cells were harvested 24\u2009h after transfection and assayed for Firefly and Renilla luciferase activity using the Dual-Luciferase reporter system (Promega). Data were presented as the ratios of Firefly to Renilla luciferase activity.TM Chromatin Immunoprecipitation Kit (Millipore) according to the manufacturer\u2019s instructions. Isolated and purified chromatin was incubated overnight with antibodies for TEAD4 (Abcam), Flag (Sigma), RNA polymerase (Millipore), or normal mouse IgG (Millipore). RNA polymerase was utilized as positive control and normal mouse IgG was used as negative control. The PCR or qPCR primers for detecting CTGF, SOX2 binding site 1 and 2 were listed in the Table For ChIP assay, Cal27 cells that had been transfected with Flag-TAZ for 48\u2009h were harvested and lysed using the EZ-ChIP4/ml) were cultured in serum-free DMEM/F-12 supplemented with B27, 20\u2009ng/ml EGF (BD Bioscience) and 10\u2009ng/ml bFGF (BD Bioscience) and grown in ultra-low-attachment plates (Corning) at routine conditions for 7\u201310 days. For in vitro serial passages, these tumorsphere were harvested and further dissociated into single cells by 0.1% trypsin and gentle pipette, and then filtered, re-plated to form secondary sphere in aforementioned medium. The tumorsphere with diameter larger than 50\u2009\u03bcm was counted.The disassociated single cells in 200\u2009\u03bcL media (100\u2009\u03bcL PBS plus 100\u2009\u03bcL PBS Matrigel) and then subcutaneously injected into both flanks of each animal . Tumor initiation and growth were monitored and recorded every three days. Tumor volume was calculated with the formula: volume\u2009=\u2009a\u2009\u00d7\u2009b2/2. Tumor weight was weighed and recorded after animals sacrificed. Tissue samples were fixed in formalin or maintained in liquid nitrogen for further analyses.All animal models involved in this experiment were in accordance with the institutional animal welfare guidelines and protocols were approved by Institutional Animal Care and Use Committee of Nanjing Medical University. Six-week-old female NOD/SCID mice were purchased from Model Animal Research Center of Nanjing Medical University and maintained in the specific pathologic-free animal facility. Cancer cells were trypsinized, counted and diluted with equal number were included in each staining run. Immunoreactivity was semi-quantitatively evaluated according to staining intensity and distribution using the immunoreactive score which was calculated as intensity score\u2009\u00d7\u2009proportion score as we previously reportedhttps://cancergenome.nih.gov/) and Gene Expression Omnibus (GEO) Database (GSE23036 and GSE65858)46. The mRNA expression levels of TAZ and SOX2 HNSCC were log2-transformed and compared using the Pearson correlation test.The original data concerning the expression of TAZ and SOX2 in HNSCC were retrieved from publicly available databases The Cancer Genome Atlas using TCGA-HNSCC dataset. The overlapped genes were considered as potential candidates for TAZ/SOX2-correlated gene signature, which were subjected to univariate regression assay to identify candidates which were significantly associated with overall survival in TCGA-HNSCC datasets. Then, these candidates were further filtered using Robust likelihood-based modeling for 1000 times via R environment with Rbsurv package and multivariate Cox regression analysis with top statistical significance48. A risk score formula based on the expression level and coefficient of these selected candidates was generated and its optimal cut-off point was selected at the maximal sensitivity and specificity by receiver operating characteristics (ROC) curve. Subsequently, the prognostic values of this score were validated in two publically available HNSCC cohorts (GSE41613 and GSE42743)49.The positively or negatively correlated genes of TAZ and SOX2 in HNSCC were initially downloaded from cBioPortal and P\u2009<\u20090.01 (**).All quantitative data in the present study were shown as mean\u2009\u00b1\u2009SD of three independent experiments unless otherwise stated. All data were analyzed using GraphPad Prism 7.0 or SPSS 22.0 software with Student\u2019s Supplementary TablesSupplementary Figure legendsSupplementary Fig S1Supplementary Fig S2Supplementary Fig S3Supplementary Fig S4Supplementary Fig S5Conflict of interest statement"} +{"text": "Using miDGE, we screened over 73 different PV genomes for the ability to code for miRNAs. Our results show that most PVs are unlikely to code for miRNAs and we conclusively demonstrate a lack of PV miRNA expression in cancers associated with infections of several high risk HPVs. However, we identified five different high-confidence or highly probable miRNAs encoded by four different PVs ). Extensive in vitro assays confirm the validity of these miRNAs in cell culture and two FcPV1 miRNAs are further confirmed to be expressed in vivo in a natural host. We show that miRNAs from two PVs (HPV41 & FcPV1) are able to regulate viral transcripts corresponding to the early region of the PV genome. Combined, these findings identify the first canonical PV miRNAs and support that miRNAs of either host or viral origin are important regulators of the PV life cycle.MicroRNAs (miRNAs) are small RNAs that regulate diverse biological processes including multiple aspects of the host-pathogen interface. Consequently, miRNAs are commonly encoded by viruses that undergo long-term persistent infection. Papillomaviruses (PVs) are capable of undergoing persistent infection, but as yet, no widely-accepted PV-encoded miRNAs have been described. The incomplete understanding of PV-encoded miRNAs is due in part to lack of tractable laboratory models for most PV types. To overcome this, we have developed Papillomaviruses (PVs) are causative agents of cancer. Currently, there is an incomplete understanding as to why only some infections lead to cancer. Developing a better comparative evolutionary understanding of PV gene products and their regulation is key to comprehending the life cycle of these pathogens. An emerging concept of viral gene regulation is that many persistent viruses will utilize small regulatory RNAs called miRNAs to optimize host and viral gene expression. Yet despite obvious interest, there have been no credible reports identifying canonical PV-encoded miRNAs. Here we develop new broadly applicable methodology to identify miRNAs from organisms lacking a laboratory culture system. We identify the first examples of bona fide canonical PV miRNAs and provide evidence supporting both host and viral miRNA-mediated regulation as relevant to control of PV gene expression. These findings resolve the issue of PV miRNAs and further the notion of miRNA importance to persistent virus infection. Papillomaviruses (PVs) comprise a large family of circular double-stranded DNA viruses. Numerous PV genomes have been described including over 200 human PV (HPV) types. A minority of these are known as carcinogenic agents \u20133, howevMicroRNAs (miRNAs) are small regulatory RNAs that are an emerging class of viral gene products found in select virus families \u20137. miRNAOver 300 viral encoded miRNAs have been described, all from viruses able to undergo long term persistent infection ,5,7,30. One barrier to discovery of PV miRNAs is the dearth of facile fully-infectious laboratory systems. There are experimental systems established for a few PV types (approximately < 5) , but tecmiRNA Discovery by forced Genomic Expression (miDGE). miDGE relies on generating a library of numerous overlapping genomic segments of DNA from a particular organism or locus and subcloning them behind a heterologous RNA polymerase (RNP) II promoter . JMRV is a gamma-2 herpesvirus with genomic sequence similar to the highly related Rhesus Rhadinovirus (RRV). When we initiated these studies, it was not yet known if JMRV encoded miRNAs, although this would be expected since numerous precursor miRNAs (pre-miRNAs) had already been identified in RRV, which shares high sequence similarity with JMRV . Indeed, by JMRV . Thus, Jin vitro. As shown in de-novo prediction of pre-miRNAs with the miRDeep2 algorithm with 10 \u03bcl \u03b2-mercaptoethanol) and homogenized. The lysate was centrifuged at maximum speed in a microcentrifuge for 3 min and transferred to a fresh microcentrifuge tube. Ethanol (100%) was added to the cleared lysate to bring the final concentration up to 60% ethanol. Next, the samples were applied to an RNeasy (Qiagen) mini-spin column to purify the total RNA according to the manufacturer's instructions, except that after the final wash step, the samples were stored at approximately 4\u00b0 C for several days while still on the column. The final elution steps were conducted with one volume of nuclease-free water and then repeated with one volume of nuclease-free Tris-EDTA (TE), pH 7. Pooled small RNA-seq libraries were prepared from RNA harvested from either foot lesions or pectoral muscle from animals CF180/09 and CF229/12. Equal volumes of total RNA were combined and ethanol precipitated. Recovered RNA was dissolved in nuclease free water (Ambion) and quantitated with a NanoDrop spectrophotometer (ThermoFisher). Libraries were prepared using 180 ng of pooled RNA with the NEBNext Multiplex Small RNA Library Prep Set for Illumina according to the manufacturer\u2019s instructions with the addition of an additional round of indexing PCR to compensate for low input RNA. Libraries were quantitated with the Qubit dsDNA BR assay (ThermoFisher), QC checked with the Bioanalyzer High Sensitivity assay (Agilent Technologies), combined with other barcoded libraries, and sequenced on a single lane of a SR50 run on a HiSeq 4000 (Illumina) by the Genomic Sequencing and Analysis Facility at UT Austin. 106,251,321 and 122,129,107 reads were obtained for the foot and pectoral samples respectively (SRA accession: SRP133175). Small RNA reads were pre-processed by trimming the adaptor sequences and removing trimmed sequences shorter than 18 nucleotides with Cutadapt (version 1.4.2) [Total RNA was extracted from both leg lesions and pectoral muscle (control) tissue from two chaffinches with proliferative leg skin lesions that were PCR-positive for FcPV1 DNA. The affected chaffinches were found dead by members of the public and submitted for post-mortem examination to a national scanning surveillance scheme for wild bird disease in Great Britain . Reads wn 1.4.2) to the rn 1.4.2) and FcPVThe Cancer Genome Atlas cervical cancer RNA-seq datasets were retrieved from the NCI Genomic Data Commons. From the large RNA-seq data sets, sequenced coverage was calculated for the HPV reference sequence with the greatest number of alignments. Tumors with > = 50% coverage for an HPV were used for subsequent analysis . Aligned BAM files were converted to miRDeep2 format and the miRDeep2 pipeline was run with default parameters without miRNA annotations. BEDTools was used to assign the de novo miRDeep2 identified miRNAs to miRBase release 21 annotations.Small RNA data sets associated with NCBI GEO project GSE42380 were retrieved from the NCBI SRA. The SRA files were converted to colorspace FASTQ format using the SRA Toolkit. Adapter sequences were trimmed from reads using Cutadapt (version 1.4.2) . The triS1 Figi.e., only reads matching the proper miRNA sequences are shown).Each panel depicts small RNA-seq coverage in material from JMRV-infected fibroblasts (upper plots) or 293T cells transfected with our miDGE fragment library (lower plot in each panel). Gray bars under the plots indicate the location of pre-miRNA hairpins. Thicker sections denote the location of previously annotated mature miRNA sequences. Coverage is shown in a strand-specific manner for each miRNA and precursor ((TIF)Click here for additional data file.S2 Fig(A) and the two positive control polyomaviruses miRNAs contained in our library (B). Grey bars underneath the plots indicate the location of pre-miRNA hairpin sequences, with thicker boxes denoting the location of mature miRNA products. Coverage is shown in a strand-specific manner for each miRNA and precursor .Plots show small RNA-seq read coverage from HEK293T cells transfected with PV miDGE libraries for the 5 novel papillomavirus miRNAs identified in this study (TIF)Click here for additional data file.S3 Fig(A) or 18 (B). Panel C shows coverage of the suggest HPV type 18 precursor by reads from our JMRV miDGE library transfection, demonstrating that coverage of these sequences is nonspecific. Open bars and boxes underneath the plots in (A) indicate the location of suggested precursor and mature miRNAs, respectively. Coverage in (A) is shown in a strand-specific manner for each miRNA and precursor . For the HPV type 18 candidate shown in (B) and (C), mature miRNAs (indicated by thick block arrows) were proposed to derive from both strands of the precursor (thinner block arrow). Therefore, plots in (B) and (C) show coverage across both strands of the viral genome.Plots show small RNA-seq read coverage from HEK293T cells transfected with PV miDGE libraries for 8 previously purported miRNAs in HPV types 6, 16, 38, and 45 (TIF)Click here for additional data file.S4 FigA diagram of the FcPV1 genomic organization is provided at the top showing the positions of the known open reading frames (ORFs). Below is a detailed view of the 3' untranslated region following the L1 ORF and the predicted base pairing with the let-7a-5p microRNA. Vertical bars \"|\" indicate predicted base pairing and \":\" indicates predicted wobble pairing. The let-7a sequence is from the closest relative with available genomic sequence, the zebra finch.(TIF)Click here for additional data file.S1 TableThe table shows counts of small RNA reads from JMRV-infected fibroblasts (\u201cinfection\u201d) or HEK293T cells transfected with our miDGE library (\u201cmiDGE\u201d) mapped to the indicated precursor or mature JMRV miRNA regions.(DOCX)Click here for additional data file.S2 TableColumn coverage provides the percentage of the viral genome that was covered by our miDGE libraries, according to high-throughput sequencing of the expression library. The column labeled acronym denotes our internal label for individual genomes .(DOCX)Click here for additional data file.S3 TableReference PV genomic sequences were downloaded from PAVE . Each se(DOCX)Click here for additional data file.S1 DatasetThis dataset contains DNA-seq coverage data across viral genomes contained in our miDGE PV libraries in bedgraph format.(BG)Click here for additional data file.S2 Dataset(A) and the designation of known or experimentally confirmed novel pre-miRNAs matching the miRDeep2 predictions (B). \u2018l.c.\u2019 in column B indicates likely false positive predictions which can be eliminated by filtering reads for low complexity filters prior to performing the miRDeep2 analysis.This dataset shows an overview of all predictions made by miRDeep2 for our JMRV and PV libraries. Columns C to N provide miRDeep2 prediction data in the pipeline\u2019s standard tabular output format. Columns A and B indicate the miDGE library for which the predictions were made (XLSX)Click here for additional data file.S3 DatasetThis dataset shows miRDeep2\u2019s primary output (in PDF format) of read coverage along hairpin structures for all predictions made by the pipeline for our JMRV miDGE data. Provisional IDs for the individual pre-miRNAs are given as assigned by the pipeline, see (PDF)Click here for additional data file.S4 DatasetThis dataset shows miRDeep2\u2019s primary output (in PDF format) of read coverage along hairpin structures for all predictions made by the pipeline for our PV miDGE data (including the two PyV positive controls). Provisional IDs for the individual pre-miRNAs are given as assigned by the pipeline, see (PDF)Click here for additional data file.S5 DatasetThis dataset contains information on where each PV genomic plasmid used in this study was obtained.(XLSX)Click here for additional data file.S6 DatasetThis dataset contains information about the sequences of reporters and probes used in this study.(XLSX)Click here for additional data file.S7 DatasetThis dataset contains primary data used in re-analysis of Qian et al data , quantif(XLSX)Click here for additional data file.S8 DatasetThis dataset includes the full uncropped scans of northern blots used in (PDF)Click here for additional data file.S9 DatasetThis dataset includes the coordinates of human miRBase release 21 microRNA seed matches (positions 2\u20138 inclusive) in all PAVE annotated papillomavirus genomes.(XLSX)Click here for additional data file."} +{"text": "In older adults, unintentional weight loss (UWL) is associated with poor outcomes, but its pathophysiology remains poorly understood. We sought to identify potential biomarkers of UWL using targeted metabolomics, including 8 conventional metabolites, 45 acylcarnitines, and 15 amino acids. We identified individuals from the Cardiovascular Health Study All Stars with UWL (n=40) or weight stability from Years 9 to 11. Participants had WS through Year 8. UWL was defined as experiencing >6% weight loss from Years 9 to 11 and self-reporting that loss as unintentional. Mean plasma metabolite concentrations measured in Year 9 were compared between individuals with UWL or WS between Years 9 and 11. The strongest signals in metabolomic differences between individuals going on to experience UWL versus WS were observed among the branched-chain amino acids, valine and isoleucine/leucine ; lactate ; histidine ; the medium-chain acylcarnitine octenoyl carnitine (C8:1) ; and long-chain acylcarnitine myristoyl carnitine (C14) . These findings suggest altered branched-chain amino acid and fatty acid metabolism and increased oxidative stress and inflammation may be evident before individuals undergo UWL. Further investigation of these pathways may reveal novel preventive or treatment strategies for UWL."} +{"text": "The WBRTMel trial is a multinational, open-label, phase III randomised controlled trial comparing whole brain radiotherapy (WBRT) to observation following local treatment of one to three melanoma brain metastases with surgery and/or stereotactic irradiation. The primary trial endpoint was to determine the effect of adding WBRT to local treatment on distant intracranial control, and the secondary endpoints were neurocognitive function, quality of life (QoL), performance status, overall survival, death from intracranial causes, death from melanoma and cost-effectiveness.The objective of this update is to outline and publish the pre-determined statistical analysis plan (SAP) before the database lock and the start of analysis.The SAP describes basic analysis principles, methods for dealing with a range of commonly encountered data analysis issues and the specific statistical procedures for analysing efficacy and safety outcomes. The SAP was approved after closure of recruitment and before completion of patient follow-up. It outlines the planned primary analyses and a range of subgroup and sensitivity analyses regarding the clinical and QoL outcomes. Health economic outcomes are not included in this plan but will be analysed separately. The SAP will be adhered to for the final data analysis of this trial to avoid analysis bias arising from knowledge of the data.The resulting SAP is consistent with best practice and will allow open and transparent reporting.We have developed a SAP for the WBRTMel trial which will be followed to ensure high-quality standards of internal validity to minimise analysis bias.ACTRN12607000512426. Registered on 9 October 2007. ClinicalTrials.gov, NCT01503827. Registered on 4 January 2012. Trial group reference numbers ANZMTG 01.07, TROG 08.05.ANZ Clinical Trials Registry, Brain metastases are a common cause of death in patients with melanoma. The use of whole brain radiotherapy (WBRT) after excision and/or stereotactic radiosurgery (SRS) for melanoma brain metastases is variable because of the lack of high-quality evidence to guide practice. The trial\u2019s rationale, feasibility and protocol were previously published \u20133. In brThe statistical analyses will assess the clinical endpoints and safety of WBRT compared with the observation following excision and/or SRS.This trial aims to improve the treatment of brain metastases for patients with stage IV melanoma by using WBRT to improve disease control and maintain cognitive performance and QoL.The primary endpoint of the study is defined as distant intracranial failure within 12\u2009months of randomisation. Distant intracranial failure is evaluated through magnetic resonance imaging (MRI) assessment, as reported by participating institutions, and is defined as new lesions appearing 1\u2009cm or more from previous index metastases, coded as a binary outcome of success or failure.B \u2013 HVLTF) \u00f7 HVLTB, where indexes B\u2009=\u2009baseline and F\u2009=\u2009a pre-specified follow-up time point ), even those not analysed, will be reported stratified by treatment arm.We have developed a statistical analysis plan (SAP) for the ANZMTG 01.07 WBRTMel study. This plan will be followed to ensure high-quality standards of internal validity to minimise analysis bias. The SAP was approved and signed off by the Trial Management Committee on 1 October 2018. Following data integrity checks of the primary and secondary 12 months outcomes, the statistical analysis specified in the SAP was started January 2019.A list of proposed tables and a list of proposed figures are displayed in this section. Reporting of the trial results is based on the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) guideline for reporting clinical trial results .(1)\u00a0Patient characteristics stratified by treatment arm(2)\u00a0Medical history by treatment arm(3)\u00a0Distant disease at baseline by treatment arm(4)\u00a0Lesions summary by treatment arm(5)\u00a0Physical examination by treatment arm(6)\u00a0Systemic therapy and steroid use over treatment by treatment arm(7)\u00a0Treatment compliance by treatment arm(8)\u00a0Primary and secondary distant intracranial failure by treatment arm(9)\u00a0Key secondary outcome(10)\u00a0The EuroQol visual analogue scale (EQ VAS)(11)\u00a0Deterioration in NCF over time by treatment arm(12)\u00a0Subgroup analysis (replicate Tables\u00a0(8) and (10)\u00a0for each subgroup as defined in Subgroups\u00a0section)(13)\u00a0Tertiary outcomes (replicate Tables (1) (8)\u00a0and (9), stratified by Observation vs. HA-WBRT vs. WBRT)(14)\u00a0AE and SAE summary by treatment arm.Trial flowchart Cumulative incidence function of distant intracranial failure (competing with death) stratified by treatment armCumulative incidence function of local intracranial failure stratified by treatment armCumulative incidence function overall intracranial failure (competing with death) stratified by treatment armCumulative incidence function of melanoma death (competing with other cause of death) stratified by treatment armCumulative incidence function deterioration in cognitive performance (competing with death) stratified by treatment armCumulative incidence function of melanoma death (competing with non-melanoma death) stratified by treatment armCumulative incidence function of intracranial death stratified by treatment armCumulative incidence function of deterioration in role function (competing with death)Cumulative incidence function of global QoL (competing with death)Cumulative incidence function of drowsiness (competing with death)Cumulative incidence function of communication difficulties (competing with death)Cumulative incidence function of motor dysfunction (competing with death)Cumulative incidence function of social function items/domains (competing with death)Evolution of Hopkins Verbal Learning Test-Revised, Delayed Recall (HVLT-R DR) and Controlled Oral Word Association Test stratified by treatment armEvolution of HVLT-R, Total Recall and Delayed Recognition stratified by treatment armEvolution of Controlled Oral Word Association Test stratified by treatment armEvolution of Trail Making Test Parts A and B stratified by treatment armEvolution of Stroop Color and Word Test (Adult Version) stratified by treatment armEvolution of Digit Span (Forward and Backward) stratified by treatment armKaplan\u2013Meier overall survival curves by treatment armCumulative incidence of local intracranial failure stratified by Observation vs. HA-WBRT vs. WBRTCumulative incidence of time to distant intracranial failure stratified by Observation vs. HA-WBRT vs. WBRTKaplan\u2013Meier overall survival stratified by Observation vs. HA-WBRT vs. WBRTEvolution of HVLT-R DR stratified by Observation vs. HA-WBRT vs. WBRTEvolution of HVLT-R, Total Recall and Delayed Recognition stratified by Observation vs. HA-WBRT vs. WBRTKaplan\u2013Meier time to global cognitive deterioration \u2013 HVLT-RLikert scale for EQ-5D-5\u2009L dimensions/itemsLikert scale for EORTC QLQ-C30 dimensions/itemsLikert scale for EORTC QLQ-BN20 dimensions/items"} +{"text": "The genomic landscape of head and neck cancer has been reported through The Cancer Genome Atlas project. We attempt to determine if high-risk human papillomavirus (HPV) or frequently mutated genes are correlated with survival in an oral cancer cohort.Patient demographic data along with data from final pathology was collected. Tumor DNA was analyzed using a custom Illumina targeted sequencing panel. Five high-risk HPV types were tested by qPCR. Statistical analyses were used to identify associations between patient outcome and mutational status.p\u2009=\u20090.03).High-risk HPV types were identified in 7% of cases; HPV status was not associated with survival. Mutations were identified in TP53, TERT promoter, & PIK3CA. Mutations in TP53 were significantly associated with poorer overall survival on multi-variate analysis (Mutations in TP53 were associated with poor patient survival. Expanding our sample size may identify further predictors of outcome to direct customized cancer care. Oral cavity squamous cell carcinoma (OSCC) is a significant public health problem worldwide, with over 350,000 new cases diagnosed yearly and greater than 150,000 annual deaths .Exposure to tobacco and alcohol represent conventional risk factors for the development of OSCC. However, oral human papillomavirus (HPV) infection has recently been discovered to be an additional risk factor for a significant number of head and neck cancers, particularly those arising in the oropharynx. The most common subtypes identified by The Cancer Genome Atlas (TCGA) were HPV16 84%), HPV33 (11%), HPV35 (4%) and HPV56 (1%) %, HPV33 , howeverTP53 mutations and 3p arm deletion events were previously found to be predictors of tumor recurrence and overall patient survival within the TCGA cohort had very poor coverage and was removed from downstream analyses. Similarly, a single normal sample [395] also had very poor coverage and was removed. The matched tumor sample was subsequently treated as tumor only.Additional file 3: Figure S3. Comparison of mutations by smoking status. CASP8 and TERT promoter mutations occurred more frequently in patients with less than 10 pack year smoking history.Additional file 4: Table S1. Summary of TP53 mutations.Additional file 5: Table S2. Univariate Analysis for Overall Survival.Additional file 6: Table S3. Univariate Analysis for Disease-free Survival."} +{"text": "Andean potato mottle virus from Peru (isolate Lm) was obtained. Comparison of its RNA1 and RNA2 sequences with variants of this virus isolated in Brazil revealed RNA1 and RNA2 nucleotide identities of 81 to 83% and 70 to 71%, respectively.A complete coding sequence of the type strain of Andean potato mottle virus from Peru (isolate Lm) was obtained. Comparison of its RNA1 and RNA2 sequences with variants of this virus isolated in Brazil revealed RNA1 and RNA2 nucleotide identities of 81 to 83% and 70 to 71%, respectively.A complete coding sequence of the type strain of Andean potato mottle virus , and one infected Tetragonia expansa (Aizoaceae) , complete APMoV sequences are lacking.In 1972, virus isolate Lm was obtained from a potato plant showing leaf mottle growing at La Molina in Lima, Peru. It constitutes the type strain of zoaceae) . Later, zoaceae) , 5 and pzoaceae) was repozoaceae) and partSequencing methods were as previously described \u201311. Brie\u2013Comovirus sequences. Amino acid sequence comparisons with Brazilian APMoV isolates showed that Lm\u2019s combined CPs had 68% amino acid identity with these sequences, whereas their Pro-Pol had 94% amino acid identity. In sequence comparisons between Lm and partial Chinese APMoV isolates\u2019 CP and Pol, amino acid identities were 74% (CP) and 96% (Pol). The nearest comoviruses were Squash mosaic virus (42% CP identity) and Turnip ringspot virus (45% Pro-Pol identity). The latest Secoviridae International Committee on Taxonomy of Viruses (ICTV) report (https://gd.eppo.int/taxon/AVBO00), such studies are needed to ascertain the plant health status of its divergent isolates.Lm\u2019s RNA1 sequence had 81 to 83% nucleotide identities with partial RNA1 sequences of Brazilian APMoV isolates , 8, wher) report states tMN176101 (RNA1) and MN176102 (RNA2). In the SRA, raw data were deposited under BioSample number SAMN12259772 and BioProject number PRJNA491634.The APMoV genome sequences were deposited in GenBank under the accession numbers"} +{"text": "Hemophilia A (HA) is an X-linked recessive disorder caused by clotting factor VIII (FVIII) deficiency. There is limited data on the use of replacement therapy in cardiac surgery. Since no international guideline for anticoagulation in such patient exists, careful thought should be taken to design an individualized anticoagulation strategy.We report a 54-year-old male with severe HA with FVIII activity of 0.8% when he was first diagnosed, who underwent successful mitral valve repair and coronary artery bypass graft with FVIII replacement perioperatively.Transthoracic echocardiography and coronary angiography confirmed the HA patient with the diagnosis of severe mitral valve regurgitation and left anterior descending artery stenosis.Before surgery, a bolus of 1000 IU FVIII was injected, which obtained an FVIII of 80%. After induction, a 3750 IU bolus of FVIII was injected and subsequent FVIII level reached 135%. Mitral valve repair and coronary artery bypass graft with FVIII replacement were performed. After the surgery, a repeat FVIII activity level was 50.6%. The 400 mL of autologous blood and 700 mL of cardiopulmonary bypass (CPB) machine blood was returned to the patient as well as 4 units of fresh frozen plasma with an additional bolus of 1000 IU FVIII. 100\u200amg aspirin per day alone was given after surgery.The patient recovered uneventfully and 1-year follow-up showed no complications.The anticoagulant or antiplatelet regimen of HA patient following surgery should be individualized based on the evaluation of the risk factors for bleeding and thrombosis and the lowest FVIII activity ever recorded after FVIII replacement therapy. HA is classified according to plasma procoagulant levels into mild (5%\u201340%), moderate (1%\u20135%) and severe (<1%). There is limited data on the use of replacement therapy in those requiring cardiopulmonary bypass (CPB) for complicated cardiac surgery. Here we report a 54-year-old male with severe HA who underwent successful mitral valve repair (MVP) and coronary artery bypass graft (CABG) with FVIII replacement perioperatively. Since no international guideline for anticoagulation in such patient exists, careful thought should be taken to design an individualized anticoagulation strategy.2A 54-year-old male (body weight 75 Kg) was admitted to our hospital with acute chest tightness, asthma and shortness of breath after unexpectedly sliding down 3 steps 1 week ago. The patient had a history of HA with FVIII activity of 0.8% when he was first diagnosed. After the diagnosis, he received replacement therapy (13.3\u200aIU/kg of FVIII each time) twice a week, which maintained an FVIII activity ranging from 2% to 80%. And he underwent bilateral knee replacement 5 years ago due to spontaneous hemorrhage. He has a 30-year history of smoking and drinking without withdrawal. Physical examination revealed a grade IV/VI holo-systolic murmur heard best at the apex and wet rales in both lungs. Transthoracic echocardiography (TTE) showed a posterior leaflet prolapse of mitral valve (P2 mainly) with moderate to severe regurgitation due to rupture of tendinae, and a left ventricular diastolic diameter of 56\u200amm and an ejection fraction of 70% of FVIII was injected, which obtained an FVIII of 80%. After induction, a 3750 IU (50\u200aIU/kg) bolus of FVIII was injected and subsequent FVIII level reached 135%. A tranexamic acid infusion of 1\u200ag/h (lasting 10\u200ah) was then started before incision. An intravenous introducer sheath (9F) was placed in the right internal jugular vein, and 400\u200amL of autologous blood was harvested. A dose of heparin (375\u200aIU/kg) for CPB resulted in an activated clotting time (ACT) \u2265300\u200aseconds before the left internal mammary artery (LIMA) was severed. Surgery was performed with LIMA anastomosed to LAD after cardioplegic arrest, mitral valvuloplasty involving a triangular resection of the posterior leaflet, creation of an artificial chordae (Gortex CV-4) and insertion of an annuloplasty ring . The CPB lasted around 95 minutes, and a transesophageal echocardiography examination was performed that aspirin should be used 81 to 325\u200amg/d before and 6\u200ahours after CABG surgery, and should be used lifelong to reduce bridge obstruction and adverse cardiac events. Many patients with coronary artery disease are accompanied by valvular disease at the same time, so in recent years more and more patients receive simultaneous CABG and valve surgery. As for which antithrombotic therapy should be used after MVP, there is no clear consensus at the moment. The European guidelines recommend taking vitamin K antagonists for anticoagulation therapy for 3 months post mitral valvuloplasty, but the application varies from country to country. American surgeons chose antiplatelet therapy (grade 2C), 64% of British surgeons chose warfarin for 6 months after MVP, while 54% of surgeons use aspirin long-term. Currently in China, most cardiac surgeons choose warfarin therapy for 3 months after MVP. Nevertheless, in the handful of HA patients underwent CABG plus valve surgery reports out there, a range of coagulation management plans were used, including heparin alone, warfarin alone, aspirin plus warfarin for 30 days, and not using anything at all. Aspirin is well tolerated in HA patients. Considering that our patient had severe HA and a history of bilateral knee replacement due to spontaneous bleeding with a low CHA2DS2-VASc score for chance of stroke and we only used a single LIMA bridge and an annuloplasty ring for MVP, his lowest FVIII activity after 1000 IU FVIII replacement therapy every other day was 8.1%, thus we came to the conclusion that 100\u200amg aspirin alone per day was suitable postoperatively. One year follow-up showed that the patient was well and had neither bleeding nor thrombosis event.There are few reports of patients with HA undergoing cardiac surgery, only 7 cases of CABG combined with valve surgery have been reported in English literature,4In conclusion, we believe that patients with severe HA can undergo simultaneous CABG and valve surgery if a suitable coagulation management strategy is enacted. Following surgery, the individual anticoagulant or antiplatelet regimen can be selected based on evaluation of the risk factors for bleeding and thrombosis and the lowest FVIII activity ever recorded after FVIII replacement therapy.Conceptualization: Yanyan Yang.Data curation: Davies Henry.Investigation: Chengcheng Li.Methodology: Chengcheng Li.Validation: Yanyan Yang.Writing \u2013 original draft: Hongfei Xu.Writing \u2013 review & editing: Haige Zhao."} +{"text": "Development of new drugs with other mechanisms of action (MOA) in chronic lymphocytic leukemia (CLL) than those in clinical practice are highly warranted in spite of the recent clinical progress, especially compounds targeting tumor-specific molecules. Therapeutics interfering with signaling pathways controlling growth and survival of leukemic cells and bypass resistance to cytotoxic drugs is of special interest and tyroThe receptor tyrosine kinase (RTK) ROR1 is normally expressed during embryogenesis but repressed in most adult tissues. We and others have shown high expression of ROR1 in CLL cells includinA small molecule, KAN0439834 (535\u2009Da), targeting the TK domain of ROR1 was developed from a library of 110.000 compounds using fresh CLL cells (disease related cell phonotype selection procedure) specifically inhibiting phosphorylation of the TK domain .+ cells) from patients with non-progressive or progressive CLL (as defined by IWCLL criteria), as well as of patients with fludarabine resistant disease with or without del(17p) were analyzed for KAN0439834 induced cytotoxicity. A dose-dependent cytotoxicity was observed, which also included fludarabine resistant CLL cells with and without del(17p) (EC50 250\u2009nM) was >60 folds at EC50. Apoptosis was confirmed by Annexin V/PI staining . KAN0439834 induced significant apoptosis of CLL B cells but not of normal T cells Fig.\u00a0. There wNext, we compared apoptosis induced by KAN0439834 with that of ibrutinib, idelalisib, and venetoclax (ABT-199) on CLL cells from different compartments of the same patients. KAN0439834 and venetoclax induced a similar high degree of apoptosis of CLL cells from the three compartments while ibrutinib and idelalisib only killed CLL cells obtained from lymph nodes, but to a lower degree than KAN0439834 and venetoclax Fig.\u00a0. KAN0439KAN0439834 induced a dose-dependent dephosphorylation of ROR1 in CLL cells (WB) Fig.\u00a0, which w\u2212 stromal cells) were co-cultured, HS-5 cells could partially prevent apoptosis of CLL cells at low concentrations of KAN0439834, while at higher concentrations the presence of stromal cells had no effect increased phosphorylation of ROR1 in a dose-dependent manner, but not SRC which might be activated by other kinases . KAN0439ect Fig.\u00a0. MoreoveROR1 and the co-receptor LRP6 may heterodimerize in CLL cells as part of signal transduction . KAN0439KAN0439834 bound also to a few other kinases (KINOMEscan) but at high concentrations (10\u2009\u00b5M) (50 for those inhibitors were >10\u2009\u00b5M using the same target (CLL cells). Importantly, dephosphorylation of ROR1 was not observed by inhibitors such as bemcitinib (R428) (AXL inhibitor), idelalisib or ibrutinib, all of which, however, (as expected) dephosphorylated their respective targets at a high dose of KAN0439834 but less pronounced at a low dose. A similar significant reduction of CD45+/CD19+/ROR1+ cells was also noted when CLL cells harboring del(17p) were transplanted but not as marked as for non-del(17p) CLL cells. A significant dose-dependent reduction of ROR1 expression was noted in both experiments, as well as reduction of phosphorylated ROR1. IHC analyses supported the results of flow cytometry of KAN0439834 \u2014a tyrosine kinase inhibitor. Novel drugs with other MOA are warranted to further improve the prognosis in CLL and related disorders. KAN0439834 may be a drug candidate also for other ROR1 expressing hematological and non-hematological tumors alone or in combination with other therapies.Supplementary Methods, Information and Figures"} +{"text": "ASNS (asparagine synthetase) that may affect substratebinding, and variants in driver genes including TP53, PIK3CA, FGFR2,ARID2, MLL3, MYC and ALK. Using the IntOGenplatform, we identified MAP kinase, cell cycle, actin cytoskeleton regulation,PI3K-Akt signaling and other pathways in cancer as affected in the samples. Thisdata is the first of its kind from the Pakistani population. The results of thisstudy can guide a better mechanistic understanding of HNSCC in the population,ultimately contributing new, rational therapeutic targets for the treatment ofthe disease.Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancertype globally and contributes significantly to burden of disease in South Asia.In Pakistan, HNSCC is among the most commonly diagnosed cancer in males andfemales. The increasing regional burden of HNSCC along with a unique set of riskfactors merited a deeper investigation of the disease at the genomic level.Whole exome sequencing of HNSCC samples and matched normal genomic DNA analysis(n=7) was performed. Significant somatic single nucleotide variants (SNVs) wereidentified and pathway analysis performed to determine frequently affectedsignaling pathways. We identified significant, novel recurrent mutations in Head and neck squamous cell carcinomas (HNSCC), which include tumours of the oralcavity, oropharynx, hypopharynx and larynx, are the sixth most common cancerworldwide with a global incidence of ~600,000 cases andbetel quid or paan along with its related products are additional risk factors inthis part of the world . Written informed consent to participate was obtained from allsubjects.2 was collectedand stored in RNAlater\u00ae solution (Thermo Fisher Scientific) at -80 \u00b0Ctill further processing. Formalin-fixed tumour tissue samples were assessed by ahistopathologist for tumour content and cellularity based on hematoxylin andeosin (H&E) staining. Seven tumour samples negative for HPV with at least70% cancer cells and 1 \u03bcg (50 ng/\u03bcl) of extracted DNA (both tumour as well asgenomic DNA) were utilized for whole exome sequencing.Fresh tumour tissue and matched blood were obtained from treatment-na\u00efve patientsundergoing surgical resection of HNSCC primary tumour at the Aga Khan UniversityHospital in Karachi, Pakistan. Patients with confirmed histological diagnosis ofHNSCC were included in this study. At the time of resection, fresh tumour tissueaway from the necrotic core measuring at least 0.5 cmGenomic and tumour DNA was extracted in-house using TRIzol\u00ae Reagent according to manufacturer\u2019s instructions. Tumour DNA was extracted from atleast 50 mg of tissue and genomic DNA was extracted from 3-5 mL of peripheralblood samples obtained before patients underwent surgical procedure. DNA yieldand quality was assessed both in-house using a NanoDrop 2000 spectrophotometer and by Macrogen Inc. usingPicoGreen\u00ae Assay .insitu hybridization (ISH) was performed on FFPE blocks usingGenPoint assay according to the manufacturer\u2019s instructions .Dako assay can detect HPV-DNA from 13 high-risk genotypes.Formalin-fixed paraffin embedded (FFPE) tumour blocks were retrieved and DNA wasextracted for assessing HPV status. PCR detection was performed using two setsof general HPV primers (GP5/GP6) . 1-2 \u03bcg of tumour andgenomic DNA was fragmented by nebulization. DNA libraries were prepared fromeach sample using TruSeq DNA Sample Prep Kit using the manufacturer\u2019s protocol. Unique molecular indices were used for each sample. Exomeenrichment was performed using the TruSeq Exome Enrichment kit .Paired-end sequencing was performed on Illumina HiSeq 2000 instrument. Each readwas of 100 bp size.The data sets supporting the results of this article are included within thearticle and its supplementary files. The raw sequencing data of those patientsthat consented to deposition of data in a public database have been deposited in NCBI\u2019s Sequence Read Archive and are accessible throughaccession number SRP083063.p-value of \u2264 0.05 wasapplied. The annotation ranked the SNVs for somatic driver mutations forspecific cancer tissue types, predicted protein functional impact, allelefrequencies from the 1000 Genomes Project and ESP6500 populations, and previouscancer association of the gene harbouring the variants. CHASM training set iscomposed of a positive class of driver mutations from the COSMIC database andVEST training set comprising a positive class of disease mutations from theHuman Gene Mutation Database 66 and a negative class of variants detected in theESP6500 population and 1000 Genomes Project cohort with an allele frequency of>1%. SNPeff (p<10\u20135 applied to predict benign anddeleterious variants (p\u22640.05) affected pathways in the cohort and genes withinidentified.Paired-end sequence reads from Illumina were mapped against UCSC Human Genome(hg) 19 using BWA .Local rThe homology model of human asparagine synthetase was constructed using crystalstructural coordinates of the enzyme from Escherichia coli (PDB id 1CT9). TheModeller program was usedFigureS1), along with their matched genomic DNA,were used for this study. The detailed demographics and clinical characteristicsof these patients are provided in Primary tumour samples from 7 treatment-na\u00efve HNSCC patients. Nonsense and splice sitevariants were also identified in all samples (Table S3).Paired-end whole exome sequencing (WES) of all seven HNSCC samples and matchedgenomic DNA was performed on Illumina HiSeq 2000 platform. Each read was of 100bp size. Additional details of the sequencing, including coverage and depth, aresummarized in re novel ; Table 3FGFR2 (Fibroblast Growth Factor Receptor 2),SETBP1 (SET Binding Protein 1), PIK3CA, IGF2BP3 (Insulin Like Growth Factor 2 MRNA BindingProtein 3), TP53 (Tumour Protein P53), PTPN11 and NF2(Neurofibromin 2) were identified. Significant missense mutations were alsoidentified in ASNS (Asparagine Synthetase(Glutamine-Hydrolyzing)) in four of the seven samples. Other genes thatexhibited recurrent mutations included the CLMN gene(5/7), CHEK2 (Checkpoint Kinase 2) (3/7), andDRD5 (Dopamine Receptor D5) and PAK2 (P21(RAC1) Activated Kinase 2) (2/7) (2/7) . These rARID2(AT-Rich Interaction Domain 2), ALK (Anaplastic LymphomaReceptor Tyrosine Kinase), MLL3 and MYC , were also identified ) and two genes involved in eukaryotic translation initiation[EIF4B (Eukaryotic Translation Initiation Factor 4B) andEIF4A3 (Eukaryotic Translation Initiation Factor 4A3)] wereseen in two of the seven samples (Table S3). Significant non-coding mutationswere filtered using CADD (Table S4). In the 3\u2019UTR region, mutationsin IGF1R (Insulin Like Growth Factor 1 Receptor) andERBB4 (Erb-B2 Receptor Tyrosine Kinase 4) were identifiedas significant. Another eukaryotic translation initiation factor,EIF2B4 (Eukaryotic Translation Initiation Factor 2B SubunitDelta), exhibited significant splice site variance. IntOGen pathway analysisrevealed that the MAP kinase pathway was the most significantly affected pathwayin all samples tested. In addition, cell cycle, actin cytoskeleton regulation,PI3K-Akt signaling and other pathways in cancer were among those significantlyenriched for exomic alterations in all samples are located in the vicinity (within 10 \u00c5 distance) of the glutaminebinding pocket . Previous studies have reported greater number ofmutations in HPV-negative as compared to HPV-positive HNSCC tumours . As a coTP53, which is associated with smoking-relateddisease, and the oncogene PIK3CA, at a mutation rate of 40-60% and6-8%, respectively and missense mutations have been previously identified exclusively in HNSCC recurrences gene in 4 out of 7 samples. TheseSNVs in ASNS have not previously been reported in the literature assignificant in HNSCC pathogenesis. The ASNS gene codes for aubiquitously expressed, ATP-dependent enzyme that converts aspartate and glutamineto asparagine and glutamate , an HNSCC clinicalsub-type, in the Indian population revealed frequently altered genes that arespecific to OSCC-GB and others that are also affected in HNSCC . AlteredNOTCH1 and HRAS were not identified inthis small cohort. However, given limited resources, it was deemed important toestablish preliminary data prior to a larger scale study. The approach of using asmaller discovery cohort followed by validation of identified mutations in a largercohort has been proposed and taken by others and reported in the literature (The small sample size is a limitation of this study, which may explain low frequencyof commonly mutated genes and why some of the commonly occurring HNSCC mutationssuch as ASNS and other genes in the Pakistanipopulation. It has been well established that a complex interplay of genetic andenvironmental factors results in varying risk of cancer development and treatmentoutcomes across different ethnicities and geographic regions (This is the first report describing the mutational spectrum of Pakistani HNSCCpatients. In addition to reporting known HNSCC mutations, we have identified novel,recurrent mutations in regions . Therefo"} +{"text": "Correction to: Thromb J (2017) 15:24https://doi.org/10.1186/s12959-017-0143-3Affiliation 3 should read \u201cUniversity of Sheffield, Sheffield, United Kingdom\u201d and Affiliations 6, 7, 8 and 9 were unnecessary duplicatesFGA followed by four mutations in FGB and two mutations in FGG, respectively.\u201dIn the abstract the sentence \u201cTen patients had mutations in FGA followed by three mutations in FGB and three mutations in FGG, respectively\u201d should be \u201cTen patients had mutations in In the Results section the following three sentences:Following the publication of this article , the aut\u201cIn FGA gene, eight mutations were identified as novel and the remaining two were reported mutations. Eight novel mutations include five missense, one nonsense and two frameshift mutations including homozygous and a compound heterozygous frameshift mutation. The two nonsense mutations in FGA are reported in literature. There is one more mutation with reported status in proband (C3). This patient had compound heterozygous mutation with frameshift as novel mutation and nonsense as reported. We identified three mutations in FGB including one novel missense mutation (C9) and two homozygous nonsense mutations reported in siblings. The FGG gene mutations are the rarest of all three fibrinogen genes. We detected three novel mutations including two similar nonsense mutations in siblings and one frameshift mutation in unrelated proband in different exons of FGG gene Table .\u201dShould be:4)There are a number of errors in Tables 5)Frameshift mutation (p.Gln282Thr fsx83*) and (p. Lys (AAA) 48Arg fs9*) are the novel compound heterozygous mutations which have manifested deletions along with frameshift defects\u201d should in fact read \u201cFrameshift mutations (p.Thr283Arg fs138*) and (p. Lys (AAA) 48Arg fs9*) are the novel compound heterozygous mutations which have manifested deletions along with frameshift defects.\u201cIn FGA gene, seven mutations were identified as novel and the remaining three were reported mutations. Seven novel mutations include five missense and two frameshift mutations including homozygous and a compound heterozygous frameshift mutation. The three nonsense mutations in FGA are reported in literature. There is one more mutation with reported status in proband (C3). This patient had compound heterozygous mutation with frameshift as novel mutation and nonsense as reported. We identified four mutations in FGB including one novel missense mutation (C9), two homozygous nonsense mutations reported in siblings and one frameshift mutationC12). The FGG gene mutations are the rarest of all three fibrinogen genes. We detected two novel similar nonsense mutations in siblings (Table 2. The FG"} +{"text": "Gata-1 mRNA. The major cellular processes affected by both Csde1 and Strap were ribosome function and cell cycle control.Erythropoiesis is regulated at many levels, including control of mRNA translation. Changing environmental conditions, such as hypoxia or the availability of nutrients and growth factors, require a rapid response enacted by the enhanced or repressed translation of existing transcripts. Cold shock domain protein e1 (Csde1/Unr) is an RNA-binding protein required for erythropoiesis and strongly upregulated in erythroblasts relative to other hematopoietic progenitors. The aim of this study is to identify the Csde1-containing protein complexes and investigate their role in post-transcriptional expression control of Csde1-bound transcripts. We show that Serine/Threonine kinase receptor-associated protein (Strap/Unrip), was the protein most strongly associated with Csde1 in erythroblasts. Strap is a WD40 protein involved in signaling and RNA splicing, but its role when associated with Csde1 is unknown. Reduced expression of Strap did not alter the pool of transcripts bound by Csde1. Instead, it altered the mRNA and/or protein expression of several Csde1-bound transcripts that encode for proteins essential for translational regulation during hypoxia, such as Hmbs, eIF4g3 and Pabpc4. Also affected by Strap knockdown were Vim, a Gata-1 target crucial for erythrocyte enucleation, and Elavl1, which stabilizes Ferritin and Transferrin receptor mRNA to control expression of the encoded proteins that are crucial to erythropoiesis . Cells were lysed in 850\u03bcl cold NT2, supplemented by 200U RNAse Out (EMD Bioscience), 400\u03bcM vanadyl ribonucleoside complexes and 20mM EDTA (EM Science), and incubated with the beads for 2 hours at 4\u00b0C. Beads were immobilized in a magnet rack, washed 5x with NT2 containing 0.3M NaCl, split into a protein and an RNA fraction. The protein fraction was eluted via boiling in 1x Laemmli buffer (Sigma-Aldrich) for 5 minutes. RNA fractions were purified using Trizol (Invitrogen), precipitated in isopropanol and washed in 75% ethanol.Cell lysates, SDS-PAGE, and Western blotting were performed as described previously . Biotaggprotocol , with thMEL cells were fractionated into cytoplasmic and nuclear components using a Cell Fractionation Kit\u2014Standard , or total MEL cell protein lysates were generated. Proteins were detected via SDS-PAGE and Western blotting as described . Antibodies used were directed against Strap , Csde1 , Stat5 , Lamin B1 and alpha Tubulin . Fluorescently labeled secondary antibodies for visualization with Odyssey were IRDye 680RD Donkey anti-Rabbit IgG and IRDye 800CW Donkey anti-Mouse IgG , or using the Pierce enhanced chemiluminescence (ECL) kit (Thermofisher). Silver staining was performed using a SilverQuest\u2122 Silver Staining Kit .t-test applying an artificial within groups variance of S0 = 0.8 was used [https://www.ebi.ac.uk/pride/).Eluted peptides were processed as described by . Sampleswas used . For allRNA-seq on Csde1-associated transcripts after Strap knockdown was performed by the Leiden Genome Technology Center , using library preparation following the template-switch protocol (Clontech), and Nextera tagmentation. Samples were split across three MiSeq (Illumina) lanes . RNA expression by total mRNA sequencing after Strap knockdown was performed by Novogene Co., LTD. on mRNA enriched on oligo(dT) beads. RNA was randomly fragmented, and processed with the NEB Next\u00ae Ultra\u2122 RNA Library Prep Kit using random hexamers. The library was sequenced using Illumina HiSeq 2500 . Sequence quality for both experiments was checked using Fastqc (Babraham Bioinformatics).https://www.ncbi.nlm.nih.gov/bioproject/).Spliced Transcripts Alignment to a Reference was useFor analysis of differential exon usage, we used the DEXSeq package ,39. DEXSRNA expression levels were normalized as reads per kilobase of transcript per million mapped reads (RPKM). In mass spectrometry, iBAQ values (as determined via MaxQuant) were normalized via a scaling factor calculated by dividing the sum of intensities from each sample by the intensity sum of a reference sample. A Spearman rank correlation coefficient was calculated between 10log(RPKM) and 10log(iBAQ).S1 Fig(A) A comparison between the base mean of pull downs from cells treated with shRNA against Strap and the pull down of cells treated with control virus (Sc Strap) (B) A comparison between the base mean of pull downs from MEL WT cells and the pull down of cells treated with control virus (Sc Strap). Colours indicate whether transcripts were specifically pulled down from cells expressing biotagged Csde1 versus BirA only at a FDR < 0.05. Red dots detected in parental MEL and in shRNA treated MEL; blue dots detected in previous study with parental MEL; green dots detected in shRNA treated MEL. Many transcripts are detected in both experiments (red dots). The transcripts that were not detected in the current study with virus transduced cells are detected at lower levels in the pull down of MEL WT cells, and transcripts that are detected in the current study but not in MEL WT are detected at higher levels upon virus transduction. An increased or decreased detection level does not affect the fold-change increase in cells that do or do not express biotaged Csde1. We assume that this is due to overall expression level.MEL cells expressing biotin ligase BirA with and without biotagged Csde1 (MEL WT) were treated with anti-Strap (Sh Strap) and control (Sc Strap) shRNA. They were then subjected to a protein-RNA pulldown on streptavidin beads followed by RNA sequencing. Shown are the reads from pull downs of cells that express biotagged Csde1. The base mean is the mean read count in counts per million (cpm) from 3 independent experiments. MEL WT cells were previously analysed ref. . Reads f(PDF)Click here for additional data file.S2 FigMEL cells expressing biotin ligase BirA with and without biotagged Csde1 were treated with anti-Strap and control (Sc) shRNA. They were then subjected to a protein-RNA pulldown followed by RNA sequencing. Cells expressing BirA without biotagged Csde1 represent pulldown background. An interaction term was used to model the effect of Strap knockdown on Csde1 transcript affinity. Significant transcripts are highlighted in red.(PDF)Click here for additional data file.S3 FigDepicted are both shRNA and replicate groups, indicating that the shRNA is responsible for the majority of variation between samples. PC2 (12%) is the result of minor batch effects.(PDF)Click here for additional data file.S4 Fig(A) Total cell lysates of MEL cells expressing BirA plus or minus biotagged Csde1 was used to pull down Csde1 using streptavidin beads. lysates were loaded on SDS-PAGE. Western blots were probed with anti-Csde1 and anti-Strap antibodies. The tagged Csde1 protein pulled down on streptavidin beads, has been extend with 23 amino acids (B) Western blot loaded with lysate fractions from parental MEL cells (WT), or CRISPR clones with bi-allelic deletions in Csde1 indicated as hypomorphic , or deleted and heterozygous deletion (HET KO). Lysates were fractionated into cytoplasmic C) and nuclear (N) extracts. Numbers identify specific CRISPR clones (see ref. see ref. Antibody(PDF)Click here for additional data file.S5 Fig(A) Venn diagram depicting the number of Csde1-bound transcripts (blue), and the number of differentially expressed transcripts detected at the transcript level (orange) or single exon level comparing MEL cells treated with Sc control shRNA or anti-Strap. (B) Examples of transcripts with alternative exon usage between MEL cells expressing Sc (blue line) or anti-Strap (red line) shRNA. Transcript names (short and full) and function are indicated, expression is in cpm on a 10log scale. Exons are numbered on the x-axes, which corresponds to the graphic representation of all exons (in grey) below, together with known transcript variants. The differentially expressed exon is pink, and indicated with a red arrow.(PDF)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S7 Table(XLSX)Click here for additional data file.S8 Table(XLSX)Click here for additional data file.S9 Table(XLSX)Click here for additional data file.S10 Table(XLSX)Click here for additional data file."} +{"text": "Mdx mice were randomly assigned into three groups: mdxS, the control group receiving intraperitoneal (i.p.) injections of saline solution (100\u03bcL); mdxP, positive control group receiving prednisolone (1mg/kg) by oral gavage; and mdxT, treated group receiving i.p. injections of tempol (100 mg/kg). C57BL/10 mice were also used as controls. Tempol treatment promoted gain in muscle strength and reduced myonecrosis and inflammatory response in the dystrophic diaphragm (DIA) and biceps brachii (BB) muscles. No evidence of Tempol's beneficial performance on angiogenesis in DIA and BB mdx muscles was found. The findings presented here show that Tempol treatment improves dystrophic phenotype, supporting its use as a potential therapeutic strategy in DMD.Considering potential Tempol effects on Duchenne muscular dystrophy (DMD) is the most common muscular dystrophy, whose genetic defect is identified in the X chromosome gene that encodes the intracellular protein dystrophin . DMD patmdx mice, the high intracellular calcium levels are directly related to the increase of oxidative stress and exacerbated inflammation octane mounting medium for fluorescence microscopy (Nikon Eclipse TS100). F4/80 staining was quantified using NIS-elements AR Advances Research software. For F4/80 immunofluorescence the percentage of total muscle area was calculated in each section studied (four or five sections per muscle). A blinded observer carried out the counts and measurements.mdx mice was determined by counting blood vessels with positive staining for CD31 , the pattern protocols were in agreement with to those reported by Verma , migs (2017) ."} +{"text": "Disrupted mitochondrial functions and genetic variants of mitochondrial DNA (mtDNA) have been observed in different human neoplasms. Next-generation sequencing (NGS) can be used to detect even low heteroplasmy-level mtDNA variants. We aimed to investigate the mitochondrial genome in pituitary adenomas by NGS.We analysed 11 growth hormone producing and 33 non-functioning [22 gonadotroph and 11 hormone immunonegative] pituitary adenomas using VariantPro\u2122 Mitochondrion Panel on Illumina MiSeq instrument. Revised Cambridge Reference Sequence (rCRS) of the mtDNA was used as reference. Heteroplasmy was determined using a 3% cutoff.496 variants were identified in pituitary adenomas with overall low level of heteroplasmy (7.22%). On average, 35 variants were detected per sample. Samples harbouring the highest number of variants had the highest Ki-67 indices independently of histological subtypes. We identified eight variants with different prevalences among different histological groups. T16189C was found in 40% of non-recurrent adenomas, while it was not present in the recurrent ones. T14798C and T4216C were confirmed by Sanger sequencing in all 44 samples. 100% concordance was found between NGS and Sanger method.NGS is a reliable method for investigating mitochondrial genome and heteroplasmy in pituitary adenomas. Out of the 496 detected variants, 414 have not been previously reported in pituitary adenoma. The high number of mtDNA variants may contribute to adenoma genesis, and some variants might associate with benign behaviour.The online version of this article (10.1007/s40618-019-1005-6) contains supplementary material, which is available to authorized users. The mitochondrial genome consists of several copies of circular, double-stranded DNA molecules, covering 16,569 base pairs, 37 genes. Of these, 13 encode polypeptides of respiratory enzyme complexes, 22 encode transfer RNAs, and 2 encode ribosomal RNAs mutations that frequently (approx. 40%) occur in somatotropinomas pituitary adenomas.The study was approved by the Scientific and Research Committee of the Medical Research Council of Hungary (0618/15), and the samples were obtained after acquiring written informed consent from all patients.Tissue samples were obtained from 44 patients diagnosed with pituitary adenoma, comprising 11 GH-secreting and 33 clinically non-functioning pituitary adenomas (NFPAs), including 22 gonadotroph (GO), and 11 hormone-immunonegative (HN) tumors . DNA purity and concentration were measured using NanoDrop 1000 Spectrophotometer (Thermo Fisher Scientific). The presence of normal pituitary tissue was determined similarly to V\u00e4lim\u00e4ki et al. . In our DNA library was prepared using the VariantPro\u2122 Mitochondrion Panel Library Preparation Kit (LC Sciences). The presence of the desired fragments and the purity of the indexed libraries were analysed on Agilent 2100 Bioanalyzer (Agilent Technologies) using High-Sensitivity DNA Analysis Kit (Agilent Technologies). The concentrations of the libraries were measured using Qubit Fluorometer (Thermo Fisher Scientific). Equimolar amounts of the 45 indexed libraries were pooled to obtain a 4\u00a0nM library mixture. After denaturing, and further diluting, the final 10\u00a0pM of library mixture was loaded into Illumina cartridge. Sequencing was performed using the Illumina MiSeq Reagent v2 kit (500 cycles) on the Illumina MiSeq instrument following the manufacturer\u2019s instructions (Illumina).http://www.bioinformatics.babraham.ac.uk/projects/trim_galore/) and cutadapt adenomas. HmtDB samples represent only GH positive adenomas (n\u2009=\u200919). Out of 496 variants 414 were firstly identified by the current study (DOCX 56\u00a0kb)Online Resource 2 Mitochondrial variants uniquely present in different adenoma types (DOCX 18\u00a0kb)Below is the link to the electronic supplementary material."} +{"text": "To test if mtDNA defects are a contributing factor in ESCC, we used oncogenic stimuli such as ESCC carcinogen 4-nitroquinoline oxide (4-NQO) treatment, or expressing p53R175H oncogenic driver mutation. We observed that EECs and 3D-organoids with mtDNA depletion had cellular, morphological and genetic alterations typical of an oncogenic transition. Furthermore, mitochondrial dysfunction induced cellular transformation is accompanied by elevated mitochondrial fission protein, DRP1 and pharmacologic inhibition of mitochondrial fission by mDivi-1 in the -/-MPV17 organoids reversed the phenotype to that of normal EEC organoids. Our studies show that mtDNA copy number depletion, activates a mitochondrial retrograde response, potentiates telomere defects, and increases the oncogenic susceptibility towards ESCC. Furthermore, mtDNA depletion driven cellular plasticity is mediated via altered mitochondrial fission-fusion dynamics.Esophageal squamous cell carcinoma (ESCC) is an aggressive cancer with late-stage detection and poor prognosis. This emphasizes the need to identify new markers for early diagnosis and treatment. Altered mitochondrial genome (mtDNA) content in primary tumors correlates with poor patient prognosis. Here we used three-dimensional (3D) organoids of esophageal epithelial cells (EECs) from the R175H or EGFR mutations [Esophageal Squamous Cell Carcinoma (ESCC) is one of the most aggressive squamous cell cancers and is the sixth leading cause of cancer-related mortality in the world . Esophagutations . Carcino2+ homeostasis and regulate apoptosis: cellular processes that are altered during oncogenic transformation. Functional interaction between mitochondria and nucleus controls both the biogenesis and functioning of mitochondria. Several cellular and environmental conditions disrupt mitochondrial function. Mitochondrial (mt) DNA mutations, deletions or impaired mtDNA replication are common causes of mitochondrial dysfunction. We have previously demonstrated that dysfunctional mitochondria alter the cytosolic Ca2+ pool and trigger a Ca2+-calcineurin dependent mitochondria-to-nucleus stress signaling pathway (MtRS) [Adaptation to the metabolic demands of proliferating cancer cells is critical to their survival, but is also their vulnerability. Mitochondria are cellular signaling hubs and integrate various metabolic pathways, synthesize intermediates required for the synthesis of biomass, maintain Cay (MtRS) \u201311. Actiy (MtRS) \u201310. It iy (MtRS) , 13. How-/-MPV17 which contains tissue-specific mtDNA depletion [Tfam-/+ mice [MPV17 is a mitochondrial inner membrane protein. Loss of MPV17 (-/-MPV17) causes mtDNA depletion and impairs oxidative phosphorylation (OXPHOS) in humans and mice, while the heterozygous (MPV17+/-) mice have mtDNA content and mitochondrial functions comparable to the wild type mice [Tfam is a mitochondrial transcription factor that controls mtDNA copy number. In most somatic tissues, Tfam levels correlate tightly with mtDNA content and Tfam heterozygous cells contain 50% reduced mtDNA while Tfam KO attain Rho0 state . These two models are therefore ideal to demonstrate the role of mtDNA defects and dysfunctional mitochondria in ESCCs. To study the ESCC oncogenic process ex vivo, we utilized the three-dimensional (3D) organoid system. 3D organoids provide a cell culture based, physiologically relevant, platform exhibiting tissue-like architecture grown in a mount of basement membrane extract with media containing the niche factors as described before [To determine the role of dysfunctional mitochondria in ESCC progression, we utilized two different mouse models of mtDNA depletion: 1) epletion , 15 and -/+ mice , 17. MPVype mice , 15. Tfad before . The mord before . The 3D d before , 20.-/-MPV17 model of mtDNA depletion we observed telomere defects and chromosomal defects typical of tumor cells. Further, in the -/-MPV17 organoids, we observed increased tumorigenic transformation and higher susceptibility to ESCC in response to oncogenic or carcinogenic stimuli. This is the first report that demonstrates the contribution of dysfunctional mitochondria towards 4NQO induced ESCC development using a novel murine mtDNA depletion 3D organoid model.We observed that primary cells and 3D organoids derived from esophageal epithelia of mice containing partial mitochondrial DNA (mtDNA) depletion shows the activation of the mitochondrial retrograde signaling (MtRS) pathway and cellular alterations that resemble oncogenic transition. In the MPV17+/+), MPV17 heterozygotes (+/\u2013) and homozygous knockout (\u2013/\u2013) mice . Single cells enzymatically dissociated from the mucosa were cultured ex vivo (referred henceforth as EEC) as described in Materials and Methods. In the -/-MPV17 mouse model, the mtDNA depletion is tissue specific [-/-MPV17 show 80% reduction in mtDNA content compared to WT analysis using telomeric DNA specific Cy-3 PNA probe showed marked loss of telomere signals (indicated by yellow arrows), higher telomeric signal-free ends at the chromatids and marked number of chromosome end-fusions in -/-MPV17 EECs which plays a causal role in telomere attrition similar to that observed in tumor cells . We therMPV17-/+ . Quantit-/- EECs . This suR175H [-/-MPV17 mice, human EPC2 cells [We further tested the contribution of mitochondrial stress in ESCC progression using human esophageal keratinocyte cell line EPC2-hTERT (EPC2) and EPC2-hTERT cells expressing the most prevalent ESCC mutation in gene TP53R175H . In agreC2 cells exhibit MPV17+/--/-or MPV17 mice were harvested and enzymatically dissociated into single cells and grown either as two-dimensional cultures, or were suspended in Matrigel\u2122 as detailed in Materials and Methods [The esophageal epithelial cells (EECs) from WT and Methods to gener-/-MPV17 EECs shows altered F-actin organization resulting in lamellipodia and filopodia membrane protrusion structures, and cells were markedly enlarged, typical of migratory cancer cells mice , which is our parallel model for mtDNA depletion showing the characteristic mtDNA content reduction and telomere attrition .Migration of cancer cells during metastatic transition is a complex and critical process requiring reorganization of actin filaments suggesting cytoskeletal remodeling. Prior studies have shown that actin and its interacting partners such as the Rho GTPases, along with downstream effector proteins mediate processes involved in tumor cell migration, invasion and metastasis through the cytoskeleton. Phalloidin staining of primary er cells . Phalloi-/- mice formed dysmorphic structures invading into the surrounding Matrigel\u2122 matrix typical of oncogenic phenotype for 8 weeks. This treatment duration is substantially shorter compared to the 16 weeks of 4NQO treatment followed by 12 weeks of observation used in other studies for inducing esophageal cancers. We observed that -/-MPV17 animals exhibited loss of body weight (nearly 20%) while there was no significant weight loss in either the WT or MPV17-/+ mice suggesting that 4NQO treatment had substantially higher deleterious effects on the -/-MPV17 mice . There were visible lesions in the esophagus of the -/-MPV17 mice while no abnormality was observed in esophagus in WT or MPV17-/+ mice. Histopathological analysis of the esophageal sections indicated pre-cancerous lesions including esophageal hyperplasia and dysplasia in -/-MPV17 mice without any abnormality in esophagi of WT mice. Mild hyperplasia with basal layer activity and some minor dysplasia indicated by a mild cell polarization alteration, some hyperchromatic nuclei and mild changes in nuclear size and shape were observed in -/-MPV17 mouse esophagi . Additionally, we observed basal/parabasal cell vacuolization in some areas consistent with observed effects of carcinogens .We observed that -/-MPV17 mice treated with 4NQO as described above. H&E staining of the organoid cross sections demonstrate that -/-MPV17 organoids have dysplastic mucosa consistent with neoplastic transformation (+/fl +adeno CRE (Tfam+/-) primary EECs were larger in size and had reduced mitochondrial membrane potential .ormation . Similar-/-MPV17 mice (-/-MPV17 mouse derived organoids showed higher expression of Ki-67 (intense staining) similar to esophageal tumors indicative of high proliferation as observed in neoplastic cells. The mitochondrial retrograde signaling (MtRS) pathway can be triggered by mtDNA depletion or by loss of nuclear encoded mitochondrial electron transport chain protein CcOIVi1, [-/-MPV17 organoids compared to WT organoids were non-invasive while WT cells expressing the oncogenic TP53R175H mutation acquired invasive capacity in telomere signals similar to our earlier observation in -/-MPV17+ TP53R175H EECs show a further 45% reduction compared to -/-MPV17 EECs and 75% loss in telomere signals compared to that in WT EECs . Our results suggest that mitochondrial stress provides an oncogenic stimulus, which augments the effects of oncogenic mutations to induce transition to cancer phenotype.We observed a similar pattern of invasiveness in the Tfam-/-MPV17 organoids. Therefore, we treated the organoids ex vivo with DRP1 inhibitor, mDivi-1 to inhibit mitochondrial fission and observed a reversal in the -/-MPV17 organoid morphology towards the normal phenotype suggestive of the involvement of higher fission in driving the cellular plasticity in this model of mitochondrial dysfunction and Dynamin-related-protein 1 (DRP1) . Our reser cells and that-/-MPV17 esophageal epithelium in less than 8 weeks. This shorter treatment duration required to induce carcinogenic alteration in -/-MPV17 compared to the WT or MPV17-/+ mice suggests that mitochondrial dysfunction is an additive factor in potentiating the carcinogenic effect of 4NQO. Furthermore, we observed that in human esophageal keratinocytes, chemically induced-mtDNA depletion resulted in loss of telomere length typical of tumor cells. This is in agreement with our recent report showing that mitochondrial stress plays a causal role in telomere attrition and chromosomal aberrations in skeletal myocytes [Epidemiological studies suggest an association with tobacco and alcohol abuse . Additiomyocytes . Other rmyocytes . A studymyocytes .Based on our data here, we propose that while mitochondrial dysfunction by itself may not be sufficient for ESCC initiation, the presence of carcinogenic and/or nuclear oncogenic stimulus increases ESCC susceptibility and can potentially dictate the tumorigenic progression and therapy outcome. We earlier reported that cigarette smoke toxins and environmental carcinogens have deleterious effects on mitochondrial functions , 46. IndThe MPV17 knock out mice used in this study were obtained from Jackson Laboratories and bred to BALB/c mice for 10 generations before using for experiments. At the initiation of this study, all animals used were age matched (8 weeks) and represented both sexes. Animals were housed and cared for in accordance with the regulations of the University of Pennsylvania\u2019s Institutional Animal Care and Use Committee. Mice were euthanized using CO2 asphyxiation using an IACUC approved protocol before harvesting tissues.The genotyping primers used are as follows:MPV1717 WT/FW AACCACTACGGCTGGCTAGA.MPV1717 WT/RC GCTTCAAAGCAAACGACCTC.MPV1717 MUT/RC CCTACAGGTGGGGTCTTTCA.mpv17 and tfam) as well as by genomic profiling.All tissue isolation from the MPV17 and Tfam mice were performed according the approved IACUC protocol (Protocol Number: 805731). Primary murine esophageal epithelial cells were harvested following standard protocol . BrieflyCell size and viability was measured by staining cells suspensions (20 \u03bcl) with 0.2% trypan blue and analyzed using an automated cell counter . Identical number of cells (between 1000\u20131500) were counted for each cell type.Esophageal keratinocytes were isolated from untreated, control treated or 4NQO- treated mice. Using 24-well plates, 5000 cells were seeded per well in 50 \u03bcl Matrigel. After solidification, 500 \u03bcl of DMEM/F12 supplemented with 1\u00d7 Glutamax, 1\u00d7 HEPES, 1\u00d7 N2 Supplement, 1\u00d7 B27 Supplement, 0.1 mM N-acetyl-cysteine (Sigma-Aldrich), 50 ng/ml mouse recombinant EGF (R&D Systems), 2.0% Noggin/R-Spondin-conditioned media and 10 \u03bcM Y-27632 were added and replenished every other day. Organoid formation rate was calculated as the percentage of the number of organoids formed at day 7 per total number of cells seeded at day 0. After 10 days, the organoids were recovered by digesting Matrigel with Dispase I and fixed overnight in 4.0% paraformaldehyde. Specimens were embedded in 2.0% Bacto Agar and 2.5% gelatin prior to paraffin embedding. Cross sections (50 \u00b5m) of the organoids were stained with hematoxylin-eosin. Immunohistochemistry using antibodies (as indicated) was performed on parallel sections.Analysis of MtDNA content from cellular total DNA was performed using real time qPCR as described before . Copy nuCOX1 gene:TGATCTGCTGCAGTGCTCTGA (forward).TCAGGCCACCTACGGTGAA (reverse).COX IV1i gene:GAAAGTGTTGTGAAGAGCGAAGAC (forward).GTGGTCACGCCGATCCAT (reverse).Total cellular RNA was prepared using the RNeasy mini kit\u2122 (Qiagen Cat # 74104). Genomic DNA was eliminated from the RNA preparations using Turbo DNA Free kit\u2122 (Thermo Fisher Scientific). 1 \u03bcg RNA was reverse transcribed into cDNA using High Capacity reverse transcription kit (Applied Biosystems). 100 ng cDNA was used for each SYBR Green reaction for transcript analysis of the genes as indicated. Quantitative Real Time PCR assays were run on an ABI Quant Studio 6 real time thermocycler (Applied Biosystems). The nuclear gene actin was used as an endogenous control. All real time PCR assays were run in triplicate. Data are presented as Relative Quantification (RQ).In vitro invasion assays were performed as described before [4) in growth medium were seeded on a Matrigel-coated Boyden chamber. After 24 h, cells that invaded the Matrigel were stained with hematoxylin-eosin and observed under bright field microscope.d before , 47. CelImmunostaining of human esophageal cancer tissue or mouse esophageal organoid sections was done using Vectastain ABC kit according to manufacturer\u2019s instructions. Briefly, sections were de-paraffinized and incubated in blocking buffer for 1 h at 37\u00b0C. Incubation with primary antibodies was carried out overnight at 4\u00b0C. Biotinylated secondary IgG incubation was then carried out at 37\u00b0C and the signal was developed using the DAP peroxidase staining kit (Vector laboratories).5 cells per well on a 6-well plate) were grown overnight in growth medium on Poly-D Lysine coverslips. Cell adherence and confluence was confirmed before processing. After 24 h of cell seeding, cells were washed with 1X PBS and fixed in ice-cold methanol for 10 minutes, at room temperature. Fixed cells were blocked in a buffer containing 1% BSA and were incubated with primary antibodies for 1 hour at 37\u00b0C. Immunostained cells were imaged using a Leica confocal microscope under a 100x objective.Cells for 3 hours and harvested for metaphase spreads. Cells were swollen in hypotonic solution at 37\u00b0C for 15 minutes, and fixed with methanol: acetic acid (3:1) with three repeated exchanges prior to dropping onto slides and dried overnight.Nuclei and metaphase spreads were processed for telomere Q-FISH. After washing and hypotonic swelling, cells were fixed and stored in methanol/acetic acid fixative using standard procedures . Nuclei The nuclei and metaphases on the PNA hybridized slides were visualized under a Nikon microscope and images were captured under 100x objective. The image acquisition conditions were kept identical for all cell types. For quantitation, the raw images of nuclei were used for analysis using MetaMorph software (Molecular Devices). Cy3-PNA signals were counted and the fluorescence intensity was quantitated by applying consistent intensity and size thresholds. The average DAPI fluorescence intensity for each nucleus was quantified and used to normalize the measured Cy3 PNA fluorescence intensities. The total DAPI fluorescence signal for each nucleus was quantified. At least 10 nuclei were counted for each cell type.Telomere-FISH: Cy3 PNA probe: Cy3-OO-CCCTAA CCCTAACCCTAA.t test. P value of < .05 was considered statistically significant. Statistical analyses were performed using Prism software .All experiments were performed in biologic and technical replicates. Data are presented as means \u00b1 SEM and statistical significance was determined using a 2-tailed unpaired Student"} +{"text": "Dear Editor,Autophagy is an intracellular degradation pathway regulated by the orchestrated action of the autophagy\u2010related (ATG) proteins. ATG proteins traditionally have been studied for their roles in autophagy, but they have increasingly demonstrated functions other than cellular self\u2010eating.+ (virus\u2010infected) cells than among cells transfected with empty vector with an IFN\u2010sensitive response element (ISRE) luciferase reporter, which was an interferon regulatory factor 3 (IRF3)\u2010dependent promoter, and the internal control renilla luciferase, as well as expression vectors containing ATG13. Then we treated the cells intracellularly with the synthetic nucleic acid duplex poly(I:C)/ poly(dA:dT) or infected with vesicular stomatitis virus (VSV) for 12\u00a0hours to trigger type I interferon signalling. The ISRE\u2010Luc activity induced by intracellular poly(I:C)/ poly(dA:dT) or VSV infection was greatly enhanced by HA\u2010ATG13 overexpression constructs to knock down the expression of ATG13. All three efficiently inhibited the expression of endogenous ATG13 in mRNA level and transfected ATG13 in protein level in 293T cells (Figure Altogether, our results suggest that ATG13 may regulate viral replication by potentiation type I interferon signalling. TARF3 and TRAF6 may be the important target of ATG13 to regulated type I interferon production. Further studies are needed to investigate the detailed mechanisms of ATG13 in the regulation of type I interferon.The authors confirm that there is no conflict of interest."} +{"text": "The baseline samples are of lagomorphs and rodents with limited foraging ranges. The baseline ranges for each site were calculated with two standard deviations. Also included are the raw strontium isotopic data for 30 large game samples from Wolf Village, a Fremont site in Utah. Additional data include a map showing the location of the sites in this study, box plots portraying the local ranges of nine Fremont sites in Utah, and an individual value plot comparing the Wolf Village large game samples to the strontium baseline for the site. These data compliment the discussions and interpretations found in \u201cIdentifying Strontium Baselines and Large Game Animal Trade at Fremont Sites through Strontium Isotope ( All areas had large Fremont settlements. Sites around Utah Lake include Hinckley Mounds (42UT111), Woodard Mound (42UT102), and Wolf Village (42UT273). In addition, Nephi Mounds (42JB02) is somewhat close to the Utah Lake sites and could be accessed by the Fremont through Goshen Canyon. Sites in central Utah include Nawthis Village (42SV633), Five Finger Ridge (42SV1686), and Icicle Bench (42SV1372). Sites in the Parowan Valley in southeastern Utah include Parowan (42IN100) and Paragonah (42IN43). The multiple sites in each of the three main areas can be compared to identify variability in local strontium levels.87Sr/86Sr ratios [To avoid contamination from modern fertilizers and air pollutions, rodents and lagomorphs from the archaeological record are useful to identify local r ratios . In this2.2Odocoileus hemionus) specimens, four pronghorn (Antilocapra americana) specimens, eight bighorn sheep (Ovis canadensis) specimens, and five worked bone gaming piece specimens constructed from long bones of unidentified large game (Artiodactyla) . All tes2.3All small mammal and large game samples were analyzed at the Strontium Isotope Geochemistry Laboratory at the University of Utah. Pretreatment was done at the Biogeochemistry Laboratory at the University of Utah. Small mammal samples are too small to manually remove dentine from enamel, so whole tooth specimens were analyzed. For large game samples, teeth were extracted from the mandible using a Dremel Lithium-Ion cordless drill . Samples were examined under a Bausch & Lomb StereoZoom 5 (zoom range 0.8\u00d7\u20134.0\u00d7) microscope to ensure the dentine and discoloration were removed from the samples, leaving as much tooth enamel as possible (at least 0.05 g). The Dremel Lithium-Ion drill was also used to remove at least 0.05 g from each of the worked bone gaming pieces.3COOH) and then rinsed three times in quadrupole de-ionized water (4\u00a0\u00d7\u00a0H2O). These methods are effective at removing contaminants from samples [3) in sterile Teflon vials. Strontium concentrations were determined by analyzing the digested samples in a quadrupole inductively coupled plasma mass spectrometer (ICP-MS) . To isolate strontium from other ions, a small portion of the digest (200 ng) was purified using column chromatography with resin Sr-Spec in an automated system . The purified Sr fraction was dried and then rehydrated with 1 mL of 5% HNO3. The samples were then analyzed on a Neptune Plus multi-collector ICP-MS . A certified reference material NIST SRM 987 was run every three samples. A blank was run after each SRM sample with the SRM value (87Sr/86Sr\u00a0=\u00a00.71028) being within the acceptable range of other analysts [Samples were pretreated with 5% acetic acid specimens may not reflect accurate strontium baselines since muskrats are semiaquatic animals. Muskrats may be influenced by nonlocal strontium coming from various geological formations through rivers and streams. Therefore, their bones and teeth may no longer represent local strontium baselines. Unfortunately, a previous baseline for Wolf Village was based on strontium levels in semiaquatic muskrat [Spermophilus sp.) incisors from Wolf Village were analyzed. The Wolf Village baseline using squirrels is more precise than the muskrat baseline and contains no outlier specimens outside the local strontium range (Muskrat ( muskrat . To testum range . Comparium range .Comparisons between the muskrat and squirrel baselines to the Wolf Village large game specimens provide differing results . Almost Charles Redd Center for Western Studies, the Grace Elizabeth Shallit Memorial Grant, the Warren Van Pelt Student Grant, and the Department of Anthropology at Brigham Young University. The Charles Redd Center for Western Studies funded the publication of this research.Funding for this project was generously provided by the"} +{"text": "Intradialytic hypotension is a serious complication during renal replacement therapy in critically ill patients. Early prediction of intradialytic hypotension could allow adequate prophylactic measures. In this study we evaluated the ability of peripheral perfusion index (PPI) and heart rate variability (HRV) to predict intradialytic hypotension.A prospective observational study included 36 critically ill patients with acute kidney injury during their first session of intermittent hemodialysis. In addition to basic vital signs, PPI was measured using Radical-7 (Masimo) device. Electrical cardiometry (ICON) device was used for measuring cardiac output, systemic vascular resistance, and HRV. All hemodynamic values were recorded at the following time points: 30\u2009min before the hemodialysis session, 15\u2009min before the start of hemodialysis session, every 5\u2009min during the session, and 15\u2009min after the conclusion of the session. The ability of all variables to predict intradialytic hypotension was assessed through area under receiver operating characteristic (AUROC) curve calculation.Twenty-three patients (64%) had intradialytic hypotension. Patients with pulmonary oedema showed higher risk for development of intradialytic hypotension {Odds ratio (95% CI): 13.75(1.4\u2013136)}. Each of baseline HRV, and baseline PPI showed good predictive properties for intradialytic hypotension {AUROC (95% CI): 0.761(0.59\u20130.88)}, and 0.721(0.547\u20130.857)} respectively.Each of low PPI, low HRV, and the presence of pulmonary oedema are good predictors of intradialytic hypotension. Acute kidney injury is common among critically ill patients. Intermittent hemodialysis is one of the commonly used routes for renal replacement therapy . IntradiPeripheral Perfusion Index (PPI) is defined as \u201cthe ratio of pulsatile blood flow to the non-pulsatile blood flow\u201d. PPI is measured using pulse co-oximetry technology which is characterized by being simple and non-invasive. PPI mirrors the strength of blood flow and quality of perfusion at sensor site, reflecting the global perfusion state of the body . As PPI Heart rate variability (HRV) is commonly described as a \u201cnew vital sign\u201d which had shown promise in evaluation of autonomic nervous system function . FurtherThe aim of this work was to evaluate the ability of PPI and electrical cardiometry-derived HRV to predict hypotension in critically ill patients during intermittent hemodialysis.A prospective observational trial was conducted in Cairo University hospital after Institutional Research Ethics approval N-91-2018) including a cohort of 36 adult critically ill patients with acute kidney injury (AKI). Written informed consent was obtained from patients or their surrogates before inclusion in the study. We included patients who were scheduled for first session intermittent hemodialysis according to Kidney Disease Improving Global Outcomes (KDIGO) guidelines ), pulmonary edema (6 patients [17%]), eclampsia (1 patient [3%]), diabetic ketoacidosis (2 patients [6%]), and disturbed conscious level due to head trauma (8 patients [22%]) or non-traumatic intracranial hemorrhage (4 patients [10%]). Four (11%) patients were mechanically ventilated and four (11%) patients were on norepinephrine infusion before starting the hemodialysis session. Twenty-three (64%) patients had intradialytic hypotension. Demographic data and patient chronic comorbidities were comparable between hypotensive group and stable group; whilst, APACHE II score was higher in the hypotensive group compared to stable group : 0.761(0.59\u20130.88), cut-off value \u226424}, and {AUROC (95% CI): 0.721(0.547\u20130.857), cut-off value \u22641.8} respectively. PPI was superior in terms of sensitivity and negative predictive value (NPV) (100 and 100%); whilst, HRV was superior in terms of specificity and positive predictive value (PPV) (91 and 92%). and intradialytic hypotension with Odds ratio of 13.75(1.4\u2013136). Volume overload had been considered as a biomarker for the severity of critical illness . Fluid oOur study has the advantage of using simple non-invasive variables. Furthermore, we evaluated patients during the usual route for renal replacement therapy which is intermittent hemodialysis. Intradialytic hypotension is a common and serious complication during hemodialysis. Early detection of high-risk patients for intradialytic hypotension would help to decrease their risk through early initiation of vasopressors. High filtration rate and ultrafiltration volume are important factors which contribute in intradialytic hypotension ; Hence, Low PPI, low HRV, and the presence of pulmonary oedema are useful predictors of intradialytic hypotension."} +{"text": "Cyprinodon pupfish species\u2013a dietary generalist and a specialized molluscivore\u2013and measured expression levels in their F1 hybrids to identify regulatory variation underlying the novel craniofacial morphology found in this recent microendemic adaptive radiation. We extracted mRNA from eight day old whole-larvae tissue and from craniofacial tissues dissected from 17\u201320 day old larvae to compare gene expression between a total of seven F1 hybrids and 24 individuals from parental species populations. We found 3.9% of genes differentially expressed between generalists and molluscivores in whole-larvae tissues and 0.6% of genes differentially expressed in craniofacial tissue. We found that 2.1% of genes were misregulated in whole-larvae hybrids whereas 19.1% of genes were misregulated in hybrid craniofacial tissues, after correcting for sequencing biases. We also measured allele specific expression across 15,429 heterozygous sites to identify putative compensatory regulatory mechanisms underlying differential expression between generalists and molluscivores. Together, our results highlight the importance of considering misregulation as an early indicator of genetic incompatibilities in the context of rapidly diverging adaptive radiations and suggests that compensatory regulatory divergence drives hybrid gene misregulation in developing tissues that give rise to novel craniofacial traits.Genetic incompatibilities constitute the final stages of reproductive isolation and speciation, but little is known about incompatibilities that occur within recent adaptive radiations among closely related diverging populations. Crossing divergent species to form hybrids can break up coadapted variation, resulting in genetic incompatibilities within developmental networks shaping divergent adaptive traits. We crossed two closely related sympatric Changes in gene expression are an important source of variation in adaptive morphological traits \u20133. As geOf particular importance to the process of speciation are genetic incompatibilities caused by hybrid misregulation\u2013transgressive expression levels in hybrids that are higher or lower than both parental species ,11\u201315. Tcis-regulatory elements and trans-regulatory factors. Cis elements are often non-coding regions of DNA proximal to genes that are bound by trans-acting proteins and RNAs to regulate mRNA abundance. It is possible to identify mechanisms of gene expression divergence between parental species by bringing cis elements from both parents together in the same trans environment in F1 hybrids and quantifying allele specific expression (ASE) of parental alleles at heterozygous sites [Cis and trans regulatory variants can compensate for one another if stabilizing selection favors an optimal level of gene expression. Hybrid misregulation is expected when different compensatory variants have accumulated in diverging lineages [Studies of gene expression in hybrids can also implicate regulatory mechanisms underlying expression divergence between parental species, which is important for understanding how expression levels are inherited and how they shape adaptive traits \u201324. Reseus sites ,29. Cis lineages \u201333.Cyprinodon pupfishes to understand regulatory mechanisms that led to the evolution of novel craniofacial adaptations in this group following approved protocols from the University of California, Davis Institutional Animal Care and Use Committee (#17455) and University of California, Berkeley Animal Care and Use Committee (AUP-2015-01-7053). Field collections were permitted by the Bahamian Department of Agriculture (permit agr/nat/1).Our methods for raising larvae and extracting RNA were identical to previously outlined methods . We collWe previously generated 24 transcriptomes belonging to generalists and molluscivores collected at two early developmental stages: 8\u201310 days post fertilization (dpf) and 17\u201320 dpf . RNA wasHere we analyze an additional 19 transcriptomes from generalists, molluscivores, and their F1 hybrids . First, We performed separate crosses to collect larvae at exactly 8 dpf (190\u2013194 hours after fertilization rather than 8\u201310 days). We crossed a generalist female with a molluscivore male to generate three F1 hybrids for whole-larvae RNA extractions. The parents of these hybrids were wild-caught from Osprey Lake. Finally, we extracted whole-larvae RNA from six generalists and six molluscivores collected at 8 dpf. These samples were generated from wild-caught individuals from Osprey Lake and Crescent Pond. In total, we analyzed transcriptomes from 43 individuals that involved four separate rounds of sequencing .The previously reported 24 transcriptomes were sequenced at the High Throughput Genomic Sequencing Facility at UNC Chapel Hill in April 2017 . The 24 19 additional transcriptomes were sequenced at The Vincent J. Coates Genomics Sequencing Laboratory at the University of California, Berkeley. All 19 libraries were prepared at the facility using the Illumina stranded Truseq RNA kit (Illumina RS-122-2001) and all sequencing was performed on Illumina 150 paired-end Hiseq4000. Four libraries for RNA extracted from 17\u201320 dpf hybrid craniofacial tissues were pooled on a single lane and sequenced in June 2017. 15 libraries for whole-larvae RNA samples collected at exactly 8 dpf were pooled across one and three lanes and sequenced in May (n = 9) and July (n = 6) 2018, respectively .Cyprinodon reference genome , but we did find that fewer reads mapped to features in 17\u201320 dpf samples than 8 dpf samples .We filtered all raw reads using Trim Galore to remove Illumina adaptors and low\u2010quality reads (mean Phred score < 20) and mapped filtered reads to the scaffolds of the 20,803; ). We use 20,803; requirin 20,803; . We preve genome . Similarin vitro RNA degradation within a sample. TIN is calculated by analyzing the uniformity of coverage across transcripts [Since we analyzed RNA from 43 individuals that were sequenced across four different dates and their libraries were prepared with either KAPA or TruSeq stranded mRNAseq kits, we tested whether a significant amount of between-sample variance in read counts was explained by sequencing date or library preparation kit. We fit linear models (using the lm function in R) to determine whether normalized counts across individuals were influenced by 1) the number of duplicate reads, 2) the uniformity of coverage across a transcript, or 3) the depth of coverage across GC-rich transcripts. All of these measures could have been influenced by different library preparation methods \u201356. RseQnscripts ,59. We pWe performed differential expression analyses with DESeq2 (v. 3.5 ). This pWe conducted pairwise comparisons to identify genes differentially expressed between hybrids vs. parental species, hybrids vs. generalists, hybrids vs. molluscivores, and generalists vs. molluscivores. \u201cParental species\u201d refers to generalists and molluscivores derived from the same populations as the parents of the hybrid samples. We did not sequence any of the parents crossed to generate hybrids. We defined genes as misregulated in hybrids if they were significantly differentially expressed between hybrids and the parental species samples. First, we compared whole-larvae gene expression between samples collected at 8 dpf . All of the 8 dpf samples were sequenced at the Vincent J. Coates Genomic Sequencing Laboratory, University of California Berkeley (VJCGSL UCB) and their libraries were all prepared using the TruSeq stranded mRNAseq kit. Second, we compared craniofacial tissue gene expression between samples collected at 17\u201320 dpf . The generalist and molluscivore samples were sequenced at the High-Throughout Sequencing Facility, University of North Carolina Chapel Hill (HTSF UNC) and their libraries were prepared using the KAPA stranded mRNA-seq kit, while the hybrids were sequenced at the VJCGSL UCB and their libraries were prepared using the TruSeq kit. In order to understand how sequencing at different facilities and using different library prep methods affected the proportion of genes misregulated between hybrids and parental species at 17\u201320 dpf, we performed a third set of comparisons between hybrids collected at 8 dpf (sequenced at VJCGSL UCB with TruSeq) and generalists and molluscivores from a previous study collected at 8\u201310 dpf . We meaTo determine whether genes showed additive, dominant, or transgressive patterns of inheritance, we quantified differences in gene expression between hybrids vs. parental species and compared them to genes differentially expressed between generalists vs. molluscivores . Hybrid Cyprinodon reference genome is annotated for genomic features enrichment analyses for genes differentially expressed between species and misregulated in hybrids that shared the same name as zebrafish orthologs using GO Consortium resources available at geneontology.org [The ase 100, ), and malogy.org . We searP-value using Fisher's exact test to detect strand bias > 60, Mann\u2013Whitney rank-sum test for mapping qualities (z > 12.5), Mann\u2013Whitney rank-sum test for distance from the end of a read for those with the alternate allele (z > 8.0). We used the VariantsToTable function (with genotypeFilterExpression \"isHet = = 1\") to output heterozygous variants for each individual. We counted the number of reads covering heterozygous sites using the ASEReadCounter . In total we identified 15,429 heterozygous sites across all 32 individuals with sequencing coverage \u2265 20\u00d7 that fell within 3,974 genes used for differential expression analyses. At the 8 dpf stage, we found 2,909 of the 3,974 genes that contained heterozygous sites common to all samples. At the 17\u201320 dpf stage, we found 2,403 genes containing heterozygous sites common to all samples.We followed the best practices guide recommended by the Genome Analysis Toolkit (v. 3.5 ) in ordeP < 0.05; [We assigned each heterozygous allele as the reference allele, alternate allele, or second alternate allele and matched each allele to its corresponding read depth. This allowed us to identify allele specific expression (ASE) by measuring expression variation between the two sites. We only measured ASE at sites that were heterozygous in all samples in each stage in order to account for differences in nucleotide diversity within populations . We used < 0.05; ,24). We < 0.05; , which uA common approach to identify regulatory mechanisms underlying expression divergence is to measure ASE at sites that are heterozygous in hybrids and alternately homozygous in parental species ,25. HoweWe previously found 1,014 genes differentially expressed in whole-larvae tissue between six generalists and six molluscivores collected 8\u201310 days post fertilization . Here wWe previously found 120 genes differentially expressed in craniofacial tissue between species at 17\u201320 dpf . Here weP = 8.5 \u00d7 10\u22125). Of the 370 genes showing misregulation, 138 were annotated as zebrafish orthologs used for gene ontology enrichment analyses. The only significantly overrepresented term was cellular lipid metabolic process (GO:0044255).We compared gene expression in whole-larvae tissue collected at 8 dpf from generalist and molluscivore populations (n = 12) with expression in their F1 hybrids (n = 3) and found that 370 out of 17,705 genes (2.1%) were misregulated in hybrids . SlightlThe majority of genes showed conserved levels of expression with no significant difference between hybrids and parental species (84.9%). In line with other hybrid expression studies ,27,28, mP < 2.2 \u00d7 10\u221216). Of the 6,590 genes showing misregulation, 2,876 were annotated as zebrafish orthologs used for gene ontology enrichment analyses. Misregulated genes were enriched for 210 ontologies, including embryonic cranial skeleton morphogenesis with expression in their F1 hybrids (n = 4) and found extensive hybrid misregulation. More than half of genes ) were differentially expressed in hybrids compared to parental species expression . There wAll of the 8 dpf samples were sequenced at the same facility using the same library preparation kit. However, the 17\u201320 dpf generalist and molluscivore samples were sequenced at a different facility than the 17\u201320 dpf hybrid samples and used a different library preparation kit. We took two approaches toward understanding how sequencing at different facilities and using different library kits may have affected the proportion of genes misregulated between hybrids and parental species at 17\u201320 dpf.First, we performed another differential expression comparison between whole-larvae hybrids collected at 8 dpf and whole-larvae parental species that we collected for a previous study between 8\u201310 dpf . The 8 dP > 0.05). When we grouped samples by sequencing date rather than library preparation method, we found that the 17\u201320 dpf hybrid craniofacial samples (sequenced 6/17) did not show any difference in median GC content, raw read counts, or raw fastq reads compared to samples sequenced on different dates (P < 0.01). TINs quantify the uniformity of coverage across transcripts and are informative as a measure of in vitro RNA degradation, which likely suggests that hybrid craniofacial samples experienced more degradation than other samples prior to sequencing. Indeed, lower TIN was significantly correlated with a lower number of normalized counts across samples (P = 2.0 \u00d7 10\u22125). We found approximately the same number of genes overexpressed in hybrids (25.83%) as there were genes underexpressed (25.77%), suggesting that many genes were overexpressed in hybrids despite potential RNA degradation.We also investigated whether a significant amount of between-sample variance in read counts was explained by library preparation method or sequencing date. For each sample we quantified the number of normalized read counts, raw read counts, and raw fastq reads. We also estimated the proportion of duplicate reads out of total mapped reads, the median percent GC content across mapped reads, and the uniformity of coverage across mapped reads (median transcript integrity numbers (TINs)). All of these measures could be influenced by different library preparation methods \u201356. Howent dates . Howeverin vitro RNA degradation than other samples, but this did not produce a bias toward more genes showing underdominant expression in hybrids was biased due to differences in the number of duplicate reads produced by two different library preparation methods . We quan hybrids .If a gene shows similar gene expression levels between parental species but shows biased allelic expression only in hybrids, it may be regulated by compensatory variation, and such genes are likely to be misregulated in F1 hybrids ,32. We iWe measured ASE across sites within 2,770 genes that showed no difference in expression between generalists and molluscivores at 8 dpf. We found 157 genes (5.4%) that were likely regulated by compensatory mechanisms, which showed ASE only in hybrids and were not differentially expressed between generalists and molluscivores. Of these, nine genes (0.33%) also showed misregulation in hybrids . We alsoWe also found more genes showing compensatory regulation in 17 dpf tissues than 8 dpf tissues using a more conservative method to identify genes showing ASE with MBASED . At 8 dpP = 2.81 \u00d7 10\u22125). Since misregulation is expected in hybrids when gene expression is controlled by compensatory variation between parental species [We found many more genes showing ASE in 17\u201320 dpf hybrid craniofacial tissue than any other samples , which could increase variance in the abundance of reads at heterozygous sites and bias ASE estimates. Lower TIN was correlated with higher ASE (P = 9.04 \u00d7 10\u221214). This correlation persisted when 17\u201320 dpf hybrid craniofacial samples were excluded from the model (P = 0.034), suggesting that rates of mRNA degradation may differ depending on genotypes at heterozygous sites. While this explains some of the elevated ASE in 17\u201320 dpf hybrid craniofacial samples, the proportion of genes showing ASE was much higher in these samples than predicted by the latter linear model. Even the lowest TIN for a 17\u201320 dpf hybrid sample (32.68) predicted a much lower range of genes showing ASE (8.2% -14.1%) compared to the observed range (32.8% - 51.6%). Finally, we also estimated ASE again with a higher coverage threshold to reduce the chances of increased variance affecting binomial tests and still found that hybrid craniofacial samples showed more ASE than other samples (P = 3.85 \u00d7 10\u22124).We tested whether this pattern might be due to higher rates of st = 0.08, Dxy = 0.00166; [Molluscivores show extreme craniofacial divergence relative to their generalist sister species, exhibiting a novel maxillary protrusion and short robust jaws . DesWhile many studies on hybrid misregulation search for regulatory divergence in \u2018speciation genes\u2019 associated with sterility and inviability ,14,19,24in vitro RNA degradation than other samples [There are several reasons why we might expect to find a higher proportion of genes misregulated in 17\u201320 dpf hybrid craniofacial tissues relative to 8 dpf whole-larvae tissues. The molluscivore shows exceptional rates of morphological diversification, particularly in craniofacial traits . Perhaps samples . While iDrosophila species pair [We found roughly twice the amount of bias-corrected misregulation in hybrid craniofacial tissues compared to a study of misregulation in whole-larvae tissue that measured gene expression in F1 hybrids generated between benthic and limnetic lake whitefish ,67. Thesies pair .It is unclear whether such extensive gene misregulation in hybrid craniofacial tissues might contribute to intrinsic postzygotic isolation between generalists and molluscivores. F2 hybrids exhibiting intermediate and transgressive craniofacial phenotypes showed reduced survival and growth rates in the wild relative to F2 hybrids resembling parental species ,46, but When an optimal level of gene expression is favored by stabilizing selection, compensatory variation can accumulate between species and cause misregulation in hybrids ,33. We cWe found hybrid misregulation in both whole-larvae tissues and craniofacial tissues sampled at early developmental stages. This points to divergent evolution of developmental networks shaping novel traits in the molluscivore. It is unclear whether such misregulation causes intrinsic incompatibilities in hybrids within this recent adaptive radiation. Our results are in line with studies finding widespread compensatory evolution in other systems with greater divergence times between species ,31,70,71S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.S4 TableP < 0.05; geneontology.org).Skeletal system morphogenesis (GO:0048705) was the only enriched biological process for genes differentially expressed between generalists and molluscivores at 8 dpf ((DOCX)Click here for additional data file.S5 TableP < 0.05; geneontology.org).Embryonic cranial skeleton morphogenesis (GO:0048701) was one of 210 enriched biological processes for 6,590 genes differentially expressed between hybrids and parental species in craniofacial tissue collected at 17\u201320 dpf ((DOCX)Click here for additional data file.S6 Table(DOCX)Click here for additional data file.S1 Fig20 day old generalist (top) and molluscivore (bottom).(PDF)Click here for additional data file.S2 FigProportion of reads assigned to features (yellow), unassigned due to multi-mapping (red), and unassigned due to no match to annotated features (blue) using STAR aligner.(PDF)Click here for additional data file.S3 FigThe first and second principal component axes accounting for a combined 75% of the total variation between generalist (red), molluscivore (blue), and hybrid (purple) samples across reads mapped to annotated features. Point shape indicates the sequencing date of the sample.(PDF)Click here for additional data file.S4 FigP = 8.5 \u00d7 10\u22125 17\u201320 dpf P < 2.2 \u00d7 10\u221216).Genes showing underdominant expression in hybrids show a higher magnitude of misregulation than genes showing overdominance Click here for additional data file.S5 FigThe 8 dpf hybrids were sequenced at the same facility with the same library kit as the 17\u201320 dpf hybrids, while the 8\u201310 dpf parental species were sequenced at the same facility with the same library kit as the 17\u201320 dpf parental species. (A) The comparison between 8 dpf parental species and 8 dpf hybrids revealed 370 genes (2.1%) misregulated. (B) The comparison between 8 dpf hybrids and 8\u201310 dpf parental species revealed 997 (6%) genes misregulated\u2013a 37% increase. We used this inflated estimate to adjust our estimate of misregulation in 17\u201320 dpf hybrid craniofacial tissues. Red points indicate genes detected as differentially expressed at 5% false discovery rate with Benjamini-Hochberg multiple testing adjustment. Grey points indicate genes showing no significant difference in expression between groups.(PDF)Click here for additional data file.S6 FigP < 0.0001 = ****, *** = 0.001, ** = 0.01, * = 0.05).We did not find significant differences between 17\u201320 dpf hybrid craniofacial samples and samples sequenced on other dates for (A) median percent GC content across reads, (B) number of normalized read counts, or (C) number of raw fastq reads. the proportion of duplicate reads for each sample Click here for additional data file.S1 AppendixExamples of command line arguments and DESeq2 scripts for bioinformatics pipelines.(DOCX)Click here for additional data file."} +{"text": "Prunus mira, we performed an integrated analysis of the transcriptome and metabolome of 3 fruit types with various flesh pigmentations at 3 developmental stages .Flesh color is one of the most important traits for the commercial value of peach fruit. To unravel the underlying regulatory network in core transcriptome\u2019 associated with major differentiations between the 3 fruit types, including flesh pigmentation. Meanwhile, we analyzed the metabolome, particularly, at the ripening stage and uncovered 40 differential metabolites (\u2018core metabolome\u2019) between the 3 fruit types including 5 anthocyanins, which may be the key molecules associated with flesh coloration. Finally, we constructed the regulatory network depicting the interactions between anthocyanins and important transcripts involved in fruit flesh coloration.Transcriptome analysis showed that an intense transcriptional adjustment is required for the transition from the pit-hardening to the cell enlargement stage. In contrast, few genes were differentially expressed (DEGs) from the cell enlargement to the fruit ripening stage and importantly, the 3 fruits displayed diverse transcriptional activities, indicating that difference in fruit flesh pigmentations mainly occurred during the ripening stage. We further investigated the DEGs between pairs of fruit types during the ripening stage and identified 563 DEGs representing the \u2018P. mira were unraveled in this study providing valuable information which will undoubtedly assist in breeding towards improved fruit quality in peach.The major metabolites and transcripts involved in fruit flesh coloration in Prunus. In China, P. mira has special nutritional, economic, medicinal and ornamental values http://ww.fna.gz) database.fna.gz) was used.fna.gz) .www.metware.cn) following their standard procedures and previously described by Wang et al. [The sample preparation, extract analysis, metabolite identification and quantification were performed at Wuhan MetWare Biotechnology Co., Ltd. , was used to maximize the metabolome difference between the developmental stages, as well as the difference between the three flesh colored fruits. The relative importance of each metabolite to the PLS-DA model was evaluated using the variable importance in projection (VIP). Metabolites with VIP\u2009\u2265\u20091 and fold change \u22652 or fold change \u22640.5 were defined as differentially accumulated metabolites between compared samples [First, a quality control (QC) analysis was performed to verify the reliability of the data. Sample extracts were mixed and inserted into every 10 samples to monitor the changes in repeated analyses. Metabolomics data have the characteristics of high dimension dataset, so it is necessary to combine univariate and multivariate statistical analyses to accurately excavate the differential metabolites. Statistical analyses were performed using the Analyst 1.6.1 software . Statistical significance and fold change of the metabolites between the samples were tested with two-paired samples , 58.2 transformed datasets were loaded in the \u2018cor\u2019 package from the R software (www.r-project.org/). The Pearson correlation (r) between metabolites and transcripts was represented by network diagrams, and the differential genes and metabolites between PMHF, PMHR and PMHY were selected when R2\u2009>\u20090.9 [For the joint analysis between the metabolome and transcriptome datasets, the mean of all biological replicates of differential metabolites in the metabolome data and the mean value of expression of differential transcripts in the transcriptome data were calculated. Next, the logR2\u2009>\u20090.9 . MetabolR2\u2009>\u20090.9 .RNAs from flesh samples were extracted using the EASYspin Plus kit according to the manufacturer\u2019s instructions. The RNA was treated with DNaseI and 1\u2009\u03bcg RNA was reverse transcribed with oligo (dT23) primer using the FastQuant RT kit in a final volume of 25\u2009\u03bcL. The specific primer pairs of the ten selected genes were designed using the Primer5.0 software and presAdditional file 1: Table S1. Overview of the transcriptome sequencing dataset and quality check in 3 Prunus mira fruit types during 3 developmental stages (pit-hardening (A), cell enlargement (B) and fruit ripening (C)). (XLSX 10 kb)Additional file 2: Table S2. Full list of the unique genes detected in Prunus mira fruit transcriptome and their functional annotation. (XLSX 2092 kb)Additional file 3: Table S3. List of the novel genes specific to Prunus mira detected from fruit transcriptome and their functional annotation. (XLSX 109 kb)Additional file 4: Table S4. List of the optimized genes based on gene structure optimization analysis in P. persica. (XLSX 301 kb)Additional file 5: Table S5. Intron-exon structure of the optimized genes in P. persica. (XLSX 3036 kb)Additional file 6: Table S6. List of all the differentially expressed genes identified in this study along with their FPKM data. (XLSX 4901 kb)Additional file 7: Figure S1. KEGG enrichment analysis of the 343 genes constantly and differentially expressed during the fruit development and ripening in Prunus mira. (TIF 522 kb)Additional file 8: Figure S2. KEGG enrichment analysis of the 3736 genes constantly and differentially expressed during the transition from the pit-hardening to the cell enlargement stages in Prunus mira. (TIF 565 kb)Additional file 9: Figure S3. KEGG enrichment analysis of the 607 genes constantly and differentially expressed during the transition from the cell enlargement to the fruit ripening stages in Prunus mira. (TIF 556 kb)Additional file 10: Figure S4. qRT-PCR (2-\u0394\u0394ct) analysis of 10 selected genes within the differentially expressed genes detected in this study. Correlation analysis between qRT-PCR and RNA-seq (log2fold change). (TIF 886 kb)Additional file 11: Table S7. List of the differentially expressed genes between the 3 Prunus mira fruit types representing the \u2018core transcriptome\u2019 and their functional annotation. (XLSX 26 kb)Additional file 12: Figure S5. KEGG enrichment analysis of the 563 genes differentially expressed in PMHF, PMHR and PMHY during the fruit ripening stage, representing the \u2018core transcriptome\u2019 in Prunus mira.s (TIF 522 kb)Additional file 13: Table S8. List of the differentially expressed genes between the 3 Prunus mira fruit types mapped to the terpernoid-carotenoid pathways and phenylpropanoid-flavonoid pathways and their expression level during the fruit ripening stage (C). (XLSX 11 kb)Additional file 14: Table S9. List of the bHLH and MYB transcription factors detected in the \u2018core transcriptome\u2019 and their expression fold change in group comparisons between the 3 Prunus mira fruit types during the fruit ripening stage. (XLSX 10 kb)Additional file 15: Table S10. Overview of the metabolites detected and quantified in 3 fruit types of Prunus mira during 3 developmental stages (pit-hardening (A), cell enlargement (B) and fruit ripening (C)). Data are from 3 biological replicates and the mix01 to mix05 represent the mixture of sample extracts. (XLSX 138 kb)Additional file 16: Table S11. List of the differential accumulated metabolites between the 3 Prunus mira fruit types representing the \u2018core metabolome\u2019 and their expression fold change between group comparisons during the fruit ripening stage. (XLSX 15 kb)Additional file 17: Table S12. List of genes showing a high correlation (R2\u2009>\u20090.9) with the 5 anthocyanins detected as important molecules involved in the fruit flesh coloration in Prunus mira. (XLSX 68 kb)Additional file 18: Table S13. The primer sequences for real time PCR. (XLSX 9 kb)"} +{"text": "Mashua virus Y (MasVY), first isolated in 1984 from a plant of the Andean tuber crop mashua . There was a 70% nucleotide identity between MasVY and a genomic sequence of Verbena virus Y.We present the complete genomic sequence of a new potyvirus we tentatively call Mashua virus Y (MasVY), first isolated in 1984 from a plant of the Andean tuber crop mashua . There was a 70% nucleotide identity between MasVY and a genomic sequence of Verbena virus Y.We present the complete genomic sequence of a new potyvirus we tentatively call Tropaeolum tuberosum, family Tropaeolaceae) following the manufacturer\u2019s instructions. The indexed library was sequenced on a MiSeq instrument (Illumina) with a 600-cycle v3 kit. The resulting 748,695 paired reads were 3\u2032 trimmed to a quality score of 20 with Sickle in paired-end mode (Verbena virus Y (VVY) (GenBank accession number NC_010735), from the ornamental plant Verbena \u00d7 hybrid (family Verbenaceae) reported from the United States (Potyviridae (Mashua virus Y (MasVY), and the three mashua potyviruses named previously including optional DNase treatment, total RNA was extracted from infected leaf material from yviridae . Establiviously 35 awaits MH680824 (genome) and MH680823 . Raw data were deposited in the SRA under BioSample number SAMN10081141, which is part of BioProject PRJNA491634.The sequences described here were deposited in GenBank under accession numbers"} +{"text": "Pharmacological treatment of recombinant growth differentiation factor 15 (GDF15) proteins reduces body weight in obese rodents and primates. Paradoxically, circulating GDF15 levels are increased in obesity. To investigate the role of endogenous GDF15 in obesity development, we put GDF15 knockout mice and wildtype controls on high fat diet for the mice to develop diet-induced obesity. Compared to wildtype animals, GDF15 knockout mice were more prone to high fat diet-induced obesity. Male knockout mice showed worse glucose tolerance, lower locomotor activity and lower metabolic rate than wildtype mice. Additionally, GDF15 deficiency increased occurrences of high fat diet-induced skin lesions. Our data suggests that endogenous GDF15 has a protective role in obesity development and lack of GDF15 aggravates the progression of obesity and associated pathological conditions. Elevated GDF15 levels in obesity may have resulted from a response to overcome GDF15 resistance. Obesity is a major public health burden associated with life-threatening comorbidities , 2. The The biological function of endogenous GDF15 is not completely clear. GDF15 knockout mice have been reported to be slightly heavier than wildtype mice , and exhEpidemiology and laboratory studies have demonstrated that increased fat intake is an important contributing factor to the obesity pandemic , 23, 24.All rodent studies were approved by Amgen Institutional Animal Care and Use Committee (IACUC) and conducted at Amgen Inc. . Animals were maintained in rooms with a 12-h light/dark cycle, temperature 22\u00baC and humidity 30% to 70%. Animals had free access to food and water and were maintained on standard rodent chow ) unless otherwise indicated.For high fat diet feeding studies, animals were maintained on 60% kcal fat diet .5\u2019-TCCCACATCAGCTGTCAGTC-3\u2019, wildtype reverse primer 5\u2019-CTACACCCCGGTGGTTCTTA-3\u2019, neo cassette reverse primer 5\u2019-CGTTGGCTACCCGTGATATT-3\u2019. PCR product of wildtype allele is 418bp. PCR product of knockout allele is 540bp.GDF15 constitutive knockout mice on a mixed 129S x C57Bl/6 background were acquired from Taconic Knockout Repository . Animals were bred to C57BL/6NTac background to be fully congenic (Charles River). GDF15 gene has 2 exons. A neo cassette was inserted into exon 2 to generate the knockout mice. Genotyping of GDF15 knockout mice was conducted by PCR analysis of DNA from ear samples using 3 PCR primers: common forward primer Circulating GDF15 levels were measured using Mouse/Rat GDF-15 Quantikine ELISA Kit (R&D systems)Body weight was measured every 2 weeks in home cages. For food intake measurements, clean cages with pre-weighed food were prepared, animals were transferred to these cages at regular cage-changing time, food intake was measured twice per week for 2 weeks.Fat mass, lean mass and fluid in conscious mice were measured using TD-NMR minispec body composition analyzer (Bruker)Blood glucose levels were measured using AlphaTrak 2 glucose strip (Abbott Diagnostics). Serum insulin levels were measured using mouse insulin ELISA kit (Alpco)Animals were fasted for 4 hours before baseline blood glucose measurement, blood sampling and an oral glucose bolus load . After glucose challenge, blood glucose levels were measured at 15, 30, 60 min.Indirect calorimetry was conducted using a 12-chamber comprehensive lab animal monitoring (CLAMS) Oyxmax system (Columbus). Animals were acclimated to the chambers for up to 2 weeks for body weight to stabilize prior to data collection. During the study, samples were taken at 13-min interval for measurement of O2 consumption and CO2 production, and calculation of respiratory exchange rate (RER) and heat production. Food intake was recorded every 13 min as changes of weight of the feeder. Locomotor activity was measured by breakings of infrared beams.Statistical analysis was performed using Graph Pad Prism software , and P<0.05 was considered statistically significant.At 9 weeks of age, the wildtype and knockout mice had similar body weights. We divided the animals into 2 sets. One set of animals continued with normal chow and the other set of animals were put on high fat diet. Body weight was measured every 2 weeks. Only 4 weeks after high fat diet feeding, we could start to observe significant differences between the male GDF15 knockout mice and wildtype mice . Male GDTo test if GDF15 deficiency affects glucose metabolism, we measured 4hr fasting blood glucose levels, serum insulin levels and conducted an oral glucose tolerance test on the knockout and wildtype mice after 9 weeks of high fat diet feeding. Male GDF15 knockout DIO mice had higher glucose levels and highAs we continued the longitudinal body weight measurements, we observed that female knockout DIO mice also became heavier than wildtype DIO mice, although the differences between genotypes in the female mice were not as robust as the differences observed in the male mice . We alsoTo further investigate if GDF15 deficiency promotes obesity development through regulation of energy substrate utilization or energy expenditure, we put wildtype and GDF15 knockout DIO mice in the Oxymax-CLAMS system and monitored food intake, locomotor activity, O2 consumption and CO2 production for 3 days after acclimation.Male GDF15 knockout DIO mice showed significantly lower locomotor activities during the dark cycle than the wildtype animals, measured as breakings of the X axis infrared beams for horizontal ambulatory movements and as bWe next analyzed metabolic rate and RER using O2 consumption and CO2 production measured in the Oxymax chambers. Male GDF15 knockout DIO mice showed lower O2 consumption and heat production than wildtype mice, indicating lower metabolic rate in the knockout mice . There wGDF15 knockout mice showed no gross abnormalities, suggesting that GDF15 is not essential in maintaining basic life functions. When put on obesogenic high fat diet, male GDF15 knockout mice quickly gained more weight than wildtype mice and developed worsened glucose tolerance, suggesting a protective role of endogenous GDF15 in response to obesity-inducing conditions. Energy homeostasis is maintained by the balance between energy intake and energy expenditure. Compared to the wildtype control animals, GDF15 knockout mice had higher food intake and lower metabolic rate, both likely have contributed to the aggravated obesity development. Total energy expenditure is the sum of basal metabolism, food induced heat production, thermoregulation and energy expenditure associated with physical activity. Because GDF15 knockout mice had lower locomotor activity, reduction in energy expenditure associated with physical activity is likely a contributing factor to the lower metabolic rate. It however remains to be elucidated if GDF15 deficiency also affects basal metabolism and thermoregulation.In our study, female mice showed much less robust phenotype than male mice in many parameters. Female mice were reported to be significantly less sensitive than male mice to high fat diet-induced obesity, insulin resistance, systemic inflammation and learning deficits \u201331. We hAn unexpected finding we made was the increased incidences of skin lesions in the knockout DIO mice. The incidences have resulted in reduced cohort sizes in our longitudinal study because the condition is known to cause systematic pathologic changes and confound metabolic studies , 37, andS1 Fig(A) PCR analysis of ear samples from wildtype, heterozygous, homozygous knockout mice. (B) Circulating murine GDF15 levels in wildtype and knockout mice on normal chow. (C) Circulating murine GDF15 levels in wildtype and knockout mice on high fat diet.(EPS)Click here for additional data file.S1 Table(EPS)Click here for additional data file."} +{"text": "The European Centre for Disease Prevention and Control (ECDC) invites candidates to express their interest in one or more of the following services:\u00a0SUB LIST 1 Rapporteurs services;Eurosurveillance;SUB LIST 2 Editing and proofreading services for the scientific journal SUB LIST 3 Editing and proofreading services for ECDC technical and scientific reports.\u00a0The deadline expires on 7 May 2023.https://etendering.ted.europa.eu//cft/cft-display.html?cftId=5314For more information, visit the European Union (EU) institutions' eProcurement platform:"} +{"text": "Animals are thought to achieve lignocellulose digestion via symbiotic associations with gut microbes; this view leads to significant focus on bacteria and fungi for lignocellulolytic systems. The presence of biomass conversion systems hardwired into animal genomes has not yet been unequivocally demonstrated.Capsaspora owczarzaki and together with comparative analyses on 126 crustacean transcriptomes, we found that animals are living bioreactors at a microscale as they encode enzymatic suites for biomass decomposition. We identified a total of 16,723 GH homologs that are further classified into 60 GH families. Strikingly, through phylogenetic analyses, we observed that animal lignocellulosic enzymes have multiple origins, either inherited vertically over millions of years from a common ancestor or acquired more recently from non-animal organisms.We perform an exhaustive search for glycoside hydrolase (GH) genes from 21 genomes representing major bilaterian and basal metazoan (Porifera and Cnidaria) lineages. We also assessed the genome of a unicellular relative of Metazoa, We have conducted a systematic and comprehensive survey of GH genes across major animal lineages. The ability of biomass decay appears to be determined by animals\u2019 dietary strategies. Detritivores have genes that accomplish broad enzymatic functions while the number of GH families is reduced in animals that have evolved specialized diets. Animal GH candidates identified in this study will not only facilitate future functional genomics research but also provide an analysis platform to identify enzyme candidates with industrial potential.The online version of this article (10.1186/s12864-018-4861-0) contains supplementary material, which is available to authorized users. Increasing global demands for fossil fuels have led to investigations into alternative sources of renewable energy. As one of the most abundant reserves of photosynthetically fixed carbon on earth, plant lignocellulose provides a sustainable source of polysaccharides for fermentation to biofuels that may be harnessed to meet industrial and domestic needs. Acquiring simpler metabolites from lignocellulose is challenging due to the difficulties faced by enzymes in accessing the crystalline structure of cellulose that is encapsulated by lignin. Successful digestion of lignocellulosic tissues hence requires partial breakdown of lignin, which is achieved by fungi through the release of oxidizing free radicals that target woody cell wall components \u20133.Aspergillus sp. and Trichoderma reesei for commercial preparations [The traditional dogma that animals rely on endosymbionts for lignocellulose digestive capabilities because they lack endogenous cellulases has steered researchers to focus on fungi and bacteria. Industrial lignocellulosic bioprocessing mainly relies on enzymes isolated from the fungal species such as arations \u20137. Moreoarations \u201310 and marations , 12.In recent years, multiple studies have begun to shed light on the presence of endogenous lignocellulolytic systems in animals, particularly in invertebrates \u201317. One Taxonomic classification of all GH entries from the Carbohydrate-Active Enzymes (CAZy) database , 19 reveWe retrieved 188,668 lignocellulolytic glycoside hydrolase (GH) sequences from the CAZy database where thCapsaspora owczarzaki contains 45 GH genes categorised into 21 families, five of which are cellulases and the starlet sea anemone (Nematostella vectensis) respectively , 26. Morses Fig.\u00a0, suggestely Fig.\u00a0. These pely Fig.\u00a0.Fig. 2GHLottia gigantea) and eight from the Pacific oyster (Crassostrea gigas), while a significant reduction is observed in other members of Spiralia, i.e. Platyhelminthes , which point to the prospect of metazoan homologs related by vertical descent to collectively accomplish a wide range of biomass decomposition functions.The hypothesis that cellulase GHs in animals have emerged as functional life history adaptations is valid, despite their equivocal evolutionary origins. Two mechanisms can best explain the presence of animal cellulases: (1) intermittent acquisition of genes horizontally from bacteria or fungi; and (2) vertical inheritance of genes from a metazoan ancestor followed by differential gene loss in many extant taxa. Our examination of metazoan cellulase homologs revealed that GH1, GH3, GH5, GH16 and GH39 are likely derived from bacteria and/or fungi, perhaps through associations with intestinal microbiota where information on species names are available [http://bioinf.shenwei.me/taxonkit/). Sankey diagrams were generated using RAWGraphs (https://rawgraphs.io/) [The 188,668 glycoside hydrolase (GH) sequences were retrieved from the CAZy database to enablhttp://www.uniprot.org/proteomes/) and accession numbers are provided in Additional\u00a0file\u00a0https://www.ebi.ac.uk/ena) with accession numbers provided in Additional\u00a0file\u00a0\u2212\u20095, best reciprocal BLAST hits against the GenBank non-redundant (nr) database and redundant transcripts having at least 98% identity were collapsed using CD-HIT (https://github.com/weizhongli/cdhit). We then utilized HMMER hmmscan employing hidden Markov models (HMM) profiles [Reference proteomes of fully sequenced genomes were obtained from Uniprot . MultiplAdditional file 1:Figure S1. Heatmap depicts 84 GH families identified from CAZy that do not have metazoan representatives. The number of genes within each GH family and taxon are color-coded according to a log10 scale. Dendrograms present clustering of taxa (columns) and GH families (rows) based on hierarchical clustering with Euclidean distance metric and average linkage. Black boxes denote absent members within a particular GH family. (PDF 143 kb)Additional file 2:Figure S2. Distribution of GH families retrieved from the CAZy database and identified in this study from metazoan genomes. (A) Pie charts represent the proportion of CAZy GH genes grouped according to taxa and GH families. (B) Proportion of CAZy GH genes within selected taxa are depicted. (C) The proportion of GH families identified from metazoan genomes are represented as pie charts grouped by species and by GH family. Numbers alongside pie charts in parentheses represent the total number of sequences. (PDF 1060 kb)Additional file 3:Figure S3. Taxonomic Sankey diagram of CAZy glycoside hydrolases from Bacteria. (PDF 2021 kb)Additional file 4:Figure S4. Taxonomic Sankey diagram of CAZy glycoside hydrolases from Archaea. (PDF 869 kb)Additional file 5:Figure S5. Taxonomic Sankey diagram of CAZy glycoside hydrolases from Fungi. (PDF 1716 kb)Additional file 6:Figure S6. Taxonomic Sankey diagram of CAZy glycoside hydrolases from Viridiplantae. (PDF 997 kb)Additional file 7:Figure S7. Taxonomic Sankey diagram of CAZy glycoside hydrolases from Metazoa. (PDF 1211 kb)Additional file 8:Table S1. List of accession numbers for species used in this study. (XLSX 23 kb)Additional file 9:Table S2. Summary of glycoside hydrolase annotations in metazoan genomes. (XLSX 86 kb)Additional file 10:Table S3. Summary of glycoside hydrolase annotations in crustacean transcriptomes. (XLSX 468 kb)Additional file 11:Figure S8. Heatmap illustrating the abundance of GH genes identified from 126 crustacean species. The number of GH genes within each family and taxon are color-coded according to a log2 scale. Dendrograms present clustering of species (rows) and GH families (columns) based on hierarchical clustering with Euclidean distance metric and average linkage. Black boxes denote absent members within a particular GH family. (PDF 441 kb)Additional file 12:Fasta file of glycoside hydrolase sequences identified from metazoan genomes. (TXT 1687 kb)Additional file 13:Fasta file of glycoside hydrolase sequences identified from crustacean transcriptomes. (TXT 25708 kb)"} +{"text": "Nuclear progesterone receptor (nPR) is an evolutionary innovation in vertebrates that mediates genomic responses to progesterone. Vertebrates also respond to progesterone via membrane progesterone receptors (mPRs) or membrane associated progesterone receptors (MAPRs) through rapid nongenomic mechanisms. Lampreys are extant agnathan vertebrates, residing at the evolutionary juncture where vertebrates diverged from invertebrates. A survey of the progesterone receptor (PR) gene sequences in lamprey genomes would inform PR gene evolutionary events during the transition from invertebrates to vertebrates.Petromyzon marinus and Lethenteron japonicum). To infer the origin and evolutionary history of PR genes, we constructed phylogenetic trees of PR homologous sequences across representative species of metazoans. Phylogenetic analyses revealed that the mPR\u03b3 gene first appeared in non-bilaterians, and the mPR\u03b2 gene likely arose from a duplication of mPR\u03b3. On the other hand, the mPR\u03b3 gene gave rise to the mPR\u03b4 and \u03b5 genes much later in the vertebrate lineage. In addition, the mPR\u03b1 gene first appeared in cartilaginous fishes, likely derived from duplication of mPR\u03b2 after the agnathan-gnathostome divergence. All known MAPR genes were present in the lamprey genomes. Progesterone receptor membrane component 1 (PGRMC1), neudesin and neuferricin genes probably evolved in parallel in non-bilaterians, whereas two copies of PGRMC genes probably derived from duplication of ancestral PGRMC1 sequence and appeared before the speciation of lampreys.In this study, we annotated sequences of one nPR, four mPR and four MAPR genes from genomes of two lamprey species contains supplementary material, which is available to authorized users. Amino acid sequences of Cephalochordata (Branchiostoma floridae) were downloaded from NCBI RefSeq (http://www.ncbi.nlm.nih.gov/refseq/). These amino acid sequences were subjected to BLASTP search against the nPR, mPR and MAPR proteins from human, mouse and zebrafish genomes. The BLAST hit proteins were then confirmed by BLASTP against NCBI nr database to ascertain that these identified proteins are actual progesterone receptors and not some members of Class II PAQR family.Deduced amino acid sequences from different metazoan taxonomies were downloaded from the Ensembl genome browser (release75), including Porifera (B. floridae), fruit fly, red flour beetle (T. castaneum) and placozoa (T. adhaerens). Protein sequences in each gene family were aligned by ClustalW2 [Representatives of nPR, mPR and MAPR protein sequences identified from aforementioned metazoan species were used for phylogenetic analyses and maximum likelihood (ML) approaches with 100 bootstrap replicates using MEGA6 software package . The NJ Protein-coding genes adjacent to mPR and MAPR genes in human, mouse, zebrafish and Florida lancelet genomes were identified from NCBI Map Viewer. The gene arrangements in lampreys were investigated with lamprey genome annotation files. The gene names of lamprey and Florida lancelet were further confirmed by BLASTP against NCBI human, mouse, zebrafish and nr databases. Five genes adjacent to mPR and MAPR genes in both directions (5\u2032 and 3\u2032) were presented and compared among human, mouse, zebrafish, lampreys and Florida lancelet.https://scansite4.mit.edu/4.0/#scanProtein) under medium and high stringency settings.Protein motifs were predicted using the Scansite cell signaling interactions prediction \u201cMotifScan\u201d module Additional file 2:Species selected for phylogenetic analyses of nPR, mPR and MAPR genes. (XLSX 20 kb)Additional file 3:Gene structure of nPR, mPR and MAPR genes in human, mouse and zebrafish. (PDF 368 kb)Additional file 4:Predicted motif sequences for lamprey PRs using the Scansite cell signaling interactions prediction \u201cMotifScan\u201d module. (XLSX 15 kb)Additional file 5:A phylogenetic tree constructed by the ML method demonstrates the evolutionary relationship among nPR, ER and ERR in metazoans. (PDF 12 kb)Additional file 6:Gene location of mPR and MAPR genes in sea lamprey and Japanese lamprey. (XLSX 30 kb)Additional file 7:Gene structure of four transcript isoforms of sea lamprey mPR\u03b2. (JPG 1194 kb)Additional file 8:Gene structure of four transcript isoforms of sea lamprey mPR\u03b4. (JPG 1112 kb)Additional file 9:A phylogenetic tree constructed by the ML method demonstrates the evolutionary relationship among five members of mPR gene family in metazoans. (PDF 16 kb)Additional file 10:Phylogenetic trees constructed by the ML method demonstrate the evolutionary relationship of PGRMC, neudesin and neuferricin in metazoans. (JPG 4300 kb)Additional file 11:Syntenic analysis of mPRs and MAPRs genomic sequences among human, mouse, zebrafish and lampreys. (PPTX 169 kb)"} +{"text": "This study examined how social engagement (SE) and mild cognitive impairment (MCI) influence changes in mobility over three years of follow-up. We performed a secondary analysis of longitudinal data among primary care patients aged >64 years (N=430). Mobility outcomes include performance-based function via the Short Physical Performance Battery (SPPB) and patient reported function via the Late-Life Function Instrument (LLFI). Independent variables include: 1) MCI determined by a comprehensive cognitive battery and scores 1.5 SD < age-adjusted mean; 2) SE measured by standardized self-report of social activities. Multivariate linear mixed regression models demonstrate that MCI is associated with lower scores on SPPB and LLFI . SE is associated with higher scores on SPPB and LLFI , and partially mediates the association between MCI and on each outcome. SE is linked to mobility decline especially among participants with MCI."} +{"text": "KY689112\u2013KY689138.In this data article, we provide five datasets of mantis mitochondrial genomes: (1) PCG123: nucleotide sequences of 13 protein-coding genes including all codon positions; (2) PCG123R: nucleotide sequences of two rRNAs and 13 protein-coding genes including all codon positions; (3) PCG12: nucleotide sequences of 13 protein-coding genes without third codon positions; (4) PCG12R: nucleotide sequences of two rRNAs and 13 protein-coding genes without third codon positions, and (5) PCGAA: amino acid sequences of 13 protein-coding genes. These were used to construct phylogenetic relationships within Mantodea and the phylogenetic trees inferred from Bayesian analysis using two data sets and Maximum Likelihood analysis using four data sets . We also provide initiation codon, termination codon, amino acid length and nucleotide diversity (Pi) of protein-coding genes among 27 mantises. The whole mitochondrial genomes of 27 praying mantises were submitted to GenBank with the accession numbers Specifications tableValue of the data\u2022The mitochondrial genomes of praying mantises are good models for future study of gene rearrangements and gene duplications.\u2022The primer strategy used to amplify the mantis mitochondrial genomes could be widely used for other insect mitochondrial genomes and this strategy can greatly reduce the experimental workload needed to acquire whole genome sequences.\u2022The phylogenetic relationships within Mantodea inferred from BI analyses using 2 data sets and ML analysis using four data sets show a few differences with the phylogenetic relationships reported in the main text, which is worthy of further discussions.\u2022The data presented here will be useful to solve the phylogenetic relationships within Mantodea.1The data presented here originate from a study of higher tRNA gene duplication in the mitogenomes of praying mantises and the phylogeny within Mantodea Five data sets were used to perform Maximum Likelihood analysis (ML) and Bayesian Inference (BI): (1) PCG123: 13 PCGs including all codon positions; (2) PCG123R: two rRNAs and 13 PCGs including all codon positions; (3) PCG12: 13 PCGs without third codon positions; (4) PCG12R: two rRNAs and 13 PCGs without third codon positions, and (5) PCGAA: amino acid sequences of 13 PCGs. The phylogenetic relationships inferred from BI analyses using 3 data sets and ML analyses using the data set PCG123R shared the same topologies. Hence, we illustrated nodal supports from the four analyses together, which are data provided in the main text 2Our routine experimental approach was as follows: acquisition of the whole mitochondrial genomes of 27 mantises using total DNA extraction, PCR and sequencing; sequence analyses including assembly, annotation and alignment; and construction of phylogenetic relationships. The primer strategy is shown in"} +{"text": "Romantic partners exhibit dyadic covariation (synchrony) in physiological parameters. This study aims to link everyday cortisol synchrony to daily partner interactions and empathy. We conducted coordinated multilevel analysis using data from two independently collected samples of older couples who completed questionnaires and provided salivary cortisol samples 5 to 7 times daily for 7 days. Cortisol levels were significantly correlated among partners in both studies. Cortisol synchrony was higher when partners were present (Study 1), and when partner interactions involved feeling understood and valued (Study 1) and seeking help or closeness (Study 2). Higher cortisol synchrony was further related to greater empathic accuracy (Study 1) and greater empathy (Study 2). Thus, social bonding processes and the ability to consider other\u2019s thoughts and feelings may be intertwined with physiological synchrony in everyday life."} +{"text": "Dengue virus has recently reemerged in the southern Indian Ocean islands, causing outbreaks in Reunion Island and the Seychelles. In the present study, we determined the complete genome sequences of closely related clinical isolates of dengue virus type 2 circulating in the Seychelles in 2016 and Reunion Island in 2018. Dengue virus has recently reemerged in the southern Indian Ocean islands, causing outbreaks in Reunion Island and the Seychelles. In the present study, we determined the complete genome sequences of closely related clinical isolates of dengue virus type 2 circulating in the Seychelles in 2016 and Reunion Island in 2018. Flavivirus genus (Flaviviridae family), has reemerged, causing outbreaks in Reunion Island and the Seychelles. Between December 2017 and April 2019, 49,000 DENV-2-infected cases with 428 hospitalizations were recorded in Reunion Island using Bowtie2 v2.1.0 and without recent travel history. RNA was extracted using the QIAamp kit (Qiagen). cDNA was synthesized using ProtoScript II with random primers (New England) and was purified using the QIAquick kit (Qiagen). Libraries were prepared with the Nextera XT kit (Illumina) and subjected to paired-end sequencing (2 \u00d7 250\u2009bp) on an Illumina HiSeq 4000 system (Genoscreen). The total read counts were approximately 1,034,620 (RUJul) and 622,542 (RU16417). The read quality was assessed by FastQC (KX380828). Phylogenetic analysis of the two genomes with full sequences available for representative DENV strains (UTRs of 93 nucleotides (5\u02b9 UTR) and 452 nucleotides (3\u02b9 UTR) were manually added, and the resulting genomic sequences of about 10,724 nucleotides were aligned with a DENV reference set and annotated using Geneious and MN272405 (RU16417). Raw data are available in the NCBI SRA under BioProject accession number PRJNA575805.The assembled genomes were deposited in GenBank under accession numbers"} +{"text": "QO2 decreased in CL and UUO in all states using substrates for complex II, whereas it was affected only in UUO when substrates for complex I were used. Progressive decrease in mitochondrial ROS formation\u2013in the forward and reverse pathway at complex I\u2013correlates well with the inhibition of QO2 and, therefore, with decreased electron transfer at the level of complexes upstream of cytochrome c oxidase. CL and UUO transmembrane potential responses to ADP were impaired with succinate. Intense Ca2+-induced swelling was elicited in CL and UUO mitochondria. Important and selective differences exist in CL antioxidant enzymes with respect to either Sham or UUO kidneys: CL kidneys had increased mitochondrial glutathione peroxidase and cytosolic catalase activities, indicative of compensatory responses in the face of an early altered ROS homeostasis , and of a significant tendency to apoptosis. In CL and UUO, upregulation of nuclear (erythroid-derived 2)-like 2 transcription factor (Nrf2), as well as of cytoplasmic and nuclear Kelch-like ECH-associated protein 1 (Keap1) in opposition to decreased heme oxygenase-1 (HO-1), suggest impairment of the Nrf2/Keap1/HO-1 system. It is concluded that chronic obstruction impairs mitochondrial function in CL and UUO, preferentially affecting complex II.In unilateral ureteral obstruction (UUO), both oxidative stress and mitochondrial dysfunction are related to cell death. The aim of this study has been to characterize profiles of enzyme antioxidant activities and mitochondrial functioning of the contralateral (CL) compared to UUO and Sham kidneys of Balb/c mice. Kidneys were resected 14 days after obstruction for immunohistochemical and cortical mitochondrial functioning assays. Antioxidant enzymes activities were investigated in mitochondria and cytosol. Oxygen consumption (QO Mitochondrial dysfunction participates in the initiation and progression of acute kidney injury (AKI) to chronic kidney disease (CKD) , 2. Expeth day after UUO decay is the proton motive force (\u0394\u03c1) generated across the inner mitochondrial membrane during electron flux along the mitochondrial complexes [m was similar in the 3 groups [2 led to a progressive decrease of absorbance in the Sham group, which was less accentuated in the CL and UUO groups. There were no differences between CL and UUO in all range of Ca2+ concentrations.In isolated mitochondria, opening of the permeability transition pore (mPTP) leads to mitochondrial swelling . Ca2+-in density . In the rometry) , absorbaWe measured the activities of glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase in mitochondria and cytosol from the cortical region . GPx actAfter mPTP opening, cytochrome c is released, activating caspase 3 and promoting cell death . ImmunohCoenzyme Q is essential in shuttling electrons among different mitochondrial complexes , also beUUO kidneys had the UUO kidneys had a diffuse intense cytoplasmic and nuclear reactivity , Sham kiRepresentative immunostainings of caspase 3 and active caspase 3 in the cytosol and nuclei from cortical tubular cells are presented in in vitro with raising Ca2+ concentrations.The results demonstrate profound functional mitochondrial alterations in the CL kidney of mice when the UUO kidney has tissue alterations similar to those found in CKD . The con2in vitro allowed determination of the degree of mitochondrial dysfunction in the different respiratory states. Isolated cortical CL and UUO mitochondria had striking functional alterations that cannot be correlated, respectively, to humoral stimuli for compensatory growth in the case of the remaining kidney after uninefrectomy [2 in all states with succinate in CL and UUO, and only in UUO when pyruvate and malate are oxidized compared to Sham mice. QOfrectomy or with oxidized and 4, a ~10 \u03bcM; , point tm generated after mitochondrial energization in CL and UUO kidneys is notably similar to that encountered in Sham kidneys, although the delay in its formation and the loss in the response of \u0394\u03c8m to ADP in the UUO group suggests a major impairment of proton leak [With respect specifically to respiration, the profiles seen in Sham, CL and UUO mitochondria after QO present , whereas present . Moreoveton leak , 46 that2 with succinate as the respiratory substrate , CL , and UUO animals. (A) Low magnification photomicrograph of a sham operated kidney showing normal aspect. HE staining; calibration bar: 500 \u03bcm. (B) PAS staining of the Sham kidney section. Tubular basement membranes delimit the interstitial space; calibration bar: 100 \u03bcm. (C) Low magnification of the CL kidney histological section. HE staining; calibration bar: 500 \u03bcm. (D) PAS staining of the CL kidney section shows the enlargement of the interstitial space (arrow head); calibration bar: 100 \u03bcm. (E) Photomicrograph of UUO kidney section. Notice the presence of dilated tubules in both cortex and medulla. HE staining; calibration bar: 500 \u03bcm. (F) PAS staining of the UUO kidney section showing dilated tubules (arrow heads); calibration bar: 100 \u03bcm. This figure presents the structural differences among Sham Click here for additional data file.S2 FigA) Sham kidney (cortex) histological section shows tubular profiles without apoptotic tubular cells. (B) CL kidney (cortex) showing tubular profiles with an apoptotic cell (arrow). (C) UUO section (cortex) with various tubular apoptotic cells (arrows). Counterstain: 0.001% Evans blue (red), apoptotic nuclei (green), non-apoptotic nuclei DAPI (blue); calibration bar: 25 \u03bcm. (D) Kidney from Sham animal without PCNA positive tubular cell nuclei. (E) CL kidney section showing a few PCNA+ tubular cell nuclei (arrows). (F) Kidney section of UUO animals presenting some PCNA+ tubular and interstitial cells (arrows); calibration bars: 50 \u03bcm. (G) Percentage of apoptotic tubular cells in the 3 groups. (H) Percentage of PCNA+ cells. Data represent mean \u00b1 SEM (n = 6), submitted to one-way ANOVA test followed by Tukey\u2019s test. **p<0.01, ****p<0.0001. Apoptosis and proliferation were barely detected in Sham kidney. CL kidney presented with increased tubular cells apoptosis and tubular cell proliferation, while in the UUO kidney there was a more accentuated increase in both the number of apoptotic and PCNA+ tubular cells.Detection of apoptosis was performed with ApopTag fluorescein in situ apoptosis detection kit following the instructions of the manufacturer. Proliferating cell nuclear antigen (PCNA) labeling index and apoptosis labeling index represents the percentage of tubular cell nuclei reactive to PCNA or ApopTag in the total number of tubular cells in the histological field. ((TIF)Click here for additional data file.S1 File(DOCX)Click here for additional data file.S1 Table(DOCX)Click here for additional data file."} +{"text": "Conformational intrinsic disorder is a feature present in many virus proteins. Intrinsically disordered regions (IDRs) have weaker structural requirement than ordered regions and mutations in IDRs could have a lower impact on the virus fitness. This could favor its exploration of adaptive solutions. The potyviral protein VPg contains IDRs with determinants for adaptation to its host plant. To experimentally assess whether IDRs are more resistant to mutations than ordered regions, the biologically relevant interaction between mutant libraries of both VPg and the eukaryotic translation initiation factor 4E (eIF4E) and their respective wild type partner was examined using yeast two hybrid assay. Our data shows that VPg is significantly more robust to mutations than eIF4E and as such belongs to a particular class of intrinsically disordered proteins. This result is discussed from the standpoint of IDRs involvement in the virus adaptive processes. We cE.Coli produced an average of 3x104 individual clones. The higher cloning-mating efficiency in yeast (>106) insured that the original library complexity was conserved during the transfer between bacteria and yeast of the libraries during their transfer from bacteria to yeasts, a cloning strategy was optimized. We used high efficiency Gateway recombination system, bacteria strains with maximum transformation efficiency and yeast mating that typically insures a better rate of cotransformed yeast. Corresponding mating efficiencies in yeast are provided in Capsicum annuum Yolo Wonder, a bell pepper inbred line susceptible to PVY (GenBank accession AAN74644-1).Viral genome linked protein (VPg) was from a variant of PVY isolate SON41 (GenBank accession AJ 439544\u20132). The EukCapsicum annuum cv. Yolo Wonder were cloned into pDONR201 entry vector Gateway system (ThermoFisher). GeneMorph II Random Mutagenesis Kit (Agilent) was used to perform random error-prone PCR (epPCR) mutagenesis. Several DNA target amounts were tested to produce various rates of mutations ((5'TCGCGTTAACGCTAGCATGGATCTC and 5'GTAACATCAGAGATTTTGAGACAC respectively) that hybridize upstream and downstream of attL Gateway recombination sites. PCR products were loaded on agarose gel 1%. Amplicons of the expected size were extracted from the gel using NucleoSpin Gel and PCR Clean-up kit (Macherey-Nagel). AttL-flanked mutated eIF4E and VPg genes were inserted into Yeast Two Hybrid pDEST-GADT7 (activation domain fusion vector\u2014TAIR resource accession #1010229695) and pDEST-GBKT7 (DNA-binding domain fusion vector\u2014TAIR resource accession #1010229688) respectively. The eIF4E auto-activation prevented us from using the \"eIF4E-binding domain\" construct. LR recombination reactions were performed at 25\u00b0C overnight using LR Gateway LR Clonase II Enzyme mix following manufacturer recommendations. Proteinase K treatment were applied by adding 1\u03bcl of proteinase K and incubation at 37\u00b0C for 1 minute. LR products were used to transform Escherichia coli DH10B MAX-efficiency competent cells (NEB) by electroporation. Transformation products were spread and transformed cells selected on LB plates supplemented with the suitable antibiotics (Kanamycin or Ampicillin). To characterize the libraries, 32 colonies per library were sequenced using the attB1 primer (Gateway). Each resulting sequences were aligned with the WT sequence of the corresponding gene to identify mutations. Total number of mutations, number of non-synonymous, synonymous, STOP and INDEL (insertion-deletion) mutations and their respective position in the protein sequence are displayed in Genes encoding Viral protein genome-linked protein (VPg) from Potato virus Y (PVY) SON41g and eIF4E factor from utations . In case8 cells/ml) was combined with 1ml aliquot of the mutant library strain (> 107 cells/ml). Diploid yeasts were selected on a non-interaction selective medium , the number of colonies represented the whole population of variants. Yeast colonies able to grow on stringent selecting medium for interaction representing the remaining functional variants were counted on 3 to 5 independent plates. For each experiment, mating efficiencies i.e., % diploid) calculated as the percentages of cfu/ml of diploids (number of cfu on\u2013LW plates) related to cfu/ml of the lower viability partner, was > 7%. The number of clones screened, calculated as the number of cfu/ml of diploids x resuspension volume, was estimated for each mating and was > 106. Yeast colonies were counted both manually and with Open CFU, an open-source software, http://opencfu.sourceforge.net, [The Matchmaker GAL4 Two-Hybrid system 3 (Clontech) was used according to the manufacturer specifications. pDEST GADT7\u2014containing either the wt eIF4E sequence or the different mutant libraries and pDEST GBKT7- containing either the wt VPg sequence or the different mutant libraries were transformed into AH109 and Y187 yeast strains respectively, which contain two independent reporter genes HIS3 and ADE2, conferring histidine and adenine auxotrophy, respectively and driven by hybrid GAL4 promoters. Two-hybrid library screening using yeast mating were performed following the recommendations of the MatchmakerTM prefransformed libraries user manual (Clontech PT 3183\u20131) and the Make your own \u201cMate and PlateTM\u201d library system user manual (Clontech PT 4085\u20131). Each of the yeast mutant libraries \u201clow\u201d, \u201cmedium\u201d, \u201chigh\u201d were screened using yeast mating with the wild type partner: 5 ml of the wt interacting partner strain . The mutational rate was modulated by finely tune the amount of matrix. For high mutational rates, two consecutive epPCR amplifications of the starting material were performed . Pools of 32 individual clones were randomly chosen in each library and sequenced. Non synonymous mutations leading to stop codons or frame shifts were not considered. Taking into account nucleotide mutations leading to amino acid changes , the libraries were classified as \"low\", sequences containing an average of 1.5 NS mutations per kilo base (kb), \u201cmedium\u201d (3 NS mutations) or \u201chigh\u201d (7 NS mutations) .For both eIF4E and VPg coding sequences, the \u201clow\u201d, \u201cmedium\u201d and \u201chigh\u201d statistical groups were significantly different from each other .To accurately compare the mutational robustness of the two proteins, the mutation rate of eIF4E and VPg variant libraries within each of the three statistical groups has to be not significantly different , mutations were evenly distributed with a homogenous coverage along the gene sequence .4 clones. Sequencing results and their derived nucleotide substitution matrixes (Good library representativeness must satisfy a high diversity (i.e. covering a significant number of possible mutations). This corresponds to a library containing as few redundant sequences and as many full-length sequences (lacking premature termination codons) as possible. Bacteria were transformed with the six libraries and, from positive clones counting it was deduced that the libraries contained an average of 3.5 10matrixes were usematrixes . Accordi2+-eIF4E allele and VPg from PVY SON41g isolate were chosen, as Y2H test reveals their strong interaction, thereby reducing the occurrence of false positives . In order to compare the mutational robustness of both proteins, Y2H experiments were performed by parallel screening of eIF4E mutated libraries against wild type VPg and reversely, mutated VPg libraries against wild type eIF4E. For each of the three groups of libraries , the VPg and eIF4E mutant populations were compared across all mutagenesis conditions rather than specific properties of the proteins involved. This seems unlikely, as in eukaryotic proteomes, IDRs have a higher amino acid polymorphism than ordered regions , a featu\u03c1 versus the average mutation number could not be more precisely defined in order to explore a possible negative epistasis. Hence, we simply hypothesized that deleterious mutations were not interacting (no epistasis). In such a case, the VPg-eIF4E interaction, expressed as the survival rate, was expected to decline mono-exponentially. It was observed that the functional loss of the two proteins decreased with significantly different rates, which is indicative of a higher mutational robustness of VPg than eIF4E, its more structured partner values than ordered ones, that indicates a tendency of intrinsically disordered domains to evolve faster than more structured regions during potyvirus evolution . Such wein planta that a modulation of the VPg disorder content impacts the success of the virus to overcome the host resistance [in vivo on large populations of host plants (1000 and more). Notably, a comparative analysis in planta of the impact on infection phenotype of mutations introduced either in ordered or disordered regions within the virus genome will allow the assessment of IDRs mutational robustness in a true biological context and ultimately, to better understand how viruses cope with mutations.The experimental analysis of intrinsically disordered protein mutational robustness is still scarcely documented. Recently, the demonstration that the C-terminal IDR of Nodamura Virus polymerase can accommodate very diverse amino acid sequences without losing its function suggests that IDRs can be reservoirs for evolutionary innovations towards virus adaptation to environment changes . This hysistance . The queS1 Fig(PDF)Click here for additional data file.S2 Fig(PDF)Click here for additional data file.S1 Table(PDF)Click here for additional data file.S2 Table(PDF)Click here for additional data file.S3 Table(PDF)Click here for additional data file.S4 Table(PDF)Click here for additional data file.S1 Code(PDF)Click here for additional data file."} +{"text": "Enhancing dementia care is a public health priority and supporting family caregivers of persons living with dementia (PLWD) is a critical need. This poster reports the relationships between the types of care challenges reported by family caregivers and their scores on psychosocial measures. Family caregivers (N=83) participating in the FamTechCare clinical trial identified three top priority care challenges and completed a series of measures at baseline. Priority care challenges were classified using the 10-category Technology-supported Dementia Care Typology. Three of the categories were reported by an adequate number of caregivers in order to test relationships with psychosocial measures using the Kruskal-Wallis Test. Caregivers reporting 2 or 3 BPSD challenges had higher burden (p=.007), more depression (p=.022) and worse sleep quality (p=.020) compared to those reporting 0 or 1 care challenges related to BPSD. In comparison, caregivers with 2 or 3 challenges related to DE had less burden (p=.008), less depression (p=.030), and better sleep quality (p=.042), compared to those reporting 0 or 1 challenge related to DE. Caregivers identifying 2 or 3 care challenges related to ADLs also reported higher levels of depression (p=.036). Dementia caregivers face vast caregiving responsibilities. Caregivers facing BPSD challenges report greater burden and depression. These results reinforce the need for tailored interventions to assist family caregivers in the managing varied care challenges."} +{"text": "Staphylococcus aureus in patients with GPA, and chronic nasal carriage has been linked with an increased risk of disease relapse. In this cross-sectional study, we investigated changes in the nasal microbiota including a detailed analysis of Staphylococcus spp. by shotgun metagenomics in patients with active and inactive granulomatosis with polyangiitis (GPA). Shotgun metagenomic sequence data were also used to identify protein-encoding genes within the SEED database, and the abundance of proteins then correlated with the presence of bacterial species on an annotated heatmap.Ear, nose and throat involvement in granulomatosis with polyangiitis (GPA) is frequently the initial disease manifestation. Previous investigations have observed a higher prevalence of S. aureus in the nose as assessed by culture was more frequently detected in patients with active GPA (66.7%) compared with inactive GPA (34.1%). Beta diversity analysis of nasal microbiota by bacterial 16S rRNA profiling revealed a different composition between GPA patients and healthy controls (P\u2009=\u20090.039). Beta diversity analysis of shotgun metagenomic sequence data for Staphylococcus spp. revealed a different composition between active GPA patients and healthy controls and disease controls , and between healthy controls and inactive GPA patients and household controls . Patients with active GPA had a higher abundance of S. aureus, mirroring the culture data, while healthy controls had a higher abundance of S. epidermidis. Staphylococcus pseudintermedius, generally assumed to be a pathogen of cats and dogs, showed an abundance of 13% among the Staphylococcus spp. in our cohort. During long-term follow-up of patients with inactive GPA at baseline, a higher S. aureus abundance was not associated with an increased relapse risk. Functional analyses identified ten SEED protein subsystems that differed between the groups. Most significant associations were related to chorismate synthesis and involved in the vitamin B12 pathway.The presence of S. aureus and a depletion of S. epidermidis, further demonstrating the antagonist relationships between these species. SEED functional protein subsystem analysis identified an association between the unique bacterial nasal microbiota clusters seen mainly in GPA patients and an elevated abundance of genes associated with chorismate synthesis and vitamin B12 pathways. Further studies are required to further elucidate the relationship between the biosynthesis genes and the associated bacterial species.Our data revealed a distinct dysbiosis of the nasal microbiota in GPA patients compared with disease and healthy controls. Metagenomic sequencing demonstrated that this dysbiosis in active GPA patients is manifested by increased abundance of Granulomatosis with polyangiitis is a multi-system autoimmune disorder. Disease aetiopathogenesis is considered to be multi-factorial but includes a host genetic component, epigenetic modifications and the environment , 2, withStaphylococcus aureus (60\u201370%) than the general population (20\u201330%), and the presence of persistent carriage has been associated with an increased risk of disease relapse during follow-up and active GPA patients [orange ellipse]. The two disease controls (EGPA) are within the inGPA and aGPA clusters. The overall cluster separation is not statistically different . b. The inGPA and aGPA samples were grouped into one cluster . PERMANOVA test between the three groups revealed that samples from the combined GPA patients are statistically different from the healthy control (HC) samples . (PDF 155 kb)Additional file 3: Figure S3. Taxonomic annotation of longitudinal case studies using bacterial 16S marker gene sequenced species. Bacterial diversity in six patients with follow up sampling one month (n\u2009=\u20096) and 3\u2009months later (n\u2009=\u20091) is shown. In two case studies, household controls at the initial sampling time point (n\u2009=\u20092) and one month later (n\u2009=\u20091) were also available. For comparisons, seven healthy controls are shown at the bottom of the figure. The abundance of the top 27 species with a minimum 0.1% contribution in samples from longitudinal case studies with matching species from healthy controls are shown on the x axis. (PDF 629 kb)Additional file 4: Figure S4. Heatmap analysis with annotation for disease duration in months and active GPA and inactive GPA disease stage using top 28 species with a minimum abundance of 0.5% in at least one sample in the bacterial 16S rRNA dataset. (PDF 75 kb)Additional file 5: Figure S5. Heatmap analysis with annotation for disease duration in months and active GPA, inactive GPA, and disease control using top 18 Staphylococcus species with a minimum abundance of 0.5% in at least one sample in the shotgun sequenced dataset. (PDF 83 kb)Additional file 6: Table S1. Antimicrobial susceptibility testing of S. aureus isolates. (DOCX 18 kb)Additional file 7: Table S2. S. aureus isolates and relevant metadata. (XLSX 13 kb)Additional file 8: Table S3. Relative abundance of S. aureus in the bacterial 16S dataset and in the shot gun metagenomic dataset. (DOCX 16 kb)Additional file 9: Table S4. Positive MEGAN hits for shot gun sequenced Staphylococcus species. (XLSX 10 kb)Additional file 10: Table S5. 319 SEED functional protein subsystem with minimum abundance of 0.01% in base line samples. (XLSX 181 kb)Additional file 11. Supplementary Material and Method (DOCX 92 kb)Additional file 12. Bioinformatics Analysis (DOCX 45 kb)"} +{"text": "Aspergillus, Candida, Emericella and Nakaseomyces were the different genera identified and they belonged to the fungal orders Helotiales, Eurotiales and Saccharomycetales. The data explored the presence of important fungal communities in the Arctic marine ecosystem. Metagenome data is now available at NCBI under the Sequence Read Archive (SRA) database with accession no. SRP152688.The data represents the diversity and distribution of fungal communities in Kongsfjorden, Arctic. The metagenomic DNA analysis was performed using next generation sequencing technology (Illumina MiSeq). Sequence data from amplified internal transcribed spacers (ITS) 2 region with fungal-specific primers exposed 83,417 sequences belonging to 7 operational taxonomic units (OTUs). Five of these OTUs belonged to Ascomycota, and one each to Basidiomycota and unclassified group. Aspergillus (99.57%) was the dominant genus among the 5 genera followed by unknown fungi (0.30%), Candida (0.10%), Emericella (0.03%) and Nakaseomyces. The most abundant species identified were Aspergillus versicolor, Candida humilis, Emericella nidulans, Candida glabrata and unknown spp. ). DNA from both samples was pooled together (F09) for sequencing. ITS-Amplicon sequencing was performed with Illumina MiSeq sequencing system (OmicsGen LifeSciences Pvt. Ltd.). Briefly, amplification of the ITS2 region was carried out using the forward and reverse primers with sequences \"GTGAATCATCGARTC\" and \"TCCTCCGCTTATTGAT\u2019\u2019 respectively. After validation by Agilent 2100 Bioanalyzer , DNA libraries were quantified by Qubit 2.0 Fluorometer and loaded on Illumina MiSeq instrument . Sequencing was performed using a 2\u2009\u00d7\u2009300/250 paired-end (PE) configuration. MiSeq Control Software (MCS) embedded in the MiSeq instrument conducted image analysis and base calling. ITS rRNA data analysis was performed using QIIME data analysis package Marine sediment samples collected from two different locations of Arctic Kongsfjorden \u2013 KG7 and KG5 as part of Indian Arctic Expedition 2017 were used for metagenomic DNA extraction (Power Soil"} +{"text": "Scientific Reports 10.1038/s41598-019-39251-w, published online 27 February 2019Correction to: This Article contains errors in the Results section, where:\u201cAfter model averaging, we found strong support for an effect of individual behaviour on site fidelity of rural birds and of conspecific density on site fidelity of urban and rural ones , shyer rural individuals and birds breeding at higher conspecific densities having a higher probability of changing their breeding sites between successive years than their counterparts (R2\u2009=\u20090.16). Habitat, and breeding success and productivity in the previous year received strong support to explain variability in the dispersal distance of all individuals (R2\u2009=\u20090.46), urban birds, and individuals breeding successfully or having more chicks moving less than rural, and unsuccessful owls .\u201dshould read:\u201cAfter model averaging, we found strong support for an effect of individual behaviour on site fidelity of rural birds and of conspecific density on site fidelity of urban and rural ones , shyer rural individuals and urban birds breeding at higher conspecific densities having a higher probability of changing their breeding sites between successive years than their counterparts (R2\u2009=\u20090.16). Habitat, and breeding success and productivity in the previous year received strong support to explain variability in the dispersal distance of all individuals (R2\u2009=\u20090.46), urban birds, and individuals breeding successfully or having more chicks moving less than rural, and unsuccessful owls (Table 2).\u201dAdditionally, there is an error in the Legend of Figure 1, where:\u201c(a) Proportion of burrowing owls showing site fidelity (1) or changing their breeding sites between successive years (0) in rural (grey bars) and urban (black bars) habitats. (b) For individuals changing their breeding sites, the accumulated proportion of dispersing urban (grey line) and rural (black line) individuals as a funciton of distance is also shown. The maximum dispersal distance observed is indicated by a point . Vertical dashed lines show mean distances for urban (grey line) and rural (black line) birds.\u201dshould read:\u201c(a) Proportion of burrowing owls showing site fidelity (1) or changing their breeding sites between successive years (0) in rural (grey bars) and urban (black bars) habitats. (b) For individuals changing their breeding sites, the accumulated proportion of dispersing urban (black line) and rural (grey line) individuals as a function of distance is also shown. The maximum dispersal distance observed is indicated by a point . Vertical dashed lines show mean distances for urban (black line) and rural (grey line) birds.\u201dand in the Legend of Figure 2, where:\u201c(a) Factors affecting site fidelity among rural and urban burrowing owls (estimate\u2009\u00b1\u200995% CI). Site fidelity was negatively related to individual behaviour among rural individuals (b) while it was negatively related to conspecific density (measured as aggregation) among urban ones. (c) Lines show the probability of remaining in the same breeding site for individuals with different FID and living at different conspecific densities. Dots show predicted values.\u201dshould read\u201c(a) Factors affecting site fidelity among rural and urban burrowing owls (estimate\u2009\u00b1\u200995% CI) (based on SP2). Site fidelity was negatively related to individual behaviour among rural individuals (b) while it was negatively related to conspecific density (measured as aggregation) among urban ones. (c) Lines show the probability of remaining in the same breeding site for individuals with different FID and living at different conspecific densities. Dots show predicted values.\u201d"} +{"text": "In this report the data was obtained from a prospective case-control study with a sample size of sixteen patients with exudative age related macular degeneration (AMD) due to choroidal neovascularization (CNV) and eighteen patients with polypoidal choroidal vasculopathy (PCV) and fifty controls (cataract patients without any other ocular diseases). Luminex bead based multiplex assay with a panel of 41 analytes was used to study the cytokine levels in plasma and aqueous humor. Specifications TableValue of the data\u2022Here we report the plasma and aqueous humor cytokine levels in patients with exudative AMD, PCV and controls (cataract patients without any other ocular diseases).\u2022p < 0.04).In exudative AMD patients, aqueous humor cytokines GRO, MDC and MIP-1\u03b1 were significantly higher than controls (\u2022p < 0.03).In PCV patients, aqueous humor cytokines GRO, MDC, MIP-1\u03b1, MIP-1\u03b2, IL-8, IP-10 and MCP levels were significantly higher than controls , polypoidal choroidal vasculopathy (PCV) and healthy controls .2In a prospective case-control study, sixteen patients who were diagnosed with exudative age related macular degeneration (AMD) due to choroidal neovascularization (CNV) and eighteen patients with polypoidal choroidal vasculopathy (PCV) and fifty age and gender matched cataract patients without any other ocular complications (controls) were recruited. The study was approved by the Institutional review board of Tan Tock Seng Hospital, Singapore and adhered to the tenets of the Declaration of Helsinki. Prior informed consent was obtained from all the subjects. Five milliliters of peripheral venous blood collected aseptically from 14 AMD patients, 18 PCV patients and fifty controls and approximately 200\u202f\u00b5L of aqueous humor collected from 16 AMD patients, 16 PCV patients and 41 controls using standard protocols were used for cytokine profiling Just before analysis clinical samples were thawed on ice and centrifuged for 5 minutes at 3000\u202frpm. Supernatant and plasma were separated from aqueous humor and plasma, respectively and transferred onto the ice. Twenty five microliters of each sample was analyzed for a panel of 41 cytokines by Lumin"} +{"text": "OBJECTIVES/SPECIFIC AIMS: Many CTSA programs have implemented curricula leading to clinical investigation master\u2019s degrees. Evaluation of long-term outcomes for graduates can support curriculum improvement. METHODS/STUDY POPULATION: We evaluated graduates 1\u20133 years post completion of an MS in Clinical Investigation at the University of Utah. We administered the 12-item Clinical Research Appraisal Inventory (CRAI-12) describing confidence in ability to perform research tasks; we derived 6 CRAI sub-scales. Additional questionnaire items assessed current engagement in research, including percent of effort devoted to research and level of involvement in research projects using specific research methods. RESULTS/ANTICIPATED RESULTS: Graduates reported high confidence for the CRAI domain of reporting, interpreting, and presenting and the domain of conceptualizing and collaborating (16.5\u00b12.2) on research projects; confidence was somewhat lower in the domains of planning (14.6\u00b13.3) and funding (14.9\u00b12.8) projects. Graduates\u2019 estimated current professional effort devoted to research had a median of 32%, interquartile range (IQR) 20%\u201370%; among graduates with clinical responsibilities, median effort devoted to research was 23%, IQR 15%\u201345%. In total, 74% of graduates reported moderate or high involvement in research using existing large databases, 46% reported moderate or high involvement in comparative effectiveness research, and 54% reported moderate or high involvement in quality improvement. DISCUSSION/SIGNIFICANCE OF IMPACT: A majority of clinical investigation graduates remain engaged in research but most are able to devote less than one-third of professional effort to research. Evaluation of clinical investigation graduates who have moved into their research careers can inform program directors about domains of research expertise and methodological areas that may merit additional emphasis in the curriculum."} +{"text": "Dementia is one of the most common reasons for needing a caregiver (CG). Few studies have compared dementia and non-dementia caregivers who have transitioned into family caregiving roles. Participants in the REasons for Geographic and Racial Differences in Stroke (REGARDS) study who transitioned into a significant caregiving role were recruited to participate in the Caregiving Transitions Study (CTS). Of 11,483 REGARDS participants who were not caregivers at baseline, 1229 (11%) transitioned into a family caregiving role. Eligibility criteria were met by 251 and they were enrolled along with 251 demographically-matched noncaregiving controls. Enrolled caregivers are 65% female; 36% African American; 71.8 + 8.1 years of age; caring for a spouse/partner (51%), parent (25%), or another person (24%). 47% are caring for a person with dementia. Dementia CGs provide more hours of care per day (9.3 hours versus 6.7 hours), report being under more stress and twice as much strain as non-dementia CGs . They feel more burdened by the care recipient\u2019s treatment (p=0.01) and report that the burden leads to delays in the care recipient receiving medical care (p<0.007). Dementia CGs are more than twice as likely as non-caregivers to report that their caregiving makes them worse at taking care of their own health . This prospective, population-based study confirms previous cross-sectional findings from convenience samples on the greater care burden experienced by dementia caregivers and extends this work to new measures of treatment burden and treatment delay."} +{"text": "Salmonella Agbeni clinical isolates withindistinguishable XbaI enzyme pattern (outbreak strain) by pulsed-fieldgel electrophoresis. The same Salmonella Agbeni XbaIpattern was isolated from a turtle in 2015; in a 2016 investigation involving the sameoutbreak strain, 63% of patients reported contact with turtles . Despite prohibition of sale of small turtles (shell length less <4 inches) inthe United States since 1975 , using high quality single nucleotide polymorphism (hqSNP) analysis, was performedby CDC on clinical isolates from the 2017 outbreak, the 2016 illness cluster, and theturtle isolate from 2015 to characterize genetic relatedness.A case was defined as isolation of Seventy-six cases were identified in 19 states in 2017; two thirds (67%) of patientsresided in East Coast states .Salmonella outbreaks linked toturtles (28%\u201333%) than multistate foodborne pathogenoutbreaks (27%) as well as recent"} +{"text": "Vaccination with tetanus toxoid, reduced diphtheria toxoid, and acellular pertussis (Tdap) vaccine is recommended for all pregnant women to protect infants who are too young for vaccination from severe pertussis-related outcomes recommends that pregnant women receive Tdap vaccine at the earliest opportunity, during 27\u201336 weeks\u2019 gestation of each pregnancy and the infant\u2019s outcome. Relative risks and 95% confidence intervals (CIs) were calculated to identify differences between vaccinated and unvaccinated mothers and barriers to receipt of prenatal Tdap vaccine. Because reporting of pertussis cases in infants aged <4 months is mandated in California, the follow-up interviews and data collected for this analysis were considered to be nonresearch data.Sixty-six (58%) mothers and their prenatal care providers completed the supplemental questionnaire during routine case investigations. Data on mother\u2019s insurance status, stocking status of Tdap vaccine by provider, or mother\u2019s gestational week of pregnancy (if vaccinated) were incomplete for six (9%) mothers and were excluded from relevant calculations. Twenty-six (39%) of the 66 interviewed mothers reported receiving Tdap vaccine during their pregnancy with the case infant; among these, 24 (92%) were vaccinated at their prenatal care provider\u2019s office. Prenatal care providers documented Tdap vaccine administration for 25 of the 26 mothers who reported vaccination; no information on a Tdap vaccine dose was found in the medical record or the California Immunization Registry for one mother. Among the 25 mothers with documentation of receipt of prenatal Tdap vaccine, 20 (80%) were vaccinated according to ACIP recommendations, during 27\u201336 weeks\u2019 gestation (median\u00a0=\u00a032 weeks). Five (20%) mothers received Tdap vaccine outside the recommended 27\u201336 week time frame . Among the 20 infants whose mothers were vaccinated during the recommended time frame, four were hospitalized, but none required admission to an ICU. Among the five infants whose mothers were vaccinated outside the recommended time frame, two were hospitalized, including one who required ICU admission and subsequently died.Among the 66 interviewed mothers, 40 (61%) did not receive Tdap vaccine during pregnancy and among these mothers, 20 (50%) of their infants were hospitalized, including eight (40%) who were admitted to an ICU, one of whom died. Among the 40 unvaccinated mothers, 10 (25%) did not receive a recommendation or referral off-site for vaccination from their prenatal care provider, nine (23%) were referred off-site for vaccination but did not receive Tdap vaccine, eight (20%) mothers reported refusing Tdap vaccine for personal reasons, seven (18%) were deferred for vaccination by their provider for a reason not considered by ACIP to be a contraindication , three (8%) did not receive prenatal care during 27\u201336 weeks\u2019 gestation, one (3%) reported a possible valid contraindication (adverse reaction to pertussis vaccination as a child), and no information was available for two (5%) women . SixteenAmong the 60 (91%) mothers with complete information, 19 (73%) of 26 women with private insurance and 15 (44%) of 34 women insured by Medicaid received prenatal care from a provider who stocked Tdap vaccine on-site . FourteeAmong the 61 interviewed prenatal care providers whose Tdap vaccine stocking policies were known, 34 (56%) stocked Tdap vaccine on-site. Among the 27 (44%) providers who did not stock Tdap vaccine on-site, 17 (63%) recommended Tdap vaccine during pregnancy, 16 of whom referred mothers off-site for vaccination, typically to a pharmacy, local public health department, or the mother\u2019s primary care physician. Two (13%) of the 16 mothers referred off-site were vaccinated, including one who was vaccinated at 38 weeks\u2019 gestation. In only one case was the Tdap dose documented in the mother\u2019s medical record. Among the 27 providers who did not stock Tdap vaccine on-site, the most common reasons cited for not stocking were cost (44%) and reimbursement (41%) issues.In this review of 66 mothers of infants aged <4 months who became ill with pertussis in 2016, 20 mothers (30%) received Tdap vaccine during the time frame recommended by ACIP, all of whom were vaccinated in their prenatal clinic during a routine visit. Mothers whose providers stocked Tdap on-site were more likely to be vaccinated than were those whose providers did not stock Tdap. This finding is consistent with a previous survey demonstrating that receipt of influenza vaccine by pregnant women was more likely among those whose prenatal care providers offered vaccination on-site vaccination is recommended for all pregnant women during 27\u201336 weeks\u2019 gestation to prevent infant pertussis, coverage among pregnant women is suboptimal.Among 66 interviewed mothers of infants aged <4 months with pertussis, 30% appropriately received Tdap vaccine. Women whose clinics stocked Tdap vaccine were more likely to be vaccinated. Women with Medicaid were less likely to be vaccinated than were those with private insurance, even when treated in clinics that stocked Tdap vaccine.Promoting on-site prenatal vaccination, educating providers about Tdap recommendations, and strengthening off-site referral likely will improve Tdap vaccination coverage during pregnancy."} +{"text": "Sox9-abrogated myofibroblasts identified >30% of genes regulated by SOX9 relate to the ECM. Further scrutiny of these data identified a panel of highly expressed ECM proteins, including Osteopontin (OPN), Osteoactivin (GPNMB), Fibronectin (FN1), Osteonectin (SPARC) and Vimentin (VIM) as SOX9 targets amenable to assay in patient serum. In vivo all SOX-regulated targets were increased in human disease and mouse models of fibrosis and decreased following Sox9-loss in mice with parenchymal and biliary fibrosis. In patient serum samples, SOX9-regulated ECM proteins were altered in response to fibrosis severity, whereas comparison with established clinical biomarkers demonstrated superiority for OPN and VIM at detecting early stages of fibrosis. These data support SOX9 in the mechanisms underlying fibrosis and highlight SOX9 and its downstream targets as new measures to stratify patients with liver fibrosis.Extracellular matrix (ECM) deposition and resultant scar play a major role in the pathogenesis and progression of liver fibrosis. Identifying core regulators of ECM deposition may lead to urgently needed diagnostic and therapetic strategies for the disease. The transcription factor Sex determining region Y box 9 (SOX9) is actively involved in scar formation and its prevalence in patients with liver fibrosis predicts progression. In this study, transcriptomic approaches of It is a feature of most chronic liver diseases and is characterized by progressive deposition of extracellular matrix (ECM) proteins resulting in pathological scaring and tissue dysfunction4. Although potentially reversible during early stages, a significant number of patients progress to advanced fibrosis and end-stage cirrhosis, increasing the risk of hepatocellular carcinoma (HCC)7. Identifying the extent of fibrosis and risk of progression would provide a valuable clinical tool.Liver fibrosis is increasing and a major cause of morbidity and mortality11. Consequently core regulators of ECM deposition could be exploited to identify useful targets for urgently needed diagnostic and therapetic strategies for the disease. We have previously identified the transcription factor, Sex determining region Y box 9 (SOX9), as a key factor regulating multiple components of the fibrotic ECM in liver disease whereas its prevalence in patient biopsy samples predicts progression towards cirrhosis16.Liver biopsy remains an important measure to assess fibrosis. However, several biological tests already in clinical use have taken advantage of secreted factors associated with the pathogenesis of liver fibrosis including ECM remodeling and deposition 18. Here SOX9 transcriptionally activates many cartilage-specific ECM genes such as Collagens type-2, 9, 11 and 27, Aggrecan, Matrillin-1 and Cartilage Oligomeric Protein19. Importantly, SOX9 is silenced in terminally differentiated chondrocytes prior to ossification17. This context-specific expression of SOX9 is mediated by several signaling pathway, many of which become dysregulated in fibrotic disease14. In light of this and in response to profibrotic signaling factors, SOX9 becomes expressed by the fibrogenic cell-type, the activated hepatic stellate cell (HSC), a myofibroblast that contains \u03b1-smooth muscle actin (\u03b1SMA) and regulates the production of ECM components, type 1 Collagen (COL1) and OPN16, both implicated in disease progression21, and inhibits the collagenase Matrix metalloproteinase-13 (MMP13)15. In the context of fibrosis and increased organ stiffness, studies in development, liver fibrosis and regeneration all support mechanisms involving the mechanosenstive factor Yes associated protein-1 (YAP-1) as important in the regulation of SOX924. In vivo, mice lacking SOX9 have significantly reduced scarring, improved liver function and less inflammation in models of fibrosis12. Data from multiple transgenic models to inactivate SOX9 support hepatic myofibroblasts as the causative cell-type mediating fibrosis12. In human patients with chronic liver disease, the profile and localization of SOX9 was identical to rodent suggesting the mechanisms underlying SOX9 function in fibrosis are likely to be the same12. Moreover, the extent of SOX9 in biopsies from patients with chronic liver disease correlated with fibrosis severity and accurately predicted disease progression towards cirrhosis12.These studies stem from SOX9\u2019s critical role in bone development and its requirement in chondrogenesis, whereby the initial cartilaginous skeletal elements are formed and serve as a template for endochondral bone formationSox9-abrogated and wild-type liver myofibroblasts, gene ontology and functional analysis identified >30% of genes regulated by SOX9 relate to the ECM. Further scrutiny of these data identified a panel of highly expressed ECM proteins, verified in vitro and in vivo as SOX9 targets that correlated with severity of fibrosis in patient serum samples.As an extension to these data placing SOX9 in the mechanism underlying fibrosis, we hypothesized that defining downstream SOX9 targets in liver myofibroblasts would identify ECM components amenable to assay in serum from patients with chronic liver disease. Through transcriptomic and experimental analysis of Sox9 depleted activated rat HSCs (ArHSCs) compared to control cells resulted in a total of 540 differentially regulated genes and Col116. As a result, we were interested to determine whether other matrix proteins were similarly regulated by SOX9 in these data. From 169 highly expressed genes (with \u22653800 arbitrary units of expression), 41% of transcripts encoded ribosomal proteins; further highlighting the abundant levels of the remaining genes in activated HSCs expression was significantly reduced by Sox9 knockdown , whereas all were reduced following Sox9-loss Figs\u00a0 and 6. S/\u2212) Figs\u00a0. In line/\u2212) Figs\u00a0 and 9, t/\u2212) Figs\u00a012.FigureThese data suggested the SOX9 downstream targets, OPN, VIM, SPARC, GPNMB and FN1, were secreted from activated HSCs and increased in fibrotic disease. To ascertain clinical significance, we investigated whether SOX9 regulated proteins could be used to assess fibrosis in serum samples from a well phenotyped, experimental cohort of patients with CHC (and non-infected controls). This cohort consisted of 50 patients with parallel serum and liver biopsy samples. Fibrosis was assessed by biopsies and staged using the Metavir system. Breakdown by fibrosis stage was: controls (n\u2009=\u200911), F0 (n\u2009=\u20096), F1 (n\u2009=\u200912), F2 (n\u2009=\u20099), F3 (n\u2009=\u20091) and F4/cirrhosis (n\u2009=\u200911). With only a single sample from a patient with F3 fibrosis, this was amalgamated into F4 as representative analysis of clinically advanced fibrosis.26, IS0 to IS6 (the latter representing the most severe fibrosis/cirrhosis) comparing no fibrosis , mild versus severe fibrosis . For each protein assayed, CoV was: OPN \u2013 6.66%, VIM \u2013 4.74%, SPARC 4.60%, GPNMB 6.01% and FN1\u20138.20%. A CoV below 10% is laboratory standard for accuracy and all ELISAs fulfilled this criterion. Serum concentrations from three of the five tested proteins had promising performance to predict fibrosis stage , Procollagen 3 N-peptide (P3NP), TIMP1, AST-to-Platelet Ratio Index (APRI) and the combined ELF panel were assayed and compared. All validated biomarkers/APRI values were taken at time of serum collection using unfrozen sample, in contrast to the experimental markers which were assessed in serum after going through a freeze-thaw cycle. Direct comparison of the AUROC values for each marker identified OPN and VIM as superior to APRI across all stages of fibrosis Table\u00a0. OPN and16. Moreover, in biopsy samples from patients with chronic liver disease we have established SOX9 as a predictive marker of progression toward cirrhosis12. In this study, we applied our knowledge of SOX9 in matrix regulation to identify and profile a panel of downstream targets in serum samples from patients with varying stages of chronic liver disease. Our transcriptomic analysis of Sox9 depleted HSCs revealed the extent of its role in regulating ECM components associated with fibrosis. Combined with our own data in liver fibrosis and studies in other organs29, we were able to further scrutinize these data to identify a cohort of highly expressed ECM proteins amenable to assay in patient serum samples, including OPN, VIM, SPARC, GPNMB and FN1. All targets were localized and highly expressed in activated HSCs from rodent and human. Four of the five targets had SOX9 binding sites in their promoter region with increased enrichment for SOX9, whereas VIM demonstrated comparative changes with Sox9 knockdown suggesting indirect regulation by SOX9. In vivo, all targets were reduced following Sox9 deletion in mouse models of fibrosis and localized to fibrotic regions in biopsy tissue from patients with severe fibrosis, commensurate with increased SOX9 levels.We have previously described a critical role for SOX9 in the mechanisms underlying liver fibrosis31. Despite their widespread use to distinguish early/no fibrosis from cirrhosis, classifying patients with intermediate stages of fibrosis has proved challenging. Combined with our own insight into the role of SOX9 in fibrosis, we tested the SOX9-regulated ECM proteins as novel serum markers in a well phenotyped cohort of patients with variable fibrosis. Our data indicted two SOX9-regulated ECM proteins, OPN and VIM have potential as biomarkers of liver fibrosis severity; interestingly, the data, for instance on AUROCs, were comparable to those from previous studies scrutinising other validated individual biomarkers and panels (reviewed in8). Supporting OPN and VIM as dynamic biomarkers, the distribution of both factors became significantly reduced in regions of scarring in mouse models of fibrosis resolution33. Conversely, elevations of OPN have been implicated in the progression of multiple chronic liver diseases, associated with fibrosis, including non-alcoholic steatohepatitis, alcoholic liver disease, and infection with either HCV or HBV35.Although liver biopsy remains the gold-standard to assess fibrosis in the case of diagnostic uncertainty, several non-invasive diagnostic tools now in clinical practice are based on mechanisms underlying the disease process. For example, physical approaches measuring liver stiffness, such as transient elastography (TE), take advantage of altered organ biomechanics in response to scarring; whereas many serum biomarker panels rely on detection of known ECM components indicative of the disease process36. Consistent with these findings, fibronectin isoforms are thought to precede \u03b1-SMA expression and be involved in myofibroblast activation by profibrotic TGF-\u03b238. Similar results were seen between the SOX9-regulated ECM proteins and three established clinical biomarkers (including APRI). Although TIMP1, HA and P3NP were better at identifying cirrhosis; OPN and VIM demonstrated superiority at earlier stages of fibrosis. This proof of concept diagnostic study provides encouragement for these biomarkers to be tested in larger phase 3 diagnostic studies. Importantly, a combined panel of SOX9 regulated ECM proteins may show superior performance for identifying earlier stages of fibrosis.Despite FN1 performing poorly as a biomarker for progressive fibrosis, our data in the experimental cohort complemented previous work highlighting its potential to predict early fibrosis12. Collectively these current and previous data highlight a potential clinical use for SOX9 and its downstream secreted targets as a measure to stratify patients with liver fibrosis alongside existing or emerging measures (e.g. cirrhosis risk score or liquid biomarkers)42.Overall, these data further support a critical role for SOX9 in the mechanisms underlying fibrosis and indicate the value of investigating SOX9-regulated pathways as serum biomarkers or as potential targets to reduce fibrosis and its progression to cirrhosis and HCC. Significantly, in our previous study, the utility of SOX9 detection in biopsy samples to detect severity and predict disease progression outperformed all other fibrosis risk factors12. Briefly, paired biopsy samples were obtained with informed consent and ethical approval from the Trent Cohort Study of Hepatitis C Virus (HCV) antibody\u2010positive patients from across the former UK Trent Health Region44 following selection and data collection criteria as previously described45. Patients who were receiving therapy or infected with human immunodeficiency virus were excluded. Liver biopsies were assessed blindly by an expert liver histopathologist based on the 7-point Ishak fibrosis stage, IS0 to IS6; the latter representing the most severe fibrosis/cirrhosis26. We identified a cohort of 152 biopsies classified as mild fibrosis intermediate or severe disease 12. From the same Trent HCV Cohort Study, we identified serum samples amenable to assay as a validation cohort. This consisted of 131 serum samples classified as IS0 , IS1\u20132 and IS5\u20136 .Liver biopsies tissue used in this study has been described previouslyAs proof of concept of diagnostic performance we tested serum samples in an independent, external cohort of patients with chronic HCV (treatment na\u00efve) and healthy controls (LREC no. 04/Q1701/58). All subjects were asked to follow dietary restrictions and avoid medicinal products for seven days prior to blood sampling and collection occurred between 9 am and 11am. All patients with HCV underwent a liver biopsy which was assessed by an expert liver histopathologist using the METAVIR scoring system. All biopsies were greater than 15\u2009mm in length and contained more than six portal tracts. All research was performed in accordance with relevant guidelines and regulations.23. Mice were housed and maintained, and animal experiments carried out, with approval from the University of Manchester Ethical Review Committee in accordance with UK Government Home Office regulations and its approval (licence P7FDDE62C). Mice were on a C57BL/6\u2009J background in housing with a 12-hour light dark cycle and food and water available ad libitum. RosaCreER mice46 were sourced from Jackson Laboratories. Sox9fl/fl mice were a kind gift from Professor Gerd Scherer47. To achieve inducible global Sox9 deletion, Sox9fl/fl mice were crossed with RosaCreER mice to generate RosaCreER:Sox9fl/fl animals. Genotyping, fibrosis induction and gene inactivation by tamoxifen for animals used in this study has been published12. Briefly, tamoxifen was injected i.p. to activate CreER activity and induce Sox9 deletion in ROSACreER:Sox9fl/fl animals. ROSACreER+/\u2212 and ROSACreER\u2212/\u2212 animals were injected with tamoxifen to control for any unexpected effects. 8 week carbon tetrachloride injections (CCl4) and two week bile duct ligation (BDL) were used to induced fibrosis in mice. Tissue and serum samples were collected at the end of the procedure for analysis.All animal research was performed in accordance with relevant guidelines and regulations, as described previously4 and BDL models of fibrosis in RosaCreER:Sox9fl/fl animals, there were four experimental groups. CCl4: RosaCreER\u2212/\u2212:Sox9fl/fl Olive Oil (n\u2009=\u20096); RosaCreER+/\u2212:Sox9fl/fl Olive Oil (n\u2009=\u20096), RosaCreER\u2212/\u2212:Sox9fl/fl CCl4 (n\u2009=\u20095), and RosaCreER+/\u2212:Sox9fl/fl CCl4 (n\u2009=\u20098). BDL: RosaCreER\u2212/\u2212:Sox9fl/fl Control (n\u2009=\u20095); RosaCreER+/\u2212:Sox9fl/fl Control (n\u2009=\u20095), RosaCreER\u2212/\u2212:Sox9fl/fl BDL (n\u2009=\u20097), and RosaCreER+/\u2212:Sox9fl/fl BDL (n\u2009=\u20095)12.For both CCl23. HSCs grown on chamber slides were also fixed in 4% PFA, then stored in PBS at 4\u2009\u00b0C as for previous work16. For IHC in tissue, 10\u2009mM sodium citrate (pH 6) was used for antigen retrieval, except for Osteopontin IHC which required pepsin . Antibodies used are listed in Supplementary Table\u00a023. All histological quantification and analysis was carried out blind from scanned slide images following histology as previously described12.Tissue samples were fixed in 4% paraformaldehyde (PFA) and processed for histology or immunohistochemistry (IHC) as described previously16. Gene silencing for SOX9 was carried out in culture activated HSCs using short interfering RNA (siRNA) as previously indicated. qPCR used intron spanning primers wherever possible (Supplementary Table\u00a016. Protein bands were detected with primary and secondary antibodies listed in Supplementary Table\u00a023.Primary rat and human hepatic stellate cells (rHSCs) were isolated and RNA and protein prepared as described previouslyTwo variations of the ELISA protocol were used due to the availability and applicability of antibodies. Sandwich ELISA, whereby a coating antibody was initially used to capture the antigen prior to a detecting antibody being used to detect the relative concentration by comparing to the optical density at 450\u2009nm of known standards. Where suitable capture antibodies were not available, direct ELISA was utilized and the serum sample was coated directly onto the plastic wells and a detecting antibody used to assay. A dilution series of known concentration recombinant human proteins (R&D systems) were used as standards. Accuracy of ELISAs was measured with coefficients of variation for each marker assayed and a sample was discounted if variation was >10% between replicates. Antibodies used are shown in Supplementary Table\u00a0http://ecrbrowser.dcode.org) and the MUltiple sequence Local AligNment and conservation visualization tool (MULAN: https://mulan.dcode.org/). Chromatin immunoprecipitation (ChIP) assays were performed as described previously16. Following chromatin isolation and immunoprecipitation with a SOX9 antibody , protein-DNA complexes were eluted, crosslinks reversed and protein degraded prior to DNA purification and PCR . Serum markers\u2019 ability to discriminate between stages of fibrosis was achieved by area under the receiver operator curve (AUROC) analysis with 95% confidence intervals stated.Data was analysed using the SPSS 19 software package in multiple samples after a minimum of 3 determinations. Where appropriate, data was expressed as mean\u2009\u00b1\u2009standard error of the mean (SEM). In samples with 2 groups, comparison was made using an un-paired Supplementary InformationSupplementary dataset 2Supplementary dataset 1"} +{"text": "Centella asiatica was conducted to investigate the effects of high/low light treatment experienced by parental ramets (F0 generation) on morphological and physiological properties of offspring ramets (F2 generation) as well as growth performance. Light environment experienced by parental ramets (F0 generation) significantly influenced petiole length, specific petiole length, internode length of stolon, leaf area, specific leaf area (SLA), leaf nitrogen and chlorophyll contents, potential maximum net photosynthetic rate (maxP) in offspring ramets subjected to parental or non-parental environments even after they were detached from the parental ramets. Potential maximum net photosynthetic rate (maxP) of offspring ramets (F2 generation) from parental ramets (F0 generation) subjected to low light treatment was significantly greater than that of offspring ramets (F2 generation) from parental ramets (F0 generation) subjected to high light treatment. Potential maximum net photosynthetic rate (maxP) of offspring ramets (F2 generation) subjected to parental light environment was greater than that of offspring ramets (F2 generation) subjected to non-parental light environment. The greatest biomass accumulation and total stolon length were observed in offspring ramets (F2 generation) subjected to low light treatment as parental ramets (F0 generation) experienced. When parental ramets (F0 generation) were subjected to low light treatment, biomass accumulation and total stolon length of offspring ramets (F2 generation) experiencing parental light environment were significantly greater than those of offspring ramets (F2 generation) experiencing non-parental light environment. Opposite pattern was observed in offspring ramets (F2 generation) from parental ramets subjected to high light treatment. Our work provides evidence that transgenerational plasticity through both morphological and physiological flexibility was triggered across vegetative generations for stoloniferous herb C. asiatica subjected to high/low light treatment. The transgenerational plasticity can allow offspring ramets to present adaptive phenotype early without lag time in response to the current environment. Thus, it is very important for clonal plants in adapting temporally and spatially heterogeneous habitats.Environmentally induced transgenerational plasticity can increase success of progeny and thereby be adaptive if progeny experiences the similarly parental environment. The ecological and evolutionary significance of transgenerational plasticity in plant has been studied mainly in the context of sexual generations. A pot experiment using the stoloniferous herb Senecio sp in offspring ramets subjected to parental or non-parental light environments.A greenhouse experiment was conducted to explicitly investigate effects of transgenerational plasticity across vegetative generations on morphological and physiological properties of stoloniferous herb f plants . Morpholf plants . The ramf plants . So, flef plants . As a cof plants . We predOur second hypothesis is that effects of transgenerational plasticity on growth performance are context-dependent. Then, we predicted that biomass accumulation and total stolon length of offspring ramets experiencing parental light environment significantly increased than those of offspring ramets experiencing non-parental light environment. Our third hypothesis is that offspring ramets reproduced from parental ramets subjected to low resource level environment should be favored in parental or non-parental environments. So, we predicted that whether in parental or non-parental light environment, biomass accumulation and total stolon length of offspring ramets from parental ramets subjected to low light treatment significantly increased than those of offspring ramets from parental ramets subjected to high light treatment.Centella asiatica (Umbelliferae) is a stoloniferous perennial herb, which is generally distributed in ditches, margins of ponds, lawns and roadsides. Each ramet is composed of two zygomorphic leaves with slender petiole. The axillary bud on the vertical stem may grow out and form stolon . The stolon usually take roots when in contact with moist substratum, forming a network of stolon above the ground.Centella asiatica were collected in Chengdu, Sichuan Province, China . During the experiment, fertilizer was applied to each pot once per week. Tap water was supplied to keep the substrate moist. After 4 months, offspring ramets of each original plant formed a \u201cramet bank\u201d .0 generationF August 2016, two parental ramets with similar size from each \u201cramet bank\u201d were grown into plastic pots (42 cm \u00d7 34 cm \u00d7 11 cm) respectively. We standardized size of the ramets by removing extra leaves and cutting the roots ; F0 generation high light + F1generation low light + F2 generation low light (HLL); F0 generation low light + F1 generation high light + F2 generation high light (LHH); F0 generation low light + F1 generation low light + F2 generation low light (LLL) were separated into root, leaf, petiole and stolon. Internode length of stolon and petiole length were measured by ruler. Specific internode length of stolon (internode length of stolon / dry weight) and specific petiole length (petiole length/ dry weight) were counted after drying to constant weight. Leaf area was measured according to the method described by After harvesting, offspring ramets (F2 generation) with similar size were chosen from each treatment. A fully expanded and mature leaf from each ramet was selected for photosynthetic measurement.A portable photosynthesis system GFS-3000 was used for measurement of photosynthesis during the last week of growth. Eight offspring ramets \u2013photosynthetic photon flux density (PPFD) curve] was generated according to the method described by maxP was calculated according to the n\u2013PPFDP curves which were fitted with a non-rectangular hyperbola model using the plotting software Origin (Under a CO States) :\u2205 was the apparent quantum efficiency, \ud835\udf03 was the convexity of the curve and dR was the dark respiration rate.Where The leaf for measurement of photosynthetic parameters was then finely ground to determine the nitrogen content with an elemental analyser . At the same time, the other zygomorphic leaf originating from the each ramet was selected to measure the absolute chlorophyll content using the dimethylsulphoxide (DMSO) chlorophyll extraction technique .After harvesting, dry weights of stolon, petiole, root and leaf were recorded after oven drying at 60\u00b0C until constant weight was obtained.0generation (F0), light treatment experienced by F2generation (F2) and their interaction (F0 \u00d7 F2) on morphological, leaf and photosynthetic properties of offspring ramets (F2 generation) as well as growth performance. Tukey HSD post hoc test was empolyed to compare difference among different treatments experienced by F2 generation. All analyses were conducted with SPSS 20.0 software .Prior to analysis, a square root transformation was used for total stolon length and a logarithmic transformation applied to petiole length. Two-way ANOVA was used to investigate the effects of light treatment experienced by F2 generation) were significantly affected by light treatment experienced by F0 generation (F0), light treatment experienced by F2 generation (F2) and their interaction (F0 \u00d7 F2) (Table 2 generation) were significantly affected by light treatment experienced by F0 generation (F0) and light treatment experienced by F2 generation (F2) (Table 0 generation (F0) (Table 2 generation) (Table Specific petiole length and internode length of stolon of offspring ramets (Fn) Table .2 generation) experiencing parental light environment significantly increased than those of offspring ramets (F2 generation) experiencing non-parental light environment from parental ramets subjected to high light treatment experiencing low light environment significantly increased experiencing low light treatment significantly increased than that of offspring ramets (F2 generation) experiencing high light treatment experiencing parental light environment significantly increased than that of offspring ramets (F2 generation) experiencing non-parental light environment from parental ramets subjected to low light treatment significantly decreased of offspring ramets (F2 generation) was significantly affected by light treatment experienced by F0 generation and light treatment experienced by F2 generation (Table mACC) of offspring ramets (F2 generation) was significantly affected by light treatment experienced by F0 generation, light treatment experienced by F2 generation and their interaction (F0 \u00d7 F2) (Table 0 generation) were subjected to low light treatment, area-based leaf chlorophyll content (aACC) and mass-based leaf chlorophyll content (mACC) of offspring ramets (F2 generation) experiencing parental light environment significantly increased than those of offspring ramets (F2 generation) experiencing non-parental light environment and mass-based leaf chlorophyll content (mACC) of offspring ramets (F2 generation) from parental ramets subjected to high light treatment of offspring ramets (F2 generation) was significantly affected by light treatment experienced by F0 generation, light treatment experienced by F2 generation and their interaction (F0 \u00d7 F2) (Table MN) of offspring ramets (F2 generation) was significantly affected by light treatment experienced by F2 generation (F2) (Table 0 generation) were subjected to low light treatment, leaf nitrogen content per unit area (AN) of offspring ramets (F2 generation) experiencing parental light environment was significantly greater than that of offspring ramets (F2 generation) experiencing non-parental light environment of offspring ramets (F2 generation) subjected to low light treatment was significantly greater than that of offspring ramets subjected to high light treatment of offspring ramets (F2 generation) was significantly affected by light treatment experienced by F0 generation and interaction between light treatment experienced by F0 generation and light treatment experienced by F2 generation (F0 \u00d7 F2) (Table maxP) of offspring ramets (F2 generation) from parental ramets subjected to low light treatment was greater than that of offspring ramets (F2 generation) from parental ramets subjected to high light treatment of offspring ramets (F2 generation) subjected to parental light environment was greater than that of offspring ramets subjected to non-parental light environment and light treatment experienced by F2 generation (F2) (Table 2 generation) subjected to low light treatment as parental ramets (F0 generation) experienced were subjected to low light treatment, biomass accumulation and total length of stolon of offspring ramets (F2 generation) experiencing parental light environment were significantly greater than those of offspring ramets (F2 generation) experiencing non-parental light environment from parental ramets subjected to high light treatment significantly influenced morphological and physiological properties of offspring ramets (F2 generation) as well as growth performance even after they were detached from the parental ramets. The results supported our first hypothesis that effects of transgenerational plasticity on morphological and physiological properties can transmit across vegetative generations of C. asiatica. Due to limited opportunities to adapt to environmental changes genetically, transgenerational plasticity can impose substantial impact on population dynamics were subjected to low light treatment, offspring ramets (F2 generation) experiencing parental light environment presented better growth performance than offspring ramets (F2 generation) experiencing non-parental light environment. This is consistent with previous study that if offspring ramets spread into a new environment as experienced by parental ramets, transgenerational plasticity may facilitate establishment of their populations by enabling adaptation to the new environment more rapidly than natural selection . In addition, opposite pattern was observed in offspring ramets (F2 generation) from parental ramets subjected to high light treatment. The results supported our second hypothesis that effects of transgenerational plasticity on growth performance are context-dependent. Such transgenerational plasticity spanning across vegetative generations is likely adaptive in clonal species from parental ramets (F0 generation) subjected to high light treatment, growth performance of offspring ramets (F2 generation) from parental ramets (F0 generation) subjected to low light treatment was favored in parental or non-parental light environment. The results supported our third hypothesis. Habitat-specific DNA methylation of clonal genotypes from natural populations may result in locally specialized ecotypes (Compared to offspring ramets (Fecotypes . Furtherecotypes .2-elevated environment reduced photosynthesis compared to seedlings from parents grown in ambient CO2 conditions (2 generation) experiencing parental light environment than in offspring ramtes (F2 generation) experiencing non-parental light environment when parental ramets (F0 generation) were subjected to low light treatment. We tentatively concluded that in response to low light treatment, variation of morphological and physiological properties in parental ramets was transmited to their offspring ramets (0 generation) were subjected to low light treatment, the greatest leaf nitrogen content per unit area (AN) was observed in offspring ramtes (F2 generation) experiencing parental light environment with a decrease of specific leaf area (SLA). The results implied that effects of transgenerational plasticity on photosynthesis of offspring ramets might be mediated by alternation of resources allocation toward the photosynthetic apparatus depended on environmental characteristics experienced by their parent and themselves. Clonal plants thus have the potential to reflect past and current environmental conditions even anticipate future conditions (Furthermore, clonal plants have the potential to selectively place ramets and to avoid unfavorable conditions through morphological plasticity of spacer or branching intensity . For Polironment . In addiironment . In our nditions . So, theC. asiatica subjected to high/low light treatment. Life-history traits such as clonal integration, intraclonal division of labor and clonal architecture et al may be advantageous to exploitation and colonization of clonal plants in heterogeneous habitats (Latzel and Klime\u0161ov\u00e1, 2010; To the best of our knowledge, there has been rare study directly examining the effects of transgenerational plasticity on clonal plants through both morphological and physiological properties. Our work provides evidence that transgenerational plasticity through both morphological and physiological flexibility was triggered across vegetative generations for stoloniferous herb All authors conceived, designed, and performed the experiments and wrote the paper.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Affiliation 2 for authors Elisabetta Petracci and Oriana Nanni is incomplete. The correct affiliation 2 is: Unit of Biostatistics and Clinical Trials, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy."} +{"text": "This study evaluated neuropsychiatric symptom clusters in a sample of home residing persons with dementia, and the impact of distinct clusters on family caregiver outcomes. The expanded Neuropsychiatric Inventory (NPI-C), including measures of frequency, was collected at baseline from 250 caregivers enrolled in Reducing Agitation in People With Dementia: the Customized Activity Trial [NCT01892579]. Principle component analyses were conducted resulting in an eight behavioral clusters, accounting for 44% of total variance: 1=agitation/aggression; 2=anxiety; 3=apathy/withdrawl; 4=impulsivity; 5=psychosis; 6=restlessness; 7=circadian disturbance; 8=depression. In multiple linear regressions caregiver burden was significantly influenced by the anxiety cluster. Caregiver depression was significantly influenced by apathy/withdrawal cluster, and quality of life was significantly associated with anxiety and circadian disturbance clusters. Dimensional representation of neuropsychiatric symptom clusters can be useful in assessing the effect of multiple co-occurring symptoms on caregiver outcomes and for planning personalized intervention strategies for persons with dementia."} +{"text": "MKK3 is a member of the dual specificity kinase group specific upstream activator of p38 MAPK proteins. We originally identified MKK3 as mutant p53 (mutp53) gain-of-function (GOF) upregulated target gene in different tumor models. To deeply investigate the MKK3 functions in cancer, taking advantage of a panel of authenticated colorectal cancer (CRC) lines and primary colonocytes, we found that MKK3 activates specifically p38delta MAPK protein, which signaling is further triggered by 5-fluorouracil (5-FU) treatments, a largely adopted chemotherapeutic drug in CRC clinical practice. The overall achieved results proposed the MKK3/p38delta MAPK as relevant molecular axis involved in abrogating efficacy to 5-FU treatments in CRC. This commentary will provide an overall discussion of the results that have been achieved contextualizing them in the overview of the knowledge in the p38 MAPK field in cancer disease. Colorectal cancer (CRC) is one of the most common malignant tumor worldwide, thus the understanding of its underlying molecular mechanisms is crucial for the development of therapeutic strategies. The Mitogen Activated Protein Kinase-Kinase 3 (MKK3) belongs to a dual specificity kinase group (MKK) and is activated by a wide array of upstream kinases MEKK1\u20134) through Ser-189 and Thr-193 phosphorylation sites. MKK3 serves, together with MKK6, as a specific activator of p38 Mitogen Activated Protein Kinase (MAPK) family members \u20134 throug, our groWe originally identified and validated MKK3 as a mutant p53 transcriptional target gene involved in the acquisition of novel oncogenic functions through which mutant p53 actively sustains tumor malignancy , suggestIn this view, taking advantage of a panel of colorectal cancer (CRC) lines, we found that int50\u2009>\u2009100\u2009nM is reported), but that would simultaneously target the p38alpha MAPK (IC50\u2009=\u20094\u2009nM) [Such observations, performed in the same CRC models, highlights that the ideal manipulation of the p38 MAPK pathway for therapeutic purposes should aim at selective inhibition of those p38 MAPK signaling arms responsible for pro-tumoral effects while leaving those responsible for anti-tumor effects unaffected. Specifically, in CRC while p38delta MAPK targeting is desired, p38alpha MAPK should remain active. However, when it comes to already available therapeutic tools, it is to be acknowledged that p38 MAPK inhibitors have been developed to target specifically the p38alpha MAPK isoform because of its wide tissue distribution and abundance. As a result, currently available p38 MAPK targeting drugs only display partial isotype selectivity making t\u2009=\u20094\u2009nM) could alOverall, the identified molecular mechanisms, involving MKK3 in supporting proliferation and survival signaling in CRC, suggest MKK3 as a novel and extremely attractive therapeutic target for the development of promising strategies for the management of CRC patients."} +{"text": "This project characterizes spread of quality improvement (QI) projects initiated in the U.S. Department of Veterans Affairs (VA) Geriatric Scholars Program (GSP) workforce development program. This mixed methods study analyzes a recent cross-sectional survey of GSP participants and program-level data on participant characteristics and QI project topics. We surveyed 578 scholars who had completed all program requirements to that point, and still worked for VA; 207 (35%) responded. The majority of respondents who had been in the program for at least six months (70%) reported sustainment of their QI project beyond initial implementation and nearly a third (30.4%) reported any spread beyond their own care team. QI project topics spanned many domains and percent of projects reporting spread varied across domains from 0% to 67%. A workforce development capstone activity in which participants demonstrate substantive and methodological competency can foster bottom-up QI activities in a large, diverse health care system."} +{"text": "EBF3 and/or TBC1D16 gene loci. To further evaluate whether these epigenetic changes may act more generally as drivers of tumour onset and metastasis, we have investigated DNA methylation changes involving EBF3 and TBC1D16 in additional publicly available data of multiple different tumour types.Characteristic DNA methylation differences have been identified between primary and metastatic melanomas at EBF3 were observed in a number of metastatic tumour types, when compared to normal or primary tumour tissues, as well as in tumour vs normal tissues and in a colorectal primary/metastasis pair, although not all tumour samples or primary/metastasis cancer pairs exhibited altered patterns of EBF3 methylation. In addition, hypomethylation of TBC1D16 was observed in multiple tumours, including a breast cancer primary/metastasis pair, and to a lesser degree in melanoma, although again not all tumours or cancer primary/metastasis pairs exhibited altered patterns of methylation.Promoter hypermethylation and gene body hypomethylation of EBF3 and TBC1D16 are relatively common tumour-associated epigenetic events in multiple tumour types, which is consistent with a potential role as more general drivers of tumour progression.These findings suggest characteristic DNA methylation changes in Cancer is a leading cause of death worldwide. Over the recent decades, many mutations have been identified that promote tumour growth . HoweverIt is now well established that epigenetic changes are associated with tumour growth . PrimaryEBF3) in metastatic melanoma cell lines compared to matched primary melanoma cell lines. Subsequent functional analyses revealed this gene has an oncogenic role. Promoter hypermethylation and elevated mRNA levels of EBF3 were validated in an independent melanoma cohort as a potential epi-driver of metastasis. Loss of TBC1D16 methylation was associated with activation of an alternative cryptic transcript, TBC1D16-47KD, which was shown to promote melanoma proliferation and metastasis. Further in vitro and in vivo functional analyses indicated that TBC1D16-47KD promoted melanoma proliferation and metastasis, possibly by regulating Rab GTPases and EGFR activation. Hypomethylation of TBC1D16 was also shown to increase sensitivity to BRAF and MEK inhibitors but predicted poorer clinical outcome for melanoma patients [Recent genome-wide tumour studies have used comparative analysis of paired primary and metastatic tumour samples to identify epi-drivers of cancer metastasis , 10. Of e et al. , 12) ande et al. . In anote et al. analysedpatients .EBF3 showed significant loss of methylation in metastases compared to benign nevi, and one of these CpG sites showed the same degree of hypomethylation in metastases compared to primary melanoma. Furthermore, they identified three CpGs in TBC1D16 that showed a significant decrease in methylation in metastases compared to primary melanoma. Two of these CpG sites were in the dataset originally identified by Vizoso et al. [A recent genome-wide study by Wouters et al. identifiEBF3 was significantly hypermethylated in metastatic cell lines compared to matched primary cell lines [EBF3 and TBC1D16 in publicly available data of multiple different tumour types, so as to further evaluate the potential role of these two genes associated with tumourigenesis and metastasis.In previous genome-wide DNA methylation analysis of three cutaneous primary and metastatic melanoma cell line pairs using reduced representation bisulfite sequencing (RRBS), we showed that an RRBS fragment in the promoter of ll lines . In the EBF3 and in the gene body of TBC1D16, which contains a promoter for the cryptic transcript TBC1D16-47KD, as summarised in the gene maps in Fig.\u00a0EBF3 CpG sites , in a cohort of 450k data of melanocytes (n\u2009=\u20093), primary melanomas (n\u2009=\u20094) and melanoma metastases in melanoma metastases (green and magenta boxplots) compared to primary melanomas (red boxplots) or melanocytes (blue boxplots).We analysed CpG methylation in the promoter and gene body regions of TBC1D16 CpG sites in the same melanoma dataset in metastatic melanomas (green and magenta boxplots) compared to primary tumours (red boxplots). In general, melanocytes were also hypomethylated in TBC1D16. We next evaluated TBC1D16 methylation in an RRBS fragment that overlapped one of the CpG sites (cg17295878) in a series of metastatic melanoma cell lines and TBC1D16 (cg07618085) were significantly differentially methylated in lymph node metastases compared to primary tumours (green vs red boxplots). A similar significant loss of methylation was also observed in the TBC1D16-47KD cryptic promoter (cg23651872). One of the EBF3 CpG sites (cg25866634) also gained methylation (+\u200917%) in abdominal metastases (black vs red boxplots). Furthermore, three EBF3 gene body CpG sites showed a 19% gain of methylation in endometrial hyperplasia compared to primary endometrial tumours (blue vs red boxplots). In prostate cancer metastases gene body CpG and four TBC1D16 CpG sites (both +\u200916%) were observed compared to normal prostate tissue (red vs blue boxplots). For triple-negative breast cancer samples in the EBF3 gene body showed 12% loss of methylation in primary tumours compared to normal tissue (red vs blue boxplots).In endometrial cancer Fig.\u00a0a, CpG siEBF3 and TBC1D16 in 450k methylation array in both adenomas and carcinomas compared to normal colon tissue . In liver metastases, there was further 15% loss of methylation at several EBF3 gene body sites compared to carcinomas (magenta vs green boxplots), and 19% loss of methylation compared to adenomas in aberrant crypt foci compared to normal colonic crypt (green vs red boxplots). There were also four CpG sites in the EBF3 promoter that were hypermethylated +\u200930% in primary tumours of the colon compared to normal colon data [EBF3 promoter CpG sites were strikingly completely unmethylated in the EBF3 promoter compared to the primary colon cancer tumour sample, and a corresponding decrease in methylation (~\u200920%) at most sites within the EBF3 gene body in the metastatic vs primary colorectal cancer samples. With the exception of the normal brain tissue and the metastatic breast cancer cell line, TBC1D16 was largely methylated (median\u2009=\u20090.94) in all of the cell lines and samples. In the breast cancer cell lines, TBC1D16 methylation showed high discrimination between the primary (median\u2009=\u20090.96) and the metastatic (median\u2009=\u20090) breast cancer cell line pair data derived EBF3 and TBC1D16, we also investigated the same CpG sites in additional WGBS of cancer samples from TCGA, consisting of primary tumour samples for urothelial bladder carcinoma, breast invasive carcinoma, colon adenocarcinoma, glioblastoma multiforme, lung adenocarcinoma, lung squamous cell carcinoma, rectum adenocarcinoma, stomach adenocarcinoma and uterine corpus endometrial carcinoma . TBC1D16 methylation was high on average in all of the tumour samples (median\u2009=\u20090.86) except the glioblastoma multiforme sample (median\u2009=\u20090.14).To further assess the methylation of oma Fig.\u00a0. All of EBF3 and the gene body of TBC1D16, which includes a cryptic promoter for TBC1D16-47KD. This investigation was performed on multiple tumour types in comparison to normal tissues, in matched metastatic vs primary tumour tissues or cell lines and in unmatched primary tumour and normal tissues and cell lines. Differences in methylation in the promoters and gene bodies were more pronounced in higher tumour grades or in metastatic tumour tissues and cell lines vs primary tumour tissues and cell lines.Here we have investigated CpG methylation alterations in the promoter and gene body of EBF3 gene body was observed in metastatic melanoma and colorectal cancer vs primary, in both tumour tissues and cell lines. Hypomethylation in the EBF3 gene body was also observed in squamous cell carcinoma of the lung, adenocarcinoma of the lung, glioma and in colorectal cancer tissue or cell lines in comparison to normal colon tissues or cell lines. Conversely, EBF3 promoter hypermethylation was also observed in metastatic or hyperplastic cancers vs primary cancers, in comparison to normal tissues of the aforementioned tumour and cell line types. Increased methylation of a CpG site in the EBF3 promoter in primary and metastatic tumours , in metastatic melanoma compared to primary melanoma and also hypomethylation (94%) in metastatic vs primary paired breast cancer samples). In both prostate and colorectal cancers, we observed hypermethylation of TBC1D16 compared to normal prostate tissues and normal colon tissues, respectively. Hypomethylation of TBC1D16 was previously reported in metastatic vs primary melanoma, breast cancer and other tumour types [TBC1D16 were observed in a colorectal cancer primary/metastasis pair. Methylation differences in TBC1D16 have previously been reported by Vizoso et al. [TBC1D16 are a potential epigenetic driver of tumour metastasis.We also observed characteristic ur types , 13. Howo et al. . Similaro et al. , 12 and o et al. , which sEBF3, or TBC1D16, were not identified in all tumour samples, or in all primary/metastasis cancer pairs. For example, no clear overall pattern of EBF3 methylation patterns was identified between a paired breast cancer primary and metastases, although some individual sites showed differences (e.g. the CpG site at chr10:131763531 was 46% less methylated in metastasis). In addition, the direction of methylation changes in EBF3 and TBC1D16 was not necessarily conserved across different cancer types, since endometrial cancer and prostate cancer in our data showed opposite patterns of methylation changes in EBF3 gene body and promoter regions compared to melanoma and colorectal cancer.However, epigenetic changes in EBF3 and TBC1D16 were associated with tumour metastasis, which is responsible for the majority of cancer-related deaths. As a first step of metastasis, tumour cells are released from a solid tumour and circulate in the bloodstream of patients. Molecular signatures of these circulating tumour cells (CTCs), such as the methylation status of EBF3 and TBC1D16, could potentially be used as a biomarker to help determine the prognosis of metastatic cancers. Furthermore, if the identified methylation changes are causal, they may have significant relevance to early diagnosis of cancer and possibly as a therapeutic target. For example, it appears that the EBF3 promoter hypermethylation identified by RRBS [We show here that in several cancer types, characteristic methylation changes in both by RRBS may poteEBF3 promoter hypermethylation identified by RRBS [TBC1D16 CpG sites identified by 450k were not detected by RRBS due to the lack of MspI fragments encompassing the majority of those specific loci. Further sequencing-based studies, such as WGBS, will provide a more comprehensive view of the cancer methylome and will help to identify greater numbers of epigenetic markers that distinguish between primary and metastatic tumour pairs.We acknowledge that a full interpretation of the methylation results presented here should take into consideration different analysis platforms, each having their own strengths and weaknesses. For instance, in contrast to WGBS, where all CpGs in the genome were analysed, RRBS enriched for CpG-dense regions and involved sequencing of four million CpG sites but only 13.4% of the genome . On the by RRBS was not EBF3 and TBC1D16, similar to those reported previously [EBF3 and TBC1D16 are potential epi-drivers of aggressive tumourigenic changes in multiple cancer types.The present findings suggest that methylation changes in eviously , 10, arehttps://www.ncbi.nlm.nih.gov/geo/): (1) 16 melanoma metastases to the brain, 17 lymph node melanoma metastases, 4 primary melanoma tumours and 3 normal melanocyte samples (accession number GSE44661[All 450k methylation data were obtained from the NCBI gene expression omnibus (GEO) database of four normal colon, eight normal colonic crypt, nine9 aberrant crypt foci and ten primary colorectal cancer tumours.Reduced representation bisulfite sequencing (RRBS) libraries for the 12 cell lines described here WGBS data for five normal tissues/cells , six primary tumours/cells and two metastases were obtained from GEO . The bigWig files were converted to BedGraph format using bigWigToBedGraph (http://hgdownload.cse.ucsc.edu/admin/exe/linux.x86_64/).(2) TCGA WGBS data for seven urothelial bladder carcinoma (TCGA-BLCA), five breast invasive carcinoma (TCGA-BRCA), three colon adenocarcinoma (TCGA-COAD), six glioblastoma multiforme (TCGA-GBM), six lung adenocarcinoma (TCGA-LUAD), five lung squamous cell carcinoma (TCGA-LUSC), three rectum adenocarcinoma (TCGA-READ), five stomach adenocarcinoma (TCGA-STAD) and six uterine corpus endometrial carcinoma (TCGA-UCEC) were obtained from https://bedtools.readthedocs.io/) was used to extract methylation beta values for all analysed loci. Datasets were collated and non-parametric Mann-Whitney U statistical tests were performed in the R Studio environment (version 3.1.1).The BEDTools suite ("} +{"text": "In pediatric patients with deep venous thrombosis (DVT) and pulmonary embolism (PE), antiphospholipid syndrome (APS) should be considered early and efforts must be made to ensure timely diagnosis of this potentially life-threatening condition. Pediatric APS is an autoimmune disease characterized by vascular thrombosis and persistently positive antiphospholipid antibodies ,2,3,4,5."} +{"text": "Despite strong vetting for disease activity, only 10% of candidate new molecular entities in early stage clinical trials are eventually approved. Analyzing historical pipeline data, Nelson et al. 2015 (Nat. Genet.) concluded pipeline drug targets with human genetic evidence of disease association are twice as likely to lead to approved drugs. Taking advantage of recent clinical development advances and rapid growth in GWAS datasets, we extend the original work using updated data, test whether genetic evidence predicts future successes and introduce statistical models adjusting for target and indication-level properties. Our work confirms drugs with genetically supported targets were more likely to be successful in Phases II and III. When causal genes are clear (Mendelian traits and GWAS associations linked to coding variants), we find the use of human genetic evidence increases approval by greater than two-fold, and, for Mendelian associations, the positive association holds prospectively. Our findings suggest investments into genomics and genetics are likely to be beneficial to companies deploying this strategy. The growth of human genetics resources has the potential to help us develop better drugs. By looking at whether and how historical drug approvals could have been predicted from our current knowledge of human genetics, we can validate this approach and assess which types of genetic evidence are most likely to be useful in guiding drug discovery. Validation is important because we are often uncertain about the biological mechanisms behind genetic variants linked to disease. Most associated variants do not occur within protein-coding regions of the genome, and it is difficult to tell which of many nearby genes is contributing to disease risk. In this paper, we confirm previous correlations between genetic evidence and historical drug approvals. We find genetic evidence from severe genetic disorders and from genetic variants that alter protein sequence is more strongly associated with historical approvals. We offer statistical approaches for prioritizing new drug candidates based on whether their mechanisms are supported by human genetic evidence. The cost of developing new molecular entities (NMEs) into approved therapies continues to increase with cost per launched NME ranging from $3 billion to more than $10 billion across major research based pharmaceutical companies . Thus, tAnalyzing historic data of the progress of drug compounds through the drug development pipeline, Nelson et al. 2015 concludePCSK9 . Pr. Prp = 2IS score . DetailsStan . We. We24]. https://github.com/AbbVie-ComputationalGenomics/genetic-evidence-approval). The git repository also contains instructions for running a Shiny app displaying model predictions.Code and data tables required to reproduce the main text figures are provided on Github (S1 Text(PDF)Click here for additional data file.S2 Text(PDF)Click here for additional data file.S3 Text(PDF)Click here for additional data file.S4 Text(PDF)Click here for additional data file.S5 Text(PDF)Click here for additional data file.S1 FigReplication of Figure 2N from Nelson et al. supplementary datasets. Figure shows the enrichment of approved drug targets among genes with human genetic associations.(PDF)Click here for additional data file.S2 FigReplication of Figure 3N from Nelson et al. supplementary datasets. Figure shows the proportion of gene target-indication pairs with genetic associations for similar traits by pipeline phase and association source.(PDF)Click here for additional data file.S3 FigSensitivity of risk ratios p(approved | genetic support)/p(approved | no genetic support) and 95% confidence limits to choice of MeSH similarity cutoff. Nelson et al. value 0.7 shown in red. Results are computed from Nelson et al supplementary datasets.(PDF)Click here for additional data file.S4 FigSensitivity of risk ratios p(approved | genetic support)/p(approved | no genetic support) and 95% confidence limits to choice of minimum number of associations parameter. Nelson et al. value of 5 shown in red. Results are computed from Nelson et al supplementary datasets.(PDF)Click here for additional data file.S5 FigAssigned latest phase compared to Pharmaprojects status (unknown Pharmaprojects status categories such as No Development Reported and Suspended are combined) at the global and indication level.(PDF)Click here for additional data file.S6 FigLatest change date for Pharmaprojects drugs by development status. In panel Pharmaprojects Global Status, statuses come from the Pharmaprojects global status field. In panel Global Latest Phase, statuses are the latest global development phase assigned in this document.(PDF)Click here for additional data file.S7 FigEarliest date in the Pharmaprojects event history by status. In panel Pharmaprojects Global Status, statuses come from the Pharmaprojects global status field. In panel Global Latest Phase, statuses are the latest global development phase assigned in this document. Note 6% of compounds do not have any entries in their event history and are omitted.(PDF)Click here for additional data file.S8 FigReplication of Figure 2N from Nelson et al. supplementary genetic association dataset and updated pipeline data. Figure shows the enrichment of approved drug targets among genes with human genetic associations.(PDF)Click here for additional data file.S9 FigReplication of Figure 3N from Nelson et al. supplementary genetic association dataset and updated pipeline data. Figure shows the proportion of gene target-indication pairs with genetic associations for similar traits by pipeline phase and association source.(PDF)Click here for additional data file.S10 FigDate SNP associations appearing in the reanalysis were added to the GWAS Catalog by whether or not Nelson et al. reported the association. Line shows the date of the GWASdb version used in Nelson et al. 2013.(PDF)Click here for additional data file.S11 FigReplication of Figure 2N from updated GWAS Catalog genetic association dataset and updated pipeline data. Figure shows the enrichment of approved drug targets among genes with human genetic associations.(PDF)Click here for additional data file.S12 FigReplication of Figure 3Nb from updated GWAS Catalog genetic association dataset and updated pipeline data. Figure shows the proportion of gene target-indication pairs with genetic associations for similar traits by pipeline phase and association source.(PDF)Click here for additional data file.S13 FigMedian odds ratio for new reported case-control SNP-trait associations through time. A new reported SNP-trait association is one appearing in the GWAS Catalog for the first time, in contrast to a replicate of a previous association.(PDF)Click here for additional data file.S14 FigRelationship between effect size in the first study and effect size in the study with the largest sample size for GWAS Catalog case-control studies for associations that have been replicated.(PDF)Click here for additional data file.S15 FigMedian odds ratio for new reported case-control SNP-trait associations through time. A new reported SNP-trait association is one appearing in the GWAS Catalog for the first time, in contrast to a replicate of a previous association. Only studies with a later replicate are considered and the effect size reported is from the largest replicate. Only years with at least 20 replicated studies are shown.(PDF)Click here for additional data file.S16 FigA portion of the MeSH vocabulary used to illustrate semantic similarity. Information contents from the number of descendants are given in parentheses.(PDF)Click here for additional data file.S17 FigNelson et al. semantic similarity versus semantic similarity (average of Lin and Resnik similarities) computed in this analysis. Black points show a random sample of 50,000 trait pairs for which both similarities were available. Blue line shows smoothed relationship estimated using all possible similarity pairs. Dashed red lines show old and new cutoff values.(PDF)Click here for additional data file.S18 FigEstimated effect of genetic evidence on pipeline progression. Main text Figure 1b computed with similarity cutoff 0.7. This cutoff was originally used in Table 1N, but our main text figure uses similarity cutoff 0.73 because of systematic differences in computed similarities.(PDF)Click here for additional data file.S19 FigRisk ratio of Phase I to approval progression for gene target-indication pairs with and without genetic evidence for different values of the MeSH similarity cutoff split by whether MeSH similarity is automatically assigned, manually assigned, or using both automatic and manually assigned similarities (default). Full Data and New Genetic are as described in main text. 2013: computed from supplementary tables.(PDF)Click here for additional data file.S20 FigProportion of gene target-indication pairs approved against proportion of gene target-indication pairs with a GWAS Catalog associated target by indication class. Larger text designates indication classes with more gene target-indication pairs. Only classes with 50 or more pairs are shown.(PDF)Click here for additional data file.S21 FigProportion of gene target-indication pairs approved against proportion of gene target-indication pairs with a GWAS Catalog associated target by target class (GO terms). Larger text designates target classes with more gene target-indication pairs. Only classes with 50 or more pairs are shown.(PDF)Click here for additional data file.S22 FigProportion of approved gene target-indication pairs binned by target RVIS score percentile.(PDF)Click here for additional data file.S23 FigProportion of approved gene target-indication pairs binned by date first drug with target added to Pharmaprojects.(PDF)Click here for additional data file.S24 FigCoefficient estimates for the effect of genetically associated trait similarity on gene target-indication pair approval using different predictor subsets. See (PDF)Click here for additional data file.S25 FigEffect of including predictors on the estimated relationship between indication-trait similarity and approval. Estimated odds ratio of gene target-indication pair attaining approval, as a function of similarity between drug indication and the most similar trait associated with the target. Posterior median and pointwise 95% credible interval from Bayesian logistic regression.(PDF)Click here for additional data file.S26 FigEffect of excluding manually assigned trait similarities on estimated relationship between GWAS genetic support and approval. Estimated odds ratio of gene target-indication pair attaining approval, as a function of similarity between drug indication and the most similar trait associated with the target. The two colors correspond to estimates when using and when excluding manually assigned similarities. Posterior median and pointwise 95% credible interval from Bayesian logistic regression.(PDF)Click here for additional data file.S27 FigEstimated effect of OMIM genetic evidence on target-indication pair approval, excluding congenital diseases and indications with mapped MeSH term also mapped to an OMIM indication. Posterior median and 95% credible intervals based on 8907 target-indication pairs, compared to results from the full data with 20292 target-indication pairs.(PDF)Click here for additional data file.S1 Tablenassoc is the number of protein coding genes with genetic associations, napproved is the number of protein coding genes linked, naa is computed as the number of protein coding genes that are both the targets of approved drugs and have reported trait associations.(PDF)Click here for additional data file.S2 TableReplication of Table 1N from Nelson et al. supplementary datasets. Risk ratio p(approved | genetic support)/p(approved | no genetic support) and bootstrap 95% confidence intervals.(PDF)Click here for additional data file.S3 TableProportion of drug-indication pairs (Indication) or drugs having development status information available from each source. Event = Pharmaprojects event history, Info = Pharmaprojects clinical information fields, Country = Pharmaprojects country status, Global = Indication status inferred from global status.(PDF)Click here for additional data file.S4 TableAgreement between Pharmaprojects status and latest phase using each evidence source when both are assigned a known development status. Columns less, greater, and equal are the proportion of times in which the source implicates a latest pipeline phase less advanced than, more advanced than, or equal to that reported by Pharmaprojects. Arranged in order of decreasing agreement.(PDF)Click here for additional data file.S5 TableReplication of Table 1N from Nelson et al. supplementary genetic association dataset and updated pipeline data. Risk ratio p(approved | genetic support)/p(approved | no genetic support) and bootstrap 95% confidence intervals.(PDF)Click here for additional data file.S6 TableRisk ratio of pipeline progression from 2013 to 2018 by presence or absence of supporting genetic evidence and 2013 phase. Risk ratio and 95% confidence intervals. Last column gives the total number of gene target-indication pairs labeled with that phase in 2013 and the total number of gene target-indication pairs that progressed in development.(PDF)Click here for additional data file.S7 TableRisk ratio of pipeline progression from 2013 to 2018 by presence or absence of supporting genetic evidence. Risk ratio and 95% confidence intervals. Last column gives the total number of 2013 gene target-indication pairs included in the analysis and the total number of drugs that progressed in development. This table shows the risk ratio of progression to a higher phase or to approval from any starting phase.(PDF)Click here for additional data file.S8 TableReplication of Table 1N from Nelson et al. 2015 supplementary genetic association dataset and updated pipeline data, using only gene target-indication pairs not used in the original analysis either due to not being in the table of gene target-indication pairs or having an inactive development status. Risk ratio p(approved | genetic support)/p(approved | no genetic support) and bootstrap 95% confidence intervals.(PDF)Click here for additional data file.S9 TableReplication of Table 1N from Nelson et al. 2015 supplementary genetic association dataset and updated pipeline data, using only gene target-indication pairs not used in the original analysis due to not being in the table of gene target-indication pairs. Risk ratio p(approved | genetic support)/p(approved | no genetic support) and bootstrap 95% confidence intervals.(PDF)Click here for additional data file.S10 TableReplication of Table 1N from Nelson et al. 2015 supplementary genetic association dataset and updated pipeline data, using only gene target-indication pairs not used in the original analysis due to being assigned an inactive status. Risk ratio p(approved | genetic support)/p(approved | no genetic support) and bootstrap 95% confidence intervals.(PDF)Click here for additional data file.S11 TableRisk ratio of progression in clinical development from 2013 to 2018 by presence or absence of supporting genetic evidence. Calculations are performed on the subset of target-indication pairs without similar 2013 approved target-indication pairs, using similarity cutoff 0.73. Risk ratio and 95% confidence intervals.(PDF)Click here for additional data file.S12 TableRisk ratio of progression in clinical development from 2013 to 2018 by presence or absence of supporting genetic evidence. Calculations are performed on the subset of target-indication pairs with no approved 2013 drugs for that target. Risk ratio and 95% confidence intervals.(PDF)Click here for additional data file.S13 TableReplication of Table 1N from Nelson et al. supplementary genetic association dataset and updated pipeline data with all MeSH terms assigned valid headings (All OMIM). Column New OMIM shows results using only OMIM entries originally mapped to supplementary concepts (and therefore not used in Table 1N). Risk ratio p(approved | genetic support)/p(approved | no genetic support) and bootstrap 95% confidence intervals.(PDF)Click here for additional data file.S14 Tablep-value or other cutoff used to filter results.Sources used to obtain and annotate GWAS variants. Filter gives the (PDF)Click here for additional data file.S15 TableComparison of counts of distinct genes, traits (MeSH), and SNPs reported by Nelson et al. and those from the current analysis. Overlap is the number of items in common.(PDF)Click here for additional data file.S16 TableComparison of counts of distinct genes, traits (MeSH), and SNPs reported by Nelson et al. and those from the current analysis, restricted to SNP associations expected to appear in both datasets . Overlap is the number of items in common.(PDF)Click here for additional data file.S17 TablePercent of LD SNP-Gene associations reported by Nelson et al. also found in the reanalysis by whether the association was an eQTL. Conditional on LD SNP presence in both analyses.(PDF)Click here for additional data file.S18 TablePercent of LD SNP-Gene associations found in reanalysis also reported by Nelson et al. subdivided by what evidence source(s) were used to link the LD SNP and gene. Conditional on LD SNP presence in both analyses.(PDF)Click here for additional data file.S19 TableReplication of Table 1N from updated GWAS Catalog and OMIM genetic association dataset and updated pipeline data (Full Data). Risk ratio p(approved | genetic support)/p(approved | no genetic support) and bootstrap 95% confidence intervals.(PDF)Click here for additional data file.S20 Tablep-value cutoff 10\u221212 .Replication of Nelson et al. Table 1 from updated GWAS Catalog and OMIM genetic association dataset and updated pipeline data, eQTL (PDF)Click here for additional data file.S21 TableEffect of OMIM genetic evidence on historical pipeline progression. Risk ratio p(approved | genetic support)/p(approved | no genetic support) and bootstrap 95% confidence intervals. Pharmaprojects drugs are subdivided by indication MeSH heading: Congenital indications are those indexed under Congenital, Hereditary, and Neonatal Diseases and Abnormalities and OMIM indications are those MeSH headings that are also mapped MeSH headings to an OMIM phenotype.(PDF)Click here for additional data file.S22 TableEstimated effect of GWAS genetic evidence from case-control studies on historical pipeline progression by effect size (odds ratio). Risk ratio p(approved | genetic support)/p(approved | no genetic support) and bootstrap 95% confidence intervals.(PDF)Click here for additional data file.S23 TableSimilarity between different pairs of traits computed using Resnik similarities, Lin similarities, normalized Resnik similarities, and average of normalized Resnik and Lin similarities using number of descendants to compute information content.(PDF)Click here for additional data file."} +{"text": "While the nuclear ribosomal internal transcribed spacer (ITS) region are recognized as the formal fungal taxonomic barcode, appraisal of different ITS sub-regions and the influence of DNA extraction methods have not been comprehensively undertaken using human respiratory specimens. (2) Methods: We performed ITS analysis of respiratory (sputum) samples by assessing (a) the effect of alternate DNA extraction techniques and (b) an evaluation of four different ITS primer pairs on the mycobiome profiles generated for mock fungal communities and their respective clinical (airway) specimens. (3) Results: Primer pairs varied in their resulting ITS mycobiome profiles, suggesting that particular pairs may be more relevant for analysis of respiratory samples compared to others. Assessment of DNA extraction methods highlighted lower final DNA concentrations achieved by mechanical disruption compared to enzymatic lysis. However, despite lower yields, DNA liberated by mechanical lysis more readily yielded ITS bands with highest success in combination with the Fseq and Rseq primers. (4) Conclusion: Choice of extraction method, primers used, and sequencing approach are all important considerations in sequencing the mycobiome and should be tailored to sample type. A standardization of approach to mycobiome studies using respiratory specimens will permit more reliable comparisons between studies and improve our understanding of the role of fungi in the human airway. Fungal disease affects more than 300 million people worldwide, accounting for an estimated 1.5 million deaths annually . While pHigh throughput sequencing (HTS) of the airway has uncovered novel associations between the pulmonary microbiome and chronic respiratory disease . To dateBlastomyces. They compared a range of primers targeting different ITS regions, representing an important benchmarking study for application on other respiratory disease states or specimen types such as sputum were used in this study. Clinical information on the recruited patients is provided in Twenty-one patient sputum samples [COPD were centrifuged at 13,000 rpm for 10 min and 500 \u03bcL sterile phosphate-buffered saline PBS was added to re-suspend resultant pellets. As for fungal cultures, the samples were homogenized through bead beating and the DNA was purified using the Roche High Pure PCR Template Preparation Kit . In parallel, 200 \u03bcL of selected sputum samples in RNAlater was used for DNA extraction and purification using the Quick-DNA\u2122 Miniprep Plus Kit according to the manufacturer\u2019s instructions. All purified DNA was quantified using the Qubit\u2122 fluorometer 2.0 double-stranded DNA (dsDNA) assay . The DNA quality was assessed using the NanoDrop\u2122 Spectrophotometer . Fungal cultures grown on SDA were scraped and mixed with 500 \u03bcL PBS. The samples were then transferred to sterile bead mill tubes (VWR) containing 1 mm sterile glass beads . Homogenization using a beads mill homogenizer was then performed followed by DNA purification using the Roche High Pure PCR Template Preparation Kit .Two types of mock community were prepared, referred to as fungal DNA (FD) and fungal Amp (FA) throughout. (i) FD: DNA from fungal isolates was extrhttps://bit.ly/30mczJZ) [All primers were synthesized with the respective forward and reverse overhang adaptors as described in Illumina ITS metagenomics demonstrated protocols (30mczJZ) . For eacAmplicons were purified with Agencourt AMPure XP beads . Library preparation was performed based on Illumina\u2019s ITS metagenomics demonstrated protocol . AgaroseTargeted amplicon sequences were analyzed using the ITS Metagenomics pipeline , using as a taxonomic reference database the v7.2 UNITE project described by K\u00f6ljalg et al. . ControlCollection of all clinical samples used in this study was approved by the respective Institutional Review Boards (IRB) as follows: NTU IRB-2016-01-031 (14/03/2016); NTU IRB-2017-03-013 (25/04/2017); CIRB 2017/2933 (31/01/2017); and CIRB 2016/2073 (22/03/2016)."} +{"text": "Fluid resuscitation is a cornerstone of the initial management of severely burned patients with the dual purpose of avoiding both under- and over-resuscitation \u20133. ThereThis international survey focuses on the current practices regarding hemodynamic management of severely burned adult patients \u2009>\u200920%, with mechanical ventilation) in the early phase after injury.The study was designed as an electronic survey addressed to intensive care unit (ICU) physicians. Experts of the European Society of Intensive Care Medicine (ESICM) Burn ICU working group were invited to review the original survey. The final questionnaire (32 questions) is provided in Additional\u00a0file\u00a0There were 173 total respondents to the questionnaire. The respondents were from 58 different countries (72% were high-income countries) with most in Europe (62%). The background of the respondents was mainly intensive care (61%) and anesthesiology (31%). Most of the respondents (61%) declared working in a mixed ICU, and 60% of the responders worked in centers with less than 50 adult burn patients admitted annually. Additional\u00a0file\u00a0The results of this international survey highlight the use of albumin (>\u200960%) and vasopressors (80%) during the early resuscitation phase. Heterogeneous results were reported regarding monitoring strategies, early vasopressors, and albumin use between burn centers and nonspecialized centers. Large clinical trials should be initiated in the near future to determine optimal strategies to treat burn-related shock.Additional file 1:Survey questions. (PDF 131 kb)Additional file 2:n number of respondents per group. The results are reported as numbers and percentages (%). The chi2 and Fischer tests were used as appropriate (p\u2009<\u20090.05). (PDF 155 kb)Comparison of participant responses between burn centers and nonspecialized centers. CO cardiac output,"} +{"text": "Oryza sativa L.) genome contains about 40 GLP family member proteins in nine chromosomes. Although some of the rice GLP (OsGLP) promoters have been studied through in silico analysis as well as experimentally, studies regarding the distribution pattern of the biotic and abiotic stress associated transcription factor binding sites (TFbs) in the promoter regions of OsGLP genes have not been attempted thoroughly. Several transcription factors (TFs) namely NAC, WRKY, bHLH, bZIP, MYB and AP2/ERF act as major TFs concerned with biotic as well as abiotic stress responses across various plant species. In the present study the in silico analysis was carried out using the 1.5 kilobases (kb) promoter regions from 40 different OsGLP genes for the presence of NAC, WRKY, bHLH, bZIP, MYB and AP2/ERF TFbs in it. Among various OsGLP gene promoters, OsGLP8-11 was found to contain highest number of tested TFbs in the promoter region whereas the promoter region of OsGLP5-1 depicted least amount of TFbs. Phylogenetic study of promoter regions of different OsGLP genes revealed four different clades. Our analyses could reveal the evolutionary significance of different OsGLP gene promoters. It can be presumed from the present findings as well as previous reports that OsGLP gene duplications and subsequent variations in the TFbs in OsGLP gene promoter regions might be the consequences of neofunctionalization of OsGLP genes and their promoters for biotic and abiotic stress tolerance in rice.Germin-like proteins (GLPs) are involved in biotic and abiotic stress tolerance in different plant species. Rice ( Germin-like proteins (GLP) belonging to cupin superfamily are evolutionary conserved plant glycoproteins . Germin Oryza sativa L.) is used as a major staple food for more than half of the world\u2019s population. Since the availability of genomic sequence information in public database, several studies have been conducted to characterize individual GLP family members from rice and other crops [OsGLP gene families as quantitative traits loci (QTL) associated with blast disease and all of those genes were present on chromosome number 8 [OsGLP1) depicted susceptibility of OsGLP1-down regulated plants to sheath blight and blast diseases of rice compared to untransformed control plants [OsGLP1 gene improved resistance to Fusarium solani infestation in the transgenic tobacco lines [GLP genes have been found to be involved in abiotic stress mechanisms also. RNAi-mediated down-regulation of OsGLP1 has documented improved salt tolerance of transgenic lines during seed germination and early seedling growth compared to untransformed plant [OsGLP genes upon exposure to drought, salt, cold and heavy metal stresses [Rice promoter showed wound inducible property as well as abiotic stress responsiveness [GUS fusion construct containing 1107 base pair (bp) promoter region of OsRGLP2, depicted expressional up-regulation of GUS fusion protein in response to two fungal pathogens namely F. solani and Alternaria solani [OsGLP1 and a putative germin A promoter region from five Pakistani rice varieties and it revealed that the TATA box binding protein could also recognize regions other than the (-30) TATA box element on the promoter sequences [GLP gene promoters have also been characterized in other crops. In an earlier study, a GLP gene (PcGER1) promoter was cloned from Pinus caribaea and the 1520 bp upstream promoter region was characterized in tobacco Bright Yellow 2 (BY\u20102) cells upon exposure to different phytohormones [Hordeum vulgare), eight different germin-like protein (GER4) gene promoters have been analyzed and it has been found that different promoters showed diverse response upon pathogen attack preferably due to the presence of W-box domain in the TATA-box proximal promoter region [GER4 gene, the promoter region rather than the coding DNA sequence (CDS) region have been diversified after duplication of the genomic region to subsequently acquire new function [In association with different siveness . Anotherequences . In addihormones . Maximumhormones . In barlr region . Anotherotein GER gene proPR genes) or signaling genes after binding with their promoter regions [The adaptive strategy of plants under biotic and abiotic stress conditions include expression as well as utilization of several transcription factors (TFs) which eventually regulate a number of pathogenesis related genes region of 40 OsGLP gene promoters have been analyzed for putative availability of the major biotic and abiotic stress-related transcription factor binding sites (TFbs). Additionally, the phylogenetic study of OsGLP gene promoters, their evolutionary significance and the expression of OsGLP genes under respective gene promoters have also been interpreted here.Although, a large number of GLPs are available in rice, their exact number and proper nomenclature have been found to be highly confusing , 20. A rl stages . Beside onducted , 21\u201322. onducted , 14. In OsGLP genes were collected from Rice Genome Annotation Project Database (https://rapdb.dna.affrc.go.jp/) and from National Center for Biotechnology Information database (https://www.ncbi.nlm.nih.gov/). All of the mRNA sequences for each OsGLP were retrieved from NCBI database. The coding DNA sequences (CDS) of each OsGLP were blasted at NCBI using Ref Seq Representative Genome database and their corresponding DNA sequence was identified from different rice chromosome. As the length of 5\u2032 UTR (untranslated region) of different OsGLP mRNA were found to differ in length [O. sativa Japonica Group cultivar Nipponbare.Protein sequences of n length , the upsOsGLP genes (i.e. 1.5 kb upstream regulatory region from CDS) was conducted through Neighbor-Joining method using Molecular Evolution Genetic Analysis 7 (MEGA 7) software [OsGLP gene promoters. All positions containing gaps and missing data were eliminated during evaluation. There were a total of 801 positions in the final dataset. Tajima\u2019s neutrality test was conducted using the above mentioned software for finding out nucleotide diversity. During selection of the model, nucleotide frequencies and rates of base substitutions for each nucleotide pair were considered in MEGA 7. Chromosomal organization of 40 OsGLP genes were performed with the help of Rice Database Oryzabase-Shigen (https://shigen.nig.ac.jp/rice/oryzabase/) using the Chromosome Map Tool (http://viewer.shigen.info/oryzavw/maptool/MapTool.do). Different OsGLP genes which were separated by a maximum of five genes were designated as duplicated genes in accordance to published protocol [OsGLP genes on positive or negative strand of the DNA, were counted for considering them as tandemly or inverse tandemly duplicated genes, respectively.Phylogenetic analysis of 40 different promoter regions of software . The perprotocol . The preOsGLP genes were analyzed using PlantPAN 2.0 software (http://plantpan2.itps.ncku.edu.tw/) for identification of transcription factor binding sites (TFbs) in OsGLP gene promoters. Multiple promoter analysis programme (http://plantpan2.itps.ncku.edu.tw/gene_group.php?#multipromoters) was used and occurrence of same TFbs in different promoter regions were identified. TFbs for different TFs involved in biotic and abiotic stresses namely NAC, WRKY, bHLH, bZIP, MYB and AP2/ERF were studied in 40 OsGLP gene promoters. A total of 1.5 kb upstream sequence was analyzed for each GLP member and it was divided into three regions namely proximal promoter region (500 bp upstream from the start codon), median promoter region (501 bp to 1000 bp upstream from the start codon) and distal promoter region (1001 bp to 1500 bp upstream from the start codon).Promoter regions of 40 OsGLP gene expression data was extracted by GENEVESTIGATOR from O. sativa database using Affymatrix Rice Genome Array platform (OS_AFFY_RICE-0) and \u2018Perturbations\u2019 tool was used to find out the differential gene expression under biotic and abiotic stresses. The fold change in gene expression was detected using filter 2.0 fold as benchmark. The fold changes in the expression of OsGLP genes under different biotic and abiotic stress conditions (http://www1.heatmapper.ca/expression/) using Red/Green colour scheme where \u201cRed\u201d colour shows down-regulation and \u201cGreen\u201d colour shows up-regulation of respective genes. The microarray dataset used for OsGLP gene expression study using GENEVESTIGATOR has been presented in nditions Files weOsGLP gene promoters of O. sativa retrieved from NCBI database have been depicted in OsGLP genes studied here and chromosomal locus of the respective OsGLP gene has been described and 3 OsGLP genes were identified in chromosome 9 (OsGLP genes from rice [OsGLP members have been characterized here. The CDS sequence of OsGLP5-2 belonging to locus LOC_Os05g19670 [Oryza sativa japonica Group OsGLP5-1 (accession number XM_015782936). Additionally, the protein sequence of OsGLP1-5 with locus number LOC_Os01g72300 [GLP (OsGLP3-9) belonging to the locus LOC_Os03g58990 mentioned in earlier report [OsGLP3-9 showed maximal homology with CDS sequence of OsGLP3-8 gene of Oryza sativa japonica group and no other mRNA from japonica group exhibited significant homology with that GLP. Hence, only 40 unique OsGLP gene promoters have been considered here. These large numbers of OsGLP gene promoters might have been generated following the destinies of duplicated OsGLP genes through neofunctionalization of the promoter region as revealed in barley GLP gene cluster [OsGLP genes exhibited tandem duplications while OsGLP8-13 and OsGLP8-14 exhibited inverse tandem duplications have been categorized on the basis of their involvement in ABA-independent and ABA-dependent pathways . In the OsGLP gene promoter regions NACbs were absent , which depicted significantly higher yield compared to the non-transgenic lines under drought stress condition [OsNAC9 under root-specific RCc3 promoter exhibited increased grain yield in exposure to drought stress [NAC in rice also improved tolerance to dehydration and high salinity, although with growth retardation and low reproductive yields [A total of 58 NAC TFbs were recognized in 40 different regions . The proe absent . Maximum signals . Expressstresses . Additiove stage . In anotondition . Similart stress . Accordie yields , 36.OsGLP gene promoter regions of target gene promoters to modulate transcription [In the present study, a total of 118 WRKY TFbs were detected covering all the regions . The prol region . Howeverectively . WRKY iscription , but acccription as well cription motifs icription . Moreovecription . Along wcription , 42. Thecription , 44.OsGLP promoters were detected with 107, 92 and 93 bHLHbs, respectively in both the strands. Among the OsGLP gene promoters the highest number of bHLHbs was found in OsGLP8-11 having 20 bs, while OsGLP5-1 was devoid of any bHLHbs for regulating gene expression [OsGLP gene promoters like OsGLP1-4, OsGLP3-5, OsGLP3-6 and OsGLP8-13 were found to possess \u201cCANNTG\u201d bs, most of them were found to have \u201cCANNTN\u201d sequence and it was recognized as probable bHLHbs. Previous findings confirmed the role of bHLH in inducing ABA-dependent signaling for management of cold stress [In the present study a total of 292 bHLH TFbs were recognized . The proy bHLHbs . Presentpression . Althougd stress . Moreoved stress . Additiod stress . In riced stress whereas;d stress .OsGLP gene promoters, the highest number of bZIP specific bs (21) was observed in OsGLP8-11 promoter, while the least bZIPbs (2 in each) were found in OsGLP1-3, OsGLP3-8 and OsGLP12-2 gene promoters gene signaling pathway and thus enable enhanced biotic stress tolerance in rice [It was found that a total of 232 bZIP TFbs were detected to be distributed throughout the studied promoter regions covering both the strands . Most ofromoters . Earlierignaling \u201352. Exteignaling . Neverthss genes . Beside in rice . Earlier in rice .OsGLP gene promoter regions among all the 40 OsGLP gene promoters . In the earlier reports it was validated that ERF families could bind with two cis-acting elements viz. GCC box and CRT elements, involved in ethylene responsiveness related to PR genes and expression of cold and dehydration responsive genes, respectively [OsGLP gene promoters. Several studies reported improved drought, salinity, water use efficiency, heat and cold response without compensating yield losses in transgenic rice plants through over-expression of AP2/ERF genes either in root-specific or in constitutive manner [PR genes after invasion by pathogen for enabling better resistance mechanism against pathogens [A total of 3238 bs were recognized in context to AP2/ERF TFs. Grippinromoters . Among tromoters . In the ectively . An inteectively and somee manner \u201365. Addie manner . Regardiathogens . It is wathogens \u201368.OsGLP gene promoters, largest number of TFbs was detected in OsGLP8-11 followed by OsGLP12-3, while least number of bs was identified in OsGLP5-1 followed by OsGLP1-2 promoter region [OsGLP8-11 might be highly responsive to drought stress. Further experimentation is needed with the high and low TFbs possessing OsGLP promoters to unravel their biological significance.In the present study on six TFbs availability among 40 OsGLP1-2 . AlthougsGLP8-11 , the higr region . Accordir region and it cOsGLP gene promoters were found to be divided into 4 different clades with bootstrap values of 0 to 100 from chromosome 8 and 12, while the cluster 2 had 3 sequences belonging to chromosome 8. Clade II was comprised of 6 sequences having two clusters where, cluster 1 contained 3 sequences from chromosome 3 and 4 while the cluster 2 was composed of 3 promoter sequences from 3 different chromosomes . The clade III was revealed as the smallest clade and it consisted of only 3 sequences; of which two (OsGLP3-1 and OsGLP3-3) were from chromosome 3 and one (OsGLP1-1) from chromosome 1. Clade IV was identified as the largest clade consisting of 16 promoter sequences of which 8 sequences were in cluster 1 and remaining 8 sequences were belonging to cluster 2. OsGLP promoters on the chromosome 12 showed maximum homology among themselves and OsGLP promoters on chromosome 8 also exhibited high level of similarity with each other were distantly related to the rest of the OsGLP promoters present in the same chromosome. In addition to the OsGLP promoters of chromosomes 8 and 12, promoters belonging to chromosome 2 were also grouped in a particular clade (clade IV). It was observed that on chromosomes 1, one OsGLP promoter (OsGLP1-1) was categorized under clade III while other three promoters were grouped in clade IV of chromosome 9, were grouped in clade IV while another promoter (OsGLP9-2) was grouped under clade II . Additionally, the nucleotide diversity among the tested sequences was 0.696. Most of the OsGLP promoters on chromosomes 8 and 12 shared the highest sequence similarity and this might be due to the duplication of the genomic region including the presence of TFbs. In corroboration to our findings, OsGLP promoters analyzed using 1.0 kb promoter region also depicted the relatedness of the promoters on chromosomes 8 and 12 [viz. OsGLP8-11, -13 and -14 were distantly related, earlier study [OsGLP8-11, -12 and -13 promoters were phylogenetically distinct from the rest of the OsGLP promoters on chromosome 8. This difference with earlier study might be due to the consideration of 1.5 kb promoter region in our study. TFbs analysis revealed that the OsGLP8-14 promoters was devoid of any NACbs and only one WRKYbs was available in the distal promoter region making it unique from rest of the OsGLP promoters of chromosome 8 and different GER4 promoters exhibited differential expression in response to biotic stress due to the available changes in their cis-regulatory region [In accordance with the previous report , in the lication so, theslication . In a siers also . Anothery region .OsGLP gene expression and OsGLP gene promoter classification. Gene expression of OsGLP8-3 and OsGLP8-4 depicted retarded gene expression under several biotic stresses while the expression study by microarray demonstrated up-regulated gene expression for OsGLP8-7 and OsGLP8-10 in response to brown plant hopper (Nilaparvata lugens), Agrobacterium tumefaciens and fungal infestations were found to possess larger number of TFbs and some promoters had fewer, further study is needed to validate these OsGLP promoters for subsequent utilization in plant genetic engineering as stress inducible promoter. Gene expression data revealed that certain promoters belonging to same clade had similar pattern of gene expression. It can be concluded that OsGLP gene duplication and subsequent variation in TFbs resulted neofunctionalization of gene and promoter regions to cope up with various biotic and abiotic stresses during the course of evolution.In the present study among 40 S1 Table(DOC)Click here for additional data file.S1 File(DOC)Click here for additional data file.S2 File(XLS)Click here for additional data file.S3 File(XLS)Click here for additional data file."} +{"text": "Introduction: Blue light cystoscopy (BLC) using hexaminolevulinate (Cysview\u00ae) improves the detection of nonmuscle invasive bladder cancer (NMIBC).1\u20133 BLC results in lower recurrence rate and a better recurrence-free survival, as well as a progression benefit.4 However, false-positive (FP) fluorescence can occur for various reasons and can vary among different series. Studies have shown that FP rates are not significantly different from white light (WL) cystoscopy. We evaluated different scenarios producing FP in BLC.Methods: Under institutional review board approval, we prospectively enrolled consecutive patients undergoing transurethral resection of bladder lesions into a BLC registry between April 2014 and December 2016. Several cases are highlighted in the video demonstrating cystoscopic view under WL and blue light in specific circumstances increasing the chance of detecting an FP lesion.Results: BLC with Cysview is demonstrated in several challenging cases for the detection of NMIBC. Possible FP scenarios include tangential views of the bladder neck or side walls (1) trigone, trabeculations, or diverticula; (2) in setting of inflammation like cystitis; (3) postintravesical therapy, that is, <6 weeks interval from prior bacillus Calmette-Gu\u00e9rin (BCG); (4) prior resection within 6 weeks; (5) bright tiny spots; and (6) site of ureterectomy/bladder cuff resection, early fading lesions (after irrigation). Unnecessary biopsy of these lesions can be avoided through simple techniques such as changing the angle of the cystoscopic view, several rounds of irrigation, and avoiding BLC too early after BCG instillation or prior resection.Conclusions: Use of BLC with Cysview can help with the detection of NMIBC as well as carcinoma in situ in patients undergoing transurethral resection of bladder tumor for bladder cancer. The reported FP rates of BLC will decrease with experience and recognition of the mentioned scenarios.Prior presentation: None.No competing financial interests exist.Runtime of video: 7\u2009mins 16\u2009secs"} +{"text": "Solanum lycopersicum L. (Tomato) is an economically important crop plant and model system for various studies with massive genomic data. The comprehensive identification and characterization of P450 genes was lacking. Probing tomato genome for P450 identification would provide valuable information about the functions and evolution of the P450 gene family.Cytochrome P450 (P450) is a functionally diverse and multifamily class of enzymes which catalyses vast variety of biochemical reactions. P450 genes play regulatory role in growth, development and secondary metabolite biosynthesis. Solanum lycopersicum P450 (SlP450) protein sequences, they were classified into two major clades and nine clans further divided into 42 families. RT-qPCR analysis of selected six candidate genes were corroborated with digital expression profile. Out of 233 SlP450 genes, 73 showed expression evidence in 19 tissues of tomato. Out of 22 intron gain/loss positions, two positions were conserved in tomato P450 genes supporting intron late theory of intron evolution in SlP450 families. The comparison between tomato and other related plant P450s families showed that CYP728, CYP733, CYP80, CYP92, CYP736 and CYP749 families have been evolved in tomato and few higher plants whereas lost from Arabidopsis. The global promoter analysis of SlP450 against all the protein coding genes, coupled with expression data, revealed statistical overrepresentation of few promoter motifs in SlP450 genes which were highly expressed in specific tissue of tomato. Hence, these identified promoter motifs can be pursued further as tissue specific promoter that are driving expression of respective SlP450.In the present study, we have identified 233 P450 genes from tomato genome along with conserved motifs. Through the phylogenetic analysis of SlP450 gene family with other Solanaceae members which are also economically important and attempt to classify functionally important SlP450 genes into groups and families. This report would enable researchers working on Tomato P450 to select appropriate candidate genes from huge repertoire of P450 genes depending on their phylogenetic class, tissue specific expression and promoter prevalence.The phylogenetic analysis and expression profiles of tomato P450 gene family offers essential genomic resource for their functional characterization. This study allows comparison of The online version of this article (10.1186/s12864-019-5483-x) contains supplementary material, which is available to authorized users. Arabidopsis. Most of the P450 studied in plants are localized in the endoplasmic reticulum, chloroplast or mitochondria and other secretary pathways but more than mulberry (176) . Hence, SlCYP80E6 is a potential candidate to study the floral development. The expression data suggests that SlCYP84A2 gene was up-regulated in root and has root specific overrepresented AGL promoter motif. In Arabidopsis, CYP84A1 gene is involved in the lignin biosynthesis. The functional analysis of this gene affects the lignification and vascular development [SlCYP84A2 gene might be involved in vascular development of the root.Cytochrome P450 genes are involved in catalysis of variety of reactions which include growth, development and secondary metabolite biosynthetic pathways. In present study we identified 233 P450 genes from tomato which are comparable with genes identified in ry 176) . All ide6 . All ispecific . The CYPr plants , 52, 53.sistance , 58.\u00a0Theue types . The CYPue types . The SlCArabidopsis P450 revealed similar clustering that indicates conserved nature of P450 multigene family across the various plant species [SlCYP51G1 showed 82% identity with AtCYP51G1 and it is involved in sterol metabolism [SlCYP51G1 is constitutively expressed in all selected tissues of tomato and has sterol demethylase activity required for the maintenance of membrane integrity [Arabidopsis CYP71 family. The expression data of SlCYP71AX and SlCYP77A20 from to clan71 showed that these two genes were up-regulated in green and mature green fruit of tomato which is in accordance with the transcriptome data. These two genes would be good candidates for the study of secondary metabolite synthesis in tomato fruit\u00a0[Phylogenetic tree topology of tomato and species . The sin species . The CYPtabolism . RT-qPCRntegrity . The CYPntegrity , 61. Tomto fruit\u00a0, 63.SlCYP74C3 gene in mature green tomato fruits and hence it could be a potential candidate gene to study oxylipin biosynthesis in tomato fruit. The SlCYP90A5 gene was up-regulated in tomato flower and showed less expression in leaf which correlate with transcriptome data. The AtCYP90A1 is involved in brassinosteroid metabolism and shows less expression in expanding leaf [SlCYP90A5 has similar expression profile. CBF/DREB1 transcription factor plays role in cold response [SlCYP72A184, SlCYP85A1 and SlCYP96A48 genes. These genes can be candidate for cold stress tolerance in tomato. Intron map along with their phases and gain/loss events plays a crucial role in understanding the evolution of gene families within phylogenetic group. Conserved introns are ancient elements and present with similar intron phase [Arabidopsis, two conserved introns were absent from non-A type P450 gene families whereas they appeared in A-type P450 gene families [SlP450 genes during the course of evolution. Intron gain was observed in the A-type of SlP450 genes which was absent in the ancestral (Non-A) gene families. Hence, this data support the intron late view of intron evolution [The CYP74 family is an atypical plant P450 family and thought to be involved in catalysis of already oxygenated polyunsaturated C18 fatty acid hydroperoxide into other oxylipins [ing leaf , 5 whereresponse and is oon phase . Intron on phase . Intronson phase . Both cofamilies . It is ovolution , 31.SlP450 promoter motifs are driving tissue specific expression. Thus present study may enable researchers to select appropriate candidate gene from huge repertoire of SlP450 for detailed functional characterization.The expression evidence to the genes profoundly depends on developmental stages, age of the plant, environmental conditions, extent of expression, tissue specificity and biotic or abiotic stress. In the present study, only 31.33% P450 genes showed evidence of expression which could be compared with rice (49.81%) and soybArabidopsis with variable number of P450 genes in each clan. Phylogenetic tree analysis provided the information about the functional evolution of P450 gene family in tomato. In intron map, gain and loss of conserved introns reveals P450 gene family evolution in tomato plant. Digital and experimental expression profile suggests tissues specific highly expressed P450 genes that could be potential candidates for further study. The promoter motifs driving the higher expression of P450 in analysed tissues types can be further evaluated using functional genomics for traits of economic importance. Thus, this study provides solid foundation for functional characterization of candidate genes with their biological significance.The Tomato genome has a greater number of P450 clans as compared to Additional file 1:SlP450 summary file\u00a0233 SlP450 sequences are provided with the universal names, Sol genomics id and phylogenetic groups. The NCBI accession numbers of Arabidopsis, Poplar and Potato P450 protein sequences used in phylogeny are listed. The sheet 2 represents percent identity matrix of all the SlP450 proteins. (XLSX 197 kb)Table of Additional file 2:Table of primer sequences used in experimentation. Primer sequences of genes used RT-qPCR analysis. (XLSX 8 kb)Additional file 3:SlP450 phylogenetic tree inferred using Maximum likelihood method. Phylogenetic tree is constructed by applying maximum likelihood method with 1000 ultrafast bootstrap replicates using LG\u2009+\u2009F\u2009+\u2009I\u2009+\u2009G4 as best-fit substitution model. (TIF 13827 kb)Additional file 4:Intron analysis summary. Data for Intron map constructions and distribution. (XLSX 34 kb)Additional file 5:Promoter analysis Table. Promoter analysis of all the protein coding genes in tomato with promoter motifs count. (XLSX 81994 kb)Additional file 6:Tissue specific expression of P450 with FPKM count. Highly expressed tomato P450 genes from heat map with their FPKM count. (XLSX 11 kb)Additional file 7:Table of P450 orthologue gene count in related plants. Comparison of tomato P450 gene along with other related plants P450 orthologue gene count. (DOCX 20 kb)"} +{"text": "Scientific Reports 10.1038/s41598-017-13639-y, published online 18 October 2017Correction to: This Article contained an error in the Acknowledgements section.\u201cThis study was funded by Shanghai Municipal Commission of Health and Family Planning grant number 140613163312706.\u201dnow reads:\u201cThis study was funded by Shanghai Municipal Commission of Health and Family Planning grant number 201440040.\u201dIn addition, the email addresses for Chengde Yang and Sheng Chen were incorrectly listed as 13917556052@139.com and yangchengde@hotmail.com respectively."} +{"text": "Rbm24 causes embryonic lethality limiting the functional analyses -factors. In addition, several regulators of alternative splicing like muscleblind like splicing regulator 1 (MBNL1) was the only known splicing factor of TTN in 190 cardiomyopathy index patients by Sanger sequencing. To our surprise, we did not find any pathogenic RBM24 mutation. Although we cannot exclude that in further specific cases pathogenic RBM24 mutations might be found, our data suggest that RBM24 mutations are rare in human cardiomyopathy patients. Thus, the data of our cohort suggests an allele frequency in non-ischemic cardiomyopathy patients below <0.5%. These findings are supported by the fact that only the homozygous Rbm24 deficient mice developed DCM in the study by Liu and colleagues (As RBM24 was already established as a major regulator of muscle-specific alternative splicing (Yang et al., lleagues .RBM24 can be established as a rare cardiomyopathy associated gene. However, with relevance for clinical cardiovascular genetics this gene deserves further attention in genetic screenings.In summary, it remains an open question if"} +{"text": "Vesicoureteral reflux (VUR) is a complex, heritable disorder. Genome-wide linkage analyses of families affected by VUR have revealed multiple genomic loci linked to VUR. These loci normally harbor a number of genes whose biologically functional variant is yet to be identified. DNA copy number variations (CNVs) have not been extensively studied at high resolution in VUR patients. In this study, we performed array comparative genomic hybridization (aCGH) on a cohort of patients with a history of both VUR and urinary tract infection (UTI) with the objective of identifying genetic variations responsible for VUR and/or UTI susceptibility. UTI/VUR-associated CNVs were identified by aCGH results from the 192 Randomized Intervention for Children With Vesicoureteral Reflux (RIVUR) patients compared to 683 controls. Rare, large CNVs that are likely pathogenic and lead to VUR development were identified using stringent analysis criteria. Because UTI is a common affliction with multiple risk factors, we utilized standard analysis to identify potential disease-modifying CNVs that can contribute to UTI risk. Gene ontology analysis identified that CNVs in innate immunity and development genes were enriched in RIVUR patients. CNVs affecting innate immune genes may contribute to UTI susceptibility in VUR patients and may provide the first step in assisting clinical medicine in determining adverse outcome risk in children with VUR. Primary vesiocoureteral reflux (VUR), characterized by retrograde flow of urine from the bladder to the ureter, affects 1\u20132% of children. MultiplThe Randomized Intervention for Children with Vesicoureteral Reflux (RIVUR) trial randomized children with a history of UTI and VUR to daily antibiotic prophylaxis (AP) or placebo and followed them for two years to monitor for UTI recurrence risk. Daily AVUR is a heritable condition. One third of siblings of children with VUR will also have the condition . AdditioCopy number variations (CNV) represent structural genomic variations comprising chromosomal segments that deviate from classic Mendelian diploid gene numbers . HundredBecause VUR severity and UTI burden are variable, we hypothesized that CNVs in pathways critical to innate immunity and renal development will segregate with distinct clinical outcomes and phenotypes in VUR patients. We used high-resolution array comparative genomic hybridization (aCGH)-based genetic analysis to determine the CNV profile of children from The Randomized Intervention for Children with Vesicoureteral Reflux (RIVUR) trial with VUR and UTI and compared to race/gender-matched controls.. To identify VUR and UTI associated CNVs, we assessed the CNV frequencies in genomic DNA using standard and stringent CNV calling criteria and results filtering . Because VUR is a relatively common disorder (1\u20132% of population) and UTIs are even more common, we utilized two different analysis methods in this study. Standard methodology resulted in a comprehensive candidate list, but difficult to interpret clinical relevance. We further utilized stringent analysis to look for strongly significant \u201ctrue positive\u201d candidates at the expense of eliminating some candidates that may be clinically relevant.We performed high-resolution genome-wide aCGH arrays on 192 RIVUR patients and compared the results to 683 gender/race/ethnicity-matched controls . The sizes of these rare CNVs detected by aCGH have a median CNV size of 7,770 bp . Among all the CNVs, 47.1% represent copy number loss . The chromosomal locations and size distribution of the rare CNVs were analyzed and shown in Fig 2A and 2B. Additionally, among all the gene-affecting CNVs, 61.8% were found to intersect genomic regions that encode protein sequences. The remaining non-coding region encompassed genetic components including antisense (9.9%), long intergenic non-coding RNA (lincRNA) (8.8%), pseudogene (7.7%), non-coding RNA (5.5%), etc .Our stringent analysis resulted in identification of 85 significantly altered chromosomal regions with differential CNV frequencies passed our quality control filtering under the stringent analysis criteria. S3 Fig shows the top 20 most significantly altered rare CNVs and their affected genes in RIVUR patients versus controls. Among all the differentially altered CNVs, the chromosomal locus\u2013chr12:11,215,990\u201311,217,836, is the top ranked CNV region harboring a 1.84-kb DNA segment of copy number gain. We detected this CNV in 33 (17.1%) of the RIVUR patients as compared with 0 controls, showing a significant enrichment (adjusted P = 4.84 x 10\u221221). This region harbors Proline Rich Protein HaeIII Subfamily 1 (PRH1) and two readthrough genes, PRH1-PRR4 and PRH1-TAS2R14.Within the significantly altered CNVs , 85 candidate CNVs affecting 141 genes . The GO analysis revealed multiple enrichments for developmental pathways. Of note, \u201cregulation of Wnt signaling pathway\u201d (P = 0.0008) and \u201cdevelopmental growth involved in morphogenesis\u201d (P = 0.018) are represented in this list. These pathways contain statistically significantly altered genes that have known roles in kidney development and/or known causes of VUR. The complete GO pathway analysis is presented in S3 Table. Additionally, we filtered our results against RefSeq for terms consistent with kidney and lower urinary tract development (see complete FGFR3) and Tenascin XB (TNXB) (Table 1and S2 Table) [Table 2) [To gain insight into the potential biological consequence of rare CNVs, we performed GO pathway analysis of the candidate CNVs 2 Table) \u201314. FinaTable 2) , 15\u201323.. Thirty-four percent of these common CNVs were copy number loss events. Common CNVs were smaller and less likely to be losses compared to rare CNVs . S5 Fig shows the top 20 most significantly altered comme and rare CNVs and their affected genes in RIVUR patients versus controls.Unlike classic genetic studies, we also performed an analysis on \u201ccommon\u201d CNVs. We justify this approach as UTIs are a common condition and multifactorial, thus multiple genes likely contribute to innate defenses of the kidney and urinary tract. Our standard analysis resulted in identification of multiple significantly altered chromosomal regions with differential CNV frequencies We have combined several similar pathways from S6 Table to highlight statistically significant unique processes in We identified pathways critical in collecting duct development, innate immune response and receptor signaling. Furthermore, CNVs within enriched pathways included genes that encode antimicrobial peptides such as and bacterial agglutins such as Deleted in Malignant Brain Tumor 1 (DMBT1).GO pathway analysis of common CNVs significantly associated with VUR/UTI based on adjusted P values are presented in S7 Table)(Comprehensive Methods for terms). We identified CNVs with potential innate immune functions involved genes that involve cytokines or chemokines such as Interleukin 11 (IL11), pattern recognition receptors such as Toll-Like Receptor 9 (TLR9), and antimicrobial peptides such as azurocidin 1 (AZU1) [.By searching RefSeq, we queried gene function of statistically significant CNV-associated genes for known roles in innate immunity (1 (AZU1) , 25 has previously been associated with VUR [TNXB single nucleotide polymorphism was postulated to be a gain-of-function event. Interestingly, in RIVUR, CNVs in TNXB predicted as coding sequence variants were gain events, which correlates with the previously known genetic events.Prior research has focused on the developmental cause of VUR, and a number of murine models of VUR exist. These models indicate that the induction site of the ureteric bud during early kidney development gives rise to an abnormally tunneled ureter in the bladder and a subsequent faulty ureterovesical junction that leads to VUR . Becausewith VUR . TNXB siTNXB1, we identified additional CNVs affecting genes that have been implicated in lower urinary tract development. SRY-Box11(SOX11) has been reported to play a key regulatory role in renal development and its disruption has been implicated in causing congenital anomalies of the kidney and urinary tract [FGF) Receptor 3 is expressed during normal kidney development [NOTCH1 [DACT2) was identified in our studied and has a known role in collecting duct development [In addition to ry tract . Fibroblelopment , 35. NOT [NOTCH1 . Disheve [NOTCH1 . Additioelopment . Finallyin utero VUR are unknown, we essentially cannot find a \u201cpure\u201d comparison cohort that we are certain does not have a history of VUR even if a voiding cystourethrogram was performed for reasons other than urinary tract abnormalities or UTI.This study provides new insights into VUR/UTI pathophysiology, however we do acknowledge some limitations. While our aCGH provides very high-resolution data, we used 2 different aCGH platforms in a portion of the RIVUR patients due to commercial availability. We have run a subset of the same samples on both platforms and confirmed that the Agilent array does not detect additional CNVs found in the Nimblegen array. We did, however, demonstrate that less CNVs are detected, which we postulate is secondary to the lower resolution of the array. Additionally, the SFARI control cohort used a different reference genome. To account for these analysis factors, we filtered results for both aCGH array type and reference genome. We have also used SMASH sequencing to establish if any reference genome CNVs exist and filtered our results accordingly. Because the immortalization process to create our DNA source can result in differential genomic structural variations, we have filtered our results to exclude regions implicated . FinallyDevelopment of a genetic panel to identify patients at risk for sequalae such as recurrent UTI and subsequent renal scarring would help \u201clow risk\u201d children avoid unneeded antibiotics and radiation exposure as well as select \u201chigh risk\u201d patients for more aggressive treatment. Because the initial step of using genetic profiles is to improve care of children with VUR, we have identified several novel findings relevant to VUR/UTI pathophysiology including: 1) VUR patients with a UTI history are more likely to have CNVs involving innate immune genes compared to controls, 2) VUR patients with a UTI history are more likely to have CNVs that involve ureteric bud/collecting duct development pathways than controls, and 3) aCGH identified overlap between VUR loci identified in this study and prior linkage studies. Because the clinical course of children with vesicoureteral reflux is what is critical in clinical practice, determining those at risk for UTI is essential in managing children who are diagnosed with vesicoureteral reflux. Results from our study will serve as the foundation to inform medical decision-making and the first step in personalized medicine for this patient population.S1 File). The workflow of the overall experimental design is outlined in S1 Fig. Approval on human subjects was obtained by Nationwide Children\u2019s Hospital Institutional Review Board (IRB) protocols IRB07-00383 and IRB10-00319 and the University of Tennessee Health Science Center IRB protocol 14-03325-XP. All clinical investigations were conducted according to the principles expressed in the Declaration of Helsinki.For complete methods, please refer to the supporting information file (S1 File). [RIVUR cohort (cases): Randomized Intervention for Children with Vesicoureteral Reflux (RIVUR) Study : For complete study design and outcome data, please refer to previously published materials and the supporting information file (https://sfari.org). We collected results from patients from the Simons-Simplex Collection whose DNA was analyzed with the Nimblegen HD2 2.1 million probe microarray platform (3). The complex trait of idiopathic autism is not related to any developmental or innate immunity phenotypes of VUR.SFARI cohort (controls): To use samples of unrelated individuals, we selected unaffected mothers from families of Simons Foundation Autism Research Initiative (SFARI) cohort for case-control comparison . The SFALocal control cohort (controls): A total of 19, ethnicity/race-, sex- matched healthy controls with no prior history of VUR or UTI served as the control group in this study. These 19 control subjects\u2019 genomes were interrogated on the Nimblegen HD2 platform with the same reference genome as the RIVUR subjects. Additionally, these 19 ethnicity/race-, sex- matched local healthy controls were also used for filtering purposes. Specifically, the SFARI cohort and our local controls were interrogated on the same Nimblegen HD2 platform but with different reference genomes. In order to control for potential reference-specific CNVs, we compared the CNV frequency difference between the 19 normal controls (which had the same reference as the RIVUR samples) and the SFARI cohort. Whenever a 10% threshold of difference is identified, the involved CNV was labeled as dubious positive and excluded from all the downstream analysis as it was most likely attributed to a reference genome CNV.For case-control comparison, the case group consists of 192 non-Hispanic, Caucasian females from the RIVUR cohort. The control group is composed of 19 healthy controls as well as 664 unrelated sex and race/ethnicity matched samples from SFARI cohort.S1 File).We identified the copy number variants (CNVs) of reference DNA samples by SMASH . A detaiS1 File) [For our DNA quantification as well as high-resolution aCGH methodology, please refer to our previous work and the supporting information file (S1 File) .S1 File).The aCGH data was processed using Nexus 8 Copy Number software . For specific copy calling parameters, please refer to the supporting information file .We peformed filtering to standardize for array platform (Nimblegen vs. Agilent) and reference DNA. In order to correct for the possible genomic changes in lymphoblastic cell line, we specifically filtered out genomic regions that harbor putative LCL-sepcific changes before further analysis. (lymphoblastic cell line derived vs. primThe CNVs were classified into common or rare (< 1% in controls) CNVs based on their frequency in control group. The gene ontology enrichment pathway analysis were performed on the selected candidate genes. The information of clinical significance and most severe effect for each candidate CNV were retrieved from the ClinVar and the Ensemble Variant Effect Predictor databases, respectively.The rare candidate CNV-affected genes were mapped to the previously identified VUR susceptibility loci , 15\u201323.S1 Fig(EPS)Click here for additional data file.S2 Fig(A) shows the cumulative distribution of CNVs by size. (B) reflects the distribution of copy number (CN) gains versus CN losses of rare candidate CNVs identified using stringent analysis criteria.(EPS)Click here for additional data file.S3 FigP) are plotted. * indicates the adjusted\u2014log10(P) values. The dashed black line shows where the significant threshold (P = 0.05) lies.The frequency differences (bar) and the level of significance transformed as\u2014log10((EPS)Click here for additional data file.S4 Fig(A) A higher total percentage of common CNVs are smaller compared to rare CNVs using standard analysis criteria.(B) A majority of common CNVs are gains, while the majority of rare CNVs are loss events.(EPS)Click here for additional data file.S5 FigThe top 20 common (A) and rare (B) disease-associated CNVs and their affected genes identified using standard criteria. The frequency differences (bar) and the level of signifi- cance transformed as\u2014log10(P) are plotted. * indicates the adjusted\u2014log10(P) values. The dashed black line shows where the significant threshold (P = 0.05) lies.(EPS)Click here for additional data file.S6 FigDMBT1 or WWOX gene locus, respectively. Panel C shows the entire genome with probes, a moving average line and colored chromosomes linked end to end.Panel A and B show that a copy number loss or gain is detected within the (EPS)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(DOCX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.S6 Table(XLSX)Click here for additional data file.S7 Table(DOCX)Click here for additional data file.S1 File(DOC)Click here for additional data file."} +{"text": "CYP) 2D6 gene, or to identify the promoter (TA)7 TAA repeat polymorphism UDP glucuronosyltransferase (UGT) 1A1*28. Here we developed and validated PGxSeq, a targeted exome panel for pharmacogenes pertinent to drug disposition and/or response.Targeted next-generation sequencing (NGS) enables rapid identification of common and rare genetic variation. The detection of variants contributing to therapeutic drug response or adverse effects is essential for implementation of individualized pharmacotherapy. Successful application of short-read based NGS to pharmacogenes with high sequence homology, nearby pseudogenes and complex structure has been previously shown despite anticipated technical challenges. However, little is known regarding the utility of such panels to detect copy number variation (CNV) in the highly polymorphic cytochrome P450 . Identified SNVs were assessed in terms of population allele frequency and predicted functional effects through in silico algorithms.A panel of capture probes was generated to assess 422\u2009kb of total coding region in 100 pharmacogenes. NGS was carried out in 235 subjects, and sequencing performance and accuracy of variant discovery validated in clinically relevant pharmacogenes. CYP2D6 CNV and UGT1A1*28. Allele frequency of rare or novel variants and predicted function in 235 subjects mirrored findings from large genomic datasets. A large proportion of patients were identified as homozygous carriers of at least one variant necessitating altered pharmacotherapy.Adequate performance of the PGxSeq panel was demonstrated with a depth-of-coverage (DOC)\u2009\u2265\u200920\u00d7 for at least 94% of the target sequence. We showed accurate detection of 39 clinically relevant gene variants compared to standard genotyping techniques (99.9% concordance), including PGxSeq can serve as a comprehensive, rapid, and reliable approach for the detection of common and novel SNVs in pharmacogenes benefiting the emerging field of precision medicine.The online version of this article (10.1186/s12920-019-0527-2) contains supplementary material, which is available to authorized users. Rapid identification of genetic variation contributing to therapeutic drug response or adverse effects is essential for implementation of individualized pharmacotherapy . Many geCYP) 2C9 and warfarin dose requirement content within sequencing reads (n\u2009=\u2009246). Histogram of the average percent GC content across total reads (A). Relationship between subjects average GC content and coverage (B). Figure S4. Mean (\u00b1SD) depth of coverage (DOC) across the targeted sequence for CES1 and CBR1 showing the inaccessible target regions. Figure S5. Study minor allele frequencies (MAF) in relation to the reported MAF in 1000 Genomes Project (1000G) and Exome Aggregation Consortium (ExAC) datasets. Figure S6. In silico functional prediction scores for genetic variants identified among 235 subjects. Rare or novel variations had a greater proportion of possibly deleterious prediction scores for all three algorithms . Figure S7. Zygosity of the potentially deleterious variants per subject (n\u2009=\u2009235), showing there were more heterozygous compared to homozygous variants per subject. Figure S8. Single nucleotide variants (SNV) per subject (n\u2009=\u2009235) found in cytochrome P450 (CYP) enzymes that are potentially deleterious variants separated by zygosity. Figure S9. Number of Pharmacogenomics Knowledge Base (PharmGKB) \u201cLevel 1A/1B\u201d variants found in 235 subjects separated by zygosity. Figure S10. Histogram of the GSTM1 and GSTT1 gene coverage as a fraction total subject coverage in 235 subjects. 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A large body of evidence indicates that, within the nucleus, chromatinized DNA is spatially organized at multiple length scales. The higher-order organization of chromatin is crucial for proper execution of multiple genome functions, including DNA replication and transcription. Here, we review our current knowledge on the spatial organization of chromatin in the nucleus of mammalian cells, focusing in particular on how chromatin is radially arranged with respect to the nuclear lamina. We then discuss the possible mechanisms by which the radial organization of chromatin in the cell nucleus is established. Lastly, we propose a unifying model of nuclear spatial organization, and suggest novel approaches to test it. Accordis (CTCF) . TAD bors (CTCF) , whereass (CTCF) . B compas (CTCF) . Dependis (CTCF) as well s (CTCF) , suggests (CTCF) , Dip-C (s (CTCF) , and sins (CTCF) , togethes (CTCF) , have main situ hybridization (FISH), which allows measuring the distance of chromosomes or individual genomic loci from each other or from defined nuclear structures, in single cells. More recently, a new method named TSA-seq was developed to infer the relative distance from nuclear speckles of thousands of genomic loci simultaneously, based on next-generation sequencing \u2014the three major constituents of the nuclear lamina\u2014led to condensation of heterochromatin in the nuclear interior . The res nucleus .In addition to specific nuclear proteins, histone modifications, and transcription, other factors have been suggested to contribute to shaping chromatin radiality. Genome-intrinsic features, such as chromosome size, guanine-cytosine (GC)-content, gene density, as well as the type and density of repetitive elements along the genome have long been associated with the radial arrangement of chromatin in the nucleus, in different human and mouse cell types . Indeed,e.g., chromatin-bound factors and/or histone modifications) or by long non-coding RNAs (lncRNAs). Indeed, several lncRNAs have been implicated in reorganizing genome architecture and initiating the formation of nuclear compartments , the Swedish Cancer Research Foundation (CAN 2015/585), the Ragnar S\u00f6derberg Foundation (Fellows in Medicine 2016), and the Strategic Research Programme in Cancer (StratCan) at Karolinska Institutet to NC; and by grants from the Science for Life Laboratory, the Karolinska Institutet KID Funding Program, the Swedish Research Council (621-2014-5503), the Human Frontier Science Program (CDA-00033/2016-C), the Ragnar S\u00f6derberg Foundation (Fellows in Medicine 2016), and the European Research Council under the European Union\u2019s Horizon 2020 research and innovation programme (StG-2016_GENOMIS_715727) to MB.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "A new viral illness called coronavirus disease 2019 (COVID-19) is currently spreading throughout the world at an alarming rate . As famiThe assumption that disease and its prevention is a family affair is manifested in the full spectrum and scale of the current coronavirus pandemic. Measures that have been taken in many countries to control the spread of the coronavirus are having a disruptive effect on relationships in general and family relationships specifically. Families are reporting loss of community and freedom of movement in response to quarantine/lock down measures. Other tangible losses include income, access to resources, and planned activities or celebrations. Compelling and heartbreaking stories of the challenges and suffering that families are experiencing engulf us. Individuals and families who are most vulnerable are particularly at risk.Residents in long-term care facilities miss their partners and children who are no longer allowed to visit because of the COVID-19 policies to contain the spread of the coronavirus. People with intellectual disabilities who live in institutions are upset because their father, mother, brothers, or sisters are no longer allowed to visit, and they often cannot understand why.Distressing stories abound of patients who have to deal with the news of their COVID-19 diagnosis all by themselves without a family member present and those patients who are admitted to an intensive care unit (ICU) who have to say goodbye to their family in the emergency department not knowing whether they will see each other again. A nurse working at the front line of triage reported, \u201cHis family was not allowed to come to the hospital, because they may also be infected. He was alone and couldn\u2019t say goodbye.\u201dMany of these patients, clients, and residents are members of families who miss their loved ones and who are worried. Mothers, fathers, and other family members of children receiving psychiatric care report being unable visit their child for an extended period of time and are afraid their child will become ill from the coronavirus. Families of very seriously ill and dying patients are not allowed to visit their loved one and may not be able to say a final goodbye.The lockdown/quarantine measures instituted in many countries have also invited vulnerability and risk within families. Schools are being closed which leads to distress in many families not accustomed to being so closely confined for a long time period. Moreover, as a result of the COVID-19 crisis, much, if not all, of the support given to families who provide long-term care for an ill parent, partner, or child is lost. Families with a child who requires specialized care and guidance now have to care for their child 24 hr a day without the outside guidance provided by medical nursery, daycare, or special education services. Families who care for a father, mother, or partner with dementia or other serious illness now have to manage without day care or daytime activity. School closures have created a family environment where children are rarely allowed to leave the house and are confronted by the vulnerabilities of a family member\u2019s addiction, aggression, and violence. Children of divorced co-parents are suddenly being refused alternating parental care because one of the parents now works from home and cannot provide child care. All of these families and their interrelationships are often under great pressure as a result of the stresses created by coronavirus pandemic.Health care professionals, including nurses and doctors, are also going through a very intensive and perhaps traumatic time. As a nurse working in the ICU reported, \u201cMany people die without family present. The sorrow that comes with it hurts the nurses mentally.\u201d It is encouraging to see how innovative and creative many nurses are becoming during this pandemic as they find ways to involve families. Despite being committed to the care of the ill person, they assure families that their family member is being cared for and will not die alone. They are sometimes able to connect family members to each other using new technology. Mobile phone or video conference calls made by the nurse allow family members to \u201csee\u201d the patient in the ICU or in the nursing home.Our concerns also focus on the long-term implications for patients and their families; how will they cope once the coronavirus is under control? How will they be able to resume normal life again? Individuals and families are often flexible and resilient and manyFor example, how will patients and families recover after a long period of intensive care? Research has documented that many patients experience many physical and psychological problems after such a long period of ventilation, even after discharge to their home environment . We alsoWe also hold our hearts out for families who have lost someone without being able to say goodbye and without being able to be present in the final dying phase of their loved one. How will we assist these families to cope with their loss and complex grief?Our concerns also go out to the health care professionals, especially nurses and doctors, in the aftermath of this COVID-19 crisis. They too will need support in recovering from their suffering and distress.The good news is that there is compelling evidence that our family nursing assessment and intervention skills can assist families to heal. This pandemic makes us more deeply aware of the important role of family in the lives of patients, clients, and residents. We anticipate that this increased awareness will help us advocate even more strongly for the importance of family nursing during and after this coronavirus crisis. Rightfully, a great amount of money and resources are now being spent to fight the COVID-19 virus. But lives saved must also be lives worth living afterwards.We believe that family nursing knowledge, developed over the last 40 years, unequivocally offers the necessary skills to help families recover and heal from the expected and unexpected long-term consequences of this pandemic , 2017. TFAMily health in Europe-Research in Nursing (FAME-RN) group:This Guest Editorial has been written by members of the m.l.a.luttik@pl.hanze.nl ORCID: https://orcid.org/0000-0002-7853-9773Marie Louise A. Luttik, PhD, RN, Professor in Family Nursing & Family Care, Hanze University of Applied Sciences, Research Group Nursing Diagnostics, Groningen, the Netherlands. Email: romy.mahrer@ns-c.ch ORCID: https://orcid.org/0000-0002-8587-3817Romy Mahrer-Imhof, PhD, Professor for Family-Centered Care, Nursing Science & Care Limited, Winterthur, Switzerland; Visiting Professor, Department of Clinical Research, University of Southern Denmark, Denmark. Email: cristina.garciavivar@unavarra.es ORCID: https://orcid.org/0000-0002-6022-559XCristina Garc\u00eda-Vivar, PhD, RN, Senior Associate Professor, Faculty of Health Sciences, Public University of Navarre; Researcher, IdiSNA, Navarra Institute for Health Research, Spain. Email: anne.broedsgaard.madsen@regionh.dk ORCID: https://orcid.org/0000-0002-5029-9480Anne Br\u00f8dsgaard, PhD, RN, Senior Researcher, Department of Pediatrics and Adolescent Medicine, Copenhagen University Hospital Amager Hvidovre; The Capital Region of Denmark & Section for Nursing, Department of Public Health; The Faculty of Health, Aarhus University, Denmark. Email: Karin.dieperink@rsyd.dk ORCID: https://orcid.org/0000-0003-4766-3242Karin B. Dieperink, PhD, RN, Associate Professor, Head of research, Family focused health care research Center (FaCe) and Vice Head, Department of Clinical Research, University of Southern Denmark; Department of Oncology, Odense University Hospital, Denmark. Email: lorenz.imhof@ns-c.ch ORCID: https://orcid.org/0000-0001-8441-3598Lorenz Imhof, PhD, Professor for Community-Based Care, Nursing Science & Care Limited, Winterthur, Switzerland. Email: boestergaard@health.sdu.dk ORCID: https://orcid.org/0000-0002-9094-8123Birte \u00d8stergaard, PhD, Associate Professor, Department of Clinical Research, University of Southern Denmark, Denmark. Email: eks@hi.is ORCID: https://orcid.org/0000-0003-1284-1088Erla Kolbrun Svavarsdottir, RN, PhD, FAAN, Professor, School of Health Sciences, Faculty of Nursing, University of Iceland, Iceland. Email: hanne.konradsen@regionh.dk ORCID: https://orcid.org/0000-0002-7477-125Hanne Konradsen, PhD, Professor, Herlev and Gentofte Hospital, Department of Gastroenterology, Denmark, Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark; Associate Professor, Department of Neurobiology, Care Sciences and Society, NVS, Karolinska Instituttet, Sweden. Email:"} +{"text": "Nature Communications 10.1038/ncomms15871, published online 21 June 2017.Correction to: The original version of this Article contained an error in the author affiliations.Song-Guo Zheng was incorrectly associated with the \u2018Center for Clinical Immunology, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, China.\u2019This has not been corrected in the PDF and HTML versions of the Article."} +{"text": "In \u201cRisk-Taking Behaviors and Adherence to HIV Pre-Exposure Prophylaxis in Users of Geosocial Networking Apps: Real-World, Multicenter Study\u201d :e22388) the authors noted one error.A typographical error in the originally published version of the paper rendered the second half of the article title as:Real-Word, Multicenter StudyThis has been corrected to:Real-World, Multicenter StudyThe correction will appear in the online version of the paper on the JMIR Publications website on November 9, 2020, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "HHEX variant rs1111875 was successfully demonstrated for the first time in a South Indian population.In this issue, there are eight articles: six Original Articles, one Application Note, and one article in the category of Genome Archives. The first original article, by the group of Kim et al. proposed a novel method for the analysis of single-cell RNA sequencing. Specifically, their article described a semi-automatic method that calculates a normalized score for each cell type based on a user-supplied cell-type\u2013specific marker gene list. Second, Liju et al. identified associations of genetic variants with early-onset of type 2 diabetes in a South Indian population. Although the sample size was not large, the association of the Streptomyces coelicolor A3(2), which is a Gram-positive soil bacterium known for the production of several antibiotics used in various biotechnological applications. The application of several bioinformatics tools successfully revealed the characteristics of this hypothesized protein from the genome of S. coelicolor, including its structure, function, and homologous proteins. The fourth article, by the group of Sa et al. , presented a comparative gene expression analysis regarding seed pigmentation in maize by comparing differently expressed genes from three inbred lines, including a pigment-accumulating seed type (CM22) and non-pigmented seeds (CM5 and CM19).The third article, by Ferdous et al. , provided a molecular characterization and functional annotation of a hypothesized protein of Recent active research efforts have produced many articles on the coronavirus disease 2019 (COVID-19) pandemic. In this issue, there are two articles related to COVID-19. The first one, by Sohpal presents a computational analysis of the genome of severe acute respiratory syndrome coronavirus 2 with the genome of the Middle East respiratory syndrome-related coronavirus. Using molecular evolutionary genetic analysis (MEGA) from the National Center for Biotechnology Information (NCBI) for statistical analysis, the best substitution pattern and transition/transversions (R) were compared. The final Research Article was by Apio et al. , which presents the 95% confidence intervals for the SARS-CoV-2 antibody retention rate for the Korean. The most conservative 95% confidence interval estimation showed that as of 00:00 on September 15, 2020, there were at least 32,602 undetected cases of COVID-19 in Korea.Cirrhinus reba, collected from the Khulna region of Bangladesh.The one article in the Genome Archives categories, by Islam et al. presented the sequencing and annotation of the complete mitochondrial genome of a threatened labeonine fish, Genomics & Informatics Annotation Hackathon (GIAH) event, focusing on improving earlier versions of the full-text corpus of Genomics & Informatics by semi-automatically detecting and correcting PDF-to-text conversion errors and optical character recognition errors.The Application Note, by Park and his students described the initiation of the first"} +{"text": "Royal Society Open Science manuscripts is dedicated to Emerging trends in one- and two-dimensional nanomaterials and features excellent contributions from world leaders in the field. Specifically, the issue deals with recent progress in the rapidly emerging topic of nanomaterials, focusing specifically on the synthesis, characterization and application of so-called nanocarbon materials . This timely collection of diverse research covers a wide variety of applications related to electronics, energy and quantum computing. Here, a brief overview of one- and two-dimensional nanomaterials is given, with some of the featured contributions highlighted with the aim of inspiring readers and stimulating further advances the field.This themed collection of et al. [Carbon is the fourth most abundant element in the universe and carbon compounds form the basis of all known life on Earth. In the last few decades, new carbon allotropes, beyond diamond and graphite, have emerged, where carbon occupies diverse structures and configurations. The advent of one-dimensional carbon nanotubes (CNTs) by Iijima in 1991 [et al. , and theet al. . It is wet al. [et al. [The vast majority of contributions to this issue are devoted to carbon-based nanomaterials. In this context, the energetics of perylene encapsulated within a metallic CNT and a semiconducting CNT were computationally examined by Nagasawa et al. . The autet al. has stud [et al. .Recent progress in developing copper/CNT composites towards the realization of lighter alternatives to copper is reviewed by Sekiguchi & co-workers . The autet al. [et al. [Transmission electron microscopy (TEM) imaging has been employed to monitor nanosized particles and CNT growth in a statistical manner by Xiang and Maruyama . Furtheret al. to local [et al. present et al. [60-containing C60 tetramer based on the quadruple 1,3-dipolar cycloaddition reaction, working towards the realization of a scaffold for measuring multiple qubit\u2013qubit interactions. A further paper on fullerenes by Aoyagi et al. [60 with Li+ inside the cage. The work aids our understanding of the electrostatic and thermal properties of an encapsulated lithium cation. The isolation of CF3-functionalized Gd@C74 and its structural characterization based on single-crystal X-ray diffraction is reported by Nakagawa & co-workers [Finally, on the subject of fullerenes, Macpherson et al. have achi et al. reports -workers Royal Society Open Science for their continuous support for the realization of this issue devoted to Emerging trends in one- and two-dimensional nanomaterials.The guest co-editors of this themed issue, Professor Hisanori Shinohara, at the Department of Chemistry, Nagoya University, Japan; Professor Young Hee Lee, at the Department of Energy Science & Department of Physics, Sungkyunkwan University, Korea; and Dr Nikos Tagmatarchis, at the Theoretical and Physical Chemistry Institute, National Hellenic Research Foundation, Greece, wish to thank all contributors for their enthusiasm to participate. We would also like to express our gratitude to the Royal Society of Chemistry team and in particular for the staff of"} +{"text": "In the published article, an author name was incorrect as Yasir Syed. The correct name is Yasir Ahmed Syed.In the original article, we neglected to include the funder Cambridge NIHR Brain Injury MedTech Cooperative, NIHR Clinician Scientist Award CS-2015-15-023 to Mark Kotter. The updated Funding statement appears below.We gratefully acknowledge support by the Cambridge NIHR Brain Injury MedTech Cooperative. MK was funded by a NIHR Clinician Scientist Award CS-2015-15-023.This report is independent research arising from a Clinician Scientist Award, CS-2015-15-023, supported by the National Institute for Health Research. The views expressed in this publication are those of the authors and not necessarily those of the NHS, the National Institute for Health Research or the Department of Health.The authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Parasteatonyssus nyctinomi (Chiroptera: Molossidae) captured in the Kunene region of Namibia (southern Africa). This is the first report on P. nyctinomi in the country.Sixty-four individuals of a macronyssid mite, Parasteatonyssus cornutus , Parasteatonyssus hoogstraali , Parasteatonyssus jayanti Advani, Vasirani, 1981, Parasteatonyssus lingeraji and Parasteatonyssus nyctinomi have been recorded previously in Africa.Species in the family Macronyssidae Oudemans 1936 include parasites of mammals, birds and reptiles. The more derived macronyssidae, subfamily Ornithonyssinae Lange 1958, are a relatively uniform group characterised by some specific morphological peculiarities Radovsky . Genus PP. nyctinomi from the Egyptian free-tailed bat, Tadarida aegyptiaca (Chiroptera: Molossidae), in Namibia.This article presents new findings about \u03bcm).The research was conducted on Namib Desert bats and their ectoparasites in the Kunene region of northwestern Namibia. Bats were captured from 06 December 2016 to 04 April 2017 by deploying mist nets, and captured individuals were examined intensively for ectoparasites. Bats were morphologically identified using taxonomic descriptions given by Monadjem et al. . All ectVoucher specimens mounted on slides have been deposited at the collection centre of the University of Tyumen\u2019s Museum of Zoology, Tyumen, Russia.All applicable institutional, national and international guidelines for the care and use of animals were followed. Fieldwork was conducted with in accordance with the guidelines of Colorado State University\u2019s Institutional Animal Care and Use Committee (Protocol #15-6140A) and the Ministry of Environment and Tourism, Republic of Namibia (research/collecting permits #2122/2016 and #2225/16).T. aegyptiaca, individuals in three localities: Spaarwater Pos , Sesfontein and Hoanib Skeleton Coast Camp .Sixty-four specimens of macronyssid mites were obtained from four Egyptian free-tailed bat, P. nyctinomi (Zumpt, Patterson, T. aegyptiaca. The Egyptian free-tailed bat is widely distributed throughout Africa, except parts of the northwest and east through Arabia and the Middle East to southern Asia as far east as Bangladesh and south to Sri Lanka (Skinner, Chimimba, P. nyctinomi is a specific ectoparasite of T. aegyptiaca is true, we can expect new records of the mite in the Palearctic part of Africa, Arabia and India where T. aegyptiaca is endemic. The lack of records is most likely because of collection difficulties. Because of a lack of knowledge about the life cycle of P. nyctinomi, more studies are necessary to determine the principal host and geographic distribution of this parasite species.This is the first report from Namibia on"} +{"text": "Scientific Reports 10.1038/s41598-019-42782-x, published online 23 April 2019Correction to: The original version of this Article contained errors in the spelling of the authors Fiona C. Brown, Stephen M. Jane and David J. Curtis which were incorrectly given as Fiona Brown, Stephen Jane and David Curtis respectively.These errors have now been corrected in the PDF and HTML versions of the Article, and in the accompanying Supplementary Information file."} +{"text": "Background: The population is aging much faster in China than other low- and middle-income countries. With the accelerated aging of the population, incidence and disease burden of age-related diseases have also continued to increase. Exploring the burden of age-related diseases is crucial for early disease prevention, assessing the extent of population aging, and achieving the goal of healthy aging.Methods: We used the dataset from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD), and selected data on incidence, prevalence, and disease burden in China, in 1997, 2007, and 2017. We classified age-related diseases, which were defined as diseases in which the incidence rate increased quadratically with age in the adult population. Additionally, we described the changes in age-related diseases during the study period by different GBD categories. It also measured changes in the age-related disease burden in our study period, including disability-adjusted life years (DALY), years of life lost (YLL), and years lived with disability (YLD). Finally, we compared the differences in the age-related disease burdens for men and women.Results: Among the 293 diseases listed in the GBD study, 69 in 2017, 78 in 1997 and 72 in 2007 were identified as age-related diseases. More than half of the age-related diseases belonged to non-communicable diseases (NCDs) in our study period. The rate of age-standardized age-related disease burden decreased between 1997 and 2017. DALYs decreased by 24.89% for non-age-related diseases and by 50.15% in age-related diseases from 1997 to 2017. The age-related disease burden of men was higher than that of women; we found a decreasing trend, with \u221246.23% in men and \u221254.90% in women.Conclusions: Comparing characteristics of the aging population in China and the world, we found that China does not have the typical disease characteristics of aging society. Currently, China faces the dual threat of NCDs and communicable diseases, and NCDs account for the vast majority of the age-related disease burden. Our health systems should focus on disease prevention and early detection among the entire population, instead of treatment. Further studies should focus on reducing the duration and severity of morbidity in later life. According to the report of World Health Organization (WHO), the global elderly population is expected to increase from 12 to 22% by 2050 . It is hWHO has developed a new concept of healthy aging as being much more than the absence of disease . Based oHealth problems in older age go beyond what can be caused by disease alone, such as impairments in cognition, mood, and physical performance . Thus, mIn our study, we attempted to screen out age-related diseases in the Chinese population by using GBD data, and compared the changes in the types of age-related diseases in 1997, 2007, and 2017 and analyzed the burden of age-related and non-age-related diseases during those 3 years. In this paper, to improve current understanding of the health problems faced by older adults, we analyzed the characteristics of age-related diseases in China. Furthermore, we analyzed the types and changing trends of age-related diseases to provide a basis for targeted interventions, such as early disease prevention and health management among older populations. Our findings could help clarify the sequencing and effectiveness of interventions, as well as develop design policies to support healthy aging in different contexts.http://vizhub.healthdata.org/gbd-compare/). To compare the trends over two decades, we analyzed the related data in 1997, 2007, and 2017. The GBD study design and methods have been described in previous literature and their components, years of life lost (YLLs), and years lived with disability (YLDs) for 293 diseases found in adults in China , non-communicable diseases (NCDs), and injuries. Since many previous studies , 24 haveIn our study, we used the same approach as the previous study performed by Chang et al., to screen for age-related diseases and their burden in the Chinese population; meanwhile, we defined age-related diseases as diseases with incidence rates that increased quadratically with age among adult population , 25\u201327. Age-related disease burden was considered the sum of all DALYs for age-related diseases in adults. We analyzed the age-related disease burden, which is the proportion of age-related disease burden out of the total health burden. For cross-period comparisons, we also analyzed the age-standardized rates per 1,000 adults, which were adjusted for population size and age structure.DALYs are a measure of overall disease burden, expressed as the cumulative number of years lost due to ill health, disability, or early death. One DALY can be thought of as one lost year of healthy life. The sum of these DALYs across the population, or the burden of disease, can be thought of as a measurement of the gap between current health status and an optimal health, in which the entire population lives to an advanced age, free of disease or disability.DALYs for a disease or health condition are calculated by adding the YLLs due to premature mortality in the population and the YLDs for people living with the health condition or its consequences. YLLs are calculated by multiplying the number of deaths associated with the condition by the remaining life expectancy. YLDs are calculated by multiplying the number of cases with a certain health outcome by the weight of the specified disability.All analyses were conducted by Stata version 12.0. To understand the relationship between age and disease prevalence at the population level, disease-specific data for both genders were summarized. Unless otherwise stated, all rates were expressed as age-standardized for the GBD reference population. This study followed the Guidelines for Accurate and Transparent Health Estimates Reporting (GATHER) 20 recommendations.We used the same techniques found elsewhere in GBD research design to propagate uncertainty. The final estimate was calculated using an average estimate of 1,000 samples, while 95% of the uncertainty interval (UI) was determined based on the 25th and 975th bit-sorted values of all 1,000 samples.To verify the stability of the results, we excluded incidence data for ages 80 and above for sensitivity analysis. The analysis showed that age-related disease outcomes between 25 and 80 years were consistent with those identified in the adults ages 25 and above.In 2017, 69 (23.5%) age-related diseases were identified among the 293 GBD causes; 78 (26.6%) and 72 (24.6%) were identified in 1997 and 2007, respectively. In 2017, among the 69 conditions, 14 (20.29%) were CMNNs, 7 (10.14 %) were injuries, and 48 (69.57%) were NCDs. No significant change or injury was observed in the age-related CMNNs during the study period, but the number of age-related NCDs decreased. Additionally, we also found that several emerging age-related diseases, such as other pneumoconiosis, acute glomerulonephritis, age-related macular degeneration, and near vision loss increased in 2017 .A total of 248.34 DALYs were due to the age-standardized, age-related disease burden; however, 400.41 DALYs were caused by non-age-related disease. In terms of proportion, age-related disease burden accounted for 38.28% (248.34/648.75) of the total disease burden. Causes of non-age-related disease burden included neoplasms (19.91%), injuries (13.12%), neurological disorders (11.91%), CMNNs (11.59%), and musculoskeletal disorders (10.39%). For age-related diseases, the burden was primarily due to cardiovascular disease (44.16%), other NCDs (12.71%), chronic respiratory disease (12.01%), injuries (7.44%), and neoplasms (6.72%). The burden of age-related disease was greater in the cardiovascular, chronic respiratory, and other chronic disease groups than in the non-age-related group .In 2017, the burden of disease in China was 648.81 DALYs, of which 59.40% were caused by death and 40.60% by disability . YLLs, YThe data of different years were segregated separately for gender. The disease burden of men was higher than women. From 1997 to 2017, the proportion of age-related disease burden in all populations showed decreasing trend (\u221246.23%), which was more apparent among women (\u221254.90%) .Our study showed that the number of age-related diseases decreased from 1997 to 2017, with more than half classified as NCDs, such as cardiovascular disease, chronic respiratory disease, and other NCDs. Over the past two decades, the disease burden has declined in China, especially for age-related diseases (50.15%). The burden of age-related NCDs was higher than CMNNs and injuries in 2017. Additionally, men had a higher disease burden than women.The number of age-related diseases was less in China (69 diseases) than globally based on the results of a study performed by Angela Chang and colleagues (92 diseases) in 2017,Our main finding was that age-standardized age-related disease burden decreased from 2007 to 2017. On one hand, it may be related to the decrease of age-related diseases during the study period. On the other hand, with the development of economy and society, people's health improved, the decrease of overall disease burden is not surprising. Whereas, one of the serious problems associated with population aging is the increasing burden of NCDs, and the prevalence of NCDs will increase along with the increasing disease burden. According to current projections, China's rapidly aging population is expected to increase the burden of NCDs by at least 40% by 2030 . Thus, mThe burden of age-related disease was greater in the cardiovascular, chronic respiratory, and other chronic disease groups; those diseases need better predictors of their occurrence. It is crucial to study an intervention on how to block the causal mechanisms at an early stage. Published studies have shown that only about 25% of our lifespan is determined by our genes; the other 75% is determined by our lifestyles and the choices we make every day , 36. We The disease burden and proportion of age-related burden among all burdens was lower for women than for men, and the disease burden decreased more for women than men during our study period. The contribution to the disease burden between men and women was unequal, due to the higher prevalence of chronic disease risk factors in men . In addiIn the absence of population level aging metrics that inform longevity, health status and disease severity in China, based on the GBD database, this is the first study to explore the age-related disease burden among Chinese adults. Furthermore, we have found changes in the overall burden of age-related disease over 20 years in China. Our study has several limitations. First, while the GBD list of diseases is comprehensive, we may have missed including some diseases that are important to China's aging population. Second, we used the same methods as Chang and colleagues to identIdentifying age-related diseases is a prerequisite for providing promising therapeutic strategies to promote healthy aging in the future. Comparing the characteristics of the aging population in China and the world, we found that China does not have the typical disease characteristics of an aging society. Currently, China faces the dual threat of NCDS and communicable diseases, and NCDs account for the vast majority of the age-related disease burden. Our health systems should focus on disease prevention and early detection among the entire population, instead of treatment. Further studies should focus on reducing the duration and severity of morbidity in later life.http://ghdx.healthdata.org/gbd-results-tool.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: DH and JC contributed to the conception and design of the study. WY, JZ, and DH conducted the data reduction and analyses. YZ helped validate the disease screening. DH wrote the manuscript, guided the whole process, and reviewed the manuscript. All authors read and approved the manuscript before submission.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The World Health Organization\u2019s End TB Strategy 2016\u20132035) sets the goal of ending the tuberculosis (TB) epidemic, in line with the United Nations Sustainable Development Goals [6\u20132035 seChina has the third-highest burden of TB and second-highest burden of multidrug-resistant TB (MDR-TB) globally. Yet between 1990 and 2010, the prevalence rate of smear-positive pulmonary TB in China declined significantly . This acThe National Health Commission (NHC) of China, formerly the Ministry of Health, oversees the national program for TB and MDR-TB prevention and control in China. With support from the Bill & Melinda Gates Foundation, the NHC has, since 2009, developed an innovative program to strengthen TB control, and the China CDC led the implementation. This program, the \u201cChina National Health Commission and Gates Foundation TB Prevention and Control Project\u201d (China-Gates TB project), was carried out in three phases, from 2009 to 2019 diagnosis, treatment, and patient management. The tools included a variety of new diagnostics that were more sensitive, and prompt for diagnosis, made standardization of MDR-TB diagnostic and treatment packages more feasible, and opened the doors for use of electronic medication monitors (EMMs) to improve treatment adherence of TB patients.In February 2013, the NHC issued Decree No. 92, which describes the process of integrating TB clinical services and patient management into China\u2019s tiered health service delivery system. At the same time, basic medical insurance schemes were expected to be a primary source to pay for TB care in the hospitals. With the transition to a more integrated and collaborative TB control model in China, Phase II of the project (2012\u20132015), which was implemented in three prefectures in eastern, central, and western provinces of China, demonstrated how to ensure that innovative diagnostic, treatment, and financing approaches for all TB patients could be effectively adapted to the new model. Key findings from Phases I and II were published in several peer-reviewed international journals \u20137.Based on the achievements of Phases I and II, Phase III was kicked off in 2016 in the provinces of Zhejiang, Ningxia, and Jilin. This phase aimed to expand the comprehensive model of TB control at the provincial level and introduce new policies, technologies, and approaches to cope with the challenges identified in the previous phases and thus improve the performance of TB control in China. Phase III centered on three domains. The first domain focused on strengthening the TB care delivery system, including (a) planning, policy development, and advocacy to support scale-up of the comprehensive TB control model through local government commitment; (b) capacity-building of TB care professionals through innovative training approaches; and (c) developing a new TB surveillance and information system. The second domain focused on improving the quality of TB care, including the (a) upgrade of laboratory capacity for TB and MDR-TB diagnostics; (b) standardization of diagnosis, treatment, and patient management of TB and MDR-TB; (c) scale-up of using EMMs to improve community-based case management; and (d) introduction of new anti-TB drugs. The third domain focused on improving the mechanism for financing TB care and included (a) introducing multisource financing for TB clinical care, (b) exploring the reform of payment methods for TB clinical care, and (c) simplifying the process of medical assistance for patients who are eligible to benefit.Duke Global Health Institute and Duke Kunshan University (a Chinese-American partnership of Wuhan University and Duke University), as a third party, led the monitoring, learning, and evaluation of Phase III of the China-Gates project. The evaluation team consisted of collaborators from Peking University, Fudan University, Zhejiang University, and China CDC. The evaluation team undertook performance-based monitoring, using routine data from all counties of the three project provinces and an in-depth site evaluation in the selected prefectures in each province. For each province, two prefectures were selected, representing a variety of socioeconomic levels. Within each prefecture, two counties were selected using the same criteria; in total, six prefectures and twelve counties were involved in the site evaluation. The team applied both quantitative and qualitative methods for the baseline and final evaluation. Quantitative data collection included a patient survey with TB and MDR-TB patients, a survey with TB professionals, an institution-based survey for TB-designated hospitals and local CDCs, as well as routine data from the health information system in the TB-designated hospitals, medical records of TB and MDR-TB patients, and surveillance data on TB and MDR-TB diagnosis, treatment, and case management. Qualitative data included data extracted from local policy documents; qualitative interviews with decision-makers and key informants from the local health authority, health insurance agency, CDC, TB-designated hospitals, and focus group discussions with TB care providers and TB and MDR-TB patients; site observations; and field notes.The quality of diagnosis and treatment of TB and MDR-TB significantly improved in the project provinces, which is, to a great extent, attributable to the availability and accessibility of new diagnostic technologies and standardization of treatment procedures. There was a notable increase of bacteriologically confirmed pulmonary TB cases and a rise in the proportion of TB and DR-TB patients receiving the recommended diagnostic services and clinical treatment at the time of project final evaluation in 2019 compared to the baseline study in 2016. However, irrational use of second-line anti-TB drugs for TB treatment remained, and varied in magnitude, across the study sites. The qualitative results reveal that the achievements were mainly attributed to successful scale-up of the new rapid diagnostic technologies as well as training and monitoring of TB clinical services. Meanwhile, the qualitative findings raise concerns on the sustainability and affordability of applying new diagnostic technologies. In terms of appropriate application of new anti-TB drugs, the safety profile of bedaquiline-containing treatment for patients with MDR-TB and extensively drug-resistant TB (XDR-TB) was examined . The resA mechanism of multisource financing for TB and MDR-TB treatment has been introduced in the project provinces. However, the introduction was initially delayed, and thus many households with a DR-TB patient suffered from a catastrophic health expenditure at the time of project final evaluation. Strengthening cooperation among multiple sectors and improving the accountability of different government agencies will be the key to ensure significant progress toward alleviating the financial burden of TB and MDR-TB patients . In addiCapacity-building of TB professionals through e-learning and reform of the TB information system are two approaches to strengthening the TB care delivery system. The scale-up of e-learning for continuing TB medical education shows that TB clinical doctors and primary health care providers benefited from e-learning and applied what they learned in routine practice, while, the training for public health physicians needed improvement. Challenges in promoting of e-learning in continuing medical education included unmet learning needs, weak leadership, unfriendly environment, and lack of incentives to TB professionals participating in the e-learning . In addiWith the support of local government in the project provinces, the scale-up of new diagnostics and training and monitoring of TB clinical services contributed to improvement in the quality of diagnosis and treatment of TB and MDR-TB. Yet coordination of multisource financing for TB care is the most challenging component due to the various interests of stakeholders, weak leadership, and poor engagement of multiple sectors. Knowledge gained from introduction of e-learning for TB training and a pilot of reform of the TB information system both indicated the need for investment in infrastructure and human resources.Furthermore, the COVID-19 pandemic has had a devastating effect on TB responses globally. Health system overload from treating COVID-19 cases and preventing transmission has, to some extent, resulted in a decrease of, or limit on, anti-TB services delivery. Similarly, the economic recession caused by the pandemic has had a negative impact on the generation of health resources. Given the potential impact of COVID-19 on TB control in the near future, improving performance of TB control should be in line with government efforts toward the development of effective universal health coverage in China.The authors hope that the experience and findings from the China-Gates TB project can contribute to successful achievement of innovative approaches for TB control in other low- and middle-income countries with a high TB burden."} +{"text": "Nature Communications 10.1038/s41467-019-12438-5, published online 27 May 2020.Correction to: The original version of this Article omitted from the Genome Aggregation Database consortium the member Marquis P. Vawter, from the Department of Psychiatry & Human Behavior, University of California Irvine, Irvine, CA, USA. Additionally, the following was added to the Author Contributions: \u2018All authors listed under The Genome Aggregation Database Consortium contributed to the generation of the primary data incorporated into the gnomAD resource\u2019.This has been corrected in both the PDF and HTML versions of the Article."} +{"text": "The COVID-19 pandemic has disproportionately infected older adults who are also at higher risks of developing severe health complications and having case fatality if infected. In particular, older adults in residential and community-based facilities are at greater risks of contracting COVID-19, in part due to close proximities and frequent human interactions in these facilities. In Singapore, the Ministry of Health and its implementation agency, the Agency for Integrated Care (AIC), together with long-term care service providers, have jointly developed a number of precautionary measures against COVID-19. This paper defines and describes Singapore\u2019s response in terms of the \u2018ABC\u2019 of COVID-19 safety measures for facilities, namely, safe Access, Behaviors, Compounds . These included infection prevention and control measures, access to PPE, distancing and zoning measures, suspension of visitors, alternative accommodation for long-term care workers, and testing to monitor people with long-term care needs and care workers. Incident response teams to support facilities providers in responding to COVID-19 infections were also swiftly set up. The outreach arm of AIC, the Silver Generation Office, further provided information and services support to older people during the pandemic. We share these measures as a set of practices and learning points for other countries undergoing the current pandemic and for future references."} +{"text": "In the original version of the article, the co-author would like to add to the acknowledgements section to highlight their funding stream (EPSRC). The revised acknowledgements is given below.Orcid number of last author Emma M. Clark inadvertently associated with the first author Amir Jamaludin in the original publication. This has been corrected in this erratum.https://www.bristol.ac.uk/alspac/external/documents/grant-acknowledgements.pdf. This research was specifically funded by the British Scoliosis Research Foundation. This publication is the work of the authors, and EC will serve as guarantor for the contents of this paper, which do not reflect the views of the ALSPAC executive.We are extremely grateful to all the families who took part in this study, the midwives for their help in recruiting them, and the whole ALSPAC team, which includes interviewers, computer and laboratory technicians, clerical workers, research scientists, volunteers, managers, receptionists, and nurses. The UK Medical Research Council and Wellcome Trust (Grant Ref: 102215/2/13/2) and the University of Bristol provide core support for ALSPAC. A comprehensive list of grants funding is available on the ALSPAC website Funding for authors based at the University of Oxford was via an EPSRC Programme Grant Seebibyte (EP/M013774/1)."} +{"text": "The brain is a complex, multiscale dynamical system composed of many interacting regions. Knowledge of the spatiotemporal organization of these interactions is critical for establishing a solid understanding of the brain\u2019s functional architecture and the relationship between neural dynamics and cognition in health and disease. The possibility of studying these dynamics through careful analysis of neuroimaging data has catalyzed substantial interest in methods that estimate time-resolved fluctuations in functional connectivity . At the same time, debates have emerged regarding the application of TVFC analyses to resting fMRI data, and about the statistical validity, physiological origins, and cognitive and behavioral relevance of resting TVFC. These and other unresolved issues complicate interpretation of resting TVFC findings and limit the insights that can be gained from this promising new research area. This article brings together scientists with a variety of perspectives on resting TVFC to review the current literature in light of these issues. We introduce core concepts, define key terms, summarize controversies and open questions, and present a forward-looking perspective on how resting TVFC analyses can be rigorously and productively applied to investigate a wide range of questions in cognitive and systems neuroscience. Even when sitting quietly in a dark room, the brain is active, yielding a constant stream of thoughts and ideas, along with changes in awareness, Functional connectivity (FC) analyses of resting fMRI (rfMRI) data allow researchers to noninvasively estimate patterns of interregional neural interactions. An integral component of modern neuroimaging research, FC is traditionally calculated over an entire scan or experimental condition , but recent years have seen rapidly growing interest in studying time-resolved fluctuations in FC a small number of replicable patterns of connectivity that recur across or within individuals , it is critically important to ensure that one distinguishes between the method and the target theoretical properties we wish to infer . Failure to do so commits the logical fallacy of confusing the map for the territory . These interactions have many different properties we may be interested in, such as their direction , directness , and timing . While most FC methods applied to BOLD fMRI data are limited in the extent to which they can provide information about the exact structure of the underlying causal graph, they nonetheless constrain the space of possible network configurations Are rfMRI time series statistically consistent with functional connectivity that truly varies in time? (2) What is the biological basis of BOLD TVFC ? (3) What (if any) is the cognitive and behavioral relevance of resting BOLD TVFC? We begin with a survey of the current landscape of analytic and modeling approaches for studying BOLD TVFC, and then proceed to address each of the three questions outlined above. First, we review methodological considerations and statistical challenges for studying TVFC in fMRI. Second, we review the literature on the physiological basis of BOLD TVFC. Third, we provide an in-depth discussion of the cognitive and behavioral relevance of BOLD TVFC, including evidence both for and against this proposition. Subsequent sections highlight experimental approaches that may help adjudicate questions about the cognitive relevance of TVFC, and briefly review strategies for cleaning rfMRI data to mitigate the impact of potential confounds on TVFC analyses. We conclude by suggesting ways that the TVFC research community can continue to advance this exciting field and help facilitate consensus on controversial issues.between states.Approaches to studying functional connectivity in fMRI data can be considered along a spectrum of temporal resolution. On one end, some methods assume that the dependence structure between regions is constant over an arbitrarily long time window ; on the other end are methods that can estimate time-resolved FC at each individual time point . In between are methods that aim to discover discrete, temporally contiguous functional connectivity states characterized by their interregional dependence structure . In these state-based models, the dependence structure changes only when moving Another important property of methods used to study TVFC is the extent to which they consider the temporal ordering of the observed data points. Some approaches directly leverage the information in this ordering directly from the observed BOLD data perspectives on the data, and a full understanding of the factors giving rise to TVFC and their relationship to cognition and behavior will likely necessitate integrating knowledge gained through the application of a wide variety of methods (see Box 3). Some approaches make minimal (or no) explicit assumptions about the underlying biology, while others seek to model the biophysical parameters directly. Improved biological specificity is often accompanied by greater model complexity and more extensive explicit model assumptions. That said, methods that directly model the observed BOLD signal can also be highly statistically articulated and come with their own assumptions that are often just as strong as assumptions made by biophysical models.Highly articulated \u201cdata-driven\u201d models be drawn from each method. Below, we briefly outline how four broad classes of TVFC methods can be used to expand our understanding of brain function.Time-resolved estimates of functional connectivity: Empirical estimates of time-resolved functional connectivity allow scientists to explore how the strength of interregional coupling varies over time. These estimates form the basis of empirical studies of TVFC. In their most basic form , they can provide insight into the trajectories by which static (\u201ctime-averaged\u201d) FC is realized. Time-resolved estimates also allow for fine-grained evaluations of the relationship between FC and ongoing cognition, as well as how summary measures may be related to phenotypic traits in health and disease.Models of states and transitions: Many empirical studies of TVFC also seek to estimate transient \u201cbrain states\u201d and their transitions. In this paradigm, each state describes a different pattern of whole-brain activity or functional connectivity. Different models impose varying constraints on the estimated states, such as whether they manifest in isolation (one state per time point) or in combination (a mix of states at each time point). The dynamics of these states can provide a detailed portrait of how functional relationships reorganize through time. Formal model selection and comparison allows for the evaluation of which models best describe the observed data, and thus permit adjudication of competing hypotheses about data-generating processes.Comparison to surrogate (null) data: Insight into the dynamical properties of a system can also be achieved by comparing observed data to surrogate data that lack a particular statistical feature of interest. For example, one can generate surrogate \u201cnull\u201d time series that have the same low-order features as empirical data but lack a higher order feature proposed to exist in the real data . The strength of this approach is that it draws from a rich existing literature on time series analysis and enables testing of specific hypotheses about the dynamical properties of an observed time series. Care must be taken to ensure that the tests undertaken are sufficiently narrow and are interpreted as such. For example, claims should be made about the presence or absence of a particular statistical feature rather than \u201cdynamic\u201d FC in general, as \u201cdynamic\u201d phenomena can exist under a wide range of conditions.Modeling of nonlinear brain dynamics:Unlike the three approaches above that begin with empirically measured BOLD data, it is also possible to instead begin the study of TVFC by constructing a detailed biophysical model of the underlying processes thought to give rise to TVFC. With appropriate model fitting and tuning, it is possible to invert the observed data into a generative model, and then study the complex dynamical properties of that model that would normally be obscured by the measurement process. Having established a model of the dynamical processes underlying the observed data, researchers can undertake detailed mechanistic investigations of complex neural dynamics and their relationship to BOLD TVFC.Before diving into questions about the biological basis and cognitive relevance of resting BOLD TVFC, we must first ask whether there is statistical evidence for this phenomenon: Does functional connectivity estimated from resting BOLD fMRI actually vary over time? In this section, we discuss the importance of testing TVFC estimates against null models, review the role of Any method designed to estimate TVFC will inevitably return time-resolved estimates of functional connectivity that vary to some degree with time estimate a series of correlations, it can be useful to think of these values as \u201crepeated samples\u201d of correlations across time. From this perspective, the key question being asked when evaluating TVFC estimates is whether each sample was drawn from the same distribution (static FC) or from distinct distributions (TVFC). If we choose a small window size, the correlation coefficient will be based on few data points; this gives rise to larger sampling variability. Thus, short window lengths may give rise to signals that show compellingly \u201cdynamic\u201d changes in correlation across time, even if the FC is actually static , evaluations of sensitivity typically make use of simulated data containing a known TVFC signal of interest . A variety of simulation tools are available to help researchers evaluate how TVFC methods perform under a range of different data-generating conditions . However, the BOLD signal is a noisy, indirect measure of underlying neural activity, and the sluggish hemodynamic response places a fundamental limit on the temporal resolution of TVFC estimated from fMRI data. It is well established that the shape of the hemodynamic response function varies across brain areas also contribute substantially to measures of functional connectivity and TVFC that multiple, potentially dissociable neurophysiological processes simultaneously contribute to time-varying BOLD activity and functional connectivity, and there is good reason to believe that the heterogeneity of electrophysiological frequency bands reported as being associated with BOLD static FC and TVFC is not merely artifactual or due to experimental variability. Different bands of electrophysiological activity likely reflect distinct neurophysiological processes often relate to cognition in nontrivial ways . In addition, task-related changes in estimates of FC can be driven by relatively trivial changes in coordinated activity, such as those resulting from stimulus-induced coactivation . A growing number of studies have observed synchronized fluctuations in TVFC as participants watch a movie or listen to a story prior to analysis of fMRI data, and this is particularly true when working with data acquired during rest. While most standard rfMRI preprocessing steps . This can be done by directly comparing potential confounding factors between groups, or by testing for a correlation between potential confounds and TVFC. It is also possible to test for residual confounding after preprocessing and compare these residual estimates between groups independent of arousal must therefore measure and account for the presence and influence of these fluctuations. Although it can be difficult to track subtle fluctuations in arousal, measurements of pupil diameter of TVFC during rest : Given that patterns of static FC are so similar across a wide range of behavioral states, why should we expect to see fluctuations in FC during periods of rest, when overt behavior remains unchanged? The key to resolving this paradox may involve recognizing that a given value of a summary measure can be realized by multiple different arrangements of the underlying data. In the context of FC, this means that the same pattern of static FC may result from different spatiotemporal patterns of underlying TVFC. Further, because TVFC fluctuations unfold over time in a particular order, the same distribution of TVFC patterns can have very different temporal profiles . Brain dynamics unfold over time in a particular sequence, and it is therefore important to go beyond simply identifying FC patterns at high temporal resolution: To further our understanding of brain dynamics, cognition, and behavior, we must also consider the temporal aspects of TVFC fluctuations . It is not enough to know It is our hope that this paper can serve as not only a review of the current state of the field, but also a blueprint for future work. TVFC analyses of BOLD fMRI and other types of neuroimaging data have the potential to help answer some of the most compelling open questions in cognitive and systems neuroscience. TVFC analyses of intrinsic brain activity recorded at rest are fast becoming a key tool for researchers seeking to identify fundamental principles of macroscale brain dynamics, their spatial and temporal organization, and their relationship to underlying anatomy. Studies of resting TVFC have also begun to shed light on disordered intrinsic brain dynamics in individuals with psychiatric and neurological illness, and careful experiments using online measures and naturalistic paradigms promise to reveal fine-grained relationships between patterns of functional connectivity and cognitive, behavioral, and physiological states. At the same time, important questions remain unresolved. How much variance in resting TVFC is explainable by various contributing factors ? Precisely how sensitive is BOLD TVFC to shifts in cognition? Can we resolve \u201cspontaneous\u201d changes in mental content , or are we limited to studying more general changes in cognitive state ? Success in answering these questions will require contributions from and collaboration between researchers with a wide range of backgrounds and perspectives. With this in mind, we offer the following concrete recommendations aimed at facilitating a consensus approach for research into time-varying functional connectivity.First, we urge researchers undertaking TVFC analyses to carefully consider their choice of terminology when describing their methods and framing their results. Inconsistencies in definitions between researchers have the potential to needlessly muddy an already complicated scientific landscape. While we have proposed the term \u201ctime-varying functional connectivity\u201d as an appropriately broad label, we recognize that debates about the application of this and other terms are likely to continue. Beyond the specific case of TVFC, there is also ongoing debate about the use of \u201cfunctional connectivity\u201d to refer to methods that attempt to infer neural interactions from time series data , we recognize that science rarely proceeds in an orderly fashion. As such, it is critical that studies exploring the \u201clatter\u201d questions make clear on which untested or controversial assumptions they rest .As the field continues to move forward, the study of resting brain dynamics will benefit from both the refinement of existing TVFC methodologies as well as the use and development of complementary techniques. For example, methods capable of recovering the hemodynamic response function from rfMRI data promise to further elucidate the relationship between neural activity at rest and the observed BOLD signal. This knowledge can in turn help inform the development of models that facilitate the estimation of time-varying directed or \u201ceffective\u201d functional connectivity. As our tools and analyses continue to develop, it will also be critical to assess the impact of data quality and quantity on individual and group-average TVFC estimates.Overall, we believe that statistically rigorous, well-validated studies of resting BOLD TVFC have the potential to greatly expand our understanding of brain dynamics and their relationship to cognition in health and disease, and that collaborative, open work towards resolving outstanding controversies is the most effective and productive path forward for our field.We would like to thank Cesar Caballero-Gaudes, Peter Fransson, Lucina Uddin, Sepideh Sadaghiani, Daniele Marinazzo, and Ioannis Pappas for their comments and suggestions on earlier versions of this manuscript. We also thank Abraham Snyder, Jean-Baptiste Poline, Ted Satterthwaite, Caterina Gratton, and Nico Dosenbach for their participation in the discussion that inspired us to write this review.https://doi.org/10.1162/netn_a_00116. Python code for creating Supporting information for this article is available at Daniel J. Lurie: Conceptualization; Project administration; Supervision; Visualization; Writing - Original Draft; Writing - Review & Editing. Daniel Kessler: Conceptualization; Project administration; Supervision; Writing - Original Draft; Writing - Review & Editing. Danielle S. Bassett: Writing - Original Draft; Writing - Review & Editing. Richard F. Betzel: Writing - Original Draft; Writing - Review & Editing. Michael Breakspear: Writing - Original Draft; Writing - Review & Editing. Shella Kielholz: Writing - Original Draft; Writing - Review & Editing. Aaron Kucyi: Writing - Original Draft; Writing - Review & Editing. Rapha\u00ebl Li\u00e9geois: Writing - Original Draft; Writing - Review & Editing. Martin A. Lindquist: Writing - Original Draft; Writing - Review & Editing. Anthony Randal McIntosh: Writing - Original Draft; Writing - Review & Editing. Russel A. Poldrack: Writing - Original Draft; Writing - Review & Editing. James M. Shine: Writing - Original Draft; Writing - Review & Editing. William Hedley Thompson: Writing - Original Draft; Writing - Review & Editing. Natalia Z. Bielczyk: Visualization; Writing - Review & Editing. Linda Douw: Writing - Review & Editing. Dominik Kraft: Writing - Review & Editing. Robyn L. Miller: Writing - Review & Editing. Muthuraman Muthuraman: Writing - Review & Editing. Lorenzo Pasquini: Writing - Review & Editing. Adeel Razi: Writing - Review & Editing. Diego Vidaurre: Writing - Review & Editing. Hua Xie: Writing - Review & Editing. Vince D. Calhoun: Conceptualization; Project administration; Supervision; Writing - Original Draft; Writing - Review & Editing.http://dx.doi.org/10.13039/501100001942), Award ID: IVAN 20CH21 174081. Martin A. Lindquist, National Institute of Biomedical Imaging and Bioengineering, Award ID: R01 EB016061. Martin A. Lindquist, National Institute of Biomedical Imaging and Bioengineering, Award ID: R01 EB026549. James M. Shine, National Health and Medical Research Council, Award ID: GNT1072403. James M. Shine, National Health and Medical Research Council, Award ID: GNT1156536. William Hedley Thompson, Knut and Alice Wallenberg Foundation, Award ID: 2016.0473. Linda Douw, Society in Science, Award ID: Branco Weiss Fellowship. Linda Douw, Dutch Organization for Scientific Research, Award ID: 016.146.086. Adeel Razi, Australian Research Council, Award ID: DECRA, DE170100128. Dominik Kraft, German Research Foundation, Award ID: CRC 1193. Dominik Kraft, German Research Foundation, Award ID: INST 247/859-1. Muthuraman Muthuraman, German Research Foundation, Award ID: SFB CRC-1193. Muthuraman Muthuraman, German Research Foundation, Award ID: SFB CRC-TR-128. Vince D. Calhoun, National Institutes of Health, Award ID: R01EB020407. Vince D. Calhoun, National Institutes of Health, Award ID: P20GM103472. Vince D. Calhoun, National Institutes of Health, Award ID: P30GM122734. Vince D. Calhoun, National Science Foundation, Award ID: 1539067.Daniel J. Lurie, National Science Foundation, Award ID: DGE 1106400. Daniel J. Lurie, National Institute of Neurological Disorders and Stroke, Award ID: 1F31NS108665-01A1. Daniel Kessler, National Science Foundation, Award ID: DMS-1646108. Danielle S. Bassett, John D. and Catherine T. MacArthur Foundation. Danielle S. Bassett, Alfred P. Sloan Foundation. Danielle S. Bassett, ISI Foundation. Danielle S. Bassett, Paul Allen Foundation. Danielle S. Bassett, Army Research Laboratory, Award ID: W911NF-10-2-0022. Danielle S. Bassett, Army Research Office, Award ID: Bassett-W911NF-14-1-0679. Danielle S. Bassett, Army Research Office, Award ID: Grafton-W911NF-16-1-0474. Danielle S. Bassett, Army Research Office, Award ID: DCIST-W911NF-17-2-0181. Danielle S. Bassett, Office of Naval Research. Danielle S. Bassett, National Institute of Mental Health, Award ID: 2-R01-DC-009209-11. Danielle S. Bassett, National Institute of Mental Health, Award ID: R01-MH112847. Danielle S. Bassett, National Institute of Mental Health, Award ID: R01-MH107235. Danielle S. Bassett, National Institute of Mental Health, Award ID: R21-M MH-106799. Danielle S. Bassett, National Institute of Child Health and Human Development, Award ID: 1R01HD086888-01. Danielle S. Bassett, National Institute of Neurological Disorders and Stroke, Award ID: R01 NS099348. Danielle S. Bassett, National Science Foundation, Award ID: BCS-1441502. Danielle S. Bassett, National Science Foundation, Award ID: BCS-1430087. Danielle S. Bassett, National Science Foundation, Award ID: NSF PHY-1554488. Danielle S. Bassett, National Science Foundation, Award ID: BCS-1631550. Richard F. Betzel, Indiana University Office of the Vice President for Research, Award ID: Emerging Area of Research Initiative, Learning: Brains, Machines, and Children. Michael Breakspear, National Health and Medical Research Council, Award ID: 118153. Michael Breakspear, National Health and Medical Research Council, Award ID: 10371296. Michael Breakspear, National Health and Medical Research Council, Award ID: 1095227. Michael Breakspear, Australian Research Council, Award ID: CE140100007. Shell Kielholz, National Institutes of Health, Award ID: R01MH111416. Shell Kielholz, National Institutes of Health, Award ID: R01NS078095. Shell Kielholz, National Science Foundation, Award ID: BCS INSPIRE 1533260. Aaron Kucyi, Canadian Institutes of Health Research, Award ID: Banting Fellowship. Rapha\u00ebl Li\u00e9geois, CHIST-ERA (Click here for additional data file."} +{"text": "Nature Communications 10.1038/s41467-021-21240-1, published online 09 February 2021.Correction to: The original version of this Article contained an error in the author affiliation.Lin-Fa Wang was incorrectly associated with Thai Red Cross Emerging Infectious Diseases Health Science Centre, WHO Collaborating Centre for Research and Training on Viral Zoonoses, King Chulalongkorn Memorial Hospital, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "Amblyomma geoemydae is reported for the first time in this study. Its entire mitogenome is 14,780\u2009bp in length, contained 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and two non-coding regions. The phylogenetic analysis by Maximum-likelihood method show that A. geoemydae and the others of genus Amblyomma are in the same clade, indicating that A. geoemydae belongs to the genus Amblyomma.The complete mitochondrial genome of Amblyomma geoemydae (Ixodida: Ixodidae) is a reptile-associated tick species widely distributed the Southern part of Asia, ranging from South India to the Philippines, Australia and Japan are blood-feeding ectoparasites of terrestrial vertebrates . Species identification was conducted by professor Sun Yi based on morphological features. Primers were designed for polymerase chain reaction (PCR) amplification and sequencing on the basis of the mitogenome sequence of A. cajennense (GenBank accession no. NC_020333) , which contained 13 protein-coding genes , two rRNA genes, 22 tRNA genes, and two NCR. The arrangement of the A. geoemydae was identical with that of hard ticks , using Nuttalliellidae namaqua (NC_019663) as out group.The total length of A. geoemydae and the others of genus Amblyomma are in the same clade (A. geoemydae to the genus Amblyomma.The result show that the tree divided into two large branches: Prostriata and Metastriata. Phylogenetic analysis revealed that the me clade . This re"} +{"text": "In parallel with life-threatening major diseases such as cardiovascular events, cancers, and diabetes, chronic pain is another leading source of global people's sufferings and disabilities . Pain inIn this special issue, readers find eight articles, covering a wide spectrum of musculoskeletal pain. In detail, there are five articles focusing on the management of pain . An article by Y. Wang et al. addresses the recovery process for patients with lumbar disc herniation undergoing percutaneous endoscopic lumbar discectomy. One article presents the clinical outcome prediction for adolescents undergoing spinal fusion surgery. One article profoundly analyzes the state-of-the-art of available evidence regarding lateral epicondylitis by K. L. Ma et al. In terms of body regions, there are two articles focusing on extremities , three articles on the spine , one on the pelvis , and one on the head . One of the articles is not related to body parts, but describes the potentialities of the neuromodulation therapy .Collectively, this special issue presents emerging evidence for musculoskeletal pain in various aspects. In consideration of the high prevalence of pain, it deserves a great attention by the readers."} +{"text": "The integration of unmanned aerial vehicles (UAVs) with a cognitive radio (CR) technology can improve the spectrum utilization. However, UAV network services demand reliable and secure communications, along with energy efficiency to prolong battery life. We consider an energy harvesting UAV flying periodically in a circular track around a ground-mounted primary transmitter. The UAV, with limited-energy budget, harvests radio frequency energy and uses the primary spectrum band opportunistically. To obtain intuitive insight into the performance of energy-harvesting, and reliable and secure communications, the closed-form expressions of the residual energy, connection outage probability, and secrecy outage probability, respectively, are analytically derived. We construct the optimization problems of residual energy with reliable and secure communications, under scenarios without and with an eavesdropper, respectively, and the analytical solutions are obtained with the approximation of perfect sensing. The numerical simulations verify the analytical results and identify the requirements of length of sensing phase and transmit power for the maximum residual energy in both reliable and secure communication scenarios. Additionally, it is shown that the residual energy in secure communication is lower than that in reliable communication. Traditional wireless spectrum standards rely on the static spectrum allocation policies where a specific frequency band is assigned to the specific licensed users. Such a policy causes unbalanced spectrum utilization and degrades the spectral efficiency. Therefore, strict spectrum allocation is insufficient to meet the ever-growing demands of spectral resources for futuristic networks such as the internet of things (IoT), and 5G ,2. The fMoreover, signals in open and shared wireless medium are vulnerable to the eavesdropping, i.e., data interception by the illegitimate eavesdroppers. Traditionally, secure wireless data transmission requires cryptographic techniques at network layer. However, the information-security based on encryption and cryptographic techniques is not sufficient for the secure communications because advances in hardware design significantly increase the computational capabilities of the eavesdroppers. Different from the traditional cryptography algorithms, physical layer security (PLS) utilizes the transmission techniques and inherent properties of the wireless medium. The PLS is considered one of the potential solutions for the secure communication in wireless channels ,9. The POn the other hand, unmanned aerial vehicles (UAVs) have been used for various applications, such as monitoring, surveying, data transmission/communication, aerial remote sensing, product delivery, traffic control, and agriculture mapping . HoweverFurthermore, energy harvesting (EH) in wireless networks is the process of extracting energy from the surrounding environment, such as from solar, heat, wind, and radio frequency (RF) signals ,15. The We consider the energy management aspect of the energy harvesting CR-based UAV with limited-energy budget. The closed-form expressions of the total residual energy, connection outage probability, and secrecy outage probability are derived under a circular flight condition.We aim to extend the on-board battery life-time for UAV by maximizing the energy obtained through the EH and minimizing the transmission energy consumption. Thus, the optimal lengths of sensing phase and the transmit powers are obtained by solving the formulated optimization problems of maximum residual energy under the constraints of connection and secrecy outage probabilities with perfect sensing approximation.The analytical results are verified through the numerical simulations including imperfect sensing. Based on the results, we provide guidelines in designing an energy harvesting UAV-based CR system with the reliable and secure communications under scenarios without and with an eavesdropper, respectively.The main contributions of this paper are summarized as follows:The rest of the paper is organized as follows. In v with PT locating at the center. The altitude of EH-UAV relative to the PT is h, and the radius of circular track is given by r. The on-board power supply is responsible for the energy required for the flight operations, i.e., hovering and transition, with recharging possible only after the completion of each flight. The EH-UAV opportunistically exploits the primary-band, i.e., owned by PT and PR, in the absence or presence of E. The PT and PR communicate probabilistically during the flight of EH-UAV, which is divided into the sensing and transmission radians, i.e., sensing and transmission periods. In the sensing phase with a duration of t, the EH-UAV simultaneously harvests RF energy from the received signal and performs the spectrum sensing (SS) procedure for the opportunistic use of the primary band. The dynamic power splitting device splits the received primary signal into the two power fractions of The system model consists of an energy-constrained UAV as a secondary (cognitive) transmitter, a corresponding secondary receiver (SR), a ground-mounted primary transmitter (PT) and receiver (PR) pair, and an eavesdropper (E), as shown in g factor . In the RF power ,17. DuriRF power . In propectively ,20. An Onth samples of the signals received for the SS and EH, denoted by The simplest non-coherent energy detection method is considered for the spectrum sensing at the EH-UAV because it has low computational complexity and does not involve any complicated signal processing. The target signal, i.e., the signal from PT, is detected by comparing the measured signal energy with a pre-determined sensing threshold. It does not need any prior knowledge of the target signal . In the N is the total number of samples in the sensing phase. Let The decision metric, arameter ,20,21. TN, the decision statistic By using the central limit theorem, for large essed as (4)Pd=Q\u03f5ribution .Based on Equations and 5),,5), the essed as (6)Pfa=Qt and The sensing and transmission durations, i.e., Moreover, the spectrum sensing distance Given Equations and 8),,8), we cIn the sensing phase, the harvested energy by the EH-UAV is given aspower P\u03b4 . Hence, From Equations \u201312), th, th12), In the transmission phase, the harvested energy can be obtained asFrom Equations \u201316), th, th16), With Equations and 17)17), the The main objective of the EH-UAV integrated CR system is to use the primary spectrum for its opportunistic transmissions ,25,26. UThe connection outage probability is defined as the probability that the spectral efficiency (channel capacity) of the UAV-SR link (te (RS1) . Thus, tHere, For the evaluation of connection outage probability, the distribution of x and 0. Moreover, As a performance measure of PLS, the secrecy capacity Cs=Equation and Ce iThe secrecy outage probability is defined as the probability that the Equation can be rTo evaluate the secrecy outage probability, the distribution of SNRs for the UAV-SR and UAV-E links are required. Considering the distributions ained asPSec.Out=The total residual energy and two outage probabilities, including imperfect sensing, i.e., false alarm and detection probabilities, are too complicated to formulate and solve as an optimization problem, i.e., to maximize the total residual energy under outage constraints. For making the optimization problem tractable, the perfect spectrum sensing is assumed, i.e., Two maximization problems of In this subsection, a connection outage constraint is considered for a scenario without E. The optimization problem to maximize the residual energy with a connection outage constraint is formulated asThe optimization problem includes two variables: the length of the sensing phase in In addition, from quations and 44)P\u02dcS.Out\u2264\u03c6quations and 43)P\u02dcS.Out\u2264\u03c6Finally, Cognitive radio (CR) is a promising enabler communication technology to mitigate the spectrum scarcity and under-utilization issues in futuristic networks such as the IoT and 5G. The integration of UAVs in CR systems enhances the sensing performance. The wireless energy harvesting technique effectively alleviates the energy scarcity in the UAV-enabled wireless networks. Furthermore, most of the UAV based scenarios demand reliable and secure communications. In this paper, we considered that the UAV, with the limited energy-budget and a circular flight track (around a ground-mounted primary transmitter), harvests RF energy from the primary transmissions and uses the primary spectrum opportunistically. The closed-form analytical expressions for the residual energy, connection, and secrecy outage probabilities were derived to investigate the performances of energy-harvesting, reliable, and secure communications, in the absence and presence of an eavesdropper, respectively. The optimization problems were constructed by exploiting the trade-off between monotonic approximated functions, and the analytical solutions, i.e., the optimal lengths of sensing phase and transmit powers, are identified under two different scenarios for the UAV. The numerical simulations verified the proposed theoretical analysis, and demonstrated the impact of system parameters on the residual energy performance while ensuring the reliable and secure communication for UAV."} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-020-75996-5, published online 02 November 2020Correction to: The original version of this Article contained an error.The product name for the dicamba herbicide was included, however, due to issues relating to confidentiality, it has since been removed from the figures, the text, and tables. This information is available from the authors, and the data availability section has been updated to reflect this.These errors have now been corrected in the PDF and HTML versions of the Article."} +{"text": "Dr. Chen is one of the founding Editors of our Journal of Biomedical Science. Despite his extremely busy daily activities in clinics, teaching, research, and administration, he had served continuously as an associate editor until the very last day, when he bid his farewell to all of us. Over the past 25\u2009years, he witnessed the long and winding road of this Journal\u2019s growth and struggle - from impact factor (IF) zero to being enlisted by JCR (IF 0.99), to the most recent IF 5.762 in 2019. His death is truly a big loss to our Journal and to the entire medical and scientific community, in Taiwan and worldwide!Dr. Chen was born in the Ying-Ge Township, New Taipei City, Taiwan, in 1943. He graduated from National Taiwan University, College of Medicine, in 1968, and received medical residency training at NTUH in 1969\u20131973. Professor Sung Juei-low at NTUH is considered the father of liver disease research in Taiwan. In 1973, Dr. Chen chose to specialize in gastroenterology, and made the best decision in his career by joining Professor Sung\u2019s research team. He was involved in measuring the prevalence rate of hepatitis B virus (HBV) surface antigen (HBsAg) in liver disease patients. In 1975, Dr. Chen became an instructor at NTU. Arranged by Professor Sung, for four months in 1974\u20131975, he learned how to use new methods to detect HBsAg, and performed its serotyping, in the laboratory of Dr. Kusuya Nishioka at National Cancer Center Research Institute in Tokyo, Japan. Later in 1979\u20131980, he was a visiting scientist in Dr. Robert H. Purcell\u2019s laboratory at NIH in Bethesda, USA. In this one-year period, he was first exposed to molecular biology by studying HBV DNA integration in hepatomas.the Lancet (1983) the efficacies of hepatitis B immune globulin (HBIG) in blocking maternal-infant infection, as well as hepatitis B vaccine in its antibody production. HBV vaccine appeared to be safe for immunization of Taiwanese infants. Sung and Chen were then serving on a government consultation team. They advocated actively the vaccine idea to the society and Taiwan government, leading to a nationwide vaccination program for the newborn babies in Taiwan in 1984. Ms. Hsu-Mei Hsu, a card-carrying epidemiologist in the government sector, who happens to be Dr. Chen\u2019s wife, also played a key role in the entire planning and successful execution of this bold and unprecedented nationwide vaccination program in history! According to a classic paper published in 1997 in the New England Journal of Medicine (first authored by professor Mei-Hwei Chang and last authored by DS Chen), one decade after launching the vaccination program, the carrier rate in the vaccinated population dropped from 15% to less than 1%. Furthermore, the liver cancer incidence in children also decreased significantly. This is the first successful example of a human cancer vaccine!Professor Sung and his prot\u00e9g\u00e9 DS Chen demonstrated that the prevalence rate of HBsAg in Taiwan could be as high as 18%. They also noted that one common mode of virus transmission is the mother-to-infant transmission during delivery. Soon after his return from NIH to NTUH, Taiwan, in 1981, Chen and Sung would like to test if the HBV vaccine could be effective in reducing the incidence of viral hepatitis and liver diseases in Taiwan. The timing was indeed perfect, when Dr. Palmer Beasley, in close collaborations with Professor Chin-Yun Lee and several hospitals in Taipei, reported in In addition to hepatitis B, Dr. Chen and his colleagues also made significant contributions to the treatment of hepatitis C. For example, Drs. Ming-Yang Lai and Chen succeeded in treating hepatitis C patients with the combination of alpha-interferon and ribavirin. Back in the earlier days, this combination therapy was the best approach for treatment of hepatitis C.In Dr. Chen\u2019s illustrious career, he received numerous prestigious awards and recognitions. For example, in 1992, he was elected to be an Academician, Academia Sinica, Taiwan. In 2002, he received an honorary degree from Kaohsiung Medical University, the best medical school in Southern Taiwan. In 2005, he was elected as a member of the US National Academy of Sciences, a rare honor bestowed to few foreign associates. In 2006, he received the Trieste Science Prize from Third World Academy of Sciences (TWAS). In 2007, he was the recipient of the Presidential Award in Taiwan. In 2009, EASL International Recognition Award. In 2010, Japan Nikkei Asia Prize. In 2011, he was honored with a Distinguished Clinician Educator/Mentor Award from AASLD (American Association for the Study of Liver Diseases). In 2014, Fellow of AASLD. In 2016, EASL Hall of Fame. In 2018, the Blumberg Award from the Hepatitis B Foundation, USA. Dr. Baruch Blumberg won the Nobel Prize for his discovery of hepatitis B virus. As of January, 2019, Dr. Chen published a total of 750 original articles, 44 book chapters, and edited 3 books.Dr. Chen is also known as a talented administrator. For example, in 2001\u20132007, he served as the Dean of National Taiwan University, College of Medicine. For the sake of a more concise obituary, we will not go into details of his many important positions in the government, university, hospital, and professional societies. In appreciation of his many important contributions to clinical medicine and the national healthcare policy, Dr. Chen was honored with a 2nd-ranked Jin-Hsing Medal from the current Taiwan President Tsai Ing-wen in 2018.Dr. Chen is known to be a very hard-working person. He likes to compare himself to the domesticated water buffalo in Taiwan, which are very common in the farmers\u2019 rice field during his childhood. In his office, he collected many paintings, sculptures, and photos of water buffalo. It is clear that he is a believer in the virtue of hard-working quietly without complaints.Although Dr. Chen officially retired from NTU in 2013, he continued to be active in liver research and teaching young doctors in the NTUH hospital. Dr. Chen is survived by his wife Hsu-Mei Hsu \u2013 an epidemiologist; his son Chih-Heng (Henry) Chen - in the finance field, and his daughter Yun-Ru (Ruby) Chen \u2013 a protein chemist in Academia Sinica, Taiwan. Last, but not the least, Dr. Chen is also survived by many of his students and trainees, who have been active and successful in liver disease research. There is no doubt that the legend of Dr. Ding-Shinn Chen and his Taiwan buffalo spirit will continue to live on in generations to come!A public memorial service for Dr. Ding-Shinn Chen will be held at National Taiwan University, College of Medicine, Lecture Hall, on August 16, 2020."} +{"text": "Self-regulation is a multidimensional construct that is positively related to academic achievement, such as successful mathematics performance. However, this relation of self-regulation and mathematics performance has mainly been investigated in Western countries with similar cultural contexts, although self-regulation is assumed to be context-sensitive. Therefore, the present study investigated the relation of self-regulation and mathematics performance across two different countries (Germany vs. Iran) in college students. The relation of self-regulation and mathematics performance was expected to be weaker in students of math-related fields, such as Engineering/Informatics, as they are assumed to need less self-regulation to solve the mathematics problems than students of less math-related fields, such as Human Sciences. In total, 122 undergraduate students of Human Sciences or Engineering/Informatics participated in this study. We measured self-regulation with the Brief Self-Control Scale and math Self-regulation is defined as the ability to control one\u2019s thoughts, behaviors, or emotions, and enables individuals to adapt their behaviors in accordance with the demands of a situation e.g., . It inclPrevious studies have indicated that self-regulation contributes to mathematical performance by suppressing distracting thoughts or information whilst mathematics problems are solved e.g., , and thrHowever, the relation of self-regulation and mathematics performance might vary across different contexts. Recent studies demonstrated that self-regulation is a context-specific construct e.g., , suggestindependent and interdependent contexts, which can influence self-regulation. Independent contexts focus on autonomy and individual goals, whereas interdependent contexts are associated with being in harmony with the group and the community goals. Accordingly, self-regulation processes in an independent context are directed toward influencing the environment and other people in line with an individual\u2019s goals, while in interdependent contexts they focus on adjusting one\u2019s behavior to the expectations of others to maintain fit with the group and 62 Iranian undergraduate students. The German participants were recruited from the University of T\u00fcbingen in south Germany and Iranian participants were from the University of Tehran, Iran. All participants were native speakers with no immigration backgrounds. The entire data of the participants were analyzed anonymized . Detailed characteristics of both German and Iranian students are depicted in Participants were 60 GermanBackground characteristics, consisting of field of study, math score in the University entrance exam, math self-concept, expectancy of success, and demographics of the participants were collected with a background questionnaire. The questions of the background questionnaire, except the questions of math self-concept, were developed by the authors. Math self-concept was assessed by four questions based on the SDQ (Self Description Questionnaire) III .Participants\u2019 self-regulation was assessed by using self-reports. Participants were asked to fill out the Brief Self-Control Scale BSCS; . The Gercompletely true) to 5 (completely untrue). Nine items were reverse-coded and the total score was the sum of the responses of all items, with higher sum scores representing more self-regulation. In the present study, the questionnaire showed sufficient internal consistency .The BSCS consists of 13 items targeting thought control, impulsive response control, action persistence, and action monitoring . The response format was a 5-point Likert-type scale ranging from 1 (L and A on a German keyboard) for correct and incorrect solutions, respectively. The response keys were counterbalanced across participants. Except for practice trials, all trials were presented without feedback.Mathematics performance was assessed by using the complex multiplication test, consisting of 48 complex multiplication problems. The complex multiplication problems entailed one-digit times two-digit problems with two-digit solutions and lower error rates (ERs). Multiplication RTs of the participants were defined by the time intervals between the presentation of the multiplication problems on the screen and the responses of the participants, measured by pressing the keys of the computer keyboard. Only RTs of correct responses were considered in the analyses. Moreover, RTs shorter than 200 ms were excluded, and subsequently RTs which were more or less than \u00b1 3 SD around the individual mean were excluded continually until no more outliers remained with self-regulation as predictor and mean multiplication RTs as outcome variable. In the second step, to compare the relation of self-regulation and mathematics performance between German and Iranian students, the linear regression analysis was calculated with self-regulation, country (dummy coded), and the interaction between self-regulation and country as predictors and mean multiplication RTs as the outcome variable.The second hypothesis of the present research was that the relation of self-regulation and mathematics performance is weaker in students of Engineering/Informatics. In the first step, four separate linear regression analyses were conducted for each subsample field of study with self-regulation as predictor and mean multiplication RTs as outcome variable. In the second step, to compare the relation of self-regulation and mathematics performance in students of Human Sciences and Engineering/Informatics, the interaction between self-regulation and field of study was tested in a multiple linear regression analysis with self-regulation, field of study (dummy coded), and the interaction between self-regulation and field of study as predictors and mean multiplication RTs as the outcome variable. All continuous variables were standardized and the level of significance was set to \u03b1 < 0.05 for all analyses.p-values < 0.05), Mann\u2013Whitney U test, and for normally distributed variables t-test and Fisher\u2019s Exact Test were used.Descriptive and test statistics for the background characteristics and the study measurements of German and Iranian students are presented in U = 2.40, p = 0.005, gender, p = 0.011, Fisher\u2019s Exact Test, and expectancy of success, U = 1.25, p = 0.001. Although German and Iranian students significantly differed in age and gender = \u22124.46, p < 0.001, d = 0.81, and Iranian students made more errors, U = 2.62, p < 0.001. However, German and Iranian students did not differ in math self-concept, U = 1.92, p = 0.738, and self-regulation, t(120) = 1.21, p = 0.229, d = 0.22.Additionally, German and Iranian students did differ in their multiplication performance: German students were slower, b = \u22120.25, t = \u22121.10, p = 0.051; see b = \u22120.09, t = \u22120.72, p = 0.473; see b = \u22120.09, t = \u22120.53, p = 0.599; see VIF = 2.75; country, tolerance = 0.99, VIF = 1.01; self-regulation \u00d7 country, tolerance = 0.37, VIF = 2.73), independent errors , and non-zero variances and contained no outliers .Regression analysis revealed that self-regulation did not predict multiplication RT neither in German = 5.12, p = 0.029] and Iranian students of Human Sciences, but not in German and Iranian students of Engineering/Informatics , independent errors , and non-zero variances and contained no outliers .Moreover, the non-significant interaction indicates that the relation of self-regulation and mathematics performance did not significantly differ between students of Human Sciences and Engineering/Informatics . As the second hypothesis, we expected that the relation of self-regulation and mathematics performance was weaker in students of Engineering/Informatics as compared to students of Human Sciences. Contradictory to our first hypothesis, the relation of self-regulation and mathematics performance did not differ between German and Iranian college students: self-regulation did not predict multiplication RT neither in German nor Iranian students. Moreover, inconsistent with our second hypothesis, the results showed that the relation of self-regulation and mathematics performance did not differ significantly between students studying less math-related fields and students of math-related fields in the whole sample. However, partially in line with our second hypothesis, when the field of study was considered within the countries, self-regulation predicted multiplication RT in those students studying Human Sciences but not in students of Engineering/Informatics within each country. Thus, although the main effect of field of study was not observed regardless of country, the relation of self-regulation and mathematics performance seemed to be descriptively weaker in students of Engineering/Informatics than Human Sciences within each country. This might be because the complex multiplication test within each country seemed to be less difficult for the students of Engineering/Informatics compared to the students of Human Sciences, therefore, these students might need less self-regulation to solve the problems. The complex multiplication test seemed to be less difficult for the students of Engineering/Informatics as they performed better than students of Human Sciences in general see . HoweverTaken together, the results showed that the relation of self-regulation and mathematics performance did not differ between German and Iranian college students. Furthermore, we observed this similarity not only in the context of country but also in the context of field of study, which is further supported by the fact that when only the students of Human Sciences are compared, the association between self-regulation and mathematics is similar in both countries . This fiHowever, our finding is in contrast with previous studies, connecting the academic achievement gap between students from different countries to the effect of cultural context on self-regulation. For instance, in a longitudinal study by Altogether, cultural context did not seem to play a dominant role in moderating the relation between self-regulation and math performance in the present study. However, with regard to the confounding effect of field of study within each country on the predictive validity of self-regulation, careful sample selection considering field of study of students is recommended for future research examining the relation of self-regulation and mathematics performance.The current research has some limitations worth noting. First, there might be structural and cultural variations in educational systems such as different grading systems or teachers\u2019 expectations, as well as academic motivation of students within and between nations that may differentially influence self-regulation and its relation with academic performance. Therefore, we view this study only as a starting point for investigating the impact of independent and interdependent cultures on the relation of self-regulation and math performance. Future studies conducted in other independent or interdependent cultures should clarify whether the observed results are really due to this cultural difference or to other educational or cultural differences, which are particular to the specific countries studied here. Second, German students of Human Sciences were offered different reimbursement than other participants since the study in which they participated, was part of a larger project consisting of 4-h experiment. Hence, we acknowledge that different incentives in German students of Human Sciences in comparison to other participants might generate participation bias and account partially for the findings of the current study. Third, self-regulation consists of several components such as cognitive, behavioral, and emotional aspects that are differentially related to mathematics performance and their effects should be investigated individually in the future research. Forth limitation is the small sample size of the present study that may preclude a definitive statement for the present study. The last, but not least, important limitation is construct validity in the present study, as our research measurement for assessing self-regulation was designed and validated for Western countries. The problem is that in self-reports, participants of one cultural context may interpret the words differently and compare themselves with different standards than those in another cultural context e.g., . In our In conclusion, our findings show that the relation of self-regulation and mathematics performance is similar in German and Iranian college students. In addition, the effect of field of study on the relation of self-regulation and mathematics performance was highlighted in the present study. Self-regulation did not predict mathematics performance in German and Iranian students, however, when the effect of field of study was taken into account, self-regulation predicted mathematics performance in students of less math-related fields of study within each country. It is important to note that while the single analysis produced differential results, a direct comparison of the different fields of studies was non-significant \u2013 therefore, we have interpreted these results with great care. Nevertheless, since the relation between self-regulation and mathematics performance within each country, was significant only for less math-related fields of study, we suggest that the possible confounding effect of field of study should be considered in studies when the relation of self-regulation and mathematics performance is examined.The datasets generated for this study are available on request to the corresponding author.Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. The patients/participants provided their written informed consent to participate in this study.PN and CG designed and performed the research. PN and JK analyzed the data. PN, JK, CG, and H-CN wrote the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Apelin, an endogenous neuropeptide, has been identified as the cognate ligand for the G-protein-coupled receptor APJ. Apelin, APJ messenger RNA, and protein are widely expressed in the central nervous system and peripheral tissues of humans and animals. The apelin/APJ system has been implicated in diverse physiological and pathological processes. The present article reviews the progress of the latest research investigating the apelin/APJ system in pain, depression, anxiety, memory, epilepsy, neuroprotection, stroke, and brain injury and protection, and highlights its promising potential as a therapeutic target for treatment of psychosis and neuropathy. Apelin (APLN), an endogenous ligand of the APJ receptor (APLNR), was first isolated from bovine stomach tissue . The preelabela was comprised of three exons on chromosome 4. The elabela encodes a conserved 54-amino acids protein, containing an N-terminal signal-peptide and a mature 32- amino acids peptide, named Elabela signaling (i/o stimulating mitogen-activated protein kinase (MAPK) cascade via protein kinase C (PKC) or intrathecal administration of apelin-13 resulted in a marked antinociception in the mouse tail-flick test . In the ignaling and coup C (PKC) .Recently, Turtay et\u00a0al. reported that intraperitoneal (i.p.) injection of apelin-13 (100 \u00b5g/kg) exerted an analgesic effect in both the hot-plate and the tail-flick tests in rats, and that antinociception was reduced by ondansetron . Chronicapln and aplnr mRNA, and APLN and APLNR protein than vehicle control, and apelin-13 exerted no effect on the neuropathic nociceptive response (The apelin/APJ system plays a role in chronic (neuropathic) and acute pain. Chronic i.t. injection of Pyr-apelin-13 (1 and 5 \u00b5g/rat) attenuated neuropathic pain and reduced caspase-3 levels in rat spinal cord tissues . The spiresponse . Howeverresponse .apln/aplnr mRNA and APLN/APLNR protein expression in the spinal cord, suggesting that EA stimulation could inhibit inflammatory pain, in part, by restoring apln/aplnr mRNA and APLN/APLNR protein decreased pain threshold in the rat tail-flick test . In the protein .These inconsistent results of the apelin on pain regulation are dif\ufb01cult to explain. It may be due to the different kind of animal model of pain, doses, animal species, administration routes, time of injection, forms of apelin, etc. The main molecular mechanism of apelin/APJ on pain was related to opioid receptor, GABA receptor, and ERK pathway. The effect of apelin/APJ in pain animal models had been extensively studied. However, the roles in primary afferent inputs, pain modulation at the spinal level, and plasticity after nerve injury or inflammation remain unclear. The apelin/APJ systems may be developed as novel analgesics.aplnr mRNA has been found in the amygdala, hypothalamus, Ammon's horn, and the dentate gyrus axis has been observed in depressed patients . Persist2 serotonergic receptors exhibited an anxiolytic effect in the elevated plus maze, and the antianxiety of apelin-13 was mediated by \u03b1-adrenergic, \u03b2-adrenergic, dopaminergic, and 5-HTeceptors . Chroniceceptors . This efeceptors . Additioeceptors . Apelin-The different effects of apelin on depression, however, may be due to different injection methods and/or different animal species. Moreover, It was reported that the forced swim test does not re\ufb02ect depression , which mapln/aplnr mRNA and APLN/APLNR protein have been found in the hippocampus, amygdala, and cerebral cortex prevented neuronal apoptosis by suppressing the generation of reactive oxygen species, cytochrome c release, mitochondria membrane depolarization, and caspase-3 activity . Intravieceptors .Ischemic stroke is a common neurological disease, and generally leads to brain damage and neuronal cell death . Chen etaplnr variant and ischemic stroke. The rs9943582 variant of aplnr was associated with a significantly higher risk for brain infarction in the Japanese population , and the increase of AQP4 caused by apelin-13 was mediated through the PI3K/Akt and ERK pathways , the Plan for Scientific Innovation Talent of Henan Province and Henan Provincial Natural Science Foundation (Grant No. 182300410323 and 182300410316), the Program for Science & Technology Innovation Talents in Universities of Henan Province to W-DC; the National Natural Science Foundation of China (Grant No. 81600974 and No. 81971280), the Key Science and Technology Program of Henan Province in China (Grant No. 192102310080), the Key Scientific Research Program for Universities of Henan Province in China (Grant No. 17A310003), the Fundamental Research Funds of Henan University (Grant No. yqpy20170040), the Key Science and Technology Program of Kaifeng City in China (Grant Nos. 1803034 and 1903019), and the Scientific Research Foundation of Henan University (Grant No. 2015YBZR050) to S-YL; the National Natural Science Foundation of China , the Fundamental Research Funds for the Central Universities and Research Projects on Biomedical Transformation of China-Japan Friendship Hospital (Grant No. PYBZ1803), and the Fundamental Research Funds for the Central Universities (Grant Nos. PYBZ1706) to Y-DW.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The separation and sorting of micro- and nano-sized particles is an important step in chemical, biological, and medical analyses. In the past two decades, micro- and nanofluidic platforms have been increasingly applied for the separation, fractionation, sorting, and purification of all classes of particles based on their physical and chemical properties because of their advantages of minimal consumption of sample and reagent, ease of use, and enabling of the integration of multicomponent for comprehensive analysis. The separation techniques using micro- and nanofluidic devices are classified into passive methods using geometries and hydrodynamic effects at micro/nanoscale, and active methods using external fields such as electric, magnetic, optical, and acoustic forces.This Special Issue collects some state-of-the-art developments in active and passive microfluidic separation, isolation, and manipulation for a wide range of particles. In this Special Issue, 11 research papers, and two review articles are published. Five papers ,3,4,5 an1) Passive microfluidic technique: Bogseth et al. proposed a co-flow inertial microfluidic device that is tunable in multiple ways for adaptation to different application requirements Passive . Jiao et(2) Active microfluidic technique: Krishna et al. presented an experimentally validated mathematical model of a microfluidic device with nozzle-shaped electrode configuration for dielectrophoretic 3D-focusing of particles . They inWe would like to thank all authors for submitting their papers to this Special Issue. We would also like to acknowledge all the reviewers for dedicating their time and timely reviews to improve the quality of this Special Issue."} +{"text": "On the other hand, in the theory of alternating automata, conjunction and non-determinism are core aspects. Alternating automata have not been considered in the context of inputs and outputs, making them less suitable for modelling software interfaces. In this paper, we combine the two modelling paradigms to define alternating interface automata (AIA). We equip these automata with an observational, trace-based semantics, and define testers, to establish correctness of black-box interfaces with respect to an AIA specification.To model real-world software systems, modelling paradigms should support a form of compositionality. In interface theory and model-based testing with inputs and outputs,"} +{"text": "Metcalfa pruinosa inferred the origin of the species using a fragment of mitochondrial COI sequences. However, the low variability of the sequences limited further scrutinized inference on the invasion dynamics. In this study, we sequenced a fragment of the COI gene from 536 individuals of the species and combined the sequence data with the available GenBank data, totaling 830 individuals. These data indicated that the North-West region is a point of entry in addition to the South-East region, the presumed sole point of entry to Korea. Furthermore, it suggested that North-West entry involves the M. pruinosa originating from the USA. In an effort to find further variable regions in the mitochondrial genome, one region provided substantially increased variability compared to the fragment of the COI. The concatenated sequences of COI and the newly obtained variable region, which were used to infer the expansion pattern in Korea, indicated that the main highway, running obliquely between the North-West and South-East regions, appears to be responsible for the current population genetic structure of M. pruinosa in Korea, facilitating gene flow through this highway traffic.After the invasion of Korea in 2005, the first lines of studies on Metcalfa pruinosa (Hemiptera: Flatidae), which is an invasive species, is widespread in Korea. We sequenced a fragment of the COI from 536 individuals collected mainly in Korea and the European countries and combined these sequence data with the public data, totaling 830 individuals worldwide. The identification of one shared haplotype only between Korea and the USA, the presence of this haplotype only in the North-West region of Korea, and the highest haplotype diversity in this region suggested that the North-West region is another point of entry in addition to the South-East region, which is the presumed sole point of entry to Korea. Furthermore, it suggested that North-West entry involves the M. pruinosa originating from the USA. In an effort to find further variable regions in the mitochondrial genome, one region provided substantially increased variability compared to that of the fragment of COI. FST estimation, PCoA, and BAPS analysis, using the concatenated sequences of COI and the newly detected variable region to infer the expansion pattern in Korea, indicates that the main highway, running obliquely between the North-West and South-East regions, appears to be responsible for the current population genetic structure of M. pruinosa in Korea, facilitating gene flow through this highway traffic.The flatid planthopper, Metcalfa pruinosa (Say) (Hemiptera: Flatidae), which is native to North America, including Mexico and Cuba . A preliminary test using a few geographic samples of M. pruinosa has shown that one region in the A + T-rich region (Region 5) has higher variability than that of the DNA barcoding region. The primers designed from these mitogenome sequences , number of haplotypes , maximum sequence divergence , number of populations with two or more haplotypes , and haplotype diversity . When thAmong the 20 haplotypes, MPRB01 and MPRB02 each accounted for 48.25% and 29.83% , respectively . In the Among the 17 haplotypes found in Korea, MPBR01, MPBR02, and MPBR16 were the most widely distributed in 14, 11, and 6 localities, respectively; others were found in 1\u20133 localities . HaplotyH (max = 1.0) and \u03c0 per country, ranging from 0.6389 (Spain) to 0.7692 (France) in H and 0.000924 (Spain) to 0.004968 (France) in \u03c0, indicating that Spain was the lowest and France was the highest, although there was no substantial difference in both H and \u03c0 among countries when standard errors were considered to 0.7118 (North-West), and \u03c0 from 0.000933 (South-West) to 0.002217 (North-West), with substantially higher H in the North-West region and a substantially lower H in the South-West region, but the differences observed in \u03c0 were not substantial. This regional subdivision further indicates that the H in the North-West region is the second highest, next to that of France , North-East , South-West , and South-East regions to 0.39379 (between Spain and France), with a range of mN from 288.6 to 0.770 (between Spain and France) (p < 0.001) was observed in all country pairs, except for the comparisons between Italy and France. When the Korean localities were divided into the four regions, as we did for the diversity analysis, a significant FST (p < 0.001) was detected in four comparisons, but not in the comparisons between the North-East and South-West regions and between the North-West and South-East regions than between those located adjacent to each other (between North and South regions).The France) a. A sign regions b. BetweeK = 8 showed that all eight haplogroups were found in Korea, three in Italy, two in Spain, and five in France from 294 individuals, totaling 830 individuals. However, we did not obtain any haplotypes from Korea in addition to those reported by Park et al. [To obtain further detailed inference on the origin of the Korean populations compared to previous studies ,17, we ek et al. , MPH03 iM. pruinosa was detected in Korea [M. pruinosa accompanying MPH03, most likely from the USA, has been introduced through the North-West locality to Korea independently from Gimhae. Indeed, it is highly unlikely that a derived population can show evidence of a haplotype that is not found at the original point of entry, particularly considering the diffusional nature of the dispersal pattern of mitochondrial haplotypes [H and \u03c0 , which is located in the South-East region close to Gimhae, is the largest trading port in the country and handles an enormous quantity of agricultural products transported from diverse countries, including Europe and the USA. In addition, there is an international airport in Gimhae, but airplanes arriving there are only from a few Asian countries (https://airport.co.kr). In the North-West region, the Incheon (https://icpa.or.kr) and Pyeongtaek-Dangjin ports (https://gppc.or.kr) are the fourth and fifth largest ports, respectively. Similar to the Busan port, these ports handle similar quantities of agricultural produce and hardwood and cargo ships from several countries including the USA and Europe. In addition, the largest international airport is located in the northwest region of Korea. Moreover, nationwide monitoring results [The Busan Port in Korea .In contrast to the COI data , and theCOI data consisteCOI data . SimilarM. pruinosa have been proposed [Previously, two patterns of dispersal responsible for the domestic spread of proposed ,37. One proposed ,39,40,41COI data did not allow the detailed investigation of the expansion dynamics of the Korean populations, the concatenation of the newly developed Region 5 to the DNA barcoding region was useful to infer expansion dynamics in Korea, and this approach allowed further thorough tracing of domestic expansion in Korea. The FST estimates between pairs of regions using the concatenated sequences revealed an interesting pattern. It showed a higher mN only between the North-West and South-East regions and between the North-East and South-West regions, providing non-significant FST, even at a p < 0.05 level and between the North-West and South-East regions (red) (Although the worldwide 05 level . Considens (red) . PCoA anns (red) .FST data may be explained by two major independent introductions in the North-West region and subsequent dispersal through highway road traffic . In an additional major introduction to the North-West region at a later time point, M. pruinosa bearing a higher frequency of MPBR02, which is the dominant haplotype currently in both the North-West (49.47%) and South-East (66.67%) regions, may have spread by these two patterns of dispersal. The Seoul\u2013Busan highway that directly connects the North-West and South-East regions, may have played a major role in ensuring longer dispersal (http://www.ex.co.kr/) as it connects Seoul, which is the capital of Korea and located in the North-West region, to Busan, which is the second largest city and located in the South-East region. The highway roughly bisects Korea diagonally into West and East regions (M. pruinosa entering through the South-East region is not obvious in the current population genetic structure, along with the origin. It is likely that earlier entry of M. pruinosa through the South-East region played a certain role in its spread to the neighboring localities, which were previously unoccupied, but our data do not show a definite role of M. pruinosa entering through this region.The current traffic . Considet region ,8,14, in regions . The detcalities , the earispersal . This 41 regions . Because regions . TemporaCOI data suggests that the invasion of Korea by M. pruinosa occurred in two independent regions, the South-East and North-West, both of which harbor major ports and airports that import diverse agricultural produce. Introduction into the North-West region demonstrates that M. pruinosa likely originated from the USA because an identical haplotype was detected in Korea and the USA. However, the origin of entry to the South-East invasion is inconclusive, and further studies on populations from additional regions, including North America, are required. The concatenated sequences of the DNA barcoding region and Region 5 allowed inference of the expansion pattern within Korea, indicating that the current population genetic structure can be explained by two independent introductions into the North-West region. These introductions involved varying frequencies of the two dominant haplotypes at each time, longer dispersal facilitated via the Seoul\u2013Busan highway, and resultant genetic connections only between diagonally located regions, such as between the North-West and South-East regions. As further samples from the introduced and native populations become available, improved inference regarding the origin and expansion pattern of M. pruinosa will be possible. This will be crucial for the prevention of additional invasions and quarantining of similar invasive species. This is essential considering the global biological invasion, despite the time that has elapsed since the first record of M. pruinosa in Korea.The worldwide"} +{"text": "Micromachines Young Investigator Award to:After an extensive voting period, we are proud to present the winner of the Junsuk Rho is currently a Mu-Eun-Jae endowed Chair Associate Professor with a joint appointment in the Department of Mechanical Engineering and the Department of Chemical Engineering at Pohang University of Science and Technology (POSTECH), Korea. Before joining POSTECH, he received his B.S. (2007) and M.S. (2008) degrees in Mechanical Engineering at Seoul National University, Korea, and the University of Illinois, Urbana-Champaign, respectively. After receiving his Ph.D. (2013) in Mechanical Engineering and Nanoscale Science and Engineering from the University of California, Berkeley, Junsuk Rho worked as a postdoctoral fellow in the Materials Sciences Division at Lawrence Berkeley National Laboratory and Ugo Fano Fellow in the Nanoscience and Technology Division at Argonne National Laboratory. His research is focused on developing novel nanophotonic materials and devices based on fundamental physics and experimental studies of deep sub-wavelength light\u2013matter interaction. Dr. Rho has published approximately 150 high-impact peer-reviewed journal papers and has also presented keynote and invited talks more than 250 times at world-leading institutes and international conferences/workshops. He also has 4 US patents and 26 Korean patents.Micromachines Editorial Office staff and award evaluation committee, I congratulate Dr. Junsuk Rho on his excellent performance and wish him all the best for his future career.On behalf of the This is Junsuk Rho, who is currently a Mu-Eun-Jae endowed chair Associate Professor with a joint appointment in the Department of Mechanical Engineering and the Department of Chemical Engineering at Pohang University of Science and Technology (POSTECH), Korea.My research is focused on developing novel nanophotonic materials and devices based on fundamental physics and experimental studies of deep sub-wavelength light-matter interaction. Specifically, I am working on metamaterials, plasmonics, photonic crystals, topological photonics. Recently, the wavelength regime has expanded to longer wavelengths such as acoustic and elastic frequencies. I saw the news that invisibility cloak may be realized by metamaterials. That's the reason why I want to do this for my Ph.D. study. Such metamaterials will be able to realize invisibility cloaks, super-resolution imaging, acoustic black hole, hologram display and so on.Quantum Optics, Flexible Opto-Electronics, Photonic Chips for diagnostics, Single-digit-nanometer fabrication to overcome the current semiconductor industry.Everyday... This is related to #5. Have patience, try the research until it comes through without so much depressed and frustrated.Patience, Positive mind for failure , Catching up the trends. Open access journals allow more readers to read the papers, which is very good to share the result with the communities. However, with the limited funding , it sometimes becomes a burden. So, some kinds of supports for junior researchers are necessary. In terms of this, MDPI does a very good job to provide financial support and waive of the fee for the junior researchers."} +{"text": "Scientific Reports 10.1038/s41598-018-23954-7, published online 24 April 2018Correction to: In the original version of this Article, Mainak Dutta was incorrectly affiliated with \u2018Department of Biotechnology, Birla Institute of Technology Pilani (Dubai Campus), Dubai, United Arab Emirates\u2019. The correct affiliation is listed below.Department of Biotechnology, Birla Institute of Technology and Science, Pilani (Dubai Campus), Dubai, United Arab Emirates.This error has now been corrected in the PDF and HTML versions of the Article, as well as in the Supplementary Information file."} +{"text": "More than 30 million adults are released from incarceration globally each year. Many experience complex physical and mental health problems, and are at markedly increased risk of preventable mortality. Despite this, evidence regarding the global epidemiology of mortality following release from incarceration is insufficient to inform the development of targeted, evidence-based responses. Many previous studies have suffered from inadequate power and poor precision, and even large studies have limited capacity to disaggregate data by specific causes of death, sub-populations or time since release to answer questions of clinical and public health relevance.To comprehensively document the incidence, timing, causes and risk factors for mortality in adults released from prison.We created the Mortality After Release from Incarceration Consortium (MARIC), a multi-disciplinary collaboration representing 29 cohorts of adults who have experienced incarceration from 11 countries. Findings across cohorts will be analysed using a two-step, individual participant data meta-analysis methodology.The combined sample includes 1,337,993 individuals , with 75,795 deaths recorded over 9,191,393 person-years of follow-up.The consortium represents an important advancement in the field, bringing international attention to this problem. It will provide internationally relevant evidence to guide policymakers and clinicians in reducing preventable deaths in this marginalized population.Mortality; incarceration; prison; release; individual participant data meta-analysis; consortium; cohort. Each year more than 30 million people are released from incarceration globally and thisThere is an established literature documenting an elevated risk of premature death in the first four weeks after release from incarceration -16. HoweFew attempts to date have been made to synthesise the literature examining mortality after release from incarceration. Kinner et al. conducteTo address the knowledge gaps identified above, we have created the international Mortality After Release from Incarceration Consortium (MARIC). This aim of this paper is to describe the composition of the Consortium, its data, objectives and research methodology. The Consortium is an international, multi-disciplinary and multi-organisational collaboration of researchers, clinicians and policymakers and represents the largest coordinated effort to date worldwide to examine mortality in adults who have experienced incarceration. The Consortium is led by the University of Melbourne in Australia and is funded by Australia\u2019s National Health and Medical Research Council . It is an open Consortium and welcomes new collaboration proposals from academics, policymakers, clinicians, and service providers worldwide. The Consortium\u2019s current dataset is comprised of 29 cohorts of adults who have experienced incarceration from 11 countries: Australia, Canada, French Guiana (France), Indonesia, Malaysia, the Netherlands, Norway, Scotland, Sweden, Taiwan, and the USA examine specific (including rare) causes of death, and b) conduct meta-regression analyses to consider findings according to key demographic, policy-based and country-level variables, elucidating country-specific structural factors contributing to the observed heterogeneity in mortality estimates. Finally, due to the multi-disciplinary nature of the Consortium, interpretation of findings will also benefit from expert knowledge and experience across a wide spectrum of health and criminal justice settings.The main outcome of the Consortium is mortality after release from incarceration. The aims of the Consortium are to: 1) comprehensively establish the incidence and timing of all-cause and cause-specific mortality in adults following release from incarceration internationally; 2) identify risk factors for all-cause and cause-specific mortality following release from incarceration; and 3) examine how risk differs across settings, time, and specific sub-populations. The specific causes of mortality we will examine are:Non-communicable diseases ;Alcohol and other drug-related ;Suicide ;Infectious diseases ; andInjuries other than self-inflicted injuries and poisoning .Data analysis for the Consortium is structured around the aims outlined above and will involve a series of two-step, individual participant data meta-analyses (IPDM-A) see Figu. In the Strengths of our consortium include its large sample size, its multi-national composition, and its use of ICD codes to assign causes of death. Our Consortium has some limitations. First, 25 of the 29 cohorts (86%) come from high-income countries, with a geographical distribution concentrated in North America (n=12), Australia (n=8), and Western Europe (n=4). While this also represents an advantage, due to the broad similarities of the criminal justice systems in these regions, further data on the health of people who experience incarceration in low- and middle-income countries are urgently needed . To thisThe disproportionate rates of premature mortality experienced by adults released from incarceration represenImportantly, incarceration itself is a high-risk event for morbidity and mortality outcomes , 50 and The MARIC Consortium represents an opportunity to substantially improve the evidence base regarding mortality in adults released from incarceration and produce targeted, globally relevant evidence on the epidemiology of mortality in this population. The overarching aim of the Consortium is to substantially increase the accuracy, precision, clinical relevance and translational impact of research on mortality in adults following release from incarceration across countries. Findings and recommendations from the Consortium will lay the foundation for policy reform, targeted clinical intervention, and rigorous evaluation of scalable interventions that have the potential to reduce the unnecessary wastage of lives after release from incarceration internationally. https://mspgh.unimelb.edu.au/research-groups/centre-for-health-equity/justice-health-unit/mortality-after-release-from-incarceration-consortium-maric-study. Specific inquiries, including collaboration proposals, can be directed to the Consortium\u2019s Chief Investigator, Dr. Rohan Borschmann (rohan.borschmann@unimelb.edu.au). The Mortality After Release from Incarceration Consortium includes all authors listed above, in addition to Trudi Cooper, Neil Drew, Lisa Duffy, Michael Farrell, Cath Ferguson, Natalie Gately, Natasa Gisev, Ann-Claire Larsen, Jo Kimber, Richard Mattick, Paul Nieuwbeerta, Moira Sim, Di Twigg, and Jacqui Whale.Further information about the MARIC Consortium is located at: The MARIC study is funded by Australia\u2019s National Health and Medical Research Council . The authors wish to acknowledge the assistance of the Western Australian Department of Justice in the conduct of the research, along with all organisations, government departments, and other entities worldwide that have contributed to this research. The material published herein cannot be considered as either endorsed by the Western Australian Department of Justice or an expression of the policies or view of the Department. The opinions generated in this study do not represent the British Columbia Ministry of Health or any data stewards. Any errors of omission or commission are the responsibility of the research team.RB, SK, MS, JP, SL, DP, DR, EO, and LM obtained funding for the Consortium. RB produced the first draft of the manuscript. HT, JY and CK produced the tables and figures. All authors contributed to subsequent iterations of the manuscript and approved the final manuscript prior to submission.We did not involve patients or the public in our work.Ethics approval was granted for all 29 individual cohorts in the consortium, and no further approval was required for the broader collaborative study.TablesSupplementary File"} +{"text": "Cell Death and DiseaseCorrection to: 10.1038/s41419-020-02906-y published online 18 August 2020The original version of this article contained errors in the author affiliations.Author Xiao Li was incorrectly associated with Department of Obstetrics and Gynaecology, Shanghai Sixth People\u2019s Hospital, Shanghai Jiaotong University, Shanghai 200233, China. The correct affiliation is Shanghai Municipal Key Clinical Speciality, Shanghai 20030, China.This has now been corrected in both the PDF and HTML versions of the article."} +{"text": "Nature Communications 10.1038/ncomms14712, published online 8 March 2017.Correction to: The original version of this article contained an error in the author affiliations. Yuan-Xiang Tao was incorrectly associated with Department of Anesthesiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China. This has not been corrected in both the PDF and HTML versions of the article."} +{"text": "Nature Communications 10.1038/s41467-019-10717-9, published online 27 May 2020.Correction to: The original version of this Article omitted from the Genome Aggregation Database consortium the member Marquis P. Vawter, from the Department of Psychiatry & Human Behavior, University of California Irvine, Irvine, CA, USA. Additionally, the following was added to the Author Contributions: \u2018All authors listed under The Genome Aggregation Database Consortium contributed to the generation of the primary data incorporated into the gnomAD resource\u2019. This has been corrected in both the PDF and HTML versions of the Article."} +{"text": "Ludwigia, and the foremost expert on Asian Begonia. He served as associate editor, co-editor in chief, and editor-in-chief of Botanical Studies and its predecessor Botanical Bulletin of Academia Sinica during the period 1992\u20132016. He gathered over 25,000 plant specimens, name 121 plant taxa, and has left a remarkable legacy of literature, collaborations and collections. This article summarizes Dr. Peng\u2019s academic career and commemorates his enduring contribution.Ching-I Peng, the most prolific and internationally recognized Taiwanese plant taxonomist of his generation, passed away on May 1, 2018. Dr. Peng was an eminent worker on the taxonomy of East Asian plants and the genus Ludwigia. During the last decade, he also became one of the most influential Begonia researchers in the world.In the late afternoon of May 1 2018, Dr. Ching-I Peng \u201d is a classic in the field, resulting from tireless and meticulous cytogenetic analyses of thousands of F1 seedlings from artificial hybridizations among species of the section. His work greatly clarified the taxonomy of this notoriously complicated aquatic genus in the southeast USA and the Institute of Botany of Academia Sinica with Dr. Chang-Hung Chou (1978), Ching-I Peng started his PhD study in the summer of 1978 in the joint program of the Department of Biology at Washington University-St. Louis and the Missouri Botanical Garden, working under the supervision of Peter Raven. Ching-I Peng\u2019s dissertation \u201cUSA Peng and reveUSA Peng . Dr. Penata Peng . Throughata Peng and the ata Peng , which fhttps://hast.sinica.edu.tw/), hosting label information, specimen images, and photographs of plants in the field, are accessed by more than 5,000 visitors around the world each month , Brassicaceae and editorial board member for the Flora of China (1997\u20132013). Throughout his career, Dr. Peng organized many domestic and international conferences and edited numerous highly influential conference proceedings, including the first Cross-strait Symposium on Floristic Diversity and Conservation in 1997 and 98 new species of Begonia that resulted in the collection of a deciduous, tuberous and stoloniferous species new to Taiwan. The new species was named B. ravenii C.I Peng & Y.K.Chen to honor Peter Raven , then a PhD student supervised by Cheng-Yih Wu (\u5433\u5f81\u93b0) of the Kunming Institute of Botany, Chinese Academy of Science. In 2002, Shui, Wu, and Peng published \u201cSynopsis of the Chinese species of Begonia (Begoniaceae), with a reappraisal of sectional delimitation project, invited him to contribute to the treatment of Begoniaceae for the FOC.In the summer of 1999, Dr. Peng began to extend his Begonia for the FOC, Dr. Peng\u2019s team travelled to China 15 times, visiting herbaria and type localities, exploring Guangxi, Yunnan, Guangdong, Guizhou, and Hainan, and establishing further collaborations with Chinese botanists. By 2006, 17 new species and one new distribution record of Chinese Begonia were reported into English.To gain first-hand insight into the diversity of Chinese Begonia flora. He soon established a close collaboration with Professor Yan Liu , Indonesia (2 trips), Malaysia (3 trips), the Philippines (5 trips), Thailand (3 trips), and Vietnam (3 trips), and visited public and private collections of Begonia in Australia, Denmark, France, India, Japan, Netherlands, UK, and the United States. BRCAS\u2019s expanding living collection of Begonia became an invaluable asset for both research and conservation, especially with its many collections from type localities . More recently a subset of BRCAS\u2019s Begonia living collection duplicated in the National Museum of Natural Science and curated by Dr. Wei-Hsin Hu \u201d based on his lecture notes, travel logs, and photographs of his calendars to remember Dr. Peng\u2019s ever-lasting passion for Begonia.Dr. Peng also enthusiastically promoted the aesthetic beauty and the conservation of and Peng was publBegonia with more than 100 co-authors, including 35 research articles published in Botanical Studies and its predecessor Botanical Bulletin of Academia Sinica , created and supported by the National Museum of Natural Science (NMNS), The NMNS Foundation, and TSPS, were awarded to encourage and motivate taxonomic studies in Taiwan.Dr. Peng\u2019s official retirement in August of 2015 released him from administrative duties and enabled him to focus fully on the taxonomy of eng Chou , b. In thttps://brmas.openmuseum.tw/), including three exhibitions highlighting Dr. Peng\u2019s work in the flora of Taiwan, Ludwigia, and Asteraceae. An exhibition featuring Dr. Peng\u2019s Begonia research will be released by the end of 2020.To further commemorate Dr. Ching-I Peng\u2019s botanical legacy, the Research Museum of BRCAS is collaborating with the Academia Sinica Center for Digital Cultures to launch a series of on-line exhibitions on the platform of Open Museum (https://www.facebook.com/profile.php?id=1278893812) is still constantly visited and posted by his family and friends around the world.Beyond his considerable academic, research, and professional legacy, Ching-I Peng is remembered by his family, friends, colleagues and students as a kind, generous, positive, and thoughtful person with an engaging sense of humor. He had many interests and hobbies outside of botany, including collecting crafts and antiques of turtles, attending performances of Chinese opera, and playing Ping-Pong at noon-time with his former colleagues of the Institute of Botany. To this day, his personal Facebook page published by Ching-I Peng.Additional file 4. Ching-I Peng\u2019s contribution to the flora of Taiwan: new distribution records (71 species)."} +{"text": "Raquel Laza-Briviesca and Hayley Pearson were not included as authors in the published article. The corrected Author Contributions Statement appears below.SC, AS, and JM conceived and designed the study. SC designed and performed experiments, acquired data, performed statistical analysis, interpreted the data, and wrote the manuscript. DH and RD contributed to the administrative, technical, or material support of the study and critically revised the manuscript for important intellectual content. RL-B and HP carried out extensive assay procedures and results analysis. All authors approved the final version of the manuscript.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Int Braz J Urol, the 3rd under my supervision, presents original contributions with a lot of interesting papers in different fields: Prostate Cancer, Renal Cell Carcinoma, Penile trauma, Bladder Cancer, Neurogenic Bladder, Robotics, Laparoscopy, Male Health, Ureteroscopy, Hypospadia, Urinary diversion and Testicular Cancer. The papers came from many different countries such as Brazil, USA, Turkey, China, Montenegro, Spain, India and Italy, and as usual the editor's comment highlights some of them. Pediatric urology is the highlight of this number. In the present issue we present two important reviews: in page 314 Dr. Kirsch and Arlen from Atlanta - USA (The May-June number of ta - USA present Dr. Azambuja and collegues from Brazil performed on page 353 (Dr. Sharma and Collegues from India performed on page 363 a very iDr. Taha and collegues from Brazil studied patients over 60 years of age, to obtain data on its sexual and urinary health with 3425 questionnaires and found a large number of sexual and urinary disorders and recommended the improvement in health conditions, promoting a better quality of life in the elderly (Dr. Freitas and collegues from Brazil performed on page 383 another Dr. Fedrigon and Collegues from USA performed on page 390 a in vitDr. Barros and Collegues from Rio de Janeiro- Brazil demonstrated in page 409 in a verDr. Kalil and Dr. D'ancona in page 419 (Finally, Dr. Kavaric and Collegues from Montenegro performed on page 446 the studInternational Brazilian Journal of Urology.We hope that readers will enjoy the present number of the"} +{"text": "The typical reliance on self-report questionnaires in retrospective case-control studies of childhood abuse and psychotic disorders has been criticised, due to the potential for recall bias associated with, amongst other factors, cognitive impairments and detachment from reality, among individuals with psychosis. One way to establish if any substantial bias may exist is to examine whether the concordance of reports of childhood abuse established from retrospective self-report methods versus more comprehensive interviewer-rated assessments differ between individuals with psychosis and controls. Data from the Childhood Adversity and Psychosis (CAPsy) study were used to examine the accuracy, strength of agreement, and convergent validity of two distinct retrospective measures of childhood abuse: a self-report questionnaire and a comprehensive interview . In a sample of 234 cases with first-episode psychosis and 293 controls, we found no strong evidence that the validity of the two measures differed between cases and controls. For reports of sexual and emotional abuse, we found fair levels of agreement between CECA and CTQ ratings in both groups (kappa coefficients 0.43\u20130.53), moderate to high sensitivity and specificity, and reasonably high convergent validity (tetrachoric correlations of 0.78\u20130.80). For physical abuse, convergent validity was slightly lower in cases compared with controls. Both measures can be used in future studies to retrospectively assess associations between childhood abuse and psychotic phenomena, but time-permitting, the CECA is preferable as it provides additional important contextual details of abuse exposure. DespiteBias will occur if the validity of recall varies by outcome status . It may be, for example, that recall is less accurate among those with a psychotic disorder because of cognitive impairments , depressIn the absence of a gold standard instrument for measuring childhood abuse retrospectively, some have questioned the convergent validity of different measures . It has It is therefore imperative that we more fully understand, in individuals with psychosis, the degree to which retrospective reports of childhood abuse established from self-report questionnaire-based measures correspond with ratings from interview-administered assessments. Therefore, using data from both patients with first-episode psychosis and population-based controls, we aimed to investigate the: i. accuracy, ii. strength of agreement (as specific types of reliability), and iii. convergent validity of childhood abuse reports obtained using two distinct measures: a self-report questionnaire and an interview-based measure.22.1The sample for this study was drawn from individuals who participated in the Childhood Adversity and Psychosis (CAPsy) study, a population-based case-control study of first-episode psychosis, conducted between 2010 and 2014. Full details of the study, and participant recruitment, are provided elsewhere . Briefly2.2Sections of the Childhood Experience of Care and Abuse (CECA) schedule , an in-d2.3Ethical approval for was obtained from the South London and Maudsley NHS Foundation Trust and the Institute of Psychiatry Research Ethics Committee .2.4Acknowledging the limitations of both self-report and interviewer-based measures of childhood abuse, and that there is no gold standard measure, we examined the level of agreement between the two methods using thAreas under the curve (AUCs), determined from receiver operating characteristic (ROC) analyses, were used to quantify the accuracy of the dichotomised CTQ subscales against the CECA ratings in discriminating individuals with and without moderate to severe levels of abuse. We used the following guideline for evaluating AUC values: <0.70, poor; 0.70 to 0.79, fair; 0.80 to 0.89, good; and 0.90 to 1.00, excellent . Sensiti3Data were available on 234 cases and 293 controls who had completed both the CECA and CTQ. Compared with controls, a greater proportion of the cases were men (64% vs 52%), were less likely to consider themselves as white British (27% vs. 43%), and were younger .The ROC curves for each type of abuse, separately for cases and controls, are shown in Overall levels of agreement (kappa) and sensitivity and specificity are shown in Sensitivity and specificity were fair to high in both cases and controls, with the proportion of individuals who reported abuse on the CTQ and identified as such on the CECA (i.e. sensitivity) highest for emotional abuse. The proportion of individuals who did not report abuse on the CTQ and were identified as such on the CECA (i.e. specificity) was highest for physical abuse in both groups .Finally, we found good convergent validity between CECA and CTQ ratings of abuse in cases and controls for both sexual abuse and emotional abuse . However4In this case-control study, we found that accuracy, strength of agreement, and convergent validity were broadly similar in cases with first-episode psychosis and population-based controls for two distinct retrospective measures of childhood abuse, a brief questionnaire and a more comprehensive interview. Specifically, findings on sexual and emotional abuse showed overall moderate to high proportion of individuals who reported positive or negative histories of abuse on the CTQ compared to the CECA, reasonably high tetrachoric correlations between the measures , and fair levels of agreement of CECA and CTQ ratings in both groups. However, for physical abuse, the proportion of individuals who did not report abuse on the CTQ, but were identified as having been physically abused on the CECA was rather high in cases and controls. Also, convergent validity of self-report and interviewer ratings of physical abuse were marginally stronger in controls. In short, reports of abuse established from these two measures were broadly comparable, and there was no strong evidence that accuracy of abuse established from a self-report questionnaire was lower in cases compared with controls.These findings should be viewed in light of several potential methodological limitations. As for many if not most measures in mental health research, there is no gold standard instrument for childhood abuse, and in the absence of any objective indicators, general bias in retrospective reports due to forgetting, repression of traumatic events, and embarrassment cannot bIn contrast to previous claims that relying on retrospective reports of childhood abuse may affect the validity of findings on the reported associations between childhood adversity and psychosis , our datIn conclusion, the findings reported in this paper provide no strong evidence that the validity of reports of abusive experiences in childhood systematically differs across patients with psychosis and controls. Both of these measures may be used in future research, including case-control studies, of the complex relationship of childhood abuse and psychosis, though time-permitting, the CECA interview may be preferable as it provides more detailed contextual information about the abuse experiences .Charlotte Gayer-Anderson: Investigation, Data curation, Formal analysis, Writing - original draft. Ulrich Reininghaus: Formal analysis, Writing - review & editing. Isabell Paetzold: Writing - original draft. Kathryn Hubbard: Investigation, Data curation, Formal analysis. Stephanie Beards: Investigation, Data curation. Valeria Mondelli: Conceptualization, Writing - review & editing. Marta Di Forti: Conceptualization, Writing - review & editing. Robin M. Murray: Conceptualization, Writing - review & editing. Carmine M. Pariante: Conceptualization, Writing - review & editing. Paola Dazzan: Conceptualization, Writing - review & editing. Thomas J. Craig: Conceptualization, Writing - review & editing. Helen L. Fisher: Conceptualization, Writing - review & editing. Craig Morgan: Conceptualization, Funding acquisition, Writing - review & editing.None."} +{"text": "Aquamavirus A, is currently the only recognized member of the genus Aquamavirus within the family Picornaviridae. The bear picornavirus 1 was recently proposed as the second species in the genus under the name aquamavirus B. Herein, we determined the complete genomes of two novel pinniped picornaviruses, the harbor seal picornavirus (HsPV) and the ribbon seal picornavirus (RsPV). The HsPV and the RsPV were isolated in Vero.DogSLAMtag cells from samples collected from stranded harbor (Phoca vitulina) and ribbon (Histriophoca fasciata) seals. RsPV-infected Vero.DogSLAMtag cells displaying extensive cytopathic effects were processed for transmission electron microscopy and revealed non-enveloped viral particles aggregated into paracrystalline arrays in the cytoplasm. A next-generation sequencing approach was used to recover the complete genomes of the HsPV and the RsPV . Phylogenetic and genetic analyses supported the HsPV and the RsPV as members of the Aquamavirus genus. Based on these results, RsPV represents a novel strain of Aquamavirus A, while the HsPV is a novel strain of the proposed species aquamavirus B. These discoveries provide information on the evolutionary relationships and ultrastructure of aquamaviruses and expands the known host range of those viruses. Our results underscore the importance of the application of classical virology and pathology techniques coupled with high-throughput sequencing technologies for the discovery and characterization of pathogens in wild marine mammals.The seal picornavirus 1, species Picornaviridae is a diverse assemblage of viruses that possess small spherical nucleocapsids and positive-sense RNA genomes ranging between 7 and 8.8 kb in size , Aquamavirus A) and only accepted member of the genus Aquamavirus . The rorovirus; . The SePmavirus , but macavirus; . Furtherellaris) . The cytellaris) .MH760796). The genomes of HsPV and RsPV exhibit low G+C content when compared to other picornaviruses and BePV-1 , which is the causative agent of an important viral hepatitis in humans. The discovery of phopivirus provided insight into the origin and evolutionary history of HAV-like viruses have been detected in the internal organs of pinnipeds and the Asiatic black bear. This suggests that aquamaviruses result in systemic infections , 18. Altin vitro characteristics, virion ultrastructure and morphogenesis, and genetic/phylogenetic analyses supported RsPV and HsPV as novels strains of Aquamavirus A and aquamavirus B, respectively. The results of this study add to a growing body of literature on aquamaviruses and underscores the need for additional research to determine their host range, route(s) of transmission, prevalence, and pathogenicity to terrestrial and aquatic wildlife.In this investigation, we report the complete genome sequences of two novel pinniped aquamaviruses. The The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/supplementary material.The animal study was reviewed and approved by Washington Department Fish Wildlife 109h authority, West Coast Marine Mammal Stranding Network, Alaska Department of Fish and Game (ADF&G) Marine Mammal Permit 15324, NOAA Marine Mammal Stranding Agreement, SR Co-investigator under Permit 932-1905/MA-009526 issued under Section 104 (16 U.S.C. 1375).TR: writing of the original draft manuscript, investigation, formal analysis, and data curation. ON, KB-H, VP, SR, and DL: review and editing of the manuscript, investigation, data curation, and funding acquisition. KS: conceptualization, investigation, formal analysis, software, review, and editing of the manuscript. TW: conceptualization, supervision, funding acquisition, review, and editing of the manuscript. All authors contributed to the article and approved the submitted version.KB-H was owner of the company Alaska Veterinary Pathology Services which is a for-profit diagnostic company. Diagnostic analysis was performed under a contract, however there was no monetary compensation for the production of this manuscript. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "In \u201cDeveloping Theory-Driven, Evidence-Based Serious Games for Health: Framework Based on Research Community Insights\u201d :e11565), an error was found in Reference 2 of the reference list. Arnab S was listed as the author, but should have been listed as the lead editor, instead. The correct author of the book is Kato PM, instead of Arnab S. The correct reference is:Kato PM. The role of the researcher in making serious games for health. In: Arnab S, Debattista K, Dunwell I, editors.Serious Games for Healthcare: Applications and Implications. Hershey, Pennsylvania: IGI Global; 2013:213-231.This correction will appear in the online version of the paper on the JMIR website on April 28, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "The Special Issue (SI) \u201cRemote Sensing in Vessel Detection and Navigation\u201d highlighted a variety of topics related to remote sensing with navigational sensors. The sequence of articles included in this Special Issue is in line with the latest scientific trends. The latest developments in science, including artificial intelligence, were used. The 15 papers (from 23 submitted) were published. Earth observation by multispectral, SAR (Synthetic Aperture Radar), and other sensors provides unique global as well as detailed local surveillance. Resolutions allow for vessel detection, classification, and discrimination from, e.g., icebergs and other objects. Important applications include vessel detection and navigation; trafficking and safety; and monitoring the oceans for fishing, oil spills, territorial violations, piracy, refugee boats, emergencies, AIS (Automatic Identification System) spoofing, etc. With global warming, the north-east and -west passages have opened up for shipping, fishing, and cruise ships in uncharted reef-infested territories littered with sea-ice and titanic icebergs.Vessel detection, classification, and identification;Sea-ice and iceberg detection and tracking;Multi-sensor data fusion;Autonomous ships navigation;Comparative (terrain reference) navigation;Change detection for classifying islands, reefs, and other static objects;Synergy with and comparison to AIS and other vessel identification data;Synergies between satellite sensors with airborne platforms; multiple satellite SAR; optical and thermal infrared sensors including finer resolution sensors, for example, sentinels and other satellites, and in situ measurements;The use of multispectral, multiple frequencies, and polarizations to interpret and quantitatively assess various ocean surfaces, currents, and sea ice phenomena for navigation;Interferometric and Doppler-derived SAR oceanic and sea ice applications focused on surface motion;Validation studies for vessel, ocean, and sea-ice parameters based on in situ and airborne data collections;Use of machine learning and the build-up of annotated training databases;Artificial Intelligence for image data processing.The Special Issue entitled \u201cRemote Sensing in Vessel Detection and Navigation\u201d, was focused on many aspects of multispectral, multi-sensor, SAR, and other sensors related to science/research, algorithm/technical development, analysis tools, synergy with sensors in multiple wavelengths of the e.m. spectrum, synergy with other measurements such as AIS, as well as reviews of the state-of-the-art in-ocean processes using multispectral and SAR imagery for oceans and sea ice, and vessel monitoring for surveillance, trafficking, and navigation. Topics for the Special Issue included the following:In this article we provide a brief overview of the published papers, in particular the use of advanced modern technologies and data fusion techniques. These two areas seem to be the right direction for the future development of vessel detection and navigation.Convolutional Neural Networks (CNN) are state of the art machine learning algorithms for detection and classification of objects in images. This Special Issue includes several works on CNN applied SAR images of vessels.Dai et al. propose Fu et al. investigPan et al. propose Radars are historically the most common method for navigation and vessel detection. There are, however, certain limitations which are addressed by two papers in this Special Issue.Jiang et al. address Lisowski address Video surveillance is becoming increasingly popular in the process of detecting ships at short distances. It is particularly useful in lintel systems both on sea and inland waterways. There are also articles devoted to this issue in the SI.Wawrzyniak et al. present Polap and Wlodarczyk-Sielicka address A number of articles are devoted to the processing of data from various other sensors in the process of detection and eventual identification of floating objects.Ru et al. present Shan et al. address Willburger et al. demonstrDong et al. address The optical spectrum was also investigated in two papers, where special spectrum sensor features could be exploited.2 absorption in the mid-infrared band. Since ship emissions are hot, their CO2 emission spectrum is broader than the subsequent absorption band in the colder atmosphere. As a result, a double peak remains in the IR spectrum that allow for robust remote detection of ships in the mid-IR band.Kim et al. propose Heiselberg demonstrThe next two articles concern unmanned surface vehicles for hydrographic tasks, which have been developing rapidly in recent years pushing out more and more manned ships.Specht et al. address Stateczny et al. address The Special Issue entitled \u201cRemote Sensing in Vessel Detection and Navigation\u201d comprised 15 articles on many topics related to remote sensing with navigational sensors. In this paper, we have presented short introductions of the published articles.It can be said that navigation and vessel detection still remain important and hot topics, and a lot of work will continue to be done worldwide. New techniques and methods for analyzing and extracting information from navigational sensors and data have been proposed and verified. Some of these will provoke further research, and some are already mature and can be considered for industrial implementation and development."} +{"text": "Correction to: Journal of Orthopaedic Surgery and Research (2020) 15:181https://doi.org/10.1186/s13018-020-01653-2Following publication of the original article , it was An error was identified in the Materials and methods section and in Table 3.The updated Materials and methods section is given below and the changes have been highlighted in bold typeface.Materials and methodsrespectively.Twenty-five patients were confirmed as spinal OB in histopathology and treated from January 2008 to December 2018. All procedures were in accordance with the ethics committee of Xijing Hospital and with the Helsinki Declaration of 1975 (revised in 2008). All patients were investigated by the following imaging resources performed in our hospital: plain X-rays, CT scan, MRI, bone scan, and SPECT/CT. Two experienced radiologists and nuclear medicine physicians reviewed the imaging results The correct Table The original article has been corrected."} +{"text": "To compare and rank the clinical effects of different acupuncture and acupuncture-related therapies on patients with hyperlipidemia.We used Network Meta-Analysis (NMA) to evaluate the direct and indirect evidence from relevant studies. Three English and 4 Chinese databases were searched to collect randomized controlled trials (RCT) of acupuncture and related therapies in the treatment of hyperlipidemia. The data were analyzed using Stata15.0 and WinBUGS1.4.3 software after 2 researchers independently screened the literature, extracted the data, and assessed the risk of bias in the included studies.Based on the current evidence, we comprehensively compare the pros and cons of various acupuncture-related therapies, rank the efficacy of various acupuncture-related therapies compared with statins in the treatment of hyperlipidemia, and summarize the best acupuncture intervention methods or combinations.This study will provide new evidence for the safety and effectiveness of acupuncture-related therapies in the treatment of hyperlipidemia, and may be helpful for clinicians, hyperlipidemia patients, and clinical guideline makers to choose the optimal combination of acupuncture for the treatment of hyperlipidemia.INPLASY2020100100 According to the 2016 guidelines for the prevention and treatment of dyslipidemia in adults in China, the prevalence of dyslipidemia among adults in China is as high as 40.40%, a substantial increase from 2002, and the proportion of children and adolescents with hypercholesterolemia in China is also significantly higher. Suggesting that the burden of dyslipidemia and related diseases in China will continue to increase in the future. Hyperlipidemia is closely related to the occurrence of coronary heart disease and stroke, and may induce Alzheimer disease (AD), vascular dementia (VD), and Parkinson disease (PD), etc.,7 At present, statins, beta-blockers, and other lipid-lowering drugs are mainly used in clinical practice, which have some efficacy, but may cause adverse effects such as liver damage, rhabdomyolysis, neoplasia, and diabetes. Due to the complex etiology of hyperlipidemia, it is closely related to environment and genetics, but its mechanism has not been fully elucidated. Acupuncture, as the most representative non-pharmacological therapy in traditional Chinese medicine, without the side effects and clinical contraindications of Western medicine, is a treatment method with potential for development. In recent years, more and more studies have been conducted on the use of acupuncture to treat patients with hyperlipidemia, and previous meta-analyses had found that for patients with hyperlipidemia, acupuncture,\u201311 auricular acupuncture, and herbal medicine may be more advantageous and safer than Western medicine.,10 However, due to the wide variety of acupuncture and the different focus on efficacy, there is still a lack of direct comparative studies between different acupuncture-related therapies. In this study, the Network Meta-Analysis (NMA) method was used to evaluate the effects of various acupuncture-related therapies for patients with Hyperlipidemia, expected to provide evidence-based medicine evidence for selecting the best combination of options.Hyperlipidemia is a disorder of lipid metabolism characterized by an increase in triacylglycerol (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and a decrease in high density lipoprotein cholesterol (HDL-C) in the peripheral blood, and is associated with hypertension, diabetes, and obesity as major vascular risk factors.2 This study has been registered with INPLASY, and registration number was INPLASY2020100100.It will be reported following the Preferred Reporting Items For Systematic Reviews And Meta-Analyses for Network Meta-Analysis Checklist (PRISMA-NMA).2.1Our literature search was performed from database establishment until April 1st, 2020, including 3 English databases: PubMed, EMBASE, Cochrane Library, and 4 Chinese databases: the China Biology Medicine (CBM), the China National Knowledge Infrastructure (CNKI), Wanfang Data, the Chinese Scientific Journal Database (VIP). The search was conducted using a combination of medical subject headings (MeSH) terms and free words. In addition, the references included in the medical literature were retrospectively supplemented to obtain associated references.2.2The published randomized controlled trials (RCT) of acupuncture-related therapies for the treatment of primary hyperlipidemia, regardless of age and gender. Clear diagnostic criteria were required to confirm the diagnosis of primary hyperlipidemia. Interventions in the treatment group were various types of acupuncture-related therapies, including simple acupuncture, electroacupuncture, warm acupuncture, auricular acupuncture, acupuncture point injections, acupoint embedding, or a combination of acupuncture and drugs; the control group is statin lipid-lowering western medicine, or placebo, or comparison between various acupuncture-related therapies. Comparisons investigated are given in Figure 2.31.Research on self-controlled or other non-randomized controlled trials;2.Research with unclear diagnostic criteria;3.Research on subjects belonging to secondary hyperlipidemia ;4.Pre-clinical studies, systematic reviews, case reports, and meta-analysis, etc.;5.The report did not have clear original data, and contacting the author was unsuccessful;6.There was no acupuncture-related therapy or other forms of acupuncture ;7.Repeated research or research report results are the same.2.4Two researchers, ZH and CH, independently conducted the literature screen, data extraction, and cross-check. In any case of disagreement, they would discuss to reach a consensus or a third researcher, CY would assist in the final determination. A unified data extraction table was used for data extraction, which included: including general information of the included literature:1.the name of the first author, journal name, publication year, etc.;2.baseline data of the subjects included in the literature: grouping, the sample size of each group, and Subjects age, etc.;3.intervention methods: type of intervention, treatment frequency and treatment period, etc.;4.risk bias related factors: random method, allocation hiding, blinding, etc.;5.outcome indicators before and after treatment data.2.5,16 However, acupuncture-related therapies were non-pharmacological therapies, therefore some participants and researchers involved in these studies were not able to be blinded. So we required be blinded for the outcome assessment.Our 2 researchers, JL and YW, evaluated the included studies in accordance with the bias risk assessment tool recommended on Cochrane Handbook 5.1.2.6\u201319 TC, TG, LDL-C, HDL-C are numerical variables, and the difference before and after treatment is used as the effect size. In some trials, the change between baseline and after treatment failed to show, and the missing data were estimated using the following formula:Statistical analysis was performed using Stata 15.0 and WinBUGS 1.4.3 software. then WinBugs1.43 was run to set the number of iterations to 50,000 for NMA; 95% confidence interval (95% CI) of inconsistency factors (IF) was used to judge the consistency of the closed-loop. If the IF with 95% CI contains 0, it means that the direct and indirect evidence is consistent, otherwise, it means that there is a higher possibility of inconsistency. Second, the Stata 15.0 program was applied to create funnel plots to determine whether there was evidence of small sample effects in the included studies. Finally, the surface under the cumulative ranking curve (SUCRA) was generated using Stata 15.0 to show the SUCRA scores for all interventions, with higher SUCRA scores implying higher treatment class.First, Stata15.0 was used to draw an NMA evidence relationship diagram;3 Lowering cholesterol is effective in reducing the incidence and mortality of coronary heart disease, and it plays an important role in the prevention and treatment of coronary heart disease. Currently, the drugs used to treat hyperlipidemia have different degrees of adverse effects, and the treatment of lipid-lowering drugs needs to be individualized and tailored, and adverse effects need to be monitored during treatment. It is necessary to pay attention to the adverse effects and regularly test the liver function. And serum creatine kinase, liver function damage, and rhabdomyolysis caused by clinical drug treatment cannot be ignored. Therefore, it is particularly important to find safe and effective treatments for reducing blood lipids.With the improvement of people living standards and changes in lifestyles, hyperlipidemia has become an important public health problem, and its incidence has increased significantly in all age groups. Dyslipidemia is an important vascular risk factor, which is closely related to atherosclerosis and stroke. Studies have shown that in Asian populations, for every 1 mmol/L increase in peripheral blood cholesterol, the incidence of cardiovascular disease will increase by 35%, and the incidence of stroke may increase by 25%.Acupuncture, as a characteristic therapy of traditional Chinese medicine, has a good effect for many diseases in the clinic, which can be used as a safe and effective complementary alternative therapy for hyperlipidemia, considering the safe and side effect-free characteristics of acupuncture operation. It is necessary to scientifically evaluate the curative effect of acupuncture on hyperlipidemia in order to evaluate the curative effect-cost relationship of acupuncture to select the best acupuncture Methods or the best combination of treatments can better reduce the economic burden of patients. This study uses MNA to make up for the lack of direct data. Indirect data is used to compare the efficacy of different acupuncture-related therapies in the treatment of hyperlipidemia, which provides some evidence-based medical evidence for the clinical efficacy of acupuncture-related therapies for hyperlipidemia.Based on the current evidence, we comprehensively compare the pros and cons of various acupuncture-related therapies, rank the efficacy of various acupuncture-related therapies compared with statins in the treatment of hyperlipidemia, and summarize the best acupuncture intervention methods or combinations. Reliable evidence will be obtained for acupuncture-related therapies for the treatment of hyperlipidemia. This study will provide new evidence for the safety and effectiveness of various acupuncture-related therapies as a complementary alternative therapy for hyperlipidemia, it may be helpful for clinicians, hyperlipidemia patients, and clinical guideline makers to choose the optimal combination of acupuncture t for the treatment of hyperlipidemia.Data curation: Xue-Song Wang, Jia-Jia Li.Formal analysis: Chuan He, Zhong-Sheng Huang.Funding acquisition: Li-Hong Kong.Methodology: Xue-Song Wang, Yue-Shen Wang.Project administration: Li-Hong Kong, Miao Wu.Resources: Jia-Jia Li, Yue-Shen Wang.Software: Xue-Song Wang.Supervision: Miao Wu, Li-Hong Kong.Validation: Chao-Chao Yu.Writing \u2013 original draft: Xue-Song Wang, Jia-Jia Li, Yue-Shen Wang.Writing \u2013 review & editing: Chao-Chao Yu; Miao Wu, Li-Hong Kong."} +{"text": "There is currently growing recognition of the complex care needs of patients with life-limiting conditions and their family members, prompting the need to revisit the goals of medicine. This Special Issue reflects a broad research agenda in the field of palliative and end-of-life care. A total of 16 papers of empirical studies and systematic review are included spanning five domains, namely, patient, caregiver, healthcare provider, policy, and methodology. The results generally suggest the merits of palliative care and reveal room for further improvement in palliative care education, manpower, infrastructure, and legal and policy frameworks. The landscape of palliative and end-of-life care has changed substantially in the last decade due to changes in demographics, medical technologies, and disease patterns, resulting in a huge number of people who live with chronic progressive conditions requiring palliative care . Decision = 14) were conducted in Asia (n = 8) and Europe (n = 4), with one in South America and one in North America.This Special Issue intends to inform policy and practice regarding the improvement of palliative and end-of-life care. The articles included reflect multifaceted interests broadly classified into five domains, namely, patient, caregiver, healthcare provider, policy, and methodology, although the studies overlap across some domains . The metFive articles focused on the patient. A wide spectrum of patient populations was included, ranging from cancer patients to noncancer patients and older adults with moderate\u2013severe physical impairment and cognitive frailty. Engel and colleagues reportedTwo studies identified factors of survivorship of palliative care patients using data from routine records. Cheng and colleagues used datThe other two studies in the patient category focused on healthcare utilization. Kao et al. analyzedTwo studies focused on the carer. Leung and colleagues carried Four studies were classified into the category of healthcare workers in palliative and end-of-life care. Lehto et al. intervieThe other two studies in this category related to palliative care undergraduate education. Pieters et al. conducteFour studies focused on policy regarding palliative and end-of-life care. D\u00e1valos-Batallas et al. analyzedOne study was classified in the methodology category. F\u00e0bregues et al. identifiThe studies presented in this Special Issue cover a wide range of issues related to different aspects in the field of palliative and end-of-life care. They document not only the progress being made in different pathways to improve the practices of care and the development and implementation of the related policies in different countries, but also the high complexity of the challenges faced to answer complex questions regarding palliative and end-of-life research. A global shift in the healthcare paradigm from a biomedical-oriented to a holistic approach in multiple and overlapping dimensions is evident, but the progress is still in its infancy stage. Frequent updates from multiple stakeholders could lead to a concerted effort for smoother transition to high-quality palliative and end-of-life care in the future."} +{"text": "This study reviews the research landscape of entrepreneurship studies done by Vietnamese researchers from 2008 to 2018. A sample size of 111 articles from 108 academic outlets indexed in Web-of-Science and Scopus were extracted on the SSHPA database, then read and systematically classified into 15 topics. A systematic review reveals (i) a high frequency of research on various aspects of management, (ii) a lackluster focus on innovation and creativity in entrepreneurial activities, (iii) and worrisome cultural influences on the level of creativity. Overall, there was evidence of a detachment between the academic community and the entrepreneurial community. The research landscape shows there have not been enough studies done on the following aspects of entrepreneurship: technology application, poverty reduction, network development, internationalization, inter-generational transfer, and sex/gender. Entrepreneurship; Entrepreneurship research output; Creativity; Innovation; Economics; Economic development; Business; Management. Bibliom1.2per capita income grew two-fold from USD202 in 1986 to USD417 in 2001, then jumped to over USD2,500 then by 2018 , which is an open database built for the purpose of tracking publications in Web-of-Science/Scopus-indexed academic outlets of Vietnamese researchers in social sciences and humanities. The starting point of data collection is 2008, and it is an on-going national project whose data quality has been validated through a series of publications attempting to find correlates of productivity of Vietnamese researchers. Among the publications, the most notable is the data descriptor paper or the method paper reviewed by Nature Research's Scientific Data, which demonstrates the full potential for reproducibility of the data collecting and cleaning procedure . SDA is 2.2First, the study performs a search within the SSHPA database using the following search keywords: entrepreneurship; entrepreneur; entrepreneurial firms and their synonyms such as small and medium enterprises; small business; startup; micro firms and microfinance. The result is a total of 111 research articles from 108 academic outlets published in the period from 2008 to 2018. All articles\u2019 abstracts were then reviewed by the research team to ensure each article matches the inclusion and exclusion criteria. Admittedly, the search keywords can be widened, which might result in a large sample size. However, given that this study is country-specific, and limited to the period from 2008 and 2018, the number of articles in the sample is comparable to other review studies done in a much larger scale. For example, After double-checking the 111 articles, the authors classified the articles based on proposed a list of eight key topics: Corporate social responsibility; Business efficiency; Innovation and creativity; Gender/sex; Organizational management; Resources Management; and Inter-generational transition. For an article to belong to any one of the above topics, the topic must be analyzed and discussed thoroughly in the articles\u2019 result and discussion section. For example, when a topic is only briefly mentioned then, the article would not be counted toward the topic. Next, the proposed list of topics was refined by two independent researchers who read the full text of the articles and examined any newly proposed topics. Group discussions among research team members are conducted to settle all disputes on inclusion or exclusion of topics. Finally, seven more topics are added: Legal and institutional matters; Internationalization, Entrepreneurial education; Poverty alleviation and Job creation; Network development; Motivations and Values; Technological utilization.10.17605/OSF.IO/NJMSY / URL: https://osf.io/njmsy/) . The attributes of the articles are then coded into the SDA system to generate data visualizations to further check for errors. The dataset is publicly available on OSF (DOI: /njmsy/) . After t33.1Organizational management (discussed in 59 articles), Resources management (53), Legal and institutional matters (39), Business efficiency (38). Clearly, studies over the past decade have built on early scholarship which showed human resources, capital, and government's support were the challenges for Vietnamese entrepreneurs , Corporate social responsibility (38), Social and cultural influences (36), and Motivations and values of entrepreneurs (22). The period from 2012 onwards witnessed a surge of studies on Innovation and creativity, which has remained one of the most investigated topics in this group. There was a noticeably high number of articles concerning social and cultural issues in 2016, followed by a sharp decrease in the following year \u2013 this observation is explained by the overall drop of articles in the field and suggests that the rise was only temporary.The next group examines and outlines the typical features of entrepreneurial behavior in Vietnam, namely Poverty alleviation and Job creation (21), Internationalization (17), Network development (17), Inter-generational transition (14), Education and training (10), Gender/sex (8), Technological application (3). These topics were often only discussed alongside with the main and foundation topics. In general, the number of articles in this group remains low and not consistent. An observable trend here is the slight boost of articles addressing the issue of Network development in 2016, which mirrors the patterns of research about Law, regulation and institutions. This could signify the connections among topics within the field since the issue of Network development often involves discussions about corruption and bribery with implications for institutions and policy.Lastly, the emerging topics concern 3.2As noted in Legal and Institutional matters topic. Legal and institutional challenges for private firms in Vietnam require not only the government to step up but also proactive actions from entrepreneurs. Studies find that policies are inconsistent in nature, prone to changes, and sometimes target at only a small group of SMEs high frequency of research on various aspects of management of enterprises, (ii) lackluster focus on innovation and creativity in entrepreneurial activities, (iii) and worrisome cultural influences on the level of creativity. More importantly, the low level of research output and the existence of several under-researched topics display a lack of engagement between scholar and entrepreneur communities. The wedge between the entrepreneurs and the researchers is reflected most clearly in the low number of entrepreneurship research that tackles technology adaptation and gender-related issues.It is important to notice here the context of development in Vietnam. First, with the development in the infrastructure, both physical and digital, one can expect the situation for entrepreneurship research, especially those related to technological utilization, can quickly take off. Second, universities and research institutions in Vietnam are still trying to catch up with the changes: new technological concepts like DOI, Data storage, preprints, open-access are slowly emerging among scholars in Vietnam . In mediStarting with the research landscape provided in this study, higher education in policy-makers could incentivize research on under-studied topics, namely technology utilization, education and training, and gender/sex issues in entrepreneurship studies. Other areas of studies that should be more encouraged so that entrepreneurship research Vietnam can catch up with the trend of the world are the cognitive and theoretical aspects of entrepreneurship . Built uH.M. Toan: Conceived and designed the experiments; Contributed reagents, materials, analysis tools or data; Wrote the paper.Q.H. Vuong: Conceived and designed the experiments; Analyzed\u00a0and interpreted the data; Contributed reagents, materials, analysis tools or data.V.P. La: Conceived and designed the experiments; Performed the experiments;\u00a0Contributed reagents, materials, analysis tools or data.T.T. Vuong and T.H.K. Nguyen: Performed the experiments;\u00a0Analyzed\u00a0and interpreted the data; Wrote the paper.H.M. Tung: Analyzed and interpreted the data; Contributed reagents, materials, analysis tools or data; Wrote the paper.Vietnam National Foundation for Science and Technology Development (NAFOSTED) under the National Research Grant No. 502.01-2018.19.This research is funded by The authors declare no conflict of interest.10.17605/OSF.IO/NJMSY. URL: https://osf.io/njmsy.The dataset is publicly available in OSF. DOI:"} +{"text": "In the Methods section, a reference is omitted from the second sentence of the first paragraph and the Nottingham Research Ethics Committee project ID is omitted from the third sentence of the first paragraph. The correct sentences are: Full details of the survey methods are published elsewhere . This study received ethical approval from the East Midlands\u2014Nottingham Research Ethics Committee .In the Discussion section, the same reference is omitted from the first sentence of the third paragraph.The correct sentence is: Our previous survey results looking at willingness to reduce alcohol intake for dementia risk reduction showed a positive relationship between having a healthy lifestyle and willingness to reduce alcohol intake .The reference is: Oliveira, D., Jones, K. A., Ogollah, R., Ozupek, S., Hogervorst, E., & Orrell, M. (2019). Willingness to Adhere to Current UK Low-Risk Alcohol Guidelines to Potentially Reduce Dementia Risk: A National Survey of People Aged 50 and Over. DOI: 10.3233/JAD-181224."} +{"text": "Objectives: The worldwide SARS-COV2 pandemic has impacted the health of workers and companies. The aim is to quantify it according to sick leave.Methods: Using ICD-9 codes, we analyzed Ibermutua records of all sick leaves during the first trimester of 2020, compared to during the same months of 2017, 2018, and 2019. We stratified the analysis by causes, patient sex, activity sectors, and regional data. All sick leaves were adjusted by the number of Ibermutua-affiliated persons in each period.Results: In March 2020, there was an unprecedented (116%) increase in total sick leaves, mainly due to infectious and respiratory diseases. Men and women were equally affected. All activity sectors were impacted, with the highest increase (457%) observed among health-related workers, especially due to contagious disease. The incidences of sick leaves were heterogeneous among different regions. Cost-analysis of sick leaves during the first trimester of 2020 compared with in previous years showed 40.3% increment .Conclusions: The SARS-COV2 pandemic is having a huge impact on workers' health, as shown by data regarding sick leaves in March 2020. This is associated with greater economic burden for companies, both due to the cost associated with sick leaves and the losses in productivity due to confinement. Since December 2019, starting in the city of Wuhan, China, the world has been suffering an outbreak of the new coronavirus SARS-COV2, which induces acute respiratory infection, bilateral pneumonia, respiratory failure, and in some cases death and the UK . These cases were followed by a sharp rise in the number of infected individuals. At the end of March 2020, there were 85,195 known cases 1, 2020 a. This raSick leave (SL) is a complex indicator of the working population's well-being and also a predictor for health consequences and mortality , 7. MoreIbermutua is a mutual insurance company in occupational medicine, which collaborates with the National Public Health System in Spain to provide healthcare for the working population. Ibermutua covers over 1.6 million workers, and has nearly 100 of its own health centers and 1,000 external health centers, spread throughout Spain. Regarding non-work-related or common diseases (CD), Ibermutua receives daily information from the National Public Health System about its covered workers who are on sick leave, and the cause of it. This enables assessment of the impacts of CD on health in a large sample of workers, and the associated economic burden on society.The impact of COVID-19 pneumonia on SL and its costs have not been reported in our knowledge, despite of the important component of the economic burden of the disease and the loss of productivity due to morbidity and premature death. In the present study, we aimed to analyze the CD-related SL (CD-SL) of workers during the first trimester of 2020. We compared these data with the findings from previous years, with special focus on health-related workers.We performed a retrospective comparative analysis that included all CD-SL episodes started between January and March 2020 among workers covered by Ibermutua. We compared these data with the CD-SL episodes during the same months in the years 2017\u20132019. Data on these episodes were obtained from the National Public Health System Register for SL due to CD.On January 31, 2020, Ibermutua covered a total of 1,651,305 workers . The mean age was 42 years among all workers, 42.2 years among men, and 41.9 years among women. On this date, the Spanish Social Security covered 19,171,039 workers , which included the workers affiliated with Ibermutua .Ibermutua is a mutual insurance company that provides healthcare for work-related SL episodes and occupational illnesses. They are also responsible for the management of CD-SL for some companies. Mutual insurance companies in Spain collaborate with National Social Security to administer statutory sick pay. Among its preventive activities, it launched the ICARIA (Ibermutuamur CArdiovascular RIsk Assessment) project in 2004 , 12.In the present study, workers were classified into four major activity sectors according to the Spanish Classification of Economic Activities. The sample included workers from all activity sectors and occupational categories, with a broad age range, and from both genders. We performed separate analysis of CD-SL in health-related workers.SL episodes due to infectious disease, cardiovascular disease, respiratory disease, and undefined symptoms were identified based on the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) codes 001\u2013139, 390\u2013459, 460\u2013519, and 780\u2013799, respectively. Additionally, cases of COVID-19 infection were identified using the specific ICD-9-CM codes released on March 6, 2020: non-specified viral infection (codes 079) and exposure to contagious disease (codeV01) . Data are shown as absolute values and percentages. For comparison with the SL in the first 3 months of 2020, to balance the year-to-year variability, we used the mean SL corresponding to the same months of the years 2017\u20132019. Data on common SL were adjusted by the number of Ibermutua-affiliated workers every month, which is shown in The research described herein adhered to the tenets of the Declaration of Helsinki. All medical records were anonymized; only statistical information was provided by Ibermutua for research purposes.At the end of March 2020, there were 136,977 Ibermutua-affiliated workers in the sanitary and social services see . Table 1Finally, the increased SL translated into an 40.3% increase of associated costs during the first trimester of 2020 compared with the same period of 2017\u20132019 . The findings of our present study confirm that in March 2020, there was an unprecedented increase in the number of SL compare with in previous years with the greatest impact on exposed workers . The incIncreasing clinical and epidemiological evidence suggests that SARS-COV2 infection can induce cardiovascular complications that are associated with a worse prognosis , 18. HowConcerning economic activity sectors, most SL (in absolute terms) affected the service sector. However, when data were adjusted by affiliated workers to each sector, the SL increase was lowest in the service sector (85%), and the highest increase was observed in the construction sector (284%). Importantly, this finding indicates that no activity sector was safe. It is interesting to note that many of the SL, especially in March 2020, were among health-related workers. Of the 136,977 health-related workers in our study cohort, 6.78% had a SL, compared to 1.22% in previous years. Additionally, 1,574 SL among health-related workers in March 2020 were coded as due to exposure to biological agents. This code has been very rarely used in past years. These results are in agreement with data on confirmed cases of COVID-19 in Spain, where nurses and doctors comprise almost 20% of affected people up to March 2020 and Catalonia, and higher in the Basque region, Madrid, and Castilla-La Mancha. In some instances, our data showed a mismatch with reports of illness in the general population. For instance, Catalonia had the second highest number of patients affected by COVID-19 pneumonia , but shoOne strength of this analysis of non-work-related (common) diseases, is that Ibermutua receives daily official reports from the National Public Health System of Spain about its covered workers who are on a SL and the cause of it. This encoded information (ICD-9\u2013CM) becomes part of the Ibermutua official records. These data were anonymized and used for the present analyses.As a limitation of this study, it should be noted that during the months of January and February and mid-March, primary care physicians did not have an official regulation for the codification of SL related to the COVID-19 pandemic. This came into use in early March. Thus, especially prior to March, many cases of this infection would have been encoded as undefined symptoms, viral infections, or respiratory infections.In conclusion, in just the first trimester of 2020, the outbreak of SARS-COV2 has had a dramatic impact on the health of the Spanish working population and on their companies. This influence has been especially severe for health-related workers. Moreover, the impact on health is associated with a significant economic burden on society.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.EC-B, MS-C, and PV made the study design and wrote the draft. CC-R, CF-L, and LQ performed all database analyses and performed data collection. AF-M, PM-M, and AG-Q review all the data and make substantial contribution to data interpretation. All authors interpreted the results, contributed to writing the article, and approved the final version for submission.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The complex mechanism of aetiopathogenesis, progress and chronicity of the disease involves genetic, epigenetic and environmental factors. To understand the molecular mechanisms underlying disease phenotypes, one has to place implicated factors in their functional context. However, integration and organization of such data in a systematic manner remains a challenging task. Molecular maps are widely used in biology to provide a useful and intuitive way of depicting a variety of biological processes and disease mechanisms. Recent large-scale collaborative efforts such as the Disease Maps Project demonstrate the utility of such maps as versatile tools to organize and formalize disease-specific knowledge in a comprehensive way, both human and machine-readable. We present a systematic effort to construct a fully annotated, expert validated, state-of-the-art knowledge base for RA in the form of a molecular map. The RA map illustrates molecular and signalling pathways implicated in the disease. Signal transduction is depicted from receptors to the nucleus using the Systems Biology Graphical Notation (SBGN) standard representation. High-quality manual curation, use of only human-specific studies and focus on small-scale experiments aim to limit false positives in the map. The state-of-the-art molecular map for RA, using information from 353 peer-reviewed scientific publications, comprises 506 species, 446 reactions and 8 phenotypes. The species in the map are classified to 303 proteins, 61 complexes, 106 genes, 106 RNA entities, 2 ions and 7 simple molecules. The RA map is available online at ramap.elixir-luxembourg.org as an open-access knowledge base allowing for easy navigation and search of molecular pathways implicated in the disease. Furthermore, the RA map can serve as a template for Rheumatoid arthritis (RA) is a progressive inflammatory and autoimmune disease with unknown aetiology. It affects 0.5\u20131% of the world population, and disease characteristics involve synovial inflammation and hyperplasia, cartilage and bone destruction, production of autoantibodies like rheumatoid factor (RF) and anti-citrullinated protein (ACPA), and various systemic features such as cardiovascular, pulmonary, psychological and skeletal disorders .Systems Biology allows deciphering complex disease mechanisms by treating biological processes in living organisms as coordinated and interdependent events. Especially in human diseases, genes and proteins rarely act alone when affecting implicated cells, tissues or organs. To understand the molecular mechanisms underlying these phenotypes, one has to place the implicated biomolecules in their functional context and interconnect them. This way, a graphical representation of disease mechanisms is established and can be refined, validated and interpreted using the wealth of high-throughput biological data. Nevertheless, integration and organization of both graph and data in a systematic and standardized manner remains a challenge.Molecular maps are widely used in biology to provide a useful and intuitive way of depicting a variety of biological processes and disease mechanisms. Examples of such maps include the gastrin and cholecystokinin receptor signalling , yeast somic data visualization. Omic datasets can be superimposed on the map, pinpointing affected areas in different samples.In this work, we present a systematic effort to construct a fully annotated, expert validated, state-of-the-art knowledge base for RA in the form of a molecular map. The RA map illustrates molecular and signalling pathways implicated in the disease. Signal transduction is depicted from receptors to the nucleus in a systematic fashion using the Systems Biology Graphical Notation (SBGN) standard representation . High-quhttps://bioinfominer.com) and Cytoscape . The researchers of this study used 28 published studies for the construction of the first RA map that included experiments performed in different cell types/tissues/fluids such as the peripheral blood mononuclear cells, synovial fibroblasts, macrophages, chondrocytes, synovial tissues, bone, blood, and synovial fluid (http://www.genome.jp/kegg/pathway.html), and Ingenuity Pathway Analysis (IPA) or were part of well-characterized pathways involved in RA, we used one PubMed or KEGG ID. For the purposes of this project, we aimed to be inclusive of the whole spectrum of RA. In this context, we used RA as a defining criterion and did not make the distinction between sero-negative and sero-positive RA when reviewing the literature.We added annotations for all the components and reactions present in the CellDesigner XML file using the sections text NOTE and Minimal Information Requested In the Annotation of Models (MIRIAM) , which aWe carefully evaluated all elements and reactions of the previous RA map and added annotations concerning experimental validation with small-scale experiments where possible. Molecules, for which we could not find small-scale experiments, were kept if appeared in at least two high-throughput studies. We removed from the map molecules that failed to fulfil the above criteria.http://www.celldesigner.org/plugins.html). The RA map includes six compartments, namely extracellular space, plasma membrane, cytoplasm , nucleus, secreted molecules and cellular phenotypes.To improve the layout of the molecular map, we used the CellDesigner plugin Relayout Model . A separate compartment contains proteins secreted out of the cell and, finally, a dedicated compartment contains cellular phenotypes relevant for RA. The RA map has the form of a cell with surrounding extracellular space, the cytoplasmic area containing organelles, proteins and small molecules, the nucleus with gene-regulatory mechanisms, secreted molecules and cellular phenotypes. We used a distinct colour code for the components in the RA map: plasma membrane receptors in peach, proteins in purple, genes in green, RNAs in red and cellular phenotypes in yellow. Inhibition edges are represented in red colour, while for all others like state transition, catalysis, transport, reduced physical stimulation and heterodimer association we used black colour.Experts\u2019 curation is critical to reconstructing molecular and cellular interactions from the available literature. Due to the complexity of RA regarding cell types , mediators of inflammation and the variety of biological processes implicated in the disease, the review of the map by RA experts was necessary for an accurate representation of disease hallmarks. To provide a systematic and comprehensive molecular map, we used SBGN standards and a cell layout. We took advice from experienced scientists in both biological and computational domains to make the content comprehensive and functional for different types of users such as experimental biologists, clinicians, computational modellers and bioinformaticians. The RA map layout, the representation of various levels of information and the validity of molecules and pathways included in the RA map, were carefully examined in this context.https://royludo.github.io/cd2sbgnml) for converting the CellDesigner XML file into SBGN-ML format and subsequently import the file to VANTED for further analysis.The SBGN is a staomic datasets (see https://www.drugbank.ca/), CHEMBL (https://www.ebi.ac.uk/chembl/), CTD (http://ctdbase.org) and miRTarBase (http://mirtarbase.mbc.nctu.edu.tw).The RA map is available as an online interactive map using MINERVA (Molecular Interaction NEtwoRks VisuAlization) platform . MINERVAsets see . Moreovesets see , CTD to the plasma membrane (ligand\u2013receptor complexes) and then to the cytoplasm , the nucleus (gene regulation) and the secreted compartment or cellular phenotypes .(i) Cytokines and chemokines: a diverse group of proteins like tumour necrosis factor (TNF) and interleukins to list a few, implicated in various phases of RA pathogenesis by promoting autoimmunity, initiating and maintaining chronic inflammatory synovitis and driving cartilage and bone destruction ;(ii) Growth factors: such as epidermal growth factor (EGF), fibroblast growth factor (FGF), insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), activate intracellular signalling pathways (such as PI3K-AKT pathway) and regulate a broad range of cellular functions like cell growth, survival, cell motility and apoptosis , 51;(iii) Toll-like receptors (TLRs): TLR2 and TLR4 are primarily expressed in synovial fibroblasts and macrophages in human RA joints . ActivatThe RA map contains hallmark cellular and molecular pathways that participate in disease pathogenesis. In signalling cascades, the activation occurs as a response to an upstream stimulus. After activation, the signal propagates through a series of coupled reactions from the plasma membrane to the cytoplasm, to regulate key factors that are responsible for gene regulation and different cellular phenotypes. The RA map includes the following upstream stimuli:(i) The JAK-STAT pathway: this is an effective target in RA therapy. Many cytokines, including IL-6 and TNF, which are validated therapeutic targets in RA, activate directly (for example IL-6) or indirectly (for example TNF) this pathway by phosphorylating JAK proteins. JAKs, in turn, phosphorylate STATs, which then dimerize and translocate to the nucleus and bind to regulatory elements of DNA modulating the expression of target genes , 58. Act(ii) The NF-\u03baB pathway: it is involved in inflammation, cell survival and proliferation. Activated NF-\u03baB is detected in immune cells (such as macrophages and lymphocytes) as well as in stromal cells and stimulates the transcription of arthritogenic mediators like IL-1, TNF, RANKL, PTGS2 and IL-6 in RA synovium. TNF, IL-1 and RANKL are key upstream RA-relevant triggers of the activation of the NF-\u03baBpathway .(iii) The MAPK pathway: all the three classes of MAPKs, namely ERK, JNK and p38, are found to be expressed and activated in synovial tissue in RA. A series of cytokines including among others TNF, IL-1 and IL-6 activate ERK, JNK and p38 MAPK in synovial tissue with successive induction of proinflammatory mediators such as cytokines and tissue destructive enzymes (e.g. MMP-1 and MMP-13) , 62 Nega(iv) The PI3K-AKT pathway: growth factors like VEGF and FGF induce the PI3K-AKT pathway , 63\u201365. The activation of these upstream components leads to the activation of downstream pathways that include:All signalling cascades end at specific cellular outcomes grouped in eight distinct phenotypes such as inflammation , 66, 67,ramap.elixir-luxembourg.org in the form of an interactive diagram, using the platform MINERVA (Molecular Interaction NEtwoRks VisuAlization) (https://www.drugbank.ca/), CHEMBL (https://www.ebi.ac.uk/chembl/), CTD (http://ctdbase.org) and miRTarBase (http://mirtarbase.mbc.nctu.edu.tw).The RA map is available at ization) . ClickinRA map offers custom visualization and export capabilities via MINERVA plugins . For insThe RA map contains information from various sources serving as a generic blueprint for disease mechanisms. However, due to extensive annotation and reference, the user can opt for visualizing cell-specific nodes and interactions. In the RA map, we have grouped our sources in seven distinct groups: synovial fibroblasts, synovial tissue, peripheral blood mononuclear cells (including PMNs), blood (including T and B cells), synovial fluid, chondrocytes and macrophages . SynoviaWe used publicly available datasets for visualization with the RA map. Our goal was to compare the differentially expressed pathways or map regions in different datasets. For this purpose, we used the datasets from transcriptomic data of synovial tissue . We perfhttps://bioinfominer.com) to perform a functional analysis of the RA map. The application performs a biological interpretation of gene sets, which comprises detection and prioritization of systemic processes and pathways, as well as prioritization of genes based on their mapping to those processes. We used BioInfoMiner as a second layer of analysis to see if the functional enrichment would give results relevant to the autoimmune process and RA. We performed two sets of analyses using gene ontology (GO) and human phenotype ontology (PHO) terms. The first analysis using GO gave enrichment of terms like Inflammatory response, Regulation of cytokine production and Activation of MAPK activity, all relevant to pathways included in the RA map. The top five GO terms included apoptotic signalling pathway, positive regulation of cell death, negative regulation of apoptotic signalling pathway, positive regulation of NF-kappaB transcription factor activity and regulation of I-kappaB kinase/NF-kappaB signalling. It also gave a list of 48 prioritized genes . In directed networks such as signalling networks where the reactions are oriented (i.e. from the ECM to the nucleus) we can distinguish two types of node degree: the in-degree, meaning the number of directed edges that have the node as target, and the out-degree that is the number of directed edges that have the node as source. Node degree is an individual characteristic for each node, but a degree distribution can be computed to assess the diversity of the whole network.The majority of biological networks display scale-free properties , which mFirst, we performed the analysis considering the network as undirected to obtain the overall degree distribution (in and out) and then as directed to get the in-degree and out-degree distributions. All node degree distributions follow a power law, showing that the RA network is indeed a scale-free network Figure .In Visual representation of complex pathways and biological processes involved in a disease allows clinical and life sciences researchers to explore relevant mechanisms, which are often intricate and intertwined. Standardized representation and formalization of knowledge in the form of disease maps create an interface to a broad range of bioinformatics and modelling workflows. We present here a state-of-the-art, large-scale molecular interaction map for RA, which is to our knowledge the first SBGN-compliant Process Description disease map. While other efforts, such as the Asthma map, follow the SBGN format, their approach is different as they use three levels of granularity and different SBGN representations for every layer of information. The Process Description level for Asthma map consists of a set of separate modules that correspond to an Activity Flow layer, while the RA map is a global Process Description disease map.http://disease-maps.org/). The community fosters the exchange of good practices and promotes the use of standards for the development of disease maps. The standards of curation and graphical representation, as well as the extensive annotation in both human and machine-readable formats of the RA map, ensure transparency, reproducibility and reusability of its content.All the components and reactions are annotated using only RA and human-specific studies. The RA map is part of the Disease Maps Project, a large scale community effort to comprehensively represent mechanisms for various diseases , 14 described in 28 studies combined with data available in the KEGG database. A total of 435 species , 265 reactions and 10 phenotypes involved in RA were identified using this approach. We decided to follow a different approach as described in the methodology section, in an attempt to limit false positives, increase confidence by incorporating experts\u2019 advice and promote the use of SBGN standards for representation to assure reusability of the map. The new RA map we present here includes information from 353 peer-reviewed publications, and it has a significantly bigger size, as it features 506 species, 446 reactions and 8 phenotypes. The species in the map are classified to 303 proteins, 61 complexes, 106 genes, 106 RNA entities, 2 ions and 7 simple molecules.The RA map can also be used as an interactive knowledge base, using the platform MINERVA and serve as a template for overlaying multiple datasets. Visualization of experimental data could help highlight aspects of the affected biological process and make differences between experimental conditions more evident. Visualizing the results of differential expression analysis of three datasets of gene expression of RA synovial tissues showed enrichment in all cellular phenotypes but not in apoptosis. This finding is in line with the fact that fibroblasts, which constitute a large percentage of the RA synoviocytes, have an apoptosis-resistant phenotype , 108.We performed functional analysis and gene prioritization using BioInfoMiner . The genThe RA map serves as a curated knowledge base, but it can also be analysed as a complex network. Topological analysis can reveal underlying structural features of the RA map like unconnected parts of the network, or important hubs (well-connected nodes) which are otherwise hard to perceive in large-scale networks. The topological analysis performed in this study revealed connected and unconnected parts of the network. This result reflects our fragmented knowledge on the one hand, but also the use of stringent criteria for the nodes included in the map: experimentally validated interactions in at least two published studies, use of data of strictly human origin and disease-specific.Another reason that contributes to the limited wiring of some of the RA map components is the unavailability of known interactions for newly discovered factors for RA. However, we keep them present because the RA map also works as an encyclopaedia for the disease, even if some parts of the puzzle are still missing.et\u00a0al., based on high-throughput data.The topological analysis also assists in the understanding of significantly connected nodes (hubs), placing them in their functional context. The top ten hubs of the RA map as seen in omic datasets and finally, to computational modellers a mechanistic scaffold for the inference of a computational model . Right clicking on the main screen also gives an option to export the visible content in three formats \u2013 SBML, CellDesigner SBML and SBGN-ML.The authors declare that they have no competing interests. G.D.K. is currently also an employee at Regeneron Pharmaceuticals Inc. and declares no conflict of interest regarding the content of this manuscript.AN designed the study, A.N., V.S. and G.D.K. built the new RA map content, V.S. drew the map including the addition of annotations and performed literature mining and topological analysis. G.D.K. validated the content, V.S. and M.V. performed DEG in datasets, M.V. produced scripts for all graphs, A.N. and E.P. performed functional and systemic analysis with BioInfoMiner, V.S. and A.M. validated the SBGN Process Description format of the RA map, M.O. and P.G. set up the MINERVA online version of the RA map, M.O. and V.S. added annotations to reactions of the RA map, V.S. and E.B. produced IPA RA datasets, E.P.T. helped with the validation of functional analyses results and A.N. and V.S. wrote the manuscript, all authors read and suggested modifications, all authors read and approved the final manuscript. Corresponding author: Dr Anna Niarakis.ra-figS1_baaa017Click here for additional data file.ra-figS2_baaa017Click here for additional data file.ra-figS3_baaa017Click here for additional data file."} +{"text": "The number of aging individuals is growing and, along with it, a subset of the oldest-old (those over 85 years), including centenarians. Although researchers have begun identifying issues and needs related to this population , still little is known about decision-making processes as they relate to housing. In rural areas, in specific, centenarians are limited by few residential choices and lack of geographic mobility. In this study, decision-making processes are examined, with an emphasis on interactions between aging individuals and their rural family caregivers. In addition, since family caregivers typically experience a pattern of burnout over time , a second focus of the study is caregiver stress. Data for the study are drawn from semi-structured interviews with a sample of family caregivers in the Midwest. All caregivers had a 100-plus family member recently placed, or in process of placement, at a residential long-term care facility. To meet criteria, all facilities were in towns of 4000 individuals or less. Data consisted of qualitative interviews with the primary family contact , and were analyzed according to Strauss & Corbin (1990). Decision-making themes centered on health and family pressure. Stress themes centered primarily around work. Data are discussed in terms of family strengths, health and wellness, and the need for continued programming for family caregivers, particularly in rural areas."} +{"text": "Background: Gambling disorder (GD) is the most common behavioral addiction and shares pathophysiological and clinical features with substance use disorders (SUDs). Effective therapeutic interventions for GD are lacking. Non-invasive brain stimulation (NIBS) may represent a promising treatment option for GD.Objective: This systematic review aimed to provide a comprehensive and structured overview of studies applying NIBS techniques to GD and problem gambling.Methods: A literature search using Pubmed, Web of Science, and Science Direct was conducted from databases inception to December 19, 2019, for studies assessing the effects of repetitive transcranial magnetic stimulation (rTMS) and transcranial direct current stimulation (t-DCS) on subjects with GD or problem gambling. Studies using NIBS techniques on healthy subjects and those without therapeutic goals but only aiming to assess basic neurophysiology measures were excluded.Results: A total of 269 articles were title and abstract screened, 13 full texts were assessed, and 11 were included, of which six were controlled and five were uncontrolled. Most studies showed a reduction of gambling behavior, craving for gambling, and gambling-related symptoms. NIBS effects on psychiatric symptoms were less consistent. A decrease of the behavioral activation related to gambling was also reported. Some studies reported modulation of behavioral measures . Studies were not consistent in terms of NIBS protocol, site of stimulation, clinical and surrogate outcome measures, and duration of treatment and follow-up. Sample size was small in most studies.Conclusions: The clinical and methodological heterogeneity of the included studies prevented us from drawing any firm conclusion on the efficacy of NIBS interventions for GD. Further methodologically sound, robust, and well-powered studies are needed. Gambling disorder (GD), also known as pathological gambling, affects people of all ages and is a major clinical issue associated with reduced quality of life, psychiatric comorbidity, cognitive deficits, and higher risk of suicide was previously classified as an ICD but is currently considered the prototypical example of behavioral addiction and is included in the diagnostic category of substance-related and addictive disorders according to the The impairment of dopaminergic brain reward circuitries, which are supposed to play a key role in SUD has been explored for the treatment of GD and other behavioral addictions recommendations or para-clinical outcomes .The Pubmed, Science Direct, and Web of Science databases were searched for peer-reviewed studies on NIBS techniques in subjects with/or at risk of GD or pathological/problem gambling and published from databases inception until December 19, 2019. Only studies written in English were considered.The search string for Pubmed and Web of Science was: AND .The search strategy for Science Direct database included: (Gambling OR gamblers) AND (NIBS OR non-invasive brain stimulation OR brain stimulation), then (Gambling OR gamblers) AND , and (Gambling OR gamblers) AND .Two authors (CZ and EM) independently screened titles and abstracts using Rayyan software independently extracted the following data: study design , sample size, gender, presence of any comorbidity with GD , type of rTMS/t-DCS protocol , targeted brain area, outcomes of interest .A descriptive analysis of the results was carried out, focusing on the effects of the interventions. A meta-analysis was not possible due to the small number of studies and subjects, and to the clinical, methodological , and outcome heterogeneity of the included studies.A total of 400 records were identified. After removal of duplicates, 269 papers were screened through title and abstract and 13 papers were obtained for full-text screening. The reference lists of the relevant papers were inspected for additional studies potentially missed in the databases search, but no significant papers were further added. Two authors (CZ and EM) independently evaluated the 13 papers selected for the full-text examination. Disagreement was solved by consensus between the two reviewers, therefore the third reviewer's (ST) advice was not required.Eleven studies met the inclusion criteria and were therefore included in the systematic review .The included papers evaluated the efficacy of NIBS interventions based on rTMS or t-DCS techniques for subjects with GD or problem gambling. Studies were grouped according to the NIBS technique employed and the presence or absence of a sham-NIBS control arm.Seven studies employed rTMS in GD .Zack et al. assessedGay et al. performeIn the study conducted by Sauvaget et al. , one sesIn an open-label study that explored the effect of 15 sessions of low frequency rTMS over the left DLPFC in five participants with GD, despite initial improvement in rating scales, the effect decayed over time and the authors concluded that rTMS treatment failed to demonstrate effectiveness , reported no significant changes .Another question that should be explored is whether NIBS is effective as stand-alone or add-on treatment . Finally, methodologically sound and well-powered double- or triple-blind randomized controlled studies, including clinical outcomes and surrogate biomarkers, are needed to document the potential therapeutic role of NIBS in GD.The datasets generated for this study are available on request to the corresponding author.This study has been designed by CZ, EM, AF, FL, and ST. Data have been gathered by CZ and EM, under the supervision of ST. Data have been analyzed by CZ and EM. The manuscript has been drafted by CZ, EM, AF, and ST. FL and ST revised the manuscript. All authors approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The study aimed to identify the association between the lifestyle-related factors and the cancer-specific, or non-cancer-specific mortality, when accompanied by a competing risk. Two statistical methods were applied, i.e., cause-specific hazard (CSH), and sub-distribution hazard ratio (SHR). Their respective key advantages, relative to the actual study design, were addressed, as was overall application potential.Source data from 4,584 residents (34.2% men), aged 45\u201364 years, were processed using two different families of regression models, i.e., CSH and SHR; principal focus upon the impact of lifestyle-related factors on the competing risk of cancer and non-cancer mortality. The results were presented as hazard ratios (HR) with 95% confidence intervals (95% CI).Age, smoking status, and family history of cancer were found the leading risk factors for cancer death; the risk of non-cancer death higher in the elderly, and smoking individuals. Non-cancer mortality was strongly associated with obesity and hypertension. Moderate to vigorous physical activity decreased the risk of death caused by cancer and non-cancer causes.Specific, lifestyle-related factors, instrumental in increasing overall, and cancer-specific mortality, are modifiable through health-promoting, individually pursued physical activities. Regular monitoring of such health-awareness boosting pursuits seems viable in terms of public health policy making. According to World Health Organization (WHO), up to 30%\u201350% of cancer-related deaths may be precluded through application of effective preventive strategies and elimination of risk factors . In geneCompeting risk analysis, which complements traditional survival analysis, helps evaluate the risk of a specific endpoint when accompanied by the competing events . The most survived time e time t . The effe time t . One of e time t . Therefoe time t .The study aimed therefore to identify the association between the lifestyle-related factors and the cancer-specific, or non-cancer-specific mortality, in the presence of a competing risk.The PONS Project, i.e., \u201cEstablishment of infrastructure for population health research in Poland\u201d aimed to collect population data, with a view to assessing the determinants of health and the main causes of morbidity and mortality in Poland. Sixty thousand local residents, aged 45-64 years, were selected from a single urban district \u2013 of whom 13% were included in the PONS sample, and 50,000 residents from a single rural district \u2013 of whom 10% were included in the PONS sample. During the recruitment period, 12% of the target population were recruited to the PONS study, including 4,799 urban residents. The study protocol comprised a Health Status Questionnaire, medical examination, basic anthropometric measurements, and blood and urine sampling. The Health Status Questionnaire addressed the psycho-social determinants of health status on interactive, structural, and behavioral levels .Pursuant to applicable legislation regulating access to the PONS data, only the data pertaining to permanent urban residents were used in the present study. The verification covered the data of 4.799 (33.7% of men) survey participants. In order to evaluate association of select confounders with cancer death, all participants diagnosed with cancer (n=215) at baseline were excluded. In the last step, 4,584 of participants were covered by the final analysis.The follow-up spanned the period since the date of clinical examination (September 2010\u2013December 2011) until the date of death (caused by cancer or non-cancer causes), or the data censoring point, set for April 2018. The mean follow-up time in the group of 4,584 participants reached 7.6 years.Cancer-specific mortality was the primary outcome, whereas secondary endpoint comprised non-cancer-related deaths. The cause of death was established against the local Cancer Registry (CR) and death registry. The International Classification of Diseases and Related Health Problems (ICD-10) was applied to report the primary cause of death. Cancer causes of death were defined by the following codes: C15, C16, C18, C20, C22, C24, C25, C34, C43, C50, C52, C54, C56, C64, C67, C71, C83, C84, C90-C92. The most frequently observed cause of death from cancer (34.0%) was the lung cancer.Four measures of general obesity and fat distribution were applied, i.e., body mass index (BMI), waist circumference, waist-to-hip ratio, waist-to-height ratio. Body weight was measured with an accuracy of 0.1\u00a0kg using body composition analyser Tanita SC-240 MA, while height was assessed in an upright position by a Seca height measure (with an accuracy of 0.1\u00a0cm). BMI was calculated as the ratio of the body mass (in kg) divided by the squared height (in metres). Natural waist indentation or umbilicus were used as specific points against which the waist circumference was measured. The hip circumference was measured at the widest part of the hips. Waist-to-hip and waist-to-height ratios were calculated as the ratio of waist circumference divided by the hip circumference and the waist circumference divided by height, respectively. Self-reported alcohol and red meat intake were specified in grams per week. Systolic and diastolic blood pressure were measured by a blood pressure monitor Omron (Model M3 Intellisense) and calculated as the average of two readings executed by medical personnel.Serum measurements were taken using enzymatic methods in a laboratory, in compliance with pertinent reference standards. The concentrations of fasting glucose level, total cholesterol, high density lipoprotein (HDL-C) and triglyceride (TG) were obtained by means of the enzyme method with hexokinase, cholesterol oxidase, and cholesterol esterase methods, as well as by making use of the direct method with TOOS and surfactant, respectively. Laboratory tests were carried out with the aid of CB 350i Wiener Lab.All the variables listed so far, as well as age and comorbidities, were treated as the continuous variables.Smoking status was deemed a strong and widely acknowledged confounder of cancer death. Level of education , marital status (single and in a relationship) and occupational activity (inactive and active) were used as a measure of socioeconomic status, and treated as factor variables. Self-reported moderate to vigorous physical activity in leisure (MVPA) was calculated as a continuous variable in minutes per week, based on the replies furnished by the respondents in the long version of the International Physical Activity Questionnaire. Self-reported comorbidities and familial cancer history were treated as the factor variables. Basic characteristics of the study population are presented in The percentage of missing values of each variable were reported. Participants with missing values \u22641% were excluded from further analysis. None of the analyzed variables had missing values >1%. Two different families of regression models in the presence of competing risk factors were applied. The Cox model described the effect of covariates on the cause-specific hazard of the outcome, and the Fine and Gray model addressed the subdistribution hazard function. Both models accounted for the presence of competing risk, but required different interpretation. The regression coefficient of a cause-specific hazard model (CSH) presented the relative effect of a specific covariate on the relative change in the rate of occurrence of the event of interest in the subjects who were currently event-free. The subdistribution hazard ratio model (SHR) made it possible to estimate the effect of covariates on the cumulative incidence function for the event of interest. With a view to ensuring numerical stability, all continuous variables were centred and scaled. All regression models, both unadjusted and adjusted for any identified confounders, i.e., age, smoking status categorized as current and non-current smoker (never and former smoker), and a history of diabetes, were stratified by sex. The results of each model estimation are presented as hazard ratios (HR) with 95% confidence interval (95% CI). All statistical analyses were completed with the aid of R version 3.5.3 software package.A sensitivity analysis was carried out for the associations of analyzed covariates with cancer death and non-cancer death. The three different Cox models with the same set of predictors were estimated for each covariate. For the study model, independence of events was assumed. Cancer death was treated as an event, whereas an occurrence of a competing event (non-cancer death) was treated as the censored observation. In the extremal model 1, the event of interest (cancer death) and a competing event (non-cancer death) were treated equally, and modelled simultaneously. In the extremal model 2 cancer death was treated as an event of interest, while the individuals in whom the competing event occurred, were attributed the longest period of observation within the group, and treated as the censored observations .Age and smoking status were also associated with non-cancer death. An increase in the age equivalent to 1 SD increased the risk of non-cancer death by 1.3 times . Current smokers had a two-fold higher risk of death from non-cancer causes than never and former smokers . History of diabetes increased the risk of non-cancer death more than twice. Strong associations of adiposity markers with non-cancer mortality were noted. Out of all above-referenced indicators, those appeared strong predictors of death. In both sexes, an increase of systolic and diastolic blood pressure equal to 1 SD proved to increase the risk of death by 1.5 and 1.4 times, respectively. Upper level of education and living in a relationship reduced the risk of non-cancer death. Similarly, an increase of MVPA by 1 SD (52.8 min/week in men and 41.6 min/week in women) was associated with a 60% risk reduction of non-cancer death. The values of hazard ratios for cancer and non-cancer death were comparable after adjustment for age, smoking status, and a history of diabetes. Occupational activity was shown to reduce the risk of cancer and non-cancer death by 60%. The above-referenced association is likely to result from the fact that occupational activity is more common amongst the younger persons, thus indicating the age to be the main risk factor for both cancer and non-cancer death.Based on the sensitivity analysis Figure 1In order to assess the real impact of a specific risk factor, e.g., tobacco smoking, on the incidence of an event under study, i.e., lung cancer death, a time-machine would come very handy, let alone the need to have a number of ethical rules broken. Under such perfect conditions, a select group of individuals should then be forced to smoke tobacco for 10 years, so that in due course an incidence of an event of interest might be assessed. Then, we would need to have the time-machine go back in time with the same group of individuals aboard, have smoking tobacco strictly banned, and then have the event of interest re-assed after 10 years. Fortunately enough, applicable laws of physics are not subject to any voluntary suspension, nor are any bioethics committees inclined to go along with such an unorthodox study design, even though these are not the only restrictions to be dealt with.Assessing the actual impact of a specific risk factor on the incidence of a health event is often challenging, as the same risk factors are appreciably instrumental in the incidence of discrepant health events which also happen to compete with each other. Tobacco smoking is both a risk factor for fatal and non-fatal CVDs, as well as for fatal and non-fatal cancers . ConsequThis said, if the main aim of the analysis consists in assessing the actual impact of the variable under study on the duration, then making use of the Cox proportional hazard model is the right thing to do. On the other hand, by taking advantage of the fact that the Cox models maintain the assumption of independence of censoring and of the event under investigation, it is feasible to estimate the actual impact of the predictors on each one of the events under investigation separately, against their modelling. A modification of the Cox model for competing events is offered by the Lunn-McNeil model, which allows for modelling the events under study within a single model . This moMaking use of the PONS study cohort, the association of select, lifestyle-related factors with cancer-specific and non-cancer-specific mortality was assessed. Considering that the same lifestyle-related factors, e.g., obesity, smoking, alcohol, dietary intake of red meat, and low level physical activity, are the acknowledged risk factors for both cancer and non-cancer death , making use of the statistical methods which take due account of the competing risks has deliberately been opted for , 9. To tAge, smoking status, and a family history of cancer were believed the leading risk factors for cancer-specific mortality. Likewise, the risk of non-cancer death was higher in the elderly and smokers. Overall obesity and the distribution of adipose tissue, as well as elevated systolic and diastolic blood pressure, were strongly associated with non-cancer mortality. MVPA in leisure time, and occupational activity were believed to be appreciably instrumental in reducing overall risk of all-cause mortality. The unadjusted hazard ratios (0.37), and adjusted (0.70) for the association of occupational activity and cancer death indicated there was a confounder factor. The age co-variable present in the adjusted model significantly increased the risk of cancer death, while at the same time modifying the association between occupational activity and cancer death. Significance of the age variable as a confounding factor was not encountered in the analyses of the association between occupational activity and non-cancer death, though. Obesity is strongly associated with fatal and non-fatal cardiovascular events . NeverthThe impact of age on overall, and cancer-specific mortality seems to be fairly obvious. The highest incidence of cancer is noted in the sixth decade of life. According to Eguchi et\u00a0al. 2/3 of lMost of the lifestyle-related factors, as assessed in our study, may be modified through individual motivation to have a more health-promoting attitude adopted in daily living . It is wOn the other hand, we strongly believe that geographical, cultural, and political differences shaping health-promoting attitudes within a society are well-worth highlighting. Political transformation of Central and Eastern European Countries over the past 30 years has brought about profound, mostly adverse social changes, including the emergence of a consumer society. The issue of obesity, steadily on the rise, notably absent in the countries under the communist rule, has spawned an increased incidence of the so called lifestyle diseases, mostly cardiovascular disorders and cancer , 43. ObvOne of the study limitations consists in a relatively short follow-up period, corresponding to a relatively low number of cancer- and non-cancer-specific deaths. In pursuance of the Act on Personal Data Protection (GDPR), the authors did not have access to the medical records on the non-cancer causes of death. This is a true limitation of the study, although diligently taken into account, when interpreting and discussing its outcomes. Even though overall nature of the lifestyle-related factors might well seem indicative of a strong likelihood of a cardiovascular cause of death, this cannot be confirmed beyond reasonable doubt, as specific cause of death may not be freely accessed. The data regarding select risk factors were based upon the respondents\u2019 questionnaires only, hence appreciable potential for certain inaccuracies in the conclusions drawn on the associations under study. The PONS cohort was selected through the deliberate sampling, within the 45\u201364 years age range constraints. The fact that no random sampling was used may indeed be deemed a certain limitation in generalising the outcomes of the present investigation, or as burdened with specific age limit constraints with regard to the population under study.Age and smoking strongly affect the risk of either cancer- or non-cancer-specific mortality. Overall content and distribution of adipose tissue, as well as arterial hypertension, were associated with non-cancer-specific mortality. Regular physical activity of moderate or vigorous intensity decreased the risk of death caused by cancer and non-cancer causes. All of the lifestyle-related risk factors, deemed instrumental in increasing overall and cause-specific mortality, are modifiable. Systemic implementation of specific methods aimed at their effective control appears essential in terms of public health interest, and should therefore be prioritized, when mapping out preventive initiatives targeted at the high-risk groups.pawel.macek@onkol.kielce.pl.The datasets presented in this article are not readily available, as the proprietary rights to them are held by the Holycross Cancer Centre (HCC). The Authors, as staff members of HCC, may nevertheless freely access them at any time for research purposes, on a free-of-charge basis. The Authors may also make a certain part of those data sets available to academic researchers, following their prior conversion into an anonymized format, when approached with a reasonable request, care of the First Author. Requests to access the datasets should therefore be directed to Dr Pawel Macek at The PONS study was approved by the ethics committee within the Cancer Center and by the Institute of Oncology in Warsaw, Poland. The present study was duly approved by a local Ethics Review Committee, Faculty of Health Sciences , The Jan Kochanowski University (JKU) in Kielce, Poland.Conceptualization, PM, MB, MT-D, JS-K, HK, EN, SG, and MZ. Methodology, PM, EN, and MZ. Software, PM and JS-K. Validation, HK and MZ. Formal analysis, PM, MT-D, and EN. Resources, MM, MB, and JS-K. Data curation, PM, MT-D, and MB. Writing\u2014original draft, PM, MB, MT-D, and MZ. Writing\u2014review and editing, MM and MZ. Visualization, PM, MS, and JS-K. Supervision, MM, SG, and MZ. Project administration, PM, SG, HK, and MZ. Funding acquisition, HK and MZ. All authors contributed to the article and approved the submitted version.The Project is supported under the programme established by the Minister of Science and Higher Education - \u201cRegional Initiative of Excellence\u201d - spanning the period 2019\u20132022; Project No 024/RID/2018/19; amount of financing allocated: PLN 11999 000.00.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Joanna M. Wardlaw was not included as an author in the published article. The corrected Author Contributions and Funding statements appear below.JA performed the literature search, pooled analyses, statistical analysis and interpretation, manuscript drafting, and editing. LW performed pooling of databases, statistical interpretation, reviewed, and edited the manuscript. NS performed statistical interpretation, reviewed, and edited the manuscript. JW set up and co-ordinated all scan reading, including scan rating, training and data cleaning, reviewed and edited the manuscript. PB conceptualized the study, statistical interpretation, reviewed, and edited the manuscript, is corresponding author and has responsibility for submission. All authors contributed to the article and approved the submitted version.There was no specific funding for the present study. ENOS was funded by the UK Medical Research Council (G0501797) and BUPA foundation. RIGHT was funded by Nottingham University Hospitals NHS Trust (R&D Pump Priming Competition) and by the Division of Stroke, University of Nottingham. RIGHT-2 was funded by the British Heart Foundation (CS/14/4/30972). JW declares funding from the Scottish Imaging Network, A Platform for Scientific Excellence (SINAPSE) and the UK Dementia Research Institute which receives funding from the UK Medical Research Council, Alzheimer's Society and Alzheimer's Research UK. The funders were not involved in the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Natural Products and Bioprospecting. We deeply mourn Professor Zhou\u2019s loss and dedicate this note to him.Professor Zhou Jun, a world-renown natural products chemist, and resource botanist, passed away on March 27, 2020, at the age of 89 in Kunming, China. The natural products and plant resource research field lost a noble and outstanding leader when he died since he had contributed very much to the field in a variety of aspects. He was also one of the honorary editors-in-chief of Professor Zhou was born on February 5, 1932, in Dongtai, Jiangsu Province, China. He started to study at the National College of Pharmacy in Nanjing in 1948. He was then educated in East China University of Chemical Technology from 1954 to 1958. After he graduated, he joined to Kunming Institute of Botany, the Chinese Academy of Sciences.Iphigenia indica. He found the active component gastrodin in Gastrodia elata and successfully developed the drug for migraine and neurasthenia therapy.Professor Zhou was an active and accomplished scientist who worked on the national demands. He and his colleagues searched diosgenin, hecogenin, and tigogenin from domestic plants as new raw materials for the production of contraceptive drugs in China. He also developed the technique for the production of colchicine from Professor Zhou was one of the pioneers in the Chinese phytochemistry society. He focused his research on plant saponins and cyclopeptides for many years. He published over 400 peer-reviewed scientific papers during his career, pushing forward the frontiers and developments of phytochemistry in China.Through the years, Professor Zhou mentored numerous postdoctoral fellows, Ph. D., and graduate students pursuing careers in academia and various other industries. He was an excellent mentor, supportive and simultaneously firm. A long list of students and young scientists have, directly or indirectly, benefited from his expert knowledge and encouragement and followed in his footsteps. In this issue, some of them, as authors, are publishing articles dedicated to him.Professor Zhou will be remembered as a significant pillar within the scientific and phytochemistry communities worldwide\u2014we mourn his passing deeply.Prof. Dr. Liu Ji-KaiNatural Products & BioprospectingEditor-in-Chief of Prof. Dr. Luo Xiao-DongNatural Products & BioprospectingExecutive Editor-in-Chief of"} +{"text": "Previous studies have observed that physical activity (PA) levels tend to be lower in the U.S. population than in many other countries. Within the U.S., PA levels in children are lower in the South than in other regions. Cross-country and interregional differences in PA have not been studied in young children. n = 2008) from Finland (n = 370), Germany (n = 85), Sweden (n = 706), and the U.S. (n = 847). The U.S. sample was drawn from centers in Washington State, Colorado, and Georgia/Florida. Children wore accelerometers for 7 days, and the data were reduced to daily minutes of light-, moderate- (MPA), vigorous- (VPA), and moderate-to-vigorous- (MVPA) intensity PA and sedentary behavior. Multiple regression was used to compare children across countries and across regions in the U.S, adjusting for wear time, body mass index, and demographic characteristics. In an ongoing study of children at genetic risk for Type 1 diabetes, PA was measured by accelerometry in samples of 5-year-old children (p < 0.0001). Within the U.S., children's PA was consistently lowest in Georgia/Florida and highest in Washington. After adjusting for previously mentioned factors, MVPA and MPA were lower in U.S. children than those in Finland and Sweden. Estimates of physical activity were higher in Finland than in other countries, although not all comparisons were significantly different. U.S children spent significantly more time in sedentary behavior than children in Finland ( Cross-country differences in PA, previously reported for adults and adolescents, are evident in 5-year-old children. In general, PA levels are lower in U.S. children than their European counterparts, and within the U.S., are lower in Georgia/Florida and Colorado than in Washington. Future studies should be designed to identify the factors that explain these differences. Physical activity exerts a powerful and beneficial influence on a wide range of health outcomes , and theIncreases in the prevalence of obesity have been observed in most countries in the world, but those increases have been particularly dramatic in North America , 7. In tTEDDY, The Environmental Determinants of Diabetes in the Young, is an ongoing study seeking to identify the environmental triggers of type 1 diabetes (T1D) in genetically at-risk children . The stun = 183).A cross-sectional study design was applied using data from The Environmental Determinants of Diabetes in the Young (TEDDY) study. TEDDY is an ongoing prospective cohort study that is examining factors that relate to the development of T1D in a sample of genetically at-risk children from 4 countries . In the U.S., data are being collected from 3 regions, in Colorado, Georgia/Florida (Southeast), and Washington. The details of the TEDDY study design and methodology have been published previously , 16. Bri2) was calculated from the average height and weight for each participant.At the child's 9-month-old clinic visit, parents reported child demographic characteristics, including gender, race, ethnic minority status, and maternal education. In the U.S., race was reported as Hispanic, White Non-Hispanic, African-American Non-Hispanic, and Other. U.S. participants were classified as \u201cethnic minority\u201d if \u201cthe mother's first language was not English, the mother was not born in the U.S., or the child was identified as a member of an ethnic minority group based on the U.S. Census definition\u201d . For EurPhysical activity at the child's 5-year-old visit was measured by accelerometry. Participants were given an ActiGraph accelerometer to wear for at least 7 consecutive days. Monitors were worn on elastic belts around the waist, with the device placed on the right hip, during all waking hours, exclusive of water activities. The accelerometers were initialized to collect data at 80\u2009Hz and were downloaded as 1-second epoch data. Using SAS programs, data were reintegrated into 60-second epoch data files and analyzed for activity intensity levels. Periods of \u226560 minutes of consecutive zero counts were defined as nonwear and set to missing. Age-specific cut-points were used to determine the average minutes per day each child spent in light- (101-1290 counts per minute (cpm)), moderate- , vigorous- , and moderate-to-vigorous- (MVPA) intensity PA (>1291\u2009cpm), as well as total (light+MVPA) PA (TPA) and sedentary behavior , 20. Minn (percent) for categorical variables for each country and region of the U.S. For descriptive variables of interest, analysis of variance (ANOVA) or logistic regression was used to determine if there were differences between the four countries and between the three U.S. regions.Demographic variables were summarized (means (SD) for continuous variables and p values less than 0.05 were considered to be statistically significant.Mixed model regression analyses, adjusted for wear time, were used to determine the influence of demographic factors including age, gender, mother's education, ethnic minority, BMI, and season on estimates of light, MPA, VPA, MVPA, TPA, and sedentary behavior. Two-tailed Linear mixed regression models with least square means, which were Bonferroni corrected, were used to determine if intercountry differences existed for each outcome variable of interest: sedentary, light, MVPA, VPA, and TPA min/day, after adjusting first for monitor wear time, and demographic factors including age, gender, mother's education, ethnic minority, season, and BMI. Models were also fit to determine if there were differences in the outcome variables between the U.S. regions after adjustment for wear time, and demographic variables including age, gender, mother's education, race/ethnicity, season, and BMI. All analyses were performed in SAS (9.4).Participant characteristics are summarized in Results from the multiple linear regression analyses for PA and sedentary variables and demographic factors are presented in Comparisons across countries for the PA variables and sedentary behavior are summarized in The findings for comparisons across the three U.S. regions are also presented in The major finding of this study was that 5-year-old children in the U.S. were observed to be less physically active, in general, than those in Finland, Germany, and Sweden. Time spent in MVPA was significantly lower in U.S. children than in those from Finland and Sweden, and this was observed in both unadjusted analyses and after adjustment for demographic characteristics and weight status. To our knowledge, this is the first cross-country comparison of PA in children as young as 5 years, and it is one of very few such studies to have used accelerometry as an objective measure of PA. In a comparison of PA among 9-11-year-old children from 12 countries, the highest levels of activity were found in Finnish children and U.S. children were the least active based on data from accelerometers , 22. HowWe found that PA levels differed in children across the three U.S. regions from which the sample was drawn. Physical activity was highest among children in the Northwest region (state of Washington) and lowest in the Southeast region (Georgia/Florida). Children in the Central region (state of Colorado) were midway between children from the other two regions. Similar patterns were found for all expressions of physical activity , and both with and without adjustment for demographic factors and weight status. These findings are unique in that this is the first study to report regional differences in physical activity levels in U.S. children as young as 5 years of age. However, the regional pattern observed in this study is similar to patterns previously reported in studies of older children. For example, the Trial of Activity in Adolescent Girls (TAAG) was a large-scale multicenter study that used accelerometry to assess PA in groups of girls recruited through study centers in six geographically distributed states . SimilarIn addition to the differences in PA, our study found cross-country differences in the amount of sedentary behavior among the children. Minutes of sedentary behavior were highest among U.S. children and lowest in Finnish children. Within the U.S., the Southern states had the highest sedentary minutes per day, followed by Colorado, with Washington having the lowest (not significantly different). These differences are important because there is a growing body of evidence linking high levels of sedentary behavior to adverse health outcomes , 29. FewThe strengths of this study include the use of an objective measure of PA in a large, multinational sample. In addition, data were collected year-round in all the countries, reducing the potential influence of seasonality on physical activity behaviors. It should be noted that the sites that participated in this study do not represent the whole of the geographic regions referenced (Europe or regions of the U.S.). The use of accelerometry to measure PA may result in underestimates of activity since they do not measure water or cycling activities well. Additionally, the use of accelerometers requires analysis decisions to be made regarding compliance, wear time, and cut-points that influence outcomes. Another limitation is that participants in this study were at an elevated risk for development of T1D, although children were excluded from the analysis if they were antibody positive at or before the time of the 5-year-old measurement. It is nonetheless possible that parental knowledge of this risk led to changes in children's behaviors, including their PA.Previous studies have observed that adolescents and adults in the U.S. are less physically active than their counterparts in European countries, and the findings of the present study support the conclusion that this pattern extends to children as young as 5 years of age. Within the U.S., we observed that 5-year-old children in the Southeast region were less physically active, in general, than those in the West region. These differences persisted after adjustment for child-level characteristics, including gender, weight status, race/ethnicity, mother's education, and season of the year. Future studies should be designed to identify social and physical environmental factors and cultural characteristics that may explain why young children's PA levels differ across countries and across U.S. regions."} +{"text": "The following information is missing from the Funding statement: The authors declare that the project was funded by National Health Mission, J&K. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The Acknowledgements section for this paper is incorrect. The correct statement is: The authors are indebted to Mission director, National Health Mission, J&K for providing financial aid for this study. The authors also extend their gratitude to professor Samia Rashid, Principal/Dean Government Medical College Srinagar for her support. We also thank HOD Psychiatry, Professor Mohammad Maqbool Dar for his insightful guidance. Lastly, we thank COVID-19 survivors who participated in this study."} +{"text": "The aim of this study was to measure differences in quality-adjusted life expectancy (QALE) by income in Korea at the national and district levels.Mortality rates and EuroQol-5D (EQ-5D) scores were obtained from the National Health Information Database of the National Health Insurance Service and the Korea Community Health Survey, respectively. QALE and differences in QALE among income quintiles were calculated using combined 2008\u20132014 data for 245 districts in Korea. Correlation analyses were conducted to investigate the associations of neighborhood characteristics with QALE and income gaps therein.QALE showed a graded pattern of inequality according to income, and increased over time for all levels of income and in both sexes, except for low-income quintiles among women, resulting in a widened inequality in QALE among women. In all 245 districts, pro-rich inequalities in QALE were found in both men and women. Districts with higher QALE and smaller income gaps in QALE were concentrated in metropolitan areas, while districts with lower QALE and larger income gaps in QALE were found in rural areas. QALE and differences in QALE by income showed relatively close correlations with socioeconomic characteristics, but relatively weak correlations with health behaviors, except for smoking and indicators related to medical resources.This study provides evidence of income-based inequalities in health measured by QALE in all subnational areas in Korea. Furthermore, QALE and differences in QALE by income were closely associated with neighborhood-level socioeconomic characteristics. Socioeconomic inequalities in life expectancy (LE) have been well documented \u20133. HowevAt the national level, socioeconomic inequalities in health status are well established, whereas less is known about geographic variations in health status and socioeconomic health inequalities at the subnational level . The heaIn South Korea (hereafter \u2018Korea\u2019), the National Health Information Database (NHID) of the National Health Insurance Service and the In this study, we aimed to calculate the quality-adjusted life expectancy (QALE), which is an HE metric, according to income at the national and district levels, and to identify the relationships of neighborhood characteristics with QALE and income gaps therein.The study was approved by the National Health Insurance Service of Korea (NHIS-2018-1-430) and the Seoul National University Hospital Institutional Review Board (IRB No. E-1810-008-975).The information on mortality and HRQoL required for QALE calculations was obtained from the NHID and KCHS, respectively, according to gender, income, and district. Both the NHID and KCHS are considered to be good sources for monitoring income-based health outcomes at the district level, as they have district-level population representation and contain information on household income , 9. A toIncome was classified into five groups by calculating the quintile of the equivalized income considering the number of households by gender and age. District-level neighborhood characteristics included socioeconomic factors , health behaviors , and healthcare factors . A total of 11 components were used to construct the area deprivation index. Further details on these variables are presented in the The KCHS contains the EuroQOL five-dimensional (EQ-5D) 3-level questionnaire, which is a self-reported HRQoL tool that consists of five dimensions , each of which is scored with one of three levels of severity . The EQ-5D questionnaire profiles, which contain 243 possible health states, were matched to Korean population-based preference weights for EQ-5D , and theBased on the calculated mortality rates and the EQ-5D scores, LE and QALE were estimated using the Sullivan method. LE was estimated by calendar year, gender, and income level at the national and district levels during 2008\u20132014, and QALE was estimated by gender and income levels for 2008\u20132014 at the district level. The formula used for LE and QALE estimates can be found in the Table Figure\u00a0Figure\u00a0Figure\u00a0This study presented differences in HE by income, as measured by QALE, at both the national and district levels, and confirmed that income-related inequalities in HE existed in both men and women in all 245 districts of Korea. Geographic variations in health inequalities at the subnational level provide a distinct perspective on health inequalities in comparison to geographic variations in health . For example, in this study, the district with the highest HE (the Bundang district) presented an inter-quintile difference in HE of 6.2\u2009years (men and women combined). The HE in the lowest 20% income group of this district was equivalent to the average HE of the 21st district in the HE ranking. Within-district HE inequalities by income provide valuable information for district governments to plan policies and programs to reduce health inequalities in their own districts. Meanwhile, HE and differences in HE by income varied substantially across districts, and these differences were correlated with neighborhood-level socioeconomic characteristics.To the best of our knowledge, no prior study has examined geographic variations in income-related inequalities with respect to QALE at the subnational level. Chetty and colleagues examinedThe results of this study showed that pro-rich inequalities were more prominent in QALE than in LE. This held true for both men and women, for all years considered, and for all 245 districts . QALE did not increase in the low-income quintiles of women. A recent projection study indicated that Korean women\u2019s LE is expected to be the highest worldwide, with a 57% likelihood of it surpassing 90\u2009years in 2030 . The resThis study showed that the correlation between neighborhood characteristics and QALE was somewhat stronger in men than in women. Among the neighborhood characteristics, socioeconomic characteristics (the area deprivation index and Gini index) and current smoking prevalence showed stronger correlations with QALE than healthcare factors. A recent US study similarly showed that county-level LE had closer correlations with socioeconomic and race/ethnicity factors, as well as with behavioral and metabolic risk factors, than with health care factors . A priorThis study has certain limitations. First, when calculating QALE, the EQ-5D score of the 20- to 24-year-old age group was used for younger ages. This was because the KCHS surveys, which were the source of the EQ-5D data, were only conducted among those aged 19 and over. In practice, information on LE and HE at 0\u2009years old, which is a summary of the entire lifespan, is more useful for planning and evaluating health policies. Second, spatial autocorrelation was not considered in the analysis. This choice was made to allow local government officials and local health departments to utilize statistical findings from the data obtained from their own districts. Third, the magnitude of the relationships of LE and HE with income or district area should not be interpreted as causal effects of having more money or living in those specific districts. This study was conducted to describe the magnitude of these associations, rather than to explore causal effects.This study revealed the existence of income-based inequalities in health measured by QALE in all subnational areas in Korea, and showed close associations of the magnitude of QALE and income gaps therein with neighborhood socioeconomic characteristics.Additional file 1 Supplementary methods. Supplementary Table 1. Number of subjects from the National Health Information Database of National Health Insurance Service by year and income quintile. Supplementary Table 2. Number of deaths from the National Health Information Database of the National Health Insurance Service by year and income quintile. Supplementary Table 3. Number of subjects from the Korean Community Health Survey by year and income quintile. Supplementary Table 4. Central tendency (mean and median) and dispersion for district-level quality-adjusted life expectancy (QALE) and life expectancy (LE) by income quintile, 2008-2014. Supplementary Table 5. Correlations of the area deprivation index with district-level quality-adjusted life expectancy (QALE) by gender and income quintile, 2008-2014. Supplementary Table 6. Correlations of the area deprivation index with district-level life expectancy (LE) by gender and income quintile, 2008-2014. Supplementary Table 7. Correlations of district characteristics with district-level quality-adjusted life expectancy (QALE) by gender and income quintile, 2008-2014. Supplementary Table 8. Correlations of district characteristics with district-level life expectancy (LE) by gender and income quintile, 2008-2014. Supplementary Figure 1. Correlations of quality-adjusted life expectancy (QALE) with inter-quintile income differences in QALE at the district level. Supplementary Figure 2. Plots of correlations of the area deprivation index with district-level quality-adjusted life expectancy (QALE) by gender and urbanization level. Supplementary Figure 3. Plots of correlations of the area deprivation index with inter-quintile income differences in district-level quality-adjusted life expectancy (QALE) by gender and urbanization level. Supplementary Figure 4. Plots of correlations of district characteristics with district-level quality-adjusted life expectancy by gender and urbanization level. Supplementary Figure 5. Plots of correlations of district characteristics with inter-quintile income differences in district-level quality-adjusted life expectancy by gender and urbanization level.Additional file 2 : Supplementary Table 9. Quality-adjusted life expectancy (QALE) by income and gender among 245 districts in Korea, 2008-2014. Supplementary Table 10. Life expectancy (LE) by income and gender among 245 districts in Korea, 2008-2014. Supplementary Table 11. Distribution of population size and the number of death by income and gender among 245 districts in Korea, 2008-2014. Supplementary Table 12. Mean and 95% CI of EQ-5D scores by income and gender in Korea, 2008-2014. Supplementary Table 13. Distribution of EQ-5D scores by income and gender among 245 districts in Korea, 2008-2014."} +{"text": "Correction to: Stem Cell Res Ther 11, 320 (2020)https://doi.org/10.1186/s13287-020-01771-yAcknowledgement.Following publication of the original article , the autWe also want to acknowledge that this study was partially performed at the LCI facility/Nikon Center of Excellence, Karolinska Institutet, supported by grants from the Knut and Alice Wallenberg Foundation, Swedish Research Council, KI infrastructure, Centre for Innovative Medicine and Jonasson center at the Royal Institute of Technology."} +{"text": "Purpose: MR-guided Radiation Therapy (MRgRT) allows for high-precision radiotherapy under real-time MR visualization. This enables margin reduction and subsequent dose escalation which may lead to higher tumor control and less toxicity. The Unity MR-linac integrates a linear accelerator with a 1.5T diagnostic quality MRI and an online adaptive workflow. A prospective international registry was established to facilitate the evidence-based implementation of the Unity MR-linac into clinical practice, to systemically evaluate long-term outcomes, and to aid further technical development of MR-linac-based MRgRT.Methods and Results: In February 2019, the Multi-OutcoMe EvaluatioN of radiation Therapy Using the MR-linac study (MOMENTUM) started within the MR-linac Consortium. The MOMENTUM study is an international academic-industrial partnership between several hospitals and industry partner Elekta. All patients treated on the MR-linac are eligible for inclusion in MOMENTUM. For participants, we collect clinical patient data and technical patient data which is defined as information generated on the MR-linac during treatment. The data are captured, pseudonymized, and stored in an international registry at set time intervals up to two years after treatment. Patients can choose to provide patient-reported outcomes and consent to additional MRI scans acquired on the MR-linac. This registry will serve as a data platform that supports multicenter research investigating the MR-linac. Rules and regulations on data sharing, data access, and intellectual property rights are summarized in an academic-industrial collaboration agreement. Data access rules ensure secure data handling and research integrity for investigators and institutions. Separate data access rules exist for academic and industry partners. This study is registered at ClinicalTrials.gov with ID: NCT04075305 .Conclusion: The multi-institutional MOMENTUM study has been set up to collect clinical and technical patient data to advance technical development, and facilitate evidenced-based implementation of MR-linac technology with the ultimate purpose to improve tumor control, survival, and quality of life of patients with cancer. Radiotherapy is an important pillar in the multimodality treatment of cancer. Recently, MR-guided Radiation Therapy (MRgRT) has been introduced, enabling high-precision radiotherapy under real-time MRI visualization , 2. RealTechnical innovations in radiation oncology, such as the MR-linac, are typically received with great enthusiasm by radiation oncologists and physicists, who are keen to see new technologies implemented in routine practice. Evidence supporting these new radiotherapy technologies is generally scarce . HoweverIn line with the R-IDEAL framework, The Multi-OutcoMe EvaluatioN of radiation Therapy Using the MR-linac Study (The MOMENTUM study) was established. The goals of the MOMENTUM study are to aid and accelerate the development of anatomic and functional MRgRT and to enable systematic evaluation of clinical outcomes of patients. Ultimately, the MOMENTUM study aims to assess the effectiveness and safety of MRgRT (R-IDEAL stage 2a and 2b) as the paradigm is extended beyond conventional approaches, thereby facilitating the evidence-based introduction of the MR-linac into clinical practice. In addition, this registry is designed to serve as a data platform for future research investigating the MR-linac.In this article, we describe the MOMENTUM study, a clinical and technical patient registry, the governance structure and the handling of patient confidentiality.The MOMENTUM study is a complex registry, integrating clinical and technical patient data. The aim of the MOMENTUM study is to provide a data-infrastructure to:Collect routine-care data for the evaluation of short- and long-term feasibility, safety, effectiveness, and toxicity of treatments on the MR-linac. This also facilitates evaluation of (early) cost effectiveness of MR-linac treatments.Aggregate technical patient data to further develop MR-linac software algorithms that drive the online adaptive workflow aiming to maximize the benefits of MRgRT.Create a repository of anatomical and functional MR imaging data supporting Stage 0 of the R-IDEAL framework and aiming to further develop MRgRT.https://clinicaltrials.gov/ct2/show/NCT04075305). The MOMENTUM study was aided by a professional public-private partnership manager (Lygature).The MOMENTUM study was set up within the context of the international MR-linac Consortium, which currently consists of over 30 international centers . Four EuTwelve Tumor Site Groups (TSGs) were established within the Consortium: brain, bladder, breast, cervix, esophagus, liver, lung, oligometastases, oropharynx, pancreas, prostate, and rectal cancer, as seen in The MR-linac Consortium includes a Data Management Task Force (DMTF) which provides oversight and governance and manages the exchange of data according to the data access rules. The DMTF includes radiation oncologists, an Elekta representative, a physicist, and an epidemiologist.For each patient in the MOMENTUM study, a core set of clinical data items is collected, consisting of patient and tumor characteristics and outcome data. These outcome data include toxicity and cancer status data such as recurrence, disease-free, and overall survival . Within Fundamental to any radiation therapy database is the collection of technical patient data such as imaging information, radiotherapy structures (contours), plans, and dose information. The MOMENTUM study captures these data elements as well as others uniquely offered as a result of the novel, real-time MR-guided adaptive paradigm available on the MR-linac. Technical patient data, ultimately defined as data generated by the MR-linac during clinical operation, are captured after administration of radiotherapy on the MR-linac. Furthermore, researchers can collect extra research MRI scans during the treatment on the MR-linac of patients who consent to the optional research scans.At every institution, dedicated clinical research coordinators collect clinical and technical patient data from the hospital information system. The information is pseudonymized on site before the coordinators upload the data into the MOMENTUM database. Each patient is assigned a unique study ID (study identification number) which facilitates the link between the technical and clinical patient data. Furthermore, this study ID is used in any future studies on the MR-linac, if applicable.The study ID allocation is treatment-based, therefore the study ID relates to a radiation treatment course for individual patients. Multiple courses for a single patient are entered as separate events in the database but the IDs are linked facilitating the identification of a single patient with multiple treatment courses.The MOMENTUM clinical registry follows a centralized approach enabling secure virtual storage and easy uploading of data . The webThe technical database is hosted on a cloud computing platform and facilitates uploading of technical patient data, or DICOM data from all over the world. At every site, DICOM data are pseudonymized with the RSNA CTP by a dedicated technical patient data custodian on an on-premise workstation. After pseudonymization, technical patient data are uploaded to the cloud via a secure transfer using Microsoft Fast Data Transfer . Stored data are encrypted and accessible by researchers via secure VPN connection or virtual machines hosted on the cloud.The clinical and technical repositories function as two separate entities for which the data are only connected through the study ID.Within the MOMENTUM study, we aim for maximum clinical and technical patient data interoperability according to the previously mentioned FAIR principles . The linData quality is assured by using standardized electronic case report forms (eCRFs) and by the use of automatic validation and verification rules to reduce human error. Also, on-site training of the clinical research coordinators, standard operating procedures (SOPs), and data monitoring will ensure data quality. After one on-site monitoring visit the sites will be monitored through queries and questionnaires for data quality, database completeness, protocol compliance, and compliance to data protection legislation.The MOMENTUM study has been set up as an academic-industrial collaboration managed by the independent not-for-profit organization Lygature. This legal framework was selected because it best reflected the multi-pronged mission of clinical and technical development and clinical assessment. In addition, an academic-industrial partnership was felt to maximize transparency for academic and industrial collaborators.All patients over 18 years old treated on the MR-linac are eligible for participation in the MOMENTUM study. They provide informed consent for the collection of their pseudonymized clinical and technical patient data. Participants can consent to the collection of additional patient reported outcomes and additional MRI scans on the MR-linac. Furthermore, participants from the Netherlands can decide not to share their data with Elekta.The MOMENTUM study complies with national and regional data processing rules and regulations such as the European General Data Protection Regulation (GDPR), the USA Health Insurance Portability and Accountability Act (HIPAA), and the Canadian Personal Information Protection and Electronic Documents Act (PIPEDA). All rules and regulations on data sharing, data access, and intellectual property are summarized in an Academic Industrial Collaboration agreement (AIC) and signed by the MOMENTUM partners.As stated in the AIC, every institution that contributed to the MOMENTUM registry will have full control over the patient data they contributed. Therefore, the institutions function as their own data controllers. The UMC Utrecht will monitor registered data of all institutions separately throughout the duration of the study to ensure data quality and adherence to data protection legislation.Data access is regulated by data access rules which ensure secure, equitable, and safe data handling whilst allowing for open international data exchange in accordance with the FAIR principles. The data access rules are explicitly defined to facilitate research and encourage collaboration between MOMENTUM partners whilst safe-guarding the interests and rights of institutional representatives, TSG representatives, and primary investigators of future research performed on the MR-linac.For academic partners, we have developed a data request procedure for three different data request types . Data reData access by the industry partner Elekta, is governed through separate data access rules. Data requests from the industry partner are categorized into requests for technical patient, classifier, and outcome data . ClassifThe MOMENTUM study is financially supported by Elekta AB for 5 years and through in-kind contributions from all participating institutions. Conflicts of interest have been thoroughly addressed by recording academic and industry rights and obligations in the AIC, data access rules, and patient consent forms.Increased digitalization and technical developments have resulted in the release of promising technical innovations into the medical field. Unlike new pharmacological agents, these innovations and devices undergo limited comparative evaluation as they are approved for release onto the market based on only limited evidence of effectiveness. Acquiring robust data to prove the efficacy and added value of new innovations over standard treatment strategies is challenging and expensive. As a result, there is a lack of high level evidence that supports the use of new technologies in medicine, including in the field of radiation oncology .The MOMENTUM study has been designed to reverse this trend by providing an infrastructure for the evidence-based introduction of MR-linac technology, one of the most recent innovations in radiation oncology . Two uniIndustry-funded collaboration with academic institutions aiming to critically evaluate this new technology early in the process of clinical implementation.A well-integrated and governed technical and clinical database designed by radiation oncologists.Standardized data sets of prospectively accrued data for patients treated with this new technology.A methodology framework (R-IDEAL) which TSGs can adopt to demonstrate cancer specific applicability of the MR-linac treatments and ultimately compare MR-linac treatment to standard radiotherapy treatments or alternate treatment approaches.Easy and safe international data sharing between all partners, enhancing possibilities for collaborative studies.One of only a few industry-funded collaborations aiming to critically analyze the device right at the start of clinical implementation.A large scale academic-industrial partnership with all intellectual property rights and data sharing regulations recorded in a collaboration agreement (AIC). Therefore, facilitating industry sponsorship while maintaining an objective environment for academic partners to publish results and to enable Elekta to use pseudonymized technical patient data for further development of the MR-linac.The multi-institutional MOMENTUM study was set up to facilitate the evidenced-based implementation of the Elekta MR-linac technology and to support its further technical development. The aim of this new technology is to improve tumor control, survival, and quality of life of cancer patients treated with radiation therapy. The registry study was set up to facilitate the use of the R-IDEAL framework and evaluate the benefit of this technology. This study will facilitate high quality research in the field of radiation oncology.The study is approved by the Medical Ethics Committee (METC) Utrecht and the local Institutional Review Boards (IRB) of the participating institutions. The patients/participants provided their written informed consent to participate in this study.HV and WH contributed equally to this article. HV, WH, CF, and JC designed the study and are responsible for the general supervision of the study. SO, DE, EB, and HA have been involved in designing the study. SO drafted the manuscript and all authors critically revised the paper. All authors read, provided feedback, and approved the final manuscript.JC is Senior Vice President of Medical Affairs and Clinical Research Linac Based RT at Elekta. HA is senior Research Software Engineer at Elekta. KB is Distinguished Scientist at Elekta. AC receives research funding from Elekta. DE is Program Director, Clinical Registries at Elekta. BE receives institutional research and travel support from Elekta. CF-F receives research funding from Elekta. CF received funding and salary support related to the MR-Linac project from the National Institutes of Health (NIH), including: the National Institute for Dental and Craniofacial Research (NIDCR) Academic Industrial Partnership Grant (R01DE028290); NCI Early Phase Clinical Trials in Imaging and Image-Guided Interventions Program (1R01CA218148); an NIH/NCI Cancer Center Support Grant (CCSG) Pilot Research Program Award from the UT MD Anderson CCSG Radiation Oncology and Cancer Imaging Program (P30CA016672) and an NIH/NCI Head and Neck Specialized Programs of Research Excellence (SPORE) Developmental Research Program Award (P50 CA097007). CF received funding and salary support unrelated to this project from: the NIDCR Establishing Outcome Measures Award (1R01DE025248/R56DE025248); a National Institute of Biomedical Imaging and Bioengineering (NIBIB) Research Education Programs for Residents and Clinical Fellows Grant (R25EB025787-01); the NIH Big Data to Knowledge (BD2K) Program of the National Cancer Institute (NCI) Early Stage Development of Technologies in Biomedical Computing, Informatics, and Big Data Science Award (1R01CA214825); National Science Foundation (NSF), Division of Mathematical Sciences, Joint NIH/NSF Initiative on Quantitative Approaches to Biomedical Big Data (QuBBD) Grant (NSF 1557679); NSF Division of Civil, Mechanical, and Manufacturing Innovation (CMMI) standard grant (NSF 1933369). CF receives infrastructure support provided by the Multidisciplinary Stiefel Oropharyngeal Research Fund of the Charles and Daneen Stiefel Center for Head and Neck Cancer and the MD Anderson Program in Image-guided Cancer Therapy. CF has received direct industry grant support, honoraria, and travel funding from Elekta AB. JG was Senior Vice President Medical Affairs at Elekta at time of writing. SH reports non-financial support from Elekta , non-financial support from Merck Sharp and Dohme (MSD), personal fees, and non-financial support from Roche outside the submitted work. EH reports support from Cancer Research UK Programme Grant (A25351) and the National Institute for Health Research (NIHR) Biomedical Research Center at The Royal Marsden NHS Foundation Trust and the Institute of Cancer Research, London. The views expressed are those of the author(s) and not necessarily those of the NIHR or the Department of Health and Social Care. EH reports grants and non-financial support to host institution from Merck Sharp and Dohme, Astra Zeneca, and Bayer, and grants to host institution from Accuray, Varian, Janssen-Cilag, Kyowa Hakko UK, Alliance Pharma (previously Cambridge Laboratories), and Aventis Pharma Limited (Sanofi), outside the submitted work. KH reports support from Elekta MR-Linac Consortium, CRUK ART-NET Network Accelerator Award, CRUK Head, and Neck Programme Grant (C7224/A23275), ICR CRUK Center Grant, ICR/Imperial CRUK Convergence Center Grant and the Royal Marsden Hospital/ICR NIHR Biomedical Research Center. UH receives Elekta and Philips research funding. RH receives institutional funding from Elekta and institutional funding from MSD and ROCHE. AK reports support from Elekta MR-Linac Consortium, CRUK Programme Grant to Royal Marsden/Institute of Cancer Research (ICR) Radiotherapy Physics Department (A19727), and the Royal Marsden Hospital/ICR NIHR Biomedical Research Center. SL receives research funding, honoraria, and travel expenses from Elekta. MN receives research funding from Elekta. KO is program manager of the MOMENTUM project funded by Elekta AB, Stockholm, Sweden. AS is advisor/consultant with Abbvie, Merck, Roche, Varian , Elekta (Gamma Knife Icon), BrainLAB, and VieCure . Board Member: International Stereotactic Radiosurgery Society (ISRS). Past educational seminars with Elekta AB, Accuray Inc., Varian (CNS Teaching Faculty), BrainLAB, Medtronic Kyphon. Research grant with Elekta AB Travel accommodations/expenses by Elekta, Varian, BrainLAB. AS also belongs to the Elekta MR Linac Research Consortium, Elekta Spine, Oligometastases and Linac Based SRS Consortia. CS receives Institutional research support from Accuray, Siemens Healthineers, Elekta AB, Philips Healthcare, Manteia Technology and Honoraria and Travel funding from Elekta AB, Accuray. RT receives institutional research support from Elekta and Philips Honoraria (follow ms) and research grants from the Dutch Cancer Society, ZonMw, Health Holland, and NOW. AT received research funding and honoraria from Elekta and research funding from Varian and Accuray. WH receives institutional research and travel support from Elekta. HV receives research funding from Elekta. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The handling editor declared a past co-authorship with one of the authors CS."} +{"text": "The epidemiology of esophageal cancer (EC) can elucidate its causes and risk factors and help develop prevention strategies. We aimed to provide an overview of the burden, trends, and risk factors of EC in China from 1990 to 2017. We also investigated the differences between China, Japan, and South Korea and discussed the possible causes of the disparities.We used the Global Burden of Disease Study 2017 to obtain data on incident cases, deaths, disability-adjusted life-year (DALY) cases, age-standardized incidence rate (ASIR), age-standardized death rate (ASDR), and age-standardized DALY rate of EC in China, Japan, and South Korea from 1990 to 2017. Trend analysis was performed using joinpoint analysis. We measured the associations between ASIR, ASDR, and age-standardized DALY rate and the socio-demographic index (SDI) for 1990\u20132017. We also analyzed the risk factors associated with EC deaths and DALYs.China recorded 234,624 incident cases of and 212,586 deaths from EC in 2017. The ASIR and ASDR declined from 1990 to 2017. Until 2017, the ASIR was 12.23, and ASDR was 11.25 per 100,000 persons. The DALYs were 4,464,980 with an age-standardized rate of 222.58 per 100,000 persons in 2017. The ASIR, ASDR, and age-standardized DALY rate in China were twice those of Japan and South Korea. These three indicators showed a decreasing trend, whereas SDI increased, in all three countries from 1990 to 2017. Tobacco and alcohol use remained the major risk factors for EC death and DALYs, especially for men in China and women in Japan and South Korea. High body mass index (BMI) and low-fruit diet were the main risk factors for women in China.The incident cases and deaths of EC in China, Japan, and South Korea increased from 1990 to 2017, whereas the ASIR, ASDR, and age-standardized DALY rate declined. China had the greatest burden of EC among three countries. SDI and aging along with tobacco use, alcohol use, high BMI, and low-fruit diet were the main risk factors of death and DALYs and should be paid more attention. Esophageal cancer (EC), the seventh most common cancer and the sixth leading cause of cancer death worldwide , etiologThe current study aimed to provide an up-to-date overview of the burden, trends, and risk factors of EC in China from 1990 to 2017, compared with Japan and South Korea, and to discuss the potential reasons for the disparities. In considering the availability and comparability of data, we conducted our analysis using data from the Global Burden of Disease Study 2017. We investigated the factors for EC, including the socio-demographic index (SDI), age, sex, and various risk exposures, to contribute to strategies for health promotion.https://ghdx.healthdata.org/gbd-results-tool). In the case of China, we analyzed data on a total of 34 province-level administrative units, including 22 provinces, four municipalities, five autonomous regions, Taiwan, Hong Kong Special Administrative Region (SAR), and Macao SAR. All of these are referred to as provinces throughout the present paper. According to the division of the National Bureau of Statistics of China, we classified the 31 provinces in mainland China into three areas according to geography: Eastern (n\u2009=\u200911), Middle (n\u2009=\u20098), and Western (n\u2009=\u200912). The DALYs and age-standardized DALY rate in 34 provinces of China from 1990 to 2017 were collected from a previous study [We obtained data on incident cases, deaths, disability-adjusted life-years (DALYs), age-standardized incidence rate (ASIR), age-standardized death rate (ASDR), and age-standardized DALY rate of EC in the world, East Asia and Pacific region, China, Japan, and South Korea for the period of 1990\u20132017 from the Global Burden of Disease Study 2017 , 16. TheWe performed joinpoint regression analysis to determine the annual percent changes in the ASIR, ASDR, and age-standardized DALY rate of EC from 1990 to 2017 in the world, East Asia and Pacific, China, Japan, and South Korea using Joinpoint (version 4.7.0). Joinpoint regression analysis uses a piecewise linear regression method to determine the trend displayed by one or more line segments .The exposure variables were divided into four large categories: environmental and occupational, behavioral, metabolic, and dietary risks. These risks are defined and described in the comparative risk assessment framework of the Global Burden of Disease Study 2017 .p value of\u2009<\u20090.05 was considered statistically significant.The standardized methods of the Global Burden of Disease Study 2017 have been reported .\u00a0To elimTable We observed an increasing trend in incident cases and deaths in the world, East Asia and Pacific, China, Japan, and South Korea for both sexes between 1990 and 2017 of the incident cases occurred in men, compared with 29.97% in women worldwide of the incident cases occurred in men, compared with 30.75% in women in China . Age-specific rates for both incidence and death increased with age; the same trend was seen in the world, East Asia and Pacific, and the three countries . The burden of EC was rare in young people, and both death and incident rates peaked at the age of\u2009\u2265\u200980\u00a0years in both sexes.p\u2009<\u20090.05, Fig.\u00a0p\u2009<\u20090.05; 2007\u20132013: by 2.97%, p\u2009<\u20090.05). Similarly, the ASDR in China declined in four periods .The results of the joinpoint regression analyses are shown in Fig.\u00a0p\u2009<\u20090.05, Fig.\u00a0p\u2009<\u20090.05; 2011\u20132015: declined by 2.30%, p\u2009<\u20090.05). The first increasing and then decreasing trend in ASDR was found between 1990 and 2017 .The trends in ASIR and ASDR in South Korea were similar to the trends in Japan . The trends in age-standardized DALY rate in China, Japan, and South Korea are displayed in Additional file 2: Figure S2.Meanwhile, Japan showed different trends in ASIR and ASDR. ASIR first increased in one period and then declined in two periods than their rural counterparts . Further, middle areas presented the highest ASIR and ASDR . The lowest ASIR and ASDR were reported in eastern areas .Table Generally, tobacco was the greatest contributor to death and DALYs in the world, East Asia and Pacific, China, Japan, and South Korea . We found differences in trends in the percentages of death and DALYs from tobacco use in the three countries from 1990 to 2017. Worldwide tobacco-related DALYs and death remained stable between 1990 and 2017. However, China showed markedly increasing trends in tobacco-related DALYs and death, whereas Japan exhibited markedly declining trends, and South Korea showed an increasing trend and then a decreasing one. Moreover, the percentage of high BMI-related DALYs and death increased and that of low-fruit diet decreased in these regions over time.The percentage of EC age-standardized deaths (ASDs) owing to risk factors differed across countries and sexes in 2017 Fig.\u00a0a, b. SpeIn 2017, the percentage of ASDs owing to tobacco and low-fruit diet for women was the highest in China and the lowest in South Korea . South Korea (21.2%) and Japan (21.0%) had the highest percentage of ASDs owing to alcohol use for women, exceeding the level of East Asia and Pacific (6.1%) and the global level (9.7%); China had the lowest percentage (5.2%). As for the percentage of ASDs owing to high BMI for women, the lowest was found in Japan (11.7%) and the highest in China (16.6%). In summary, tobacco and alcohol use were the most important risk factors for ASDs in men in three countries and in women in Japan and South Korea. Meanwhile, in Chinese women, high BMI and low-fruit diet were the risk factors that contributed most to ASDs.The percentages of EC age-standardized DALY rate owing to risk factors also differed across countries and gender sexes in 2017Fig.\u00a0a, b. TobThe burden and trends of EC in the world, East Asia and Pacific, China, Japan, and South Korea between 1990 and 2017 were systemically described in the present study. The incident and death cases of EC in the world, East Asia and Pacific, and the three countries increased from 1990 to 2017, whereas ASIR, ASDR, and age-standardized DALY rate decreased. In addition, we found that the incident and death rates of EC increased with age, especially in people aged\u2009\u2265\u200945\u00a0years, peaking at the seventh and eighth decades of life in the world, East Asia and Pacific, China, Japan, and South Korea. As populations age worldwide, EC will create a heavier burden on public health care systems.Our results confirmed that men carried the majority of the incident and death rates of EC , 21. HowThe regional variations in the ASIR and ASDR of EC for both sexes are huge across China, Japan, and South Korea, which may be explained by the heterogeneity in the prevalence of risk factors. Socioeconomic status is a risk factor for EC. In our study, we used SDI to reflect socioeconomic status. All ASRs decreased as the SDI increased. However, the association between socioeconomic status (SDI) and the ASIR, ASDR, and age-standardized DALY rate is complex and nonlinear. The declines we found might be from increasing socioeconomic levels and dietary improvements. Although the ASIR and ASDR in China declined with SDI value, they were above the expected levels in the past 28\u00a0years and may be attributed to the behavioral and dietary risk factors of EC and lack of early diagnosis and monitoring methods. Meanwhile, the ASIR in South Korea was much lower than the expected levels, which, we speculate, can be attributed to the prevention of the other risk factors of EC in South Korea.2 enhances the risk of EC by approximately 10% [Our study revealed that China, Japan, and South Korea had different risk factor profiles of EC. A 2005 report attributed 46% of the incidence of and death from EC in China to smoking, alcohol drinking, and low intake of vegetables and fruits . Tobaccotely 10% . A systetely 10% . Our stutely 10% . In all,Helicobacter pylori\u00a0infection, should be investigated in future studies.Our study has some limitations. First, the study lacked information on the histological subtypes of EC. The epidemiological features of ESCC and EAC appear to differ substantially. However, owing to a lack of necessary data, we could not identify the burden and risk factors in these two histological subtypes. Second, data on the type of risk factors in this study were limited. More factors that might be attributable to EC, such as Incident cases and deaths in China, Japan, and South Korea increased from 1990 to 2017, whereas the ASIR, ASDR, and age-standardized DALY rate declined. With an aging population, EC will create a heavier burden on public health care systems. Risk factors, including smoking, alcohol use, high BMI, and low-fruit diet, are the main factors of death and DALYs and should thus be given importance.Additional file 1:Figure S1. Age-specific rates for incidence, death, and DALY increased with age. DALY: disability-adjusted life-year.Additional file 2:Figure S2. Trends for age-standardized DALY rate of esophageal cancer in China (A), Japan (B), South Korea (C), East Asia and Pacific (D), and the world (E) from 1990 to 2017 calculated by joinpoint regression analyses. DALY: disability-adjusted life-year.Additional file 3:Figure S3. Trends in percentage of esophageal cancer all-age death and ASDR due to risk factors in the world, East Asia and Pacific, China, Japan, and South Korea, from 1990 to 2017. (A) all-age death; (B) ASDR. ASDR: age-standardized death rate.Additional file 4:Figure S4. Trends in percentage of esophageal cancer all-age and age-standardized DALY rate due to risk factors in the world, East Asia and Pacific, China, Japan, and South Korea, from 1990 to 2017. (A) all-age DALYs; (B) age-standardized DALY rate. DALY: disability-adjusted life-year.Additional file 5:\u00a0Table S1. Incident cases, death cases, age-standardized incidence rate (ASIR), and age-standardized death rate (ASDR) for esophageal cancer in China, by geographic areas, 2015."} +{"text": "Journal of Patient-Reported Outcomes (JPRO) is an international, open access, multi-disciplinary journal publishing original research and review articles, brief communications, commentaries, editorials, and reviews of recent books and software advances in the field of patient-reported outcomes. The JPRO is an official journal of the International Society for Quality of Life Research (ISOQOL). The first JPRO articles were published on September 12, 2017.The Quality of Life Research\u201d and was the \u201cbeginning of a beautiful journal\u201d (https://jpro.springeropen.com/articles/10.1186/s41687-017-0009-2). The success of the JPRO is due to their extensive knowledge, hard work and dedication. The journal\u2019s success also reflects the diligent contributions made by reviewers and the quality of the science in the published manuscripts.Dennis Revicki and David Feeny were the founding Co-Editors-in-Chief. Dennis served until the start of 2020 and David finished at the end of 2020. They noted at the onset of the JPRO that it was \u201cintended to complement and extend ISOQOL\u2019s existing journal, Quality of Life Research (2009\u20132017) and David served as ISOQOL president (2004\u20132005). We are grateful for the giant steps forward they made with the JPRO.Dennis and David are luminaries in the field of patient-reported outcomes. Both are former ISOQOL President\u2019s Award recipients (Dennis in 2007 and David in 2010). In addition, Dennis is a former co-Editor-in-Chief of We hope to continue the upward trajectory of the journal and obtain the coveted impact factor soon. We encourage those doing patient-reported outcomes research to submit manuscripts and to serve as peer reviewers for the JPRO. We can only move forward with your help.Sincerely,Ron D. Hays and Chih-Hung ChangJPRO Co-Editors-in-Chief"} +{"text": "Correction to: BMC Public Health 20, 13 (2020)https://doi.org/10.1186/s12889-019-8123-0It was highlighted that in the original article referencStatistical analysisHierarchical logistic regression , which accounted for the clustering at the census block group level, was used to calculate odds ratios (OR) of multimorbidity for the highest vs. lowest quintile of each individual measure in the ADI. An unadjusted model, a model with adjustment for age , sex, race , and ethnicity , and a fully-adjusted model with further adjustment for individual level of education were run. Hierarchical logistic regression was also used to model the association of the composite ADI with multimorbidity. Unadjusted and multivariable adjusted models (as defined above) were run. The models were repeated for severe multimorbidity (\u22655 chronic conditions). In addition, we tested 2-way interactions between ADI and age, between ADI and sex, and between ADI and individual level of education. Forest plots were used to display the fully-adjusted ORs in graphical form in strata by age (for the age by ADI interaction), by sex (for the sex by ADI interaction), and by education (for the education by ADI interaction)."} +{"text": "Accurate tourist flow prediction is key to ensuring the normal operation of popular scenic spots. However, one single model cannot effectively grasp the characteristics of the data and make accurate predictions because of the strong nonlinear characteristics of daily tourist flow data. Accordingly, this study predicts daily tourist flow in Huangshan Scenic Spot in China. A prediction method (GA-CNN-LSTM) which combines convolutional neural network (CNN) and long-short-term memory network (LSTM) and optimized by genetic algorithm (GA) is established. First, network search data, meteorological data, and other data are constructed into continuous feature maps. Then, feature vectors are extracted by convolutional neural network (CNN). Finally, the feature vectors are input into long-short-term memory network (LSTM) in time series for prediction. Moreover, GA is used to scientifically select the number of neurons in the CNN-LSTM model. Data is preprocessed and normalized before prediction. The accuracy of GA-CNN-LSTM is evaluated using mean absolute percentage error (MAPE), mean absolute error (MAE), Pearson correlation coefficient and index of agreement (IA). For a fair comparison, GA-CNN-LSTM model is compared with CNN-LSTM, LSTM, CNN and the back propagation neural network (BP). The experimental results show that GA-CNN-LSTM model is approximately 8.22% higher than CNN-LSTM on the performance of MAPE. The deepening of the reform and opening and the rapid development of the national economy has simultaneously seen the improvement of the economic capacity and living standard of the Chinese people. An increasing number of Chinese people now focus on better quality of life and higher levels of spiritual pursuit. The recent years witness the government\u2019s development of tourism. The development of the national tourism industry has been substantially promoted with various policies and activities. According to the 2018 report published by the National Tourist Bureau of China on the development of culture and tourism in 2018, the domestic tourism market maintained its steady growth; inbound tourism market grew slowly and steadily, whereas the outbound tourism market developed rapidly. Moreover, the total number of domestic tourists was 5.539 billion, inbound tourists was 141.2 million, outbound tourists was 149.72 million, and the total tourism revenue was 5.97 trillion yuan. In 2018, the total number of tourists was 6.024 billion of the 11,924 A-level scenic spots in China. The total tourism revenue of all A-level scenic spots had an increase of 7.8% over the previous year . This study uses the min-max normalization method, and the formula is shown below:This article selects daily historical data of Huangshan Scenic Spot from 2015 to 2018 as the original data. Then, data from 2015 to 2017 are used as the training set; data from 2018 are used as the test set.The experiments in this article are performed in the following hardware environment: CPU: Intel i5 9400f, Memory: 16GB, GPU: 1660ti. The software framework is a Tensorflow framework based on Keras deep learning tools, which is written by Python. Keras provides a simple and consistent programming interface that can help users quickly understand the neural network architecture and reduce the repetitive work in the code implementation process. Keras has the characteristics of modularity, and supports the free combination of model layers and layer-by-layer overlay.The constructed data set is divided into training and test sets, and the training set data are input into the model for training. The three-layer convolutional and pooling layers are selected according to the size of the unit feature map and the principle of the convolutional neural network. In the convolutional layer of the CNN module, the size of the unit feature map is 16 \u00d7 16, the size of the convolution kernel is set to 2, and the step size of the convolution kernel is set to 1. The pooling layer sets the same parameters as the convolutional layer. Then, weight of CNN is initialized. Meanwhile, CNN module also adds Dropout to reduce the probability of overfitting. The activation function is set to Selu function . Comparei is input value; yi is the output value after BN; m is the size of the mini-batch, that is, a mini-batch with m inputs; i; Some problems in deep neural network training are identified. For example, owing to the large number of layers in a deep neural network, changes occurring in the parameters of one layer will affect the output of all subsequent layers and result in frequent parameter modification and low training efficiency. In addition, before the data pass through the activation function, the output value of the nerve cell may also cause the failure of the latter to work if it remarkably exceeds the appropriate range of the activation function itself. Batch Normalization (BN) is desigIn the LSTM module, the weight is initialized using MSE as the loss function and Adam function as the optimizer, learning rate = 0.001, beta_1 = 0.9, beta_2 = 0.999. Here, our objective is to minimize the forecasting error of the model:To keep the impartiality of performance evaluation, only the training data is used during the training, while the testing data is not used. Each time the training data are input to the GA-CNN-LSTM, a loss value is generated, according to which the optimizer uses a backpropagation method to adjust the parameters of GA-CNN-LSTM. The forecast result of GA-CNN-LSTM will be more and more accurate with the increase of training iterations. After the GA-CNN-LSTM training is finished, the testing data is input into the GA-CNN-LSTM, and the testing results and real results are compared to evaluate the performance of the GA-CNN-LSTM.When there is not enough training data or when there is overtraining, overfitting may occur. However, there are many ways to avoid overfitting, such as regularization, data augmentation, dropout, dropconnect, or early stopping. The method used in this paper is dropout. CNN and LSTM module both add dropout to reduce the probability of overfitting. SDs of prediction result and the number of tourists. This study uses CNN-LSTM, CNN, LSTM, and BP as comparative experiments. The results of CNN-LSTM, CNN, LSTM, and BP will be compared with the experimental results of GA-CNN-LSTM. In addition, mean absolute percentage error (MAPE), root mean squared error (RMSE), Pearson correlation coefficient (r), Kling\u2013Gupta efficiency (KGE) and index of agreement (IA) are used as the criteria for measuring the pros and cons of the model, with MAPE as the main evaluation criterion:First, four CNN-LSTMs are manually selected with different numbers of neurons. avg= 20.77. For 4 different CNN-LSTMs, MAPEavg = 23.08, 22.97, 22.89, 22.62, respectively. From the experimental results, the performance of GA-CNN-LSTM on MAPE can be concluded to be better than CNN-LSTM.GA-CNN-LSTM, CNN-LSTM with the best performance, CNN, LSTM, and BP are selected as comparison experiments. On the performance of KGE in Test2, the results of the five algorithms from small to large is CNN (0.695), GA-CNN-LSTM (0.767), CNN-LSTM (0.772), LSTM (0.839), BP (0.872). KGE have an ideal value of 1. Therefore, from the perspective of KGE, GA-CNN-LSTM performs poorly. This is where improvement is needed. On the performance of RMSE in Test2, the results of the five algorithms from small to large is GA-CNN-LSTM (2983.33), CNN-LSTM (3185.16), CNN (3290.35), BP (3296.08), LSTM (3423.17). This proves that GA-CNN-LSTM is more accuracy.On the predictions for the second and third seasons, although GA-CNN-LSTM is not the best, its performance still belongs to the acceptable range, and the gap among the best algorithms is small. For annual performance, GA-CNN-LSTM shows better stability than the others.All these experimental results reflect the reliability and efficiency of GA-CNN-LSTM in tourist flow prediction. Although the prediction performance of both CNN and LSTM is good, GA-CNN-LSTM is better. It further proves that the method of extracting data features through CNN and predicting through LSTM is reliable.Tourism has slowly become an important part of the local and national economy. How to manage scenic spot scientifically and efficiently is an urgent problem for the scenic spot management department. The prediction of tourist flow is the premise of management. Only under the premise of accurate prediction can the scenic spot management department make a reasonable allocation of scenic spot resources and ensure the sustainable development of the scenic spot. This study takes the famous Huangshan Scenic Spot as an example. It uses environmental, historical data, and Baidu search index to construct a new data set to express tourist flow and establishes a GA-CNN-LSTM-based prediction method. At the same time, considering the lag period between web search and travel, through the correlation analysis, the Baidu search index with the most relevant lag periods between keywords and the total number of tourist flow are selected. Compared with other algorithms, this method predicts daily tourist flow more accurately than the other intelligent algorithms in MAPE, r and IA. However, some limitations are identified in the experiment, which are worthy of further research. Examples include how to select influencing factors, pre-process data, and construct convolutional neural networks, and so on. Although the accuracy of GA-CNN-LSTM is higher than that of other algorithms during the peak time, the overall prediction accuracy of the peak time remains insufficient. In general, the GA-CNN-LSTM prediction method proposed in this study provides new ideas for daily tourist flow prediction. This method has a good prospect in tourism management research and application and can establish a healthy tourism industry and sustainable development."} +{"text": "Pediatric emergency medicine (PEM) is a relatively recent subspecialty, recognized in the United States in 1992.Regardless of the maturity of PEM systems in developing countries, there is widespread disparity in mortality by geography and income. The burden of deaths of children in regions of the world such as sub-Saharan Africa and South Asia4The International Federation of Emergency Medicine (IFEM) is the umbrella association of EM globally, and is composed of over 60 national EM associations.Standards for the Care of Children in the Emergency Department (StandardsV3).StandardsV3 document (https://www.ifem.cc/wp-content/uploads/2019/06/Standards-of-Care-for-Children-in-Emergency-Departments-V3-2019.pdf).6As part of its effort to support and improve pediatric care globally, PEMSIG developed the third revision of the StandardsV3 are not the ultimate and most comprehensive guideline for pediatric emergency care. Rather, they form the foundation by providing recommendations and standards for any clinician and service that cares for children. Our hope is that the promulgation and dissemination of these StandardsV3 will augment clinical knowledge and basic equipment requirements, but also aid clinicians, managers, and policy-makers to advocate for improvements in the quality of emergency care of children. This in turn will promote more formal development of PEM systems at local and national levels.It is important to understand that these"} +{"text": "We report the clinical features of novel coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in a family setting of 13 people with person-to-person transmission in Yancheng, Jiangsu Province, China. Since the first case of a novel coronavirus disease 2019 (COVID-19) was detected in early December 2019, it has spread rapidly all over the world \u20136. In prData were collected from Yancheng Third People's Hospital of Jiangsu Province, China. A total of 13 patients from a family cluster were tested SARS-CoV-2 positive after seven of the family members had been to Wuhan. Patients were hospitalized from January 26, 2020 to February 28, 2020. Throat swab samples were collected, and SARS-CoV-2 was detected using qRT-PCR assay. CT and hematological examinations were performed. Patients were carefully monitored and treated during hospital isolation. This study was approved by the ethics committee of Yancheng Third People's Hospital of Jiangsu Province, and written informed consent was obtained. This study followed the reporting guideline for case series.As shown in We reported a cluster of 13 family members of infected with SARS-CoV-2. The uniqueness of this cluster is that only four people were infected during the wedding with so many people attending the wedding. Therefore, it has been assumed that infection of this virus is correlated with the strength of individual immunity , 8. FurtThe raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Yancheng Third People's Hospital. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin. Written informed consent was obtained from the individual(s), and minor(s)' legal guardian/next of kin, for the publication of any potentially identifiable images or data included in this article.HZ, JC, RC, and BC: data collection and interpretation. RC and HZ: original draft preparation. RC and BC: review and editing. BC, RC, and JC: supervision. All authors: reviewed and approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Nature Communications 10.1038/s41467-020-16902-5, published online 23 June 2020.Correction to: The original version of this Article contained an error in the author affiliation.Guy S. Salvesen was incorrectly associated with Department of Pharmacology; Vagelos College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "Nature Communications 10.1038/s41467-020-18269-z, published online 8 September 2020.Correction to: The original version of this Article contained errors in the author affiliations. J. M. Fink was incorrectly associated with \u2018Institute for Quantum Science and Technology (IQST), University of Calgary, Calgary, AB, Canada\u2019 in the HTML version of the paper.Additionally, the present address of the author S. Barzanjeh, \u2018Institute for Quantum Science and Technology (IQST), University of Calgary, Calgary, AB, Canada\u2019 was incorrectly assigned as a full affiliation in the HTML version of the paper.These errors have been corrected in the HTML version of the Article. The PDF was correct at the time of publication."} +{"text": "We are pleased to announce that International Brazilian Journal of Urology became the official communication organ of the American Confederation of Urology. The editorial board and the Brazilian Urology Society are very happy because the beginning of this partnership what will contribute to reinforce the impact of the International Brazilian Journal of Urology on the American continent.We are experiencing very difficult times because of the Covid-19 pandemic and at the time of writing this editorial a second wave of the disease is starting in Europe. However, research cannot stop. The March-April 2021 number of Int Braz J Urol, the ninth under my supervision, presents original contributions with a lot of interesting papers in different fields: Prostate Cancer, Urethral Stricture, Male Incontinence, Vesicoureteral Reflux, Renal Cell Carcinoma, Bladder Cancer, BPH, SARS-CoV-2 and Urology, Radiation Induced Cystitis, Vaginoplasty, Varicocele, Basic Research applied to Female Urinary Incontinence, Flexible Ureteroscopy, Penile Fracture and Straghorn Renal Calculi. The papers came from many different countries such as Brazil, USA, Turkey, China, Iran, Portugal, Serbia, Montenegro and Spain, and as usual the editor's comment highlights some of them.In the present issue we present four important papers in reconstructive urology and 2 very important papers about SARS-CoV-2 pandemic in Urology. Dr. Benson and colleagues from USA performed in page 237 a nice sDr. Angulo and colleagues from Spain and Portugal present The 2 papers about COVID-19 in urology came from Brazil. Dr. Mazzuchi and collegues from Brazil presented in page 251 an imporDr. Jeremias and collegues from Brazil on page Dr. Castellanti de Mattos and collegues from Brazil performeDr Menezes and collegues from BraDr. Wang and collegues from ChiDr. Loftus and collegues from USADr. Maluf and collegues from BraDrs. Djordjevic and Vokovic from SerWe hope that readers will enjoy the present number of the International Brazilian Journal of Urology in this very difficult times of COVID-19."} +{"text": "Nature Communications 10.1038/s41467-020-16318-1, published online 22 May 2020.Correction to: The original version of this Article omitted from the author list the 11th author Beisha Tang, who is from the \u2018Department of Neurology & Key Laboratory of Hunan Province in Neurodegenerative Disorders, Xiangya Hospital, Central South University\u2019. Consequently, the following was added to the Author Contributions: \u2018B.T. sponsored H.Y. to do research at Emory University and provided advice on the experiments\u2019. This has been corrected in both the PDF and HTML versions of the Article.In addition, the original version of this Article contained an error in the author affiliations. Yongcheng Pan was incorrectly associated with Department of Neurology, The First Affiliated Hospital, Sun Yat-sen University, 510080 Guangzhou, China and their affiliation with Department of Neurology & Key Laboratory of Hunan Province in Neurodegenerative Disorders, Xiangya Hospital, Central South University was inadvertently omitted. This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "JSR.Introducing two new Main Editors of Journal of Synchrotron Radiation.Dibyendu Bhattacharyya of Bhabha Atomic Research Centre and Kristina Kvashnina of the Rossendorf Beamline \u2013 The European Synchrotron have recently been appointed as Main Editors of Dibyendu Bhattacharyya completed his undergraduation and post-graduation studies at University of Calcultta, Kolkata, India, and obtained his PhD degree from Jadavpur University, Kolkata, in 1995 based on his work at the Indian Association for the Cultivation of Science, Kolkata. After spending a few years at Newcastle Photovoltaics Applications Centre, University of Northumbria, Newcastle Upon Tyne, UK, as post-doctoral researcher, Dr Bhattacharyya joined Bhabha Atomic Research Centre, Mumbai, India, as Scientific Officer in 1997 where he is presently heading the Synchrotron Science and Multilayer Physics Section of the Atomic and Molecular Physics Division. Trained as a thin film multilayer device researcher, Dr Bhattacharyya moved to the area of synchrotron research in 2004, when the construction of India\u2019s first hard X-ray synchrotron source Indus-2 began, and took a leading role in building up the two X-ray absorption spectroscopy beamlines. Over the years, Dr Bhattacharyya built up a highly professional group consisting of scientists, engineers and students which led to a very successful utilisation of these facilities leading to a large number of publications in reputed international journals and also contributing to product-oriented industrial research. Dr Bhattacharyya has over 260 publications in international journals and three book chapters to his credit with more than 4500 citations. He is recipient of several awards including the Homi Bhabha Scientific and Technical Excellence Award in 2014 and DAE-SRC Outstanding Investigator Award in 2012. Dr Bhattacharyya also holds the position of Professor in the Physical Sciences at Homi Bhabha National Institute, Mumbai, and has guided several PhD students.Kristina Kvashnina is ERC Group Head at the Rossendorf Beamline (BM20) of the European Synchrotron (ESRF), Grenoble, France, supported by Helmholtz-Zentrum Dresden-Rossendorf, Germany, and also Professor at the Department of Chemistry, Moscow State University, Russia. She completed her undergraduate degree in theoretical physics at Ural State University in 2000 and her PhD degree in experimental physics at Uppsala University in 2006, while carrying out experiments at the Advanced Light Source of Lawrence Berkeley National Laboratory, USA. She used soft X-ray absorption (XAS) and X-ray emission spectroscopy (XES) methods together with resonant inelastic X-ray scattering (RIXS) to study rare-earth containing materials. She then moved to the ESRF, where she conducted pioneering research on actinide systems using advanced high-energy-resolution X-ray spectroscopy methods (HERFD-XAS and RIXS) at the actinide M4,5 edges. In 2017, Kristina received a European Research Council (ERC) Grant to support her research on actinide and lanthanide nanomaterials aiming to better understand f-electron systems at the atomic level. In 2019 she received a Megagrant from the Ministry of Science and Higher Education, Russia. Kristina has published more than 140 papers in the area of X-ray spectroscopy, cited more than 2200 times, and has served as key note speaker, chair and co-chair, organizer and program committee member at numerous conferences in her area of expertise. Kristina is a member of several advisory groups and project review panels in Germany, Italy, UK, Russia, Switzerland and Hungary.Journal of Synchrotron Radiation since 2019, Dibyendu is a new addition to the editorial board. The two existing Main Editors (Yoshi\u00adyuki Amemiya and Ingolf Lindau) and Editor-in-Chief of IUCr Journals (Andrew J. Allen) are delighted to welcome Kristina and Dibyendu to the team, and look forward to working with them to further develop the journal for the synchrotron radiation and free-electron laser communities. They would also like to thank Professor Dr Ilme Schlichting for her many years of service as a Main Editor of JSR, and wish her well as she steps down at this time.While Kristina has been active as a Co-editor of"} +{"text": "This article when published neglected to mention Dr Valerie Moran's second affiliation. She is affiliated with the Luxembourg Institute of Health, Strassen, Luxembourg and the Luxembourg Institute of Socio-Economic Research, Esch-sur-Alzette, Luxembourg."} +{"text": "Dear Readers,The recent science-packed AF Symposium held in Boston, MA was attended by more than 1,200 people from hospitals and health care industries in the United States and around the world and showcased novel atrial fibrillation (AF) ablation techniques and technologies by way of lectures, debates, live case presentations, and posters and abstracts. One of the highlights of the symposium this year was the late-breaking clinical trials. Of particular interest, two out of the four clinical trials presented covered novel energy sources for ablation.Dr. Vivek Reddy of Mount Sinai Hospital in New York, NY presented outcomes of a first-in-human study on the use of an automated noncontact ultrasound imaging and ablation system for the treatment of AF. This novel tool uses low-intensity collimated ultrasound as an energy source. The robotic catheter moves under computer control and allows for both anatomical mapping of the left atrium and ablation. It relies on automated tissue thickness detection and other tissue characteristics to create continuous linear lesions without tissue contact. The catheter system is not yet approved by the United States Food and Drug Administration (FDA); the initial investigation was performed in the Czech Republic and enrolled 52 patients with drug-refractory, symptomatic paroxysmal AF. Manual radiofrequency ablation touch-up was required in a small number of patients. The primary safety endpoint of the study was met, with no occurrence of atrioesophageal fistula, pulmonary vein stenosis, cardiac perforation or tamponade, death, stroke, myocardial infarction, or thromboembolism. There was one instance of phrenic nerve injury. The effectiveness results were impressive at 12 months later, with 80% of the study subjects demonstrating chronic procedure success, defined as freedom from symptomatic atrial arrhythmia recurrence of more than 30 seconds, new class I or III antiarrhythmic drug use, and/or the performance of reablation after a three-month blanking period.Separately, results of the Cryocure II study were presented by Dr. Tom De Potter of OLV Hospital in Aalst, Belgium. Similarly, the multielectrode catheter system used in this research is also not FDA-approved and the study was conducted in Europe. The catheter system uses near-critical nitrogen ultra-low-temperature technology to create deep and transmural lesions in a rapid fashion. Forty-eight patients with persistent and paroxysmal AF were enrolled. In addition to pulmonary vein isolation, patients with persistent AF underwent mitral isthmus ablation and cavotricupid isthmus ablation as needed. Freedom from AF was 100% in those with paroxysmal AF at 12 months and 94% in those with persistent AF at six months, respectively.Importantly, though the research presented in the late-breaking clinical trials session was very impressive, like with any other new technology, the above preliminary results will need to be replicated in larger multicenter studies.The Journal of Innovations in Cardiac Rhythm Management interesting.I hope that you find this issue of Sincerely,md, fhrs, faccMoussa Mansour, Editor in ChiefThe Journal of Innovations in Cardiac Rhythm ManagementMMansour@InnovationsInCRM.comDirector, Atrial Fibrillation ProgramJeremy Ruskin and Dan Starks Endowed Chair in CardiologyMassachusetts General HospitalBoston, MA 02114"} +{"text": "The movement of evidence-based interventions into routine institutional settings like nursing homes is challenging. Among non-pharmacological interventions to address behavioral problems of residents with dementia, Music and Memory (M&M), a popular individualized music listening program, has been shown to have potential to improve quality of life among residents. To examine facilitators and barriers to implementation and sustainability of the M&M program in nursing facilities, a statewide (online and mail) survey of nursing homes was conducted in Wisconsin where the statewide implementation of the program occurred. The response rate was 41% (N=161). Descriptive statistics and content analysis were conducted. Over 80% of facilities provided the M&M program, and 86% of them planned to continue the program. The majority of respondents found the M&M to be beneficial to residents but also reported that the program was not equally effective for everyone, and M&M was time and labor intensive. Barriers to sustainability were: lack of buy-in by direct care staff, use of technology, costs of equipment, inconsistent volunteers, and families not supportive or helpful. Facilitators were: support of facility personnel, family, and volunteers; observing positive effects of program, M&M training provision and support, family involvement, and accessibility of equipment. Targeted resident selection is needed to identify the residents most likely to benefit from the program to avoid possibility of increased agitation or discomfort. Careful consideration is needed for facilities to identify realistic costs, labor, and staff buy-in to promote success."} +{"text": "Research on electronic cigarettes is an emerging field, with the number of articles in this field noted to have grown exponentially over recent years. We used a bibliometric analysis method to analyze the emerging trends and research hotspots in this field.Publication data on electronic cigarettes from 2010 to 2018 were retrieved and downloaded from the PubMed database. Theme trends and knowledge structures were analyzed on the relevant research fields of electronic cigarettes by using a biclustering analysis, strategic diagram analysis, and social network analysis methods. Research hotspots were extracted and compared from three periods.Core topics that have continuously develop between the years 2010 and 2018 include: tobacco use cessation devices; tobacco products; tobacco use cessation devices/adverse effects; smoking prevention and adverse effects; electronic nicotine delivery systems/economics; and public health. Some currently undeveloped topics that could be considered as new future research directions include: tobacco use disorder/therapy; tobacco use disorder/epidemiology; students/psychology; students/statistics and numerical data; adolescent behavior/psychology; nicotine/toxicity; nicotinic agonists/administration and dosage; and electronic nicotine delivery systems/legislation and jurisprudence.Results suggest that some currently immature topics in strategic coordinates and emerging hotspots in social network graphs can be used as future research directions. There is growing evidence that, even though e-cigarettes may produce fewer toxic substances than traditional cigarettes, they may still pose health risks to smokers and people around them3. The long-term effects of e-cigarettes on health are not currently apparent. Moreover, there is insufficient evidence to show that such products may assist people to quit traditional cigarettes4.An electronic cigarette (e-cigarette) refers to a cigarette consisting of a battery, an evaporation heating device, and a tobacco tube containing a liquid smoking product. The nicotine-containing tobacco liquid can be turned into vapor by nebulization for the user to inhale6. However, e-cigarettes may contribute to adverse reactions in the respiratory system8, cardiovascular system9, liver10, and nervous system11. Ever since e-cigarettes became popular among adolescents, the number of studies on e-cigarettes has increased13. In recent years, researchers have become increasingly concerned about tobacco-use disorders14. Some studies have shown that comprehensive interventions are needed to help protect adolescents\u2019 mental health16. With the development of bibliometrics comes its widespread use in health topics. However, there is only a limited amount of bibliometric analyses that focus on tobacco, and we have found only two that focused on e-cigarettes18. This study used a co-word analysis instead of the co-citation analysis applied by the above two articles, to examine trends in e-cigarette research.A large number of recent studies have shown that, from a toxicological standpoint, e-cigarettes as a substitute for nicotine, compared to traditional smoking, may aid in improving public healthMedical Subject Headings) terms/subheadings from which were calculated the number of high-frequency major MeSH terms/subheadings, using the Donohue equation19:This study used the Bibliographic Item Co-Occurrence Matrix Builder (BICOMB) to extract data for the three periods 2010\u20132012, 2013\u20132015 and 2016\u20132018, from PubMed including journals, countries, authors, and major MeSH to construct a social network analysis (SNA) in order to further analyze the knowledge structure of the e-cigarette field. Applying NetDraw 2.084, the major MeSH terms/subheadings network was able to be presented in a 2D map. The nodes represent the major MeSH terms/subheadings, and the links represent their co-occurrence frequencies. Each significant MeSH term/subheading can be evaluated by three parameters, namely, degree, closeness, and betweenness. Degree refers to the number of other nodes directly connected to a node. The higher the degree centrality of a node, the more critical it is to indicate its position in the network. Betweenness indicates the number of shortest paths through a node. The more times a node acts as an intermediary node, the higher is its significance in evaluating the importance of a node in the network. Closeness reflects the closeness between a node and other nodes in the network. It is the sum of the reciprocal of the shortest distances from a specific node to all other nodes in the network. This means that the higher the closeness, the shorter is the distance from this node to other nodes in the network. It is worth noting that the betweenness index was chosen as an evaluation index for the in-depth study of the e-cigarette field.We retrieved and analyzed 54, 968, and 2406 publications for each period, respectively. The number of articles related to e-cigarettes has increased from 8 in 2010 to 988 in 2018, nearly 120 times during the past nine years. Tobacco Control; Nicotine & Tobacco Research; and Przeglad Lekarski; which accounted for 27.3% of the total publications. In the second time period (2013\u20132015), Nicotine & Tobacco Research ranked first, and Przeglad Lekarski was replaced by BMJ . In the third time period (2016-2018), the top three journals were: Tobacco Control; Nicotine & Tobacco Research; and Addictive Behaviors. Overall, Nicotine & Tobacco Research has published the most articles in the field of e-cigarettes over the last nine years.In the first time period (2010\u20132012), the top three journals that published articles on e-cigarettes were: According to the publications retrieved for the three time periods, there were respectively 15, 26 and 49 high-frequency major MeSH terms/subheadings with a total frequency of occurrence of 49.6%, 49.5% and 50.4%, respectively. We considered these as the research hotspots for the three time periods. A biclustering analysis leads to a division of MeSH terms into 4, 3 and 5 clusters for each period, respectively . Biclust22 explains the meaning of strategic diagrams , playing the most important role in the network. In addition, the closeness value is 13, indicating that it is the most closely connected to other nodes. In addition, Tobacco Use Cessation Devices/adverse effects, Smoking Cessation/methods, and Smoking Prevention, also have high degrees of betweenness, indicating that they also have key intermediary roles in the network. The average betweenness value of these three topics is 4.333.Compared to the first three years, Electronic Nicotine Delivery Systems/statistics & numerical data has the highest level of betweenness in the second period SNA diagram and seven new major MeSH terms/subheadings emerged, including: Smoking/Psychology; Electronic Nicotine Delivery Systems/psychology; Smoking/adverse effects; Electronic Nicotine Delivery Systems/adverse effects; Smoking/epidemiology; Electronic Nicotine Delivery Systems; and Nicotine/administration & dosage. At the same time, nine new nodes emerged at the edge of the network, including: Nicotine/analysis; Nicotine/adverse effects; Public Health; Electronic Nicotine Delivery Systems/economics; Smoking Cessation/psychology; Smoking/legislation & jurisprudence; Adolescent Behavior; Tobacco Products; and Tobacco Products/statistics & numerical data. These are considered as emerging hotspots in the field of e-cigarettes in 2013\u20132015.As demonstrated in the 2014\u20132018 SNA diagram, there are six new major MeSH terms/subheadings, including: Vaping/adverse effects; Electronic Nicotine Delivery Systems/methods; Tobacco Products/statistics & numerical data; Smoking Cessation/statistics & numerical data; Smoking Cessation/psychology; and Health Knowledge Attitudes and Practice. There are eight new nodes at the edge of the network, namely: Tobacco Use Disorder/therapy; Tobacco Use Disorder/epidemiology; Students/psychology; Students/statistics & numerical data; Adolescent Behavior/psychology; Nicotine/toxicity; Nicotinic Agonists/administration & dosage; and Electronic Nicotine Delivery Systems/legislation & jurisprudence. These were emerging hotspots in the field of e-cigarettes in the period 2014\u20132018.As our knowledge of e-cigarettes continues to increase, the amount of related research also continues to grow and e-cigarette research has become an emerging field. Using a biclustering analysis, strategic diagrams, and social network analysis diagrams, we analyzed, in detail, the evolution of thematic trends and knowledge structures in the field of e-cigarettes over the past nine years. This is the first time co-word analysis was used to analyze trends in this field.Nicotine & Tobacco Research, and Tobacco Control, are the journals that have published the most articles, however the current study only analyzed the number of e-cigarette articles published in a journal, which is greatly affected by the total number of articles published in a journal. We note that although some journals have a smaller total volume of articles, they also publish in the field of e-cigarettes and have considerable influence, such as Tobacco Induced Diseases, and Tobacco Prevention and Cessation etc.The current study examines e-cigarette publications, globally, by comparing three time periods in the past nine years (2010-2018). Over that period of time, the number of publications related to e-cigarettes has grown rapidly, with England and United States leading the way with the most published articles. 23. Clusters 1, 2 and 3 are in quadrant III. Research in these clusters focuses on: Smoking Prevention and adverse effects; Tobacco Use Cessation Devices/adverse effects; Tobacco Products; Tobacco Use Cessation Devices; and Smoking Cessation/legislation & jurisprudence. These topics are immature and are located in the periphery of e-cigarette research during this period. They may gradually shift to a central and/or mature position, if further research is undertaken. Studies have focused on adverse effects of smoking, and some have now been widely recognized, e.g. smoking cigarettes is the strongest risk factor for chronic obstructive pulmonary disease (COPD)24 and remains the primary risk factor also for lung cancer25. Legislators are increasingly recognizing the dangers of tobacco products as well as the importance of tobacco control and protecting the public from the harm of tobacco26. With the development of e-cigarettes, there is increased concern about their safety as cessation devices, for which there is significant debate; however others have noted that they may provide a potential to quit smoking27.Strategic diagrams were used to analyze the theme trends of publications of three time periods. In the first time period (2010\u20132012), only cluster 0 is in quadrant I, which includes Nicotine/administration & dosage, and Smoking Cessation/methods. We consider these two themes to be research hotspots in the field of e-cigarettes during this period. The research in this period suggests that there is a need for more effective drugs to help smokers quit smoking. During this period, some important research focused on smoking cessation methods and compared various methods including nicotine replacement therapy (NRT), bupropion and varenicline, and nortriptyline and clonidine28, or to quantify how smokers evaluate the attributes of e-cigarettes29. In addition to paying attention to the harm of Tobacco Products, it was also found that e-cigarettes also have the potential to do harm because they contain nicotine, which is addictive and can cause adverse reactions30. Therefore, both tobacco products and e-cigarettes should be treated with caution31. The topics of Tobacco Use Cessation Devices/adverse effects, and Smoking Prevention and adverse effects, gradually developed into the center of the field of e-cigarettes while the adverse reactions of e-cigarettes attracted more attention. Additionally, there were new and immature topics during this period, such as Electronic Nicotine Delivery Systems/economics, and Public Health. Due to increasing attention, these two new keywords have been continuously developed.In the second time period (2013\u20132015), Tobacco Use Cessation Devices, and Tobacco Products, were noted as two themes gradually maturing. In the last period (2016\u20132018), researchers had high hopes for the role of Tobacco Use Cessation Devices in smoking cessation. In this period, research was more extensive, for example, with the aim to apply large cross-sectional surveys to assess the effectiveness of e-cigarettes as an aid to smoking cessationin vivo and in vitro that have demonstrated the harm of e-cigarettes34. The three previously immature topics of Electronic Nicotine Delivery Systems/economics, Public Health, and Tobacco Products, evolved into mature topics. The journal Tobacco Control published several articles about Electronic Nicotine Delivery Systems/economics, and Public Health, in succession37. The journal Tobacco Prevention and Cessation had a special supplement on vape shops39. Clusters 1, 2 and 4, including: Students/psychology; Electronic Nicotine Delivery Systems/psychology; Electronic Nicotine Delivery Systems/legislation & jurisprudence; Tobacco Use Disorder/therapy; Students/statistics & numerical data; Cigarette Smoking/epidemiology; and Electronic Nicotine Delivery Systems/statistics & numerical data, are immature and need further research. During this period, there is an increasing number of studies on the epidemiology of tobacco and statistical analysis of e-cigarettes40. People are paying more attention to the connection between smoking and psychology, particularly the impact of e-cigarettes on youth psychology. In addition, from a social perspective, people also attach great importance to the legislation and jurisprudence governing e-cigarettes41.The strategic diagram for the third time period (2016\u20132018) describes the knowledge structure of the e-cigarette field and provides a large amount of information on emerging, prominent research. Custer 0, Electronic Nicotine Delivery Systems/adverse effects have been developed at this stage. With the popularity of e-cigarettes growing, there was an increase in experiments Three SNA diagrams were made according to the high-frequency MeSH terms/subheadings. In these three time periods, 4, 10 and 16 major MeSH terms/subheadings, respectively, had a high degree of centrality. Nicotine/administration & dosage in the first time period, and Electronic Nicotine Delivery Systems/statistics & numerical data in the second and third periods are at the center of the SNA diagrams and have the greatest number of direct connections to other nodes, suggesting the most significant impact during each period.In addition, in the second period, there are nine MeSH terms at the edge of the network that are new and immature. Among these, Nicotine/analysis, Nicotine/adverse effects, and Public Health became developed in the third period. We can consider these MeSH terms as emerging hotspots in the second period (2013\u20132015). Similarly, Tobacco Use Disorder/therapy, Tobacco Use Disorder/epidemiology, Students/psychology, Students/statistics & numerical data, Adolescent Behavior/psychology, Nicotine/toxicity, Nicotinic Agonists/administration & dosage, and Electronic Nicotine Delivery Systems/legislation & jurisprudence can be considered as hotspots in the most recent of the analyzed years (2016-2018).To the best of our knowledge, the current study is the first to use a co-word analysis method to perform a comprehensive analysis of e-cigarette publications. The e-cigarette field is constantly evolving, and there will be more in-depth research in the future. It is our contention that the emerging hot issues mentioned above can guide clinicians and researchers to develop new projects in the e-cigarette area. At the same time, this research has certain limitations. The first is that we only searched for journals, excluding comments and other types of literature, and perhaps missed some research hotspots. Secondly, co-word analyzes high-frequency MeSH terms only, which may affect the results of the cluster analysis as our results are based on the number of articles published in each medium and not the impact of each article. Moreover, in the future, we could use a variety of databases for analysis, such as Cochrane, Embase, clinical trials.gov, some guidelines could also be searched, as well as manual searching for grey literature.We applied a biclustering analysis, strategic diagram and SNA methods, to analyze high-frequency MeSH terms, and conducted a co-word analysis on the field of e-cigarettes. This research shows that Tobacco Use Cessation Devices, Tobacco Products, Tobacco Use Cessation Devices/adverse effects, Smoking Prevention and adverse effects, Electronic Nicotine Delivery Systems/economics, Public Health, and Tobacco Products, are the core topics that constantly evolved between the years 2010 and 2018. Tobacco Use Disorder/therapy, Tobacco Use Disorder/epidemiology, Students/psychology, Students/statistics & numerical data, Adolescent Behavior/psychology, Nicotine/toxicity, Nicotinic Agonists/administration & dosage, and Electronic Nicotine Delivery Systems/legislation & jurisprudence can be considered as hot research topics in the analyzed period 2016\u20132018.Click here for additional data file.Click here for additional data file."} +{"text": "Nature Communications 10.1038/s41467-020-14500-z, published online 7 February 2020.Correction to: The original version of this Article contained an error in the Competing interests section, which incorrectly omitted \u201cT.I. is a co-founder of Data4Cure and has an equity interest. T.I. is on the Scientific Advisory Board of Ideaya BioSciences, Inc., has an equity interest, and receives income. The terms of these arrangements have been reviewed and approved by the University of California San Diego in accordance with its conflict of interest policies\u201d. This has been corrected in both the PDF and HTML versions of the Article."} +{"text": "Tuberculosis (TB) depicts heterogeneous spatial patterns with geographical aggregation of TB cases due to either ongoing person-to-person transmission or reactivation of latent infection in a community sharing risk factor. In this regard, we aimed to assess the spatiotemporal aggregation of drug-resistant TB (DR-TB) patients notified to the national TB program (NTP) from 2015 to 2018 in selected districts of Karnataka, South India.2 with more than the expected count of DR-TB patients were constructed.This was a cross-sectional study among DR-TB patients notified from Dakshina Kannada, Udupi, and Chikamagalur districts of the state of Karnataka. Clinico-demographic details were extracted from treatment cards. The registered addresses of the patients were geocoded (latitude and longitude) using Google Earth. Using the QGIS software, spot map, heat maps and grid maps 25 kmOf the total 507 patients studied, 376 (74%) were males and the mean (standard deviation) age of the study participants was 41.4 (13.9) years. From 2015 to 2018, the number of patients increased from 85 to 209 per year, the area of aggregation in square kilometers increased from 113.6 to 205.7, and the number of rectangular grids with more than the expected DR-TB patients (> 1) increased from 12 to 47.The increase in the number of DR-TB patients, area of aggregation, and grids with more than the expected count is a cause for concern. The NTP can use routine programmatic data to develop maps to identify areas of aggregation of disease for targeted TB control activities. Globally, tuberculosis (TB) remains a major public health problem of concern with an estimated 10 million incident TB patients and 1.3 million deaths due to TB in the year 2017. Multidrug-resistant tuberculosis and extensively drug-resistant tuberculosis have emerged globally and pose a threat to TB control efforts. The World Health Organization (WHO) estimated about 330,000 incident multidrug-resistant TB or rifampicin-resistant TB (MDR/RR-TB) patients globally in the year 2017 [India is one of the 30 high-burden TB countries and has a triple burden of TB, TB/HIV, and MDR-TB. In 2017, it was estimated that there were 65,000 MDR/RR-TB patients in the country with a prevalence of 2.8% among new TB cases and 12% among previously treated TB cases. Similar to global trends, there is a substantial gap in the detection and treatment of MDR/RR-TB patients in India. Of the estimated, only about 40% of the MDR/RR-TB patients are identified and initiated on treatment [TB depicts heterogeneous spatial patterns with localized aggregation of cases due to either ongoing person-to-person transmission or reactThough geospatial analysis, identifying areas with the aggregation of TB patients, and focused TB control activities in such areas may be theoretically beneficial, such explorations are scarce. As a first step, there is a need for assessing the geospatial distribution of drug-resistant TB locally to detect whether there is any geographical aggregation of DR-TB patients using routine program data. However, the inherent difficulties in geotagging the location of the patient and technical challenges in handling such data have limited the use of geospatial epidemiology in TB control efforts. Hence, as a desk review, we explored the possibility of geocoding the address of DR-TB patients using Google Maps and constructing the spatial heterogeneity maps. We geocoded all DR-TB patients treated under the Revised National Tuberculosis Control Program (RNTCP) in three selected districts of Karnataka from 2015 to 2018 to depict the spatiotemporal pattern of these patients.It is a cross-sectional descriptive study using secondary data collected routinely by the RNTCP of India.Karnataka is a southern state of India. The approximate population of Karnataka is 66.8 million with a sex ratio of 973 females to 1000 males [The study was conducted in the DR-TB center of Mangalore, Karnataka. The DR-TB center caters to all the DR-TB patients diagnosed in three districts, namely Dakshina Kannada, Chikmagalur, and Udupi Fig. . AccordiThe Programmatic Management of Drug-Resistant TB (PMDT) guideline was followed for the diagnosis and treatment of DR-TB in the study site. From 2015 to 2017, the criteria-C was adopted for identifying presumptive MDR-TB patients . All theAll the diagnosed DR-TB patients from the districts included in the study were referred to the DR-TB center at Mangalore for pretreatment evaluation and initiation of treatment. The patients reaching the DR-TB center were registered for care with a unique DR-TB registration number. The socio-demographic and clinical details, along with the address and phone number of the patient, were recorded in the DR-TB register by the staff nurse. Also, the PMDT treatment card was issued for each patient and had the same details recorded in it.At the DR-TB center, the patients were hospitalized for pretreatment evaluation and treatment initiation. All DR-TB patients were prescribed drugs based on PMDT guidelines. After initiating the therapy, patients were monitored at the DR-TB center for 1 to 2 weeks. Once a patient is stable and tolerating the second-line drugs, then he/she was referred to the peripheral health institute (PHI) nearest to patient residence for further care and monitoring with ambulatory DOTS-Plus treatment at PHI. All the details of the treatment course in the DR-TB center and referral details were updated in the DR-TB treatment card and DR-TB register.The DR-TB patients diagnosed by private providers are notified to RNTCP. As the treatment of DR-TB is complicated, majority of the patients are referred to public health facilities for treatment and eventually initiated treatment at the DR-TB center. However, a few patients might continue to be treated by private providers.All DR-TB patients initiated on TB treatment from a DR-TB center at Mangalore from January 2015 to December 2018 were included.Data on socio-demographic and clinical characteristics and drug-resistant pattern like PMDT number, age, gender , village/ward, taluk, district, name of TU, name of PHI, DOTS provider, date of DST, date of DST results, date of registration, type of TB (new/treatment after loss to follow-up/treatment after failure/recurrent/relapse), site of TB (pulmonary/extra-pulmonary), type of DST used for diagnosis (CDST/Xpert MTB/RIF assay), molecular test results (not done/Mtb+/Rif+/Mtb+/Rif\u2212/Mtb\u2212/MTB+ Rif indeterminate), type of DR-TB (RR-TB/MDR-TB/XDR-TB), resistance to isoniazid (yes/no/not available), resistance to ethambutol (yes/no/not available), resistance to pyrazinamide (yes/no/not available), resistance to streptomycin (yes/no/not available), resistance to fluoroquinolones (yes/no/not available), resistance to kanamycin (yes/no/not available), HIV status (yes/no/not available), ART status (yes/no/not available/not applicable), weight at initiation of treatment, tobacco use (yes/no/not available), and alcohol use (yes/no/not available) were extracted from DR-TB treatment card and DR-TB register maintained at DR-TB center.The geocode (latitude and longitude) of each patient was obtained based on the registered address using Google Earth pro version 7.3.2.5776 application. The village/ward/street of the patient was used to geocode the address \u201317. The Data were double entered and validated using EpiData Entry software and analyzed using EpiData analysis and Stata version 12.0 . Socio-demographic and clinical characteristics and drug-resistant patterns were summarized using numbers and percentages.The QGIS version 2.18.15 ) was used to plot the drug-resistant TB patients, and a map of geospatial distribution was constructed. The heat maps depicting spatial heterogeneity were con16) was u2. Within each grid, the number of DR-TB cases was counted. Those cases with more than the expected number of DR-TB cases were colored red. The expected DR-TB patients per grid were calculated with a population density of 282 per square kilometer in three districts, and the estimated number of TB patients per 25 km2 is 15 [2 assuming the proportion of DR-TB cases among TB patients to be 6%.The grid maps were constructed with each grid of 25 kmcountry) . The expIn total, 507 DR-TB patients were notified and initiated on treatment from 2015 to 2018 in the study districts. The mean (SD) age of the study participants was 41.4 (13.9), and 376 (74%) were males. Of the total, about 452 (89%) of the DR-TB patients were new TB patients. The DR-TB patient characteristics included in the study are shown in Table 2 with more than the expected DR-TB patients (> 1) increased from 12 to 47 during the year 2015 to 2018 , there was an increase in the number of DR-TB patients, the sum of the area of aggregation of patients, and the number of grid units with more than the expected DR-TB patients in the study districts. About one in ten DR-TB patients had resistance to fluoroquinolone.There was an increase in the number of DR-TB patients in the study districts. The increasing trend was noted in each of the study districts. The annual TB reports have also reported an increase in notified DR-TB patients from 2015 to 2017. The increase might be either due to improved care delivery and adherence to protocols for the detection of DR-TB or due to the uninterrupted transmission of DR-TB. The massive increase in the number of DR-TB cases in 2018 could be due to introduction of the universal DST . Improving the diagnostic services and making the services accessible might have increased the number of notified DR-TB cases. The other potential reasons for the increase in notification of DR-TB patients are imporved quality of DR-TB in public sector and also the disinterest of private providers in treating DR-TB patients within the private sector.The previous studies in Peru and MoldThe study has a few strengths. First, geocoding was done for all the notified cases during the study reference period as all the addresses could be extracted and traced back. Hence, there was no selection bias due to missing information in routine programmatic records. Second, the individual DR-TB patients were geocoded. This limited the aggregation bias of constructing heat maps using administrative boundaries and provided the opportunity to construct the heat maps based on the occurrence of an event. Third, the sample size was good for conducting spatial heterogeneity analysis and also spatiotemporal analysis. Fourth, double data entry and validation were used. This helped to limit the data entry errors and improved quality of data, more so with variables like latitude and longitude with five digits after the decimal point.2 was calculated based on the estimated TB incidence rate for India and averaging the population density in the three districts. This might have reduced the internal validity of the study findings. However, the incidence rate in the study districts might be lower than that of the country estimates, and thus, the expected number of DR-TB patients is an overestimate. The population across the district is not uniform, and there might be areas with a higher population density with a relatively higher number of expected DR-TB patients leading to underestimation. Thus, on the whole, the estimation of expected DR-TB patients per 25 km2 might be close to a real-world scenario. Third, there were no village- and ward-level shapefiles, which might have helped us to calculate the rates of occurrence of the DR-TB. Hence, we had to calculate the expected number of DR-TB patients, which might be inferior to rates. Fourth, the data series was available only for 4 years and had extreme variation. This restricted the use of time series analysis and prediction models. Fifth, we might have missed the undetected and not notified DR-TB patients. This could have limited the validity of heat maps as the not notified and undetected case might contribute to disease transmission but not accounted for in the analysis. Sixth, the study might have failed to capture the natural trend in the occurrence of the DR-TB as there was a change in the diagnostic algorithm, which could have influenced the case detection in the year 2018.The study has a few limitations. First, the geocodes were approximated to village/ward or street level and thus failed to geotag the house of the patient. However, the villages in this part of the country have a small area and therDespite several limitations, the study has a few implications. First, over the years, there was an increase in the number of cases and also in the area of aggregation of cases. The RNTCP needs to explore the potential reasons for this increase in DR-TB patients. If the programmatic change of upfront DST has brought this change, then this is a promising step in efforts towards control of DR-TB in India. However, if the increase is due to the transmission of the disease, then this is a cause for concern. Second, the geocoding and generation of heat maps provided insights on the aggregation of disease and the spread of disease to new areas over the years. The program can train the data entry operators to geocode the villages of DR-TB patients and generate maps to help in decision-making. Third, the utility of concentrated TB control efforts in areas with DR-TB aggregation on reducing the transmission and burden of DR-TB needs to be explored. Fourth, on moving towards the elimination of TB, there is a need for developing the geospatial monitoring indicators like \u201cnumber of TB or DR-TB case per square kilometer\u201d area and prioritize areas with high number of cases for intensified TB control activities.The increase in the number of DR-TB patients, area of aggregation, and grids with more than the expected count is a cause for concern. The NTP can use routine programmatic data to develop maps to generate local foci for targeted TB control activities."} +{"text": "Aim: Although criteria and recommendations for the successful integration of sex- and gender-sensitive aspects in medical teaching have already been published, only a few medical faculties in Germany have conducted the systematic integration of sex- and gender-sensitive medicine. The aim of this expert survey, therefore, was to describe the current approaches to the integration of sex- and gender-sensitive medicine in teaching in the sense of Good Practice.Method: Between April and June 2018, guided interviews were conducted with nine experts in the field of sex- and gender-sensitive medicine. Each of the experts had had experience of implementing sex- and gender-sensitive medicine at their universities. The expert interviews were then evaluated by means of quality content analysis, and frequency analyses were carried out.Results: Aspects of sex- and gender-sensitive medicine were integrated both longitudinally and selectively into the compulsory curriculum or elective fields of various medical, health and nursing science courses. In the opinion of the experts, medical studies should promote the students\u2019 gender sensitivity and in particular impart knowledge about the psychosocial and biological aspects of sex- and gender-related differences and sex- and gender-sensitive communication. For the methodological implementation of the integrated contents, didactic resources were partly adapted or developed. The players in the implementation process were confronted with various challenges, e.g. the involvement of the lecturers, the perception of sex- and gender-sensitive medicine as a women\u2019s theme as well as ensuring the sustainable integration of sex- and gender-sensitive medicine, which is also structurally anchored in the faculty. Aspects of the curricular integration and the structural anchoring were mentioned i.a. as being crucial for success. A combination of top-down and bottom-up processes, e.g. by involving the faculty management but also by supporting student initiatives, was described as conducive to success. Conclusion: The depicted approaches to the integration of sex- and gender-sensitive teaching contents give insight as to how sex- and gender-sensitive medicine can be integrated into the curricula. The interviews with the experts point to current themes related to sex- and gender-sensitive medicine and didactic resources. Moreover, it becomes clear which challenges are to be expected for the integration of sex- and gender-sensitive medicine in teaching and how these can be addressed. Particularly the involvement of the faculty\u2019s lecturers but also the sustainable integration and continual quality assurance of sex- and gender-sensitive contents present challenges of a crucial nature. Based on the biopsychosocial model, sex- and gender-sensitive medicine concerns itself with the influence of the biological sex and psychosocial/sociocultural gender with regard to the emergence, diagnosis, therapy and prevention of illnesses and has the overriding goal of ensuring the best possible healthcare for all gender , 2]. Th. Th2]. TA necessary prerequisite for healthcare that is in keeping with the times is taking into account the variables \u201csex\u201d and \u201cgender\u201d as important determinants of health and disease . In ordehttp://www.gender-curricula.com/gender-curricula-startseite/]) were contacted. In addition, experts were also recruited via calls for studies and direct contact with networks dealing with sex- and gender-sensitive medicine and gender research. Experts in the field of sex- and gender-sensitive medicine were informed about the study via e-mail. The e-mail invitation for the experts\u2019 interviews contained information about the study objectives as well as on the themes and procedure of the interviews so that the contacted experts were also in a position to cross-check the required expertise. A total of 18 experts were invited to take part in the interviews. In the present study, interviews were conducted with experts who had already been involved in processes implementing sex- and gender-sensitive medicine into medical, healthcare and nursing science teaching. The experts were recruited between February and May 2018. Based on literature and internet research, contacts for teaching projects in the field of sex- and gender-sensitive medicine at medical faculties were approached and experts working in the fields of sex- and gender-sensitive medicine and nursing science , B.A. in Nursing and Health Promotion (n=1), M.A. in Nursing Science (n=1), Dentistry (n=3), Molecular Medicine (n=2), Public Health (n=2), M.A. in Health Professions Education (n=1), M.A. in International Health (n=1), PhD programme in Medicine/Dentistry (n=1), and in a habilitation course (n=1). In three universities a longitudinal integration has been achieved, for example in all the module manuals, in specific events concerning sex- and gender-sensitive medicine and in compulsory lectures of a \u201cBasic Curriculum Gender Medicine\u201d. In four other universities, sessions on sex- and gender-sensitive medicine have been selectively anchored, for example as a seminar in compulsory courses , as an elective subject, in individual modules or in a series of interdisciplinary lectures. Summarizing, four universities had a combined integration in the compulsory courses and in elective subjects or modules. Two universities had integration in the compulsory courses and one university in the elective courses. At one university an open optional workshop series had been conducted. rd/10th semester), in a multiple choice exam at the end of the study block, as a written elective exam or a conference article (PhD). One of the universities is planning to query the sex- and gender-sensitive content in the OSCE. Moreover, one of the universities is offering the opportunity of acquiring a key qualification by completing a basic curriculum in Gender Medicine (22 compulsory lectures with integrated sex- and gender-sensitive content).The integrated aspects of sex- and gender-sensitive medicine are tested in various ways, e.g. in the semester exams or through the evaluation of teaching practices. In addition, the learning objectives formulated during the introduction of the model study programme were enumerated. Surveys with the students were also mentioned as quality assurance measures as were departmental conferences with module coordinators, the sustainable safeguarding of the learning objectives, higher education didactics for lecturers, and the anchoring of the implemented contents in the directives of the system accreditation.The The integrated sex- and gender-sensitive contents were often developed from the experts\u2019 own field of expertise or from their research, were the result of benchmarking processes at other universities, or were based on surveys within the faculty or expert panels, for example with specialists or equal opportunity officers. Sex- and gender-sensitive themes were also selected according to their topicality, evidence-basing and the possibility for reflection and discussion (e.g. gender paradox). Available catalogues of learning objectives, e.g. from the Association of Professors of Gynecology and Obstetrics (AGPO) were adahttps://gendermedwiki.uni-muenster.de/mediawiki/index.php/Willkommen_bei_GenderMed-Wiki] were applied. Various teaching methods e.g., case studies, problem-oriented learning (POL), film material, discussions , communication training, developing counseling concepts taking the variables \u201csex\u201d and \u201cgender\u201d into account, interviews (with patients/experts), hands-on training, blended learning or research assignments were used in the lectures, seminars and working in small groups that were carried out in the study courses. Didactic resources such as the German version of the \u201cGender Lens Tool\u201d or the GWith regard to the further development of sex- and gender-sensitive medicine in teaching, the experts would like the subject to be integrated systematically as an interdisciplinary topic in medical training right from the beginning of the study courses and for it to be reflected in the examinations. The individual medical departments should provide the necessary information, a specific elective would then be unnecessary. Furthermore, sex- and gender-sensitive medicine should be increasingly considered in research in order to guarantee broader empirical data. Research findings should then be transferred into teaching, and current literature and guidelines should be critically reflected.In addition, when it comes to gender issues, medicine should become more closely networked with other disciplines, e.g. gender studies, and sex- and gender-sensitive medicine should become institutionally anchored, e.g. through a coordination office for sex- and gender-sensitive medicine. Sex- and gender-sensitive medicine has already been integrated in various study courses for medicine, healthcare and nursing science, which confirms the relevance of the subject for the healthcare professions. Many of the experts have stated that sex- and gender-sensitive medicine should be understood as an interdisciplinary cross-sectional topic; however, the implementation strategies and the inclusion in the curricula have varied between the universities. Three universities achieved a longitudinal integration of sex- and gender-sensitive medicine. Here the longitudinal implementation was partly planned from the start whereas in other faculties specific teaching units on sex- and gender-sensitive medicine were selected and integrated, some of which will be replaced in the future by continuous integration in the departments or will continue to exist side-by-side. With regard to the integration of sex- and gender-sensitive medicine in teaching, the results of the interviews with the experts confirm the knowledge gained from other international studies. For the longitudinal integration of sex- and gender-sensitive medicine the further development of the existing curriculum is recommended, e.g. the teaching material should be checked for gender-sensitivity, gaps in the curriculum detected and integrated aspects underlined , 5], , , 17], , . In an oA fundamental question arises in the integration of sex- and gender-sensitive medicine into compulsory teaching or elective courses. The advantage of the elective range lies first of all in more flexible and feasible integration. On the other hand, integration would only be selective and hence reach only some of the students .Incorporating the teaching staff in the integration of sex- and gender-sensitive medicine was considered by the experts as being one of the main challenges for the implementation. A lack of sex- and gender-related knowledge in the faculty, limited time resources and interest by the lecturers as well as lacking transparency about what will actually be taught have been described as obstacles in earlier publications , 8], 1, 18], , . Authors, To emphasize the focus of sex- and gender-sensitive medicine on the healthcare of all genders, this should be differentiated concerning the content, e.g. research on women\u2019s health alone and, according to the structure, also be anchored accordingly in the faculty, e.g. not necessarily incorporated in the equal opportunities office , 10]. A. A10]. Ahttps://gendermedwiki.uni-muenster.de/mediawiki/index.php/Willkommen_bei_GenderMed-Wiki].Substantively, the students should, according to the experts, be sensitized first of all towards the themes of sex- and gender-sensitive medicine, particularly the psychosocial aspects of sex- and gender-related differences. For this, core competences should be suggested and registered, and should also be relevant for exams, for instance by being anchored in the German National Competence-Based Learning Objectives for Undergraduate Medical Education (NKLM) , 10], , . For theThe results of the interviews with the experts describe implementation approaches from various universities. The experts state that sex- and gender-sensitive medicine is a relevant cross-sectional theme in many fields and should be integrated sustainably in the curricula. In addition to a fundamental sensitization for aspects of sex- and gender-sensitive medicine , knowledge about sex- and gender-specific differences and practical skills (e.g. sex- and gender-sensitive communication) should be imparted. From the structures and success-critical factors shown in the implementation process it is possible to deduce how the successful integration of sex- and gender-sensitive medicine into teaching , 10], , can be aAt the same time the interviews with the experts showed that the integration of sex- and gender-sensitive medicine into teaching is linked with numerous challenges. The main challenge was described as being the involvement of the lecturers, who were approached in several ways, such as by providing sex- and gender-sensitive teaching material, introducing the subject in faculty meetings or by holding advanced training courses. Problems occurred not only in the initial integration of sex- and gender-sensitive contents but also in the sustainable quality assurance of those contents, which were partly due to a shortage of human resources. The fundamental commitment of the faculty management in relation to the integration of sex- and gender-sensitive contents in the curriculum is therefore essential for success. We would like to thank the experts who participated in the interviews for their support and expertise: PD Dr. med. Anja B\u00f6ckers (Ulm University); Miriam Engels (Heinrich Heine University D\u00fcsseldorf), M.Sc.; Prof. Dr. phil. Margret Flieder (Evangelische Hochschule Darmstadt - University of Applied Sciences); Univ.-Prof. Dr. med. univ. Margarethe Hochleitner ; Dr. rer. medic. Sabine Ludwig (Charit\u00e9 \u2013 Universit\u00e4tsmedizin Berlin); Dr. phil. B\u00e4rbel Miemietz ; Prof. Dr. rer. nat. Dr. med. Bettina Pfleiderer ; Prof. Dr. med. Marianne Schrader (University of L\u00fcbeck) and Dr. phil. Simone Weyers (Heinrich Heine University D\u00fcsseldorf). Our thanks also go to the KoordinierungsstelleGenderforschung & Chancengleichheit Sachsen-Anhalt and to the anna fischer project for helping us to recruit the experts. The authors furthermore thank Vivienne Krause for the English translation of the manuscript. The authors declare that they have no competing interests."} +{"text": "Scientific Reports 10.1038/s41598-019-43680-y, published online 15 May 2019Correction to: The original version of this Article contained errors in the spelling of the authors Cristina B. Adamo and Alexander Flacker, which were incorrectly given as Cristiane B. Adamo and Alexandre Flacker respectively. These errors have now been corrected in the PDF and HTML versions of the Article, and in the accompanying Supplementary Information file."} +{"text": "The first speaker is Dr. Lori Martin-Plank, an established academic at the University of Arizona, College of Nursing. Dr. Martin-Plank will provide her experiences in advocating for older adults in Pennsylvania and nationally through professional organizations, meeting with coalition partners to promote access to care for vulnerable older adults in rural areas by promoting full practice authority for nurse practitioners, and advocating for full home health authority for nurse practitioners. Dr. Martin-Plank will share how she is active in advocacy and policy at the local, state and federal levels, and how to build a presence and relationship with legislators on The Hill and State Capitol. Dr. Martin-Plank is a family, gerontological, and mental health nurse practitioner, practicing in Pennsylvania, New Jersey, and Arizona."} +{"text": "Dear Editor,Rash, urticaria, and varicelliform presentations have already been associated with SARS-CoV-2.4A 39-year-old female patient presented fever, dry cough, and odynophagia two weeks prior. After a presumptive diagnosis of COVID-19, she was treated with azithromycin, acetylcysteine, vitamin C, and zinc. On the tenth day, she presented anosmia, worsening of the cough, and painless, non-pruritic lesions on the fingers.The patient reported hepatic steatosis and systemic arterial hypertension .The physical examination revealed erythematous macules on the third, fourth, and fifth left fingers . Skin biHistopathological examination revealed a discrete focus of spongiosis in the epidermis, with slightly increased volume of keratinocytes, cytoplasmic vacuolization, and elongated and hyperchromatic nuclei. Apparently, there was pale intranuclear inclusion. In the dermis adjacent to these focal areas, there was a slight interface change with lymphohistiocytic permeation of the basal layer, perivascular inflammatory infiltrate, and vessels with intermingled nuclear debris .Figure 3The SARS-CoV-2 virus was not identified by RT-PCR in the samples collected from the nasopharynx and skin. The anti-SARS-CoV-2 IgM antibody in peripheral blood was reactive and the IgG was non-reactive. Serological tests for dengue fever (DENV), Zika (ZIKV), and chikungunya (CHKV) virus were negative.The patient was treated with topical corticosteroids, three times a day, for five days, with disappearance of the cutaneous lesions.In Italy, the majority of SARS-CoV-2 patients developed pernio-like lesions on the tenth day of illness, similar to the present case.There is still no gold standard technique for the identification of SARS-CoV-2 in the skin. In the autopsy of patients with COVID-19, a swab was introduced directly into the lung tissue. In that study, a sample collected was positive, by RT-PCR, for SARS-CoV-2.In patients with skin rash, from tropical countries, several viruses should be investigated; DENV, ZIKV, and CHIKV are among the main ones. In the present case, there was also the possibility of a drug eruption, since in addition to antihypertensive drugs, the patient also used azithromycin and acetylcysteine before the onset of the skin condition. This hypothesis was ruled out by the histopathological examination. The IgM antibody reagent for SARS-CoV-2, negative serologies for DENV, ZIKV, CHIKV, and histopathological findings suggest that the lesions presented here are associated with COVID-19.Patients with a clinical and/or laboratory picture of COVID-19 with cutaneous manifestations should be clinically and histopathologically evaluated by dermatologists, for the correct diagnosis and therapeutic conduct.This case report was submitted and approved by the Research Ethics Committee of Funda\u00e7\u00e3o Alfredo da Matta de Dermatologia (CAAE: 32573520.7.0000.0002). The patient signed an informed consent.None declared.Luciana Botinelly Mendon\u00e7a Fujimoto: Approval of the final version of the manuscript; design and planning of the study; analysis and interpretation of data; critical review of the manuscript.Silvana de Albuquerque Damasceno Ferreira: Approval of the final version of the manuscript; analysis and interpretation of data; critical review of the manuscript.Fabiane Braga dos Santos: Approval of the final version of the manuscript; design and planning of the study; analysis and interpretation of data; critical review of the manuscript.Carolina Talhari: Approval of the final version of the manuscript; design and planning of the study; analysis and interpretation of data; editing of the manuscript; critical review of the manuscript.None declared."} +{"text": "Scientific Data10.1038/s41597-020-00579-y, published online 16 July 2020Correction to: Following publication of this Data Descriptor, it was found that the number of ticks collected was incorrectly stated in the Background & Summary section of the text. The correct number of ticks collected was 29,440. This error has been corrected in both the HTML and PDF versions of this Data Descriptor.In the original figshare dataset, the sites O-035, O-074 and O-078 erroneously had number of ticks collected set to zero, this has been amended. Additionally, ticks were not collected at revisited sites as originally indicated, and this has also been amended in the related figshare record."} +{"text": "Correction to: Trialshttps://doi.org/10.1186/s13063-019-3347-yAfter publication of our article the auth\u201cFirst, S. Bahrami has been involved in the process of our study from the beginning. Second, S. Bahrami worked very hard to achieve the economic part of the protocol - described in the method section \u2018Cost-efficiency analysis\u2019. For recall, an economic assessment is being developed in order to take into account the cost of both interventions in our study (MFT and SFT), to identify any possible difficulties in implementing both interventions, to evaluate all treatments and supports that caregivers of patients with eating disorders could for \u2026 A cost-efficiency analysis will be conducted on the costs generated by the interventions with a perspective on their impact on patients\u2019 health and families\u2019 quality of life. This methodological choice results from the multi-dimensional consequences of the illness on the patients\u2019 physical and psychiatric health and social integration, and on their families.\u201dWith this addition, the Author\u2019s contribution section is also modified to the below:BC is the principal investigator of the study. NG is the scientific director of the study and co-investigator. JD is a co-investigator. CBa is the leading biostatistician. SB is in charge of the economic evaluation. NG, BC, JD, and LR designed the study in collaboration with CBl, MC, SB and MRM. JD and LR drafted the original version of the study protocol with the assistance of ASM, NG, VBC and BC. BC, IK, ZJ, and AH are therapists in SFT and MFT. All authors read and approved the final manuscript."} +{"text": "Coronavirus disease 2019 (COVID-19) is highly contagious, and thus has become an emerging health crisis worldwide. The optimal strategies to prevent the spread of this disease are inconclusive, and therefore, the adopted measurements to combat COVID-19 varies in different countries. In mid-March and late-August 2020, we performed internet searches to collect relevant information, from sources such as the website of the World Health Organization. The epidemiological data of COVID-19 from several countries were collected and we found that Taiwan had a comparably successful story for combating the pandemic. As of mid-March, Taiwan had high rates of diagnostic testing (688.5 tests per million citizens) with a lower infection rate . As of late-August, there were 488 cases (20 cases per million people). Furthermore, Taiwanese government-guided strategies and hospital data were also reviewed. We summarized some important strategies to combat COVID-19, which include: (1) border control; (2) official media channel and press conferences; (3) name-based rationing system for medical masks; (4) TOCC-based rapid triage, outdoor clinics, and protective sampling devices; and (5) social distancing, delaying the start of new semesters, and religious assembly restriction. In conclusion, Taiwan had lower rates of COVID-19 compared with other countries, and Taiwan government-guided strategies contributed to the control of the disease's spread. In December 2019, novel coronavirus disease (COVID-19), caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), was detected in central China and then spread throughout the country and to the rest of the world rapidly . The numDue to the recent emergence of SARS-CoV-2 in humans, researchers are developing best practices in real-time to combat this new virus. Several strategies were adopted very early to block transmission, including the unprecedented lockdown of Hubei and other provinces, and travel bans within China and many other countries globally , 6, 12. Taiwan is a small and populous island which is geographically very close to China. Social interaction between China and Taiwan is frequent, thus leading to a high risk of virus transmission. Central Epidemic Command Center (CECC) was assembled to combat the COVID-19 pandemic in Taiwan on 20 January 2020. The commander of the CECC was the Taiwanese Ministry of Health and Welfare minister, and the organization's members included experts from various fields. The CECC executed several strategies to reduce disease transmission and its government-guided strategies may have contributed to mitigating the disease's spread. Taiwan's first confirmed case was detected on 21 January 2020, and by mid-March there were 49 cases . As of lOur study was approved by the ethical committee of MacKay Memorial Hospital, Taipei, Taiwan (registry number: 20MMHIS140e). As of mid-March 2020, we prospectively searched the websites of WHO, Taiwan Centers for Disease Control (CDC)\u2014which is a key department under the Ministry of Health and Welfare and is responsible for disease prevention and control\u2014and other websites to extract data regarding patient numbers and diagnostic tests of COVID-19 in some countries based on the epidemic conditions and completeness of publicly available data \u201318. The Taiwan's CECC delivered information to the populace via public broadcasting; this included television, newspaper, and the internet . AdditioAs of 13 March 2020, 49 cases were confirmed as COVID-19 in Taiwan with an overall low incidence rate and low infection rates compared with other countries; in fact, it had one of the lowest incidences in the world. Government-guided strategies contributed to controlling the disease's spread and may be beneficial for reference by other countries' health policy makers and healthcare providers.Timely quarantine and identification of infectious sources are essential to reduce virus transmission , 24. HowPublic education with correct information about virus transmission and disease prevention is crucial during a pandemic. The CECC invited famous internet celebrities and YouTubers to make videos in various languages and to share correct information. For health personnel, teleconferences were held to share knowledge and standard procedures of medical care. All these measurements reduce unnecessary fear and panic.The innovative \u201cname-based rationing system for masks\u201d was believed to have contributed to disease control during the COVID-19 pandemic . The majUnnecessary hospital visits were prohibited. Patients who look for medical aid in Taiwan are always asked for TOCC history. At the hospital entrance, a short version of COVID-19 symptoms was posted to provide a simple, graphic, and clear reminder of the disease . People Our study had some limitations. First, we recognize that there is no single best strategy and that the true impact of each strategy remains unclear. The optimal strategy will differ by geographic region, culture, population density, and healthcare resources and norms. Second, due to the lack of relative quantification of the impact for each one of the components that are mentioned as part of the public health strategy, it's difficult to investigate the independent impact of each measurement.In conclusion, the emerging COVID-19 pandemic is an important health crisis worldwide. The number of infected people increased exponentially in many areas, while Taiwan experienced a relatively controllable situation. These government-guided strategies may contribute to the reduction of disease transmission.All datasets generated for this study are included in the article/C-CC, C-YT, W-MC, C-MC, and C-YL were responsible for conception. C-CC, C-YT, Y-CL, C-YH, P-HL, and Y-LT were involved in study methodology and data collection. W-MC, T-HS, and S-LW supervised study. C-YT, Y-CL, and C-MC performed data analysis. C-YL wrote the first draft. C-CC, C-YT, and W-MC contributed to this work equally. All authors approved of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Nature Communications10.1038/s41467-020-17378-z, published online 20 July 2020.Correction to: The original version of this Article contained an error in the author affiliations.The affiliation of Bo Gao with National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA was inadvertently omitted.This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "Kits were made up of DNA extraction and PCR amplification systems based on species-specific, and universal primers. The reference meat mixtures and commercial samples were extracted by the kit and PCR technique was performed to identify the species of mink authenticity. The kit was effective after 20 repeated freeze\u2013thaw cycles and it could be stored at \u201320\u2009\u00b0C for 1 year. The sensitivity showed that a concentration as low as 0.1\u2009ng/\u03bcL still can amplify the target band. The specificity test confirmed that the kit was 100% specific. The kit proved to be effective, stable, and reliable for extraction of efficient contents of the genomic DNA and routine analysis of Chinese mink source composition from meat products.Species authentication of meat product origins has become an important subject for ensuring the health of consumers. Based on the cytochrome Therefore, food safety in meat, and meat products, is becoming increasingly important. Of all the food safety problems, the most important is food fraud, which is adulteration or substitution of expensive meat with an inferior meat. Recent years have seen an increase in the number of fraudulent cases involving deadly adulterants and legal prosecution for food fraud gene in mitochondrial DNA (mt DNA) to amply the specific fragment , mutton (QC-AN-002), pork (QC-AN-003), horse (QC-AN-004), rabbit (QC-AN-005), deer (QC-AN-006), duck (QC-AN-007), chicken (QC-AN-011), fox (QC-AN-013), and martes (QC-AN-014) were purchased from the National Center for Standard Reference Material, China. They were used as reference samples in the study and certified by Jilin Food Research Institute, National Institute for Food and Drug Control, China. A total number of 11 commercial samples consisting of beef, mutton, pork, chicken, duck, deer, horse, rabbit, and fox in addition to dog and donkey meat were purchased from a local supermarket and from randomly selected farmers\u2019 markets in the region. All animals\u2019 tissues in the study were carried out to comply with the Animal Research: Reporting In Vivo Experiments (ARRIVE) guidelines .One gram of each fresh sample was cut into small pieces, and dry samples (mass \u22650.1\u2009g) were washed, air-dried, cut into pieces, and stored in a 2-mL tube. The extraction protocol was done as previously described , a complex of phenols, was added to the extraction buffer to improve the extraction and purity of nucleic acids. The results showed that the DNA extracted was stable and sufficient in quantity. The design of mtDNA-specific primers is another key used to identify mink-derived components. The specific primers in the kit are The mink component DNA detection kit has good specificity, repeatability, sensitivity, and stability. The kit is highly sensitive and suitable for the detection of fresh meat and processed mixed materials. The mink source components DNA detection kit, for use in meat product quality assays, has a wide range of potential applications."} +{"text": "Drug repositioning (o repurposing) has become one of the most popular and successful strategies to reduce failures typically associated with drug discovery . It invocomputational and experimental approaches in the research area of repurposing natural products, including studies on, but not limited to, the identification of new biological targets of natural compounds, the discovery of bioactive natural and/or nature-inspired compounds by in silico screening, their isolation and characterization, de-replication of natural extracts, analysis of structure\u2013activity relationships of natural bioactive compounds. In light of all the above, this Special Issue will focus on Repositioning Natural Products in Drug Discovery\u201d meeting held at the University of Modena on Jan 17th, 2020 (http://www.mmddlab.unimore.it/site/home/rnpdd-meeting.html).Overall, this collection will provide a multi-disciplinary view of how different research fields can interact in either the discovery or the repurposing of bioactive nature-inspired compounds. This issue is connected to the \u201c"} +{"text": "When Dublin City University introduced the Age-Friendly University (AFU) principles in 2012, the Centre on Aging at the University of Manitoba had been fully immersed in age-friendly cities research, so it was a natural for it to champion the University becoming more age-friendly. As a university-wide research centre, at a research intensive university, the Centre on Aging has built on its strengths to advance the AFU movement. Related to its goals on research, knowledge mobilization, training, and partnerships, it has been able to work with its many Research Affiliates, students and community/university partners to tackle new AFU initiatives. Recent activities have included: an awareness raising workshop/showcase, discussions with Research Affiliates about potential new degree course offerings, an environmental scan of post-secondary inter-generational possibilities, a home-sharing project, workshop development on older learners in the classroom, and provincial community workshops and consultations on communicating about aging and healthy aging. Part of a symposium sponsored by Directors of Aging Centers Interest Group."} +{"text": "Besides general information including Q&As, daily case counts, and maps with disease distribution, examples of latest updates comprise: Resource estimation for contact tracing, quarantine and monitoring activities for COVID-19 cases in the EU/EEA, Guidance for wearing and removing personal protective equipment in healthcare settings for the care of patients with suspected or confirmed COVID-19 and Checklist for hospitals preparing for the reception and care of coronavirus 2019 (COVID-19) patients. ECDC also publishes regular risk assessments and the The European Centre for Disease Prevention and Control (ECDC) provides regularly updated information on coronavirus disease-2019 (COVID-19) relevant to Europe on a dedicated BoxOn 31 December 2019, a cluster of pneumonia cases of unknown aetiology was reported in Wuhan, Hubei Province, China. On 9 January 2020, China CDC reported a novel coronavirus as the causative agent of this outbreak, which is phylogenetically in the SARS-CoV clade. The disease associated to it is now referred to as novel coronavirus disease 2019 (COVID-19).As of 2 March 2020 at 08:00, more than 89,068 cases of COVID-19 have been reported worldwide, mainly in China and from all Chinese provinces; of these cases, around 9,000 cases were reported from other countries. As of 2 March, 66 countries have reported cases.In the EU/EEA, the UK, San Marino, Monaco and Switzerland, 2,199 cases have been reported as of 2 March. Among these cases, 38 have died. Italy represents 75% of the cases and 92% of the fatalities (n=35).Updates on the epidemiology of COVID-19 can be found on ECDC\u2019s website.COVID-19 is caused by a contagious newly identified virus. There are no therapeutics and vaccines available and there is presumably no pre-existing immunity in the population. Symptoms of COVID-19 range from no symptoms (asymptomatic) to severe pneumonia and can lead to death. The evidence from analyses of cases to date is that COVID-19 infection causes mild disease (i.e. non-pneumonia or mild pneumonia) in about 80% of cases and most cases recover, 14% have more severe disease and 6% experience critical illness. The great majority of the most severe illnesses and deaths have occurred among the elderly and those with other chronic underlying conditions.The risk associated with COVID-19 infection for people in the EU/EEA and UK is currently considered to be moderate to high, based on the probability of transmission and the impact of the disease. Based on the observed epidemiologic characteristics, everyone in the population is assumed to be susceptible, although there may be risk factors increasing susceptibility. The virus spreads rapidly, and can have an enormous public health impact with substantial fatal outcomes in high-risk groups and economic and societal disruption.Evidence from studies on influenza, and from recent experience in China, suggest that non-pharmaceutical interventions reduce transmission. Therefore, it is of paramount importance that measures that are appropriate and proportionate to each phase of the epidemic are immediately put in place to interrupt human to-human transmission chains, prevent further spread, reduce the intensity of the epidemic and slow down the increase in cases. Such measures should be coordinated at the EU level. This will ultimately reduce COVID-19 illness, save lives and minimise the socio-economic impact. Delaying transmission or decreasing the peak of the outbreak is crucial to allow healthcare systems to prepare and cope with an increased influx of patients.In addition, such a strategic approach based on rigorous application of these measures will allow more time for the testing of therapeutics and vaccine development. The different phases of the epidemic, e.g. from situations with no reported cases, sporadic cases or multiple introductions, local clusters of cases, to widespread sustained transmission, are referred to as scenarios in this document. Current epidemiology suggests scenario 1 (see main text for description) for EU/EEA level, which may be rapidly evolving to scenario 2. The options to be considered by national authorities for response appropriate to each scenario of the epidemic are described in detail under the dedicated section and include:Immediate activation of national emergency response mechanisms and pandemic preparedness plans to ensure containment and mitigation of COVID-19 with non-pharmaceutical public health measures.Ensuring the general public is aware of the seriousness of COVID-19. A high degree of population understanding, community engagement and acceptance of the measures put in place are key in preventing further spread.Implementation of protocols for COVID-19 laboratory testing, diagnosis, surveillance and treatment.Enhancement of surveillance, epidemiological investigation, close contact tracing, management of close contacts, immediate case detection and isolation.Implementation of social distancing to interrupt the chains of transmission.Adapted risk communication and provision of adequate personal protective equipment for healthcare workers and rigorous application of infection prevention and control measures in healthcare facilities.Provision of adequate healthcare capacity to isolate, support and actively treat patients.What is new in this update?Updated number of cases in China, in EU/EEA and globallyFindings on disease and transmissibility from recent studiesRisk associated with COVID-19 for people from the EU/EEA and the UK resident/travelling in areas with no cases, or multiple imported cases, or limited local transmissionRisk to the healthcare systems in the EU/EEA and the UKRisk of widespread and sustained transmission in the EU/EEA and UK in the coming weeksOptions for preparedness and response; including a proposed change in the case definition and the integration of testing for COVID-19 in surveillance systems for influenza surveillance (ARI/ILI) and severe acute respiratory infections.https://www.ecdc.europa.eu/sites/default/files/documents/RRA-outbreak-novel-coronavirus-disease-2019-increase-transmission-globally-COVID-19.pdfSource: European Centre for Disease Prevention and Control (ECDC). Outbreak of novel coronavirus disease 2019 (COVID-19): increased transmission globally \u2013 fifth update. ECDC: Stockholm; 2 March 2020. Available from: ARI: acute respiratory infection; COVID: coronavirus disease; ECDC: European Centre for Disease Prevention and Control; EU/EEA: European Union/European Economic Area; ILI: influenza-like-illness; SARS-CoV: severe acute respiratory syndrome coronavirus; UK: United Kingdom."} +{"text": "Yunfeng Fu was added as an author to thank him for the experimental and technical guidance provided in the implementation of previous projects, albeit without his authorization; furthermore, the authors now recognize that the inclusion of Dr Fu as an author did not meet with the criteria proposed by the International Committee of Medical Journal Editors (IJCME) for authorship on scientific articles. All the original authors (including Yunfeng Fu) agree with the revision made to the authorship on this paper, and the authors remaining on the paper apologize for any inconvenience caused. Therefore, the revised authors\u2019 names and affiliations in this paper (including the revised Corresponding Author details) are as follows:1, GONG-XIANG LI1 and LINGLI QUAN2SHU-HUI XIAO1Department of Clinical Laboratory Medicine, The People's Hospital of Jinyi, Jinyi, Shanxi 710003; 2The First Department of Respiratory of Central Hospital of Zhuzhou, Zhuzhou, Hunan 412000, P.R. ChinaCorrespondence to: Dr Lingli Quan, The First Department of Respiratory of Central Hospital of Zhuzhou, 116 South Changjiang Road, Zhuzhou, Hunan 412000, P.R. Chinajing1969wang@sina.comE-mail:"} +{"text": "Nature Communications 10.1038/s41467-019-10756-2, published online 02 July 2019.Correction to: https://www.synapse.org/#!Synapse:syn22213200]. This has now been corrected in the PDF and HTML versions of the paper. Additionally, Yiming Yang was incorrectly associated with \u2018Department of Biological Sciences, Columbia University, New York, NY, 10027, USA\u2019 and Naomi Habib was incorrectly associated with \u2018Center for Immunology and Inflammatory Diseases, Division of Rheumatology, Allergy, and Immunology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, 02129, USA\u2019 in the HTML version of the paper. The PDF was correct at the time of publication. This has now been corrected in the HTML version of the paper.In the original version of this article, the raw human sequencing data was available at the RADC Resource Sharing Hub through a data use agreement. This data can now be accessed at the Synapse AMP-AD Knowledge Portal ["} +{"text": "Background: The association between attention-deficit hypersensitivity disorder (ADHD) and the risk of developing colorectal cancer (CRC) is, as yet, to be investigated, and thus, we have conducted this nationwide, cohort study to examine the association in patients from Taiwan.Methods: In this study, 798 individuals with newly diagnosed ADHD and 2,394 (1:3) age-, gender-, and index year- matched controls without ADHD were enrolled, between 2000 and 2013, from the Longitudinal Health Insurance Database, a subset of the National Health Insurance Research Database in Taiwan. The cumulative incidence of CRC was assessed in each cohort by the Kaplan\u2013Meier method. The multivariate Cox proportional hazards model was used to estimate the crude, and the adjusted hazards ratios (HRs) with 95% confidence intervals (CIs), was conducted to estimate the association between ADHD and CRC.Results: The Kaplan\u2013Meier analysis revealed that the cumulative incidence of CRC was significantly higher in patients with ADHD than in those without it . After adjustments for age, gender, comorbidities, and other covariates, the ADHD group was associated with an increased risk of CRC in comparison to the non-ADHD group . In addition, the usage of methylphenidate was not associated with the risk of developing CRC in patients with ADHD.Conclusion: This retrospective cohort study depicts the evidence that ADHD was associated with the increased risk of CRC. Further studies are needed to confirm the association and the underlying mechanisms. Colorectal cancer (CRC) is a major health challenge, representing the most common cancer and the third most frequent cause of cancer-related deaths in Taiwan . Some reAttention-deficit/hyperactivity disorder (ADHD) is one of the most common pediatric neurodevelopmental and neurobehavioral disorders with a worldwide average prevalence estimated at 5% in children and 3.4\u20134.4% in adults, which results in attention deficit, hyperactivity, and increased impulsivity , and up ADHD have bidirectional relations , for exaThe NHIRD was established in 1995, and as of June 2009, included contracts with 97% of the medical providers with ~23 million people in the program, or more than 99% of the entire population in Taiwan . The detThis was a retrospective cohort study. Patients with ADHD between January 2000, and December 2013, were selected from the LHID and categorized according to the ICD as 314. All diagnoses of ADHD were made by board-certified specialists such as psychiatrists, pediatricians, neurologists, or physiatrists with specialty in child and adolescent development. The subject selection process is as presented in We followed up both cohorts from January 1, 2000, until the date of CRC diagnosis , withdrawal from the NHI, or the end of 2013.The covariates included the age, gender, monthly insured premiums, comorbidities, locations, urbanization levels of residence, and the levels of hospitals for medical help. In the analysis, since the north Taiwan is the center of politics and economics, in the country. In addition, most of the healthcare resources, for example, 12 medical centers among total 23, located in the northern Taiwan. Therefore, the northern Taiwan was listed as the reference in our study for the locations.We noticed the covariates that were potential confounders in the association between ADHD and CRC including age, gender, and the underlying chronic diseases related to the risk of developing CRC. Those chronic diseases, which were taken into account, included chronic obstructive pulmonary disease (COPD) , diabetes mellitus (DM) (ICD-9-CM code: 250), coronary artery disease , hypertension , hypercholesterolemia , alcohol-related diseases , peptic ulcer ; liver cirrhosis and chronic hepatitis (ICD-9-CM code: 571), inflammatory bowel disease (ICD-9-CM codes: 555 and 556), and psychiatric comorbidities such as oppositional defiant disorder , conduct disorder , autism spectrum disorder ; tic disorder (ICD-9-CM code: 307.2); intellectual disabilities ; anxiety (ICD-9-CM code: 300); depression ; and bipolar disorder . All the diagnosis of the psychiatric disorders was conducted by the board-certified psychiatrists, pediatricians, neurologists, and physiatrists.t-tests when appropriate. In addition, we used the Kaplan-Meier method to estimate the cumulative incidence of CRC in the study cohorts and performed the log-rank test to compare the difference between these two curves. We computed the hazard ratios (HRs) presented together with 95% confidence intervals (CIs) using the Cox proportional hazards models after adjusting for the potential confounders mentioned above. All the confounders, as covariates and comorbidities, including the psychiatric diagnoses, were calculated separately. All analyses were performed using the SAS version 9.4 and the statistical significance was set to 0.05 in the 2-tailed tests.We examined the descriptive statistics of the demographic characteristics and baseline comorbidities between the exposed and non-exposed cohorts by conducting chi-square tests or Student's-A total of 3,192 patients were enrolled in this study, including 798 adult patients with ADHD and 2,394 patients in the non-ADHD control cohort. The age, gender, monthly insured premiums, comorbidities, locations and urbanization levels of residence, and the levels of hospitals for medical help are as summarized in p < 0.001) (p = 0.03).The Kaplan-Meier analysis for the cumulative incidence of CRC in the ADHD and non-ADHD cohorts with the log-rank test revealed a significant difference over the 13-year follow-up period (< 0.001) . In the p < 0.001), and after adjusting for age, gender, comorbidities, geographical area of residence, urbanization level of residence, and monthly income, the adjusted HR was 3.458 . For the participants older than 50 years, in both the ADHD and non-ADHD cohort, the risk of CRC was 6.887 , in comparison to the subgroup aged from 20 to 49. In contrast, the ADHD group was uniformly associated with an increased risk of CRC for all the factors.In p < 0.001), when compared with the non-ADHD control group. The Kaplan-Meier analysis revealed that the study subjects had a significantly higher 14-year cumulative incidence of ADHD than the controls. Furthermore, the ADHD-cohorts older than 50 years had a nearly 6.9-fold increased risk of CRC . This study revealed that patients with ADHD had a nearly 3.5-fold risk of CRC, and this report could be a reminder for the clinicians who care the patients of ADHD in the follow-up. To the best of our knowledge, this is the first nationwide, population-based cohort on the topic of the association between ADHD and the risk of developing CRC.In this study, we examined the association between ADHD and the risk of CRC. After adjusting the covariates, the adjusted HR was 3.458 for the ADHD group claim database system; therefore, some registry bias may have been involved in the calculation of the CRC risk. In addition, the family history of colorectal cancer is present, up to one third of patients . The lacThis retrospective cohort study provided evidence of a nearly 3.5-fold increased risk of CRC in ADHD. The results of this study could serve as a reminder for the clinicians who care for the patients of ADHD in the follow-up. Further prospective studies are necessary for confirming our findings, we therefore recommend meticulous evaluation and aggressive risk reduction for CRC for the patients with ADHD.https://nhird.nhri.org.tw/en/index.html).The datasets analyzed in this article are not publicly available. Requests to access the datasets should be directed to Data are available from the National Health Insurance Research Database (NHIRD) published by the Taiwan National Health Insurance (NHI) Administration. Due to legal restrictions imposed by the government of Taiwan in relation to the \u201cPersonal Information Protection Act\u201d, data cannot be made publicly available. Requests for data can be sent as a formal proposal to the NHIRD . Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.J-MH, C-CL, N-ST, and W-CC conceived, planned, and conducted this study. C-CL, J-MH, N-ST, C-HC, and W-CC contributed to the data analysis and interpretation. J-MH, T-CL, C-HC, C-YC, P-KC, C-AS, and C-WH contributed to this data interpretation. C-CL wrote the first draft. J-MH has played major role, in this revision, in the concept, data interpretation, data analysis, and the re-writing of this manuscript. N-ST and W-CC conducted the critical revisions of this article. All authors approved this manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Nature Communications 10.1038/s41467-020-16266-w, published online 15 May 2020.Correction to: The original version of this Article contained errors in the author affiliations.Ye-Liang Wang was incorrectly associated with the School of Materials Science and Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.Wei Ji was incorrectly associated with the Songshan Lake Materials Laboratory, 523808 Dongguan, China.These have now been corrected in both the PDF and HTML versions of the Article."} +{"text": "Nature Communications 10.1038/s41467-020-15816-6, published online 24 April 2020.Correction to: The original version of this article contained an error in the author affiliations. The affiliation of Eran Halperin with Institute for Precision Health, School of Medicine, UCLA, Los Angeles, CA 90095, USA was inadvertently omitted. This has now been corrected in both the PDF and HTML versions of the article."} +{"text": "Acer tegmentosum Maxim. The plastome of A. tegmentosum was 156,435\u2009bp in length and included both large and small single-copy regions, which were separated by a pair of identical inverted repeats . The plastome contained 77 unique protein-coding genes, 30 tRNA genes, and four rRNA genes. In addition, the gene order and organization of the A. tegmentosum plastome were consistent with those of plastomes from other members of the Sapindaceae, and the overall GC content of the plastome was 37.8%. A phylogenetic tree that was based on 76 protein-coding genes demonstrated a sister relationship within genus Acer.The aim of the present study was to sequence and analyze the complete plastid genome (i.e. plastome) of Acer L. (Sapindaceae) species are some of the most important trees in the Northern Hemisphere, particularly in the temperate regions of eastern Asia, eastern North America, and Europe will contribute to the development of a protection strategy for the species.asin Kim . The speasin Kim . TherefoAcer buergerianum (GenBank accession number KX098452) as a reference sequence. The sequenced fragments were assembled using Geneious R10 and BLAST searches. The tRNA genes identified using Geneious were validated using the web-based tool tRNAScan-SE , and genomic DNA was isolated from fresh leaves using a Plasmid SV mini kit . Extracted DNA were stored in the Plant DNA Bank of the National Institute of Forest Science . Whole genome sequencing was performed using the Ion Torrent sequencing platform and filtered sequences were assembled using the plastome of A. tegmentosum was double-stranded, circular, and 156,435\u2009bp long, with two inverted repeat regions that were separated by large single-copy and small single-copy regions (GenBank accession number MK942342). The plastome contained 128 genes, including 83 protein-coding genes, 37 tRNA genes, and eight rRNA genes, and six, seven, and four of the protein-coding, tRNA, and rRNA genes, respectively, were duplicated in the IRs. In addition, the overall GC content of the plastome was 37.8% , and the monophyly of the genus was well-supported , with A. tegmentosum placed as sister to A. morrisonense (A. tegmentosum plastome described here may contribute to a better understanding of the evolution of Acer.The plastome of isonense . The A."} +{"text": "Since its inception, RNA sequencing (RNA-seq) has become the most effective way to study gene expression. After more than a decade of development, numerous RNA-seq datasets have been created, and the full utilization of these datasets has emerged as a major issue. In this study, we built a comprehensive database named Grape-RNA, which is focused on the collection, evaluation, treatment, and data sharing of grape RNA-seq datasets. This database contains 1529 RNA-seq samples, 112 microRNA samples from the public platform, and 485 RNA-seq in-house datasets sequenced by our lab. We classified these data into 25 conditions and provide the sample information, cleaned raw data, expression level, assembled unigenes, useful tools, and other relevant information to the users. Thus, this study provides data and tools that should be beneficial for researchers by allowing them to easily use the RNA-seq. The provided information can greatly contribute to grape breeding and genomic and biological research. This study may improve the usage of RNA-seq. Gene expression is one of the most important processes in life activities, playing a vital role in the development and environmental adaptation of living things. The study of gene expression is fundamental for functional genomics research, especially in the post-genomic era, which began with the completion of the Human Genome Project in 2001 and sample ID (GWGT), are necessary to search for the expression level of RNA-seq. The expression level of the required genes will be listed in table format on the result page. For example, if we want to know the expression level of several genes during heat stress on grape berry, the gene ID and sample ID of 48 samples related to heat stress on grape berry should be input, and the expression level of these genes will be searched and provided on the results page. Users can also download the expression information file from a download link. This expression file can be used as the input files for the heatmap tools. Expression level of microRNA only requires the sample ID, and all microRNA information of this sample will be listed in the result page. In addition to these three search tools, we provide the search tools of GO, KEGG, NR annotation, and some related papers. These tools can supply multiple types of functional annotation of the targeted genes and give some guidance to the users, and also can do benefit to other works, such as RNA-seq analysis and gene identification.Several searching tools were developed in this database (http://www.grapeworld.cn/gt/tools.html). Some of these tools are BLAST, BLAT, GO and KEGG annotation, signal peptide identification, transmembrane finder, RT-qPCR primer design, sequence tools, and microRNA regulatory network. These tools were developed based on all existing annotation versions of 8X and 12X grape genome, and supplied the information of similar sequences, high quality primers, or the information of signal peptide, transmembrane domain, and microRNA regulation. These tools can help researchers avoid the need for complex programming languages and a huge number of computations, making the undertaking of related studies easier and more efficient. In addition, 1529 high-quality samples were used in the co-expression analysis. This analysis was based on the IGGPv2.1 annotation version and processed by WGCNA (v1.63) [Grape-RNA includes abundant tools to make the research of related studies more convenient ( (v1.63) . The thr (v1.63) . For exahttp://www.grapeworld.cn/gt/help.html) to allow users to quickly begin using this database. We provide detailed instructions with a screenshot for each section. All introduction files are in PDF and can be accessed freely through the network.In Grape-RNA, we provide a help page and can be accessed worldwide at any time. The database will be updated when enough new sequenced RNA-seq data have been collected or a new analysis method emerges.Grape-RNA is a part of Grapeworld (Grape-RNA is currently the most comprehensive and versatile grape RNA-seq database and provides a user-friendly interface. Compared with VitisNet and VESP"} +{"text": "In this work, surface/interface effects for pull-in voltage and viscous fluid velocity effects on the dimensionless natural frequency of fluid-conveying multiwalled piezoelectric nanosensors (FC-MWPENSs) based on cylindrical nanoshells is investigated using the Gurtin\u2013Murdoch surface/interface theory. The nanosensor is embedded in a viscoelastic foundation and subjected to nonlinear van der Waals and electrostatic forces. Hamilton\u2019s principle is used to derive the governing and boundary conditions and is also the assumed mode method used for changing the partial differential equations into ordinary differential equations. The influences of the surface/interface effect, such as Lame\u2019s constants, residual stress, piezoelectric constants and mass density, are considered for analysis of the dimensionless natural frequency with respect to the viscous fluid velocity and pull-in voltage of the FC-MWPENSs. Nanomechanical sensors and resonators, especially when combined with piezoelectric materials, are widely used in modern engineering, which encompasses numerous, diverse fields of science and technology, pharmaceutical, agricultural, environmental, advanced materials, chemical science, physics, electronics, information technology, biomedical and medical fields \u201310. Due Many studies have been carried out on the vibration and stability analysis of nanostructures with some reviews given as follows. Strozzi and Pellicano investigated the vibration analysis of triple-walled carbon nanotubes (TWNTs) subjected to the interlayer van der Waals (vdW) force in the framework of the Sanders\u2013Koiter shell theory . Also, bI,S, \u00b5I,S), residual stress piezoelectric constants and mass density are studied for analysis of dimensionless natural frequency with respect to viscous fluid velocity and pull-in voltage of fluid-conveying multiwalled piezoelectric nanosensors (FC-MWPENSs) subjected to direct electrostatic DC voltage with nonlinear excitation, nonlinear van der Waals force and viscoelastic foundation. As a guide to the reader, all notation and symbols are presented in To the best knowledge of the author, the surface/interface effect on pull-in voltage, viscous fluid velocity effects and dimensionless natural frequency (DNF) of multiwalled piezoelectric nanosensors conveying viscous fluid has not yet been studied. In the present study, the effect of surface/interface parameters such as Lame\u2019s constants for the other inner wall layers. All of the physical and geometrical properties of the mentioned nanostructures for single-walled piezoelectric nanoresonators can be seen in work done by Hashemi Kachapi et al. in reference [A schematic diagram of a multiwalled piezoelectric nanosensor with an embedded fluid-conveying inner layer, two piezoelectric layers, and a viscoelastic foundation medium in the outer layer is shown in eference ,20.The governing equations and the solution procedure can be found in I,S, \u00b5I,S), residual stress piezoelectric constants and mass density , are investigated for analysis of the dimensionless natural frequency with respect to viscous fluid velocity and pull-in voltage In order to simplify the presentation, CC, SS and CS represent the clamped edge, simply supported edge and clamped-simply supported edge, respectively. The material properties of the different layers of aluminum (Al) nanoshell and piezoelectric layers (PZT-4) are shown in A verification study is investigated in work by Hashemi Kachapi et al. \u201320 with The other bulk and surface geometrical parameters of FC-MWPENS are shown in R = Rout; for double-walled PENS (DWPENS): R1 = Rin, R2 = Rout; and for triple-walled PENS (TWPENS): R1 = Rin, R2 = Rmid, R3 = Rout.The value of the mid-surface radius for different PENS are presented as following: for single-walled PENS (SWPENS): I,S, \u00b5I,S), residual stress piezoelectric constants and mass density , of fluid-conveying multiwalled piezoelectric nanosensors are studied for analysis of the dimensionless natural frequency (DNF) with respect to viscous fluid velocity and pull-in voltage For this work, the material and geometrical parameters in Tables 2\u20134 are used. In all of the following results of the analysis of DNF on viscous fluid velocity and pull-in voltage respectively, the values of = 5 and = 0.1 are used.In this section, the effect of surface/interface parameters, such as Lame\u2019s constants , the inertia of the shell is increased and its stiffness is reduced, which leads to a decreased DNF compared to the case without surface/interface effects. Also, with decreasing surface/interface density (case 2), the inertia of the system is increased, and with increasing stiffness, DNF increases compared to the case without surface/interface effects.It is observed that for all modes, the DNF decreases when the and pull-in voltage In all of the following results, the lower surface/interface density (case 2) is used in the analysis of DNF on viscous fluid velocity and direct pull-in DC voltage on the pull-in instability analysis on the DNF of FC-MWPENS are presented in The effects of viscous fluid velocity and for viscous fluid velocity and pull-in instability analysis on DNF of FC-MWPENS. It is clear that the increasing surface/interface Lame\u2019s constants \u03bbI,S, due to increasing FC-MWPENS stiffness, DNF and critical fluid velocity increase and pull-in voltage in \u03bbI,S = 0 and \u03bbI,S = \u22122 has a maximum and a minimum value. and for viscous fluid velocity and pull-in instability analysis on DNF of FC-MWPENS are presented in I,S, it is clear that by increasing both surface/interface Lame\u2019s constants \u00b5I,S, due to increasing FC-MWPENS stiffness, the DNF and also the critical fluid velocity and pull-in voltage increase.The effects of different surface and interface Lame\u2019s constants, and for viscous fluid velocity and pull-in instability analysis on DNF of FC-MWPENS. As can be seen in the analysis of DNF, increasing the surface/interface residual stress leads to increasing FC-MWPENS stiffness, and as a result, the DNF and pull-in voltage increase and critical fluid velocity decreases. and for viscous fluid velocity and pull-in instability analysis on DNF of FC-MWPENS is presented in and leads to increasing FC-MWPENS stiffness, and as a result, the DNF, critical fluid velocity and pull-in voltage increase.The effect of surface piezoelectricity constants and for viscous fluid velocity and pull-in instability analysis on DNF of FC-MWPENS. As it can be seen, with an increasing surface/interface mass density \u03c1I,S, due to increasing FC-MWPENS stiffness, the DNF significantly increases and also the critical fluid velocity and pull-in voltage slightly decrease. and pull-in instability analysis on DNF of SS FC-MWPENS are presented. It can be seen that by ignoring the surface/interface density \u03c1I,S, the inertia of the system will greatly decrease and due to increasing FC-MWPENS stiffness, the system will have a maximum DNF compared to other cases. Also when the surface/interface effects are not taken into account, due to the decreasing nanoshell stiffness, it has a lower DNF than the case with all surface/interface effects. In the cases without all surface/interface effects, the critical fluid velocity and also the pull-in voltage reach zero sooner than the rest of the parameters. In the cases without surface/interface density \u03c1I,S and with all surface/interface effects, the pull-in voltage and critical fluid velocity reach zero later than the rest of the parameters and the system loses its stability due to the divergence via a pitchfork bifurcation.In and pull-in voltage The piezoelectric nanosensor is simultaneously subjected to direct electrostatic DC voltage with nonlinear excitation, nonlinear van der Waals forces and a viscoelastic foundation. For this purpose, Hamilton\u2019s principles, the assumed mode method combined with Lagrange\u2013Euler\u2019s equations are used. The results demonstrated that in the case of higher (lower) surface/interface densities, the inertia of the shell is increased (decreased) and its stiffness is reduced (increased), which leads to a decreasing (increasing) natural frequency compared to the case of without surface/interface effects. Also, by increasing both surface/interface Lame\u2019s constants, \u03bbI,S and \u00b5I,S, and the negative surface piezoelectricity constants, and due to the increasing FC-MWPENS stiffness, the DNF and also the critical fluid velocity and pull-in voltage increase. In addition, in the analysis of DNF, it was found that increasing the surface/interface residual stress leads to increasing FC-MWPENS stiffness, and as a result, the DNF and pull-in voltage increase and critical fluid velocity decreases. Increasing the surface/interface mass density, \u03c1I,S, due to increasing FC-MWPENS stiffness, it was found that the DNF significantly increases and also the critical fluid velocity and pull-in voltage slightly decrease. Finally, by ignoring the surface/interface density, \u03c1I,S, the system will have a maximum DNF compared to other cases. In cases without all surface/interface effects, the critical fluid velocity and also the pull-in voltage reached zero sooner than the rest of the parameters. In the cases without surface/interface density \u03c1I,S and with all surface/interface effects, the pull-in voltage and critical fluid velocity reach zero later than the rest of the parameters.In the current study, the effect of the surface/interface parameters of a fluid-conveying multiwalled piezoelectric nanosensor are studied for analysis of the dimensionless natural frequency with respect to viscous fluid velocity File 1Subsections of \u201cMathematical Formulation\u201d as well as an \u201cAppendix\u201d section.File 2MATLAB program code for the current paper.In the ZIP file, all programs written in MATLAB software are presented for the results of the article, which is not included in the article due to the large volume of the program."} +{"text": "This data article includes information on institutional data at a large public research university in Southern California. In particular, data on undergraduate student enrollments in online and face-to-face courses during summer terms from 2014 to 2017 cumulating in 72,441 course enrollments from 23,610 undergraduate students in 433 courses is provided. This data includes additional information on the statistical models examining factors influencing student enrollment by course modality and the associations of course modality with course grades. This includes descriptive data and data derived from multi-level logistic regression analyses and multi-way fixed effects linear regression analyses. This data article is associated with the article \u201cEffects of course modality in summer session: Enrollment patterns and student performance in face-to-face and online classes\u201d [1]. The dat22.1Students enrolled in undergraduate courses at in the 2014 to 2017 summer terms were included in this data. This data includes enrollments from degree-seeking undergraduate students in lecture courses that were graded on a letter-grade scale (in contrast to a pass/no-pass distinction) and awarded at least four credit points. Data was provided by the Teaching and Learning Research Center and was compiled from a variety of sources on campus including: Office of Institutional Research, Office of the Registrar, Office of Undergraduate Admissions, Office of Institutional Technology, and the Office of Financial Aid and Scholarships. This project was supported by the UC Irvine Institutional Review Board. The data consisted of student-level demographic, performance, and college career information, as well as course-level information.Student-level demographic information included gender , racial/ethnic background , first-generation college student status , low-income status , English language learner status , and whether or not the student is a California resident. Student performance indicators include, standardized admission test score and the Scholastic Aptitude Test (SAT) scores) and current college grade point averages. College career characteristics included students' years of enrollment in the institution, transfer student status, the number of online courses taken in college, and whether students repeated courses.Course-level data includes course grades, course code, department, year and term the course was offered, the number of students enrolled in a course, course modality , and unique instructor identification information.2.2meqrlogit syntax in Stata 15 [The following tables describe descriptive information of (a) all course enrollments, (b) all face-to-face course enrollments, and (c) all online course enrollments. This information is provided for both the full sample and a restricted sample that only includes courses that were offered as both online and face-to-face courses in the same term , Table 32.3xtreg syntax in Stata 15 [The following table describes descriptive information for (a) all course enrollments, (b) all course enrollments of low-income students, (c) all course enrollments of first-generation college students, and (d) all course enrollments of students in the lower high school performance subgroup. This information is displayed separately by online and face-to-face course enrollments . The desStata 15 . Please"} +{"text": "Nature Communications 10.1038/s41467-020-14966-x, published online 4 March 2020.Correction to: The original version of this Article contained an error in the author affiliations.Affiliation 2 incorrectly read \u2018King Abdulaziz University, University of Jeddah, Jeddah, Saudi Arabia\u2019 instead of the correct \u2018University of Jeddah, College of Science, Department of Biology, Jeddah, Saudi Arabia\u2019.In addition the original version of this Article did not acknowledge Hebah Sindi as a corresponding author. This has now been corrected in both the PDF and HTML versions of the Article.These errors have now been corrected in both the PDF and HTML versions of the Article."} +{"text": "With the immersive growth of the Internet of Things (IoT) and real-time adaptability, quality of life for people is improving. IoT applications are diverse in nature and one crucial aspect of it is multimedia sensors and devices. These IoT multimedia devices form the Internet of Multimedia Things (IoMT). It generates a massive volume of data with different characteristics and requirements than the IoT. The real-time deployment scenarios vary from smart traffic monitoring to smart hospitals. Hence, Timely delivery of IoMT data and decision making is critical as it directly involves the safety of human beings. In this paper, we present a brief overview of IoMT and future research directions. Afterward, we provide an overview of the accepted articles in our special issue on the IoMT: Opportunities, Challenges, and Solutions. Internet of Things (IoT) devices has limited memory and processing capabilities . Hence, Internet of Multimedia Things (IoMT) devices are different from IoT devices. It requires bigger memory, higher computational power, and more power-hungry with higher bandwidth . Figure IoT characteristics support multimedia communications; however, multimedia applications are bandwidth-hungry and delay-sensitive. The rapid growth of multimedia traffic in IoT has led the way to innovating new techniques to meet its requirements. IoMT devices require higher bandwidth, bigger memory, and faster computational resources to process data. Typical communications include multipoint-to-point and multipoint-to-multipoint scenarios. Real-world multimedia applications include emergency response systems, traffic monitoring, crime inspection, smart cities, smart homes, smart hospitals, smart agriculture, surveillance systems, Internet of bodies (IoB), and Industrial IoT (IIoT). Dynamic networks, heterogeneous devices and data, strict QoS, and delay sensitivity and reliability requirements over resource-constrained IoMT pose humongous challenges for multimedia communication in IoT. Network-on-chip architecture ,19 is onThe rest of the paper is organized as follows. Molecular communication exploits the transmission and reception of information encoded in molecules. Molecular communications have the potential of becoming the main technology for the execution of advanced medical solutions. The key research challenges in the molecular communication are the interoperability between molecular communication and the other networks, energy-efficient models and protocols for molecular communication, and implantation of reliability in the molecular communication.Billions of resource constraint devices will be connected in the IoT. The available spectrum is far from enough to support IoT communication systems. Optimal resource allocation for critical multimedia traffic is a key challenge for IoMT. The use of artificial intelligence can improve the energy-efficient resource allocation in IoMT.Device-to-device (D2D) communication in LTE-A will establish direct communication with the device in its communication range. Potential advantages of D2D communication are increased network spectral efficiency, energy efficiency, reduced transmission delay, traffic offloaded base station, and less congestion in the cellular core network. IoMT can take the benefits of the advantages provided by D2D communication. Interference, radio resource allocation, power control, and QoE improvement for cellular users are the key research areas in D2D communication for IoMT.Energy Efficient Operation and Protocols are the key requirement of IoMT. Many multimedia traffic sources in IoMT may rely on battery-powered sources with limited energy and/or may not be easily accessible for recharging purposes. Similar to WSNs/IoT, the energy-efficient operation, protocols design , and the need to optimize the network lifetime remains a critical challenge for IoMT. Based on the specific application and deployment environment, energy-efficient protocols can be designed for IoMT.The Internet of Multimedia Nano-Things (IoMNT) is defined as the interconnection of multimedia nano-devices with communication networks and the Internet. The potential applications of IoMNT are security, biomedical, defense, and industry. The main research challenges in IoMNT includes novel medium access control techniques, addressing schemes, neighbor discovery and routing schemes, QoS-aware cross-layer communication module and security solutions for the IoMNT.Multimedia-oriented IoT over vehicular networks is increasing drastically. Today, vehicles have the capability of supporting real-time acquisition and transmission of the multimedia traffic generated by the built-in IoT devices. However, due to high mobility, density, and random wireless channel conditions, the performance of the delivery of multimedia contents significantly reduces in vehicular networks. Rate adaptation, multimedia delivery over heterogeneous devices, robust video encoding, scalable, and timely delivery of multimedia contents are the key research challenges in IoMT over vehicular networks.The immense growth in multimedia traffic over the scarce licensed cellular spectrum has inspired to use unlicensed spectrum below 6 GHz for Long Term Evolution (LTE). However, Wi-Fi uses the same unlicensed band, and this gives rise to the issue of coexistence and fairness of two different technologies in the context of physical and link layer protocols. LTE in the Unlicensed (LTE-U) and LTE License Assisted Access (LTE-LAA) has been proposed in the literature for IoT system. The Third Generation Partnership (3GPP) has standardized LAA for industrial IoT. The coexistence mechanism of LAA follows Listen Before Talk (LBT), which is the same process of Wi-Fi system coexistence i.e., Carrier Sense Multiple Access (CSMA). LTE-U operates a carrier ON/OFF switch policy in duty cycles to maintain fairness in LTE and Wi-Fi transmissions. This mechanism causes spectrum inefficiency. Bajracharya et al. proposedWith the exponential growth of the IoT, the interaction of multiple physical devices is of extreme importance. These devices are often integrated using Radio Frequency Identification (RFID). The RFID automatically recognizes the object details by reading the physical objects. The system reader, which is equipped with a backend server, uses radio frequencies to communicate with the objects with RFID tags. It makes the practical usage of RFID very vast. Security is the most vital aspect of communication for authentication and securing private data. RFID-based security is beneficial in multiple ways, as RFID does not require a light source and line of sight scenario for communication. Hence RFID can be deployed to sensor monitoring, access control, real-time inventory, and security-aware management systems. However, due to limited computational and memory resources on an RFID tag, limited cryptographic operations can be applied. Therefore, an eavesdropper can forge and access the user\u2019s private data. David et al. in proposedWater is the soul of life and essential to the well-being of every person, economy, and the ecosystem on Earth. More than 70% of the Earth is surface is covered by oceans, and they are critical to maintaining the weather and temperature around the globe and providing a means of transportation. However, more than 90% of oceans are unexplored even to the extent that they are still unseen by humans. IoT paved the way to explore and collect the data by connecting different types of networks underwater. Such networks are known as the Internet of Underwater Technology (IoUT) is an emerging technology to support Underwater Sensors Networks (UWSNs) for exploration of undiscovered marine resources. UWSN using communication cables and sensors and maintenance cost is very high. Therefore, underwater wireless communication is proposed. However, UWSN wireless communication is challenging due to environment and propagation losses, which include high noise, Doppler spreading, path loss, multi-path signal propagation, and high power consumption. To overcome these issues, Faheem et al. in proposedUltra Wide band (UWB) features include higher bandwidth, and it is one of the viable technologies for IoMT applications. The integration of UWB in health critical IoT applications can provide an effective and reliable solution for the monitoring of patients. Ataxia patients suffer from abnormal movement, and that severely affects walking activities. The walking activities can be classified as a normal walk, difficulty walking in a straight line, walking with heavy steps, and forward bending walking. All of these walking patterns except normal walking shows abnormality. Zilani et al. proposedSecurity, privacy, and trust remain challenges in IoMT because of the openness and heterogeneity of IoMT. Access control is used to protect the confidentiality and integrity of constrained resources in IoMT services. It provides a solution to avoid any unauthorized access for multimedia applications in IoT services. However, due to increasing the number of users and multimedia services offered by the IoT platform, the access control system is becoming more and more multifaceted. Besides, access control policy evaluation reduces the performance of IoMT applications. Therefore, Meiping Liu et al. proposedIncreasing demand for data-intensive applications is growing users\u2019 data requirements exponentially. However, spectrum scarcity is the biggest hurdle in meeting users QoS. One of the viable solutions is to reuse and share the spectrum among the users to fulfill users demands without compromising the user\u2019s experience. Even though unlicensed spectrums are available for free, they are already overcrowded. Different network technologies such as 5G and Wi-Fi use different spectrum access mechanisms, and sharing the spectrum among them is a trivial task. To allow fair coexistence between 5G and Wi-Fi networks operating in the same spectrum, LBT is introduced to work in parallel with the CSMA for channel access. RL techniques can be adapted to make a spectrum access mechanism to learn network conditions itself and adapt to the network changes accordingly. Consequently, the network becomes sustainable and self-adaptive. Neto et al. in proposedSix papers in this SI presented state-of-the-art research trend in the area of IoMT opportunities, challenges, and solutions. The papers presented an interesting discussion and novel ideas for the readers. The guest editors would like to show appreciation to the authors and thank all the anonymous reviewers for providing constructive feedback to improve the overall quality of all the accepted papers. We would also like to thank sensors Editor in Chief Prof. Dr. Vittorio M.N. Passaro, Associate Editor in Chief of the IoT section Prof. Dr. Raffaele Bruno, and managing editor Missy Wu for the invaluable help and productive advice in finalizing this SI."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "While family engagement at the individual level of health care, such as families partnering with providers in decision-making about health care for an individual child has been well studied, family engagement in systems-level activities that impact the health services families and children receive has not. This note from the field presents a framework that describes the information and supports that help families partner with professionals and contribute to systems-level activities. Without attention to these components of family engagement, family presence and participation may be only token. We engaged an expert Family/Professional Workgroup whose members represented key constituencies and diverse geography, race/ethnicity, and areas of expertise; conducted a review of peer-reviewed publications and grey literature; and conducted a series of key informant interviews to identify best practices for supporting meaningful family engagement at the systems level. Based on an analysis of the findings, the authors identified four action-oriented domains of family engagement and key criteria that support and strengthen meaningful family engagement in systems-level initiatives. Child- and family-serving serving organizations can use this Family Engagement in Systems framework to support meaningful family engagement in the design of policies, practices, services, supports, quality improvement projects, research, and other systems-level activities. Report on Children with Special Health Care NeedsIn 1987, Surgeon General C. Everett Koop\u2019s Assure families and individuals are key partners in health care decision-making at all levels across the health care system and the services that support them, especially those who are vulnerable and medically underserved.\u201dmonitor processes and outcome measures by which the plans participating in the Whole Child Model program shall be monitored and evaluated.\u201dMuch of the research on family engagement has focused on the roles of family engagement in the care of an individual child program director, and highly experienced public health policy analysts and evaluators.The Workgroup was actively engaged in all phases of the project, from design to product development. Members assisted with the synthesis of the literature review, the development of the key informant interview guide, analysis of the findings from the literature review and interviews, identification of the key criteria, and development of the framework for assessing family engagement at the systems level to ensure that family engagement is equitable, authentic, and meaningful.We conducted a literature review, drawing from a variety of sources, within and without the maternal and child health field. This effort included peer-reviewed articles and grey literature reports that highlighted approaches to patient, family, and community engagement and provided a picture of a vibrant and increasingly evidence-based field of study. Using the query \u201ccooperative behavior\u201c[mesh] AND \u201ccommunity participation\u201c[majr] AND (hasabstract[text] AND \u201chumans\u201c[MeSH Terms] AND English[lang]) in Pubmed, we identified 770 articles. The Expert Workgroup recommended an additional 21 articles. We identified an additional 7 articles linked to other articles in the review, for a total of 798, published between 2001 and 2021.5Articles and reports that mentioned family engagement but did not describe how families were supported in such engagement were excluded, as our goal was to identify \u201chow to\u201d ensure that family engagement in systems-level initiatives is meaningful.Using the framework described by Carman et al. , two memThe article demonstrated and/or assessed patient, family, or community engagement at the systems level.Community members participated as members of a core project team.Engagement efforts included training Hawley, or otherCommunity members participated as oversight/advisory council members, key informants, or in other activities that allowed for an iterative dialog between professionals and families.Community members co-developed and analyzed surveys or facilitated and interpreted focus group findings.The project team reviewed the resulting 33 articles, categorized the activities in which families were engaged, and identified the supports the families received to ensure their engagement was meaningful.all children. The key informants, 10 family advocates who represented the diversity of the country, and 9 professionals, were recommended by the Family/Professional Expert Workgroup and by a national network of family-led organizations.To complement the literature review, the project team interviewed 19 key informants to learn about the types of system-level activities families were engaging in as well as the supports that helped families be effective contributors. Interviews also explored what changed because families were engaged and the factors that support meaningful family engagement in policy, practice, and other systems-level activities that impact health services for The family advocates included staff from various family-led and community-based organizations, disability advocates, and members of hospital and other advisory groups. The professionals included public health academics, public and private payers, Title V agencies, regional genetics network personnel, and pediatricians. The interview guide, informed by the findings from the literature review, was the same for both families and professionals.Using NVivo software, research partners at Social and Behavioral Health, School of Community Health Sciences, University of Nevada, Reno performed an analysis of the interview responses.The project team identified the following themes in the 33 articles selected for the literature review:Families were engaged primarily as advisory committee members or as invited attendees at other meetings and selected because they had prior experience or leadership training, mentoring and coaching, peer support, an openness to working in a collaborative setting, and willingness to listen and learn. In some cases, professionals who were also parents of children with special health care needs were asked to assume a dual role as both a family participant and professional.A variety of articles indicated that some \u201csystems of care\u201d understood the importance of ensuring that family partners were representative of the race, ethnicity, culture, and socio-economic status of the families and populations they serve \u00a0that child- and family-serving organizations can use to plan,\u00a0assess, and improve family engagement in systems-level activities over time.Deputy Director, Health Promotion, Los Angeles County Department of Public Health.Deborah Allen, Ph.D. Public Health Research Assistant, University of Michigan; sibling of sister with CP, DD.Amal Alsamawi, Ph.D. PhD, MPH, MBA Director of the CAHMI Professor, Bloomberg School of Public Health Johns Hopkins University.Christina Bethell,* Executive Director, Oklahoma Family Network, F2F in OK; Family Leader.Joni Bruce, Yale School of Public Health.Paul Cleary,* PhD, Executive Director, Support for Families of Children with Disabilities, CA; Family Leader.Juno Duenas, Vice President of Advocacy, Amerigroup, Wellpoint, Inc., VA; sibling of an adult sister with chronic mental health issues and the grandparent of a young boy with a heart condition.Merrill Friedman, MCH Regional Consultant.Carolyn Gleason, President and Chief Executive Officer, Institute for Patient- and Family-Centered Care.Bev Johnson, SVP and Chief Medical Officer Fallon Health; mother of a young adult son with autism and developmental delays.Carolyn S. Langer, MD, MPH, JD, Assistant Professor, Social and Behavioral Health, School of Community Health Sciences, University of Nevada, Reno.Julie Lucero, Associate Professor, Massachusetts General Hospital.Karen Kuhlthau, Ph.D., Professor, Johns Hopkins School of Public Health.Anne Riley, Ph.D., Acting Director, Division of Children with Special Health Care Needs, MCHB.Joan Scott, Assistant Professor, Drexel University, PA; Director of the Pennsylvania Medical Home Program (EPIC IC), Medical Director of Special Programs at St. Christopher\u2019s Hospital for Children.Renee Turchi, MD,"} +{"text": "A wave of new technologies has created opportunities for the cost-effective generation of high-throughput profiles of biological systems, foreshadowing a \"data-driven science\" era. The large variety of data available from biological research is also a rich resource that can be used for innovative endeavors. However, we are facing considerable challenges in big data deposition, integration, and translation due to the complexity of biological data and its production at unprecedented exponential rates. To address these problems, in 2020, the Korean government officially announced a national strategy to collect and manage the biological data produced through national R&D fund allocations and provide the collected data to researchers. To this end, the Korea Bioinformation Center (KOBIC) developed a new biological data repository, the Korea BioData Station (K-BDS), for sharing data from individual researchers and research programs to create a data-driven biological study environment. The K-BDS is dedicated to providing free open access to a suite of featured data resources in support of worldwide activities in both academia and industry. Biological data refer to information derived from living organisms and their products. They are generated from various research areas, including genomics, proteomics, metabolomics, and microarray-based gene expression profiling. In the not-so-distant past, data generation was a bottleneck; now, the rapid technological advancements and the price reductions of new technologies used for biological data production have led to the exponential growth of biological data, the most significant current trend observed in the life sciences . In thisBiological data possess unique characteristics that make biological data management a particularly challenging problem. First, massive amounts and multiple types of 'big' data, such as sequence data, protein structures, and bioimages, are generated from a variety of biological studies. Thus, they are highly complex when compared to other forms of data. Second, since complex biological phenomena involve many aspects, they cannot be fully explained using a single data type. For this reason, the integrated analysis of different data types has attracted increased research attention. However, there are difficulties due to the challenges of heterogeneous data and the implicitly noisy nature of biological data.Many biological databases, such as GenBank and ProtThese global data centers have been committed to preserving and providing access to complete public-domain nucleotide sequences and associated metadata, enabling discoveries in biomedicine, biodiversity, and biological sciences. However, these data centers signaled the possible limiting of the free-of-charge dissemination of data due to the increasing cost of data management. Moreover, considerable challenges are being faced in big data deposition, integration, and translation due to the complexity of biological data and its production at unprecedented exponential rates.Korea is actively producing big data due to large-scale government R&D investments in the biological field. Furthermore, the Korean government officially announced a national strategy for sharing the biological data created using national R&D funds. However, due to the lack of a centralized data center to store such a large amount of data, the reality is that individual researchers own most of the data produced in their biological R&D projects. Thus, the data held by individuals will likely be lost after the completion of the project over time.https://www.kbds.re.kr.To address these problems, the Korea Bioinformation Center (KOBIC) developed a new data repository, the Korea BioData Station (K-BDS), for the last two years and launched it this year. The K-BDS aims to archive biological data through the allocation of national R&D funds and offers unrestricted access to all its publicly available data to the worldwide scientific communities. The purpose of this article is to provide an overview of the recent development of the K-BDS, which is publicly available at The K-BDS is a public repository that manages the biological research data produced by all Korean government-funded research projects, including medical science, health, agriculture, fishery, environmental biology, and many others . The maii.e., metadata and files, where the former can be further assorted into BioProject, BioSample, and experiment, and the latter contains raw data. BioProject, bearing an accession number prefixed with PRJK (where the K stands for Korea from now on), provides an overall description of an individual research initiative, including the submitter, basic project description, funding information, and publication(s) if available. BioSample, possessing an accession number prefixed with SAMK, contains a description of the biological material used in the experiments, including organism, sample types, and attributes. The experiment provides a detailed description of treatments for a specific sample, including library methods and experimental intentions. Raw data are a list of the data file(s) related to a specific experiment.All submitted data are roughly organized into two parts, The biological data in K-BDS are classified into five major groups according to data types. The data types are genomics , proteomics, metabolomics, chemical , and bioimaging . The K-BDS also has the other data type for any other data except the five groups.The accession number is a unique identifier assigned to every single submitted record into the K-BDS. The format of the accession number is [alphabetical prefix][series of digits] . Each acad hoc, and their format depends on how the individual researchers organize various information. Such data are usually organized into a tabular spreadsheet form, similar to the supplementary information data that accompanies journal publications. For such unstructured data, the submission forms of the existing generalist repositories, such as BioStudies [The K-BDS is a repository for all types of data for biological research. Therefore, data submission forms were developed for each of the data types. Depending on the data type, different strategies were adopted to establish a data submission form that could make the inputted information sufficient and valuable. The formats of the data types are divided into three cases. First, for some data types, there is either an official standard file format or a de facto standard file format . Such data types also usually have minimum information guidelines on how to prepare their metadata as well as established data archives specific to them, an example for functional genomics data is the Minimum Information About a Microarray Experiment (MIAME) guideline and the oStudies and FigsoStudies , were reIn developing the submission forms for all three cases, as mentioned above, for the various data types, experts from diverse areas in the Korean biology community participated in maximizing the submission forms' information content and scrutinizing them. A total of approximately 130 experts participated in the development of the data submission forms from 2020 to 2022. The submission forms are grouped into five major categories of data types and some other categories.The data submitters upload and submit any biological research data using the K-BDS submission system . Users mThe BioProject is a collection of biological data related to a research effort. A BioProject submission is required for biological data submissions regardless of the data type to be submitted. The BioProject stores information on authors and grants, project information, related publications and patent information if available. A BioProject record provides users with a single place to find links to the diverse data types generated for that project.BioSample stores the descriptions and metadata about the biological samples used in the studies. BioSample uses a batch submission process and accepts diverse data types, including human, plant, animal, microbe, virus, and metagenome. BioSample examples include a cell line, a primary tissue biopsy, and an individual organism or an environmental isolate. For molecular biology fields such as genomics, proteomics, and metabolomics, a BioSample submission is required before submitting the experiments and raw data. BioSample also provides reciprocal links to BioProject, facilitating sample searches.The K-BDS has different data submission processes according to the biological data type after inputting data into BioProject and BioSample. The user first selects the data type to be registered, then checks the standard data form to prepare the data to be input, and then inputs the data to the K-BDS registration site. Users can download the user submission manuals for each data type from the K-BDS website. In addition, the K-BDS homepage provides videos for registrants and users so that they can be used more efficiently.When researchers submit their data, temporal numbers are immediately assigned. These numbers are only used to manage the data and are only revealed to the owners. Then, the accession number is assigned and disclosed to other researchers after completing the quality control process.As of December 2022, the K-BDS contains 289 biological projects, 45,108 samples, and 1,484,822 data records . For NGSWe developed a fast file transfer tool called GBox, for uploading massive data quantities such as entire genomes and exomes to the K-BDS server from the user's local computer and downloading the selected files to the local computer. The GBox client program can be downloaded from the K-BDS website and installed on the user's computer. The GBox transfer platform provides users with the secure high-speed movement of their data, supporting a wide range of servers, desktops, and Linux operating systems. Using the K-BDS, users can simultaneously upload unlimited numbers of files. GBox has a file transfer speed of approximately ten times that of the standard FTP and HTTP protocols.The most crucial thing in database development is the quality of the data and its metadata. Data quality indicates a dataset's reliability across key dimensions such as completeness, consistency, and accuracy. Thus, the data quality management system was built to check all the submitted data before issuing an accession number.The data quality control process in the K-BDS has two steps, the automatic step, where records are automatically checked by software without a manual review, and the expert step on selected records. A significant task in the automatic step is to inspect records according to the rules configured in the standard registration form. The submission process does not proceed if the input data are invalid.For the expert step, domain experts run a comprehensive set of software, search the supporting information from various databases, manually review the results, and interpret the evidence level. The K-BDS has five domain expert groups for genomes, proteomes, metabolomes, chemicals, and bioimages. KOBIC is in charge of genome data, and extramural external expert groups in Korea are designated to perform more professional quality control jobs for the other four data types.The K-BDS provides user-friendly web interfaces for data queries and browsing. A simple text keyword search allows users to find relevant data. Users can search the data of interest by specifying a given ID of the BioProject, BioSample, or dataset. Moreover, the K-BDS allows users to conduct an advanced search by inputting species names, titles, diseases, and tissue, for example. The K-BDS also allows users to browse all publicly available BioProjects, BioSamples, and datasets.https://www.egovframe.go.kr/eng/main.do), Java Server Pages (a Java programming framework for constructing dynamic web pages), MyBatis (a persistence framework for the database connection and operation), MySQL , and Elasticsearch . To provide a stable web service, K-BDS is hosted on five servers (CentOS 7), an HTTP server for static content, a tomcat server for dynamic range, a MySQL server for database management, an Elasticsearch server for data retrieval, and an FTP server for data download.The K-BDS is implemented based on multiple frameworks, the e-Government Standard Framework when data are stored in the K-BDS system, they are indexed and fully searchable in near real-time; (2) the support of high-speed retrieval for structured and unstructured data using a big data platform capable of parallel distributed processing; (3) the support of the flexible extension of diverse data types using schemeless NoSQL; (4) the support of diverse programmatic access methods through supplying RESTful APIs; (5) visualization of real-time statistics from access logs using elastic search and Kibana tools.The hardware design goal of the K-BDS system is to provide large-capacity storage and high-performance computing based on a big data analysis platform for large-scale data. The specifications of the K-BDS infrastructure are as follows: 1,260 CPU cores, 4 NVIDIA A100 80 GB (SXM4) GPUs, 24 TB memory, Mellanox InfiniBand (200 Gb/s), 18.6 PB storage with an additional 18.6 PB backup.The storage system consists of a DDN ES7990X and a Lustre parallel file system for large-scale data storage and high-speed, low-latency, high-bandwidth data transfer are supported through InfiniBand HDR. The computing servers for extensive data analysis are composed of 1,260 CPU cores, NVIDIA A100 GPUs, 12 TB memory with a Sun Grid engine for distributed processing, and the docker to support various heterogeneous analysis programs. The K-BDS platform uses the PIOLINK PAS-K3200 L4 Switch to perform redundancy and load balancing with the slightest connection algorithm method, WEB (Apache 2.4.53) and WAS (Apache-Tomcat-9.0.68).MySQL 5.7.26 is used for the database management system. The data backup system consists of primary storage snapshots, backup storage disks and remote backup disks (in the Ochang data centers). To prepare against disasters and disabilities, a disaster recovery system operates in the Ochang data centers. Moreover, K-BDS provides data analysis services with Bio-Express , cloud-bBio-Express is a scalable, cost-effective, and publicly available web service for large-scale genomic data analysis. Bio-Express supports the reliable and highly scalable execution of sequencing analysis workflows in a fully automated manner. The Bio-Express provides a user-friendly interface to all genomic scientists to try to derive accurate results from NGS platform dataTo summarize, the K-BDS is a data repository for archiving raw biological data and providing free access to various database resources supporting research activities. Designed for compatibility, the data structure of the K-BDS adopts international data standards and formats.The K-BDS has three major advantages. First, the K-BDS is a public repository for the biological research data produced by all Korean government-funded research projects. Therefore, various types of data can be searched on the K-BDS websites. For example, genome, proteome, and image information derived from the same sample can be simultaneously searched and downloaded. Second, the K-BDS provides a web-based user interface for data submission that is simple and easy to use. We also developed a fast file transfer tool, GBox, for uploading massive biological datasets to the K-BDS server from the user's local computer. Lastly, the K-BDS provides a cloud-based analysis function, Bio-Express, for data registered in the K-BDS. Bio-Express enables the data to analyze without downloading the data into the local computer.The K-BDS is the new data repository that officially started in December 2022. Therefore, the functions and d services of the K-BDS are not the same as those of world-class data centers such as NCBI, EBI, and DDBJ, which have been in operation for more than 30 years. However, due to the active investment of the Korean government and the development of data-based science, the level of KBDS will rise to the level of a world-class data center in a short time.The ultimate goal of the K-BDS is to establish and promote a centralized archival practice in Korea and support research activities in both academia and industry throughout the world. Ongoing efforts include optimization and automation of the data submission, curation and analysis procedures, an infrastructure upgrade for big data storage and transfer and development of new tools and pipelines to support worldwide biological research.Sharing data is more useful when others can easily find, access, interpret, and reuse the data. To maximize the benefit of sharing data, the K-BDS will continue to expand and offer a series of data resources and services to benefit a wide range of research initiatives in the life and health sciences."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "In the context of synthetic biology, light and other electromagnetic radiation provide a powerful tool to drive and control parts, devices, systems and processes with high target specificity and switching efficiency. Thanks to its peculiar features in terms of wavelength and energy, light allows spatio-temporal control of molecular and supramolecular systems that ultimately lead to (bio)chemical and (bio)logical behavior\u2014such as optogenetic control and energy fueling.in vitro or in situ/in vivo, that convincingly show the versatility and the power of such approaches. Examples involve both visible and near-infrared light in bacteria, yeast, mammalian and plant cells. The light control of gene expression has been used for triggering processes such as signaling, recombination, initiation of translation, production of chemicals and peptides, apoptosis, intracellular transport, and cell differentiation. Additional cases refer to protein localization distribution, protein degradation, protein homo- and hetero-dimerization, alteration of a protein\u2019s metal binding behaviour, and also protein coacervation. Complex processes including biofilm formation, cell differentiation and morphogenesis have been considered.In the recent years, a large number of studies have been devoted to the design and construction of molecular systems, either All these achievements excitingly impact current synthetic biology research because of the intrinsic versatility and power of light, which makes it easier to drive and control a large number and variety of processes.Fiat Lux! Light-Driven and Light-Controlled Synthetic Biological Parts, Devices, Systems and Processes\u201d, which was focused on the above-mentioned approaches, collects seven articles that deals with the subject of light-driven and light-controlled synthetic systems and processes, in particular optogenetics, but also related approaches. The resulting picture provides a cross section of the state of the art in the field, and can inspire and guide further investigations.The Research Topic \u201cFiat Lux! Light-Driven and Light-Controlled Synthetic Biological Parts, Devices, Systems and Processes\u201d counts seven articles. The five reviews and the perspective paper can be advantageously utilized as a Research Topic of discussions about relevant issues in light-controlled biodevices: from their functioning to the investigation platform, from their design to applications.The Research Topic \u201cOhlendorf and M\u00f6glich review the fundamentals, the advances, and the perspectives of optogenetically regulated gene expression in bacteria. By highlighting three fundamental strategies, namely, light-sensitive two-component systems, oligomerization, and second-messenger signaling, the authors highlight relevant application areas of optogenetic control. One is the enhanced yields that can be achieved in microbial production processes. Next, the control of the secretion of compounds that grant health benefits to the animal host by light-responsive bacteria which reside within the bodies of animals. Third, optogenetics can lead to the synthesis of precisely structured, novel biomaterials.Two comprehensive reviews must be mentioned firstly. Baumschlager, on the other hand, presented a review that discusses important aspects for engineering of light-controllable proteins through selected examples. The focus is on non-neuronal optogenetics, chromophore availability, general strategies for creating light-controllable functions, modification of the photosensitive domains and their fusion to effector domains, as well as tuning concepts for opto-proteins.Dwijayanti et al., focused their review on the progress made on systems with multiple photoreceptors, each sensing its dedicated wavelength. The combination of multiple photoreceptors allows a coordination of cellular responses. Recent works and challenges on multiplexed optogenetic circuits in natural and engineered systems are discussed.The next two articles deal, instead, with more specific research topics. In particular, Zhanget al., instead, is a perspective article on light switchable two-component protein dimerization systems. The study provides categories for mechanisms and design approaches of these dimerization systems, which have been recently progressed by the discovery of photoreceptor-based interaction systems, by the engineering of light-actuatable binder proteins, and by the development of photoactivatable compounds as dimerization inducers.The contribution written by Kumar and Khammash, who reviewed the evolution of light-induction hardware-software platforms from simple illumination set-ups to sophisticated microscopy, microtiter plate and bioreactor designs, and discuss their respective advantages and disadvantages. Moreover, experimental approaches such as treatments of different cell types and culture volumes, with induction capabilities ranging from single cell stimulation to entire cell culture illumination, automated measurement and stimulation schemes on these platforms.The methodologies and experimental platform for studying optogenetic controls have been thoroughly discussed by M\u00e5nsson et al. illustrated, in another review, mechanisms and applications of \u201cOptoGels\u201d, i.e., hydrogels with light-programmable properties endowed by photoswitchable proteins (\u201coptoproteins\u201d) found in nature. Thanks to conjugation chemistry OptoGels with a combinatorially large design space (still not well explored) can be designed, resulting in a rich variety of tunable material properties. The potential future applications of OptoGels range from mechanobiology to 3D cell and organoid engineering, as well as programmable cell eluting materials.On the other hand, Hilgers et al. Their investigation focuses on photocaged inducer molecules as well-established optochemical tools for regulating bacteria gene expression by light. The study is based on the photoactivation of gene expression in Rhodobacter capsulatus by using different cIPTG variants, under phototrophic and non-phototrophic cultivation conditions. The authors have identified a promising compound, 6-nitropiperonyl-(NP)-cIPTG. The optochemical approach was then successfully applied to the induction of carotenoid biosynthesis.An original research article (the only one in the Research Topic) has been presented by Albanese et al. have reported about the latest strategies for the assembly of energetically autonomous \u201cbottom-up\u201d artificial cells. The authors refer to the branch of synthetic biology devoted to the construction of cell-like systems able to mimic fundamental aspects of living cells, yet being minimally complex. The review focuses on the exploitable (and exploited) bio-inspired mechanisms of light transduction aimed at supporting internalized metabolic pathways in artificial cells. The concepts behind the construction of mono- and multi-compartment artificial cells capable of light-driven proton gradient and ATP production are systematically presented and discussed in the context of next-generation artificial cells.Finally, The Research Topic of the above-mentioned papers is of great interest for synthetic biologists who intend to learn about the state-of-the-art in using light to control biological processes. The articles collectively offer a wide perspective on a variety of approaches and investigations carried in the past years on optogenetic control and related systems and on the bottom-up construction of cell-like systems that exploit light as ultimate energy source."} +{"text": "ADHD is one of the most frequently diagnosed neurodevelopmental disorders and affects the daily functioning of families raising children with this condition. Among the symptoms typical for ADHD, low effectiveness of executive functions can determine the quality of family life.This study aimed to specify whether family communication and satisfaction as reported by a parent are predictors of a child\u2019s executive functioning quality and whether ADHD severity lies on the pathway between the two. Moreover, the child\u2019s sex effect was checked.The study included 200 Polish participants (nGirls = 56) from the NeuroSmog project aged 10-13 diagnosed with ADHD according to the ICD-11. Stanford-Binet 5 Intelligence Scale, PU1 Cognitive Diagnosis Battery, Conners 3 ADHD Diagnosis Questionnaire, and the FACES IV Questionnaire were used to derive needed information. Structural equation modelling (SEM) was applied to test the hypotheses.The quality of family communication and satisfaction did not predict the child\u2019s executive functioning of ADHD children and ADHD severity did not play a mediating role. No differences by sex were observed. We only found a significant effect between IQ and executive functioning level in the general sample and in girls .These results contrast with previous studies from other cultural contexts that have shown the existence of the hypothesized interrelations. Further research should confirm or refute these observations.W. Walenista Grant / Research support from: the TEAM-NET programme of the Foundation for Polish Science, co-financed from EU resources, obtained from the European Regional Development Fund under the Smart Growth Operational Programme, B. Izydorczyk Grant / Research support from: the TEAM-NET programme of the Foundation for Polish Science, co-financed from EU resources, obtained from the European Regional Development Fund under the Smart Growth Operational Programme, K. Sitnik-Warchulska Grant / Research support from: the TEAM-NET programme of the Foundation for Polish Science, co-financed from EU resources, obtained from the European Regional Development Fund under the Smart Growth Operational Programme, I. Markevych Grant / Research support from: the TEAM-NET programme of the Foundation for Polish Science, co-financed from EU resources, obtained from the European Regional Development Fund under the Smart Growth Operational Programme, C. Baumbach Grant / Research support from: the TEAM-NET programme of the Foundation for Polish Science, co-financed from EU resources, obtained from the European Regional Development Fund under the Smart Growth Operational Programme, Y. Mysak Grant / Research support from: the TEAM-NET programme of the Foundation for Polish Science, co-financed from EU resources, obtained from the European Regional Development Fund under the Smart Growth Operational Programme, M. Szwed Grant / Research support from: the TEAM-NET programme of the Foundation for Polish Science, co-financed from EU resources, obtained from the European Regional Development Fund under the Smart Growth Operational Programme, M. Lipowska Grant / Research support from: the TEAM-NET programme of the Foundation for Polish Science, co-financed from EU resources, obtained from the European Regional Development Fund under the Smart Growth Operational Programme."} +{"text": "Lancet. (2021) 397:10277. doi: 10.1016/S0140-6736(21)00458-X.Following concerns regarding the originality of the article, an investigation was conducted in accordance with Frontiers' policies. It was found that the complaint was valid and that the article should be retracted, because of an unacceptable level of similarity to an article published by Meghji J, Mortimer K, Agusti A, Allwood BW, Asher I, Bateman ED, et al. Improving lung health in low-income and middle-income countries: from challenges to solutions. This retraction was approved by the Chief Editors of Frontiers in Public Health and the Editor-in-Chief of Frontiers. The authors did not agree to this retraction."} +{"text": "Correction: Aging Clinical and Experimental Research (2020) 32:2141\u20132158 10.1007/s40520-020-01677-yIn the Acknowledgements section of this article the grant number relating to National Institutes of Health given was incorrectly given asThe authors would like to thank Benjamin D Keane for his assistance in preparing the manuscript. The authors would also like to acknowledge generous support from the Carinato Charitable Foundation, Mark and Ingeborg Holliday, Kristin Hudson and Rob Goldman, and Ms. Susan Brice and Mr. Jordi Esteve.and should have beenThe authors would like to thank Benjamin D Keane for his assistance in preparing the manuscript. The authors would also like to acknowledge generous support from the Carinato Charitable Foundation, Mark and Ingeborg Holliday, Kristin Hudson and Rob Goldman, and Ms. Susan Brice and Mr. Jordi Esteve. In addition, the authors would also like to acknowledge funding from the National Institutes of Health (NIH), through grant #R01CA225002.The original article has been corrected."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Communications Biology 10.1038/s42003-023-05054-z, published online 27 June 2023.Correction to: In this article the funding from the \u2018Deutsche Forschungsgemeinschaft , the State of Thuringia (TMWWDG), and the Free State of Thuringia , and support by the integration into the Leibniz Center for Photonics in Infection Research \u2019 was omitted. The original article has been corrected."} +{"text": "Morningness (morning-eveningness preference or chronotypes) and personality can be both associated with well-being, but few studies have directly compared these two constructs as correlates of well-being. Thus, the first purpose of this study was to test the effects of interactions between stable personality traits (temperaments) and morningness on well-being. Furthermore, personality factors are often composed of both stable biological factors (temperament) and socio-cultural factors (character), and little is known about personality interplay of temperament and character factors with respect to morningness and well-being. The second purpose of this study was therefore to examine the sequential mediating effects of temperament and character factors on the relationship between morningness and well-being.The Composite Scale of Morningness, the Korean version of the Temperament and Character Inventory-Revised Short Version (TCI-RS), and the Satisfaction with Life Scale were used to measure morningness, personality dimensions, and well-being, respectively, in 287 Korean university students. Moderating and sequentially mediating effects of temperament and character traits were determined using Hayes\u2019 PROCESS macro in SPSS after controlling for sex and age.First, novelty-seeking (NS) and persistence (PS) temperaments have demonstrated the moderating effect in the association between morningness and well-being. The positive effects of morningness on life satisfaction increased with lower NS and PS, respectively. However, other temperaments such as harm avoidance (HA) and reward dependence (RD) have not shown the moderation in the relationship between morningness on well-being. Second, HA temperament and self-directedness (SD) character sequentially mediated the relationship between morningness and well-being. The combination of low scores of HA and high scores of SD have shown the positive effect on the relationship between morningness and well-being.This study demonstrated that both the interactions between temperaments and morningness, and combination of specific TCI-RS temperament and character traits play important roles in influencing the association between morningness and well-being. The significance of the mature SD character and its implications for well-being are discussed with limitation of the present study. Morningness, or chronotype, exists on a continuum that manifest as individual variation in circadian rhythm . The morMany studies have described the relationship between morningness and various individual differences, one of which is subjective well-being . MorningExtensive research have been conducted on the relationships between morningness and personality factors using, fUnlike the well-established linear associations between the FFM personality traits and well-being, the relationship between TCI temperament factors and well-being is somewhat inconsistent: NS and PS have shown mixed results, while HA is primarily negatively correlated and reward dependence (RD) is not correlated with well-being .via interactions between the biological being and the environment. It also focuses on the possibility of complex and dynamic systems that exert an effect in the within-person development of personality and well-being . Character, on the other hand, is what people consciously make of themselves intentionally. Accordingly, character regulates the way our temperament makes us react to the internal and external environment , cooperativeness (CO), and/or self-transcendence (ST) items of the TCI are protective against psychopathology and promote mental health .e.g., low HA) and high mature-character dimensions is associated with increased well-being accounts for a significant amount of shared variance between the independent and the dependent variables .Within the framework of the TCI, the combination of HA and SD scores often plays an important role in well-being and psychopathology . That isll-being . For exall-being . TherefoThis study aimed to explain the key roles of the temperament and character dimensions of TCI in biopsychosocial functioning associated with the relationship between morningness and well-being.The Composite Scale of Morningness (CSM), TCI-Revised-Short Version (TCI-RS), and Satisfaction With Life Scale (SWLS) were used to measure morningness-eveningness, temperament and character, and life satisfaction, respectively, in a sample of 287 university students in Busan, South Korea. Out of 311 students, 287 volunteered to participate and completed our questionnaire. Participants were also asked about their age and sex. This study was approved by the Institutional Ethics Board of Kyungsung University, South Korea and all participants provided written informed consent.The CSM is commoThe TCI-RS consists of two interrelated domains of temperament and character, and is originally developed by The temperament dimensions comprise NS , HA , RD , and PS . The character dimensions are SD , CO , and ST .The TCI-RS Korean version is a 140-item self-report questionnaire which requires individuals to score each item using a 5-point Likert scale. The internal consistencies of NS, HA, RD, PS, SD, CO, and ST were reported to be 0.83, 0.86, 0.81, 0.82, 0.87, 0.76, and 0.90, respectively . The intThe SWLS is a five-item self-report questionnaire which measures an individual\u2019s subjective life satisfaction or well-being on a 7-point Likert scale . The SWLSignificant differences between male and female participants in age, morningness, well-being, and TCI-RS personality dimensions were examined using t-tests. Pairwise correlations between morningness, well-being, temperament and character items were determined using Pearson\u2019s correlation, and correlation coefficients were provided.Two hypotheses were tested: the first (H1) was that NS and PS temperaments would moderate the association between morningness and well-being; the second (H2) was that HA and SD would sequentially mediate the relationship between morningness and well-being. For both hypotheses, moderation and mediation analyses were conducted using Hayes\u2019 PROCESS macro , after controlling for sex and age.Additionally, a floodlight analysis was conducted using thThe sequential mediation analysis was performed using ordinary least squares regression to estimate direct and indirect effects. Effect size estimates of indirect associations were drawn using bootstrap confidence intervals.p values <0.05 was considered statistically significant. IBM SPSS Statistics 20.0 was used for all statistical analyses.The data are presented as means and standard deviations, and t = 2.975, p < 0.01), well-being , NS , HA , and ST but not in CSM , RD , PS , SD , or CO between men and women. The higher male age is a consequence of men in South Korea often having to complete their mandatory military duty in their lower school years. Considering this finding and the significant sex-related differences identified, we performed all analyses with age and sex as covariates.The demographic features of the participants are presented in p < 0.05), SWLS , NS , HA , PS , SD , or CO . The SWLS total score (higher score represents greater life satisfaction) was significantly correlated with HA , PS , SD , CO , or ST .Correlations between the CSM, SWLS, and TCI-RS response data were determined using Pearson\u2019s correlation analysis. The CSM total score (a higher score represents a greater tendency toward morningness) was significantly correlated with age , although the effect of morningness and the interaction term were significant . The results showed that the positive effect of morningness on well-being increased with lower levels of NS in SPSS. The first regression model, including morningness, NS temperament, and an interaction term after controlling for age and sex, proved significant, F = 7.507, ls of NS . The flop < 0.001, and explained 21.7% of the variance in well-being. The effects of PS and morningness were both significant along with that of the interaction term . The results showed that the positive effect of morningness on well-being increased with lower levels of PS = 15.537, ls of PS . The floOther regression models including HA and RD temperaments and morningness were conducted, but no interactions between temperament and morningness were observed. Therefore, the following procedures were not tested.\u03b2 = 0.2433, p < 0.001). However, the effect of morningness on life satisfaction decreased after the inclusion of HA and SD in the direct-effect model. This reduction in the magnitude of the effect of morningness without a decrease in the significance level indicates partial mediation of well-being outcomes. To confirm the partial mediation effect of HA and SD, the indirect effects were scrutinized.To test H2, sequential mediation analysis was conducted using Hayes\u2019 PROCESS macro (Model 6) in SPSS. The full process model, showing all path coefficients, is presented in \u03b2 = 0.0201, \u03b2 = 0.0265; respectively). However, the indirect effect of morningness on life satisfaction through the sequential mediation of HA and SD was significant (\u03b2 = 0.0753). The indirect effect (0.069) to total effect (0.1277) ratio in the mediation was 0.54, which showed the meaningful mediation effect. This finding provides support for the sequential mediation model in the context of personality and well-being. HA and SD failed to independently mediate the relationship between morningness and life satisfaction, while HA and SD partially mediated this relationship in sequence. Finally, the sequential mediation effects of HA and the other character dimensions of CO and ST were not supported.The data in Many studies have investigated the relationship between morningness and personality, and between personality and well-being. However, there is a paucity of research examining the direct relationship between morningness and personality, and their impact on well-being. We used the TCI-RS to study these features, which we consider is the most appropriate personality instrument for this purpose. The TCI-RS and the well-being construct share the fundamental concepts of temperament and character; in other words, well-being is influenced by innate and biological temperament factors and by mature and socio-cultural character features.Through the use of the TCI-RS, we found the moderating effects of both NS and PS on life satisfaction according to chronotype, while HA and RD did not. The interaction effect of morningness and NS was significant when the Johnson-Neyman point was lower than 9.695 but not significant when the Johnson-Neyman point was higher than 9.695. This might mean that the effective regulation of the relationship between morningness and well-being only occurs when NS is low. Approximately 84% of the participants experienced this effect when NS was below 9.695. In the lower-NS group, morningness was associated with increased well-being, but there was no difference in well-being in the higher-NS group, regardless of chronotype.Similarly, the interaction effect of morningness and PS was only significant when the Johnson-Neyman point was lower than 5.821. This means that the relationship between morningness and well-being was moderated only when PS was low. The significance rate of this effect was approximately 72% when PS was below 5.821. In other words, morningness in lower-PS individuals was associated with greater well-being, but there was no difference in well-being in higher-PS participants, regardless of morningness or eveningness.In particular, the finding of moderation by NS may explain the mixed results of previous studies . These sPS also had a moderating role in the relationship between morningness and well-being, and this factor also demonstrated a simultaneous positive correlation with morningness and well-being. Similar to the NS trait, excessive PS may not have a positive effect on well-being. The high PS score tendency showed the advantage and disadvantage effects in emotional regulation, an important subfactor of well-being . A persoAccording to a definition proposed by Cloninger, temperament traits such as NS and PS have a nonlinear association with character and well-being . The levSecond, considering the importance of character in controlling temperament and influencing well-being, the sequential mediating effect of temperament and character was examined in the present study. The serial mediating effect of HA and SD was significant; however, NS, RD, and PS temperament traits did not show such an effect with SD. In other words, the partial mediating effect of personality on morningness and well-being was supported: morningness itself not only affects well-being, but the combination of personality variables such as HA temperament and SD character dimension exerts additional influence.SD is an important prerequisite for character development that emphasizes protective factors, considering that HA is negatively correlated with well-being. This is consistent with previous studies . IndividNumerous studies have shown that SD is directly and linearly related to well-being. The character domain of TCI is a measure of psychological maturity, an apparent protective factor for psychopathology, and is related to biopsychological well-being . The impCollege students usually have a circadian rhythm with an evening preference and are, therefore, presumed to experience reduced well-being. However, the results of the present study imply that the combination of the HA temperament and SD character plays a complementary role in the development of well-being, even among evening-type people. Therefore, future therapeutic interventions could focus on the promotion of SD, given the supposed stable and unchangeable nature of temperament traits. Well-being could be enhanced through the development of SD since it is a character variable that controls temperament .There are several limitations to generalizing the findings of this study. First, although this study was performed on Korean university students with demographic features, the hypotheses should be examined in a larger sample size with a balanced age and sex distribution.Second, our study was cross-sectional, which restricts any interpretation of causality. A well-designed, longitudinal study is required to test the temporal relationship between morningness and personality and its effects on well-being. Even though there is one longitudinal study regarding the effect of morningness on subjective well-being , it is se.g., positive and negative affect) and/or non-affective components . Therefore, life satisfaction is just a part of well-being, even though we used both terms interchangeably in current study, as have other studies (Third, well-being consists of affective ( studies . FurtherFourth, there is a report emphasizing the importance of circadian amplitude when considering the relationship between well-being and morningness-eveningness in college students . MeasuriIn summary, this study explored how the interplay between morningness and temperament influence well-being by examining moderating effect of temperament on the relationship between morningness and well-being. Furthermore, this study provided significant insight into the influence of temperament and character dimensions on the effects of the relationship between morningness and well-being by inves10.7717/peerj.15861/supp-1Supplemental Information 1Click here for additional data file."} +{"text": "Scientific Data 10.1038/s41597-023-02234-8, published online 01 July 2023Correction to: Qiyue Liu, North China Institute is Aerospace Engineering, was incorrectly omitted as an author of this work in the original version. This has been corrected via additions to the author list and author contributions statement in the HTML and pdf versions of the paper."} +{"text": "Studies related to attitudes toward the use of prohibited substances in Turkish athletes are scarce. The World Anti-Doping Agency (WADA) has implemented anti-doping educational policies emphasizing doping-related education in studies conducted among Turkish wrestlers. However, it is still unclear the extent to which the wrestlers comply and adhere to these anti-doping policies. No research has previously examined the effect of anti-doping education on athletes' mindfulness and moral disengagement in doping (MDD). Therefore, the present study has a two-fold objective: first, to examine whether doping-related education (DRE) and the status of being a national athlete (NA) have an effect on athlete mindfulness and MDD. Second, to analyze the relationship between each sub-dimensions of athlete mindfulness: awareness (ASD), judgment (JSD), and refocus (RSD) with MDD.A total of 409 male wrestlers participated in this study. MANOVA analysis showed that NA and DRE alone have no effect on MDD but have a general effect on mindfulness.0.173). When the interaction effect of NA*DRE was examined, significant difference in MDD , ASD , JSD , and RSD were found. MDD has a weak negative relationship with ASD (r = \u22120.126) and RSD (r = \u22120.041) and a weak positive relationship with the JSD sub-dimension (r = 0.140). Those results suggest that being a NA and having received anti-doping education affect moral disengagement in doping and athletes' mindfulness.The highest effect was on the ASD of being an NA (As a conclusion, it is recommended to increase awareness and anti-doping education among national-standard Turkish wrestlers to prevent them from engaging in doping behaviors. Mindfulness is a psychological process that progresses by avoiding judgments and being aware of the individual's experiences in the present . The data collection process took place between 26 and 31 July 2022. The convenience sampling method was used while collecting the data. Participants were asked to fill out the form immediately within a maximum of 30 min. Data were collected by reaching out to universities' wrestling teams and sports clubs. Participants were athletes who had received education on doping at the universities.In this cross-sectional study, the convenience sampling method was conducted on 426 wrestlers who participated in the study . A totalParticipation in this study was entirely voluntary. Any participant had the right not to participate or quit the study at any time after participating. The participant's filling in the questions on the measurement tool means that he gave his consent to participate in the research. It was stated that no one should be under pressure while answering the questions, and that the data obtained from the study will be used for research purposes only. This study was conducted in accordance with the tenets of the Declaration of Helsinki. Ethical approval was obtained from the Ethics Committee of Tekirdag Namik Kemal University.The first part of the questionnaire includes demographic information consisting of doping-related education status and being a national athlete. In the second part, the Mindfulness scale was used, which has been used in previous studies meets the assumption of normality for continuous variables and frequency and percentages for categorical variables. In order to assess the relationship between the variable of interest, Pearson correlation analysis was performed. In the study, the MANOVA test was used to examine the main effect of a national athlete (NA), doping-related education (DRE), and the interaction effect of NAWhen Cronbach's alpha reliability analysis was examined, the result was 0.79 in the awareness sub-dimension (ASD), 0.79 in the judgment sub-dimension (JSD), 0.81 in the refocusing sub-dimension (RSD), and 0.86 in moral disengagement in doping (MDD) level. All statistical analyses were conducted using SPSS software . Statistical significance was set at alpha level < 0.05. American Psychological Association (APA) 6.0 style was used to report statistical differences . According to results, it was determined that the highest effect was on the ASD of being an NA , and conducting this study in different sports will make a significant contribution to the field of sports psychology.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The study was approved by Tekirdag Namik Kemal University Social and Human Sciences Scientific Research and Publication Ethics Committee (dated 23.06.2022 and numbered 173291). The patients/participants provided their written informed consent to participate in this study.EA: conceptualization, methodology, and investigation. HK: software, resources, visualization, and data curation. FY and HK: validation. FY: formal analysis. HK and EA: data curation. EA and MG: writing of original draft preparation. EA, MG, EG-G, MA, and SA-M: writing of review and editing. MA: funding acquisition. All authors contributed to the article and approved the submitted version."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.1As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.3Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.5Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.7Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.16Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.22Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.10In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "In the last decade, the use of electric bikes (e-bikes) has become increasingly common due to rapid advances in technology. This has led to an increased number of publications aimed at investigating individual factors and beliefs that influence a modal shift towards e-bikes. To our knowledge, there is currently no review summarizing the evidence on individual motivations, facilitators, and barriers for e-bike use. This review aims at providing an overview of the current state of the art regarding individual choices and intrinsic factors to change from other modes of transport to e-bikes.Searching for the term e-bike its synonyms and permutations on the databases SCOPUS, PubMed, Web of Science, and PsychINFO resulted in a total of 3,317 hits after removing duplicates. Screening of title/abstract and full-text screening is currently being conducted independently by three researchers using Covidence. Studies reporting on personal decision-making factors and reasons for a modal shift to a private e-bike are considered eligible for inclusion in this study. Studies focusing on public e-bike services are excluded. Studies will be analyzed regarding (a) research quality, (b) factors and reasons for model shift, and (c) information about the investigated populations.Preliminary results show a substantial increase in scientific studies related to e-bikes. After the title and abstract screening, 121 studies were included for full-text screening. Preliminary results allow assumptions about two different aspects: (a) Most studies connected the modal shift to e-bikes to facilitators rather than to barriers. Financial, temporal and social factors, along with enjoyment, health and ease of commuting seem to be among the motivators to change to an e-bike as a means of transportation, (b) the majority of studies focused on middle-aged and older populations.The identified articles and their summary show the increased relevance of e-bikes in current research and public health. Our results may contribute to understanding the subjective reasons why people change their mode of transport to the e-bike. This knowledge may be useful for implementing public health interventions, infrastructure developments, and policies. More research is needed to better understand these factors and their impact."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Correction to: Discover Nano (2023) 18:74https://doi.org/10.1186/s11671-023-03845-1The original article [1] contained an error in the affiliation of author Muhammad Shafiq Shahid:The author is affiliated with the Department of Plant Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, and not with the Department of Plant Pathology, University of Agriculture, Faisalabad 38,040, Pakistan, as previously indicated in the article.The correct affiliation is shown in the 'Author details' of this Correction, as well as in the original article, which has now been updated."} +{"text": "Dear Editor,Orthopox genus, spreads through zoonosis and in habitats rodents, mammals, and primates as an animal reservoir making it more susceptible to transmission and infectiveThe COVID-19 pandemic has imposed a global threat to all the developed and developed countries targeting social, economic, and political platforms influencing the global interconnections in 2020. In the era of ongoing long-term COVID-19 pandemic post-effects worldwide, spontaneous, and unexpected monkeypox outbreaks have attained global consideration. At the global platform, apart from its native origin central and western Africa reported monkeypox as sporadic, exceptionally huge, and massive 3340 confirmatory numbers of monkeypox epidemic were reported on 22 June 2022, at the global level. Monkeypox virus (MPXV) is a member of the In 2022, a total of 3413 confirmed cases with one death reported from 1 January 2022, to 22 June 2022, with 86% of cases from the European region from 50 countries in five WHO regions. Among these 3413 total cases from 3 May 2022, to 22 June 2022, 3340 cases were identified from nonendemic MPXV of 42 Member States in four WHO regionshttps://www.cdc.gov/poxvirus/monkeypox/response/2022/world-map.html). In Ukraine, the first incidence of monkeypox was validated in a laboratory on 15 September 2022 (https://moz.gov.ua/article/news/v-ukraini-zareestrovanij-vipadok-vispi-mavp). The current situation in Ukraine reported no introduction of mass vaccinations as per reduced registered cases of monkeypox as per WHO. However, in July as per global humanitarian help for the diagnosis of MPXV, 5000 PCR kits were delivered to Ukraine and aided in diagnosis regionwide (https://www.manilatimes.net/2022/09/17/news/world/ukraine-records-first-monkeypox-case/1858715). The infection was diagnosed owing to PCR research undertaken by the Regional Centre for Disease Control and Prevention of Ukraine\u2019s Ministry of Health. This case is now being investigated epidemiologically. The patient claimed that he had not gone outside of the country and had no encounters with monkeypox sufferers. Nonetheless, the advent of infectious symptoms and the initial findings of the epidemiological records indicate that the individual was affected in one of the nation\u2019s major cities. The patient was found to have an elevated body temperature and rashes on his body as symptoms of the infection. Doctors in Ukraine follow recognized WHO standards and international methods to diagnose and treat monkeypox. Given that medications are inaccessible to the general population, it is essentially the subject of symptomatic therapy. The Ministry of Health issued directives to all provincial Centers for Disease Control and Prevention in May on monitoring, case investigations, and contact tracing. As winter sets, WHO is ramping up its coordination with the Ministry of Health to assure that the healthcare staff is equipped with the essential skills to address rising demands (https://moz.gov.ua/article/news/v-ukraini-zareestrovanij-vipadok-vispi-mavp).Currently, as an emergency is declared in Ukraine, after the 6-month war, the country is badly affected economically and socially. The COVID-19 pandemic is under surveillance in Ukraine, however, another epidemic monkeypox is reported in Ukraine on 15 September 2022. Ukraine has declared no historical case of MPXV case in its history as per Centers for Disease Control and Prevention (CDC). According to the announcement of the health ministry, the Ukrainian health system is taking lessons in providing life-saving care with the support of WHO and partnered for combating these challenges ahead. Ukraine has reported a total of five confirmed cases of monkeypox as of 15 October 2022 (The Russo-Ukrainian conflict has resulted in several sociopolitical, financial, infrastructural, and healthcare implications. Furthermore, the consequences of this conflict, particularly in terms of medical care including inside and even outside Ukraine, will persist long after the actual war has finished. It is critical to acknowledge that the Ukrainian medical sector is already experiencing considerable stress because of the increasing cases of casualties and harmful ramifications for the contemporary economy. These vulnerabilities are likely to exacerbate the spread of certain other infections including measles, typhoid, malaria, polio, HIV, and further stimulate the outbreak of the monkeypox virus. Overcrowded refugees in confinement camps have been implicated in the fast development of such illnesses. Moreover, interruptions in vaccination campaigns in conflict zones might serve a significant part in the emergence of other contagiousNot applicable.None.S.M. designed the study, made the first draft, updated the manuscript, and reviewed the final draft.The authors declare that they have no financial conflict of interest with regard to the content of this report.Not applicable.Sumira Malik."} +{"text": "This article was migrated. The article was marked as recommended.Due to the increasing number of COVID-19 cases globally, and the need for critical containment, Duke-NUS Medical School, Singapore, moved all of its preclinical classes online, keeping with national and university guidelines. The sudden move from face-to-face to online learning posed several challenges to the school\u2019s team-based learning (TBL) pedagogy. In TBL, student engagement is key to promote peer-to-peer learning. The educational faculty found that it was challenging to ensure student engagement through an online platform. Additionally, online TBL is heavily dependent on the use of technology. Technological and internet connectivity issues were potential obstacles to the learning process. This manuscript proposes practical tips for a facilitator of an online TBL class to engage learners in this new format. To overcome technical complications, a dedicated centralized administrative team managed the logistics of hosting TBL online. Working synergistically, the facilitator, and the administrative team were instrumental in recreating the learning environment of a face-to-face TBL in an online platform. The rapid transitioning to online learning made it difficult to ensure all participants were fully prepared beforehand. Being forced to engage remotely, it was difficult for participants to receive the technical assistance that they would otherwise receive in person.We adopted several different strategies to enhance the dynamism and sustain the engagement of students in online TBL sessions. Many of these revolved around consciously maintaining open forms of communication between the students and the faculty, and are as follows:1.We would communicate the sequence of events and the facilitation strategy to our students and faculty at the beginning of every facilitated discussion session. This allowed the participants to mentally prepare for each phase of the discussion. Priority and extra time would be allocated to questions that were poorly attempted, as well as to topics corresponding to critical learning points , Office of Education, Duke-NUS Medical School, and an Assistant Professor in the Neuroscience and Behavioural Disorders Program, Duke-NUS Medical School and Department of Pharmacology, National University of Singapore; ORCID ID:https://orcid.org/0000-0003-3734-2461.Muhammad Raihan Jumat is a Basic Science Education Fellow, Office of Education, Duke-NUS Medical School; ORCID ID:https://orcid.org/0000-0002-5098-6707.Irene Cheng Jie Lee is an Assistant Professor, Office of Education, Duke-NUS Medical School; ORCID ID:https://orcid.org/0000-0001-6862-6438.Ke Xiang Foo is an Assistant Manager of MD Programme Administrative Team, Office of Education, Duke-NUS Medical School.Suzanne Pei Lin Goh is Associate Dean of Student Affairs, and facilitator for Body and Disease Course, Office of Education, Duke-NUS Medical School.Sashikumar Ganapathy is an Assistant Dean, Clinical Integration, Office of Education, Duke-NUS Medical School and Deputy Head & Consultant, Emergency Medicine, KKH, Singapore; ORCID ID:https://orcid.org/0000-0001-5051-4542.Siang Hui Lai is a Senior Consultant with the Department of Anatomical Pathology, Singapore General Hospital, Editor of Proceedings of Singapore Healthcare, SingHealth, Academic Vice Chair (Education), Pathology Academic Clinical Programme, SingHealth Duke-NUS AMC, and Associate Professor and Associate Dean, Office of Education, Duke-NUS Medical School.Nian Chih Hwang is a Senior Consultant Anaesthesiologist with Singapore General Hospital and National Heart Centre, Singapore, and the Course Director of Body and Disease, Office of Education, Duke-NUS Medical School; ORCID ID:https://orcid.org/0000-0002-1220-345X.The author has declared that there are no conflicts of interest.Since no personal information was collected, ethical approval was not required for this manuscript.This article has not had any External Funding This review has been migrated. The reviewer awarded 5 stars out of 5 Very informative and detailed Thank youReviewer Expertise:NANo decision status is available This review has been migrated. The reviewer awarded 4 stars out of 5 Thank you for the opportunity to read and review this manuscript. The authors have systematically described best practices for a TBL curriculum that was delivered online due to the Covid-19 pandemic. The need for the innovation is sound, description of the intervention is clearly written such that others can adopt and adopt these best practices. The authors have also categorized the article under challenges, best practices for facilitation and how support was provided for teachers and learners.The manuscript is well written, well referenced and has clarity.I would have liked to have seen the following:(1) How faculty development was provided (2) How buy-in from teachers and learners was ensured. Was there a needs assessment, did they confer with these stakeholders?(3) A brief description of the impact of this curriculum- 2 levels of Kirkpatrick (reaction and knowledge) could have been evaluated briefly. I understand the transition was rapid, unexpected and has not been rolled out for very long.With these details and perhaps a theoretical underpinning of the benefits of TBL would make this a high quality scholarly manuscript.In its current form, it is a very good faculty development resource and informs other educators engaged in TBL courses about challenges and best practices.Reviewer Expertise:NANo decision status is available This review has been migrated. The reviewer awarded 4 stars out of 5 An important, informative experience for adopting online TBL using the official LMS. The challenge of students' engagement is the same all over the world. In our institution we adopted online TBL in a class of about 900 students. But we decided a different approach. As we stuck to our national official LMS for only the educational materials and processes with copyright requiring high degrees of security and used the commercial, free, available platforms in a portfolio of tools to provide a high degree of accessibility. We used the zoom with the breakout rooms to divide the students into teams of nine with the facilitator navigating throughout the rooms. This approach ensured the students' engagement and a close supervision. I greatly appreciate the opportunity for reading your experience. Our contribution is attached below. I hope to read more experiences from your renowned Institute. Dalia A. Gaber Mohamed Hany Shehata Hebat Allah A. Amin. (2020). MEDICAL EDUCATION ADAPTATIONS Online team\u2010based learning sessions as interactive methodologies during the pandemic. Medical Education. Volume 54, Issue 7. First published:26 April 2020https://doi.org/10.1111/medu.14198Reviewer Expertise:NANo decision status is available This review has been migrated. The reviewer awarded 4 stars out of 5 This is an interesting article on team-based learning and the rapid changes which had to be undertaken while shifting TBL to an online mode during the COVID-19 pandemic. The authors provide tips which would be useful to educators in other settings. Singapore being a developed high-income country some of the author\u2019s suggestions may need to be modified while applying them to resource limited settings. The major challenge in online learning is keeping the student engaged and focused. There are a number of challenges in achieving this as has been mentioned in this and other studies. Another challenge for the teacher/facilitator during online delivery is gauging the involvement of students. TBL incorporates certain elements like the readiness assurance tests and involvement in a group which encourages attention and participation. The article will be of broad interest with the increasing adoption of TBL as a learning method.Reviewer Expertise:NANo decision status is available"} +{"text": "This is despite a higher infection rate (p < 0.0001) in the non-Black population. The median household income of a zip code was negatively correlated with COVID-19 hospitalizations , but did not correlate with infections and deaths. Conclusions: The current study demonstrates clear health disparities of income and race in the context of COVID-19-related infections and outcomes in the city of Gary. Indiana University School of Medicine Northwest and GHD officials can collaborate to utilize these data for the reallocation of resources and health education efforts in Gary\u2019s highly populated, low-income, and predominantly Black neighborhoods. It should also prompt further investigation into national health resource allocation.Background: The COVID-19 pandemic further exposed the prevalence of existing health disparities in Black communities in the U.S. The current study evaluates COVID-19 data collected in Gary, Indiana, from June 2020 to June 2021. We hypothesized that the number of COVID-19 cases, hospitalizations, and deaths were influenced by race and income. Methods: In collaboration with the Gary Health Department (GHD), we analyzed demographic data on COVID-19-positive cases. Results: Compared to Gary\u2019s non-Black population, age- and population-adjusted rates of hospitalizations and deaths in the Black population were 3-fold ( The COVID-19 pandemic had a devastating effect on numerous countries across the globe. It put a severe strain on health facilities and economies . There iSocial factors that exist in Black and other minority communities, such as historically based mistrust of medical professionals, are also crucial to consider . In ordePrevious studies support the idea of a Matthew effect by which pandemic instability widens inequality by disproportionately affecting groups that are already disadvantaged . In addiGiven the multi-factorial nature of health disparities, it is imperative to assess them at all levels ranging from local communities to a national scale. Documenting and evaluating the racial and economic disparities related to COVID-19 in particular is crucial for improving healthcare policy, assistive programs, and resource allocation at the local, state, and federal levels of affected communities. The focus of this study is to analyze the effects of COVID-19 specifically in the city of Gary, Indiana. Gary serves as a microcosm for impoverished, majority non-White, and urban communities across America. There is a large Black population, a high number of residents living below the poverty line, and a high unemployment rate . The citAccording to the 2020 Census, the total population in Gary is 69,093 people and 78% of the population is Black, compared to 13% nationally. The mean household income of Gary residents is USD 34,085, which is well below the national average of USD 69,717 [COVID-19 data were collected from the Gary Health Department from 16 June 2020 to 7 June 2021. De-identified data from all 5149 positive cases included demographics and indicated disease progression . Demographic data on Gary and its zip codes, including population, age distributions, racial distribution, and median household income, were collected from the U.S. Census Bureau [COVID-19 data from GHD were re-binned to match Census age groups. African Americans in the city of Gary represent 79.8% of the population as compared to the 14.1% non-Hispanic White Americans. Since races other than Black are underrepresented in Gary, COVID-19 racial data were binned to include Black and non-Black. For the purpose of this study, non-Black includes Other, White, Multi-Race, Asian, and American Indian/Alaska Native.COVID-19 cases with missing race data were omitted for the racial differences analysis. Omitted data represented 46.7% of cases. This issue is not unique to the current study since race is underreported in 35% of COVID-19 cases nationally .Observed/Expected (O/E) ratio normalization for the age distribution effects of COVID-19 prevalence and severity were calculated and adjusted for missing race data using the following formula:O/E (observed/expected):O/E ratio effectively adjusts for population size, age distribution, and data with missing race. The O/E ratio is a commonly used method of data comparisons in the clinical setting .p-value of 0.05 or less. In this analysis, the expected values were calculated using the same formula for the expected count used in In the analysis of racial disparities, Pearson\u2019s chi-squared test was used to determine statistical significance, defined as a two-sided 2 to describe covariance in order to avoid issues with the model assumptions.In the analysis of the zip code distribution data, linear regression was used to model differences in COVID-19 severity and zip code-related variables. Linear regression analysis was used to determine the significance of differences in the population. Since these zip codes are adjacent to each other, the Pearson correlation coefficient was used instead of R2 to describe covariance in order to avoid issues with the model assumptions.Exponential linear regression analysis was used to determine the significance of income distribution by the zip codes on COVID-19 outcomes. O/E ratios were converted to a log scale, and then linear regression was performed on logged O/E ratios. Since these zip codes are adjacent to each other, the Pearson correlation coefficient was used instead of Rp < 0.0001; The analysis of the number of positive cases showed that there was a higher number of infections in the non-Black population compared to the Black population population size affected the number of positive cases; (2) racial disparities are present in the number of infections, hospitalizations, and deaths in the Black population versus the non-Black population in Gary; and (3) COVID-19 hospitalizations have a significant negative association with zip code income. The critical assessment and evaluation of these results can assist the Gary Health Department in creating a more targeted COVID-19 response to better address the needs of the Gary community. In addition to addressing COVID-19 impact, informed measures would also help to prevent the exacerbation of these disparities by future outbreaks.It is important to note the stark disparities in COVID-19 impact between the neighboring zip codes found in this study. Negative differences in health outcomes for neighboring residents are concerning for factors that can be addressed by local interventions, and zip code data may serve as a strong starting point to develop these interventions . As a mi"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Nature Communications 10.1038/s41467-023-40082-7, published online 19 July 2023Correction to: In this article the funding from \u2018the Korea Drug Development Fund, funded by the Ministry of Science and ICT, the Ministry of Trade, Industry, and Energy, and the Ministry of Health and Welfare \u2019 was omitted. The original article has been corrected."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.1As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.3Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.5Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.7Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.16Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.22Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.10In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "In this study, we aimed to evaluate the feasibility, utility, and potential effects of LQ-M/D App, a smartphone application developed by Life Quest Inc., Tokyo, Japan, for patients with mild cognitive impairment (MCI) and mild dementia. The app incorporates cognitive and physical exercise training, lifestyle habit acquisition features, and a continuity improvement feature added in the post-update version to enhance user engagement. The continuity improvement feature includes the optimization of training content, and disease education, and enables family monitoring via a family app.A retrospective analysis was conducted on app usage, cognitive and exercise training implementation and interruptions, questionnaire response rates, and cognitive assessments in a single institution. A total of 20 patients used the app, with 10 patients using the pre-update version without the continuity improvement feature, and the other 10 patients using the post-update version with the continuity improvement feature.The results demonstrated that the LQ-M/D App could be effectively used by the study population, and the continuity improvement feature positively influenced app usage in several aspects. Although a potential association between app usage and cognitive ability was suggested, the scatter in the data points warrants cautious interpretation. Limitations of the study included a small sample size, a single institution setting, and the retrospective nature of the study. In the future, a randomized controlled trial design using a larger sample size and multiple institutions to further evaluate the effectiveness of LQ-M/D App in managing MCI and mild dementia should be performed. The prevalence of dementia in Japan is increasing, and the number of people in Japan with dementia in 2025 is estimated to be 6.75 million . AlthougIn recent years, the field of mobile health using smartphones and mobile phones has gained attention, and is beginning to be used for older adults , 13, incAs described above, it has been reported that providing exercises, scheduling, and obtaining activity records via smartphones can be used without problems by older patients, including those with cognitive impairment. In addition, computer-based cognitive training can be substituted by mobile devices, such as smartphones. A smartphone application for patients with MCI or mild dementia called \u201cLQ-M/D App\u201d was developed by Life Quest Inc. , which includes the following functions: (i) training content (cognitive training and exercise training), and (ii) content for the acquisition of lifestyle habits . At present, there are a limited number of treatment-focused apps specifically tailored for patients with MCI or dementia. The LQ-M/D App is specifically designed for older adults with cognitive impairments to provide a non-pharmacological intervention. The purpose of this study was to investigate the feasibility and utility of the LQ-M/D App, and we conducted a retrospective observational study to analyze the clinical and application data of patients with MCI or mild dementia who used LQ-M/D App.This observational study was retrospectively conducted to investigate the effects of LQ-M/D App on patients with MCI or mild dementia treated at Hirokawa Clinic, Kyoto, Japan, from September 1, 2020, to August 31, 2021, using the clinic's medical records and data from the app. All patients were provided with smartphones containing the pre-downloaded LQ-M/D App, and used the LQ-M/D App, as well as received standard treatments for MCI or mild dementia. All procedures were performed in accordance with the ethical principles described in the 1995 Declaration of Helsinki. Written informed consent was obtained from each patient. The study was approved by the Institutional Review Board of Juntendo University and registered in UMIN Clinical Trials (UMIN000047077). In this study, external monitoring by the Japan Organization for Research and Treatment of Cancer (JORTC) verified data accuracy by comparing original sources with analysis data for a random subset of cases, with no discrepancies identified, confirming data integrity.In this study, the smartphone application \u201cLQ-M/D App\u201d developed by Life Quest Inc. was used. LQ-M/D App has two main features: (i) provision of training content, and (ii) content for the acquisition of lifestyle habits .The training content consists of cognitive function training and physical exercise training. The cognitive training consists of multiple menus targeting various cognitive functions, including memory function, language function, judgment function, calculation function, executive function, and visuospatial cognitive function, with a total of 21 different tasks. The physical exercise training comprises dual-task training and other types of training, and includes 38 dual-task exercises and 24 other types of physical activities. The lifestyle habits acquisition feature collects information on diet, sleep, and walking time through questionnaires conducted within the app. However, at present, the app does not have a feature for physicians to review records and provide feedback.On December 21, 2020, LQ-M/D App was updated to include a continuity improvement feature designed to enhance sustained user engagement. This feature consists of optimizing training content, providing disease education, and enabling family monitoring via a family app . The optIn this study, we used both the pre-update version without the continuity improvement feature, and the post-update version with the continuity improvement feature, each for a duration of 6 months, to compare and investigate their respective effects. Patients and caregivers were provided with comprehensive instructions on how to use the app. During regular monthly check-ups, healthcare providers asked about app usage to evaluate its use by the patients and caregivers. However, the actual usage of the app was ultimately at the discretion of the users themselves.In this study, data from medical records were retrospectively collected, including patient background, MCI and dementia treatment courses, and scores for Mini-Mental State Examination (MMSE), Hasegawa's Dementia Scale-Revised (HDS-R), Clinical Dementia Rating (CDR), and Alzheimer's Disease Assessment Scale-Cognitive Subscale Japanese version (ADAS-jcog). Additionally, information on app malfunctions occurring during use, daily app usage duration (in minutes), the daily number of cognitive training and exercise training sessions performed, daily number of interruptions of cognitive training and exercise training, and the presence or absence of daily questionnaire responses were obtained from the app data. The data were compared between the pre-update version without the continuity improvement feature and the post-update version with the continuity improvement feature. Furthermore, MMSE scores of 24\u201327 points were classified as MCI, and those of 20\u201323 points as mild dementia. For these patients, the association between daily app usage duration (in minutes), the daily number of cognitive training and exercise training sessions, and changes in MMSE, HDS-R, and ADAS-jcog scores from the start of app use to 6 months after the end of app usage were examined.t-tests were conducted for daily app usage duration (in minutes), daily number of cognitive training and exercise training sessions, and daily number of interruptions for cognitive training and exercise training. Additionally, the Mann-Whitney U-test was performed to compare the presence or absence of daily questionnaire responses between the pre-update and post-update versions. Pearson's correlation coefficients were measured between the changes in MMSE, HDS-R, and ADAS-jcog scores and daily app usage duration, and daily number of cognitive training and exercise training sessions. Standard deviations of mean dataset values were calculated. All p-values were two-sided, and a p-value of less than 0.05 was considered to indicate a statistically significant difference between two groups. All statistical analyses were performed using Easy R software , a graphical user interface that is a modified version of R .A total of 20 patients (7 men and 13 women) used LQ-M/D App, with a mean age of 72.3 (range: 50\u201382) years at the start of app usage. The MMSE, HDS-R, CDR, and ADAS-jcog scores at the start of app usage are shown in During usage of the app, 11 patients experienced malfunctions. In 4 cases of patients using the pre-updated version, the training feature of the app was unstable for 36, 34, 26, and 8 days, respectively. In 6 cases of patients using the post-updated version, the app could not be launched owing to an inability to communicate with the server for 14, 13, 12, 8, 6, 5, and 4 days, respectively.n\u2009=\u200910) was compared with the post-update version with the continuity improvement feature (n\u2009=\u200910). For each version, the mean daily app usage time, mean number of cognitive training and exercise training sessions, mean number of interruptions, and presence or absence of questionnaire responses were shown for the entire period and from the first to the sixth month, in In this analysis, the pre-update version without the continuity improvement feature , as well as for the first and fourth months. On the other hand, at 6 months, the number of sessions decreased in the pre-update version, and a significant increase in the number of sessions was observed in the post-update version . For theThe mean daily questionnaire response rate tended to be lower for the post-update version, except for the sixth month. A significantly higher mean questionnaire response rate was observed for the pre-update version during the overall 6-month observation period, as well as at the first, second, and fourth months . Howeverp-values were not statistically significant, suggesting that further research is needed to confirm these findings and their effects on cognitive function.In this study, patients with MMSE scores of 24\u201327 were classified as having mild MCI, and those with scores of 20\u201323 were classified as having mild dementia. Four patients with MMSE scores of 28 or higher and 1 patient with a score of 19 or lower were excluded. The remaining sample comprised 11 patients with MCI and 4 with mild dementia, totaling 15 patients. The association between daily app usage duration (in minutes), the daily number of cognitive training and exercise training sessions, and changes in MMSE, HDS-R, and ADAS-jcog scores from the start of app use to 6 months after the end of app use was investigated for these 15 patients using Pearson's correlation coefficients. Scatterplots of the associations are shown in In this study, we investigated the feasibility and utility of LQ-M/D App, a smartphone application developed for patients with MCI or mild dementia. The results demonstrated that the app could be utilized by the study population without major issues, and that the continuity improvement feature introduced in the updated version of the app appeared to positively influence app usage in several aspects. Although some app malfunctions were observed during the study, they were within acceptable limits and did not greatly hinder the app's usability. However, although the association between app usage and cognitive ability of the patients showed a tendency for a positive correlation, the available data was not sufficient to draw definitive conclusions. Therefore, further research is needed to confirm these findings and their effects on cognitive function.Previous studies have reported the potential benefits of nonpharmacological interventions, such as moderate physical activity and lifestyle modifications, in slowing cognitive decline and reducing the progression from MCI to dementia , 24. LQ-Our study observed a generally low questionnaire response rate in both the pre-update and post-update versions of LQ-M/D App. A possible reason for this might be the lack of evaluation and feedback on patients\u2019 questionnaire responses, as feedback has been shown to improve engagement and adherence in eHealth interventions . The posp-values. These findings suggest a potential association between app usage and cognitive improvement but should be interpreted with caution owing to the small sample size and the retrospective nature of the study. Moreover, among the 20 patients, 17 were receiving medication for dementia, and 2 patients started taking medication during the study period. Therefore, the possibility of the slight improvement in cognitive performance being a result of the effects of the medication rather than the effects of training using the App cannot be ruled out. Further research with larger sample sizes and prospective study designs is necessary to better understand the effects of the LQ-M/D App on cognitive function in patients with MCI or mild dementia, and to confirm the potential cognitive benefits of the app and to investigate the underlying mechanisms contributing to these correlations, as suggested by prior research on the effectiveness of cognitive training for cognitive function (In this study, the association between app usage and cognitive improvement did not reach statistical significance, with correlation coefficients demonstrating wide-ranging confidence intervals and relatively high function .There are some limitations to our study. First, the sample size was relatively small, which may have affected the statistical power of the analyses. Second, the study was conducted in a single institution, which may limit the generalizability of the findings. Third, the study was retrospective in nature, which may have introduced biases in the data collection and interpretation. Fourth, the study lacked a control group, making it difficult to determine the specific effects of the LQ-M/D App on cognitive function compared with standard treatments alone. Future research should consider a randomized controlled trial design with larger sample sizes and multiple institutions to further evaluate the effectiveness of LQ-M/D App in managing MCI and mild dementia.In conclusion, this study demonstrated the feasibility and potential utility of LQ-M/D App for patients with MCI or mild dementia. The app's continuity improvement feature appeared to positively influence app usage, and the observed trends in cognitive assessments suggested potential cognitive improvement, although further research is needed to confirm these findings. LQ-M/D App may be a promising tool for the management of MCI and mild dementia, potentially contributing to the prevention or delay of cognitive decline in this population."} +{"text": "The coronavirus disease 2019 (COVID-19) pandemic has precipitated the adoption of online teaching-learning methodologies at all educational levels worldwide, affecting all types of training . This has given rise to a seemingly new debate on the effectiveness of training that the call for face-to-face training has provoked. However, this debate, apart from the online face-to-face dilemma or their combination, is familiar.The problem of the transfer of training has been of concern to both academics and practitioners. However, only a small percentage of what is learned during training applies to jobs. Consequently, there is a paradox, an explanatory gap in the relationship between training and performance. With the transfer of training, training efforts can contribute to organizational effectiveness , depreciation of training, technological advances, and issues of equality, diversity, and inclusion are also considered.Yahiaoui et al. investigated the impact of total quality management practices in higher education institutions; S\u00e1nchez-Prieto et al. and Procopio et al. focused on novel approaches in higher education and more active and participatory teaching innovation methods. The authors presented a competitive debate regarding gamification among teams with university students, a didactic experience of implementing a methodological approach based on cooperative learning, and a review of the literature on neuroeducation. Gradellini et al. examined the content and knowledge of cultural competence and intercultural communication offered in higher education in nursing courses. Barba-S\u00e1nchez et al. developed a model for assessing the impact of secondary school information technology (IT) capacities on smart city business development based on the IMD Smart City Index, PISA, and World Bank reports. G\u00f3mez-Cantarino et al. presented a narrative review of health education models, conceptual frameworks, and the importance of information and communication technology (ICT) in a transdisciplinary approach to child abuse. The impact of human resource management practices was discussed by Xie et al., where the impact of human resource management practices and training and development types in multinational enterprises was considered; Xiao et al. studied how supervisors' developmental feedback affects creativity at the team level. Noman et al. examined the moderating role of cross-cultural training in the adjustment of self-initiated and organizational expatriates. Based on dynamic capability theory, Wang et al. researched the effect of a high-performance work system on organizational performance, the mediating role of strategic flexibility, and the moderating role of an enterprise's social network in this relationship. Zhiqiang et al. examined how high-performance work practices (HPWPs) affected employees' in-role performance (EIRP) and task performance (ETP) during the COVID-19 pandemic.The papers selected in this research address the latest research framed in new approaches to training and education from the perspective of public policy, prospective organizations, and individuals. Considering the role of higher education, Peng et al. conducted a bibliometric study on innovation research in organizations within the three levels using the CiteSpace software to analyze 6,354 academic articles from the year 2000 to 2020. Respect for ecology within organizations is important in both training and education. Baeshen et al. discussed a study to develop a comprehensive model by integrating the natural resource-based view (NRBV) and triple bottom line (TBL) framework. The influence of absorptive capacity and innovativeness on business performance was analyzed by Sancho-Zamora et al., considering the relationships between the different dimensions of absorptive capacity and innovativeness.Understanding the state of research on organizations is fundamental to this special issue. This research combines high-quality research with different quantitative and qualitative methodologies and mixed-methods studies. It also includes reviews that follow quality criteria, and research projects that adopt multisectoral and transdisciplinary approaches.BY-A, JM-G, and FH-P contributed to conception and design of the study and wrote sections of the manuscript. BY-A wrote the first draft of the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version."} +{"text": "This is a peer-review report submitted for the paper \"COVID-19 Testing Strategies and Lockdowns: The European Closed Curves, Analyzed by Skew-Normal Distributions, Forecasts for the United Kingdom, Sweden, and the United States, and the Ongoing Outbreak in Brazil.\"This paper is very"} +{"text": "Ilona Kurnatowska and Irena Maniecka-Bry\u0142a were not included as authors in the original publication . The corAn Affiliation of Ilona Kurnatowska is added: Affiliation 2: Department of Internal Medicine and Transplant Nephrology, Medical University of Lodz, Kopcinskiego 22, 90-549 Lodz, Poland.Author Contributions: P.P.-G., contributed to the study design and writing the article; I.K., contributed to the study design; I.M.-B., contributed to the study design; M.P., contributed to the study design and writing the article, and conducted the statistical analysis and interpreted the data. All authors have read and agreed to the published version of the manuscript."} +{"text": "Nature Communications 10.1038/s41467-023-36553-6, published online 16 February 2023Correction to: This Article contains an error in the name of the author Wei Chou Tseng, which is incorrectly given as Wei Tseng. This has been corrected in both the PDF and HTML versions of the Article.In the original version of this Article, the affiliation details for Buchmann Institute for Molecular Life Sciences and Institute of Biochemistry II, Faculty of Medicine, Goethe University Frankfurt, Frankfurt, Germany were incorrectly given as Buchmann Institute for Molecular Life Sciences, Goethe University Frankfurt, Frankfurt, Germany. This has now been corrected in both the PDF and HTML versions of the Article.The original version of this Article contained an error in the Acknowledgements which incorrectly read \u2018the Collaborative Research Center SFB 1177 (ID 259130777 (C.B.)\u2019 The correct version states \u2018\u2019 in place of \u2018(C.B.)\u2019. This has been corrected in both the PDF and HTML versions of the Article."} +{"text": "Dr. Pereira-Sanchez will discuss how the climate change emergency apeals psychiatrists and demands both personal and organized responses. Such responses are in the domains of awareness, research, education, and action. Dr. Pereira-Sanchez will present specific examples from his collaborative work at the World Network of Psychiatric Trainees, where a global forum about the topic for trainees was organized, and the World Psychiatric Association, where he assists the coordination of a Tri-Sectional initiative on the topic.None Declared"} +{"text": "The link between geography and health means that the places we occupy\u2014where we are born, where we live, where we work, and where we play\u2014have a direct impact on our health, including our experiences of health. A subdiscipline of human geography, health geography studies the relationships between our environments and the impact of factors that operate within those environments on human health. Researchers have focused on the social and physical environments, including spatial location, patterns, causes of disease and related outcomes, and health service delivery. The work of health geographers has adopted various theories and philosophies and methods to collect and analyze data to examine our environments and their relationship to health. The field of public health is an organized effort to promote the health of its population and prevent disease, injury, and premature death. Public health agencies and practitioners develop programs, services, and policies to promote healthy environments to support and enable health. This commentary provides an overview of the recent landscape of health geography and makes a case for how health geography is critically important to the field of public health, including examples from the field to highlight these links in practice. Health geography studies how human health and health systems are diffused, distributed, determined, and delivered by using a spatial lens to examine these factors across a range of scales ,2. IndeeThe Canadian Public Health Association defines public health as \u201cthe organized effort of society to keep people healthy and prevent injury, illness and premature death\u201d . Public Health geographers have made important contributions to public health scholarship on the reciprocal relationship between place and human health. Most notably, geographers have acknowledged that contextual, compositional, and collective aspects of place together influence population health in a much more relational rather than unilateral way ,6. The iThe factors and conditions impacting health status are also known as the determinants of health. It is estimated that factors such as behaviour, biology, and access to and quality of healthcare only determine about half of an individual\u2019s health outcomes. The other half of these outcomes are determined by the conditions in which people are born, grow, work, live, and age. These conditions are known as the social determinants of health and include factors such as income, education, working conditions, race, gender, and culture .In Canada and worldwide, there are historical, persistent, and emerging health inequities. The term inequity is used to: \u201cdescribe those health inequities, though avoidable, are not avoided and hence are unfair\u201d , Avian influenza (H5N1)) [There is growing concern about public health issues impacting people\u2019s health at the population level. According to the World Health Organization (WHO) health is a fundamental right, defined as \u201ca state of complete physical, mental and social well-being and not merely the absence of disease or infirmity\u201d (p. 1). (H5N1)) ,57.The social determinants of health (SDOH) refer to specific factors outside of the healthcare system or healthcare sector that influence the health of individuals and populations and reflect one\u2019s location in society by way of income and social status, race, gender, education and literacy, employment status, and working conditions . The SDOThere have been recent calls to re-examine and continue expanding the SDOH, since they omit structural racism and the health system itself as a SDOH . This caStructural determinants of health and the root causes of inequities have been examined by Crear-Perry and others . Specifihttps://ncceh.ca/resources/blog/renewed-attention-environmental-equity-and-justice (accessed on 12 June 2023)). Health geographers have promoted environmental justice initiatives and approaches and advocated for the inclusion of community voices to better inform the development of environmental health public policy, multiscalar analysis, and interdisciplinary partnerships [Significant attention has been paid by health geographers to environmental injustice. This includes the actions and policies that place groups or communities, regardless of social position, at greater health risk through increased proximity and exposure to hazardous environmental conditions or natural disasters . FurtherIn the 1990s, a shift from medical to health geography included a recognition of the need to explore health and illness in the context of experiences of being in place, defined as the local environment in which social process, health, and disease occur ,79. A foAlong with this disciplinary shift, health geographers expanded their focus beyond largely positivist approaches to those informed by theory and more qualitative in nature ,81. As sA shift towards focused efforts on public health initiatives in health geography has identified health and place as critically important for engagement between health geography and public health . Health An overarching goal of health promotion and disease prevention is to support, and not hinder, healthy behaviour. A focus on the environment, including un/supportive environmental factors, has included research on the relationship between individual behaviours and features of the built, social, and policy environments at the local level to examine immigrant well-being ,86, chilhttps://www.toronto.ca/community-people/health-wellness-care/health-programs-advice/respiratory-viruses/covid-19/covid-19-pandemic-data/covid-19-vaccine-data/ ; City of Hamilton (cases): https://www.hamilton.ca/people-programs/public-health/diseases-conditions/coronavirus-covid/covid-19-data#incidence-rate-by-ct (accessed on 3 June 2023). These data were used to inform vaccine clinic planning as well as neighbourhood-level vaccine uptake strategies. The City of Toronto, Canada collects data on neighbourhoods to address planning needs and the social determinants of health. As such, Toronto developed a neighbourhood map (N = 158) to track the weekly dose count of the population vaccinated for COVID-19 during specific periods according to neighbourhood of residence. As such,Inequities in health were examined during the COVID-19 pandemic, linking inequities in COVID-19 outcomes with existing inequities in the social determinants of health and chronic diseases ,100. A rHealth inequity can explain why some neighbourhoods may have lower vaccination rates. Health inequity refers to preventable differences in health between groups of people \u2026 Inequities in vaccination uptake are often a result of lack of availability of services in one\u2019s neighbourhood, lack of flexible service hours and access to transportation to attend vaccine clinics\u2026 In addition, lower vaccine uptake has been linked to negative historical experiences with health care institutions and distrust of health care, much of which stems from discrimination, systemic racism, and the effects of colonization (p. 5).Population Health Intervention Research (PHIR) seeks to develop and evaluate interventions (policies and programs) that influence health at the population health level while considering health equity and the contexts in which interventions are designed, established, implemented, and evaluated . These ihttps://www.publichealthontario.ca/en/data-and-analysis/commonly-used-products/snapshots (accessed on 2 June 2023). Public Health Ontario and researchers at the MAP Centre for Urban Solutions at St. Michael\u2019s Hospital have also developed the Ontario Marginalization Index (ON-Marg) ), which measures and maps differences in marginalization across Ontario. Data are available at multiple levels, including at dissemination areas and public health unit levels) [Topics of interest to health geographers in PHIR have included housing as an important setting for improving health, such as in the contexts of improvements to safety, tenant interactions with a house, material conditions and mental health, and the symbolism and meaning of being housed after homelessness ,109. Ano levels) .More generally, the built environment has seen a strong focus in health geography using geocoding , metropolitan areas, divisions, economic regions, federal electoral districts, population centres, census tracts, and dissemination areas) , and Onthttps://resources.esri.ca/news-and-updates/the-2023-university-of-toronto-geospatial-data-visualization-challenge (accessed on 6 May 2023)). ArcGIS developed a COVID-19 Health Dashboard to produce maps illustrating the number of COVID-19 cases by province and confirmed cases by provincial public health units, and population density at the dissemination area and neighbourhood level (https://resources-covid19canada.hub.arcgis.com/apps/90fdd2da4bba4c79a33c2202760b3c5d/explore (accessed on 6 May 2023)).The Dalla Lana School of Health at the University of Toronto recently hosted a 2023 Geospatial Data Visualization Challenge, whereby teams developed ArcGIS story maps to present a public health issue of choice through maps and visualization to highlight child poverty and food insecurity, the opioid crisis, physical activity and well-being (The impacts of the COVID-19 pandemic in Canada and around the world are unprecedented and include the worsening of pre-existing health inequities ,100,102.At a larger scale, health geographers are well positioned to address health system strains and health human resource challenges in the Canadian context by undertaking additional research at the interface between public health and health services; for example, into the practice of social prescribing, or community-led referral to services to address needs rooted in the social and environmental determinants of health . Health Finally, health geographers have the potential to fill a leadership role with respect to knowledge integration across scales and disciplines necessary to address the impending public health challenges. The increasing recognition of systems complexities in ongoing public health polycrises and syndemics , from th"} +{"text": "In \u201cThe Efficacy, Safety, and Efficiency of the Off-Label Use of Bevacizumab in Patients Diagnosed With Age-Related Macular Degeneration: Protocol for a Systematic Review and Meta-Analysis\u201d the authors noted a missing Acknowledgments section. The Acknowledgments section has now been added to the published paper as follows:H&TRC authors gratefully acknowledge the FCT/MCTES national support through the UIDB/05608/2020 and UIDP/05608/2020.The correction will appear in the online version of the paper on the JMIR Publications website on July 4, 2023, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "Instead of Frontiers Science Center for Disease-related Molecular Network, Sichuan University, Chengdu, China, it should be Institute of Respiratory Health, Frontiers Science Center for Disease-related Molecular Network, West China Hospital, Sichuan University, Chengdu, China.In the published article, there was an error in affiliation The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.The data presented in this study have been deposited in the Genome Sequence Archive (GSA) in BIG Data Center, Beijing Institute of Genomics (BIG) under accession number HRA003044 (https://ngdc.cncb.ac.cn/gsa-human/s/epL1lQWd).Additionally, in the published article, there was an error in the Data Availability statement. Instead of The correct Data Availability Statement appears below:The data presented in this study have been deposited in the Genome Sequence Archive (GSA) in BIG Data Center, Beijing Institute of Genomics (BIG) under accession number HRA003044 (https://ngdc.cncb.ac.cn/gsa-human/browse/HRA003044).The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.The funding statement for the Fundamental Research Funds for the Central Universities was displayed as \u201c20826041F4147. The correct statement is \u201cFundamental Research Funds for the Central Universities (No. 2022SCU12047 to Y. Chen)\u2019\u2019. The correct Funding statement appears below.Furthermore, in the published article, there was an error in the Funding statement. \u201cThis study was supported by National Natural Science Foundation of China , Post-Doctor Research Project, West China Hospital, Sichuan University (2021HXBH051), Sichuan Science and Technology Support Project (No. 2022NSFSC1516 to YC) and the Fundamental Research Funds for the Central Universities (No. 2022SCU12047 to Y. Chen)\u201d.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "Globally, there is a rise in chronic disease, including cancer, major organ failure and dementias. Patients and their families in low- and middle-income countries (LMICs) pay a high proportion of medical costs out of pocket (OOP), and a diagnosis of serious illness often has catastrophic financial consequences. We therefore conducted a review of the literature to establish what is known about OOP costs near end of life in LMICs.To identify, organise and report the evidence on out-of-pocket costs in adult end-of-life populations in LMIC.A systematic search of 8 databases and a hand search of relevant systematic reviews and grey literature was performed. Two independent reviewers screened titles and abstracts, assessed papers for eligibility and extracted data. The review was registered with PROSPERO and adhered to the Preferred Reporting items for Systematic Reviews and Meta Analyses. The Mixed Methods Appraisal Tool was used to assess quality. The Wagstaff taxonomy was used to describe OOP.After deduplication, 9,343 studies were screened, of which 51 were read and rejected as full texts, and 12 were included in the final review. OOP costs increased with advanced illness and disease severity. The main drivers of OOP were medications and hospitalizations, with high but variable percentages of the affected populations reporting financial catastrophe, lost income, foregone education and other pressures.Despite a small number of included studies and heterogeneity in methodology and reporting, it is clear that OOP costs for care near end of life in LMIC represent an important source of catastrophic health expenditures and impoverishment. This suggests a role for widespread, targeted efforts to avoid poverty traps. Financial protection policies for those suffering from incurable disease and future research on the macro- and micro- economics of palliative care delivery in LMIC are greatly needed. Low- and middle-income countries (LMICs) face sharply increasing incidence of non-communicable diseases such as cancer, major organ failure, and Alzheimer\u2019s disease and related dementias . Health The end-of-life phase has unique physical, psychological and spiritual challenges . There iPeople with terminal illness and their families in LMICs are vulnerable to bankruptcy and impoverishment as populations age, and the poorest sections of society are routinely the most vulnerable of all . One parFaced with impossible choices between earning income or caring for a loved one, or between retaining basic assets or paying for treatment, end-of-life care is a potential poverty trap for many . PalliatOut-of-pocket costs in LMICs are a long-standing policy question . The ecoWe are unaware of any prior review examining the costs to patients and families in the end-of-life phase in LMICs. We therefore conduct a review of the literature to establish what is known about out-of-pocket costs near end of life in those settings. Arising results can inform efforts in low-resource settings to upscale palliative care provision, to design financial protection policies for people with non-communicable disease, and to conduct future research on the economics of palliative and end-of-life care in settings where prior attention has been minimal.th, 2020.We registered a protocol for this systematic review on PROSPERO (CRD42020215188). Registration date was November 19A clinical librarian (AB) developed the search strategy after a consultation with ER and PM. AB also received related articles which helped formulate the search strategy with the use of the Yale MeSH Analyzer and were later used to validate search concepts.The search strategy was peer-reviewed by another senior librarian. The search strategy used both keywords and controlled and indexed vocabulary combining the terms for low-and-middle income countries, cost, and palliative care.The databases were searched from inception to August 5, 2020; the databases included: MEDLINE (Ovid), Embase (Ovid), APA PsycInfo (Ovid), Global Health (Ovid), CINAHL (Ebsco) Web of Science , Scopus, EconLit (ProQuest), and Cochrane CENTRAL. See We supplemented database searches with other methods. One reviewer (JJ) hand-searched three potentially relevant systematic reviews , 15, 16.Population. Adults (aged 18+) at the end of life or diagnosed with one of the following serious life-limiting medical illnesses: advanced cancer, serious heart disease , major organ failure , advanced Alzheimer\u2019s disease and related dementias, AIDS/HIV.Intervention. Palliative care, supportive care and end of life, opioids, and other pain management medications.Comparison. Any or none.Outcome. Out-of-pocket costs incurred by patients and their families in care and treatment for end-of-life care. We include studies that measure the monetary cost of providing informal care, including costed dedicated time, income foregone through lost work, and additional costs incurred .Setting. Low- and middle-income countries, as defined by the World Bank [rld Bank .Study design. Any.Population. Studies of children. Studies of people with other chronic diseases as a primary diagnosis. Studies of people in the early stages of a terminal disease.Intervention. Screening, identification, diagnosis of disease. Medications with a predominantly curative intent.Outcome. Costs of care where the patient does not contribute their own resources in money or time, e.g., any service free at the point of use. Prevalence of out-of-pocket payment or unpaid care where that prevalence is not quantified as an estimation of time or cost.Setting. Studies in high-income countries.Other. Studies where our population, outcome and setting are not specifically reported, e.g. (1) an overview of out-of-pocket costs at the population level that does not delineate those with terminal illness or at end of life, e.g. (2) a cost of illness study for a terminal illness that does not delineate out-of-pocket and centrally funded costs. Studies where out-of-pocket costs are hypothetical and/or employed as a predictor . Any publication type except research articles . We included only English language articles and retained eligible abstracts in other languages to assess risk of bias from the English-only constraint.We imposed three basic criteria: LMIC setting, palliative/end-of-life population and out-of-pocket costs as an outcome of interest. Among these, only the first can be categorized discretely according to objective rules (we use The World Bank) . For popEach title/abstract was screened as (in)eligible by two independent reviewers . Disagreements were settled by consensus, with either ER or PM acting as the third reviewer.Full texts were screened using the same process as titles/abstracts. Quality assessment of included studies was performed by two independent reviewers using the MMAT tool. There was no quality cut-off for inclusion, we resolved that eligible studies be reported in the context of their methods and limitations.Data were extracted using the same process as titles/abstracts: two independent reviewers, with a third senior reviewer (ER or PM) adjudicating any conflicts. Data were extracted to a bespoke form developed in Excel, that required data points on author, year of publication, country of data collection, year of data collection, study design, sample size , population, intervention/exposure, comparison, outcome of interest, main results, key themes/messages/strengths, and key limitations. The two reviewers\u2019 outputs were then merged into a single file by a third reviewer (AG or AK), and conflicts or problems solved by consensus.In anticipation of a small, heterogeneous literature we did not evaluate risk of bias specifically but assessed bias as part of quality assessment.In anticipation of a small, heterogeneous literature we did not pre-specify an outcome of interest but instead adopted a flexible approach depending on identified studies, which could include descriptive studies. We planned therefore for narrative synthesis, organizing reported out-of-pocket costs according to seven measures in a well-known systematic review: (i) expenditure in absolute terms; (ii) measures of dispersion (or risk); (iii) the out-of-pocket budget share; (iv) progressivity; (v) the incidence of \u201ccatastrophic\u201d expenditures; (vi) inequality in the incidence of catastrophic expenditures; (vii) the incidence of \u201cimpoverishing\u201d out-of-pocket expenditures, as well as the addition to the poverty gap due to out-of-pocket expenditures .The database search returned a total of 13,751 records with 9,337 unique articles. The hand search identified six additional studies and the grey literature search identified no relevant studies. These 9,343 studies were screened in Covidence. We excluded 9,280 articles after screening of titles and abstracts, and a further 51 articles after reading the full text. One of these exclusions was due to not being in English . Thus, wThe data extracted from the twelve included studies \u201333 are pThe earliest study was published in 2002 and the most recent in 2020 with a median publication year of 2018. There were six studies conducted in India, and one each conducted in China, Kenya, Pakistan, Tanzania, Thailand and Southeast Asia . See There were six descriptive cross-sectional studies, of which three were conducted prospectively , 27, 32 Six studies based their sampling frames on diagnosis of a specific disease: three in cancer, , 25, 29 Nine studies evaluated outcomes specified by the taxonomy of Wagstaff et al . Two stuQuality assessment were performed by two independent reviewers using the MMAT tool. The MMAT is intended to be used as a checklist for concomitantly appraising and/or describing studies included in systematic mixed studies reviews . There was no quality cut-off for inclusion, and studies have been reported in the context of their methods and limitations. A summary of the quality assessments is provided in Main results of each included study are presented in Tables Results with respect to financial catastrophe are summarized in In India Prinja et al. reportedKimman et al. followedResults with respect to costs in absolute terms are summarized in Three studies in the hospital setting found that OOP costs for deceased patients in inpatient care is much greater than that of survivors , 28, 31.Cancer costs in private health care settings were higher than those in public settings , and higOne study looked at unpaid carer burden, summarized in Studies quantifying the incidence or prevalence of financial burden in descriptive measures are summarized in These measures affirm the findings in other tables that advanced medical illness results in high costs and financial pressures. Further, illness and associated costs catalyze vicious economic circles for patients and families. Persons with serious illness overwhelmingly lose their jobs or leave the workforce , 33. FamPatients approaching end of life in LMIC pay a high proportion of medical costs out of pocket and often suffer catastrophic financial consequences, yet there is a dearth of robust, large scale health economic data to guide hypothesis-generating, poverty-reducing solutions. Financial catastrophe from serious medical illness is not only an issue for patients and families in the end-of-life stage but creates cyclical poverty traps: lost income, increasing borrowing and debt, anxiety and insecurity, and growing pressure to address financial pressures through drastic measures. Our systematic review directly addresses this global inequity with a rigorous search and summary of the existing literature, including a quality assessment.Strengths of the review include the broad search terms, inclusion of grey literature, the geographical heterogeneity in the included studies country of origin, and use of two independent reviewers at each step of the review. Limitations of this review are that much of the existing literature is from household surveys and interviews, thus introducing bias however this is mitigated by country-specific estimates for OOP which were included in three of our included studies. Only studies in English were included.Inadequate government spending on health is a recurring feature in LMICs. Weak health infrastructure leads to delays in diagnosis and resulting late disease presentations, heavy reliance on out-of-pocket payments and catastrophic health expenditures. Those patients impacted the most include those from remote areas, those with longer and repeat hospitalizations and with the following diagnoses: cancer, Alzheimer\u2019s disease, terminal HIV/AIDs and end stage liver disease. As out-of-pocket spending is inversely proportional to life expectancy, earlier diagnoses through improved screening programs would likely result in more treatable disease, at lower cost.Policies aimed at bolstering socioeconomic resilience and financial protection are greatly needed in LMICs. A better understanding of early versus late drivers of medical impoverishment is an urgent research priority, as this would inform these strategies. In addition to earlier detection of disease, the provision of early, home-based and widespread access to palliative care in LMIC would serve to decrease OOP and CHE for those with incurable disease, and thus should be considered as a critical health priority and global poverty-reduction strategy. Furthermore, in resource-scare environments, increased access to palliative care would have the secondary effect of liberating limited health resources for those patients with curable disease thus benefiting society at large. The need for greater access to palliative care in LMICs is clear and the evidence for its role as a poverty reduction strategy is emerging. Future research should focus on the implementation and health economic outcomes of palliative care in LMICs, thus improving care for billions of the most vulnerable patients in our global population.Globally, demographic ageing is likely to result in an increased burden on all levels of health systems and household services, partly due to the relatively long duration of illness at the end of life. Holistic palliative care can mitigate the desperate poverty caused by life-limiting illness, particularly if initiated early in the illness, on a regular basis and on a broad scale. Home-based treatment also frees up hospitals to serve patients with reversible conditions.Patients approaching end of life in LMIC pay a high proportion of medical costs out of pocket and often suffer financial catastrophe. These effects are long term and potential poverty traps: reduced household income, rising debt, deteriorating mental health, and narrowing life choices. Policies and interventions are needed to prevent these often-avoidable crises. Evidence to inform such policies and interventions is currently thin.S1 Checklist(DOCX)Click here for additional data file.S1 FileA text file of the search strategies used in our systematic review.(DOCX)Click here for additional data file.S2 FileFull details of the MMAT scores which are summarized in (XLSX)Click here for additional data file."} +{"text": "Cell Death Discovery 10.1038/s41420-023-01387-0, published online 05 April 2023Correction to: The original version of this article contained an error in the affiliations. Mao Benliang is affiliated with the College of Clinical Medicine, Guizhou Medical University, Guiyang, China and the Department of General Surgery, Guangzhou Red Cross Hospital affiliated to Jinan University, Guangzhou, China. The author also contributed equally to this article. The original article has been corrected."} +{"text": "The Japan Medical Association (JMA) has been engaged in supporting the health of employed physicians for several years. For example, an email and telephone consultation service has been provided since 2009, workplace improvement workshops have been conducted since 2010, and an analysis and improvement tool for labor management for physicians was published in 2014 (3). In accordance with the revised Labor Standards Act, physicians will be restricted to 960 hours of overtime per year from April 2024; however, they will be able to work up to an annual overtime limit of 1,860 hours under designated employment conditions (4). Despite this, the recent work schedules of Japanese physicians remain unclear, and events such as the coronavirus disease 2019 pandemic have dramatically changed the clinical situation (5). The current study aimed to provide an overview of trends in physician work schedules in Japan.Physician overwork is associated with depressive symptoms and suicidal ideation (6). In the current study, we excluded respondents aged 60 years or older, and for the analysis, we calculated descriptive statistics for each survey.This study used data from the first (2009), second (2015), and third (2021) nationwide employed physician surveys, which were repeat surveys conducted by the JMA. For each survey, self-administered questionnaires were mailed to 10,000 randomly selected employed physician members of the JMA. We obtained information about age, gender, monthly off-duty days, on-call days, and night shift days, and average sleeping hours on off-duty days from the dataset. Wada et al. previously published the study protocol and questions We included 3,067, 2,070, and 1,678 respondents in the analysis of the first, second, and third surveys, respectively . The per(7). In addition, because sleeping hours appear to be unchanged, self-study outside work hours should also be considered (4). Furthermore, lack of sleep is a risk factor for patient-related medical incidents (8) and may also harm physicians\u2019 health (9). Further efforts are required to comply with regulations regarding working hours.Our results suggest that physician work schedules in Japan showed some trends toward improvement from 2009 to 2021, although caution must be exercised due to the potential biases in sex, age, and response rate of the survey respondents. Notably, the number of off-duty days for employed physicians has been steadily increasing. Meanwhile, the number of on-call and night shift days slightly changed compared with the number of off-duty days. This gap may indicate that some physicians are engaged in on-call or night shift work instead of taking off-duty time NoneThis work was supported by the Japan Medical Association (no grant number).We thank all members of the Japan Medical Association who participated in or supported this study.TI: conceptualization, investigation, data analysis, and manuscript writing/editing. MO: investigation, data curation, and manuscript reviewing/editing. TY: project administration, investigation, and manuscript reviewing/editing. MK, YN and AN: investigation, and manuscript reviewing/editing. KK, SI, and KM: funding acquisition, supervision, and manuscript reviewing/editing.This study was approved by the Human Research Committee at the Institute for Science of Labour (2008-0020), the Ethics Committee of the University of Occupational and Environmental Health, Japan (H27-012), and the Research Ethics Committee of the National Institute of Occupational Safety and Health, Japan (2021N32). Participation was voluntary and anonymous; therefore, the requirement for written informed consent was waived.Kichiro Matsumoto is one of the Honorary Editors of JMA Journal and on the journal\u2019s Editorial Staff. He was not involved in the editorial evaluation or decision to accept this article for publication at all."} +{"text": "The contributions of this author are as follows: Conceptualization, Writing\u2013Review & Editing.Paula M. Roncaglia-Denissen should be included in the author byline instead of the Acknowledgments. Paula M. Roncaglia-Denissen should be listed as the third author, and their affiliation is 3: Department of Cognitive Science and Artificial Intelligence, Tilburg School of Humanities and Digital Scienceshttps://doi.org/10.1371/journal.pone.0229109The correct citation is: Sadakata M, Weidema JL, Roncaglia-Denissen MPM, Honing H (2020) Parallel pitch processing in speech and melody: A study of the interference of musical melody on lexical pitch perception in speakers of Mandarin. PLoS ONE 15(3): e0229109. The correct Funding statement is as follows: JW, MPRD and HH were supported by a Horizon grant (317-70-010) of the Netherlands Organization for Scientific Research (NWO). HH was supported by a Distinguished Lorentz fellowship granted by the Lorentz Center for the Sciences and the Netherlands Institute for Advanced Study in the Humanities and Social Sciences (NIAS).The correct Acknowledgements statement is as follows: The authors are very grateful to Loy Clements and Carlos Vaquero for their help in creating the code to generate the auditory stimuli. Additional appreciation to Loy Clements for his assistance in creating additional code for stimulus creation, EEG processing, and statistical analyses; and to Ya-Ping Hsiao and Yuan Yan for their assistance in recording and translating the Mandarin stimuli. The authors also express valued appreciation to Johan Tangerman for his help with data collection."} +{"text": "Pharmaceutical markets worldwide expose substantial stratification in their Composite Annual Growth Rates (CAGR) . This isgains in societal welfare and living standards,technological innovation in medicine,rapid urbanization in developing world regions.Successful insurance-based risk-sharing agreements made pharmaceuticals dispensing and health care services provision affordable or virtually free at the point of consumption in most OECD and many middle-income countries , 6. The There is direct historical evidence of long-term growth in pharmaceutical and overall health spending in absolute and GDP % terms worldwide , 9. The Ke et al. was to investigate the costs and benefits of an integrated care model for hypertension management in rural China using three intervention modes: multidisciplinary teams (MDT), multi-institutional pathways (MIP), and global system budget and performance-based payments (SGB-P4P). The integrated care model with performance-based prepaid payments was the most beneficial intervention in the healthcare delivery system for managing chronic diseases in China. In contrast, the general integrated care model (MDT + MIP) was not cost-effective. The integrated care model (MDT + MIP + SGB-P4P) was proposed for use in community hypertension management in rural China as a continuous, patient-centered care system to improve hypertension management efficiency .For example, hypertension has risen to become the world's second leading cause of death. However, rural China's fragmented three-level medical and healthcare system must provide continuous, coordinated, comprehensive care. As a result of the lack of total health care for hypertensive patients, rural China has a low rate of hypertension control. The purpose of the study conducted by ficiency . As a reSaxena et al.).Researchers from the Indian University, Jaipuria Institute of Management, worked on an intriguing topic involving the pharmaceutical industry's role in building resilient health systems. This study investigated the correlation between the pharmaceutical industry's current sustainability agenda, which is based on the United Nations Sustainable Development Goals (SDGs), the elements of the Joint External Evaluation (JEE) tool, and the three components of the One Health approach. This study provides insight into this area and can assist various government and non-government stakeholders in considering the integration of the pharmaceutical sector to improve health security . The study, conducted by Ye et al., aimed to compare the cost-effectiveness of Nivolumab plus ipilimumab vs. platinum plus chemotherapy for American patients' first-line treatment of MPM. The incremental healthcare costs and QALYs for Nivolumab plus Ipilimumab vs. chemotherapy were $196,604.22 and 0.53, respectively, for a total incremental cost-effectiveness ratio (ICER) of $372,414.28/QALY. The ICER for Nivolumab plus ipilimumab is higher than the theoretical willingness-to-pay threshold of $207,659/QALY in the United States, implying that first-line nivolumab plus ipilimumab for unresectable MPM may not be cost-effective compared to platinum plus chemotherapy .According to Xiang et al.).Another similar research was conducted. Camrelizumab, the first domestic programmed cell death 1 (PD-1) antibody approved for lung cancer in China, has demonstrated efficacy in patients with non-small-cell lung cancer (NSCLC). Camrelizumab combination therapy was more expensive and provided 0.11 QALYs more than chemotherapy in the base case analysis, 0.12 QALYs more in the subgroup analysis, and 0.34 QALYs more in the scenario analysis. In the base case, the ICER was $63,080 per QALY, $46,311 per QALY, and $30,591 per QALY in the subgroup and scenario analyses, respectively. The results showed that camrelizumab combination therapy is not as cost-effective as first-line therapy for NSCLC patients in China. Camrelizumab combination therapy, on the other hand, has a 62.8% chance of dropping below the WTP threshold and being cost-effective for unselected patients .An intriguing study by Luo et al. presented 18 BIA studies for anti-diabetic drugs for diabetes mellitus conducted in various countries and regions, including Europe, the United States, Asia, and South America, in their systematic review study. With the emergence of different anti-diabetic drugs, BIA is essential for determining the affordability of implementing a new anti-diabetic drug in a specific health setting.A budget impact analysis (BIA) is an economic assessment that estimates the financial implications of implementing a new intervention. BIA supplements cost-effectiveness analyses in making informed reimbursement decisions (CEAs) . Luo et This Research Topic was created to tackle the core challenges of medicines provision and medical care financing across the Globe . Its tarThe editors hope that these significant and diverse subjects will contribute to expanding existing knowledge. Furthermore, they offer an excellent opportunity to discuss the fundamental challenges of pharmaceutical care provision and healthcare financing worldwide . A diverMJ has prepared the manuscript draft. MJ, NV, KS, and KK have revised it for important intellectual content. All contributed to the article and approved the submitted version."} +{"text": "We describe the FETP - Frontline program, including implementation, structure, achievements, impact, and its role in improving the epidemiological workforce capacity of Guinea-Bissau.this cross-sectional descriptive study uses 2015-2019 program data collected through record reviews and historical narratives from FETP students and graduates. We generated descriptive summary statistics using the Guinea-Bissau's FETP-Frontline program database, student assignments, and investigation reports, after reviewing the FETP standardized curriculum and program guidelines.since its inception in 2016, FETP Frontline has implemented 14 cohorts and trained 198 frontline surveillance officers. Program participants improved surveillance data quality, investigated 51 outbreaks at national and regional levels, and contributed to disease research and surveillance in 227 separate field investigations. Participants frequently responded to priority health emergencies, including clusters or outbreaks of Zika, microencephalies, dengue, yellow fever, anthrax, malaria, and tuberculosis.Guinea-Bissau's FETP - Frontline program provides a practical example of an effective strategy to strengthen health systems through a well-prepared workforce trained to quickly detect and respond to health threats. Guinea-Bissau is one of the smallest countries in West Africa, with an estimated population of 1.8 million inhabitants, divided into 8 administrative regions . The coucentral level, there is the Ministry of Public Health (MINSAP) and the National Institute of Public Health (INASA). The INASA was established in 2011 as a government agency that has authority over epidemiology, surveillance, health communication, public health laboratory system, and health research. The intermediate level consists of 11 health regions, including the Autonomous Sector of Bissau (SAB). Health regions offer technical support and coordination of health areas that constitute the peripheral level [The national health system comprises three levels of care: central, intermediate, and peripheral. At the al level . Each ofGuinea-Bissau uses the Integrated Disease Surveillance and Response (IDSR) strategy to detect priority diseases, conditions and events, and report across all health system levels in the country . This inIn January 2015, the US Centers for Disease Control and Prevention (CDC) and the Global Network of Training Programs in Field Epidemiology and Public Health Interventions (TEPHINET) implemented an emergency training strategy - Surveillance Training for Ebola Preparedness (STEP) - to rapidly increase surveillance capacity in health regions and along areas bordering affected countries ,10. The This paper aims to describe the implementation of FETP-Frontline in Guinea-Bissau, its development and organization, the major outputs of the program, and its contribution to the country's public health system. Our main hypothesis was that FETP-Frontline will contribute to a stronger public health system through improved disease surveillance, data collection, analysis, reporting, and frontline response.2 to visualize the geographic distribution of trained FETP-Frontline graduates. We also reviewed the program guidelines to describe the planning, curriculum, and implementation process. FETP-Frontline participants were selected based on recommendations from INASA and other health institutions, availability, interest, and minimum skillset to complete the program.This descriptive study documents the history and functionality of Guinea-Bissau's FETP-Frontline between 2015 and 2019. For secondary data analysis, we accessed the program's student and alumni database, program guidelines, classroom and field assignments, and investigation reports completed during the implementation of the program. We used to Microsoft Excel for data management and to perform summary statistics of participant profiles between 2016-2019, including the number of cohorts, graduates by profession, and activities performed. We used QGISFunding: the funding for the Guinea-Bissau FETP was provided by the United States Centers for Disease Control and Prevention (US CDC) through a cooperative agreement with AFENET number CDC-RFA-GH15-1619 for Strengthening Applied Epidemiology and Sustainable International Public Health Capacity through Field Epidemiology Training Programs.Disclaimer: the findings and conclusions in this manuscript are those of the author(s) and do not necessarily reflect the official position of the Centers for Disease Control and Prevention.Field Epidemiology Training Programs (FETP)are 3-tiered in-service training programs for public health professionals to strengthen their field epidemiology and surveillance skills. The goal of FETP is to build field epidemiology capacity at all levels of a country's health system, starting from the district (FETP-Frontline) to the regional (FETP-Intermediate) and the national (FETP-Advanced) levels.The FETP-Frontline in Guinea-Bissau aimed to develop and strengthen the epidemiological capacity in the country to strengthen public health surveillance and evidence-based decision-making, particularly at the local level of the surveillance system. The program is coordinated by the INASA and receives technical and administrative assistance from CDC-Atlanta and the African Field Epidemiology Network (AFENET), respectively.Program selection: the FETP-Frontline integrated training for human and animal health professionals to enhance global health security from different and complementary viewpoints [ewpoints ,11. The Guinea-Bissau's FETP-Frontline Program: Guinea-Bissau's FETP-Frontline used the CDC standardized curriculum translated to Portuguese and adapted to Guinea-Bissau's training needs [ng needs . The thrstandardized curriculum for FETP-Frontline is divided into 3 classroom workshops and 2 fieldwork blocks in between. The program provided additional workshops in GIS and Epi-Info. During the training process, the participants learned and practiced fundamental epidemiological skills used in surveillance including disease detection, reporting, and case investigation and response at the local level. They used case definitions, simple tables, and graphs to apply their skills to summarize and present epidemiological data.The Workshop 1 was held in 5 days, where we covered surveillance data collection, analysis, and interpretation, data quality, and basic principles of biostatistics. After this workshop, the professionals returned to their field of work, usually related to epidemiological surveillance. During the 4 weeks of fieldwork, the students conducted a data quality audit and a surveillance data analysis summary report.Workshop 2 also lasted 5 days. It was an opportunity for participants to present their data analysis and delve deeper into surveillance questions and discussions. They were also introduced to the basic concepts of scientific communication and case/outbreak investigation and response. After the second workshop, residents returned to their workplace, where they conducted field projects to practice, implement, and reinforce what they learned. This included participating in outbreak investigations and/or conducting case investigations, while ensuring thorough analysis of surveillance data.workshop 3, where the professionals presented their results to FETP colleagues, country's health managers, stakeholders, and local health authorities. Graduating participants received a course completion certificate. At the end of the program, each participant submitted the following products: (1) Data quality evaluation report with recommendations for improvement; (2) Surveillance summary report; (3) Outbreak investigation report; (4) Oral presentation of the surveillance summary.The course finalized at The classroom sessions were conducted by epidemiologists and public health professionals from different health agencies, such as Guinea-Bissau's FETP-Frontline graduates, CDC Atlanta, World Health Organization (WHO), INASA, National Public Health Laboratory, Portuguese Cooperation, Bandim Health Project, and M\u00e9decins Sans Fronti\u00e8re (MSF). During the program, each participant was assigned a mentor who was in regular contact with them and provided feedback and guidance, as needed, for the successful completion of the field project. The ratio of mentors to mentees was 1: 3.Pre and post-test questionnaires, completed by trainees at the beginning of the workshops and upon return from the field, were used to evaluate the course. Evaluation questions were related to the content covered in each of the workshops and the practical activities. Results from the questionnaires were presented during the final workshop, where residents had an opportunity to discuss and request clarifications. For each cohort, we completed a post-training evaluation to assess the impact of the course on the graduates' work routine. A strategy is being developed for this type of evaluation to be incorporated into the standardized program curriculum.FETP-Frontline implementation: in November 2016, CDC completed an in-country assessment to implement the FETP program in INASA. In addition, a resident advisor was hired to support program implementation. In March 2016, FETP-Frontline was officially launched. In 2017, the program expanded, adding 2 regional and 2 national cohorts. From 2016 to 2019, Guinea-Bissau's FETP-Frontline implemented 14 cohorts and successfully trained 198 graduates. The program trained professionals from the national or regional levels (6 cohorts) and from the local level (8 cohorts) of the health system. All graduates conducted a descriptive study with surveillance data and 83 (42%) participated in at least one outbreak investigation or emergency response in public health within the framework of the Lusophone FETP Network. The main objective is to strengthen Guinea-Bissau's FETP and the overall public health system, by training Guinean professionals in Brazil's FETP Advanced 2-year program. The first medical professional who completed the course in Brazil returned to Guinea-Bissau, where he assumes a surveillance management post. In 2020, two additional professionals completed Brazil's FETP Advanced training, and three other Guinean residents graduated from the West Africa Regional Advanced FETP Program, located in Burkina Faso. Guinea-Bissau's FETP-Frontline has been an AFENET and TEPHINET member since 2017, where they can participate in conferences, receive technical support and funding, and network with other programs.In 2016, the FETP-Frontline in-service training strategy was successfully implemented in Guinea-Bissau and supported the country's effort to respond swiftly to priority public health events. This paper describes the implementation of the program, its development, organization, main outputs, and its contribution to the country's public health system from 2016-2019. During this time period, the program successfully trained 198 graduates in FETP-Frontline where each participant conducted a descriptive surveillance project and 42% assisted in outbreak investigations or emergency public health responses. Globally, FETP-Frontline has proven to be a practical solution to improve disease surveillance and response in low- and middle-income countries . Guinea-The main challenge with implementing Guinea-Bissau's FETP-Frontline is guaranteeing the program's sustainability. The program receives financial support, transited from CDC funds to the World Bank, through the Regional Disease Surveillance Systems Enhancement (REDISSE). However, the goal is to build public health capacities independent to foreign aid and support. The country may consider how the FETP can be incorporated and sustained within the country's current health system. Therefore, a steering committee was established to discuss how to strengthen the program and establish it into the national surveillance network.Second, Guinea-Bissau has limited laboratory diagnostic capacity, which the government is working hard to improve. Some disease samples must be sent to the Institute Pasteur in Dakar, Senegal, for diagnostic confirmation. Laboratory diagnosis is a key component in outbreak investigations, and delays in receiving results from reference laboratories limits the interpretation of epidemiological data.Third, few graduates have the availability, financial resources, and adequate technical preparation to become mentors and continue to support the program and One Health approach to field epidemiology. Out of the 198 Frontline graduates, only 9 worked in laboratory settings, 5 worked in the animal health sector, 2 in the social sciences. Identifying and maintaining highly qualified and committed mentors is still a major concern among FETPs globally ,17. SimiFourth, the technical and operational capacity of human resources is limited at the national and sub-national levels. Retaining graduates within the health system is key for the program's long-term objectives of strengthening the health system, including successful mentorship.Finally, one of the principles of epidemiology in public health is information sharing of relevant study findings through reports, publications, and other means of scientific communication. Given that trainees of Guinea-Bissau's FETP-Frontline program are not native English speakers, they face challenges in publishing their field work with the broader public health community. To overcome this obstacle, a Scientific Writing Program and Calibrated Peer Review exercises, can help improve scientific writing skills for residents to showcase their research in the international arena ,19.The national limitations of the capacity to respond to outbreaks, perform laboratory investigations, and implement fieldwork recommendations have prevented the progress of some studies. However, the program's benefits are unquestionable, as they strengthen and build epidemiological capacity and public health surveillance. Guinea-Bissau's pyramid model of field epidemiology training is still underdeveloped, but is expected to implement the next tier, FETP Intermediate, by 2024.FETP-Frontline in Guinea-Bissau is an effective strategy to respond to public health emergencies and improve a country's local surveillance system, through in-service training of health professionals involved in data generation, analysis, scientific communication, and evidence-based recommendations for decision making. However, to sustain the FETP-Frontline program in the long term, it is essential for the country to have a structured programmatic approach that considers the program's financial, operational, and technical sustainability.The purpose of FETP-Frontline is to strengthen a country's surveillance system at the local level;Committed and experienced mentors, as well as selecting suitable trainees, are important to ensure the quality and impact of the program.Guinea-Bissau's FETP-Frontline experience confirms the importance of strengthening surveillance at the local level;The FETP-Frontline training course has improved the country's ability to rapidly detect and respond to health emergencies in Guinea-Bissau."} +{"text": "J. Exp. Biol. (2022) 225, jeb243766 (doi:10.1242/jeb.243766).There was an error in 0000-0003-2902-7568.The ORCID iD for Alice Lowry should read: Additionally, in the first sentence of Materials and Methods, the number of harbour seals is incorrectly given as \u201835 males, 33 females\u2019; it should instead be \u201834 males, 34 females\u2019. The number of males and females is correctly reported in paragraph two of Materials and Methods, Table S1 and the dataset, and does not affect the results in the article or the conclusions of this study.Both the online full-text and PDF versions of the paper have been corrected. The authors apologise for these errors and any inconvenience they may have caused."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "BC data were used as the gold standard. Race and ethnicity were categorized as non-Hispanic (NH)-White, NH-Black, Hispanic, NH-Asian Pacific Islander (API), and NH-American Indian or Alaskan Native (AIAN). Overall, race and ethnicity misclassification and its impact on SMM at the state- and hospital levels were estimated. At the state level, NH-AIAN women were the most misclassified : 25.2%) and were commonly classified as NH-API (30.3%) in HD records. NH-API women were the next most misclassified and were commonly classified as NH-White (5.8%) or NH-other (5.5%). At the hospital level, wide variation in sensitivity and PPV with negative skewing was identified, particularly for NH-White, Hispanic, and NH-API women. Misclassification did not result in large differences in SMM rates at the state level for all race and ethnicity categories except for NH-AIAN women (% difference 78.7). However, at the hospital level, Hispanic women had wide variability of a percent difference in SMM rates and were more likely to have underestimated SMM rates. Reducing race and ethnicity misclassification on HD records is key in assessing and addressing SMM differences and better informing surveillance, research, and quality improvement efforts.Hospital discharge (HD) records contain important information that is used in public health and health care sectors. It is becoming increasingly common to rely mostly or exclusively on HD data to assess and monitor severe maternal morbidity (SMM) overall and by sociodemographic characteristics, including race and ethnicity. Limited studies have validated race and ethnicity in HD or provided estimates on the impact of assessing health differences in maternity populations. This study aims to determine the differences in race and ethnicity reporting between HD and birth certificate (BC) data for maternity hospitals in Florida and to estimate the impact of race and ethnicity misclassification on state- and hospital-specific SMM rates. We conducted a population-based retrospective study of live births using linked BC and HD records from 2016 to 2019 ( Severe maternal morbidity (SMM) is an unexpected life-threatening complication of childbirth in which women suffer short-term or long-term health consequences if not identified and treated in a timely manner . In the SMM mostly affects women who identify as racial and ethnic minorities ,6, with The study objectives were as follows: (1) determining the differences in race and ethnicity reporting between HD and birth certificate (BC) data from maternity hospitals in Florida and (2) estimating the impact of race and ethnicity misclassification on state- and hospital-specific SMM rates.We used the latest available data to conduct a population-based retrospective study using Florida BC linked to inpatient HD records for live births from January 2016 to December 2019. The study time period was chosen because 2016 was the first full year of HD data following the ICD-10-CM implementation. BC records were provided by the Florida Department of Health and contain demographic and clinical information on the mother and baby. HD records were provided by the Florida Agency for Health Care Administration and contain individual-level patient data for each inpatient hospital stay. Hospitals are required to submit race and ethnicity in a standard format to AHCA. However, how this information is actually collected varies across hospitals and may be self-reported by patients or designated from observation by admitting clerks or clinical staff ,19,20,21n = 42,931). We also excluded births with mothers reporting multiracial heritage on the BC because multiracial heritage is not captured on the HD record. The study population at the state level included 727,079 births from 106 hospitals. Hospital-level and hospital-level SMM populations have further exclusions and are described later.This study analysis consisted of three different subgroups, henceforth referred to as the state-level, hospital-level, and hospital-level SMM populations . The oriWe extracted race and ethnicity information from both the BC and HD data sources. The HD record allowed the classification of race into categories . Due to Since mothers\u2019 self-reported race and ethnicity as documented on the BC, are typically based on US recommendations , the BC We calculated the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) separately for (1) race, (2) ethnicity, and (3) combined race and ethnicity variables to analyze state-level misclassification between the BC and HD using state-level population. Sensitivity estimates the probability that the HD captures race and ethnicity as reported in the BC. For example, we calculated the sensitivity of NH-AIAN as the proportion of all individuals reported as NH-AIAN on the BC that were correctly identified as NH-AIAN on the HD. PPV estimates how frequently the HD was accurate in classifying a specific race and ethnicity among those who reported as that race and ethnicity. For example, we calculated the PPV for NH-AIAN as the proportion of NH-AIAN on the HD that was classified as NH-AIAN on the BC. We made these calculations in two ways. First, we considered missing race and/or ethnicity in one data source and non-missing race and/or ethnicity in the other source as disagreement. Second, we excluded records with missing race and ethnicity and estiWe then assessed the impact of maternal race and ethnicity misclassification on SMM at the state level by first calculating the number of SMM cases, SMM rates, and risk ratios representing the association between race and ethnicity and SMM using each data source individually. We then calculated the percent difference in these SMM statistics when using race and ethnicity from the HD versus the BC data.To further examine misclassification at the hospital level, we used a subset of the state-level population. The hospital-level population excluded nine additional hospitals that did not have at least 20 births from each race and ethnic group\u2014NH-White, NH-Black, Hispanic, and NH-API\u2014in order to ensure statistically reliable estimates for each race and ethnicity category when calculated at the hospital level. The hospital-level population consisted of 706,023 births from 97 hospitals. We analyzed the distribution of sensitivity, specificity, PPV, and NPV measures across hospitals. To then assess how race and ethnicity misclassification affects SMM rate at the hospital level, we used a subset population of the hospital-level population, where we excluded an additional 58 hospitals that did not have at least 10 births with SMM from each of the following groups: NH-White, NH-Black, and Hispanic. Again, this was performed in order to have minimally reliable estimates of SMM rates at the hospital level; insufficient numbers of NH-API were available at the hospital level for inclusion. The hospital-level SMM population consisted of 428,718 births from 39 hospitals. We conducted a sensitivity analysis comparing the sensitivity and PPV findings for these 39 maternity hospitals to the overall 97 maternity hospitals in the state. For each race/ethnic group, we then examined the distribution of hospital-level percent differences in rates of SMM diagnosed in maternal delivery HD records compared to BCs.All analyses for this study were conducted using SAS software version 9.4 , and figures were generated using R version 4.1.3 with RStudio 2022.02.3.Race and Ethnicity. At the state level, NH-AIAN was the most misclassified race and ethnicity and was commonly misclassified as NH-API (30.3%) in HD records (see ords see . NH-AIANRace and Ethnicity Separately. The sensitivity and PPV for each race group were fairly similar to the findings of the combined category (see gory see . As a seAt the hospital level, wide variation in sensitivity and PPV with negative skewing was identified, particularly for NH-White, Hispanic, and NH-API groups . The vioGenerally, misclassification did not result in large SMM rate differences at the state level for most race and ethnicity categories . HoweverWe found that misclassification at the state level varied amongst all racial and ethnic groups, i.e., NH-White, NH-Black, Hispanic, NH-API, and NH-AIAN groups. The most misclassified groups were NH-API and NH-AIAN women, and the least misclassified group was NH-Black women. A wide variation in misclassification for all races and ethnic groups was observed at the hospital level. Misclassification of race and ethnic groups resulted in differences in SMM rates for the NH-AIAN group at the state level and among all three groups at the hospital level, especially the Hispanic group.At the state level, Florida\u2019s sensitivity and PPV of HD data for race and ethnicity classification at delivery varies widely across subgroups. NH-API and NH-AIAN women were by far the most misclassified group; NH-Black women were the least misclassified. Howland et al. is the oRace and ethnicity misclassification did not result in major differences in SMM rates across NH-White, NH-Black, Hispanic, and NH-API women and is consistent with findings by Howland et al. . HoweverAt a hospital level, wide variation in sensitivity and PPV exists, resulting in frequently unreliable hospital data for race and ethnicity groups. Most hospitals accurately classified NH-Black and NH-White women, with a sensitivity of at least 85%. However, 25% and 50% of hospitals did not capture race and ethnicity correctly for Hispanics and NH-API, respectively. While classification at the state level for NH-White, NH-Black, and Hispanics was mostly accurate, the extent of race and ethnic misclassification across hospitals is inconsistent, making it challenging to predict the impact on outcome rates for a given hospital.Wide accuracy variation in hospital sensitivity and PPV can be misleading and result in considerable underestimation of SMM rates for some race and ethnic groups, especially Hispanics. Despite our ability to assess the impact of NH-API and NH-AIAN misclassification on SMM rates at a state level, we anticipate that these two groups would be more greatly impacted at a hospital level. Our findings provide one of the first looks at the hospital level on the prevalence of racial and ethnic misclassification among maternity patients overall and the impact on maternal outcomes.Our study demonstrated wide variability of race and ethnicity misclassification in HD records at the state level for all race and ethnic groups, especially for the NH-API and NH-AIAN groups, and wide variability at the hospital level for NH-White, Hispanic, and NH-API groups. Misclassification at the state level did not result in large SMM rate differences, except for the NH-AIAN group, where we observed an overestimation of SMM rates. At the hospital level, misclassification appeared to result in an underestimation of SMM rates for all three groups but especially the Hispanic group. This misclassification likely impacts multiple health and health quality measures at a hospital level. Therefore, health agencies and hospitals should not rely on state-level race and ethnicity data as a proxy for assessing accuracy at the hospital level. This study highlights the need for uniform processes in collecting self-reported race and ethnicity, nativity, and country of origin, and patient education on the importance of ascertaining this information in order to improve their health care. Importantly, correct classification of race and ethnicity would potentially ensure that the delivery of health care services is tailored more appropriately.Health differences stemming from race and ethnicity are a health priority because they can be used to assess unmeasured social and health factors that influence health and health care delivery. To obtain a true measure of outcome differences, data sources used to identify race and ethnicity must be accurate and reliable. In the US, reducing race and ethnicity misclassification in HD data is key to monitoring and reducing SMM and other maternal health outcomes at the state- and hospital level, where improvements can be made. Future studies are needed to further delineate the implications of misclassification among people of multiracial heritage on health outcomes at a state and hospital level.One strength of the study is Florida\u2019s large, highly diverse birthing population, which provides a large sample size for population subgroups. This enabled assessing smaller race and ethnicity categories such as NH-AIAN and NH-API. Second, it has become a priority to obtain accurate estimates of race and ethnicity data at the state-, county-, and hospital levels because of the increasing diversity across many US states and in an effort to monitor and address disparities . TherefoThis study assessed the differences in race and ethnicity reporting between HD and BC data from maternity hospitals in Florida and estimated the impact of race and ethnicity misclassification on state- and hospital-specific SMM rates. Misclassification of race and ethnicity in HD records occurs at both the state and hospital levels, resulting in differences in SMM rates at each level. The role of race and ethnicity may be different in other countries. Reducing race and ethnicity misclassification is key in assessing and addressing SMM rate differences and better informing surveillance, research, and quality improvement efforts."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.1As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.3Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.5Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.7Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.16Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.22Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.10In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.1As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.3Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.5Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.7Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.16Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.22Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.10In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.1As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.3Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.5Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.7Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.16Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.22Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.10In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Background: Drug manufacturing and distribution is a complex, global process. The global drug supply chain is prone to disruptions associated with geopolitical issues, trade, civil unrest, severe weather, and pandemics, all of which have the potential to affect medication supply and result in drug shortages. To our knowledge, the extent to which the supply of antimicrobials is threated due to disruptions in the drug supply chain in the United States is unknown. We examined trends and duration of disruptions to the drug supply chain for antimicrobials. Methods: Manufacturer reports of supply disruptions were extracted from the Food and Drug Administration (FDA) and the American Society for Health-Systems Pharmacists (ASHP) websites and merged on the agent-formulation level. For each month of the study period, a drug was considered to have an active supply chain issue if an FDA or ASHP shortage or recall report overlapped with that month for \u226515 days, or if a discontinuation had occurred within the previous 3 months. Total months of supply chain issues were summed for antimicrobials overall, at the agent formulation , and class levels. A Mann-Kendall test was used to determine the significance of trends in supply-chain issues. Results: Of 105 antimicrobials purchased in the United States, 74 (70%) had a supply-chain issue for \u22651 month from January 15, 2017, to June 30, 2022. Combined, the 74 agents had 1,611 total months of supply-chain issues over the 66-month study period. Agents from the penicillin class were most frequently affected , but cephalosporins had supply-chain issues for the longest duration . From 2017\u20132021, supply-chain issues decreased significantly for penicillins and quinolones . No trend was identified for the other classes or antimicrobials overall. Interestingly, supply-chain issues for most classes did not increase with seasonal increases in antimicrobial use. Also, supply-chain issues affected 33 antimicrobial agents for at least half of the study period, and supply-chain issues affected ampicillin-sulbactam, cefotaxime, ceftazidime, cefotetan, cefepime, clindamycin, vancomycin for 100% of the study period. Conclusions: Drug supply-chain issues commonly affect antimicrobials and are not improving for most classes. Drug supply-chain issues cause significant strain on healthcare, including drug procurement, access to optimal therapy, and poses challenges to prescribing and antimicrobial stewardship. To decrease the threat to the antibacterial drug supply, action should be taken to strengthen the drug supply chain to ensure access to these essential medicines.Disclosures: None"} +{"text": "In this number the Int Braz J Urol presents original contributions with a lot of interesting papers in different fields: Robotic Surgery, Prostate Cancer, Endometriosis, Translational Research, Male Health and Renal stones, Kidney Cancer, Bladder Cancer and UPJ obstruction. The papers came from many different countries such as Brazil, Italy, China, Saudi Arab and USA, and as usual the editor\u00b4s comment highlights some of them. The editor in chief would like to highlight the following works:The May-June number of Int Braz J Urol is the 22Dr. Wang and collegues from China, presented in page 281 (Dr. Ribeiro-Julio and collegues from Brazil, presented in page 299 (Dr. da Silva and collegues from Brazil, performed in page 320 (Dr. Moratti Gilberto and collegues from Brazil performed in page 334 (Dr. Raed and collegues from Saudi Arab permormed in page 372 a nice sDr. Sobhani and collegues from USA permormed in page 351 a nice sDr. Lv and collegues from China performed in page 359 the coveThe Editor-in-chief expects everyone to enjoy reading."} +{"text": "E. coli and K. pneumoniae species. This study was aimed at analyzing the state of hospital-acquired, carbapenem-resistant E. coli and K. pneumoniae in the United Kingdom between 2009 and 2021. Moreover, the study analyzed the most efficacious approaches to patient management for controlling the carbapenem-resistant Enterobacteriaceae (CRE) spread. Initially, 1094 articles were identified as relevant for screening, and among them, 49 papers were eligible for full-text screening, with a total of 14 articles meeting the inclusion criteria. The information was recorded from published articles through PubMed, the Web of Science, Scopus, Science Direct, and the Cochrane library and was used to search for hospital-acquired carbapenem-resistant E. coli and K pneumoniae in the UK between 2009 and 2021, in order to evaluate the spread of CRE in hospitals. The total number of carbapenem-resistant E. coli was 1083 and this was 2053 for carbapenem-resistant K. pneumoniae in more than 63 UK hospitals. KPC was the dominant carbapenemase produced by K. pneumoniae. The results showed that the treatment options considered depended on the type of carbapenemase produced; K. pneumoniae showed more resistance to a treatment options, i.e., Colistin, than the other carbapenemase. The current state of the UK is at minimal risk for a CRE outbreak; however, appropriate treatment and infection control measures are highly required to prevent this CRE spread at the regional and global levels. The present study findings have an important message for physicians, healthcare workers, and policymakers about hospital-acquired carbapenem-resistant E. coli and K. pneumoniae spread and approaches to patient management.Worldwide, hospital-acquired infections (HAIs) are continuously rising within healthcare settings, leading to high mortality and morbidity rates. Many hospitals have reported the spread of carbapenemases globally, specifically within the E. coli and Klebsiella spp. (both are species within the Enterobacteriaceae family), and these infections are at their peak during the summer in North America, the Middle East, Europe, Asia, and Australia [A hospital-acquired infection (HAI) is an infection that is acquired in a healthcare setting within two days of admission . The thrustralia . The majustralia .Klebsiella pneumonia carbapenemase (KPC), New Delhi metallo-\u03b2-lactamase (NDM), and oxacillinase (OXA), which hydrolyze carbapenem [K. pneumoniae and E. coli species [Recently, Enterobacteriaceae species were shown to be highly resistant to broad-spectrum antibiotics through extended-spectrum \u03b2-lactamases (ESBLs), due to the acquisition of gene-encoding enzymes, which upregulate efflux pumps and alterations to the antibiotic binding sites . Thereforbapenem ,8. These species .K. pneumoniae has been reported as the leading cause of bacteremia and pneumonia in the United Kingdom (UK) [K. pneumoniae (CRKP) is a major health-related pathogen responsible for pneumonia, surgical site infection, bloodstream infection, UTIs, and meningitis, leading to high morbidity and mortality rates [E. coli and Klebsiella are the leading causes of hospital-acquired infections worldwide [E. coli became increasingly resistant to \u03b2-lactams and the carriage of resistant AmpC plasmids [E. coli infections. However, carbapenem-resistant E. coli started to emerge, mainly through the production of carbapenemases, along with a decreased antibiotic permeability (increased efflux pump and lack of porin presence) and an alternation of the carbapenem binding sites [dom (UK) . Currentty rates ,12. E. corldwide . In prevplasmids . Carbapeng sites ,16.K. pneumoniae (CRKP) and CRE E. coli, a detailed examination of the spread of CRE strains in the UK in the past decade appears to be limited in the literature. Therefore, this study investigates the spread of CRKP and CRE E. coli in the UK from 2009 to 2021. Additionally, it identifies the most common type of carbapenemases carried by CRKP and CRE E. coli and the antibiotic treatments used, depending on the type of carried carbapenemase.Despite the wide spread of CRE strains, specifically carbapenem-resistant The present data-based observational study was conducted under the College of Biomedical Science, University of Bristol, Bristol, United Kingdom and School of Life Sciences, Faculty of Health & Life Sciences, Coventry University, Coventry University, United Kingdom and College of Medicine, King Saud University, Riyadh, Saudi Arabia during the period from January to October 2022.For the search of the literature, the Preferred Reporting Items for Systematic and Meta-Analysis (PRISMA) guidelines were followed while conducting this study, in order to select the reliable sources that correlated with the research aims. The protocol was registered on PROSPERO (registration number CRD42022303741).E. coli and K. pneumoniae to investigate the most effective methods for managing HA-CRE. Cases of CRE in the UK published in scientific journals between 2009 and 2021 were used to evaluate the spread of CRE in hospitals. Many studies started HA-CRE data collection in UK hospitals in 2009; thus, 2009 was set as the starting line of this study. The minimum inhibitory concentration (MIC) antibiotic treatment options were analyzed to identify the resistant and susceptible strains, depending on the type of carbapenemase, NDM, KPC, OXA, VIM, and IMP produced by the reported CRE E. coli and K. pneumoniae.In this study, data were retrieved about the incidence and prevalence of hospital-acquired carbapenem-resistant E. coli and K. pneumoniae within the CRE species. In contrast, studies performed on non-UK reported cases, data reported before 2009, carbapenem-susceptible Enterobacteriaceae, data that did not specify the Enterobacteriaceae species, and community-acquired infections were all excluded. Moreover, brief communications, editorials, and review articles were also excluded.The primary sources of the reported carbapenem cases were collected using scientific search engines. The inclusion and exclusion criteria determined the papers that were included or rejected for the review and data collection. The included studies were those published in the UK that contained reported cases, quantitative records of CRE-infected patients, and E. coli and/or K. pneumoniae during >2 days of a hospital stay in the UK between 2009 and 2021. The literature was searched using scientific research engines, including PubMed, the Web of Science, Science Direct, Scopus, and the Cochrane Library to find the relevant articles. The search terms used were the Boolean operators \u2018AND\u2019, \u2018OR\u2019, and \u2018NOT\u2019 and included \u2018carbapenem-resistant Enterobacteriaceae\u2019 OR \u2018Carbapenem-resistant Escherichia coli\u2019 OR \u2018Carbapenem-resistant Klebsiella pneumoniae\u2019 AND \u2018Hospital-acquired infection\u2019 OR \u2018nosocomial infection\u2019 OR \u2018healthcare-acquired infection\u2019 AND \u2018United Kingdom\u2019 OR \u2018England\u2019 OR \u2018Wales\u2019 OR \u2018Northern Ireland\u2019 OR \u2018Scotland\u2019. Furthermore, the scope of the search was expanded by screening the reference list of the relevant articles to identify topic-related articles. Only full-text articles published in the English language were included, and, following the removal of duplicated articles, the selection of the papers was based on three sections: (1) titles, (2) abstracts, and (3) results and full-text screening.This study analyzed the published literature and research articles on reported patients who acquired carbapenem-resistant E. coli and K. pneumoniae, the type of carbapenemase, the sample type , and the age group. However, the articles that presented MIC values from patient-identified CRE isolates were separately recorded using the Excel sheet, followed by a total summary of all the values from the correlated papers for all the antibiotics. For each antibiotic, the susceptibility, intermediate, and resistance breakpoint values were determined using the European Society of Clinical Microbiology and Infectious Diseases (EUCAST) breakpoint tables.At the post-inclusion and exclusion stage, Excel and EndNote were used to uniquely select the relevant data and identify which factors were presented by each article. Each article was presented in a row on an Excel sheet to represent the information provided by the article. The columns presented the total number of CRE reported cases, the hospital location, the specific quantity of carbapenem-resistant E. coli and K. pneumoniae using Statistics Package Social Science (SPSS) Version 25 . Statistical results with a p-value below 0.05 were considered as significant. Additionally, the percentage, frequency, mean, and median were calculated. The data conducted are variables of numerical data, which are non-continuous discrete values. A chi-squared (\u03c72) analysis was performed, using SPSS Version 25, with the following formulae: p-value, which was necessary for determining whether to reject the null hypothesis or not. In this study, data were retrieved from the publicly available literature, hence ethical approval was not required.The statistical analyses were performed separately for both During the literature search, 1094 retrieved articles were identified as relevant for screening, and among them, 49 papers were eligible for full-text screening, with a total of 14 articles ,28,29,30E. coli infections , while OXA-48-Like plasmid in E. coli (94.3%) was much higher, compared to only 5.7% in CRKP.The chi-square test for the observed data yielded a result of 1403.91 with two degrees of freedom, and the and CRKP . The perE. coli, specifically KPC, NDM, and OXA-48 carbapenemases and the type of antibiotic treatments that were used to treat the infected patients. This study specifically focused on the two Enterobacteriaceae species that are most likely to spread within hospitals worldwide, CRKP and CRE. E. coli, with a detailed analysis of fourteen different articles. Different specimens were taken, including blood, urine, and sputum, from hospitalized patients, which showed that CRKP was approximately twice as prevalent as CRE E. coli. According to the European Centre for Disease Prevention and Control, both CRKP and CRE E. coli are detected by less than 1% in the UK and their spread is regional [E. coli in the UK, there are still fewer than one hundred cases per year [K. pneumoniae is less susceptible to antimicrobials compared to the other plasmids carried by K. pneumoniae. It is thought that the infection control measures put in place, such as strict patient cohorts, the correct choice of treatment, and only using the antimicrobial peptide colistin as a last resort treatment option, are the reasons that the spread of such a resistant strain has been controlled thus far [This study aimed at presenting the state of CRKP and CRE regional . Howeverregional . Additioregional . Althougper year . The sprthus far .E. coli cases were found in Manchester. The city of Manchester showed an estimated 10\u00d7 higher detection rate of CRE compared to other areas of England [It was observed that high percentages of both CRKP and CRE England . Similar England .K. pneumoniae is the most commonly reported isolate that produces carbapenemases through plasmid uptake, specifically KPC-carrying plasmids [K. pneumoniae was recorded in the UK between 2008 and 2010 [Depending on the type of carbapenemase produced by Enterobacteriaceae, the treatment and management options differ. It was observed from the studies that the NDM, IMP, and VIM carbapenemases are minority groups, being less than 1% of the total reported HA-CRE. Public Health of England reported more than 100 NDM-producing CRE isolates between 2012 and 2015 . The reaplasmids . Only onand 2010 . The outand 2010 .E. coli. However, both species showed a high percentage of susceptibility to tigecycline, colistin, and gentamicin. OXA-48-like remains susceptible to many antibiotics, including broad-spectrum cephalosporins [E. coli and K. pneumoniae.Six out of the fourteen collated studies investigated the MICs of antibiotics in CRE-infected patients. The treatment options using antibiotics were examined for more than 500 patients. Although antibiotic treatment options are very limited against MDR bacteria, it was observed that some CRE were susceptible to more than one antibiotic. Antibiotics such as tigecycline, aminoglycosides, and the antimicrobial peptide colistin were described as the optimal treatments for MDR Gram-negative infections . Due to osporins . Moreoveosporins . As obseE. coli in the UK, there is still a limited number of cases reported, which is less than 100 cases per annum [Although there has been an upsurge in the rate of carbapenem-producing er annum . It is ber annum . Howeverer annum . MoreoveTo limit the spread of threatening CRE infections, 39 European countries, including the UK, agreed to minimize the burden of CRE infection by keeping recorded CRE-positive patient data between the European Centre for Disease Prevention (ECDP) and Control in Stockholm . PreviouE. coli and CRKP, with the identification of carbapenemase, KPC, NDM, and OXA-48. This study included data from more than 60 hospitals around the UK since 2009, which made this study more specific for the identification of areas with high levels of infection. Furthermore, a selective identification of the most common carbapenemase in each species provided an overview of the most common and most resistant types of carbapenemase.To the authors\u2019 knowledge, this is the first comprehensive study which analyzed the published data for both CRE E. coli. Additionally, very limited data were available regarding the IMP, VIM, and NDM carbapenemases. Furthermore, the majority of the articles were published based on the data from hospitals in England, and the extent of the CRE spread in hospitals in Scotland, Wales, and Northern Ireland is currently unclear. Therefore, a future investigation into the CRE spread in these geographical areas is required.Its limitations, by contrast, include the fact that a few articles were CRKP-specific and did not investigate the presence of HA-CRE E. coli and CRKP situation from more than 60 hospitals and identified 11 different antibiotics that could be used for the treatment of CRE, thus providing a summary of the potential treatment options, depending on the type of carbapenemases carried. The study\u2019s findings reveal a low risk of CRE outbreak; however, appropriate treatment, alongside appropriate infection control measures, is highly required to prevent this CRE spread. The study\u2019s findings can be used to inform the IPC measures taken by the UK government to prevent future outbreaks. However, it is worth noting that further studies may be required to confirm these findings and ensure the effectiveness of the identified treatment options. Moreover, such studies should be conducted worldwide, including in the USA, Europe, Asia, and the Middle East, to understand and prevent the spread of CRE infection.This study provided the current CRE"} +{"text": "Funding statement. The Funding statement for the State Key Laboratory of Arid Land Crop Science, Gansu Agricultural University, China was displayed as \u201cNo. GSCS-20202-08\u201d. The correct Funding statement appears below:In the published article, there was an error in the \u201cThis study was supported by the research program sponsored by the State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, China (No. GSCS-2020-08).\u2019\u2019The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "At present, the field of the Internet of Things (IoT) is one of the fastest-growing areas in terms of Artificial Intelligence (AI) and Machine Learning (ML) techniques. The utilization of AI and ML is increasingly intertwined with IoT. AI, ML, and deep learning are now being utilized to make IoT services and devices smarter and more secure. Developments in IoT are playing a significant role in our daily lives. In IoT, a large number of devices such as actuators and sensors are being deployed and connected for the collection of different types of data, such as healthcare, transportation, public safety, energy, manufacturing, and smart city infrastructure espousing systems. ML/DL has also shown substantial success in the transformation of complex and massive datasets into precise comprehension as output, which can significantly facilitate intelligence, analysis, automation, and decision-making. ML has provided a way to perform giant modeling and intelligence with the integration of developments in big-data analytics, big-networking technologies, and big-data computing to achieve enormous accomplishments in diverse areas. Sensors aims to report on some of the recent innovative studies on advanced techniques in artificial intelligence and machine learning. In [The Special Issue of ning. In , the autning. In , the autning. In . The K-mIn , a ConvoIn , the autIn , the autIn , the aut"} +{"text": "Nature Cancer 10.1038/s43018-023-00630-y, published online 21 September 2023.Correction to: In the version of the article initially published, the affiliation of Ryan Cheng was incorrect and has now been updated to the Department of Pathology and Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, NY, USA, in the HTML and PDF versions of the article."} +{"text": "Schizophrenia is a chronic, frequent, and disabling psychiatric condition. The prognosis is more severe in the absence of treatment.The aims of our study were to evaluate the quality of treatment adherence and the quality of insight of patients with schizophrenia and to assess the implication of these factors as predictors of poor adherence.We conducted a cross-sectional and analytical study. We recruited 150 patients with schizophrenia treated at Razi Hospital of Manouba, divided into 113 patients with good adherence compared to 37 patients with poor adherence. We used the Medical Adherence Report Scale (MARS) to assess the quality of therapeutic adherence and the Birchwood Insight Scale for Insight Assessment.Poor treatment adherence in patients with schizophrenia was significantly associated with poor insight (p=0.001). Good adherence was associated with positive perception of treatment effectiveness (p<0.001). The predictive factor for poor adherence to therapy in multivariate analysis, after adjusting for the confounding variables was the negative perception of side effects (p=0.02). The predictive factor for good adherence was the presence of insight into the need for treatment (p=0.002).To prevent poor treatment adherence, a systematic screening for predictive factors and adequate management of schizophrenia would be imperative.A. Rami Shareolder of: no, Grant / Research support from: no, Consultant of: no, Employee of: no, Paid Instructor of: no, Speakers bureau of: no, E. Sana Shareolder of: no, Grant / Research support from: no, Consultant of: no, Employee of: no, Paid Instructor of: no, Speakers bureau of: no, C. Mejda Shareolder of: no, Grant / Research support from: no, Consultant of: no, Employee of: no, Paid Instructor of: no, Speakers bureau of: no, D. Rahma Shareolder of: no, Grant / Research support from: no, Consultant of: no, Employee of: no, Paid Instructor of: no, Speakers bureau of: no"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.1As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.3Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.5Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.7Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.16Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.22Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.10In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "In the present scenario, Alzheimer\u2019s Disease (AD) is one of the incurable neuro-degenerative disorders, which accounts for nearly 60% to 70% of dementia cases. Currently, several machine-learning approaches and neuroimaging modalities are utilized for diagnosing AD. Among the available neuroimaging modalities, functional Magnetic Resonance Imaging (fMRI) is extensively utilized for studying brain activities related to AD. However, analyzing complex brain structures in fMRI is a time-consuming and complex task; so, a novel automated model was proposed in this manuscript for early diagnosis of AD using fMRI images. Initially, the fMRI images are acquired from an online dataset: Alzheimer\u2019s Disease Neuroimaging Initiative (ADNI). Further, the quality of the acquired fMRI images was improved by implementing a normalization technique. Then, the Segmentation by Aggregating Superpixels (SAS) method was implemented for segmenting the brain regions , Mild Cognitive Impairment (MCI), Early Mild Cognitive Impairment (EMCI), Late Mild Cognitive Impairment (LMCI), and Significant Memory Concern (SMC)) from the denoised fMRI images. From the segmented brain regions, feature vectors were extracted by employing Gabor and Gray Level Co-Occurrence Matrix (GLCM) techniques. The obtained feature vectors were dimensionally reduced by implementing Honey Badger Optimization Algorithm (HBOA) and fed to the Multi-Layer Perceptron (MLP) model for classifying the fMRI images as AD, NC, MCI, EMCI, LMCI, and SMC. The extensive investigation indicated that the presented model attained 99.44% of classification accuracy, 88.90% of Dice Similarity Coefficient (DSC), 90.82% of Jaccard Coefficient (JC), and 88.43% of Hausdorff Distance (HD). The attained results are better compared with the conventional segmentation and classification models. AD is one of the leading chronic diseases, which generally affects people over 65 years . AD is aImplemented a normalization technique for improving the quality of raw resting state fMRI image by adjusting its contrast. The resultant image superiorly differentiates both bright and dark regions;Developed SAS methods for tissue segmentation such as AD, NC, MCI, EMCI, LMCI, and SMC. The SAS method partitions the denoised resting state fMRI image into multiple segments . The primary objective of the SAS method is to alter the image representation into perceptual meaning;Performed hybrid feature extraction (combination of Gabor and GLCM features) in order to extract vectors. Introduced HBOA for optimizing the dimensions of the extracted vectors. The feature extraction and optimization significantly reduce the number of redundant vectors that decreases the model\u2019s effort and increases the generalization steps and learning speed;Used MLP classifier to classify the tissues like AD, NC, MCI, EMCI, LMCI, and SMC. The MLP classifier has two main benefits in medical image classification; (i) effectively manage enormous amounts of data and (ii) resolves complex non-linear concerns. As depicted in the resulting segment, the efficacy of the presented model is zanalyzed in light of precision, HD, f1-measure, JC, accuracy, DSC, and recall.Among the available medical imaging methods, resting state fMRI is increasingly used for AD detection, because it is a useful tool for studying the brain, in terms of low-frequency fluctuation of Blood-Oxygen-Level-Dependent (BOLD) signal and alteration related to AD . The resIn this manuscript, the articles on the topic \u201cAD detection using raw resting state fMRI image\u201d are surveyed in Guo and Zhang presenteLi et al. have intRamzan et al. integratDuc et al. implemenSethuraman et al. implemenHojjati et al. initiallSun et al. developeJanghel and Rathore presenteZhang et al. developeOdusami et al. have devAnter et al. have intThe proposed system includes six phases for predicting AD in resting state fMRI images such as dataset description: ADNI, denoising: normalization technique, segmentation: SAS method, feature extraction: GLCM and Gabor features, feature optimization: HBOA, and classification: MLP model. The flow diagram of the proposed system is shown in In this manuscript, the proposed HBOA-MLP system\u2019s performance is tested on a benchmark dataset named ADNI . The ADNAfter acquiring the resting state fMRI images, the image denoising is accomplished by using the normalization technique. In this scenario, the normalization technique improves the quality of resting-state fMRI images by altering the contrast of the images ,33. The Input: Output: From the image The bipartite graph is constructed;The pixels are treated as the segment taken from the same group.The segmentation process is performed once the denoised resting state fMRI image is obtained. In this scenario, the aligned super-pixels are considered for image segmentation. Diverse patterns and multi-scale visual patterns are generally used for natural images. The super-pixels have a dissimilar combination of the cues to show promising results, but not fully explored. The super-pixels are collected from the SAS for partitioning, here; bipartite graph partitioning is linear to the pixels. The resultant images are negligible and constant compared to the super-pixels ,35. The After the calculation of SAS, the mean orientation is determined for the extracted resting state fMRI image. The information of the objects and the regions are merged, which provides detailed information for the mean shift and resultant segmentation. The process of mean shift performs clustering, and then the data point is calculated for every data that describes the mean shift. After describing the mean shift, image segmentation, tracking, mode seeking, visual tracking, etc. are performed. The obtained data points are fed to the most important and popular estimation methodology called kernel density, where the data points are represented as Hence, The kernel density is estimated to be superior using the mean square error that determines the optimal density. The density estimated in Equation (6) is rewritten and represented in Equation (7).The original density After segmenting the brain regions: AD, NC, MCI, EMCI, LMCI, and SMC from the denoised images, the feature extraction was performed by implementing GLCM and Gabor features. Initially, the Gabor features were calculated from the segmented fMRI images edundant . Therefo results . The undThe extracted 4463 vectors were fed to the HBOA for feature optimization. The HBOA follows the honey badger\u2019s behavior to catch the prey, and this optimization algorithm has two main steps (digging and honey) for resolving the optimization problems. In the honey step, the honey badgers follow honey birds for determining the beehive. In the digging step, the prey is determined based on the smelling ability of honey badgers ,39. FirsAfter the selection of 3260 vectors, MLP was employed for classifying the AD and its types such as AD, NC, MCI, EMCI, LMCI, and SMC. The MLP is one of the effective feed-forward neural networks, which includes benefits such as easy implementation and requiring only a small training set ,41. GeneThe presented segmentation method (SAS) and classification model (HBOA-MLP) were executed on a Matlab 2022a environment with a system configuration of 128 GB RAM, Windows operating system, 4 TB hard-disk, and Intel Core i9 14th-generation processor. Here, the presented SAS method and HBOA-MLP model\u2019s performance were tested on an online dataset: ADNI by means of JC, HD, DSC, f1-measure, recall, precision, and accuracy. Particularly, the SAS method\u2019s efficacy was analyzed in light of JC, HD, and DSC.The DSC was utilized for comparing the pixel-wise agreements between the ground-truth images and the segmented images. On the other hand, the JC is one of the effective functions that superiorly estimates the similarity measure between two resting-state fMRI images. In addition to this, HD is a useful and informative evaluation measure used extensively in medical image segmentation. The mathematical formulas of JC, HD, and DSC are mentioned in Equations (20)\u2013(22).Correspondingly, the classification performance of the HBOA-MLP model is analyzed in light of f1-measure, recall, precision, and accuracy. The f1-measure is a combined evaluation measure, which effectively captures the trade-off associated with recall and precision values. The evaluation measure: precision is defined as the proportion of classified positive cases, which are actually the real positive values.On the other hand, recall is defined as the proportion of actual positive classes, which are precisely classified. Lastly, accuracy is defined as the ratio of the total number of predictions to the number of correct predictions. The formulas used to compute accuracy, f1-measure, recall, and precision are given in Equations (23)\u2013(26).In the initial phase, the quantitative results of the proposed SAS method and other comparative segmentation methods are stated in The experimental results of the HBOA-MLP model and other comparative classification models are mentioned in On the other hand, in the case of HBOA, the MLP classifier obtained an f1-measure of 99.55%, precision of 99.28%, recall of 99.55%, and accuracy of 99.44%, which are superior to comparative classification models. Here, the experimental investigation is performed with 80:20% of training and testing of resting-state fMRI images. The visual presentation of the HBOA-MLP model and other comparative classification model results are specified in The experimental results of HBOA and other optimization algorithms are stated in The efficacy of the HBOA-MLP model was validated with the existing model: DRNN developed by Ramzan et al. , and it As mentioned in the introduction section, segmentation and feature optimization are integral parts of this research. The SAS significantly segments the brain regions from the denoised resting-state fMRI images. Further, the selection of optimal vectors decreases the system complexity to linear and the computational time of image classification to 62.11 s. However, the computational time of the HBOA-MLP model is superior to other comparative classification models and optimization algorithms. The efficacy of the presented segmentation method (SAS) and classification model (HBOA-MLP) are depicted in In this manuscript, a novel segmentation method (SAS) and classification model (HBOA-MLP) were proposed for the early diagnosis of AD using fMRI images. First, the quality of the collected fMRI images was enhanced by implementing the normalization technique, and, further, the brain regions were superiorly segmented by employing the SAS method. Next, the hybrid feature extraction and optimization were accomplished by utilizing Gabor features, GLCM features, and HBOA. The dimensionally reduced vectors were fed to the MLP classifier for image classification. In this scenario, the evaluation measures precision, HD, f1-measure, JC, accuracy, DSC, and recall were utilized for analyzing the efficacy of the segmentation method (SAS) and classification model (HBOA-MLP). As mentioned in the resulting segment, the HBOA-MLP model attained 99.44% of accuracy, and it is superior to the conventional comparative machine-learning models. On the other hand, the selection of optimal active vectors by HBOA diminished the proposed system complexity to linear and decreased the computational time of segmentation and classification to 42.33 s and 62.11 s. However, the MLP network includes too many parameters, because of its fully connected nature, and here, every node is connected with another node in a dense web that results in higher redundancy and inefficiency in the larger datasets. As a future extension, an effective transfer learning based CNN model is proposed with HBOA for precise AD prediction, because the MLP network is not ideal in processing patterns with multidimensional data. In addition, multimodal data , fMRI, and MRI) can be utilized for further enhancing the performance of AD prediction."} +{"text": "Nature 10.1038/s41586-023-06671-8 Published online 25 October 2023Correction to: In the version of the article initially published, the present address for Johannes Felsenberg was incorrect, and has now been updated to Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland in the HTML and PDF versions of the article."} +{"text": "Nature Communications 10.1038/s41467-023-40782-0, published online 19 August 2023Correction to: The original version of this Article contained an error in Fig. 3, panels A and B, in which the legend 'Mean- Proportion of Identical Prediction' was erroneously associated to the blue symbols and shading, instead of orange, and the legend 'Mean\u2014Tanimoto Similarity' was associated to the orange symbols and shading, instead of blue. This has been corrected in both the PDF and HTML versions of the Article."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Scientific Data 10.1038/s41597-022-01695-7, published online 12 October 2022.Correction to: The list of members of the Fibr Community Science Consortium in the original version of the paper incorrectly omitted consortium member Matthew D. Sacchet, Massachusetts General Hospital, Harvard Medical School, Boston, USA. The corrected membership list has replaced the incorrect version in the pdf and HTML versions of the article."} +{"text": "Nature 10.1038/s41586-023-05726-0 Published online 1 March 2023Correction to: In the version of this article initially published, the affiliation listed for David Caramelli was incorrect . The affiliation has been corrected to the Department of Biology, University of Florence, Florence, Italy in the HTML and PDF versions of the article."} +{"text": "Although a growing number of studies have attempted to uncover the relationship between plasma lipids and the risk of aortic aneurysm (AA), it remains controversial. Meanwhile, the relationship between plasma lipids and the risk of aortic dissection (AD) has not been reported on. We conducted a two-sample Mendelian randomization (MR) analysis to evaluate the potential relationship between genetically predicted plasma levels of lipids and the risk of AA and AD. Summary data on the relationship between genetic variants and plasma lipids were obtained from the UK Biobank and Global Lipids Genetics Consortium studies, and data on the association between genetic variants and AA or AD were taken from the FinnGen consortium study. Inverse-variance weighted (IVW) and four other MR analysis methods were used to evaluate effect estimates. Results showed that genetically predicted plasma levels of low-density lipoprotein cholesterol, total cholesterol, or triglycerides were positively correlated with the risk of AA, and plasma levels of high-density lipoprotein cholesterol were negatively correlated with the risk of AA. However, no causal relationship was found between elevated lipid levels and the risk of AD. Our study revealed a causal relationship between plasma lipids and the risk of AA, while plasma lipids had no effect on the risk of AD. Aortic aneurysm (AA) is the second most common aortic disease after atherosclerosis with a high risk of sudden death characterized by localized progressive and irreversible full-thickness dilation of the aorta . The incDyslipidemia refers to an imbalance in plasma levels of cholesterol and triglyceride, including total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglyceride (TG), and manifests as elevated plasma concentrations of TC, LDL-C, or TG, or a low plasma concentration of HDL-C or a combination of these features . NumerouIn recent years, due to the susceptibility of traditional observational studies to residual confounding effects and reverse causality, Mendelian randomization (MR) analysis has been widely used to evaluate the potential causality between various exposures and clinical outcomes by using genetic variants as instrumental variables (IVs) for exposures. Because the genetic variation occurs during conception and precedes the onset of disease, MR analysis can overcome reverse causation bias, reduce potential unmeasured confounders, and improve causal inference . The ranIn this study, we aimed to apply a two-sample MR analysis to investigate the role of genetically predicted plasma LDL-C, TC, TG, and HDL-C levels in the risk of AA and AD.We conducted a two-sample Mendelian randomization (MR) analysis to clarify the potential causal effects of genetically predicted plasma LDL-C, TC, TG, and HDL-C levels on AA and AD . Three kn = 440,546), TG , and HDL-C were obtained from the UK Biobank study, and plasma levels of TC were obtained from the GLGC study, which assessed the relationship between 4 lipid traits and the SNPs. SNPs associated with AA or AD were analyzed from FinnGen consortium data. Because the GWASs data are all publicly available and have been approved by the appropriate ethical review boards, no additional ethical approval was required for the analysis in this study.The analysis was carried out using published summary statistics from publicly available genome-wide association studies (GWASs), mainly on European individual characteristics, both male and female . The UK p < 5 \u00d7 10\u22128) were selected. Second, the independence of these selected SNPs was evaluated based on the linkage disequilibrium. SNPs were deleted if they were in the linkage disequilibrium (r2 > 0.01) and in proximity (clumping window of 1 Mb) to other SNPs with higher p-values. Third, SNPs with F-statistics greater than ten were selected to mitigate the impact of potential bias.Independent SNPs associated with plasma levels of LDL-C, TC, TG, or HDL-C were identified according to three criteria. First, SNPs that had reached a genome-wide significance level , MR-Egger, weighted median, simple mode, and weighted mode, were applied in our study. Among the five methods, the IVW method was used as the primary analysis method to evaluate the effect estimates. Cochran\u2019s Q test was applied to test the heterogeneity of the IVs, and significant heterogeneity was considered to exist when p value < 0.05 was considered as statistically significant. All statistical analyses were performed using the \u201cTwoSampleMR\u201d packages in R Software (version 4.2.0).Two-sided Independent SNPs included in our study as IVs are shown in p = 0.006), TC , or TG were associated with an increased risk of AA. Meanwhile, an elevated plasma level of HDL-C was associated with a decreased risk of AA , 1.47; 95% confidence interval (CI), 1.20\u20131.81; sk of AA . These ask of AA . The scask of AA , so the sk of AA . The leask of AA . Considesk of AA .p = 0.542), LDL-C , TC , or TG was not associated with the risk of AD (The IVW analysis revealed that the genetically predicted plasma level of HDL-C (OR, 0.90; 95% CI, 0.64\u20131.26; sk of AD . Given tsk of AD .In this two-sample MR analysis, we investigated the genetic association between levels of plasma lipids and the risk of AA and AD. Our study provided new evidence for the previous clinical findings that elevated plasma levels of LDL-C, TC, or TG were associated with an increased risk of AA, and an elevated plasma level of HDL-C was inversely associated with the risk of AA. In addition, we revealed for the first time that, based on the same GWASs databases, there was no causal relationship between genetically predicted lipid levels and the risk of AD.Current studies on the relationship between plasma lipids and the risk of AA are still controversial. An observational study revealed that LDL-C, TC, TG, and HDL-C were independently associated with the risk of AAA . HoweverAt present, there are no studies on the association between plasma lipids and the risk of AD. Using MR analysis, we found, for the first time, that genetically determined plasma levels of HDL-C, LDL-C, TC, and TG were not associated with the risk of AD. Using the same IVs from the UK Biobank and GLGC studies and the same population from the FinnGen consortium database, there were positive results for AA but negative results for AD, which greatly enhanced the reliability of our findings on estimates of causality between genetically determined plasma lipid levels and the risk of AD and indicated that the pathogenesis of AA and AD were very different. An observational study found that patients with AD combined with basic atherosclerotic diseases had significantly higher levels of LDL-C, TC, and TG and significantly lower levels of HDL-C in their plasma than patients with basic atherosclerotic diseases alone . There aA previous two-sample Mendelian randomization study, based on the GLGC and UK Biobank databases, assessed whether there was a gender-dependent difference in the effect of genetically determined plasma levels of LDL-C on the risk of AA and AD . In the Lipid-lowering therapy is considered to play an important role in the prevention of a variety of cardiovascular diseases, such as coronary heart disease, myocardial infarction, and stroke ,29,30,31In our study, we used a two-sample MR analysis based on the large-scale GWASs databases to evaluate the causal effect of genetically predicted plasma lipid levels on the risk of AA and AD. Compared with traditional observational studies, our study overcame the influence of residual confounding effects and reverse causality, which enhanced the reliability of our findings. Using different databases from those used in previous studies, our study provided new evidence for the nature of the causal relationship between genetically determined lipid levels and the risk of AA. Meanwhile, we revealed for the first time that genetically determined lipid levels are not associated with AD risk. However, several limitations of our study should be considered. First, all large-scale GWASs data used in our study were obtained from the European population. Therefore, whether our findings are applicable to other ethnic groups requires further study. Second, the data on association between the genetic variant and AD or AA were taken from the FinnGen consortium database. Aortic aneurysms and dissections were not classified into different types in this database, so the effect of dyslipidemia on different types of aneurysms and dissections could not be obtained. Finally, although no evidence of pleiotropic effects was found in the MR-Egger intercept tests from our study, the potential influence of directional pleiotropy is difficult to completely exclude.In conclusion, our study revealed a causal relationship between genetically determined plasma lipid levels and the risk of AA, and found for the first time that genetically determined plasma lipid levels had no effect on the risk of AD, which may help to guide clinical prevention, clinical trial design, and risk-factors analysis of AA and AD."} +{"text": "Four categories of e-Health modalities were identified: web-based (n = 19), mobile application (n = 3), virtual reality (VR) (n = 2), and video consulting (n = 2). Interventions were mainly based on the cognitive behavioral therapy (CBT) approach (n = 14) and mostly involved contact with a healthcare professional through different digital tools. Overall, a growing number of psychological and multicomponent interventions have been created and delivered using digital tools in the context of FMS, showing their potentiality for improving psychosocial outcomes and pain-related psychological variables. However, some digital tools resulted as underrepresented, and the literature on this topic appears highly heterogeneous precluding robust conclusions.There is growing evidence to support the potential benefit of e-Health interventions targeting psychosocial outcomes and/or pain-related psychological variables for chronic pain conditions, including fibromyalgia syndrome (FMS). This systematic review aims at providing an in-depth description of the available e-Health psychological and/or multicomponent interventions for patients with FMS. Searches were made in PubMed, Cochrane, Web of Science, and PsycINFO up to 15 May 2023, finally including twenty-six articles. The quality of the included articles was medium\u2013high . 50% of studies were randomized controlled trials (RCTs) ( Fibromyalgia syndrome (FMS) is a chronic disease characterized by widespread musculoskeletal pain and associated with other highly disabling symptoms such as fatigue, poor sleep, cognitive dysfunction, depression, and anxiety , resultiThe range of e-Health interventions and digital applications is broad and continues to evolve. Different e-Health tools have been introduced and created for chronic pain management. Slattery and collaborators includedTo the best of our knowledge, until now, only one systematic review has focused specifically on IPTs for FMS patients . The autTo provide a systematic and in-depth description of the available e-Health tools delivering multicomponent and psychosocial interventions targeting psychosocial outcomes and/or pain-related psychological variables for patients with FMS;To describe the main psychological approaches used in those e-Health interventions, their structure, and their main characteristics, along with the main psychosocial outcomes and pain-related psychological variables targeted in the included interventions;To describe the impact of the e-Health tools in terms of signals of efficacy, feasibility, and acceptability.Considering that the body of the research literature in the field of e-Health tools has resulted in higher and broader interest in the past five years, and in particular after the recent pandemic, the current paper updates and enlarges the scope of this previous systematic review, including all the potential e-Health psychological and multicomponent interventions targeting psychosocial outcomes and pain-related psychological variables for patients with FMS. Based on these premises, the aims of the current review are threefold:This review was carried out following the \u201cPreferred Reporting Items for Systematic reviews and Meta-Analyses\u201d (PRISMA) guidelines .What e-Heath tools are under investigation to deliver psychological and/or multicomponent interventions targeted psychosocial outcomes and/or pain-related psychological variables in patients with FMS?What are the main characteristics of those e-Health interventions in terms of underlying psychological approaches, structure, and addressed outcomes?What is the impact of such e-Health tools in terms of signals of efficacy, feasibility, and acceptability?Three research questions guided the current review:We systematically searched four electronic databases up to 15 May 2023. We used the following search strategy: AND , we included only the first published study. However, when relevant data were reported in subsequent publications , these papers were excluded, but, if present, the relevant information was considered in the narrative synthesis of the results.The following types of studies were excluded: systematic review, narrative review, meta-analysis, bibliometric analysis, letter, case-study, book/book chapter, comment, editorial, congress abstract or symposium, poster presentation, and dissertation.All eligible studies were evaluated against the 16-item quality assessment tool (QATSDD) . The tooA systematic and in-depth description of the following data was conducted: study design; characteristics of the sampled population for age, gender, and presence of psychiatric diagnosis among the criteria; the intervention outcomes and pain-related psychological variables; the follow-up duration of the study, when applicable; when involved, the type of control group extracted data from the selected studies using a data-collection form in Microsoft Excel. Doubts were discussed, and any disagreement about study eligibility was resolved by a third reviewer (V.D. or C.P.).The electronic literature search yielded 2698 records in total, with 777 duplicates that were removed. During the study selection process, 1921 records were analyzed by title and abstract, and 1831 were excluded according to the inclusion and exclusion criteria. Finally, 92 records were selected for the full-text analysis, of which 66 were excluded for various reasons , while the subsequent papers were excluded from the flowchart and whenTwenty-six studies finally met the inclusion criteria ,51,52,53n = 13) were randomized controlled trials (RCTs) )To conclude, some issues remain open and thus need to be addressed. In particular, further studies should fill the knowledge gap about the clinical characteristics and the most effective treatment options for FMS when it occurs in specific populations, including males, young and old individuals, and patients with psychiatric comorbidities. As for the latter aspect, there is a lack of consistency between the included studies regarding the inclusion/exclusion of patients with psychiatric diseases, generally without providing any reason for this choice. This issue should therefore be addressed in future investigations. Moreover, e-Health potential solutions other than web-based modalities seem underdeveloped in this field, suggesting the need for future research into them. Finally, feasibility parameters and cost evaluations of such tools should be taken into account by future studies.This systematic review presents both strengths and limitations. Regarding its strengths, this systematic review enlarges and updates the results of a previous review on the topic, allowing for a deeper analysis and understanding of the potentiality of e-Health tools for psychosocial benefits for patients with FMS. The rigorous methods used for the search strategy, data analysis, and studies appraisal were based on internationally recognized tools.Regarding its limitations, the high heterogeneity of the included studies in most of the targeted variables of the review deeply limited our possibility to draw conclusive results. Moreover, the classification and description of the interventions were based on the information reported in the studies, which was often limited. As suggested by Rohn et al. , a more The present review presents a complete description of the state of the art on use of e-Health strategies targeting psychosocial outcomes and pain-related psychological variables for patients with FMS.A growing number of psychological and multicomponent interventions using e-Health tools in the context of FSM have emerged, with a significant prevalence of interventions based on a web-based modality. Still, few experiences of m-Health, VR, and video consulting have been implemented in the FMS context. Looking at the extensive range of psychosocial variables targeted by the e-Health tools and the signals of the efficacy of the included interventions, e-Health tools have shown the potential to positively influence psychosocial variables representing the core dimensions of FMS.We also showed that the existing literature is highly heterogeneous regarding study design, psychosocial outcomes, instruments to assess psychosocial dimensions, and e-Health tools reported across studies, making it difficult to provide robust conclusions. Far from being merely a limitation, such an observation can stimulate a discussion on the need to develop further research in FMS and e-Health intervention. Therefore, future studies should accurately focus on selecting variables and instruments to improve the comparison among results."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.1As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.3Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.5Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.7Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.16Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.22Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.10In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Dear Editor,The Middle East respiratory syndrome coronavirus, often known as MERS-CoV, is a new emerging beta coronavirus that belongs to lineage C. About 80% of all human cases of this virus have been reported to Saudi Arabia, and more than 2600 confirmed cases have been diagnosed in humans with at least 1000 fatalities globally since it was first detected in 2012According to a recent disease outbreak statement issued by the WHO, there have been four laboratory-confirmed cases of MERS-CoV documented from three different areas inside the Kingdom of Saudi Arabia. WHO revealed in April 2022 that the Kingdom of Saudi Arabia had been the source of six prior MERS-CoV infections, including four fatalities, between August 2021 and February 2022. The most current epidemic includes three cases that occurred between 29 December 2021, and the end of October 2022. Two cases originated in Riyadh, while the other came from Gassim and Makka Al Mukarramah. Real-time PCR confirmed the instancesThe MERS-CoV genome is estimated to have 10 open reading frames (ORFs) and that the first 5\u2032-three-fourths of the sequence encode for RdRp (ORF1a and ORF1b)Besides, research has found that the molecular epidemiology of MERS-CoV is similar to the initial human cases, which were first diagnosed in 2012. The current molecular features of the MERS-CoV virus are ~99% similar to the sequences observed in comparison to the first human cases, and the level of pathogenicity is also reported as the sameThe MERS-CoV virus is a zoonotic pathogen, meaning it may be spread from animals to humans. Although the precise transmission mode is unknown, research has found that people get sick via contact with diseased dromedary camels. Several Middle Eastern, African, and South Asian Member States have confirmed the presence of MERS-CoV in dromedary camelsTransmission between humans is feasible and has already happened, most often between intimate friends and family members, particularly in medical settings. Many others might be affected, such as members of the patient\u2019s immediate family, home, and healthcare providers. Most of the disease has spread via hospitals in Saudi Arabia, the United Arab Emirates, and the Republic of Korea. There is no evidence of persistent human-to-human transmission outside healthcare settingsDiagnostics are crucial in the early identification and management of any pathogenic disease. They also offer a more comprehensive knowledge of the epidemiological data and risk factors for MERS-CoVThe WHO has provided guidelines for treating severe respiratory illnesses likely caused by the MERS-CoV virus. Nevertheless, there is currently no antiviral therapy that is specifically indicated for the treatment of MERS-CoV infectionNot applicable/not required. This is correspondent does not require any human or animal subjects to acquire such approval.This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.S.A., N.M.: conceptualization, study design, and writing. S.A., F.I.A., S.M., and N.M.: writing and analyzed the data. F.I.A., S.M., N.M., and A.A.: editing and reviewing.Authors declare no conflict of interest exists.Name of the registry: not required.Unique identifying number or registration ID: not required.Hyperlink to your specific registration (must be publicly accessible and will be checked): not required.Shopnil Akash (corresponding author) takes full responsibility for the work and/or the conduct of the study, has access to the data, and controls the decision to publish.Extensive research into MERS-CoV is needed to ascertain its current status, epidemiological context, and potential in the future."} +{"text": "Recent research evidence has shown the importance of different psychological constructions for analyzing problems associated with lack of adequate behavior management in human beings. The model of the different levels of behavioral analysis \u2013 microanalysis level, molecular level and molar level \u2013 allows us to approach the study of executive functions, in relation to other constructions of said levels have become an essential construct for explaining learning difficulties and self-regulation of behavior in the lives of individuals. This well-documented construct represents the level of microanalysis of human behavior, which means that it focuses on the interaction between brain and behavior on cognitive performance, including decision-making across the lifespan. In parallel and complementary, other psychological models from research at the molecular and molar levels have been developed aiming to fill out the analysis and definition of behavior regulation, especially in the educational and health fields. A paper analized \u201cThe executive function and effortful control, with similar and different evidence from big data analysis\u201d (Chae). Another article has analyzed the \u201cIce Cream,\u201d a new virtual reality tool for the assessment of executive functions in children and adolescents, with a normative study .1) On the one hand, the Self-Regulated Learning, SRL, and general Self-Regulation modeled after the information processing paradigm have enabled us to accurately understand self-regulatory processes in the human being. These models, placed at a molecular level of analysis, give us a sequential, discrete understanding of self-regulatory behaviors, in the sphere of education and health. By analyzing across the behavioral sequence of before-during-after each act, the models have provided evidence of their value in assessment and intervention. Two works analyze the effect of Executive Functioning on school learning, both in Reading Comprehension (Leshem and Altman), and in English (Akhmedjanova and Moeyaert). Additionally, three articles have focused on the effect of self-regulated learning: the first, in asynchronous online learning situations ; the second report has analyzed the combined value of executive functions and self-regulated learning to predict differences in study success within higher education students ; a third work focused on noise reduction in preschool from a self- regulated learning perspective\u2014implementation of a game-based voice regulation training program (Sarfaty and Ben-Eliyahu).2) The behavioral models of Self- vs. External Regulation, SR vs. ER Theory takes its place at the molar level of analysis and has postulated the relevance of an interactive subject x environment analysis. This model has confirmed the relevance and value of the interaction of levels of regulation present in the subject and in their context, for predicting human behaviors in the fields of education and health. The continuum of Self-Regulation, Non-Regulation, Dys-Regulation (SR-NR-DR) has helped to operationally define the types of regulatory behavior, whether at the personal level or the contextual level. Two works focused on the effect of context on regulation have analyzed the factors affecting faculty conformity in South China universities (Xu and Chang) and the importance of teachers' supportive vs. undermining behavior for developments in early adolescents' self-regulation (Opdenakker). Additionally, two research reports have provided evidence regarding the combination of predictive factors of contextual and personal regulation regarding the variability of executive functions in university students and the learning specific regulatory behavior .3) Finally, the behavioral model of In conclusion, this Research Topic has provided a multilevel view of the executive functions construct, in relation to other related constructs, from a multilevel perspective. Future work should delve into this problem, since it is neither closed nor exhausted.JF: Conceptualization, Funding acquisition, Investigation, Project administration, Writing \u2013 original draft. LF: Conceptualization, Supervision, Writing \u2013 review & editing. FS: Resources, Supervision, Validation, Writing \u2013 review & editing. MP: Formal analysis, Methodology, Writing \u2013 original draft. UD-O: Conceptualization, Methodology, Validation, Writing \u2013 review & editing."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "In \u201cEfficacy, Effectiveness, and Quality of Resilience-Building Mobile Health Apps for Military, Veteran, and Public Safety Personnel Populations: Scoping Literature Review and App Evaluation :e26453]) the authors made the following 4 corrections:1. In the originally published article, in 8 instances the name of an app appeared as:Resilience@Work/MindarmaThis has been corrected as follows in all 8 instances:Mindarma2. In the originally published article, in the Results: Study Findings: Evidence-Based Merit section, the last sentence appeared as follows:Virtual Hope Box, eQuoo, and Resilience@Work/Mindarma were evaluated separately in their respective RCT studies.This has been corrected as follows:Virtual Hope Box and eQuoo, were evaluated separately in their respective RCT studies. It was noted that Mindarma was utilized as a part of a mindfulness program for first responders .3. In the originally published article, in the Results: Study Findings: Mental Control, Emotional Regulation, Coping, and Self-efficacy section, the following sentence appeared:Resilience@Work/Mindarma was the only app in this study that drew from acceptance and commitment therapy principles.This sentence has been deleted from the paper.4. In the originally published article, in the Results: Study Findings: Effect of Apps on Resilience section, the following sentences appeared:Similarly, Resilience@Work/Mindarma showed improved adaptive resilience and psychological flexibility . Joyce eThese sentences have been deleted from the paper.The correction will appear in the online version of the paper on the JMIR Publications website on August 28 2023, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "Significant heat-related casualties underlie the urgency of establishing a heat-health warning system (HHWS). This paper presents an evidence-based pilot HHWS developed for Taipei City, Taiwan, through a co-design process engaging stakeholders. In the co-design process, policy concerns related to biometeorology, epidemiology and public health, and risk communication aspects were identified, with knowledge gaps being filled by subsequent findings. The biometeorological results revealed that Taipei residents were exposed to wet-bulb globe temperature (WBGT) levels of health concern for at least 100 days in 2016. The hot spots and periods identified using WBGT would be missed out if using temperature, underlining the importance of adopting an appropriate heat indicator. Significant increases in heat-related emergency were found in Taipei at WBGT exceeding 36\u00b0C with reference-adjusted risk ratio (RaRR) of 2.42, taking 30\u00b0C as the reference; and residents aged 0\u201314 had the highest risk enhancement (RaRR = 7.70). As for risk communication, occurring frequency was evaluated to avoid too frequent warnings, which would numb the public and exhaust resources. After integrating knowledge and reconciling the different preferences and perspectives, the pilot HHWS was co-implemented in 2018 by the science team and Taipei City officials; accompanying responsive measures were formulated for execution by ten city government departments/offices. The results of this pilot served as a useful reference for establishing a nationwide heat-alert app in 2021/2022. The lessons learnt during the interactive co-design processes provide valuable insights for establishing HHWSs worldwide. In 2022, extreme weather events under the trend of climate change have caused thousands of deaths in the US, Europe, and Asia . In ordeWith the aim of providing solutions, it is crucial to not only integrate evidence and knowledge during iterative discussions among the involved multidisciplinary scientists and stakeholders but also reconcile different preferences and perspectives in the co-design process. Nevertheless, apart from a book released in 2021 , very feThe significant worldwide heat-related casualties in recent years have highlighted the urgency and importance of establishing an effective HHWS , 10\u201317. The fact that any developed HHWS should be region-specific was acknowledged by the Guidance on Warning-System Development put forward by the World Meteorological Organization (WMO) and the World Health Organization (WHO) . In addiIn the city-level pilot work presented here, WBGT was chosen as the heat indicator for multiple reasons. As a weighted combination of dry-bulb temperature, natural wet-bulb temperature, and globe temperature, WBGT considers four heat-stress relevant meteorological parameters, including temperature, relative humidity, wind speed, and solar radiation . WBGT isThe objective of this paper is to showcase an evidence-based pilot HHWS developed for Taipei City, Taiwan, through a co-design process, which is presented to highlight the viewpoints and preferences of policy makers. The scientific findings are also summarized to address the policy concerns corresponding to the aspects of biometeorology, epidemiology and public health, and risk communication identified in the co-design process. The overall considerations in integrating scientific evidence with policy concerns from different perspectives are also presented. With certain detours, the knowledge acquired from this pilot work of 2018 has laid a foundation for the nationwide heat-alert app developed in 2021/2022, demonstrating the difficulty involved in transforming knowledge into actions for tackling the real-world challenge of sustainability. Lessons learnt in the process can be applied to other countries and facilitate the implementation of HHWSs for reducing health risks to meet the United Nation Sustainable Development Goal 3 (SDG3), ensure healthy lives and promote well-being for all at all ages . The essThe co-design process involving stakeholder engagement, the methodology for addressing three aspects of the policy concerns of this pilot HHWS, and the implementation of this pilot HHWS along with the planning of responsive actions are described in this section.Two key stakeholders were involved in this process, one of which is the Central Weather Bureau (CWB), with the mandate of developing the nationwide HHWS. In view of the higher mortality and morbidity on hot days in Taiwan , 35, 42,The other stakeholder is the Department of Environmental Protection of Taipei City Government (TDEP), which is responsible for reducing impacts of climate change. In 2017, upon learning about WBGT from the CWB, the TDEP officials initiated discussion with our team and were later convinced of the suitability of WBGT as the heat indicator for an HHWS, especially for people engaged in outdoor activities. A pilot HHWS was thus developed for Taipei City (abbreviated as \u201cTPE-pHHWS\u201d), with science-based criteria and a mechanism set up for triggering responsive programs within the Taipei City Government to reduce heat-health risks. Furthermore, the underlying anticipated benefit is to gain knowledge that may induce and facilitate the CWB to establish a national WBGT-based HHWS.The knowledge gaps were identified as the first step in developing this WBGT-based TPE-pHHWS. The major policy concerns regarding three aspects of warning-system development emphasized by WMO and WHO, namely, biometeorological, epidemiological and public health, and risk communication, are shown in TM, Oconomowoc, WI, USA), which is not waterproof and thus not suitable for long-term outdoor monitoring. Instead, our team obtained WBGT using equations derived by scientists in the US national laboratories according to fundamental principles of heat and mass transfer with inputs of routine meteorological measurements, which has been validated with monitoring under a CC BY license, with permission from Department of Land Administration, Ministry of the Interior, Taiwan, original copyright 2016. Township and urban boundaries were obtained from open data [https://data.gov.tw/dataset/7441] under a CC BY license, with permission from National Land Surveying and Mapping Center, Ministry of the Interior, Taiwan, original copyright 2015.The blue and black lines show the border of Taipei City and the entire Taipei metropolitan area, respectively; the magenta and purple dots are 53 and 4 stations in elementary schools at altitude < and \u2265 100 m, respectively; the star signs indicate the four HOBO monitoring stations of the TPE-pHHWS. Grid numerical terrain model data (20-meter resolution) were obtained from open data [(TIF)Click here for additional data file.S2 FigNumber of days above thresholds for (a) Japan WBGT categories for the general public and (b) US WBGT categories for the workers. Number of days in the highest WBGT category are listed on the top of the respective column.(TIF)Click here for additional data file.S3 Fighttps://www.cwb.gov.tw/V8/E/P/Warning/W29.html (English)) and (b) the classification of Wet-bulb Globe Temperature (WBGT) translated from the data description of https://opendata.cwb.gov.tw/dataset/forecast/M-A0085-001 in Mandarin posted by Central Weather Bureau. Note: The Central Weather Bureau has been upgraded to the Central Weather Administration on September 15, 2023; thus, the webpage may be renamed as www.cwa.gov.tw in the future.(a) Classification of temperature according to \u201cheat information\u201d announced by Central Weather Bureau ((TIF)Click here for additional data file."} +{"text": "There are errors in the Funding section. The correct Funding statement is: This research was supported by the National Evidence-based Collaborating Agency, Republic of Korea (grant number: NA22-008 and NA23-010). There was no additional internal or external funding received for this study. The funding source had no role in the study design, data collection, and analysis, decision to publish, or manuscript preparation of the manuscript."} +{"text": "Atopic dermatitis (AD), often called eczema, is characterized by intense pruritus, erythema, dry skin, and inflammation. The condition is chronic and relapsing, and often occurs in patients with a family history of the atopic triad . Use of topical steroids has been the mainstay of medical treatment for AD. Steroid-free treatments for AD, with a more favorable safety profile, have become available within the past 2 years. Tacrolimus ointment, a topical immunomodulator, became available in early 2001 and is indicated for moderate-to-severe AD. A similar but highly skins elective cytokine inhibitor, pimecrolimus cream 1%, became available in March 2002. Pimecrolimus is indicated for mild-to-moderate AD. The objective of this article is to review the key characteristics that differentiate pimecrolimus from steroids and tacrolimus in the treatment of AD.Using secondary resources, the clinical aspects and conventional treatment strategies for AD are reviewed as are the pivotal clinical studies with pimecrolimus and literature on quality of life and economic burden of disease for AD patients and families.Pimecrolimus is an effective, steroid-sparing therapy for mild-tomoderate AD. Early treatment prevents flares, the agent works quickly to reduce signs and symptoms of more advanced AD, and it is safe and appropriate for intermittent long-term therapy. Pimecrolimus has fewer side effects than topical steroids and a better side-effect profile than tacrolimus. It can also be used as a first-line therapy. In studies with patients aged 2 to 17 years, it has been shown to be particularly effective in improving eczema of the face and neck, and its use may improve quality of life for many patients, especially children. A single-strength dose (1%) is safe and medically beneficial for pediatric, adolescent, and adult patients. The direct drug cost of pimecrolimus compares favorably with tacrolimus, but it is significantly more expensive than generic topical steroid creams."} +{"text": "Inequity in maternal-child health services is a challenge to global health as it hinders the achievement of Sustainable Development Goals (SDGs) and Universal Health Coverage. Though the Association of Southeast Asian Nations (ASEAN) has made remarkable achievements in maternal-child health, there remain gaps in reaching global goals. This study aimed to compare and investigate the inequity in maternal-child health (MCH) services in ASEAN member states to help guide policy decisions to improve equitable health services in the SDG era and beyond.Using the WHO Health Inequality Monitor, we identified inequity summary measures for five MCH services in ASEAN member states from 1993 to 2021: antenatal care, births attended by skilled health personnel, diphtheria, tetanus and pertussis (DTP3) immunization, measles immunization, and polio immunization. We divided the analysis dimension of inequity into urban\u2013rural inequity, economic status inequity, and sub-regional inequity. Trends of absolute and relative inequity in every dimension of MCH services in ASEAN member states were examined with the principal component analysis (PCA).The mean coverages of MCH services are 98.80% (Thailand), 86.72% (Cambodia), 84.54% (Viet Nam), 78.52 (Indonesia), 76.94% (Timor-Leste), 72.40% (Lao PDR), 68.10% (Philippines) and 48.52% (Myanmar) in 2021. Thailand have the lowest MCH services absolute inequity indexes of -1.945, followed by Vietnam (-1.449). Lao PDR and Myanmar have relatively higher MCH services absolute inequity indexes of 0.852 and 0.054 respectively. The service in Cambodia, Indonesia, and the Philippines is pro-specific regions . The service in Myanmar is pro-rich (with economic status absolute inequity index of 0.43). The service in Lao PDR and Timor-Leste is pro-urban areas, pro-rich, and pro-specific regions.The inequity of MCH services in ASEAN persists but is in a declining trend. Thailand and Vietnam have performed well in ensuring MCH services equity, while Laos and Myanmar are still facing serious inequity dilemmas. The progress of MCH service equity in Myanmar, Cambodia, the Philippines, and Indonesia is uneven. It is acceptable to learn from the successful experiences of Thailand and Vietnam to improve the equities in other ASEAN countries. Policies should be developed according to the specific types of MCH inequity in member states to improve equity levels.The online version contains supplementary material available at 10.1186/s12939-023-01974-8. Maternal-child health (MCH) is at the core of the United Nations Sustainable Development Goals (SDGs) and universal health coverage (UHC) . Each stThe Association of Southeast Asian Nations (ASEAN), which consists of Negara Brunei Darussalam, the Kingdom of Cambodia, the Republic of Indonesia, the Lao People\u2019s Democratic Republic, Malaysia, the Republic of the Union of Myanmar, the Republic of the Philippines, the Republic of Singapore, the Kingdom of Thailand, the Socialist Republic of Vietnam and the Democratic Republic of Timor-Leste, has accomplished several notable achievements in economic and social development . As the th ASEAN Senior Officials Meeting for Health Development (SOMHD) endorsed the ASEAN Post-2015 Health Development Agenda (2021\u20132025) [Health Cluster 3: Strengthening Health System and Access to Care. Seen from the overall development, the equity of maternal-child health services in ASEAN member states is improving year by year. Some ASEAN member states have made remarkable achievements in MCH services in recent years. For instance, MMR in Laos has decreased from 357.0 in 2012 to 62.0 in 2021, and in the Philippines has decreased from 69.9 in 2013 to 56.8 in 2018. U5MR in Cambodia dropped from 45.2 in 2013 to 25.9 in 2020 [For the vision of healthy, caring, and sustainability, the 1621\u20132025) , in whic in 2020 .When there are marked inequities, countries which are disadvantaged may lack the resources to participate in the social and economic mainstream of global society . ImproviWe obtained data from the Thirteenth General Program of Work (GPW 13) indicators in the WSince the data set does not contain relevant data of Malaysia, Brunei, and Singapore, this study focused on member states including Cambodia, Indonesia, Laos, the Philippines, Thailand, Timor-Leste, Vietnam, and Myanmar. For this study, the inequity in maternal-child health services was measured by five indicators, including antenatal care coverage, births attended by skilled health personnel, diphtheria, tetanus and pertussis (DTP3) immunization coverage, measles immunization coverage, and polio immunization coverage. Years of data collection ranged from 1993 to 2021. Cambodia has five years of data for 2000, 2005, 2010, 2014, and 2021; Indonesia for 1997, 2002, 2007, 2012, and 2017; Laos for 2011 and 2017; Philippines for 1993, 1998, 2003, 2008, 2013, and 2017; Thailand for 2012, 2015, and 2019. Timor-Leste for 2009 and 2016; Vietnam for 1997, 2002, 2010, 2013, and 2021; Myanmar for 2015.difference(X) and ratio(Y) of service coverage between urban and rural areas, the richest and poorest wealth quintiles, and areas with the highest and lowest service coverage in each country each year. The Difference shows the absolute inequality between two subgroups and the ratio shows the relative inequality [For analyzing the degree of inequity by different dimensions robustly, we calculated summary measures including equality . The relThen we used the principal component analysis (PCA) to review the trends and types of the inequity in MCH services among ASEAN member states. PCA can classify multiple factors into fewer factors, fully reflect the original information, and the transformed factors have no linear correlation . PCA canAccording to the latest data from the WHO Global Health Observatory , the covFigure\u00a0The KMO values of the absolute difference between urban and rural areas, richest and poorest quintiles, and two subregions with the most extreme values in maternal-child health services are 0.776, 0.733, and 0.816 respectively . The significance of the Bartlett spherical test was less than 0.05. The data met the requirements of PCA. The eigenvalues and the proportion of explained variance are obtained after the principal component analysis of the third-level indicators under the second-level indicators. The principal components were selected according to the eigenvalue and proportion to calculate the general scores high overall-coverage high equity, (2) high overall-coverage structural inequity, (3) low overall-coverage structural inequity, and (4) low overall-coverage high inequity. Thailand and Viet Nam are characterized by type one. Cambodia and Indonesia are characterized by type two. The Philippines and Myanmar are characterized by type three. Lao PDR and Timor-Leste are characterized by type four.helping the poor is one of the important funding and subsidy projects [the Ministry of Health Plan for People\u2019s Health Protection, Care and Promotion 2016\u20132020 [In terms of both coverage and equity level of MCH services, Thailand is a model among ASEAN member states, which is inseparable from its well-established schemes include extensive geographical coverage of functioning primary health care and rural recruitment, home town placement, and financial and non-financial incentives to improve the availability of health workers in underserved areas , 34. In projects . On Marc016\u20132020 , in whic016\u20132020 . As one Although the overall service coverage of Cambodia and Indonesia is relatively high, they have inequity in health services in some dimensions. The service coverage in both Cambodia and Indonesia is pro-specific regions. Therefore, while maintaining the overall progress of maternal and child health service coverage, Cambodia and Indonesia should make targeted investments in balancing regional development.The overall service coverage of the Philippines and Myanmar is relatively low, and they also have inequity in health services in some dimensions. The service coverage in the Philippines is pro-specific regions, and Myanmar is pro-urban areas and pro-specific regions. It is worth noting that the inequity of regional MCH services in the Philippines has shown a worsening trend in recent years. The Philippine government may has recognized this issue , and it The overall level of health service coverage in Lao PDR and Timor-Leste is relatively low, and they have inequity in health services in all dimensions. The current health development plan in Laos was made to solve the cultural, financial, and geographical barriers faced by vulnerable groups in accessing health services, so as to achieve the full coverage of high-quality health services . During It is essential to Enhance the capacity of ASEAN member states, and the focus should be tilted to key countries. In ASEAN Region, five countries \u2013 Cambodia, Indonesia, Lao PDR, Myanmar, and the Philippines, remain high in maternal mortality . Timely At the level of ASEAN member states, it is essential to ensure policy decisions do not worsen the status of inequities , and to This study has several limitations. First, WHO Health Inequality Monitor does not contain all ASEAN member states, so comparative analysis can only be conducted among the eight member states with relevant data. Second, to include as many years, countries, and inequity dimensions as possible, we ultimately only selected five MCH indicators for analysis. It is necessary to explore and use more detailed data to conduct more in-depth research on the situation and causes of inequity in maternal and child health services in ASEAN countries in the future, like integrating service quality information based on crude service coverage. Third, the statistical data for some member states have not been updated for several year, meaning that there might be a change in the inequity pattern of maternal-child health services in these countries in recent years. (e.g. The COVID-19 pandemic may have magnified inequities and threatens to exacerbate them) .Equity analysis is vitally important to identify who gets the worst quality of care, which helps guide policy decisions toward equitable distribution of health resources in the SDG era and beyond . Our finAdditional file 1: Appendix Table 1. List of maternal and child health service equity indicators. Appendix Table 2.\u00a0 The eigenvalue and contribution proportion of every principal component. Appendix Figure 1. Trend of tertiary indicators (relative difference) in each country."} +{"text": "Schizophrenia is among the fifteen most disabling diseases worldwide. Negative symptoms (NS) are highly prevalent in schizophrenia, negatively affect the functional outcome of the disorder, and their treatment is difficult and rarely specifically investigated. Serotonin-dopamine activity modulators (SDAMs), of which aripiprazole, cariprazine, brexpiprazole, and lumateperone were approved for schizophrenia treatment, represent a possible therapy to reduce NS. The aim of this rapid review is to summarize the evidence on this topic to make it readily available for psychiatrists treating NS and for further research. We searched the PubMed database for original studies using SDAM, aripiprazole, cariprazine, brexpiprazole, lumateperone, schizophrenia, and NS as keywords. We included four mega-analyses, eight meta-analyses, two post hoc analyses, and 20 clinical trials. Aripiprazole, cariprazine, and brexpiprazole were more effective than placebo in reducing NS. Only six studies compared SDAMs with other classes of antipsychotics, demonstrating a superiority in the treatment of NS mainly for cariprazine. The lack of specific research and various methodological issues, related to the study population and the assessment of NS, may have led to these partial results. Here, we highlight the need to conduct new methodologically robust investigations with head-to-head treatment comparisons and long-term observational studies on homogeneous groups of patients evaluating persistent NS with first- and second-generation scales, namely the Brief Negative Symptom Scale and the Clinical Assessment Interview for Negative Symptoms. This rapid review can expand research on NS therapeutic strategies in schizophrenia, which is fundamental for the long-term improvement of patients\u2019 quality of life. Schizophrenia (SZ) is the 12th leading cause of disability worldwide, with an estimated burden of disease of 13.4 million years of life lived with disability ; p = 0.036). A similar meta-analysis was produced by Zheng et al. (2016) ; p < 0.05) [p < 0.001) [A systematic review and meta-analysis considered a total amount of 25 augmentation strategies for clozapine refractory SZ. Seven placebo-controlled trials considered clozapine augmentation with aripiprazole . NS change was evaluated among the secondary outcomes with the PANSS negative subscale and, where available, with the SANS. Results showed that augmentation with aripiprazole produced improvement in NS in five of the seven RCTs ( < 0.05) . Mitsoni < 0.05) also inv< 0.001) .According to all of the studies cited in this section, as compared to the placebo augmentation arm, the addition of aripiprazole to another antipsychotic, including clozapine, showed greater improvements in NS.The studies on the efficacy of aripiprazole in the treatment of NS are summarized in We included one meta-analysis, one systematic review and meta-analysis, three mega-analyses, five RCTs, and one open-label study that analyzed the efficacy of brexpiprazole in the treatment of NS.Sabe M. et al. (2021) performed a dose\u2013response meta-analysis of forty RCTs that examined the effectiveness of antipsychotics for the acute exacerbation of schizophrenia . In threA systematic review and meta-analysis by Osugo et al. (2022) evaluated the efficacy of dopamine partial agonists and pro-dopaminergic drugs on NS of SZ . BrexpipThree randomized placebo-controlled trials, namely Correll et al. (2015) , Ishigoon = 527) or brexpiprazole 2\u20134 mg/d (n = 878). Some of the subjects enrolled in the brexpiprazole arm during the short-term trials participated in the long-term studies (n = 412 patients). In both short- and long-term studies, patients were stratified into two sub-groups according to symptoms severity. More severely ill were subjects with a PANSS total score > 95 (n = 215), and less severely ill were those with a PANSS total score \u2264 95 (n = 192). In the mega-analysis of the short-term studies, brexpiprazole showed greater improvement than placebo in both sub-groups in both the PANSS Negative subscale and in the PANSS-FSNS [Meade et al. (2020) , conductNSS-FSNS . In the NSS-FSNS .p < 0.01) for the PANSS items emotional withdrawal (N2), poor rapport (N3), lack of spontaneity and flow of conversation (N6), and more significant remission (p < 0.001) in passive/apathetic social withdrawal (N4) and active social avoidance (G16) [Marder et al. (2021) reviewedce (G16) .In conclusion, most of the studies found that brexpiprazole 2\u20134 mg/d was better than placebo in the treatment of NS ,147,148,The studies on the efficacy of brexpiprazole in the treatment of NS are summarized in The studies about the efficacy of cariprazine in the treatment of NS were grouped as follows: cariprazine versus placebo and cariprazine in the treatment of predominant NS and versus other antipsychotics.We found one systematic review and meta-analysis, one mega-analysis, and three RCTs.The efficacy of cariprazine in reducing negative symptoms after 6-week treatment was investigated in three RCTs of patients with an acute exacerbation of SZ . In thesCorponi et al. (2017) chose the three abovementioned RCTs ,153,154 A systematic review and meta-analysis by Osugo et al. (2022) evaluated the efficacy of dopamine partial agonists and pro-dopaminergic drugs on NS of SZ . CaripraIn conclusion, cariprazine seems to be more effective than placebo in the treatment of NS with a dosage between 1.5 mg/d and 6 mg/d.We included one RCT, one open-label study, and one post-hoc analysis.In an open-label, non-controlled study, 60 patients with a confirmed ICD-10 diagnosis of SZ with predominant NS were enrolled . NS weren = 227) or risperidone at the target dosage of 4 mg/d (n = 229). The primary endpoint was the change in PANSS-FSNS. Patients treated with cariprazine, as compared to the risperidone arm, demonstrated significantly greater improvements in the PANSS negative subscale score after week 14 [Nemeth et al. (2017) enrolled week 14 .n = 454 patients). The authors found that cariprazine 4.5 mg/d was significantly more effective than risperidone 4 mg/d in reducing the score of several items included in the PANSS negative subscale [Fleischhacker et al. (2019) set up asubscale . In partsubscale .The three studies summarized in this paragraph excludedIn conclusion, cariprazine was found to be effective and bettThe studies on the efficacy of cariprazine in the treatment of NS are summarized in We found and included the only two available RCTs that considered the effect of lumateperone on NS as compared to placebo.Two RCTs analyzed the efficacy of lumateperone on NS after a 4-week treatment compared with placebo in patients with an acute exacerbation of SZ . The acuLumateperone seems not to be effective for NS in the first four weeks of treatment. At present, long-term studies are not available.The studies on the efficacy of lumateperone in the treatment of NS are summarized in Most of the studies summarized in the present review evidenced that, as compared to placebo, aripiprazole ,136,161,Focusing on the comparison with first- and second-generation antipsychotics, only cariprazine demonstrated evidence of greater efficacy than risperidone in the treatment of NS. Regarding aripiprazole, two studies found a better efficacy of this drug in reducing NS versus amisulpride and rispVarious methodological issues and the lack of studies on the role of SDAMs in the treatment of NS may lead to these fragmented results. In particular, we identified some concerns related to the study population, the experimental design, and the NS assessment. Study populations were heterogeneous within and between studies. Some included only patients with SZ ,159,160,As for the design of the studies, we found a certain degree of heterogeneity. Most of the papers included in the present review were RCTs ,159,160,Finally, many methodological concerns were related to the assessment of NS. Only one study employed second-generation scales in addition to the first-generation ones . This diAnother reason for the lack of evidence of the efficacy of the SDAMs in the treatment of NS was the relatively short period of time that had elapsed following their FDA approval. Possible ways to go beyond these limits are large umbrella trials evaluating different SDAMs and second-generation antipsychotics against a single placebo arm and large, long-term, real-world observational studies. The study samples may include patients clinically stabilized, without treatment-resistant SZ, with a homogenous duration of illness\u2014e.g., maximum 5 years\u2014and with PNS evaluated with first- and second-generation tools. The effect of antipsychotic treatment on the experiential and expressive factors or on each of the five domains of NS should be the primary outcome of the study. A regular and concomitant assessment of positive, depressive, and extrapyramidal symptoms is needed to control for potential secondary NS.The principal limitations of this work are the rapid nature of the review, which did not allow for accurate screening of the methodological quality of the included records, and the choice to search exclusively the PubMed platform and no other databases such as Web of Science.The main strength of this paper is the choice of the topic covered, previously not addressed by other reviews. Indeed, this rapid review represents the first synthesis of the evidence on the efficacy of the four SDAMs approved for the treatment of NS in SZ.The main implications of the findings of this review concern the use of cariprazine and lumateperone to treat NS. In particular, cariprazine was the only SDAM that demonstrated, in large studies with sound methodology, a superiority to an SGA while lumateperone, contrary to the other three SDAMs, did not show superiority to placebo. For this drug, further studies with longer follow-ups are needed. Moreover, the current rapid review highlights the need for new, methodologically sound studies aimed at demonstrating the efficacy of SDAMs in the reduction of NS. This goal may be achieved by selecting samples of patients with SZ homogenous in terms of duration of illness, with a detailed characterization of persistent NS assessed with first- and second-generation scales and grouped in the experiential and expressive factors. Three study designs may be suitable for this purpose, namely long-term and significantly large sample RCTs to determine the duration of treatment and the maintenance of antipsychotic doses , head-toIn conclusion, the SDAMs represent a possible option for the treatment of NS in SZ. However, the evidence on the efficacy of these drugs is still poor and fragmented because of the lack of studies on this topic and of several methodological issues concerning the choice of the sample, the evaluation of NS, the study design, the follow-up duration, and the comparators, mainly placebo and not another antipsychotic. Only a few studies, mainly on cariprazine in monotherapy, confirmed the superiority of this group of drugs to other classes of antipsychotics. Therefore, according to the available evidence synthesized in the current review, among SDAMs and antipsychotics in general, only cariprazine may represent an effective strategy to reduce these disabling symptoms. New, methodologically robust studies focusing on well-characterized patients with head-to-head comparisons of antipsychotic treatments and long-term real-world observational studies are needed to fill this knowledge gap."} +{"text": "Mizaj is an individualized viewpoint in Persian Medicine (PM) that is used for the prevetion of diseases and also treatment. Evaluating Mizaj in the two domains of hotness-coldness, and wetness-dryness, 10 criteria have been introduced, most of them are qualitative. To achieve valid and reliable questionnaires, the weight of these criteria must be determined in assessing the Mizaj. In a cross-sectional study with Delphi method, 10 indices were extracted from PM references and sent to PM experts via e-mail. They were asked to score the weight of each index in determining the Mizaj from 0 to 10. The scores ranked and comparing previous preliminary studies, criteria of major and minor were proposed.Out of 147 invited PM experts, 122 completed the tables. Based on scores, physical functions, physique, and responsiveness of organs obtained the highest scores in the field of hotness-coldness. In wetness-dryness muscle/fat mass and sleep/wakefulness received the highest scores from the viewpoint of experts.Physical functions, physique (Anthropometry), responsiveness of organs and psychic function can be used as major criteria in Mizaj assessment methods in the hotness-coldness field. In the field of wetness-dryness, muscle/fat mass, sleep/wakefulness, tactile condition and physique (anthropometry) can be considered as major criteria. A worldwide trend to complementary and traditional medicine recently has been on the rise . Differe These include tactile condition (the condition of the skin of the body from the examiner's point of view), muscle/fat mass , hair condition , skin color (various colors of skin), physique , responsiveness of organs , sleep/wakefulness , physical functions , body waste , and psychic function . Based oThis study is a cross-sectional study that was done from September 2020 to February 2021 at the Research Center of Traditional Medicine and History of Medical Sciences in Babol University of Medical Sciences, Iran. It was approved by the Ethics Committee of Babol University of Medical Sciences. This study was conducted with Delphi method. The Delphi method is a structured process for collecting and classifying the knowledge available to a group of experts , 20. In this study, based on the main PM references 10 criteria for determining whole body Mizaj were extracted of the participants were females and 44 (36%) were males. The mean duration of their experience in practice of PM was 10.67 \u00b15.45 years. As a result, physical functions, physique, and responsiveness of organs had the highest scores in the field of hotness-coldness. In wetness-dryness muscle/fat mass and sleep/wakefulness received the highest scores from the experts. The details of scores are shown in 9. Giving weight to the proposed indices are essential to develope new valid and reliable questionnaires and other diagnostic tools. In this study, Delphi method as a structured process was used , responsiveness of organs and psychic function can be used as major criteria in Mizaj assessment methods in the hotness-coldness field. Moreover, in the field of wetness-dryness, muscle/fat mass, sleep/wakefulness, tactile condition and physique (anthropometry) can be considered as major criteria."} +{"text": "Magnetic Resonance Imaging and Spectroscopy have become indispensable diagnostic tools in numerous medical applications, providing anatomical, functional, and chemical information non-invasively with ever increasing sensitivity and specificity. However, many performance challenges are present in connection with improving the sensitivity, safety, and accessibility of these modalities. In this context, a traditional strategy to improve sensitivity is to develop ultra-high field scanners, but this gives rise to numerous technical hurdles, physics related concerns, and safety issues in addition to increased exclusiveness. More recently, the development of ultra-low field MR systems has also resurfaced in an effort to focus on accessibility, inherently, avoiding safety issues and wave propagation-related image distortions most typical of high fields. However, their inherently low MR signal amplitude comes at the cost of poor sensitivity.Both the ultra-high and ultra-low field domains, consequently, call for scientific innovations that pave the way for the next generation of MR hardware systems. Today\u2019s challenges in the development of MR instrumentation arise from the different requirements and limitations faced at different magnetic fields strengths, and all innovations target the improvement of image quality, safety, and/or diagnostic value of MR.Consequently, this Research Topic has invited submissions on all innovative developments in MR hardware over the entire range of static magnetic field strengths. The resulting collection of articles thus covers a large variety of topics, ranging from ultra-low to ultra-high field MR, including intermediate field strengths, spanning work on radio frequency components, magnet and gradient design, as well as complete MR systems. The Research Topic consists of mostly Original Research articles, augmented by a Perspective and a Mini Review article, as well as a paper in the category Technology and Code.Wenz and Gruetter investigate the impact of quasi-transverse electric modes on the transmit field distribution using dipole-fed rectangular dielectric resonator antennas at 7 T. Also, Williams et al. propose a nested 8-channel transmit array with an open-face concept for human brain imaging at 7 T. In her Mini Review article, Irena Zivkovic discusses available interelement decoupling strategies for ultra-high field MR coils. Van Leeuwen et al. discuss a potential reduction in peripheral local SAR for a birdcage body coil at 3 T using a magnetic shield.At ultra-high field strengths, the strongest unmet need concerns the improvement of radiofrequency (RF) transmission systems regarding homogeneity of the RF excitation, efficiency, decoupling strategies, and patient safety with respect to the associated specific absorption rate (SAR) limitations. In this sense, 1H), the most challenging task is to increase the sensitivity of the RF detection system, driven by the need to overcome the sensitivity-based limitation of image quality and spatio-temporal resolution set by the various noise sources present in the MR experiment. To this end, several strategies employing new coil geometries, new materials, or cryogenic devices are pursued.At ultra-low field, but also for MR of X-nuclei (i.e., other than Labb\u00e9 et al. present their results on high-temperature superconducting RF coils, exploiting the intrinsically low coil noise of such devices. New RF coil shapes are reported by Nowikow et al. in their contribution about Koch Snowflake fractal geometry coils for 23Na MRI.The investigation of novel RF coil element types is an active domain of hardware development for MRI, with the ultimate goal of improving sensitivity. Harper et al. in their work about an unmatched RF chain for low field MRI. In addition, the trend towards portability and ease of use of MR instrumentation calls for other features that increase usability and accessibility but imposes novel technological challenges, on the one hand regarding magnet design for portable MRI systems, and on the other hand flexible, lightweight or wireless RF devices. De Vos et al. introduce an improved portable and sustainable low-field MRI system, complemented by B0 shimming methodology for affordable and compact low-field MRI magnets as described by Wenzel et al. In a computational and phantom study at ultra-low field MRI, H\u00f6fner et al. investigate the feasibility of 3D neuronal current imaging, relying on the measurement of the minuscule phase perturbation generated by the neuronal magnetic fields.The need for innovative electronics for ultra-low field MR is evidenced by Littin et al. present an intuitive open-source collection that can be employed to find new gradient coil designs. In an endeavor to reduce acoustic noise and heat in gradient coils, Motovilova and Winkler review existing techniques and present solutions for quiet operation and efficient gradient cooling.Regardless of field strength, the development of improved gradient designs enables novel applications of MRI and increases imaging performance. In their Technology and Code article, Galuppini et al. discuss the key challenges of field-frequency lock in fast field cycling MRI and define possible research directions in the field.Finally, in a Perspective article, We, the editors of this Research Topic, hope that the readers will derive added value from the presented collection of articles. We are convinced that innovative hardware concepts and technological development, such as the works presented here, will be highly beneficial for MR research at all field strengths in the future. By combining papers on seemingly very disparate subtopics in MR hardware development into a single collection, we aim at establishing stronger links within the community, so as to foster unconventional and creative solutions for the upcoming challenges in the development and improvement of MR systems."} +{"text": "In the published article, there was an error in the Funding statement. The grant number included was incorrect. The original text incorrectly states that \u201cThis work was supported by Medical Research Centre (Grant No. RP # 16354/16), Hamad Medical Corporation, Doha, Qatar. The publication of this article was funded by the Qatar National Library\u201d. The correct Funding statement appears below.FUNDINGThis work was supported by Medical Research Centre (Grant No. MRC-01-18-186), Hamad Medical Corporation, Doha, Qatar. The publication of this article was funded by the Qatar National Library.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way."} +{"text": "Some impediments in language learning may have a detrimental impact on learners\u2019 actual performance on the test and lead to anxiety and demotivation. Language achievement is influenced by self-assessment (SA), academic resilience (AR), academic motivation (AM), and test-taking skills (T-TS) among other factors. Considering the relevance of these factors in language achievement, the current investigation aims to delve into the probable interactions of SA, AR, AM, T-TS, and test anxiety (TA) management among English as a foreign language (EFL) learners. A model was devised and evaluated using confirmatory factor analysis (CFA) and structural equation modeling (SEM) to achieve this objective. This research collected 512 by distributing online questionnaires to fifteen approved private institutions which applied Telegram-based language learning. The study findings reflected that SA, AR, and AM could predict EFL learners\u2019 T-TS. It was also confirmed that SA, AR, and AM modulated EFL learners\u2019 TA. The implications of the study are presented and accompanied by some future research proposals as well as instructional consequences. Several factors impact the decisions that educators and test developers must make to establish and administer an effective education and assessment program. Both emotive and cognitive characteristics of the learners are critical in successful language testing. Practically speaking, learning-oriented evaluation in the classroom operates reasonably well to evaluate language learners\u2019 language progress and observe their mental and emotional well-being in terms of their T-TS and TA, this research sought to explore the contribution of SA, AM, AR to T-TS, and TA management. Holding these standpoints into consideration led to the formulation of the following research questions:H01: EFL learners\u2019 SA does not provide insight into their T-TS in Telegram-assisted-language learning.H02: EFL learners\u2019 SA does not provide insight into their TA in Telegram-assisted-language learningH03: EFL learners\u2019 AR does not provide insight into their T-TS in Telegram-assisted-language learning.H04: EFL learners\u2019 AR does not provide insight into their TA in Telegram-assisted-language learningH05: EFL learners\u2019 AM does not provide insight into their T-TS in Telegram-assisted-language learningH06: EFL learners\u2019 AM does not provide insight into their TA in Telegram-assisted-language learningIn this respect, the following null hypothesis are proposed:As a response to these inherent gaps, a model was developed to illustrate the probable linkages between SA, AR, AM, T-TS, and TA. This model was developed based on pertinent theories and contemporary literature see Fig.\u00a0. Using CThe term \u201cassessment\u201d encompasses an array of methods that are used to analyze and draw conclusions about the advancement that pupils have made in their studies Bachman, . Many alResilience has been conceptualized as the ability to sustain normal development and make positive adaptations while facing significant adversity , which was first proposed by Deci and Ryan , is now TA may be experienced by learners before, during, and after the test. Horwitz et al. introducThe COVID-19 pandemic has interrupted education in many countries; thus, as an emergent reaction, several governments have prioritized the implementation of virtual instructional strategies. During the COVID-19 outbreak, online courses and social media facilitated two-way communication between pupils and their instructors Peng, . In receTelegram is compatible with the operating systems used by Android, iOS, and Windows Phone, as well as Mac and Windows devices. Furthermore, the Telegram app may be used concurrently from a number of different devices . Among the subcomponents of AR, Perceived Happiness received the highest mean score. Regarding AM, the mean score was . Furthermore, During-Test was the highest among T-TS. In the final variable, TA, Incapacity was different . Then, the normality of the data was assessed and reported in Table As Table Based on what Table \u03b2\u2009=\u20090.84, t\u2009=\u200929.54) and TA was positive. The effect of AR on T-TS and TA was significant. The same was true about the impact of AM on T-TS and TA .The causal relationships among the variables are portrayed in Figs.\u00a0The data in Table \u03b2\u2009=\u20090.89, t\u2009=\u200929.67), Time Management , During-Test , After-Test , Incapacity , and Worry and Dread . The association between AR, TS, and TA subfactors is as follows: AR and Before-Test , Time Management , During-Test , After-Test , Incapacity , and Worry and Dread . Additionally, AM is linked with TS, and TA subfactors are as follows: Before-Test , Time Management , During-Test , After-Test , Incapacity , and Worry and Dread .In model 2 which is illustrated via Figs.\u00a0As the final step, a Pearson product-moment correlation was run to address the correlation between SA, AR, AM, T-TS, and TA subfactors.Table This research intended to investigate the impact of the SA, AR, and AM on T-TS and TA management in academic settings. To accomplish this goal, an empirical study was carried out with EFL students enrolled in English language institutions learning English via Telegram. The analysis of the data revealed that enhancement in SA, AR, and AM enables learners to pass their tests skillfully and manage their possible test anxiety. These findings underlined the crucial part that social media, specifically the Telegram app, plays in improving students\u2019 mental and emotional health, and the suggested model Fig.\u00a0 was confTaking into consideration the first and the second research questions, which were \u201cRQ1:Can EFL learners\u2019 SA provide insight into their T-TS in online classes? and RQ2: Can EFL learners\u2019 SA provide insight on their TA in online classes?\u201d, the findings of this exploration show that SA can play a moderator function in T-TS and TA management (model 1 & model 2). It is possible to infer that how students approach self-evaluation may help them build or restore a positive self-image and a strong sense of self-worth, which in turn helps to reinforce the accomplishment of language learning objectives. Furthermore, SA helps learners cultivate an internalized confidence in their abilities and a perception of effectiveness in their skills to complete assignments effectively . Based on the opportunities provided by the Telegram app, learners can easily communicate, and the anxiety of being judged by their peers would decrease. This outcome is in accordance with Esmailzade Ashini et al. , who conThe results of the fifth and sixth research questions revealed that AM played a significant role in increasing EFL learners\u2019 T-TK and decreasing TA. That is, AM inspires students to act with vigor and build a good self-image and gives them assistance and engagement (model 1 & model 2). Both D\u00f6rnyei et al. and Al-HIn a nutshell, the purpose of this inquiry was to uncover the connection between SA, AR, AM, T-TS, and TA management in the EFL context. In this respect, a model was hypothesized and then verified using CFA and SEM in the proper manner. Furthermore, this study was to provide the spotlight on the advantages that Telegram-based training may bestow to language assessment when applied in an EFL environment. The obtained data provide support for the proposed model and reflect the predictive capacity of SA, AR, and AM, respectively.In light of the results of the research, there are a number of pedagogical recommendations that may be made. Implementation of MALL into the curriculum has the potential to boost and extend teachers\u2019 and student access to course data and foster LOA whenever and wherever, even outside of the classroom, as the outcome of the swift development of emerging technologies and the need for hosting online and virtual courses. This enhancement is of great importance in the realm of language learning and assessment. As it was displayed in this the current research, language learners felt more secure and less anxious. As such, educators need to learn the digital literacy necessary to use MALL platforms, notably the Telegram app, for the purpose of language instruction and assessment. It is necessary for both instructors and language learners to acquire an awareness of the self-aid conceptions and the attributions such constructs have on their well-being and LOA enhancement. It is important for the academic courses to either directly or indirectly include training in the relevant methods.It is recommended that those responsible for developing academic curricula and materials make changes to such materials and take these results into consideration when working on other academic fields. It is advised that policymakers, curriculum designers, material producers, test developers, and language teachers give some thought to language instruction based on MALL and teaching self-aid practices. This will ensure the academic success of the learners, LOA, and, more significantly, the well-being of society. In addition, practical tactics that may be used to cultivate and put into practice SA, AR, AM, and T-TS should be introduced into higher education. Language teachers may get the necessary training via pre-service and in-service training courses, respectively.Although this research provides interesting insights into the topic in question, it faces some limitations, which need to be addressed. To get things started, all of the learners who participated in our investigation were aged between 17 and 23 with similar English proficiency. In forthcoming studies, it may be possible to evaluate the correlations between SA, AR, AM, T-TS, and TA management in a variety of educational environments and among students majoring in a variety of different fields of study. In addition, due to the fact that all of the information gleaned from this study was obtained via the self-reporting of the learners on questionnaires, the generalizability of the findings is under question. In future research, incorporating qualitative and quantitative methods may provide more thorough findings. This study was limited to telegram-assisted language learning. As a future research avenue, it is suggested to investigate the effects of applying other apps in language learning enhancement and psychological health of the learners. Moreover, an additional survey is suggested to triangulate the study findings and to undertake a comprehensive analysis of the degree to which demographic factors of the students may have an impact on the interrelationship among SA, AR, AM, T-TS, and TA management."} +{"text": "Physical activity and sedentary behaviour have received intense interest in recent decades from a variety of stakeholders due to their diverse association with cognitive function, which is a key component in maintaining health of older adults. The aim of this bibliometric analysis is to reveal the developmental structure, research hotspots, and predict future trends in the field of physical activity, sedentary behaviour, and cognitive function research among older adults.This study adhered strictly to the step-by-step guideline of bibliometric analysis. Publications in this field from 2002 to 2022 were retrieved from the Web of Science Core Collection. Only original articles and review articles published in English were included in the analysis. A cubic polynomial was applied to determine the publication trends. CiteSpace (6.1.R4 Basic) and VOSviewer (1.6.18) were performed to obtain collaborative networks, reference co-cited networks, and keyword co-occurrence networks, as well as to perform burst analysis to predict future trends.A total of 1093 publications were retrieved, of which 73.56% obtained research grants. The publications were cited 30,317 times. Within two decades there was rapid growth in publications in this field. The USA and China were the top contributors, and the USA had the strongest collaborative network. University of British Columbia and University of California System were the most active institutions. Liu-Ambrose T and Shimada H contributed to the largest number of publications. Journal of Aging and Physical Activity, and Frontiers in Aging Neuroscience were the most active journals. Reference co-cited networks, and keyword co-occurrence networks revealed that hotspots in this field were age, nursing home resident, tai chi, and perceptions, while future trends in this field were healthy ageing, qi gong, sedentary behaviour, and falls.There has been an increased research interest in the interrelationships between physical activity, sedentary behaviour, and cognitive function in older adults in the past two decades. Our findings will help researchers, practitioners, and other stakeholders to understand the structure and hotspots in this relevant field of research.Not applicable."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Social justice design and implementation in library and information science. New York: Routledge; 2021. Softcover: 332p. ISBN 978-0-367-65382-8. Price: USD$48.95. Available from: https://www.routledge.com/Social-Justice-Design-and-Implementation-in-Library-and-Information-Science/Mehra/p/book/9780367653828Mehra B, editor. Social justice design and implementation in library and information science presents a diversity of cases pertinent to librarianship, information science research, and library and information science (LIS) education. Editor Dr. Bharat Mehra is the EBSCO Endowed Chair in Social Justice and Professor in the School of Library & Information Studies at the University of Alabama. Dr. Mehra earned his PhD in Library and Information Science from the University of Illinois at Urbana-Champaign after also earning degrees in South Asian and Middle Eastern Studies and Landscape Architecture. This background sheds light on the selection of topics in this collection, including a chapter discussing cultural heritage studies as applied to built spaces (chapter 9).I volunteered to review this book because of my interest in the topic. I hoped the text would help me generate ideas for my workplace\u2019s Diversity, Equity, and Inclusion Committee and Hiring Practices Evaluation Taskforce. As an instruction librarian and liaison to pre-clinical medical school courses, I was interested to see how social justice discussions in health professional education compared to similarly focused conversations in my own professional community. I acknowledge my own bias for cases in settings most relevant to my own workplace.The case studies in this text vary greatly, not only in terms of topics and settings, but also in their presentation and seemingly intended audiences. Cases are organized into five sections: (I) emerging new responsibilities, (II) reflective case practices, (III) reaching out: new research approaches and strategies, (IV) transforming LIS education, and (V) instruments of action and change. These categories try to cover the broad range of questions relevant to social justice.Notably, authors take a variety of approaches in presenting their work. Some case studies follow a traditional structure: background, literature review, and statement of purpose. Reflecting a familiar outline, these chapters appealed to me most. Chapter 4, for instance, presents a structured case study of a public library involved in food justice. Though the setting and the intervention (food justice) differ greatly from my own work, I was able to relate to themes described by the authors. Not all chapters provide such entr\u00e9e points.Other cases are grounded in theory. While interested in the theory of our profession, I found these chapters less practical and relevant to my role as a reference and instruction librarian in an academic medical center. They may be more pertinent to the work of information science researchers and LIS educators.Finally, some contributors take a narrative approach. This documentation is crucial as these stories are often lost. In the foreword, Paul T. Jaeger, Co-Director of the Information Policy and Access Center (iPAC) and former MLIS program Co-Director at the College of Information Studies, University of Maryland, describes the importance of the current collection in documenting our profession\u2019s legacy.Our professional roles and research interests are broad and their intersections with social justice are many, as demonstrated in the diversity of content within this text. I appreciated learning about the work of librarians and information professionals in this space, but I wanted more tie-ins to larger contexts, descriptions of impact, lessons learned, or most useful for me, practical tips and potential applications to other settings.Future printings should correct typos and copyediting mistakes. I was often distracted from the content by these errors. Additionally, I found it difficult to switch between chapters, as some used dense, jargon-laden text, whereas others used plain language and storytelling style.Overall, I found the stories illuminating and inspiring. I was able to relate in some ways, even to librarians working in other contexts. Personally, I am interested in seeing how professional schools adapt and update curricula to meet the ever-changing needs of the workplace, as described by cases in section IV, transforming LIS education. With its expansive view of social justice in library and information science, this text would be a good addition to collections serving library and information science schools or degree programs. Yet, even while I acknowledge the importance of its topic, I would not recommend this book for purchase by a health sciences library."} +{"text": "Psychiatryai.com is a prototype Artificial Intelligence (AI) and Data Science (DS) platform and research project developed for my Evidence-Based Healthcare (EBHC) course at University of Oxford in MSc studies (Kellogg College). This is a singular, multi-disciplinary, and beta-testing project in Computing Science, Psychiatry, and Mental Health for oral presentation at EPA 2023.near real-time about psychiatry and mental health evidence-based research - for healthcare professionals, doctors, and researchers in psychiatry. The project also aims to integrate findings from the Goldacre Review (2022) into practice and develop novel computing solutions utilising AI and DS, and present findings.AI and DS in Psychiatry and Mental Health have emerged as important research areas in the post Covid-19 pandemic era. This prototype University project (Psychiatryai.com) was launched on 22nd November 2021. It aims to develop a free, secure, and open access platform in A WordPress site (Psychiatryai.com) was developed with syndicated RSS feeds across 330 psychiatry topics and refreshed by data servers hourly, 24 hours a day, 7 days a week. A total of 43 WordPress plugins were utilised to develop this secure platform. The site is powered by intuitive data modelling and analytics in near real-time and available in open access coding format for peer-review, future development, and research. The primary sources of live evidence for the project are PubMed and University of Helsinki, Finland. The server performance data analytics will be available for poster presentation at EPA 2023. This includes full statistical results and discussion since its inception and launch , and robust technical analysis and performance outcomes - available freely online to promote research in psychiatry and mental health.Knowledge Synthesis and Dissemination:Total Words: 4356886 *Live Psychiatry and Mental Health Citations from PubMed: Exceeds 325000Total Evidence Alerts Published: 54391 *Total Algorithms/Topics: 330Total site visitors: 8023 ** Since launch of Psychiatryai.com on 22 November 2021 inclusive to 31 October 2022full real-time data analytics and dissemination of peer-reviewed current evidence in the future. The emergence of these technologies will be useful in settings such as disaster psychiatry, psychiatry e-training and research, and e-mental health awareness/promotion ahead.Psychiatryai.com was able to demonstrate succesful development of an effective and viable platform to study AI and DS in Psychiatry and Mental Health, as evidenced by results table. The platform has also incorporated findings from the Goldacre Review (2022) and aims to continue to collect valuable insights towards P. Naik Consultant of: Dr Paras Naik (Psychiatryai.com) Non-profit University Project"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.1As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.3Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.5Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.7Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.16Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.22Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.10In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "In January, 2023, the Science and Security Board of the Bulletin of the Atomic Scientists moved the hands of the Doomsday Clock forward to 90s before midnight, reflecting the growing risk of nuclear war.Current nuclear arms control and non-proliferation efforts are inadequate to protect the world\u2019s population against the threat of nuclear war by design, error, or miscalculation. The Treaty on the Non-Proliferation of Nuclear Weapons (NPT) commits each of the 190 participating nations \u201cto pursue negotiations in good faith on effective measures relating to cessation of the nuclear arms race at an early date and to nuclear disarmament, and on a treaty on general and complete disarmament under strict and effective international control\u201d.Any use of nuclear weapons would be catastrophic for humanity. Even a \u201climited\u201d nuclear war involving only 250 of the 13 000 nuclear weapons in the world could kill 120 million people outright and cause global climate disruption leading to a nuclear famine, putting two billion people at risk.The health community has had a crucial role in efforts to reduce the risk of nuclear war and must continue to do so in the future.In 2007, the IPPNW launched the International Campaign to Abolish Nuclear Weapons, which grew into a global civil society campaign with hundreds of partner organisations. A pathway to nuclear abolition was created with the adoption of the Treaty on the Prohibition of Nuclear Weapons in 2017, for which the International Campaign to Abolish Nuclear Weapons was awarded the 2017 Nobel Peace Prize. International medical organisations, including the International Committee of the Red Cross, the IPPNW, the World Medical Association, the World Federation of Public Health Associations, and the International Council of Nurses, had key roles in the process leading up to the negotiations, and in the negotiations themselves, presenting the scientific evidence about the catastrophic health and environmental consequences of nuclear weapons and nuclear war. They continued this important collaboration during the First Meeting of the States Parties to the Treaty on the Prohibition of Nuclear Weapons, which currently has 92 signatories, including 68 member states.11We now call on health professional associations to inform their members worldwide about the threat to human survival and to join with the IPPNW to support efforts to reduce the near-term risks of nuclear war, including three immediate steps on the part of nuclear-armed states and their allies: first, adopt a no first use policy;The danger is great and growing. The nuclear armed states must eliminate their nuclear arsenals before they eliminate us. The health community played a decisive part during the Cold War and more recently in the development of the Treaty on the Prohibition of Nuclear Weapons. We must take up this challenge again as an urgent priority, working with renewed energy to reduce the risks of nuclear war and to eliminate nuclear weapons.Kamran Abbasi, Editor-in-Chief, British Medical Journal;Parveen Ali, Editor-in-Chief, International Nursing Review;Virginia Barbour, Editor-in-Chief, Medical Journal of Australia;Kirsten Bibbins-Domingo, Editor-in-Chief, Journal of the American Medical Association;Marcel GM Olde Rikkert, Editor-in-Chief, Dutch Journal of Medicine;Peng Gong, Editor-in-Chief, Chinese Science Bulletin;Andy Haines, London School of Hygiene and Tropical Medicine;Ira Helfand, Past President, International Physicians for the Prevention of Nuclear War;Richard Horton, Editor-in-Chief, The Lancet;Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine;Arun Mitra, Past President, International Physicians for the Prevention of Nuclear War;Carlos Monteiro, Editor-in-Chief, Revista de Sa\u00fade P\u00fablica;Elena N. Naumova, Editor-in-Chief, Journal of Public Health Policy;Eric J. Rubin, Editor-in-Chief, New England Journal of Medicine;Tilman Ruff, Past President, International Physicians for the Prevention of Nuclear War;Peush Sahni, Editor-in-Chief, National Medical Journal of India;James Tumwine, Editor-in-Chief, African Health Sciences;Paul Yonga, Editor-in-Chief, East African Medical Journal;Chris Zielinski, University of Winchester, World Association of Medical Editors.This editorial is being published simultaneously in multiple journals. For the full list of journals see: https://www.bmj.com/content/full-list-authors-and-signatories-nuclear-risk-editorial-august-2023"} +{"text": "Correction to: Clinical and Translational Oncology 10.1007/s12094-023-03216-3In Table The Acknowledgments and Conflict of interest sections were missing from this article and should have read as follows.AcknowledgmentsThe authors thank Enriqueta Felip y Javier de Castro for their review and validation of the levels of evidence and grades of recommendation in this guideline.Conflict of interestRGC reports Advisory boards, Consultancy and Speaker honoraria from MSD, BMS, Roche, Boehringer Ingelheim, Pfizer, Novartis, AstraZeneca, Lilly, Takea and Amgen. IS reports Advisory Board from Roche, Novartis, Boehringer Ingelheim, Takeda and Sanofi; Speaker from Roche, MSD, Pfizer, BMS and AstraZeneca; Grant from Roche, Takeda, Pfizer, BMS and AstraZeneca; Non-financial Support from Member of GECP. EA reports Advisory Board, Speakerand Grant from Roche; Advisory Board and Speaker from Astra Zeneca and BMS; Advisory Board, Speaker and Non-financial Support from Takeda; Advisory Board from Lilly and Boehringer-Ingelheim; Speaker from MSD, Merck, Thermo Fisher Scientific and Guardant Health; Speaker and- Non-financial Support from Pfizer. AI reports Advisory Board and Speaker from Roche, AstraZeneca and Sanofi; Speaker from Takeda and BMS. OJJV reports Advisory Board and Speaker from BMS and Janssen; Speker from Roche; Advisory Board, Speaker and Other from AstraZeneca, Takeda and Janssen; Advisory Board from Lilly. NR reports Advisory Board, Speaker and- Other from MSD; Advisory Board and Speaker from AstraZeneca, Takeda, Amgen, Lilly, Sanofi, Roche and Janssen. JZ reports Speaker, Grant and Personal Feels from BMS and AstraZencea; Advisory Board and Personal Feels from Sanofi; Speaker from MSD; Grant and Personal Feels from Roche; Speaker and Personal Feels from Pfizer; Advisory Board from Novartis and Speaker from NanoString. MD reports Advisory Board and Speaker from AatraZeneca, Pfizer and Takeda; Speaker from BMS, MSD and Roche; Advisory Board from Janssen and Sanofi. TM and PCC have nothing to disclose.The original article has been corrected."} +{"text": "Correction: Genetics Selection Evolution (2022) 54:66 https://doi.org/10.1186/s12711-022-00752-4After publication of this work , we notiThe correct equations should be:Then,Therefore,Please note that the equations are correctly programmed in the software used for analyses, and the corrections above do not affect the end results presented in the manuscript.We thank Dr. Matias Berman from The University of Georgia and Dr. Jeremie Vandenplas from Wageningen University for their interest and suggestions to correct the manuscript."} +{"text": "The field of soft matter teems with molecules and aggregates of molecules that have internal size-modulating degrees of freedom. Proteins, peptides, microgels, polymers, micelles, and even some colloids can exist in multiple\u2014often just two dominating\u2014states with different effective sizes, where size can refer to the volume or to the cross-sectional area for particles residing on surfaces. The size-dependence of their accessible states renders the behavior of these particles pressure-sensitive. The Bragg\u2013Williams model is among the most simple mean-field methods to translate the presence of inter-particle interactions into an approximate phase diagram. Here, we extend the Bragg\u2013Williams model to account for the presence of particles that are immersed in a solvent and exist in two distinct states, one occupying a smaller and the other one a larger size. The basis of the extension is a lattice\u2013sublattice approximation that we use to host the two size-differing states. Our model includes particle\u2013solvent interactions that act as an effective surface tension between particles and solvent and are ignorant of the state in which the particles reside. We analyze how the energetic preference of the particles for one or the other state affects the phase diagrams. The possibility of a single phase-two phases-single phase sequence of phase transitions as a function of increasing temperature is demonstrated. The level of detail for modeling the phase behavior of particle systems ranges from atomistic descriptions ,2,3,4 toAmong the systems that have their properties affected by internal degrees of freedom are hydrogels ,9, elongOn the level of mean-field theory, we have previously presented a latticS\u201d and \u201cL\u201d to denote quantities related to particles in the small-volume and in the large-volume state, respectively. The numbers of particles in each state, M equal sites, it is convenient to introduce the corresponding volume fractions S particle and a solvent molecule occupy one single lattice site each. More general cases with different volumes for solvent molecules and S particles are straightforward to implement. S, light red cubes) and large and (b)\u2014to two coexisting phases, colored in white and blue, and each with distinct compositions and phase size\u2014diagrams (c) and (d). By further increasing L particles. Finally, for even larger S particles. The color gradient in the arrows indicates the increase in We consider a binary mixture of solvent molecules and a fixed number of constant ; while NL-state and S-state, T and the configurational entropy by S. A lattice\u2013sublattice approximation proposed by Han and co-workers [L-state to the S-state. Finally, the particle\u2013solvent interactions, per lattice site, are accounted for on the mean-field level by the expressionS-state over the L-state of isolated particles that are surrounded by solvent. Note that both S-state and L-state in the same way, and S particles and L particles do not interact with each other. In a more general approach, one would assign distinct interaction strengths to pairs of L particles and solvent, S particles and solvent, and L particles and S particles. With the energy contributions specified above, the (dimensionless) free energy per lattice site, The Helmholtz free energy of the system, -workers allows upressionUintMkBT=S-state and L-state, we must first obtain the equilibrium distributions. To this end, we insert the relation L particles versus S particles, readsTo analyze how the interaction strengths Equation and findKnown truction .We start our discussion by presenting in Upon increasing global minimum. The main diagrams magnify the region The results shown in Equation , thus beS-state or L-state; recall that L) to its small (S) size [The limit of small (S) size and thatL-state or S-state. When S-state dominating. Growing L-state is energetically preferred over the S-state, this may lead to a transition to a dense phase dominated by the L-state. Let us analyze which state, L or S, dominates in the limit of small s:Upon increasing Equation and expaL-state (bottom left inset), and another, for large s, where virtually all particles are in the S-state (top right inset). We note that the larger S-state. Furthermore, if L-state. This trend, however, does not continue when L-state are placed on the lattice until steric constraints force them to undergo a transition to the smaller S-state.Results from Equation are plotEquation , Figure T because In s common ,45,46, wlculated ,48 from As our parameter space has five dimensions , the critical point is L-state is enforced, the critical point is located at In order to investigate the effects of further increasing S-state is enforced. Hence, as in this case the system consists exclusively of particles in the S-state, all lines converge to the Bragg\u2013Williams result irrespective of the value of L-state. Consequently, the system behaves as composed solely of particles in the L-state, with particle volume The three sets of dashed lines, on which the corresponding symbols lie, were calculated for P of the system throughWe finally note that the function alculate the presP at fixed P, despite the growing preference for the L-state. The pressure tends to increase with In L) and another of a smaller size (S). Our model is based on a lattice\u2013sublattice approximation [L-state to the S-state, and L-state and S-state. In the limit of very large positive or negative Molecules with internal degrees of freedom are ubiquitous in nature. Colloids, proteins, and microgels, are just a few examples of molecules that can adjust their size in response to external stimuli. In the present work, we developed a simple extension of the Bragg\u2013Williams model to describe solvent-immersed particles that can exist in two states, one of a larger (ximation ,43 and asotherms ,39,40 ansotherms ,50."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.1As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.3Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.5Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.7Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.16Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.22Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.10In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "This study aimed to reveal the epidemiological trends of CL in the Tabuk region, the Kingdom of Saudi Arabia (KSA), during the period from 2006 to 2021.patients with CL, who were detected and registered at the regional Vector-borne Diseases Control Unit of the Tabuk province, between January 2006 and December 2021, were analyzed in this retrospective study. The patients\u00b4 data included their nationality, gender, and age, and their annual and month-by-month recorded patterns.a total of 1575 CL patients were reported during the said period. They were 53.1% Saudis and 46.9% non-Saudi expatriates with a ratio around 1.1: 1.0; and they were re-categorized as 83.17% males and 16.83% females with a ratio of 4.9: 1.0 (p <0.5). Additionally, the majority of these CL patients were in age group of 15-45 years (p <0.5), and the lowest number was in age group of <5 years. Most importantly, there was a continuous annual and month-by-month record of these patients; reflecting CL endemicity in the Tabuk region of KSA.the present findings imply that CL is endemic in the Tabuk region of KSA. As there is a recent increase in human immigration to this region, sustainable monitoring of CL and improving its control measures is warranted. Leishmania. There are more than twenty different Leishmania species known to infect humans, and their transmission is carried out by the bite of the infected female phlebotomine sandflies [Leishmaniasis is a complex vector-borne disease caused by an obligate intracellular flagellated protozoan parasite of the family Trypanosomatidae, order Kinetoplastida, and genus andflies ,2. Epideandflies ,4. Annuaandflies ,6, and uandflies .Leishmania species can infect all human ages and present multiple manifestations [Clinically, stations . Dependistations -11. Globstations ,12-14. Mstations . As a costations ,17. At tstations and sincstations -21. The Study design and setting: the present retrospective study was designed to reveal the epidemiological trends related to CL infection among individuals who were resident in the Tabuk region, KSA, during the period from January 2006 to December 2021. An ethical approval (IRB # TU-077-022-127) was obtained from the Institutional Review Board of Health Affairs in the Tabuk province, and patients with confirmed CL during the said period, and they were registered at the Vector-borne Diseases Control Unit of the Tabuk province, were included here. Following the guidelines of the established vector-borne diseases control program by the Saudi Ministry of Health (MoH), the diagnosis and treatment of any vector-borne positive case, including leishmaniasis, is totally hospital-based, and case information are directly delivered to the Vector-borne Diseases Control Unit of this region through a specific health program axis. Geographically, Tabuk Governorate is located along the north-west coast of KSA, closing to the Jordanian-Saudi Arabia border and facing Egypt across the Red Sea. It has a land area of 146.072 km2, 900 meters above sea level, and an Urban Area population of 687,000. Due to the ongoing Red Sea Project in this region, Tabuk has recently received a specific attention as one of the most working areas in KSA in attracting numerous workers from different nations. Moreover, Tabuk ranks as one of the biggest agricultural regions in the Kingdom with abundance of stagnant water and groundwater, palm trees, and wild herbs, and its weather is characterized with mild climate in the summer and cold and rainy climate in the winters, which collectively make it a good environment for the breeding of leishmaniasis\u00b4s insect (sandflies)-vectors.Study participants, data sources/measurement: in this retrospective study, we used the surveillance database for the reported CL cases at the Vector-borne Diseases Control Unit, Local Health Directorate, Tabuk province, KSA. Data related to nationality, gender, age, and year-by-year and month-by-month distribution of the reported CL patients during the period from January 2006 to December 2021 were collected and analyzed. To meet the study objectives and the case definition, the included CL patients are defined as a person who was in Tabuk region during the said period and his/her CL was diagnosed by the standardized laboratory leishmaniasis-diagnosis as described by The WHO [ The WHO ,23, and Statistical analysis: data analysis were done using SPSS Statistics software package version 20.0 . The Chi-square (\u03c72) test and Student \u201ct\u201d test or Mann-Whitney test were used for the categorical data and continuous variables as appropriate. A P-value of <0.05 was considered statistically significant.Funding sources: this research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.A total of 1575 CL patients, who were resident in the Tabuk region, KSA, during the period of 2006 to 2021, and were diagnosed and registered as CL patients, were analyzed here. The included CL patients had no repeated CL cases in their registers during the said period. Notably, as showed in As represented in Cutaneous leishmaniasis (CL); the most common clinical form of leishmanial diseases worldwide ,24,25, iThe burden of leishmaniasis, particularly its CL form, continues to be one of the major health problems in many regions of KSA, and the endemicity of the disease was reported in the Eastern, Riyadh, Al-Hassa, Aseer, Al-Qaseem, Ha'il, and Jazan regions of the country -21. In tThe present study focused on whether there were distribution variables among the reported CL patients related to their nationality, age, gender, and the annual and month-by-month records. The total results showed no significant difference related to the patients' nationality. On the other hand, gender-and-age-dependent variations were significantly observed here whereby the majority of the reported CL patients were in those with age group of 15-45 years, the disease affected males more than females. Most importantly, a continuous annual and month-by-month record of these CL patients; as an evidence of CL endemicity in this region, as well as the annual and month-by-month record variations, were also detected. In agreement with the present findings, various earlier studies have been conducted in the different regions of KSA and reported that the CL disease affected males more than females, and the younger aged and adult individuals more than the other group of ages, but there was no significant variation in terms of number of CL cases between the Saudis and non-Saudi expatriates ,28. The Study strengths and limitations: the present survey included a number of important variables such as the annual and month-by-month distribution of the reported CL patients and variations in terms of their number and their residency, gender, and specific age group. Nevertheless, it would have been preferable if additional factors were available in the records of these CL patients and are included here; such as variables related to their general clinical data, occupational hazardous, anatomical distribution and morphological typing of the detected CL, molecular (PCR) typing of the underlying causative Leishmani species, and type and outcome of the applied treatment.Data of the present study were drawn from the registers of the Saudi and non-Saudi patients with CL; who were resident (2006-2021) in the Tabuk region, KSA, and revealed the endemicity of this protozoan parasitic disease in this region. Further screening implements and large-scale monitoring studies are warranted to assess the entire epidemiological features of CL disease, and to improve the public health measures to thorough control its endemicity in KSA.There are scarce evidence in the literature related to the epidemiological trends of cutaneous leishmaniasis (CL) in the Tabuk region, Saudi Arabia;The leishmaniasis\u00b4s control program in KSA remains challenging.The present findings provide an insight into the endemicity of CL in the Tabuk region of KSA;The present findings raise the essentiality for improving the controlling polices against Leishmania infections and their related diseases in KSA."} +{"text": "The paper provides a short history of the operations research (OR) in Slovenia. Some major events and achievements are mentioned and briefly discussed. The period starts in the year 1964, with the first symposium on OR in Slovenia. In the following decades, there were several important milestones: (1) the start of master\u2019s and Ph.D. studies in OR in 1974, (2) the establishment of SSI-SSOR in 1992 , and (3) the start of a series of symposia in operations research in 1993. All these activities resulted in an extensive list of publications, projects, and monographs and international connections, proving that OR is still a vibrant field, which facilitates knowledge transfer from pure research to business applications. Although some experts in Slovenia were familiar with the OR before, we may say that the first OR related formal event in Slovenia was the symposium entitled \"Mechanografics (Data processing) and OR\" with 30 papers, organized in Ljubljana in 1964, by the Slovenian Association of Economists and the Chamber of Commerce of Slovenia. The symposium consisted of two parts, the first part was dealing with the problems of data collection and processing, while the second with OR. In the first part, representatives of various companies and institutions reported on the increase in the volume of data, which requires a more rational way of data acquiring and processing. The presenters reported on experiences and problems in data collection and processing. Some of them also provided practical experiences and theoretical concepts. This duality emphasized the solid connection between theory and practice in the years that followed. In the second part, OR was presented as a new science for decision support in many areas. Several practical examples were given from the fields of industrial production, location, transport, agriculture, investments, and healthcare, in which linear programming and a two-phase production process supported by linear programming dominated as methods. The papers were printed in the proceedings . Symposia were held every year in Herceg Novi. Between 150 and 200 participants from all over Yugoslavia took part in them, some from abroad as well. Contributions to SYM-OP-IS symposia were published in proceedings, which were regularly issued as part of each symposium. Researchers and practitioners from Slovenia have regularly participated at these conferences.In 1974, at the Faculty of Economics of the University of Ljubljana started a master's degree in Operational Research, which some years later grew into a doctoral degree in Information and Management Sciences. This was probably one of the most important milestones in OR development in Slovenia. The students of this study were graduates of economics, mathematics, physics, mechanical engineering, electrical engineering, construction, law, sociology, and others who, after completing their studies, worked or they still work in the field of operational research at universities, institutes and research departments of companies. At the same time, the first book, Outline to operations research was founded and the colleagues from Croatia organized the 1st international OR conference (KOI\u201991). Many experts from the field of operational research in Slovenia took part at this conference. Following the example of our colleagues from Croatia, at the end of 1992, Slovenia's operational research experts founded their own association, the Slovenian Section for Operational Research (SSOR) under the umbrella of the Slovenian Society of Informatics (SSI), i.e., SSI-SSOR, which has nowadays 107 members. SSI-SSOR is a forum for scientists and practitioners from all areas of OR and allied fields, across the disciplines and programs in resource management, networks, tools, such as linear and nonlinear programming, discrete and combinatorial optimization, stochastic decision-making, multi-criteria optimization, strategic games, inventory theory, graph theory, dynamic optimization, systems management, theory controls and others, information, and education. At its first meeting the founders of SSI-SSOR clearly outlined the goal of the group which is to pursue, support, and facilitate research, development, application, and education in the operations research area where mathematics, economics, computer science, statistics, environmental economics, and system theory, as well as some other disciplines, come together. Pursuing the interdisciplinarity and applied OR science remains the core activity of SSI-SSOR. These include increase the visibility and influence of OR, closer collaboration with educational institutions, industry, government, and international entities, as summarized in Zadnik Stirn and Drobne .Since the foundation of SSI-SSOR, its members have been active in several fields , EURO, IFORS, and many other entities and individuals.In 1993, SSI-SSOR, with the support of the Ministry of Science and Technology of the Republic of Slovenia and the Faculty of Economics, University of Ljubljana, organized the 1st Symposium on OR, SOR'93, on national level. The Report of SSI-SSOR on SOR'93 is available at http://fgg-web.fgg.uni-lj.si/~/sdrobne/sor/ which can also be accessed from the symposia website https://sor.fov.um.si/publications/. Detailed symposia reports are available at https://www.drustvo-informatika.si/sekcije-drustva. In all 16 Proceedings which are indexed in Current Mathematical Publications, Mathematical Review, Zentralblatt fuer Mathematik/Mathematics Abstracts, MATH on STN Internationaland CompactMath, INSPEC and others, in total 1014 reviewed papers presented at SOR symposia are issued. Maximal number of papers were presented at SOR\u201921, i.e., 118, while minimal at SOR\u201995, only 21. The average of presented and in Proceedings published papers is 63,375 papers per symposium. There were all together 100 keynote papers presented by authors coming from 26 countries. The papers at the symposia were divided into sessions. At all symposia 157 sessions were performed, maximal 19 at SOR\u201921, average were 9.81 sessions per symposia. There presented and published papers were written by 1768 authors. Maximal number of authors are tracked at SOR\u201921 (240), minimal at SOR\u201995 (28), average number of authors is 110.5 per symposium. The authors come from all over the world, of course the most from Slovenia and geographically close countries . Maximal number of countries is exhibited at SOR\u201917 (25), minimal at SOR\u201993 , the average number of countries per symposium is 12.56.The symposia SOR are the premiere scientific event around OR in Slovenia that are providing an international forum for scientific exchange at the frontiers of OR, mathematics, statistics, economics, engineering, education, environment, computer science, and other fields. Since 1995, SDI-SOR, in agreement with HDOI, has organized a symposium every other year, which means that SDI-SOR and HDOI alternate annually in organizing OR international symposia, one year in Slovenia, the other year in Croatia. Thus, SDI-SOR organized: SOR\u201993, SOR'94, SOR'95, SOR'97, SOR'99, SOR'01, SOR'03, SOR'05, SOR'07, SOR'09, SOR'11, SOR' 13, SOR'15, SOR'17, SOR'19, SOR'21. The papers presented at these symposia were reviewed and published in Proceedings (2021), , 2017),,2017), , 2013),,2013), , 2009),,2009), , 2005),,2005), , 2001),,2001), , 1997),,1997), , 1993)..1993). PMost of the members of SSI-SSOR are attached to the universities or institutes. Thus, the members are mostly researchers in operations research and related fields. Some of them, together with their colleagues from abroad, mostly from Croatia, have compiled their work in the form of monographs. Since 1998, SSI-SSOR has published 5 monographs: Rupnik , Zadnik Islovar (http://islovar.org/islovar) which is recognized also as a reference dictionary for public users.As we can see from references, some monographs or their parts are in Slovene language. Here we would like to add that for professional Slovene language, i.e. Slovene terminology in operations research, the publications in Slovene are indispensable and of great importance. The members of SSI-SSOR take part also by editing and reviewing of the internet interactive IT terminology dictionary named In 1994, the SSI-SSOR joined the Austrian, Czech, Croatian, Hungarian and Slovak Societies for Operations Research in the initiative to start a high-quality international journal. Under the leadership of Austrian colleagues, the journal Central European Journal for Operations Research and Economic was initiated and it was published until the end of 1998. In the year 1999 the journal began to be published by Physica, and later by Springer Verlag under the current name Central European journal of Operations Research (CJOR). The journal was first indexed in Web of Science in 2007. Currently, it is ranked on the boarder of the first and second half of journals according to the impact factor in the Web of Science category Operations Research and Management Science , and is indexed at Portal of Croatian Scientific and Professional Journals. Currently, it is indexed as Q3 for Economics, Econometrics and Finance in Scopus. SSI-SSOR edited since 2012 six special issues: in 2012, 2014, 2016, 2018, 2020 and 2022. The special issues of BSR, i.e., SI of the BSR focused on recent advances in Operations Research and Management Science (OR/MS), with a particular emphasis on linking OR/MS with other areas of quantitative and qualitative methods in the context of a multidisciplinary framework to edit a special issue. https://uporabna-informatika.si/ui which is a Slovenian professional journal in informatics/OR. Its mission is to inform the professional public and users with up-to-date achievements in informatics/OR in Slovenia and worldwide. A special merit of the journal is its information of Slovenian research projects, and European documents which are the basis of trends in informatics/OR and inevitably influence our environment.Members of SSI-SSOR are also involved in publication of the SSI journal Uporabna informatika The diversity of applications of OR is reflected in the selection of papers published in this volume. The special issue call for papers has been synchronized with the organization of the 16th International Symposium on Operations Research in Slovenia, SOR\u201921. The conference was planned to take place at Bled, but had to be completed online due to limitations caused by the COVID-19 pandemic. However, the call for papers was not restricted to conference participants and all the manuscripts had been subject to the standard review procedure of the journal.Theoretical foundations of various models have always been an important part of OR, and often in the focus of temporary research. Linear programming with the simplex algorithm, is the first method that marked OR as a new academic discipline. Linear programming may still be seen as the leading methodology of OR. Several papers in this issue deal with various extensions of linear programming. Optimization problems are often subject to various kinds of inexactness or inaccuracy of input data. This leads to studies of varieties of classical problems where uncertainty is modelled with different means. Another usual generalization of the classical problems is the multi-objective programming, which is considered in several papers in this volume.The development of models and methods for solving the optimization problems to support the decision-making process is continuous. In this volume, several papers study advanced methods for optimization. Nagy and Varga investigate a new primal\u2013dual long-step interior point algorithm for linear optimization (E.-Nagy and Varga Because of its emphasis on practical applications, it is hard to decide which classification of contributions to OR is more justified. Sometimes, the application seems to be more important or, at least, more appealing, than theoretical considerations. In the years when we are facing a crisis of energy supply, it is of vital importance to predict, for example, electricity consumption, thus hopefully allowing higher quality of service and reliable delivery (\u010cegovnik et al. In this paper, we summarize the decades-long development of OR in Slovenia, which started in 1964 with the first OR symposium in Slovenia. An important milestone for the development of OR was the OR master\u2019s and Ph.D. programs at University of Ljubljana, which generated generations of OR specialists who are still important nodes in the OR community. Another important milestone was the founding of the Slovenian OR society in 1993 and the start of a series of operations research symposia SOR that will celebrate their 30th anniversary in 2023. These events, together with joining international OR organizations, are the cornerstones of international relevance of Slovenian OR. The future of Slovenian OR will strongly depend on the ability to motivate new generations to study Operations Research and to conduct research in OR or in other scientific domains using OR methodology.For this, the continuation of the SOR series is very important, as well as a strategic approach to attracting new talents in the field of OR."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "It is a good predictor of morbidity, mortality, and other health outcomes (3). Moreover, it has been effectively used in studies to document social inequalities in health (5). The coronavirus disease 2019 (COVID-19) outbreak measures have had a large impact on people\u2019s lives. In Japan, the government declared a state of emergency in seven prefectures on April 07, 2020. A nationwide emergency was imposed on 16 April 2020 and ended on May 25, 2020. This study aimed to compare the factors related to SRH before and during the spread of COVID-19. To reveal how SRH changed during COVID-19 outbreak in Japan, we examined SRH change before and during the COVID-19 outbreak by a secondary analysis of the data in the \u201cSurvey of Public Awareness for Food and Nutrition Education\u201d conducted by the Cabinet Office of Japan in 2018, 2019, and 2020.Self-rated health (SRH) status is correlated with objective health indicators We used individual data from the three independent \u201cSurveys of Attitudes toward Food and Nutrition Education,\u201d conducted by the Cabinet Office of Japan in 2018, 2019, and 2020 . The three surveys were performed in October 2018, November/December 2019, and December 2020. We used the data of S-2018-2019 and S-2020 as those before and during COVID-19 outbreak, respectively.For each nationwide survey, a two-stage stratified random sampling was used to select 3,000 people aged 20 or older. The survey sample was representative of the Japanese population and accounted for regional differences across the country, and the number of people surveyed in each municipality was proportionate to the municipality\u2019s population. Therefore, we assessed that the survey sample was appropriate to estimate SRH before and during COVID-19 outbreak. There were 1824, 1721, and 2395 participants for S-2018, S-2019, and S-2020, respectively. After removing missing data, 1823, 1709, and 2364 respondents provided adequate data for further analyses.We used the following questions, all of which were the same for the S-2018, S-2019, and S-2020. SRH was assessed based on the question: \u201cHow would you rate your general health status?.\u201d The responses were rated on a five-point Likert scale . Subjective economic status (SES) was assessed based on the question: \u201cHow would you rate your economic status?.\u201d The responses were rated on a five-point Likert scale . Healthy behaviors for prevention of lifestyle diseases were assessed based on the question: \u201cFor the prevention and improvement of lifestyle diseases, to what extent do you behave? For example, weight management, reduced salt, and so on.\u201d The responses were rated on a five-point Likert scale . Residential area was categorized into five city sizes: big city, medium-sized cities , medium-sized cities , small cities , and towns and villages.To analyze the differences in the distribution of the items among the S-2018, S-2019, and S-2020, the chi-square test was used. In addition, to analyze differences in SRH, SES, and healthy behaviors among S-2018, S-2019, and S-2020, nonparametric tests were conducted because the data were nonnormally distributed. Binomial logistic regression models were used to obtain odds ratios (OR) and their 95% confidence intervals for SES and healthy behaviors as predictors of SRH, after adjusting for age, gender, and residential areas.All data were analyzed using SPSS version 28.0 , with a significance level of 0.05.Ethics approval was not required for this secondary analysis of data publicly available through the University of Tokyo\u2019s Center for Social Research and Data Archives, Institute of Social Science.Nonparametric test results showed that the difference analysis of S-2018, S-2019, and S-2020 had statistical significance, with different internal differences . The disThis study examined the factors related to SRH before and during the spread of COVID-19. SRH became worse during COVID-19 outbreak (S-2020) with higher association with SES and healthy behavior compared with before the outbreak (S-2018 and S-2019). However, the main limitation of this study was that we used individual data from several cross-sectional surveys independently conducted before and during COVID-19 outbreak to measure the effects of the outbreak on SRH. Since the data used in the analysis were not longitudinal, but pooled data from each survey year, we could not reveal real causal relationship.(7). Japan currently shows one of the highest levels of average life expectancy in the world, as well as comparatively low levels of health inequality than other developed nations (9). However, warning bells are sounding about health inequality due to economic inequality in Japan recently (10). It is suggested that people with bad SES due to financial and job losses are more vulnerable to the COVID-19 outbreak (11). Holding constant COVID-19-related stress and background controls at both individual and contextual (country) levels, higher income is positively associated with better subjective health (12). Under unprecedented situations, such as the COVID-19 outbreak, healthy behaviors based on higher health literacy could be helpful in achieving good SRH even though SES condition is relatively poor. It is necessary to consider strategies to promote health education to improve public health outcomes.The association between health inequality and socioeconomic status is well documented across various context and population NoneThis work was supported by JSPS KAKENHI grant number JP22K02139.The data for this secondary analysis, \u201cSurveys of Attitudes toward Food and Nutrition Education,\u201d conducted by the Cabinet Office and the Ministry of Agriculture, Forestry and Fisheries of Japan in 2018, 2019, and 2020 were provided by the Social Science Japan Data Archive, Center for Social Research and Data Archives, Institute of Social Science, The University of Tokyo.HI: data analysis and manuscript writing; YI: project development, data management, data analysis, and manuscript writing/editing."} +{"text": "Int Braz J Urol is the 24th under my supervision and is very special. In July 23 the Web of Science released shows the 2022 impact factor and the Int Braz J Urol head the impact of 3.7 \u2013 the biggest in its history! A great reward for the hard work by the editorial team. With the new impact factor, the Int Braz J Urol today is one of the top 10 urologic journals (The September-October number of journals , an unimIn this number the Int Braz J Urol presents original contributions with a lot of interesting papers in different fields: Robotic Surgery, Overactive bladder, Endometriosis, Penile Cancer, Enuresis, Urolithiasis, BPH, Ureteroplasty and Varicocele. The papers came from many different countries such as Brazil, Italy, Russia, USA and China, and as usual the editor's comment highlights some of them. The editor in chief would like to highlight the following works:Dr. He and collegues from China, presented in page 535 a nice mDr. Diniz and collegues from the Urogenital Research Unit \u2013 Rio de Janeiro - Brazil, presented in page 564 a importDr. Tobias-Machado and collegues from Brazil performed in page 580 a nice sDr. Dahan and collegues form Brazil performed on page 590 an interDr. Xie and collegues form China performed on page 599 (Dr. Maida and collegues from Italy performed on page 608 (Dr. Guliev and collegues from Russia performed on page 619 an nice The Editor-in-chief expects everyone to enjoy reading."} +{"text": "Current trends in environmental psychology - volume II\u201d, is associated with the 3rd International Conference of Environmental Psychology (ICEP 2021), which was held in Siracusa, Italy, from the 4th to 9th October 2021 and it is the natural prosecution, and completion, of the volume I of the same Research Topic in a student sample from five countries , offering a psychometric contribution in line with recent suggestions by Tam and Milfont and how these changed in the post-COVID era. The authors presented a very informative case study: they conducted a choice experiment with individuals intercepted in the Argentera Valley, in the Western Italian Alps, highlighting a strong interest in biodiversity and cultural services, such as landscape aesthetic quality and psychophysical health. These findings could be useful to optimize the matching of supply and demand and to provide more robust information for promoting the participatory and shared decision-making process in forest planning and management. The importance of nature was investigated also with a theoretical contribution: Prins et al. conducted a systematic review and meta-ethnography of qualitative research on the value of play in nature-based compared to non-nature-based environments, and its implications for the developmental outcomes of young children (2\u20138 year). Their study showed that playing in nature-based environments supports young children's healthy global development, i.e. physical, social-emotional, motor, and cognitive. These results could be a further interesting insight to understand the dynamics and processes of humans-nature connectedness.Outdoor environments were also considered with different approaches: Haji and Hayati aimed to provide a comprehensive theoretical framework in the field of analyzing conflict behavior among rangeland exploiters in Iran. Specific environmental psychological theories, such as the Norm Activation Theory, the Value Beliefs Norms Theory, or the Theory of Planned Behavior, were found a suitable framework to explain conflict behavior in rangeland exploitation contexts. Likewise, using the Social Identity Model of Collective Action, Valizadeh et al. investigated Iranian farmers' intentions to participate in Aquifer Storage and Recovery, an innovative and alternative method for the sustainable management of water resources. The authors highlighted the need to consider the formation of social identity and the consideration of \u201cwe\u201d thinking systems as the best strategy for aquifer storage and recovery. The role of personality factors in shaping pro-environmental behaviors was also investigated by other studies: Haefner et al. investigated the mediating role of animal-related ethical values in the association between Big Five Personality traits, animal-related ethical values, and different types of meat consumption , providing useful information for susrtainable dietary change.Other studies have applied socio-environmental theories and models to a wide range of issues and topics: Suseno and Hastjarjo investigated the role of simulated natural environments in virtual reality and 2D video in reducing stress.Finally, digital environment studies could not miss the call in this RT: Researchers from Europe , Russia, China, Iran, and, Indonesia have contributed to the co-construction of a collective scientific endeavor that at this moment has collected about 25,000 views across 11 different papers.The richness and diversification of the published articles were the natural answers to complex questions like the ones presented in the call for papers of our Research Topic. A commitment to trans-disciplinarity was emerging and it could be seen as a form of cooperative research among the different parts of society, professionals, and academia (Pohl and Hadorn, OM: Conceptualization, Writing\u2014original draft, Writing\u2014review and editing. FF: Conceptualization, Supervision, Writing\u2014review and editing, Writing\u2014original draft. SM: Conceptualization, Supervision, Writing\u2014review and editing, Writing\u2014original draft. YP: Conceptualization, Writing\u2014review and editing, Writing\u2014original draft. MSa: Conceptualization, Supervision, Writing\u2014review and editing, Writing\u2014original draft. MSc: Conceptualization, Supervision, Writing\u2014review and editing, Writing\u2014original draft. GC: Conceptualization, Funding acquisition, Project administration, Supervision, Writing\u2014review and editing, Writing\u2014original draft."} +{"text": "Over 200 health journals call on the United Nations (UN), political leaders and health professionals to recognise that climate change and biodiversity loss are one indivisible crisis and must be tackled together to preserve health and avoid catastrophe. This overall environmental crisis is now so severe as to be a global health emergency.The world is currently responding to the climate crisis and the nature crisis as if they were separate challenges. This is a dangerous mistake. The 28th Conference of the Parties (COP) on climate change is about to be held in Dubai while the 16th COP on biodiversity is due to be held in Turkey in 2024. The research communities that provide the evidence for the two COPs are unfortunately largely separate, but they were brought together for a workshop in 2020 when they concluded that: \u2018Only by considering climate and biodiversity as parts of the same complex problem\u2026can solutions be developed that avoid maladaptation and maximize the beneficial outcomes\u2019.As the health world has recognised with the development of the concept of planetary health, the natural world is made up of one overall interdependent system. Damage to one subsystem can create feedback that damages another\u2014for example, drought, wildfires, floods and the other effects of rising global temperatures destroy plant life and lead to soil erosion, and so inhibit carbon storage, which means more global warming.Nature has a remarkable power to restore. For example, deforested land can revert to forest through natural regeneration, and marine phytoplankton, which act as natural carbon stores, turn over one billion tonnes of photosynthesising biomass every 8\u2009days.Restoring one subsystem can help another\u2014for example, replenishing soil could help remove greenhouse gases from the atmosphere on a vast scale.Human health is damaged directly by both the climate crisis, as the journals have described in previous editorials,Access to clean water is fundamental to human health, and yet pollution has damaged water quality, causing a rise in waterborne diseases.Changes in land use have forced tens of thousands of species into closer contact, increasing the exchange of pathogens and the emergence of new diseases and pandemics.Communities are healthier if they have access to high-quality green spaces that help filter air pollution, reduce air and ground temperatures, and provide opportunities for physical activity.Finally, the health impacts of climate change and biodiversity loss will be experienced unequally between and within countries, with the most vulnerable communities often bearing the highest burden.In December 2022 the biodiversity COP agreed on the effective conservation and management of at least 30% of the world\u2019s land, coastal areas and oceans by 2030.Yet many commitments made at COPs have not been met. This has allowed ecosystems to be pushed further to the brink, greatly increasing the risk of arriving at \u2018tipping points\u2019, abrupt breakdowns in the functioning of nature.This risk, combined with the severe impacts on health already occurring, means that the WHO should declare the indivisible climate and nature crisis as a global health emergency. The three preconditions for the WHO to declare a situation to be a public health emergency of international concernTackling this emergency requires the COP processes to be harmonised. As a first step, the respective conventions must push for better integration of national climate plans with biodiversity equivalents.Health professionals must be powerful advocates for both restoring biodiversity and tackling climate change for the good of health. Political leaders must recognise both the severe threats to health from the planetary crisis as well as the benefits that can flow to health from tackling the crisis.BMJ; Parveen Ali, Editor-in-Chief, International Nursing Review; Virginia Barbour, Editor-in-Chief, Medical Journal of Australia; Thomas Benfield, Editor-in-Chief, Danish Medical Journal; Kirsten Bibbins-Domingo, Editor-in-Chief, JAMA; Stephen Hancocks, Editor-in-Chief, British Dental Journal; Richard Horton, Editor-in-Chief, The Lancet; Laurie Laybourn-Langton, University of Exeter; Robert Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Peush Sahni, Editor-in-Chief, National Medical Journal of India; Wadeia Mohammad Sharief, Editor-in-Chief, Dubai Medical Journal; Paul Yonga, Editor-in-Chief, East African Medical Journal; Chris Zielinski, University of Winchester.Kamran Abbasi, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-nature-emergency-editorial-october-2023This Comment is being published simultaneously in multiple journals. For the full list of journals see:"} +{"text": "Mycobacterium tuberculosis as the causal agent of tuberculosis (TB) [On 24 March 1882, Robert Koch announced his discovery of In 2015, European countries endorsed target 3.3 of the Sustainable Development Goals (SDG) that aims at ending the TB epidemic by 2030 [Three years later, in September 2018, global leaders at the first United Nations (UN) General Assembly High-Level Meeting on the Fight Against TB agreed on ambitious targets to be achieved by 2022. These include increased access to TB prevention and treatment and increased funding for both TB services and TB research, to accelerate progress towards the 2030 goal .Eurosurveillance, Cristea et al. present their analysis of the progress achieved between 2018 and 2021 in the EU/EEA towards the 2030 targets for TB elimination [But since then, progress to end TB has stalled, even reversed, with geographically uneven performance across the World Health Organization (WHO) European Region. While most countries in Western Europe appear to be on track for TB elimination, Eastern European and Central Asian countries continue to experience a high burden of drug-resistant (DR) TB . TB inciWorryingly, 22% of the global cases of multidrug-resistant or rifampicin-resistant TB (MDR/RR-TB) and 42% There are multiple reasons why targets set in 2015 and then in 2018 have not been achieved. Consecutive crises such as natural disasters, conflicts, and the COVID-19 pandemic have severely impacted health systems, and jeopardised gains made, including the control of MDR/RR-TB and TB-HIV co-infection. Fragmented approaches and working in silos rather than collaborating and integrating efforts and resources, have slowed down success in Europe and globally.Just 3 years ago, the WHO European Region was on a good trajectory, with the fastest decline in TB incidence and mortality globally . By 2020The ongoing war in Ukraine has caused an additional setback, triggering an escalating humanitarian crisis in Europe with direct and indirect impacts on people\u2019s lives and health, as well as on Ukraine\u2019s health system. As a consequence, there is a risk of increased HIV and MDR/RR-TB infection rates in Ukraine and in Europe more widely, due to limited access to diagnosis and treatment and to population movement ,12. GuthThe economic impact of the war in Ukraine across Europe may affect governments\u2019 abilities to fund the commitment needed to scale up the TB response.The challenges are enormous, and they demand a determined response. To get back on track, we need revitalised and urgent political and financial commitment, and an agile and adapted response to TB. We also need to handle new challenges and threats proactively. For example, the article by Mart\u00ednez-Lirola et al. in this issue reminds us that TB must also be considered within the One Health approach; by using whole genome sequencing the authors uncovered hidden transmission routes for TB in Spain .In our favour, we have several new tools to support prevention and control of TB and DR-TB in Europe. For the first time, the duration of MDR/RR-TB treatment is 6 months \u2013 the same as for drug-susceptible TB, shortened from treatment of 18 months or longer; an important breakthrough in TB response. Further, these shorter, injection-free treatment regimens for MDR/RR-TB will substantially increase coverage and treatment success , while nTo truly end TB, we must interrupt transmission by finding people with active TB before they infect others and prevent those already infected from developing the disease. TB-preventive treatment (TPT) remains the main public health intervention and top priority for elimination, particularly for population groups at high risk of infection and progression from infection to disease, including people living with HIV and/or in contact with pulmonary TB patients . Providi\u201cending the spread of drug-susceptible and drug-resistant TB by achieving universal access to prevention, diagnosis and treatment in all Member States of the Region\u201d [In September 2022 there was a renewed political commitment for ending TB in Europe when countries endorsed the 2023\u20132030 TB Action Plan [ Region\u201d .Advancing science, finance and innovation, and their benefits, to urgently end the global tuberculosis epidemic, in particular by ensuring equitable access to prevention, testing, treatment and care [The year 2023 provides another opportunity to renew global TB commitments. On 22 September, during the UN General Assembly, world leaders will gather in New York to participate in a follow-up high-level meeting on TB. The theme of the meeting is For our part, the European Centre for Disease Prevention and Control and the WHO Regional Office for Europe, are committed to supporting all efforts and work towards ending TB in Europe by 2030 and beyond."} +{"text": "Subversive environmental impacts and limited amounts of conventional forms of energy necessitate the utilization of renewable energies (REs). Unfortunately, REs such as solar and wind energies are intermittent, so they should be stored in other forms to be used during their absence. One of the finest storage techniques for REs is based on hydrogen generation via an electrolyzer during abundance, then electricity generation by fuel cell (FC) during their absence. With reference to the advantages of the proton exchange membrane fuel cell (PEM-FC), this is preferred over other kinds of FCs. The output power of the PEM-FC is not constant, since it depends on hydrogen pressure, cell temperature, and electric load. Therefore, a maximum power point tracking (MPPT) system should be utilized with PEM-FC. The techniques previously utilized have some disadvantages, such as slowness of response and largeness of each oscillation, overshoot and undershoot, so this article addresses an innovative MPPT for PEM-FC using a consecutive controller made up of proportional-integral (PI) and proportional-derivative (PD) controllers whose gains are tuned via the golden jackal optimization algorithm (GJOA). Simulation results when applying the GJOA-PI-PD controller for MPPT of PEM-FC reveal its advantages over other approaches according to quickness of response, smallness of oscillations, and tininess of overshoot and undershoot. The overshoot resulting using the GJOA-PI-PD controller for MPPT of PEM-FC is smaller than that of perturb and observe, GJOA-PID, and GJOA-FOPID controllers by 98.26%, 86.30%, and 89.07%, respectively. Additionally, the fitness function resulting when using the GJOA-PI-PD controller for MPPT of PEM-FC is smaller than that of the aforementioned approaches by 93.95%, 87.17%, and 87.97%, respectively. Mechanical ES: This includes ES in the form of kinetic, potential, or compression energy. The most frequently utilized schemes for ES as mechanical energy are flywheels and hydroelectric pump storage . Other mMagnetic ES: In this scheme, ES is performed through supplying DC current via a coil and creating a magnetic field. In most circumstances, a superconducting magnetic coil is employed . The cooChemical ES: In this scheme, ES is performed via chemical or physical suction, intercalation, electrochemical procedures, or chemical conversion . PresentElectrical ES: In this scheme, ES in the form of electrical charge is performed, i.e., obtained via electricity; this process is generally accomplished via capacitors or supercapacitors .Biological ES: These schemes in general store energy which has been produced through breakdown of glucose via enzymes ; neverthThermal ES: In this scheme, ES is performed via storing heat in a latent, sensible or absorption manner. These schemes provide good opportunities for waste heat recovery and for domestic cooling/heating techniques .Electrochemical ES: The storage of electrical energy generated via RE sources in the form of electrochemical energy using rechargeable batteries is commonly implemented. Unfortunately, the life span of rechargeable batteries is short, so they need to be continuously replaced, which adds to their cost. Fuel cells (FCs) are promising means for extracting the stored energy via intermittent REs in the absence of a combustion process . HydrogeThe replacement of traditional sources of energy based on fossil fuels with renewable energies (REs) is inevitable for environmental reasons and due to the gradual depletion of fossil fuels. REs are environmentally friendly and their sources are not exhaustible. The wind blows alternatingly so the wind speed varies continuously and in some cases is less than cut-in speed, i.e., the required speed to generate electrical energy. Similarly, solar energy is not available during the night and cloudy weather. Thus, the disadvantage of REs is that their sources, such as solar power and wind, are not available all the time. Consequently, REs should be stored to continually assure the existence of electrical energy ,2. A divsta) of PEM-FC and DU is adjusted to make the output voltage (Vo) track the voltage at MPP (VMP). The difference between the various approaches to MPPT by PEM-FC is the determination method of VMP for adjustment of DU via the DC-DC boost converter.For each group of operation conditions for PEM-FC, i.e., hydrogen pressure, cell temperature, and electric load, there exists a unique point on the current\u2013power (I/P) plot representing maximum power. Accordingly, the maximum power point tracking (MPPT) procedure is required for extraction of the maximum power from the PEM-FC at various operation conditions. The MPPT system is a DC-DC boost converter with an adjustable duty switch cycle (DU). The DC-DC boost converter is fed via the stack terminal voltage (Vo is repeatedly varied via varying DU by a fixed step (\u0394DU), the resultant power and voltage variations are observed and, accordingly, DU is increased or decreased in the next variation until reaching MPP. The authors of [MP is determined wherever the derivative of power of the PEM-FC stack FC track the current at MPP. In the prementioned methods, the power of PEM-FC is calculated via multiplying the measured values of Vo and IFC while, in the variable step size (VSS) [FC is only utilized to decrease the cost and complexity.In this regard, perturb and observe (P&O) ,13 has bthors of have utithors of and the thors of methods,thors of ,18 have ze (VSS) ,20,21, tsta and IFC are entered into the trained artificial neural network (ANN) to produce the DU of the DC-DC boost converter. Adaptive ANN based on a fuzzy inference system has been applied in [In ,23,24,25plied in ,27,28.The authors of ,30,31 haIn addition to the aforementioned controllers, numerous others have been employed for MPPT of PEM-FC, i.e., the proportional-integral-derivative (PID) controller optimized by numerous algorithms, such as salp swarm approach (SSA) , the parRecently, metaheuristic optimization techniques have been applied for numerous purposes. Three kinds of these techniques are employed: evolutionary algorithms, physics-based, and swarm intelligence techniques. The first kind is driven by relying on biological evolution, e.g., DE and artificial bee colony. The second kind is driven by relying on physical laws, e.g., equilibrium algorithm and Archimedes optimizer. The last kind is driven by relying on the manners of animal groups, e.g., PSO, SSA, and GWO.In this regard, the GJOA is suggested for adjusting the gains of the PI-PD controller. The GJOA is a metaheuristic optimizer that replicates the golden jackal\u2019s manner during hunting . GJOA waThe innovative employment of the PI-PD controller for MPPT of PEM-FC.The innovative application of GJOA for adjustment of the gains of the PI-PD controller.Comparing the acquired results using the GJOA-PI-PD controller for MPPT of PEM-FC with those based on the P&O approach, GJOA-PID, and GJOA-FOPID controllers in order to confirm its supremacy.The GJOA-PI-PD controller performance is validated through variations in hydrogen pressure, cell temperature, and electric load.The contributions of this article are:The remainder of the article is organized as follows: FCs are overviewed in 2) from each molecule of water (H2O). The relationship between electrical power (3/h) is stated in (1) [3), e is the number of electrons implied in the reaction and equals 2 for water splitting, F is Faraday\u2019s number , 2. The energy conversion process inside is clean, since FC exhaust is water vapor.d in (1) :(1)\u03b7e=PeFCs are mainly categorized according to their electrolyte. This categorization establishes the type of electrochemical reactions which occur inside the FC, the type of catalysts needed, the temperature limit of the FC, the fuel needed, and other features. These FC characteristics impact their appropriate purposes. The kinds of FC include proton exchange membrane fuel cell (PEM-FC), solid oxide FC, phosphoric acid FC, alkaline FC, molten FC, and direct methanol . CompariThe PEM-FC stack model has been densely illustrated in the literature. For a stack composed of as below ,21:(2)Vs) to (6) ,21.(3)EMP), which is reliant on fc to VMP using the DC-DC boost converter. In this article, we suggest an innovative MPPT for PEM-FC using the PI-PD controller, whose gains are tuned by GJOA. We begin with an explanation of the DC-DC boost converter in the next section.By reference to swi) of PWM.o is dependent on input voltage (Vi) and DU as stated in (8) [Vd in (8) ,33,40,48i and Vo, the value of DU can be derived from (8) as below:For known values of Vsta and VMP represent Vi and Vo, respectively. Since Vsta and VMP change continuously, then DU needs to be adjusted continuously. The suggested control strategy for adjusting the DU of the DC-DC boost converter for MPPT of PEM-FC is illustrated in the next section.When the DC-DC boost converter is employed for MPPT of PEM-FC, VMP and Vo in order to make Vo track VMP continuously, and then The GJOA is a swarm intelligence optimizer which imitates the hunting manner of golden jackals in wildlife. Their hunting group consists of females and males. There are three stages in their hunting manner: 1\u2014seeking and approaching the prey; 2\u2014surrounding and confusing the prey, stopping its movement; 3\u2014swooping on the prey.Throughout the initialization step, a group of prey locations matrix (ing (11) .(11)PreE is the escaping energy of the prey and is computed using (11) .(12)E=Evia (13) .(13)E1=If and (15) :(14)L1itand (17) .(16)rl=As the prey is fatigued due to the chase, E is diminished and meanwhile, when and (19) :(18)L1itThe updated location of the prey in (20) .(20)LitAll details of the GJOA can be found in . The MATIn this subsection, we formulate the FiFu to be minimized by GJOA while tuning the gains of PI-PD controller, which in turn adjusts the DU of the DC-DC boost converter for MPPT of PEM-FC.sta track The main aim of MPPT of PEM-FC is to make Proven in . Minimizroven in :(21)FiFuThe FOPID controller differs from the PID controller in that the order of both integration and differentiation is a fraction instead of an integer. The transfer function of the FOPID controller is stated in (22) .(22)Cs=o and IFC are measured and Psta is calculated via multiplying their values, then DU is changed by fixed \u0394DU, which leads to a change in Vo and, accordingly, Psta. The resultant changes in Psta (\u0394Psta) and Vo (\u0394Vo) are monitored. If \u0394Psta and \u0394Vo have the same sign, then DU is decreased by \u0394DU, otherwise DU is increased by \u0394DU in the next iteration. This procedure is repeated until \u0394Psta equals zero, i.e., it reaches MPP. P&O is an iterative approach for MPPT of PEM-FC. P&O is commonly utilized for its simplicity. Firstly, VThe efficacy and forcefulness of MPPT of PEM-FC based on the GJOA-PI-PD controller are endorsed via comparing its results with those of other approaches. The impact of variations in The simulation results have been obtained via MATLAB-R2021 in Windows 11.swi = 10 kHz, high fswi is chosen to downsize the capacitors and inductors, which causes a cost decrease, L = 69 mH, and C = 1500 \u03bcF. These settings of L and C are carefully selected to assure low ripples in Vo at the indicated fswi. The limits within which the parameters of GJOA-PID, GJOA-FOPID, and GJOA-PI-PD controllers are maintained during minimization of FiFu using GJOA are listed in The GJOA is operated with these parameters: pop = 10 and max_ite = 5. The MPPT is performed on a commercial typical PEM-FC, namely the Ballard Mark V, whose parameters are listed in Normal operating conditions of The values of optimized parameters of GJOA-PID, GJOA-FOPID, and GJOA-PI-PD controllers are listed in sta of the Ballard Mark V PEM-FC when three MPPT schemes, plus the proposed scheme, are applied. Specifically, the P&O approach, GJOA-PID, and GJOA-FOPID controllers are compared with the proposed GJOA-PI-PD controller. High overshoot exists in the response of Psta when the P&O scheme is employed. There are oscillations and slowness in the response of Psta when GJOA-PID, and GJOA-FOPID controllers are employed. The resultant values of rise time (tr) and percentage overshoot (POS) for various MPPT schemes are listed in r with the proposed GJOA-PI-PD controller is 0.391 s, which is less than that of the GJOA-PID, and GJOA-FOPID controllers but more than that of P&O. The criteria in comparison are that the MPPT scheme, which has the quickest response, the least oscillations, and the lowest overshoot, is preferred over other schemes. When these criteria are applied to the results revealed in The previous comparison is based on visual analysis of the results. On the other hand, the comparison based on the numerical results of ITAE confirms the preference for the GJOA-PI-PD controller over other schemes, as summarized in sta during a change in sta increases to new value then decreases with decrease of sta tracks the new MPP for new conditions. The new conditions in this case study resulted in a variation of In this subsection, the GJOA-PI-PD controller for MPPT of the Ballard Mark V PEM-FC is validated when sta during variation of sta decreases to its new value then increases with increase in sta tracks new MPP for new conditions. The new conditions in this case study are caused by change in sta.This part presents a justification for the GJOA-PI-PD controller for MPPT of the Ballard Mark V PEM-FC when sta during change in sta decreases to its new value then increases with decrease of sta tracks new MPP for new conditions. The new conditions in this case study result in variation of In this subsection, the GJOA-PI-PD controller for MPPT of the Ballard Mark V PEM-FC is justified when The I/P plot of PEM-FC varies with the operating conditions, namely"} +{"text": "There are errors in the Funding section. The correct Funding statement is: This project was funded by the Deanship of Scientific Research (DSR) at King Abdulaziz University, Jeddah, under grant no. G: 1467-247-1440. The authors, therefore, acknowledge with thanks DSR for technical and financial support. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-023-40547-1, published online 23 September 2023Correction to: The Acknowledgments section in the original version of this Article was omitted. The Acknowledgments section now reads:\u201cThis research was based upon work supported by grants from the National Science Foundation under Grant Nos 1933394, 2121164; and supported/partially supported by the USC Center for Neuronal Longevity (#PG1033624); Unrestricted Grant to the Department of Ophthalmology from Research to Prevent Blindness, New York, NY; Dr. Allen and Charlotte Ginsburg Institute for Biomedical Therapeutics; Dennis and Michele Slivinski; The USC Roski Eye Institute; Dr. Ramani Nathan; and The Retina Research Foundation's Gertrude D. Pyron Award. No funding sponsors were involved in the study design, collection, analysis, interpretation of data, writing of the report, or decision to submit the article for publication. The following authors have a provisional patent (US Patent App. 17/685186) through the University of Southern California (USC): Mark S. Humayun, Gianluca Lazzi, Bodour Salhia, Ben Yi Tew, Javad Paknahad, Alejandra Gonzalez-Calle.\u201dThe original Article has been corrected."} +{"text": "CellDiscovery (2023) 9:51Correction to: 10.1038/s41421-023-00565-9 published online 26 May 2023In this article, the authors have found an error in the affiliations. The first affiliation is incorrect. The correct one is \u201cThe First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, China.\u201d We are sorry for the inconvenience."} +{"text": "There is a wide range of application for nanotechnology in agriculture, including fertilizers, aquaculture, irrigation, water filtration, animal feed, animal vaccines, food processing, and packaging. In recent decades, nanotechnology emerged as a prospective and promising approach for the advancement of Agri-sector such as pest/disease prevention, fertilizers, agrochemicals, biofertilizers, bio-stimulants, post-harvest storage, pheromones-, and nutrient-delivery, and genetic manipulation in plants for crop improvement by using nanomaterial as a carrier system. Exponential increase in global population has enhanced food demand, so to fulfil the demand markets already included nano-based product likewise nano-encapsulated nutrients/agrochemicals, antimicrobial and packaging of food. For the approval of nano-based product, applicants for a marketing approval must show that such novel items can be used safely without endangering the consumer and environment. Several nations throughout the world have been actively looking at whether their regulatory frameworks are suitable for handling nanotechnologies. As a result, many techniques to regulate nano-based products in agriculture, feed, and food have been used. Here, we have contextualized different regulatory measures of several countries for nano-based products in agriculture, from feed to food, including guidance and legislation for safety assessment worldwide. The developed crops with enhance phenotypes through conventional plant breeding methods will not be enough to meet the immediate availability of food and fodder globally. However, these methods were unable to introduce features that are not currently present in many plant species , though at 400\u00a0mg/kg affected the nutritional components of the cilantro. ZnO NPs also showed less toxicity than bilk and ionic counterparts are outstanding alternative to overcome negative impact of conventional fertilizers because they reduce nutrient loss from fertilizers and application rate of fertilizers . Researco fruits . Joint arecta L. . ZnO NPsterparts . CuO andterparts . Thus, nterparts .Different types of pesticides has been used indiscriminately worldwide, owing to the growing demand of agriculture-based food product. This indiscriminate use of different pesticides, has been associated with unprecedented environmental damage due to contamination of soil, water and food, leading to harmful effect on non-target pest species and humans . Thus, i2-NPs, 2\u00a0g/kg of stored grain) showed 100% mortality against four stored product insects; Rhizopertha dominica, Tribolium castaneum, Sitophilus oryzae, and Orizaephilus surinamenisis provide a solution with its three characteristic features; to increase solubility, slow/targeted release and protection against premature degradation . Nanopesamenisis . Temperaat 26\u00a0\u00b0C . Carboxynfestans . Nanopernfestans . Nanometmulation . Thus, nPhaseolus vulgaris as compared to free GA3 are used in various ways in agriculture to improve crop production . Variousfree GA3 .Nanosensers has many beneficial aspects in agriculture such as real time monitoring of environmental conditions and stress, crop growth and diseases, pest attack and nutrient efficiency Chen an. DevelopFor sustainable agriculture, plant genetic engineering is crucial for enhancing crop output, quality, and resilience to abiotic/biotic stressors . Plant gSeveral nanoparticle-mediated transgenic delivery techniques and the plant biotechnology industry\u2019s crowded field of existing methods. Together with a mix of the many newly created technologies, some other intriguing approaches, such the CRISPR technology, might be used in the processes of changing crops. Unfortunately, a number of significant problems still need to be fixed . The majNanotechnology has been increasingly used in the agricultural sector for various purposes, such as enhancing crop growth, improving soil quality, and developing more efficient and targeted pesticide delivery systems . However1. Regulatory Oversight: Each country has its own regulatory framework for using nanotechnology in agriculture. In the United States, Pesticides are governed by the Environmental Protection Agency (EPA), while agricultural biotechnology products are governed by the U.S. Department of Agriculture (USDA). Nanotechnology in foods and pesticides are governed by the European Chemicals Agency (ECHA) and the European Food Safety Authority (EFSA) in the European Union.2. Risk Assessment: Regulatory bodies require risk assessment before approval of any nanotechnology-based agri-product. This includes evaluating the toxicity of the nanomaterials used, the potential for environmental release, and the impact on human health.3. Labeling: Regulatory bodies require labeling of agri-products that contain nanomaterials. This helps consumers make informed decisions about the products they purchase and use.4. International Standards: International standards have been developed to assure the safety and quality of nanotechnology-based agri-products. The International Organization for Standardization (ISO) has developed several standards related to nanotechnology, including ISO/TS 80004-1, which provides terminology and definitions for nanomaterials.5. Research and Development: Regulatory bodies encourage research and development of nanotechnology-based agri-products to ensure that the products are safe for human health and the environment. Overall, the safe use of nanotechnology in agri-products requires a collaborative effort between researchers, manufacturers, regulatory bodies, and consumers. It is important to continue to monitor and assess the risks associated with nanotechnology-based agri-products to ensure their safety and effectiveness . Some exTo address these concerns, regulatory bodies around the world have developed guidelines and regulations to ensure the safe use of nanotechnology in agri-products. Here are some of the key regulations and guidelines related to nanotechnology-based agri-products.Different countries have established various regulations and guidelines to ensure the safe use and development of nanotechnology-based agri-products . Here arUnited States of America: Nanotechnology-based agricultural products in the United States are regulated by a number of governmental organisations, including the Food and Drug Administration (FDA), the Environmental Protection Agency (EPA), and the United States Department of Agriculture (USDA). There are nanomaterials that have been approved by the US-FDA for use in food applications. The following are some examples:1. Titanium dioxide: This is a common food additive used as a whitening and brightening agent in various food products, such as candy, chewing gum, and powdered sugar. Nanoscale forms of titanium dioxide have been approved for use in food products .2. Silica: Nano-sized silica is used in some food products as an anti-caking agent, such as in powdered foods like coffee creamer .3. Zinc oxide: Nanoscale zinc oxide has been approved for use as a food colorant and as a dietary supplement .4. Iron oxide: Nanoscale iron oxide has been approved for use as a food colorant .In general, these agencies have a common goal of ensuring the safety and efficacy of nanotechnology-based agricultural products, while also ensuring that they are in compliance with applicable regulations.Here are some key laws, safety measures, and regulations related to nanotechnology-based agri-products in the United States:1. The Toxic Substances Control Act (TSCA): This law gives the EPA the authority to regulate the production, importation, use, and disposal of chemical substances, including nanomaterials. Nanomaterials used in agri-products fall under the TSCA, and companies are required to provide the EPA with information on the potential health and environmental effects of these materials.2. The Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA): This law regulates the registration and use of pesticides in the United States. Pesticides that contain nanomaterials must be registered with the EPA, and companies must demonstrate that the products are safe for use.3. The Food, Drug, and Cosmetic Act (FD&C Act): The FDA regulates the use of nanotechnology in food and cosmetics. The FD&C Act requires that food and cosmetic products be safe for use and properly labelled. The FDA also requires that manufacturers of nanotechnology-based products provide information about the safety and efficacy of these products.4. The National Organic Program (NOP): The NOP is a USDA program that regulates the use of organic labelling on agricultural products. Products that are labelled as organic must meet certain standards, including restrictions on the use of synthetic substances. The NOP does not specifically address the use of nanomaterials in organic products, but companies that produce organic products are still required to comply with all applicable regulations.5. The Nanotechnology Research and Development Act (NRDA): This law directs federal agencies to coordinate research and development efforts related to nanotechnology. The goal is to ensure that the risks and benefits of nanotechnology are well understood and that appropriate regulations are in place.1. Conducting rigorous safety testing: Companies should conduct thorough safety testing to identify any potential risks associated with nanomaterials used in their products.2. Labelling: Companies should properly label their products to provide consumers with information about the ingredients used in their products. Labelling only the ingredients is a necessary but not sufficient requirement for meeting regulatory guidelines. Apart from listing the ingredients, other relevant information such as the dosage, potential exposure routes, and any associated health risks should also be provided to ensure consumer safety. For instance, in the case of nanotechnology-based agri-products, the U.S. Food and Drug Administration (FDA) recommends that companies provide additional information about the nature and properties of the nanomaterials used, such as their size, shape, and surface area, to enable risk assessment and management. Companies are also advised to evaluate the potential exposure pathways and take measures to minimize exposure to workers, consumers, and the environment .3. Environmental impact assessments: Companies should conduct assessments to determine the potential environmental impact of their products.4. Training: Companies should train employees on the proper handling and disposal of nanomaterials to reduce the risk of exposure.In addition to these laws and regulations, there are several safety measures that companies can take to ensure the safety of nanotechnology-based agri-products, including.Overall, the regulation of nanotechnology-based agri-products in the United States is an evolving field, and companies must stay up-to-date on the latest laws, safety measures, and regulations to ensure the safety and efficacy of their products.United Kingdom: In the UK, the regulation of nanotechnology-based agri-products falls under the responsibility of several governmental agencies, including the Food Standards Agency (FSA), the Department for Environment, Food and Rural Affairs (DEFRA) and the Health and Safety Executive (HSE).One of the primary regulations governing the safety of nanotechnology-based agri-products in the UK is the Nanotechnology Safety Guidance produced by the HSE in 2011. This guidance provides information on the safe handling and use of nanomaterials in various industrial settings, including agriculture.The FSA is responsible for ensuring the safety and quality of food products, including those derived from nanotechnology. In 2014, the FSA published a report on the safety of nanomaterials in food, which recommended that the use of nanotechnology in food products be subject to risk assessment and evaluation. Additionally, DEFRA has issued guidance on the use of nanomaterials in agriculture, including the safe handling and disposal of nanomaterials in agricultural settings. This guidance was updated in 2018 to reflect the latest scientific knowledge on the potential risks associated with nanomaterials. Overall, the regulation of nanotechnology-based agri-products in the UK is a rapidly evolving field, with new regulations and guidance being issued on a regular basis to reflect the latest scientific understanding of the potential risks and benefits of nanotechnology.Europe: Nanotechnology-based agri-products, such as pesticides, fertilizers, and animal feed additives, are subject to various regulations in Europe to ensure their safety for human health and the environment. Here are some of the key laws and regulations for nanotechnology-based agri-products in Europe, along with their reference and year (1. Regulation (EC) No 1107/2009 - This regulation establishes the rules for placing plant protection products on the market in the European Union (EU). It requires that all plant protection products be authorized before they can be sold or used. The regulation also sets out the data requirements for the authorization of plant protection products, including those that contain nanomaterials (Year: 2009)2. Regulation (EC) No 396/2005 - This regulation establishes maximum residue levels (MRLs) for pesticides in or on food and feed derived from plants and animals. It also applies to pesticides containing nanomaterials (Year: 2005)3. Regulation (EC) No 1935/2004 - The general safety standards for products and materials that come into contact with food are established by this regulation. It applies to all nanomaterials used in food contact materials, including those used in agri-products (Year: 2004)4. Regulation (EC) No 767/2009 - This regulation establishes the rules for the authorization and marketing of feed additives in the EU. It also requires that all feed additives be safe for animals and the environment. The regulation applies to all feed additives containing nanomaterials (Year: 2009)5. Regulation (EU) No 2019/1009 - This regulation establishes the rules for the making available on the market of CE-marked fertilizers. It also requires that all fertilizers be safe for human health and the environment. The regulation applies to all fertilizers containing nanomaterials (Year: 2019)and year .1. RegulIn addition to these regulations, there are also guidelines and recommendations from various European agencies and organizations, such as the European Chemicals Agency (ECHA) and European Food Safety Authority (EFSA), on the safety assessment of nanomaterials used in agri-products.Canada: In Canada, nanotechnology-based agri-products are regulated under several laws and regulations to ensure their safety for consumers and the environment. Some of the key regulations and their corresponding references and years are.1. Canadian Environmental Protection Act, 1999 (CEPA): This act is the primary federal legislation for regulating the environmental and human health impacts of nanotechnology-based products, including agri-products. The CEPA provides the framework for the assessment and management of nanomaterials under the New Substances Notification Regulations .2. Food and Drugs Act (FDA): This act is Canada\u2019s federal legislation for regulating food safety and consumer health. The FDA provides the legal framework for ensuring the safety, quality, and efficacy of food products, including those that use nanotechnology. The FDA also sets out labeling requirements for food products that use nanomaterials .3. Pest Control Products Act (PCPA): This act is Canada\u2019s primary legislation for regulating pest control products, including those that use nanotechnology. The PCPA sets out the requirements for registering and labeling pesticide products, as well as the safety and efficacy requirements for these products .4. Canada Agricultural Products Act (CAPA): This act regulates the marketing and inspection of agricultural products in Canada. Under this act, agri-products that use nanotechnology are subject to inspection and quality control standards to ensure their safety for consumers .Australia: The use of nanotechnology in agriculture is a rapidly growing field, and in Australia, the regulation of nanotechnology-based agri-products is overseen by several regulatory bodies. Here are some of the relevant laws, safety standards, and regulations for nanotechnology-based agri-products in Australia regulates the registration and use of agrochemical products, including those that incorporate nanotechnology. In 2014, the APVMA released a guidance document on the regulation of nanomaterials in pesticides and veterinary medicines .2. Food Standards Australia New Zealand (FSANZ) is responsible for regulating the safety and labelling of food products, including those that use nanotechnology. In 2015, FSANZ published a risk assessment of titanium dioxide nanoparticles in food .3. Work Health and Safety (WHS) laws in Australia require that employers take reasonable steps to ensure the safety of workers who may be exposed to nanomaterials in the workplace. The National Industrial Chemicals Notification and Assessment Scheme (NICNAS) also provides guidance on the safe handling and use of nanomaterials .4. The Therapeutic Goods Administration (TGA) regulates the safety and efficacy of therapeutic products, including those that use nanotechnology. In 2015, the TGA released a guidance document on the regulation of medicines that contain nanomaterials .ustralia .1. AustrChina: In China, the regulation of nanotechnology-based agricultural products falls under the jurisdiction of several government agencies, including the Ministry of Agriculture and Rural Affairs (MARA), the State Administration for Market Regulation (SAMR), the National Health Commission (NHC), and the Ministry of Ecology and Environment (MEE). The following are some key laws, safety standards, and regulations related to nanotechnology-based agricultural products in China, along with their references and years of enactment .1. Regulations on the Safety Assessment of Agricultural Genetically Modified Organisms (MARA Order No. 7)\u20142001. This regulation sets out the safety requirements and procedures for the approval of genetically modified agricultural products, including those that utilize nanotechnology.2. Safety Requirements for Food and Food Additives Containing Nanomaterials (NHC No. 13)\u20142011. This guideline establishes safety requirements and evaluation procedures for food and food additives that contain nanomaterials, including those used in agriculture.3. Technical Guidelines for Safety Assessment of Nano-Scale Agricultural Products (MARA No. 198)\u20142014. This guideline provides a framework for the safety assessment of nanotechnology-based agricultural products, including their production, processing, and use.4. Administrative Measures for Safety Evaluation of New Varieties of Agricultural Genetically Modified Organisms (SAMR Order No. 8)\u20142020. This regulation outlines the safety evaluation procedures for new varieties of genetically modified agricultural products, including those that utilize nanotechnology.5. Measures for the Administration of Environmental Safety Assessment of Agricultural Genetically Modified Organisms (MEE Order No. 12)\u20142021. This regulation sets out the procedures and requirements for the environmental safety assessment of genetically modified agricultural products, including those that use nanotechnology.India: In India, the regulation of nanotechnology-based agri-products falls under the purview of various agencies and laws, including the Department of Biotechnology (DBT), the Ministry of Environment, Forest and Climate Change (MoEFCC), the Food Safety and Standards Authority of India (FSSAI), and the Indian Council of Agricultural Research (ICAR).1. The Environment (Protection) Act, 1986: This law empowers the MoEFCC to regulate the production, import, export, and use of hazardous substances, including nanomaterials .2. The Hazardous Waste Rules, 2016: These rules require the registration and authorization of facilities that generate, store, and dispose of hazardous wastes, including nanomaterials .3. The Food Safety and Standards Act, 2006: This law establishes the Food Safety and Standards Authority of India (FSSAI), which regulates the safety and quality of food products in India. The FSSAI has issued guidelines for the use of nanotechnology in food products, including agri-products : These guidelines provide a framework for the safety assessment of foods derived from genetically engineered plants and microorganisms, including those produced using nanotechnology.7. MoEFCC Notification on Manufacture, Storage and Import of 8. FSSAI Regulations on Food Additives (2011): These regulations specify the conditions for the use of food additives, including those derived from nanotechnology, in food products.9. ICAR Guidelines on Nanotechnology Research in Agriculture (2010): These guidelines provide a framework for the safe and responsible use of nanotechnology in agricultural research and development.10. Indian Pharmacopoeia Commission (IPC) Guidelines on Nanoparticle Characterization (2019): These guidelines provide a framework for the characterization of nanoparticles, including those used in the production of agri-products.Here are some laws, safety measures, and regulations related to nanotechnology-based agri-products in India (dbtindia.gov.in).Overall, the regulation of nanotechnology-based agri-products in India is still evolving, and there is a need for more comprehensive and coordinated regulatory frameworks to ensure their safety and efficacy.Note: The above laws, safety measures, and regulations related to nanotechnology-based agri-products in India may also be subject to additional guidance and policies issued by the respective regulatory agencies. It is important to note that these regulations are constantly evolving and subject to change, and there may be additional guidelines and standards at the local or regional levels.Nanotechnology have paved a way to find out the new strategies to develop novel methods to bring scientific interventions that enabled us to raise a quality product in the field of agriculture and production of agri-products. Although, there are few ill effects of the technology which need to be mitigated to make it a successful approach. Further, researchers or scientists need to work on green synthesis approach to make it more reliable and ecofriendly which is the utmost need of the society. Green synthesis technique does not require any toxic solvent as a capping and reducing agent which eradicates the environmental pollution. Nano priming of seeds can also be the one helpful technique in order to maintain the pace of sustainable agriculture through the development of nano-primed plants that bear alterations at the molecular level and produces ultimate modifications in the phytochemicals and physiological changes in the plant without causing any harmful effect to the environment and plant itself. Moreover, it is simple, cost effective and requires less energy. Bottom down method should be focused. Apart from this, primary screening needs to be done to decide the usage of optimal dose or concentration of the chemicals or extracts used. Mode of delivery of nanoparticles should be specifically monitored or framed so that it will not carry any toxic substance with it. The effectiveness of respective regulatory systems for handling nanotechnologies has been actively investigated by a number of nations worldwide. Overall, the safety measure and regulations for nanotechnology based agri-product need to be updated time to time as the research in this field continues to come out with the new scientific interventions and its output which need to be screened by the regulatory bodies."} +{"text": "Protein Recognition and Associated Diseases\u201d addresses the recent advances in computational methodologies for the analysis and identification of important residues for binding, scoring, and ranking of structural models of protein-protein complexes, protein-protein interaction networks, and their applications in life sciences and human health.Protein-protein interactions are essential for many biological functions in all living organisms including cell signaling, molecular switching, transporters, receptors, and immunity. For the past few decades, tremendous advancements have been made in order to understand the recognition mechanism of protein-protein complex formation, reconstruct protein-protein interaction networks of an entire organism, and/or complete biochemical pathways. These efforts are mainly focussed on the identification of interacting proteins, prediction of binding site residues at their interface, evolutionary conservation of protein-protein complexes, prediction of protein-protein complex structures by docking, predicting the binding affinity of protein-protein complexes, and assessing the mutational effects on strength of binding and diseases . RecentlBrysbaert and Lensink analyzes the performance of several centrality measures for identifying major interacting residues involved in protein-protein binding using binding affinity data of interface mutations. Johansson-\u00c5khe et al. propose a machine learning-based method for scoring and ranking peptide-protein complexes. It encodes the structure of the complex as a graph with evolutionary and sequence features as nodes and physical pairwise interactions as edges. Su et al. integrate protein-protein interaction networks and gene expression profiles for detecting pancreatic adenocarcinoma candidate genes. Karan et al. report the development of four genomic information-based prediction methods, namely, 1) interolog, 2) domain, 3) gene ontology, and 4) phylogenetic for identifying the interaction between Oryza sativa and Magnaporthe grisea in a whole-genome scale.The opening article by In essence, this Research Topic covers the exciting developments in the area of protein-protein interactions both at fundamental and application levels. It will be a valuable resource for computational biologists, biochemists, biophysicists, bioinformaticians, and researchers working in the field of protein-protein interactions, and for those working on the biological role of protein-protein interaction networks and their relation to disease.We would like to thank all the authors for their outstanding contributions. The guest editors thank the Editorial Assistant Dr. Sara Gomez Cabellos, Commissioning Specialist, and Dr. Rahila Esposito, Commissioning Manager for their help and support in successfully completing the Research Topic."} +{"text": "Atopic dermatitis (AD) is a condition frequently encountered in medical practices across the country. Arming ourselves with appropriate and safe treatment modalities to provide relief for this chronic and relapsing inflammatory condition is of utmost importance to our patients and their families. Utilizing topical calcineurin inhibitors (TCIs) for the treatment of AD not responsive to high-potency corticosteroids, or low-potency corticosteroids and localized to the face, eyelids, and skin folds of patients >2 years, is reasonable to include in common practice. Despite the FDA\u2019s Black Box warning, to date no evidence has been published linking the TCIs to an increased incidence of malignancy in either children or adults that establishes causation. The Canadian Society of Allergy and Clinical Immunology (CSACI) therefore recognizes that the benefits of TCIs should be carefully weighed with the theoretical risks in advising patients, and acknowledges that long-term studies remain in progress. The safety and efficacy of topical tacrolimus and pimecrolimus should therefore be considered when treating children and adults with AD in Canadian allergy and immunology practices. Atopic dermatitis (AD), a chronic and relapsing inflammatory skin condition, is often the first manifestation of atopy in children ,2. Up toThe hallmarks of AD include pruritus and ill-defined erythema, with edema and/or vesicles if acute, and lichenification if chronic. It has a predilection for the flexural creases, or, in the case of children under 4 years of age, the cheeks, forehead and extensor surfaces of the limbs . EffectiLow to intermediate-potency topical corticosteroids have long been utilized as first-line therapy for AD not adequately managed by emollients alone. However, potent fluorinated corticosteroids are not indicated for use on the face, eyelids, genitalia, intertriginous areas, or in young infants. Topical calcineurin inhibitors (TCIs), such as tacrolimus and pimecrolimus, are alternatives to corticosteroids for AD on the face, eyelids and skin folds that is unresponsive to low potency topical steroids .TCIs exert their anti-inflammatory action by inhibiting calcineurin-dependant T-cell activation, thus inhibiting the activation of pro-inflammatory cytokines and mediators of the allergic inflammatory reaction . They haA recent review article by Frankel et al. concluded that tacrolimus 0.1% ointment has been demonstrated to be as effective as mid to high potency class III-V steroids in the treatment of AD . Both taDespite evidence demonstrating efficacy, resistance to using the TCIs in management of AD is prevalent among clinicians. This anxiety arises in part from the release of an FDA Black Box warning in 2005, citing concerns that chronic intermittent use of TCIs could lead to an increased incidence of Hodgkin\u2019s and non-Hodgkin\u2019s lymphoma, as well as melanoma and non-melanoma skin cancers . Followitheoretical risk of increased malignancy with the use of TCIs. This was based primarily on animal data, case reports and the mechanism of action of the drugs . Of notA repair . More reA repair . To dateA repair ,14,17. TA repair ,19. PrelA repair . A recenA repair . NotablyIn summary, topical calcineurin inhibitors are effective treatments for atopic dermatitis, and the benefits of their use in the appropriately selected patient population outweighs the theoretical risk of increased malignancy.1. Low- to intermediate-potency topical corticosteroids are first-line therapy for AD. However, intermediate-potency topical corticosteroids are not indicated for long-term use on the face, eyelids, genitalia, and intertriginous areas.2. Topical calcineurin inhibitors (TCIs) are indicated for AD in patients 2 years of age and older.3. There is no current published evidence showing that TCIs clearly predispose to malignancy.CSACI: Canadian Society of Allergy and Clinical Immunology; AD: atopic dermatitis; TCI: topical calcineurin inhibitor; FDA: Food and Drug Administration; AAAAI: American Academy of Allergy, Asthma and Immunology; ACAAI: American College of Allergy, Asthma and Immunology; CDA: Canadian Dermatology Association.This position statement did not receive financial support from any industry sources.Dr. Audrey O. Segal has served on an Advisory board for Sanofi. Dr. Anne K. Ellis has served on the speaker\u2019s bureau for Merck, Pfizer and Sanofi, an Advisory board for Paladin Labs and Sanofi, and has received research grants from Adiga Life Sciences/Circassia Ltd. Dr. Harold L. Kim has served on the speaker\u2019s bureau for Astellas, AstraZeneca, Merck, Novartis, Pfizer and Takeda, and on Advisory boards for CSL Behring, Merck and Novartis.This Position Statement was the product of an ad hoc committee of the Canadian Society of Allergy and Clinical Immunology. Each of the credited authors contributed substantially throughout the planning, drafting and revision stages of the document, and all authors read and approved the final manuscript."} +{"text": "Constantin von zur Muhlen was inadvertently excluded from the by-line of the published article. Dr. von zur Muhlen should be listed as the 15th author and is affiliated with: 1 Atherogenesis Research Group, Department of Cardiology, University of Freiburg, Freiburg, Germany. The author's contributions are: Analyzed the data, Contributed reagents/materials/analysis tools, wrote the manuscript."} +{"text": "In all countries of Europe, the population is ageing. This shift involves information needs on a variety of ageing-related themes. For better insight into similarities and differences among European countries, there is an increasing platform to support the added value of comparative analyses on ageing across the European Union. As many survey data on ageing exist, the most fruitful way forward is to share datasets and to harmonise concepts, indicators, and methods as much as possible. However, harmonisation may involve various difficulties. Two examples are given.In the context of the EU fifth Framework Programme, the Comparison of Longitudinal European Studies on Ageing project was among the first projects to attempt harmonisation of data on health and quality of life. Post-harmonisation was undertaken using population-based datasets in six countries. An example is given of the harmonisation process of the concept of ADL disability, which allowed the comparison of disability-free life expectancy across the six countries. A North-South gradient was found, showing shorter disability-free life expectancies in Spain, Italy and Israel as compared to Finland, Sweden and the Netherlands. This gradient was suggested to be caused by differences in educational level and in family culture.http://www.eposa.org) on the personal and societal consequences of osteoarthritis in older people comprises a more recent attempt at harmonisation, using population-based cohort studies in six countries. Here, post-harmonisation was less successful in that the main variable, osteoarthritis (OA), was defined in too different ways. The heterogeneity of OA definitions hampers comparing prevalence rates, and possibly, associations of OA with quality of life. Therefore, in a follow-up project, new data collection in the six cohorts was carried out, with pre-harmonisation of measurement instruments, including a standardised clinical assessment of OA. Preliminary findings show differences in OA prevalence, with higher rates in Southern Europe than in middle and northern Europe.The European Project on Osteoarthritis (In conclusion, post-harmonisation may be a cost-effective approach to use existing data, but may not always lead to comparable data. Even in case of pre-harmonisation, for any findings from multi-country studies, it should be considered to what extent differences observed are still linked to differences in data collection or are indicative of real cross-national differences."} +{"text": "A significant problem in the study of mechanisms of an organism's development is the elucidation of interrelated factors which are making an impact on the different levels of the organism, such as genes, biological molecules, cells, and cell systems. Numerous sources of heterogeneous data which exist for these subsystems are still not integrated sufficiently enough to give researchers a straightforward opportunity to analyze them together in the same frame of study. Systematic application of data integration methods is also hampered by a multitude of such factors as the orthogonal nature of the integrated data and naming problems.Here we report on a new version of BiologicalNetworks, a research environment for the integral visualization and analysis of heterogeneous biological data. BiologicalNetworks can be queried for properties of thousands of different types of biological entities and their relations . The system includes the build-pathways infrastructure for molecular interactions/relations and module discovery in high-throughput experiments. Also implemented in BiologicalNetworks are the Integrated Genome Viewer and Comparative Genomics Browser applications, which allow for the search and analysis of gene regulatory regions and their conservation in multiple species in conjunction with molecular pathways/networks, experimental data and functional annotations.http://www.biologicalnetworks.org.The new release of BiologicalNetworks together with its back-end database introduces extensive functionality for a more efficient integrated multi-level analysis of microarray, sequence, regulatory, and other data. BiologicalNetworks is freely available at As substantial amounts of data concerning expression, interactions/pathways, sequences, and other types of information for a variety of tissues, developmental stages, stimuli and organisms are generated, it becomes difficult for researchers with no background in bioinformatics and statistics to extract the information they seek. Successful data integration is hampered by the orthogonal nature of the integrated data and by the multitude of controversies and name/ID conflicts in public databases. Examples of conflicts that cannot be automatically resolved include the situations when genes with the same name point to different chromosome locations or a gene/protein in different modification states has different names; for example, p53, p53361-393), p53(modified:Thr:212), or pCMX-mutant-p53. Among the name/ID conflicts that can be resolved is, for example, the conflict between different genes/proteins having the same synonym or the conflict between two databases naming the same gene differently - these and similar name/ID conflicts can be automatically resolved if there are other databases that recognize the conflicting names. To analyze and visually integrate publicly available data on the systems level, several web-based tools have been developed: Genomatix -393, p53, CytoscaIn this work, the application BiologicalNetworks 2.0 for integration of functional genomics data with biological networks is presented. In comparison with other tools . IntegromeDB data is stored in a Postgres database under the MetaGraph schema and is updated monthly, being synchronized with the latest changes in most current databases. Detailed information on the statistics, integrated databases catalog, and organism list can be found at http://www.integromedb.org.The backend database of BiologicalNetworks, called IntegromeDB , is a sehttp://www.w3.org/RDF/ and the Web Ontology Language (OWL) http://www.w3.org/TR/owl-ref/. IntegromeDB also enables researchers to integrate their own data into the database as described in the section 'Integration of User's Data.'The procedure of data integration and mapping to the internal database is fully automated and is based on the Semantic Web technologies, such as the Resource Description Framework (RDF) http://www.bioontology.org. The mapping among the OBO ontologies, which is provided by OBO, allows for the automatic integration of 98 ontologies in BioNets Ontology. The basic.owl file with Basic Ontology and mappings from it to other ontologies can be downloaded at http://www.integromedb.org/bionetsonto.php.The IntegromeDB schema is based on BioNets Ontology, the core of which is Basic Ontology that was manually developed by the authors. Basic Ontology describes classes from different domains, such as, protein, gene, pathway, interaction, disease, cell, tissue, drug, chromosome, COG functional group, gene set . Basic Ontology is manually mapped onto 25 OBO ontologies, including Sequence Ontology, GeneOntology, Human Disease, CheBI, BRENDA Tissues, that were selected from the best curated and regularly updated ontologies provided by the OBO consortium Due to the generic schema of the database and ontology-driven mapping, during integration, new objects and their properties are automatically added in the database. For example, if the database stored information about the interaction between the two objects, proteins \u00d7 and Y, new information about this interaction will be integrated as a new property(s), e.g., a property 'p-value from the experiment A', and the 'experiment A' will be added in the database as a new object. If a clear evidence of, or reference to, a class from the BioNets ontology is absent (missed), an automatic procedure to statistically evaluate the content of the integrated table and assign a term from the ontology is applied. The procedure takes each word and word combination in the table, search for them in the BioNets ontology, calculates the statistical significance of the occurrence, and assigns the most significant term to the table.http://www.integromedb.org. If the user searches the http://integromedb.org page for an object for which inconsistencies were calculated and found, they can be seen on the result search page by clicking the 'red button'.Upon the integration of a new data source, the database automatically identifies conflicts in names, name synonyms and IDs of all objects in the database among various databases and identification systems. All names/IDs are weighted and sorted by the number of integrated data sources, supporting the name/ID. When the user searches the database, names/IDs appear in the search result in the descending order by weight; the potential conflicts will appear in the bottom of the list. Thus, if one database, for example, names the specific gene as \u00d7 and another database, as Y, both names, \u00d7 and Y, will be equally searchable; while in the search result, the name that is the most common among all integrated databases will appear first. However, no conflict data are removed or become invisible to the user, since the data sources are not weighted or judged. Inconsistencies among the names/IDs of all objects in the database that were found for human, mouse and rat are provided at http://www.integromedb.org; however, in comparison with the application BiologicalNetworks, described in this work, the web site provides only general quick search capabilities and no extensive data analysis, dynamic integration, and visualization capabilities.The IntegromeDB data can be searched at The typical user of BiologicalNetworks starts with loading the file or searching the integrated database for a list of genes , networks (.sif-file), curated pathways (.SBML), microarrays, proteomics data (tab-delimited file), or sequence data . BiologicalNetworks provides an Integrative View of the found data Figure , openingMost current web-based tools are focused on retrieving expression and/or co-expression patterns for individual genes from particular microarray experiment. Multi-experiment/multi-gene co-expression analysis is a labor-intensive and computationally challenging task, involving collecting suitable datasets, data downloads, preprocessing, normalization, and gene annotation management, the integration of different datasets, merging cross-platform data, and handling ambiguous mappings between genes and probe sets. Microarray databases Gene Expression Omnibus (GEO) and Arrae.g. a set of transcription factors regulating a set of co-expressed genes), since several studies exploit the fact that co-expressed and/or functionally related genes may be transcriptionally coordinated [The user can upload the microarray data files and analyze them in conjunction with the integrated compendium of publicly available microarray data using the Microarray Analysis window. When searching in the Microarray Analysis window, the user can select the 'Default', 'Co-expression pairs' or 'Co-expression Triples' search modes. In the first case the search will return all available microarray experiments in which genes from the input gene list are over- or under-expressed. In the second case, it will return experiments and gene pairs where genes are co-expressed; and in the third case, it will return gene triples and experiments where these gene triples are co-expressed. This last option is especially important for discovering gene regulatory modules . Our gene list functional enrichment analysis currently uses as many as 8 annotation categories including 3 categories of GO terms, curated (e.g. KEGG) pathways, diseases, cell types, tissue gene expression, phenotypes and human anatomy in our internal database, we visualize them as tables ('Themes' Tables) for the sake of usability and easier navigation Figure . Since Bhttp://www.integromedb.org/bionetsonto.php nodes can be introduced and modified with minimal impact on the rest of the system. This is implemented through 'ontology mapping' [Gene, 'SO:12345') a new class that maintains mapping to source ontology is generated (i.e. class 'mappingSO:012345') which is connected to a Gene in our BioNets ontology through 'same_as' relation. This is done to not modify BioNets ontology classes every time new ontology is ingested, thus while 'unifying' different biological data types to keep specificity of every member schema of our integrated database. Examples of operations on RI-trees that will apply on all substructures , called SUB_X, are represented below:Sequence data together with annotation data, including binding sites, promoters, and other regulatory regions, that have been integrated in the BiologicalNetworks database represent the collection of interval trees; a single interval tree is created per chromosome instead of per annotated DNA sequence regions. Nodes of the interval (RI)-trees, or sequence intervals. are connected to the BioNets Ontology s Figure through mapping' : for eveifOverlap function: SUB-X * SUB_X - > {0, 1}, returns true if the two interval substructures overlap.Next function: SUB_X ->SUB_X is applicable on data types for which there is a strict ordering on the domain; it returns the sub-structure encountered next in the ordering input substructure. The semantics of 'next' depends upon the data types .Intersect function: SUB_X * SUB_X ->SUB_X, returns the intersection of two SUB-X. This operation is valid for convex data types such as sequences and rectangles.Navigation of segment elements and different annotations (properties) integrated from many data sources for one gene or gene upstream region. Genomic Sequences are integrated with the meta-graph schema of Biological-Networks database through an ElementId-ObjectID connection table, where elements are sequence elements, for example, a core promoter, TATA box, or binding site, that are attributed to a particular gene by means of known localization in the gene, according to the GenBank global position. Internal enumerations in the integrated databases-TRANSFAC, for example, provides localization of regulatory regions in respect to the transcription start-are recalculated accordingly. The connection table assigns sequence elements to meta-graph objects, so that sequence elements, represented as a RI-tree structure, become graph objects within the meta-graph database. All heterogeneous data, for example gene properties in the Property Panel Figure and Anno) Figure . Figure In the Sequence Analysis Browser, the user can upload large (GBs) volumes of sequence data and analyze them together with the integrated data on gene sequences, annotations, orthologs and cross-references to the major biological databases displayed in the Sequence Annotation Browser Figure . The broBiologicalNetworks provides the ability to investigate transcriptional cascades by integrating and visualizing transcription factor gene regulation networks, relevant transcription factor binding sites and target genes with multiple sequence annotations, thus facilitating validation experiments (e.g. primer design applications) Figure . A fundaThe Comparative Genomics Browser can be explored together with other modules/windows of BiologicalNetworks. For example, Figure The BiologicalNetworks interface contains multiple search and build pathways/networks capabilities Figure , allowinThe seven querying options listed below allow for the specification and search for any logical combination of entities, processes/relations and their properties. The first four options are available from Quick Search Box at the top right corner of the program Figure and alloSimple Gene/Protein search (default search). For example, the search for the three genes/proteins 'p53, egfr, esr1' returns these three genes in all specified species and gene properties. The list of genes/proteins can be uploaded from the file.1. Keyword search. For example, the keyword search 'p53, egfr, esr1' returns all database objects, including experiments, publications, pathways, and all properties of all objects that contain either p53, egfr, or esr1.2. Wild card search. For example, the search 'neurodeg*' returns all objects related to neurodegenerative, neurodegeneration, and all words in the databases beginning with 'neurodeg'.3. Multi-word search. For example, the search 'obesity diabetes' returns the results for 'obesity AND diabetes' and 'obesity OR diabetes'.4. Build Pathway Wizard: Path Queries. Build Pathway Wizard contains dozens types of pathway and network searches (in the opened networks/pathways or integrated database) in protein-protein interactions, transcription factor-DNA networks, relational networks and curated pathways (e.g. KEGG) Figure . The thr) Figure allow usa) algorithm type for pathway building;b) select the directionality of relations;c) types of objects and property values to be included in the pathway; andd) relations to be included in the new pathway.Specialized search or GEO experiments (Series and Datasets). The system recognizes most of the currently available gene/protein IDs and synonyms for thousands of organisms integrated from over 100 data sources. To perform a search in the microarray, pathways/networks, sequence annotations and PubMed repositories, the user can specify any combination of keywords, including authors names, tissue types, diseases, gene/protein names. The Search Box contains different configuration options and filters and enables limiting searches on specific species, opened network/pathway and sequences. The organism drop-down menus in each search window include 21 model organisms, which are mostly represented in the database, and the following options to narrow down the search and subsequent data analysis: All Organisms, Eukaryotes, Prokaryotes and Viruses. Eukaryotes are subdivided into Plants, Fungi, Protists, Archea, and Metazoa/Animals, which in turn are subdivided into Vertebrate, Invertebrate and Mammals.Search Boxes accept lists of gene names (that can be loaded from files), accession numbers from public databases , PSI-MI, Tab-delimited network file, SBML and BiGG model format;\u2022 microarray data: tab-delimited file format, Illumina tab-delimited and Affymetrix file formats;\u2022 sequences: GenBank and FASTA formats.The results of analysis and visualization in BiologicalNetworks can be saved at any moment as the BioNets XML Project file and then opened at other computers; the user's settings, data files, results of search, built networks, clustering, colorings and all other visualizations will appear exactly how they were at the moment of the file saving. The project can be also saved/exported to the SIF and SVG formats.http://integromedb.org.BiologicalNetworks allows the user to work with his/her own data, and to integrate them into the system database (IntegromeDB). The integration procedure is different from the procedure of loading/opening data files described above in that the loaded data are available to the user and also to whoever obtains the BioNets XML Project file that includes the user's data. The integration procedure allows the data to be made public; they become integrated in IntegromeDB as any other database and become searchable in the BiologicalNetworks application and at the web-page http://integromedb.org under \"User's Data Integration\" menu problems are minor problems that can be quickly fixed by the developers, if the Bug and Problem Report Tools are used.To address the critical need for user support, we developed Bug and Problem Report Tools. This tool allows the user to report problems or bugs, while working in BiologicalNetworks. During the installation of BiologicalNetworks and the initial run of the program, the user is asked for the agreement for permission to send from his computer any future bug reports. If the user agrees, Bug Reports will be automatically generated and sent to our support server; it will include the environment settings and the last user's steps before the program gave an error. To report the problem, the user needs to use Problem Report Tool that is located in the 'Tools' menu of the program. The Streptococcus pneumoniae -human/mouse/rat interactions [Giardia lamblia [Thermatoga maritime [The BiologicalNetworks analytical and querying functionality were applied to and tested in a number of different biological systems/projects, both eukaryotic and prokaryotic: host-pathogen interactions, specifically, the influenza and ractions , 'meta-gractions , yeast mractions , whole-gractions , parasit lamblia , and micmaritime . All desRattus norvegicus [In this section, we demonstrate a case study of the search for potential therapeutic targets for hypertension. Specifically, it is shown how, using BiologicalNetworks and starting with a single microarray experiment in the model organism rvegicus , one canFirst, among about 1000 genes significantly perturbed in hypertension in the microarray experiment Figure , we foun-3 and examined visually for consistency. In the result, we obtained 103 TFs that might potentially regulate transcription of 18 hypertension-specific genes . To construct the final network, we searched for all reported interactions among identified 103 TFs and 20 genes which can be replicated by simply running the respective project.http://www.biologicalnetworks.org.BiologicalNetworks, along with the user Manual and Video tutorials and Quick Start Guide, is available at Project name: BiologicalNetworksProject home page: http://www.biologicalnetworks.orgOperating systems: Windows 2000/XP/Vista/7, Linux/Ubuntu/Redhat, MacOSXProgramming language: JavaLicense: Free for academic purposesOther requirements: 2GB RAMMB, AG and MS contributed to system concept. SK, YD, MS and MB implemented the system and performed major programming work. MB and JP coordinated this work, contributed to data analysis and wrote the manuscript. All authors read and approved the final manuscript.Methods. Detailed description of the methods and data types used in the BiologicalNetworks system.Click here for file"} +{"text": "Raymond Hutubessy is a senior health economist affiliated to the Immunization, Vaccines and Biologicals (IVB) Department of the World Health Organization (WHO), and is the executive secretary of the WHO Immunization and Vaccines related Implementation Research Advisory Committee (IVIR-AC). His main research interests focus on economic and financial analyses of vaccine introduction decisions in low- and middle-income countries (LMICs). In this Q&A, he will discuss the importance of this work in relation to the global context of vaccine introduction decisions Figure .Vaccines for prevention of communicable diseases have been shown to be extremely effective in terms of health outcomes. Therefore, conducting economic analyses to get the most value for money from vaccine introduction decisions is of high importance; evidence and information resulting from these analyses are not the only input to the decision-making process for vaccine introduction decisions, but they are important ones.The relationship between health and economic growth is one of the cornerstones of development economics: health status is determinant of productivity that can be shown to influence economic growth. Specifically, vaccines have a broader value in terms of their indirect effects and other externalities . Therefore, in addition to the traditional economic appraisals for vaccine introduction decisions it is useful to policy makers and other stakeholders involved with vaccine introduction decisions to demonstrate the broader added value of vaccines and investments in health in general.Economic appraisals address different key issues with regard to decisions on vaccine introduction. These appraisals range from priority-setting issues across vaccines and other competing health interventions, to affordability and budget impact analysis, and costing and financing issues with regard to the introduction decisions of immunization programs. For these different policy questions, different analytical tools are available, such as cost-effectiveness analyses, costing studies, budget impact and optimization analysis.First, because many economic evaluations are based on analytical decision tools such as mathematical infectious disease models, costing tools, decision trees models and so on, transparency is needed on the choice of the modeling methodologies, parameters and country data used and assumptions made by the analyst. Standardization of methods of cost-effectiveness is therefore needed and analysts in the field should adhere to these guides. This allows users to make comparisons of different study results by different groups. The WHO, in addition to other organizations, has developed several guidelines on economic evaluations in health, and vaccines and immunization programs in particular.Second, to be relevant, local decision makers have country-specific policy questions and therefore need contextualized study results driven by specific country data and information with regard to demographics, epidemiological and economic data, and local needs. However, this does not mean that for each country , economists and analysts need to start from scratch - they can build on work from other groups who often put their models, including the computer program codes, in the public domain.In light of this, efforts should be put into the collection of local data and building local technical capacity in LMICs so that they are able to perform their own analysis and interpret their own results with the aim of increasing local ownership of the evidence generated. The WHO, along with partners i.e. Pan American Health Organization (PAHO), Agence de M\u00e9dicine de Pr\u00e9ventive (AMP), Program for Appropriate Technology in Health (PATH), Sabin Vaccine Institute and USA Centers for Disease Control and Prevention, recently started the ProVac International Working Group to promote the use of economic analysis for vaccine introduction decisions in LMICs.In principle, the methods applied and tools used are similar in higher income versus resource-limited settings. However, because country demographics, disease burden, epidemiological and socioeconomic background, and health systems and infrastructure differ, the methods of measurement and valuation and the interpretation hence key drivers of results of economic evaluations will also differ.For example, the price at which human papillomavirus (HPV) vaccination is considered to be cost-effective is heavily dependent on HPV prevalence and the existing local cervical cancer services and the ceiling cost-effectiveness or cost-effectiveness threshold . In high income countries, access to health care services is better and delivery systems of vaccines to reach adolescent girls are more advanced than in LMICs. As a result, the affordability question in such resource-rich scenarios focused around the relatively high public market prices of HPV vaccines, which may go up to 150 USD per dose.per se, it is the securing of the delivery costs to get the vaccine from the port of entry to those girls in need that has become a main barrier in many of these countries. This barrier is in addition to other capacity issues, such as the existing delivery infrastructure being already over-stretched with traditional Expanded Programme on Immunization vaccines and other competing new vaccines, such as rotavirus and pneumococcal vaccines.By contrast, in countries eligible for the Global Alliance for Vaccines and Immunization's (GAVI Alliance) support, the manufacturers of one of the vaccines have offered an indicative price of 5 USD per dose. This is a 64% reduction on the lowest public prices. As a result, rather than the vaccine price In my opinion, economic analyses have helped to increased the level of health that health care spending can buy, and hence have aided promotion of the use of effective and cost-effective health interventions. For example, by identifying barriers and uptake issues of vaccine introduction decisions, economic analyses have contributed to bridge the link between the evidence on theoretical vaccine efficacy and real-life effectiveness. Another example is that economic analyses do not just appraise vertical disease programs but more often also have an integrated and combined disease program perspective. This has the potential to account for synergistic effects of disease programs and therefore will improve the overall public health impact.WHO and other partners have vaccine specific and generic websites on health economic information and initiativies. In addition, WHO published several key documents on vaccine economics.WHO Initiative for Vaccine Research (IVR) [http://www.who.int/vaccine_research/implementation/health_economics/en/index.html]CHOosing Interventions that are Cost Effective (WHO-CHOICE) [http://www.who.int/choice/en/]WHO guide for standardization of economic evaluations of immunization programmes [http://whqlibdoc.who.int/hq/2008/WHO_IVB_08.14_eng.pdf]Pan American Health Organization (PAHO ProVac) [http://new.paho.org/provac/]International Health Economics Association [https://www.healtheconomics.org/]Economic analyses to support decisions about HPV vaccination in low- and middle-income countries: a consensus report and guide for analysts. BMC Med 2013, 11:23.Jit M, Levin C, Brison M, Levin A, Resch S, Berkhof J, Kim J, Hutubessy R: A case study using the United Republic of Tanzania: costing nationwide HPV vaccine delivery using the WHO Cervical Cancer Prevention and Control Costing Tool. BMC Med 2012, 10:136.Hutubessy R, Levin A, Wang S, Morgan W, Ally M, John T, Broutet N: Systematic review of studies evaluating the broader economic impact of vaccination in low and middle income countries. BMC Public Health 2012, 12:878.Deogaonkar R, Hutubessy R, van der Putten I, Evers S, Jit M: Comparative review of three cost-effectiveness models for rotavirus vaccines in national immunization programs; a generic approach applied to various regions in the world. BMC Med 2011, 9:84.Postma MJ, Jit M, Rozenbaum MH, Standaert B, Tu HA, Hutubessy RC: Results from evaluations of models and cost-effectiveness tools to support introduction decisions for new vaccines need critical appraisal. BMC Med 2011, 9:55.Hutubessy R, Henao AM, Namgyal P, Moorthy V, Hombach J: Human papillomavirus vaccine introduction in low-income and middle-income countries: guidance on the use of cost-effectiveness models. BMC Med 2011, 9:54.Jit M, Demarteau N, Elbasha E, Ginsberg G, Kim J, Praditsitthikorn N, Sinanovic E, Hutubessy R: Cost effectiveness of pediatric pneumococcal conjugate vaccines: a comparative assessment of decision-making tools. BMC Med 2011, 9:53.Chaiyakunapruk N, Somkrua R, Hutubessy R, Henao AM, Hombach J, Melegaro A, Edmunds JW, Beutels P: RH is a staff member of the World Health Organization. The views expressed in this article is that of the author and do not necessarily represent the views of the World Health Organization.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1741-7015/11/71/prepub"} +{"text": "The comprehensive evaluation, assessment, and management of patients with non-tooth-related conditions of the head and neck area are essential aspects of the practice of dental medicine. The manuscripts selected for publication in this special issue serve to illustrate the importance of close cooperation between oral and maxillofacial pathology, radiology, oral medicine, and head and neck anatomy in both the initial diagnostic and subsequent treatment phases when evaluating and treating patients with non-tooth-related conditions of the oral and maxillofacial complex.I would like to genuinely thank my Guest Editors, Dr. Neil S. Norton from Creighton University in Omaha, Neb, USA Dr. Preetha P. Kanjirath, from the University of Michigan at Ann Arbor, Mich, USA and Dr. Tarnjit Saini, Brooke Army Medical Center Fort Sam Houston, in San Antonio, Tex, USA for their assistance. Without their involvement and thoughtful discussions, this special issue would not have been possible. I also extend my thanks to the authors who have contributed to this special issue, as well as to the many reviewers who graciously volunteered with the peer-review process. Bone Diseases of the Jaws\u201d, by P. J. Slootweg, provides an overview of the more common and/or important lesions occurring in the oral and maxillofacial complex, while emphasizing the considerable overlap in clinical, histological, and radiological features among these entities.In the lead article in this special edition, \u201cY. Morimoto and colleagues review the usefulness of ultrasound imaging for the detection of noninvasive and soft tissue-related diseases and introduce three new potential applications of ultrasonography: guided fine-needle aspiration, measurement of tongue cancer thickness, and diagnosis of metastasis to cervical lymph nodes.Subsequent manuscripts explore the relationship between craniofacial pathology and anatomy. L. Sonnesen summarizes recent studies on the link between morphological deviations of the cervical vertebral column and craniofacial morphology, while Guest Coeditor Neil S. Norton and colleagues employ volumetric tomography to review the anatomy of the greater palatine canal and also to rule out a statistically significant association between the prevalence of maxillary sinus disease and the presence of concha bullosa and/or nasal septal deviation.R. A. Mesquita and colleagues critically review the available literature on the nonsurgical treatment of oral leukoplakia, while E. de S. Tolentino and colleagues present a well-documented case of an ameloblastic fibroma that illustrates the need to integrate radiology, oral and maxillofacial pathology, and head and neck anatomy in both the initial diagnosis and subsequent treatment of lesions of the maxillofacial complex.On behalf of my Guest Coeditors and myself, I hope that you will find the manuscripts that comprise this special issue both interesting and informative.Paul C. EdwardsPaul C. EdwardsPreetha P. KanjirathPreetha P. KanjirathTarnjit SainiTarnjit SainiNeil S. NortonNeil S. Norton"} +{"text": "There was an error in the Funding statement. The correct version of the statement is available below:YIB-M acknowledges the PDCB of CCG-UNAM and her support by a PhD fellowship (228320/210360) from CONACyT-M\u00e9xico. This research was partially supported by CCG-UNAM, by grant CB-103686 from CONACYT to JC-V, and grants 5R01GM071962-08 to JC-V and RO1GM030054-23 to MAS from the National Institute of General Medical Sciences, National Institutes of Health, USA. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of General Medical Sciences or the National Institutes of Health. EP-R was partially supported by DGAPA grant IN209511. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Juan Ignacio Arredondo-Jimenez was not included in the author byline. He should be listed as the eighth author and affiliated with National Center for Preventive Programs and Disease Control, Mexico, D.F. The contributions of this author are as follows: Contributed reagents/materials/analysis tools, Other: designed the method of calculating spatial risk areas."} +{"text": "Taenia pisiformis, also known as Cysticercus pisiformis, is the causative agent of cysticercosis and the cause of severe health problems in rabbits that negatively impacts on husbandry production. To date, there is no fast detection method to identify early infections in rabbits. In the present study, a new dot-ELISA-based on an endogenous antigen fatty acid-binding protein (FABP) was developed for the detection of cysticercosis, and its potential was then evaluated using test serum samples. Immunolocalisation showed that T. pisiformis FABP (TpFABP) localised to the parenchyma of the bladder wall of the cysticercus and perinuclear cytoplasm of parenchyma of the adult parasite. After cloning and expression, recombinant TpFABP (rTpFABP) protein was used for serodiagnosis of T. pisiformis infection in rabbits by dot-ELISA. The antibody was detected 14 days post-infection in rabbits experimentally infected with T. pisiformis. Based on the necropsy results, the sensitivity and specificity of 169 serum samples tested by rTpFABP dot-ELISA were found to be 98.2% (54/55) and 92.1% (105/114), respectively. These data suggest that the dot-ELISA developed in this study has potential for detection of T. pisiformis infection in rabbits.The larval stage of Taenia pisiformis iTpFABP is a suitable diagnostic antigen, and the results of study demonstrate the efficacy of the FABP-based dot-ELISA for potential detection of T. pisiformis cysticercosis in rabbit.Together, the data shows that r"} +{"text": "As well as biomedical risk factors, psychological factors have been reported to be related to mortality rate. The purpose of this study was to examine the relationship between life satisfaction and mortality in elderly people through an 11.8-year follow-up study of a prospective cohort.Among 3,600 participants of the Kangwha Cohort Study who survived in 1994, 1,939 respondents of the Life Satisfaction Index (LSI)-A questionnaire were included . The mortality risk for the period up to December 2005 was measured using the Cox Proportional Hazard Model.When the relationship between LSI and mortality was evaluated in men, the unsatisfied group with lower LSI scores showed a significantly higher risk of all-cause mortality than the satisfied group with higher LSI scores. In women, the unsatisfied group showed a significantly higher risk of all-cause mortality and cardiovascular mortality than the satisfied group.We found that elderly people with a lower LSI score, regardless of gender, were at risk of increased mortality from all causes, and low LSI score was also associated with cardiovascular mortality. Psychological factors and well-being, with no physical risks involved, have been reported to be associated with mortality rates -5. Two FIn a Finnish prospective study, life satisfaction was associated with a risk of total death in men, but only injury death in women . The FinThe primary survey for the Kangwha Cohort was conducted in March 1985 ,11. KangThe investigation team interviewed each subject using a structured questionnaire for demographic characteristics, including age, sex, education, chronic disease conditions, health behaviors (smoking and drinking), MMSE, ADL, IADL, and LSI. Life satisfaction was assessed by the LSI-A index, which was first prepared, validated, and published by Neugarten et al. . The intDisability in ADL was evaluated by the following criteria: bathing or showering, dressing, going to the toilet, transferring (in and out of bed or chair), and eating . For theThe outcomes of this research were cause-specific deaths. Data on deaths and their causes from March 1, 1994 to December 31, 2005 were obtained from the Statistics on the Causes of Death in Korea by the Korea National Statistical Office. The International Classification of Diseases, 10th Revision was applied to define the causes of death .LSI scores were grouped into tertiles using the dataset for men and women (N = 1939) to represent 'satisfied' (26-40 points), 'intermediate' (16-25 points), and 'unsatisfied' (0-15 points). Analysis of variance (ANOVA) and chi-square test were used to analyze the statistical differences among the characteristics of the study participants. Because there could be a variation in mortality risk by gender, a stratified analysis was made for men and women.p < 0.05 was used for all tests. Analyses were performed with SAS Windows version 9.1.We used the Cox proportional hazard model to test the relationship between LSI at baseline and subsequent risk of total mortality. Three models were set up to adjust for confounding variables. In Model I, age as a continuous variable was adjusted for. In Model II, in addition to age, the history of chronic disease , smoking habits , alcohol consumption, BMI (as a continuous variable), and education status were adjusted for. Smoking status and education level were included in the models as dummy variables. In Model III, IADL was further adjusted for. Hazard ratios (HRs) and 95% confidence intervals (CIs) were expressed for the results. A significance level of p < 0.01) (Table p < 0.01), had lower cognitive function, and had more disability in ADL and IADL. In women, although the outcome for IADL was similar to that in men, ADL, cognitive function, and education status were not related with LSI (Table General characteristics of men and women by LSI level are presented in Tables SI Table . HoweverTable Table In this prospective cohort study of Korean elderly subjects, low life satisfaction was associated with long-term mortality. It was also associated with cardiovascular disease-related mortality. These data are largely consistent with the findings of previous studies, such as a Finnish prospective study , a FinniThe present study used the LSI-A, a 20-item questionnaire, as an assessment tool of quality of life satisfaction. The Taiwanese study also used the LSI-A, and it is known to be useful in assessing overall quality of life satisfaction . PreviouThis study has several limitations. First, we adjusted BMI as a covariate. However, other classical biological risk factors for cardiovascular-related death, such as hypertension, diabetes, and dyslipidemia, were not included in this study. Second, non-inclusion of variables, such as depressive symptoms, sense of well-being, and social support, may be a limitation, although well-being is a dimension measured by the LSI-A in a comprehensive sense. Third, deaths from diseases other than cardiovascular disease, cancer, and all causes could not be analyzed due to an insufficient number of cases. More causes of death need to be analyzed in further studies. Fourth, selection bias is often an issue in studies of life satisfaction. The subjects in this study were an elderly group residing in the same administrative and geographical community, and approximately 85% of them engaged in agriculture, thus there was a great social and cultural similarity among them.In conclusion, our findings indicate an association between low life satisfaction and long-term mortality among elderly people, in particular cardiovascular disease mortality. Further studies are necessary to describe the relationship between life satisfaction and mortality risk for a wider range of diseases.ADL: Activities of Daily Living; BMI: Body mass index; CI: Confidence interval; HR: Hazard ratio; IADL: Instrumental Activities of Daily Living; LSI: Life Satisfaction Index; MMSE: Mini-mental state examination.The authors declare that they have no competing interests.All authors participated in drafting of the manuscript. HK performed the analysis and interpretation of data. JWS participated in planning the design and interpretation of data. BG and SWY participated in data analyses and interpretation. HO conceived the idea of the study, planned the design, collected data, and participated in interpretation of the data. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/12/54/prepub"} +{"text": "Organisation of health care for frail elderly is no longer adequate. Their complex requirements within the domains of prevention, care, cure, welfare and residence demand a new care model. Partitions need to be broken down and care-providers should work together on a single, integral basis, from diagnosis to treatment. This type of integrated care model has recently been implemented in the region of Walcheren in the Netherlands.Frail elderly living at home are preventively detected with the Groningen Frailty Indicator and their care demands are assessed with the EASYcare-instrument. Geriatric nurse practitioners and secondary care geriatric nursing specialists become supervisors of the care plans and care coordinators, whereas the G.P. is appointed as director and partner in prevention and care. The entire process is encompassed in multidisciplinary protocols, files (web-based), and multidisciplinary meetings and consultation within and between primary, secondary and tertiary care.The present study evaluates this integrated care model by examining its effects on frail elderly. Effects of the integrated care model on frail elderly are assessed for ADL-functions, experienced health, mental well-being, social functioning, quality of life and satisfaction with the provision of care. Furthermore, the effect on the use of care is explored. Validated instruments, such as the Katz-15 and Rand-36 are used.The design of this study is quasi-experimental: approximately 200 frail elderly patients of six G.P. practices that have implemented the integrated care model are compared with 200 frail elderly patients from six G.P. practices that have provided regular care. For this study, baseline measures will be compared with a three-month and a one-year follow-up. At the conference, the results of the three-month follow-up will be presented."} +{"text": "The Foot Function Index (FFI) is a self-report, foot-specific instrument measuring pain and disability and has been widely used to measure foot health for over twenty years. A revised FFI (FFI-R) was developed in response to criticism of the FFI. The purpose of this review was to assess the uses of FFI and FFI-R as were reported in medical and surgical literature and address the suggestions found in the literature to improve the metrics of FFI-R.A systematic literature search of PubMed/Medline and Embase databases from October 1991 through December 2010 comprised the main sources of literature. To enrich the bibliography, the search was extended to BioMedLib and Scopus search engines and manual search methods. Search terms included FFI, FFI scores, FFI-R. Requirements included abstracts/full length articles, English-language publications, and articles containing the term \"foot complaints/problems.\" Articles selected were scrutinized; EBM abstracted data from literature and collected into tables designed for this review. EBM analyzed tables, KJC, JM, RMS reviewed and confirmed table contents. KJC and JM reanalyzed the original database of FFI-R to improve metrics.Seventy-eight articles qualified for this review, abstracts were compiled into 12 tables. FFI and FFI-R were used in studies of foot and ankle disorders in 4700 people worldwide. FFI Full scale or the Subscales and FFI-R were used as outcome measures in various studies; new instruments were developed based on FFI subscales. FFI Full scale was adapted/translated into other cultures. FFI and FFI-R psychometric properties are reported in this review. Reanalysis of FFI-R subscales' confirmed unidimensionality, and the FFI-R questionnaires' response categories were edited into four responses for ease of use.This review was limited to articles published in English in the past twenty years. FFI is used extensively worldwide; this instrument pioneered a quantifiable measure of foot health, and thus has shifted the paradigm of outcome measure to subjective, patient-centered, valid, reliable and responsive hard data endpoints. Edited FFI-R into four response categories will enhance its user friendliness for measuring foot health. Foot problems commonly arise during our daily living activities,2. The pIn 1991, the Foot Function Index (FFI) was developed as a self-reporting measure that assesses multiple dimensions of foot function on the basis of patient-centered values. The FFI consists of 23 items divided into 3 subscales that quantify the impact of foot pathology on pain, disability, and activity limitation in patients with RA. The FFIIn 2006, the FFI was revised (the FFI-R) on the basis of criticisms from researchers and clinicians; items were added, including a scale to measure psychosocial activities and quality of life related to foot health.A literature review was conducted to develop a theoretical model of foot functioning, based oThe FFI-R in its current form is one of the most comprehensive instruments available. However, in a review article, questioOur aim is to assess the contribution of the FFI and FFI-R to the measurement of foot health in the fields of rheumatology, podiatry, and orthopedic medicine. This assessment should enable us to reflect on and improve the quality of the measure. Therefore, we conducted a systematic review of literature pertaining to the FFI and FFI-R that has been published in the English language from October 1991 through December 2010. The objectives were to: (i), Assess the prevalence of uses of the FFI and FFI-R in clinical studies of foot and ankle disorders; (ii), Describe the utility and clinimetric properties of the FFI and FFI-R as they have been applied in various clinical and research settings; (iii), Enumerate the strengths and weaknesses of the FFI and FFI-R as reported in the literature; (iv), Address the suggestions found in the literature for improving the FFI-R metrics.This study was about a systematic review of articles in which the FFI and/or FFI-R were used as measures of a variety of foot and ankle problems. Relevant studies were identified by English language publication searches of the electronic bibliographic databases Pub Med/MEDLINE, EMBASE, BioMedLib and Scopus from October 1991 through December 2010.foot function index, FFI scores, foot function index scores, and foot function index revised (FFI-R).were used as search terms and was applied to all databases. FFI instruments/measure and/or FFI-R instruments/measure had to be mentioned in the abstracts and in the full articles to be collected for in-depth scrutiny. Articles fulfilling the inclusion criteria were selected for the review. The article criteria included: (i) the words foot function index/FFI or revised foot function index/FFI-R in its reports/measures; (ii) full-length articles; (iii) written in English and published from October 1991 through December 2010; (iv) the study population described needed to have foot complaint(s)/problems; and (v) regardless of the country conducting the study, the full-length article must have been published in English or in a foreign language with the abstract in English.The key words: Selected articles that fulfilled the criteria were independently reviewed and collected by the authors to address the objectives and organize collected data into several tables.We created four tables to address the first objective of describing the measurement\u2019s uses Tables, and4.We designed a data-collection form to address the second objective. This form was assessed in a pilot study by collecting data from ten articles out of the collection of qualified articles; it was revised before being used in its current format. The variables used in this data-collection form were: (i) the instrument and year the article was published; (ii) the first author\u2019s name; (iii) the objectives of the study; (iv) the population characteristics, sample size, and diagnosis; (v) psychometric analysis ; (vi) items/domains/subscales of the FFI or FFI-R used in the study; (vii) response type; and, (viii) a short summary evaluation of each study. Therefore, this data form recorded the analytic statements extracted from each article, and 6 tables were created Tables, and10.This was a qualitative summary of the results as found in TableTableQuantitative data were reported using simple statistics expressed as the sum, means, and standard deviations for continuous variables and as frequencies for categorical data. Tables, and4 ATo address specific critiques of the FFI-R found in the literature, the unidimensionality of the FFI-R and its subscales were evaluated against the Rasch model. The statistical package Winsteps version 3.72.3 was usedReliability was estimated with Cronbach\u2019s Alpha and Rasch person reliability statistics. Both indices reflect the proportion of variance of the person scores or measures to total variance . Unlike Cronbach\u2019s Alpha, Rasch person reliability is based on the estimated locations of persons along the measurement continuum, excluding those with measures reflecting extreme (zero or perfect) scores and including cases with missing data. For both indices, our criterion for acceptability was .80.One requirement of the Rasch model is monotonicity: the requirement that, as person ability increases, the item step response function increases monotonically. This meArticles were obtained by using the search method defined in the Methods section; the search results included 752 articles from PubMed/MEDLINE and 640 articles from Embase. Further screening and selection procedures, as detailed in FigureAmong the 78 studies, we identified 4714 study participants for whom the FFI or FFI-R instrument had been used to measure foot health. This sample consisted of 1914 (41%) male participants and 2688 (57%) female participants, with a mean age of 48.58 years . There was a discrepancy of 2% between the sums of male and female participants, because gender was not reported in three studies Table.TableIn summary, the FFI with all 3 domains, or as subscales, was frequently chosen as a measurement instrument across various studies and countries and among various age groups and sexes, for the assessment of acute and chronic foot and ankle conditions.The uses of the FFI and FFI-R are provided in detail in TablesCategory A New Instruments. Includes four articles in which foot health measures are described including the original FFI[Category B FFI as Criterion Validity. Articles in this category describe several health measures and use the FFI full scale or subscales to validate these measures. Bal et al.[Category C Cultural Adaptation or Translation. The first translation of the FFI was the Dutch-language instrument known as Dutch FFI-5pts[Tableinal FFI, the FFIinal FFI. The FFIinal FFI. The Ankinal FFI; measureinal FFI modifiedl et al. found a l et al. found thl et al. measuredl et al. developel et al. reportedl et al.. These vl et al. for the l et al. for the FFI-5pts. The GerFFI-5pts; the FFIFFI-5pts, Taiwan FFI-5pts, TurkishFFI-5pts, and CzeFFI-5pts. There wFFI-5pts. These tTableThe FFI is one of the outcome measures most frequently used by AOFAS members. It was TableThe FFI also was used to measure foot health outcomes associated with medical interventions Table, such asInvestigators had chosen the FFI scores or the subscale scores to determine the prevalence and disease burden of foot and ankle conditions in the general population Table. Novak eIn all, we found the FFI instrument was frequently chosen as an outcome measure of surgical, orthotic, and medical treatments, but its application was wider than we originally imagined. It was not limited to outcome measures; FFI scores were also applied in the promotion of foot health as a common public health issue and in increasing the awareness of health system administrators. The FFI was also used in the validation of newly developed foot health measures.FFI: The FFI questionnaire had good psychometric properties[operties-100, andoperties. In a stoperties reportedoperties. All theoperties.There are weaknesses of the FFI. During the development of the index, clinicians generated the questionnaire items without patient participation,97; therFFI-R: The FFI-R was developed in response to criticism of the FFI and to address issues of contemporary interest. Most original items from the FFI were selected in the development of FFI-R, and new items about footwear and psychosocial factors were added, which improved its construct coverage. Patients and clinicians were involved in the generation of items. Its design closely followed the ICF theoretical model[al model; its psyal model, allowinal model,76 did mFor the FFI-R L (68 items), person All subscales of the FFI-R had strong person reliability estimates Table, rangingThe response category analyses for each of the subscales (done after collapsing Categories 5 and 6) revealed that, for the first three subscales , the response categories behaved as required by the Rasch model. However, for the subscales of limitation and social issues , there was some indication that respondents had difficulty distinguishing between, \u201c2 = A little of the time,\u201d and, \u201c3 = Some of the time.\u201d We considered, then, collapsing these categories and making all FFI-R subscales have four possible response categories. This would ensure uniformity of the measure and decrease the burden on patient response. Therefore, the first three subscales, which measure severity, \u201c3 = Severe pain,\u201d \u201c4 = Very severe pain,\u201d and \u201c5 = Worst pain imaginable,\u201d were collapsed. This was justified because all three captured the notion of severe pain. Overall, the analyses showed that the response to each item functioned well with the four-item response categories.This review evaluated 78 eligible articles Figure. In the In recent articles about FFI used as outcome measures, the authors have included the clinical measures; the effect size, and standard response mean, and minOur literature search was limited to publications written in the English language and covered only publications until 2010; therefore, this might exclude the FFI- and FFI-R\u2013related published articles not written in English, as well as those more recent articles published in English.The FFI pioneered measuring outcomes in foot health. This instrument has been tested through time and adapted in its measures as it was frequently used in full scales or subscales to measure outcomes in various clinical practice or research studies. The FFI has also had a role in shifting the paradigm from a reliance on physical and biochemical findings as outcomes to the use of outcomes that are relevant to patients. Thus, the measure established patient-centered, valid, reliable, and responsive hard data endpoints. The rating scales also underwent changes; for practicality and user-friendliness in clinical and research settings. The FFI was recognized as a valid instrument and used as a validation criterion of other measures. It was adapted and translated into multiple languages. It was applied to all age groups, across genders and was useful in measuring varied medical and surgical conditions.In realizing the scope of FFI applications, we acknowledge the contributions of friends and colleagues around the world who not only used the FFI in their studies but also made adaptations and translations to make the FFI a versatile instrument in promoting and maintaining foot health. The FFI-R has good psychometric properties and is available in long and short forms for ease of clinical use. In response to findings in this review, we conducted a rigorous analysis to strengthen the metrics of the FFI-R and changed the rating scales to be more user-friendly and practical.Both the FFI and FFI-R are in the public domain and permission to use them is free of charge. They are available from the developers of these instruments and from the AOFAS web site. These instruments are self-administered and are written at an eighth-grade reading level. The FFI scores are interpreted as 0%-100% for each subscale and the overall score. Higher FFI and FFI-R scores indicate poor foot health and poor foot health-related quality of life. The FFI and FFI-R put minimal burden on respondents and the questionnaires are not emotionally sensitive. The administrative burden is also minimal and it does not require formal training to score or to interpret. TranslaThis review attests to the widespread use of foot health measures, and we have noticed the advancement of foot health in general across diagnoses. It has been a privilege for us to serve patients, clinicians, and researchers to fulfill the mission in improving foot health through the use of the FFI and FFI-R. These instruments are available for users, and can be downloaded as they are presented as electronic files.AOFAS: American Orthopedic Foot and Ankle Society; CTT: Classical test theory; EMBASE: Excerpta Medica Database; FFI: Foot Function Index; FFI-R: Foot Function Index Revised; EBM: Elly Budiman-Mak; FFI-R L: Foot Function Index Revised Long Form; FFI-R S: Foot Function Index Revised Short Form; HAQ: Health Assessment Questionnaire; IRT: item response theory; JM: Jessica Massa; KJC: Kendon J Conrad; Medline: Medical Literature Analysis and Retrieval System; PUBMED: public Medline; RA: rheumatoid arthritis; RMS: Rodney M. Stuck; VAS: visual analog rating scale; AAOS: American Academy of Orthopedic Surgeon; ANOVA: Analysis of Variance; AOS: Ankle Osteoarthritis Index; BMD: Bone Mineral Density; CA: Crohnbach\u2019s Alpha; CRI: Clinical Rating Index; CV: Calcaneal Varus; DAS 44: Disease Activity Score in 44 joints of patient with rheumatoid arthritis (RA); DX: Diagnosis; EF: External Fixation Procedure; ES: Effect Size; FAAM: Foot and Ankle Ability Measure; FFI-5pts: Dutch Foot Function Index with 5 point Likert Scale; FFI-G: Foot Function Index - German Language; FHSQ: Foot Health Status Questionnaire; FIS: Foot Impact Scale; FPS: Foot Problem Score; FSI: Foot Structure Index; FX: Fracture; HFS: Hind Foot Function Scale; HMIP: Hallux Metatarso-interphalangeal Joint; HR: Hallux Rigidus; ICC: Interclass Correlation Coefficient; JIA: Juvenile Idiopathic Arthritis; JRA: Juvenile Rheumatoid Arthritis; LMIP: Lesser Metatarso-interphalangeal Joint; MCS: Mental Component Score of SF-36; MDC: Minimal Detectible Change; MFA: Musculoskeletal Function Assessment; MFDQ: Manchester Foot Disability Questionnaires; MID: Minimal Important Difference; MODEMS: Musculo-skeletal Outcome Data Evaluation and Management System; MTP: Metatarsophalangeal Joint; NA: Not Applicable; OA: Osteoarthritis; PAS: Physical Activity Scale; PCS: Physical Component Score of SF-36; PedQL: Pediatric Quality of Life Scale; PF: Plantar Fasciitis; PTTD: Posterior Tibialis Tendon Dysfunction; QOL -12: Quality of Life 12 items short form; RAI: Ritchie Articular Index; RCT: Randomized Control Trial; SD: Standard Deviation; SF-36: Rand 36 items health survey form; SF-36 MCS: Mental Component Score of SF-36; SF-36 PCS: Physical Component Score of SF-36; SF-12: Rand 12 items short form health survey; SFC: Steinbrocker Functional Class; SMFA: Musculoskeletal Function Assessment; SRM: Standard Response Mean; SI: Stroke Index; TAA: Total Ankle Arthroplasty; TMT: Tarso Meta-metatarso Joint; UCLA: University of California - Los Angeles; WOMAC: Western Ontario MacMaster University Osteo Arthritis Index.The authors declare that they have no competing interests.EBM, KJC, have contributed in drawing the concept and design of this paper, EBM initiated the literature search, reviewed, scrutinized them, and collected the abstracts and organized into tables. KJC, RMS and JM reviewed the tables and all authors participated in drafting the manuscript. KJC and JM also reanalyzed the original FFI-R data and revised the subscales and FFI-R response categories. All authors participated in revising the manuscript and have given final approval of the version to be published.Revised FOOT FUNCTION INDEX (FFI-R).Click here for fileRevised FOOT FUNCTION INDEX (FFI-R) Short Form. Click here for file"} +{"text": "It brought together 96 scientists from the Asian region and beyond to exchange ideas, report on progress, make a gap analysis, and distill prioritizing settings with a focus on the Asian region. Key objectives of this international symposium were to agree on solutions to accelerate progress towards decreasing transmission, and human mortality and morbidity caused by the three major cestode zoonoses ; to critically assess the potential to control these diseases; to establish a research and validation agenda on existing and new approaches; and to report on novel tools for the study and control of cestode zoonoses.Globally, cestode zoonoses cause serious public health problems, particularly in Asia. Among all neglected zoonotic diseases, cestode zoonoses account for over 75% of global disability adjusted life years (DALYs) lost. An international symposium on cestode zoonoses research and control was held in Shanghai, China between 28 Please see Additional file Cestode zoonoses are emerging, re-emerging or spreading worldwide, and are classed as neglected infectious diseases or neglected tropical diseases (NTDs), and neglected zoonotic diseases (NZDs) ,2. AmongTo respond to the increased requirement for strengthening international collaborations on research and control of cestode zoonoses with a focus on the Asian region, an international symposium was jointly proposed and co-organized in Shanghai by the National Institute of Parasitic Diseases, part of the Chinese Center for Disease Control and Prevention, and the Asahikawa Medical University in Japan. A total of 96 scientists attended and engaged in topics as diverse as treatment, diagnosis, molecular biology, epidemiology, transmission ecology, public health and health policy, and progress in research and control of cestode zoonoses in Asia, including gap analysis and priority settings.This international conference focused on the control of cestode zoonoses, and involved 96 participants from 11 countries, the Chinese Ministry of Health (MOH), the World Health Organization (WHO) Regional Office and non-government organizations (NGOs). During the two-day symposium, five keynote speakers introduced the current global situation of cestode zoonoses from various perspectives. Thirteen speakers addressed the Asian status of cestode zoonoses, and recent achievements on biological, spatial, molecular and diagnostic aspects. Twenty-two participants in four parallel sessions shared their experiences and progress on epidemiology, ecology, biology, immunology, molecular taxonomy and phylogeny, control strategy, diagnosis, vaccine research, clinical treatment, and public policy related to cestode zoonoses control the current endemic/epidemic situation of cestode zoonoses with a focus on Asian countries; b) the current national and international control strategies for cestode zoonoses; c) effective approaches to implement intersectoral efforts on survey and control of cestode zoonoses; d) importance of epidemic surveillance networks and information reporting systems; and e) opportunities, challenges, feasibilities, sustainability and technique demand for cestode zoonoses control.During the symposium, the Delphi study was applied for obtaining and understanding control bottleneck, research and resource gaps, priority settings for research and control, and other related problems from all domestic and foreign participants . InformaThe main contributions were in the following areas: biology and eco-epidemiology, control and interventions, molecular biology and diagnostics, gap analysis, and research priorities.Echinococcus spp were reported as high as 61.7-64.5% in Tibetan fox populations [Echinococcus spp in small mammals on the Tibetan plateau was also conducted by morphology and hemi-nested polymerase chain reaction (PCR) with a prevalence of 7.8%, in which 44 of 45 samples were identified by the hemi-nested PCR as infections of E. multilocularis (97.8%) and one considered as E. granulosus in Microtus fuscus[E. multilocularis, E. sibiricensis and E. russicensis[Echinococcus species to be conducted. However, some of the above-mentioned lesions have been confirmed to be an intra-species variation of E. multilocularis by the most recent molecular studies [According to recent surveys with a focus on epidemiology in wildlife, prevalences of ulations . A surveus fuscus. Three mssicensis. These s studies ,19. The E. multilocularis is more intense in distinct ecological systems with various intermediate host communities, landscape, and climates in continental Asia [From continental to regional scales, human AE spatial distribution appears to be highly aggregated and forms discrete patches of endemicity within which hotspots of much larger prevalences may occur. Using regional spatial models helps to explain why transmission of tal Asia -25. The tal Asia . Due to tal Asia .E. multilocularis, and thus might help in the design of evidence-based monitoring and control programs. Some recent investigations in the Asian continent were introduced in the symposium with the perspective of featuring distinct types of transmission ecosystems based on intermediate host communities. These can serve as a reference for further in-depth research and help when considering surveillance systems are being considered [The analysis of spatial patterns of human and animal case distribution can provide information on transmission processes of nsidered ,27,28.Taenia species were co-endemic in farming areas, where neurocysticercosis is an emerging public health concern in China and Central Asian Countries, and an important domestic host of E. multilocularis. However, dog-centered hydatid control interventions in rural pastoral communities are more difficult where CE and AE are co-endemic. Many questions need answering such as the natural re-infection rate in dogs for E. granulosus and E. multilocularis, and the potential for dogs to maintain fox-independent transmission of E. multilocularis[Dogs have been recognized as the main definitive hosts of locularis,32. In tlocularis.Regarding activities aimed at raising awareness and thus being instrumental in the disease\u2019s control, it was reported that a series of target-specific health educational materials were developed and applied for community-based health education in the Qinghai-Tibet plateau region with the overall aim of raising compliance of Tibetan populations in echinococcosis endemic areas. The materials printed consisted of a Tibetan calendar, a notebook, a pencil bag, a teaching wall chart, a school health textbook, a picture storybook, an animated cartoon and a picture poster, and these were each targeted for the different population needs of religious staff, local government staff, students, and residents in a CE patient with multi-infected organs was reported, in which the treatment was a cyclic combination therapy of ABZ at 400\u00a0mg twice a day for five days, with PZQ at 800\u00a0mg a day for two more days of the week. The regiment was repeated for 10 courses with a two-week interval between courses. The results indicated that both lung and liver cysts reduced in size and no side effects were observed, suggesting a potential additional option for medical therapy . To explTo meet reasonable medical service needs of remote populations from different ethnic groups, a form of teleconsultation was introduced as a novel approach for remote clinical consultation, multi-disciplinary discussion, surgery demonstration, and theoretical training. Echinococcosis patients in remote areas could thus, in theory, benefit from optimal implementation of local treatment, correct referral, as well as high-quality, inexpensive and convenient medical consultation.Taenia solium taeniasis, and consequently the risk of cysticercosis, the usefulness of traditional oral anthelmintic (consisting of pumpkin seeds combined with areca nut extract) was assessed in a community-based treatment for human taeniasis. The results showed that the traditional Chinese herbal treatment was highly effective in expelling intact tapeworms in over 89% of taeniasis cases [T. solium transmission in the future. Theoretically at least, T. solium cysticercosis is capable of being eradicated entirely. More recently, treatment of pigs with oxfendazole and development of an extremely effective vaccine for pigs (Tsol18) have been shown to have great potential in reducing transmission at the community level [To control is cases . Other ety level , particuty level . HoweverEchinococcus variants, genotypes or strains, some of which are poorly or not infective to humans, a molecular taxonomic discussion sought to reinforce the necessity to revise the taxonomic status and to recognize nine species in the genus Echinococcus i.e. E. granulosus, E. equinus, E. ortleppi, E. canadensis, E. felidis, E. multilocularis, E. shiquicus, E. vogeli, and E. oligarthrus[E. canadensis, the previous genotypes of E. granulosus i.e. G6, G7, G8, and G10 should now be regarded as E. canadensis[E. canadensis (G6) was detected in a human infection case. E. granulosus was not only found in humans and sheep, but also reported in a cat, while three genotypes of E. multilocularis \u2013 the Mongolian, Asian, and North-American types \u2013 were confirmed from a variety of hosts including the Asian type in human cases [Based on the need for reappraisal of the species within the genus igarthrus-39. Accuan cases ,41.E. multilocularis was recommended as a good candidate for the serological detection of almost all active AE cases (PNM classification) [E. granulosus was reported to be another good candidate to detect the majority of active CE cases i.e. CE2, CE3a, and CE3b (WHO-IWGE US classification) [Echinococcus, EgAgB8/3, was assessed for coproantigen detection in an ELISA test for dog infection, revealing 85% sensitivity and 95.7% specificity [E. granulosus in dogs was performed and a series of proteins associated with survival, development, movement and modulation were identified, and thus improved the repertoire of candidate proteins for vaccination, immunodiagnosis, and drug development [Echinococcus species in carnivore feces, which is useful for large-scale as well as fine spatial resolution studies, was presented [E. granulosus and showed that Th1 is the major response in the early stage of infection, while Th2 is the major response between the 8th and 16th week after the infection [Recombinant Em18 from ication) and was ication) ,44. Rapiication) ,46. Alsocificity ,48. Rececificity . A proteelopment . Vaccineelopment . To overresented . Some reT. solium using mitochondrial gene markers, it was reported that there are two genetically distinct geographic subgroups: Asian and African-American [Taenia asiatica, molecular analysis strongly indicates that T. saginata and T. asiatica were separated into two independent species. On the other hand, hybridization between these two human Taenia species seems to be an ongoing event in the co-endemic areas [Molecular phylogeography has highlighted the evolution and dispersal history of zoonotic taeniid cestodes. Based on the phylogeographic analyses for American ,53. Althic areas -56.T. solium between humans and animals, and in optimizing baseline studies and intervention follow-ups and monitoring.A review of recent advances in immunodiagnosis of cysticercosis and some latest research progress were reported including development of an immunochromatographic test, and a simple and reliable purification method of glycoproteins for immunodiagnosis of human and pig cysticercosis . A reliaIt is recognized that substantial gaps exist in the control and research of cestode zoonoses globally, particularly in Asia. Key challenges include capacity building; governance and quality control of research; identifying knowledge gaps of burden estimation of the diseases; development of socioeconomic indicators; resource mobilization; need to develop inter-programmatic, intersectoral prevention and control strategies; and linking research, program and policy for evidence-based decision making.E. granulosus and CE, and potential for control programs, should include country-wide mapping of cestode zoonoses, and the reporting of medical and veterinary data in a uniform way. As well as that, basic livestock information is required, for example: 1) Do cysts accumulate? 2) Does use of repeated low dose ABZ for G1 nematode infections have any effect on hydatid cysts?Epidemiology, ecology and surveillance of It is necessary to further improve copro-ELISA tests, to assess loop-mediated isothermal amplification (LAMP) and copro-DNA detection, and to run multi-center assessments of copro- and serological tests and their standardization. Up until now, low-cost serological tests for CE and livestock serology remain problematic. In control and prevention of CE, there is a need to: evaluate integrated use of EG95 vaccine and PZQ trials over a five-year period to measure dog re-infection rates in different endemic zones pre- and during PZQ dosing regimes, to minimize dog dosing frequencies that can be used in a given transmission zone/system, to develop setting-specific health education materials, and to further undertake research on experimental dog vaccines. There is also a growing interest in, and importance applied to, systematically analyzing and evaluating the four-five year progress data from dog and human indices for interim assessment of the Chinese National Control Program of Echinococcosis. In the program, mainly comprehensive control measures are taken including dog management and monthly deworming with PZQ, health education, livestock management and immunization, case chemotherapy with ABZ, and surgical treatment.E. multilocularis isolates ; copro-tests for parasite and host identification; evaluation of uncertainty in diagnostic tests; reliability of serological tests for early AE lesion detection in humans; re-infection rates in dogs; and potential for dogs to maintain a synanthropic cycle without significant fox populations. In control and prevention of AE, the gaps which need to be investigated are centered around: the role of domestic dogs being important in the transmission of E. multilocularis in co-endemic areas under hydatid control programs, the problem of stray dogs in some areas, the potential for sustainable dog-small mammal transmission, and the behavior of the red fox, the Tibetan fox and the corsac fox in the wild versus among human communities.In epidemiology, ecology and surveillance of AE, the gaps are: mapping distribution; development of landscape risk maps; age structure of small mammal host populations for infection studies; reliable rodent taxonomy for host species in Eurasia; assessment of the DNA profiles in unusual Inner Mongolian T. solium; multi-center assessment of current serological tests for human and pig cysticercosis; specificity in relation to T. hydatigena; effective DNA detection by LAMP for field use; and increased studies on epidemiology and transmission of T. solium in Asia. In control and prevention of taeniasis/cysticercosis, many factors need to be addressed such as the availability of copro-ELISA for mass screening, drugs for taeniasis mass drug administration , and integrated use of Tsol18 vaccine and oxfendazole in pig populations through pilot studies.Gaps in research on epidemiology, ecology, and surveillance of taeniasis/cysticercosis need to be focused on distribution maps for countries and accurate hospital data; DALYs calculation for cysticercosis burden, and proportion of epilepsy due to neurocysticercosis in China and the Southeast Asia; accurate porcine cysticercosis data; the minimum effort required to reduce transmission to Ro\u2009<\u20091; transmission dynamics basic parameters for Cestode zoonoses, as one group of neglected tropical diseases and neglected zoonotic diseases, mainly affect the poorest sectors of the populations and poor livestock keepers living in rural, often remote and disadvantaged regions of developing countries, and contribute to serious public health problems in the Asian region. China is one of the countries with the highest disease burden of cestode zoonoses in the world, where human CE, AE, and cysticercosis are endemic, and even co-endemic, locally. Human AE is known to be common in certain rural agricultural and pastoral communities. Though globally rare, about 91% of new AE cases globally occurring each year are detected in China with about 380 endemic counties and about 86 million people at risk, particularly in the Qinghai-Tibet plateau region and Western Sichuan. Cestode zoonoses are, however, neglected with little attention from policy-makers, lack of priority within health strategies, and inadequate baseline research.In this symposium, research priorities were recognized. They included the need to understand the relationship between infectious diseases and poverty, and contribute to priority settings for plans to control those diseases by the introduction of the \u201cone health, one world\u201d concept with trans-disciplinary approaches. It is essential to apply modern tools to obtain scientific evidence on parasite species, strains or isolates, as well as on the confirmation of the diseases themselves, and to further understand epidemiology and transmission ecology. The priorities were also raised to promote optimal opportunities and strategy for control and international collaborations with emphasis on those features, and to improve diagnosis, treatment and control. In addition, estimations of burden of diseases, development of socioeconomic indicators, developing a country-specific control strategy and implementing it, evidence-based interventions, and the use of identified best practices were all identified as important.Cestode zoonoses, as a neglected group of potentially life-threatening infectious diseases, pose an important public health challenge globally, particularly in Asian countries. They have substantial socioeconomic impacts as they impact groups beyond those directly affected, disproportionately impact on resource-poor communities in rural or remote areas, and furthermore, impact the health and productivity of livestock.At this international conference, it was agreed that the following initiatives and recommendations should be undertaken:\u2022Making a critical assessment on the potential for control of cestode zoonoses focusing on the regions where the populations are at higher risks;\u2022Establishing a research and validation agenda on new approaches, and novel tools for study and control of the diseases;\u2022Developing a work-plan of action targeting interventions; and\u2022Exploiting more resources, favorable public policy, and control options and strategies against cestode zoonoses.The WHO has recently listed echinococcosis as both a NZD and a NTIn this symposium, the possible next steps to achieve an integrated animal-human health approach have been recommended. These include:\u2022Promoting the concept of \u201cone health\u201d by the development of integrated \u201ccontrol packages\u201d to deal with health problems in people, livestock, and other domestic and wild animals;\u2022Taking effective measures to raise the profile of the NZDs both internationally and within affected countries;\u2022Systematically collecting data on the incidence of those zoonoses with support by studies to estimate their dual burden on people and on livestock, to quantify under-reporting, and to identify communities and groups at risk; and\u2022Investing in the development of new tools needed to effectively control these diseases, particularly in the field of diagnostics.It is high time to further understand these diseases and fill the gaps on what needs to be done, as well as to make joint efforts to control and research these diseases with a focus on the priority areas identified in this international conference. In addition, it is easy to define the research priorities, but translating these into operational control methods remains a big challenge. This could not be conducted effectively without a good health system and by increasing the prioritization of these diseases.ABZ: Albendazole; DALY: Disability adjusted life years; NTD: Neglected tropical disease; NZD: Neglected zoonotic disease; WHO: World Health Organization; CE: Cystic echinococcosis; AE: Alveolar echinococcosis; PZQ: Praziquantel; MOH: Ministry of Health; NGO: Non-government organization.The authors declare that they have no competing interests.NX and PSC wrote manuscript; NX, JWY, and WD collected, organized, and reviewed the data; NX, PG, PSC, and AI edited and revised the manuscript; and AI co-organized the symposium. All authors read and approved the final manuscript.NX: Professor, Deputy Director of National Institute of Parasitic Diseases, China CDC; WHO Collaborative Center for Malaria, Schistosomiasis and Filariasis; Key Laboratory of Parasite and Vector Biology, Ministry of Health, 207, Ruijin No. 2 Road, Shanghai 200025, P.R. China;JWY and WD: Assistant Researchers of the National Institute of Parasitic Diseases, China CDC; WHO Collaborative Center for Malaria, Schistosomiasis and Filariasis; Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai, P.R. China;PG: Professor of Ecology, Chrono-environment, University of Franche-Comt\u00e9/CNRS and Institut Universitaire de France, Besan\u00e7on, France;PSC: Professor of Biological Sciences and Director of Cestode Zoonoses Research Group, School of Environment and Life Sciences, University of Salford, Salford, Greater Manchester, M5 4WT, UK;AI: Emeritus and Visiting Professor of Department of Parasitology, Asahikawa Medical University, Midorigaoka Higashi, Asahikawa 078\u20138510, Hokkaido, Japan.Multilingual abstracts in the six official working languages of the United Nations.Click here for file"} +{"text": "The Canadian Hypertension Education Program (CHEP) started an ambitious dissemination and implementation program (D&I) in 1999 to 66% iCHEP\u2019s D&I program includes three components: dissemination, implementation and addressing barriers. Dissemination has been achieved through a passive-to-active dissemination process by publishing in multiple formats - peer-reviewed and non peer-reviewed - with content tailored to end users, including patients and their families. Another important aspect has been the development of tools to help professionals in daily decision making for the management of hypertension .Implementation happens when the information is used locally and barriers to the translation of such information are addressed. Characteristics of D&I programs usually include the following elements: multifaceted, multiple audiences, multimedia, consistent information and messages; sustainable, credible, using the appropriate language, and realistic and applicable. Each barrier is specific and should be addressed individually. A barrier can be, for example, related to access to professional services, diagnostic procedures, specific therapeutic procedures or different provincial/local regulations. Some barriers may be local or systemic or absence of structured care in the management of chronic diseases.In our experience, critical success factors for guidelines implementation are: a strong methodology for the development of high quality recommendations, an annual review of the scientific literature, and endorsement and participation of leading experts and key opinion leaders.Items listed on Table"} +{"text": "Fatigue is a common complaint among elementary and junior high school students, and is known to be associated with reduced academic performance. Recently, we demonstrated that fatigue was correlated with decreased cognitive function in these students. However, no studies have identified cognitive predictors of fatigue. Therefore, we attempted to determine independent cognitive predictors of fatigue in these students.kana pick-out test, semantic fluency test, figure copying test, digit span forward test, and symbol digit modalities test. The participants also completed computerized cognitive tests (tasks A to E on the modified advanced trail making test). These cognitive tests were used to evaluate motor- and information-processing speed, immediate and delayed memory function, auditory and visual attention, divided and switching attention, retrieval of learned material, and spatial construction. One year after the tests, a questionnaire about fatigue was administered to all the participants.We performed a prospective cohort study. One hundred and forty-two elementary and junior high school students without fatigue participated. They completed a variety of paper-and-pencil tests, including list learning and list recall tests, After the follow-up period, we confirmed 40 cases of fatigue among 118 students. In multivariate logistic regression analyses adjusted for grades and gender, poorer performance on visual information-processing speed and attention tasks was associated with increased risk of fatigue.Reduced visual information-processing speed and poor attention are independent predictors of fatigue in elementary and junior high school students. Fatigue refers to the feeling that people may experience after or during prolonged activity . FatigueWhen students proceed to junior high school from elementary school, a multitude of changes occur in their environment, which have the potential to cause a variety of behavioral and emotional problems . One exakana pick-out test) improved from elementary to junior high school. Thus, these cognitive functions develop from childhood to adolescence. Based on these findings, these cognitive tests were advantageous in the present study for evaluation of cognitive development in children and adolescents.Executive function is defined as a set of cognitive control processes that permit goal-directed behavior and that develop dramatically between childhood and adolescence . In studFatigue has been shown to be associated with impaired cognitive function in studies of adults -19. In cParticipants were enrolled from the 4th, 5th, and 6th grades in an elementary school and from 7th, 8th, and 9th grades in a junior high school in Hyogo Prefecture, Japan, between November 2006 and December 2006 . Most ofThe severity of fatigue was measured using the Chalder Fatigue Scale . The ChaStudents performed a variety of paper-and-pencil and computerized cognitive tests ,27. Partkana pick-out test and longer reaction time on task A of mATMT were associated with a higher risk of fatigue in the elementary and junior high school students. In addition, longer reaction times on task B showed a trend toward increasing risk of fatigue in these students . No other scores of cognitive tests were predictors of fatigue in these students.In order to identify cognitive predictors associated with fatigue in the elementary and junior high school students, univariate and multivariate logistic regression analyses were performed Table . Althougp = .019; Table In task A of mATMT, even after the subtraction of the reaction time associated with motor processing (reaction time on task D), multivariate logistic regression analyses showed that longer reaction time was a predictor of fatigue in the elementary and junior high school students [OR: 1.62, 95% CI: 1.08 to 2.43 (per 1 SD increase); These prospective data demonstrate that lower scores on the symbol digit modalities test and longer reaction times on tasks A and B of the mATMT were associated with the risk of fatigue in elementary and junior high school students. These findings were independent of grade and gender. To our knowledge, this prospective cohort study provides the first evidence identifying cognitive risk factors of fatigue in students.Tasks A and B of the mATMT primarily require visual information-processing speed and attention. In contrast, task C requires visual search rather than visual information-processing speed and attention. Although longer reaction times on tasks A and B were associated with increased risk of fatigue, longer reaction times on task C were not associated with increased risk of fatigue in the elementary and junior high school students. In addition, a lower score on the symbol digit modalities test, which was also used to assess visual information-processing speed and attention, was associated with increased risk of fatigue in these students. These results suggest that low visual information-processing speed and attention are predictors of fatigue in children and adolescents.Fatigue is defined as the difficulty in initiating or sustaining voluntary activities , suggestWe did not identify a mechanism by which low visual information-processing speed and attention increased the risk of fatigue. In patients with CFS , the relThe present study has two limitations. First, we performed this study with a limited number of participants. To generalize our results, studies involving a larger number of participants are essential. Second, we did not exclude developmental disabilities from analyses and did not assess the socioeconomic status and intelligence quotient.The present results provide evidence that low visual information-processing speed and attention are independent predictors of fatigue in elementary and junior high school students. Fatigue is associated with impairment of academic performance in elemeCCFS: Childhood chronic fatigue syndrome; CI: Confidence interval; mATMT: Modified advanced trail making test; OR: Odds ratio; RT: Reaction time; SD: Standard deviation.1Department of Physiology, Osaka City University Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka City, Osaka 545-8585, Japan, 2Molecular Probe Dynamics Laboratory, RIKEN Center for Molecular Imaging Science, 6-7-3 Minatojima-minamimachi, Chuo-ku, Kobe City, Hyogo 650-0047, Japan, 3Department of Medical Science on Fatigue, Osaka City University Graduate School of Medicine, 1-4-3 Asahimachi, Abeno-ku, Osaka City, Osaka 545-8585, Japan, 4Department of Clinical, Health and Special Needs Education Needs, Hyogo University of Teacher Education Graduate School of Education, 942-1 Shimokume, Kato City, Hyogo 673-1494, Japan.The authors declare that they have no competing interests.KM took part in the planning and designing of the experiment and cognitive tests, data analyses and manuscript preparation. MT contributed to the design and planning of the experiment and cognitive tests, data analyses and manuscript preparation. SF, EY and YS contributed to the design and planning of the experiment and in data preparation. KIM contributed to the design and planning of the experiment and recruited the participants. YW took part in the planning and design of the experiment and cognitive tests and helped write the manuscript. All authors read and approved the final manuscript."} +{"text": "There is global concern over significant threats from a wide variety of environmental hazards to which children face. Large-scale and long-term birth cohort studies are needed for better environmental management based on sound science. The primary objective of the Japan Environment and Children\u2019s Study (JECS), a nation-wide birth cohort study that started its recruitment in January 2011, is to elucidate environmental factors that affect children\u2019s health and development.Approximately 100,000 expecting mothers who live in designated study areas will be recruited over a 3-year period from January 2011. Participating children will be followed until they reach 13 years of age. Exposure to environmental factors will be assessed by chemical analyses of bio-specimens , household environment measurements, and computational simulations using monitoring data as well as questionnaires. JECS\u2019 priority outcomes include reproduction/pregnancy complications, congenital anomalies, neuropsychiatric disorders, immune system disorders, and metabolic/endocrine system disorders. Genetic factors, socioeconomic status, and lifestyle factors will also be examined as covariates and potential confounders. To maximize representativeness, we adopted provider-mediated community-based recruitment.Through JECS, chemical substances to which children are exposed during the fetal stage or early childhood will be identified. The JECS results will be translated to better risk assessment and management to provide healthy environment for next generations. The Japan Environment and Children\u2019s Study (JECS) is a nation-wide and government funded birth cohort study that started recruiting expecting mothers in January 2011. JECS is aimed to provide the foundation for policy making to safeguard the environment for the next generations.The Miami Declaration on Children\u2019s Environmental Health was adopted at the G8 Environment Ministers\u2019 Meeting held in Miami in 1997, in the midst of a growing concern regarding the effects that environmental pollution posed to children, and the acknowledgement of the vulnerability of children to harmful substances in the environment. The World Health Organization (WHO) published a report in 2006 in which approximately 40% of all children\u2019s death was attributed the environment . The UniIn Japan, the Advisory Board on Children\u2019s Environmental Health, established by the Ministry of the Environment (MOE), proposed a large-scale birth cohort study in order to evaluate the effects of environmental chemicals on children\u2019s health and development. In April 2008, the Working Group of the Epidemiological Research for Children\u2019s Environmental Health (later JECS Working Group) was organized and started systematic reviews on existing epidemiological findings regarding health impact of chemical exposures and the roles of potential confounders and effect modifiers, such as other environmental factors, genetic factors, socioeconomic status and lifestyle, in order to develop JECS study design and hypotheses. In March 2010, JECS Working Group published a draft conceptual plan for a large-scale birth cohort study covering all of Japan . The budThe ultimate goal of JECS is \u201cto identify environmental factors that affect children\u2019s health and development in order to help decision makers design better chemical risk management strategies\u201d.One of the major characteristics of JECS is that it is implemented as a national project funded directly by MOE, in contrast with most of the other epidemiological studies that are carried out in Japan by universities or research institutes using government research subsidies. The JECS is structured by three-tier centers. The National Center for JECS (program office), established in the National Institute for Environmental Studies (NIES), leads the JECS, while the National Center for Child Health and Development (NCCHD) supports the National Center as the Medical Support Center with its medical expertise. The National Center and Medical Support Center will cooperate together with 15 Regional Centers, located from the north, Hokkaido, to the south, Okinawa on epidemiologica studies of MOE, and Ethics Committees of all participating institutions, i.e., NIES, NCCHD, Hokkaido University, Sapporo Medical University, Asahikawa Medical College, Japanese Red Cross Hokkaido College of Nursing, Tohoku University, Fukushima Medical University, Chiba University, Yokohama City Unversity, University of Yamanashi, Shinshu University, Univresity of Toyama, Nagoya City University, Kyoto University, Doshisha University, Osaka University, Osaka Medical Center and Research Institute for Maternal and Child Health, Hyogo College of Medicine, Tottori Unversity, Kochi Univeristy, University of Occupational and Environmenatal University, Kyushu University, Kumamoto University, University of Miyazaki, and University of the Ryukyus. The JECS will be conducted in acoordance to the Helsinki Declaration and to the other nationally valid regulations.To ensure generalizability and ability to extrapolate the results of JECS to Japanese population, the 15 Regional Centers are selected to cover wide geographical areas. The study locations\u2019 urbanization and land development are diverse, from urban and suburban to rural areas as well as from agricultural and fishery to commercial and industrial uses.Regional Centers were selected in a competitive process in which universities and other research institutions were invited to submit proposals for covered areas and population, recruitment methods, organization structures, regional liaison, and the resources. Each Regional Center consists of one or more study areas. The population of the selected study areas is 130,000 to 600,000. Assuming birth rate of the study areas to be 1%, each Regional Center will see 1,300 to 6,000 annual births, 4,400 on average. JECS aims half of all the births in the area to be covered. Selected Regional Centers are required to recruit 3,000 to 9,000 pregnant women in three years, totaling to 100,000 participants in 15 Regional Centers is around 3% of Japanese newborns. Partners are also recruited but their participation is not mandatory.The eligibility criteria for participants (expecting mothers) are as follows: 1) They should reside in the study areas at the time of the recruitment, and are expected to reside continually in Japan for the foreseeable future, 2) expected delivery date should be between 1 August 2011 and mid-2014, and 3) they should be capable to participate in the study without difficulty, i.e., must be able to comprehend the Japanese language and complete the self-administered questionnaire. Those residing outside the study areas, even if they visit the cooperating health care providers within the study areas, are excluded from the study.We make contact with as many expecting mothers who reside in study areas as possible. The recruitment rate is targeted to be more than 50% of all eligible mothers. Either or both of the following two recruitment protocols are applied: 1) recruitment at the time of first prenatal examination at cooperating health care providers, i.e. obstetric facilities reproduction and pregnancy complications , 2) congenital anomalies , 3) neuropsychiatric disorders , 4) allergies and immune system deficiencies , 5) metabolism and endocrine system disorders . However, hundred thousand is not enough to analyze the association between environmental exposures and cancers. JECS collects cancer information in order to contribute future international pooled analysis, e.g. International Childhood Cancer Cohort Consortium (I4C) .From the JECS cohort, a sub-cohort with the size of 5,000 will be extracted. In that sub-cohort extended outcome measurements are planned, for instance, clinical analysis of blood samples from children; face to face interviews by medical staffs to evaluate neurological development; and medical examination.Environmental factors are manifold and complicated. In order to evaluate exposure to a wide range of environmental factors, the following four approaches are employed:1. QuestionnairesA part of each questionnaire is designated to collect information about chemical exposure, e.g. the use of organic solvents, kerosene, pesticides, disinfectants, heavy metals, antineoplastic drugs, narcotics, paints, hair dyes, and printer inks. Exposure to noise, vibration, high/low temperature, and dusts is also asked in the questionnaires.2. Chemical analysis of bio-specimensChemical substances or their metabolites are measured in peripheral blood, cord blood, breast milk, urine, and hair. Target compounds are shown in Table\u00a03. Environmental measurements2.5), will be measured during home visits. Noise levels and other physical parameters such as temperature and humidity will also be assessed.In the same sub-cohort as the one described above, indoor air pollutants, including volatile organic compounds (VOCs), aldehydes, nitrogen oxides, and fine particulate matters , nitrogen dioxide (NO2), and photochemical oxidants are monitored continuously. Twenty other hazardous air pollutants are also monitored at over 300 sites. Exposure to classical and hazardous air pollutants will be estimated from the monitoring station data using atmospheric simulation models.There are about 1,500 ambient air quality monitoring stations and about 500 roadside air quality monitoring stations across Japan, where levels of the five classical air pollutants, i.e., carbon monoxide (CO), suspended particulate matter (SPM), sulfur dioxide , lifestyle factors , and physical environment . Biochemical tests are performed on maternal, partners\u2019, and cord blood samples. Thyroid-stimulating hormone is analyzed for in dried blood spots from new born babies.All data collected from the participants are maintained by a data management system (DMS) developed by the National Center. Questionnaires are scanned and transformed to electronic data by optical character recognition (OCR) at the Regional Centers, then transferred to the DMS. Clinical chemistry data are electronically sent to the National Center and loaded on to the DMS. Personally identifiable information is stored separately from the data. All access to the DMS is recorded.Bio-specimens including blood, cord blood, urine, breast milk, and hair provided by the participating mothers, partners and their children are archived in three different storage facilities located in the National Center and elsewhere. Samples waiting for chemical analysis are stored in negative 80 degrees Celsius. Some aliquots of samples are reserved in liquid nitrogen tanks for much longer period for future analysis. A computer assisted repository system was developed in order to securely manage the bio-specimens for long-term. Chemical analysis will start after completion of the recruitment in early 2014.At this point, the JECS study is planned to continue until 2032, five years after all the participant children reach 13 years of age, allowing thorough data analysis. JECS may be extended beyond 2032 to further examine adolescence\u2019s health. The bio-specimen repository may be converted to a bio-bank upon completion of the JECS study to contribute further scientific research. All of these possibilities are written in the consent form.In addition to the JECS main study, adjunct studies are conducted by the National Center, Medical Support Center, Regional Centers, or any combination of them using their own funding. The adjunct studies may include procedures that are not adopted by the main study, e.g. collection and examination of placenta. Proposals for adjunct studies need to be approved by MOE, ensuring that they do not interfere with the main study.In recent years, there has been a growing concern regarding the vulnerability of children to harmful substances in the surrounding environment. Children are still undergoing development and the structure and function in each organ reach maturity at differing stages. Exposure to toxic chemicals during certain periods of development, namely critical windows, may lead to much severer consequences than the similar exposure in adulthood .The environment surrounding us has become quite different from what it used to be. The floors and walls of our houses are made of new materials. Even though floors are usually made from wood, their coatings which children touch may not be natural but artificial polymers. Our clothes have also changed from cotton and silk to polyester and acrylic fibers. Toys are no longer made from bamboo and wood but from plastics. The interior of residential houses, offices, and vehicles has also changed from traditional materials to industrial ones. Fire regulation now requires thermoplastics, thermosets, textiles and coatings be treated with flame retardants. Children eat processed foods, wrapped in plastics, and are living in the environment filled with novel chemicals. Even though those chemicals are thoroughly tested before they reach the market, we have little knowledge about their effects on children\u2019s health and development, especially when they exist as a mixture. The effect of those chemicals could be subtle. When one wants to examine not only such minute association of the chemicals with children\u2019s health but also the causality, a large scale prospective study is the only solution. JECS is an extremely ambitious project, run by the Japanese government, which is aimed to evaluate the impact of the environment in which our children live on their health and development. The fruit of JECS will therefore be with no doubt translated into better regulations and policies.Two birth-cohorts have been conducted in Japan, the Hokkaido Study of Environment and Children\u2019s Health and Tohoku Study of Child Development, provided a good basis for developing the JECS design. The Hokkaido Study began in 2002 and its population consists of 20,000 children . It starThe recruitment of JECS started in January 2011. Albeit Japan has suffered from severe damage caused by the mega-earth quake and tsunami, the number of participants increased steadily after March 2011. As of March 14, 2013, the number of enrollment reached 62,751 mothers and 28,982 partners. Mothers gave birth to 40,144 babies. Cord blood was collected form 38,008 new born babies. At the current recruitment rate, we expect to enroll 100,000 participants by early 2014 as planned.The budgets which had been spent to conduct JECS were 2.5, 4.6, and 6.1 billion yen in 2010, 2011, and 2012 Japanese fiscal years (starting April), respectively.Chemical analysis of bio-specimens and data analysis will start shortly after the completion of the recruitment in early 2014. The first publication that reports the association between environmental factors and some early stage outcomes such as pregnancy complications, congenital anomalies, and birth data will become available within the next few years.CO: Carbon monoxide; DMS: Data management system; I4C: International Childhood Cancer Cohort Consortium; IRB: Institutional review board; JECS: Japan Environment and Children\u2019s Study; MOE: The Ministry of the Environment; NCCHD: National Center for Child Health and Development; NIES: National Institute for Environmental Studies; NO2: Nitrogen dioxide; OCR: Optical character recognition; PM: Fine particulate matter; SO2: Sulfur dioxide; SPM: Suspended particulate matter; VOC: Volatile organic compound; WHO: World Health Organization.All the authors of this manuscript have no competing interests as defined by BioMed; we declare that we do not have any other interests that influence the results and discussion of this paper.The authors are justifiably credited with authorship, according to the authorship criteria. TK is primary investigator of this study. HiSat is ex-primary investigator. HN, KM, TK and HiSat conceived the study idea, designed the study. NT, ET, YK and MH carried out administrative procedure. RK, KM, FK and ZY supervised the study design and protocol development. YO, HiSai, HaS, TO, SY, MO and MT supervised the study from the point of medical and clinical aspect. YS and SN supervised the study from the point of exposure science. HN and AT contributed to develop the study protocol in the point of statistical view. TK, ET, SN, TM and AT wrote the draft and edited the manuscript. All authors participated in the discussion of the protocol development and revision of the manuscript. All authors critically revised the manuscript and approved the final version.Toshihiro Kawamoto is the director of National Center for Japan Environment and Children\u2019s Study (JECS), National Institute for Environmental Studies (NIES), and the professor of Department of Environmental Health, University of Occupational and Environmental Health.Hiroshi Nitta is the acting director of National Center for JECS, and the director of Center for Environmental Health Sciences, NIES.Katsuyuki Murata is the professor of Department of Environmental Health Sciences, Akita University Graduate School of Medicine.Eisaku Toda is the former director of Environment Risk Assessment Office, Environmental Health Department, Ministry of the Environment (MOE) and now the director of International Strategy Division, Global Environment Bureau, MOE, 1-2-2 Kasumigaseki, Chiyoda-ku, Tokyo 100-8975, Japan.Naoya Tsukamoto is the former director of Environment Risk Assessment Office, Environmental Health Department, MOE, and now the director of Industrial Waste Management Division, MOE, 1-2-2 Kasumigaseki, Chiyoda-ku, Tokyo 100-8975, Japan.Manabu Hasegawa is the former deputy director of Environment Risk Assessment Office, Environmental Health Department, MOE, and now a deputy director of Guidance of Medical Service Division, Health Policy Bureau, the Ministry of Health, Labour and Welfares, 1-2-2, Kasumigaseki, Chiyoda-ku, Tokyo 100-8916, Japan.Zentaro Yamagata is the professor of Department of Health Sciences, Interdisciplinary Graduate School of medicine and Engineering, University of Yamanashi.Fujio Kayama is the professor of Division of Environmental Toxicology, School of Medicine, Jichi Medical University.Reiko Kishi is the director of Hokkaido Unit Center for JECS, and a professor emeritus of Hokkaido University.Yukihiro Ohya is the acting director of Medical Support Center for JECS, National Center for Child Health and Development (NCCHD).Hirohisa Saito is the director of Medical Support Center for JECS and also the deputy director of National Research Institute for Child Health and Development, NCCHD.Haruhiko Sago, Makiko Okuyama and Susumu Yokoya are directors of NCCHD and also belong to Medical Support Center for JECS, NCCHD.Tsutomu Ogata was a director of NCCHD and is now the professor of Department of Pediatrics, Hamamatsu University School of Medicine, 1-20-1 Handayama. Higashi-ku, Hamamatsu-city, Shizuoka 431-3192, Japan.Yuji Koresawa is the former deputy director of National Center for JECS and is now the director of Office of Waste Disposal Management, MOE. 1-2-2, Kasumigaseki, Chiyoda-ku, Tokyo 100-8975, Japan.Yasuyuki Shibata is a researcher of National Center for JECS and senior scientist of NIES.Shoji Nakayama is a senior scientist of National Center for JECS and a chief of Center for Environmental Health Sciences, NIES.Takehiro Michikawa and Ayano Takeuchi are researchers of National Center for JECS and Center for Environmental Health Sciences, NIESHiroshi Satoh is the former director of National Center for JECS and now an acting director of Food Safely Commission, Cabinet Office, 5-2-20 Akasaka, Minato-ku, Tokyo 107-6122, Japan.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/14/25/prepub"} +{"text": "International Congress Health-Nutrition-Fitness-Wellbeing - SANABUNAINT 2012 took place in Falticeni, during 19-21 of October 2012. The event, which was organized under the patronage of the Romanian Patriarchy, the Romanian Academy, the Ministry of Public Health, the Ministry of Agriculture and Rural Development, the Ministry of Education, Research, Youth and Sport, has benefited from the presence of some important personalities. Lots of physicians took part in the event, many of whom being youngsters or even students.The second edition of the Apple Festival - \u201cThe Apples of Radaseni\u201d, and as an expression of the responsible preoccupation of the Falticeni Town Hall and Suceava Town Council in promoting the Public-Civic-Private Partnership, while considering the entire series of potential benefits , under the slogan \u201cPilot Station\u201d Falticeni and an assumed challenge: Constructing a Central Regional and Eastern-European Model of Thinking and Acting\u201d, was held for two days and has brought into light discussion themes such as the following: The Danube Delta - TD Education Research Innovation Development; Communication and Information Technology: Protecting, Promoting Health and Preventing Diseases \u2013 Re; Health-Nutrition-Fitness-Wellness - an actual challenge; etc. The event, which took place in the context of the 9th edition of the A section of round tables and a rich and useful explosion of medical books, belonging to the excellent and famous \u201cCarol Davila\u201d University Press, have completed the program. The works of the Congress were closed with the awards ceremony for the best papers and a magnificent show of \u201cCiprian Porumbescu\u201d folkloric group."} +{"text": "The published funding and competing interests statements were incorrect.The correct funding statement is: No current external funding exists for this study.The correct competing interests statement is: We have the following interests. CMFB, IJD, M-LB and ZVM-J were employees of WALTHAM Centre for pet nutrition during the course of this study and M-LB is currently at Mars Pet Care Europe. FED, EAK, ECT, JMB, TS were employees of the Forsyth Institute during the course of this study and LM is from the Veterinary Dental Surgery. FED has consulted for WALTHAM. FED CMFB, M-LB, and ZVM-J are inventors on patent WO2008137541, Dog Plaque Health, filed by WALTHAM. There are no other patents, products in development or other marketed products to declare. This does not alter our adherence to all the PLoS ONE policies on sharing data and materials, as detailed online in the guide for authors.The publisher apologizes for the error."} +{"text": "Lenke classification of idiopathic scoliosis (IS) is based on standing ap and lateral radiographs, combined with assessment of scoliosis correction on bending X-rays.The aim of this study was to verify the usefulness of bending films for Lenke classification of IS.The radiographs of 30 consecutive patients operated on because of IS were examined. Seven independent researchers assessed the X-rays, in 3 stages at one week intervals.Stage 1: Lenke type was determined on AP and lateral long film standing X-rays.Stage 2: Lenke type was established by use of AP and lateral standing X-rays, completed with supine traction films.Stage 3: Lenke type was indicated using AP and lateral standing, supine traction, and lateral bending films.The order of the radiographs, in each stage, was different and random. The results were determined by calculating the inter-observer and intra-observer agreement, and were quantified using two-rater and multi-rater kappa statistics.The inter-rater agreement in the first, and second, stage was moderate . The inter-rater agreement in the third stage was less (multi-rater kappa coefficient: 0.34 and percentage agreement: 0.47). The intra-observer agreement was the highest between the first and second stage for each of the researchers .The use of lateral bending X-rays in classifying the IS, according to the criteria of Lenke, reduced the intra-rater and the inter-rater agreement. Supine traction radiographs of the spine improved the agreement. This may suggest that the supine traction X-rays may help in classifying the IS."} +{"text": "There were errors in the Funding section. The correct funding information is as follows:MAS and co-authors are thankful to the Deanship of Scientific Research, King Saud University, Riyadh, Saudi Arabia, for supporting the work through the research group project number RGP-VPP-301. RC was supported by the Gebert R\u00fcf Foundation (grant number GRS-046/09). GS was supported by the French National Agency for Research(ANR), Association Contre les Syndromes C\u00e9r\u00e9belleux, France, The Verum Foundation, the European Union , The Fondation Roger de Spoelberch and the program \"Investissements d'avenir\" ANR-10-IAIHU-06 (to the Brain and Spine Institute). HA was supported by Association Fran\u00e7aise contre les Myopathies (AFM), France. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Protocol-based care of the tracheostomised patient is important, as adverse events confer a high rate of mortality. Little is known regarding the existence of formal evidence-based guidelines on tracheostomy care. The aim of this study was to perform a systematic review for evidence-based guidelines on adult tracheostomy care.A systematic search of PubMed, MEDLINE, guideline clearinghouses, centres of evidence-based practice, and professional societies' guidelines relating to care of adult patients with a tracheostomy was performed by two reviewers. In addition, a Google search of publicly available tracheostomy care guidelines was performed. Search terms: (tracheostom* OR tracheotom*) AND . Filters: English language, human, from 1 January 1990 to date, adult patients. Guideline appraisal criteria: the quality of guidelines retrieved was assessed using the Appraisal of Guidelines Research and Evaluation II (AGREE II) instrument [The search results are summarised in Table Five evidence-based guidelines on adult tracheostomy management were identified. This may represent a paucity of evidence on the subject, suggesting that further clinical trials on the topic are needed to contribute to the evidence base. This also highlights the need for international consensus on the topic, to reduce duplication of efforts, standardise practice, and improve outcomes."} +{"text": "During the production process of this article, the Academic Editor's attribution below the Citation was removed from the manuscript PDF. It should read:Editor: Toru Hosoda, Tokai University, Japan"} +{"text": "The authors would like to add additional funding. When Drs. Suhel Parvez and Julietta U. Frey were added to the author byline, their support from DFG and Alexander-von-Humboldt Foundation should have also been included in the financial disclosure for this article. The Funding section should read: \"This work was supported by NSFC and Scientific Innovation Projects from Fudan Univ. to TB and by the DFG , the CBBS / Land Saxony-Anhalt / EU (C1-TP4 to MRK), DIP grant, the DZNE (to MRK), and the Schram Foundation (to MRK). SP was supported by the Alexander-von-Humboldt Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "China is considered to be the main carbon producer in the world. The per-capita carbon emissions indicator is an important measure of the regional carbon emissions situation. This study used the LMDI factor decomposition model\u2013panel co-integration test two-step method to analyze the factors that affect per-capita carbon emissions. The main results are as follows. (1) During 1997, Eastern China, Central China, and Western China ranked first, second, and third in the per-capita carbon emissions, while in 2009 the pecking order changed to Eastern China, Western China, and Central China. (2) According to the LMDI decomposition results, the key driver boosting the per-capita carbon emissions in the three economic regions of China between 1997 and 2009 was economic development, and the energy efficiency was much greater than the energy structure after considering their effect on restraining increased per-capita carbon emissions. (3) Based on the decomposition, the factors that affected per-capita carbon emissions in the panel co-integration test showed that Central China had the best energy structure elasticity in its regional per-capita carbon emissions. Thus, Central China was ranked first for energy efficiency elasticity, while Western China was ranked first for economic development elasticity. In the 15th Conference of the Contracting Parties under the \u201cUnited Nations Framework Convention on Climate Change\u201d and the 5th Conference for the Parties under the \u201cKyoto Protocol\u201d held during December 2009 in Copenhagen, Denmark, the Chinese government solemnly promised that the carbon emissions per unit GDP would be decreased by 40\u201345% by 2020 from the 2005 levels. According to the \u201cKyoto Protocol\u201d agreed at the climate conference held in Copenhagen, there was a consensus about how to reduce greenhouse gas emissions and to keep the atmospheric temperature at a reasonable level. China is believed to be the highest carbon producer in the world, so it feels that it is imperative to transform its current status by implementing technological and systematic innovation, fundamentally abandoning its old-fashioned economic development pattern, and adopting an energy saving and low-carbon economic development path to improve its energy efficiency, thereby rationalizing people\u2019s lifestyles and consumption patterns.The \u201cTokyo Protocol\u201d and \u201cCopenhagen Annual Conference\u201d declared clear and concrete constraints on all nations regarding their carbon emission reduction obligations, but these issues are still under discussion as to how to evaluate a nation or district\u2019s carbon emissions, and which indicator should be utilized for scientific measurements. Many scholars and professionals have explored this issue in effective ways. Mielnik et al. proposed that the carbon dioxide emissions per energy unit could be used as the main evaluation criteria to address climate change and the economic development models of developing countries Decomposition analysis is often used in studies related to carbon emissions. For example, Fan et al. used the Adaptive Divisia Decomposition method (AWD) to divide the factors affecting China\u2019s carbon intensity during 1980\u20132003 and showed out that the primary energy intensity had a significant effect on the carbon intensity 2 emissions 2 emissions The LMDI approach is a type of index decomposition analysis (IDA), while another popular decomposition technique for energy and emissions is structural decomposition analysis (SDA). Three aggregation issues are inherent in SDA studies, namely sector aggregation, spatial aggregation and temporal aggregation The decomposition method is a relatively common method for the analysis of regional carbon emissions change. However, many studies merely decompose the carbon emissions into a number of factors without further exploration of the influential mechanisms and dynamic changes in these factors. Based on the results of factor decomposition for per-capita carbon emissions, we selected the Panel Co-integration Analysis method and conducted an in-depth exploration of the long-term and short-term dynamic changes, and the factors with strong effects on the per-capita carbon emissions.As mentioned above, the main indexes available for evaluating the regional carbon emissions situation include the total carbon emissions, per-capita carbon emissions, carbon emissions per unit GDP, and the carbon emissions performance based on the DEA model, while the per-capita carbon emissions indicator is very important for demonstrating the regional carbon emissions level. In summary, the LMDI decomposition approach proposed by Ang et al. is used more frequently for carbon emissions decomposition, besides the LMDI method has recently become popular in SDA studies related to energy and emissions. China\u2019s regional carbon emissions vary greatly, i.e., there was an eightfold difference in the per-capita carbon emissions of the highest and the lowest provinces in 2009, so we used the LMDI factor decomposition model\u2013panel co-integration test two-step method to study the main factors affecting the per-capita carbon emissions in Chinese provinces, and explore the influential mechanisms and dynamic changes in the per-capita carbon emissions, thereby facilitating a quantitative analysis of the emissions reduction strategy.This paper is organized as follows. Section 2 presents the results of the regional per-capita carbon emissions in Chinese provinces. Section 3 describes the LMDI approach used to decompose the per-capita carbon emissions and the decomposition results in China. Based on the factors decomposed in Section 3, the panel co-integration analysis of the factors that affect the regional per-capita carbon emissions is presented in Section 4. Finally, we conclude this study.This study analyzed the energy consumption of total coal, total oil, and natural gas, which were obtained from the China Energy Statistical Yearbook published by China\u2019s National Bureau of Statistics (CNBSa) According to the Kaya Identity, the regional carbon emissions can be decomposed as follows:Where, Eq. (3) shows that the regional per-capita carbon emissions are affected by four factors:Energy structure , the change in the regional per-capita carbon emissions can be decomposed into four factors:Energy structure factor: Carbon emission coefficient factor: Energy efficiency factor: Economic development factor: In this study, we used the LMDI decomposition method introduced by Ang et al. The decomposition factors are expressed as follows.Where, In Eq. (2), i represents the type of energy consumption, including total coal, total oil, and natural gas, which are all converted into standard coal. The data of the total coal, total oil, and natural gas for each region were taken from China Energy Statistical Yearbook The decomposition results show that the most powerful driver of the per-capita carbon emissions in Eastern China, Central China, Western China, and China between 1997 and 2009 was economic development. Undoubtedly, China\u2019s economy grew rapidly between 1997 and 2009, and the per-capita carbon emissions were closely associated with economic development. Thus, the rapid development of the economy led to vigorous growth in the per-capita carbon emissions, which was exemplified by the high average annual growth in China\u2019s per-capita carbon emissions (7.1%) between 1997 and 2009. The main effect of economic growth on the per-capita carbon emissions in Eastern China was similar in Central China, but greater than that in Western China. The inhibitory effect of energy efficiency on the increase in the per-capita carbon emissions was much greater than that of the energy structure. The inhibitory effect of energy efficiency improvement on the per-capita carbon emissions in Eastern China was similar to that in Central China, but larger than that in Western China. Furthermore, the inhibitory effect of the energy structure change on the increased per-capita carbon emissions was very weak and the shares of the energy structure factor in Eastern China, Central China, and Western China were all less than 1%. Indeed, the coal-oriented energy structure in the three economic regions remained unchanged. The proportion of coal was >70% and the only difference was a slight increase in the proportion of natural gas. The micro-adjustment of the energy structure had a limited effect on the change in the per-capita carbon emissions. For these provinces, the effect of economic growth on the per-capita carbon emissions was high in the top five provinces, i.e., Beijing, Tianjin, Shanghai, Jinlin, and Shanxi, which comprised three in Eastern China, one in Western China, and one in Central China. By contrast, the inhibitory effects of energy efficiency were highest in five other provinces: Beijing, Tianjin, Shanghai, Jinlin, and Shanxi, which comprised three in Eastern China, one in Central China, and one in Western China.The LMDI decomposition method was used to decompose the change in the per-capita carbon emissions between 1997 and 2009 into the energy structure factor, the energy efficiency factor, and the economic development factor. We also studied how these three factors affected the per-capita carbon emissions in Eastern China, Central China, and Western China using the panel co-integration test method. The basic idea of co-integration is to verify whether there is a long-term stable combined relationship between unstable variables. If this combination is also a stationary sequence, we can conclude that these variables have achieved co-integration relationship.The difficulty of the unit-root test for Panel data is that we consider the heterogeneity of the cross-section, but we also construct a higher potential statistic. The Panel data unit root test method is not unified and the more commonly used methods are the LLC test, Breitung test, IPS test, Fisher ADF, Fisher PP test, and Hadri test. Of these, the LLC test, Breitung test, and Hadri test are the same-root test methods, whereas the IPS test, Fisher ADF, and Fisher PP test are different-root test methods. Moreover, the LLC test, Breitung test, IPS test, Fisher ADF, and Fisher PP test assume that the unit root exists, whereas the Hadri test assumes no unit root.There are two ways of performing the co-integration test with Panel data: one based on the maximum likelihood ratio, and another based on the residual. It is well known that the Johansen Fisher test focuses on the maximum likelihood ratio, whereas the Pedroni test and Kao test prefer to use the residual of the E\u2013G two-step method. We used the Pedroni test.The Pedroni test is applied mainly to heterogeneous panels and it has seven co-integration statistics: four are interclass statistics and three are group statistics . Panel V, Panel Rho, Panel PP, Group Rho, Group PP Phillips, and Perron statistics use nonparametric tests, whereas Panel ADF and Group ADF use the ADF test. The null hypothesis of the seven statistical tests is no co-integration relationship whereas the alternative hypothesis of the interclass statistic requires a uniform co-integration coefficient for every cross-section unit and the group statistic allows variation.In this study, the energy structure factor (ES) is denoted by the coal share of the total energy consumption, energy efficiency factors are expressed as the energy consumption per unit GDP, and the economic development factors (RY) are represented by per-capita GDP. The data were derived from: (1) China Energy Statistical Yearbook published by China\u2019s National Bureau of Statistics (CNBSa) Using the six methods mentioned above for the panel data unit-root test, unit-root tests of the four variables are carried out for Eastern China, Central China, and Western China. Due to the article length limit, we do not list the unit-root test results for the three economic regions. The Hadri test results were not obvious and in many cases the level value, the first-order difference value, and the second-order difference value were all significant. Therefore, the Hadri test results were excluded. As stated previously by Harris and Tzavalis The unit-root test results showed that the four variables, i.e., the regional per-capita carbon emissions, energy structure, energy efficiency, and economic development, were not stable. Therefore, we tested whether the four variables had a co-integration relationship prior to the panel data regression, which would have caused spurious regressions. In this study, we tested whether there was a co-integration relationship between the four variables using the seven statistics included in the Pedroni test methods. The test results are shown in Like test results mentioned by Pedroni The co-integration test showed that there were co-integration relationships between the per-capita carbon emissions and the energy structure, energy efficiency, and economic development in each economic region. The long-term equilibrium model of the regional per-capita carbon emissions and its factors is as follows.Eastern China:Central China:Western China:Initially, for the energy structure elasticity involved with regional per-capita carbon emissions: Eastern China\u200a=\u200a0.615, Central China\u200a=\u200a0.767, and Western China\u200a=\u200a0.765. Thus, Central China ranked first, Western China second, and Eastern China third. After considering the energy efficiency elasticity, the results for Eastern China, Central China, and Western China were 0.867, 0.983, and 0.971 respectively, with the highest in Central China, the second in Western China, and the lowest in Eastern China. Finally, after considering the economic development elasticity, the results were 0.974 in Eastern China, 0.998 in Central China, and 1.017 in Western China, i.e., Western China, Central China, and Eastern China in descending order.The energy structure is characterized as the coal proportion in the overall energy consumption and coal is the highest of all the carbon emission coefficients of primary energy, so the energy structure elasticities of the per-capita carbon emissions were positive in Eastern China, Central China, and Western China. As a result, the energy structures in Eastern China, Central China, and Western China were 68.0%, 89.5%, and 76.4%, respectively, in 1997, but 59.6%, 83.8%, and 76.5%, in 2009. The energy structure and energy structure elasticity of the per-capita carbon emissions shared identical sequences in Eastern China, Central China, and Western China so the higher coal reserves and production in Central China and Western China led to a higher coal proportion of the energy consumption and a rapid increase in carbon emissions and per-capita carbon emissions.In conclusion, a higher energy intensity (energy consumption per unit GDP) was linked to higher per-capita carbon emissions, which agreed with our theoretical expectations. The energy intensity was 1.475, 2.044, and 2.439 in Eastern China, Central China, and Western China, respectively, as tons of standard coal per million yuan in 1997, which changed to be 1.149, 1.543, and 1.855 in 2009, i.e., decreases of 22.1%, 24.5%, 23.9%, respectively. The extent of the reduction kept pace with the energy efficiency elasticity of the per-capita carbon emissions in the three regions.The per-capita GDPs in Eastern China, Central China, and Western China were 9,284 yuan, 4,957 yuan, and 3,883 yuan, respectively, in 1997, which soared to 28,678 yuan, 14,639 yuan, and 13,038 yuan in 2009, with an annual growth rate of the per-capita GDP from 1997 to 2009 of 9.9%, 9.4%, and 10.6%. In general, a higher economic development level led to higher individual consumption levels and more carbon emissions were generated. Western China\u2019s per-capita GDP grew at the highest rate from 1997 to 2009. Due to the low level of economic development in Western China, its per-capita carbon emissions level ranked last among the three regions in 1997, whereas they exceeded those of Central China in 2009. Therefore, from 1997 to 2009, the economic development elasticity of the regional per-capita carbon emissions was highest in Western China but lowest in Eastern China, which experienced excessive per-capita carbon emissions accompanied by rapid economic development.Traditionally, an econometric model is specified based on a particular economic theory or the recognition of economic behaviors, which help to clarify the theoretical relationship between the model variables. However, the co-integration and error correction model determined the variables and the relationships between them by referring to specific relationships derived from the data with respect to economic variables. We used the Engle-Granger two-step approach to establish the panel error correction model and to examine the short-term dynamics of changes in China\u2019s per-capita carbon emissions. The model was as follows.Variations in the regional per-capita carbon emissions can be divided into long-term equilibrium and short-term fluctuations. The error correction term in the model reflects the intensity of adjusting the per-capita carbon emissions that deviate from the long-terms equilibrium. Eq. (11) was used to estimate the regional error correction models, as shown in The error correction model demonstrates that in the short-term the effects of the regional energy structure, energy efficiency (energy intensity), and economic development on the per-capita carbon emissions Western China, Central China, and Eastern China ranked first, second, and third, respectively. In addition, each region\u2019s error correction model (ECM) terms showed that the theoretical assumptions matched the reality. The ECM coefficients for Eastern China, Central China, and Western China were \u20130.0327, \u20130.0881, and \u20130.1700, respectively. Of these, Western China adjusted to the balanced state at the fastest rate, followed by Central China and Eastern China.The average annual growth of China\u2019s per-capita carbon emissions was 7.1% between 1997 and 2009. In 1997, Eastern China, Central China, and Western China ranked first, second, and third in the per-capita carbon emissions, while in 2009 the ranking changed to Eastern China, Western China, and Central China, where Western China exceeded Central China. The LMDI decomposition results showed that economic growth stimulated the increase of per-capita carbon emissions in Eastern China, Central China, and Western China between 1997 and 2009. It is notable that the effects of economic growth on the per-capita carbon emissions in Eastern China were similar in Central China, but greater in Western China. In addition, the inhibitory effects of energy efficiency on the increased per-capita carbon emissions were much higher than those of the energy structure. Furthermore, the inhibitory effect of improved energy efficiency on the per-capita carbon emissions in Eastern China were similar to that in Central China, but larger than that in Western China. The inhibitory effect of the energy structure\u2019s change on the increased per-capita carbon emissions was very weak, i.e., <1% in Eastern China, Central China, and Western China. The analysis of the factors that affected the regional per-capita carbon emissions using a co-integration test indicated that for the energy structure elasticity involved with the regional per-capita carbon emissions, Central China ranked first, followed by Western China and Eastern China. After considering the energy efficiency elasticity, Central China was highest, followed by Western China and Eastern China. Finally, when the economic development elasticity was considered, Western China was highest, followed by Central China and Eastern China.The LMDI decomposition results showed that the continued growth of GDP per capita was the dominant factor that led to the growth of per-capita carbon emissions, while the stimulating effect of economic development on the carbon emissions per-capita in Central China was greater than that in Eastern China, and that in Eastern China was also greater than that in Western China. As a developing country, the growth of the economic development and GDP per-capita is necessary to meet the national needs and development requirements, and energy consumption is the basic input that maintains the normal operation of the economic system. Thus, an increased pressure on the environment is inevitable. China\u2019s current stage of development means that the continued growth of China\u2019s carbon emissions and per-capita carbon emissions are unavoidable in the future for a long period of time. Furthermore, according to the current international division of labor, China produces many high energy-consuming products for developed countries, which makes it more difficult for China to access the peak phase of carbon emissions than developed countries. The results of the LMDI decomposition and co-integration analysis showed that economic development was the most important factor that affects the growth of per- capita carbon emissions. However, economic growth will not lead to a reduction in carbon emissions immediately, and it need not lead to the growth of total carbon emissions. Thus, it is possible to slow the growth in the per- capita carbon emissions through government-led emission reduction measures, which can control the carbon emissions growth during the economic development process.First, it is necessary to strengthen efforts to optimize the industrial structure and promote the development of low-carbon industries. Based on administrative and economic measures, governments can facilitate resource integration in the high energy-consuming heavy industry to limit the development of high-carbon industry. There should be an aim to develop tertiary industry because the energy intensity per GDP in the tertiary industry is less than one-quarter of that in the secondary industry. In 2012, China\u2019s tertiary industry accounted for 44.6% of the total, whereas it comprised two-thirds of the total in developed countries, so the development of China\u2019s tertiary industry still has great potential. There should be a focus on actively developing a modern service industry and high-tech industries, and using financial measures to support the development of the information industry, eco-industries, new energy development, and other green industries.Second, there is a need to fundamentally change the high input, high energy consumption, high pollution, and low efficiency growth pattern, and adopt an intensive growth mode instead of an extensive one. The LMDI decomposition results showed that one-quarter of the per-capita carbon emissions growth caused by economic development was offset by an increase in the energy efficiency, so energy savings and energy efficiency improvement are the most effective ways to reduce emissions. According to a study of China\u2019s Energy Development Strategy and Policy Research Group, the energy consumption per unit product in energy-intensive industries was 21% higher than that of the world\u2019s advanced level, which shows that there is still great potential for China to slow the growth of per-capita carbon emissions by continuing to improve the energy efficiency and reducing the carbon emissions per unit of GDP in the future. China should actively encourage the development and application of low carbon technologies, transform the steel, cement, and other high-carbon industries using high-tech approaches, promote industrial innovation and upgrading within the industry, and focus on the promotion and application of advanced emission reduction technologies. China can also strengthen international cooperation, introduce advanced energy-saving technologies such as clean coal technology from developed countries and promote them, as well as encouraging the use of the Clean Development Mechanism (CDM), carbon capture, and storage technology (CSS) in related units. Moreover, China can strengthen energy management to improve energy efficiency in many ways. For example, relevant laws and regulations could be established and improved to promote the development of a low-carbon economy, by setting enforced carbon intensity standards for various industries and by encouraging families to adopt low-carbon lifestyles by economic incentives.Third, there is a need to optimize the energy consumption structure. The decomposition results showed that the energy structure could limitedly slow the growth of per-capita carbon emissions, but this is because the coal-dominated energy consumption structure has not changed significantly in the last decade. China\u2019s coal-based energy consumption structure means that it is difficult to change rapidly. However, it is China\u2019s long-term aim to improve the energy structure by actively increasing oil and gas imports, developing new energy and renewable energy, and gradually reducing the weighting of coal in the overall energy consumption. China has abundant hydropower, wind, and solar energy, but the main factors that restrict renewable energy development at present are the cost and technology. Therefore, governments need to respond by supporting new energy through loans, taxes, and other measures. The growth of per-capita carbon emissions can be slowed by the establishment of a new \u201clow carbon\u201d or even \u201czero carbon\u201d energy system by developing and utilizing new energy and renewable energy sources.Finally, our results show that the current status and the trends in the per-capita carbon emissions in each region varied dramatically. Thus, efforts to narrow the gap in regional per-capita carbon emissions will help to lower their growth rate. To achieve the energy-saving objectives of China the circulation barriers should be eliminated, and low-carbon technologies could be shared within regions and resources to flow freely, thereby ensuring their optimal allocation.Appendix S1Supporting tables. Table S1, The LMDI-based decomposition results (1997\u20131998). Table S2, The LMDI-based decomposition results (1998\u20131999). Table S3, The LMDI-based decomposition results (1999\u20132000). Table S4, The LMDI-based decomposition results (2000\u20132001). Table S5, The LMDI-based decomposition results (2001\u20132002). Table S6, The LMDI-based decomposition results (2002\u20132003). Table S7, The LMDI-based decomposition results (2003\u20132004). Table S8, The LMDI-based decomposition results (2004\u20132005). Table S9, The LMDI-based decomposition results (2005\u20132006). Table S10, The LMDI-based decomposition results (2006\u20132007). Table S11, The LMDI-based decomposition results (2007\u20132008). Table S12, The LMDI-based decomposition results (2008\u20132009).(DOC)Click here for additional data file."} +{"text": "The PHEA-C16-iron oxide nanoparticles were synthesized by coprecipitation method. The core size of the PHEA-C16-iron oxide nanoparticles was about 5 to 7 nm, and the overall size of the nanoparticles was around 20, 60, and 150 nm in aqueous solution. The size of the nanoparticles was controlled by the amount of C16. The 3.0-T MRI signal intensity of a rabbit lymph node was effectively reduced after intravenous administration of PHEA-C16-iron oxide with the size of 20 nm. The in vitro and in vivo toxicity tests revealed the high biocompatibility of PHEA-C16-iron oxide nanoparticles. Therefore, PHEA-C16-iron oxide nanoparticles with 20-nm size can be potentially useful as T2-weighted MR imaging contrast agents for the detection of lymph nodes.The purpose of this study was to synthesize biocompatible poly(2-hydroxyethyl aspartamide)\u2013C Accurate diagnosis of lymph node metastasis in various cancer patients is very important as it is one of the most important factors for the choice of preoperative chemoradiotherapy, surgical treatment, and patient prognosis . Recentl3O4) nanoparticles for MRI applications. PHEA is a synthetic polymer having a protein-like structure, obtained by the reaction of ethanolamine with polysuccinimide (PSI), which is prepared by thermal polycondensation of d,l-aspartic acid. PHEA has good biopharmaceutical properties as drug carrier such as high water solubility, multi-functionality, absence of toxicity, antigenicity, immunogenicity, and low cost of production pyrene (+S) or ethylmethanesulfonate (\u2212S), positive responses were observed. Therefore, it was concluded that PHEA-C16-iron oxide did not induce chromosomal aberrations in the CHL cells which were used in this study.There were no statistically significant increases in the frequencies of aberrant metaphases with structural or numerical aberrations in the PHEA-C16-iron oxide nanoparticles with sizes of 20, 70, and 150 nm, controlled by the amount of C16 as a hydrophobic side chain. In vitro and in vivo experiments were performed with 20-nm particles; USPIO was used because the purpose of this study was to develop a novel MRI contrast agent for the detection of lymph nodes. The PHEA-C16-iron oxide as compared with Resovist\u00ae, a clinically approved MRI contrast agent, showed slightly better imaging contrast in lower concentration of iron from the in vitro phantom test. The PHEA-C16-iron oxide could also effectively detect bone marrow and lymph node of the rabbit by MRI. A single-dose intravenous toxicity and gene toxicity study of the PHEA-C16-iron oxide was commissioned by the preclinical research center of ChemOn, Inc. The results of LD50 from single-dose toxicity test were calculated 1,500 mg/kg for males and 1,300 mg/kg for females. PHEA-C16-iron oxide was confirmed biocompatible and nontoxic because genetic toxicity tests show negative results. These results indicate the great potential applications of PHEA-C16-iron oxide for the detection of lymph nodes by MRI.In this study, we have prepared biocompatible PHEA-CThe results suggest that the PHEA-C16-iron oxide has slightly better imaging contrast effect than the clinically approved contrast agent Resovist\u00ae. The PHEA-C16-iron oxide has excellent imaging contrast in the liver, blood vessel, and even in the lymph node. With its biocompatible and nontoxic characteristics, the PHEA-C16-iron oxide is a promising candidate for a new contrast agent.We certify that there is no conflict of interest with any financial organization regarding the material discussed in the manuscript.in vitro MRI experiments. BKK carried out the in vivo MRI experiments. BCS and SHC took part in the synthesis of the nanoparticle compounds. SHY carried out the NMR characterization analysis. YWC assisted with the toxicity experiments. CWC oversaw the study and provided insightful guidance. All authors read and approved the final manuscript.As the main author, DCH conceived of the study and participated in its design and overall experiments. HYL drafted the manuscript and participated in the design of the study. YWS carried out the DCH gained his MS degree in 2005 at the Department of Food Science, Kongju National University, Gongju, Republic of Korea. HYL attained his PhD degree in 2006 at the Department of Advanced Organic Materials Engineering, Chonbuk National University, Jeonju, Republic of Korea. YWS received her BS diploma in 2012 at the Department of Chemistry, Chungnam National University, Daejeon, Republic of Korea. SHY gained his PhD degree in 1987 at the Department of Physics and Chemistry, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea. YWC attained his PhD degree in 2007 at the College of Pharmacy, Kangwon National University, Chuncheon, Republic of Korea. BKK gained his PhD degree in 2002 at the Department of Radiology, Chung-Ang University, Seoul, Republic of Korea. BCS achieved his PhD degree in 1998 at the Department of Physics and Chemistry, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea. CWC gained his PhD degree in 1996 at the College of Pharmacy, Chonnam National University, Kwangju, Republic of Korea. SHC received his PhD degree in 2000 at the Department of Physics and Chemistry, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea."} +{"text": "AbstractEmesopsisinfenestra Tatarnic, Wall & Cassis, 2011 (Heteroptera: Reduviidae) is reported from New Zealand for the first time, based on a single specimen collected alive in the wild in Auckland in June 2013. The species was previously known only from Australia (Queensland) and the Loyalty Islands . Emesopsisinfenestra Tatarnic, Wall & Cassis, 2011 was originally described from Australia and the Loyalty Islands (2 specimens). Nothing else has been published about it. As far as I am aware, it has never before been collected from New Zealand.Tatarnic, Wall & Cassis, 2011Type status:Other material. Occurrence: recordedBy: Stephen E. Thorpe; individualCount: 1; Location: country: New Zealand; stateProvince: Auckland; verbatimLocality: Tamaki Campus (East), suburb of Saint Johns; verbatimLatitude: 36.88685S; verbatimLongitude: 174.85260E; Event: eventDate: 2013-06-10; Record Level: institutionCode: Auckland Museum (AMNZ)Emesopsisinfenestra from the original description (http://demo.nzor.org.nz/names/4330e783-5a90-4552-8427-e0bf56a027c3), the New Zealand fauna of Reduviidae comprises species of the genera Empicoris, Ploiaria and Stenolemus. I therefore recommend that Emesopsisinfenestra be added to the New Zealand Organisms Register (NZOR) as present in the wild. Single specimen records can be problematic, but this is largely because most of them are processed long after they were collected, as part of routine curation, and so there is always the possibility of mislabelling with other samples. In this case, however, I personally processed and photographed the specimen within hours of having captured it, so I am confident that there is no possibility whatsoever of mislabelling or contamination. The biostatus (indigenous or exotic) of the species in New Zealand is uncertain. On the one hand, the specimen was found in a highly anthropogenic habitat, which argues for an exotic origin. On the other hand, since the sample size is so low and the species widespread , it is impossible to infer very much at all, and so the species might well be indigenous to New Zealand.On 10 June 2013, I collected a single specimen of an emesine reduviid amongst long grass in a weedy overgrown wasteland area within the Tamaki Campus (East) of the University of Auckland. It is easily identified as cription , and NikPloiariaantipoda, but fully macropterous, and clearly different to any of the known species in New Zealand. I found it crawling up a spider web covered tree trunk at night.As an aside, there is another unrecorded and as yet unidentified emesine species present in New Zealand. I collected a single specimen in Auckland Domain about 8 years ago and deposited in the New Zealand Arthropod Collection (NZAC). As I no longer have access to NZAC, I cannot check the details, but it was a large species, similar to the native"} +{"text": "Although impaired health-related quality of life (HRQOL) has been reported in patients with sarcoidosis, there is currently no sarcoidosis-specific questionnaire in Japan. The 29-item Sarcoidosis Health Questionnaire (SHQ), originally developed in the United States, is the only sarcoidosis-specific HRQOL questionnaire currently available. The primary aim of this study was to develop and validate a Japanese version of the SHQ.The SHQ was translated into Japanese following the forward-backward procedure. The reliability and validity of the Japanese version of the SHQ were examined. One hundred twenty-two Japanese patients with biopsy-proven sarcoidosis were evaluated by the SHQ, the Medical Outcomes Study 36-item short form (SF-36), the St. George's Respiratory Questionnaire (SGRQ), chest radiography, an electrocardiogram, laboratory blood tests, pulmonary function tests, an echocardiogram, and assessments of dyspnea and depressive symptoms. The SHQ was found to have acceptable levels of internal consistency . SHQ scores correlated significantly with scores on the SF-36 and SGRQ. The domain or total scores on the SHQ also significantly correlated with serum levels of the soluble interleukin-2 receptor, the percentage of the predicted forced vital capacity, pulmonary arterial systolic pressure, dyspnea, and depressive symptoms. Also, the SHQ scores of patients who had one or two organ systems affected by sarcoidosis were significantly different from those of patients who had three or more organ systems involvement.The Japanese version of the SHQ can be used to assess the HRQOL of patients with sarcoidosis. Sarcoidosis is a chronic and multisystem granulomatous disease of unknown etiology that can involve almost any organ system . The assThe Sarcoidosis Health Questionnaire (SHQ), developed by Cox and coworkers in the United States (US), is the only sarcoidosis-specific HRQOL questionnaire . They reWe consecutively enrolled 138 Japanese patients with biopsy-proven sarcoidosis who presented to the Kyoto Central Clinic between June and December 2009. Exclusion criteria included age <18 years, presence of an active neoplasm, and cognitive and/or reading impairment that prevented completion of the questionnaires.All patients underwent chest radiography, an electrocardiogram, laboratory blood tests, pulmonary function tests (PFTs), and an echocardiogram. The current extent of organ involvement was assessed by the A Case-Control Epidemiologic Study of Sarcoidosis (ACCESS) Organ Involvement Index . Chest rHRQOL was assessed by the Japanese versions of the SF-36 ,16 and tData are presented as mean \u00b1 standard deviation. Cronbach's coefficient \u03b1 was used to assess the reliability of the questionnaire. Spearman's rank correlation test was used to examine the relationships between SHQ scores and the other HRQOL measures (SF-36 and SGRQ scores) and the clinical variables measured. The Mann-Whitney U test was used to compare HRQOL questionnaire scores between patients who differed in the number of sarcoidosis-affected organ systems (organ system involvement). Values of p < 0.05 were considered statistically significant.Of the 138 enrolled patients, 16 were excluded because they either did not complete the questionnaires sufficiently for assessment or they had no active organ involvement of sarcoidosis at that time. Table Cronbach's coefficient \u03b1 values of the DF, PF, and EF domains of the SHQ were 0.85, 0.68, and 0.77, respectively, and the \u03b1 value for the total SHQ score was 0.91. The SHQ total score, as well as the scores for all three domains, were essentially normally distributed . They also weakly to strongly correlated with the three component scores and the total score of the SGRQ . The DF, EF, and total SHQ scores weakly correlated with sIL2-R levels , while the DF, PF, and total SHQ scores correlated with the percentage of the predicted forced vital capacity , but only the PF score correlated with the PASP . All three domain scores and the total SHQ score moderately correlated with scores on the MRC dyspnea scale and the CES-D scale .The relationships between SHQ scores and the other HRQOL measures and the clinical variables measured are presented in Table Patients who had involvement of one or two organ system(s) had significantly lower scores than patients who had three or more organ systems involved in the DF domain (p = 0.01), the EF domain p = 0.03), the total SHQ score (p = 0.04), and in three of the eight subscales of the SF-36 (p < 0.01 to p = 0.04) scores on other HRQOL questionnaires and 2) clinical variables in Japanese patients with sarcoidosis.The reliability of the translated SHQ was assessed for internal consistency. The Cronbach's \u03b1 values for all three domains and the total SHQ were above the 0.60 required to support construct validity , indicats| = 0.27 to 0.75). These findings indicate that while the translated SHQ evaluated the HRQOL of Japanese patients with sarcoidosis, it did not necessarily measure the same aspects as the other HRQOL questionnaires. The SHQ scores were weakly but significantly associated with a serum biomarker (sIL-2R), pulmonary function, and the echocardiographic index. These correlations indicate that the SHQ can evaluate the effects of multiple pathophysiological processes in sarcoidosis that involve several organ systems. Furthermore, the SHQ scores more highly correlated with patient-reported outcomes such as dyspnea (MRC) and depressive symptoms (CES-D) than with physiological measurements. These findings are consistent with our previous findings in other diseases [The correlations between SHQ scores and SF-36 and SGRQ scores were weakly to strongly significant (|rdiseases ,22,23. IThe validity of the SHQ was also shown by comparing the scores between patients who differed in the extent of organ system involvement. The SHQ scores were significantly different between patients with one or two organ system involvement and those with three or more. Furthermore, compared with the SF-36 and the SGRQ, the SHQ was better able to distinguish the HRQOL between these two groups.The clinical manifestations of sarcoidosis may differ between different ethnic groups. Compared to Western countries, sarcoidosis in Japan is less severe, but has a higher likelihood of ocular and cardiac involvement ,24,25. TWe have demonstrated the reliability and validity of a Japanese version of the SHQ. The SHQ can evaluate the HRQOL of Japanese patients with sarcoidosis, and when added to routine radiological, serological and physiological evaluations of the disease, can provide valuable additional information. As the SHQ is a disease-specific questionnaire, it can evaluate what the generic SF-36 and respiratory-specific SGRQ do not evaluate.1: forced expiratory volume in one second; FVC: forced vital capacity; HRQOL: health-related quality of life; LVEF: left ventricular ejection fraction; SF-36: Medical Outcomes Study 36-item short form; MRC: Medical Research Council; %DLCO: the percentage of the predicted diffusion capacity for carbon monoxide; %FEV1: the percentage of the predicted forced expiratory volume in one second; %FVC: the percentage of the predicted forced vital capacity; PF: Physical Functioning; PASP: pulmonary arterial systolic pressure; PFT: pulmonary function tests; sACE: serum angiotensin-coverting enzyme; SHQ: Sarcoidosis Health Questionnaire; sIL-2R: soluble interleukin-2 receptor; SGRQ: St. George's Respiratory Questionnaire.ACCESS: A Case-Control Epidemiologic Study of Sarcoidosis; ACE: angiotensin-coverting enzyme; BHL: bilateral hilar lymphadenopathy; CES-D: Center for Epidemiologic Study-Depression Scale; DF: Daily Functioning; DLCO: diffusion capacity for carbon monoxide; EF: Emotional Functioning; FEVThe authors declare that they have no competing interests.Full responsibility for the integrity of the data and the accuracy of the data analysis: THConception and design: THAnalysis and interpretation of the data: KT, TH, TODrafting of the article: KTCritical revision of the article for important intellectual content: TH, SN, TO, TK,YI, KW, KA, KC, MM, TIFinal approval of the article: SN, KC, MM, TICollection and assembly of the data: KT, TH, TK, YI, SNAll authors read and approved the final manuscript."} +{"text": "The crucial procedure includes surface treatment of polystyrene core particles by cationic polyelectrolyte polyethyleneimine, in situ formation of Ag nanoparticles, and immobilization of the Ag nanoparticles onto the surface of the polystyrene colloids via functional group NH from the polyethyleneimine. The experimental parameters, such as the reaction temperature, the reaction time, and the silver precursors were optimized for improvement of dispersion and Ag coat coverage of the core-shell-shaped nanostructures. Ultimately, the optimum parameters were obtained through a series of experiments, and well-dispersed, uniformly coated PS/Ag core-shell-shaped nanostructures were successfully fabricated. The formation mechanism of the PS/Ag core-shell-shaped nanostructures was also explained.In this paper, based on the previous steps, a facile Recently, considerable effort has been devoted to controllable fabrication of nanostructured materials with tunable functional properties. One example is the fabrication of core-shell-shaped nanostructures. Such nanocomposite structures have many applications in different technological fields like bio-sensor, chem-sensor, electronics, catalysis, drug delivery, diagnostics, antibacterial agent, etc. -7. Espec2 are used to activate the core surface, but they remain as an impurity in the final product. The layer-by-layer self-assembly is too complicated and time-consuming. In the sonochemical method, the high-intensity ultrasound and removal of oxygen are necessary; otherwise, impurity like Ag2O can be observed.The properties of the Ag-coated CSSNs are dependent on metal coverage. Thus, control of metal coverage is important to the application of this kind of core-shell-shaped nanostructure . Until nin situ reduction method with optimized fabrication parameters was developed for the fabrication of polystyrene/Ag (PS/Ag) CSSNs. In the first step, monodispersed PS colloids as core particles were prepared by emulsion polymerization and then modified with polyethyleneimine (PEI) several times. Subsequently, Ag seeds were formed in situ and immobilized on the surface of PS colloids by adding AgNO3 or silver ammonia solution into the PEI-modified PS colloids. Finally, sodium citrate was added in the dispersion to increase thickness of the Ag shell. In this work, key experimental parameters were optimized to improve the dispersion and Ag coverage of PS/Ag CSSNs. Finally, well-dispersed and uniformly coated PS/Ag CSSNs were obtained. The thickness and coverage of the Ag shells can be easily controlled by changing the temperature, time, and silver precursors by which the properties of PS/Ag CSSNs can be tailored.In this paper, based on the previous investigations , an adva2S2O3), sodium citrate (C6H5O7Na3\u00b72H2O), ethanol (C5H6OH), and sodium lauryl sulfate (CH3(CH2)SO4Na) were purchased from Shantou Dahao Fine Chemical Co., Ltd. ; silver nitrate (AgNO3), ammonia, PEI (MW600000-1000000) were purchased from Shanghai Chemicals Co. Ltd. . Water used during the experiments was distilled twice.Styrene, potassium pyrosulphate operating at 20 kV. The chemical compositions of particles were examined by X-ray energy-dispersive spectra (EDS). Zetasizer Nano Essentials was used to measure the size and potential of particles.Surface morphology of the original PS particles to serve as cores for the coating fabricated by emulsion polymerization is shown in Figure Since these PS particles are covalently bonded whereas the nature of bonds in Ag is metallic, it is difficult to bond Ag atoms directly onto the PS surface. Therefore, surface modification with functional groups is essential for the purpose. Thus, cationic polyelectrolyte PEI was used to attach functional group of NH on the PS surface as shown in Figure \u22121 is due to the presence of hydroxyl group on the PS spheres, while the other absorption bands at 3,028, 2,925, 1,608, 1,497, 1,454, 760, and 700 cm\u22121 are due to the asymmetric, symmetric, and deformed vibrations of the -CH2 group of the benzene ring. It is obvious that the broad absorption peak at 3,412 cm\u22121 is typical of the N-H bond stretching vibrations, and peaks at 1,601, 1,498, and 1,450 cm\u22121 stem from stretching vibrations of the C-C bonds in the benzene ring [Figure ene ring . The IR + ions with the sodium citrate. The products were washed by deionized water where the excess PEI was removed, and thus, the isolated core-shell structure can be obtained. We can see that the obtained PS/Ag CSSNs are well-dispersed, uniformly coated with a good coverage of Ag where the coated metallic Ag is not oxidized as confirmed in Figure Figure 3 was added into the PEI-modified PS colloids, stirred or seeded at different temperatures for 1 h, and subsequently reduced by sodium citrate for another 30 min at 80\u00b0C as shown in Figure Our study shows that different fabrication parameters have obvious influence on the structure of the PS/Ag CSSNs. For this reason, we studied systematically crucial influences of the key parameters on the structure of the PS/Ag CSSNs. First, the influence of temperature was investigated, and the results are shown in Figure Figure + ions in the solution which could transport to the Ag crystal nuclei as the time increased. However, the system seemed to reach the state of saturation with the PEI after the modification in a certain time later. As a result, the best time for the Ag seeding for the advanced fabrication of the PS/Ag CSSNs should be longer than 1 h.The seeding time is another important factor in the fabrication, which was found to have a particular effect on the Ag coverage of the PS/Ag CSSNs. We can see this obviously from Figure + in solution in many cases can be critically important for the rational control over surface and interfacial modifications of the molecular absorption process. The commonly used polyelectrolyte binders are very sensitive to their local ionic environment [3 were used widely in the latest papers. The detailed mechanism for the difference is under a further exploration.The environment of the Agironment . Our exp3 at 100\u00b0C at least for 1 h followed by the reduction via sodium citrate at 80\u00b0C for 30 min leads to the well-dispersed, uniformly coated PS/Ag CSSNs. During the in situ reduction, the Ag seeds immobilized on the surface of the PEI-modified PS colloids through the linkage of functional group N-H. The dispersion and the Ag shell coverage of the PS/Ag CSSNs can be easily controlled by changing the temperatures of seeding and reduction, the seeding time, and the kind of silver ions. This optimized process is crucial to fabrication of PS/Ag CSSNs and their applications.The experimental results showed that the stirring of the PEI-modified PS colloids with AgNOThe authors declare that they have no competing interests.CZ performed the experiment, analyzed the data, and wrote the paper. XZ designed the experiment, analyzed the data, and wrote the paper. HL, IK, and MI helped write and revise the paper. LW and JB helped design the experiment. XC helped draft the manuscript. All authors read and approved the final manuscript.Scientific Journal of Physical Science, associate editor of the International Journal of Molecular Engineering and is on the editorial board of several journals such as the Chinese Science Bulletin, etc. He is also an active referee for several top international journals such as Applied Physics Letters, Journal of Physical Chemistry, Crystal Engineering Communications, Journal of Materials Chemistry, and Chinese Physics Letters. His current research interests are focused on nanoinstabilities of low-dimensional nanostructures under external excitations, energetic beam nanoprocessing, controllable fabrication and growth of low dimensional nanostructures, controllable assembling and construction of large scale arrays of zero-dimensional nanostructures, organic and inorganic hybrid at nanoscale, functionalization of nanostructure, etc. He has authored and co-authored over 100 publications, filed 8 patents, chaired, co-chaired, or served as committee or advisory board member at over 20 international or national conferences, and presented over 60 invited lectures and talks at universities, research institutes, and major international conferences worldwide.HL is an engineer in the College of Materials at Xiamen University with her research interest focused on advanced energy materials.IK is a Ph.D. student in condensed matter physics in the China-Australia Joint Laboratory for Functional Nanomaterials at Xiamen University with his research interest focused on processing of nanomaterials. MI is a Ph.D. candidate in condensed matter physics in the China-Australia Joint Laboratory for Functional Nanomaterials at Xiamen University with his research interest focused on fabrication of nanomaterials. LW is a professor and research director in the ARC Centre of Excellence for Functional Nanomaterials at University of Queensland and a visiting professor in the China-Australia Joint Laboratory for Functional Nanomaterials at Xiamen University. His current research interest is concentrated on advanced functional nanomaterials. JB is an associate professor in the State Key Laboratory of Polymer Materials Science and Engineering at Sichuan University and a visiting associate professor in the China-Australia Joint Laboratory for Functional Nanomaterials at Xiamen University with his research interest in polymer materials. XC is a professor in the College of Materials at Xiamen University and a deputy director of the China-Australia Joint Laboratory for Functional Nanomaterials at Xiamen University with her research interest focused on advanced functional nanomaterials.CZ is an M.S. candidate in materials science and engineering in the China-Australia Joint Laboratory for Functional Nanomaterials at Xiamen University with her research interest focused on fabrication of nanomaterials. XZ is a Ph.D. in electronic materials engineering at the Australian National University and one of the earliest scientists who initialized nanoresearch in China. He is presently the director of the China-Australia Joint Laboratory for Functional Nanomaterials, an adjunct professor at The University of Queensland in Australia, and a full-time professor in the School of Physics and Mechanical and Electrical Engineering at Xiamen University in China, as well as the chief scientist for the AMAC International Inc., USA. He is the editor-in-chief of the"} +{"text": "Human Rhinoviruses (HRVs) have high genetic diversity and three species have been described: HRV-A, HRV-B, and the recently recognized HRV-C, which has been rapidly identified worldwide.In the present study, we report the frequency and diversity of Human Rhinovirus (HRV) strains circulating in Panama from children hospitalized with respiratory infections.HRVs of species A, B and C have been identified with a predominance of HRV-A and HRV-C over HRV-B, and marked genetic diversity within each species. Human rhinoviruses (HRVs) are the most common causative agents of upper respiratory tract infections, but are also associated with more severe diseases such as pneumonia or acute wheezing related to bronchiolitis and acute asthma in children -4. HRV iEnterovirus, family Picornaviridae.The authors declare that they have no competing interests.JA and JP conceived of the study, and participated in its design and coordination. DF, LA, MC, MC, CC and JC carried out the PCR and sequencing studies and drafted the manuscript. AD, JA and DF participated in the phylogenetic analysis and JA wrote the paper. All authors read and approved the final manuscript."} +{"text": "Although significant advances have been made in implementation science, comparatively less attention has been paid to broader scale-up and spread of effective health programs at the regional, national, or international level. To address this gap in research, practice and policy attention, representatives from key stakeholder groups launched an initiative to identify gaps and stimulate additional interest and activity in scale-up and spread of effective health programs. We describe the background and motivation for this initiative and the content, process, and outcomes of two main phases comprising the core of the initiative: a state-of-the-art conference to develop recommendations for advancing scale-up and spread and a follow-up activity to operationalize and prioritize the recommendations. The conference was held in Washington, D.C. during July 2010 and attended by 100 representatives from research, practice, policy, public health, healthcare, and international health communities; the follow-up activity was conducted remotely the following year.Conference attendees identified and prioritized five recommendations (and corresponding sub-recommendations) for advancing scale-up and spread in health: increase awareness, facilitate information exchange, develop new methods, apply new approaches for evaluation, and expand capacity. In the follow-up activity, \u2018develop new methods\u2019 was rated as most important recommendation; expanding capacity was rated as least important, although differences were relatively minor.Based on the results of these efforts, we discuss priority activities that are needed to advance research, practice and policy to accelerate the scale-up and spread of effective health programs. Recognizing the need for better methods to accelerate adoption of effective health practices and programs, researchers and funding agencies have expanded work in implementation science and the related disciplines of improvement science and heali.e., scale-up or spread) in hundreds or thousands of institutions or communities is limited. Numerous practice-based efforts to scale-up and spread evidence-based health programs have been documented , but this work often does not employ theory-based, rigorous scientific approaches for studying scale-up processes, and thus offers limited evidence and guidance for improving future scale-up efforts. The terms \u2018scale-up\u2019 and \u2018spread\u2019 lack accepted, universal definitions[To date, however, most implementation studies have been conducted in relatively small- to moderately-sized samples of institutions, delivery systems, agencies, and/or communities-9. Althoinitions,10-16; winitions. Others initions,19. Deveinitions,20.Although limited, research interest in scale-up and spread is increasing. Much of this work has been conceptual or descriptive: developing frameworks and models for scale-up and spread,17,21-26nd Annual National Institutes of Health Conference on the Science of Dissemination and Implementation (2009). The dinner attendees proposed a state-of-the-art/agenda-setting conference involving approximately 100 U.S. and international representatives from research, practice, and policy in healthcare and public health, which we conducted during July 2010. The conference generated specific recommendations for actions needed to facilitate enhanced interest and activity in scale-up. A follow-up activity was conducted during Fall 2011 to prioritize and operationalize the recommendations. This article describes the methods and findings from the conference and the follow-up prioritization activity.Responding to these challenges and the need for expanded research, practice, and policy to ensure that effective programs achieve impact on health at the population level, we launched a multi-stakeholder initiative to increase awareness and to identify specific actions needed to expand scale-up activity in health. The initiative was envisioned and launched during an informal, 30-person working dinner meeting held in junction with a panel session at the 2The Conference to Advance the Science and Practice of Scale-up and Spread of Effective Health Programs in Healthcare and Public Health (hereafter noted as \u2018the conference\u2019) was held in Washington, DC from July 6-8, 2010. The conference was organized by representatives from the Institute for Healthcare Improvement , the University of Alabama at Birmingham School of Public Health (Norton), and the US Department of Veterans Affairs Quality Enhancement Research Initiative (Mittman). Approximately 100 individuals were invited to attend the conference, reflecting a purposeful mix from the research, practitioner, policymaker, public health, healthcare, U.S., and international communities . The 126 individuals included attendees from the conference as well as approximately 25 other individuals identified by colleague referrals. Of the 126 eligible individuals, 49 (39%) completed the survey.The five major conference recommendations, their associated sub-recommendation \u2018action items,\u2019 and participant importance ratings are listed in TableStakeholders noted the need for a rich portfolio of follow-up activities to maintain engagement and enthusiasm and to stimulate interest and involvement by additional stakeholders. Although the multi-stakeholder conference itself was intended to stimulate greater interest and activity, the conference attendees and survey respondents recognized the need for sustained follow-up efforts. Highly-rated suggestions included targeted, intensive outreach and education of the need for focused attention on scale-up and spread, as well as financial incentives and support.e.g., email groups, conference calls, meetings) for tracking and sharing information regarding ongoing policy, practice, and research in scale-up. This system would facilitate increased communication and collaboration among key stakeholders, more rapid learning and progress, and greater efficiency in resource utilization. Additional recommendations included the development of practical summaries of existing knowledge and research results to facilitate their use and benefits.Stakeholders noted that a considerable amount of activity in scale-up research, practice, and policy is not widely known, and thus fails to achieve its full benefit. They recommended creating a database and related mechanisms compared to methods to scale-up complex interventions across larger, interdependent systems . Stakeholders also emphasized the need to better understand conditions under which sufficient demand for scale-up of innovations will arise to complement or replace the supply-oriented approaches typically employed in small-scale, local implementation initiatives. Specific actions recommended to facilitate this knowledge production include the development of new funding opportunities for practice-based, practice-oriented research, the identification and expansion of existing programs supported by such research, and research efforts to develop taxonomies of scale-up strategies and factors influencing their effectiveness.Stakeholders recognized the need to further test and refine existing methods for effectively scaling-up health practices and programs, including a better understanding of when, where, and how particular methods are more or less effective. For example, research is needed to assess the effectiveness of methods to spread simple practices in specific settings , and thus might be biased and emphasize actions of interest to researchers and stakeholders in developed countries.Limitations of the recommendations and corresponding sub-recommendations generated from the conference and follow-up activity should be noted. The recommendations resulted from open discussion and brainstorming, and are neither comprehensive nor exhaustive; we welcome additional suggestions and \u2018action items\u2019 for advancing the field. The recommendations and their priority ratings reflect the composition of conference and survey participants and practice-based activities . The recommendations and sub-recommendations described herein are meant to specify actions to advance research, practice, and policy in scale-up and spread in health, and galvanize interest and effort in this area that are commensurate to its need.In sum, the initiative described herein brought together expert researchers, practitioners, and policymakers to identify and prioritize next steps for advancing the field in a way that sought to minimize isolated projects conducted within individual silos, capitalize on the depth of knowledge and expertise available, and stimulate future activity. As an extension of the initiative, the authors are launching several follow-up activities to pursue these recommendations, including development of working groups charged with tackling the identified priority recommendations and sub-recommendations, delineated by a focus on research-based activities (Implementation Science. CJM is now at the Centers for Medicare & Medicaid Services; he has reviewed the material herein for factual accuracy but has not made substantive contribution since summarizing the meeting and its findings prior to leaving the IHI. MWS reports no conflicts of interest. BSM is Editor-in-Chief Emeritus of Implementation Science.WEN is on the Editorial Board of All authors contributed to the development, organization, and execution of the state-of-the-art conference. CJM wrote the first draft of the summary of the conference. BSM, WEN, and MWS were responsible for conducting the follow-up online rating study. WEN drafted the first version of the full meeting report; BSM and MWS revised the manuscript. CJM reviewed the revised manuscript for factual accuracy. All authors read and approved the final manuscript."} +{"text": "Acute leukemia with coexisting Gilbert's syndrome treated by allogeneic hematopoietic stem cell transplantation is rarely reported. Here we described a case whose transaminase levels were almost normal, although transient hyperbilirubinemia repeatedly happened during chemotherapy. To the editor:We have seen a 52-year-old man with AML-FAB M2a subtype, who had no history of viral hepatitis. He had history of mild indirect hyperbilirubinemia with normal transaminase levels after he took paracetamols in the past two years, and the same phenomenon occurred to his siblings, children and nephews. He received three cycles of chemotherapy containing daunorubicin, idarubicin, pirarubicin, cytarabine, and obtained CR in the first cycle. His bilirubin level was normal before chemotherapy; however, mild non-hemolytic indirect hyperbilirubinemia happened to him during each cycle of chemotherapy Figure . NeitherAfter 3 cycles of chemotherapy, the patient received transplantation from his HLA-identical sibling sister who was also diagnosed to have GS. The conditioning regimen included fludarabine and busulfan. Hematopoietic engraftment was observed on day +11. In the absence of GVHD, the levels of transaminase and bilirubin were almost normal within 100 days post-transplantation (Figure GS is a common condition; its prevalence has been described in 3-10% of the general population . SeveralSome reports suggest that GS might be a risk factor of cancer ,6. GS wiTo our knowledge, there is little description of the onset of hyperbilirubinemia in GS patients who received HSCT. Ruiz-Arguelles GJ, et al. reportedGS: Gilbert's syndrome; allo-HSCT: Allogeneic hematopoietic stem cell transplantation; AML: acute myeloid leukemia; CR: complete remission; CT: Computed tomography; UGT1A1: uridine 5'-diphosphoglucose glucuronosyl transferase; HLA: human leukocyte antigen; GVHD: graft-versus-host disease; G-CSF: granulocyte-colony-stimulating factor; PCP: pneumocystis carinii pneumonia.The authors declare that they have no competing interests.All authors were involved in the provision of clinical care of the patient and the collection of data and the review of the manuscript. GPY: supplied the acquisition of data, analysis and interpretation of data, drafting of manuscript; QFL: provided the conception and design of the paper, revised it critically for important intellectual content, and final approval of the version to be submitted.QFL, MD, Ph.D, Department of Hematology, Nanfang Hospital, Southern Medical University; Member of the Asian-Pacific Society of Hematology; Member of Blood Branch of the Chinese Medical Association; Member of the Chinese hematopoietic stem cell transplant group; Standing Committee member of Chinese Anti-Cancer Association of Professional Committee of Hematology Branch; editor of more than 10 journals."} +{"text": "The Levels of Emotional Awareness Scale (LEAS) was developed to assess five levels of emotional awareness: bodily sensations, action tendencies, single emotions, blends of emotion, and combinations of blends. It is a paper and pencil performance questionnaire that presents 20 emotion-evoking scenes. We developed a Japanese version of the LEAS (LEAS-J), and its reliability and validity were examined.The LEAS-J level was independently assessed by two researchers who scored each response according to the LEAS scoring manual. High inter-rater reliability and internal consistency were obtained for the LEAS-J. Measures were socioeconomic status, LEAS-J, Toronto Alexithymia Scale-20 (TAS-20), Interpersonal Reactivity Index (IRI), and NEO Five-Factor Inventory (NEO-FFI). TAS-20, IRI and NEO-FFI were the measures used to explore the construct validity of LEAS-J, as it was predicted that higher scores on the LEAS-J would be related to fewer alexithymic features, greater empathetic ability, and a greater sense of cooperation with others. Questionnaires were completed by 344 university students.The criterion-referenced validity was determined: a significant negative relationship was found with the externally-oriented thinking scores of TAS-20, and positive relationships were found with fantasy, perspective taking, and empathic concern on IRI and with extraversion, openness to experience, and agreeableness on NEO-FFI.Consistent with our expectations, the findings provide evidence that the LEAS-J has good reliability and validity. In addition, women had significantly higher scores than men on LEAS-J, showing that the gender difference identified in the original LEAS was cross-culturally consistent. The conscious awareness of one's own emotions is a prerequisite for emotional intelligence, including the conscious regulation of one's own emotional states and expressive behaviors ,2. HigheThus, a reliable and valid method is needed for assessment of awareness of emotions, particularly one that is applicable in a variety of cultural contexts. Lane & Schwartz proposedA number of methods, such as self-report questionnaires and structured interviews, have been developed for assessing difficulty in identifying and describing specific emotional states -9. Self-In applying the LEAS to other countries, such as Japan, with cultures that may be quite different than that of the United States, it is important to take into account that the scenarios, which describe situations focusing on interpersonal relationships, need to be modified to fit the appropriate cultural context. Further, because emotional processes are influenced by cultural differences in attention and perception, cultural regulation of emotions should be considered when examining the level of emotional awareness . With thThe LEAS-J was given to 380 Japanese students, 344 (90.5%) of whom completed the questionnaires. They ranged in age from 18-38 y.o. [mean age (SD) = 20.13 (1.64)], and were recruited from two coeducational and one female-only university, all located in urban, middle class areas . The greater ratio of females to males resulted, in part, from the fact that women were recruited at all three sites, whereas men were recruited from only two.The aim of the study was explained in a classroom setting. The students were told that any student who registered to participate would be paid a small fee for completing the questionnaires at home and for sending them back by mail. Because it was too time-consuming in the classroom setting to complete the test battery that included the numerous short essays required by LEAS, the participants were requested to complete them at home. They were informed in writing that their privacy would be completely protected and that no participant would be identified when the results of the study were published. This study was approved by the local ethics committee of the National Center of Neurology and Psychiatry.The participants reported the occupations of their parents, and the 123 reported occupations were divided into a hierarchy of high, middle, and low class, based on the occupational prestige scores for Japan proposed by Tsuduki . For exaThe LEAS is a performance-based measure of the levels of emotional awareness ,6 Table. Some ofThe Japanese translation of the 20-item Toronto Alexithymia scale (TAS-20) developed by Komaki, et al. was usedThe Interpersonal Reactivity Index (IRI) is a self-report questionnaire that measures the empathetic ability of the participants ,16. EachThe NEO-Five Factor Inventory (NEO-FFI), an abridged version of the NEO Personality Inventory, is one of the standard measures of the big five factor model of personality structure . The fivLEAS-J was scored by two authors , working independently, who classified the emotion-related words attributed to Self and Other for each scenario. Before scoring the protocols, training was undertaken by both scorers who discussed the results of pilot testing until accurate scores were achieved, based on the LEAS scoring manual .Classification was according to the LEAS scoring manual . We usedTo extract emotion-related words from the data of the LEAS-J, we used the KHCoder, a Japanese corpus analysis software that higSPSS ver.11.0 was used for all statistical analysis. Statistical significance was set at p < 0.05, two-tailed.Intra-class correlation coefficients [ICC ] were calculated to determine the inter-rater reliability of each item and subscale of the LEAS-J Table . The resCronbach's \u03b1 coefficients were calculated for the Self, Other, and Total responses to the 20 scenarios as independently scored by two raters . Table T-tests were used for comparisons between males and females. The LEAS-J scores of females were higher than those of males on all subscales Table . DiffereA one factor analysis of variance (ANOVA) was completed for the comparisons based on socioeconomic status. The effect of the parents' position in the job hierarchy on the subjects' LEAS-J score was investigated. No significant effect on the LEAS-J score was observed for the father's or mother's occupation Table .Correlation coefficients were calculated for the mean score of LEAS-J , TAS-20 , IRI , and NEO-FFI scores (five factors). Table With regard to IRI, the LEAS-J Self, Other, and Total scores were significantly, positively correlated with FS , PT , and EC . No significant correlations with PD were observed.For the NEO-FFI scales, LEAS-J Self, Other, and Total scores were significantly correlated with E , O , and A . No significant correlations were found with N and C.We compared the LEAS-J scores with the LEAS scores obtained from a group of Americans students of the same age. The LEAS-J scores were lower than LEAS scores on all subscales . Although the relationship with TAS-20 showed rather poor correlation, except for EOT, the results for NEO-FFI and IRI indicate that the LEAS-J has good concurrent validity. The current findings confirm that the Japanese written descriptions of each scenario are essentially equivalent to the original, which indicates that the LEAS-J would be useful for assessing the level of emotional awareness of Japanese subjects.The LEAS-J was shown to have good internal consistency. The finding for \u03b1 coefficients was very similar to the findings of a previous study by Lane, et al. , suggestIn the age cohort comparison of LEAS-J and LEAS scores, however, the mean scores of almost all of the LEAS-J scenes were significantly lower than those of the LEAS. This could be due to a number of factors. The original LEAS was given to students in a classroom setting, while the LEAS-J subjects were requested to complete the scenarios at home. The difference in the collection method might have had some influence on the difference. Another factor may be the Japanese style of expressing feelings, which has been reported to be much different from that of people in Europe and the U.S. . JapanesIn addition to scoring by the glossary of the original LEAS as stated above, even after the same subjects' words and descriptions were re-scored assessing the individual level of emotional awareness as a whole at the conceptual level, the mean scores of all scenes on the LEAS-J remained significantly lower than the level of the mean LEAS scores. Nisbett & Masuda comparedConstruct and concurrent validity were obtained by examining the relationship between LEAS-J scores and TAS-20, IRI, and NEO-FFI scores and are as follows.The LEAS-J Self score showed a significant, negative correlation only with the EOT scores of TAS-20. This finding is similar to the finding that the LEAS subscales were negatively correlated with only the EOT of patients with somatoform disorders ,31 and tThe present study is the first report of the relationships between LEAS-J and IRI and NEO-FFI. The significant correlations of all aspects of LEAS-J with fantasy (FS), perspective taking (PT), and empathic concern (EC) in IRI suggest that the ability to take into account the feelings of oneself and others with sensitivity underpins the imaginative function that places others in the position of oneself (FS), an ability associated with understanding the inner emotional state (PT) of others and of consideration for others (EC).The LEAS-J scales were significantly associated with the NEO-FFI scores for Extraversion (E), Openness-to-Experience (O), and Agreeableness (A). Extraversion includes sociability, liveliness, and the general experience of positive affect. Openness-to-Experience includes aesthetic sensitivity, intellectual curiosity, need for variety, non-dogmatic attitudes, high curiosity and interest in the internal and external world. These positive associations with E and O are consistent with their negative associations with TAS-20 ,35. In aThe scores of women were higher than those of men for all of the LEAS-J subscales. This is consistent with the report by Barrett, et al. , and it In this study, the LEAS-J scale scores were not significantly different by socioeconomic status. However, Lane, et al. ,34 foundFurther work is necessary in the following areas: LEAS-J should be given to subjects with a wider range of ages to examine the possible influence of age as an indicator of emotional development on LEAS-J scores; to determine if LEAS-J adequately detects impairments of emotional awareness in clinical samples, such as patients with alexithymia; to develop an additional scoring system that is optimally suited to the Japanese language and culture to supplement the current glossary that is derived from LEAS protocols completed by subjects in the United States and is based on a direct translation from English; and finally, the reliability and validity of LEAS-J will need to be examined carefully by a concurrent structured interview, such as the SIBIQ .A Japanese version of LEAS, LEAS-J, was developed and its reliability and validity were examined. Although a Japanese specific glossary or a modified scoring method will be helpful in the future to capture culture-specific aspects of emotional life in Japan, the current results showed high inter-rater reliability and internal consistency. Construct and concurrent validity were supported by the relationships with TAS-20, IRI, and NEO-FFI. Women had higher scores than men on LEAS-J, supporting the cross-cultural consistency of this gender difference.The authors declare that they have no competing interests.TI collected and analyzed the data, performed the statistical analysis and drafted the manuscript. GK participated in the study design, analysis and interpretation of data and revision of the manuscript. RD made substantive contributions to the study design and interpretation of the results. YM, HN participated in the design of the study and collected the data. HA analyzed the data and YT provided advice on the data analysis. CS participated in the translation of the Japanese version and checked the revised scenarios. MM participated in the acquisition of data and made substantive contributions to the study design. All authors have read and approved the final manuscript"} +{"text": "There is information missing from the Funding section. The complete Funding statement is: \"This work was partly supported by grants CGL-2004-03300, CGL-2007-65515, and a FPI pre-doctoral fellowship for MH (BES-2005-6925) from the Spanish Ministry of Science and Innovation; grant IT542-10 from the Basque Government for Research Groups in the Basque University System, and UFI 11/09 from the University of the Basque Country UPV/EHU. No additional external funding was received for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There were errors in the Funding section. The correct funding information is as follows: Support for this project was provided by the National Institute of Allergy and Infectious Disease, National Institutes of Health, Department of Health and Human Services , the World Bank, and the Food and Agriculture Organization of the United Nations. This work was made possible through support provided by the Office of Health, Infectious Disease and Nutrition, Bureau for Global Health, U.S. Agency for International Development and Wildlife Conservation Society, under the terms of Leader Award No.LAG-A-00-99-00047-00, Cooperative Agreement: GHS-A-00-06-00005. The opinions expressed herein are those of the author(s) and do not necessarily reflect the views of the U.S. Agency for International Development or Wildlife Conservation Society. DOJ was supported by generous funding from the Dunemere private foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The last decade has seen widespread retreat from user fees with the intention to reduce financial constraints to users in accessing health care and in particular improving access to reproductive, maternal and newborn health services. This has had important benefits in reducing financial barriers to access in a number of settings. If the policies work as intended, service utilization rates increase. However this increases workloads for health staff and at the same time, the loss of user fee revenues can imply that health workers lose bonuses or allowances, or that it becomes more difficult to ensure uninterrupted supplies of health care inputs.This research aimed to assess how policies reducing demand-side barriers to access to health care have affected service delivery with a particular focus on human resources for health.We undertook case studies in five countries . In each we reviewed financing and HRH policies, considered the impact financing policy change had made on health service utilization rates, analysed the distribution of health staff and their actual and potential workloads, and compared remuneration terms in the public sectors.We question a number of common assumptions about the financing and human resource inter-relationships. The impact of fee removal on utilization levels is mostly not sustained or supported by all the evidence. Shortages of human resources for health at the national level are not universal; maldistribution within countries is the greater problem. Low salaries are not universal; most of the countries pay health workers well by national benchmarks.The interconnectedness between user fee policy and HRH situations proves difficult to assess. Many policies have been changing over the relevant period, some clearly and others possibly in response to problems identified associated with financing policy change. Other relevant variables have also changed.However, as is now well-recognised in the user fee literature, co-ordination of health financing and human resource policies is essential. This appears less well recognised in the human resources literature. This coordination involves considering user charges, resource availability at health facility level, health worker pay, terms and conditions, and recruitment in tandem. All these policies need to be effectively monitored in their processes as well as outcomes, but sufficient data are not collected for this purpose. Universal health coverage (UHC) has been labelled, 'the most powerful unifying single concept that public health has to offer\u2019, by Margaret Chan, Director of the World Health Organization.a. The primary initial focus of UHC policies has been on the extension of financial protection against health care costs through the provision of insurance and the removal of user fees at the point of use.UHC has become an international policy. In 2007, universal access to reproductive health was included among the Millennium Development Goals , which were unanimously agreed by all UN member states as part of the Millennium Declaration;b[http://www.icpdtaskforce.org/about/mission-vision.html];cService Availability and Readiness Assessment: see [REFD];d[http://www.who.int/healthinfo/statistics/indbirthswithskilledhealthpersonnel/en/] (accessed 15 March 2013); ethe narrow definition includes only midwives, doctors and clinical officers. The broad definition includes midwives, doctors, clinical offers and nurses weighted for the percentage of obstetric workload in the total facility workload;fThe World Health Report, 2005 (p91) suggests that for a district with a birth rate of 30/1000, one full-time-equivalent doctor is required for 3600 births. Gabrysch et al. (2011) translates this into 1200 births per doctor on the basis of three doctors required to provide 24-hour cover. The WHO Making Pregnancy Safer model specifies 1000 births per doctor, and we apply this lower number, which also seems to allow for professional development days, leave and sick leave;gPWT 7.0 Alan Heston, Robert Summers and Bettina Aten, Penn World Table Version 7.0, Center for International Comparisons of Production, Income and Prices at the University of Pennsylvania, May 2011. [http://pwt.econ.upenn.edu/php_site/pwt70/pwt70_form.php] (accessed 15 December 2011);h[http://data.worldbank.org/indicator/NY.GNP.PCAP.PP.CD] (accessed 21 December 2011);i[http://data.worldbank.org/indicator/NY.GDP.PCAP.CD] (accessed 15 December 2011);jdetailed income distribution data are not available but these estimates are based on the extreme assumption that those earning the given ratio of salaries to average GDP/GNI per capita capture virtually the whole GNI/GDP. More realistic assumptions rank the salary earner more highly still, relative to the rest of the population.ANC: Antenatal care; ANM: Auxiliary nurse midwive; CHO: Community health officer; CI: Concentration index; DFID: Department of International Development (UK); FBO: Faith-based organization; FHCP: Free Health Care Policy; GDP: Gross domestic product; GNI: Gross national income; HDI: Human development index; HMIS: Health management information system; HR: Human resource; HRH: Human resources for health; ILO: International labour organisation; IPCD: International Conference on Population and Development; LCM: Living Conditions Measurement Survey; MCH: Maternal child health; MDG: Millennium Development Goals; MNCH: Maternal and newborn child health; MNH: Maternal and newborn health; NGO: Non governmental organization; NHIS: National Health Insurance Scheme; OPD: Out patients; PNC: Postnatal care; RMNH: Reproductive maternal newborn health; SARA: Service availability and readiness assessment; SBA: Skilled birth attendant; STD: Sexually transmitted disease; TBA: Traditional birth attendant; UHC: Universal health coverage; WHO: World Health Organization.The authors declare that they have no competing interests.BM led the team, coordinated the research and was the lead author in drafting the paper. SW led the Ghana, Nepal and Zimbabwe case studies, contributed to cross-country analysis and commented on drafts of the paper. TE led the Zambia case study, led the cross-country analysis and commented on drafts of the paper. SF led the literature review work and commented on drafts of the paper. DN led the Sierra Leone case study and commented on drafts of the paper. TM contributed to the Nepal case study and commented on drafts of the paper. YC undertook fieldwork in Zimbabwe and commented on drafts of the paper. All authors read and approved the final manuscript.BM is Professor and Director of the Institute for International Health and Development, Queen Margaret University, Edinburgh. Within the ReBuild research consortium on health systems development, she is a research director, and leading research on the effects of health-financing policy change on the household economies of the poor. She is a health economist specializing in health policy and health systems research in low- and middle-income countries. SW is Reader at the Institute for International Health and Development, Queen Margaret University, Edinburgh. Within the ReBUILD research consortium on health systems development she is leading research into health worker incentives in the post-conflict period. She is a health economist specializing in health financing in low- and middle-income countries, with a focus on reducing financial barriers and improving the delivery of services through effective purchasing and provider payment mechanisms. TE is Head of the Nuffield Centre for International Health and Development at the University of Leeds and Professor of International Health Systems Research. He is a health economist with experience of teaching, research and consultancy in more than 20 low- and middle-income countries. His work has focused mainly on health financing, particularly costing of services, informal health-care markets, demand-side mechanisms and health-system equity. SF is Senior Lecturer at the Institute for International Health and Development, Queen Margaret University, Edinburgh. Within the ReBUILD research consortium on health systems development she is leading research into aid effectiveness in fragile settings. She is a social scientist focusing on governance and health systems development in fragile states and other unstable settings. DN is Senior Lecturer in Economics at Aberdeen University. He is an international health economist, working principally in West Africa and South Asia, with a particular interest in the productivity costs of maternal ill-health. Within the ReBUILD research consortium, he is supporting research on innovative contracting arrangements for health-care provision in Cambodia and Sierra Leone. TM is a Senior Lecturer in Human Resource Management at the Liverpool School of Tropical Medicine, UK. He is the Co-research Director for the DFID-funded ReBUILD research consortium on health systems in post-conflict States and leads on the human resources theme. He is also Principal Investigator for PERFORM which is investigating ways of improving health workforce performance at district level in three African countries. He has carried out consultancies in HRH in many countries and is currently advising the Ministry of Health and Population in Nepal. YC is a social scientist and currently Senior Research fellow with the Biomedical Research and Training Institute Centre for International Health and Policy (BRTI-CIHP). He is leading ReBUILD research in Zimbabwe on health-worker incentives and rural deployment and posting of human resources for health. He has extensive experience in poverty assessment and was involved in the formulation and implementation of the Second Poverty Assessment Study Survey (Pass II) in Zimbabwe in 2003 and in the production of the provincial and national reports."} +{"text": "There were errors in the Funding statement. The first sentence of the Funding Statement should read: \"This work was supported by the Biotechnology and Biological Sciences Research Council via the Centre for Integrative Mammalian Physiology and Pharmacology and the National Centre for Replacement, Refinement and Reduction of Animals in Research.\"In addition, in the Author Contributions statement the name of the second author was incorrectly abbreviated and the fourth author was incorrectly omitted from the list of those authors who conceived and designed the experiment. The Author Contributions statement should read: \"Conceived and designed the experiments: FA, SW, SS. Performed the experiments: FA. Analyzed the data: FA, KS. Contributed reagents/materials/analysis tools: CET, SW, SS. Wrote the paper: FA, CET, KS, SW, SS.\""} +{"text": "Within the context of a mission of the regional Health/Environment departments, CEDDES produced a second training CD-ROM in november 2010, about hazardous health care waste : infectious, chemical, toxic, radioactive waste.To respond to needs for training the different categories of staff in health care , education establishments, research centres in human medicine and veterinary, and professionals of the environment, in order to set up an optimal health care waste management.Stages :- Selection of the Steering Committee - period of the mission : 18 months- Document retrieval- WritingProduction of a CD-rom of 400 screens divided in 5 chapters: HCW, infectious HCW, chemical, toxic HCW, radioactive HCW, general waste; illustrated with 300 photos; glossary of 250 words; repertoire of 240 acronyms; proposals for evaluation quiz; selection of websites.The CD-rom can be used in developed countries as well as developing ones.A suitable HCWM is a complex system with regulatory, organizational, structural, budgetary components; with sanitary, environmental, economic, legal implications.Key elements: - Hand hygiene, control of health care associated infections, in the context of the new world stakes of patient safety; - Management of sharps and AES control.The CD-rom is a teaching aid to set up a long-lasting change of behaviour towards a common culture of safety. It got the Prize 2012 of the STHSS.None declared."} +{"text": "The Beta-Blockers in Heart Failure Collaborative Group (BB-HF) was formed to obtain and analyze individual patient data from the major randomized controlled trials of beta-blockers in heart failure. Even though beta-blockers are an established treatment for heart failure, uptake is still sub-optimal. Further, the balance of efficacy and safety remains uncertain for common groups including older persons, women, those with impaired renal function and diabetes. Our aim is to provide clinicians with a thorough and definitive evidence-based assessment of these agents. We have identified 11 large randomized trials of beta-blockers versus placebo in heart failure and plan to meta-analyze the data on an individual patient level. In total, these trials have enrolled 18,630 patients. Uniquely, the BB-HF group has secured access to the individual data for all of these trials, with the participation of key investigators and pharmaceutical companies.Our principal objectives include deriving an overall estimate of efficacy for all-cause mortality and cardiovascular hospitalization. Importantly, we propose a statistically-robust sub-group assessment according to age, gender, diabetes and other key factors; analyses which are only achievable using an individual patient data meta-analysis. Further, we aim to provide an assessment of economic benefit and develop a risk model for the prognosis of patients with chronic heart failure.This paper outlines inclusion criteria, search strategies, outcome measures and planned statistical analyses.http://clinicaltrials.gov/ct2/show/NCT00832442Clinical trial registration information: Heart failure (HF) is a major public health problem with both incidence and prevalence rising rapidly along with associated healthcare costs, estimated to be $39.2 billion in the United States and \u00a3625 million per year in the UK ,2. HF acHowever, survey data have confirmed that the uptake of beta-blockers in HF patients is still sub-optimal. Although the percentage of eligible patients prescribed beta-blockers increased between the first and second Euro Heart Failure surveys, a substantial number of patients remain untreated or receive sub-maximal therapy ,6. ParadAlthough a number of sub-group and meta-analyses based on published data have been conducted ,9, theseThe BB-HF group is a collaborative, multinational effort to combine individual data from the major randomized controlled trials (RCTs) investigating the use of beta-blockers in chronic HF. The group consists of the leading investigators of these trials and international experts, with the support of the four pharmaceutical companies that have marketed beta-blockers in HF . A full list of collaborators is presented in Appendix A. Two meetings of the collaborative group in November 2008 and August 2010 were used to define our objectives, establish inclusion criteria and develop the primary and secondary objectives. A standardized data request form was generated to obtain IPD from each eligible trial have been received by the coordinating center, the Clinical Trials and Evaluation Unit, Royal Brompton & Harefield NHS Trust/Imperial College London.The aims and objectives of the BB-HF individual patient data meta-analysis include:a) Provide a definitive estimate of the overall treatment effect of beta-blockers in HF on key outcomes including all-cause mortality and hospitalization.b) Analyze the influence of important pre-randomization patient characteristics on the clinical effects of beta-blockers, including age, diabetes, gender, ejection fraction, renal function, atrial fibrillation and the etiology of HF.c) Pool the adverse event and discontinuation data to assess the safety of beta-blockers in HF patients, particularly the rate of bradycardia and hypotension.d) Explore the relationship between the effects of beta blockers and key baseline measurements including heart rate, blood pressure and weight.e) Perform an exploratory analysis to assess important post-randomization variables, such as change in heart rate, achieved heart rate, change in blood pressure, achieved dose and the prescription of other concomitant therapies, such as ACE inhibitors, angiotensin receptor blockers (ARB), aldosterone antagonists, diuretics and digoxin.f) Describe in detail the effect on hospitalization data including recurrent hospitalization, total number of cardiovascular/HF-related hospitalizations and duration of stay according to treatment allocation.g) Develop a risk model for patients with HF using key baseline characteristics to allow clinicians to accurately assess prognosis in individual patients.h) Examine the potential economic impact of treatment under a variety of circumstances and different subgroups.i) Improve statistical methodology for IPD meta-analysis.A number of inclusion criteria were adopted to make the project methodologically sound and technically feasible. All RCTs of beta-blockers versus placebo explicitly reporting mortality as a primary or combination outcome will be included but not head-to-head comparisons with another active agent. Trials must include patients with documented symptomatic HF and only unconfounded trials will be accepted (in which one treatment group differed from another only by the beta-blocker therapy of interest). Finally, RCTs would only be included if they recruited more than 300 patients in total and planned follow-up of six months or greater. By concentrating on the larger trials , the project remains practical while the amount of data missed by not including the smaller trials is minimized.To ensure a complete assessment of the evidence, published or unpublished RCTs were identified through computer aided searches , scrutiny of reference lists of trials, trials registries, meeting abstracts, review articles and discussion with members of the collaborative group and with the pharmaceutical manufacturers.The search identified 11 trials that met all inclusion criteria ; the Australia/New Zealand Heart Failure Study (ANZ) , the BetThe primary outcome for BB-HF will be all-cause mortality, including deaths recorded after publication of the trial, where these data are available. The major secondary outcome will be the composite of all-cause mortality and cardiovascular hospitalizations. Secondary mortality outcomes include death due to acute myocardial infarction (MI), stroke, sudden cardiac death and HF-related death. Other secondary outcomes are non-fatal MI, all-cause hospitalization, cardiovascular hospitalization, HF-related hospitalization and the number and duration of hospital admissions. Drug safety outcomes will focus on discontinuation due to hypotension, bradycardia, renal impairment and HF-exacerbation.Due to the complexity of the statistical analyses, the following section represents the planned principal analyses; some modifications and secondary analyses are likely to emerge during the project. However, a detailed statistical analysis plan will be produced prior to the analysis.Careful initial evaluation will be performed to ensure completeness of data, and to check consistency of the results of the primary analyses for each trial with published reports. Baseline characteristics of patients will be presented separately for each trial and overall. Continuous variables will be presented as mean and standard deviation . Binary and categorical variables will be presented as frequencies and percentages. All analyses will follow the principle of intention to treat as closely as possible. Specifically, we will include all randomized patients with outcome data.P-value.The primary and major secondary outcome will be analyzed using a stratified Cox regression model , with stThe secondary outcomes will be analyzed in the same manner as the primary outcome.Subgroup analyses will be used to assess the effect of beta-blockers in the following pre-specified subgroups: age, diabetes, gender, ejection fraction, atrial fibrillation and etiology of HF. The influence of age and ejection fraction on the effects of beta-blockers will be explored as continuous variables and as clinically relevant categories for other variables. A meta-analysis of interaction estimates will be used to assess whether any improvement depends on baseline LV ejection fraction, blood pressure, and the presence or absence of other concomitant cardiovascular therapies, such as ACE inhibitors, angiotensin receptor blockers, diuretics and digoxin.P-value. To explore the influence of baseline covariates on the primary and major secondary outcomes, we will develop multivariable models using the Cox proportional hazards approach to develop a risk score. In all analyses, continuous variables will be kept continuous, and the nature of their relation to outcome evaluated using fractional polynomials.Multiple imputation for missing data will be used where appropriate as a sensitivity analysis. Dichotomous outcomes will be combined using an inverse variance meta-analysis. Odds ratios and corresponding 95% confidence intervals will be presented, along with the corresponding As noted above, we also plan to perform sensitivity analyses for the exclusion (separately) of the BEST and CAPRICORN trials. If data from the eligible trials are unobtainable, then analysis using a combination of IPD and aggregate data will be considered .For the economic analysis, we propose to perform a cost analysis using standard published information to provide a representative spread of health economic scenarios. Costs of care will be derived from simple drivers like hospital length of stay, medications and other treatments and a cost effectiveness analysis will be carried out based on cost per event avoided . Modeling of cost effectiveness will be carried out based on specific subgroups, such as age, gender and diabetes, in a number of different healthcare models , taking account of the different costs of beta-blockers. An overall population-based cost impact analysis will be derived assessing different levels of uptake of beta-blockers in heart failure with a view to estimating the cost savings of improving beta-blocker utilization.Despite a wealth of information identifying the overall benefits of beta-blockers in HF for morbidity and mortality, prescription rates remain sub-optimal with consequence for both patients and healthcare providers. Current data are limited to those enrolled in the RCTs and lack sufficient statistical power to examine the harm and benefits of treatment in important patient sub-groups. For example, diabetes is present in about 25% of patients enrolled in the larger trials of beta-blockers in HF. The risk of mortality and other complications is higher in diabetics but meta-analysis of published tabular data has suggested that the absolute mortality reduction using beta-blockers may be less in patients with diabetes [Perhaps the most important patient factor that affects prescription of evidence-based therapy is age. In most population-based studies the incidence and prevalence of HF increases with age and the average age of prevalent HF is about 75 years. Most of the trials enrolled patients with a mean age of 60 to 65 years; only SENIORS recruited a population of 70 years or older. The proposed meta-analysis will allow a reliable exploration of any interaction on the effect of beta-blockers with age, albeit limited to the populations recruited in the individual studies. Similarly, most of the trials have enrolled patients with systolic dysfunction data suggest that beta-blockers are effective regardless of baseline renal function ,33. We aFinally, although IPD meta-analyses are considerably more resource-intensive and time-consuming than standard tabular approaches, we believe that only IPD can address the unanswered questions about the use of beta-blockers in HF. Extraction of data from published reports has significant limitations; typically outcomes are described as the number of participants that have died on each intervention over a fixed period, yielding an odds ratio. In comparison, IPD allows a full time-to-event analysis addressing each event from the time of randomization. The hazard ratio obtained provides a more appropriate view of survival and accounts for censored patients. As previously described, IPD also permits robust analyses of subgroups and the ability to estimate the interaction between covariates and treatment effect . The comIn summary, this individual patient-data systematic review and meta-analysis will provide a definitive assessment of the role of beta-blockers in heart failure. The aims of the Collaborative Group are to reduce the burden of morbidity and mortality in heart failure patients and provide clinicians and healthcare agencies with clear guidance on which patients will benefit from beta-blocker therapy.Marcus Flather: Norwich Medical School, University of East Anglia, Norwich, UK.Dipak Kotecha: Clinical Trials and Evaluation Unit, Royal Brompton Hospital/Imperial College London, UK and Monash Centre of Cardiovascular Research and Education in Therapeutics, Monash University, Melbourne, VIC, Australia.Henry Krum: Monash Centre of Cardiovascular Research and Education in Therapeutics, Monash University, Melbourne, VIC, Australia.Luis Manzano: Department of Medicine, Universidad de Alcala, Hospital Universitario Ramon y Cajal, Madrid, Spain.Nicola Williams: Centre for Statistics in Medicine, University of Oxford, Oxford, UK.Douglas Altman: Centre for Statistics in Medicine, University of Oxford, Oxford, UK.Bert Andersson: Department of Cardiology, Sahlgrenska University Hospital, G\u00f6teborg, Sweden.John Cleland: University of Hull, Kingston upon Hull, UK.Andrew Coats: University of East Anglia, Norwich, UK.Mike Domanski: National Heart, Lung and Blood Institute, Bethesda, MD, USA.Guliz Erdem: Clinical Trials & Evaluation Unit, Royal Brompton Hospital/Imperial College London, UK.Marilena Grana: A. Menarini Farmaceutica Internazionale, Firenze, Italy.Per Haglund: AstraZeneca Clinical Information Science, M\u00f6lndal, Sweden.\u00c5ke Hjalmarson: Sahlgrenska University Hospital; G\u00f6teborg, Sweden.Philippe Lechat: Agence Fran\u00e7aise de S\u00e9curit\u00e9 Sanitaire des Produits de Sante, Saint Denis, France.Alain Leizorovicz: Service de Pharmacologie Clinique, Universit\u00e9 de Lyon, Lyon Cedex, France.Mary Ann Lukas: GlaxoSmithKline Cardiovascular and Metabolism Medicine Development Center, Philadelphia, PA, USA.Wilfried Meyer: Merck Serono Global Medical Affairs, Darmstadt, Germany.Milton Packer: UT Southwestern Medical Center, Dallas, TX, USA.Alan Rigby: Academic Cardiology, Castle Hill Hospital, Kingston upon Hull, UK.Giuseppe Rosano: Department of Medical Sciences, IRCCS San Raffaele Pisana, Roma, Italy.Michael Roughton: Clinical Trials and Evaluation Unit, Royal Brompton Hospital/Imperial College London, London, UK.Rosemary Schroyer: GlaxoSmithKline Cardiovascular and Metabolism Medicine Development Center, Philadelphia, PA, USA.Marcelo Shibata: Division of Cardiology, University of Alberta, Edmonton, AB, Canada.Hans Wedel: Nordic School of Public Health, G\u00f6teborg, Sweden.John Wikstrand: Wallenberg Laboratory, G\u00f6teborg University, G\u00f6teborg, Sweden.Thomas von Lueder: Monash Centre of Cardiovascular Research and Education in Therapeutics, Monash University, Melbourne, VIC, Australia.and in memoriam; Philip Poole-Wilson: National Heart and Lung Institute, Imperial College London, London, UK.ACE: Angiotensin converting enzyme; ANZ: Australia/New Zealand Heart Failure Study; ARB: Angiotensin receptor blockers; BB-HF: Beta-Blockers in Heart Failure Collaborative Group; BEST: Beta-Blocker Evaluation Survival Trial; CAPRICORN: Carvedilol Post-Infarct Survival Control in LV Dysfunction Study; CHRISTMAS: Carvedilol Hibernating Reversible Ischaemia Trial: Marker of Success Study; CIBIS I: Cardiac Insufficiency Bisoprolol Study; CIBIS-II: Cardiac Insufficiency Bisoprolol Study II; COPERNICUS: Carvedilol Prospective Randomized Cumulative Survival Study; HF: Heart failure; IPD: Individual Patient Data; LV: Left Ventricular; MDC: Metoprolol in Idiopathic Dilated Cardiomyopathy Study; MERIT-HF: Metoprolol CR/XL Randomised Intervention Trial in Congestive Heart Failure; MI: Myocardial infarction; NYHA: New York Heart Association; RCTs: randomized controlled trials; SENIORS: Study of the Effects of Nebivolol Intervention on Outcomes and Rehospitalisation in Seniors with Heart Failure Study; US-HF: U.S. Carvedilol Heart Failure Study.The majority of the collaborative group have received speaker fees, honoraria or grant support from pharmaceutical companies involved in beta-blocker therapies.The BEST trial was principally sponsored by the US National Heart, Lung and Blood Institute and the Department of Veterans Affairs Cooperative Studies Program. All other trials were funded by pharmaceutical companies.DK participated in the design of the study, manages the collaborative group and performs data management and statistical analysis. LM, GE, HK and MDF participated in the design and coordination of the study. DGA and NW participated in the design of the study and the statistical analysis. All authors drafted, read and approved the final manuscript.Please see attached file for BB-HF data request.Click here for file"} +{"text": "Medical Physics and Engineering was among the first professions to develop and apply e-Learning (e-L). The profession provides excellent background for application of simulations and other e-L materials. The paper describes several layers for e-L development: Programming specific simulations; Building e-L modules; Development of e-L web-based programmes. The paper shows examples from these layers and outlines their specificities. At the end, the newest e-L development (project EMITEL) is briefly introduced and the necessity of a regularly updated list of e-L activities is emphasised. Medical Physics and Engineering (MEP) was among the first professions to develop and apply e-Learning (e-L). An indicator for this is the first international prize in the field (EU Leonardo da Vinci Award) presented to European Medical Imaging Technology (EMIT) Consortium in 2004.During the last 15 years, a number of activities and publications addressed the questions of MEP Education and Training , 3, 4. Te-L is imperative for MEP because it offers quick and easy update of teaching materials \u2013 a very important function for this dynamic profession. This, combined with the fast delivery of the content through Internet, makes e-L materials the first choice for many lecturers;e-L proposes an elegant way to solve the problem through the understanding of complex physics models. Using interactive simulations, computer diagrams or images leads to increased effectiveness of the learning process.Images are specifically important for MEP and e-L provides a cheap and effective means for publishing large number of images (either on CD or through the Web). Additionally e-L can offer a means of image manipulation, which has no analogue in other means of publications.The SEARCH function offered by various e-L materials is another important advantage. This is also imperative for MEP, where currently the specific terms number more than 3,000. This was specially used in the EMITEL Dictionary and Encyclopedia.Finally, the fact that many students from around the world can use the materials through the guidance of most renowned specialists has no analogue in the other educational methods and media.The definitions of e-Learning vary significantly, but perhaps \u201c\u2026 learning that is aided by information and communication technologies\u2026\u201d is one oThe authors certainly support the first view/definition \u2013 that e-L should develop materials, which will increase the pedagogical effectiveness, and then deliver these to students. Only in this case the full power of e-L can be utilised.The development of e-L materials can be presented as a multi-layered process, including the following stages:programming specific simulations;building of e-L modules;development of e-L programmes.These stages most often exist as separate entities, but the programmes will include modules, and simulations can be applied at various stages.Further below this paper shall present some examples of various types of e-L materials. It is impossible to list (or even mention) all existing e-L activities, as this is a very dynamic area and a number of simulations constantly appear or disappear.The simulations are a very effective teaching tool. However, these are very difficult to produce and require good software skills. Also, development of simulations consumes a lot of time. This often requires knowledge of several software packs and skills to present the simulation in a suitable, pedagogical way. One specific problem with simulations is their relatively short life cycle (often less than 5 years). The main reason for this is the upgrade of software platform, what in most cases hampers the performance of the simulation. There is a need for quick dissemination of simulation tools, which could be made through a regularly updated list of available simulations.Among the earliest developments in this field include the Gamma Camera DOS-simulator learning pack ; the PowerLab Systems of AD Instruments; various LabView simulations; the IPEM X-ray Spectrum Processor software, etc. Other existing simulations are described in the special issue on e-Learning of JMEP [Other examples of simulations can be found at the web sites of some manufacturers and professional societies such as AAPM, IPEM, etc. However, the profession lacks a comprehensive list of such tools, which would be of great value for education and training.Building e-L educational modules is probably the most efficient e-L approach. This approach is used in a number of projects and Universities. In principle, one module could cover either educational or training needs (or both) and could have a length of several tens of hours (or days). This allows flexibility by merging the module (or parts of the module) with other pedagogical activities. In this way, the modules are best suited for hybrid delivery of education . Due to this reason, most e-L modules include an option for printing the materials.Most e-L educational modules include only limited interactivity. Perhaps this is due to two reasons: reduced cost for development and increased life cycle. However, these include many images, hyperlinked with text, which increases significantly their educational effectiveness. Also, such modules are very convenient as e-books. Perhaps this is one of the reasons that almost all of them work without tools for students\u2019 assessment.e-L modules also allow flexible application at various levels . An example for such module is Demystifying Biomedical Signals \u2013 a module developed in the Southampton University .www.sprawls.org) and of the IAEA deserve special mention. The IAEA web site includes materials on Protection in: Radiotherapy, Nuclear Medicine and Diagnostic Radiology [Some modules include mainly PowerPoint materials and HTML web pages, but are of extreme importance for the professions. The sets of presentations of the Sprawls Educational Foundation (diology) . These mTypical examples for e-L modules are also the EMERALD and EMIT materials , 11. Thewww.emerald2.eu. The software of the Image database allows interactive off-line manipulation of images like WebCT, Blackboard, etc. These platforms allow easy online administration of the course, but do not always provide new updated teaching materials. It is believed that this will change in future. Good examples are the courses in the University of Adelaide, Australia and the Vanderbilt University, Nashville, USA ,13.Other web-based courses have orientated themselves to the development of specific web-platforms. This approach is very difficult and expensive, but allows the inclusion of custom-built materials and simulations alongside the administration tools of the programme. Attempts in this line have been made, among others, in the University of Gratz, Austria, and Medical University, Plovdiv, Bulgaria ,15. ThesA typical feature of these activities is that the web-based programme can be applied in one University only . Another feature is that these programmes are difficult to maintain. Normally the web site would require constant support from a dedicated experienced webmaster. Due to the recent initiation of a large number of such programmes, an EU project was recently launched . This project is relatThe usefulness of e-L materials has not been disputed, but these require careful assessment. The first Conference to address e-L in Medical Physics was held at International Centre for Theoretical Physics (ICTP), Trieste, during November 2003. Specialists from 26 countries gathered at this Conference to discuss the EMIT e-L materials and the first feedback from the users.Two independent surveys were made at this stage to assess separate tasks on X-ray Diagnostic Radiology and on Ultrasound Imaging. The supervisors reported that in both surveys the students felt better prepared after the e-L task and required less time to complete the task . The students also reported 25-35% improvement in their knowledge after using the e-L tasks, which indicates that the material had been effectively used .Most of the delegates at this Conference agreed on the following main issues related to e-L:e-L is not only suitable, but essential for a dynamic profession like Medical Physics and Engineering;e-L increases enormously the effectiveness of knowledge transfer, but needs to be combined with classical learning \u2013 HYBRID learning;The software platform is crucial for the life cycle of the e-L products (sometimes only an upgrade of the software version can hamper the e-L product);The e-L development team is multi-professional, requiring good professional knowledge, pedagogical experience and IT skills;Producing e-L content is very difficult and costly;e-L development is not less innovative than any other research;The efforts to produce e-Learning materials are often underestimated by students and Universities;There is a need of sound multi-professional forum on e-L.www.emitdictionary.co.uk). It cross-translates Medical Physics terms to/from any of its 15 languages and is soon to be updated to more than 20 languages. This activity will allow colleagues from all over the world to use effectively the existing teaching materials (most of which are in English).The latest development in e-L is the European Medical Imaging Technology e-Encyclopaedia for Lifelong Learning (EMITEL) project, which developed the first e-L multilingual e-Dictionary of Medical Physics and the first e-Encyclopedia of Medical Physics. This large international project includes more than 90 professionals and the first results (the e-Dictionary) are in very advanced stage . It will explain all terms from the Dictionary with articles . Its volume is expected to grow up to above 5 GB. A special Conference will discuss these issues by the end of 2008.EMITEL is based on the results from the previous e-L projects EMERALD, EMERALD II and EMIT. A timeline of their development shows the main stages of these projects . It is ce-Learning has already found a steady presence in Medical Engineering and Physics. A large number of simulations, e-L modules and web-based programmes have been developed and implemented in practice. Some of those, such as the AAPM Virtual Library, the IAEA Training materials and web sites, the EMERALD and EMIT, etc., have reached wide audiences. Many of those materials are still not known therefore these activities need to be urgently listed in order to allow effective exchange of knowledge. Some attempts have been made in this direction \u2013 for example, the book Internet for Radiology, the Global On-Line Medical Physics (GOMP), EMITEL Encyclopedia, several publications, etc. , 19, 20."} +{"text": "History of the World Allergy Organization: In 1951, the leaders in allergy from all over the world came together to form the International Association of Allergology and Clinical Immunology (IAACI). For the next 60 years, the allergy world converged at the IAACI triennial meetings, which became biennial in 2003. The international meetings, originally named the International Congress of Allergology and Clinical Immunology (ICACI), are now the World Allergy Congress (WAC) hosted by the World Allergy Organization (WAO). Everyone who has aspired to have worldwide recognition has played a part in IAACI-WAO. The History of the World Allergy Organization traces the global arc of the allergy field over the past 60 years.The current officers of WAO elected to focus on this rich history, inviting prominent leaders who are interested in being part of this history project to write about their time with IAACI-WAO. This series will be presented in Canc\u00fan, M\u00e9xico as part of the XXII World Allergy Congress . Leading up to the Congress in Canc\u00fan, the World Allergy Organization Journal is presenting segments of the History as part of the \"Notes of Allergy Watchers Series.\" Please enjoy.--Michael A. Kaliner, MDHistorian, and Past-President (2006-2007)World Allergy Organization In September 2003, the World Allergy Organization (WAO)/International Association of Allergology and Clinical Immunology (IAACI) held a scientifically and financially successful Congress in Vancouver, Canada Figure .At this Congress, the name World Allergy Organization was introduced for the first time and used concurrently with the name International Association of Allergology and Clinical Immunology, which it subsequently replaced.The Congress Organizing Committee was co-chaired by Dr. F. Estelle R. Simons and Dr. Michael A. Kaliner. Committee members were Dr. Allen P. Kaplan, WAO President 2000-2003, and Drs. Carlos Baena-Cagnani, G. Walter Canonica, S.G.O Johansson, and Constance Katelaris.The Congress was the second one to be held in Canada since the IAACI began in 1951. It took place in one of the world's most attractive and accessible cities. The Vancouver Convention Centre featured magnificent harbor and mountain views from most windows, highlighting the beauty of the area, including a large nearby urban parkland with towering evergreen trees and fabulous beaches.Dr. Allen Kaplan provided outstanding leadership to the Scientific Program Committee, comprised of Drs. Carlos Baena-Cagnani, Stephen Durham, Takeru Ishikawa, Michael Kaliner, Cas Motala, Johannes Ring, Lanny Rosenwasser, Robert Schellenberg, F. Estelle R. Simons, Daniel Vervloet, and Pakit Vichyanond. The process of developing the Congress was unique in several ways, for example, a needs assessment was conducted, and most of the 270 speakers and moderators in the major sessions representing 6 continents were nominated by their own WAO member societies. More than 800 abstracts were accepted, reflecting cutting-edge allergy and immunology research from laboratories and clinics in 69 nations. The podium and poster sessions included presentations by 30 Congress travel grant winners: 9 young investigators from Europe, 8 from Asia and Australia, 8 from North and South America, and 4 from Africa and the Middle East. Many of these young physicians and scientists are now emerging as leaders in allergy and immunology in their respective countries.Plenary session topics and speakers included the following:The scientific highlights of the meeting were numerous. The Science of Allergy: Initiating MechanismsDrs. William Paul, Andrew Luster, and Kent HayGlassThe Science of Allergy: Effector MechanismsDrs. Dean Metcalfe, Marc Rothenberg, and Barry KayNew Concepts in AsthmaDrs. Stephen Holgate, Richard Martin, and Paul O'ByrneNew Concepts in Allergic Skin DiseaseDrs. Allen Kaplan, Thomas Bieber, and Johannes RingNew Concepts in Allergen ImmunotherapyDrs. Kurt Blaser, Hans-Jorgen Malling, G. Walter Canonica, and Dale UmetsuMeet the Professor sessions featured 16 internationally renowned scientists as chairs and speakers. The topics included the following:Risks and Benefits of Immunotherapy for Allergic Airway DiseaseChair: Dr. Alain de Weck; Speaker: Dr. Stephen DurhamLeukotrienes in AsthmaChair: Dr. Stephen Holgate; Speaker: Dr. K. Frank AustenNew Therapeutic Strategies for Allergic RhinitisChair: Dr. Terumasa Miyamoto; Speaker: Dr. Eli MeltzerTh1/Th2 Polarized ImmunityChair: Dr. William Busse; Speaker: Dr. Patrick HoltCellular and Molecular Aspects of Airway InflammationChair: Dr. Takeshi Fukuda; Speaker: Dr. Barry KayEczema and DermatitisChair: Dr. Albert Oehling; Speaker: Dr. Johannes RingFood AllergyChair: Dr. Pakit Vichyanond; Speaker: Dr. Hugh SampsonAsthma: Inflammatory Markers in Induced SputumChair: Dr. Rajendra Prasad; Speaker: Dr. Frederick HargreaveDebates of the Day , featuring timely topics and 8 well-known professors, proved to be a novel and popular attraction:The 4 Immunotherapy for Asthma? --Dr. Anthony Frew versus Dr. N. Franklin AdkinsonKill the Cat or Buy One? --Dr. Adnan Custovic versus Dr. Thomas Platts-MillsIs the Eosinophil a Player or an Interested Bystander in Asthma? --Dr. Redwan Moqbel versus Dr. Robert Schleimer Chronic Sinusitis: Infection or Allergy? --Dr. Claus Bachert versus Dr. Michael KalinerMore than 30 symposia highlighted the most important allergy/immunology topics of the early 21st century. These included the impact of upper airway allergic inflammation on asthma (WAO Allergy Forum) and advances in occupational allergy . Novel interventions for asthma were featured in symposia on dual inhaled glucocorticoid/longacting beta-agonist therapy, and on anti-IgE therapy. Other symposia focused on the genetics of allergic disease, new trigger factors for allergic disease, interventions to halt the atopic march, and prevention of anaphylaxis recurrences in the community. Additional symposia highlighted the latest information on DNA vaccines, peptide vaccines, and cytokine/chemokine antagonists.The Local Arrangements Subcommittee was co-chaired by Drs. Robert Schellenberg and Donald Stark. Subcommittee members included Drs. Michael Mandl, Ross Chang, Alexander Ferguson, John Dean, H.C. George Wong, Liliane Gendreau-Reid, and Parminder Singh; Gloria Schellenberg, Patricia Stark, Jo-Anne Gillespie, Leslie Chang, Joyce Ferguson, and Narinder Chauhan.aurora borealis was recreated along with a magical backdrop of ice sculptures. At the informal All-Congress WAO Western barbeque held at B.C. Place Stadium, delegates enjoyed authentic Western cuisine and danced the night away. At this event, some attendees wore blue jeans for the first time ever, others panned for gold, and the Royal Canadian Mounted Police made a surprise visit.The social activities highlighted the geographical, historical, and cultural significance of the area. The Opening Ceremony and Welcome Reception focused on the proud heritage and seafaring traditions of the local First Nations peoples, told through music, story, dance, and feasting. For the Congress Banquet, a Northern Lights gala dinner and dance, the Complimentary sightseeing tours were organized for Congress registrants to orient them to the spectacular city of Vancouver, and optional tours were organized to the world-renowned Museum of Anthropology at the University of British Columbia and other Vancouver landmarks.Some delegates traveled the sea-to-sky highway to Whistler Mountain for hiking, gondola rides, canoe rides, and floatplane tours of the glaciers. Others visited Victoria, the beautiful capital city of the province of British Columbia, and a few braved the open Pacific Ocean to go on whale-watching expeditions. Participants were amazed at the ease of access from the Vancouver and Victoria urban areas to the nearby vast and magnificent Canadian wilderness. A number of fortunate delegates experienced one of the world's great panoramic railway journeys, traveling on the Rocky Mountaineer from Vancouver through scenic British Columbia to Banff National Park.Seven years in international planning... 5 wonderful days in our lives.... a lifetime of memories... This sums up the 2003 World Allergy Congress in beautiful Vancouver, Canada."} +{"text": "Recent studies have identified that a higher resting heart rate (RHR) is associated with elevated blood pressure, independent of body fatness, age and ethnicity. However, it is still unclear whether RHR can also be applied as a screening for other risk factors, such as hyperglycemia and dyslipidemia. Thus, the purpose of the presented study was to analyze the association between RHR, lipid profile and fasting glucose in obese children and adolescents.The sample was composed of 180 obese children and adolescents, aged between 7-16 years. Whole-body and segmental body composition were estimated by Dual-energy X-ray absorptiometry. Resting heart rate (RHR) was measured by heart rate monitors. The fasting blood samples were analyzed for serum triglycerides, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and glucose, using the colorimetric method.Fasting glucose, TC, triglycerides, HDL-C, LDL-C and RHR were similar in both genders. The group of obese subjects with a higher RHR presented, at a lower age, higher triglycerides and TC. There was a significant relationship between RHR, triglycerides and TC. In the multivariate model, triglycerides and TC maintained a significant relationship with RHR independent of age, gender, general and trunk adiposity. The ROC curve indicated that RHR has a high potential for screening elevated total cholesterol and triglycerides as well as dyslipidemia.Elevated RHR has the potential to identify subjects at an increased risk of atherosclerosis development. Over the last few decades obesity has reached epidemic proportions and become one of the major public health targets worldwide. Several researches indicate that obesity tracks from childhood to adulthood and constitutes a risk factor in the development of chronic diseases . A high In adults, the use of RHR as screening index for cardiovascular risk has been postulated ,9 and suRecently, Fernandes et al. identifiThus, the purpose of the present study was to analyze the association between RHR, lipid profile and fasting glucose in obese children and adolescents.One hundred and eighty obese children and adolescents , aged between 7-16 years, from Presidente Prudente, western Sao Paulo State, Brazil, were analyzed. The subjects were invited, through television and newspaper advertising, to participate in an intervention program, with physical activity and nutritional orientation, for obese boys and girls .The participants were contacted initially by phone, after which an appointment was made in order to take measurements at the Campus of the Universidade Estadual Paulista - UNESP. Primary obesity diagnosis was made using body mass index (BMI) according to the cutoffs proposed by Cole et al. . After tWhole-body and segmental body composition were estimated by Dual-energy X-ray absorptiometry software version 4.7. Fat free mass (FFM), trunk fat mass (TFM) and percentage of body fatness (%BF) were measured. The DEXA and RHR measurements were made, on the same day, in a temperature-controlled room, in a laboratory at the University.Portable heart rate monitors were used to measure RHR (expressed as beats per minute [beats/min]), which was monitored during two 30-second periods in the sitting position. All measurements were registered after five minutes at rest in a quiet room with a constantly controlled temperature ) were observed. In fact, scientific literature has linked dyslipidemia and arterial hypertension to increased adiposity in children and adolescents % and ele,21. In pIt is well established that the practice of regular physical activity improves the production of superoxide dismutase and nitric oxide ,25 and, Research has shown an increased sympathetic activity in obese individuals -29. SimiThe actual function of some adipokines that affect the insulin binding by blocking the insulin receptor substrates-1 activation, stimulate the lipolysis and contribute to development of dyslipidemia, was recently described . These aIn our study, fasting glucose was not related to RHR. Oda and Kawai identifiA positive aspect of the present study is the analysis of TFM by DEXA. The inclusion of TFM in the multivariate model was important because this adipose tissue is related to the increased release of adipokines related to many pro-inflammatory mechanisms ,3. MoreoOn the other hand, some limitations must be pointed out. The cross-sectional design does not offer support to causality statements and, therefore, prospective studies from childhood to adolescence are necessary to describe more accurately the longitudinal relationship between RHR and dyslipidemia. The absence of inflammatory markers related to oxidative stress and, the absence of insulin measures to screen more clearly the relationship between RHR and glucose metabolism should be considered in future research.In summary, we conclude that increased RHR was significantly associated with dyslipidemia in obese children and adolescents and that elevated RHR offers potential to screen subjects at an increased risk of atherosclerosis development. However, longitudinal and epidemiological surveys should be carried out to develop optimal cutoff values for RHR in pediatric populations.RHR: resting heart rate; BMI: body mass index; DEXA: dual-energy X-ray absortometry; FFM: fat free mass; TFM: trunk fat mass; %BF: body fat percentage; Beats/min: beats per minute; TC: total cholesterol; HDL-C: high-density lipoprotein cholesterol; LDL-C: low-density lipoprotein cholesterol; TC: total cholesterol; ROC: receiver operation characteristic; SD: standard-deviation.The authors declare that they have no competing interests.RAF: (1) conception and design of the study, (2) acquisition, analysis and interpretation of data, (3) draft of the article and selection of manuscripts to discuss the results, PAM, LSS, SAU, BMA, KNB and JSC: (1) Acquisition, analysis and interpretation of data, (2) draft of the article and selection of manuscripts to discuss the results, IFFJ and JPJS: (1) conception and design of the study (2) review and approval of the final version to be submitted. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2431/12/5/prepub"} +{"text": "At the same time, nutrition-related health conditions, such as obesity and type 2 diabetes, are increasing in Mexican-origin youth. Risk factors for obesity and type 2 diabetes are more common in Mexican-origin children and include increased intakes of energy-dense and nutrient-poor foods. This study assessed the relationship between children's experience of food insecurity and nutrient intake from food and beverages among Mexican-origin children (age 6-11 y) who resided in Texas border promotora-researchers. All survey (demographics and nine-item child food security measure) and 24-hour dietary recall data were collected in Spanish. Dietary data were collected in person on three occasions using a multiple-pass approach; nutrient intakes were calculated with NDS-R software. Separate multiple regression models were individually fitted for total energy, protein, dietary fiber, calcium, vitamin D, potassium, sodium, Vitamin C, and percentage of calories from fat and added sugars.Baseline data from 50 Mexican-origin children were collected in the home by trained Thirty-two children (64%) reported low or very low food security. Few children met the recommendations for calcium, dietary fiber, and sodium; and none for potassium or vitamin D. Weekend intake was lower than weekday for calcium, vitamin D, potassium, and vitamin C; and higher for percent of calories from fat. Three-day average dietary intakes of total calories, protein, and percent of calories from added sugars increased with declining food security status. Very low food security was associated with greater intakes of total energy, calcium, and percentage of calories from fat and added sugar.This paper not only emphasizes the alarming rates of food insecurity for this Hispanic subgroup, but describes the associations for food insecurity and diet among this sample of Mexican-origin children. Child-reported food insecurity situations could serve as a screen for nutrition problems in children. Further, the National School Lunch and School Breakfast Programs, which play a major beneficial role in children's weekday intakes, may not be enough to keep pace with the nutritional needs of low and very low food secure Mexican-origin children. These settlements are occupied by a growing population of people who share a similar Mexican heritage, language, and socioeconomic standing and who have unacceptably high rates of poverty, adult and childhood obesity, and food insecurity 2. Appropriate Centers for Disease Control and Prevention (CDC) BMI-for-age-and-sex growth charts were used to classify each child's BMI status as underweight (< 5th percentile), healthy weight (5th percentile to < 85th percentile), overweight (85th percentile to < 95th percentile), or obese (\u226595th percentile) [centile) ,40.Children's food security was assessed using the nine-item child food security measure developed by Connell and colleagues [promotora-researcher whether they experienced each of the nine items during the last three months (see Table lleagues . Pilot tpromotora-researcher). In most cases, the mother observed and assisted the children if necessary. Dietary intake training for the interviewers included review of all protocols and scripts, modeling of interviewing, practice interviews with children similar in age to the study participants, use of tools for portion-size estimation, quality control, and focus on children's reporting of food items. The first recall occurred during the first in-home visit, and the second and third recalls were collected in the home during the second and third visits (within two weeks of the first visit). Detailed information on food and beverage consumption, including description, brand name, location of preparation and consumption, and preparation method during the previous day was collected using standardized protocols following a modification of the multiple-pass interview technique of the Nutrition Data System for Research (NDS-R) [Three 24-hour (previous day) dietary recalls occurring on randomly selected, nonconsecutive days were collected in the home from each participant child by the same interviewer ( (NDS-R) . Data we (NDS-R) , and the (NDS-R) . ChildreAll analyses were performed using Stata Statistical Software: Release 11 . Descriptive statistics were calculated for each child's baseline characteristics, BMI status, food security status, and nutrient intake. Wilcoxon Signed-Rank Test was used to compare weekend and weekday nutrient intake by level of food security. Non-parametric test for trend across ordered groups of food security was used to examine each nutrient. Separate multiple regression models with robust (White-corrected) Standard errors (SEs), were individually fitted for total energy , protein (g), dietary fiber (g), calcium (mg), vitamin D (mcg), potassium (mg), sodium (mg), Vitamin C (mg), percentage of calories from fat, percentage of calories from added sugars, and percentage of calories from saturated fat. All models included sex, age, country of birth, BMI status, and food security status as independent variables. These variables were selected based on their documented association with dietary intake.n = 18) or very low food security (n = 14). Although BMI status was not significantly associated with food-security status, 48 percent of the sample was measured as being of healthy weight and reporting low or very low food security. Nutrient intake and dietary recommendations for the entire sample and nutrient intake by food-security status are shown in Table n = 14) met the recommendations for calcium, none for potassium or vitamin D, 10% (n = 5) for dietary fiber, and 6% (n = 3) for sodium (data not shown). Although all children exceeded the recommendation for protein, as a percent of total calories, protein intake ranged from 11.8% to 22% (data not shown). Weekend intakes for calcium, vitamin D, potassium, and vitamin C were significantly lower than weekday consumption, and percentage of calories from fat, and combined percentage from fat and added sugar (data not shown) were significantly higher on weekends than weekdays. Children who were identified with low food security consumed significantly less calcium and vitamin D on weekends, compared with weekdays, and very low food-security children consumed a greater percentage of calories from fat on weekends than weekdays. Three-day average intake for total energy, protein, percentage of calories from added sugar, and percentage of calories from saturated fat demonstrated a significant and positive trend (indicating greater intake) with reduced food-security status. The same positive trend was observed for weekend intake of total energy, and for weekday intake of percentage of calories from added sugars and saturated fat.Table p = 0.032), low , and very low food security were associated with increased intake as a percentage of calories from combined fat and added sugar. In addition, increased age was associated with lower intakes of calcium and vitamin D; being born in Mexico with greater intake of sodium; and being female with lower calcium intake. BMI status was not associated with any of the nutrients.The multiple regression results, presented in Table colonias [Previous work has recognized the relationship between food security and children's diets ,35,37, bcolonias than previous estimates suggested. For instance, national data from 2009 indicated that 18.7% of Hispanic households, regardless of race or country of origin, had food-insecure children [colonias reported that 49% of all households and 61.8% of households with children were classified as child food insecure [Results present additional evidence that food insecurity is more prevalent among Mexican-origin children in Texas border insecure . In the insecure . In addiinsecure , and is insecure .This paper not only emphasizes the alarming rates of food insecurity for this Hispanic subgroup, but describes the associations for food insecurity and diet among this sample of Mexican-origin children. Such findings have implications at a regional and national level, as the Mexican-origin population continues to grow along the border and in new destination communities ,45. Immicolonias. This population is of increasing national importance because such colonias can be considered an archetype for the new-destination Mexican immigrant communities that are now found in great numbers throughout the U.S. Second, to our knowledge, this is the first study that uses children's report of their food-insecurity experiences in the past three months to describe food-security status, which is preferable and reduces the cognitive burden placed on respondents by the conventional twelve-month time period [The present study has several particular strengths. First, this is a study of hard-to-reach Mexican-origin children in border e period . As suche period ,35,37, te period -51.colonias in the Texas border region, which limits our ability to generalize these results. Future work should focus on expanding our understanding of seasonal variation in the frequency and duration of children's experiences of food insecurity.There are several limitations to this study that warrant mention. Data were collected during one season of the year, which limits our ability to describe seasonal variation in dietary intake or food-security status or to make causal inferences. This could have important implications for times of year when children are unable to participate in school nutrition programs, such as during the summer or holidays. Although the three-month time frame was much better than asking about food security experiences in the last 12 months, this study did not collect data on frequency or duration of food insecurity situations. This limits our ability to differentiate between acute and chronic food insecurity. An additional limitation is an absence of data on food coping strategies employed by children to help manage food resources . Finallycolonias, food insecurity may be an ongoing reality. The prevalence of low and very low food security in this border area is alarming, despite the participation of all study participants in the School Breakfast and National School Lunch Programs. The high prevalence of low and very low food security among these children is especially troubling given the importance of good nutrition on optimal growth, function, and health [Despite these limitations, the results of this study further the knowledge of children's experiences of low and very low food security and the association of food security status with children's dietary intake. The Mexican-origin population is rapidly expanding throughout the United States; record numbers of individuals and families are experiencing food insecurity, and for children living in rural or underserved areas such as the d health ,20. Yound health , and ared health . In thisThe authors declare that they have no competing interests.JRS designed the study, and worked on the development of the instrument and the protocol for collection of data. JRS, CN, CMJ, and WRD wrote the first draft of the paper. JRS, CN, CMJ, and WRD read and approved the final manuscript.JRS is Professor of Social and Behavioral Health and Director of the Program for Research in Nutrition and Health Disparities; CN is a Graduate Research Assistant; CMJ was a Research Associate; and WRD is Assistant Professor of Social and Behavioral Health.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2431/12/16/prepub"} +{"text": "The challenge features a set of ten unlicensed breast cancer inventions that have commercial viability and the potential to improve public health. The primary goal is to stimulate the creation of start-up businesses based on these inventions. Teams \u2013 made up of students led by established entrepreneurs \u2013 are competing to devise business plans; their efforts will be judged by an expert panel, including, among many other leaders in the pharmaceutical industry, George F. Tidmarsh, MD, PhD, CEO of La Jolla Pharmaceutical Co. and Consulting Editor of Disease Models & Mechanisms (DMM), Michael King Holly, Pharm. D, Senior Vice President of Quintiles Innovation, and Katherine Bowdish, PhD, Vice President R&D and Head of Sunrise, Sanofi.In September 2013, the Avon Foundation, the NIH-funded National Cancer Institute (NCI) and The Center for Advancing Innovation announced their partnership to create the first Breast Cancer Start-up Challenge emphasized that the challenge is designed to accelerate and increase the volume of breast cancer inventions in development. Moreover, they hope to \u201cspur economic growth and provide universities with a platform to develop their entrepreneurship-learning portfolios\u201d.Marc Hurlbert, Executive Director of the Avon Foundation Breast Cancer Crusade (http://www.breastcancerstartupchallenge.com/teams-participating.html. Challenge teams include but are not limited to students from the medical, business and/or law schools of Wake Forest University, University of North Carolina, Harvard, NYU, Rutgers, Ohio State, and the Icahn School of Medicine at Mount Sinai. The challenge has also attracted team members from outside the USA, including India, UK, Canada, New Zealand and The Netherlands. Each team must incorporate at least four disciplines: legal, medical/scientific, business and a seasoned entrepreneur. Illustrating the breadth of experience and expertise, the cumulative number of years of experience on all teams participating includes:A total of 200 teams received invitations to enter the contest, and 46 teams have been accepted with 431 people cumulatively participating within teams: 791 in start-ups1523 in science/medicine839 in consulting in a relevant field1203 in life sciences business operations.Tom Stackhouse, Associate Director of the Technology Transfer Center at the National Cancer Institute, said: \u201cWe are excited that so many high-quality teams have come together for this challenge. We are looking forward to seeing the outcome\u201d.DMM will provide readers with updates during each phase of the challenge, and will profile the winning team at the end of the contest, scheduled for June 27, 2014."} +{"text": "Surgical treatment of Jehovah\u2019s witnesses, even if elective, is a difficult situation for the surgeon from a clinical, deontological and ethical point of view. Any lesion or the death of the patient can result in civil and penal consequences for the surgeon and the anesthetist.The aim of the present study was to present our experience regarding operative mortality and early clinical outcome in the surgical treatment of Jehovah\u2019s witnesses compared to non-Jehovah's witness patients.The research was structured as a retrospective cohort study. The total cohort analyzed was 80 subjects recruited from the Veneto Region, North-east Italy. Patients were divided into two groups:Group 1, n=40, Jehovah's witness patients undergoing colorectal elective surgery.Group 2, n=40, non-Jehovah's witness patients matched according to sex, age, type of surgery.Informed consent was collected for each patient. Preoperative, intraoperative and postoperative conditions were analyzed. In particular, in the preoperative phase cardiovascular and hematologic conditions (hematocrit - Hk - and hemoglobin - Hb) were assessed; in the intraoperative and postoperative phases bleeding with severe anemization, length of stay in the intensive care unit and in our ward, re-admission rate, and mortality were considered.All patients survived. No blood transfusions were administered to Jehovah\u2019s witness patients. Severe complications and mortality rate were similar in the two groups. In particular, intra and postoperative bleeding, the need for intubation/resuscitation, and days of hospitalization were not significantly different (p> 0.05) in the two groups of patients. No malpractice claim was ruled.An accurate selection of patients, the minimization of perioperative blood loss, perioperative collection of autologous blood and meticulous surgical techniques could be responsible for the low rate of complications observed in both groups and the absence of claims of medical liability."} +{"text": "Integrated care has moved from the small niche it traditionally occupied in academia, accessible only to experts in the field and applied merely on a project specific or pilot effort basis, now to the radar of politicians and health system planners the world over.More than a buzzword for the 21st century, coordinated/integrated health services delivery is a necessity. From changing demographics and increasing chronicity to the persisting threat of communicable diseases, coupled with modern technologies, rising patient expectations and a perpetual context of fiscal constraints, new and innovative approaches to the delivery of health care, that ensure high-quality services which are efficient in their provision and delivered according to an individual's needs, must be given the utmost priority.Strengthening the coordination/integration of care is ultimately best viewed as a means, rather than an end in itself, for improved health outcomes. The activity in this area can be credited to this presumed causal relation. The vast number of initiatives to design and implement integrated care programmes across countries in the WHO European Region is astounding. These examples extend from local initiatives like integrated care pilots in North West London in the United Kingdom to regioStill, there remains a long way ahead to transform services to people-centred care: lack of clearly defined and measurable aims, consistent communication strategies or participatory approaches in developing and implementing integrated care are only a few of the shortcomings that may lead to sub-optimal outcomes or present challenges in order to implement efforts sustainably and at scale. Ensuring system-wide implementation is often further hampered by ambiguous incentive structures, a lack of adequately trained professionals and/or outdated legal frameworks.from practice and for practice. Reforming a health system to meet the challenges of the 21st century is no small feat and it is only with the many different attempts to integrating care that we have come to understand the complexities of health system. As Goodwin recently acknowledged [The variability in approaches is in itself a testament to the diversity and flexibility of integrated care \u2013 a characteristic that is also its strong point, as integrated care has never been intended as a mere theoretical concept, but was rather developed owledged , there iowledged .As the interest for integrated care has grown, so has the attention of governments, the World Health Organization, the European Union and other international institutions in this topic. At the WHO Regional Office for Europe, attention is drawn to the vision of Health 2020 for imprcoordinated/integrated health services delivery and how can it be operationalised to lead to health system strengthening?What is This will be responded to within a concept note, synthesising the existing literature and evidence, and outlining WHO Europe's definition of coordinated/integrated health services delivery as the design, management and delivery of health services such that people receive and perceive a continuum of health promotion, health protection and disease prevention services, as well as diagnosis, treatment, long-term care, rehabilitation and palliative care services through the different levels and sites of care within the health system and according to their needs. The uniqueness of WHO's mandate permeates this definition, which includes public health as a vital part of the health system, and puts people, not necessarily patients, into the centre of attention. In operationalising this definition, intersectoral actions towards including social services, educational sector and legal frameworks are needed throughout the integration process.What are the experiences of Member States with implementing more integrated care and how do the lessons from these implementation efforts apply to other contexts?A second pillar collects experiences from across the WHO European Region, with its 53 member states, by means of an open call for initiatives and in-depth country case studies. While the former will draw a picture of what is already happening and where, the latter will identify the arenas for action necessary to create sustainable and equitable systems change towards coordinated/integrated health services delivery. As outlined above, successful examples like the Eastern Lithuanian Cardiology Programme , the \u2018HeHow can policy-makers, providers, patients and communities lead and manage transformations towards more coordinated/integrated health services delivery?Finally, the case studies and country experiences, as well as the existing knowledge captured in the concept note, will inform recommendations and policy options to manage and implement sustainable change. By creating an environment conducive to innovative approaches, it is anticipated that a change in values and cultures will also take place, from the primary care setting across the boundaries to hospitals and long-term care facilities, in order to overcome barriers and enable a truly holistic approach towards people-centred services.Challenging the status quo is never easy. But as our Member States have shown, changing the delivery of care to provide more coordinated/integrated health services is a needed effort towards high-quality, sustainable, people-centred health systems. And it can be done. The momentum this agenda has garnered is merited to its applicability given the current context of health and society. Nevertheless, much work remains. We must continue to expand our thinking and experiences in this field, overcoming limitations of small-scale efforts, responding to concerns of sustainability, and ultimately, reaching a level of sophistication that enables the coordination/integration of services to be truly tailored to the individual and not restricted, for example, by geography, disease types or privileged groups of society. That is, in continuing to transform our health services towards more coordinated/integrated care, ultimately, each nation, region, community, family and individual may realise their full health potential.Just this past October, the WHO Regional Office for Europe officially launched the roadmap to develop a Framework for Action towards coordinated/integrated health services delivery at the 5th Tallinn Charter Anniversary WHO High-Level Meeting in Tallinn, Estonia. This framework follows the call from Member States for contextualised, evidence-based policy options to enable system-wide changes and the need for tools to implement these changes. The framework is aligned with other WHO global initiatives in this area and involves a wide range of stakeholders including national representatives from countries across Member States and international experts, among others, to solicit their input through various consultations and feedback loops. The way forward was defined in the aforementioned roadmap document, which outlines three pillars for action. These pillars correspond to three topical questions:The WHO Regional Office for Europe new health policy, Health 2020 , calls fNote: As a first effort to consolidate experiences, we invite the exchange of practices to create more coordinated/integrated services from across the WHO European Region through an open call for initiatives. Whether a local effort to minimise fragmentation of care in a given centre, or a regionally or nationally planned initiative linking providers across sites and levels of care \u2013 whichever the scale, shape and form \u2013 your support in capturing these efforts is needed. For more information on this open call for practices and to fill in the simple electronic questionnaire to do so \u2013 as a manager, policy-maker, provider or participant in an initiative \u2013 we invite you to visit our website at: www.euro.who.int/en/what-we-do/health-topics/Health-systems/health-service-deliveryEnglish:http://www.euro.who.int/en/health-topics/Health-systems/health-service-delivery/news/news/2013/11/call-for-submission-of-initiatives-towards-the-coordinationintegration-of-health-services-delivery-cihsd-in-the-who-european-region/_recacheRussian:http://www.euro.who.int/ru/health-topics/Health-systems/health-service-delivery/news/news/2013/11/call-for-submission-of-initiatives-towards-the-coordinationintegration-of-health-services-delivery-cihsd-in-the-who-european-region/_recachecihsd@euro.who.intTo send questions, feedback or input, please contact"} +{"text": "AbstractScutellistacaerulea was imported and released in Nelson, New Zealand, for the biological control of pest scale insects. It was thought to have failed to establish, and is therefore currently considered to be absent from the New Zealand fauna. On 17 April 2013, a live specimen was captured in the wild in Auckland.In 1921, Scutellistacaerulea was released in Nelson, New Zealand, for biological control of pest scale insects, but apparently failed to establish (Scutellistacyanea). It is not considered to be present in New Zealand, and is not listed on the New Zealand Organisms Register (NZOR). However, on 17 April 2013, I captured a single live female in the wild, in the grounds of the Tamaki Campus of the University of Auckland Scutellistacyanea Motschulsky, 1859Type status:Other material. Occurrence: recordedBy: Stephen Thorpe; individualCount: 1; sex: female; Location: country: New Zealand; verbatimLocality: Tamaki Campus of University of Auckland; verbatimLatitude: 36.8816119078S; verbatimLongitude: 174.8531936109E; Event: eventDate: 17 April 2013; Record Level: institutionCode: Auckland MuseumScutellistacaerulea be added to the New Zealand Organisms Register (NZOR) as exotic, present in the wild. It is unclear if the new specimen represents a descendant of the original stock released in Nelson in 1921, or a new incursion from overseas. The balance of evidence favours the latter hypothesis, since Auckland is far from Nelson, and the species has not been seen in N.Z. for nearly a century.The specimen was identified using"} +{"text": "Low socioeconomic status (SES) is associated with mortality in several populations. SES measures, such as education and income, may operate through different pathways. However, the independent effect of each measure mutually adjusting for the effect of other SES measures is not clear. The association between poverty-income ratio (PIR) and education and all-cause mortality among 15,646 adults, aged >20 years, who participated in the Third National Health and Nutrition Examination Survey in the USA, was examined. The lower PIR quartiles and less than high school education were positively associated with all-cause mortality in initial models adjusting for the demographic, lifestyle and clinical risk factors. After additional adjustment for education, the lower PIR quartiles were still significantly associated with all-cause mortality. The multivariable odds ratio (OR) [95% confidence interval (CI)] of all-cause mortality comparing the lowest to the highest quartile of PIR was 2.11 . In contrast, after additional adjustment for income, education was no longer associated with all-cause mortality [multivariable OR (95% CI) of all-cause mortality comparing less than high school to more than high school education was 1.05 ]. The results suggest that income may be a stronger predictor of mortality than education, and narrowing the income differentials may reduce the health disparities. Socioeconomic inequalities in health are a major public-health concern. Epidemiological studies have shown that socioeconomic status (SES) is associated with mortality in several populations -10. AlthThe Third National Health and Nutrition Examination Survey (NHANES III) collected data on a nationwide probability sample of the civilian non-institutionalized US population. Detailed descriptions of the complex survey design, interviewing procedures, and physical examinations conducted have been published before and are available online .Information on mortality status was available for 18,800 participants aged \u226520 years. We further excluded those with missing data on poverty-income ratio , education (n=96), and other variables included in the multivariable model, leaving 15,646 available for current analysis. Compared to those who were included in the final analysis, those who were excluded were: older, more likely to be female, primary or below educated, to have lower PIR, less likely to be non-Hispanic whites, consume alcohol, physically active, and had higher levels of mean arterial blood pressure (MABP) .The main outcome of interest\u2014all-cause mortality\u2014was recorded from the NHANES III-linked mortality file provided by the National Center for Health Statistics (NCHS). The participants were followed up for mortality for up to 12 years from 1988-1994 through 31 December 2006. Ascertainment of mortality was based upon a probabilistic match between the NHANES III and the death certificate records of the National Death Index. Education and PIR were chosen as indicators of SES. PIR was computed as a ratio of the mid-point of the observed family-income category to the family's appropriate poverty threshold set by the US Census Bureau in a given calendar year. The educational status based on completed years of education was categorized into high school graduate .We first examined the association between PIR and all-cause mortality and subsequently examined the association between education and all-cause mortality. To examine the independent effect of education and income on all-cause mortality, we used three logistic regression models: (a) the age, sex-adjusted model; (b) the multivariable-adjusted model 1, additionally adjusting for race-ethnicity, marital status, smoking, alcohol intake, physical activity, MABP, body mass index, high-density lipoprotein cholesterol, and (c) the multivariable-adjusted model 2, adjusting for all variables in the multivariable model 1 plus mutual adjustment of other SES measures (education in models of PIR and PIR in models of education). The trends in the odds ratio (OR) of all-cause mortality across the categories of PIR and education were tested by modelling each SES indicator category as an ordinal variable in the corresponding multivariable model. In subgroup analyses, we examined the association among education, income, and all-cause mortality stratified by race-ethnicity and gender using the multivariable model 2. All analyses were weighted to account for unequal probabilities of selection, oversampling, and non-response using the SUDAAN software (version 8.0) and the SAS software (version 9.2) .Participants signed an informed consent form before interview in the home. The ethical approval was obtained from the Institutional Review Board of the National Center for Health Statistics of the Centers for Disease Control and Prevention.In a contemporary, multi-ethnic sample of US adults, we found that the lower PIR quartiles were positively associated with all-cause mortality, independent of the demographic, lifestyle and clinical risk factors. This association was persistent after additional adjustment for education and was consistently present in subgroups of gender and race-ethnicity. In contrast, education was not associated with all-cause mortality after additional adjustment for income.The majority of the US studies that examined the association between the SES and mortality have used education ,6 or incAlthough the strengths of the study include its large sample-size, rigorous methodology, and rich information on covariates, our study has some limitations. First, since the SES variables were assessed only once as part of the NHANES III, the effect of changes in income over time on mortality could not be studied. Second, the possibility of residual confounding due to measurement error resulting from broad categorization of covariates, for example smoking and drinking, cannot be excluded.In a nationally-representative sample of US adults, we found that lower income was positively associated with mortality, independent of the demographic, lifestyle and clinical risk factors, and this association persisted after additional adjustment for education. In contrast, after additional adjustment for income, lower education was not associated with mortality. Our results suggest that income may be a stronger predictor of mortality than education, and narrowing the income differentials may reduce health disparities.The study was funded by an American Heart Association National Clinical Research program grant (AS)."} +{"text": "There was an error in the third from last statement in the Funding section.The correct sentence is \"This research was partially supported by grants from Academia Sinica, Taiwan, and from the National Science Council, Taiwan (NSC101-2311-B-001-002) to MHC, the NIH (U54GM094662 to SCA and AI095382 to RHC) and the Albert Einstein Cancer Center (CA013330).\""} +{"text": "Multidrug resistant microorganisms (MDROs) constitute a serious threat. Strategies to reduce incidence are based on reducing both emergence and transmission. e-health, defined as intensive use of information and communication technologies, can be a major part of strategies to reduce the rate of MDROs.Aiming improvement of the quality of antibiotic use based on the e-health concept in a 280-bed, paper-free, general hospital, Infection Control Committee (ICC), along with Quality and Antibiotics Committees (Q&A) and IT team, has been working on implementation of new tools at several levels. On the electronic medical record, a new template for antimicrobial prescription has been created and is in use since January, conditioning automatically the prescription in terms of context and duration . If there is a disagreement between prescriptions and protocols, physicians may proceed, but an electronic justification must be fulfilled and sent to the ICC and Q&A Committees, which, in turn interact with prescribers both by e-mail and phone call, thus building up an antibiotic stewardship. This is complemented by e-pub of antimicrobial protocols on the hospital intranet and by release of data on antibiotic use.Data analysis is still on process.The authors hope that the use of e-health on antibiotic management will improve the quality of care and thus reducing MDROs emergence.None declared."} +{"text": "We aimed to analyze outcomes of early and delayed laparoscopic cholecystectomy in the elderly in our General Surgery Division.We analyzed 114 LC performed from the 1st of January 2008 to the 31st of December 2012 in our General Surgery division: 67 LC were performed for gallbladder stones and 47 for acute cholecystitis.Comparison between Ordinary and Emergency groups showed that drain placement and post-operative hospital stay were significatively different. There were no significative differences between Early Laparoscopic Emergency Cholecystectomy (E-ELC) and Delayed Laparoscopic Emergency Cholecystectomy (D-ELC). There weren't any differences about Team's evaluation.We consider LC a safe and effective treatment for cholelitiasis and acute cholecystitis in Ordinary and Emergency setting, also in the elderly. We also demonstrate that, in our experience, LC for AC is feasible as well. Laparoscopic cholecystectomy (LC) represents the gold standard treatment for cholelithiasis.Its application gradually extended to acute cholecystitis (AC) also in the elderly. We aimed to compare outcomes of the University Section of General Surgery in \"San Luigi Gonzaga\" Hospital of Orbassano (Turin) with literature, evaluating timing and technique of early or delayed laparoscopic cholecystectomy in the management of acute cholecystitis in elderly patients.From the 1st of January 2008 to the 31st of December 2012, 114 LC were performed at the University Section of General Surgery in elderly patients Age > 65 yrs): 67 for gallbladder stones and 47 for acute cholecystitis. The diagnosis of cholecystitis and gallbladder stones was performed basing on general conditions, physical examination, laboratory exams, radiologic findings and sepsis score. For the study we also considered: total hospital stay, time before and after surgery, duration and kind of operation, conversion to open procedure, drain and final pathological results. We excluded 29 patients from the study . We didn't exclude ASA III and ASA IV patients: in these patients we used abdominal pressure not superior of 10 mmHg [ yrs: 67 version 2.6.2), and a p value of less than 0.01 was considered indicative of statistical significance.Statistical proportions related to the analyzed dichotomic variables, for both E-ELC and D-ELC were compared using Chi-square test and Fisher's exact test. Continuous variables like age distribution, post-operative hospital stay time, surgery duration and several haematochemical characteristics were expressed as average (range) and analyzed using the Mann-Witney U test. Patient distribution according to different surgical teams was confirmed. All statistical analyses were performed using R software (In our experience, the comparison between Ordinary and Emergency Group was no statistically significant about blood test values and ultrasonographic evidence Table .We analyzed E-ELC and D-ELC data without finding any statistically significant difference in the elderly, except for the full hospital stay duration, which was longer for D-ELC patients Table . OperatiIn agreement with literature -10, we cThe authors declare that they have no competing interests.AGF: conception and design, interpretration of data, given final approval of the version to be published.SE: conception and design, interpretration of data, given final approval of the version to be published.MS: acquisition of data, drafting the manuscript, given final approval of the version to be published.AF: acquisition of data, drafting the manuscript, given final approval of the version to be published.SC: acquisition of data, drafting the manuscript, given the final approval of the version to be published.GP: acquisition of data, drafting the manuscript, given the final approval of the version to be published.SM: critical revision, interpretation of data, given final approval of the version to be publishedVM: critical revision, interpretation of data, given final approval of the version to be published"} +{"text": "In Zhejiang Province, there are several highly developed cities near the coast and several relatively under-developed mountain areas. Analysis of the composition of bacteria isolated from patients as well as their antibiotic resistance profile from various areas of this province, and tracing of such data year-by-year, will help to delineate the bacterial resistance profile of these areas and to understand how the stage of socio-economical development impacts on the composition of clinical micro-flora and their resistance profile.Enterobacteriaceae, from 2000 to 2009 in Zhejiang Province, China, Enterobacteriaceae isolated from 15 hospitals located in different regions of the province were surveyed.In order to investigate variation in resistance rates and isolation rates of Enterobacteriaceae isolated increased more than 20-fold from 2000 to 2009. Among the Enterobacteriaceae, Escherichia coli and Klebsiella pneumoniae were the dominant isolates. The percentage of E. coli and K. pneumoniae that produced detectable extended-spectrum \u03b2-lactamases (ESBLs) increased from 2000 to 2007, and then declined slightly in 2008 and 2009. The percentages of K. pneumoniae and E. coli that were resistant to ceftazidime increased sharply from 2000 to 2009. There were remarkable increases in the carbapenem resistant rates during the decade, but they were much higher for the isolates from the developed cities than from the rural areas. In 2002, carbapenem-resistant E. coli was first found in Hangzhou, one of the highly developed cities in Zhejiang Province. By 2009, carbapenem-resistant bacteria were found for all species of Enterobacteriaceae surveyed in almost all areas of the province, although they were more frequently identified in developed areas than in rural areas.The total numbers of the Much restrictive actions have to be taken in terms of rational use of antibiotics and nosocomial control to prevent the further spread of the drug-resistant pathogens. The reformation and opening up of China in the last two decades of the 20th century was accompanied by socio-economic development and a gradual improvement in the standard of living. The life style of the Chinese people, especially people living in the relatively developed areas of China, has undergone significant changes. Such changes have had a profound impact on micro-flora and resistant bacteria both in the environment as well as in clinical settings. One of the most important factors that contributed to these changes in micro-flora and to the resistance profile of bacteria was the abuse of antibiotics. Thus, antibiotics have been widely used and, in addition, over-doses of various antibiotics have been administered for healthcare and in farming where they are used as animal feed additives. The abuse of antibiotics in medical settings has directly impacted the resistance profile of clinically isolated bacteria. Moreover, antibiotics used for crop cultivation and/or in animal feeds not only alter micro-flora in the environment, which in turn impact on clinical micro-flora, but also increase the chance of exposure of humans to these antibiotics.Zhejiang Province, which has a population of over 46 million, is located on the east coast of China and is the most economically developed province in China. While it is one of the largest areas in China for farming and aquaculture, this province also has the most highly developed medical system, with the highest medical consumption per capita among all of the provinces in China. However, the level of development among different areas even within this same province is not equal. There are several highly developed cities near the coast such as Hangzhou , Ningbo, Wenzhou, and Shaoxing, as well as several relatively under-developed mountain areas such as Lishui and Quzhou. Analysis of the composition of bacteria isolated from patients as well as their antibiotic resistance profile from various areas of this province, and tracing of such data year-by-year, will help to delineate the bacterial resistance profile of these areas and to understand how the stage of socio-economical development impacts on the composition of clinical micro-flora and their resistance profile.Enterobacteriaceae, are the most prevalent. The CHINET 2008 surveillance of bacterial resistance showed that Enterobacteriaceae account for 52% of clinical gram-negative bacteria, and that Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Proteus mirabilis, Citrobacter freundii, and Serratia marcescens were the most commonly isolated Enterobacteriaceae (E. coli accounts for 26.4\u201327.6% of gram-negative bacteria followed by Klebsiella spp. (13.8\u201319.6%) and Enterobacter spp. (4.7\u20135.8%) (Enterobacteriaceae infection. The Enterobacteriaceae that predominantly produce extended-spectrum \u03b2-lactamases (ESBLs) are E. coli and K. pneumoniae, and ESBL production was detected in close to 56.2 and 43.6%, respectively of these bacteria, consistent with previous studies , Zhejiang Provincial People's Hospital (Hangzhou), Zhejiang Provincial Hospital of Traditional Chinese Medicine (Hangzhou), Hangzhou First people's Hospital (Hangzhou), Hangzhou Third people's Hospital (Hangzhou), Zhuji People's Hospital of Zhejiang Province (Shaoxing); the First Hospital of Jiaxing (Jiaxing), Huzhou Central Hospital (Huzhou), Ningbo First Hospital (Ningbo), Quzhou Central Hospital (Quzhou), Dongyang People's Hospital (Jinhua), Lishui Central Hospital (Lishui), Taizhou Hospital of Zhejiang Province (Taizhou), Zhoushan Hospital (Zhoushan), the Second Affiliated Hospital of Wenzhou Medical College (Wenzhou). Isolates were collected from aseptically obtained body fluid such as blood, urine, pleural fluid, and ascites from both in-patients and out-patients during January 2000\u2013December 2009.E. coli ATCC25922 and K. pneumoniae ATCC700603 were used as reference strains for susceptibility testing. Meropenem and cefoperazone/sulbactam (2:1) were determined by the K-B method as recommended by the Clinical Laboratory Standards Institute (CLSI) of 2009 version .The identity and susceptibility of isolates were confirmed using the VITEK system . Enterobacteriaceae were confirmed using an ESBL confirmatory test according to CLSI guidelines are shown in Enterobacteriaceae increased dramatically (about 20-fold) from 2000 to 2009. During the first 5 years of the survey (2000\u20132004), the number of E. coli was higher than that of K. pneumonia. From 2005 onward, K. pneumoniae was the most prevalent species of Enterobacteriaceae, followed by S. marcescens and E. cloacae.The most commonly isolated E. coli, K. pneumoniae, Klebsiella oxytoca, and P. mirabilis. The percentage of E. coli and K. pneumoniae that produced ESBLs demonstrated a gradual and remarkable increase from 7.2% in 2000 to 56.6% in 2007, and from 11.5% in 2000 to 68.5% in 2007, respectively, while this percentage decreased in 2008 and 2009. ESBL-producing P. mirabilis was detected from 2006 onward remained at about 50%, resistance rate to carbapenems increased yearly, but remained under 10%. Resistance rate of S. marcescens to carbapenem suddenly increased to 40\u201360% in 2005 and 2006 due to an outbreak of carbapenem-resistant S. marcescens as reported typing showed that most of these S. marcescens isolates belonged to the same clone. The K. pneumoniae species that were isolated from the Second Affiliated Hospital of Zhejiang University were classified into five dominant clones over this decade. Resistance rate to gentamicin also declined over this decade. However, the percent of E. coli resistant to gentamicin remained at over 50%, which was higher than the resistance rate of K. pneumoniae. A high percentage of E. coli (60%) was also resistant to ciprofloxacin and levofloxacin, and this resistance rate was higher than that of other Enterobacteriaceae isolates, consistent with the 2008 CHINET survey of bacterial resistance in China in North America, Europe, Latin America, Middle East, Africa, and Asia from 2002 to 2007 showed that imipenem and meropenem showed good activity against eriaceae . Similareriaceae , 13, 14. in 2004 . All \u03b2-l in 2001 and was lla spp. \u201318. blaKg, China , althougK. pneumoniae that was detected was higher than that of E. coli, especially in 2009. Moreover, PFGE typing showed that the S. marcescens isolates that were isolated in hospitals in Hangzhou during the epidemic of carbapenem-resistant S. marcescens that occurred in 2005\u20132007 belonged to the same clone, and most of them harbored blaKPC-2 (Enterobacteriaceae to antibiotics containing enzyme inhibitors (such as sulbactam or tazobactam) also increased over the survey period, and this trend toward increased resistance accorded with the increased resistance to carbapenems. For instance, both K. pneumoniae and S. marcescens showed high resistance to carbapenems, and their resistance rate to antibiotics containing enzyme inhibitors was also very high (2005\u20132007).In addition, the percentage of carbapenem-resistant Enterobacteriaceae indicated that more resistant isolates were identified from coastal and developed cities than from rural and mountain areas, and that the resistance rate of the isolates from developed cities was much higher than that from rural areas. Hangzhou , Wenzhou, Ningbo, and Taizhou are coastal cities, while Shaoxing is an industrially developed area. These areas and cities are economically developed with higher income per capita and more frequent use of antibiotics, especially of broad-spectrum antibiotics, than rural areas such as in Lishui and Quzhou. Economically developed areas have a relatively developed medical system with a higher chance of antibiotic exposure that will increase the possibility of bacterial resistance. Moreover, the higher population density in these areas also increases the chance that resistant pathogens will be transferred among the population. While social-economic development has helped to improve the living standard of the people, it has also changed the micro-flora, especially the pathogens, and increased bacterial resistance, which poses a challenge to our future medical care and infection-control strategies. This finding is in line with our previous report that, during the last several decades, the ratio of Shigella spp. (Shigella flexneri vs. Shigella sonnei) in the province has undergone a shift from a ratio that is typical of developing countries to a ratio that is typical of industrialized countries (The geographic distribution of carbapenem-resistant ountries .Enterobacteriaceae, is a serious problem in Zhejiang, and should be further monitored in the coming years. Much restrictive actions have to be taken in terms of rational use of antibiotics and nosocomial control to prevent the further spread of the multi-resistant pathogens.Currently, the emergence of multi-resistant gram-negative bacteria, especially of"} +{"text": "There was an error in the Funding Statement. The Funding Statement should read: \u201cThis work was supported by ENFIN, a Network of Excellence funded by the European Commission within its FP6 Programme, under the thematic area 'Life sciences, genomics and biotechnology for health' (contract number LSHG-CT-2005-518254). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "The eruption of Kaposi sarcoma (KS) and aggressive non-Hodgkin lymphoma (NHL) in young homosexual men in 1981 in the West heralded the onset of the human immunodeficiency virus (HIV) infection epidemic, which remains one of the biggest challenges to global public health and science ever. Because KS and NHL were increased >10,000 and 50-600 times, respectively, with HIV, they were designated AIDS defining cancers (ADC). Cervical cancer (CC), increased 5-10 times was also designated as an ADC. A few other cancers are elevated with HIV, including Hodgkin lymphoma (10 times), anal cancer (15-30 times), and lung cancer (4 times) are designated as non-AIDS defining cancers (NADCs). Since 1996 when combination antiretroviral therapy (cART) became widely available in the West, dramatic decreases in HIV mortality have been observed and substantial decrease in the incidence of ADCs. Coincidentally, the burden of NADCs has increased as people with HIV age with chronic HIV infection. The impact of HIV infection on cancer in sub-Saharan Africa, where two thirds of the epidemic is concentrated, remains poorly understood. The few studies conducted indicate that risks for ADCs are also increased, but quantitatively less so than in the West. The risks for many cancers with established viral associations, including liver and nasopharynx, which are found in Africa, do not appear to be increased. These data are limited because of competing mortality, and cancer is under diagnosed, pathological confirmation is rare, and cancer registration not widely practiced. The expansion of access to life-extending cART in sub-Saharan Africa, through programs such as the Global Fund for AIDS, Malaria, and Tuberculosis and the US President's Emergency Program for AIDS Relief (PEPFAR), is leading to dramatic lengthening of life of HIV patients, which will likely influence the spectrum and burden of cancer in patients with HIV. In this paper, we review current literature and explore merits for integrating cancer research in established HIV programs to obtain timely data about the incidence and burden of cancer in HIV-infected persons in Africa. The global cancer burden has been increasing rapidly over the past 30 years , both inth International Conference of the Institute of Human Virology at Tropea, Italy, in 2010. We summarize the consensus that collaboration with infectious disease HIV programs in sub-Saharan Africa may provide practical opportunities for research, treatment and prevention about cancer in HIV infected populations.Although 70% of the global HIV/AIDS epidemic is concentrated in sub-Saharan Africa ,20, the The bulk of our knowledge about HIV and cancer comes from studies conducted in the West (2448 of 2587 case-referent studies ), althouKS was rare in the United States before the AIDS epidemic. Among white men in San Francisco, an early epicenter of the AIDS epidemic, incidence of KS rose steeply from 0.5 per 100,000 people/year in 1973 to a peak of 33.3 in 1991 , but mirIn contrast to KS, NHL was relatively common in the general population in the U.S. and the incidence was rising before the arrival of the HIV epidemic . ImpressThe risk pattern for CC contrasts that of KS and NHL. Modest risk elevation of 4.2 times during 1990-95 and 5.3 during 1996-2002 was noted . The proThe introduction of cART in the West in 1996 ,35 and iThe incidences of several other cancers, including lung, anus, liver, and Hodgkin lymphoma are increased with HIV/AIDS. These cancers are currently considered NADCs, and the reasons for the increased incidences are varied. For example, lung cancer incidence has consistently been shown to be increased 3-4 times higher in persons with HIV/AIDS ,38,39. AIntriguingly, breast and prostate cancer incidences rates appear decreased with immunosuppression ,42. The The incidence rates of most common epithelial cancers, such as colon cancer, are not increased ,38. AbseAlthough the region is home to about 70% of the AIDS pandemic, only about 139 (5%) studies have been conducted to examine the impact of HIV and cancer in sub-Saharan Africa . One of et al., [KS was endemic in East and Central Africa before the AIDS epidemic accounting for 5-18% of cancers ,10. Duriet al., has advaet al., [In contrast to KS, the risk for NHL in the general population in sub-Saharan Africa is relatively low, except et al., found NHet al., . The medet al., . This inet al. [et al. [The risk of NHL with HIV has been quantified in a few studies Table . In Rwanet al. found an [et al. found an [et al. . The SIR [et al. . However [et al. .+ counts in sub-Saharan Africa than in the West [The reasons for the comparatively lower risk of NHL in sub-Saharan Africa relative to rates observed in the West may be artifactual due to under-diagnosis or may be due to competing mortality or environmental/genetic factors. Survival with HIV may be too short due to competing mortality from common infections, such as malaria and tuberculosis , to permthe West . Howeverthe West is compathe West . Might rthe West , perhapsIs it possible that decreased risk of NHL might be due to prevalent exposure linked to treatment of common infections in the region? For example, antimalarials, including chloroquine, are widely used in Africa through formal prescription as well as self-medication for fever is prevalent . In thiset al., [Cervical cancer is the most common cancer in women in most countries in sub-Saharan Africa ,73, but et al., observedet al., ,73. The et al., . These ret al., [3 compared to those with higher counts [et al., [The prevalence of cervical intraepithelial neoplasia (CIN) or human papilloma virus (HPV) infection is elevated 2-6 times in HIV-positive women than in HIV-negative women in east ,85, westet al., found ceet al., . The linet al., . The risr counts , althoug[et al., , found c[et al., . In thisDespite the small risk increases reported, CC is the most important cancer in HIV infected populations in Africa because women account for >50% of HIV epidemic. The public health impact of CC could be addressed by harnessing the historical interest and increased funding of HIV/AIDS treatment and prevention programs to support new initiatives for CC screening and treatment . This apet al. have summarized the comprehensive literature on the association between HPV and SCCC [Squamous cell carcinoma of the conjunctiva (SCCC) is a rare tumor of the ocular surface, which is linked to ultraviolet radiation exposure and, based on elevated risk with HIV , appearaand SCCC The studand SCCC . The freand SCCC . The stuHodgkin lymphoma, although not designated as AIDS defining, is consistently elevated with HIV in most studies ,104. TheHepatocellular carcinoma (HCC) was relatively common in men in Africa before the AIDS epidemic, in part, because of the high prevalence of hepatitis B virus (HBV) infection and exposure to aflatoxin. There iEvaluation of cancer statistics must focus on the quality of data available to support scientific and public health initiatives. Although based on the best data currently available, the impact of HIV on the incidence and burden of cancer is probably underestimated. Only about one third of people with HIV in sub-Saharan Africa know about their infection and onlyThe study of cancer in HIV persons in Africa is valuable for several reasons. First, the large size of the HIV epidemic underscores its public health significance, including in its impact on cancer. Two, Africa encompasses extraordinary genetic diversity of pathogens and hosts, which hold promise for unique opportunities to learn about the biology of infection, immunology and cancer. Two HIV types are relevant to the epidemic (HIV-1 and HIV2), but HIV-1 is responsible for 95% of HIV infections globally. HIV-1 is divisible into ten subtypes and some circulating recombinant forms are recognized , which hGlobally coordinated efforts aimed at interrupting the spread of, and mortality from, HIV/AIDS in sub-Saharan Africa present opportunities to study cancer in individuals with HIV/AIDS on the continent . Millennhttp://www.iedea-hiv.org), which was launched with funding from the National Institutes of Health, to collect, harmonize, and standardize data from the continent to allow comparative analysis of common and dissimilar impacts [http://www.aortic.org/) has a mission is to promote cancer awareness and improve cancer diagnosis and treatment in Africa. With its network across the continent and bi-ennial scientific meetings, AORTIC provides a resource that could be leveraged to initiate, implement, and report on Africa-wide studies of cancer. The recent focus large biomedical centers, such as the National Cancer Institute at NIH, by establishing Centers for Global Health[The resource-focused approach would aim to leverage large networked or linked clinics ,126,127 impacts . The Afral Health is timelhttp://www.ssalc.org/acsr_drupal/). These investigators will bring clarity to the question of the spectrum of lymphomas diagnosed with HIV in Africa and their diagnostic support is likely to spur epidemiological and clinical studies of NHL. We noted outstanding questions about the etiology of SCCC [et al. and co workers [In contrast to the resource-focused approach, in the disease-focused approach, specific hypotheses are raised and specific studies designed to answer those questions. For example, the question of the types and risk of NHL is currently being addressed by a consortium of investigators focused on lymphomas ( of SCCC ,99. Dise of SCCC . Hessol workers have lin workers ,132 and workers . Thus, c workers . HomosexThe authors declare that they have no competing interests.SMM drafted the manuscript. All authors reviewed and critically revised the manuscript and approved the final draft."} +{"text": "Dome-type carcinoma (DC) is a distinct variant of colorectal adenocarcinoma and less than 10 cases have been described in the literature. Most of the previously reported cases were early lesions and no endoscopic observations have been described so far. We herein report a case of a DC invading the subserosal layer, including endoscopic findings.A highly elevated lesion in the transverse colon was diagnosed by colonoscopy in a 77-year-old man. The tumor appeared to be similar to a submucosal tumor (SMT), however, a demarcated area of reddish and irregular mucosa was observed at the top of the tumor. There were no erosions or ulcers. Laparoscopic-assisted right hemicolectomy was performed and pathological examination revealed a well-circumscribed tumor invading the subserosal layer. The tumor was a well-differentiated adenocarcinoma associated with a dense lymphocytic infiltration and showed expansive growth. The overlying mucosal layer showed high-grade dysplasia.The present lesion was diagnosed as a DC of the colon invading the subserosal layer. Because the association of mucosal dysplasia is common in DCs, the detection of dysplastic epithelium would be important to discriminate DCs from SMTs. Since Jet al. ,2 reportet al. ,4. Basedet al. .We herein report a case, along with the endoscopic findings, of a DC invading the subserosal layer.A 77-year-old man suffered abdominal discomfort and underwent a total colonoscopy. The colonoscopy identified a highly elevated lesion, 30 mm in diameter, in the transverse colon Figure . The tumPathological examination revealed a well-circumscribed tumor invading the subserosal layer Figure . The tumet al. [Jass et al. ,2 reportet al. -8.Generally, prominent lymphocytic infiltration is known as a feature of colorectal cancers with a microsatellite instability-high phenotype and tumors with EBV infection. However, the present case, and the majority of the previously reported DCs, did not show evidence for microsatellite instability, as examined by either microsatellite instability test or immunohistochemistry for mismatch repair proteins, and EBV infection . The lacAll but one previously reported DCs were early cancers limited to the submucosal layer . It has Endoscopically, the present case resembled SMT, reflecting the expansive growth of the tumor. However, while the base of the lesion was covered with non-neoplastic mucosa, an area of mucosal dysplasia could be endoscopically detected on the top of the lesion, and a biopsy taken from this area allowed a diagnosis of adenocarcinoma. Because the previously reported DCs also lacked erosion or ulceration and were associated with mucosal dysplasia -4,7, theEven though the current classifications do not recognize DC as a distinct histological subtype, the present and previous reports illustrated peculiar histological and clinical characteristics of DC. Further accumulation of cases and phenotypical characterization, including the potential relationship to M-cells, may establish DC as a distinct subtype of colorectal adenocarcinoma.Written informed consent was obtained from the patient for publication of this case report and any accompanying images. A copy of the written consent is available for review by the editor-in-chief of this journal.DC: Dome-type carcinoma; SMT: Submucosal tumor; EBV: Epstein-Barr virus.The authors declare that they have no competing interests.MY for design and drafting of the manuscript; Dr. SS for the concept and the revision of the manuscript and the pathological diagnosis; Dr. TM for the revision of the manuscript and the supervision; Drs. MY, HT, and RK for the pathological diagnosis; Drs. TS, TN and YS for the endoscopic diagnosis; Dr. TA for the surgical treatment. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-230X/12/21/prepub"} +{"text": "There was an error in the Funding statement. The correct Funding is:Financial support was provided by the Spanish MEC (BFU2009-07286), Spanish MICINN (EUI2009-04083) in the frame of the ERA-NET NEURON, and JCYL-UE (GR221) to Dr. Malmierca and the NSF (IOS-0719295) to Dr. Covey. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "There is a grant number missing from the Funding Statement. The following is the complete Funding Statement:This work was funded by national funding of FCT (Portuguese Foundation for Science and Technology) and by FEDER through COMPETE under de project Micro2Micro - PTDC/EBB-EBI/104263/2008. C. Almeida also acknowledge FCT for the PhD Fellowship - SFRH/BD/29297/2006. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Distance learning may be useful for building health research capacity. However, evidence that it can improve knowledge and skills in health research, particularly in resource-poor settings, is limited. We compared the impact and acceptability of teaching two distinct content areas, Biostatistics and Research Ethics, through either on-line distance learning format or traditional on-site training, in a randomized study in India. Our objective was to determine whether on-line courses in Biostatistics and Research Ethics could achieve similar improvements in knowledge, as traditional on-site, classroom-based courses.Subjects: Volunteer Indian scientists were randomly assigned to one of two arms.Intervention: Students in Arm 1 attended a 3.5-day on-site course in Biostatistics and completed a 3.5-week on-line course in Research Ethics. Students in Arm 2 attended a 3.5-week on-line course in Biostatistics and 3.5-day on-site course in Research Ethics. For the two course formats, learning objectives, course contents and knowledge tests were identical.Main Outcome Measures: Improvement in knowledge immediately and 3-months after course completion, compared to baseline.Baseline characteristics were similar in both arms (n = 29 each). Median knowledge score for Biostatistics increased from a baseline of 49% to 64% (p < 0.001) 3 months after the on-site course, and from 48% to 63% (p = 0.009) after the on-line course. For the on-site Research Ethics course, median score increased from 69% to 83% (p = 0.005), and for the on-line Research Ethics course from 62% to 80% (p < 0.001). Three months after the course, median gains in knowledge scores remained similar for the on-site and on-line platforms for both Biostatistics and Research Ethics .On-line and on-site training formats led to marked and similar improvements of knowledge in Biostatistics and Research Ethics. This, combined with logistical and cost advantages of on-line training, may make on-line courses particularly useful for expanding health research capacity in resource-limited settings. There is great need and demand for building health research capacity globally, to ensure that opportunities and competence to undertake scientifically and ethically sound research exist in all regions ,2. CurreDistance learning, including on-line teaching, may provide an attractive, cost-effective, scalable and efficient alternative to on-site classroom training. On-line education formats use diverse media such as text, images, audio, video and interactive formats . On-lineDistance education has also supported training of students in resource-limited settings. India has the largest distance learning university in the world, offering nearly 350 courses with approximately 2.8 million students currently enrolled ,13. HoweTo better understand the potential value of on-line learning platforms to expand health research capacity, we undertook a randomized study comparing on-line with on-site (i.e. face-to-face) delivery of information in two distinct domains relevant for international health research: Biostatistics and Research Ethics. Further, to assess the feasibility and potential of utilizing on-line platforms for expanding health research capacity in resource-limited settings, we conducted this study in India. Our hypothesis was that both on-site and on-line course formats would lead to similar gains in knowledge for students, for both content domains. We further hypothesized that trainees would report a similar level of satisfaction for on-line and on-site course platforms.Volunteers for the study were recruited through an announcement in Indian biomedical journals and via email invitations to individuals engaged in biomedical research, and leaders of medical schools and major institutions conducting health research in India. The eligibility criteria for inclusion were: (i) a degree in medicine or a masters' degree in science, (ii) receipt of a graduate or postgraduate degree within the last ten years, (iii) at least one-year of experience in human health-related clinical or social science research, (iv) basic computer skills and availability of broadband internet access, (v) willingness to be randomized and to participate in the study, and (vi) willingness to undertake pre- and post-course evaluations. Interested persons were invited to register at a website, and respond to questions relevant to eligibility. Four investigators reviewed the applications.The study used a randomized design. Following informed consent, each study participant was allocated to one of two study arms, using a computer-based randomization procedure. Participants in Arm 1 traveled to Lucknow, India for a 3.5-day classroom training in Biostatistics, and a week later participated from their own home or office settings in a 3.5-week on-line training course in Research Ethics . The Biostatistics course included 17 lectures over 15.5 hours, that covered statistical analysis and study design, including types of data, descriptive statistics, normal distribution, sampling distribution, central limit theorem, confidence intervals, hypothesis testing for continuous and categorical data, simple linear regression, measures of association, survival analysis, and design of observational and interventional studies. Students also participated in 8 interactive group exercise sessions of 45-60 minutes each designed to apply knowledge gained during the lectures. On-line students were provided the same exercises to discuss and solve on-line with other students and the course faculty, during 8 interactive sessions. All lectures for the on-line format were pre-recorded by JM. He was unable to travel to India for the on-site course, and the three Indian faculty experts in biostatistics delivered lectures, using the same slides as those used in the on-line course. They interacted via email with JM and repeatedly listened to his on-line lectures, in order to deliver the on-site live lectures consistent with JM's on-line lectures. The 8 interactive sessions for both the on-line and on-site formats were conducted by two Indian faculty experts .The Research Ethics course was developed by faculty experts at the Johns Hopkins Berman Institute of Bioethics (NK and HT), with input on learning objectives, course content and knowledge assessments contributed by a faculty expert from India (AB). The three Research Ethics faculty delivered 15 lectures with 8.75 hours of instruction, covering ethical principles, a framework for ethical analysis, informed consent, the relationship of study design to ethics, risk/benefit assessment, the role of ethics committees, privacy/confidentiality, and honesty in science. In addition, students participated in 5 one-hour interactive case discussions, requiring application of ethical analysis skills. Students also viewed and discussed one 20-minute video on ethical challenges in community-based research. Course participants completed two homework assignments. The same faculty experts from the US and India provided the lectures and cases discussions for both the on-line and on-site formats of the Research Ethics courses.The interactive sessions for online courses were conducted at pre-specified times and lasted about 60 minutes each. Each session was moderated by one or more course faculty. The course participants were encouraged to log in into these sessions over the Internet, though attendance was not compulsory. During these online 'classrooms', faculty could use audio to address student questions asked via a typed message that appeared on the faculty members computer screen as well as the screen of all other students. The faculty could also share his/her computer screen, on which s/he could write and draw. Further, the moderator could give 'audio' rights to any student, allowing him/her to speak to the entire class.For the statistics online course, eight online sessions were so arranged that each topic was covered in two consecutive sessions to allow a participant who could not join a particular session to 'attend' the other; each participant was therefore expected to attend four sessions. The participants were encouraged to send to moderators any questions that they wanted discussed. The faculty member reviewed the question and answers for these with the students.The research ethics faculty held five case-based discussions with the students in the on-line course. Three sessions were held for each case-based discussion, each moderated by a member of the faculty. Students were given the case and a set of questions to answer in advance of the session. The faculty member would review the question and answers with the students and then moderate a discussion on the case.For each training course , all participants were administered knowledge tests before the course, as well as immediately and three months after course completion. These tests focused on assessment of participants' knowledge related to the specific learning objectives and application of this knowledge to problems raised in case histories. Responses were scored against pre-determined answers, and an overall knowledge score (percent correct responses) was computed for each test. Gains in knowledge scores in each domain (Biostatistics or Research Ethics) were compared between the participants receiving on-site and on-line training. The tests of knowledge were the same for on-line and the on-site platforms in each content domain. The biostatistics knowledge tests consisted of 20 questions in objective format . The research ethics knowledge assessments given just before and after the course were unique sets of 41 multiple choice, true/false and short answer questions. The three month follow-up knowledge assessment consisted of the 41 best performing questions from the pre and immediate post-course assessments.The gain in knowledge at 3 months after completion of each course was used as the primary outcome measure. In addition, at the completion of each course, each study participant was administered a course evaluation questionnaire to assess student acceptability and satisfaction with the course. This assessment utilized a 5-point Likert scale for most items; for a few questions, 4-point and 3-point Likert scales were used.The study was approved by ethics committees at both the participating institutions . Each study participant signed a written informed consent prior to randomization, and submitted it by fax, as scanned computer file by email or through postal mail.Wilcoxon's rank sum test and Wilcoxon's signed rank test were used for inter-group and paired comparisons, respectively. Multivariate quartile regression analyses were used to determine independent predictors of gain in knowledge scores at 3 months following course completion, in each domain. Covariates, including age, number of years since last degree, and mode of training (on-site versus online), were tested in univariate analyses as predictors of gain in knowledge scores in biostatistics and in research ethics separately. Those found significant at p < 0.05 level were included in a multivariable analysis. Data on acceptability of and satisfaction with each course were compared using a trend test for ordinal data. In addition, the effect of demographic factors on the relationship between gain in knowledge at 3 months from baseline and the training platform (on-line or on-site) was examined using a quartile regression analysis; factors found significant on univariate analysis were entered into a multivariate analysis. Data on acceptability of and satisfaction with each course and mode of instruction were compared using a chi-squared test for trend for ordinal data.A total of 250 persons registered on-line for participation in the study, and 75 volunteers were invited to submit a completed and signed consent form. Of these, 60 invited volunteers agreed to participate, submitted signed consent forms and were randomized to either Arm 1 or Arm 2. Two volunteers, one randomized to each arm, subsequently elected to drop out of the study prior to initiation, due to inability to travel on the dates of the on-site course or to attend the on-line course, respectively. Thus, a total of 58 volunteers participated. Their median age was 34 years [Range 25-48 years] and 45 (78%) were male. Participants in each arm were similar in age, gender distribution, number of years since obtaining last degree, and pre-training baseline knowledge scores in both Biostatistics and Research Ethics domains and on-line course (p = 0.009) formats Table . When asResearch Ethics knowledge scores also increased significantly from baseline, for both the on-site and on-line platforms , than those in the on-line course format was significantly associated with knowledge gain on univariate analysis (data not shown). However, knowledge gain in Research Ethics was significantly associated with younger age and lower baseline knowledge scores on univariate analysis. On multivariate analysis, only lower pre-course knowledge score was independently associated with a greater gain in Research Ethics knowledge . For both univariate and multivariate analyses, instructional platform (on-line vs on-site) was not associated with significant differences in knowledge gain for Research Ethics (data not shown).Overall, a very high level of student satisfaction (\"Agree\" or \"Strongly Agree\") was demonstrated for both on-site and on-line formats, and for both Biostatistics Tables , 4 and 5Our study comparing on-line to on-site teaching formats in a resource-limited setting demonstrated that both formats significantly increased knowledge from baseline and this increase in knowledge was observed for both content areas (Biostatistics and Research Ethics). In addition, the increases in knowledge were sustained for 3 months after completion of the courses. There was a high level of student satisfaction for all courses, although the on-site format was associated with a somewhat higher level of satisfaction related to instructor accessibility and quality of faculty feedback.A recent meta-analysis of 126 on-line learning interventions demonstrated that all but two of these were associated with a gain in knowledge . Most ofIn another meta-analysis covering the impact of on-line training in diverse fields from school level to professional level , on-lineper se. Alternatively, the difference may be related to better learning during on-site training, possibly due to the influence of factors such as face-to-face rather than screen-to-screen interaction with the instructor.Although results were similar when assessed 3 months after the courses, for both knowledge domains, we found higher knowledge scores, particularly for Biostatistics, immediately after the on-site courses, than after the on-line course. Several factors may account for this observation. First, our study participants may not have had internet access of reasonable quality. Although we verified each participant's access to sufficient broadband internet connectivity at the beginning of the study, intermittent outages may have occurred. Second, participants in a resource-limited setting may not be as familiar with on-line courses as in US universities where 20-25% of all students take at least one on-line course , the betThe difference in immediate knowledge gain between the on-line and on-site formats was more marked for Biostatistics, than for Research Ethics. A possible explanation of this difference is that the quantitative skills required for Biostatistics may be poor in medical professionals. Greater faculty-trainee interaction during on-site training may thus be helpful when training medical professionals in this domain. The results of the participant satisfaction survey, in which the on-site course scored higher in questions related to faculty-trainee interaction, would support this explanation. The greater satisfaction with on-site courses may also mean that further efforts are needed to enhance faculty-trainee interaction during on-line training.Our study has several potential limitations. First, participants in our study may not be adequately representative of the population of prospective biomedical researchers that require training in research methodology. Our study participants may have been highly motivated, and therefore likely to have greater gains in knowledge scores with any type of training. However, due to the randomized design, this effect would have been similar for on-line and on-site courses, and would not affect our conclusions of comparative performance of the two training formats. Another potential limitation is that our study participants were from India and our results may not be generalizable to other populations with less access to and familiarity with the internet. Finally, although both the on-line and on-site Research Ethics courses used the same faculty, this was not possible for the two Biostatistics courses. However, in a previous meta-analysis, it was shown that a change in faculty member did not influence effectiveness of an on-line course, provided the course content and method of delivery were unchanged .Another limitation of our study, designed as a comparative efficacy trial of two formats of training, was the inability to accurately assess the costs per person for each training format. Attendance at a course entails several different types of costs, including course fees, costs of travel and accommodation, and costs due to lost wages or work, etc; further, the course fees include costs of faculty time, course material, and facilities used. Ours being a research study, travel and hotel accommodation for all participants were arranged on a uniform scale, and these had no relationship with the costs that the participants would have incurred if they had arranged and paid for these. Because we used some of the pre-existing course materials and online facilities, costs of development of new courses for either on-site or online training courses could not be assessed accurately. Also, for the on-site ethics course, faculty members travelled from USA to India; this expenditure is unlikely to occur in real-life. Further, the 'cross-over' study design limited the number of participants in the online courses, precluding true assessment of per capita costs. Thus, in view of the unusual settings, we could not compare comparison of costs incurred per capita for the two training formats.A training program can be evaluated at various levels. A popular approach to evaluation of training, the Kirkpatrick evaluation model, delineates four levels of learning outcomes ,17. ThesOn-line teaching provides several advantages over more traditional on-site course formats, particularly for building research capacity in resource-limited settings. On-line courses can be more flexible, convenient and accessible ,18, partIn our randomized controlled study, On-line and on-site training formats led to marked and similar improvements of knowledge in Biostatistics and Research Ethics. Since on-line training has several logistical and cost advantages over on-site training, distance learning using on-line tools may be a particularly useful, efficient, cost-effective and scalable strategy for expanding health research capacity in resource-limited settings.US: United States; USA: United States of America;The authors declare that they have no competing interests.RA, NG, NK, HT, AB, AA, SDS, JM, PM and RCB were involved in conception and design of the study. RA, NG, HT, JA, AB, AA, SDS, SK, JM, JMW, RCB played a role in acquisition of data. RA, NG, HT, JA, AB, AA, RCB were involved in analysis and interpretation of data. The initial manuscript draft was prepared by RA, AA, JMW and RCB. All the authors participated in critical revision of manuscript for intellectual content. NG did the statistical analysis. RA and RCB played a role in obtaining funding for the study. RA, NG, JA, AB, AA, SK, JMW and RCB provided administrative, technical or material support. RA, NG, AA, SK, PM and RCB supervised the study. All the authors have read and approved the final manuscript. RA and RCB had full access to all of the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis. The sponsor played no role in data analysis or in the decision to publish this paper.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1472-6920/11/37/prepub"} +{"text": "We reported that the compositions of arachidonic acid (ARA) in erythrocytes and plasma phospholipids (PL) in the elderly were lower than those in the young, though the ARA intake was nearly identical.We further analyzed data in four study groups with different ages and sexes, and determined that the blood ARA levels were affected by the kinds of dietary fatty acids ingested.One hundred and four healthy young and elderly volunteers were recruited. Dietary records together with photographic records from 28 consecutive days were reviewed and the fatty acid composition in plasma lipid fractions and erythrocyte PL was analyzed.No correlations for ARA between dietary fatty acids and blood lipid fractions were observed. A significant negative correlation between eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) intake and ARA composition in erythrocyte PL was observed. ARA composition in erythrocyte PL was significantly lower in elderly subjects than in young subjects, because EPA and DHA intake in elderly subjects was higher than in young subjects. However, after removing the effect of dietary EPA+DHA intake, the ARA composition in erythrocyte PL in elderly subjects was significantly lower than that in young subjects.Changes in physical conditions with aging influenced the low ARA composition of erythrocyte in elderly subjects in addition to the effects of dietary EPA and DHA. Arachidonic acid (ARA), one of the n-6 polyunsaturated fatty acids (PUFA), is the predominant fatty acid (FA) of membrane phospholipids (PL) in mammalian brain and neural tissues ,2. ARA rMany studies in the last decade have shown the role of sufficient intake of n-3 PUFA in the prevention of several diseases, particularly coronary heart disease -7. Eicoset al. [Hereafter, we think that our attention will be focused on blood ARA for the associations with disorders; therefore, we should understand how the blood ARA is changed by factors including the discrepancies in diet, sex, and age. Weseler et al. reportedet al. . Thus, tet al. -18.We conducted a dietary survey using dietary records together with photographic records over 28 consecutive days and determined the FA compositions in plasma triacylglycerol (TG), esterified cholesterol (EC), and PL, and erythrocyte membrane PL in four study groups: young men, young women, elderly men, and elderly women. We reported that the compositions of ARA in plasma and erythrocyte PL in the elderly were lower than those in the young, though the ARA intake was nearly identical . In thisThe details of the subjects are reported in our previous study . BrieflyA dietary record was continuously maintained using a written form together with photographic records for 28 consecutive days. Although every investigation was conducted in early summer, the investigation year was 2004 for the young women, 2005 for the elderly men, 2007 for the elderly women, and 2008 for the young men. The details of the dietary survey method are reported in our previous study in JapanFasting blood sampling was conducted on the day after completion of the 28-day dietary survey. After blood sampling, the samples were centrifuged to separate plasma and erythrocytes. The preparation of erythrocyte membranes and the analysis of FA compositions were conducted as described previously . BrieflyThe relationships between dietary FA and FA in plasma TG, EC, and PL and erythrocyte membrane PL were examined by Spearman's rank correlation. Statistical differences among the four groups were determined using ANOVA and the Bonferroni post hoc test. We calculated the least square means and 95% confidence intervals of ARA composition in erythrocyte PL using analysis of covariance (ANCOVA) models, with EPA+DHA intakes as the covariables and ARA composition in erythrocyte PL as the dependent variable, because it was possible that dietary EPA+DHA intake affected the ARA composition in erythrocyte PL.P < 0.05. We conducted calculations using the Statistical Package for Social Science software .A significant difference in analysis results was observed at Spearman's correlation coefficients between dietary EPA, DHA and ARA intake and the corresponding FA composition of plasma TG, EC, and PL and erythrocyte PL in YM-G, YW-G, EM-G, EW-G, and the total subjects group (ALL-G) are shown Table Spearman's correlation coefficients between dietary EPA and DHA intake and ARA composition of plasma TG, EC, and PL and erythrocyte PL in YM-G, YW-G, EM-G, EW-G, and ALL-G are shown in Table A scatter plot between dietary EPA+DHA intake and ARA compositions in erythrocyte PL is shown in Figure P < 0.001), the slope of the regression line was -2.3, and the F value of non-parallelism was 0.54 (P = 0.65). As a result, the effect of EPA+DHA intake on ARA compositions in erythrocyte PL was not negligible and the regression lines in YM, YW, EM and EW were parallel. We then calculated the adjusted mean and standard error of the mean (SEM) of ARA composition in erythrocyte PL in YM-G, YW-G, EM-G, and EW-G; these results are shown in Table P < 0.001).Because dietary EPA+DHA intake seems to affect the ARA composition in erythrocyte PL, we analyzed by ANCOVA with EPA+DHA intakes as the covariables. As a result, the F value of common regression was 40.8 dietary ARA intakes were not correlated with the composition of ARA in erythrocyte PL, but dietary EPA and/or DHA intakes were negatively correlated with the composition of ARA in erythrocyte PL in all subjects groups and (2) after removing the effect of dietary EPA+DHA intake, the ARA composition in erythrocyte PL was significantly lower in elderly subjects than in young subjects.et al. [et al. [In previous survey among Japanese people, a negative correlation between ARA intake and serum PL level of ARA was observed . Converset al. reportedet al. ,17. Simi [et al. showed net al. [Yanagisawa et al. reportedet al. . Our datet al. . We indiErythrocyte membrane FA composition is affected by diet and is considered to reach a new steady state level 4 to 5 weeks after the establishment of new diet management . We perfIn summary, the ARA levels in blood all lipid fractions, especially in erythrocyte PL, were affected by the amount of EPA and/or DHA intakes. The ARA composition in erythrocyte PL was significantly lower in elderly subjects than in young subjects, because EPA and DHA intakes in elderly subjects were higher than in young subjects. However, after removing the effect of dietary EPA+DHA intake, the adjusted ARA composition of erythrocyte PL in elderly subjects was significantly lower than that in young subjects. Consequently, changes in physical conditions with aging influenced the low ARA composition of erythrocyte in elderly subjects in addition to the effects of dietary EPA and DHA.The authors declare that they have no competing interests.TK conceived the study, participated in the design of the study, acquired data, performed the statistical analysis and drafted the manuscript. SH, TH, NA, YK, NI and KK carried out the survey, measured blood fatty acid compositions, and organized the data. EA, HK and YK participated in the design of the study and helped conducting the study. All authors read and approved the final manuscript."} +{"text": "Published population-based studies have used variable methodology, which has influenced findings and made comparisons difficult. The Global Campaign against Headache is undertaking initiatives to improve and standardize methods in use for cross-sectional studies. One requirement is for a survey instrument with proven cross-cultural validity. This report describes the development of such an instrument. Two of the authors developed the initial version, which was used with adaptations in population-based studies in China, Ethiopia, India, Nepal, Pakistan, Russia, Saudi Arabia, Zambia and 10 countries in the European Union. The resultant evolution of this instrument was reviewed by an expert consensus group drawn from all world regions. The final output was the Headache-Attributed Restriction, Disability, Social Handicap and Impaired Participation (HARDSHIP) questionnaire, designed for application by trained lay interviewers. HARDSHIP is a modular instrument incorporating demographic enquiry, diagnostic questions based on ICHD-3 beta criteria, and enquiries into each of the following as components of headache-attributed burden: symptom burden; health-care utilization; disability and productive time losses; impact on education, career and earnings; perception of control; interictal burden; overall individual burden; effects on relationships and family dynamics; effects on others, including household partner and children; quality of life; wellbeing; obesity as a comorbidity. HARDSHIP already has demonstrated validity and acceptability in multiple languages and cultures. Modules may be included or not, and others ( Atlas of Headache Disorders and Resources in the World 2011, published by the World Health Organization (WHO) [The global burden of headache is very large -4. The Gon (WHO) :\u201cThe facts and figures [on headache] illuminate worldwide neglect of major causes of public ill-health, and the inadequacies of responses to them in countries throughout the world.\u201dLifting The Burden (LTB), a UK-registered nongovernmental organization, in official relations with WHO. The Campaign\u2019s objectives require action supported by awareness, the latter underpinned by knowledge. The knowledge base is evidence of the levels, nationally and worldwide, of headache-related ill-health and health-care need; it shows what manner of change \u2013 and how much \u2013 is required; it supports the humanitarian, economic and political arguments for change; and it signals the priority that should be accorded to action for change. The knowledge base is the foundation on which everything must be built; it needs to be complete, and sound. Unfortunately it is not.Reduction of the burden of headache worldwide is the central purpose of the Global Campaign against Headache ,2,7, conThe known epidemiology of headache disorders was collated in 2007 , the proThe Global Campaign began to address these deficiencies several years ago, planning population-based studies in Georgia , India , China , Russia Lifting The Burden and its collaborators developed both a standardized protocol and a survey instrument, and tested them empirically, the latter in multiple languages. Here we describe the genesis of the survey instrument, and its evolution into the Headache-Attributed Restriction, Disability, Social Handicap and Impaired Participation (HARDSHIP) questionnaire. Validation studies are reported elsewhere [In the course of planning these studies, lsewhere -13.The process was led by TJS and LJS who, with help from Dr Tarun Dua, Department of Mental Health and Substance Abuse, World Health Organization, conceived the first draft, suggested the areas of enquiry, proposed the question structure and phrasing and established a design template. As population-based studies were planned and undertaken, and in consultation with a wide range of experts and local investigators from over 20 countries, the instrument was amended, expanded through a process of item development and refined through item rejection based on empirical experience. Some questions were rephrased. The studies provided a continuing learning experience, fostering improvements through a series of plan-do-study-act cycles, and were a highly influential part of the development process.During this process, the diagnostic questions were the subject of several validation studies, now completed in India .The HARDSHIP questionnaire cover the following aspects of headache-attributed burden: symptom burden; health-care utilization; disability and productive time losses; impact on education, career and earnings; perception of control; interictal burden; overall individual burden (as willingness to pay for treatment); effects on relationships, love life and family dynamics; effects on others, including household partner and children; quality of life; wellbeing; obesity as a comorbidity.Demographic enquiry is essential to characterize the sample. Data are needed in order to compare those who have been selected with the population of interest from whom they are drawn and of whom they are intended to be representative. While, ideally, these data will reflect all factors that may influence prevalence and/or burden of headache, this objective is necessarily limited by the availability of data characterizing the entire population. National statistics are commonly available for gender and age distributions. Even when they are not, these are of such prime importance in headache epidemiology that they must be known in the sample. Social situation , habitation and ethnicity and/or culture may be important influencers of prevalence or burden, and are therefore of some interest.Diagnosis must follow ICHD criteria because untreated attacks, which they may never have or last had long ago. This results in a high proportion of probable diagnoses because duration criteria appear unfulfilled [Certain criteria distinguishing between migraine and TTH pose particular problems in population surveys . First, ulfilled . Second,ulfilled . False-pMOH is diagnosable in cross-sectional studies only as an association of medication overuse with frequent headache (there is no evidence available of causation) . TherefoSymptoms of common headache disorders include pain, and, of migraine, nausea, vomiting and photo- and/or phonophobia. Symptom burden is addressed in HARDSHIP by questions 14, 15, 20, 21/23, 24, 29\u201332, 36 and 37. Pain can be quantified at individual level as a product of intensity, frequency and duration, and at population level as the product of the average among individuals and prevalence. Nausea, photophobia and phonophobia are almost impossible to quantify, but their occurrence can be recorded and frequencies expressed.Disability attributed to headache is also difficult to quantify completely. Common proxies are lost time and reduced productivity, for which well-validated instruments exist ,20. HARDEnquiry into headache yesterday (effectively HALT-1) (HARDSHIP questions 34\u201345) avoids recall problems almost altogether ,22. It cInterictal burden (HARDSHIP questions 64\u201366) arises because headache attacks are unpleasant, and those who experience them frequently are likely to worry about when the next may occur, and/or attempt to eliminate possible triggers through lifestyle compromise. Interictal burden, which is continuous, is likely to affect subjective wellbeing and may be sufficient to impair quality of life. It is perhaps adequately, if not specifically, captured by measures of subjective wellbeing and quality-of-life measures. HARDSHIP imports, as modules, WHOQoL-8 (questioCumulative burden (HARDSHIP questions 51\u201357), accruing over a lifetime, cannot be fully assessed until late in a lifetime. Furthermore, attribution may be uncertain. Nevertheless, a consequence of recurring inability to work may be decreased probability of promotion, and a consequence of lost school-time may be reduced career opportunities. These may be heavy burdens.say they will pay and what they actually will pay when confronted by the reality, and of course WTP is constrained by ability to pay. Nevertheless, this form of enquiry has been used to assess sustainability of health-care initiatives in resource-poor countries [An overall summary measure of individual burden is unlikely to be comprehensive, but the concept is attractive for its simplicity . One sucountries .Burden on others, unaffected by headache themselves, is addressed by HARDSHIP questions 75\u201386. Subjective interpretations are unavoidable. A full account necessitates enquiries among the others, which in practical terms may be possible only among close family members.Health-care resource consumption (HARDSHIP questions 45\u201350) is relatively easy to enquire into, but subject to recall bias. It should also be easy to establish who pays for it . It is less easy to attach accurate costs to individual items of health care, and this may necessitate separate research into health-care costs in the country or region in question .By far the greater part of the financial cost of headache is the indirect cost of absenteeism and reduced effectiveness at work ,26 , since obesity may be an important and potentially remediable risk factor for frequent headache . Other , and that, although both of these explanations may contribute in part, migraine is indeed more prevalent than past estimates have suggested. Alstadhaug et al. [ie, excluding cases of aura only) was 26.3% (95% CI: 18.5-34.2%), 15.9% in males and 36.7% in females. It is unlikely that Norwegian neurologists are biologically unique.In the studies conducted using HARDSHIP, migraine prevalence has been high, although not in China , where g et al. reportedeg, on comorbidities) added, according to the purpose(s) of the study.For better and comparable population-based studies of the burden of headache, there is a clear need for a survey instrument with proven cross-cultural validity, adaptable to the circumstances of particular studies and resource-availability . HARDSHILifting The Burden; MIDAS: Migraine disability assessment; MOH: Medication-overuse headache; TTH: Tension-type headache; WHO: World Health Organization; WTP: Willingness to pay.CI: Confidence interval; GBD2010: Global burden of disease survey 2010; HALT: Headache-attributed lost time; HARDSHIP: Headache-attributed restriction, disability, social handicap and impaired participation; ICHD: International classification of headache disorders; LTB: Lifting The Burden. GLB, TJS and LJS served as experts in the Global Burden of Disease Survey 2010. GLB has received research funds from the US National Institutes of Health and the Dana Foundation.TJS, GLB, RJ, ZK and LJS are the directors and trustees of TJS and LJS created the original concept and assembled the group of experts. All authors were members of the expert consensus group or engaged in developing the questionnaire in the context of population-based studies, or both. TJS drafted the manuscript. All authors reviewed the manuscript in various drafts and approved the final version.The HARDSHIP questionnaire.Click here for fileDiagnostic algorithm.Click here for file"} +{"text": "A good surveillance is essential to gather information for measures to prevent and control of healthcare-associated infections(HAI). The study is aiming at characterize the HAI surveillance systems(HAI-SS) in 7 States in the South and Southeast region, of Brazil.Cross sectional and descriptive study carried out in two steps: characterization of healthcare system structure by means of consulting of the National Data Base of Healthcare Facilities; interview with person in charge of HAI program. HAI-SS were classified in chain, circle or wheel.A large variation of healthcare facilities were identified. S\u00e3o Paulo is the State with highest number of healthcare facilities but the State of Santa Catarina has the highest ratio of healthcare facilities by 1.000.000 habitants. Human resources for HAI-SS varied, and, in some, they were not exclusive for data management but accumulated other functions. Hospital participation in the HAI-SS was mandatory by law in 3 States. HAI-SS were classified as chain in two, circle in four, and wheel in one State. HAI-SS were mainly driven toward acute care facilities; ventilator associated pneumonia, blood stream, urinary tract, and surgical site infections were included. Participation in the National HAI-SS occurred by sending data regarding blood stream infections. Routine feedback of surveillance data was not adopted in two States, one State have been using data gathered from HAI-SS to develop governmental plans for HAI rates reduction.It was identified inequalities in the healthcare services, potentially inducing to over crowding and posing to risk of low quality in some States. Human resources are insufficient in some States to carry out an adequate governmental plan for reduction of HAI rates. The operational dissimilarities among States may need to be overcome in order to build a good National HAI-SS.None declared"} +{"text": "To propose the 95 patients and 60 controls were enrolled from the University Psychiatry Unit of Catania and from general practitioners (GPs). The patients were divided into four pathological groups: Anxiety, Depression, Anxious-Depressive Disorder and Eating Disorders [Diagnostic and Statistical Manual of Mental Disorders Fourth Edition Text Revision (DSM-IV-TR) official/appendix criteria]. The levels of the cholesterol, triglycerides and apolipoproteins A and B were determined. The LI, for each subject, was obtained through a mathematical operation on the values of the cholesterol and triglycerides levels compared with the maximum cut-off of the general population. The autonomic functioning was tested with Ewing battery tests. Particularly, the correlation between heart rate variability (HRV) and lipid metabolism has been investigated.Pathological and control groups, compared among each other, presented some peculiarities in the lipid metabolism and the autonomic dysfunction scores. In addition, a statistically significant correlation has been found between HRV and lipid metabolism.Lipid metabolism and autonomic functioning seem to be related to the discussed psychiatric disorders. LI, in addition, could represent a new possible biomarker to be considered. The dysfunctions of lipid metabolism and autonomic nervous system have been found to be linked with anxious-depressive spectrum and eating disorders -19, but The aim of this work is to correlate the levels of cholesterol, triglycerides, apolipoproteins A-B, LI and autonomic dysfunction to depression, anxiety, anxious-depressive disorder and eating disorders. In addition, the correlation between HRV and lipid metabolism has been tested.95 patients and 60 controls were enrolled from the University Psychiatry Unit of Catania and from GPs. The demographic characteristics of the pathological and control groups are reported in Table The local ethics committee approved this study , and all participants signed an informed consent.Subjects with familial dyslipidemia, heart disease and psychotic disorders were excluded from the study.Four pathological groups were created: Anxiety, Depression, Anxious-Depressive Disorder and Eating Disorders (ED) . The diagnoses were formulated according to the criteria of the DSM-IV-TR .Figure The autonomic nervous system (ANS) functioning was evaluated with the Ewing battery of tests through an electrocardiograph and a sphygmomanometer; particularly: Deep Breathing, Lying-to-Standing, Valsalva Manoeuvre and Sustained Handgrip tests ,22. ScorParticularly, the correlation between HRV (time-domain method) and lipid metabolism has been investigated.Cholesterol, triglycerides and apolipoproteins A and B were analyzed through fasting blood samples. All the analyses were repeated three times.Normal fasting rates of triglycerides range from 50 to 160 mg/dl; normal fasting rates of cholesterol range from 100 to 200 mg/dl .All the subjects' rates of cholesterol and triglycerides were compared to the maximum cut-off of the general population thus creating two ratios: cholesterol index (CI) and triglycerides index (TI).Through the operation CI - TI the LI was obtained. A positive index number means that the cholesterol production is prevalent, while a negative one indicates a prevalent triglycerides production.A value of + 0,15 or - 0,15 could be taken as 0, since it could be considered an irrelevant nutritional variation.The Statistical Package for Social Sciences was used. T-test and Z test were used. The association between categorical descriptive variables was tested through paired-samples t-test.Mann-Whitney test was used to compare the results regarding the Ewing score. Pearson correlation has been performed to find possible correlations between HRV and lipid metabolism. The p values were then obtained with t-test. Values of p \u22640.05, < 0.01 and < 0.001 were considered statistically significant.The anxious-depressive disorder group, compared to all the other groups , and to the control group, presented lower levels of cholesterol.With regards to the other groups, their cholesterol levels were higher compared to the control group, but there was not a statistically significant difference between the pathological groups compared among each other.The anxiety and anxious-depressive disorder groups, compared among each other, did not present a statistically significant difference in the triglycerides levels, but they both presented significantly higher triglycerides levels compared to all the other groups and to the control group ; .The depression group presented lower triglycerides levels, when compared to anxiety, anxious-depressive disorder and control groups ; higher triglycerides levels, when compared to the eating disorders group , which presented lower triglycerides levels than in all the pathological and control groups .The eating disorders group presented the highest LI among the pathological and control groups . The LI, among the anxiety and anxious-depressive disorder groups, did not present a statistically significant difference, but it was significantly lower than that of the control group and of the other pathological groups . In terms of average, the anxious-depressive disorder group presented the lowest LI of the whole sample of the study.The depression group presented, therefore, a higher LI than did the anxiety and anxious-depressive disorder groups , but did not present a statistically significant difference compared to the control group.The general population maximum cut-off was not exceeded either in the pathological groups or in the control one. Nevertheless, the anxious-depressive disorder group presented higher levels of Apolipoproteins A, compared to the other groups and to the control group. This same pattern was observed for Apolipoproteins B, only in the anxiety group.For more details about the results regarding lipid metabolism, see Table Among the pathological groups, the most represented autonomic dysfunctions were at a mild-mild/moderate level . When compared to the control group, only the depression and anxiety groups presented a statistically significant difference . The anxious-depressive group, in terms of mean value, is the group with the lowest HRV of the whole sample . Considering the whole sample, the correlation between ANS and lipid metabolism has been investigated, referring in particular to HRV, with highly statistically significant results . HRV and LI presented a positive correlation: the anxious-depressive disorder group, in fact, presented the lowest LI (showing to be the group with the most severe triglycerides hyperproduction) and the lowest HRV. Correspondingly, this group was the one with the highest average of Ewing score. HRV has been found to be inversely related to the triglycerides levels and positively related to the cholesterol levels; these results confirmed the HRV-LI correlation. Apolipoproteins A presented a negative correlation to HRV. The opposite has been found for Apolipoproteins B. For more details, see Table Lipid metabolism and autonomic functioning seem to be related to the discussed psychiatric disorders.In a context of enhancing knowledge, the lipid index could represent another biomarker to be taken into consideration.Our findings are far from being definitive, also considering the small sample size and some disparities in male and female distribution. Further studies are necessary.LI: Lipid index; GPs: general practitioners; DSM- IV-TR: Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision; HRV: Heart rate variability; ED: Eating Disorders; ANS: Autonomic nervous system; CI: Cholesterol index; TI: Triglycerides index.The authors declare that they have no competing interests.EP collected data, interpreted the results, and helped draft the manuscript.ML, AL, VM assisted with collecting data, interpreting the results and drafting the manuscript. CC designed the study, interpreted the results, and helped draft the manuscript. All authors read and approved the final manuscript."} +{"text": "Alaska Native Medical Center: A History 1953\u20131983. Anchorage: Alaska Native Medical Center, 1986. 2nd printing 2012Robert Fortuine. The late Dr. Robert Fortuine (1934\u20132009) had a distinguished career in Alaska as a Commissioned Officer of the US Public Health Service. His passion was medical history, and he published several books on the history of health and health care in Alaska.th anniversary of the hospital. Now, through the efforts of Susan Clift and Patricia Hackley, and financial support from the Alaska Native Tribal Health Consortium, the ANMC Auxiliary, and many individuals, a second printing has been published online. It can be purchased from Lulu.com:Long out-of-print and unavailable, this history of the ANMC was written to commemorate the 30http://www.lulu.com/shop/search.ep?type=Print+Products&keyWords=fortuine&sitesearch=lulu.com&q=&x=10&y=8Circumpolar Health Atlas. Toronto: University of Toronto Press [ISBN 978-1-4426-4456-4]T. Kue Young, Rajiv Rawat, Winfried Dallmann, Susan Chatwood, and Peter Bjerregaard, editors. This is an atlas about the health of the diverse populations who inhabit the circumpolar regions in the northern hemisphere. As an atlas, it uses maps, charts, tables, and images to describe and explain visually the major health patterns and related issues. The editors define and conceptualize \u201chealth\u201d very broadly, and it is also their conviction that health researchers, service providers, and policy makers working in the North and for the North need a broad and multidisciplinary understanding of northern conditions to put health into its proper context. There are 5 parts: (1) The Circumpolar World; (2) Circumpolar Peoples; (3) Health Status; (4) Health Determinants; and (5) Health Systems.The book can be purchased from the University of Toronto Press and major online booksellers:http://www.utppublishing.com/Circumpolar-Health-Atlas.htmlHealing Histories: Stories from Canada's Indian Hospitals. Edmonton: University of Alberta Press, 2013 [ISBN 978-0-88864-650-7]Laurie Meijer Drees. Healing Histories is a detailed collection of Aboriginalperspectives on the history of tuberculosis in Canada's indigenous communities and on the federal government's IndianHealth Services. Featuring oral accounts from patients, families,and workers who experienced Canada's Indian Hospital System, it presents a fresh perspective on health care history that includesthe diverse voices and insights of the many people affected bytuberculosis and its treatment in the mid-twentieth century. Healing Histories is essential reading for those interested in Canadian Aboriginalhistory, history of medicine and nursing, and oral history.The book can be ordered from:http://www.uap.ualberta.ca/UAP.asp?LID=41&bookID=1035"} +{"text": "AbstractAdephaga and the comparative anatomy of carabid coxal cavities. Ross and Joyce inspired several generations of students at UVM to take up advanced work in entomology and natural history.The lives and contributions of Ross and Joyce Bell are described with particular attention to studies of invertebrate natural history in the state of Vermont and carabid beetles of several groups, including the world rhysodine fauna. Their work, all done at the University of Vermont, was mainly taxonomic in nature and included aspects of the biology of the species considered. During their careers they described more than 75% of the c. 340 rhysodine species known to science. Ross Bell also wrote a number of seminal papers about the basal relationships of the PageBreakJune 2010, to honor them and recognize their contributions to local natural history in particular, to systematic entomology in general. In the printed Festschrift it is appropriate to provide a brief summary of their lives and their work in entomology. Ross Taylor Bell was born in Champaign, Illinois, on 23 April 1929. His father, Alfred Hannam Bell, was from Ontario, Canada, and studied Geology at the Universities of Toronto (B.Sc.) and Chicago (Ph.D.). He was an influential and productive petroleum geologist with the Illinois Geological Survey in Urbana. Alfred Bell\u2019s second wife, Dorothy Becker of Cincinnati, Ohio, was also a geologist employed at the Survey, having received her B.S. in Geology at the University of Cincinnati. Ross\u2019 parents met after the early death of Alfred\u2019s first wife from tuberculosis. They had three children, Ross and his two sisters Martha and Enid. By all accounts they were a family of enthusiastic naturalists. Both Martha and Enid married students who were contemporaries of Ross in the Department of Zoology at the University of Illinois and, furthermore, Martha became a professional botanist. Family vacations, which included long drives to various points in North America, were invariably a mix of botany, zoology and geology, including all the \u2018stuff\u2019 of classical natural history.Preceding production of this Festschrift, long-time friends and associates of Joyce and Ross Bell assembled at the University of Vermont (UVM) in Burlington during 10-13 Ross Bell\u2019s interest in insects began with the childhood gift of an insect collecting kit from his parents. This interest was exacerbated when someone gave him a jar full of silk worm larvae, which he reared on mulberry leaves, incidentally he claims developing his skills as a tree climber. At age 14, Ross got a job at the Natural History Survey at the University of Illinois. In this capacity he trapped flies and rose to the challenge of sorting sarcophagids, muscids and calliphorids and identifying them. The next summer he accepted a position with the taxonomic survey that he thought would take him toward his new found passion for insect identification, but instead he ended up washing glassware, installing moth balls in the collection and gathering box elder leaves for some reason that can no longer be recollected. Because this was during WW II, Ross posits that his eclectic roster of tasks resulted from a shortage of grad students for \u2018slave labor\u2019. Very likely these experiences had something to do with the respectful way that he subsequently treated his own students, always encouraging and supporting them to follow their own noses forward, knowing all along that a professor was interested in both them and their scientific findings.PageBreakRoss went to High School at the University Laboratory High School in Champaign, which he recalls as a magnificent place -- and so it must have been because it produced three Nobel Prize Laureates, two of them in science. Ross\u2019s mother always thought that he should\u2019ve done the same, but alas, as Ross pointed out in self-defense, no such prize was awarded in the category of \u2018carabidology\u2019. Ross spent the halcyon summers of his youth at his Aunt and Uncle\u2019s farm in West Alexandria, Ohio, helping with farm chores, and at every chance collecting and attempting to name insects from the fields and a stream which bordered the east boundary of the farm. This was his first bash with aquatic insects, something that remained a side interest throughout his career. In his last year of high school he rode his balloon-tired bicycle all the way to West Alexandria. It required two days of pedaling and an overnight at a hotel in Noblesville, Indiana. This 215 mile bicycle ride was an achievement that gave Ross much pleasure, both in the doing and in the subsequent telling.Carabidae (Simplicia) of Illinois. Between 1950-53 he completed a doctoral dissertation about the comparative morphology and phylogeny of Adephaga, under the sponsorship of the well-known entomologist, W. V. Balduf, whose early pioneering work in \u2018bionomics\u2019 helped establish the scientific basis for insect ecology in North America. Balduf introduced Ross to Blatchley\u2019s Coleoptera of Indiana in exchange for Ross\u2019s efforts in sorting and counting rose hips for one of Balduf\u2019s studies. Ross became much interested in the proper application of names through exposure to the plant taxonomist E. Neville Jones. With this motivation, Blatchley\u2019s useful keys allowed Ross to confidently name his catches and lured him into the world of beetles . His resulting thesis work on the Adephaga is recognized as a classic among carabidologists and it led to a host of publications on the topic . One of Ross\u2019s best friends during his time at Illinois was \u201cButterfly\u201d Bob Snetsinger, who became an economic entomologist at Penn State. Spurred on by interactions with Bob, Ross developed interests in life history and larval biology , and passed them on enthusiastically to his students . While at Illinois, Ross also became interested in ecology, was inspired by Professors Victor Shelford, S. Charles Kendeigh and Arthur Vestal, and served as President of the department\u2019s Ecology Club.Ross spent the immediate post-war years (1946-49) at the University of Illinois where he earned a B.S. in Zoology. In one additional year, he earned an M.S. with a thesis about the At graduation and with Ph.D. in hand, Ross was awarded a Fulbright Fellowship to go to India. After receiving his boat ticket he was packing to sail in two weeks when he received a letter from the Selective Service Board ordering him to appear instead for military duty. Thus he got an unexpected all-expense paid trip to Fort Leonard Wood in the winter for basic training. After his basic training he was sent to Fort Dietrich, MD, then known as the nation\u2019s \u2018Germ Warfare Center\u2019. Here he spent two years in the company of fleas. He regards his main accomplishment as discovery of a fast way of distinguishing males from females during the sorting of samples; males wear pinstripes, females do not. Upon discharge from the US Army\u2019s flea sorting brigade in 1955, Ross was hired by the Department of Zoology at the University of Vermont to fill a one-year position. The University realized that they had a \u2018catch\u2019 and offered him a permanent tenured position a year later. He spent his entire career there as a popular professor contributing excellent service to the University and teaching inspirational courses in Field Zoology, Invertebrate Zoology, Entomology and a summer course for graduate students in Mountain Ecology. During this period he inspired numerous undergraduate students, guided seven graduate students over the course, pursued taxonomic and natural history research on the New England and Vermont faunas and studied rhysodine beetles on a worldwide basis. For most of his career Ross was the Zoology Department\u2019s main strength in systematics and natural history as it connected to ecology and evolutionary biology.PageBreakcompanion at the University of Illinois, who had gone on to teach entomology at the University of Colorado. It was on this trip that Ross found his first undescribed rhysodine beetle species, and so began a thread of passionate discovery that established him as the world\u2019s expert on these interesting beetles . In the following year Ross collected a new entomological companion through marriage to Joyce Elaine Rockenbach of Whitestone, Queens, New York City. Joyce, who had completed her B.S. at Queen\u2019s College, had come to UVM to pursue a Master\u2019s degree after eight years of being a research assistant at the Columbia College of Physicians and Surgeons. Interestingly, Joyce\u2019s graduate advisor at UVM was Reuben Torch, who had been Ross\u2019 assistant lab instructor at the University of Illinois. After ten years of teaching nurses in the UVM School of Nursing, Joyce turned her fulltime attention to entomological pursuits, jointly with Ross. They became inseparable companions in the pursuit of entomology -- especially of carabid lore \u2013 and through these endeavors also became well-known pillars of the Vermont naturalists community. As Ross\u2019 eyesight waned, Joyce became his eyes and illustrator for the taxonomic work. Her skills as an excellent microscopist were much appreciated by Ross and his students. Although her grandfather had told her that \u2018a woman can either teach or be a nurse\u2019, Joyce Bell became an excellent entomologist in her own right.Shortly after joining UVM, Ross began a program of summer collecting followed by taxonomic work that would characterize his research career. He spent the summer of 1956 collecting insects in Mexico with Don Van Horn, a graduate student The newlyweds went on a honeymoon \u2018collecting\u2019 trip to Bar Harbor, during which they recall that they spent more time making car repairs than collecting beetles. The following summer they collected on Cape Breton, Nova Scotia, and also in the Great Smoky Mountains, where they met career-long \u2018beetle buddies\u2019 Tom Barr and Willy Rosenberg. He and Joyce returned to Mexico in the summer of 1959, driving from Burlington to Chiapas where they stayed with Franz Blom in San Cristobal. Franz was an archeologist-anthropologist, who worked with the indigenous people of the area, and his wife, Trudy, was a Swiss photographer studying the Lacondon Indians of the forests in the area. Franz had purchased a monastery and used it as his home and research base. He generously took in grad students, mainly in anthropology, and mainly from the University of Michigan, but also opened his doors to other people who wanted to study in the region. A stay in the monastery came with room and board and free evening fireside chats. The food, fondly recalled as \u2018tasty Mexican dishes, including memorable stuffed squash blossoms\u2019, came mainly from the Blom garden. During their return to Vermont, Ross and Joyce stopped in Arizona near Nogales in the Chiricahua Mountains where they met for the first time George and Kay Ball, their 2 sons and Ron Madge. So began another career-long association and beetle intensive interaction.PageBreakrmont. During three summers spent in Stowe, they collected extensively on and around Mt. Mansfield, the state\u2019s highest mountain. They also spent summers collecting in northeastern Vermont around Lake Willoughby, in southern Vermont based near Manchester and at other locations in the state. Through this work they built the UVM Entomological Collection into a significant resource for northern New England. In Victory Bog on the Moose River near Lake Willoughby, Andy Moldenke and the Bells first collected Bembidion bellorum, eventually described and named in their honor by David Maddison.During the 1960s the Bells began a program to learn the fauna of Vermont and to compile extensive records of natural history information. Their goal, as explained to us, was to provide a scientific basis for the study of arthropods in Vermont and to facilitate proper identification of insect specimens. Ross and Joyce developed a system of establishing a base camp of operation in an area and then collecting extensively in that general locale. For example, over a span of eight years they rented \u2018summer houses\u2019 around VePlatypatrobus lacustris. This was an exciting find because the beetle was known from only two puzzling specimens and its habits were virtually unknown. Once advised of this capture, Phil Darlington and his wife, Libby, drove immediately from Harvard up to Vermont and they all invested much energy in searching for the habitat of this rare catch. Unfortunately they were not successful; however, the story did not end there, but instead the mystery was solved through the web of carabidological connections in which Ross and Joyce were enmeshed, as elaborated below.The Bells frequently ran light traps during these summer evenings, and this activity contributed much to the excitement of the chase. One night, for example, the light trapping garnered three specimens of Platypatrobus lacustris lived in beaver houses. Despite having extensively searched the beaver meadow near the light trap location, the Bells and Darlingtons had not thought to disassemble the house itself and so the discovery eluded them. During the Stowe summers Ross and Joyce also met Ken Cooper, then at Dartmouth. Although a wasp specialist, Ken was also interested in carabids, and a number of techniques that Ken developed, including methodology for everting the internal sac of carabid males, made their way into the playbooks used by Ross and his students. During a summer spent in southern Vermont, they met Carl Parsons, an ex-UVM professor who had abandoned academe to open a bookstore in Manchester. Parsons was an avid collector who willed his excellent Vermont collection to the University, and thus the room that houses the entomological collections in historic Torrey Hall at UVM was named in his honor.Pursuit of their \u2018naturalist\u2019 activities led to numerous intersections between the Bells and others of entomological persuasion, especially those interested in ground beetles. These connections were of great benefit, interest and inspiration to Ross and his students. For example, in 1967 Carl Lindroth, the famous Swedish entomologist and carabid aficionado, visited the Bells and gave a talk at UVM about his work on the faunal development of Surtsey Island, Iceland, after its then recent volcanic uprising from the ocean floor. This drew three Quebec entomologists to Vermont and provided the first connection between the Bells and Andr\u00e9 Larochelle and Henri Goulet. Only later, however, did Henri Goulet discover that PageBreaksociety\u2019s summer field trips and provided much input for the VES Newsletter. This newsletter has become a heartbeat for entomology in Vermont and it is edited by Trish Hanson, one of Ross\u2019 graduate students and who is now an entomologist with the Vermont Forest, Parks and Recreation Department.Although this valuable aspect of the careers of academics is frequently underestimated, interested people with active research programs become the nodes of scholarly interaction that link and encourage development of research communities. Ross and Joyce Bell were for many years a significant link in North America\u2019s carabidological community, and a central node for entomology in the State of Vermont. They have been mainstays of the Vermont Entomological Society (VES) founded in 1992. Ross served as one of the first Presidents of the VES, and the Bells participated in the Carabidae of the area (Bell 1971) as well as a key to the darkling beetles of the area. In 1974 the Bells joined newly-hired UVM colleague and mammalogist Charles Woods on a trip to Haiti. While Charles looked for mammals, the Bells enriched their rhysodine collection and were drawn further into the mysterious tunnels of rhysodine biology. In 1982 Ross had a sabbatical which took Joyce and he to New Zealand and Papua New Guinea, where they worked out of the Wau Ecological Institute in the northeastern New Guinea Highlands. They were introduced to the Institute by Lindsay and Peg Gressitt of the Bernice P. Bishop Museum, who were killed tragically later that year in a plane crash in China. In addition to fending off burglaries by natives, they spent many days climbing Mt. Missim searching for rhysodines and other entomological treasures. One of those rhysodines was an undescribed species, named by Ross as Omoglymmius (s. str.) gressitti in honor of Gressitt. Their \u2018guard\u2019 at Wau was a reformed cannibal who sported a bone through his nose and carried a spear, and Joyce spent many a sleepless night fearing that the guard might have a relapse in habit.The 1970\u2019s and 1980\u2019s were very busy but stimulating years for Ross and Joyce, partly because they extended the geographical boundaries for their entomological work beyond Vermont. For example, in 1970 Ross was hired for the summer to work on the NSF \u2018Biome Project\u2019 on the short grass prairie of Pawnee, Colorado. Ross produced keys to In 1989-90 Ross took a second sabbatical during which he and Joyce were \u2018down under\u2019 in Australia, this time headquartered in Canberra with the CSIRO . During this trip they undertook two extensive collecting trips in search of rhysodines, one to Tasmania and the other to Queensland. The brilliant carabidologist, Barry Moore, was their host, friend and tour guide for the year. Ross and Joyce fondly recall his enormous contributions to their understanding of Australian natural history. Upon their return from the last trip to Australia, Jonathan Leonard proposed that he and Ross should write a book on Tiger beetles of the Northeastern United States. Thus, the \u201cNortheastern Tiger Beetles\u201d (Leonard and Bell 1999) was born as the definitive reference to these beetles, with color plates of all the species and including keys for third instar larval identification.PageBreak married and immigrated to Tasmania where Mike took a job teaching High School Biology in Huonville. Mike was recently awarded Australia\u2019s highest award for high school teaching and remains active as a buprestid enthusiast and collector, contributing among other things Tasmanian carabids of interest to the work of Bob Davidson at CMNH \u2013 a further example of continuing entomological threads starting with the Bells. Students fortunate to be part of the summer collecting blitzes learned the fascination of finding new things and learning to put names on them effectively. In the big picture they also learned about how such activities form the basis for zoological research that connects to what people can observe directly in their own backyards. There were ancillary lessons; e.g., that gin and tonic is not just some high-brow concoction, but also a great pre-dinner drink on a sultry July evening, or that Joyce Bell was an excellent and adventurous cook, in addition to having rare talents for making unusual entomological discoveries!Students at UVM were attracted to entomological work because of Ross\u2019 informative teaching style, filled with anecdotes about the doing of natural history, and his clear philosophy of helping others learn how to construct and pursue their own dreams. He was among a handful of professors at UVM during those times whose research work connected to field biology and natural history, and thus was a lightning rod for many students interested in these areas. The Bells generously involved many students in their summer research adventures. For example, the late Bob Mills, who was an accomplished Biology teacher at Putney School, was an able, enthusiastic assistant during the summers in Stowe. Mike Bouffard also worked with the Bells as an undergraduate student. He and Reni Shangraw, another student who worked in Biology with Ross,Chlaenius also flowed downstream from work that Ross had done . Bob, who began as an English major at UVM, was coaxed into entomology through exposure to Ross\u2019 course, and began to collect and study carabids as an undergraduate. Encouraged by the Bells, he continued to collect during his three years in the Peace Corps in Nepal (many new carabids subsequently described), and during this time was invited to return to UVM as Ross\u2019 graduate student in entomology, which led to a successful career as a coleopterist and curator at the Carnegie Museum of Natural History. John Spence\u2019s Master\u2019s work on Nebria grew out of a discussion that linked Ross\u2019 first publication (Bell 1955) to observations of the day on the rocky banks of Gleason Brook. This work and Denise Martin Leonard\u2019s M.S. project about stoneflies involved periodic visits to many stations along the length of the stream at different elevations, and through active participation in these research projects, Ross and Joyce came to know virtually every inch of \u2018the Brook\u2019. In fact, Gleason Brook and the adjacent North-facing slopes of Camel\u2019s Hump loom large in Ross\u2019 life as a special area for both teaching and research. Other students, including Brian Farrell, Jonathan Leonard, John Strazanac, and Peter Wimmer participated in the \u2018Bell natural history blitzes\u2019 and went on to successful careers in entomology.Many of the things that Ross pursued were eventually connected to \u2013 or even expanded in \u2013 work done by his students. For example, Bart Chiolino, a promising Vermont student, worked as Ross\u2019 assistant during the summer in Colorado, before completing an M.S. on wing-dimorphism in ground beetles. During the summer spent in the area of Lake Willoughby they were joined by Andy Moldenke, a summer research student on leave from Wesleyan College. Moldenke subsequently earned a doctorate in entomology at Stanford University and spent a productive career as a soil biologist at Oregon State University. Bob Davidson\u2019s extensive treatments of PageBreakBrian Farrell undertook an all-taxon biotic inventory of the Boston Harbor Islands, the carabid portion of which was done in collaboration with Davidson and Bell. Other graduates of the Field Naturalist program who pursued entomological interests include Jeff Collins, Susan Morgan and Mark Ward.Ross\u2019 Invertebrate Zoology course became especially popular with graduate students in the Field Naturalist Program at UVM in the 1990s, and this experience inspired Jessica Rykken, for example, to study the role of carabids as indicators of land types in the Green Mountains. She then followed the Bell network west to study carabids and other invertebrates in riparian areas with Andy Moldenke, and later with China and SE Asia. Most recently, Ross has recognized specimens that David Kavanaugh collected in Yunnan as belonging to three new species of three rhysodine genera (Bell and Bell 2011). Ross continues scientific activity as Research Associate with the Carnegie Museum in Pittsburgh. Ross has now turned his attention to the perplexing matters of rhysodine zoogeography, hoping to find some sensible patterns in the wealth of new data that he and Joyce have generated. They have more than quadrupled our knowledge of rhysodines by adding description of c. 260 new species to the list of about 80 known when they started.Ross retired from UVM in 2000 but spent four subsequent summers teaching a field course in entomology at UVM. He continues his work with rhysodine beetles and many collaborators in Europe still send him specimens mainly from expeditions to pallipes group of Nebria in eastern United States . Proceedings of the Entomological Society of Washington 57: 265-267.Bell RT (1955) Species of the Carabus auratus L. in North America. Proceedings of the Entomological Society of Washington59: 254.Bell RT (1957) Chlaenius tomentosus (Say) . Annals of the Entomological Society of America 51: 432-435.Bell RT (1958) Intraspecific variation in Scaphinotus Dej., intermediate between Scaphinotus s. str. and Irichroa Newman . Proceedings of the Entomological Society of Washington 61: 11-13.Bell RT (1959) A new species of Chlaenius Bonelli in North America. Miscellaneous Publications of the Entomological Society of America1: 97-166.Bell RT (1960) A revision of the genus Bell, RT, Bell JR (1962) The taxonomic position of the Rhysodidae (Coleoptera). The Coleopterists Bulletin 16: 99-106.Gehringia belong to the Isochaeta? (Coleoptera: Carabidae). The Coleopterists Bulletin 18: 59-61.Bell RT (1964) Does Platypatrobus lacustris Darlington in Vermont (Coleoptera: Carabidae). Proceedings of the Entomological Society of Washington 66: 100.Bell, RT, Bell JR (1964) Bell RT (1965) Coxal cavities and the phylogeny of the Adephaga. Proceedings of the XIIth International Congress of Entomology, pp. 80-81.Trachypachus and the origin of the Hydradephaga (Coleoptera). The Coleopterists Bulletin 20: 107-112.Bell RT (1966a) Chlaenius patruelis LeConte a valid species (Coleoptera: Carabidae). Proceedings of the Entomological Society of Washington 68: 321-322.PageBreakBell RT (1966b) Bell RT (1967) Coxal cavities and the classification of the Adephaga (Coleoptera). Annals of the Entomological Society of America60: 101-107.Bell RT (1970) The Rhysodini of North America, Central America, and the West Indies (Coleoptera: Carabidae or Rhysodidae). Miscellaneous Publications of the Entomological Society of America6: 289-324.Bell RT (1971a) Carabidae (ground beetles). Grassland Biome, United States International Biological Program, Technical Report No. 66: 58 pages.Bell RT (1971b) Handbook of the Malacostraca of Vermont and neighboring regions. Privately printed. 65 pages.Clinidium from Guatemala . Proceedings of the Entomological Society of Washington 75: 279-282.Bell RT (1973) A new species of Omoglymmius Ganglbauer, a separate genus (Coleoptera: Carabidae or Rhysodidae). The Coleopterists Bulletin 29: 351-352.Bell RT (1975) Clinidium (Coleoptera: Rhysodidae or Carabidae) from eastern U. S., with a revised key to U. S. Clinidium. The Coleopterists Bulletin 29: 65-68.Bell RT Bell JR (1975) Two new taxa of Nebria lacustris (Casey) and Nebria pallipes (Say) . The Coleopterists Bulletin 30: 81-84.Spence JR, Bell RT , Bell JR (1976) The larvae of Bell RT (1977) Ergebnisse der Bhutan-Expedition 1972 des Naturhistorischen Museums in Basel Coleoptera: Fam. Rhysodidae. Entomologica Basiliensia 2: 151-158.Platynus opaculus LeConte (Coleoptera: Carabidae) in Vermont. Cordulia 3: 157-158.Davidson RL, Bell RT (1977) Note on the distribution and habitat of Nebria suturalis LeConte in Vermont (Coleoptera: Carabidae). Cordulia4: 82.Bell RT (1978) The habitat of Omoglymmius, subgenus Nitiglymmius, new subgenus (Coleoptera: Carabidae or Rhysodidae). Quaestiones Entomologicae 14: 43-88.Bell RT, Bell JR (1978) Rhysodini of the world. Part I. A new classification of the tribe, and a synopsis of Evarthrus sodalis sodalis LeConte in Vermont (Coleoptera: Carabidae). Cordulia 4:Bell RT, Nielsen GR (1978) Bell RT, Bell JR (1979) Rhysodini of the world. Part II. Revisions of the smaller genera (Coleoptera: Carabidae or Rhysodidae). Quaestiones Entomologicae 15: 377-446.Bell RT (1979) Zoogeography of Rhysodini--do beetles travel on driftwood? In: Erwin, TL, Ball GE, Whitehead DR, Halpern AL (Eds.). Carabid Beetles: their Evolution, Natural History and Classification. Proceedings of the First International Symposium of Carabidology, Smithsonian Institution, Washington, D. C., August 21, 23 and 25, 1976. Dr. W Junk bv Publishers, The Hague-Boston-London, 331-342.Bell RT, Bell JR (1981) Insects of Micronesia, Coleoptera, Rhysodidae. Bernice P. Bishop Museum Insects of Micronesia 15(2): 51-67.Omoglymmius Ganglbauer (Coleoptera: Carabidae or Rhysodidae) and substitutions for preoccupied generic names. Quaestiones Entomologicae 18: 127-259.Bell RT, Bell JR (1982) Rhysodini of the world. Part III. Revision of PageBreakBell RT (1985a) A catalog of the Coleoptera of America north of Mexico. Family: Rhysodidae. U. S. Department of Agriculture, Agriculture Handbook 529-4, fascicle 4: x + 4 pages.Bell RT (1985b) Zoogeography and ecology of New Guinea Rhysodini (Coleoptera: Carabidae). In: Ball, GE (Ed.). Taxonomy, Phylogeny and Zoogeography of Beetles and Ants. Dr. W. Junk Publishers, Dordrecht, 221-236.) Pentagonica of the West Indies (Coleoptera: Carabidae). The Coleopterists Bulletin 39: 321-327.Bell RT . Quaestiones Entomologicae 21: 1-172.Bell RT, Bell JR (1985) Rhysodini of the world. Part IV. Revisions of Pentagonica Schmidt-Goebel (Coleoptera: Carabidae) from the southwestern United States. The Coleopterists Bulletin 41: 373-376.Bell RT (1987) A new species of Clinidium Kirby (Coleoptera: Carabidae or Rhysodidae) from Mexico, and descriptions of the females of two Neotropical members of the genus. Annals of Carnegie Museum 56:193-196.Bell RT, Bell JR (1987a) A new species of Bell RT, Bell JR (1987b) A new subtribe, genus and species of Rhysodini from South Africa (Coleoptera: Carabidae or Rhysodidae). Journal of the Entomological Society of Southern Africa 50: 287-290.Yamatosa, and a major range extension for Omoglymmius sakuraii Nakane (Coleoptera: Carabidae or Rhysodidae). Revue suisse de Zoologie 94\u00a0: 683-686.Bell RT, Bell JR (1987c) Rhysodine beetles in the Geneva collection: a new species of Harpalus rubripes (Duftschmid), a European ground beetle new to North America (Coleoptera: Carabidae). The Coleopterists Bulletin 41: 56.Bell RT, Davidson RL (1987) Bell RT, Bell JR (1988) Rhysodini of Sulawesi and nearby islands (Coleoptera: Carabidae or Rhysodidae). Journal of the New York Entomological Society 96: 7-15.chimbu group of Scopodes (Coleoptera: Carabidae). The Coleopterists Bulletin 43: 157-161.Bell RT, Bell JR (1989a) Revision of the Yamatosa and Omoglymmius, descriptions of undescribed sexes in other species, and some major range extensions (Coleoptera: Carabidae or Rhysodidae). Revue suisse de Zoologie96\u00a0: 637-642.Bell RT, Bell JR (1989b) Rhysodine beetles in the Geneva collection II: new species of In: Dindal, DL (Ed.). Soil Biology Guide. John Wiley and Sons, New York. xx + 1,349 pages.Bell RT (1990) Chapter 35. Insecta: Coleoptera, Carabidae, Adults and Larvae. In: Stehr, FW (Ed.). Immature Insects. Volume 2. Kendall/Hunt Publishing Co., Dubuque, Iowa, 1053-1092.Bell RT (1991) Rhysodidae (Adephaga). Pages 304-305 Bell RT, Bell JR (1991) The Rhysodini of Australia (Insecta: Coleoptera: Carabidae or Rhysodidae). Annals of Carnegie Museum 60: 179-210.Bembidion obtusum (Coleoptera: Carabidae) in eastern North America. Entomological News102: 174-178.Bell RT, Hoebeke ER, Liebherr JK (1991) Revised distribution of the immigrant carabid In: Noonan, GR, GE Ball and NE Stork (Eds.). The Biogeography of Ground Beetles of Mountains and Islands. Intercept Ltd., Andover, United Kingdom, 43-52.PageBreakBell RT (1992) The carabid fauna of the New England mountains (Coleoptera: Carabidae). Omoglymmius (subgenera Omoglymmius and Pyxiglymmius). Annals of Carnegie Museum 62: 165-185.Bell RT, Bell JR (1993) Rhysodine beetles (Insecta: Coleoptera: Carabidae or Rhysodidae): new species, new data and revised keys to Bell RT (1994) Beetles than cannot bite: functional morphology of the head of adult rhysodines (Coleoptera: Carabidae or Rhysodidae). The Canadian Entomologist126: 667-672. doi: 10.4039/Ent126667-3Bell RT, Bell JR (1995) The Rhysodini (Insecta: Coleoptera: Carabidae) of Cuba. Annals of Carnegie Museum 64: 185-195.In: Ball, GE, A Casale and A Vigna Taglianti (Eds.) Phylogeny and Classification of Caraboidea (Coleoptera: Adephaga). Proceedings of a Symposium XX International Congress of Entomology. Atti, Museo regionale di Scienze naturale, Torino 261-272.Bell RT (1998) Where do the Rhysodini belong? http://tolweb.org/Rhysodini/67/1999.12.14. In: The Tree of Life Web Project, http://tolweb.org/ Bell RT (1999) Rhysodini. Wrinkled Bark Beetles. Version 14 December 1999. Leonard JG, Bell RT (1999) Northeastern Tiger Beetles: a field guide to the tiger beetles of New England and eastern Canada. CRC Press, Boca Raton, Florida. 176 pages.Bell RT, Bell JR (2000) Rhysodine beetles (Insecta: Coleoptera: Carabidae or Rhysodidae): new species, new data II. Annals of Carnegie Museum 69:69-91.Yamatosa Bell and Bell and Omoglymmius (Pyxiglymmius) Bell and Bell. Stuttgarter Beitr\u00e4ge zur Naturkunde, Serie A (Biologie), Nr. 636:1-7.Bell RT, Bell JR (2002) Two new species of Rhysodini (Coleoptera: Carabidae) with revised keys to Bell RT (2003) Family Rhysodidae Laporte, 1840. In: L\u00f6bl, I, and A. Smetana (Eds.). Catalogue of Palaearctic Coleoptera. Volume I. Archostemata - Myxophaga - Adephaga. Apollo Books, Stenstrup, 78.Bell RT, Bell JR (2009) Rhysodine beetles (Insecta: Coleoptera: Carabidae): new species, new data III. Annals of Carnegie Museum 78: 45-77. doi: 10.2992/007.078.0104Bell, RT (2010) Les Rhysodini de Guyane . Suppl. Bulletin de liason d\u2019ACOREP-France \u201cLes Coleopteriste\u201d, Tome II: 46-49.Grouvellina Bell & Bell and the mishmicus group of Rhyzodiastes Fairmaire. Stuttgarter Beitr\u00e4ge zur Naturkunde A, Neue Serie 4: 129-135.Bell RT, JR Bell (2011) Four new species of Rhysodini (Coleoptera: Carabidae) with revised keys to"} +{"text": "Journal of Applied Crystallography includes some highlights of the 11th Biennial Conference on High-Resolution X-ray Diffraction and Imaging (XTOP), held in St Petersburg in 2012.This issue of Journal of Applied Crystallography includes some highlights of the 2012 Conference on High-Resolution X-ray Diffraction and Imaging (XTOP). The 11th Biennial Conference was held in St Petersburg, Russia, by the Ioffe Physical Technical Institute of the Russian Academy of Sciences, the Russian National Committee for Crystallography, the A. V. Shubnikov Institute of Crystallography and the National Research Centre \u2018Kurchatov Institute\u2019. This successful conference series started in Marseille (1992), with subsequent meetings in Berlin, Palermo, Durham, Ustron-Jaszowiec, Grenoble, Prague, Karlsruhe, Linz and most recently Warwick in 2010.This special issue of etc., in addition to the conventional methods of X-ray diffraction analysis and topography. The development of research techniques has been facilitated by the increasingly wide use of synchrotron radiation and computer processing. As a consequence, topics devoted to X-ray optics, coherent imaging, time-resolved diffraction etc. have appeared in the programmes of the XTOP symposia.The original goal of this conference series was to discuss the development of modern techniques in X-ray diffraction analysis and the application of these methods to study structural properties of crystalline objects. The conference subject matter has been steadily expanding to include X-ray standing waves, refraction tomography, X-ray reflectometry in situ measurements on colloidal crystals, to epitaxial layer characterization , nanoparticles and nanowires, and studies on defects and crack propagation. There are also articles on the theory of diffraction from distorted crystals, methods in micro-tomography and its use in biological studies, and new ideas in crystal optics to condition X-rays to improve data for the future.The range of the objects under study has expanded and now includes single crystals, surface crystalline layers, nanostructures and biomaterials. The technological development of new materials and structures, including nano-size systems and particles, has required a detailed and exhaustive knowledge of the structural state of these objects, which has stimulated further intensive development of X-ray research techniques. It is the solution to these challenges that was the essence of XTOP 2012. The highlights presented here include a good cross section of the activities in this field, from ordering in polymers, through Over 220 participants from more than 20 countries attended this conference, which lasted for four-and-a-half days and included six tutorial lectures, nine invited talks, about 60 oral presentations and three poster sessions with about 150 posters. The highlights included in this special issue give a representative view of the conference series and those four-and-a-half days in St Petersburg in September 2012."} +{"text": "L-DOPA is still regarded as the standard pharmacotherapy for the treatment of the motor symptoms of Parkinson's disease (PD). However, the efficacy of this drug is limited by the emergence of dystonic and choreic involuntary movements, generally referred to as dyskinesia. Current interventions to treat dyskinesia are mainly based on continuous delivery of L-DOPA, administration of glutamatergic drugs , replacement or combined administration of L-DOPA with less dyskinetic, albeit less effective, dopaminergic agonists, and deep-brain stimulation of discrete regions of the basal ganglia.Preclinical research, on the other hand, is searching for novel approaches to the treatment of L-DOPA-induced dyskinesia, targeting alternative neurotransmitter systems, such as the serotonin, adenosine, and opioid systems, or identifying abnormalities in intracellular signal transduction and synaptic plasticity associated to this condition. This Special Issue discusses recent breakthroughs in this direction and, at the same time, provides an update of the clinical features and management of L-DOPA-induced dyskinesia.The article by J. Guridi et al. describes the mechanisms underlying the various forms of dyskinesia produced by prolonged administration of L-DOPA. Basic pathophysiological mechanisms and current treatments are presented in detail and critically discussed.I. Aviles-Olmos et al. focus on the comparison between L-DOPA-induced dyskinesia and graft-induced dyskinesia, an analogous condition observed in response to transplantation of fetal dopaminergic cells in PD patients. This article also provides a timely discussion of the use of gene therapy in PD and of its effects on dyskinesia.2-type receptors, in the treatment of dyskinesia.The article by N. Tambasco et al. presents the clinical and epidemiological characteristics of dyskinesia and describes the management of this condition, based on the use of various types of dopaminergic agonists. On the same line, J. C. P. Piedad and A. E. Cavanna discuss more in detail the use of pramipexole, an agonist at dopamine DA number of signaling components potentially implicated in dyskinesia have been discovered during the last decade. In this regard, V. Ghiglieri et al. provide a comprehensive overview of the mechanisms and the possible pathological consequences of abnormalities affecting various forms of corticostriatal plasticity associated to L-DOPA-induced dyskinesia.Recently, studies have pointed to the serotonergic system as a key player in the aberrant effects produced by L-DOPA and linked to the emergence of dyskinesia. The article by S. Navailles and P. De Deurwaerd\u00e8re describes the evidence at the basis of this hypothesis and discusses the use of therapeutic approaches designed to prevent this phenomenon. On the same subject, E. Shin et al. discuss the experimental and clinical evidence implicating the serotonin system in L-DOPA- and graft-induced dyskinesia.2A receptors have attracted considerable interest as antiparkinsonian and antidyskinetic agents. The article by M. Morelli et al. provides a critical appraisal of the potential therapeutic properties of these compounds, as determined in experimental models of PD and L-DOPA-induced dyskinesia.Drugs acting as antagonists at adenosine AFinally, G. Frazzitta et al. present an interesting study showing the beneficial effects produced on dyskinesia by intensive physiotherapeutic rehabilitation. These results are discussed with regard to similar studies performed in animal models of PD."} +{"text": "KORA-Age is a multidisciplinary research consortium funded by the German Federal Ministry of Education and Research (FKZ 01ET1003). The objective of KORA-Age is to examine the prevalence and determinants of mortality, multi-morbidity, functioning and successful ageing in persons aged 65 years or older in a large cohort of randomly selected inhabitants of the region of Augsburg, Germany.http://www.helmholtz-muenchen.de/en/kora-en/index.html). The cohort includes all participants of the four cross-sectional MONICA/KORA Surveys S1-S4 conducted between 1984 and 2001 who were born in or before 1943, in total 9,197 persons. Within KORA-Age, a mortality follow-up was conducted in 2008 and 2011. In 2008/2009, a mailed questionnaire was answered by 4,565 individuals and a telephone interview with 4,127 individuals conducted. A physical examination was performed in a gender and age-stratified random sample of the cohort in 2009 in 1,079 persons. They were followed up three years later and a re-examination took place in 822 persons. Information was obtained from interviews, a self-assessment form and health examinations including morbidity, a comprehensive assessment on mental health, health services utilisation, medication and a complete characterisation of ageing-related biological parameters like cardiovascular and pulmonary function, tests of balance and stability, bone density, body composition and measures of multi-morbidity, frailty, disability and participation. In 2013 a group of older citizens will take part in qualitative focus groups on the lived experience of neighbourhood and environment to evaluate participation in-depth in the region of Augsburg. The sampling area was geocoded.The KORA-Age project is carried out within the framework of KORA . We propose this approach for integrating functioning into a comprehensive view of the prevalence and determinants of multi-morbidity, disability and successful ageing in interaction with personal and environmental factors."} +{"text": "Combat-intense, lengthy, and multiple deployments in Iraq and Afghanistan have characterized the new millennium. The US military's all-volunteer force has never been better trained and technologically equipped to engage enemy combatants in multiple theaters of operations. Nonetheless, concerns over potential lasting effects of deployment on long-term health continue to mount and are yet to be elucidated. This report outlines how findings from the first 7 years of the Millennium Cohort Study have helped to address health concerns related to military service including deployments.The Millennium Cohort Study was designed in the late 1990s to address veteran and public concerns for the first time using prospectively collected health and behavioral data.Over 150 000 active-duty, reserve, and National Guard personnel from all service branches have enrolled, and more than 70% of the first 2 enrollment panels submitted at least 1 follow-up survey. Approximately half of the Cohort has deployed in support of operations in Iraq and Afghanistan.The Millennium Cohort Study is providing prospective data that will guide public health policymakers for years to come by exploring associations between military exposures and important health outcomes. Strategic studies aim to identify, reduce, and prevent adverse health outcomes that may be associated with military service, including those related to deployment. Soon after the end of the 1991 Gulf War, veterans began reporting symptoms and illnesses they perceived to be possibly related to exposures during deployment. However, research on illnesses related to the 1991 Gulf War was hampered by the non-availability of objective measurements on exposures at the individual level, a lack of baseline health data, and an inability to adequately control for potential confounding factors. Baseline health data prior to potential military exposures are critical to appropriately evaluate associations between deployment-related exposures and subsequent health outcomes. Previous research involving cross-sectional and case-control study designs suffered from recall bias ,3. The MHealth status is assessed at baseline using self-reported questionnaire data linked to supplemental data from various DoD administrative and health databases. Follow-up data are collected every 3 years through postal or Web-based surveys with periodic queries of the same electronic databases. The launch of the Millennium Cohort Study in July 2001 occurred just prior to terrorist attacks on the United States on September 11, 2001, and the start of military mobilization and engagement in multiple combat theaters. Consequently, a significant proportion of study participants have experienced military deployment, and the stage was set to better understand any health consequences of these deployments.The Millennium Cohort Study is a population-based study consisting of participants drawn from a randomly selected sample of all US military service members on rosters as of October 2000 (Panel 1), October 2003 (Panel 2), and October 2006 (Panel 3) ,5. PanelDemographic and military personnel data from the Defense Manpower Data Center (DMDC) were obtained for each cohort member. These data include sex, age, marital status, race/ethnicity, education, service branch , service component , occupation, and military pay grade .A number of early studies were planned and executed to establish the representativeness of the Cohort, understand response bias, assess reliability in reporting, ascertain mortality, compare Web- and paper-based responses, and validate self-reported health outcomes with automated health information. These foundation analyses have documented a cohort representative of the general US military, data quality and reliability which are excellent, and little to no evidence of response bias ,6-15.In order to examine possible response bias, prior hospitalization and ambulatory care experience of responders was compared with that of nonresponders . FindingThe degree of nonrandom agreement between self-reported Millennium Cohort survey data and electronic records has been examined using Kappa statistics, where a kappa between 0.8-1.0 indicates \"greater than substantial agreement,\" between 0.6-0.8 \"substantial agreement,\" between 0.4-0.6 \"moderate agreement,\" between 0.2-0.4 \"fair agreement,\" and between 0.0-0.2 \"slight or poor agreement\" . Strong A major objective when designing the Millennium Cohort Study was to describe deployment experiences and exposures among US service members and relate them temporally to subsequent health outcomes. Special emphasis was placed on obtaining information on vaccines, environmental exposures, and combat-related experiences.Electronic deployment data are obtained from DMDC and include in- and out-of-theater dates for current operations. The study questionnaire also uses 24 country and sea codes to assess self-reported date and location of deployment since 2001.Table Among preventive measures used to counter threats of biological warfare agents, most deployed US service members receive anthrax and smallpox vaccinations. The Anthrax Vaccine Immunization Program began in the late 1990s, while the smallpox vaccination program began in 2001 ,13, as wQuestions from the National Health Survey of Gulf War Era Veterans were incBecause deployment may lead to increased risk of exposure to environmental hazards, such as exposure to pesticides ,30, chemThe longstanding and continued combat operations in Iraq and Afghanistan have fueled ongoing concerns among veterans and the general public over unknown exposures and potential long-term health consequences of serving in the military and, in particular, of deployment. Mental health disorders and more subtle physical sequelae may affect both short-term and long-term functional capacity and quality of life in troops returning from deployment. Studies have reported significantly higher rates of mental health disorders, such as PTSD, major depression, and alcohol misuse, after deployment in support of combat operations in Iraq and Afghanistan, as well as the 1991 Gulf War and the Vietnam War ,34-40. FPrevious cross-sectional and serial cross-sectional studies have reported alcohol misuse among personnel returning from combat operations in Iraq and Afghanistan ,51. AlcoInvestigation of deployment and new-onset disordered eating and changes in weight between baseline and follow-up surveys showed no differences among men and women who deployed, with or without combat exposures, and those who did not deploy . When deIn addition to changes in health risk behaviors possibly associated with deployment-related stressors, psychiatric disorders, such as depression and PTSD, have also received significant attention. To investigate depression among Millennium Cohort participants, the study questionnaire includes a standard self-administered clinical instrument to evaluate mental disorders, the Primary Care Evaluation of Mental Disorders Patient Health Questionnaire , and selPTSD symptoms have been reported among as many as 30% of veterans following service in Vietnam and in as many as 10% of personnel returning from the 1991 Gulf War ,53-58. MOngoing prospective investigations are critical to increasing our understanding of PTSD and include exploring the time course of new-onset and persistent PTSD. Furthermore, identifying and testing potential interventions to speed recovery and promote resilience in military members facing new challenges is vitally important. Complementary and alternative therapies, physical exercise, and identification and treatment to minimize effects of mild to severe traumatic brain injury may mitigate symptoms of PTSD. The 2007-2008 survey cycle added a third longitudinal data point for further investigation of new onset, persistence, and resolution of PTSD symptoms and offer a better understanding of resilience factors among cohort members.Table Increasing evidence suggests that a higher proportion of service members deployed to Afghanistan and Iraq are suffering from head and brain injuries compared with prior conflicts ,62. ThesWhile injuries remain one of the most significant health problems of the armed services, it is the sequelae of injuries, especially musculoskeletal conditions resulting in permanent disability, that exact the greatest and most lasting toll on our service members . MusculoMilitary deployment has previously been identified as a risk factor for deaths due to external causes in general, and specifically due to motor vehicle crashes, following both the Vietnam War and the 1991 Gulf War -67. The As the Millennium Cohort Study moves toward the end of the first decade of data collection, longitudinal data points will be used to investigate chronic medical and psychiatric conditions and any short- or long-term impact of military deployment on the development or natural history of these conditions. Presence of chronic medical conditions will be identified using several different methods, including self-reported questionnaire responses and electronic DoD and Department of Veterans Affairs health care databases. Even after separation from military service, the Millennium Cohort survey instrument will be the main method of assessing new onset of chronic illnesses, since fewer than 20% of veterans use Veterans Affairs health care facilities. The younger age of the cohort limits our ability to identify exposure-health outcome associations for common causes of mortality, such as coronary heart disease or cancer, during early periods of follow-up. However, chronic conditions with earlier ages of onset should be detectable in relation to exposures of interest. These include recently investigated conditions such as asthma, hypertension, and diabetes, as well as planned future investigations of multiple sclerosis, inflammatory bowel disease, and infectious diseases such as hepatitis C. Similar to incidence estimates for diabetes from the CDC , newly sThe current study is a population-based, prospective cohort design that allows for baseline and multiple follow-up assessments on the same individuals. The cohort design, large sample size, and ability to prospectively follow-up individuals for over 20 years make the Millennium Cohort arguably one of the most ambitious and challenging studies undertaken in the era of modern epidemiology ,73. HoweThe concept for the Millennium Cohort Study arose from lingering postdeployment concerns following the 1991 Gulf War . The popPerhaps most importantly, the Millennium Cohort Study will continue to play an important role in defining long-term health consequences of military occupational exposures, specifically during times of combat deployment. Such exposures include psychological and environmental stressors, medical interventions, and other occupational factors that are unique to military populations . The heaThe authors declare that they have no competing interests.TS originated the study and supervised all aspects of its implementation. IJ, CL, and BS assisted with analysis of data. IJ, TH, CL, EB, BS, GG, TW, PA, GG, JR, and MR assisted with design and interpretation of data, drafting and revision of the manuscript. All authors read and approved the final manuscript.At the time of this study, Tyler Smith, Isabel Jacobson, Cynthia LeardMann, and Besa Smith were at the Department of Defense Center for Deployment Health Research, Naval Health Research Center, San Diego, CA. Tomoko Hooper was with the Department of Preventive Medicine and Biometrics, Uniformed Services University of the Health Sciences, Bethesda, MD. Edward Boyko was with the Seattle Epidemiologic Research and Information Center, Department of Veterans Affairs Puget Sound Healthcare System, Seattle, WA. Gary Gackstetter was at Analytic Services, Inc. (ANSER), Arlington, VA. Timothy Wells and James Riddle were with the Air Force Research Laboratory, Wright-Patterson Air Force Base, OH. Paul Amoroso was with Madigan Army Medical Center, Tacoma, WA. Gregory Gray was with the Department of Environmental and Global Health, College of Public Health and Health Professions, University of Florida, Gainesville, FL. Margaret Ryan was with the Naval Hospital Camp Pendleton, Camp Pendleton, CA.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/11/69/prepub"} +{"text": "There were errors in the funding section. The correct funding information is as follows:This work was supported by a grant from the Lundbeck Foundation (www.lundbeckfonden.dk) (grant number R93-A8391), by an ERC Starting Grant (281869\u2014elegansNeurocircuits), and by an ERC Starting Grant (260807 - HYPOXICMICRORNAS). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "To the Editor: A recent extensive study on global distribution of rubella virus genotypes by Zheng et al. vaccine was introduced in the national vaccination program for 15-month-old infants. Because of the rubella outbreak of 1993, the vaccination policy changed. In 1997, a second vaccination was recommended for 11- to 12-year-old children. However, another major rubella epidemic occurred in 1999, beginning in late December 1998 and lasting until May 1999, with a peak in the number of cases in January. During this period, 1,438 rubella cases were reported throughout Greece; 765 were in the northern part of the country. In previous rubella epidemics in Greece, children were most affected. However, in the 1999 epidemic, a higher incidence rate was observed mostly among 15- to 19-year-old persons (During the 1999 epidemic, oral samples were collected from patients within a week of the onset of symptoms. Most of the samples were sent to Colindale, London, for testing; a few of them were stored at \u201370\u00b0C in our laboratory at the School of Medicine, Aristotle University of Thessaloniki, Thessaloniki, Greece. In 1999, four universities in the United Kingdom reported cases of rubella infection. Greek students were attending all of these universities; the students had spent the Christmas holidays in Greece and then returned to the United Kingdom. The U.K. rubella strains were identical to those of the Greek epidemic strain (Zheng et al. (Recent data indicate that internationally circulating rubella viruses exist, even when vaccination programs are conducted. Comprehensive specimen collection and genotypic analysis are necessary to identify and track the emergence and spread of such genotypes."} +{"text": "Breast cancer is the most common cancer in women, representing 16% of all female cancers. According to the American Cancer Society, long-term cancer survival is defined as more than five years of survivorship since diagnosis, with approximately 2.5 million breast cancer survivors (BCS) in 2006. The long-term effects from breast cancer and its treatment have been shown to have positive and negative effects on both recovery and survivors' quality of life (QoL). The purpose of the study was to identify QoL instruments that have been validated in long-term BCS and to review the studies that have used the QoL instruments in this population.A systematic literature search was conducted from January 1990 to October 2010 using electronic databases. Instruments validated and used in BCS were included in the review. In addition, QoL studies in long-term BCS using the validated instruments were reviewed. The search was limited to studies in English language. Studies of BCS of less than five years after initial diagnosis, any clinical or review studies were excluded.The review identified a total of 12 instruments validated in long-term BCS. According to the QoL framework proposed by Ferrell and colleagues, three instruments evaluated all four domains of QoL. A review of the psychometric evaluation showed that Quality of Life in Adult Cancer Survivors Scale has acceptable reliability, validity, and responsiveness in long-term BCS compared to other disease-specific instruments. The review also yielded 19 studies that used these QoL instruments. The study results indicated that age-group, ethnicity, and type of treatment influenced different aspects of QoL.There is a significant impact of breast cancer on QoL in long-term BCS. The review can help researchers and clinicians select the most appropriate instruments to assess the changes in QoL in BCS. Breast cancer is the most common cancer in women, representing 16% of all female cancers . Most sie.g., fatigue, sleep disturbances, and pain) and psychological distress after diagnoses that adversely affect their overall QoL and survivorship [The long-term effects from breast cancer and its treatment have been shown to have positive and negative effects on both recovery and survivors' quality of life (QoL). Also, QoL outcomes vary across the breast cancer continuum including diagnosis at different stages of breast cancer, disease-free survivorship beyond the first course of primary treatment, long-term disease-free survivorship, and first recurrence of breast cancer . Accordiivorship .The implementation and use of improved diagnosis and treatment programs have resulted in decreased breast cancer mortality . Howeveret al [The problems resulting from breast cancer and its treatment are varied and complex. Ferrell et al proposedet al to identify QoL instruments validated and used in long-term BCS; (2) to provide a description of the instruments and their psychometric properties in long-term BCS; and (3) to provide a systematic review of studies that used the QoL instruments in long-term BCS.quality of life, health-related quality of life, measures, scales, questionnaires, breast cancer, breast carcinoma, breast cancer survivors, long-term breast cancer survivors, post-treatment, post-chemotherapy, post-radiation therapy, and post-surgery. The search was conducted to identify studies reporting the use of QoL instruments in the evaluation of breast cancer and its treatment in long-term BCS. The QoL instruments used in these studies were also identified. These QoL instruments were then reviewed for their validation in long-term BCS. The literature search process is illustrated in Figure Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines , a systeInstruments were included in the review if they were used in at least one long-term BCS study and their description and psychometric properties were reported in BCS population with a varying number of survivor years. All included instruments were identified as patient-reported outcome questionnaires measuring one or more aspects of QoL . The instruments that measured patient satisfaction or patient preference were excluded. Also, the instruments without any description of their development or validation were excluded from the review.In addition to the inclusion of instruments, the review also included studies in long-term BCS that used these validated instruments. The search was limited to studies in English language and the use of the English version of the QoL instrument. The ACS's definition of long-term survival (> 5 years since diagnosis) was used ; studiesThere are generally three types of QoL instruments: generic, disease-specific, and condition-specific. The generic instruments are designed to measure the complete spectrum of disease in various populations and are useful in comparing QoL changes across different diseases . The disThe information on psychometric properties of the instrument was also extracted. The QoL instruments are generally tested for two types of reliability: internal consistency reliability and test-retest reliability. The internal consistency reliability is measured as Cronbach's alpha (\u03b1) whereas the test-retest reliability is estimated as Pearson product-moment correlation coefficient (r). The validity refers to the degree to which an instrument measures the concept it is intended to measure. The responsiveness is the ability to detect change in health status over time .For the studies reporting the use of QoL instruments, the following information was collected: sample size, socio-demographic variables , medical variables (type of treatment and tumor stage at diagnosis), inclusion/exclusion criteria used in the selection of population for their studies, QoL instruments used, administration of QoL instrument, and survivor's QoL.A total of 12 QoL instruments (10 disease-specific and two condition-specific) were identified and included in the review -27. The et al., three of the identified disease-specific instruments evaluated all four domains of QoL and included items consistent with survivor's concerns [The instruments varied widely in number of items and domains, mode of administration, scaling and scoring, and psychometric properties. The different measures used in BCS and their complete description such as items, domains, scaling, scoring, and means of administration is provided in Table concerns ,24,25. Oconcerns -19,21,22concerns . There aconcerns ,27. In tThe psychometric properties of the instruments used in BCS are presented in Table Based on the literature search methodology, 19 studies were identified that reported the use of QoL instruments in long-term BCS -46. TablMost of the studies utilized two or more QoL instruments to assess different aspects of QoL in long-term BCS. Cancer-specific QoL instruments designed to measure physical and psychosocial domains were used by six studies ,34,36,46The results from the 19 studies are summarized into six categories, although some studies met the criteria for more than one category. These include overall QoL and effect of different demographic and medical variables on QoL.et al. [et al. [et al. [et al. [et al. [Five studies assessed the effect of breast cancer and its treatment on overall QoL among long-term BCS ,34,39,40et al. indicate [et al. assessedet al. [et al. [et al. [Three studies examined the relationship between life stage variables (survivor's age at diagnosis and number of years of post-diagnosis survivorship) and QoL ,33,36. C [et al. focused et al. [et al. [Two studies examined the effect of different age groups on QoL ,42. Sammet al. assessedet al. [et al. [et al. [et al. [Four studies evaluated the relationship between long-term effects of breast cancer on QoL and ethnicity ,40,43,44 [et al. also comet al. [et al. [et al. [et al. [Four studies evaluated the effect of breast cancer treatment on different aspects of QoL or overall QoL ,37,38,45et al. examinedet al. [et al. [Two studies evaluated the effect of non-pharmacological intervention on QoL among long-term BCS ,46. Dirket al. reportedThe transition of a patient with breast cancer from treatment phase to survivorship phase is an important aspect of cancer continuum. The number of BCS and the number of survivorship years will increase with further advancements in breast cancer treatments. However, these therapies have persistent side-effects and toxicity which have been shown to negatively impact a survivor's QoL . Thus, tTo date, QoL assessments in cancer survivors and in BCS in particular, have employed several valid instruments that assess different QoL dimensions. As indicated by Ferrell and colleagues, consideration of psychological, social, physical, and spiritual aspects of QoL is essential in understanding the long-term impact of breast cancer diagnosis and treatments. Furthermore, few instruments are available that specifically evaluate all the different aspects of QoL in BCS. Three cancer-specific instruments, QLACS, QLI-CV, and QOL-CS evaluate all four domains of QoL and include questions specifically relevant to BCS. The EORTC and FACT instruments are important for measuring cancer-specific concerns, but they lack some survivor-specific concerns such as fear of recurrence, compromise with physical problems, and psychosocial issues. These cancer-specific instruments have shown similar results related to survivor's QoL when used in different studies focusing on BCS. For instance, presence of physical problems such as fatigue, pain, and insomnia as well psychosocial problems such as anxiety, depression, and concerns about appearance were consistently measured by these instruments.Another important criterion for the selection of QoL instruments is to ensure that the instruments have good psychometric properties. Almost all the instruments in BCS have reported good psychometric properties such as reliability and validity. However, most of the instruments have not been tested for responsiveness, an important property for assessing the change in QoL over time. The concerns related to different aspects of QoL change as a survivor transitions to another year of survivorship. The change may be positive or negative based on the survivor's perceptions, expectations, and overall health. It is, thus, important for the instruments to capture the changes in patient outcomes that vary with time. Additionally, none of the generic measures have been validated in a breast cancer population, creating a need for validation of these measures for evaluating changes in self-reported general health status of patients with cancer as they transition from treatment phase to survivorship. The generic instruments provide insight into the complete spectrum of disease and are useful not only in comparing QoL changes across different populations but also across different diseases . There are certain domains of QoL that are more prominent in BCS. For instance, fatigue, pain, insomnia, depression, anxiety, and fear of recurrence are persistent concerns that affect survivors and these conditions could affect their activities of daily living, coping skills, and social and role functions. The review identified only two condition-specific instruments that have been validated in BCS. However, these instruments do not fully assess the concerns of the survivors resulting in a need for validating the existing condition-specific instruments. Also, there are some disease-specific instruments such as Breast Cancer Prevention Trial (BCPT) Symptoms Scale , Impact Another goal of this review was to evaluate the studies that assessed QoL in long-term BCS using validated instruments. These studies assessed inter-connectedness and the important aspects of Ferrell's QoL domains. Most of the studies showed that these domains are inter-related. The physical concerns such as sexuality and menopausal symptoms impact both psychological and social aspects of QoL. The social concerns including changes in self-concept as well as physical concerns negatively impact psychological well-being. The spiritual well-being helps in improving physical, psychological, and social well-being by providing strength to cope with negative effects of breast cancer and its treatment.The studies also evaluated specific concerns of BCS related to each domain. First, the physical domain of QoL was most commonly evaluated by the studies, with most frequent issues being fatigue, sleep problems, and pain. Additionally, lymphedema (swelling of arms) was reported only in women treated with breast cancer surgery. These physical problems are typical for breast cancer treatment and have long-lasting effects; becoming even more problematic with increase in age. Second, the psychological well-being was also most frequently evaluated, with major symptoms being emotional distress and depression resulting from concerns such as fear of recurrence, uncertainty, family distress, financial burden, and worries about appearance. Higher level of psychological distress was reported in Hispanic women. Third, the social well-being was affected by absence of sufficient social support. This was a major concern in women 50 years and older as well as in Hispanic women. Fourth, spiritual well-being is also an important aspect of QoL, but is least often reported by the studies. Spiritual well-being was evaluated by two studies, one reported post-treatment positive change in life, while other study reported good overall QoL in African-American BCS resulting from higher levels of spirituality.In addition, these studies assessed specific concerns of BCS related to different demographic and medical variables. The studies reported an overall improvement in QoL in BCS, but the findings varied considerably based on the demographic and medical variables. For instance, the BCS diagnosed at a younger age experience problems with social well-being compared to the BCS diagnosed at an older age experiencing problems with physical well-being. The Hispanic women experience a higher degree of uncertainty compared to women of other ethnic groups. Moreover, the survivor's concerns vary with type of breast cancer treatment. Presence of lymphedema is common for BCS treated with mastectomy and concerns such as fatigue and pain are common for BCS treated with chemotherapy. Thus, the knowledge of variability in survivor's concerns is important for clinicians and decision-makers to help guide proper follow-up care after completion of breast cancer treatment.Most of the studies have focused on QoL during breast cancer diagnosis and treatment, but there is a gap in the literature for studies focusing on long-term BCS. There is a need for studies comparing and evaluating the effects of different breast cancer therapies in long-term BCS. More studies are required comparing QoL in BCS diagnosed at different stages of breast cancer as the extent of treatment and its effects might vary at different stages of cancer. There is a need for studies assessing impact of co-morbid conditions in older (> 60 years) BCS. Moreover, the studies analyzing the problems resulting from cancer recurrence are important. The studies reported in this review are cross-sectional studies, thus do not provide change in QoL over time in BCS. These studies provide varied results in different groups of BCS, which makes it difficult to generalize these results to the general population of BCS. Most of the studies included in this review have used more than one instrument, in which one or two instruments have been validated in the breast cancer population; other instruments used have only been validated in general population. The spiritual outcomes are the least reported, and consideration of the spiritual domain in future studies is important.This review highlights the significant and persistent effects of breast cancer and its treatment on long-term BCS and the inter-connectedness of the psychological, social, physical, and spiritual aspects of QoL in BCS. Thus, clinicians and decision-makers need to understand the complexity of problems in long-term BCS to help guide proper follow-up care after the completion of breast cancer treatment. This review also provides useful insight into the unique concerns and needs of BCS, which can help researchers and clinicians select the most appropriate instruments to assess the changes in QoL. The use of validated instruments will not only provide valid data but also help improve the quality of care in long-term BCS.ACS: American Cancer Society; BCS: Breast Cancer Survivors; BDI: Beck Depression Inventory; BIRS: Body Image and Relationships Scale; BPI: Brief Pain Inventory; BSI: Brief Symptom Inventory; CALGB: Cancer and Leukemia Group B; CARES-SF: Cancer Rehabilitation Evaluation System Cancer-Short Form; CBT: Cognitive Behavioral Therapy; CES-D: Center for Epidemiologic Studies Depression Scale; DAS: Dyadic Adjustment Scale; EORTC QLQ-BR23: European Organization for Research and Treatment of Cancer-Breast Module; EORTC QLQ-C30: European Organization for Research and Treatment of Cancer; FACIT-SP: Functional Assessment of Chronic Illness Therapy-Spiritual Well Being Scale; FACT-B: Functional Assessment of Cancer Therapy-Breast; FACT-G: Functional Assessment of Cancer Therapy-General; FIQ: Fibromyalgia Impact Questionnaire; FSI: Fatigue Symptom Inventory; HADS: Hospital Anxiety and Depression Scale; IOC: Impact of Cancer Scale; IES: Impact of Events Scale; LES: Life Experience Survey; LSI: Life Satisfaction Index; ISEL: Interpersonal Support Evaluation List; LOT: Life Orientation Test; LOT-R: Life Orientation Test-Revised; LTQL: Long-term Quality of Life; MFSI: Multidimensional Fatigue Inventory; MPQ-SF: Short Form McGill Pain Questionnaire; MUIS-C: Mishel Uncertainty in Illness Scale Community; OARS: Older American Services and Resources Questionnaire; PAL: Physical Activity and Lymphedema Trial; PANAS: Positive and Negative Affect Scale; PCL-C: Posttraumatic Stress Disorder Checklist-Civilian; POMS: Profile of Mood States; QLACS: Quality of Life in Adult Cancer Survivors Scale; QLI-CV: Quality of Life Index-Cancer Version; QOL: Quality of Life; QOL-CS: Quality of Life-Cancer Survivors; QOLS: Quality of Life Scale; SF-36: Medical Outcomes Study 36-item Short Form Health Survey; SDS: Symptom Distress Scale; SPS: Spiritual Perspective Scale; SSQ: Social Support Questionnaire; STAI: State-Trait Anxiety Inventory.The authors declare that they have no competing interests.IC contributed to the conception, design, collection and analysis of studies, and organized the complete draft. KK conducted a critical review of the manuscript for important intellectual content and provided finishing touch to the manuscript. Both authors read and approved the final manuscript."} +{"text": "Biosensors for detecting dioxins and related compounds, although still under development, show potential to overcome these limitations. Immunosensors and biomimetic-based biosensors potentially offer increased selectivity and sensitivity for dioxin and DL-PCB detection, while whole cell-based biosensors present interpretable biological results. The main shortcoming of current biosensors, however, is their detection level: this may be insufficient as limits for dioxins and DL-PCBs for food and feedstuffs are in pg per gram level. In addition, these contaminants are normally present in fat, a difficult matrix for biosensor detection. Therefore, simple and efficient extraction and clean-up procedures are required which may enable biosensors to detect dioxins and DL-PCBs contamination along the food chain.Dioxins and dioxin-like polychlorinated biphenyls (DL-PCBs) are hazardous toxic, ubiquitous and persistent chemical compounds, which can enter the food chain and accumulate up to higher trophic levels. Their determination requires sophisticated methods, expensive facilities and instruments, well-trained personnel and expensive chemical reagents. Ideally, real-time monitoring using rapid detection methods should be applied to detect possible contamination along the food chain in order to prevent human exposure. Sensor technology may be promising in this respect. This review gives the state of the art for detecting possible contamination with dioxins and DL-PCBs along the food chain of animal-source foods. The main detection methods applied ( In partortho PCBs share the planar structure and metabolic persistence of dioxins. Due to their toxicity and strict measures to reduce human exposure, dioxins and DL-PCBs are a major threat for production of safe feed and food [In addition to dioxins, polychlorinated biphenyls (PCBs) are a group of chemical compounds that have been produced for industrial purposes . Althougand food . Figure In addition to the DL-PCBs, there is the much larger group of non-dioxin-like PCBs. In practice only a limited number of these NDL-PCBs are determined as indicator compounds for the group . In most countries limits were set in the past for these so-called indicator PCBs. The EU decided to harmonize these limits and new limits will become effective in 2012. In general these limits are an order of magnitude higher than those for dioxins and DL-PCBs, also meaning that their detection will be easier to achieve. For milk e.g., the limit will be 40 ng/g fat as compared to 3 and 6 pg TEQ/g fat for dioxins and the sum of dioxins and DL-PCBs.et al. [i.e., validity, simplicity, sensitivity, relevance and feasibility. Finally, we will address the potential of biosensors for detection of dioxins and DL-PCB contamination throughout the food chain.Since the Belgian dioxin incident in 1999, which strongly affected consumer confidence in foodstuffs and caused huge financial losses , there het al. quantifiet al. . The golet al. . Such meet al. \u201317, enviet al. \u201321 and fet al. \u201325. Howe2.p-dioxin (TCDD) [in vivo experimental toxicity studies, each congener is assigned a relative potency factor or TEF, expressing the toxicity in comparison to TCDD (assigned TEF of 1). When analyzing a sample, the levels of individual congeners are multiplied by TEF values and summed to arrive at a TEQ-level [Among the different congeners of dioxins and DL-PCBs, the most toxic congener is 2,3,7,8-tetrachlorinated dibenzo-n (TCDD) . Other dn (TCDD) . In the n (TCDD) ,27. BaseEQ-level .TCDD is considered to be a human carcinogen . BesidesBased on the adverse effects in animals during animal experiments, the Scientific Committee on Food established a Tolerable Weekly Intake (TWI) level of 14 pg TEQ/kg bw/week. This limit should prevent that body levels in consumers will eventually reach a critical level. It was shown that part of the population still exceeds this TWI. The consumption of contaminated animal-source food appears the main source of dioxin and DL-PCB exposure in humans ,41. Theri.e., the higher numbers of chlorine molecule (7 or 8) congeners are transferred with lower carry-over rates than the lower chlorinated ones [In general, oral exposure is the main route of dioxin and PCB contamination for both animals and humans. After absorption via the gastrointestinal tract, some congeners are metabolized into non-toxic compounds and excreted ,43. In p 6) ones . Laying 6) ones .Since dioxins can enter the animal food chain in different ways, in this study, we applied a \u201ccradle-to-farm-gate\u201d approach to milk, egg and meat production in order to determine possible routes in which dioxins and DL-PCBs enter the food chain. Contamination due to post-farm processes, therefore, is not considered but based on the various incidents, this route seems less important. Dioxins and DL-PCBs mainly enter the food chain by oral ingestion of contaminated substances, such as compound feed or feed supplements, roughages, water, soil, and worms or insects. The level of exposure to these compounds in animal production systems is also affected by farm management. Nowadays, consumers pay more attention to animal production systems where food is produced in an animal-friendly way . An examDioxins can be potentially emitted from fires, including those from forest, bush and grassland, to the atmosphere \u201363. It hInterestingly, about 38% of dioxin exposure in the Dutch population can be attributed to the consumption of milk and milk products , primariThe presence of dioxins and DL-PCBs in milk mainly originates from the consumption of roughage since this is the main feed source for dairy cows . In addiApproximately 5% of total daily exposure of the Dutch population to dioxins and DL-PCBs has been estimated to originate from egg consumption . The incApplication of wood shavings and sawdust prepared from wood, which has been treated with pentachlorophenol (PCP) as animal bedding material, may also cause increased levels of dioxins in food products . It was Higher TEQ-values were observed in meat from outdoor than from indoor production systems . Ingestiet al. [The use of contaminated feed ingredients resulted in elevated levels of dioxins and PCBs in pork in Belgium , Chile , the Netet al. found thet al. .Monitoring programs for early detection of the contaminants in all production stages may be the most promising approach to limit dioxins and DL-PCB contamination in animal-source food. The use of critical control points . This method requires a sophisticated clean-up step to separate the compounds from fat and other contaminants that may interfere with the analysis. The HRGC-HRMS technique is at present the only accepted method that can quantify the concentration of different congeners of dioxins and DL-PCBs at very low detection limits . The insIn general, chemical analysis offers excellent sensitivity for measuring dioxin levels in food, because of its very low detection limits. The main drawback of chemical analysis based on HRGC-HRMS is the lack of information about potential other dioxin-like compounds, like the brominated or mixed halogenated dioxins . At pres3.2.Several bioassays, which commonly involve living organisms or tissues that sense toxic substances, have been established for detection and quantification of dioxin and PCB contamination in the food chain or its environment. These bioassays quantify the magnitude of contamination by the expression of responsive reporter genes. Gene expression is mediated by binding of ligands to the intracellular aryl hydrocarbon receptor (AhR) . The repAs the CALUX bioassay measures the response following ligand binding to the intracellular aryl hydrocarbon receptor, many other compounds including both natural and synthetic compounds can interfere with the assay. Superinduction of the aryl hydrocarbon receptor (AhR), caused by other AhR active substances such as polybrominated compounds, or activation of protein kinase C, resulted in an inaccurate overestimation of dioxins . Moreoveet al. [et al. [et al. [et al. [Several laboratories, therefore, have investigated methods to clean-up the sample thus improving the accuracy and precision of the CALUX method. Such a step is also required to separate the dioxins and DL-PCBs from the fat, which is essential for most types of samples. Hoogenboom et al. demonstr [et al. compared [et al. develope [et al. develope [et al. .et al. [et al. [Another method to improve the output from the CALUX bioassay has been recently reported by Zhao et al. . These a [et al. reportedRecently, Baston and Denison reportedOverall it is clear that cell-based bioassays like CALUX are very suitable for screening of feed and food, but also water and soil, for the presence of dioxins and DL-PCBs. It is also clear that the clean-up based on acid silica is an important step not only to remove the fat but also many of the non-dioxin-like AhR-agonists and as such increases the selectivity of the test.3.3.Sensor technology is nowadays gaining more interest as it is perceived to comprise several advantages over conventional chemical analysis and biological assays, for example, its simplicity, its cost-effectiveness and the possibility for real-time and on-site analysis. Physical and biological sensors (biosensors) are the two promising technologies that might be used for determination of dioxins and DL-PCBs throughout the food chain involving various types of specimens such as water, air, soil, feed, animal tissues and final products. In this review, the main focus lies on biosensors but we will also briefly discuss some physical sensors and combinations of both technologies.3.3.1.Even though this group of sensors has been successfully established and commercially applied for many industrial purposes, there is limited literature available that reports about the use of physical sensors for detecting dioxins and PCBs. Carbon nanotube technology offers the possibility for dioxin and PCB detection ,112. Thi3.3.2.Currently, biosensors are interesting because of their advantages such as rapid, on-line or on-site analysis, minimal waste production, low cost of energy, less use of chemical reagents, miniaturization and the et al. [The definition of biosensor has been given by Thevenot et al. as \u201ca seet al. .Theoretically, biosensors can be classified according to their biological recognition elements being used. These elements can be enzymes, antibodies, DNA, whole cells and other biological receptors. Some of these biorecognition elements have been studied for determining dioxins and PCBs.et al. [Immunosensors or antibody-based biosensors are very versatile . Antibodet al. successfet al. [Centi et al. used immet al. . The senet al. . It was et al. [Pseudomonas sp. P2 as biorecognition element based on optical detection. Technically, this microorganism can oxidize PCB molecules, which results in production of yellow meta ring-fission metabolites that can be measured through the absorption spectra by an optical transducer. This sensor can detect PCB contamination in extracts of soil samples. However, it failed to detect the contamination in non-extracted samples of soil [et al. was the presence of other yellow metabolites from unidentified factors which might hamper the accuracy and precision of the measurement.Whole cells or tissues can also be used as a sensing element in biosensors . The CALet al. develope of soil . In summet al. [A sensing element from this group was synthesized and designed to mimic a natural bioreceptor, such as antibody and enzyme , that caet al. have devet al. [Mascini et al. also useet al. . After set al. [Cytochrome c (Cyt c), a heme containing metalloprotein which is ubiquitious in a cellular context and involved in electron transfer processes, has been used as a biological recognition element to detect PCBs in aqueous solution by Hong et al. . A confo4.Validity: this criterion judges the potential of determination methods on dioxins and DL-PCBs in terms of accuracy and precision (high repeatability and conformity of measurements).Simplicity: this criterion focuses on the ease to use the selected determination method.Sensitivity: due to the ultra-low limits for dioxins and DL-PCBs in the food chain and related samples, highly sensitive recognition elements are needed to be able to qualify and/or quantify the contamination. This criterion is judged based on the potential of the detector for dioxins and DL-PCBs determination in terms of limits of detection.Relevance: this criterion is based on the interpretability of the output in relation to the biological toxicity.Economic and technical feasibility: this criterion is evaluated based on the cost-effectiveness and the possibility to use the method in a commercial context .The following criteria are generally used to assess the potential and limitations of sustainability indicators , and appBased on the above mentioned criteria, we assessed and scored each of the determination methods .Potential and limitations of HRGC-HRMS have been discussed elsewhere ,91. ThisThe major advantages of physical sensors are high sensitivity and specificity. However, the outputs do not show the level of biological toxicity and physical sensors are still in the developmental phase.Biosensors might be a promising technology for surveillance and monitoring the contamination of dioxins and DL-PCBs along the food chain because of their reliability, high throughputs and real-time determination. Timely, accurate and precise output can help decision-makers deciding the appropriate solutions for preventing contamination as an early warning system. These sensors are not yet applied in monitoring systems and still require further optimization and validation. Biosensors (in particular whole cell-based biosensors) usually reflect the biotoxicity of dioxins and DL-PCBs. The sensitivity of biosensors is lower compared with chemical analysis and bioassays. One of the advantages of biosensors is that they are primarily developed for ease of use. Within biosensor technology, whole cell-based biosensors present highest physiologically relevant output since they react to dioxins in a biologically relevant manner . A majorRegarding the very low limits for the samples, potential interference by other compounds and the variety of sample matrices, some pre-treatment methods are still needed prior to determination by biosensors and this is likely to be a bottleneck for their development . As a re5.This review provides insight into potential and limitations of different techniques for detecting presence of dioxins and DL-PCBs in the food chain of milk, eggs and meat. Since dioxins and DL-PCBs are highly toxic and ubiquitously present in food chain and its environment, efficient monitoring must be applied as an early warning system to prevent exposure of animals and humans.The need of real-time and on-site determination of dioxins is of utmost importance in order to make correct and timely decisions. Chemical analysis is the gold standard method, but requires expensive and sophisticated facilities and instruments, costly reagents and well-trained operators. The CALUX bioassay is an established method, that can efficiently be used for screening of samples, even though extraction and clean-up methods are a prerequisite. Sensors are promising for detection of dioxins and DL-PCBs in the food chain, although they are still under development. Different biorecognition elements provide different advantages and limitations. Overall, immunosensors and biomimetic-based biosensors present advantages, such as low variation between producing batches and specificity to individual congeners of dioxins and DL-PCBs. Limitations are the lack of biological output and the inability to detect other dioxin-like contaminants. The susceptibility of immunosensors to residues of organic solvents, which are normally used in the step of sample extraction and clean-up, may reduce their potential when compared to biomimetic-based biosensors. Meanwhile, the whole cell-based biosensors show a high potential for detecting contamination by providing useful biological output. Several limitations of whole cell-based biosensors e.g., sensitivity, selectivity, limits of detection and user-friendly aspects need to be further optimized. In addition to further optimization, biosensor methodology requires further standardization in order to allow their application for excluding contaminant levels in food and feed above existing limits."} +{"text": "Malignant tumours of minor salivary glands are uncommon, representing only 2-4% of all head and neck cancers. In the larynx, minor salivary gland tumours rarely occur and constitute less than 1% of laryngeal neoplasm. Most of the minor salivary gland tumours arise in the subglottis; however, they can also occur in the supraglottis, in the false vocal cords, aryepiglottic folds and caudal portion of the epiglottis. The most common type of malignant minor salivary gland tumour is adenoid cystic carcinoma.We present a unusual case of adenoid cystic carcinoma of glottic-subglottic region in a 61-year-old woman. Follow-up endoscopy and laryngeal magnetic resonance imaging (MRI) at three years after treatment showed no recurrence of the tumour.The diagnosis of glottic-subglottic adenoid cystic carcinoma should be considered in patients who are characterized by dyspnea, cough and stridor, but do not respond to pharmacologic approach.Adenoid cystic carcinoma is usually a very slow growing cancer, invested by an apparently normal laryngeal mucosa, so that it can show no clear symptoms for a long time. For these reasons the increasing number of diagnostic mistakes or late diagnosis that may be fatal in some cases. Adenoid cystic carcinoma is the predominant histologic type among malignancies of the minor salivary glands, with a frequency of 10-20%, representing only 2-4% of all head and neck malignancies ,4. Its pA.N, 61 year-old woman, with an ACC injury in the glottis-subglottis region, has been examined after a surgical treatment of thyroid. The clinical appearance of thyroid cancer is that of a nodules, some time representing a challenging diagnostic dilemma with thyroid or unusual extrathyroidal masses ,9. The p2+ toolkit, that has been observed in other malignancies, including renal cellular carcinoma [Aging is accompanied by a increased incidence of mortality from many diseases like cancer, diabetes, neurodegenerative, cardiovascular and other pathologies all related to oxidative stress and elevated ROS (Reactive oxygen species) ,11. Drugarcinoma ,25 and parcinoma , and hasarcinoma .Laryngeal salivary gland carcinomas are rare and account for < 1% of laryngeal malignancies. Therefore, a high degree of suspicion is essential for early diagnosis. This tumour must be considered when aggressive laryngeal tumours are found, especially if the patient is not at risk for squamous cell carcinomas. They usually originate in the supra-glottic or sub-glottic area with a predominance of old age. Most patients are diagnosed late, at an advanced stage. CT can be used to assess tumour extent and growth patterns. Wide-margin surgery alone or in combination with post-operative radiotherapy for advanced lesions that present peri-neural spread or close or positive margins is the best tumour management. For these patients, regular and long-term follow-up is mandatory, in order to detect relapses and metastases.The authors declare that they have no competing interests.DT: conceived the study, analyzed and interpreted the data, drafted the manuscript. GG: conceived the study, analyzed and interpreted the data, drafted the manuscript. GC: critically revised the manuscript. MN: critically revised the manuscript. GD: critically revised the manuscript. MS: analyzed the data. GI: analyzed the data. MV: critically revised the manuscript. FR: critically revised the manuscript. GM: conceived the study, critically revised the manuscript. All authors read and approved the final manuscript.DT: Assistant Professor of Otolaryngology at Second University of NaplesGG: Assistant Professor of Anatomy at University of MoliseGC: Assistant Professor of Surgery at Second University of NaplesMN: Resident in Otolaryngology Training Program at Second University of NaplesGD: Assistant of Otolaryngology at Second University of NaplesMS: Research Fellow at University of Naples \"Federico II\"GI: PhD Student at University of Naples \"Federico II\"MV: Assistant Professor of Endocrinology at University of SalernoFR: Assistant Professor of Dentistry at University of Naples \"Federico II\"GM: Full Professor of Otolaryngology at Second University of Naples"} +{"text": "Limited clinician involvement in smartphone application development poses problems considering the extensive use of smartphones among medical professionals and patients.We present a simple method for the clinician to develop simple web-apps using only an Internet browser and a text editor.This method may help clinicians develop simple web-apps and increase clinician involvement in smartphone content. The impressive distribution and use of smartphones among health professionals and patients promise interesting advances within fields such as medical education, patient education, patient aid, and telemedicine -3. HowevClinicians diligently develop digital and printed material, but may be reluctant from developing apps as required technical skills or resources may be insufficient. While some smartphone apps may require advanced algorithms or technology, many popular apps are actually quite simple and consist only of text and images. In fact, simple web-apps \u2014 smartphone applications that function using the build-in web browser \u2014 can be developed as easy as preparing slideshows. Here, we describe one easy method to develop a simple web-app by only using an Internet browser and a text editor.In general, the design consists of several different frames, of which only one is visible to the user. The user will start at the main frame, and then navigate to other frames. Each of these frames and the desired connections can be designed using the drag-and-drop user interface on the jQuery Mobile website based onTwo limitations of this method should be noted. Firstly, programming assistance may be needed for enabling more advanced functionalities. Secondly, web-apps are not indexed in iTunes App Store or Google Play, and therefore users will need a link to access the web-app, unless it is converted to native smartphone apps using conversion tools, such as PhoneGap or CodiqProject name: Designing web-apps for smartphones can be easy as making slideshow presentationsProject home page: None.Operating system(s): Platform independentProgramming language: HTML, CSS, and JSOther requirements: A simple text-editing program, e.g. NotepadAny restrictions to use by non-academics: None (MIT License)The data sets supporting the results of this article are included within the article and its additional files.Yousif Subhi, Tobias Todsen, Charlotte Ringsted, and Lars Konge have no conflicts of interests to declare.YS contributed with the original idea and drafted the manuscript. YS and TT recorded the instructional video. TT, CR, and LK helped to draft the final manuscript. All authors read and approved the final manuscript.YS is medical graduate student and TT is medical doctor at the University of Copenhagen and work at the Centre for Clinical Education at the University of Copenhagen and the Capital Region of Denmark investigating how smartphones are used in medical education. YS is also a research assistant at the Department of Ophthalmology at the Copenhagen University Hospital Roskilde. CR is director and scientist of the Wilson Centre and professor in the Department of Anesthesia at the University of Toronto. LK is clinical associate professor and senior consultant at the Centre for Clinical Education at the University of Copenhagen and the Capital Region of Denmark.Code-snip top. Description: Code-snip to be inserted at the top of the text-file.Click here for fileCode-snip bottom. Description: Code-snip to be inserted at the bottom of the text-file.Click here for fileInstructional video. Description: Instructional video on how to develop a simple web-app.Click here for fileExample web-app. Description: The example web-app developed in the instructional video.Click here for file"} +{"text": "Magnetic resonance imaging (MRI) was used to study the hand and wrist in very early rheumatoid arthritis (RA), and the results were compared with early and established disease.Fifty-seven patients fulfilling the new American College of Rheumatology criteria for RA, 26 with very early RA (VERA), 18 with early RA (ERA), and 13 with established RA (ESTRA), were enrolled in the study. MRI of the dominant hand and wrist was performed by using fat-suppressed T2-weighted and plain and contrast-enhanced T1-weighted sequences. Evaluation of bone marrow edema, synovitis, and bone erosions was performed with the OMERACT RA MRI scoring system.P < 0.05). No significant difference was found in synovitis.Edema, erosions, and synovitis were present in VERA, and the prevalence was 100%, 96.15%, and 92.3%, respectively. Significant differences in edema and erosions were found between VERA and ESTRA (Edema, erosions, and synovitis are findings of very early RA. MRI, by detecting these lesions, may play an important role in the management of these patients. Rheumatoid arthritis (RA) is a chronic systemic inflammatory disease characterized by prominent joint manifestations. Inflammation of the synovial membrane leads to the formation of a highly cellular inflammatory tissue, the pannus, which, by eroding cartilage and bone, leads to joint destruction and ankylosis . ArticulThe purpose of this study was to assess with MRI in very early RA (VERA) the prevalence and severity of hand and wrist involvement and to compare the involvement with early RA (ERA) and established RA (ESTRA).Fifty-seven consecutive unselected patients with RA, according to the new criteria for RA and withP value less than 0.05 was considered statistically significant.Statistical analysis was performed with SPSS base 15 for Windows. Interobserver variability was evaluated by using the Pearson correlation coefficient. Analysis of variance (ANOVA), followed by the least significant difference test, was used to study differences between groups. A P < 0.05 , grip strength (mm Hg), total joint count with tenderness or swelling, number of swollen joints, number of tender joints and pain score on VAS (cm)) and laboratory findings .In this study, the OMERACT RA MRI scoring system was applied to look for differences in bone edema, erosions, and synovitis between VERA, ERA, and ESTRA, and the major findings were (a) the presence of bone edema, erosions, and synovitis at the very early stages of RA; and (b) a significant difference in bone edema and erosions between VERA and ESTRA.MRI is being used largely in the assessment of hand and wrist involvement of patients with RA -7,13-16.et al [In this study, the presence of synovitis was independent of the disease duration, and the incidence was high in all groups. In the VERA group, the incidence of synovitis and bone erosions was almost similar. The exact nature of the relation between synovitis and bone damage remains unclear. The synovium seems to be the prime target in the inflammatory course of RA. Conaghan et al showed tet al -29.In the current study, a significant difference in edema and erosions was demonstrated between VERA and ESTRA. This is in agreement with previous studies, which, by using hand radiographs, showed that up to 60% of the patients develop joint erosions at the end of the 1 year from disease onset . The preIn conclusion, bone edema, erosions and synovitis are very early MRI findings of RA. MRI of the hand and wrist on clinical diagnosis of RA is useful to assess the degree of involvement.ANA: antinuclear antibodies; ANOVA: analysis of variance; CCP: citrullinated cyclic peptide; CRP: C-reactive protein; DAS-28: disease activity score of 28 joint indices; DMARDs: disease-modifying antirheumatic drugs; ERA: early rheumatoid arthritis; ESR: erythrocyte sedimentation rate; ESTRA: established rheumatoid arthritis; MCP: metacarpophalangeal; MRI: magnetic resonance imaging; RA: rheumatoid arthritis; RF: rheumatoid factor; TNF: tumor necrosis factor; VAS: visual analogue scale; VERA: very early rheumatoid arthritis.The authors declare that they have no competing interests.PEK helped to establish imaging techniques, analyze data, interpret data, and draft the manuscript. AKZ perform the examinations and analyzed data. MIA reviewed the interpreted data and critically reviewed the manuscript. PVV selected the patients and performed the musculoskeletal examination. AAD designed the study and critically reviewed the manuscript. All authors read and approved the final version of the manuscript."} +{"text": "APBioNet's mission, since its inception, has been to pioneer the growth and development of bioinformatics awareness, training, education, infrastructure, resources, and research among member countries and economies. Its work includes technical coordination, liaison, and/or affiliation with other international scientific bodies, such as the European Molecular Biology network (EMBnet) and the International Society for Computational Biology (ISCB). APBioNet has more than 20 organizational and 2,000 individual members from over 12 countries in the region, from industry, academia, research, government, investors, and international organizations. APBioNet is spearheading a number of key bioinformatics initiatives in collaboration with international organizations, such as the Asia-Pacific Advanced Network (APAN), the Association of South-East Asian Nations (ASEAN), the Asia-Pacific Economic Cooperation (APEC), and the Asia-Pacific International Molecular Biology Network (A-IMBN), and industry partners. Many of the initiatives and activities have been initiated through its flagship conference, the International Conference on Bioinformatics (InCoB). In 2012, APBioNet was incorporated in Singapore as a public limited liability company to ensure quality, sustainability, and continuity of its mission to advance bioinformatics across the region and beyond. We describe below the key thrust areas of APBioNet.The Asia-Pacific Bioinformatics Network . In 2011, InCoB celebrated its tenth anniversary in Kuala Lumpur, Malaysia jointly with the 1st ISCB-Asia http://incob.apbionet.org/incob11). InCoB2012 (http://incob.apbionet.org/incob12) marked the homecoming of the conference to its origin, Bangkok, as a reputable and major annual bioinformatics event in the Asia-Pacific region. The 2013 conference was held recently for the first time in China (http://incob.apbionet.org/incob13), with the 2014 conference to be held immediately prior to and sharing keynote speakers with the International Union for Pure and Applied Biophysics [IUPAB] 2014 Congress , in Sydney, Australia.InCoB is a conference series that started in Bangkok, Thailand in 2002. Since then, APBioNet has adopted InCoB as its annual signature event and grown it to become one of Asia's largest bioinformatics conferences, targeting practitioners from both biology and computing backgrounds (http://docid.apbionet.org); ii. reinstantiability and reproducibility (http://biodb100.apbionet.org); iii. author and contributor identity disambiguation (http://aid.apbionet.org); and iv. Minimum Information About a Bioinformatics investigation (MIABi) to include basic information necessary for an in silico experiment to be repeatable and the results reproducible, as well as harmonized with community initiatives in MIBBI http://biocurator.org), specifically BioDBcore (http://biocurator.org/biodbcore.shtml). We anticipate that the ongoing testing efforts In a multi-stakeholder effort to advance standards for bioinformatics activities, APBioNet has been working since InCoB2009 on building 100 exemplar biological and bioinformatics databases (BioDB100) and software tools (BioSW100) http://eabn.apbionet.org) held in Singapore (2008) that witnessed the proposal for minimum skills required of biologists in bioinformatics and biocomputation (msrBIC) www.apbionet.org/s-star) www.avist.org), to facilitate online/distance bioinformatics education and training. As a result of a research grant from the International Development and Research Centre (IDRC) of Canada and subsequent R&D at the National University of Singapore (NUS), APBioNet produced a bioinformatics grid-enabled software, as well as a distributable LiveOS containing bioinformatics software to facilitate bioinformatics training (http://en.wikipedia.org/wiki/BioSLAX). As part of its outreach, since 2003 APBioNet has partnered with various organizations to run bioinformatics workshops in the region (http://bit.ly/12eWyDe). In June 2012, APBioNet participated the inaugural Bioinformatics, Biotechnology, Biocuration and Computational Biology networks and societies (B3CB) meeting in Sweden that led to the establishment of the Global Organization for Bioinformatics Learning, Education and Training to coordinate bioinformatics training activities worldwide. All these efforts have contributed to the generation of skilled bioinformaticians in the region.APBioNet has been actively engaged in bridging the bioinformatics capability gap, with emphasis on establishing sustainable bioinformatics education and training www.apbionet.org/APAN/apan-apbionetMar98.html). This led to the formation of the BioMirrors project (www.bio-mirror.net) www.bic.nus.edu.sg/biogrid/biogrid01). More recently, APBioNet has been actively engaged in tracking the progress of advanced networking, such as the TransEurasia Information Network (TEIN2), developing cutting-edge initiatives, such as the International Workshop on World Wide Workflow Grid , and exploring applications of grid and cloud computing for life scientists . APBioNet is committed to continuing these efforts to meet the challenges of big data science in the decade ahead One of the earliest projects that APBioNet set up was a collaboration to build a bioinformatics network riding on the advanced network infrastructure of the Asia-Pacific Advanced Network (APAN) , the Philippines, Korea, Pakistan, Indonesia, Brunei, and many others Constructive scientific activism raises awareness among policy makers for the need to catalyze the transformation of life science research and education in the Asia-Pacific. APBioNet has worked closely with the ASEAN Committee on Science and Technology (COST) to assist in developing bioinformatics masterplans and roadmaps for the ten ASEAN member countries. Through the ASEAN Dialogue Partner mechanism, APBioNet is also actively engaging China, India, Japan, and Korea. Beyond these, APBioNet has also provided assistance to institutions in Pakistan and Saudi Arabia. As a result of these extensive, collective, and cooperative efforts to influence policy and raise awareness among policy makers and scientific leaders, Asian countries such as Singapore APBioNet has come a long way since its inception in 1998 at the Pacific Symposium for Bioinformatics, Hawaii. Currently, it is the largest regional bioinformatics organization in the Asia-Pacific, and one of the oldest. It continues to expand its presence in the region by actively reaching out to the research and education community, through its flagship conference InCoB. APBioNet is also taking steps toward setting standards and influencing scientific policy to enable a new generation of scientists to embrace the new biology of today that is increasingly information- and technology-driven, with knowledge generation dependent on applications of physical and computer sciences."} +{"text": "Identification of essential proteins is always a challenging task since it requires experimental approaches that are time-consuming and laborious. With the advances in high throughput technologies, a large number of protein-protein interactions are available, which have produced unprecedented opportunities for detecting proteins' essentialities from the network level. There have been a series of computational approaches proposed for predicting essential proteins based on network topologies. However, the network topology-based centrality measures are very sensitive to the robustness of network. Therefore, a new robust essential protein discovery method would be of great value.Saccharomyces cerevisiae. The experimental results show that the predicted precision of PeC clearly exceeds that of the other fifteen previously proposed centrality measures: Degree Centrality (DC), Betweenness Centrality (BC), Closeness Centrality (CC), Subgraph Centrality (SC), Eigenvector Centrality (EC), Information Centrality (IC), Bottle Neck (BN), Density of Maximum Neighborhood Component (DMNC), Local Average Connectivity-based method (LAC), Sum of ECC (SoECC), Range-Limited Centrality (RL), L-index (LI), Leader Rank (LR), Normalized \u03b1-Centrality (NC), and Moduland-Centrality (MC). Especially, the improvement of PeC over the classic centrality measures is more than 50% when predicting no more than 500 proteins.In this paper, we propose a new centrality measure, named PeC, based on the integration of protein-protein interaction and gene expression data. The performance of PeC is validated based on the protein-protein interaction network of We demonstrate that the integration of protein-protein interaction network and gene expression data can help improve the precision of predicting essential proteins. The new centrality measure, PeC, is an effective essential protein discovery method. The identification of essential proteins is crucial for understanding of the minimal requirements for cellular life , which iEssential proteins are those proteins necessary for growth in a rich medium where all the required nutrients are available . The delSaccharomyces cerevisiae, Caenorhabditis elegans, and Drosophila melanogaster [To break through these experimental constraints, some researchers have proposed various computational approaches. With the accumulation of data derived from experimental small-scale studies and high-throughput techniques, there is a growing awareness that the topological properties of biological networks would be useful for the identification of essential proteins. It has been observed in several species, such as nogaster ,13, thatnogaster . This isnogaster . Althougnogaster -18, mostnogaster -21.\u03b1-Centrality [Specifically, some global network characteristics, such as betweenness centrality and closntrality , and Modntrality , used inThough a great progress has been made on the computational methods for the identification of essential proteins based on network topologies, there are still several challenges that researchers have to meet. First, the protein-protein interaction dataset for each species is not complete up to now. Second, a high proportion of false positives has been found in protein-protein interaction networks, especially for those obtained by high-throughput technologies. In addition, as reported by Zotenko et al. , essentiSaccharomyces cerevisiae. Compared to other fifteen previous centrality measures: Degree Centrality (DC) [\u03b1-Centrality (NC) [With respect to these various difficulties and progresses, we propose a new centrality measure, named PeC, by integrating protein-protein interaction data and gene expression data. Different from other centrality measures, PeC determines a protein's essentiality not only based on its connectivity, but also whether it has a high probability to be co-clustered and co-expressed with its neighbors. The performance of PeC was tested on the well studied species of ity (DC) , Betweenity (DC) , Closeneity (DC) , Subgrapity (DC) , Eigenveity (DC) , Informaity (DC) , Bottle ity (DC) ,39, Density (DC) , Local Aity (DC) , Sum of ity (DC) , Range-Lity (DC) , L-Indexity (DC) , LeaderRity (DC) , Normaliity (NC) , and Modity (NC) , PeC achIn this study, a new centrality measure, PeC, is proposed based on the integration of protein-protein interaction data and gene expression data. The basic ideas behind PeC are as follows: (1) A highly connected protein is more likely to be essential than a low connected one; (2) Essential proteins tend to form densely connected clusters; (3) Essential proteins in the same cluster have a more chance to be co-expressed. In PeC, a protein's essentiality is determined by the number of the protein's neighbors and the probability that the protein is co-clustered and co-expressed with its neighbors.G, where a node v \u2208 V represents a protein and an edge e \u2208 E denotes an interaction between two proteins u and v. Gene expression is the process by which information from a gene is used in the synthesis of a functional gene product. These gene products are often proteins. Of course, there may exist some functional RNAs from non-protein coding genes. Here, we only consider the gene expressions for proteins. For a protein v, its gene expressions with s different times are denoted as Ge(v) = {g, g, ..., g}. The probability that two proteins are co-clustered and co-expressed is evaluated based on the edge clustering coefficient (ECC) and pearson correlation coefficient (PCC).To describe PeC simply and clearly, we provide the following definitions and descriptions. The protein-protein interaction network is represented by an undirected graph et al. [u, v) connecting node u and node v, we calculate its ECC by using the common neighbors instead of triangles. The ECC of an edge is defined as:Clustering coefficient was first proposed to describe the property of a vertex in a network, which has been used as an effective tool to analyze the topology of protein-protein interaction networks . Radicchet al. generaliet al. ,42, we het al. is not fu N(or vN) is the set of neighbors of vertex u (or v) and u d(or vd) denotes the degree of vertex u (or v), i.e., the number of nodes which u (or v) directly connects in graph G.where ECC is a local variable which characterizes the closeness of two proteins u and v. Obviously, two proteins u and v with a larger value of ECC are more likely to be in the same cluster.ECC is that it describes effectively the probability of two proteins being in a cluster from the topology view. However, it also has disadvantage. The effectiveness of ECC heavily depends on the reliability of the protein-protein interaction networks. Thus, in this paper we will introduce another metric, pearson correlation coefficient, which is independent of the reliability of the protein-protein interaction networks, to evaluate how likely two proteins are in the same cluster from another view.The advantage of X and Y), which encode the corresponding paired proteins (u and v) interacting in the protein-protein interaction network, is defined as:To evaluate how strong two interacting proteins are co-expressed, we calculate their pearson's correlation coefficient(PCC). The PCC of a pais is the number of samples of the gene expression data; g )) is the expression level of gene X (or Y) in the sample i under a specific condition; \u1e21(X) (or \u1e21 (Y)) represents the mean expression level of gene X (or Y) and \u03c3(X) (or \u03c3(Y)) represents the standard deviation of expression level of gene X (or Y). Here, we defined the pearson's correlation coefficient of a pair of proteins (u and v) as equal to the PCC of their corresponding paired genes (X and Y), that is PCC = PCC. The value of PCC ranges from -1 to 1. If PCC has a positive value, there is a positive linear correlation between u and v.where u and v to be in the same cluster as following:It has been proved that there exist a number of protein complexes which play a key role in carrying out biological functionality and the v, its PeC(v) is defined as the sum of the probabilities that the protein and its neighbors belong to a same cluster:For a protein v Ndenotes the set of all neighbors of node v.Where PeC(v) is determined by not only how many neighbors the protein has but also how likely it is co-clustered with its neighbors. In our previous studies [cp.The value of studies , we haveSaccharomyces cerevisiae, as it has been well characterized by knockout experiments and widely used in the evaluations of essential proteins. The test data used in this paper are as following:To evaluate the performance of the proposed new centrality measure, PeC, we implemented it on the discovery of essential proteins of Saccharomyces cerevisiae was downloaded from the DIP database [The protein-protein interactions of database . There aSaccharomyces cerevisiae were collected from the following databases: MIPS [A list of essential proteins of es: MIPS , SGD [48es: MIPS , and SGDes: MIPS . A proteSaccharomyces cerevisiae was retrieved from Tu et al., 2005 [The gene expression data of l., 2005 , containThe detailed information of proteins with gene expression data is shown in Additional file \u03b1-Centrality (NC) [To validate the performance of the proposed new centrality measure PeC, we carry out a comparison between it and fifteen other previously proposed centrality measures: Degree Centrality (DC) , Betweenity (NC) , and Modity (NC) .Proteins are ranked according to their values calculated by each centrality measure. A certain number of top proteins are selected as candidates for essential proteins. Then we determine how many of them are true essential proteins. The number of essential proteins detected by PeC and fifteen other centrality measures from the yeast protein-protein interaction network is shown in Figure From Figure A more general comparison between the proposed new centrality measure PeC and the fifteen previously proposed centrality measures is tested by using a jackknife methodology . The comAs shown in Additional file To further analyze why and how PeC performs well on the identification of essential proteins we study the relationship and difference between it and fifteen other centrality measures by predicting a small fraction of proteins. For each centrality measure, the top 100 proteins are selected. The information of the top 100 proteins of PeC and fifteen other centrality measures is shown in Additional file PeC \u2229 iM| denotes the number of common proteins detected by PeC and by a centrality measure iM, {i M- PeC} means the set of proteins identified by i Mnot by PeC, and |i M- PeC| is the number of proteins identified by i Mnot by PeC.Firstly, we compare PeC with DC, BC, CC, SC, EC, IC, BN, DMNC, LAC, SoECC, RL, LI, LR, NC, and MC by investigating how many proteins are both predicted by PeC and by anyone of the fifteen centrality measures. The number of overlaps between PeC and one of the other centrality measures is shown in Table From Table Secondly, we evaluate the different proteins identified by PeC and those by other centrality measures. Figure A list of proteins which are predicted by PeC but ignored by all the ten centrality measures when predicting the top 100 proteins are shown in Additional file Additional file Saccharomyces cerevisiae. The experimental results show that the predicted precision of PeC is clearly higher than those of the fifteen other topology-based centrality measures: Degree Centrality (DC), Betweenness Centrality (BC), Closeness Centrality (CC), Subgraph Centrality(SC), Eigenvector Centrality(EC), Information Centrality(IC), Bottle Neck (BN), Density of Maximum Neighborhood Component (DMNC), Local Average Connectivity-based method (LAC), Sum of ECC (SoECC), Range-Limited Centrality(RL), L-index(LI), Leader Rank(LR), Normalized \u03b1-Centrality(NC), and Moduland-Centrality(MC).The identification of essential proteins from the network level is a hot topic in the postgenome era. Many approaches based on topological characteristics have been proposed for predicting essential proteins in biological networks. Unfortunately, most of the topology-based methods depend on the reliability of the available protein-protein interactions and thus are very sensitive to the network. To overcome these difficulties, we propose a new centrality measure, named PeC, based on the integration of protein-protein interaction and gene expression data. PeC is applied to the protein-protein interaction network of Though PeC performs well on the identification of essential proteins, there may be still a space to improve the prediction performance. First, the integration of PCC and ECC is very simple in this paper. Further study on the relationship between PCC and ECC will provide new clues to integrating PCC and ECC in a more accurate way. Second, some other protein related data, such as biological process, domain information, and localization, besides the gene expression data, can also be integrated into the protein-protein interaction networks for identifying essential proteins. The integration of multiple protein related data may contribute a good deal to the identification of essential proteins with further research efforts.The authors declare that they have no competing interests.ML and HZ obtained the protein-protein interaction data, essential proteins and gene expression data. ML and HZ designed the new centrality, PeC, and analyzed the results. ML and HZ drafted the manuscript together. JW and YP participated in revising the draft. All authors have read and approved the manuscript.Information of the yeast protein-protein interaction network obtained from the DIP database. This file shows the number of proteins, essential proteins, non-essential proteins, and interactions of the yeast protein-protein interaction network obtained from the DIP database. (DOC 28 kb).Click here for filePeC is compared with fifteen recent centrality measures by a jackknife methodology. This file includes five figures: (a) PeC is compared with DC and DMNC; (b) PeC is compared with BC, SC and BN; (c) PeC is compared with CC, EC and IC; (d) PeC is compared with LAC and SoECC; (e) PeC is compared with RL, LI, LR, NC, and MC. To compare with the results of random sorting, ten random assortments are also plotted in each figure. The X-axis represents the ranked proteins in the yeast protein-protein interaction network, ranked from left to right as the highest to the lowest values of centrality measures. The Y-axis is the cumulative count of essential proteins with respect to the ranked proteins moving left to right. (DOC 7744 kb).Click here for fileThe top 100 proteins identified by PeC and other ten centrality measures. This file is composed by 11 groups of data corresponding to PeC and other ten centrality measures: Degree Centrality (DC), Betweenness Centrality (BC), Closeness Centrality (CC), Subgraph Centrality(SC), Eigenvector Centrality(EC), Information Centrality(IC), Bottle Neck (BN), Density of Maximum Neighborhood Component (DMNC), Local Average Connectivity-based method (LAC), Sum of ECC (SoECC). (XLS 36 kb).Click here for fileA list of 41 proteins predicted by PeC which are ignored by the ten centrality measures: DC, DMNC, BC, SC, BN, CC, EC, IC, LAC, SoECC when predicting the top 100 proteins. There are some proteins which are ignored by the ten centrality measures: DC, BC, CC, SC, EC, IC, BN, DMNC, LAC, and SoECC, but identified by PeC. This file provides the list of 41 proteins predicted by PeC which are ignored by all the ten centrality measures when predicting the top 100 proteins. (DOC 68 kb).Click here for fileA list of 25 non-essential proteins with a low value of PeC predicted by DC. The non-essential proteins predicted by DC which have a low value of PeC are shown in this file. For each non-essential protein, its values of SoECC, SoPCC, average of ECC, and average of PCC are also shown in this file. (XLS 17 kb).Click here for fileExamples of non-essential proteins which have high degree but with low PeC. Two examples of non-essential proteins YGR254W and YDL059C are shown. YGR254W and YDL059C both have a high degree of 67, but their PeC values are very low. The PeC value of YGR254W is 0.007 and that of YDL059C is -0.241. (DOC 246 kb).Click here for fileExamples of non-essential proteins which have high degree and high SoECC but with low PeC. Two examples of non-essential proteins YML048W and YHR140W are shown. YML048W and YHR140W not only have a high degree but also have a high value of SoECC. However, their PeC values are very low. The PeC of YML048W is -0.241 and that of YHR140W is -2.447. (DOC 518 kb).Click here for fileA list of 17 non-essential proteins with a low value of PeC predicted by SoECC. The non-essential proteins predicted by SoECC which have a low value of PeC are shown in this file. For each non-essential protein, its values of SoECC, SoPCC, average of ECC, and average of PCC are also shown in this file. (XLS 24 kb).Click here for file"} +{"text": "The clinical benefits of repetitive transcranial magnetic stimulation (rTMS) for Parkinson's disease (PD) remain controversial. We performed a comprehensive study to examine whether rTMS is a safe and effective treatment for PD. Twelve PD patients received rTMS once a week. The crossover study design consisted of 4-week sham rTMS followed by 4-week real rTMS. The Unified Parkinson's Disease Rating Scale (UPDRS), Modified Hoehn and Yahr Stage, Schwab and England ADL Scale, Actigraph, Mini-Mental State Examination, Hamilton Depression Scale, Wechsler Adult Intelligence Scale-revised, and cerebral blood flow (CBF) and cerebrospinal fluid (CSF) examinations were used to evaluate the rTMS effects. Under both drug-on and drug-off conditions, the real rTMS improved the UPDRS scores significantly, while the sham rTMS did not. There were no significant changes in the results of the neuropsychological tests, CBF and CSF. rTMS seems to be a safe and effective therapeutic option for PD patients, especially in a wearing-off state. Parkinson's disease (PD) is the second most common neurodegenerative disease after Alzheimer's disease. The prevalence of PD in Japan has been estimated to be about 100 per 100,000 population \u20135. The tTwelve PD patients (seven men and five women) with a mean age of 69.2 years (range: 57\u201378 years) were included in this study . We usedTMS was performed with an SNM-1100 magnetic stimulator with a YM-121B large round coil . The stimulus parameters and the treatment protocol in the present study were according to the study of Shimamoto and Shigemori , which sFor the quantitative estimation of the outcome of rTMS treatments, the following clinical test batteries were used. The motor function and severity of PD were estimated by the Unified Parkinson's Disease Rating Scale (UPDRS), Modified Hoehn and Yahr Stage, Schwab and England ADL Scale, and Actigraph. For the estimation of the psychiatric effects of rTMS, the Mini-Mental State Examination (MMSE), Hamilton Depression Scale (HAM-D), and Wechsler Adult Intelligence Scale-revised (WAIS-R) were used. For the estimation of changes in cerebral blood flow (CBF) and possible harmful effects on the brain, single photon emission computed tomography (SPECT) and cerebrospinal fluid (CSF) examinations were conducted, respectively.Board-certified neurologists blindly estimated the motor function and severity of the parkinsonian symptoms. Without having any information of the patients on the drug-on or drug-off and the conditions of real or sham rTMS, the neurologists viewed a video record of each patient under six conditions: before sham rTMS (drug-on and drug-off), after sham rTMS (drug-on and drug-off), and after real rTMS (drug-on and drug-off). Under the drug-on condition, patients took antiparkinsonian drugs as prescribed. Under the drug-off condition, the morning doses of L-dopa compounds, bromocriptine, amantadine, and trihexiphenidyl were aborted on the examination day. The doses of pergolide were stopped from the night of the day before the examination day. The doses of cabergoline and selegiline were stopped from the morning of the day before the examination day. Patients took the prescribed antiparkinsonian drugs immediately after the drug-off examinations. UPDRS was scored at six states: before treatment (drug-on and drug-off), after sham rTMS treatment (drug-on and drug-off), and after real rTMS treatment (drug-on and drug-off). The Modified Hoehn and Yahr Stage was used to estimate the severity of PD patients. Stage 0 indicates no symptoms, and Stage 5, the worst state. The Schwab and England ADL Scale is a scale of activity of daily life, and it is scored from 0 to 100% according to the independence of voluntary actions in daily life. The Modified Hoehn and Yahr Stage and Schwab and England ADL Scale were scored under the same six conditions as for UPDRS scoring. An Actigraph is a wrist watch-like electronic device that records the frequency of arm movements , 17. TheThe effects on the patients' psychiatric conditions were examined by the MMSE, WAIS-R, and HAM-D. The MMSE is a question battery consisting of orientation, registration of words, attention, calculation, recalling of words, language, and visual construction . Intelli133Xe was used as a radionuclide. CBF was measured under three conditions: before treatment, after sham rTMS treatment, and after real rTMS treatment.CBF was estimated by brain SPECT. CSF examinations were performed twice: before sham rTMS treatment and after real rTMS treatment. Total protein, monoamine metabolites (homovanillic acid (HVA), 5-hydroxy indoleacetic acid (5-HIAA), 3-methoxy-4-hydroxyphenylethylene glycol (MHPG)), and neuron-specific enolase (NSE) in CSF were measured. The total protein concentration was determined by the pyrogallol-red method. The concentrations of HVA, 5-HIAA, and MHPG were determined by high-performance liquid chromatography (HPLC) . NSE wast-test with Bonferroni correction was used for post hoc analysis of the results of UPDRS. The one-way ANOVA was used to analyze the results of WAIS-R. The paired t-test was used to analyze the results of CSF examinations. Statistical significance was accepted at P < 0.05.Statistical analysis was performed with the SPSS 11.0 statistical analysis package . The repeated measures ANOVA were used to analyze the results of the UPDRS, Modified Hoehn and Yahr Stage, Schwab and England ADL Scale, Actigraph, MMSE, HAM-D, and CBF examinations. The paired Subjective improvements of resting tremor (five patients), painful dystonia of the legs (one patient), voice loudness (three patients), bending posture (one patient), wearing-off phenomenon (one patient), and akinesia (one patient) were observed after the rTMS treatments, and these lasted for one to three months. P = 0.006, repeated measures ANOVA). The real rTMS treatment improved the UPDRS scores significantly , while the sham rTMS treatment did not , 43.58 \u00b1 12.33, and 40.15 \u00b1 11.99, respectively . Throughrection) . The Mods ANOVA) . Schwab s ANOVA) .P = 0.003, repeated measures ANOVA). The real rTMS treatment improved the UPDRS scores significantly , while the sham rTMS treatment did not . The Mods ANOVA) . The Schs ANOVA) .P = 0.114, repeated measures ANOVA) (The results of the Actigraph examination (mean count per one minute and percentage of time of resting) while the subjects were awake or asleep did not change significantly among the three conditions . The MMSs ANOVA) . The HAMs ANOVA) . The scos ANOVA) . In the s ANOVA) . In the s ANOVA) . In the s ANOVA) .In the present study, the real rTMS treatments significantly improved the scores of UPDRS in PD; however, the sham rTMS treatments did not, indicating that rTMS may be effective for PD symptoms. In the clinical settings, it is a noteworthy finding that the rTMS treatment had a therapeutic effect under the drug-off condition, as it is analogous to the wearing-off phenomenon. In the advanced stage of PD, patients suffer from the instability of drug effects, such as the wearing-off or on-off phenomenon . The preThe scores of Actigraph, Schwab and England ADL Scale, and Modified Hoehn and Yahr Stage were not significantly changed before and after rTMS. Since the degree of improvements of PD symptoms by rTMS was small in the present study, it seems that only the UPDRS was able to detect this small difference of the symptomatic improvement of PD. In other words, for the functional evaluation of PD patients, the UPDRS seems to be the most sensitive among the functional measures used. The present study has several advantages over previous studies using rTMS for PD. Each study was different in terms of the coils used and the sites, intensities, frequencies, and total number of stimulations. Accordingly, the results cannot be compared equally. However, the present study had clear methodological advantages over previous studies. Firstly, we applied both the sham and real rTMS treatments in a crossover design. Secondly, the evaluations were performed under both drug-on and drug-off conditions. Thirdly, we estimated motor functions by the UPDRS score in a blind manner. Lastly, we performed a comprehensive study including the neuropsychological, CBF, and CSF examinations as well as the motor function of PD.In the present study, no adverse effects of the rTMS were observed. The MMSE scores slightly improved through the three trials, but the changes were not statistically significant. In the WAIS-R tests, the verbal IQ and performance IQ showed a slight improvement but without a statistical significance. The possibility cannot be excluded that, although not significant, the slight improvement of the results of the WAIS-R tests may be due to a learning effect. In any case, the results seem to indicate that the rTMS treatment may not have any significant beneficial or adverse effect on mental and psychiatric conditions. In addition, the rTMS treatment in our protocol did not appear to cause any damage in the central nervous system of the patients because the CBF and CSF concentrations of total protein and NSE did not change before and after rTMS. NSE is known to increase in CSF when neurons are injured rapidly, as in Creutzfeldt-Jakob disease and in the acute stage of cerebral infarction , 24. The mechanism of the therapeutic effect of rTMS for PD has not been clarified. Dopaminergic antiparkinsonian drugs have been reported to increase the CSF concentration of the monoamine metabolites, including HVA . In the One possible explanation for the therapeutic effects of the rTMS on PD symptoms is as follows. In PD, the excitability of the cerebral cortex is suspected to be decreased because of the altered excitatory inputs from the thalamus; TMS may compensate for this decreased excitability of the cerebral cortex \u201328. AnotIn conclusion, the present study demonstrates that rTMS seems to be a safe and effective therapeutic option for PD symptoms, especially in the wearing-off state."} +{"text": "Latimeria menadoensis is a coelacanth species first identified in 1997 in Indonesia, at 10,000 Km of distance from its African congener. To date, only six specimens have been caught and just a very limited molecular data is available. In the present work we describe the de novo transcriptome assembly obtained from liver and testis samples collected from the fifth specimen ever caught of this species.de novo assembled using a Trinity/CLC combined strategy. The assembly output was processed and filtered producing a set of 66,308 contigs, whose quality was thoroughly assessed. The comparison with the recently sequenced genome of the African congener Latimeria chalumnae and with the available genomic resources of other vertebrates revealed a good reconstruction of full length transcripts and a high coverage of the predicted full coelacanth transcriptome.The deep RNA sequencing performed with Illumina technologies generated 145,435,156 paired-end reads, accounting for ~14\u00a0GB of sequence data, which were The RNA-seq analysis revealed remarkable differences in the expression profiles between the two tissues, allowing the identification of liver- and testis-specific transcripts which may play a fundamental role in important biological processes carried out by these two organs.de novo transcriptome assembly can be considered a valuable support tool for the improvement of gene prediction within the genome of L. chalumnae and a valuable resource for investigation of many aspects of tetrapod evolution.Given the high genomic affinity between the two coelacanth species, the here described One of the most important transitions in vertebrate evolution was the arising of terrestrial vertebrates, which entailed considerable morphological changes related to the acquisition of novel functions by pre-existing and, in several cases, pre-adapted structures, like the evolution of lobe fins into tetrapod limbs. The terrestrial vertebrates would have derived from fossil forms of lobe-finned fishes, a highly successful group in the Devonian (400 Mya), with hundred species populating the Gondwana supercontinent\u2019s oceans and river systems ,2.Latimeria chalumnae), a fish considered extinct, was found off the estuary of river Chalumna, in South Africa The authors declare that they have no competing interests.EO, AC, AP and GS planned and coordinated the project. DMM collected the tissue samples. MF, MAB, MB, AC and EO prepared and sequenced the liver and testis RNA-seq libraries and performed the transposable elements analysis. AP, MG, GDM and JT-M performed the transcriptome assembly and annotation. AP, MG, GDM, AC, MF, MAB, MB, EO, GS, FB and AMF performed the transcriptome analyses. JA, FDP and JT-M compared the transcriptome data with the African coelacanth genome. AP, AC, MG, GDM and MF wrote the paper with input from other authors. All authors read and approved the final manuscript.a: Distribution of average sequence quality scores. The quality score for each read is calculated as the arithmetic mean of its base qualities. PHRED score is represented on the x-axis, the proportion of sequences observed at each score is shown on the y-axis. b: Coverage for the four nucleotides and ambiguous bases. The base position relative to each read is indicated on the x-axis, the percentage of each nucleotide observed at a certain position is shown on the y-axis. c: Combined coverage of G and C bases. The base position is shown on the x-axis, the percentage of G and C bases observed at each position is shown on the y-axis. d: Combined coverage of ambiguous bases. The base position is shown on the x-axis, the percentage of ambiguous bases observed at each position is shown on the y-axis. e: Ortholog Hit Ratio, calculated on the high quality set of liver and testis transcripts. The ratio of length between assembled contigs and the full length orthologs is reported on the x-axis, the percentage of contigs observed in each ratio category is shown on the y-axis. f: Gene Ontology mapping performed on the high quality transcript set. The mapping summary takes into account annotations at Level 2 of Cell Component, Molecular Function and Biological Process. Supplementary Methods: transcriptome assembly; transcripts integrity evaluation; comparison between the two coelacanth species.Click here for file"} +{"text": "Pushpa Verma's name was inadvertently omitted in the list of author list for the article. The authors would like to revise the author list to read as below:Jan-Michael Kugler, Pushpa Verma, Ya-Wen Chen, Ruifen Weng, Stephen M.CohenPushpa Verma 's affiliation is Institute of Molecular and Cell Biology, Singapore."} +{"text": "DC was supported by a grant of the Romanian National Authority for Scientific Research, CNCS \u2013 UEFISCDI, project number PN-II-ID-PCE-2011-3-0173. VDP was partly supported by a David H. Smith Conservation Research Fellowship from the Society for Conservation Biology (www.smithfellows.org). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\"A grant from the Romanian National Authority for Scientific Research to the third author was incorrectly omitted from the Funding Statement. The Funding Statement should read: \"This work was supported by a grant of the Romanian National Authority for Scientific Research, CNCS \u2013 UEFISCDI Diversity and distribution of reptiles in Romania. Zookeys 341: 49-76. doi: 10.3897/zookeys.341.5502."} +{"text": "There was an error in the funding statement. The correct funding statement is: \"This work is supported in part by the National Science Council, Taiwan, under Contract NSC100-2319-B-010-002, and in part by the \"A grant from Ministry of Education, Aim for the Top University Plan\" of the National Yang-Ming University, Taiwan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "International Code of Botanical Nomenclature to allow electronic-only publishing of new taxa. Even before the end of the Congress and formal acceptance of the changes PhytoKeys was able to publish a report on the main outcomes of the Nomenclature Section on electronic publishing the appearance of electronic publications as amendments or even alternatives to paper publications; (2) Open Access (OA) as a new publishing model; (3) the linkage of electronic registers, indices, and aggregators, which summarize information on biological species through taxonomic names or their persistent identifiers; and (4) Web 2.0 technologies, which permit the semantic markup of, and semantic enhancements to, published biological texts. The appearance of the journal was concomitant with lively discussions on the validity of nomenclatural acts published electronically . At the blishing .pre-publication registration of nomenclatural acts. Although the Nomenclature Section in Melbourne declined a proposal for mandatory pre-publication registration for acts in plants and algae, it approved the mandatory registration of fungal names on and after 1 January 2013 identifiers in all original descriptions of new species (protologues) and hence a workflow for species .Encyclopedia of Life, the Plazi Treatment Repository, and Wiki (Species-ID) on the day of publication. PhytoKeys also provides an established infrastructure for data publishing in cooperation with the Global Biodiversity Information Facility (GBIF) and the Dryad Data Repository.PhytoKeys has also been at the vanguard of \u201catomized\u201d content, i.e., to separately distribute the taxonomic information included in a paper to relevant on-line aggregators. Thanks to its advanced XML-based editorial workflow, the journal exports taxon treatments to the www.sourceforge.net/projects/taxpub), an extension of the Journal Archiving and Interchange Tag Suite (JATS) maintained by the U.S. National Center for Biotechnology Information (NCBI) of the U.S. National Library of Medicine (NLM), all papers published in PhytoKeys will be archived in PubMedCentral in three versions, as PDF International Code of Nomenclature for algae, fungi, and plants . Authors are not requested to provide registration numbers, as the whole process of registration is provided by the Editorial Office of the journal in collaboration with IPNI\u2022\tPublish each paper in four versions: (1) PDF for effective publication, reference and easy archiving; (2) full-colour, high-resolution print version identical to the effectively published PDF version; (3) HTML for easy reading, browsing and applying semantic enhancements to the text; and (4) XML to provide a machine-readable file for archiving and data mining\u2022\tProduce a print version, identical to the PDF, which will be deposited it in six important botanical libraries of the world: Smithsonian Institution, Washington D.C.; Natural History Museum, London; Royal Botanic Gardens, Kew; Missouri Botanical Garden, St. Louis; Komarov Botanical Institute, St. Petersburg; Kunming Institute of Botany Heilongtan, Kunming, China.\u2022\tShorten the publication time to a maximum of one to two weeks after the editorial acceptance of a manuscript\u2022\tContinuously develop and implement cutting-edge publishing technologies: XML-based editorial work flow and mark up process, data publication and various semantic Web 2.0 enhancements.Plazi, the Global Biodiversity Information Facility (GBIF), the Encyclopedia of Life (EOL), the Biodiversity Heritage Library (BHL), the National Library of Medicine of the U.S. (NLM), and the ViBRANT EU FP7 project. We also PageBreakthank the staff of IPNI for helping us to establish a workflow for the provision of IPNI identifiers for new species of flowering plants.Finally, we would like to thank all of the authors, editors and readers of PhytoKeys for their support of the journal, as well as the translators of the paper of"} +{"text": "More than one in five people admitted to an ICU will die there. Research has highlighted concerns about support for patients and families and decision-making in this context ,2. Here,Medical Research Council guidance for complex interventions Phase 0 to I comprised literature review, theoretical modelling, observation and qualitative interviews and focus groups with staffand families exploring concerns and views of interventions identified in the literature review. Phase II comprised intervention development, implementation and evaluation of tool feasibility and effects using staff survey, observation, audit of records and relative survey.Phase I: 47 staffand 24 family members were interviewed. The short time between decisions for treatment withdrawal and death, plus concerns for support management, communication and decision-making, highlighted a need to ensure excellent psychosocial assessment for all. Phase II: as part of integrated care guidelines, we developed the King's Psychosocial Assessment and Care tool (K-PACE). K-PACE is used for all patients entering the ICU, completed within 24 hours of admission. It contains psychosocial assessment of the family and patient needs, and identifies key individuals for contact. Educational training was supported by K-PACE and was implemented in two waves. Post-implementation survey of 95 ICU stafffound that most (80%) were aware of K-PACE. Eighty-two per cent of nurses but only 17% of doctors had completed the tool. In total, 158/213 (74%) family members responded to the survey . There were high levels of satisfaction for symptom control and psychosocial care but concerns continued regarding explanation of treatment and care.K-PACE is a feasible tool to improve the palliative care of patients and their families in the ICU. Further refinement is needed and planned, with consideration of roll-out into the wider medical centre."} +{"text": "Encoding of stimuli in the retina depends on the statistical properties of the input stimuli, neural noise, and circuit nonlinearities. Here, we present a simple model of a two-path ON/OFF RGC circuit figure . We use"} +{"text": "Global Mental Health (GMH) in a spirit of celebration grounded in deep awareness of responsibility. In concert with the Journal's mission of moving from making the case to implementing GMH section of Belkin, , the T&L Belkin, . This pr\u2022Innovations in the development of feasible, contextually informed, effective, and cost-effective training with members of the mental healthcare workforce, both specialists and non-specialists.\u2022Provider and supervisor skill competency assessment, life-long learning, and skill reinforcement programs, ongoing program quality monitoring and improvement procedures, provider burden reduction and burnout prevention, user-friendly clinical management and decision-support m- and e-tools, organizational/policy factors supporting ongoing skill development, wellbeing of trainers and trainees, etc.\u2022Increase of community engagement and advocacy, family and person involvement in treatment, community mental health literacy, stigma reduction strategies and community impact of interventions on mental health knowledge and attitudes, etc.\u2022Effective strategies to maximize the reach of state-of-the-art mental health skills to academics, practitioners, and trainees globally; ethics in training; models for sustainable capacity-building and brain-drain prevention strategies; development of new models of learning communities, collaboratives, and global classrooms; managing learning stimulus \u2018overload\u2019 in the e-age, etc.With teaching and learning of the mental health workforce as its central axis, the T&L priority areas include:We hope that the T&L platform will set in motion a number of dialogues, discovery paths, and collaborations stemming from research projects, training programs, services, and policy/advocacy initiatives. Thus, teachers and learners in this section can not only be clinicians, and researchers, but also primary care personnel, development/aid organization workers, managers, policy makers, religious leaders, community members, and most critically, families and patients themselves. The common denominator is a focus on under-resourced systems in places where communities struggle with chronic adversity, toxic stress, and social exclusion on the one hand, and low availability of and access to mental health care for those who need it on the other. The problems that GMH tackles are everybody's problems.et al.task-shifting strategies (or task-sharing when there is an available team to share the tasks) . It will also hopefully see growth in learning-driven implementation strategies, such as Quality Improvement, that empower local decision-makers and stakeholders to be more effective implementers.In its most dynamic and comprehensive initiative to date, the WHO Mental Health and Substance Abuse department launched the mhGAP Intervention Guide (World Health Organization, Although a lot of attention is historically given to the quality of evidence that informs the selection of elements of care \u2013 the selection of a specific psychotherapy for example \u2013 there is significantly less on the quality of evidence of the training tools and processes themselves: did the training workshop increase knowledge in the domains targeted? Was there exploration of the cultural relevance of the training material? Was there a systematic process for its contextual adaptation? One point, however, of broad agreement, at least in theory, is the critical role of supervision for skill-building. There is a lot to be learned about facilitating and \u2018culture-changing\u2019 factors on the system and policy level, so that protected time for supervision becomes part of the workload and supervisors are not merely \u2018compliance monitors\u2019 but rather sources of support, knowledge, and improvement agents of providers' quality of work and life. The field is also looking for new ways of providing easily accessible stakeholder-driven feedback on training tools and methods. Finally, sustainable and scalable ways of capacity-building within academic institutions, which prepare the next generation of trainers, supervisors, and providers, as well as policies to prevent brain drain, are areas of great relevance to T&L. These topics are relevant to high- and low-income areas alike, and while we look to highlight solutions for low-resourced settings, we also look for contributions from any setting that inform us on shared challenges of broad applicability and interest.GMH training poses increasing and novel demands. To meaningfully engage and share knowledge with adult learners from vastly different professional, cultural, geographical, and economic backgrounds, we need to be \u2018multilingual\u2019 discipline-wise. We need guidance from fields like adult education and learning, information management sciences, public health, social sciences, and therapeutics, amongst others.et al.A learning domain we cover in this section involves the task-shifting aspects of care to the person and family, usually termed patient and family engagement and psychoeducation. In recent years, most gains in the management of chronic diseases were achieved by self-monitoring (Pearson globalmentalhealth@cambridge.org.The demands of distance-learning and the need for training large numbers of learners are giving technology a leading role. In addition to geography, other concerns such as availability of experts, safety, health, climate, gender-related concerns, time commitments, cost, etc. make internet-based learning a realistic educational option for a large number of learners. Mental health internet-based models offer scalable alternatives to widely used in-person cascade models of \u2018training-of-trainers\u2019 but also increase access to specialized training when needed. The development and testing of such a model on a large scale is expected to greatly inform the field (Fairburn & Patel,"} +{"text": "Three authors are not included in the author byline.th author and affiliated with the Laboratory of Translational Genomics, Division of Cancer Epidemiology and Genetics, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. The contributions of this author are as follows: Performed the experiments, contributed reagents/materials/analysis/tools.Laufey T. Amundadottir should be listed as the 8th author and affiliated with the National Cancer Institute, Division of Cancer Epidemiology & Genetics, Rockville. MD. The contributions of this author are as follows: performed the experiments, funded GWAS.Stephen J Chanock should be listed as the 15th author and affiliated with the Kaiser Permanente Division of Research. 2000 Broadway, Oakland, CA 94612. The contributions of this author are as follows: conceived and designed the experiments, wrote the manuscript.Stephen Van Den Eeden should be listed as the 44"} +{"text": "Here we present a summary of the 2014 International Conference on Intelligent Biology and Medicine (ICIBM 2014) and the editorial report of the supplement to BMC Genomics and BMC Systems Biology that includes 20 research articles selected from ICIBM 2014. The conference was held on December 4-6, 2014 at San Antonio, Texas, USA, and included six scientific sessions, four tutorials, four keynote presentations, nine highlight talks, and a poster session that covered cutting-edge research in bioinformatics, systems biology, and computational medicine. The 2014 International Conference on Intelligent Biology and Medicine (ICIBM 2014) was held on December 4-6, 2014 in downtown San Antonio, Texas, a popular city with the world famous River Walk filled with live music. More than one hundred researchers with diverse background spanning biology, medicine, computer science and engineering, statistics, and mathematics, among others attended the three-day event. The event promoted all attendees to exchange ideas, showcase recent innovative work, and foster interdisciplinary and multidisciplinary research collaborations, and provided education and training opportunities to students and junior investigators in bioinformatics, systems biology, intelligent computing, and computational medicine.The scientific program included six scientific sessions, four tutorials, four keynote presentations, nine highlight talks, and a poster session. The details of all presentations are available on the conference website and in tFour keynote speakers who are world-renowned leaders in bioinformatics, genomics, systems biology and computational medicine delivered lectures on their cutting-edge research and shared their views and perspectives of their research fields. These speakers were Dr. Tim Huang from University of Texas Health Science Center at San Antonio, Dr. Josh Stuart from University of California, Santa Cruz, Dr. Lynda Chin from University of Texas MD Anderson Cancer Center, and Dr. Jasmine Zhou from University of Southern California.\"Single-Cell Analysis of Tumor Heterogeneity\" Intratumor heterogeneity is a critical impediment to improving prognosis of many types of cancer such as breast and prostate cancer. In his lecture, Dr. Tim Huang presented a quantitative model to identify subpopulations of prostate single cells extracted from urine. Using a binary-coding system to identify unique concordant expression patterns among genes, the model resulted in a digital rendering of single-cell gene expression which enables non-invasive prognosis of prostate cancer patients. Dr. Tim Huang is Alice P. McDermott President's Distinguished Professor and Professor and Chair of the Department of Molecular Medicine at the University of Texas Health Science Center, San Antonio. He is also the holder of Max and Minnie Tomerlin Voelcker Distinguished University Professor and Deputy Director of the Cancer Therapy and Research Center, San Antonio. Dr. Huang is a Fellow of the American Association for the Advancement of Science.\"Identifying All of Cancer's Manifestations through Integrated Pan Cancer Analysis\" Dr. Josh Stuart presented the recent work of The Pan-Cancer Initiative of The Cancer Genome Atlas (TCGA) Research Network, including the analysis of thousands of human tumors to discover molecular aberrations at the DNA, RNA, protein and epigenetic levels in order to uncover data-driven tumor subtypes. Dr. Stuart described an integrated picture of commonalities, differences and themes across tumor lineages emerging from this large-scale, comprehensive work. While cancers are primarily classified on the basis of the body location where the disease originates, according to their new study, however, one in ten cancer patients would be classified differently using a new classification system based on molecular subtypes instead of the current tissue-of-origin system. Analysis of the molecular aberrations and their functional roles across tumor types will teach us how to extend therapeutics effective in one cancer type to others with a matching genomic profile, and to construct personalized networks for use in developing combinatorial therapy. Dr. Josh Stuart is Professor of Biomolecular Engineering and Associate Director of Center Biomolecular Science and Engineering at the University of California, Santa Cruz. He co-leads a Genome Data Analysis Center for the TCGA project, co-chairs the pan-cancer TCGA effort, is a leader of the bioinformatics pathways group for the International Cancer Genome Consortium (ICGC), and directs the computational pathway analysis for a Stand Up To Cancer (SU2C) Dream Team to identify therapies for resistant prostate cancer. He is an Alfred P. Sloan Fellow, and received an NSF CAREER award in 2009.\"Genomic Medicine: Transforming Cancer Research and Care\" Dr. Lynda Chin described a new model of integration, collaboration and cooperation between the research and clinical care enterprises, and between academia and industry, to bring to bear the power of technology and patient data on the cancer problem. She discussed the MD Anderson Cancer Center's APOLLO-Big Data platform and a prototype consumer-centric Amazon-like care delivery ecosystem in partnership with major industry giants, including IBM-Watson, to build MD Anderson Oncology Expert Advisor\u2122cognitive decision support system. Dr. Lynda Chin is Professor and Chair in the Department of Genomic Medicine, The University of Texas MD Anderson Cancer Center. For the 13 years prior to joining the University of Texas MD Anderson Cancer Center in 2011, Dr. Chin had been a professor of dermatology at Dana-Farber Cancer Institute and Harvard Medical School and a senior associate member of the Broad Institute of MIT and Harvard, and the Scientific Director of the Belfer Institute for Applied Cancer Science. Dr. Chin is the founding chair of the Department of Genomic Medicine at MD Anderson Cancer Center. She also serves as the scientific director of the Institute for Applied Cancer Science, which merges the best attributes of academia and industry to enable science-driven drug discovery. Dr. Chin was elected a member of the Institute of Medicine of the National Academies in 2012.\"Make Big Data Useful: Horizontal and Vertical Data Integration to Study Genes, Networks and Diseases\" Dr. Jasmine Zhou presented several interesting machine learning and graph algorithms for integrating data of the same type (e.g. gene expression), and of different types of data, such as epigenetic data, gene expression, and genome structures, in order to answer novel biological questions. She presented her recent work in integrating many RNA-seq datasets to perform high-resolution functional annotation of human genome, namely, predicting the functions of individual transcript isoforms, transforming the public gene expression repositories into a disease diagnosis database, and identifying multi-layer coordinated perturbation on cancer pathways from TCGA data. She also talked about a recent work on integrating the 3D chromatin structures, epigenetic modification, and transcription factors to study gene regulation. Dr. Jasmine Zhou is a professor of biological sciences and computer science at the University of Southern California. Dr. Zhou is the PI of the NIH center for knowledge base on disease connections within the MAP Gen consortium. She was a recipient of several awards including an Alfred Sloan fellowship and a NSF Career award.ICIBM 2014 included four tutorial sessions that covered frontier research topics such as proteomics, metabolomics, metagenomics, single cell analysis, and next-generation sequencing and data analysis. These tutorials provided a wealth of information on these cutting-edge techniques and were well appreciated by the conference participants.\"Next-generation Sequencing and Data Analysis\" This tutorial was given by Dr. Yunlong Liu from Indiana University -Purdue University Indianapolis and Dr. Kun Huang from The Ohio State University. The tutorial introduced various aspects of RNA-seq experimental design and data analysis, including experimental consideration, transcriptome alignment, gene expression analysis, and alternative splicing analysis. Dr. Huang also demonstrated how to use popular analysis tools in the Galaxy environment.\"Proteomics and Metabolomics\" This tutorial was organized by Dr. Susan E Weintraub from The University of Texas Health Science Center at San Antonio, Dr. Steve Patrie from The University of Texas Southwestern Medical Center, and Drs. William R. Alley and Jianqiu (Michelle) Zhang from The University of Texas at San Antonio. This tutorial first provided a general introduction to mass spectrometry in proteomics research, followed by various topics that included top-down proteomics , database searching of tandem mass-spectral data for glycopeptides, and large-scale comparison of protein expression over multiple samples in LC-MS/MS. The tutorial provided a useful overview of proteome informatics for biologists, bioinformaticians, computational researchers, and alike.\"Single Cell Data Analysis\" Single cell analysis has emerged as a new paradigm for the study of biological and pathological heterogeneity. The innovative nanotechnology has made omics research possible at the single cell level, which sheds new light on the biological and pathological mechanisms and shows promise for clinical applications. This tutorial was organized by Drs. Chun-Liang Chen, Zhao Lai and Yidong Chen from The University of Texas Health Science Center at San Antonio and Dr. Chenghang Zong from Baylor College of Medicine. The tutorial introduced major elements of the single-cell analysis pipeline, as well as the technical challenges and methodology development of single cell whole genome and transcriptome sequencing.\"Metagenomics\" This tutorial was given by Dr. Chittibabu (Babu) Guda, from the University of Nebraska Medical Center and Dr. Qunfeng Dong from the University of North Texas. They provided an introduction to metagomics and demonstrated the utility of popular tools used in the analysis of metagenomic data. In addition, Dr. Guda presented MetaID, an alignment-free n-gram based method that can accurately identify microorganisms at the strain level and estimate the abundance of each microorganism in a sample. Dr. Dong also described his own research efforts on developing phylogenetic-based machine-learning method using 16S rDNA sequence data for classification, and provided step-by-step tutorials on three publicly available metagenomics software tools, Mothur, METAGENassist, and Picrust.ICIBM 2014 had six regular scientific sessions where scientists presented their recent innovative research in the areas of bioinformatics, systems biology, intelligent computing, and computational medicine. The presentations were selected through a rigorous review process handled by a program committee of more than 70 experts in the field (see Conference Organization) based on their scientific merit and technical quality. These sessions were:Session I: Gene Regulation and Protein Interaction Networks, ISession II: Gene Regulation and Protein Interaction Networks, IISession III: Algorithms in NGS Data AnalysisSession IV: Metagenomics and Bioinformatics MethodsSession V: Epigenetics and Systems BiologySession VI: Biomarker Identification MethodsThe details of each session, including session chairs, speakers, and the title and abstract of each talk are available on the conference website and in tIdentifying biomarkers to classify cancer subtypes or to predict cancer outcomes is one of the most challenging problems in cancer biology. This supplement includes impressive progress in both the development of better statistical/computational methods and also the utilization of novel molecule types as biomarkers. Ow and Kuznetsov describeA challenge in studying the genetic causes of complex diseases such as cancer and diabetes is that such diseases are usually caused by combinatorial effects of many genes, gene products, and small molecules that interact with each other to form a complex interaction network. Continuing previous years' trend ,7, severMalaria is one of the major causes of mortality around the world and the effectiveness of antimalarial drugs has been constantly challenged during the past decades due to the fast evolution of parasites that are resistant to multiple lines of drugs. Chen and Xu developeIdentification of cis-regulatory regions such as promoters, enhancers and transcription factor binding sites are fundamental tasks in deciphering the complex gene regulatory network. Several papers included in this supplement demonstrated that these elements can be more accurately predicted by integrating heterogeneous NGS data types such as DNA methylation data, histone modification marks and DNase I hypersensitivity sites, and by incorporating more accurate biophysical models of the protein-DNA interaction. For example, Hwang et al. discoverAstragalus mongolicus, one of the most important herbs used in traditional Chinese medicine, and revealed a comprehensive profile of metabolic activities among tissues for the production of bioactive compounds. The work provided valuable resources for bioengineering and in vitro synthesis of the natural compounds for medical research and for potential drug development. One major challenge in personalized medicine research is to identify the environmental factors that can alter drug response, and to investigate their molecular mechanisms. Combining bioinformatic analysis and literature mining, Philips et al. [Using RNA-seq, Chen et al. performes et al. discovers et al. , Huang eEfficient and effective computational methods are essential for solving many computational biology and computational medicine challenges. This supplement included several papers with novel computational methods, some of which are already described above in the other categories. Here we briefly describe the remaining papers in this category. In , Chen etOur sincerest thanks to the members of our Steering, Program, Publication, Workshop/Tutorial, Award, Publicity, Trainee, and Local Organization committees, as well as our numerous reviewers and volunteers, for their countless hours and energy spent to make ICIBM 2014 a success! We could not have accomplished so much without the dedication of each and every person that contributed to this conference.Sponsors National Science Foundation, University of Texas at San Antonio, University of Texas Health Science Center at San Antonio, Vanderbilt University (VU), Vanderbilt Center for Quantitative Sciences, Bioinformatics Resource Center at Vanderbilt-Ingram Cancer Center.Partners Trinity University, Texas State University at San Marcos, Indiana University -Purdue University Indianapolis, International Society of Intelligent Biological Medicine, Shanghai Center for Bioinformation Technology, China, Shanghai Institute for BioMedicine, China, and Shanghai Jiao Tong University, China.General Chairs Zhongming Zhao (Vanderbilt University) and Victor Jin .Steering Committee Yidong Chen , Ramana Davuluri (Northwestern University), Jason Ernst , Laura Elnitski , Paul Horton , Tony Hu (Drexel University), Victor Jin , Wei Li (Baylor College of Medicine), Shili Lin (Ohio State University), Zhongming Zhao (Vanderbilt University)Program Committee Chair: Yidong Chen , Co-Chair: Jeremy Edwards , Members: Kristen Anton (Dartmouth College), Yong-sheng Bai (Indiana State University), William S. Bush (Vanderbilt University), Jake Chen (Indiana University -Purdue University Indianapolis), Xue-Wen Chen (Wayne State University), Juan Cui (University of Georgia), Qinghua Cui , Youping Deng , Joshua Denny (Vanderbilt University), Jennifer M. Fettweis , Marcelo Fiszman , Jan Freudenberg , Ge Gao , Chittibabu (Babu) Guda , Yan Guo (Vanderbilt University), Steve Horvath , Tzu-Hung Hsiao , Weichun Huang, , Yang Huang (Kaiser Permanente), Yufei Huang (University of Texas at San Antonio), Jenn-Kang Hwang , Peilin Jia (Vanderbilt University), Yufang Jin (University of Texas at San Antonio), Victor Jin , Sun Kim , Judith Klein-Seetharaman , Dmitry Korkin (University of Missouri - Columbia), K.B. Kulasekera (University of Louisville), Jun Kong (Emory University), Fuhai Li , Leping Li , Liao Li (University of Delaware), Honghuang Lin (Boston University), Chunyu Liu (University of Chicago), Hongfang Liu (Georgetown University), Qi Liu (Vanderbilt University), Tianming Liu (University of Georgia), Yin Liu , Yunlong Liu (Indiana University -Purdue University Indianapolis), Zhandong Liu (Baylor College of Medicine), Xinghua Lu (University of Pittsburgh), Zhiyong Lu , Patricio A. Manque , Tabrez Mohammad , Ranadip Pal (Texas Tech University), Yonghong Peng , Horacio Perez-Sanchez , Jiang Qian (Johns Hopkins University), Thomas Rindflesch , Marylyn Ritchie (Penn State University), Bairong Shen , Alexander Statnikov , Jingchun Sun (Vanderbilt University), Wing-Kin Sung , P.S. Thiagarajan , Manabu Torii , Jun Wan (Johns Hopkins University), Edwin Wang , Jing Wang (Vanderbilt University), Junbai Wang , Yufeng Wang (University of Texas at San Antonio), Xiaoyan Wang (University of Connecticut), Chaochun Wei (Shanghai Jiao Tong University), Xiwei Wu , Yonghui Wu , Junfeng Xia , Yang Xiang (Ohio State University), Lu Xie , Hua Xu , Jianhua Xuan (Virginia Tech), Zhenqing Ye , Sungroh Yoon , Bing Zhang (Vanderbilt University), Michelle Zhang (University of Texas at San Antonio), Yanqing Zhang (Georgia State University), Min Zhao , Huiru (Jane) Zheng , W. Jim Zheng , and Dongxiao Zhu (Wayne State University).Publication Committee Chair: Jianhua Ruan (University of Texas at San Antonio), Co-Chair: Lang Li (Indiana University -Purdue University Indianapolis).Workshop/Tutorial Committee Chair: Yufang Jin (University of Texas at San Antonio), Co-Chair: Chittababu (Babu) Guda .Award Committee Chair: Yufeng Wang (University of Texas at San Antonio), Co-Chair: Hua Xu .Publicity Committee Chair: Dongxiao Zhu (Wayne State University), Co-Chair: Michelle Zhang (University of Texas at San Antonio).Trainee Committee Chair: Mario Flores (University of Texas at San Antonio), Co-Chair: Qingguo Wang (Vanderbilt University).Local Organization Committee Chair: Yufei Huang (University of Texas at San Antonio), Co-Chair: Tabrez Mohammad .The authors declare that they have no competing interests.JR, ZZ, YH, and VJ managed and participated in the peer-review of ICIBM'14 manuscripts, excluding those on which they were authors, and handled the editorial process of this supplement. YC, JE and HX supported the post-acceptance manuscript processing. JR, ZZ, YH and VJ wrote the article, which was read and approved by all authors. All authors have made substantial contribution to the collection of the manuscripts and abstracts and, thus, the success of the ICIBM 2014 conference."} +{"text": "The CDC Experience is a 1-year fellowship in applied epidemiology for third- and fourth-year medical students. Eight competitively selected fellows spend 10\u201312 months at CDC in Atlanta, Georgia, where they conduct epidemiologic analyses in areas of public health that interest them. The fellowship provides opportunities to enhance skills in research and analytic thinking, written and oral scientific presentations, and the practices of preventive medicine and public health.Through this training, fellows acquire practical knowledge for approaching population-based health problems. Graduates of The CDC Experience have an appreciation of the role of epidemiology in medicine and health and are able to apply their knowledge and skills to enhance their clinical acumen and help improve the quality of the U.S. health-care system.http://www.cdc.gov/cdcexperiencefellowship. Applications for the class of 2014\u201315 must be submitted by December 6, 2013. Questions can be e-mailed to Virginia Watson, program coordinator, at vwatson1@cdc.gov.Information on applying for The CDC Experience is available at"} +{"text": "Professor Robert Tamp\u00e9 should be added to the author byline. He should be listed as the sixth author and affiliated with Institute of Biochemistry, Biocenter, Johann Wolfgang Goethe-University, Frankfurt, Germany. The contributions of this author are as follows: Provided all synthesized trisNTA compounds.The following information is missing from the Funding section: Funding sources for Robert Tamp\u00e9 are: The German Research Foundation DFG , the BMBF (0312031/4 to R.T.) and (MODDIFSYN to R.T.) in the frame of ERA-NET NEURON supported the work."} +{"text": "Cell & Bioscience highlights review articles by Sophia Y. Tsai and Ming-Jer Tsai\u2019s research team on roles of COUP-TFII in tumor progression and metastasis and by Hui-Kuan Lin and his colleagues on posttranslational regulation of Akt in human cancer. Drs. Sophia Tsai and Ming-Jer Tsai were the 2013 Society of Chinese Bioscientists in America (SCBA) Lifetime Achievement Award winners. Dr. Hui-Kuan Lin was the 2013 SCBA Outstanding Young Investigator Award winner.This thematic issue of Cell & Bioscience, we have the privilege of publishing two review articles on cancer biology from Sophia Tsai and Ming-Jer Tsai [At the 2013 biennial international sympoium held in Xi\u2019an, China, the Society of Chinese Bioscientists in America (SCBA) recognized four of our finest members for their scientific achievements and future potential in biomedical research. The society awarded its Presidential Award to Dr. Xiao-Dong Wang , its Life-time Achievement Award to Drs. Sophia Tsai and Ming-Jer Tsai , and its Outstanding Young Investigator Award to Dr. Hui-Kuan Lin . In this issue of Drs. Sophia and Ming-Jer Tsai are prominent scientists in the field of gene regulation and gene function. Dr. Sophia Tsai holds the Gordon Cain endowed professorship in the Department of Molecular and Cellular Biology and the Department of Medicine, Baylor College of Medicine, Houston, Texas. Professor Tsai received her Bachelor and Master degrees in Chemistry from the University of Wisconsin, Madison, Wisconsin, and her PhD degree in Biochemistry from the University of California, Davis. Following her postdoctoral training at Cornell University, Ithaca and M.D. Anderson Cancer Center, Houston, she joined the Cell Biology Department at Baylor College of Medicine, Houston, TX and remained there since. Dr. Ming-Jer Tsai is Charles C. Bell Distinguished Service Professor in the Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas. Professor Tsai received his Bachelor in Science from National Taiwan University and PhD in Biochemistry from the University of California, Davis. After his postdoctoral training in the University of Texas, M.D. Anderson Cancer Center, he was recruited to the Department of Molecular and Cellular Biology, Baylor College of Medicine, as a faculty member in 1973 and remained there since. Professor Tsai is an Academician of Academia Sinica, Taiwan.Drs. Sophia and Ming-Jer Tsai have played a pivotal role in creating a new field on nuclear orphan receptors. When they and their colleagues first cloned COUP-TFI in 1989, they were among the first researchers to clone novel factors that were related to steroid receptors but not have known ligands or physiological function. Subsequently, they cloned another highly related family member, COUP-TFII. These factors became known as orphan receptors which over the years have expanded the steroid receptor family to a superfamily of 48 members in humans. They showed that COUP-TFI and II are essential for cell fate determination during embryonic development and established them as early critical players in normal development, controlling cell-fate specification, neural development, organogenesis, angiogenesis and metabolism. Dr. Tsai\u2019s team also showed that dysregulation of COUP-TFII is the major underlying reason for serious diseases, such as congenital diaphragmatic hernia, congenital heart defects, tumorigenesis and heart failure. Their review article is focusing on the roles of COUP-TFII in tumorigenesis and tumor metastasis.Dr. Hui-Kuan Lin is currently holding an Associate Professor position and R. Lee Clark scholar in the Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas. Dr. Lin received his Bachelor\u2019s and Master\u2019s degrees in Pharmacy and Pharmacology from National Taiwan University, Taipei, Taiwan 1993 and 1995, respectively. He then moved to the University of Rochester, Rochester, New York for his Ph.D study of Cancer Biology until his graduation in 2002. Afterwards, Dr. Lin went to the Memorial Sloan-Kettering Cancer Center, New York, for his post doctoral training in Cancer Biology and Genetics with Dr. Pier Paolo Pandolfi. He was recruited as a faculty member by the University of Texas Health Science Center and MD Anderson Cancer Center, Houston, Texas in 2007 and promoted to Associate Professor in 2011.Dr. Lin is an outstanding junior investigator with his research focusing on the roles of posttranslational modifications of proteins in cancer signaling. Dr. Lin made important contributions to the understanding of key signal pathways, such as PI3K/PTEN/Akt and TGF-beta, involved in regulation of cancer development. His recent work has been directed to understand the role of posttranslational regulation of protein factors in cancer development. In the review article, Dr. Lin described Akt activation through posttranslational modifications, including phosphorylation, OGlcNAcylation, ubiquitination, SUMOylation and acetylation.SCBA appreciates scientific contributions by its outstanding members like Drs. Sophia and Ming-Jer Tsai, and Hui-Kuan Lin since these contributions will eventually benefit all human beings for better health and life."} +{"text": "There is an error in the Correction published on March 31, 2016. The ninth author\u2019s name is: Eun-Taek Han. The correct text is:There is an error in the affiliation for the ninth author. Eun-Taek Han is not affiliated with #3 but with #2 Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do, Republic of Korea.The publisher apologizes for the error."} +{"text": "Cancer is a complex whole-body disease that alters in the levels of gene, protein, and metabolite, and that involves multi-factors, multi-processes and multi-consequences. Individual variation is involved in each stage of prediction/prevention, early-stage diagnosis/therapy, and late-stage diagnosis/therapy. The development of omics and systems biology has promoted one to gradually change paradigms in oncology from traditional single factor strategy to multi-parameter systematic strategy. The therapeutic model of cancer has changed from the general radiotherapy and chemotherapy to personalized strategy. The development of PPPM will substantially change the understanding, prediction, prevention, and therapeutic model of cancer from a systematical and comprehensive point of view in the future."} +{"text": "Childhood is important and critical period in human life. The foundation of ego is shaped in childhood. Play therapy is one of the successful strategies to help children with inner conflicts problems. This method of psychotherapy is base on the normal learning processes of children, provides solutions to relieve feelings of stress, and expands self-expression. Group play therapy can enhance the self-awareness, self- regulation, social communication, empathy and adoptability in children.Present study investigated the effects of play therapy on relational and emotional skills of pre-school children. For this purpose, the total numbers of 372 pre-school children were randomly selected, and divided into two equal groups (case and control). In next step, the BUSSE-SR methodology was used for evaluation and comparison of self-awareness, self-regulation, social interaction, empathy, adoptability, and control groups. Pre-test were performed for both groups and case group was involved in-group play therapy. According to the results of post-test, correlation of variables between case-control groups was examined by multivariate analysis of covariance.Frequency of boys and girls in our sample were 51.3 and 48.7 percent, respectively. The mean age of children was 5.1\u00b10.6 year. According to the results of present study, play therapy significantly enhanced the social-emotional skills (P< 0.001). Our findings are consistent with the results of previous studies in other nations with different environmental and cultural properties. In conclusion, it seems that play therapy can be used in pre-school centers to help children learn problem-solving skills and communicate with others. Childhood is the important and critical period of life with critical effects on personality of individuals. In other hand, it is best period to help children learn adaptive behaviors and effective communicational skills . In nextSocial-emotional development includes a set of skills such as self-awareness, understanding of others emotions, emotional management, emotional expressions in a constructive manner, self-regulation, stabled communications . Social-Play therapy helps children with problems to provide a safe distance from their psychological problems and express appropriate interactions . Play thIn present study, we examined the impact of group play therapy on self-awareness, self-regulation, social interaction, empathy, and adoptability of pre-school children in Tehran, Iran.In present study, we used pre and post-test methodology for comparison the differences of self-awareness, self-regulation, social interaction, empathy, and adoptability between case and control groups. According to the number of pre-school children in the area understudy in Tehran (11697) and based on Morgan table, a random distributed population including 372 individuals were selected. We divided sample population randomly in two equal groups including case and control (186 in each group). In next step, BUSSE-SR social-emotional questionnaire was used . The queIn first step, we examined both groups by pretest. For this purpose, parents of selected children filled the questionnaire according to the instructions to fill. We collected questionnaires after 2 days and analyzed. The case group was involved in directed social-emotional play therapy during the fifteen sessions of 90 min (Three sessions a week). In directed play therapy, the therapist was responsible for selection and leadership of the games. Children were encouraged to express their problems and find solutions. Then, the post-test was carried out for both groups by written. We performed analysis of covariance (ANCOVA) by SPSS 17 to compare mean results and control groups.According to the findings of this study, 51.3and 48.7 percent of children were boys and girls respectively. The mean of age was 5.1years old. The mean of height was 108.3 centimeters for girls and 110.1 centimeters for boys. In addition, the mean of weight was 17.3 kg for girls and 18.9 kg for boys. All the children were mentally and physically normal. We did not find any significant differences in mean scores of two groups in pre-test .As we can see in In present study, we considered the impact of directed group play therapy on improvement of social-emotional skills of pre-school children. Our hypothesis was the play therapy method could be an effective and general approach in training of children to establish communications, express thoughts and feelings, and solve their problems . We studThe efficacy of group play therapy has been considered in some studies, and they indicated that this method is effective to prevent or resolve psychosocial problems . For insConsidering the results of present study and other research results, it can be concluded that play therapy provides environment where children can measure their own abilities, express themselves, and learn how to use their knowledge to take maximum advantages of their capacities. Finally, to extend the application of group game therapy method benefits and enhance children\u2019s social-emotional skills training of related skills to teachers in childhood education centers are recommended."} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This study was partially funded by the EU-FP7 MEPHITIS project, by FEDER funds through COMPETE (FCOMP-01-0124-FEDER-014284) and by National Funds through FCT\u2014Foundation for Science and Technology in the scope of the research projects PTDC/BIA-BCM/72251/2006, PTDC/BIA-BCM/64745/2006 and PTDC/BIA-GEN/110383/2009. The Institute of Electronics and Telematics Engineering of Aveiro (IEETA) supported the development of the Anaconda software package. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The Funding statement for this article is incorrect. The correct Funding statement is provided below:Funding: This work was supported by grant W911NF-13-1-0199 from the US Army Research Office. KHT is a Cystic Fibrosis Foundation Postdoctoral Research Fellow. MW is a Burroughs Wellcome Investigator in the Pathogenesis of Infectious Disease. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "It has been well-established, both by population genetics theory and direct observation in many organisms, that increased genetic diversity provides a survival advantage. However, given the limitations of both sample size and genome-wide metrics, this hypothesis has not been comprehensively tested in human populations. Moreover, the presence of numerous segregating small effect alleles that influence traits that directly impact health directly raises the question as to whether global measures of genomic variation are themselves associated with human health and disease.We performed a meta-analysis of 17 cohorts followed prospectively, with a combined sample size of 46,716 individuals, including a total of 15,234 deaths. We find a significant association between increased heterozygosity and survival (P\u2009=\u20090.03). We estimate that within a single population, every standard deviation of heterozygosity an individual has over the mean decreases that person\u2019s risk of death by 1.57%.This effect was consistent between European and African ancestry cohorts, men and women, and major causes of death (cancer and cardiovascular disease), demonstrating the broad positive impact of genomic diversity on human survival.The online version of this article (doi:10.1186/s12863-014-0159-7) contains supplementary material, which is available to authorized users. With the advent of genome-wide association studies (GWAS), and more recently whole-exome and whole-genome sequencing, remarkable progress has been made in elucidating the genetics of complex traits, with numerous genetic variants each explaining a small fraction of the variance ,2. The pVarious heterozygosity metrics have been proposed . The hetrequency ,14, up-wTo test for the effect of genome-wide heterozygosity on survival, we performed a meta-analysis of 17 cohorts followed prospectively, with a combined sample size of 46,716 individuals, including a total of 15,234 deaths included covariates for Body Mass Index (BMI) at first visit and first ten principal components, and the 'strata' function for sex, education level , income level (defined by cohorts), and center of DNA collection within cohorts. The CoxPH model was set up so that the outcome was age at study entry, age at study exit, and a binary variable coding state of death . Age is measured in units of years, but is accurate to the nearest day.For the meta-analysis, significance was determined by Stouffer's method calculatInstitutional Review Board approvals were obtained by each participating ARIC study center and the coordinating center (University of NC), and the research was conducted in accordance with the principles described in the Helsinki Declaration. All subjects in the ARIC study gave informed consent. For more information see dbGaP Study Accession: phs000280.v2.p1. JHSPH IRB number H.34.99.07.02.A1. Manuscript proposal number MS1964.HealthABC Human subjects protocol UCSF IRB is H5254-12688-11.CHS was approved by institutional review committees at each site, the subjects gave informed consent, and those included in the present analysis consented to the use of their genetic information for the study of cardiovascular disease. It is the position of the UW IRB that these studies of de-identified data, with no patient contact, do not constitute human subjects research. Therefore we have neither an approval number, nor an exemption.IRB permission to conduct genetics-related work in the Health and Retirement Study (HRS) is granted under the project title, \"Expanding a National Resource for Genetic Research in Behavioral & Health Science\" (HUM00063444). The IRB that approved this project is the Health Sciences and Behavioral Sciences Institutional Review Board at the University of Michigan. No manuscript proposal is required for use of HRS data.Inchianti ethics review statement: The study protocol was approved by the Italian National Institute of Research and Care of Aging Institutional Review and Medstar Research Institute .The Religious Orders Study (ORA# 91020181) and the Rush Memory and Aging Project (ORA# 86121802) were approved by the Institutional Review Board of Rush University Medical Center. Written informed consent was obtained from all the participants.The SHIP study followed the recommendations of the Declaration of Helsinki. The study protocol of SHIP was approved by the medical ethics committee of the University of Greifswald. Written informed consent was obtained from each of the study participants. The SHIP study is described in PMID: 20167617.The Rotterdam Study has been approved by the medical ethics committee according to the Population Study Act Rotterdam Study, executed by the Ministry of Health, Welfare and Sports of the Netherlands. A written informed consent was obtained from all participants.The Boston University Medical Campus Institutional Review Board approved the FHS genome-wide genotyping (protocol number H-226671) and genetic investigation of aging and longevity phenotypes (protocol number H-24912).The Age, Gene/Environment Susceptibility Reykjavik Study has been funded by NIH contract N01-AG-12100, the NIA Intramural Research Program, Hjartavernd (the Icelandic Heart Association), and the Althingi (the Icelandic Parliament). The study is approved by the Icelandic National Bioethics Committee, (VSN: 00\u2013063) and the Data Protection Authority. The researchers are indebted to the participants for their willingness to participate in the study.Ethics permission for the LBC studies was obtained from the Multi-Centre Research Ethics Committee for Scotland (MREC/01/0/56) and from Lothian Research Ethics Committee (LBC1936: LREC/2003/2/29 and LBC1921: LREC/1998/4/183). The research was carried out in compliance with the Helsinki Declaration. All subjects gave written, informed consent."} +{"text": "Identifying, developing, and testing implementation strategies are important goals of implementation science. However, these efforts have been complicated by the use of inconsistent language and inadequate descriptions of implementation strategies in the literature. The Expert Recommendations for Implementing Change (ERIC) study aimed to refine a published compilation of implementation strategy terms and definitions by systematically gathering input from a wide range of stakeholders with expertise in implementation science and clinical practice.Purposive sampling was used to recruit a panel of experts in implementation and clinical practice who engaged in three rounds of a modified Delphi process to generate consensus on implementation strategies and definitions. The first and second rounds involved Web-based surveys soliciting comments on implementation strategy terms and definitions. After each round, iterative refinements were made based upon participant feedback. The third round involved a live polling and consensus process via a Web-based platform and conference call.Participants identified substantial concerns with 31% of the terms and/or definitions and suggested five additional strategies. Seventy-five percent of definitions from the originally published compilation of strategies were retained after voting. Ultimately, the expert panel reached consensus on a final compilation of 73 implementation strategies.This research advances the field by improving the conceptual clarity, relevance, and comprehensiveness of implementation strategies that can be used in isolation or combination in implementation research and practice. Future phases of ERIC will focus on developing conceptually distinct categories of strategies as well as ratings for each strategy\u2019s importance and feasibility. Next, the expert panel will recommend multifaceted strategies for hypothetical yet real-world scenarios that vary by sites\u2019 endorsement of evidence-based programs and practices and the strength of contextual supports that surround the effort.The online version of this article (doi:10.1186/s13012-015-0209-1) contains supplementary material, which is available to authorized users. Thi clarity ,10. Idioer time) . Implemeer time) -7,12. Seer time) ,14. Takee.g., [et al. [et al. [e.g. [e.g. [e.g. [e.g. [e.g. [e.g. [are purposely narrow in scope, focusing on strategies with known evidence on effectiveness, e.g. -22; specg. [e.g. -25; strag. [e.g. ,27; \u201cexeg. [e.g. ,29; one g. [e.g. ; or one g. [e.g. ,31. The A number of taxonomies of implementation strategies have been developed, in part, to address these shortcomings pertaining to the published literature, e.g., ,15-18. P [et al. reviewed [et al. note thaet al. [et al. [et al.\u2019s [e.g., [In response to those limitations, Powell et al. proposedet al. . This raet al. builds u [et al. review b [et al. . We purset al.\u2019s compilats [e.g., ,10,32, ts [e.g., .Implementation Science, implementation research coordinators for the VA Quality Enhancement Research Initiatives (QUERIs) [i.e., Eastern through Pacific) in order to minimize scheduling conflicts for the live Webinar (described below). Ultimately, we recruited a panel of 71 experts (see \u201cWe employed a purposive sampling procedure that beg(QUERIs) , and fac(QUERIs) . NomineeThe modified Delphi process had threet al.\u2019s [et al.\u2019s [Fifty-seven experts completed the Round 1 Web-based survey. Section one of the Round 1 survey listed terms and definitions from Powell et al.\u2019s publisheet al.\u2019s compilate.g., 72% of panelists made no comment). The core definitions from the original compilation [Forty-three experts completed the Round 2 Web-based survey, which included the implementation strategy terms and definitions from Round 1 along with a summary of the panelists\u2019 comments and suggestions regarding additional strategies. This included both a qualitative summary and, where possible, a quantitative characterization of participants\u2019 Round 1 responses was not obtained in the first runoff vote, then the top two alternatives from the first runoff round would advance to a final runoff round to determine the winner. If a tie between the original and alternative definition occurred in the runoff round, the definition already published in the literature was retained. These same voting procedures were applied to the additional strategies proposed by the expert panel in Rounds 1 and 2 of the Delphi process; however, the approval poll also included an option for the proposed strategy to be rejected if a supermajority (\u226560%) of panelists deemed the strategy unworthy of inclusion. Figure\u00a0Following open discussion, the second stage of voting involved \u201crunoff voting\u201d, in which participants selected only their top choice. If only two alternatives were presented, the definition receiving the most votes was declared the winner. If three or more alternatives were presented and a majority yielded a clear winner; however, in two cases, no strategy received over 60% of the vote in the approval voting stage and in another case there was a tie between two strategies, each receiving 66% of the votes. In these situations, the participants discussed their thoughts and concerns, after which the runoff vote successfully identified a winning definition.The majority of the terms and definitions (69%) from the Powell et al. compilatFor the 21 alternative definitions suggested, an alternative definition was selected 81% of the time and the original definition was maintained 19% of the time. One of the advantages of approval voting was determining the acceptability of the original definitions even when alternatives were thought to be superior. In each of the 17 times in which an alternative was ultimately selected, the original definitions failed to reach the supermajority approval level of 60% .Each of the five new strategies that the panel proposed was maintained in some form. Panelists had the opportunity to reject the proposed additions, but on average, across the five strategies, 84% of panelists voted to retain the new strategy (range 100 to 71%). Each of the new strategies had an initial proposed definition in Round 1. Panelists had the opportunity to suggest alternative definitions in Round 2. In two cases , no alternative definitions were proposed, and the new definition was retained with approval votes of 71 and 73%, respectively. In one case (\u201cidentify early adopters\u201d) the alternative definition won in the approval vote. Finally, in two cases (\u201cfacilitation\u201d and \u201cpromote adaptability\u201d), the original new definition was selected over the alternatives in the runoff vote.et al. [The final compilation included 73 discrete strategies in hypothetical VA mental health clinic settings that vary on certain contextual characteristics [The subsequent stages of the ERIC project will fureristics . This stet al. [As Powell et al. cautioneWe note that while our attempt was to identify discrete strategies involving one action or process, the included strategies vary in their level of complexity. In fact, active research agendas have focused on determining the essential components of many of these \u201cdiscrete\u201d implementation strategies, such as audit and feedback , learninet al. [The ERIC compilation consolidated discrete implementation strategies that have been identified through other taxonomies and reviews , but we acknowledge that broader international participation would have been ideal. This may have implications for the content of the compilation, as we discuss below. Third, it is possible that in-person meetings may have generated more nuanced discussions of strategy terms and definitions; however, the asynchronous, online process had the advantage of allowing a wide range of implementation and clinical experts to participate and also ensured anonymity of responses, which limited the possibility of participants simply yielding to the majority opinion in Rounds 1 and 2. Finally, as noted in the \u201ce.g., technical difficulties, other distractions, not finding any of the strategy terms and definitions appropriate), we cannot be certain as to why participants abstained or about whether or not this could have impacted the final results in cases in which voting results were extremely close.There are several limitations related to the process of generating this compilation. First, had we used a different taxonomy of implementation strategies as a starting point, the modified Delphi process may have yielded different results. However, the original Powell et al. compilat. [e.g., -17, incre.g., the Consolidated Framework for Implementation Research [i.e., the defining characteristics of the implementation strategies); 2) causal mechanisms ; 3) mode of delivery or practical application ; and 4) intended target . Lastly, while we are not aware of evidence that would suggest that the strategies in this compilation would not be applicable to many different contexts, it is possible that some of the strategies may be more applicable to US or North American settings given the focus of the ERIC project and the composition of the expert panel. Engaging a broader international panel may have revealed additional strategies that are applicable to health-care systems that are organized differently or to settings that are not similarly resourced. The fact that the original compilation drew from taxonomies developed in contexts other than the US, e.g., [There are also limitations related to the content of the refined compilation. First, the evidence base for each strategy was not considered because the purpose of this work was to identify the range of potential options available. Second, the strategies were not explicitly tied to relevant theories or conceptual models. The compilation\u2019s utility would be enhanced by linking each strategy to the domains of prominent conceptual frameworks . FurtheResearch to betteS, e.g., ,17 may het al.\u2019s [This research advances the field by improving the conceptual clarity, relevance, and comprehensiveness of discrete implementation strategies that can be used in isolation or combination in implementation research and practice. The utility of this compilation will be extended in subsequent stages of the ERIC study. We conclude by echoing Powell et al.\u2019s caution We would like to acknowledge the contributions of each member of the expert panel: Greg Aarons, University of California, San Diego; Mark Bauer, Harvard University and US Department of Veterans Affairs; Rinad Beidas, University of Pennsylvania; Sharon Benjamin, Alchemy; Ian Bennett, University of Pennsylvania; Nancy Bernardy, Dartmouth College and US Department of Veterans Affairs; Amy Bohnert, University of Michigan and US Department of Veterans Affairs; Melissa Brouwer, McMaster University; Leo Cabassa, Columbia University; Martin Charns, Boston University and US Department of Veterans Affairs; Amy Cohen, US Department of Veterans Affairs; Laurel Copeland, Scott and White Healthcare and US Department of Veterans Affairs; Torrey Creed, University of Pennsylvania; Jill Crowley, US Department of Veterans Affairs; Geoff Curran, University of Arkansas for Medical Sciences and US Department of Veterans Affairs; Laura Damschroder, University of Michigan and US Department of Veterans Affairs; Teresa Damush, Indiana University and US Department of Veterans Affairs; Afsoon Eftekhari, US Department of Veterans Affairs; Rani Elwy, Boston University and US Department of Veterans Affairs; Bradford Felker, University of Washington and US Department of Veterans Affairs; Erin Finley, University of Texas Health Science Center San Antonio and US Department of Veterans Affairs; Hildi Hagedorn, University of Minnesota and US Department of Veterans Affairs; Alison Hamilton, University of California, Los Angeles and US Department of Veterans Affairs; Susanne Hempel, RAND; Timothy Hogan, University of Massachusetts and US Department of Veterans Affairs; Bradley Karlin, Education Development Center and US Department of Veterans Affairs; Ira Katz, US Department of Veterans Affairs; Jacob Kean, Indiana University and US Department of Veterans Affairs; Shannon Kehle-Forbes, University of Minnesota and US Department of Veterans Affairs; Amy Kilbourne, University of Michigan and US Department of Veterans Affairs; Kelly Koerner, Evidence-Based Practice Institute; Sarah Krein, University of Michigan and US Department of Veterans Affairs; Julie Kreyenbuhl, University of Maryland and US Department of Veterans Affairs; Kurt Kroenke, Indiana University and US Department of Veterans Affairs; Marina Kukla, Indiana University-Purdue University Indianapolis and US Department of Veterans Affairs; Sara Landes, University of Washington and US Department of Veterans Affairs; Martin Lee, University of California, Los Angeles and Prolacta Bioscience; Cara Lewis, Indiana University-Bloomington; Julie Lowery, University of Michigan and US Department of Veterans Affairs; Brian Lund, US Department of Veterans Affairs; Aaron Lyon, University of Washington; Natalie Maples, University of Texas Health Science Center San Antonio; Stephen Marder, University of California, Los Angeles and US Department of Veterans Affairs; Monica Matthieu, Saint Louis University and US Department of Veterans Affairs; Geraldine McGlynn, US Department of Veterans Affairs; Alan McGuire, Indiana University-Purdue University Indianapolis and US Department of Veterans Affairs; Allison Metz, University of North Carolina; Amanda Midboe, US Department of Veterans Affairs; Edward Miech, Indiana University and US Department of Veterans Affairs; Brian Mittman, US Department of Veterans Affairs; Laura Murray, Johns Hopkins University; Princess Osei-Bonsu, US Department of Veterans Affairs; Richard Owen, University of Arkansas for Medical Sciences and US Department of Veterans Affairs; Louise Parker, University of Massachusetts Boston; Mona Ritchie, US Department of Veterans Affairs; Craig Rosen, Stanford University and US Department of Veterans Affairs; Anju Sahay, US Department of Veterans Affairs; Susanne Salem-Schatz, Health Care Quality Initiatives; Anne Sales, University of Michigan and US Department of Veterans Affairs; Mark Snowden, University of Washington; Leif Solberg, Health Partners; Sharon Straus, University of Toronto; Scott Stroup, Columbia University; Jane Taylor, CHAMP; Carol VanDeusen Lukas, Boston University and US Department of Veterans Affairs; Dawn Velligan, University of Texas Health Science Center San Antonio; Robyn Walser, University of California, Berkeley and US Department of Veterans Affairs; Shannon Wiltsey-Stirman, Boston University and US Department of Veterans Affairs; Gordon Wood, US Department of Veterans Affairs; Kara Zivin, University of Michigan and US Department of Veterans Affairs; and Cynthia Zubritsky, University of Pennsylvania.aAs Wensing et al. [e.g., implementation science, knowledge translation research, improvement science, research utilization, delivery science, quality improvement, etc.). While each of these traditions \u201cbring their own nuances to the area\u2026the reality is that there are far more commonalities in the research conducted under these different names than differences\u201d [e.g., knowledge translation strategies or interventions, quality improvement strategies, implementation interventions, strategies to increase research utilization, etc.), we believe that the compilation described in this paper is likely to be applicable to the research and practice occurring under these different names. Indeed, the original Powell et al. [g et al. note, therences\u201d . Thus, wl et al. compilatl et al. and \u201cknol et al. among ot"} +{"text": "Porphyromonas gingivalis in the Lung. The correct citation is: Benedyk M, Byrne DP, Glowczyk I, Potempa J, Olczak M, Olczak T, et al. (2015) Pyocyanin, a Contributory Factor in Haem Acquisition and Virulence Enhancement of Porphyromonas gingivalis in the Lung. PLoS ONE 10(2): e0118319. doi:10.1371/journal.pone.0118319The word \u201cPyocyanin\u201d is misspelled in the article title. The correct title is: Pyocyanin, a Contributory Factor in Haem Acquisition and Virulence Enhancement of The following information is missing from the Funding section: The Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University is a partner of the Leading National Research Center (KNOW) supported by the Ministry of Science and Higher Education."} +{"text": "Infectious diseases of poverty (IDoP) disproportionately affect the poorest population in the world and contribute to a cycle of poverty as a result of decreased productivity ensuing from long-term illness, disability, and social stigma. In 2010, the global deaths from HIV/AIDS have increased to 1.5 million and malaria mortality rose to 1.17 million. Mortality from neglected tropical diseases rose to 152,000, while tuberculosis killed 1.2 million people that same year. Substantial regional variations exist in the distribution of these diseases as they are primarily concentrated in rural areas of Sub-Saharan Africa, Asia, and Latin America, with geographic overlap and high levels of co-infection. Evidence-based interventions exist to prevent and control these diseases, however, the coverage still remains low with an emerging challenge of antimicrobial resistance. Therefore, community-based delivery platforms are increasingly being advocated to ensure sustainability and combat co-infections.Because of the high morbidity and mortality burden of these diseases, especially in resource-poor settings, it is imperative to conduct a systematic review to identify strategies to prevent and control these diseases. Therefore, we attempted to evaluate the effectiveness of one of these strategies, that is community-based delivery for the prevention and treatment of IDoP. In this paper, we describe the burden, epidemiology, and potential interventions for IDoP. In subsequent papers of this series, we describe the analytical framework and the methodology used to guide the systematic reviews, and report the findings and interpretations of our analyses of the impact of community-based strategies on individual IDoPs. Please see Additional file The Global Burden of Disease Study 2010 reports an increase of 111,000 deaths globally attributable to malaria and neglected tropical diseases (NTDs) in the last two decades, with substantial regional variations and Sub-Saharan Africa accountable for most of the premature mortalities [talities -6. NTDs talities -11. It italities .IDoP disproportionately affect the poorest populations in the world and contribute to a cycle of poverty as a result of decreased productivity ensuing from long-term illness, disability, and social stigma ,5. The aA large proportion of these infectious diseases in low- middle- income countries (LMICs) are entirely avoidable or treatable with existing medicines or interventions . EffectiNTD are a group of 17 bacterial, parasitic protozoal, and viral infections that are chronic and particularly endemic amongst the population in tropical and subtropical regions promotes the use of five public health strategies to control, eliminate, and eradicate NTDs. These include preventive chemotherapy; innovative and intensified disease-management; vector control and pesticide management; provision of safe drinking water, basic sanitation and hygiene, and education; and veterinary public health services . Mass drThe WHO recommends periodic preventive treatment with anthelmintics for all at-risk people living in endemic areas to reduce morbidity by reducing the worm burden. Large-scale and successful control activities implemented during 2001\u20132010 demonstrate the feasibility of mass deworming, and these experiences have informed the development of tools to facilitate its implementation . Of the Key interventions recommended by the WHO to prevent and control malaria include prompt and effective treatment with artemisinin-based combination therapies; the use of insecticide-treated nets (ITNs); and indoor residual spraying (IRS) with insecticide to control vector mosquitoes. In the past decade, the percentage of households owning at least one ITN in Sub-Saharan Africa reached an estimated 53% by 2011 and remained at 53% in 2012. It must be noted however that this is greatly challenged by the limited deliveries of ITNs and increasing mosquito resistance to insecticides . In 2011Strategies for HIV prevention involve risk-reduction through education and counseling. The WHO has recommended key approaches which include condom use, testing and counseling, male circumcision, preventive antiretroviral therapy (ART), harm reduction for injecting drug users, and elimination of mother-to-child transmission (MTCT) of HIV ,37. In 2TB is preventable as well as curable, and its transmission could be prevented by prompt identification and treatment of the infection. The WHO is working to dramatically reduce the burden of TB and halve TB deaths and prevalence by 2015 through its Stop TB Strategy and by supporting the Global Plan to Stop TB. Between 1995 and 2011, 51 million people were successfully treated for TB in countries that had adopted the WHO strategy, saving 20 million lives . There hA large proportion of infectious diseases in LMICs are entirely avoidable or treatable with existing medicines or interventions which are also highly cost effective, however, their delivery to the affected populations has proven very difficult due to weak health systems and infrastructures . AnotherEffective delivery of proven interventions require a variety of components ranging from training health workers, effective use of epidemiological data, proper delivery of safe medicines and commodities, accurate monitoring and evaluation, and providing feedback to the community. Successful implementation requires a positive interrelation between programs for disease control and the health system at large. Global health initiatives have created a complex health system with an increasing number of actors entering the field and implementing diverse health systems strategies . These hCommunity-based interventions (CBIs) have the potential to overcome the barriers of access and availability and, if adequately equipped and supported by parallel structures, can make a significant impact on reducing the burden of IDoP -44. HoweIn order to evaluate the effectiveness of CBIs, we developed an analytical framework and conducted systematic reviews of the existing studies focusing on CBIs for prevention and control of helminthic and non-helminthic NTDs, malaria, TB, and HIV/AIDS compared to the routine healthcare delivery. For this review, we categorized NTDs into helminthic and non-helminthic diseases, and reported the findings accordingly in separate papers. Helminthic diseases included soil-transmitted helminthiasis along with schistosomiasis, lymphatic filariasis, onchocerciasis, and dracunculiasis. Non-helminthic diseases included dengue, African trypanosomiasis, chagas, leishmaniasis, trachoma, leprosy, and buruli ulcer. In this series of eight papers, we describe the analytical framework and methodology used for the systematic reviews, and report findings on the effectiveness of CBIs for the prevention and control of helminthic NTDs, non-helminthic NTDs, malaria, HIV/AIDS, and tuberculosis. In the final paper, we propose a way forward.ART: Antiretroviral therapy; CBI: Community-based intervention; CHW: Community health worker; DALY: Disability-adjusted life year; HIV/AIDS: Human immunodeficiency virus/acquired immunodeficiency syndrome; IDoP: Infectious disease of poverty; IPTp: Intermittent preventive therapy during pregnancy; IRS: Indoor residual spraying; ITN: Insecticide-treated net; LF: Lymphatic filariasis; LMIC: Low- middle- income country; MDA: Mass drug administration; MDG: Millennium development goal; MDR-TB: Multi drug resistant tuberculosis; MTCT: Mother-to-child transmission; NTD: Neglected tropical disease; RBM: Roll back malaria; STH: Soil-transmitted helminthiasis; TB: Tuberculosis; UN: United nations; WHA: World health assembly; WHO: World health organization.The authors declare that they have no financial or non-financial competing interests.ZAB and JS were responsible for designing and coordinating the review. All authors contributed, read and approved the final manuscript.Multilingual abstracts in the six official working languages of the United Nations.Click here for file"} +{"text": "Computers are incredibly fast, accurate, and stupid.Human beings are incredibly slow, inaccurate, and brilliant.Together they are powerful beyond imagination(Einstein never said that ).The life sciences, biomedicine and health care are increasingly turning into a data intensive science -4. PartiDue to the increasing trend towards personalized and precision medicine , biomedty\" data and the 1EB=1\u00d710Byte. Lasfourth paradigm in the investigation of nature - after empiricism, theory and computation . M. M21]. Minteraction, due to the fact that it is the human end user who possesses the problem solving intelligence , m, minteralligence . At the lligence , 31], in in34] in\"A wealth of information creates a poverty of attention and a need to allocate that attention efficiently among the overabundance of information sources that might consume it\" , , .Figure Many different biological species deliver large amounts of data, together with the enormous complexity of medicine per se and the \u2022 Heterogeneous data sources (need for data fusion);\u2022 Complexity of the data ;\u2022 Noisy, uncertain data, dirty data, the discrepancy between data-information-knowledge (various definitions), Big data sets .In comparison to research systems, commercially available information systems have only limited data fusion capabilities, if any at all . It is aThe problem of merging multiple data sets concerning common entities is frequently encountered in KDD, often called the Merge/Purge problem, it is difficult to solve both in scale and accuracy . CleansiMany data mining methods are designed for collections of objects well-represented in rigid tabular formats. However, besides massive sets of unstructured information and non-standardized information (text) -76, we aAdvanced data mining approaches include:1) graph-based data mining , 79], , , 81],,81],2) entropy-based data mining ,82, 83--83-85, a3) topological data mining ,87.We emphasize that these approaches are interdisciplinary and complementary albeit having common goals, and have been proven useful to perform translational research, e.g., ,82,84,85In particular, entropy-based graph analysis is based on using information theory and graph theory. Generally, information theory relates Finally, the results gained by the application of sophisticated algorithms in high dimensional spaces in area 3 must be mapped back to as the mapping from the higher into the lower dimensional space, a process that always suffers the danger of modelling artefacts. Although Visualization is a mature field with a background of several decades, there are still a lot of challenging and open research issues, especially in the context of interactive data mining with application to the biomedical domain. A major issue is the absence of a complete toolset that supports all analysis tasks within a biomedical workflow, including the many steps of data preprocessing [We can say that, while our world is highly dimensional mathematically, we can only perceive lower dimensions. This leads to the definition of visualization ocessing . It is vocessing -102, - tocessing ,104; Thiocessing -107.Whenever we deal with biomedical data issues of privacy, data protection, data security and data safety and the fair use of data are of paramount importance , includiSome additional aspects to consider include:Building a project consortium comprising of experts with complementary expertise but common interests is a success factor in each project. Bringing together domain experts from diverse areas in a cross-disciplinary manner is a challenge to stimulate fresh ideas and encouraging multi-disciplinary work . For exathe measures in data mining [As we broaden workflows for data mining, we have to expand metrics used to evaluate our results. It is no longer sufficient to focus on performance metrics, such as ROC , accuraca mining ), one mua mining , for exaa mining , interpra mining , consequhttp://www.worldcommunitygrid.org), [As our computing machinery evolves, from large main-frame servers to multi-core CPU/GPU clusters we need to optimize data mining algorithms, processes and workflows to best fit the environment. The potential of so-called On-Demand Hardware along with the Software as a Service (SAAS) paradigm can no lid.org), .To measure the quality of data mining approaches, the production of benchmarks it very important. These data sets can be used as so-called gold-standards and Knowledge Discovery from Data (KDD). A proverb attributed perhaps incorrectly to Albert Einstein illustrates this perfectly: \"Computers are incredibly fast, accurate, but stupid. Humans are incredibly slow, inaccurate, but brilliant. Together they may be powerful beyond imagination\".Andreas Holzinger is head of the Research Unit Human-Computer Interaction, Institute for Medical Informatics at the Medical University Graz, Lead at the HCI-KDD network, head of the first Austrian IBM Watson Think Group, Associate Professor of Applied Informatics at the Faculty of Computer Science, Institute of Information Systems and Computer Media and Lecturer at the Institute of Genomics and Bioinformatics at Graz University of Technology. He serves as consultant for the Canadian, Swiss, French and Dutch Government, for the German Excellence Initiative and as national expert in the European Commission (Lisbon Delegate 2000). Andreas, born 1963, started as an apprentice in IT in 1978; while working as an industrial engineer, he resumed a parallel second-chance education, finished his PhD in Cognitive Science in 1997 and completed his second doctorate (Habilitation) in Computer Science in 2003. Since 1999 participation in leading positions in 30+ R&D multi-national projects, budget 3+ MEUR; 300+ publications, >4000+ citations. Andreas was Visiting Professor in Berlin, Innsbruck, Vienna, London, and Aachen. He is passionate on bringing together Human-Computer Interaction (HCI) and Knowledge Discovery/Data Mining (KDD), with the goal of supporting human intelligence with machine intelligence - to discover new, previously unknown insights into complex biomedical data. http://www.hci4all.atMatthias Dehmer is currently head of the Division for Bioinformatics and Translational Research at UMIT, Austria and a professor of discrete mathematics. He studied mathematics and computer science at the University of Siegen (Germany) where he graduated in 1998. Between 1998 and 2002, he held positions as a mathematical researcher and a business consultant in industry. He joined in 2002 the Department of Computer Science at Darmstadt University of Technology and obtained a PhD in computer science. From 2005 to 2008, he held several research positions at the University of Rostock (Germany), Vienna Bio Center (Austria), Vienna Technical University (Austria) and University of Coimbra . Finally, he obtained his habilitation in applied discrete mathematics from the Vienna University of Technology. He has been the head of the Division for Bioinformatics and Translational Research at UMIT, Austria. He has published over 160 publications in applied mathematics and computer science. Moreover, he is an editor of the book series \"Quantitative and Network Biology\", Wiley-VCH. He organized and co-organized several international scientific conferences and workshops in USA. Also, he recently got a member of the editorial board of Scientific Reports (Nature) and PLoS ONE. http://www.dehmer.orgIgor Jurisica isTier I Canada Research Chair in Integrative Cancer Informatics, is a Senior Scientist at Princess Margaret Cancer Centre, Professor at the University of Toronto and Visiting Scientists at IBM's Centre for Advanced Studies. He is also an Adjunct Professor at the School of Computing, Department of Pathology and Molecular Medicine Queen's U and Department of Computer Science and Engineering at York University. Igor's research focuses on integrative computational biology and the representation, analysis and visualization of high-dimensional data to identify prognostic/predictive signatures, drug mechanism of action and in silico re-positioning of drugs. Interests include comparative analysis for mining different integrated data sets . http://www.cs.toronto.edu/~juris.All authors declare that they have no competing interests."} +{"text": "The Epidermolysis bullosa (EB) Center Freiburg is the coordination center of the German EB-Network and a national center of excellence for rare skin fragility disorders. It combines clinical activities with internationally competitive research and deals with molecular diagnostics, clinical management, elucidation of disease mechanisms, and development of evidence-based novel therapies Figure . EB is ahttp://www.eb-clinet.org], a European network of EB Centers, and the Genodermatoses Network, an international network on rare skin diseases for professionals and patients [http://www.genodermatoses-network.org] aim at establishing a European Reference Network for genetic skin diseases.The EB Center Freiburg performs molecular diagnostics, coordinates multidisciplinary care for patients and their families, advices general practitioners, medical specialists, nursing staff and therapists, and disseminates information to lay public and media. The office of the Center is available for enquiries for 24 hours and responds within 24 hours. The team includes a coordinator, physicians, nurses, a social worker, a documentary clerk, scientists and laboratory technicians with expertise in EB. The consultations are usually out-patient or day clinic appointments, but hospital admission is possible for severe cases requiring extensive medical treatments. Standardized clinical practice with a diagnostic algorithm and standardized patient documentation facilitates diagnostic processes, and a weekly EB-expert meeting evaluates all diagnoses as a quality assurance measure. Currently the EB-patient registry contains data of >1000 patients with molecular genetic diagnosis and has an associated biomaterial collection of skin biopsies, cells and blood samples. These serve as basis for research on epidemiology of EB and for clinical and laboratory investigations on novel causes, disease mechanisms, genotype-phenotype correlations and treatments for EB. - In addition to numerous international research collaborations, the EB Center is actively involved in larger structures for rare diseases. The Freiburg Center for Rare Diseases [http://www.uniklinik-freiburg.de/fzse.html] provides high-level scientific expertise, innovative diagnostics and interdisciplinary care for people with rare disorders of the skin, the musculoskeletal system, the kidney, the lung, the eye, the blood and the immune systems. Internationally, EB-CliNet ["} +{"text": "To improve our understanding of sex differences in the clinical characteristics of Parkinson\u2019s Disease, we sought to examine differences in the clinical features and disease severity of men and women with early treated Parkinson\u2019s Disease (PD) enrolled in a large-scale clinical trial.Analysis was performed of baseline data from the National Institutes of Health Exploratory Trials in Parkinson\u2019s Disease (NET-PD) Long-term Study-1, a randomized, multi-center, double-blind, placebo-controlled study of 10 grams of oral creatine/day in individuals with early, treated PD. We compared mean age at symptom onset, age at PD diagnosis, and age at randomization between men and women using t-test statistics. Sex differences in clinical features were evaluated, including: symptoms at diagnosis (motor) and symptoms at randomization .1,741 participants were enrolled . No differences were detected in mean age at PD onset, age at PD diagnosis, age at randomization, motor symptoms, or daily functioning between men and women. Differences in non-motor symptoms were observed, with women demonstrating better performance compared to men on SCOPA-COG and Symbol Digit Modality measures .Overall, men and women did not demonstrate differences in clinical motor features early in the course of PD. However, the differences observed in non-motor cognitive symptoms suggests further assessment of the influence of sex on non-motor symptoms in later stages of PD is warranted. Sex-specific differences are often manifesting in disease risk, clinical manifestations, clinical management, and prognosis in widely varied disorders ranging from cardiovascular disease to neurodegenerative disorders.\u20136 UnderlAlthough Parkinson\u2019s Disease (PD) is a common disorder with aging, sex difWe analyzed baseline data from the National Institutes of Health Exploratory Trials in Parkinson\u2019s Disease (NET-PD) Long-term Study-1 (LS-1), a randomized, multi-center, double-blind, placebo-controlled study of 10 grams of oral creatine/day in individuals who were within 5 years of PD diagnosis and who were on dopaminergic therapy for fewer than two years (early treated PD). Between March 2007 and May 2010, 1,741 subjects from 45 United States and Canadian sites were enrolled. Details of the trial design, baseline sample characteristics, and the primary trial results have been published., 20We sought to evaluate whether there are sex differences in age milestones related to disease presentation and trial randomization, including: age at symptom onset, age at PD diagnosis, age at randomization, years since symptom onset, years since PD diagnosis, and length of time between symptom onset and diagnosis. We also evaluated differences in specific clinical features: retrospective patient-reported symptoms at the time of diagnosis (motor symptoms) and symptoms present at the time of randomization .Normality of the data was tested using the Kolmogorov Smirnov test. Differences in proportions for categorical variables (demographic characteristics) between men and women were evaluated using the chi-square test or the Fisher exact test depending on assumptions. Age at symptom onset, age at PD diagnosis, age at randomization, years since symptom onset, years since PD diagnosis, and length of time between symptom onset and diagnosis were analyzed for differences in demographic characteristics between men and women; adjustment for multiple comparisons was made via Bonferroni correction with a type I error level of 0.05/6 = 0.0083. Disease presentation and randomization age milestones were analyzed using a series of t-tests for unequal variances. We compared men and women on their time from symptom onset to PD diagnosis using the Wilcoxon rank sum test because this variable was not normally distributed.Motor symptoms at diagnosis were defined by: resting tremor, rigidity, bradykinesia, postural instability, and other symptoms. To evaluate differences between men and women in motor symptoms at randomization, we compared five variables between men and women. Ambulatory capacity identifies the sum of the response to the following questions administered in the UPDRS: (i) falling, (ii) freezing, (iii) walking, (iv) gait and (v) postural stability. From the UPDRS part IV (Complications of therapy), we assessed the proportion of the waking day that dyskinesias were present with three categories of response: none, 1\u201325% of the day and >25% of the day. Due to small sample size in some categories, we collapsed the categories of 1\u201325% and >25% of the day into present (some part of the day) versus absent (none). We assessed the proportion of the waking day that the subject is \"Off\" on average with five categories of response: none, 1\u201325% of the day, 26\u201350% of the day, 51\u201375% of the day and 76\u2013100% of the day. Again, due to small sample size in some categories, we collapsed the categories of 26\u201350%, 51\u201375% and 76\u2013100% into one category of >25% of the day.We evaluated three outcomes related to clinical features: 1. Motor symptoms at the time of diagnosis and at the time of trial randomization, 2. non-motor symptoms at the time of trial randomization, and 3. daily functioning at the time of trial randomization. non-motor symptoms at the time of trial randomization between men and women using six non-motor variables: UPDRS Part I (Mentation) score, Scale for Outcome of Parkinson Disease Cognition (SCOPA-COG), Symbol Digit Modalities (SDM), Beck Depression Inventory II total score (BDI), self-reported symptomatic orthostasis, and self-reported sleep disturbance.We compared functioning at the time of randomization between men and women using seven global functioning variables: UPDRS Part II (ADL) score, Schwab and England ADL (S&E ADL), total functional capacity (TFC), Parkinson\u2019s Disease Questionnaire (PDQ-39) summary index, EuroQoL 5-D (EQ5D) utility score, Modified Rankin Scale, and care level.We compared daily Twenty three variables related to clinical features were tested for differences between men and women; therefore adjustment for multiple comparisons was made via Bonferroni correction with a type I error level of 0.05/23 = 0.002.We also evaluated differences between men and women in motor and non-motor symptoms at randomization and daily functioning at the time of randomization after adjustment for age, marital status, duration of PD, and levodopa equivalent daily dose at randomization using: (i) linear regression logistic regression ; and (iii) multinomial logistic regression .www.clinicaltrials.gov (NCT00449865). Study procedures were reviewed and approved by the institutional review board for each site, prior to enrollment of participants, who each provided written informed consent to study participation. Participating sites are listed in the Appendix.The NET-PD LS-1 trial was registered at Of the 1,741 total participants, 1,123 were male (62.5%). Men were more likely than women to be married at randomization . Other demographic characteristics were not different between men and women . We did There were no differences in the proportion of men and women demonstrating any cardinal features of PD at the time of diagnosis . The thrIn terms of non-motor features, women demonstrated better SCOPA-COG and better SDM performance compared to men . InitialPD is expected to affect more than 9 million individuals worldwide by 2030., 22 ThisUnderlying the concept of sex-related differences, it has been hypothesized that endogenous and exogenous estrogen exposure may be one of the factors involved in neuroprotection or individual symptomatic effects in Parkinson\u2019s disease., 24 The However, clinical studies of sex and PD are less clear regarding sex-related differences, reporting varied and sometimes conflicting results. For instance, some studies estimate age at symptom onset to be approximately two years later in women compared to men , 26, howOur study of over 1700 individuals with early, treated PD did not find differences between men and women in age-based disease milestones or motor features. We previously reported a small, clinically-insignificant difference in motor symptoms between men and women in this cohort, however, this difference was not present following Bonferroni correction for multiple comparisons in the current analysis.There were, however, small-magnitude differences in non-motor symptoms. Women demonstrated better cognitive performance on two measures: the Symbol Digit Modalities Test (SDMT), a screening assessment for cognitive impairment, and the SCOPA-COG, a measure of memory and learning, attention, executive function, and visuo-spatial function. The absolute magnitude of difference in cognition scores between men and women was 1.3 for SCOPA-COG and 3.1 for SDMT, which may fall below the threshold of clinical significance. It is unclear whether the difference in cognitive performance represents evidence of a sex-specific effect of PD or represents general sex differences in the performance of cognitive tests. There are conflicting reports in the literature regarding the persistence of sex-specific differences in cognition with aging., 41 The The varied results from previously published studies may relate to methodological differences in study design, such as prospective or retrospective data acquisition and differences in the study population. Sex differences may also vary throughout the course of PD. Of previously published studies, those with small samples were more likely to report sex differences in various clinical features; many of these findings equalized in larger samples. In our study of the largest clinical trial of PD patients to date, there were sex differences in non-motor manifestations, but we did not detect differences in disease onset, diagnosis, or motor symptoms. In our previous analysis, we showed that there was no difference in treatment between men and women in type of medications used or levodopa equivalent dosing.Our secondary analysis is strengthened by the large sample size and prospective acquisition of a broad battery of clinician-administered and self-report assessments, which allowed us to examine many domains of PD. The selected cognitive battery was intended to focus on cognitive deficits in PD, using a general screen (SDMT), as well as disease-specific assessments (UPDRS Part I Mentation and SCOPA-COG). However, this sample was drawn from a clinical trial designed to test the effects of a drug, not directly for epidemiological research. As a result, it is possible that our cohort of trial volunteers may not fully represent the general population of patients with PD.The difference in cognition scores between men and women may represent an important finding. Increasing cognitive impairment correlates with overall disability, and PD with dementia is associated with lower quality of life and a higher degree of caregiver burden compared to PD without dementia. Thus, thParticipating sites in the LS-1 study included: University of Alabama-Birmingham, University of South Florida, University of Southern California, Emory University School of Medicine, Oregon Health & Science University, University of Colorado, Johns Hopkins University, University of Texas Southwestern Medical Center, University of California San Francisco, University of Florida, Duke University, Louisiana State University Health Science Center-Shreveport, Michigan State University, Rush University Medical Center, University of Calgary, University of Pennsylvania, Beth Israel Deaconess Medical Center, Southern Illinois University, University of Michigan, Brigham and Women\u2019s Hospital, University of Miami, Medical University of South Carolina, Pacific Health Research and Education Institute, University of Alberta, Washington University, University of Maryland School of Medicine, University of Vermont, Northwestern University, University of Kansas Medical Center, University of Kentucky, Dartmouth Hitchcock Medical Center, SUNY Downstate Medical Center, Thomas Jefferson University, Baylor College of Medicine, Georgia Health Sciences University, Institute for Neurodegenerative Disorders-New Haven, The Parkinson\u2019s & Movement Disorder Institute-Fountain Valley, University of Virginia, Vanderbilt University Medical Center, Barrow Neurological Institute, UMDNJ Robert Wood Johnson Medical School, Malcolm Randall VA Medical Center, University of Florida-Jacksonville, Indiana University School of Medicine, and North Shore University Health System Research Institute."} +{"text": "To develop a product for people who are exposed to the particular environment or activity, their implicit need should be determined; their behaviour, activity, and task etc. can be investigated. A survey was conducted to investigate the implicit need of boating people . 32.1% ost, 2011 to December, 31st, 2014. The patents to improve thermal comfort for wearers were distinguished after considering abstract, exemplary claim, plans and International Patent Classification (IPC). IPC is made up of eight sections; A: 'Human necessities', B: 'Performing operations; transporting', C: 'Chemistry', D: 'Textiles', E: 'Fixed constructions', F: 'Mechanical engineering; lighting; heating; weapons; blasting', G: 'Physics', H: 'Electricity'. The section was subdivided into further four sublevels of 'class, subclass, group, and subgroup' [The WIPSON database was usedThe common contents of both PCT's and EU's were about the technology related to respiratory, moulding & shaping, layered products composed of specific materials, safety technology adapted for vehicle, attaching technology, shock-absorbing technology, lighting, optical parts or element, signalling, and transmission/transducer technology. The number of PCT patents for improving thermal comfort was 14 cases and the number of EU patents was 13 cases. These patents were classified into at least one of eight sections in IPC. 12 cases of PCT patents and 11 cases of EU patents belonged in section A only. A patent of PCT and a patent of EU were affiliated to section B as well as section A. A patent of PCT belonged in both sections B and F. A patent of EU was affiliated with both section A and section F. Additionally, of the subgroups considered, the patents relating to improving thermal comfort were classified into six kinds of subgroups in maximum. PCT patents tended to belong to 'Parts, details of accessories of helmets', 'Ventilation arrangement', and 'Filtering process or with filter elements' but most of EU patents belonged to 'Ventilation arrangement'. However, there were few patents in which sensor system was used but the PCT patent and the EU patents belonged to 'Electric communication technique' and 'Parts, details or accessories of helmets'/'Cushioning device' respectively.2, a compact air purifier, and a liner of breathable fabric etc. but the contents of EU patents tended to be about modifying the combination of shells or foam or transforming the original shape of shells or foam.The characteristics of PCT patents were about supplying additional parts of a helmet such as a mask of helmet with filters, a cooling/ventilating fan, shutters for ventilation, an absorber for CO"} +{"text": "The treatment of diabetes requires patient compliance, based on positive measures, inserted in social development.To compare the level of knowledge and the psychological adjustment to diabetes mellitus users of the Family Health Unit.A prospective, cross-sectional, quantitative study, with comparison groups, carried out between June-August 2012 included 207 users of the Family Health Unit Finch Low, divided into three groups according to Results of glycated hemoglobin, analyzed by standard American Diabetes Association. The sample was divided into three groups as users were diabetic treated in the unit (group A=53); newly diagnosed diabetics in the unit (group B=85) and non-diabetic patients (group C=69). The collection tools included demographic information, anthropometric and related to the disease, the questionnaire of knowledge about diabetes (DKN-A) and psychological and emotional attitudes towards disease (TA-19). As well as adherence to behaviors related to individual and social development. The variables were organized using SPSS version 17.0 software and analyzed with the Student t test, Kolmogorov-Smirnov and chi square, at 0.05 significance level.We diagnosed 41.06% of users as diabetic or pre-diabetic screening. Regardless of the group to which you belong, there was little knowledge about the disease, and negative psychological and emotional adaptation, pointing down user engagement to treatment.changes are needed in health education focused on diabetes, enabling formation of social consciousness that will motivate positive behavioral changes to patients and to society in general."} +{"text": "CDC is assisting ministries of health and working with other organizations to control and end the ongoing outbreak of Ebola virus disease (Ebola) in West Africa . The updAccording to the latest WHO update , a totalGeographic distribution of the number of Ebola cases reported during August 31\u2013September 23 indicates that recent case counts continue to be high in the areas where Liberia, Sierra Leone, and Guinea meet .Geographic distribution of the cumulative incidence of Ebola, as of September 23, indicates that the highest cumulative incidence was found in five districts in Guinea , two districts in Liberia (Loffa and Margibi), and two districts in Sierra Leone (Kailahun and Kenema) .http://www.cdc.gov/vhf/ebola/outbreaks/guinea/index.html. The most up-to-date clinical guidelines on the 2014 Ebola outbreak in West Africa are available at http://www.cdc.gov/vhf/ebola/hcp/index.html.The latest updates on the 2014 Ebola outbreak in West Africa, including case counts, are available at"} +{"text": "Technology is increasingly being used in medical education to supplement the delivery of learning resources. Gamification is defined as \u2018the use of game design elements in non-game contexts\u2019 . DespiteA systematic literature review conducted by Hamari et al. has showThere are many examples of the successful implementation of gamification in a variety of fields. For example, American Airlines offering rewards in their frequent-flyer programme in 1982, or more recently Foursquare, a location-based social network, using gamification successfully to reward users for \u2018checking-in\u2019 . Foldit,Social Media Innovation course at the Fox School of Business at Temple University. Steven L. Johnson gamified the learning process with a mission-based narrative where students leveled up after a certain number of accomplishments, such as achievements or badges (Just Press Play initiative at the Rochester Institute of Technology. Game elements and a narrative outline were used to create an educational game that promoted academic success (There are also examples of successful use cases in the university setting, such as the gamification of the r badges . Another success .In the last decade, there has been increasing interest in medical educational strategies, with the development of new concepts such as problem-based learning, student-centered learning, and integrated teaching. Many theories exist regarding the next move for medical education. Given the success of gamification in other educational settings, the implementation of game elements in medical education could provide an innovative solution. Further research is needed to evaluate the effectiveness of gamification in medical education.Maroof Ahmed Faculty of Medicine, Imperial College London, London, UK Email: maroof.ahmed11@imperial.ac.ukYusuf Sherwani Faculty of Medicine, Imperial College London, London, UKOsama Al-Jibury Faculty of Medicine, Imperial College London, London, UKMuhammad Najim Faculty of Medicine, Imperial College London, London, UKRiham Rabee Faculty of Medicine, Imperial College London, London, UKMuhammad Ashraf Faculty of Medicine, Imperial College London, London, UK"} +{"text": "The Funding section is incorrect. The complete, correct Funding statement is: TCP acknowledges support of the MIDAS study NIH NIGMS U01-GM087728. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The International Workshop on Semiconductor Characterization: Present Status and Future Needs was held at the National Institute of Standards and Technology from January 30 to February 2, 1995. This comprehensive, \u201cworld-class\u201d workshop was dedicated to summarizing major issues and giving critical reviews of important semiconductor characterization techniques that are useful to the semiconductor industry. Because of the increasing importance of in-line and in-situ characterization methods, the workshop placed a strong emphasis on these methods.Specific goals of the workshop were: (1) to provide a forum in which measurements and technical issues of current and future interest to the semiconductor industry could be reviewed, discussed, critiqued, and summarized; (2) to demonstrate and review important applications for diagnostics, manufacturing, and in-situ monitoring and control in real-time environments; (3) to provide a silicon integrated circuit process and materials-based view of requirements for off-line, in-line, and in-situ analysis and metrology; (4) to focus attention on the critical and unique requirements related to compound semiconductor materials and devices; and (5) to act as an important stimulus for new progress in the field by providing new perspectives.The workshop provided a concise and effective portrayal of industry characterization needs and the problems that must be addressed by industry, government, and academia to continue the dramatic progress in semiconductor technology.Semiconductors and their applications are one of the greatest scientific and technological breakthroughs of this century. Consider the impact of microelectronic components used in computers, entertainment equipment, automotive electronics, medical instrumentation, telecommunications, space technology, television, radio, and manufacturing technologies. Almost every factory, hospital, office, bank, school, or household contains transistors, microprocessors, and other semiconductor devices. Semiconductor characterization is an indispensable enabler of all modern microelectronics and optoelectronic circuits, and is in the critical path for maintaining the steady decline in cost-per-function of silicon integrated circuit technology. It is also helping to drive new developments in compound semiconductor materials and devices (III\u2013V and II\u2013VI).Semiconductor materials and devices must continually meet more stringent requirements as the density and performance of semiconductor devices increase. The purity, perfection, and cleanliness required of the materials; the ultra-small dimensions of the devices; and the device properties themselves require measurements to a higher precision and with a resolution and sensitivity that pushes these techniques to their very limit. In addition, new techniques associated with analysis of process chemicals, control of process steps, and characterization of packages are critically needed. Because the materials and devices themselves exhibit a rich variety of properties, an increasingly wide range of measurement techniques has evolved to meet industry\u2019s needs. Meeting the demands for large-scale, complex, integrated circuits will continue to require technological advances in materials, processing, circuit design, characterization, testing, and standards.Compound semiconductors are also being used in a variety of structures for light-emitting diodes, laser diodes, far-infrared detectors, and microwave devices. In addition to being used in their bulk and natural forms, these semiconductor materials are used in artificially created structures such as superlattices and heterostructures. Various compounds are mixed to produce structures in which properties such as the bandgap have been engineered to have specific values. Such structures and devices have unique applications in wireless communications, optical communications, visible light sources, and imaging, all of which are critical for information technologies. They also enable functions and enhanced performance that cannot be equaled by silicon-based technologies.1992 SIA Roadmap. Here, key industry, government, and university technologists described a strategy for meeting materials, process, tool, and factory requirements for future IC manufacturing. The document was updated 2 years later as the National Technology Roadmap for Semiconductors (NTRS) [International economic competition is pressing the U.S. semiconductor industry to promote precompetitive cooperation among government, industry, and universities. A unified viewpoint fosters efficient use of government and university resources and effective planning for future activities. In 1992, the Semiconductor Industry Association (SIA) published a consensus view of the requirements for the manufacture of future silicon-based integrated circuits in a document known as the A good description of materials characterization considers it as an integral part of process development and manufacturing. The Office of Science and Technology Policy 1993 fiscal year program booklet describes materials characterization as a wide range of interdisciplinary activities that determine the structure, composition, properties, and performance of materials, and the relationships among these . Charactmetrology. Engineers often use the word to describe procedures, such as critical-dimension (CD) measurements, which routinely monitor lithography processes inside the clean room. Others generalize it to all in-line measurements. According to the dictionary, metrology is the science of measurement [characterization is also sometimes used interchangeably with metrology.The semiconductor industry refers to measurements used in process control as surement . It shousurement . The ternon-value-added activity. For in-situ real-time control, it can improve equipment effectiveness by reducing process time, down time, and use of test wafers, while maintaining product quality. In some cases, it is advantageous to replace metrology tools requiring test wafers with others that measure product wafers nonintrusively. In many wafer fabrication facilities (fabs), defect metrology is a key element in maintaining high yield. This includes particle and defect detection and characterization on patterned wafers using particle sensors and electrical yield testing.Metrology must break the status quo image of being a In high-yield manufacturing, metrology requirements switch from fundamental measurement to verification of repetitive execution. In-situ and in-line sensors are evolving for process control, and today reach the manufacturing floor through vendors who integrate them into new equipment. Sensors add value by detecting excursions early on that might persist through the wafer fab with cumulative line yield loss.Analytical laboratory researchers sensitive to this perspective focus their efforts on technologies that are less mature. In contrast, process development engineers sometimes stretch existing metrology too thin by pushing sophisticated instruments into a regime where they are not very effective or technically applicable. Metrology has the potential to establish a common denominator for transitions between all of the process maturity phases, and thereby reduce operational barriers.Metrology tools applied in-line to manufacturing, and in-situ inside commercial instruments are initially developed as scientific prototypes in the R&D environment. These then evolve into stable user-friendly modules, suitable for introduction into the off-line wafer fab analytical laboratory. Further maturity brings durability and reproducibility through robust design adaptable to the wafer fab environment. Specifications and standard test methods developed by organizations such as SEMI and ASTM, and the availability of applicable certified reference materials such as NIST Standard Reference Materials (SRMs) facilitate successful transfer through these three stages. Training in relevant materials science as well as procedures of calibration are also important.In-situ measurements are not common in volume manufacturing in the semiconductor industry today. The NTRS predicts that required factory equipment efficiency improvements will drive future incorporation of in-situ metrology. As fab managers realize the impact of a measurement tool on throughput, they will drive equipment vendors to incorporate such in-situ sensors. Ellipsometry is an example of a metrology that has progressed from R&D laboratory to an in-situ process control tool. One quarter of a century elapsed between the first R&D application of ellipsometry to semiconductor dielectric films and the demonstration of an in-situ sensor.The progressive shrinking of ultra-large-scale integration (ULSI) circuits into the submicrometer regime and the emergence of quantum structures on the nanometer scale increase the challenge for characterization specialists. When the probe radius, depth of analysis, or contaminant level is reduced, the volume of analysis ultimately includes only several atoms of interest. In the extreme, the volume analyzed contains no atoms at all. The analyst is now confronted with a sampling problem in selecting which atoms are appropriate for the analysis. Single atom detection cannot be considered independently of the sampling problem. This issue is becoming more visible, as we realize a related problem is looming, not just in characterization, but also in the fabrication of lightly doped 3-D shallow junctions of nanometer dimensions, where only a few, or even single, dopant atoms may someday be required.The NTRS established the groundwork for the creation of a metrology roadmap for silicon, which was developed through SEMATECH . A compaMaterials parameters generic to III\u2013V compound semiconductors include bandgap energy, band offsets, interface diffusion and roughness, index of refraction and index dispersion, optical absorption, minority and majority carrier lifetime and mobility, and defects that influence lifetime or trapping of charge. Contactless tools include ellipsometry, light scattering, optical reflectometry, and photoreflectance, which are applied to heterostructures grown by molecular beam epitaxy (MBE) and other advanced thin-film techniques, e.g., metalorganic chemical vapor deposition (MOCVD) and metalorganic-MBE (MOMBE). These optical measurements are by nature noninvasive, and are quickly finding niches as in-situ process sensors that permit optimization of film thickness, composition, interface quality, and uniformity during growth.2. Recent advances in surface passivation, liquid phase epitaxial (LPE) growth, and device processing have resolved many of the major materials problems encountered in the past. The principal limitation is now control of crystal point defects and impurities, especially for devices operating at 78 K or lower temperatures. There is critical demand for smaller analytical probes with higher sensitivity to resolve such problems. Incomplete understanding of physical and chemical defects continues to inhibit progress with basic II\u2013VI materials issues.Infrared detectors and imagers based on II\u2013VI compound semiconductors are key enablers in a variety of military and space applications. HgCdTe is the material of choice for both scanning and staring focal-plane arrays. The complexity of ternary alloy systems and the stringent demands of large element detector arrays combine to increase the difficulty in working with II-VI compounds by at least an order of magnitude relative to GaAs. Material quality has reached the level of maturity required for two-dimensional focal-plane arrays based on epitaxially grown HgCdTe wafers as large as 30 cmCompound semiconductors are also candidates for high-temperature electronics applications. Although diamond offers high-thermal stability and mechanical hardness for power transistors, difficulty in growing thin films with suitable lattice-matched substrates is making it less attractive to low-cost commercial markets. Active research with compounds such as SiC and GaN is in progress. GaN has added advantages for light-emitting display applications. The characterization tools outlined above are also applicable to these developments.cellular automaton). Material systems are typically heterostructures composed of lattice-matched and doped GaAs/AlGaAs and related compounds.Nanoelectronics is a revolutionary digital IC technology directed at continued downsizing of minimum feature size below 0.1 \u03bcm. Priority is given to eliminating long interconnects, because these are the most difficult to scale down. The approach leads to an architecture in which resonant electron quantum tunneling effects dominate, and each active element is connected only to its nearest neighbor cells to provide for multiple inputs and interactive discussion, with emphasis on the measurement equipment supplier perspective and on important issues related to the topics of the invited paper sessions. A special evening rump session explored the potential and promise of synchrotron x-ray metrology for TXRF and other analytical techniques.The workshop was organized by a program committee chaired by David G. Seiler, National Institute of Standards and Technology. Other members of the committee were: David E. Aspnes, North Carolina State University; Ray Balcerak, Advanced Research Projects Agency; W. Murray Bullis, Materials & Metrology; Alain Diebold, SEMATECH, Inc.; Wolfgang Jantz, Fraunhofer-Institut f\u00fcr Angewandte Festk\u00f6rperphysik; Alan Jung, SEMI; Sanjiv Kamath, Hughes Research Laboratories; Stephen S. Laderman, Hewlett Packard; Bob McDonald, Intel Corporation; William T. Oosterhuis, U.S. Department of Energy; Abbas Ourmazd, AT&T Bell Labs; Paul S. Peercy, Sandia National Laboratories; Fred H. Pollak, Brooklyn College of SUNY; John Prater, Army Research Office; Tom Remmel, Motorola; Linton G. Salmon, National Science Foundation; Tom J. Shaffner, Texas Instruments; Richard A. Singer, Institute for Defense Analyses; and William E. Tennant, Rockwell International Science Center. A NIST Advisory Committee consisting of Paul M. Amirtharaj, Frank F. Oettinger, Robert I. Scace, and James R. Whetstone also assisted in the organization of the workshop.Chairs of the invited paper sessions were Dirk Bartelink, Hewlett-Packard; James Freedman, SRC; Len Feldman, AT&T Bell Labs; Stephanie Butler, Texas Instruments; Anne Testoni, Digital Equipment Corporation; P. B. Ghate, Texas Instruments; Paul Ho, University of Texas; Richard Brundle, Brundle Associates; Richard S. Hockett, Charles Evans & Associates; David E. Aspnes, North Carolina State University; C. Pickering, Defence Research Agency; John Prater, Army Research Office; Fred H. Pollak, Brooklyn College of SUNY; John Parsey, Motorola, Inc.; and Jerry Woodall, Purdue University. Mike Fossey, ADE Corporation; Robert I. Scace, NIST; and John C. Bean, AT&T Bell Labs chaired the panel sessions.Sponsors of the workshop were: The Advanced Research Projects Agency, SEMATECH, the National Institute of Standards and Technology, the Army Research Office, the U.S. Department of Energy, the National Science Foundation, SEMI, the Manufacturing Science and Technology Division of the American Vacuum Society, and the Working Group on Electronic Materials of the Committee on Civilian Industrial Technologies.In an evaluation survey conducted following the workshop, respondents indicated that the workshop had been meaningful and relevant for them because it (1) gave them insights and priorities rather than simple listings, (2) provided useful information of the capabilities of different characterization techniques, (3) gave perspectives on industrial metrology requirements, (4) explored critical needs and issues in semiconductor metrology research, (5) was relevant to technologists, and (6) brought together the international metrology community at NIST, a logical place to hold the workshop. There was considerable interest in holding another similar workshop in 2 years to 3 years in order to cover emerging techniques in semiconductor characterization as this important field develops further.The proceedings volume, published by the American Institute of Physics, is intended to serve as a base-line reference for the characterization of semiconductors for the next decade . It begiThe remainder of the volume is organized along the lines of the workshop program. The papers are grouped in seven major sections under the topics of the invited paper sessions: Drivers for Silicon Process Development and Manufacturing; Metrology Requirements for beyond 0.35 \u03bcm Geometries; Silicon Materials, Gate Dielectrics, and Process Simulation; Interconnects and Failure Analysis; Critical Analytical Methods; In Situ, Real-Time Diagnosis, Analysis, and Control; and Frontiers in Compound Semiconductors. The section on Critical Analytical Methods is further divided into eight sub-sections according to category of method. Brief summaries of the panel and rump sessions and the after-dinner remarks on Historical Perspective on Semiconductor Characterization are included as appendices."} +{"text": "Living in green/blue areas is associated with better health. This may be due to low air and/or noise pollution, opportunities for physical activity, facilitation of social contacts, and promotion of recovery from fatigue and stress. Yet, socio-economic (SE) factors also explain inequalities in health and access to green/blue spaces. The GRESP-HEALTH project evaluates the associations between living in/close to a green/blue area on morbidity and mortality in Belgium. It assesses all-cause and cause-specific mortality, specific morbidities and perceived health, considering environmental pollutants and SE factors.The project includes individuals registered in the Belgian censuses of 1991 and/or 2001. Three levels of observation are studied: individual, statistical sector (SS) and group of SS, following individual and ecological designs. Mortality information is based on the National Mortality Database , perceived health (2001 census) and SE factors (1991 and 2001 censuses). Morbidity information (2004-2012) is derived from the IMA database, which contains reimbursement data of prescriptions. For green/blue spaces, the surface, shape, accessibility and type are calculated for each SS. Residential area-specific exposure to air pollutants is obtained from satellite images. Traffic noise databases are used whenever possible. We will consider SE factors such as material deprivation, education, and occupation. The analyses will be conducted separately in different age specific populations and types of area . We will use multilevel models for clustered data within geographical areas. Interactions of green/blue spaces with air pollution and SE factors will be evaluated and stratified analyses in areas with similar SE and environmental characteristics will be performed. Moreover, specific population groups will be considered.The GRESP-HEALTH project will improve the scientific knowledge about the hitherto uncertain associations between living close to green/blue spaces and health."} +{"text": "Aboriginal Populations: Social, Demographic, and Epidemiological Perspectives. Frank Trovato & Anatole Romaniuk, editors. Edmonton, Canada: University of Alberta Press, ISBN-13: 978-0-88864-625-5Experts from around the world review and extend the research on Aboriginal peoples in Canada, Australia, New Zealand, and the circumpolar North, mapping recent changes in their demography, health, and sociology and comparing their conditions with that of Aboriginal peoples in other countries.Contributors point to policies and research needed to meet the challenges Aboriginal peoples are likely to face in the twenty-first century. This substantial volume will prove indispensable and timely to researchers, policy analysts, students, and teachers of social demography and Native Studies.orders@gtwcanada.comOrder from:"} +{"text": "The National Computer Systems Laboratory (NCSL) of the National Institute of Standards and Technology (NIST) and the National Computer Security Center (NCSC) of the Department of Defense (DoD) co-sponsored the Twelfth National Computer Security Conference held in Baltimore, Maryland on October 10\u201313, 1989. The theme of the conference, \u201cInformation Systems Security: Solutions for Today\u2014Concepts for Tomorrow,\u201d highlighted the broader focus of information systems security which now challenges the user community, vendors, system developers, and administrators. Major areas addressed included advanced research developments and emerging technologies, network security architectures, risk management, management and administration issues in computer security, and an expanded focus on education and ethics. More than 2,000 attendees from government, industry, and academia participated in the 4-day conference, which was co-chaired by NCSL\u2019s Irene Gilbert and NCSC\u2019s George Mitchell.The National Computer Systems Laboratory has played a vital role in protecting the security and integrity of information in government computer systems through its Computer Security Program since the passage of the Brooks Act in 1965. Since 1972, the Laboratory has issued standards and guidelines on the cost-effective protection of unclassified information in computer systems. NCSL works with organizations from government, industry, academia, and voluntary standards groups to develop standards and guidance, produce tests to evaluate conformance to standards, transfer technology to users, and provide technical assistance to government and industry in computer security applications. The Computer Security Act of 1987 strengthened NCSL\u2019s role in protecting unclassified sensitive information in federal computer systems.The Department of Defense also has pursued an active computer security program over many years. In 1978, the Assistant Secretary of Defense for Communications, Command, Control and Intelligence established a Computer Security Initiative to ensure the widespread availability of trusted ADP systems within the DoD community. In 1981, the National Computer Security Center was created to administer the activities of the initiative. NCSC advances the development of trusted computer systems and publishes guidelines on computer security. In response to the increased emphasis on computer security in recent years, NCSC expanded its efforts to support basic research for the development of additional trusted systems. NCSC\u2019s technology transfer program ensures that other federal agencies and industry benefit from technological advances in computer security.Sharing common goals and mutual challenges, NCSL and NCSC joined forces in 1979 to co-sponsor the first National Computer Security Conference. Dr. Dennis Branstad, NCSL (now a NIST Fellow) and Mr. Stephen Walker, then Chairman, Computer Security Technical Consortium, organized the first seminar as an information exchange on computer security issues. Since its inception, the meeting has expanded significantly in size and scope to become a large, comprehensive computer security conference. This development parallels an increased national awareness of the need for computer systems security and an expanded interest in existing and emerging technologies available to protect vital information resources.The Twelfth National Computer Security Conference was organized into four tracks: research and development; systems; management and administration; and education and ethics. Of particular interest was the expanded focus on computer security education and awareness resulting from requirements of the Computer Security Act of 1987 and the in-depth concentration on computer security ethics in the workplace. The first-day \u201cOverture\u201d sessions offered an introduction to basic computer security subjects including an overview of agency security plans submitted in response to the Computer Security Act, ethical conflicts in computer science, NCSL/National Security Agency (NSA) joint efforts, the Secure Data Network System, DoD Trusted Computer System Evaluation Criteria, and training guidelines.The second day of the conference opened with welcoming remarks from NCSC Director Patrick R. Gallagher and NCSL Director James H. Burrows. Representative Tim Valentine of North Carolina presented the keynote address on the role of Congress in computer security. Valentine cited the progress of federal agencies in implementing the Computer Security Act and called for expediting federal progress in computer security technology and standards, including international standards. An opening plenary on \u201cInformation Systems Security\u2014A Year in Review,\u201d followed. Participants included James Burrows, NCSL; Patrick Gallagher, NCSC; Clive Blatchford, ICL, United Kingdom; Steve Kent, Bolt, Baranek, and Newman; Stephen Walker, Trusted Information Systems; Stuart Katzke, NCSL; Eliot Sohmer, NCSC; and Harold Segal, Office of Personnel Management. Among the subjects addressed were legislation and policy; the international standardization effort; electronic data interchange; trusted systems, virus response centers; international trusted criteria; and computer security training and awareness.Following this overview, speakers from government and industry presented concurrent sessions in the four tracks described above. Selected presentations are summarized below.Achieving multilevel database security was the focus of the database management session chaired by Teresa F. Lunt of SRI International. Gary W. Smith of George Mason University presented a paper describing a balanced approach which uses good system design, management controls, and procedural security as well as technical solutions to achieve multilevel database security.Tim Wood of Sybase, Inc. gave an overview of the system architecture of the Sybase Trusted SQL Server, targeted at the B2 level of trust. The Trusted SQL Server is a physical machine control program that is a hybrid of a secure, high-performance DBMS server with a dedicated kernel of original design. A third paper by R. Alan Whitehurst, University of Illinois at Urbana-Champaign, and Teresa F. Lunt, SRI International, discussed the verification of the SeaView formal top-level specifications and the benefits gained from formally specifying and verifying selected database operations. The SeaView project was a 3-year program to create the design of a multilevel secure relational database system that met the criteria for trusted system Class Al.Joshua Guttman, MITRE Corporation, chaired this session. Carla Marceau and C. Douglas Harper, Odyssey Research Associates, presented a paper describing an interactive approach to Ada verification. Penelope is a prototype Ada verification editor whose user interactively and concurrently develops specifications of programs, their Ada text, and proofs of their verification conditions. Adding CASE technologies to formal verification was the subject of the next paper by J. V. A. Janeri, J. S. Barlas, and L. L. Chang of the MITRE Corporation. The authors used CASE technology to further automate the labor-intensive task of formal verification, by integrating the process of formal software design verification with the software engineering life cycle. The result is a Specification Browser which serves as a verification aid. In the final paper of this series, Timothy E. Levin, Steven J. Padilla, and Roger R. Schell, Gemini Computers, Inc., presented engineering results from the trusted system level Al formal verification process. The authors reviewed the formal methodology used to verify the security of the GEMSOS TCB, currently under development and targeted for the TCSEC Class Al level.Marvin Schaefer, Trusted Information Systems, chaired the verification session. Barbara A. Mayer and Monica M. McGill, NCSC, presented an overview and rationale of recently published guidelines for formal verification systems. The paper described the history and status of the guidelines, the endorsement process, the evaluation approach, and possible future directions for verification systems.A second paper by William D. Young, Computational Logic, Inc., compared and contrasted the Gypsy and Z specification languages. The authors suggested refinements to the two languages and pointed a direction for future language designs. The final paper in this group evaluated security model rule bases; John Page, Jody Heaney, Marc Adkins, and Gary Dolsen, Planning Research Corporation, are the authors of the paper. The evaluation viewed three different security models from the common point of reference provided by the Security Model Development Environment prototype.The session on models was chaired by D. Elliot Bell, Trusted Information Systems. In the first presentation, Robert S. Lubarsky, Franklin and Marshall College, described a mathematical approach to hook-up security and generalized restrictiveness. A second presentation explained the Argus computer security model.NCSL\u2019s Lisa Carnahan and NCSC\u2019s Mario Tinto chaired these sessions. Papers focused on a broad range of products and systems: the design of trusted workstations using a total \u201cinformation security\u201d (INFOSEC) solution; Formal Top Level Specification (FTLS) security testing for the Honeywell LOCK project; the formal specification of security aspects of a messaging system architecture; a Secure Distributed Operating System (SDOS) prototype; a high B level security architecture for the IBM ES/3090 Processor Resource Systems Manager; and a TRW security engineering effort to define an architecture for a MLS communications processor. An R&D panel and an ethics plenary session concluded the track.Network security architectures received in-depth coverage, focusing on privacy and access control issues. Recent incidents involving malicious code have drawn increased attention to the need for prevention and remedies in this area.Ruth Nelson, GTE, chaired this session. John Linn, DEC, and Stephen Kent, BBN Communications Corporation, presented a paper on privacy for DARPA-Internet mail. The facilities discussed provide privacy enhancements on an end-to-end basis between originator and recipient User Agent processes, which may be implemented on heterogeneous systems. The authors defined and recommended a cryptographic key management approach employing RSA-based public-key certificates.Key management and access control for an electronic mail system was the subject of the next paper by Martha Branstad, W. Curtis Barker, Pamela Cochrane, and David Balenson of Trusted Information Systems, Inc. The authors examined key management and access control services associated with the Embedded Network Security (ENS) Trusted Mail system, indicating how both encryption and trusted system functionality provide protection. Miles Smid, James Dray, and Robert Warnar, NCSL, described a token-based access control system for computer networks. In this system, a user\u2019s access is mediated by a smart token implementing a transparent cryptographic three-way handshake with the target computer.The increasing use of local area networks (LANs) has driven the search for cost-effective security solutions. Dennis Branstad, NCSL, chaired the session on LANs. Gary Stoneburner and Dean Snow, Boeing Aerospace and Electronics, described how and why the Boeing Multilevel Secure local area network (MLS LAN) is migrating towards an Information Security (INFOSEC) solution. Significant design issues were presented, as well as an overview of how encryption might be embedded into the MLS LAN. L. Kirk Barker, Datotek, and Kimberly Kirkpatrick, MITRE, next described the Standard for Interoperable LAN Security (SILS) model which would provide a standard protocol for protecting LAN traffic; the IEEE 802.10 is basing its security protocols and services for LANs on this model. Peter Loscocco, NCSC, presented the last paper in this series on a dynamic network labeling scheme for a MLS LAN.Donna Dodson, NCSL, chaired the networks session addressing access control; Dennis Grayson, NCSC, chaired the second networks session. Extending mandatory access controls to a networked multilevel secure (MLS) environment was the subject of the first presentation by Ron Arbo, Eric Johnson, and Ron Sharp, AT&T Bell Laboratories. They introduced a software package design that permits MLS systems to securely communicate without modifying or trusting the existing network applications. Richard Graubart, MITRE, reexamined the traditional access control policies and proposed a new type of access control policy. Four DEC researchers next described a digital distributed system security architecture. Other papers in this series outlined guidelines for specifying security guards and the security of embedded tactical systems.Protecting information systems from threats of all kinds was the focus of these sessions; Jack Holleran, NCSC, and James Anderson, J. P. Anderson Co., were session chairs. Martha Brothers, AT&T, gave a \u201chow to\u201d guide for virus protection in MS-DOS, while Ronald Tencati, Goddard Space Flight Center, and Patricia Sisson, Science Applications Research, described the \u201cFather Christmas Worm\u201d of December 1988 which invaded a large DECnet network and reached 6,000 computer nodes worldwide. Cliff Stoll, Harvard\u2014Smithsonian Center for Astrophysics, gave an epidemiology of viruses and network worms. Other talks summarized computer crime techniques and gave potential solutions.The systems track concluded with a discussion of vendor activities by Dennis Sirbaugh, NCSC, and a plenary session on ethics.Irene Gilbert was session chair for management. William Norvell, Hughes Aircraft Company, gave the first presentation on integrating accreditation activities into the acquisition process, ensuring that all security requirements are specified in the functional baseline for design and test. David Juitt, Digital Equipment Corporation (DEC), proposed a security approach through system management; he detailed an ongoing advanced development effort within DEC to study security issues of computing across a worldwide distributed environment and how they relate to conducting business safely. A third paper described a systematic approach to software security evaluations.In this session chaired by Grant Wagner, NCSC, the first presentation by Toni Fish, Information Systems Security Association, and Corey Schou, Idaho State University, addressed the issue of the certification of computer security professionals. Darryl Song, MEI Associates, spoke on the accreditation of information systems and networks, followed by a talk by Horace Peele, Electronic Security Command (ESC), USAF, on the development of the ESC Accreditation Package. Peele recommended automated accreditation packages as effective security tools throughout the Department of Defense and the federal government.These sessions were led by Sylvan Pinsky, NCSC, and Irene Gilbert, NCSL. Jennie Stevens and Richard Weiner, Booz, Allen & Hamilton, Inc., presented an innovative concept for computer security risk assessment developed in 1988 and 1989 by their company. The concept provides a framework upon which an individual organization\u2019s customized guideline can be built. Next, Suzanne Smith, Los Alamos National Laboratory, introduced LAVA, the Los Alamos Vulnerability/Risk Assessment system, a three-part systemic approach to risk management that can be used to model a variety of application systems. A third paper focused on the purpose and framework of anomaly detection; G. Liepins, Oak Ridge National Laboratory, and H. Vaccaro, Los Alamos National Laboratory, placed anomaly detection of computer use in the framework of overall computer security. NCSL\u2019s Stuart Katzke gave an update of progress in the federal government\u2019s risk management activities.The remainder of Track C dealt with Air Force customer programs, a \u201cspeak out\u201d session for the informal exchange of ideas and opinions, an outline of NCSL programs by Miles Smid, and a concluding plenary session on ethics.This new track on computer security education and ethics attracted much interest.Charles Pfleeger, Trusted Information Systems, chaired the overview session on ethics. In the first presentation, Larry Martin, Subcommittee on Automated Information Systems Security, addressed the issue of responsibility for unethical computer behavior. Martin concluded that while the ultimate responsibility to behave in an acceptable manner belongs to the user, all of us share in the responsibility for and the consequences of unethical computer use. Glenn D. Watt, NCSC, spoke next on the ethical dilemma of malicious code; his solution combines technology and ethics. A panel discussion followed.Management responsibility versus individual rights was the focus of the next session, chaired by Lance Hoffman, George Washington University. The first speaker was Robert Veeder, Office of Management and Budget, whose topic was the Computer Matching and Privacy Protection Act of 1988. Anna Patrick, U.S. Department of Agriculture, spoke next on public access to government databases. A panel discussion followed on the role of management versus the prerogatives of individuals; panel members included session speakers plus Brian Hyland, U.S. Department of Labor; Jan Goldman, ACLU; and Marc Rotenberg, Computer Professionals for Social Responsibility.William Murray, Ernst & Whinney, chaired the next session on the criminalization of computer abuse and misuse. Jay BloomBecker, National Institute on Computer Crime Data, discussed trends in computer misuse, followed by James Miller, University of Southern Mississippi, who gave an academic perspective on computer abuse. A prosecutor\u2019s point of view was presented by William Cook, U.S. Attorney\u2019s Office; he successfully prosecuted the first case under the Computer Fraud and Abuse Act of 1986. A panel discussion concluded the session.Who is responsible for ethics in the workplace? James P. Anderson, James P. Anderson Co., led the session addressing this issue. Karen Forcht, James Madison University, presented a talk on the ethical use of computers, followed by a panel discussion.At this point, the focus of Track D shifted to education, training, and awareness issues in computer security. Lauresa Stillwell, Department of State, chaired the session. W. V. Maconachy, NSA, discussed turning a philosophical orientation of computer security education into a practical reality. He challenged government personnel to develop and implement a well-orchestrated government-wide information systems security awareness, training, and education model. A panel then compared and contrasted the education, training, and awareness continuum, followed by a talk by John Higgins, Brigham Young University, on training computer science undergraduates in information security.Harold Segal, Office of Personnel Management (OPM), led the session on computer security training in the federal government. Following his talk on OPM training modules, Anne Todd, NCSL, presented the training guidelines which she coauthored. A federal training panel concluded the session.The computer security awareness session was chaired by Harry DeMaio, Deloitte, Haskins & Sells. His talk on employee awareness was followed by one on executive awareness by Joan Foreman, Bureau of Engraving and Printing. An executive awareness panel preceded the closing talk on mandated versus voluntary ethics given by Marlene Campbell, Murray State University (Kentucky).The Twelfth National Computer Security Conference covered a broad range of issues and emerging technologies of value to those charged with the responsibility of protecting vital information resources in computer systems. \u201cInformation Systems Security: Solutions for Today\u2014Concepts for Tomorrow\u201d offered guidance for the present and planning strategies for the future. In the rapidly changing world of information systems technology, the importance of planning for the security of tomorrow\u2019s information systems is critical to all who understand that an organization\u2019s information is its most valuable asset.Conference proceedings are available from conference co-chair Irene Gilbert, National Computer Systems Laboratory, A216 Technology Building, National Institute of Standards and Technology, Gaithersburg, MD 20899, or you may call (301) 975-3360."} +{"text": "In 2008, the Global Health Research Initiative (GHRI) invited applications from teams of researchers and decision-makers who were interested in conducting research related to human resources for health and the implementation and use of integrated health information systems in Africa, with special attention to equity considerations. These thematic areas constituted the focus of the Africa Health Systems Initiative - Support to African Research Partnerships (AHSI-RES) program.The Global Health Research Initiative is a partnership of three Canadian agencies: Foreign Affairs, Trade and Development Canada (DFATD), International Development Research Centre (IDRC), and the Canadian Institutes of Health Research (CIHR). GHRI is hosted at IDRC. AHSI-RES was a five year, $5.9 million CDN research program (2008-2013) supported by Foreign Affairs, Trade and Development Canada ($5 million) and the International Development Research Centre ($900 000). AHSI-RES is the research component of the larger DFATD Africa Health Systems Initiative (AHSI) program. The AHSI program is a 10 year, $450-million CDN commitment (2006-2016) to strengthening national-level health strategies and architecture, and is being implemented by Foreign Affairs, Trade and Development Canada.The AHSI-RES program\u2019s purpose is to support policy relevant research, knowledge translation and exchange in the program\u2019s thematic areas. The AHSI-RES program emphasized the importance of ongoing interaction, collaboration, and exchange of ideas between researchers and decision-makers to maximize the likelihood that research findings would be used to inform programs and policies. A decision-maker was defined as \u2018an individual who makes decisions about, or influences, health policies or practices.\u2019 The program used different approaches in order to build or increase local capacity for research, knowledge translation, and research use. The long-term objective was: \u201cHealth systems research allows African decision makers, policy advocates and health service managers to improve health outcomes and reduce disease burden through more efficient and affordable health systems\u201d .Teams were required to include one African researcher and one African decision-maker, both as co-principal applicants. Other African and non-African researchers and decision-makers could be involved as co-applicants or as collaborators. The co-principal applicants had to be affiliated with an institution located in an AHSI-RES geographic area of focus. Geographic areas of focus included: Francophone West Africa ; Great Lakes and Eastern Africa ; and Southern Africa .Ten teams were selected from fifty-seven proposals following a rigorous merit review process, which included both researchers and decision-makers. Successful teams had to demonstrate: the ability to engage in interdisciplinary applied research to address complex policy-relevant questions related to the key themes of AHSI-RES; and, the ability to link research, policy, and action to improve health decision-making and programming. Gender, ethics, capacity development, and knowledge translation and exchange were cross-cutting themes featured in AHSI-RES programming.BMC Health Services Research. The AHSI-RES program also produced important results which aim to address the human resources for health crisis through task-shifting, recruitment, and retention. The Human Resources for Health journal, due to the relevance of its thematic focus, was selected for the publication of this supplement.Selected teams were from seven African countries: Burkina Faso, Mali, Kenya, Tanzania, Uganda, Malawi, and Zambia. Their research focused on two main areas: 1) the recruitment and retention of health workers and the shifting of certain tasks to less specialized health workers (i.e. task-shifting) in response to a severe human resources crisis in the health sector in sub-Saharan Africa; and 2) the role of health information in ensuring greater equity in access to health care. The lessons learned from the uptake and impact of AHSI-RES research for evidence-based practice is presented in a supplement to The ten teams supported by this program worked to connect research with policy and action to improve health decision-making and programming in the sub-Saharan region, paying particular attention to the needs of disadvantaged segments of the population. Human resources for health are especially scarce in certain specialties and in certain regions, mainly those which are furthest from urban centres. Understanding how to successfully recruit trained health personnel, maintain their motivation, and retain them in these rural and remote areas has the potential to expand access to health services to the most vulnerable and strengthen health systems in sub-Saharan Africa, a region at the centre of the human resources for health crisis.The articles in this supplement present research results from Burkina Faso, Tanzania, Uganda, and Zambia.A main focus of the project teams is task-shifting, though research on this human resources strategy spans diverse fields including mental health, surgery, community health, and ophthalmology. Primary health care workers provide primary eye care services in Kenya, Tanzania, Malawi, and Madagascar. However, as research described in this supplement shows, their competency scores are low , as are Another team studied job descriptions in Uganda, with the goal of understanding the current scope of the mandates of cadres of health care workers who provide surgical services . These wRecruitment, motivation, and retention have also been major foci of the AHSI-RES program research teams. Although different recruitment and retention strategies have been put in place by national governments, the effectiveness of these strategies is not often evaluated. One team chose to explore this in Zambia. Despite studying nineteen human resource strategies currently being implemented, the research did not identify a strong association between any of these and job satisfaction or likelihood of leaving their job . In factResearchers also studied Burkina Faso\u2019s regionalized health personnel recruitment policy and questioned the sustainability of the policy given the absence of incentives . The resThe task-shifting and retention and recruitment research conducted within the context of the AHSI-RES program has uncovered important areas of focus for refining current human resources for health strategies, and approaches to evaluate whether these are producing the intended results. They also raised important issues to consider. Retention and recruitment strategies may not have an effect if they are designed without considering the needs or preferences of health workers. Likewise, task-shifting is not necessarily a solution in itself, but must be complemented by proper training, support, supervision, monitoring and evaluation, and a clear understanding of scope of practice. This indicates a need for clear guidelines, regulation, and recognition of the work of task-shifted health workers.Adrijana Corluka and Ren\u00e9e Larocque serve as Senior Program Officers, Marc Cohen serves as Program Officer, and Esm\u00e9 Lanktree serves as Program Management Officer, with the Global Health Research Initiative. The Global Health Research Initiative supported the assembly and publication of this supplement. The views expressed in this introductory article are those of the authors alone and do not represent the views of the Global Health Research Initiative, the International Development Research Centre, the Canadian Institutes of Health Research, nor Foreign Affairs, Trade and Development Canada."} +{"text": "We propose a mathematical model for multichannel assessment of the trial-to-trial variability of auditory evoked brain responses in magnetoencephalography (MEG).et al., our approach is based on the maximum likelihood estimation and involves an approximation of the spatio-temporal covariance of the contaminating background noise by means of the Kronecker product of its spatial and temporal covariance matrices. Extending the work of de Munck et al., where the trial-to-trial variability of the responses was considered identical to all channels, we evaluate it for each individual channel.Following the work of de Munck Simulations with two equivalent current dipoles (ECDs) with different trial-to-trial variability, one seeded in each of the auditory cortices, were used to study the applicability of the proposed methodology on the sensor level and revealed spatial selectivity of the trial-to-trial estimates. In addition, we simulated a scenario with neighboring ECDs, to show limitations of the method. We also present an illustrative example of the application of this methodology to real MEG data taken from an auditory experimental paradigm, where we found hemispheric lateralization of the habituation effect to multiple stimulus presentation.The proposed algorithm is capable of reconstructing lateralization effects of the trial-to-trial variability of evoked responses, i.e. when an ECD of only one hemisphere habituates, whereas the activity of the other hemisphere is not subject to habituation. Hence, it may be a useful tool in paradigms that assume lateralization effects, like, e.g., those involving language processing. Measurements of evoked brain responses in humans are an established standard in magneto- (MEG) and electroencephalography (EEG). The measured quantity is either the magnetic field above the surface of the head in MEG or the ei-th channel, at the j-th time moment, in the k-th trial, can be described as In the framework of the SPN model, the signal i\u2009=\u20091,\u2026,I, j\u2009=\u20091,\u2026,J, k\u2009=\u20091,\u2026,K. Rij is the brain response to the stimulus, which is assumed to be constant from trial to trial, and \u03b5 is a Gaussian noise with zero expected value, whose amplitude is typically larger than the amplitude of the evoked brain response. Due to the assumption of the Gaussian, zero-mean nature of the noise, \u03b5 is supposed to vanish on averaging across a large number of Rk), is replaced by Therefore, the order of the matrix Additionally, the following assumption is made K is the number of trials, says that in the case of non-variability of the response, each of the components on the leading diagonal of \u03c8k) poses further difficulties, because the dimensionality of the estimation process increases, yet we hope it offers a more accurate reflection of reality. Since it is known that the brain response evoked by a characteristic sensory stimulus arises in a particular area of the cortex , thede sites .et al.; SNR: Signal-to-noise ratio; SPN: Signal-plus-noise [model].The authors declare that they have no competing interests.X and T, performed most of the analyses, and wrote the paper. PK derived the estimators of \u03c8 and R, performed some analyses, and contributed substantially to interpretation of data; he also critically revised the model and the manuscript. Both authors read and approved the final manuscript.CS proposed the model and the estimation algorithm, derived the estimators of 1Special Lab Non-invasive Brain Imaging, Leibniz Institute for Neurobiology, Brenneckestr. 6, 39118 Magdeburg, Germany. 2Team Normal and Abnormal Motor Control, ICM Brain and Spine Institute, Sorbonne University, Pierre-and-Marie-Curie University (Paris 6), INSERM UMR1127, CNRS UMR7225, H\u00f4pital Piti\u00e9 Salp\u00eatri\u00e8re, 47 bd de l\u2019H\u00f4pital, 75013 Paris, France. C. Sielu\u017bycki started working on this project while he was with the Special Lab Non-invasive Brain Imaging, Leibniz Institute for Neurobiology, Magdeburg. He is now with the Team Normal and Abnormal Motor Control, ICM Brain and Spine Institute, Sorbonne University, Pierre-and-Marie-Curie University (Paris 6), INSERM UMR1127, CNRS UMR7225, Paris. 3 Department of Biomedical Physics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw, ul. Ho\u017ba 69, 00-681 Warszawa, Poland. P. Kordowski is currently a trainee in the Special Lab Non-invasive Brain Imaging, Leibniz Institute for Neurobiology, Magdeburg."} +{"text": "I am honored to assume the responsibility of the Editor-in-Chief (EIC) of the Journal of Epidemiology. For the last 6 years, 2002-2007, the editorial committee has been successfully managed under the strong leadership of the former EIC, Dr. Yosikazu Nakamura. During his term as the EIC, the journal has achieved the fundamentals required to become an international scientific journal. The journal successfully obtained an impact factor in 2005 in the first attempt and scored 1.240 in 2006. The journal has been issued bimonthly since 2000 and distributes over 30 articles annually. Its successful management is reflected in the extraordinarily speedy responses after article submission: the first reviewers' comments reach authors within an average of 18 days since the receipt of an article.The journal will continue to serve the research community across a broad range of exciting developments in epidemiologic research. First, I ask all members of the Japan Epidemiological Association to continually contribute to the journal by submitting significant research experiments. However, this alone is insufficient to further establish our excellence as an international scientific journal. Although based in Japan, the journal welcomes contributions from all over the world, especially from Asian countries. The importance and significance of the work it publishes is reflected in its international subscriptions and readership outside Japan. In order to maintain this high standard, the editorial committee will continue to make every effort to improve the quality of the journal.My own field of study is cancer epidemiology. However, I can assure you that the journal is not about to become a journal of cancer. We shall publish the best of all epidemiologic researches.Tomotaka Sobue, MD, MPHEditor-in-ChiefThe Journal of EpidemiologyCancer Information Services and Surveillance DivisionCenter for Cancer Control and Information ServicesNational Cancer Center5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, JapanPhone: +81-3-3542-2511, ext: 3424Fax: +81-3-3546-0630E-mail: edit-je@cied2.res.ncc.go.jp"} +{"text": "Communicating our Discipline: Strategies for the Emerging Information Infrastructures.Annually, the National Institute of Standards and Technology (NIST), Department of Commerce (DOC), and the National Computer Security Center (NCSC), National Security Agency (NSA), co-sponsor the National Computer Security Conference. The conference, most recently in its 17th edition (NCSC17), is a major event on the computer security conference calendar, bringing attendees together with leaders in the field, who report on their research and share experiences. Reflecting the need to more fully appreciate and practically deal with the major technical and social waves of change that we are experiencing, the theme of this year\u2019s conference was A large, diverse national and international audience attended the conference, with approximately 2,000 representing government, industry, and academe. NCSC17 provided a forum for technology interchange among system developers, and an opportunity for computer users to exchange ideas and learn about the latest methodologies to apply current computer and information security technology. Many reported that among the most valuable aspects of the conference was the opportunity for contemporaries to network, share information and experiences, and gain new perspectives through the conference\u2019s many and varied activities.The Learning Track, explored a variety of issues concerning information technology (IT) security education, training, awareness and professional development. There were also two special sessions devoted to progress on international harmonization efforts and the Common Criteria project. This year, main conference tracks focused on research and development, architecture and standards, applications and integration, management and administration, and tutorials for those new to the computer security field. Each track provided eleven At another special session, chaired by NIST\u2019s Computer Systems Laboratory Associate Director for Computer Security, F. Lynn McNulty, NIST announced and explained a set of NIST and DOC positions on the availability and applicability of the Digital Signature Standard, highlighting that the standard can now be used without fear of copyright infringement concerns.Security, Privacy, and Protection Issues in the Emerging Information Infrastructure.The opening plenary session featured an address by Sally Katzen, Administrator of the Office of Management and Budget\u2019s (OMB\u2019s) Office of Information and Regulatory Affairs, the presentation of the Conference\u2019s annual System Security Award, and an address by the awardee, Donn B. Parker of SRI International. Ms. Katzen offered insights about the role of the national information infrastructure (NII) in the changing way government does business and delivers its services. Mr. Parker spoke to the need for the security community to broaden its perspective beyond the traditional emphasis on confidentiality, integrity, and availability. (See Sec. 3.) The closing plenary session offered a lively discussion among a panel of distinguished experts on the subject of Research and Development Track traditionally addresses technical R&D efforts, including security models and intrusion detection. As in past years, intrusion detection was a significant area of interest. In a session chaired by R. Bace, NSA, intrusion detection was examined from the perspectives of design methodologies, a model for pattern matching, and current and future applications of artificial intelligence. (See Sec. 4.)The Two sessions explicitly addressed another traditional area of interest in this track, access control . A panel session, chaired by H. Feinstein, SETA Corp., looked at the future of role-based access control (RBAC), in terms of structure, mechanisms, the environment in which they operate, and how RBAC differs from the traditional trusted system security model .In another session, chaired by D. Cooper, Unisys, one paper described a specific access control model for achieving separation of duties, and two other papers examined architectures for RBAC and a means of implementing RBAC in a trusted on-line, transaction processing environment.Another panel, chaired by R. Nelson, Information Systems Security, explored non-traditional strategies for using fuzzy security as a means for building flexibility and functionality into trusted systems from a risk management perspective.In related sessions, M. Schaefer, Arca Systems, Inc. (Arca), chaired a paper session which looked at models addressing the development of secure database systems, and B. Thuraisingham, MITRE Corp., chaired a panel which focused on the inference problem in these systems.Key Escrowing: Today and Tomorrow, chaired by M. Smid, NIST; and The Security Association Management Protocol (SAMP), chaired by Maj T. Hewitt, USAF6, NSA, the latter addressing security services for communications. Another session presented papers on security in networks and distributed systems, chaired by D. Schnackenberg, Boeing Defense & Space Group; and still another, chaired by S. Jajodia, George Mason University, offered papers on formal methods and modeling regarding secure systems.Other sessions in this track included panel sessions on: Highlights of the New Security Paradigms \u201894 Workshop. Topics included were: fuzzy patterns in data; a health information architecture; applying formal semantics in multilevel logic databases; and the relationships among communication, information security, and value.To share the learnings from other IT security forums, a panel chaired by E. Leighninger presented the Architecture and Standards Track, new to this year\u2019s conference, focused on a variety of architectures and standards that are evolving to deal with emerging technical environments in the federal (DoD and civilian) and private sectors.The The Development of Generally Accepted System Security Principles (GSSPs). GSSPs were among a set of recommendations made in a National Research Council Study Report, Computers at Risk, published in 1991. (See Sec. 2.4.) Discussed were the GSSPs that NIST is developing under the auspices of Information Systems Security Association (ISSA) in coordination with OMB and with technical assistance from NSA.One panel, chaired by M. Swanson, NIST, addressed In a paper session chaired by W. Jansen, NIST, attendees learned about three differing approaches to security standards, including a taxonomy for viewing and developing them, the use of graphical displays and symantic networks, and vulnerabilities in the use of random pronounceable password generators.Two other sessions presented varying perspectives on issues related to national and international security criteria and assurance. A panel session chaired by P. Toth, NIST, included results of two workshops on assurance, and a paper session chaired by G. Wagner, NSA, looked at the development history for related procurement guidance.A panel session co-chaired by E. Flahavin, NIST, and J. Sachs, Arca, examined new challenges for certification and accreditation (C&A) from a variety of government perspectives, especially in environments where system and product interconnectivity and interoperability are at issue. An international panel, chaired by K. Keus, German Information Security Agency (GISA), Germany, looked at product and system certification from the perspectives of representatives of certification bodies of the European Community. Three sessions focused on security architectures. A panel session chaired by W. T. Polk, NIST, discussed the evolving Department of Defense (DoD) Goal Security Architecture, which reflects requirements for the support of multiple security policies, distributed information processing, conductivity by common carriers, users with different security attributes, and resources with varying degrees of security protection. A paper session chaired by H. Weiss, SPARTA, Inc., and a panel session co-chaired by R. Schell, Novell, GSA, and B. Dwyer, Hewlett-Packard, DCE, focused on related concerns in networked environments, including prominent industry-sponsored security architectures currently under development.In a two-session minitrack, chaired by J. Sheldon, USA, DISA/CISS, panelists explored current applications and future directions of multilevel security (MLS) . Included was an overview of the NSA Multilevel Information System Security Initiative (MISSI). (See Sec. 6.)Applications and Integration Track. Of special interest at this year\u2019s conference were the Internet, the NII, and how to achieve security in these environments. One approach to Internet security is through the use of a \u201cfirewall,\u201d and the wide area network (WAN) or global area network (GAN)). An overflow panel session, chaired by J. Wack, NIST, discussed how firewalls work, security policies that can be implemented by means of firewalls, and comparisons of how different firewall configurations support restricted access. Similarly, Provisions to Improve Security on the Internet, chaired by J. David, examined what the Internet has done to promote network security, and what steps can be taken to quickly and easily reduce specific risks. Can Your Net Work Securely?, chaired by P. Neumann, SRI, examined issues related to the often occurring situation in which distributed systems need to rely on components whose trustworthiness cannot be assured.The means by which security technology is being applied and how security products are being evaluated and integrated into secure systems was the focus of the Operational Security Enhancements, chaired by D. Dodson, NIST, provided a set of papers which looked at ways to improve security in Unix and C2 DOS/Windows-based personal computers, as well as a hardware device for system/data integrity and malicious code protection. Providing multi-vendor interoperability among security-enhanced and traditional UNIX systems was covered in the Trusted Systems Interoperability Group panel, chaired by S. Wisseman, Arca, which looked at the TSIG\u2019s related efforts since 1989.Proven Detection Tools For Intrusion Prevention, chaired by M. Higgins, DISA/CISS, took the audience through detection scenarios and lessons learned from the operational implementation of tools.Complementing the intrusion detection presentations (see Sec. 2.1), Putting Trusted Products Together, chaired by B. Burnham, NSA, including ways to approach analysis partitioning and composition analysis. Acquiring MLS system solutions was further debated among the key players in a panel session chaired by J. Sachs, Arca. Paper session Security Implementations, chaired by J. Anderson, J. P. Anderson Co., described a variety of security implementations,including thoseinbattlefield, customer network, and academic computing environments. The latter describing a mechanism that was developed to connect dispersed computing resources to achieve distributed processing while not eliminating local control. Panel session, NSA Concurrent Systems Security Engineering Support, chaired by B. Hildreth, NSA, looked at NSA\u2019s Test & Evaluation Community Network, which must evolve the capability for simultaneously processing unclassified and classified data while supporting both cleared and uncleared users.Various aspects of system integration were further addressed in Views on Vulnerability, chaired by R. Wood, NSA, addressed computer system vulnerabilities by looking at: evaluating information in computerized alarm systems; a tool for C&A support in DoD automated information systems (AISs); and using a financial management approach for selecting risk management-based safeguards.Finally, Management and Administration Track concentrated on subjects in the management and administration of the security function and the information systems which they support.The Model Information Security Programs, chaired by R. Owen, Jr., Texas Office of the Attorney General, examined IT security programs from the state, federal, private, and academic sectors, highlighting their similarities and differences in areas such as requirements, security organization structure, security management process, and methods of security awareness.Security in organizations can be improved by learning from the experiences of and modeling programs after those that have robust security programs in place. To this end, an informative and lively session, Interdisciplinary Perspectives on INFOSEC. In this panel, a diverse group of academics and practitioners, presented their thoughts as to how the insights of other disciplines can benefit the practice of IT security. Perspectives included anthropology, military science, ethics, psychology, theology, organizational development, and adult learning theory.Continuing to explore the theme of his paper on social psychology and information security that won an Outstanding Paper Award at NCSC16, M. E. Kabay, National Computer Security Assn., chaired Medical Information Privacy: Current Legislative andStandards Activities, chaired by M. Schwartz, Summit Medical Systems, Inc., which examined the technical and human issues generated by the currently available technology and practices in the medical arena.Privacy continued to be a subject of strong interest in this track. A particularly lively discussion took place in Ethical Issues in the National Information Infrastructure, chaired by J. Williams, MITRE Corp., and Detecting and Deterring Computer Crime, chaired by J. Holleran, NSA. The former explored broad issues, such as equity vs risk, privacy vs accountability, privacy vs surveillance, and international ramifications. The latter paper session looked at intrusion threats, detection using application profiles, and computer crime deterrence. (See Sec. 4.) A third session, Computer Crime on the Internet, chaired by C. Axsmith, Esq., ManTech Strategies Associates, provided many views of these subjects as they apply to the Internet. A fourth session, Risks and Threats, chaired by D. Gambel, Northrup Grumman, was geared to better understanding the elements of security threats and improving the assessment of risks.Another major theme in this track concerned computer ethics and computer crime. Among the sessions that directly addressed these areas were: Current Issues & Trends in Trusted Product Evaluations, a panel chaired by K. Bruso, NSA. Emphasis was on significant accomplishments in the area of trusted product evaluations during the past year, with special attention focused on two NSA assessment and evaluation programs.The importance of process improvements was explored in Do You Have the Skills to be a Future INFOSEC Professional?, chaired by W. Maconachy, DISA/CISS, viewed, from the federal government, private sector, and academe perspectives, the types of skills and individual initiatives needed to keep pace with changing work environments and advancing technologies and management challenges.There is a growing appreciation of the role of the IT security professional in the operation of the AIS function and ensuring business continuity. The panel, Real Lessons, chaired by J. Campbell, NSA, presented the audience with the lessons learned from real-world experiences in implementing security programs. Papers in this session addressed: security awareness in the persuasion of managers; the importance of workable network memorandums of agreement; and independent validation and verification of AISs.Another session, Computers at Risk (CAR) Recommendations: Are They Still Valid?, chaired by H. Tipton, CISSP, Member of the CAR Committee, Member of the GSSP Committee, provided a panel discussion that combined recent historical perspective and the lessons of practical experience. In this session, former members of the CAR committee revisited their recommendations in view of today\u2019s information security environment.In an exciting conclusion to this track, Each year\u2019s conference features a tutorial track. This popular track provides newcomers to the field and others wishing to acquire or \u201crefresh\u201d basic security subject matter an opportunity to do so.Trust Concepts, presented by C. Abzug, Information Resources Management College; Trusted Networks, presented by R. K. Bauer, Arca; Trusted Databases, presented by G. Smith, Arca; and System Security Engineering, Certification, and Accreditation, presented by J. Sachs, Arca, which focused on engineering and assessment issues in integrating MLS solutions using trusted products. C. Abzug additionally presented Criteria Comparisons, which focused on the differences and similarities of the national and international criteria of Canada, the United States, and Europe, in terms of value to security engineering, and as foundations for the Common Criteria.As in the past, this year\u2019s conference offered a set of tutorials on trusted systems, covering such subjects as: \u2014Risk Management and Security in the Future. The former focused on the overall risk management process, and the latter, co-presented with J. Sachs, Arca, looked at IT security and its role with respect to enterprises, applications, and information infrastructures.Two tutorials were presented by LtCdr A. Liddie, Royal Navy, Information Resources Management CollegeUNIX Security, presented by E. Schultz, Arca, and Windows NT Security, presented by J. Williams, Arca. Another tutorial, Information System Security Officer\u2019s Challenges, presented by C. Bressinger, DoD Security Institute, focused on the ongoing protection and accreditation of operational AISs. A concluding panel in this track, IT Security Resources, chaired by K. Everhart, NIST, offered attendees an overview of major electronic and nonelectronic sources of information on IT security and a discussion of emerging software standards to disseminate and access security-relevant information resources.Two other tutorials addressed security in specific software environments \u2014 Standards and Architecture Track, two special sessions were held this year related to international harmonization and the Common Criteria (CC). The CC project refers to the work performed by the U.S. and the European Communities (EC) to develop a common basis for evaluating the ability of products and systems to protect confidentiality of data and provide other security controls. Such evaluations are expected to reduce costs to users and vendors.The conference continued to reflect community interest evaluation criteria. In addition to the panel and paper sessions discussed in the International Harmonization, the Common Criteria\u2014Progress & Status, chaired by E. Troy, NIST, representatives from the European Commission , Canada, and the United States discussed the CC project, schedules, documents used as input, and the public review process. An overview of the draft Common Criteria document was also presented.In the first special session, Security Requirements for Distributed Systems, chaired by R. Dobry, NSA, panelists from NIST, NSA, the University of Maryland, and the Institute of Defense Analysis identified requirements for providing security for distributed systems and how they saw their efforts relating to the Common Criteria.In the second special session, The Learning Track. Meaningful security education, training, and awareness for all, and the availability of staff who can ensure that appropriate controls are in place, are part of an overall resource management strategy. The track was framed against the backdrop of an environment that is being shaped by both the emergence of the NII and increasing pressures on all to be more productive. There is also a renewed appreciation by public and private sector organizations about the need to cost-effectively protect information systems resources. The sessions focused on several efforts throughout the IT security community relating to learning initiatives and the professional development of security practitioners. The NIST-sponsored Federal Information Systems Security Educators\u2019 Association (FISSEA) and the National Security Telecommunications and Information Systems Security Committee (NSTISSC)-sponsored Information Systems Security Education, Training, and Awareness (ETA) Working Group coordinated the track.Another feature of this year\u2019s conference was Training Challenges of the 90s, chaired by J. Pohly, CISS/FISSEA Chair, addressed the security demands that the NII will place on the workforce and the security professional, identified the challenges of complying with training mandates, and outlined proposed solutions.To introduce the track, Proposed New NIST Training Standards, chaired by D. de Zafra, Public Health Service, a draft developed by FISSEA that is proposed to replace the NIST training guideline, NIST Special Publication 500-172, was discussed.Training standards are seen as one element of the solution equation. In Computer Security Resources that Work, chaired by B. Cuffie, Social Security Administration; Tools and Methodologies for Delivering Training, chaired by J. Jelen, Public Health Service; and Demonstrations on Computer Security Training Tools, chaired by A. Stramella, National Cryptologic School. The latter focused on computer-based training packages, videos, and interactive learning tools.A number of sessions in the track looked at the tools, resources, and methodologies for developing and delivering IT security training, and reported on related experiences. These sessions included: Effective Marketing of the Computer Security Program to Management, chaired by J. Hash, Social Security Administration, and Training Events on a Shoestring Budget, chaired by S. Pitcher, Department of Commerce, as panelists shared their real-world experiences.Two issues with which those responsible for IT security programs and security education, training, and awareness must continually deal are garnering management support, and relatedly, competing for budget dollars to implement programs. These issues were addressed in Information Systems Professionalism \u2014 Professional Development and Certification, co-chaired by R. Koenig and H. Tipton, International Information Systems Security Certification Consortium (ISC)2, explored the current status and future directions of several initiatives underway to professionalize the community and certify the computer security professional.Professionalization and certification are increasingly recognized as integral to how the profession grows, nurtures, attracts, and retains IT security practitioners. Adult Learning and Information Systems Security Training, presented by E. Martin, Organization and Education Consultant. This session continued with the theme that IT security can benefit from the learnings of other disciplines. The session reviewed recent developmentsin methodology that offer more effective ways of teaching adults to use technical skills that also require individual judgement. It drew on the research and experiences of employer-sponsored training to examine lessons learned about methodologies in use, the basic concepts of adult learning, and the ways these principles can be applied to information systems security training. Concepts were demonstrated by means of experiential exercises.A particularly interesting session in this track was Security, Privacy, and Protection Issues in Emerging Information Infrastructures. The panel was co-chaired by Professor Anthony Oettinger, Chairman, Program on Information Resources Policy, Harvard University, and Dr. Brian Kahin, Director, Information Infrastructure Project, Science, Technology and Public Policy Program, Harvard University. Other panelists were Robert Lucky, Vice President Applied Research, Bellcore, and Robert Wilson, MCI.The closing plenary featured a distinguished panel addressing This interesting panel included lively exchanges among panel members and between the audience and the panel. Each panel member started with a brief statement of issues and perspectives. Overlapping topics included technology advances, pending and possible legislation, intellectual property concerns, major stakeholders, market restructuring, the convergence and integration of media and delivery systems, market share and other business concerns, protection of individual privacy and corporate/organizational proprietary information, other security and protection concerns, standards of due care, and global rather than national scope of the problems ). One interesting comment came from audience member M. Kabay who said that not until insurance companies punished/rewarded those that avoided/embraced standards of appropriate care to protect their information systems, would significant progress be made. He challenged the community to exert the needed pressures to make that happen.A particularly satisfying event of each year\u2019s conference is the recognition of an individual who has contributed significantly to the computer security community over an extended period of time. The recipient of this year\u2019s Systems Security Award was Donn B. Parker, Senior Management Consultant, SRI, International. Mr. Parker has conducted extensive research on the human and technical factors involving cases, causes, and the prevention of computer crime, and has promoted a philosophy that security must be treated as a \u201cpeople\u201d problem, in addition to a technical problem. His research and five books have addressed computer crime, ethics, and information security management. He has contributed to many professional organizations in a variety of capacities. An international lecturer and management consultant, he has served leading businesses, the U.S. Congress, state legislatures, and government agencies. He also created SRI\u2019s International Information Integrity Institute (14), which provides services to 60 of the world\u2019s largest corporations.Mr. Parker joins a distinguished list of previous Systems Security Award winners which includes Stephen Walker, Dr. Willis Ware, James P. Anderson, Dr. Roger Schell, Dr. Walter Tuchman, and Robert Courtney.Can Computer Crime Be Deterred?. This is one of the few papers that has addressed the critical questions of whether computer crimes can be deterred and, if so, by what means. The author points out that we tend to emphasize the computer aspects much more than the criminal aspects in the prevention of computer crime. While deterrence is difficult to achieve, information security programs have essentially neglected attempts to make it work. The author reviews the research findings from criminological and legal studies of deterrence and applies these findings to computer crime prevention. Legislative, law enforcement, and organizational changes need to be made to effectively deter computer criminals. The author makes the case for changing the perceptions of employees and outsiders regarding the risks of getting caught in computer crime, as well as the perceived payoffs from such activities.Two outstanding paper awards were presented at this year\u2019s conference. One went to Sandford Sherizen, Ph.D., Data Security Systems, Inc., for his paper Artificial Intelligence and Intrusion Detection: Current and Future Directions. Identification of attempted or ongoing attacks on computer systems and networks, or intrusion detection, is a growing concern for users and administrators of these systems, who rely on their secure operations. This concern increases with each new reported Internet attack. Previous approaches \u201cby hand\u201d to intrusion detection systems (IDSs) made it difficult to create robust, real-time systems. The author notes artificial intelligence (AI) techniques can be effectively applied to the problem, and surveys the methods by which this has been done. The difficulty and computational intensity involved with the activities of data reduction and behavior classification are described. Significantly, the author demonstrates how the use of the technique of feature selection can reduce computational overhead and improve classification of network connections.The other outstanding paper award went to J. Frank, University of California, Davis for his paper Testing Intrusion Detection Systems: Design Methodologies and Results from an Early Prototype, and S. Kumar, Purdue University, (co-author and advisor E. Spafford) for A Pattern Matching Model for Misuse Intrusion Detection.Each year, the conference committee invites teachers in IT security-related disciplines to submit papers written by students in a degree program who have not been previously published. This year the conference program committee recognized two excellent student papers. The awards, both for papers in the area of intrusion detection, went to N. Puketza, University of California, Davis, for As in past years, the conference held a joint awards ceremony in which NIST and NCSC honored the vendors who had successfully developed products meeting the standards of their respective organizations. In the case of NIST, its Computer Security Division provides validation services for vendors to use in testing devices for conformance to security standards defined in three Federal Information Processing Standards (FIPS): Data Encryption Standard (DES), Computer Data Authentication, and Key Management Using ANSI X9.17. This year, many of the NIST awards were for the recently permitted software implementations of DES. In the case of NCSC, vendors are recognized who contribute to the availability of trusted products and who thereby expand the range of solutions customers can use to secure their data. The products are placed on the Evaluated Products List (EPL) following a successful evaluation against the Trusted Computer Systems Evaluation Criteria and its interpretations. A number of other special sessions and demonstrations were available to attendees. These are listed in the following sections.Dr. Corey Schou, of Idaho State University (ISU), has developed an electronic group decision support system that has been effectively applied to a wide range of IT security questions, issues, and challenges. This has been demonstrated through a series of DACUM (Design-a-Curriculum) workshops at ISU, the results of which are contributing to the development of security awareness training materials; IT security curricula; a proposal to revise the NIST Training Guidelines (SP 500-172) with a more rigorous conceptual model for security training; a unified body of knowledge for security practitioners; and knowledge, skills and abilities (KSAs) and plans of instruction for various security-related job categories. The consensus among those who have taken part in the DACUMs, is that the technology can be effectively applied to a wider range of IT security questions, issues, and challenges beyond the DACUM arena.This year, a portable, ten-station version of the system offered attendees the opportunity to participate in demonstrations and to \u201ctest drive\u201d the system. They were able to view the results of the DACUM workshops and a large archive of security information developed at ISU to support the DACUMs. Two groups, one from the GSSP Committee and the other revising the NIST Training Guidelines, each reserved a session on the system to collaboratively \u201cbrainstorm\u201d about elements of documents being developed. The former used the system to define the meaning of so-called \u201cpervasive principles,\u201d and the latter for learning objectives associated with AIS functional areas throughout the system life cycle.The TSIG offers an open forum for developers of secure networking systems and those who have a shared vision of making open trusted systems a reality. The MISSI is evolving a series of products which, when combined, provide security services for a variety of MLS environments. These vendor demonstrations showed how many different MLS hardware devices and applications are used in stand-alone and integrated, real-world environments.The Information Technology Security Evaluation Facilities (ITSEF) in Europe and the European certification bodies reported on the system and security product evaluations being performed under its program, and demonstrated the product evaluation methodology.CISS, which is jointly-staffed by DISA and NSA, presented displays and demonstrations to showcase services and products that directly support DoD, including demonstrations by the Automated Systems Security Incident Support Team (ASSIST).Presented were overviews of Air Force system security initiatives, including demonstrations on intrusion detection and risk management.This workshop consisted of several short presentations and discussion periods, including progress reports on development projects; experiences; auditing, legal, privacy, and network security issues; intrusion scenarios; new detection techniques; incident response; and intrusion detection systems requirements.Other activities of interest at the conference included NIST and NSA awareness and information booths where a variety of technical and other publications from each organization were available, including NIST Computer Systems Laboratory (CSL) Bulletins and NSA\u2019s Rainbow Series; demonstrations of NSA\u2019s Dockmaster and NIST\u2019s Computer Security Resources Clearinghouse, each of which offers a wide variety of IT security information through dial-in and Internet access; demonstration of a computer-aided instruction course that was developed by NSA to provide basic-level security information; a combined book exhibit representing a selection of leading publishing firms and the latest selections in computer security, presented by Association Book Exhibit; a booth at which the professional organization Information Systems Security Association (ISSA) presented information and recent newsletters, resource guides, and technical publications; and birds-of-a-feather (BOF) rooms which were used by groups to address self-defined areas of interest.Next year\u2019s conference, the 18th in the series, will be named the National Information System Security Conference. NISSC18 will be held October 10\u201313, 1995 at the Baltimore Convention Center. For further information, contact the NIST Conference Office, (301)975-2775.Single copies of the 742-page NCSC17 conference proceedings are available upon request. Please contact NIST CSL Publications at (301)975-2821.We are considering putting future conference proceedings, plus additional security-related information, on a CDROM, along with appropriate retrieval capability. Our objective is to keep the price at a minimum, but sufficient to cover expenses. For further information, please contact: (301)975-3359."} +{"text": "SICOT-J has now been launched. It is backed by an internationally renowned Editorial Board and we seek quality publications.Over the last few years, Orthopaedics has been inundated with Open Access journals, the continued quality and survivorship of which is uncertain. SICOT, the only international society of Orthopaedics and Traumatology has therefore decided to participate and take a lead in this rapidly evolving field. SICOT-J is an Open Access journal which publishes original clinical, basic and translational research in orthopaedic surgery and traumatology. SICOT-J will accept, subject to peer review, review articles, original articles, research articles, surgical techniques, case reports, congress proceedings, editorials and letters to the Editor.SICOT-J is published under Open Access conditions, all the articles will also be published quickly by the Open Access Repository (OAR) of publicly accessible scientific articles, in Biomedical and Life Science journals and by PubMed Central which is hosted by the US National Institute of health\u2019s National Library of Medicine (NIH/NLM).As SICOT-J by visiting the website of the journal at www.sicot-j.org. We hope that many of you will support the Journal as it is with your quality articles that the journal\u2019s impact in the field of orthopaedics and traumatology will quickly flourish.We invite you to submit your articles to The Editors-in-chiefJacques CatonHatem Said"} +{"text": "This article adds Yann Le Franc as a co-author of the technology report article \u201cSoftware and Hardware Infrastructure for Research in Electrophysiology,\u201d describes his individual contribution to the article, and presents changes in two paragraphs in Section 2.4, where an additional reference is also provided. Moreover, the members from the OEN group who work and cooperate on building OEN are personally acknowledged.Co-author: Yann Le FrancAffiliation: e-Science Data Factory S.A.S.U., Paris, France; Ludwig-Maximilians-Universit\u00e4t M\u00fcnchen, Planegg-Martinsried, Germany; University of Antwerp, Antwerpen, BelgiumAuthors' ContributionsFigures 3,4 and 8 and to the work on the Ontology for describing Experimental Neurophysiology (OEN).Yann Le Franc contributed to the Section 2.4Old version:The group follows the best practices for creating ontologies, for example, it cooperates with community of researchers who design and create ontologies, uses existing data formats and repositories , and reuses existing resources . For the general description of experimental neurophysiology, the terms from ontologies NEMO and OBI are relevant. However, the set of the domain terms is still not complete in these ontologies and OEN will be finally an extension of OBI .New version:The group follows the best practices for creating ontologies, for example, it cooperates with community of researchers who design and create ontologies, uses existing data formats and repositories , and reuses existing resources . For the general description of experimental neurophysiology, the terms from ontologies NEMO and OBI are relevant. However, the set of the domain terms needed to describe the information stored in the EEG/ERP Portal is not yet complete in these ontologies. OEN aims at defining these missing terms and at term, should be used to propose an extension of OBI's neurophysiology model .Old version:https://github.com/G-Node/OEN.Terminologies within OEN have been primarily developed in the odML format. Subsequently, an OWL file has been constructed aided by Ontofox (Xiang et al., New version:https://github.com/G-Node/OEN.The OEN device branch development is based on the odML terminology (Grewe et al., The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "CDC and the Rollins School of Public Health at Emory University will cosponsor the course, Introduction to Public Health Surveillance, May 20\u201324, 2013, at Emory University in Atlanta, Georgia. The course is designed for state and local public health professionals.The course will provide theoretical and practical knowledge to design, implement, and evaluate effective public health surveillance programs. Topics scheduled for presentation include an overview and history of surveillance systems; planning considerations; sources and collection of data; analysis, interpretation, and communication of data; surveillance systems technology; ethics and legalities; state and local concerns; and future considerations. Tuition is charged.http://www.sph.emory.edu/epicourses); by e-mail ; by mail ; by telephone (404-727-3485); or by fax (404-727-4590).Additional information and applications are available online ("} +{"text": "Background: In recent years, electronic cigarettes (e-cigarettes) have generated considerable interest and debate on the implications for tobacco control and public health. Although the rapid growth of e-cigarettes is global, at present, little is known about awareness and use. This paper presents self-reported awareness, trial and current use of e-cigarettes in 10 countries surveyed between 2009 and 2013; for six of these countries, we present the first data on e-cigarettes from probability samples of adult smokers. Methods: A cross-sectional analysis of probability samples of adult (\u2265 18 years) current and former smokers participating in the International Tobacco Control (ITC) surveys from 10 countries. Surveys were administered either via phone, face-to-face interviews, or the web. Survey questions included sociodemographic and smoking-related variables, and questions about e-cigarette awareness, trial and current use. Results: There was considerable cross-country variation by year of data collection and for awareness of e-cigarettes (Netherlands (2013: 88%), Republic of Korea (2010: 79%), United States (2010: 73%), Australia (2013: 66%), Malaysia (2011: 62%), United Kingdom (2010: 54%), Canada (2010: 40%), Brazil (2013: 35%), Mexico (2012: 34%), and China (2009: 31%)), in self-reports of ever having tried e-cigarettes , Malaysia (19%), Netherlands (18%), United States (15%), Republic of Korea (11%), United Kingdom (10%), Mexico (4%), Canada (4%), Brazil (3%), and China (2%)), and in current use , Republic of Korea (7%), Australia (7%), United States (6%), United Kingdom (4%), Netherlands (3%), Canada (1%), and China (0.05%)). Conclusions: The cross-country variability in awareness, trial, and current use of e-cigarettes is likely due to a confluence of country-specific market factors, tobacco control policies and regulations , and the survey timing along the trajectory of e-cigarette awareness and trial/use in each country. These ITC results constitute an important snapshot of an early stage of what appears to be a rapid progression of global e-cigarette use. The availability of less toxic forms of nicotine delivery products may represent a new paradigm for tobacco control by offering smokers an opportunity to obtain nicotine in ways that do not require inhaling tobacco smoke ,2,3. Theet al. presented a qualitative overview of the ANDS literature about awareness and use, and highlighted gaps in knowledge [A recent systematic review by Pepper nowledge . The autnowledge , evidencnowledge ,18,19,20nowledge ,25,26,27nowledge ,29,30. Tnowledge ,32,33,34et al. [Only one study to date has presented an international comparison of e-cigarette awareness and use. Adkison et al. presenteet al.\u2019s analysis of four high-income countries [The current study is the first of its kind to compare levels of e-cigarette awareness, trial, and current use among national representative samples from multiple countries with different economic levels and with tobacco control environments. The current study is an extension of Adkison stralia) : the AusA cross-sectional analysis was conducted from 10 countries participating in the International Tobacco Control (ITC) Surveys: The Netherlands, Republic of Korea, United States, Malaysia, United Kingdom, Canada, Brazil, Mexico, China, and Australia. The latest cohort dataset available in each country, conducted between 2009 and 2013, was included. Respondents were adult (\u2265age 18) current or former smokers. Current smokers were those who reported having smoked at least 100 cigarettes in their lifetime and who had smoked at least one cigarette in the past 30 days; former smokers were those who were smoking at the time of study initiation but had quit smoking at some point over the course of study follow-up. Recent quitters had stopped smoking in the last 6 months or less.Methodological details for each country are available via the ITC Project website . In brieversus Not at all), The Netherlands: How often do you currently use an electronic cigarette? , and China: Are you currently using an electronic cigarette at least weekly? (Responses: Yes or No). For the countries that had multiple choices for current use, the variable was dichotomized into \u2018yes\u2019 or \u2018no\u2019 responses, where \u2018yes\u2019 corresponds to any form of use and \u2018no\u2019 corresponds to not being a current consumer (stopped or never used it).Questions about e-cigarettes included: (1) Have you ever heard of electronic cigarettes or e-cigarettes? (Responses: Yes or No); (2) Have you ever tried an e-cigarette? (Yes or No); and (3) Do you currently use e-cigarettes? The latter question about current use was not consistent between all countries. All responses were however dichotomized. Specifically, the question and responses for the Republic of Korea, US, UK, Canada, Australia, and Malaysia included: How often, if at all, do you currently use an electronic cigarette? . All other results were based on analyses on weighted data to allow for prevalence estimates. The weights of each country were constructed individually by means of its sampling plan. For example, if simple random sampling was used, then the weights were inflated step by step to the household level then to the city/province level, then finally to the country representative level. If two-stage stratified sampling was used, then the weights were first inflated to the household level then to the cluster level, then to the strata level. These weights may not be country representative, so all inflated weights were then rescaled to the country sample size. The rescaled weights for each country were pooled in the analytic model to allow for cross-country comparisons. For details of the weight construction process for each country, please refer to the ITC Technical Reports . Rates of e-cigarette awareness, trial, and current use were computed for smokers and former smokers. These rates were further broken down to present the proportion of current smokers and recent quitters (those that had quit smoking in the last 6 months) that were aware of, tried or currently used e-cigarettes. The analyses were conducted with SAS 9.2. Participant characteristics and survey details are presented in This study of 10 countries provides a snapshot of the early stage of what appears to be a rapid progression in the global use of alternative nicotine delivery systems, such as e-cigarettes. In 6 countries\u2014Netherlands, Brazil, Mexico, China, Republic of Korea, and Malaysia\u2014we report the first data on e-cigarettes from probability samples of adults. The findings demonstrate considerable cross-country variation in both awareness and use of e-cigarettes. Some of the variability in awareness, trial, and use of e-cigarettes appears to be related to a combination of between-country differences in when the surveys were conducted , the regulatory status of e-cigarettes , and levels of enforcement .vs. low-/middle-income. The relatively high rates in Malaysia run contrary to notions that e-cigarettes would be a less viable product in non-high income countries.Interestingly, some of the cross-country differences do not seem readily explained by standard categories of countries such as high- It may well be the case that although some of the between-country differences may be at least partially explained by legal restrictions, as discussed above, that the variability between countries may be due to conditions specific to those countries such as the presence or absence of entrepreneurs and companies to bring a product that was until very recently, almost unknown. The uncertainty in explaining these differences across countries points to the need for case studies of the reasons for the ascendency of e-cigarettes in some countries and the relative absence of such products in others. In China, there was a very low rate of e-cigarette use. Although this may be surprising given that e-cigarettes were invented in China in 2003, and more than 90% of the e-cigarettes worldwide are produced in China, it should also be noted that China is the home of the world\u2019s largest cigarette company, China National Tobacco Company (CNTC), which along with the State Tobacco Monopoly Administration, would have a strong interest in not supporting the emergence of e-cigarettes in China\u2014a non-CNTC product. However, the atmosphere may be changing for e-cigarettes in China. A recent news report states that the CNTC is in the process of launching its own line of e-cigarettes .These data confirm and extend previous knowledge of awareness, trial, and use in high-income countries (HICs). In particular they are consistent with data from large national surveys ,23,28 prThere are several limitations in this study. First, the timing of the surveys differed across countries, and so it is, as discussed above, difficult to make confident judgments about the reasons for differences across countries. Thus, the lower rates of e-cigarette awareness and use reported in some countries may be an artifact of when the survey was conducted since e-cigarette use has increased with time in all countries where there exist longitudinal data. Additionally, recent unpublished ITC data from Australia, Canada, UK, and US reveal a rapid increase in reported awareness and use of e-cigarettes between 2010 and 2013 . AnotherIn summary, this report provides a brief overview of e-cigarette awareness and use across 10 different countries with diverse economies and tobacco control histories. Additional research is needed to understand the patterns of global ANDS use in greater detail, particularly on impact of e-cigarettes on dual use, quitting, and relapse among smokers, and initiation of smoking among youth. There is a need for longitudinal studies to focus on the impact of ANDS, and address the critical question whether e-cigarettes represent a positive, negative, or mixed phenomenon for tobacco control and for public health. This will highly depend on studies that are able to measure and understand the interplay between ANDS and combustible tobacco products. The impact of ANDS marketing on the uptake of smoking by young people, along with the impact of e-cigarettes on smokers is necessary to make evidence-based judgments about the net burden of ANDS on the tobacco pandemic, within and across countries."} +{"text": "A suite of resources provides implementation guidance for mHealth initiatives, particularly in less developed countries. The suite includes an eLearning course, online guide, evidence database, and a High-Impact Practices brief, along with the mHealth Working Group and website. A suite of resources provides implementation guidance for mHealth initiatives, particularly in less developed countries. The suite includes an eLearning course, online guide, evidence database, and a High-Impact Practices brief, along with the mHealth Working Group and website. We believe this is indicative of the high level of interest in using mobile technology to strengthen health service delivery and to expand the reach of health promotion activities in countries around the world.We note with interest that the article \u201cWe would, therefore, like to introduce your readers to a suite of mHealth resources developed by the U.S. Agency for International Development (USAID) and its partner, the Knowledge for Health Project, with input from FHI 360 and the mHealth Working Group. The resources are designed to strengthen mHealth capacity of health program implementers and managers, particularly in less developed countries.www.globalhealthlearning.org/course/mhealth-basics-introduction-mobile-technology-health). This self-directed course,\u00a0mHealth Basics: Introduction to Mobile Technology for Health, explains the potential uses, benefits, and limitations of mHealth. It also highlights some existing applications, draws some preliminary conclusions from the evidence, and shares recommended best practices for mHealth program planning, design, monitoring and evaluation, scale up, and sustainability. It takes about 3\u2005hours to complete the course.First, we have developed a new online eLearning course as part of USAID's Global Health eLearning Center is designed to bring together published reports and articles on mHealth effectiveness, cost-effectiveness, and program efficiency. This collection of resources makes it easier for researchers, program managers, funders, and other key decision-makers to quickly get up to speed on the current state-of-the-art. It includes peer-reviewed and grey literature from high-, middle-, and low-resource settings. Materials are classified using a new mHealth Evidence Taxonomy, developed in coordination with the World Health Organization, and are easily filtered and searched to facilitate the identification of evidence-based, high-impact mHealth practices.The\u00a0mHealth: Mobile Technology to Strengthen Family Planning Programs, was recently published as part of USAID's High-Impact Practices in Family Planning series, which identifies mHealth as an emerging practice. This brief highlights evidence in mHealth and family planning programs to date and contains a synthesis of lessons learned for implementation of mHealth programs. The brief can be accessed at: www.fphighimpactpractices.org/resources/mhealth-mobile-technology-strengthen-family-planning-programsAn 8-page brief, mHealth Working Group and website . Started in 2009 with 20 members based in Washington, DC, the mHealth Working Group has grown into an international community of over 1,500 members representing more than 450 organizations in 70 countries. The group's mission is to frame mobile technology within a larger global health strategy and to build capacity, encourage collaboration, and share knowledge. Monthly meetings are held in Washington, DC, with invited speakers. Virtual participation in the meetings is also an option. The group's website contains presentations from the monthly meetings along with other resources.In addition to these 4 tools, another relevant resource is the Global Health: Science and Practice.We look forward to seeing more new research related to the roll out and scale up of mobile health applications in"} +{"text": "The \u201cVaccine and Drug Ontology Studies\u201d (VDOS) international workshop series focuses on vaccine- and drug-related ontology modeling and applications. Drugs and vaccines have been critical to prevent and treat human and animal diseases. Work in both (drugs and vaccines) areas is closely related - from preclinical research and development to manufacturing, clinical trials, government approval and regulation, and post-licensure usage surveillance and monitoring. Over the last decade, tremendous efforts have been made in the biomedical ontology community to ontologically represent various areas associated with vaccines and drugs \u2013 extending existing clinical terminology systems such as SNOMED, RxNorm, NDF-RT, and MedDRA, developing new models such as the Vaccine Ontology (VO) and Ontology of Adverse Events (OAE), vernacular medical terminologies such as the Consumer Health Vocabulary (CHV). The VDOS workshop series provides a platform for discussing innovative solutions as well as the challenges in the development and applications of biomedical ontologies for representing and analyzing drugs and vaccines, their administration, host immune responses, adverse events, and other related topics. The five full-length papers included in this 2014 thematic issue focus on two main themes: (i) General vaccine/drug-related ontology development and exploration, and (ii) Interaction and network-related ontology studies. Drugs and vaccines have been critical to prevent and treat human and animal diseases. Work in both (drugs and vaccines) areas is closely related - from preclinical research and development to manufacturing, clinical trials, government approval and regulation, and post-licensure usage surveillance and monitoring. Ontologies can serve important roles in managing, normalizing, sharing, and leveraging drug and vaccine relevant data. The 2014 \u201cVaccine and Drug Ontology Studies\u201d workshop (VDOS 2014) was an international forum for researchers to identify, propose, and discuss solutions for important research problems in ontology representation and analysis of vaccine and drug development, administration, function mechanisms, safety, and education.VDOS 2014 was held on October 7, 2014, in Houston, Texas. This workshop was part of the Fifth International Conference on Biomedical Ontology (ICBO 2014). The workshop attracted interests from many international attendees, including paper presenters, academic and government scientists, postdoctoral fellows, and graduate students. After a rigorous peer review process , five full-length papers were accepted for proceeding paper publications and oral presentations in the workshop. After additional reviewing by the independent reviewers, workshop co-organizers, and the journal editors, the selected five full-length papers were extended and accepted for publication in the thematic track of the Journal of Biomedical Semantics (JBMS).rd in this series. The first workshop of the series was organized as the \u201cVaccine and Drug Ontology in the Study of Mechanism and Effect\u201d workshop (VDOSME 2012) [The VDOS-2014 workshop is the 3The five papers selected for this thematic issue are extended versions of the original full-length papers presented at the VDOS 2014. This year the workshop focuses on two main themes: (i) General vaccine/drug-related ontology development and exploration, and (ii) Interaction and network-related ontology studies.In the area of general vaccine/drug-related ontology development and exploration, Winnenburg and Bodenreider introduced their approach on identifying common class-level signals for every individual drug in a class . They crIn the area of interaction and network-related ontology studies, Peters et al. investigated the drug \u2013drug interaction (DDI) information in two publically available drug resources, NDF-RT and DrugBank . Their sIn the 2014 VDOS workshop, the five full-length papers described above were orally presented. In addition, the program also included a keynote speech and a podium abstract presentation. The keynote speech was given by Dr. Khalid F. Almoosa, Associate Professor of Medicine at the Department of Internal Medicine, UTHealth Medical School, Houston, TX. Dr. Almoosa also serves as the Vice-Chair for Healthcare Quality and the Director of Interprofessional Collaboration at UTHealth. In his keynote, Dr. Almoosa discussed important issues in healthcare quality and safety and the roles of vaccine and drug safety in healthcare quality management. For the podium abstract presentation, Dr. Yongqun He provided updates on the development of the Ontology of Adverse Events (OAE) and its applications. An in-depth discussion of the OAE is available in the recently published article in Journal of Biomedical Semantics .The discussion session of the workshop focused on the topic of \u201cDrug and vaccine ontology informatics\u201d, we first discussed the similarities and differences between the adverse event ontologies in the Open Biological and Biomedical Ontologies (OBO) domain and non-OBO style ontologies . The necessity and possible approaches on how to bridge OBO and non-OBO ontologies were explored. Furthermore, the participants of the workshop discussed ontology-based clinical and experimental data processing and analysis, including ontology-supported literature mining of biomedical and electronic health records. We all agreed to continue the workshop in 2015 which would be held in conjunction with the ICBO-2015 conference at Lisbon, Portugal, in the end of July 2015.Overall, the VDOS 2014 workshop was well received with positive feedbacks. The VDOS workshop series have served and will continue to serve a platform for biomedical and clinical researchers to discuss ontology-relevant issues and progress in drug and vaccine research and applications."} +{"text": "There is an error in the Funding section of the article. The correct Funding statement is:\u201cThis project was supported by the National Science, Technology and Innovation Plan (NSTIP), Strategic technologies program, grant number (11-BIO1974-02), the Kingdom of Saudi Arabia. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "Designed for working professionals who are unable to leave their jobs, the Career Master of Public Health (CMPH) program at the Rollins School of Public Health, Emory University, is a distance-learning program. Each semester, students are required to attend two long weekends on the Emory University campus in Atlanta, Georgia. All additional coursework is taught using the Internet and distance-learning techniques.The CMPH program offers three majors: Applied Epidemiology, Applied Public Health Informatics, and Prevention Science. In addition to coursework in their major, students in the CMPH program take six required core courses and additional coursework that includes competencies in public health informatics, evaluation, and surveillance. All CMPH students also complete a practicum and culminating experience that are relevant to their areas of specialization.cmph@sph.emory.edu or visit online at http://www.sph.emory.edu/cmph.Program faculty members represent both academia and public health practice and include persons who work at Emory, CDC, and local public health departments. Completion of coursework for the CMPH degree takes 7 semesters for the full-time student and is fully accredited by the Council on Education for Public Health and the Southern Association of Colleges and Schools. Prospective students should apply by May 1, 2013, to guarantee consideration for the fall 2013 semester. Additonal information is available via e-mail at"} +{"text": "The names of the seventh and 12th authors were spelled incorrectly. The correct names of the seventh and 12th authors are Ju-Yeh Yang and Su-Ying Wen, respectively.In addition, multiple funding organizations were incorrectly given, and multiple funding organizations and grants were incorrectly omitted from the Funding Statement. The Funding Statement should read: \"This study was supported by research grants to Dr. Mei-Ju Ko, from the National Science Council of Taiwan (NSC 102-2314-B-532-002), Department of Health, Taipei City Government , and Taipei City Hospital, Ren-Ai branch . The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "Background: The spread of drug-resistant tuberculosis (TB) is one of the major public health problems through the world. Surveillance of anti-TB drug resistance is essential for monitoring of TB control strategies. The occurrence of drug resistance, particularly multi-drug resistance Mycobacterium tuberculosis (MDR), defined as resistance to at least rifampicin (RIF) and isoniazid (INH), has become a significant public health dilemma. The status of drug-resistance TB in Iran, one of the eastern Mediterranean countries locating between Azerbaijan and Armenia and high-TB burden countries (such as Afghanistan and Pakistan) has been reported inconsistently. Therefore, the aim of this study was to summarize reports of first-line anti-tubercular drug resistance in M. tuberculosis in Iran.Material and Methods: We systematically reviewed published studies on drug-resistant M. tuberculosis in Iran. The search terms were \u201cMycobacterium tuberculosis susceptibility\u201d or \u201cMycobacterium tuberculosis resistant\u201d and Iran.Results: Fifty-two eligible articles, published during 1998\u20132014, were included in this review. Most of the studies were conducted in Tehran. The most common used laboratory method for detecting M. tuberculosis drug resistant was Agar proportion. The highest resistance to first-line drugs was seen in Tehran, the capital city of Iran. The average prevalence of isoniazid (INH), rifampin (RIF), streptomycin (SM), and ethambotol (EMB) resistance via Agar proportion method in Tehran was 26, 23, 22.5, and 16%, respectively. In general, resistance to INH was more common than RIF, SM, and EMB in TehranConclusions: In conclusion, this systematic review summarized the prevalence and distribution of first-line anti-tubercular drug resistance of M. tuberculosis in Iran. Our results suggested that effective strategies to minimize the acquired drug resistance, to control the transmission of resistance and improve the diagnosis measures for TB control in Iran. Mycobacterium tuberculosis in Iran, one of the eastern Mediterranean countries locating between Azerbaijan and Armenia and high-TB burden countries (such as Afghanistan and Pakistan). Since 1996, when the national TB control programs established in Iran, TB incidence has been declining from 34 per 100,000 to 21 per 100,000 cases in 2011 remains as one of the most common infectious disease in developing countries Full text was available. (2) An original article was performed. (3) Susceptibility data for at least one anti- tubercular drug was available. (4) The laboratory method was used.We sought any articles of antimicrobial susceptibility testing of N < 10).Studies with at least one of the following aspects were excluded: (1) Studies that were not relevant. (2) Articles with only available abstracts (without full text). (3) Studies that did not use laboratory methods (using patients records). (4) Articles that use of second line of antimicrobial drug resistance. (5) Articles that were review. (6) Articles which contain no eligible data. (7) Case series reports. (8) Articles that sample size is too small (http://www.equator-network.org).At this stage, articles with the following features were excluded as well: (1) Any articles were published both in English and Persian. . (2) Duplicate publications. For all studies, we extracted the following data from the original publications. Literature identification and data extraction was performed by two researchers independently. Quality assessment of methodological sections and results of included articles was performed by use of STROBE checklist , different types of PCR , MGMT, and MGIT (direct and indirect). But most of them that used were agar proportion or PCR. In all the studies that use both of them, the results of reference method (agar proportion) had the highest of sensitivity and specificity . In Masjedi et al. (This review addressed the prevalence of first-line anti-tubercular drug resistance of i et al. , Bahrmani et al. reportedi et al. . In Mohai et al. reports,i et al. this levi et al. and Merzi et al. study, aM. tuberculosis resistant at least to INH and RIF, was 3%. In Ayaz et al. (In Al-Akhali et al. study thz et al. study thSome limitations of this systematic review should be considered for results interpretation. First, few studies have been conducted in our country about resistance of TB to first and second line-drugs. Second, the probable influence of age, sex, ethnicity, economic level, and life styles could not be analyzed due to the limited information obtained from the original articles. Third, most included studies were hospital-based rather than population based which makes the results more prone to potential selection bias. Because of the small number of studies particularly in other cities except Tehran, we cannot judge about the prevalence of resistance against first-line anti-tuberculosis drugs properly. However, in recent years, emergence and spread of MDR-TB threaten the TB control strategy. In many law-and middle-income countries, due to inadequate laboratory capacity, most of the patients with MDR-TB are not diagnosed. Treatment of these cases mostly failed and significant expenditure of health care resources is needed.M. tuberculosis in Iran. Our results suggest effective strategies to minimize the acquired drug resistance, to control the transmission of resistance and improve the diagnosis measures for TB control in our country.In conclusion, this systematic review summarized the prevalence and distribution of first-line anti-tubercular drug resistance of An important element in gaining control of this epidemic is developing an understanding of the molecular basis of resistance to the most important anti-tuberculosis drugs. Since the mechanism of action of rifampin is to inhibit mycobacterial transcription by targeting DNA-dependent RNA polymerase (Somoskovi et al., On the other hand, INH is activated by the mycobacterial enzyme KatG, a multifunctional catalase-peroxidase that has other activities including peroxynitritase and NADH oxidase. Therefore, inhibition of both cell wall lipid, and nucleic acid synthesis by INH-NAD and INH-NAPD adducts together with respiratory inhibition by INH-derived NO can provides a potent antituberculosis cocktail. Some strategies such as developing agents that produce the isonicotinoyl radical, screening for molecules which increase mycobacterial levels of NAD+ or NADP+ for in co-administration use with INH, to designing of more drug-like molecules using the structure of INH-NAD adducts to inhibit specifically mycobacterial enzymes; and developing of mycobacterial enzyme inhibitors which can inactivate INH might be useful to control INH-TB resistance propagation (Timmins and Deretic, All authors listed, have made substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "In a unique collaboration between The Seventh Hospital of Hangzhou, the International Society for Traumatic Stress Studies (ISTSS), and the Zhejiang Behavior Medicine Association, the international conference \u201cPosttraumatic stress: state-of-the-art research and clinical implications for China\u201d was organized in Hangzhou, China, on 17\u201319 October 2014.The organizational team was led by Dr Zhonglin Tan, a psychiatrist from The Seventh Hospital of Hangzhou, who has spent a year with Professor Olff's research team in Amsterdam, The Netherlands. With the support of Dr Zhang, Director of The Seventh Hospital of Hangzhou, and with guidance from Professor Olff, Dr Zhonglin Tan successfully created a full program. Over 300 participants, the majority of them from China, enjoyed one full day in both English and Chinese (with simultaneous translation) and the rest of the meeting in Chinese only.global meetings program (see www.istss.org). Recognizing that trauma is a global issue (Schnyder, The renowned board members of ISTSS (Kaysen, Stappenbeck, Rhew, & Simpson, Next to the international speakers, a strong representation of Chinese PTSD experts was seen who gave keynotes on topics ranging from biological stress systems (Bao & Swaab, European Journal of Psychotraumatology (Cao, Wang, Wang, Qing, & Zhang, Apart from keynote lectures, this collection contains the abstracts of a selection of over 100 submissions, the authors of which were awarded for their excellent contribution to the field with a certificate and abstract publication in the Miranda OlffEditor-in-ChiefZhonglin TanHangzhou Mental Health CenterHangzhou Seventh Peoples\u2019 HospitalHangzhou, People's Republic of China"} +{"text": "Task Group (TG) 241 of the American Association of Physicists in Medicine (AAPM), operating under the Work Group for Technology Assessment in Image-Guided Interventions, was formed in September of 2013 with an international membership of experts from MR-guided focused ultrasound (MRgFUS), focused ultrasound and ultrasound physics, radiation oncology, industry and federal agencies.The charge of TG-241 is to report through peer-reviewed publications on the assessment of state-of-the-art clinical MRgFUS technology, including the intrinsic system characteristics, quantitative metrics, sources of uncertainty, quality assurance measures, data types and nomenclature as well as training issues for medical physicists.In addition, through related sub-group activities, TG-241 will consider development of an open tool as a public resource for the MRgFUS research community.The goal of this presentation is to introduce TG-241 to the research community and solicit input about its current activities and future directions."} +{"text": "The concept of predictive personalized medicine is becoming one of the long-term priorities for the development of medical science in the structure of global trends. So, the main goal is: Development of educational programs and specialized didactic materials in PPPM for teachers of the universities and departments of post-graduate education, for students and for practical doctors. Program curriculum have to be structured according to the Bologna Process (European Higher Education Area). It is a difficult goal and we have to understand the new philosophy of up-to-date medicine for integration all the modern interdisciplinary innovations and the best researches of traditional eastern schools (Chinese and Ayurvedic). The duration of the Program maybe 2000 - 3000 academic hours , it have to include several modules .Triad of health \u2013 mental, biochemical and structural components of phenotype. The study of health and life-quality biomarkers (\u201ccorridor of self-regulation\u201d), markers of predisposition. Studying the status of healthy persons in order to understand their neuro-autonomic homeostasis, peculiarities of constitution and applied physiology. This module have to include express - diagnosis methods of visualization: optimal body posture, reflected zones of internal organs \u2013 \u201ctissue substrates\u201d in normal and in pathological condition; criteria of optimal cerebral metabolism, lymph circulation etc. New protocols.Stress and General Adaptation Syndrome. Emotional, oxidative, muscle, postural stress - early preclinical diagnosis of stress-induced conditions \u2013 non-specific grain for clinic behavior of multifactorial inflammatory and degenerative diseases.Genetic testing of personal profiles: ONCO-gen, OSTEO-gen, CARDIO-gen, NEURO - gen, WEIGHT-gen etc. and correlation monitoring with clinical status in groups of risk. Learning the provoking role of stress-factors in the process of disease behavior. Epigenetic medical research - genetics of interaction, study of environmental and internal factors, ethics, culture and personal responsibility of an individual for his or her own health.Creation of individual profiles in PPP programs, including conventional methods and medicines. Motivation and recommendations in PPPM for patients. Non-drug therapies for detoxification, correction of posture, cerebral metabolism, personalized exercises, intermitted hypoxia therapy etc.The education program needs to have 4-5 basic modules and every EPMA section may prepare special modules to accumulate the up-to-day knowledge and leading technologies with the purpose of creating interdisciplinary programs for maintaining the spiritual, psycho-emotional and physical health."} +{"text": "There are errors in the funding statement. Please see the correct funding statement below.MCT and FB are supported by the European Union , JCG by a grant of the Ministerio de Econom\u00eda y Competitividad, Instituto de Salud Carlos III, Spain, (PI12/00567), and European Union . JMGA is supported by a fellowship from the Regional Government of Madrid in Spain (PROMT-S2010/BMD2414).The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The complete, correct Funding Statement is as follows: This study was supported by the Czech Ministry of Health, NS10336-3/2009, The publisher apologizes for the errors."} +{"text": "In our recent paper, Somatic Experiencing: Using interoception and proprioception as core elements of trauma therapy SE-based intervention [Trauma First Aide, developed by Miller-Karas and Leitch , and nowSomatic Therapy Treatment Effects with Tsunami Survivors.Parker et al. : SomaticParker presents a similar study of victims of the same tsunami in southern India. A 75-min SE-based intervention was provided to 150 participants with symptoms of trauma. Several outcome measures were taken at immediate post, 4-week and 8-month follow-up, with significant results indicating substantial benefit. At intake, 80% or participants had one or more PTSD symptoms of arousal and intrusion, and 50% had avoidance symptoms; at 8 months follow-up, 90% had significant or complete improvement.Somatic Experiencing treatment with Social Service workers following hurricanes Katrina and Rita.Leitch et al. : SomaticThis paper describes using 1 or 2 sessions of TRM with Social Service workers in the aftermath of hurricanes Katrina and Rita. The treatment group showed significant reduction in PTSD symptoms and increased resilience at 3\u20134 months follow-up.A case for using biologically-based mental health intervention in post-earthquake China: Evaluation of training in the trauma resiliency model.Leitch and Miller-Karas : A case This paper documents the provision of TRM training to 350 disaster responders in Sichuan province, China, after the 2008 earthquake. Ninety seven percent of respondents believed the training would be moderately to very useful in their work.Mindfulness-Based Mind Fitness Training: A Case Study of a High-Stress Predeployment Military Cohort.Stanley et al. : MindfulStanley presents an outcome study of Mindfulness-Based Mind Fitness Training (MMFT), derived from SE, TRM, and Mindfulness, with a group of 34 Marine reservists. Increased mindfulness correlated with time spent practicing and with reduced stress.Stress is Dominant in Patients with Depression and Chronic Low Back Pain.Ellegaard and Pedersen : Stress Ellegard offers a qualitative study, using a phenomenological-hermeneutic approach, of 6 patients with non-specific low back pain receiving Gestalt Therapy and SE. The study does not enable a separation of the effects of Gestalt Therapy from SE.Clinical Implications of Neuroscience Research in PTSD;Van der Kolk : ClinicaEncoding States: A Model for the Origin and Treatment of Complex Psychogenic Pain;Ruden : EncodinWhole brain integration in the clinical application of Somatic Experiencing.Hricko : Whole bThese studies review aspects of neuroscience supportive of the SE approach, and offer conceptual models similar to our own Levine, . AlthougTaken together, these papers offer evidence supporting continued research into SE. The papers on disaster response in particular, although not definitive, are strongly suggestive of the efficacy of SE as an early, low-dose, culturally flexible intervention for victims and providers in the context of natural disasters.Peter Payne is an SE practitioner (SEP) who derives income from his practice. Peter A. Levine declares that teaching, royalties and consulting related to SE are a source of income. Mardi A. Crane-Godreau is an SEP and non-paid member of the Board of Directors of the Somatic Experiencing Trauma Institute\u2122."} +{"text": "The evolution of AIDS cases in Constan\u021ba County during 1985-2003 shows a rapid upward detection curve in children in the first decade, a low rate of screening in adults in the first decade, with absolute predominance of cases in children and progressively slower increase in the number of cases identified in the second decade, on account of the increase in the number of cases in adults. The main objective of this paper is to present the retrospective and prospective HIV-AIDS epidemiological model.Organization and data collection was performed, through analysis in terms of ordering data, correlation, and validation: descriptive analysis of the elements that determine the distribution of cases. We have identified characteristics from: 1. Time - tendency (age pyramid); 2. Place - geographic , geological, industrial , agriculture, climatic, geopolitical conditions, prior to 1989, blood products, pollution status, place of work ; 3. We evaluated the determinants of public health problems - variable 'person' . Aspects of demography - biological epidemiological indicators like gender/sex , age , ethnicity, race, area of origin , village of origin, place of residence, behaviors , profession, occupation, life style, socio-economic level, and other information - migration, tourism) We proposed to observe, to describe cumulative data, the explanation of the phenomenon, to monitor, to control and to formulate the considerations based on epidemiological verified data.In Constan\u021ba, the top of the curve-infection was 1988, possible transfusion . Constan\u021ba - first certified center in Romania, first family foster homes and palliative therapy center, the highest percentage (99%) of testing of pregnant women. Healthcare system in Constan\u021ba always has anticipated the national Healthcare System development . The tracking system through clinical indicators to monitor and evaluate (July 2009). Initially management of a pioneer public-private partner relationship with Baylor Constan\u021ba is methodological center.The role of epidemiologist has been required - the clinician community! The role of the multidisciplinary team is relevant. Establishment of epidemiological valuable databases on the environmental impact of HIV can be a model for monitoring other chronic infections ."} +{"text": "Family therapy and family-based treatment has been commonly applied in children and adolescents in mental health care and has been proven to be effective. There is an increased interest in economic evaluations of these, often expensive, interventions. The aim of this systematic review is to summarize and evaluate the evidence on cost-effectiveness of family/family-based therapy for externalizing disorders, substance use disorders and delinquency.A systematic literature search was performed in PubMed, Education Resource information Centre (ERIC), Psycinfo and Cochrane reviews including studies conducted after 1990 and before the first of August of 2013. Full economic evaluations investigating family/family-based interventions for adolescents between 10 and 20 years treated for substance use disorders, delinquency or externalizing disorders were included.Seven hundred thirty-one articles met the search criteria and 51 studies were initially selected. The final selection resulted in the inclusion of 11 studies. The quality of these studies was assessed. Within the identified studies, there was great variation in the specific type of family/family-based interventions and disorders. According to the outcomes of the checklists, the overall quality of the economic evaluations was low. Results varied by study. Due to the variations in setting, design and outcome it was not feasible to pool results using a meta-analysis.The quality of the identified economic evaluations of family/family-based therapy for treatment of externalizing disorders, adolescent substance use disorders and delinquency was insufficient to determine the cost-effectiveness. Although commonly applied, family/family-based therapy is costly and more research of higher quality is needed. Family therapy and family-based treatment is considered an evidence-based practice treatment for children and adolescents with externalizing disorders, symptoms of delinquency and/or substance use disorder , 2. FamiFor the purpose of the present paper, family therapy and family-based treatment is broadly defined as treatments in which primarily family members and/or members of the families\u2019 wider networks are involved in the treatment process of resolving problems for young people as opposWell-known forms of family/family-based treatments are Multisystemic therapy (MST) , FunctioRecently, Von Sydow et al. systematCurrent health care policy in the Netherlands and elsewhere places emphasis on the provision of effective mental health services in a cost effective way. Family/family-based interventions are intensive as they consist of a relatively high number of sessions per week and subsequently are relatively expensive \u201316. TherHence, this paper presents a systematic review of economic evaluations of systemic interventions in adolescents with externalizing disorders, substance abuse or delinquency. The aim of the present study was to assess the evidence on cost-effectiveness of family/family-based therapy for adolescents with externalizing disorders, substance use disorders or delinquency, and to evaluate the quality of the existing studies, and the generalizability of the study findings.The review was performed according to the Cochrane handbook for systematic reviews of interventions and adopA systematic literature search was performed in Pubmed, ERIC, Psycinfo and Cochrane reviews . These different search engines were used because of their high quality, coverage of large databases and their focus on economic trials.Search terms encompassed the different types of systemic therapy but also more general classifications . These terms were searched for in title and abstract and were then combined with terms referring to economic evaluations searched for in title and abstract or a Medical Subject Headings (MeSH) term and in the title (costs). Costs were searched for only in the title, and not in the abstract, because the latter resulted in many irrelevant studies. This search term was included as we noticed that although in some studies both costs and effects were evaluated, the main focus of these studies was to evaluate the costs and a smaller part was referring to the effects. Consequently, when only terms referring to both the costs and effects were included, these studies would have been missed. The search term \u201cEconomic modeling\u201d was not explicitly incorporated into the search strategy as the modeling should be part of a cost-effectiveness, cost utility, cost benefit or cost analysis (corresponding with our aim).Abbreviations were also included. To improve our search, MeSH terms were used, see In Fig.\u00a0The quality of the studies was assessed with the British Medical Journal Checklist for authors and peer reviewers of economic submissions and the A total of 731 articles met the search criteria. After removal of duplicates and a first selection based on the abstracts, 51 studies matched the inclusion criteria. After assessment for eligibility, 11 studies were selected . More complex models, like multilevel analysis, should be used. In this way covariates can be included, correlation between measurements over time can be addressed, missing data is accounted for and skewness in the costs and effects is considered. Uncertainty around costs should also be presented by using for instance bootstrapped costs/effects confidence intervals and can be visualized in a cost-effectiveness plane. Sensitivity analysis should be applied to variables that are uncertain . A one way sensitivity analysis is not always sufficient and a sensitivity anaysis also taking into account interactions between variables should be considered. A common discount rate should be applied for all costs and effects. Summary measures of the cost-benefit, cost-effectiveness or cost utility should be given. In case of a cost-effectiveness analysis incremental cost-effectiveness ratio (ICERS) should be calculated. For conducting economic evaluations it is advised to consult a health economist.Although family/family-based treatments are widely used and can be considered as effective for the treatment of a wide range of disorders , cost-efACER, average cost-effectiveness ratio; ACRA, adolescent community reinforcement approach; AT, after treatment; C, comparator; CAU, care as usual; CBCL, child behavior checklist; CD, conduct disorder; CHEC, Consensus on Health Economic Criteria; CM, contingency management; DC, drug court; ENG, England; ERIC, Education Resource information Centre; FC, family court with community services; FFT, functional family therapy; FSN, family support network; Group, skill-focused psycho-education group intervention; GSI, global severity index; I, intervention; ICER, incremental cost-effectiveness ratio; IT, individual treatment; Joint, combination of individual and family therapy; MDFT, multidimensional family therapy; MeSH, Medical Subject Headings; MET/CBT12, motivational enhancement treatment/cognitive behavior therapy, 12 sessions; MET/CBT5, motivational enhancement treatment/cognitive behavior therapy, 5 sessions; MEX, Mexico v; MST, multisystemic therapy; MST-PSB, multisystemic therapy for problem sexual behavior; NA, not applicable; NC, not clear; NHS EED, NHS Economic Evaluation Database; NS, not stated; NS1, reference to non-accessible article; PC, psychiatric crisis; PRISMA, Preferred Reporting for Systematic reviews and Meta-Analyses; RCT, randomized controlled trial; SD, standard deviation; SRD, self-report delinquency scale; SUD, substance use disorder; SW, Sweden; TLFB, timeline follow-back form; UK, United Kingdom; USA, United States of America; USD, United States Dollar; YSR, youth self report"} +{"text": "This editorial deals with the state of the Iranian Journal of Basic Medical Sciences (IJBMS) in 2013. In the previous year, we received 613 manuscripts for publication. After peer-review by expert national and international reviewers, 15 % of submitted papers were accepted for publication. As presented in The publication frequency was doubled by switching from bimonthly to monthly, with around 12 papers per issue. Most of the published papers were original articles; review articles were around 5% of all published papers . Contributions of the international authors were increased compared to the previous years . In 2013Crocus sativus). As Saffron is one the most famous plants cultivated in Iran and has various pharmacological effects including anti-asthmatic, anti-carcinogenic, anti-mutagenic, anti-depressant, and immunomodulating and has antioxidant-like properties, it was selected for a special issue. The second issue was conducted with 20 papers on Human T-cell lymphotropic virus-I (HTLV-I), as this virus is endemic in several regions of the Middle East including Iran; various concepts about this virus and related diseases were discussed.Two special issues were published in 2013. The first one focused on Saffron is expected for the current year."} +{"text": "There is an error in the affiliation for the 11th author. The correct affiliation for Giuseppe Mancia is: Istituto Auxologico Italiano and University of Milano-Bicocca, Milan, Italy."} +{"text": "International Journal of Molecular Sciences accepted nominations for Junior Scientists Travel Awards 2016. Over 300 nominations were received and were evaluated by a panel of judges comprised of International Journal of Molecular Sciences editorial board members.With the goal of recognizing outstanding contributions to the field of molecular sciences by early-career investigators, including assistant professors, postdoctoral students and PhD students, and assisting them in attending international conferences in 2016, last year the We are excited to announce the following winners: Dr. Bert De Rybel, Dr. Jean Christopher Chamcheu, Dr. Jean-Marie Swiecicki, Mr. Pedro Rodrigues, and Dr. Renato Polimanti. They will be supported with up to 800 Swiss Francs each towards their travel expenses to attend international conferences in 2016.Dr. Bert De RybelScience, Nature, Nature Chemical Biology and Current Biology. He then moved to the laboratory of Professor Dolf Weijers in Wageningen (the Netherlands), funded by FEBS and Marie Curie post-doctoral fellowships, to work on embryo development and initiated work into vascular development. After five years of post-doctoral research in which he published his work in leading journals including Science and Developmental Cell, Dr. De Rybel received an FWO Odysseus return grant to set up his own line of research at the department of Plant Systems Biology in VIB-Ghent University, Belgium, where he currently works as a project leader on vascular development in plants. At the same time, he remains associated to Wageningen University, funded by an NWO VIDI grant. His current research focuses on understanding how plant vascular cells control the orientation of their cell divisions and how this has developed through evolution. When not in the lab, Dr. De Rybel enjoys Belgian beers, painting and hiking/camping.Dr. Bert De Rybel, Project Leader at VIB-Ghent University, Belgium. Dr. De Rybel completed his Ph.D. degree in the Department of Plant Systems Biology , supervised by Professor Tom Beeckman, focusing on early lateral root development. This resulted in publications in several leading journals such as Dr. Jean Christopher ChamcheuDr. Jean Christopher Chamcheu, Assistant Scientist at University of Wisconsin (UW)-Madison, Madison, WI, USA. Dr. Chamcheu earned his BS degree in Biochemistry with a minor in Biomedical Analyses and Pharmacology at the University of Dschang, Cameroon, and his MS degree in Biomedicine (2004), at Linkoping University in Sweden. He completed his Ph.D. degree in Dermatology and Venereology (2010) at Uppsala University in Sweden, working with Professors Hans Torma and Anders Vahlquist, to identify the underlying molecular genetic basis and develop new pharmacologic strategies for treating epidermolytic keratinopathic genodermatoses. On completion of his Ph.D., Dr. Chamcheu joined the Department of Dermatology at the UW-Madison as a Postdoctoral Research Associate, working with Professor Hasan Mukhtar for over 3.5 years. His initial research focused on defining the role of Caspase-14 in the pathogenesis of psoriasis, a chronic immune-mediated inflammatory skin disease, and in establishing the utility of delphinidin, a novel small molecule for treating psoriasis. In 2014, he was promoted to the scientist track, where he currently works as an Assistant Scientist, a position at UW-Madison . Dr. Chamcheu is now developing his research program in psoriasis with studies aimed at uncovering the underlying and regulatory mechanisms and contribution of PI3K/Akt/mTOR signaling to psoriasis pathogenesis, and its targeting, using natural small molecule inhibitors with the hope to identify novel prognostic markers and therapeutic targets for treating psoriasis and possibly other hyperproliferative and inflammatory skin disorders. Dr. Chamcheu has recently received a number of grants and awards as co-investigator or as principal investigator, including an American Skin Association Carson Research Scholar Award in Psoriasis. When Dr. Chamcheu is not in the lab, he enjoys outdoor activities, such as playing football soccer, and participates in community outreach organizations, such as the Powerman HOPE Foundation International.Dr. Jean-Marie SwiecickiN-linked protein glycosylation pathway of the human pathogen bacterium, Campylobacter jejuni. When not in the lab, Dr. Swiecicki enjoys baking at home, sailing on the Charles River and discussing politics.Dr. Jean-Marie Swiecicki, Post-Doctoral Research Associate at Massachusetts Institute of Technology, USA. Dr. Swiecicki completed his undergraduate studies in 2011 at the \u00c9cole Normale Sup\u00e9rieure in Paris with a major in Chemistry and a minor in Biology. He then obtained his Ph.D. degree from the University Pierre and Marie Curie in 2014 working under the guidance of Professor Solange Lavielle. His graduate research focused on the development of original strategies to interrogate cellular internalization mechanisms and the subsequent intracellular distribution of cell-penetrating peptides. His studies provided a comprehensive dynamic picture of the internalization processes, which he then used to rationally design a new delivery vector. Continuing on his academic path to decipher the elementary principles of membrane-associated processes, he is now working in Prof. Barbara Imperiali\u2019s laboratory at MIT. As a post-doctoral associate, he is developing an integrated platform for characterizing molecular interactions among associated peripheral and integral membrane proteins, as well as the membrane lipids environment. He is currently applying this platform to the membrane-bound enzymes of the Mr. Pedro RodriguesMr. Pedro Rodrigues, Ph.D. Student at Research Institute for Medicines (iMed. ULisboa), Portugal. Pedro received his BS degree in Molecular and Cellular Biology from the New University of Lisbon in 2011 and a post-graduation in Biopharmaceutical Sciences in 2012. Since beginning his Ph.D. studies in 2013, Pedro has investigated the role of microRNAs and nuclear receptors in non-alcoholic fatty liver disease (NAFLD) pathogenesis, under the supervision of Dr. Rui Castro and Prof. Cec\u00edlia Rodrigues. NAFLD can lead to cirrhosis and hepatocellular carcinoma which lack pharmacological therapeutic options. Studies performed by Pedro have already highlighted novel molecular therapeutic targets that may be useful in developing new drugs to treat NAFLD. He has presented his work in several national and international conferences and is the author of several scientific papers. Pedro has also received several grants and honors, including a National Scholar Award at UEG Week 2014, and his work has been distinguished at several conferences. Outside the lab, Pedro likes to play the clarinet and is a member of several bands and orchestras in Lisbon and in his hometown, Tomar.Dr. Renato Polimantiet al., 2015, Pharmacogenomics) and the molecular mechanisms by which alcohol dependence affects body mass regulation . He is an author of over 50 scientific articles published in international peer-reviewed journals. Dr. Polimanti has received several honors and awards including the NARSAD Young Investigator Award from Brain and Behavior Research Foundation and the Early-Career Investigator Travel Award from the Society of Biological Psychiatry.Dr. Renato Polimanti, Post-Doctoral Research Associate at Yale University School of Medicine and VA CT Healthcare Center, USA. Dr. Polimanti received his Ph.D. degree from the University of Rome \u201cTor Vergata\u201d, Italy, where he studied the evolution of pharmacogenetic systems among human populations. Since 2013, he has been a postdoctoral associate in the group of Dr. Joel Gelernter in the Division of Human Genetics of the Department of Psychiatry at Yale University School of Medicine. Dr. Polimanti\u2019s research is currently focused on the understanding of the pathogenic mechanisms of traits related to alcohol and drug use disorders using various types of genomic data. His recent studies provided novel findings about the role of ancestry genomic background in genome-wide association studies of substance dependencies (Polimanti International Journal of Molecular Sciences\u2019 editorial board members and editorial staffs, we wish to congratulate these five outstanding junior scientists for their accomplishments.On behalf of the"} +{"text": "Human Diseases from Wildlife. Michael R. Conover, and Rosanna M. Vail. ISBN: 978-1-4665-6214-1Rickettsia, viruses, fungi, prions, and parasites). There are then multiple chapters in each section to address specific diseases, 30 in total. Each chapter follows a set pattern of topics . This format provides a simple and easy to follow reference for each topic. The appendices address medical definitions, scientific names of animal species, photos of North American vectors and disease they transmit, and a list of zoonotic diseases covered in this book.Michael Conover and Rosanna Vail are to be commended for compiling the information in Human Diseases from Wildlife, CRC Press (ISBN: 978-1-4665-6214-1). In the true spirit of One Health, this book will prove to be an asset in many collections. The book is 527 pages, including four appendices and a lengthy topical index. Following an introductory chapter, the text is divided into seven sections based on disease-causing agents is limited. For parasites, only baylisascariasis, trichinellosis, and swimmers' itch/giardiasis are addressed.As noted, this book provides a quick reference to field biologists, students, and health care providers to the basics of each of the extensive list of diseases. This book is largely focused, although not exclusively, upon North American wildlife. It is not extremely detailed, but provides basic information, and will be an ideal \u201cfirst stop\u201d reference. Since the book is organized by agents and not syndromes or manifestations, readers will need to know what disease they wish to review. One of the highlights of this text are the multiple \u201csidebars\u201d found within each chapter. These provide anecdotal stories about the various diseases and their recent presentations and relevance/impact upon people and animals. Also, each chapter begins with one or several interesting quotes concerning the diseases. A disappointment in the book is the images. All are black and white and not of the highest quality. Most figures and tables significantly add to the textual presentations. Although the bacterial and viral disease chapters are quite extensive, the coverage of fungal diseases (only All-in-all, I am pleased to have this book in my collection. It will be an asset for One health-related courses, and serve as a ready reference for students wishing to know more about zoonotic diseases associated with wildlife."} +{"text": "Dr. Zhidong Ye is not included in the author byline. He should be listed as the seventh author and affiliated with the Department of Cardiovascular Surgery, China-Japan Friendship Hospital, Beijing, China. The contributions of this author are as follows: Performed the experiments, analyzed the data, and contributed reagents/materials/analysis tools.The incorrect version of"} +{"text": "An affiliation for the last author is not indicated. Vuyiseka Dubula is also affiliated with: University of KwaZulu-Natal, Durban, South Africa."} +{"text": "This work was also supported by the Vienna Science and Technology Fund (WWTF), Proj.Nr. LS11-057 (www.wwtf.at), and the Wings for Life Spinal Cord Research Foundation (WfL), Proj.Nr. WFL-AT-007/11 (http://www.wingsforlife.com). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.A funder is missing from the manuscript\u2019s Funding section. The correct funding statement is as follows: JLVL was supported by the Mexican Council of Research and Technology (CONACYT) grant: 236850 ("} +{"text": "The contents of the DDBJ databases are shared with the US National Center for Biotechnology Information (NCBI) and the European Bioinformatics Institute (EBI) within the framework of the International Nucleotide Sequence Database Collaboration (INSDC). Since 2013, the DDBJ Center has been operating the Japanese Genotype-phenotype Archive (JGA) in collaboration with the National Bioscience Database Center (NBDC) in Japan. In addition, the DDBJ Center develops semantic web technologies for data integration and sharing in collaboration with the Database Center for Life Science (DBCLS) in Japan. This paper briefly reports on the activities of the DDBJ Center over the past year including submissions to databases and improvements in our services for data retrieval, analysis, and integration.The DNA Data Bank of Japan Center (DDBJ Center; To acceg.ac.jp) \u20137. This g.ac.jp) .http://trace.ddbj.nig.ac.jp/jga) has been launched in collaboration with our partner institute, the National Bioscience Database Center of the Japan Science and Technology Agency (http://humandbs.biosciencedbc.jp/en/guidelines) and reviews data submission and usage requests from researchers.Since 2013, the Japanese Genotype\u2013phenotype Archive to develop semantic web technologies for data integration and sharing. We list these achievements independently in the following sections. All resources described here are available from http://www.ddbj.nig.ac.jp and most of the archive data can be downloaded at ftp://ftp.ddbj.nig.ac.jp/.In this article, we report on submissions and updates to the DDBJ databases during the past year, and introduce our services briefly. In addition, this paper also introduces the active collaboration with the Database Center for Life Science and the Korean Intellectual Property Office . The JPO transferred its data to the DDBJ directly, whereas the KIPO transferred its data via an arrangement with the Korean Bioinformation Center (KOBIC). The DDBJ contributed 18.39% of the entries and 11.80% of the total base pairs added to the core nucleotide data of INSD. A detailed statistical breakdown of the number of records is shown on the DDBJ homepage (http://www.ddbj.nig.ac.jp/breakdown_stats/prop_ent-e.html). In addition to the above data, the DDBJ has released a total of 10 765 218 WGS entries (769 genomes), 1 182 612 contig/constructed (CON) entries, 773 TPA entries, 6374 TPA-WGS entries, and 1272 TPA-CON entries as of May 29, 2015. In 2014, most nucleotide data submissions to the DDBJ were made by Japanese research groups and the rest came from India , China , Thailand , Iran , and other countries and regions .Between June 2014 and May 2015, the DDBJ periodical release increased by 11 879 389 entries and 31 427 753 923 base pairs. The periodical release does not include whole-genome shotgun (WGS) and third party data (TPA) files . The DDBRaphanus sativus) genomes submitted by the Kazusa DNA Research Institute; transcriptome shotgun assemblies (TSA) of intestinal metagenomes of grass carp (Ctenopharyngodon idella) submitted by the Institute of Hydrobiology, Chinese Academy of Sciences; expressed sequence tags (EST) of a slime mold (Acytostelium subglobosum) submitted by the University of Tsukuba; eggplant (Solanum melongena) genome submitted by the Kazusa DNA Research Institute; TSA of Raphanus sativus var. sativus, Brassica rapa subsp. pekinensis and their hybrid submitted by the Seoul National University; genome and TSA of two varieties of hops (Humulus lupulus) submitted by the Suntory Global Innovation Center Limited; two swallowtail butterflies, common Mormon (Papilio polytes) and Asian swallowtail (Papilio xuthus), genomes submitted by the University of Tokyo; genomes and transcriptomes of three ants, Wasmannia auropunctata, Monomorium pharaonis, and Vollenhovia emeryi submitted by the Okinawa Institute of Science and Technology; TSA of Welsh onion (Allium fistulosum) submitted by the Institute of Vegetables and Tea Science; Antarctic minke whale genome submitted by the Kyoto University; genomes of two cultivars of a species closely related to sweet potato (Ipomoea trifida) submitted by the Kazusa DNA Research Institute; tadpole shrimp (Triops cancriformis) genome submitted by the Keio University; TSA of common iceplant submitted by the Nagoya University; TSA of the Hokkaido salamander (Hynobius retardatus), which had been submitted by Hokkaido University; and genomes of STAP-related cell lines submitted by RIKEN.Notable data sets released from the DDBJ sequence databases are listed in Table https://ddbj.nig.ac.jp/jga/viewer/view/studies) and NBDC websites. To access individual-level data of these public studies, users need to apply data access requests to the NBDC (http://humandbs.biosciencedbc.jp/en/data-use).As of 1 September 2015, JGA has archived 33 studies (8.1 TB) of individual-level human data submitted by Japanese researchers. Archived studies include exome sequence analysis of cancer and other diseases, epigenetic analysis of Hepatitis B virus integration site, and magnetic resonance imaging of brain from bipolar disorder individuals. Submission of these studies has been reviewed and approved by the Data Access Committee (DAC) at NBDC. The summaries of 19 studies are public both on the JGA . Previously, validation tools for MSS were built with Java 7. In February 2015, the tools were re-built with the new version, Java 8, because of the end of public updates for Java 7 after April 2015.For data submission to the traditional DDBJ database, we provide two systems: the Nucleotide Sequence Submission System NSSS; and the In April 2015, we released the enhanced BioProject/BioSample/DRA submission system. This system enables the users to submit a DRA submission referencing submitted but yet un-accessioned BioProjects and BioSample objects; thus, they need not wait for BioProject and BioSample accession numbers before submitting sequencing data to DRA . These nodes are interconnected with InfiniBand QDR/FDR by a complete bisection fat-tree topology. For the massive data analysis, the NIG supercomputer is equipped with 7 PB of the Lustre parallel distributed file system (http://lustre.org), and for archiving of the Sequence Read Archive data, the 5.5 PB MAID system (http://sc.ddbj.nig.ac.jp/index.php/en/en-sysconfig2/en-hardconfig) and memory-intensive tasks including dconfig) . The numhttp://sc.ddbj.nig.ac.jp/index.php/ja-avail-oss).NIG operates the supercomputer facilities for the purpose of (i) construction and archiving the DDBJ databases, and providing analysis services on them (ii) making research and educational resources available to life science researchers in Japan. For the convenience of the login users, many popular tools and libraries in the bioinformatics domain were installed in the system, as shown on the home page (http://sc.ddbj.nig.ac.jp/index.php/ja-availavle-dbs).In order to help reproduce previously executed analysis flow, different versions of the analytical tools are installed in different search paths. Pre-installed datasets in the NIG supercomputer for those analytical tools are listed on the webpage (http://www.ncbi.nlm.nih.gov/tools/vecscreen/univec) services, which receive requests from web interfaces. The DDBJ Center also provides the new version of Web API for Bioinformatics (WABI) \u201319, the s (WABI) ,21, getes (WABI) .http://ddbj.nig.ac.jp/tx_search/) is an NCBI Taxonomy browsing system in the DDBJ. This browsing system allows data submitters to find authentic scientific names used in the INSDC for the purpose of vocabulary control. Due to the replacement of the NIG supercomputer in 2012, we re-implemented most of our services on open source middleware to become accommodated to the new system. The TXSearch system was built on the Apache Solr full text search system and MySQL. The RESTful Web API service is also provided as shown in Figure ftp://ftp.ncbi.nih.gov/pub/taxonomy).TXSearch (http://p.ddbj.nig.ac.jp) is a high-throughput web annotation system of next-generation sequencing reads running on the NIG supercomputer .To improve the reusability of the sequence annotation data, we have developed a system to make the DDBJ records into the Resource Description Framework (RDF) version in collaboration with DBCLS ,26. We aIn this report, we introduced updates of the DDBJ data sets, data submissions, and analytical systems during the past year. We plan to develop a unified submission portal website for all database systems, in concert with the replacement of our supercomputing system in every 5 years (the next replacement year is 2017). Especially, the JGA system needs update to efficiently archive and distribute ever-growing volume of human genome sequencing data. In terms of RDF, application software is under development as the Microbial BioSample OWL. The current foci on future enhancements of the computer infrastructure in DDBJ are (i) refinement of management process and security infrastructure for JGA; (ii) provision of a computing infrastructure suitable for developers and data analysts on HPC environment; and (iii) performance enhancement of data processing for INSDC database construction and usability. For HPC developers, we are constructing an experimental system for OpenStack private cloud environment on the NIG supercomputer, in addition to the extension of Docker systems for DDBJ analytical services."} +{"text": "Summary: The launch of a new brand and website for DMM. This is part of the gradual implementation of a new Company brand and migration to a new, better, web platform. This culminated in October with the launch of a new website for The Company of Biologists . These small workshops are carefully organised to provide leading experts and early career scientists from a diverse range of scientific backgrounds with an inspiring environment for the cross-fertilisation of interdisciplinary ideas. DMM Editor-in-Chief Ross Cagan is hosting a workshop on \u2018Rethinking Cancer\u2019 in November 2016. The aim of the workshop is to bring cancer experts together with experts in engineering, mathematics and computational biology, entrepreneurs and smart thinkers from the arts to help rethink a challenging and important health problem. We plan to present a series of commentaries and Editorials from the workshop in future issues of DMM. If you would love to host such a meeting, but don't have time to organise it or raise the funding, please contact us (http://www.biologists.com/workshops/propose-new-workshop/). The feedback we receive from attendees and organisers is overwhelmingly positive; you won't regret it!In 2010, the first in an ongoing series of Company-funded workshops took place .Together with Dr Klaus Wagner and Dr Nils Gassen, Natalie was able to complete a co-immunoprecipitation (Co-IP) of the mGluR5/Homer1 linkages in brain tissue from mice subjected to the chronic social defeat stress paradigm. Furthermore, with analysis of the literature and exploratory experimentation, she discovered a new signalling partner with which mGluR5 interacts.in situ hybridisation. Also, she was able to increase her understanding of animal handling and behavioural experimentation, and to join guest lectures to broaden her knowledge.The experience helped Natalie to learn many different experimental techniques, both within and beyond her immediate laboratory group. This included Co-IP, membrane fractionating, western blotting, immunohistochemistry and This opportunity exposed Natalie to new ideas and built the foundation of future collaboration with the Schmidt group at the Max Planck Institute of Psychiatry. It also helped Natalie to secure a post-doctoral position in Munich and the publication of her research.DMM offers Travelling Fellowships to postgraduate students and postdoctoral fellows wishing to make collaborative visits to other research laboratories. Each application is reviewed by the Editor-in-Chief. We receive very positive feedback from recipients, who often find their careers taking exciting new directions following their visit . A zebrafish model of inflammatory lymphangiogenesis. Biology Open4, 1270-1280.Alexandria Voigt, Lida Esfandiary and Cuong Q. Nguyen. (2015). Sexual dimorphism in an animal model of Sj\u00f6gren's syndrome: a potential role for Th17 cells. Biology Open4, 1410-1419.Barbara Dhooghe, Charlotte Bouckaert, Arnaud Capron, Pierre Wallemacq, Teresinha Leal and Sabrina Noel. (2015). Resveratrol increases F508del-CFTR dependent salivary secretion in cystic fibrosis mice. Biology Open4, 929-936.Noriko Wakabayashi-Ito, Rami R. Ajjuri, Benjamin W. Henderson, Olugbenga M. Doherty, Xandra O. Breakefield, Janis M. O'Donnell and Naoto Ito. (2015). Mutant human torsinA, responsible for early-onset dystonia, dominantly suppresses GTPCH expression, dopamine levels and locomotion in Drosophila melanogaster. Biology Open4, 585-595.Colleen Valdez, Reese Scroggs, Rachel Chassen and Lawrence T. Reiter (2015). Variation in Dube3a expression affects neurotransmission at the Drosophila neuromuscular junction. Biology Open4, 776-782.In 2011, discussions among the Directors of The Company of Biologists became focused on the \u2018pain to publish\u2019 experienced by authors \u2013 and whether the Company should do something to help lessen that pain. We conducted a survey of our authors, editors and reviewers; the results sts.org) . BiO is We hope that this Editorial has given you a clearer idea of the ethos and activities of The Company of Biologists. We exist to support biologists and inspire advances in biology. It is this aim that underpins the decisions that the Board of Directors make and the excellence we strive for in our publications. DMM's authors, reviewers and editors are all an essential component of this, and we hope that you will continue to support DMM and The Company of Biologists in the future."} +{"text": "Increased prevalence and widespread use of methamphetamine is the public challenge and worry in the world. It seems that low levels of self-regulation and affective control to carry up probability of psychoactive drugs abuse.The purpose of the present study is the comparison of self-regulation and affective control in methamphetamine and narcotics addicts and non-addicts.In this causative-comparative study, 80 addicts (40 methamphetamine addicts and 40 narcotic addicts) who referred to self-reference quitting addictive centers in Miyaneh, Iran, participated in convenience sampling. Then, they matched up with 40 non-addicts according to age, sex, educational level, and marital status. To collect data, we used self-regulation questionnaire and affective control scale. The data was analyzed by multivariate analysis of variance (MANOVA) and LSD test.Result shows that there is a significant difference between methamphetamine addicts and narcotics addicts and non-addicts in self-regulation and affective control (P = 0.001).This finding indicates that low self-regulation and affective control is a risky factor in psychoactive drugs abuse. The abuse of methamphetamine has increased worldwide in recent years, and methamphetamine use is often associated with psychological disorders . TherefoNowadays, there is an increasing tendency among youth to use methamphetamine, and few research works have been conducted on the importance of the self-regulation and affective control skills of these groups; with emphasis on these findings, educational programs of self-regulation and affective control skills in therapeutic design in therapy centers could be applied. Therefore, the aim of this study is the comparison of self-regulation and affective control in methamphetamine and narcotics addicts and non-addicts.In this causative-comparative study, 80 addicts (40 methamphetamine addicts and 40 narcotic addicts), who referred to self-reference quitting addictive centers in Miyaneh, Iran (2011), participated in the convenience sampling. Then, they were matched up with 40 non-addicts according to age, sex, educational level and marital status. The criteria of selected people included being: male, married, in the age range of 20 to 39 years, and literate. The criteria of exclusion were suffering from: psychotic disorders, bipolar or dissociative disorders; or a severe somatic disease.The questionnaire had 63 items; the subscales to measure the ability to develop, implement, and flexibility to maintain a planned behavior in order to achieve one's goals. Items were answered on a 5-point Likert scale with the following scale points . Scores above 239 indicate high (intact) self-regulation capacity; Scores of 214-238 indicate intermediate (moderate) self-regulation capacity, and scores less than 213 indicate low (impaired) self-regulation capacity. Reliability of the SRQ appears to be excellent. In a community sample of 83 people with varying levels of alcohol problem severity, the SRQ was administered twice, within 48 hours, to test the stability of scores it provides. Test-retest reliability for the total SRQ score was high . The SRQ has also shown strong convergent validity with concomitant measures. In community sample Aubrey et al. (1994), SRQ score was significantly and inversely correlated with volume of alcohol consumption per occasion and with negative consequences of drinking . This means that people with lower scores on the SRQ were more likely to be heavy and problem drinkers .This scale is designed by Williams and Chambless (1992) and it has 42 items and four subscales . To obtain the overall scale score, first convert the responses of reverse worded items, and then compute the average of all 42 responses. Cronbach\u2019s Alpha in the study of Williams and Chambless (1992) was 0.94 in overall scale and varied from 0.72 to 0.91 in the subscales. Test-retest reliability for the total ACS score for two-week test-retest was 0.78 and in the subscales it varied from 0.66 to 0.77. Also, Construct Validity- overall scale discriminant validity with Marlow-Crowne Social Desirability Index obtained -0.17. Emotional Control Questionnaire convergent validity was -0.72 (P = 0.001) . AccordiAfter selecting the research sample, the questionnaires were distributed to the subjects, and they were asked to complete the research instruments. The data was analyzed by multivariate analysis of variance (MANOVA) and LSD test.The results show that 32% of methamphetamine addicts, 21% of narcotic addicts, and 40% of non-addicts covered an age range of 20-29 years old. 68% of methamphetamine addicts, 79% of narcotic addicts, and 60% of non-addicts covered an age rang of 30-39 years old. Also, the results show that 68.6% of methamphetamine addicts, and 55.5% of narcotic addicts had precedent consecutive quitting. According to The aim of the current study was to compare self-regulation and affective control in methamphetamine and narcotic addicts and non-addicts. Results showed that there are significant differences between methamphetamine and narcotic addicts and non-addicts in self-regulation and affective control. In fact, the results showed that methamphetamine addicts in comparison with narcotics addicts and non-addicts; and narcotics addicts in comparison with non-addicts had lower self-regulation and affective control. These results are in line with the outcome of Sussman et al. , Salo et"} +{"text": "There are errors in the paper. Please see below for descriptions of the errors and their corrections.There are errors in the Funding section. The correct funding information is as follows: The research work was supported by Malaysian Palm Oil Board (MPOB) through research grant R000999000-RB01-J. The MPOB Director-General supported publication of the manuscript and did not influence the content.An affiliation for the first author is missing. Pek-Lan Chan is affiliated with #1 Advanced Biotechnology and Breeding Centre, Malaysian Palm Oil Board (MPOB), No. 6, Persiaran Institusi, Bandar Baru Bangi, Kajang, Selangor, Malaysia, and #3 School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, UKM Bangi, Selangor, Malaysia.There are errors in"} +{"text": "Today\u2019s modern research of B and T cell antigen receptors (the immunoglobulins (IG) or antibodies and T cell receptors (TR)) forms the basis for detailed analyses of the human adaptive immune system. For instance, insights in the state of the adaptive immune system provide information that is essentially important in monitoring transplantation processes and the regulation of immune suppressiva. In this context, algorithms and tools are necessary for analyzing the IG and TR diversity on nucleotide as well as on amino acid sequence level, identifying highly proliferated clonotypes, determining the diversity of the cell repertoire found in a sample, comparing different states of the human immune system, and visualizing all relevant information.We here present IMEX, a software framework for the detailed characterization and visualization of the state of human IG and TR repertoires. IMEX offers a broad range of algorithms for statistical analysis of IG and TR data, CDR and V-(D)-J analysis, diversity analysis by calculating the distribution of IG and TR, calculating primer efficiency, and comparing multiple data sets. We use a mathematical model that is able to describe the number of unique clonotypes in a sample taking into account the true number of unique sequences and read errors; we heuristically optimize the parameters of this model. IMEX uses IMGT/HighV-QUEST analysis outputs and includes methods for splitting and merging to enable the submission to this portal and to combine the outputs results, respectively. All calculation results can be visualized and exported.http://bioinformatics.fh-hagenberg.at/immunexplorer/.IMEX is an user-friendly and flexible framework for performing clonality experiments based on CDR and V-(D)-J rearranged regions, diversity analysis, primer efficiency, and various different visualization experiments. Using IMEX, various immunological reactions and alterations can be investigated in detail. IMEX is freely available for Windows and Unix platforms at The online version of this article (doi:10.1186/s12859-015-0687-9) contains supplementary material, which is available to authorized users. IGH and TRB V domains are encoded by three different gene segments: variable (V), diversity (D) and joining (j); IGK, IGL and TRA V domains are encoded by two gene types, V and J ) that can classify immune status of patients with distinct diseases .In addition, we plan to extend our analyses to other IG and TR loci . Medium-term we are aiming to integrate a machine learning approach Project Web-page:http://bioinformatics.fh-hagenberg.at/immunexplorer/Operating System: Windows, Linux and UnixProgramming Language: C#Other requirements: Microsoft.NET framework 4.0License: see License Agreement on IMEX website http://bioinformatics.fh-hagenberg.at/immunexplorer/"} +{"text": "The role of the funder is stated incorrectly in the Financial Disclosure (FD) statement of this article. Please refer to the correct FD statement below:This study was sponsored by sanofi-aventis K.K. Dr. Tetsuya Ohtani is an employee of sanofi-aventis KK and received salary from sanofi-aventis KK. The funder had a role in study design, data collection and analysis, decision to publish, and preparation of the manuscript."} +{"text": "Institut d'Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS), Barcelona, Spainhttp://www.enerca.org) : 1) e-Registry, a Pan European registry of RAs for gathering patient\u2019s data necessary to achieve the required sample size for epidemiological surveillance and clinical research 2) e-Learning , a teaching platform for the dissemination of knowledge, continuous medical education, and best practices awareness and promotion through Internet, and 3) e-Medicine , a platform to provide, at distance, expertise (telexpertise) and diagnostic facilities (telediagnosis), avoiding, when possible, the need of travelling. Finally, e-ENERCA will also promote the recognition of the previously identified Centres of Expertise in RAs (White Book) by the national health authorities, a mandatory condition for ENERCA final recognition as European Reference Network in Rare Anaemias (RA-ERN).Rare Anaemias (RA) are a group of Rare Diseases (RD) with prevalence, in Europe, less than 5 per 10.000 individuals. Major forms of RAs require red blood cell transfusions, iron chelation, splenectomy, and/or in very severe cases, bone marrow transplantation, as main therapeutic options. Beta-thalassaemia major is predominant in Italy and Cyprus, and sickle cell disease (SCD) in African population. During the last 30 years, SCD is increasing in Europe due to African immigration, leading to an important impact on health care burden in several countries. Preventive programs, aiming to epidemiological control, and improvement of diagnosis and clinical management of major RA, are crucial for decreasing the affected birth rate and achieving an efficient balance between morbidity and patient\u2019s life expectancy. Since 2003, the European Network for Rare and Congenital Anaemias (ENERCA) has taken an active role for improving this situation by the following actions: a) the identification of Centres of Expertise on RAs in Europe according to the recommendations of ENERCA White Book b) the promotion of best clinical and laboratory practices by the publication of ENERCA recommendations c) the improving of continuous medical education, by organising topic-specific training courses, workshops and symposia, e) the empowerment of patients, by cooperation with Patient\u2019s Associations, and co-organizing a bi-annual European Symposium on RAs with interactive patients-health professionals sessions. In September 2013, a new phase of the project called e-ENERCA has started with the aim to provide, patients and professionals with e-Health tools for assure the same access to health services in RAs across Europe, independently from the country of practise and origin of the patients. e-Health services will be developed through the set-up of three different e-platforms endorsed by ENERCA website ("} +{"text": "Following the publication of this article, Dr. Marc Lippman from the University of Miami Miller School of Medicine raised concerns that the article was based on work completed in his laboratory and which had been employed by the authors without his knowledge or permission.Dr. Lippman's concerns were evaluated by a review panel at Morehouse School of Medicine which concluded that there was substantial evidence in support of Dr. Lippman's claims and advised that the publication should be retracted. The concerns have also been raised to the attention of the University of Miami which advised proceeding as recommended by Morehouse School of Medicine.In the light of the recommendation of Morehouse School of Medicine, the editors retract this publication.We also wish to make readers aware that references 16 and 21 in the original paper were cited with incorrect author details. The two references relate to the same article and the correct full citation is as below:Correlation of GREB1 mRNA with protein expression in breast cancer: validation of a novel GREB1 monoclonal antibody. Breast Cancer Res Treat. 2010 Jul;122(2):371-80. doi: 10.1007/s10549-009-0584-x. Hnatyszyn HJ, Liu M, Hilger A, Herbert L, Gomez-Fernandez CR, Jorda M, Thomas D, Rae JM, El-Ashry D, Lippman ME."} +{"text": "Rinderpest vaccine strain LA-AKO, which is less virulent especially to highly susceptible Asian cattle breeds, was established from a lapinized vaccine strain by further passages in rabbit and chick embryos. Here, we report the genome sequence of LA-AKO, which currently remains active for the production of an emergent vaccine in Japan. Morbillivirus genus of the Paramyxoviridae family. RPV is highly contagious and causes severe diarrhea with high mortality in cattle, buffaloes, sheep, goats, pigs, and other species. Hence, it has been considered one of the most important pathogens for wild and livestock animals in the world. Due to the implementation of efficient vaccines combined with extensive surveillance led by international initiatives, there has been no reported field case since 2002. In 2011, FAO/OIE finally declared global eradication of rinderpest (Rinderpest virus (RPV) is an enveloped single-stranded negative-sense RNA virus belonging to the The genetic information of RPV will be useful to regenerate viruses in emergent situations in the posteradication era. However, to our knowledge, the present data deposited in public databases lack the information on a currently active vaccine strain, LA-AKO, which was established in Japan kit . The complete genome of LA-AKO was 15,882\u00a0bp in length, which was identical in length to those of previously known RPVs. LA-AKO shows 98% identity with the parental Nakamura III at the nucleotide level and was classified in the same clade of Asian lineage of RPV by phylogenetic analysis (MEGA6) . In the LC057619.The genome sequence of LA-AKO has been submitted to DDBJ/EMBL/GenBank under the accession number"} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This study was supported by a Grant-in-Aid for Scientific Research (B) 20330141, 26285155, Grand-in-Aid for Exploratory Research 26590143 and Grant-in-Aid for Scientific Research on Innovative Areas 26118707 from the Japan Society for the Promotion of Science (JSPS) and JSPS KAKENHI, Grand-in-Aid for JSPS Fellows 242666. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Articles from all past issues are indexed, as will be the current and future issues.The Fall 2015 issue of the International Journal of Telerehabilitation (IJT) presents original articles in the areas of Research; Privacy and Security; and Technology Review. As of May 2015, the International Journal of Telerehabilitation (IJT) is live on PubMed Central: The Fall 2015 issue of the International Journal of Telerehabilitation (IJT) presents the original work of three distinguished and innovative interdisciplinary teams. Interdisciplinarity is also a hallmark of the IJT editorial staff, reviewers, and publishing team.The first article is a product of engineers with expertise in computing science (LoPresti and Simpson) and a medical speech-language pathologist (Jinks) who practices in the area of assistive technology. These authors reported upon the degree to which consumers are satisfied with the provision of telerehabilitation services for augmentative and alternative communication or alternative computer accessibility.A second article, by Proffitt, and Lange, demonstrated the feasibility of employing a 6-week, game-based, in-home telerehabilitation exercise program using the Microsoft Kinect\u00ae for individuals with chronic stroke.Finally, the third article co-authored by Watzlaf , DeAlmeida , Zhou , and Hartman describes a protocol to conduct systematic reviews of research in telerehabilitation, with the aim that IJT readers can ultimately apply this protocol to identify best practices in telerehabilitation.http://www.ncbi.nlm.nih.gov/pmc/journals/2411/. Articles from all past issues are indexed, as will be the current and future issues.The International Journal of Telerehabilitation (IJT) is live on PubMed Central: ecohn@pitt.edu) or Senior Associate Editor Jana Cason if you are interested in contributing to a future issue.The next volume of the International Journal of Telerehabilitation will be published in 2016. We cordially invite your submissions by February 2, 2016. IJT accepts original research, case studies, viewpoints, technology reviews, book reviews, and country reports that detail the current status of telerehabilitation. Our peer reviewers constitute a multi-disciplinary group, and include researchers and clinicians from each of the major rehabilitation disciplines, rehabilitation engineers, health information managers, information technologists, and others. We welcome new peer-reviewers and invite guest editors with ideas for special, thematically focused issues. The IJT publication team is agile and can add additional issues as warranted to ensure currency. Please contact Editor Ellen Cohn, PhD (Respectfully,Ellen R. Cohn, PhD, CCC-SLPIJT EditorJana Cason, DHS, OTR/L, FAOTAIssue Co-Editor and IJT Senior Associate Editor"} +{"text": "Abstracthttp://deepsea.biodiv.tw/)\u201d as part of the \u201cFish Database of Taiwan,\u201d can benefit the study of temporal and spatial changes in distribution and abundance of fish fauna in the context of global deep-sea biodiversity.The study of deep-sea fish fauna is hampered by a lack of data due to the difficulty and high cost incurred in its surveys and collections. Taiwan is situated along the edge of the Eurasia fig, at the junction of three Large Marine Ecosystems or Ecoregions of the East China Sea, South China Sea and the Philippines. As nearly two-thirds of its surrounding marine ecosystems are deep-sea environments, Taiwan is expected to hold a rich diversity of deep-sea fish. However, in the past, no research vessels were employed to collect fish data on site. Only specimens, caught by bottom trawl fishing in the waters hundreds of meters deep and missing precise locality information, were collected from Dasi and Donggang fishing harbors. Began in 2001, with the support of National Science Council, research vessels were made available to take on the task of systematically collecting deep-sea fish specimens and occurrence records in the waters surrounding Taiwan. By the end of 2006, a total of 3,653 specimens, belonging to 26 orders, 88 families, 198 genera and 366 species, were collected in addition to data such as sampling site geographical coordinates and water depth, and fish body length and weight. The information, all accessible from the \u201cDatabase of Taiwan\u2019s Deep-Sea Fauna and Its Distribution ( The top 10 orders are Gadiformes, Myctophiformes, Anguilliformes, Stomiiformes, Ophidiiformes, Pleuronectiformes, Argentiniformes, Perciformes, Beryciformes and Squaliformes , eastern Taiwan, southeastern Taiwan (Western Pacific) and southwestern Taiwan (South China Sea).Sampling description: The research vessels used were \u201cR/V Fishery Researcher I,\u201d \u201cR/V Ocean Researcher I\u201d and \u201cR/V Ocean researcher III.\u201d Constrained by limited cable length, the maximum depth sampled was 4,460 meters. Major equipment used were otter trawl, French type beam trawl of 4 m span, ORE type beam trawl of 3 m span and Isaacs-Kidd midwater trawl (IKMT). Once the nets reached the sea bottom, they were towed for one hour at a ground speed of 1.5\u20132.5 knot for otter trawls and 1.0\u20131.5 knot for others.Quality control description: All the scientific names of fish samples were validated by the updated fish checklist in the \u201cFish Database of Taiwan\u201d or TaiCOL before they were entered into database. Afterward, they were validated again by matching against FishBase and Catalog of Fishes, California Academy of Sciences for further correction. If a specimen was rare or it might belong to an undescribed or new species, it was photographed in fresh and then both the specimen and its tissue sample were catalogued and deposited at the Biodiversity Research Museum of Biodiversity Research Center, Academia Sinica (ASIZP of BRCAS). The latitude and longitude of trawling routes were plotted on Google Maps and outlier detection was conducted.Step1: Sampling locality and water depth were recorded.Step2: Specimens were roughly classified and counted either right on board or when they reached the shore.Step3: Specimens were shipped back to the lab for species identification, body length and weight measurement, and picture taking.Step4: Specimens were fixed in 10% Neutral Buffered Formalin for one month. Next, they were cleaned with water and preserved in 70% alcohol.Project title: Survey of Deep-Sea Fish Diversity by Research Vessels in Taiwan Waters.Personnel: Kwang-Tsao Shao (Project Director), Jack Lin (Software Engineer and Database Manager), Hsin-Ming Yeh, Mao-Yin Lee, Hsuan-Ching Ho, Yun-Chih Liao, Hen-Wei Lin .Funding: Ministry of Science and Technology , Executive Yuan, R.O.C. (Taiwan).PageBreakStudy area descriptions/descriptor: Taiwan is located on the eastern edge of Asian continental shelf. To the west of Taiwan is the shallow Taiwan Strait, to the northeast is the Okinawa Trough , to the east is the complex and diverse Philippine Sea (with deep oceanic trenches), and to the south is the South China Sea . These deep-water environments were where the surveys were carried out.Design description: This study focused on Taiwan\u2019s deep-sea fish fauna, which so far hasn\u2019t been investigated much, and hoped to learn if the fauna varies depending on the sea area, current and water depth. All the specimens caught went through taxonomic identification and had their collection time, water depth and coordinates recorded. A geographic information system (GIS) on their distributions was established in order to provide references for future academic researches as well as resource development, management and assessment. One or several specimens per fish species were selected to have their photos taken in color. Keeping to the Barcode of Life tissue preservation techniques, a small piece of tissue was excised, preserved in 90\u201395% alcohol and stored at BRCAS in liquid nitrogen canisters. Backup tissue samples were also stored at Livestock Research Institute, COA to facilitate the study of molecular biology and genetics later. The voucher specimens and whole fish specimens were deposited at BRCAS. The specimen information was entered into the Fish Database of Taiwan and is freely accessible to all.Dataset description: The dataset includes station number, locality name, water depth, collection date, latitude, longitude, family name, species name and Chinese common name. Since the number of individuals and the weight and length of each specimen are also included, the data can be used to calculate biodiversity indices, K-dominance (A-B-C) curve and community structure analysis by applying various clustering or ordination methods. They can form a good baseline for the time period of 2001-2006. If more data can be collected in the future, comparisons can be performed and the question of whether deep-sea fish diversity is declining under anthropogenic and climate change impacts can be assessed. The collected specimens (voucher and tissue sample) deposited at BRCAS are open to all users for taxonomic and ecological researches so that studies on new species, phylogeny and zoogeography of deep-sea fishes can be published. Some images and morphological data can also be used by the global databases of FishBase, OBIS, GBIF and EOL. Additionally, detailed analyses on the body size of certain species can generate valuable information on the fish\u2019s early life history and its inshore or offshore migration and recruitment.Object name: Darwin Core Archive A Dataset of Deep-Sea Fishes Surveyed in the Waters around TaiwanCharacter encoding: UTF-8Format name: Darwin Core Archive formatPageBreakFormat version: 1.0Distribution:http://taibif.tw/ipt/archive.do?r=deep-sea-fishesPublication date of data: 2014-08-26Language: EnglishMetadata language: EnglishDate of metadata creation: 2014-08-26Hierarchy level: Dataset"} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This work is funded by the National Plan for Sciences and Technology through project # 11-ENV-1960-02, King Saud University, Riyadh, KSA. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The Dutch National Immunisation Programme includes six tetanus toxoid (TT) vaccinations and reaches a high rate of vaccination coverage. In the Netherlands, several guidelines related to tetanus post-exposure prophylaxis (T-PEP) are in place. In 2003, the Dutch Health Council (HC) reviewed the use of T-PEP. The aim of this study is to evaluate whether the HC recommendations have been implemented.We asked 178 Dutch General Practitioner (GP) offices and 60 Emergency Departments (EDs) to participate in a cross-sectional questionnaire study and requested that participating facilities send in the T-PEP guidelines adopted by their practice. The differences, based on categories mentioned in the HC recommendations, between GPs and EDs and the type of T-PEP guidelines adopted were assessed.The response rates for the GPs and EDs were 38% (n\u2009=\u200967) and 70% (n\u2009=\u200942), respectively. 98% percent (n\u2009=\u2009107) of the participants reported having T-PEP guidelines. Of the guidelines described in the survey responses, 28% were consistent with the HC-recommendations, 36% adhered to the guidelines of the College of GPs (CGP), which restricts the use of T-PEP to tetanus prone wounds but for these wounds is in line with the recommendations of the HC. The remaining 36% had adopted other guidelines, most of which can lead to over-prescription of T-PEP. Information on T-PEP was lacking in patients with higher risk vaccination histories.Almost all participants have adopted T-PEP guidelines. Strict adherence to the HC recommendations is low. More than half of GPs have adopted the more restrictive CGP-guideline, which limits T-PEP to tetanus prone wounds. Clostridium tetani and is not contagious. Spores of the tetanus bacillus, which are present in the soil and in the faeces of domestic animals, can enter the body through a wound. Anaerobic conditions can then lead to production of the neurotoxin tetanospasmin, which causes muscle contractions and spasms .In this study, only presence of T-PEP guidelines was examined. Individual adherence of these guidelines was not examined. Previous research in other countries showed that compliance to T-PEP guidelines was poor ,23,24. OThere are some limitations to this study. Facilities without T-PEP guidelines might have declined to participate, resulting in selection bias. Additionally, this study covers only a small sample of all facilities involved in emergency care in the Netherlands; the sampling procedures were not completely random. Therefore, we cannot be certain that the facilities included in the study were a representative sample. Finally, this study only examined whether health care facilities had adopted guidelines as well as which guidelines had been adopted. Compliance with the guidelines at these facilities was not studied; research in other countries suggests that actual adherence to T-PEP guidelines is poor ,23,24.Almost all participating GPs and EDs had adopted guidelines for T-PEP. Forty-one percent of EDs and 28% of GP offices adopted guidelines fully consistent with the HC recommendations. Tetanus awareness is important for GPs and EDs, especially those treating incompletely vaccinated or unvaccinated patients.CGP: College of General Practitioners; ED: Emergency Departments; GP: General Practitioner; HC: Health Council; NIP: National Immunisation Programme; TIG: Anti-tetanus immunoglobulin; T-PEP: Tetanus post-exposure prophylaxis; TT: Tetanus toxoid; WHO: World Health Organization; WMO: Medical Research Involving Human Subjects Act.The authors declare that they have no competing interests.RD participated in the study design, data collection, data analysis and writing of the manuscript; HdM and NvdM were involved in the study design, data analysis and revision of the manuscript; CS, MtW and SH participated in the study design and revision of the manuscript; and TW contributed to the revision of the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2296/15/112/prepub"} +{"text": "However, global comparative reviews to enhance comprehension of how forest management institutions (FMIs) are conceptualized, and the varying determinants of compliance, are lacking. And so too, is there knowledge fragmentation on the methodological approaches which have and should be prioritized in the We review the regional variations in the conceptualization of FMIs, analyze the determinants of compliance with FMIs, and assess the methodological gaps applied in the study of FMIs.A systematic review of 197 empirically conducted studies (491 cases) on FMIs was performed, including a directed content analysis.First, FMIs literature is growing; multi-case and multi-country studies characterize Europe/North America, Africa and Latin America, over Asia. Second, the structure-process conceptualization of FMIs predominates in Asia and Africa. Third, global south regions report high cases of compliance with informal FMIs, while non-compliance was registered for Europe/North America in the formal domain. Finally, mixed-methods approaches have been least employed in the studies so far; while the use of only qualitative methods increased over time, the adoption of only quantitative approaches witnessed a decrease.Future research should empirically ground informality in the institutional set-up of Australia while also valorizing mixed-methods research globally. Crucially, future research should consider multidisciplinary and transdisciplinary approaches to explore the actor and power dimensions of forest management institutions.The online version contains supplementary material available at 10.1007/s10980-022-01577-8. Globally, forests are at a crossroads\u2014characterized by rapid transformation , intermittent is analogous to medium-term/seasonal streams , and perennial relates to streams that flow all through\u2014analogous to more long-term, enduring institutions How varied are the (non)compliance determinants and outcomes of FMIs? (3) How can we conceptually and methodologically advance research on FMIs? To provide answers to these interrogations, we undertake a review of FMIs. The study is inspired by an earlier review conducted in the context of sub-Saharan Africa . This segment also captured forest management institutional compliance with an emphasis on the variations and determinants. The segment on outcomes explored the ecological, economic, socio-cultural, and political outcomes of FMIs. The framework also has a segment which explores methodological approaches employed in the study of FMIs.The systematic review approach right. In all, while the number of publications increased from 2006, the review shows that it witnessed a decline in 2009 and 2012. The growth in the literature is possibly explained by the interest to uncover institutional \u2018relicts\u2019 (in Africa) and rising environmental challenges in Latin America (bushfires and migration). Formal forest management institutions (structure and process) for forest products have received much attention in the 2000s literature. However, significant growth was observed in the literature of the 2010s, which included both formal and informal institutions and concepts such as ecosystem services, sustainable forest management, farm forestry, biodiversity, REDD\u2009+\u2009, and forest certification. This classification increasingly accommodated the use of endogenous and exogenous institutions, as well as state and community-based institutions. However, both classifications have been (mis)construed to represent formal and informal institutions.On the whole, the literature on institutions has grown over the last 15\u00a0yearsFig.\u00a0. This coFrom a structural dimension, institutions have been most conceptualized as structures in Asia and Africa. These predominate the informal structures where Asia and Africa account for 35% and 28%, respectively, of the review\u2019s literature reporting on informal institutional structures. Latin America closely follows them with 22%. Literature from Asia and Africa further dominates in the classification of formal institutional structures with 28% and 25%, respectively. This is closely followed by Europe/North America (23%). Asia and Africa are \u2018pace-setters\u201d in the implementation of new forest management paradigms such as community-based forest management appear in the literature from Africa and Asia, followed by Latin America. The diversity is rooted in the diverse ethnic arrangements which characterize these regions. Africa, is the most ethnically diverse region in the world Fearon . This dikgotla system of land management practiced by the Batswana people compliance. Additionally, the interaction between formal and informal institutions, including the fallouts of colonial influence, led to the multiplicity of institutions. This possibly explains why compliance predominates the literature in the three regions. In Africa, for instance, pre-colonial types of resource use included the royal hunting preserves of the amaZulu and amaSwati people, and the ple Ghai . Furtherple Ghai and Botsple Ghai , all indple Ghai . Europe/ple Ghai . In the ple Ghai .Fig. 4GlIt is important to note that some of the evoked reasons behind (non)compliance still require further investigation. For example, significant contextual variations in peoples\u2019 attitudes and adherence to forest-sector institutions and governance in Asia, Africa, Latin America, and North America are directly linked to the disparities of key underlying and broader factors such as institutional models and policy frameworks for decentralization . The least employed approach is mixed-methods, as only 8% used this approach Table . Overallpartially enduring and/or take the status of customs and values which transcend several generations , s Ostrom . In bothpros and cons linked to the methods employed. This will further inform the application of methodological approaches and instruments for future empirical studies on forest management institutions. Also, multi-country studies, employing mixed-methods approaches are needed to analyze institutions in forest use and management.From a methodological standpoint, further studies should prioritize methods based reviews further justify the need to pay more attention to the management of forests and forest resources . Future methods-based reviews should consider these.Firstly, while forest management institutions literature has witnessed a growth, this is most significant in Africa and Latin America. Secondly, the structure-process conceptualization of institutions predominates in Asia and Africa. Process-wise, studies from Australia surprisingly did not report on a single process-linked institution. This merits further studies which pays attention to the identification of such institutions. The literature also reports on the drift away from informality to the pursuit of more formal, state-sanctioned institutional arrangements in Australia. Thirdly, global south regions\u2014Africa, Asia, and Latin America\u2014report the highest cases of compliance in the informal institutional set-up, while non-compliance was significantly registered for Europe/North America in the formal domain. Fourthly, politico-economic factors significantly influence institutional compliance in Europe/North America, while economic factors shape compliance in Africa. On the whole, ecological, socio-cultural, and demographic factors were reported to less significantly explain compliance with forest management institutions (FMIs). Fifthly, while forest management institutions in Europe/North America significantly contributed to determining politico-economic outcomes, those in Africa and Latin America contributed to positive ecological and negative economic outcomes. Finally, mixed methods approaches have been least prioritized in the study of forest management institutions; in Africa, the sole application of quantitative methods was prioritized. Future research needs to (1) extend the conceptualization of institutions, (2) increase multi-case and multi-country studies on FMIs especially for Asia and Australia, (3) empirically ground informality in the institutional set up of forest management in Australia, (4) establish in detail, the extent of (non)compliance, their spatio-temporal variations, and determinants, and (5) valorize the application of mixed-methods approaches in the study of FMIs across the globe.To define conceptual and methodological pathways for future studies on forest management institutions (FMIs), this study undertakes a systematic review of the literature on FMIs using 197 papers (491 cases). From the study, the following conclusions are plausible: Supplementary file1 (XLSX 108 kb)Supplementary file1 (DOCX 12 kb)Below is the link to the electronic supplementary material."} +{"text": "Time trends in access to smoking cessation support for people with depression or severe mental illness: a cohort study in English primary care. BMJ Open 2021;11:e048341. doi: 10.1136/bmjopen-2020-048341Falcaro M, Osborn D, Hayes J, The authors want to alert the readers about the below changes:One author was removed from the paper at their request, and the patient and public involvement box and the contributors section have been edited to reflect this. The version displayed will be the latest version."} +{"text": "Development and validation of the protocol for the evaluation of voice in patients with hearing impairment (PEV-SHI)\u201d, published in Braz J Otorhinolaryngol. 2020;86:748-62, with DOI number https://doi.org/10.1016/j.bjorl.2019.05.007, in the title of the article, where it reads:In the article \u201cDevelopment and validation of the protocol for the evaluation of voice in patients with hearing impairment (PEV-SHI)It should read:Development and validation of the protocol for the evaluation of voice in subjects with hearing impairment (PEV-SHI)"} +{"text": "Traditional Chinese medicine splenogastric diseases (TCMSDs) are equivalent to digestive system diseases in modern medicine. The forms of clinical evidence of TCMSDs include clinical trials, such as randomized controlled trials (RCTs) and systematic reviews (SRs). SRs mainly rely on manual operations and have the shortcomings of time consumption and low efficiency; therefore, they cannot meet the needs of rapid clinical decision-making. It is urgent to establish a new and smart form of a database to support the progress of SRs. We searched and screened all TCMSD RCT reports, in both Chinese and English, and extracted them into meta-evidence through predesigned structural Microsoft Excel tables. All meta-evidence was imported into an online clinical meta-evidence collection and management system after data quality checking. The meta-evidence database of traditional Chinese medicine (TCM) splenogastric disease (MED-TCMSD) was then tested as a backend of an automatic meta-analysis system. A total of 405 cases of TCMSD RCTs were processed into meta-evidence. The most common diseases were stomach stuffiness disease, epigastralgia, and chronic atrophic gastritis. Banxiaxiexin decoction and its modifications were the most used interventions. More than half of the cases employed TCM in conjunction with regular therapeutics. The top reported outcomes included clinical effects, adverse events, and TCM syndromes. The MED-TCMSD worked well as a part of the automatic meta-analysis system. We developed and tested a new form of clinical evidence, meta-evidence, for automatic SR and fast evidence-based decision-making. As an example of the MED, the MED-TCMSD can improve the production and updating efficiency of the evidence of TCMSDs. The methods of constructing the MED-TCMSD can be further applied to the development of MEDs of other diseases. H. pylori infection-related diseases. Chen [Traditional Chinese medicine splenogastric diseases (TCMSDs) are a variety of diseases and symptoms that are related to gastrointestinal functions. Their clinical incidence rate is relatively high. For example, the prevalence of functional dyspepsia in China is 7%\u201341%, the prevalence of irritable bowel syndrome in China is 5%\u201325% , Chinesees. Chen construces. Chen establises. Chen developees. Chen , 15. TheEvidence-based medicine (EBM) databases can be classified into four types : clinicaIn this study, we explored a kind of clinical evidence database that is highly standardized and structured to meet the needs of automatic systematic reviewing. We named it the meta-evidence database (MED). It is pre-extracted and stored in a machine-readable data format. We constructed a MED of TCM splenogastric disease (MED-TCMSD) and implemented it in a fast evidence-based decision-making system based on ASRs to test its functions and advantages.The literature databases that we referred to were four Chinese databases and four English databases , Web of Science, and Ebsco Medline). Each database was searched separately for TCM splenogastric disease randomized controlled trials (RCTs) without any time limitations. The search terms were RCT, randomized controlled trial, and a group of specific disease names. We identified 12 TCM diseases corresponding to TCM splenogastric disease: epigastralgia, acid regurgitation, epigastric upset, stomach stuffiness disease, vomiting, hiccup disease, dysphagia disease, regurgitation disease, abdominal pain, diarrhea disease, constipation disease, and dysentery.The inclusion and exclusion criteria followed the patient/population, intervention/exposure, comparison/control, outcome, and study design (PICOS) rule . The incThe screening process was carried out independently by two researchers in three steps: (1) Search results were imported and automatically screened in NoteExpress software to remove the duplicates; they were rechecked manually. (2) The abstracts of the papers were examined carefully according to the selection criteria. Most exclusions were carried out for reasons in this step. (3) The last step was to check all the remaining full texts to make the final inclusion. Two authors completed the screening independently and cross-checked the results for correctness.A standardized extraction table can facilitate the extraction of data. We created a data sheet for extraction based on Microsoft Excel powered with Visual Basic for Applications (VBAs). The extraction tables of the clinical evidence were designed for data collection, which can be imported into the database in the batch mode. The VBA code built in the table can enable basic data inspection and verification of the input data. Each study was assigned a unique number to identify while storing and citing. The full texts (PDF) of the included studies are attached for reference.Based on the characteristics of RCTs, the extraction table contained six sections: general information, evidence sources, clinical data, trial design, grouping and interventions, and outcomes. (1) The general information section has 10 fields, such as evidence ID, evidence status, creation time, evidence name, clinical trial register number, data editor, and reviewer. (2) The evidence sources section has 24 fields, such as source type, article title, journal source, author information, author institution, contact information, grant number, and DOI and full-text path or link. (3) The clinical data section has 23 fields, such as diagnosis (both TCM and Western medicine), sample size, age and gender, course of the disease, inclusion criteria, and exclusion criteria. (4) The trial design section has 19 fields, such as study type, randomization type, blindness type, observation time, and risk of bias factors . (5) The grouping and intervention section has 27 fields, such as group name, sample size of each group, age and gender of each group, observation/control type, name of intervention/drug, dosage form, administration, single dose, frequency of administration, and duration of intervention. (6) The outcome section has 19 fields, such as outcome name, endpoint/change value, direction of effect, result data, and data type (continuous or binary). Clinical data extraction requires clinical knowledge of TCM; hence, we trained data extractors before starting the work. We also employed two experienced reviewers to check all input data to ensure completeness and correctness.th Revision of the International Classification of Diseases (ICD-11) [Regarding standardization of the names of diseases, the names of TCMSDs and those of Western medicine were not unified and did not allow us to form a correspondence map. To create a unified standard, the names of diseases diagnosed by TCMSDs were used to frame the names of diseases diagnosed by Western medicine. With reference to the \u201c11(ICD-11) , Classif(ICD-11) ,\u201d we buiMySQL is an opThe initial search returned 25,831 papers: 8,428 in Chinese and 17,403 in English : 834, Web of Science Core Collection: 7,298, and Ebsco Medline: 8,241). Duplicates were filtered by NoteExpress and manually, resulting in 4,734 papers remaining for the next stage. The inclusion and exclusion criteria were applied to the titles and abstracts of the remaining, and 3,118 papers remained for full-text examination. We downloaded them and read their full text carefully, and finally, 1,600 papers of TCMSDs were included. As the main purpose of this paper was to verify the construction of the meta-evidence database, we chose the most recent five years of RCTs for meta-evidence extraction.A total of 405 cases of TCMSD meta-evidence were collected with Excel tables and imported into the MED.In the 405 cases of TCMSDs, the TCM diagnoses were mainly stomach stuffiness disease (125), epigastralgia (103), diarrhea disease (35), and constipation disease (18). These four TCM diagnoses accounted for 69% of all the included meta-evidence. The top five TCM syndromes were syndrome of Yang deficiency in the spleen and stomach (36), syndrome of cold and heat complex (16), syndrome of dampness and heat in the spleen and stomach (15), syndrome of spleen and stomach deficiency (15), and syndrome of disharmony of the liver and stomach (13). By comparing Western medicine diagnoses to TCM diagnoses, we found that the distribution was relatively uniform, and the top five occurrences were chronic atrophic gastritis of unknown etiology (65), functional dyspepsia (63), chronic superficial gastritis of unknown etiology (53), localized epigastric pain (33), and diarrhea (21) .TCM treatments were mainly herb medicines and Chinese patent drugs, such as banxiaxiexin decoction and its modifications, weifuchun tablets. Weiyan decoction, ziyinyangwei decoction, yiweishengjin decoction, etc., are also occasionally used in clinical use. They are usually combined with Western medicines of gastrointestinal motility drugs and prokinetic agents, such as domperidone, omeprazole, and mosapride citrate tablets .For TCMSDs, most of the included studies employed add-on tests (211 articles) or positive drugs as control (182 articles). Placebo control (3 articles), dose control (3 articles), and blank control (2 articles) were less common. (1) With respect to add-on trials, the patients both in the intervention group and control group were treated with the same baseline therapeutics, and the patients in the intervention group were further treated with TCM interventions. (2) Regarding positive controls, patients in the intervention group were treated with TCM, and those in the control group were treated with an effective routine approach.The outcomes of RCTs of TCMSDs include clinical efficacy measures , safety measures , and prognosis measures . (1) Clinical efficacy measures are quantitative indicators that can objectively describe the clinical treatment effects of interventions. (2) TCM syndrome scores can evaluate the severity and function of the human body. (3) Prognostic measures can reflect the development of disease recovery and outcome after intervention .https://www.pymeta.com/cdss/). The fast EBM decision-making system, namely the TCM clinical evidence auto-analysis and visualization platform, was designed for rapid clinical and health decision-making . This system includes the MED, automatic meta-analysis, machine-based evidence-labeling, and result visualization modules, in which the MED has the key role of auto-SR support. Based on the TCM splenogastric disease meta-evidence, this platform allows an interesting and different process of SR Only five years of TCMSD RCT data were included in this sample database. We plan to process the remaining literature in the future. (2) All the extraction work of the meta-evidence was completed by experienced researchers, which consumed substantial manpower and time resources. We are searching for efficient and sustainable solutions, such as computer-aided technologies and crowdsourcing collaboration to replace the current, inefficient manual methods.Rapid decision-making based on automatic SR techniques is an interesting direction of evidence-based medicine development. ASR techniques need to be supported by more efficient databases than the classical forms of EBM databases . Our new EBM database is highly standardized, structured, and machine-readable, which precisely meets all the requirements of ASRs. A sample of this kind of database, MED-TCMSD, was completed, which has a key role in the ASR and rapid decision-making system. We believe that the meta-evidence approach is a good solution for ASR databases, and the method of constructing MED-TCMSD can be further applied in the development of MEDs for other diseases."} +{"text": "Sacubitril/valsartan and angiotensin-converting enzyme inhibitor (ACEI)/angiotensin-receptor blocker (ARB) therapies were reported to affect glycaemic control and the development of diabetes mellitus (DM), but the findings are inconsistent. We examined the evidence for the effects of sacubitril/valsartan and ACEI/ARB in DM by conducting a meta-analysis.Z test were used to determine the overall results and determine the significance of the RR.The Cochrane Central Register of Controlled Trials (The Cochrane Library), Embase, PubMed, and ClinicalTrials.gov were searched for data from randomised clinical trials (RCTs) that evaluated the efficacy of sacubitril/valsartan and ACEI/ARB in patients, as of May 25, 2022. Patients were grouped by their disease background at baseline. The main outcomes were the number of new-onset DM and hypoglycaemia, elevated glycaemia, inadequate DM control, diabetes treatment, and diabetic complications, from baseline to the end of the trials. The risk of bias was assessed using the revised Cochrane risk-of-bias tool for randomized trials (ROB 2). The quality of the evidence was evaluated according to the Recommendations for Assessment, Development, and Evaluation guidelines. The meta-analysis of the incidence of various outcomes was conducted using fixed or random effects models. The results are expressed as binary risk, 95% confidence interval (CI), and relative risk (RR). The Mantel-Haenszel method and p = 0.02) and HFpEF . Compared with placebo, ACEI/ARB treatment did significantly reduce the risk of new-onset DM among all patients and patients with not all-HF (defined as part of the study population having HF at baseline) and HFpEF , diabetes complications among patients with non-HF , and subsequent diabetes treatment among patients with new-onset DM and significantly increased the risk of hypoglycaemia among patients with not all-DM .This study included 31 RCTs and 86,809 subjects. Compared with placebo, sacubitril/valsartan treatment significantly reduced the risk of new-onset DM among all patients , patients with heart failure (HF) , HF with reduced ejection fraction (HFrEF) , and HF with preserved ejection fraction (HFpEF) . In contrast, sacubitril/valsartan treatment significantly increased the risk of hypoglycaemia among all patients , patients with not all-DM (defined as part of the study population having DM at baseline) , and patients with HFpEF . Compared with ACEI/ARB, sacubitril/valsartan treatment significantly increased the risk of hypoglycaemia among patients with HF (RR 1.85, 95% CI 1.12\u20133.06, The results of our study, especially in reducing glycaemia and new-onset DM, revealed that sacubitril/valsartan had a positive effect on the control of glycaemia and the development of DM. ACEI/ARB also had a beneficial effect but the effect was weaker than that of sacubitril/valsartan. The above effects varied across diseases but the evidence was strongest in patients with HF.CRD42022336311.The online version contains supplementary material available at 10.1186/s12916-022-02682-w. Does sacubitril/valsartan or ACEI/ARB have an effect on glycaemia and the development of diabetes?In this meta-analysis of 31 randomised controlled trials that included 86,809 patients, compared with placebo, sacubitril/valsartan treatment significantly reduced the risk of new-onset diabetes and increased the risk of hypoglycaemia ; ACEI/ARB treatment significantly reduced the risk of new-onset diabetes and diabetes complications among patients with non-HF and diabetes treatment among patients with new-onset diabetes and increased the risk of hypoglycaemia among patients with not all-diabetes .Sacubitril/valsartan and ACEI/ARB had a positive effect on the control of glycaemia and the development of diabetes.Diabetes mellitus (DM) is one of the major public health problems in the world today . The latThe role of angiotensin-converting enzyme inhibitors (ACEI)/angiotensin-receptor blockers (ARB) in glycaemic control and the development of diabetes has long been noted, but the mechanisms for improving glucose tolerance and insulin sensitivity by inhibiting the renin-angiotensin system are complex and unclear. Relevant trials \u20138 showedThe results of the PARADIGM-HF trial led to a landmark drug, sacubitril/valsartan, for the treatment of HF. The post hoc analysis of this trial suggested that sacubitril/valsartan treatment improved glycaemic control and conferred additional metabolic benefit . RelatedEvaluating the role of sacubitril/valsartan and ACEI/ARB in DM, especially among patients with DM combined with HF, is a clinically meaningful issue. The purpose of this meta-analysis was to explore the effect of sacubitril/valsartan on glycaemia and the development of DM by analysing DM-related outcomes in randomised controlled trials (RCTs) and provide an updated analysis of the role of ACEI/ARB in treating diabetes. Furthermore, the results of this meta-analysis will help to provide physicians with information related to glycaemia and diabetes for use when treating patients with sacubitril/valsartan or ACEI/ARB.All included studies were published without moral and informed consent disputes.https://www.crd.york.ac.uk/prospero), register number: CRD42022336311. The review method registered and updated in PROSPERO is described in Additional file All procedures strictly followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). The application of this systematic review protocol for registration has been registered in the PROSPERO database , Embase, PubMed, and ClinicalTrials.gov, the four major medical databases containing the majority of medical research literature, as of May 25, 2022. Two reviewers independently performed literature searches using search strategies designed by author RXW name of the trial, author, and registration number; (2) year of publication; (3) number of participants enrolled; (4) characteristics of the participants at baseline, including DM status, age, and gender; (5) the drug used in the control group; (6) study duration; (7) main outcomes; and (8) the number of participants with new-onset DM, hypoglycaemia, hyperglycaemia, inadequate DM control, diabetes treatment, and diabetes complications from baseline to the end of the study.Two reviewers independently extracted data from the RCTs that met the criteria and the guidelines in Cochrane Handbook [Two researchers separately assessed the risk of bias for each qualified trial using the revised Cochrane risk-of-bias tool for randomized trials (ROB 2) and compiled a risk-of-bias table as described in the Handbook .We used the GRADE principles to assess the quality of the evidence in this meta-analysis. The quality of the evidence was graded as very low, low, moderate, or high by measuring the risk of bias, inconsistency, indirectness, imprecision, and publication bias.A number of adverse reactions related to DM listed in the results of trials include new-onset DM, hypoglycaemia, elevated glycaemia, inadequate control DM, diabetes complications, and diabetes treatment, from baseline to the end of the trials. Among these, the number of new-onset DM cases and the remaining indicators reflected the effect of the drug on the development of diabetes and glycaemic control, respectively.I2 was used to assess heterogeneity. An I2 value of > 50% or a corresponding p-value of < 0.05 was considered to indicate heterogeneity among the studies. In that case, we used a random model and performed meta-regression and subgroup analysis. An I2 of \u2264 50% and a corresponding p-value of \u2265 0.05 were considered to indicate no obvious heterogeneity in the results, and a fixed model was used [Z test were used to determine the overall results and the significance of the RR. A p-value of < 0.05 was considered statistically significant. All results were consistent with the PRISMA at baseline. Sensitivity analysis and publication bias detection were performed using Stata, and was used . Due to Funnel plots and Egger\u2019s test were useInitially, 21,836 possible articles or studies were identified, and 2973 possible articles were left after filtering and removing duplicates. The remaining articles were judged by the three researchers according to the inclusion and exclusion criteria. Finally, 31 RCTs including 86,809 subjects were included in the analysis. The flow chart of study selection is shown in Additional file The detailed characteristics of the 31 trials \u20138, 21\u201345The quality assessment of the included studies is presented in Additional file The meta-analysis results and grades of the quality of the evidence are summarised in Table p = 0.32) and patients with not all-HF (defined as part of the study population having HF at baseline) , HF , HFrEF , and HF with preserved ejection fraction (HFpEF) . There was also no difference in the risk of new-onset DM between patients treated with sacubitril/valsartan and ACEI or ARB and HFpEF , as was the comparison of sacubitril/valsartan and ARB treatment, but the increase in the risk of hypoglycaemia among patients with HFrEF was no significant between-group difference, as was the comparison of sacubitril/valsartan and ACEI treatment and patients with no all-HF , HF , HFrEF , and HFpEF . There was also no difference in the risk of elevated glycaemia between patients treated with sacubitril/valsartan and ACEI or ARB , HFrEF , and HFpEF , as were the comparison of sacubitril/valsartan and ACEI or ARB treatment , HFrEF , and HFpEF , as were the comparison of sacubitril/valsartan and ACEI or ARB treatment and patients with not all-HF and those with HFpEF , as were the comparison of ACEI or ARB and placebo treatment, but the reductions in the risk of new-onset DM among patients with non-HF , HF , and HFrEF were no significant between-group difference , but the increase in the risk of hypoglycaemia among all patients , and patients with non-HF , not all-HF , HFpEF , non-DM , DM , type 1 DM (T1DM) , and type 2 DM (T2DM) were no significant between-group difference, as were the comparison of ACEI or ARB and placebo treatment and patients with non-HF , not all-HF , HFpEF , non-DM , no all-DM , DM , T1DM , and T2DM . There was also no difference in the risk of elevated glycaemia between patients treated with ARB and placebo , non-HF , not all-HF , HFpEF , not all-DM , DM , T1DM , and T2DM , as was the comparison of ARB and placebo treatment , as was the comparison of ACEI and placebo treatment, but there were no significant difference in reduction in the risk of diabetes complications among all patients , patients with not all-HF , HFpEF , non-DM , DM , T1DM , and T2DM , as was the comparison of ARB and placebo treatment , patients with HF , HFrEF , and HFpEF , but among patients with not all-HF , there was no significant between-group difference , patients with not all-DM , and those with HFpEF , patients with not all-DM , not all-HF , and HFpEF , but there was no significant between-group difference and patients with not all-DM and HFpEF , but there was no significant between-group difference and patients with not all-DM and HFpEF , but there was no significant between-group difference , but there was no significant between-group difference .Compared with placebo, sacubitril/valsartan treatment significantly reduced the risks of new-onset DM in patients without DM and patients with HF, HFrEF, and HFpEF by 22%, 76%, 76%, and 46%, respectively, and significantly increased the risks of hypoglycaemia among all patients, patients without DM, and patients with HFpEF by 91%, 471%, and 606%, respectively, but reduced the risks of hyperglycaemia, inadequate DM control, diabetes complications, and diabetes treatment by 39%, 71%, 26%, and 32%, respectively, with no statistical difference. The results were similar in the subgroups.Besides, compared with ACEI/ARB, sacubitril/valsartan treatment increased the risks of hypoglycaemia among patients with not all-DM, HF, HFpEF, and the comparison with ARB treatment by 85%, 85%, and 172%, respectively, with statistically significant differences, but reduced the risks of new-onset DM, elevated glycaemia, DM inadequate control, and diabetes complications by 9%, 19%, 27%, and 20%, respectively, with no significant difference. The results were similar in the subgroups.Compared with placebo, ACEI/ARB treatment did significantly reduce the risks of new-onset DM among patients without DM and patients with not all-HF, HFpEF, and the comparison with ACEI or ARB by 15%, 13%, 40%, 21%, and 11%, respectively, diabetes complications among patients with not all-DM (/non-HF), and the comparison with ACEI by 13% and 15%, respectively, diabetes treatment among patients without DM by 30%, and significantly increased the risk of hypoglycaemia among patients with not all-DM by 106%, but reduced the risks of elevated glycaemia, DM inadequate control, and diabetes complications by 11%, 18%, and 11%, respectively, with no statistical difference.Previous trials showed that the use of renin-angiotensin-aldosterone system (RAAS) inhibitor treatment could induce hypoglycaemia, improve blood glucose levels, and reduce the incidence of DM \u201351. The No direct studies have investigated the mechanism by which sacubitril/valsartan affects glycaemia. As a combined inhibitor of RAAS and neprilysin, the main mechanism of sacubitril/valsartan\u2019s effect on glycaemia may be by inhibiting neprilysin. Neprilysin can decompose a variety of vasoactive peptides, including bradykinin, glucagon, glucagon-like peptide-1, insulin-B chain, vasoactive intestinal peptide, and other substances that play certain roles in glycaemia regulation . SacubitOur findings are similar to the post hoc analysis of the PARADIGM-HF study, which found that sacubitril/valsartan treatment significantly reduced the incidence of new-onset DM and increased hypoglycaemic events in patients with HF, suggesting a role for sacubitril/valsartan in controlling the development of diabetes and a possible role in lowering blood glucose levels. In addition, in the majority of cases, compared with ACEI/ARB or placebo, sacubitril/valsartan treatment reduced the incidence of new-onset DM, hyperglycaemia, inadequate DM control, diabetes complications, and diabetes treatment and increased the incidence of hypoglycaemia. These results also reflect the potential effectiveness of sacubitril/valsartan in treating diabetes in people with different co-morbidities, although statistical significance was not reached. Some additional findings were made in this study. Some differences in the effectiveness of treatment according to the study metrics, especially new-onset DM, hypoglycaemia, and hyperglycaemia, were seen in patients with different types of HF treated with sacubitril/valsartan. The biggest difference was in the risk of new-onset DM , hypoglycaemia (RR 3.59 vs RR 1.18), and hyperglycaemia (RR = 0.70 vs RR = 1.84). Sacubitril/valsartan treatment resulted in a higher proportion of hypoglycaemia in the HFpEF than in the HFrEF group (23/3699 vs 19/5040), and the control group data showed that ACEI/ARB treatment lowered the incidence of hypoglycaemia in the HFpEF group compared with the HFrEF group (6/3686 vs 16/5066), which suggested that the difference in the incidence of hypoglycaemia in different types of HF was not directly caused by the type of HF but by sacubitril/valsartan treatment. Furthermore, the proportion of inadequate DM control (0% vs 0.02%) and diabetes complications (0.02% vs 0.04%) was lower, and hypoglycaemia (0.06% vs 0.04%) was higher in the HFpEF than in HFrEF the group. Overall, sacubitril/valsartan treatment may be more effective in controlling glycaemia in patients with HFpEF than in patients with HFrEF. However, it should be noted that there was a lack of data comparing sacubitril/valsartan and placebo in patients with HFrEF.The effect of ACEI/ARB in preventing DM was similar to the results of previous large clinical studies and meta-analyses , 46\u201348, The differences in the subgroup results may be due to the differential effects of sacubitril/valsartan, ACEI/ARB treatment in patients with various background diseases, such as with or without HF and different types of HF. Currently, no progress has been made in studies of effective ways to treat HF and thus improve DM . TherefoWe conducted a reasonable search of the literature and carefully screened the results using strict standards, which resulted in a large study sample size. This was the first meta-analysis of the effect of sacubitril/valsartan and comprehensive, updated analysis of the role of ACEI/ARB in patients with diabetes, which included only RCTs. Most of the studies in this analysis were large multi-centre clinical trials and most of our analyses were derived from the analysis of moderate to high-quality evidence. Hence, the quality of our meta-analysis was high. Our study confirmed the effect of sacubitril/valsartan and comprehensively analysed the role of ACEI/ARB in DM. Sodium-glucose cotransporter 2 inhibitor has become the only anti-diabetes drug that can reduce HF events, and our study may set the stage for whether sacubitril/valsartan or angiotensin-receptor/enkephalinase inhibitors could be used as anti-HF agents for the treatment of diabetes. However, several possible deficiencies should also be noted. Firstly, the metrics we studied were not the main objective of most of the trials, and the lack of clarity in the definitions and measurement of the metrics in most cases may have resulted in the application of different criteria, as well as bias, in our results. Secondly, no standardised definitions were used for HF, which also may have led to some bias in the subgroup analysis. Thirdly, most trials did not match patients and select dosages based on diabetes status, while studies using sacubitril/valsartan were primarily in people with HF, and studies using placebo were primarily in people with not all-HF, and the observation period of the individual studies was short.The results of our study, especially in reducing glycaemia and new-onset DM, revealed that sacubitril/valsartan treatment had a positive effect on the control of glycaemia and the development of DM, and ACEI/ARB also had a beneficial effect but the effect was weaker than that of sacubitril/valsartan. The above effects varied across disease settings and the evidence may have been the strongest in patients with HF. Hence, sacubitril/valsartan has the potential to become an anti-HF drug for the treatment of diabetes. However, the combined use of sacubitril/valsartan, ACEI, or ARB and conventional doses of diabetes medication may increase the incidence of hypoglycaemia and requires further studies. Dose adjustments of insulin or other antihyperglycaemic agents may be needed, especially in patients with HF. In conclusion, the effect, exact mechanism, and population that may benefit from sacubitril/valsartan treatment in DM need to be clarified by further studies. However, these results will bring more information and inspiration to the prevention and treatment of DM.Additional file 1. The review method registered in PROSPERO.Additional file 2: Table S1-S4. Search strategy in PubMed, Embase, Cochrane Central Register of Controlled Trials, and ClinicalTrials.gov. Table S5. Eligibility criteria of included studies. Figure S1. Flow chart of literature search and study selection. Figure S2-S7. Network charts in network meta-analyses. Figure S8-S9. Funnel charts in direct comparisons. Figure S10-S20. Sensitivity analysis charts in direct comparisons. Figure S21-S25. Funnel charts in indirect comparisons.Additional file 3. PRISMA 2020 for Abstracts Checklist.Additional file 4: Figure S1. Methodological quality graph. Figure S2. Methodological quality summary. Figure S3-S33. Forest maps for all meta-analyses."} +{"text": "To study the clinical values and implications for the prognosis of procalcitonin (PCT) combined with C-reactive protein (hs-CRP) in patients with bacterial bloodstream infection. n = 53) and the gram-positive bacteria bloodstream infection group (n = 31). Depending on the prognostic outcome of the participants after 28 days, they were categorized into survival and fatality cohorts. The PCT and hs-CRP levels were compared to explore diagnostic value implications for the prognosis of the cases with bacterial bloodstream infection. One hundred and twenty patients with infection hospitalized from Mar. 2020 to Jun. 2021 were chosen as subjects. All participants were tested for serum PCT, hs-CRP, and blood culture. According to the types of pathogenic bacteria, they were divided into the gram-negative bacteria bloodstream infection group (P < 0.05). There were 27 gram-positive participants and 9 gram-negative cases in the positive cohort. The serum PCT value of gram-negative bacterial infection was greater than that of gram-positive bacterial infection. The value of serum PCT in the gram-negative bacterial infection group was higher than that in the gram-positive bacterial infection group, and the difference was statistically significant (P < 0.05). The areas under the curve (AUCs) of PCT, combination of hs-CRP and PCT, and hs-CRP were 0.946, 0.783, and 0.991, respectively. The combined examination of PCT and hs-CRP was the largest, PCT was the second, and hs-CRP was the lowest. These results indicated that the accuracy of combined detection of PCT and hs-CRP in the diagnostic bloodstream infection was the highest (0.991), followed by PCT (0.946) and the lowest (0.783). The PCT and hs-CRP levels of the survival cohort were lower than those in the death cohort. AUCs of PCT, hs-CRP and PCT, and hs-CRP were 0.848, 0.826, and 0.934, respectively. The combined examination of PCT and hs-CRP was the largest, followed by PCT and hs-CRP. The accuracy of the combination of PCT and hs-CRP was the highest (0.934), followed by PCT (0.848), and the diagnostic accuracy of hs-CRP was the lowest (0.826). Serum PCT and hs-CRP values in the positive cohort were higher than those in the negative cohort. The levels of serum PCT and hs-CRP in pulmonary infection were higher than those in the group with negative cases, and the difference was statistically significant ( There were significant differences in the levels of PCT and CRP between the gram-positive bacteria group and the gram-positive bacteria group. PCT and CRP have high diagnostic values in predicting the short-term prognosis of patients. PCT and CRP assist clinical diagnosis and guide treatment and play a positive role in early treatment and prognosis evaluation of patients. Bloodstream infection is one of the severe infectious diseases in clinic, which seriously threatens the life and health of patients . BloodstThe incidence of BSI has been increasing year by year. Given the high mortality rate of BSI, the length of stay in hospital is longer and the cost of hospitalization is more expensive, to the extent that patients' lives are seriously threatened. Various invasive diagnostic and testing techniques, like trauma and scald, can disrupt the integrity of the body's barrier function, cause a decrease in immune function, and increase the risk of bloodstream infection. In addition, diverse factors, such as prolonged coma, malnutrition, and advanced age, can also increase the prevalence of bloodstream infections. It therefore becomes important and urgent to identify early, rapidly, and accurately the pathogens causing BSI and then to select the appropriate antibiotic treatment.With the rapid development of biomedical engineering technology, blood culture technology is widely used in clinic and is regarded as the \u201cgold standard\u201d for the diagnosis of BSI. However, this technique still has some limitations, such as the relatively long cycle of detection of pathogenic microorganisms , which often leads to the loss of the best opportunity for diagnosis and treatment of BSI patients. Therefore, it is of great clinical significance to find reliable and sensitive biomarkers for early diagnosis of BSI.At present, the main biomarkers that can be used to diagnose BSI are blood routine test (BRE), procalcitonin (PCT), and C-reactive protein (CRP) , 9. AmonOne hundred and twenty patients with infection hospitalized from March 2020 to June 2021 were selected as the objects of study. All patients were tested for serum PCT and hs-CRP and blood culture. Among the 120 patients with infection, there were 62 males and 58 females with ages from 19 to 85 years (mean 45.83 \u00b1 4.23). Our clinic's Professional Conduct Association gave their approval towards this experiment. Every participant provided written informed consent.(1) There was no limitation to the sex of the patient, and the initial disease was caused by pulmonary infection, urinary tract infection, suppurative meningitis, bacillary dysentery, and other infections. Blood culture samples and serum samples were sent for examination at the same time; (2) without cognitive, language, and intellectual impairment and with basic reading and writing ability; (3) the diagnostic criteria of bloodstream infection met the following requirements: fever, chills, and other symptoms; hematological tests find antigenic substances of pathogens; and/or when pathogenic bacteria or fungi were isolated by blood culture, any of the following requirements can be met: (a) the strains of pathogenic bacteria isolated from many times of blood culture are the same, and the results of the drug sensitivity test are the same; (b) the strains of pathogenic bacteria isolated from other infected sites are the same as those of blood culture, and the results of drug sensitivity test are the same; and (c) it was effective to use sensitive antimicrobial agents for the isolated pathogenic bacteria. (4) The clinical data are complete, (5) there was no history of surgery or trauma within 3 months, and (6) there were no autoimmune diseases and hematological diseases.The exclusion criteria are (1) patients with severe heart, liver, and renal insufficiency and malignant tumors, (2) patients who had been treated with corticosteroids for a long time, (3) patients with hematological diseases, (4) patients who had a history of taking antibiotics and hormones within 3 months before entering the group, and (5) patients with a history of organ transplantation and immunosuppressant use.The levels of serum PCT and hs-CRP were detected in all patients, and blood culture was performed. The sensitivity, specificity, and diagnostic accuracy of three kinds of detection of bloodstream infection were compared to determine their value in early clinical diagnosis and prognosis evaluation of bloodstream infection. The PCT determination kit (electrochemiluminescence) is from Roche Diagnostic products (Shanghai) Co., Ltd., using the Roche automatic electrochemiluminescence immunoassay system. The hs-CRP determination kit (scatter turbidimetry) comes from Shenzhen Pumen Technology Co., Ltd., using the PA-990P specific protein analyzer. The blood culture instrument comes from BacT/ALERT3D automatic Mycobacterium culture and identification system (Merieux). For the identification and drug sensitivity of gram-positive bacteria, VITEKGP was used, with drug sensitivity VITEKAST-GP67. For the identification and drug sensitivity of gram-negative bacteria, VITEKGN was used, with drug sensitivity VITEKAST-GN09.On the day of admission and 28 days after the patient's admission, the cubital venous blood was drawn in the early morning on an empty stomach and centrifuged at 3000\u2009r/min for 15\u2009min. The upper serum was separated for testing. The PCT value was detected by the chemiluminescence method, and the kit was purchased from Zhengzhou Antu Bioengineering Co., Ltd. The hs-CRP level was detected by immunoturbidimetry, and the kit was purchased from Shenzhen Pumen Technology Co., Ltd. Analyzer Roche 411.\u03c72 test) was applied to compare the counting data. The measured data were expressed by x \u00b1 s, the t-test was used, and analysis of variance was used to compare multiple-group data. The difference was statistically significant, and the difference was statistically significant (P < 0.05). The area under the receiver operating characteristic (ROC) curve was used to evaluate the clinical diagnostic value of PCT, hs-CRP, and combined detection in bloodstream infection and prognosis of patients.The SPSS22.0 statistical program was used for data analysis. The chi-squared test (P < 0.05). See The values of serum PCT and hs-CRP in the group with positive blood culture were higher than those in the group with negative blood culture, and the difference was statistically significant (P < 0.05). See In the positive blood culture group, the serum PCT value and hs-CRP value of the patients with pulmonary infection were larger than those in the negative blood culture group, and the difference was statistically significant (P < 0.05). See In the blood culture positive group, there were 27 cases of gram-positive bacterial infection and 9 cases of gram-negative bacterial infection. The value of serum PCT in the gram-negative bacterial infection group was higher than that in the gram-positive bacterial infection group (P < 0.05). There was no significant difference in serum hs-CRP between gram-negative bacteria and gram-positive bacteria (P > 0.05). There was no statistical significance in the comparison of serum PCT and hs-CRP values in patients infected with gram-negative bacteria (P > 0.05). There was no statistical significance in the comparison of serum PCT and hs-CRP values in patients infected with gram-positive bacteria (P > 0.05). See Among the infections of gram-positive bacteria, there were 14 cases of Staphylococcus epidermidis infection, 9 cases of Staphylococcus aureus infection, 2 cases of Streptococcus infection, and 2 cases of Enterococcus infection. Among the patients infected with gram-negative bacteria, there were 4 cases of Klebsiella pneumoniae infection, 3 cases of Acinetobacter baumannii infection, and 2 cases of Escherichia coli infection, and the serum PCT value of gram-negative bacterial infection was greater than that of gram-negative bacterial infection, and the difference was statistically significant (P < 0.05). The area under the joint detection curve of PCT and hs-CRP was the largest, PCT was the second, and hs-CRP was the lowest. The accuracy of combined detection of PCT and hs-CRP in the diagnosis of bloodstream infection was the highest (0.991), followed by PCT (0.946), and hs-CRP was the lowest (0.783). See The AUCs of PCT, hs-CRP and PCT, and hs-CRP were 0.946, 0.783, and 0.991, respectively . See Of the 36 patients with bacterial bloodstream infection, 29 survived and 7 died. The serum PCT and hs-CRP values in the survival group were lower than those in the death group, and the difference was statistically significant (P < 0.05). The AUC of combined detection of PCT and hs-CRP was the largest, followed by PCT, and the lowest was hs-CRP. The accuracy of combined detection of PCT and hs-CRP was the highest (up to 0.934), followed by PCT (0.848), and the diagnostic accuracy of hs-CRP was the lowest (0.826). See ROC curve analysis showed that the AUCs of PCT, hs-CRP and PCT, and hs-CRP were 0.848, 0.826, and 0.934, respectively is a general term for septicemia and bacteremia. Septicemia is when bacteria enter the bloodstream and grow, producing large amounts of toxins and metabolites, causing systemic severe infection syndrome. Bacteremia is toxemia in which bacteria enter the bloodstream temporarily but do not cause obvious symptoms of systemic infection. Under certain conditions, bacteremia and septicemia can be converted into each other . The disRecently, a large number of broad-spectrum antibiotics, hormone drugs, and immunosuppressants have been widely used in clinic with the continuous development of invasive diagnosis and treatment technology. C-reactive protein is an acute phase reactive protein, which is mainly released from the liver under the stimulation of interleukin-6 and other cytokines . When thProcalcitonin, a propeptide of calcitonin, has no hormonal activity and is usually produced by thyroid C cells. In healthy people, almost all calcitonin is converted to calcitonin, so its content in serum is generally lower than that of 0.05\u2009ng/ml. When infection occurs, the content of serum procalcitonin changes at any time . When thIn conclusion, there were significant differences in the levels of PCT and CRP between gram-positive bacteria and gram-negative bacteria. PCT and CRP have high diagnostic value in predicting the short-term prognosis of patients. PCT and CRP assist clinical diagnosis and guide treatment and play a positive role in early treatment and prognosis evaluation of patients."} +{"text": "Hassane Pathology Department, Medical Research Institute, National Research Centre, Dokki, Giza, PO 12622, EgyptIn the original published version of this article, co-author Yasmin Abdel-Latif affiliations are incorrect. The correct affiliations can be found below. The authors apologize for the errors. Both the HTML and PDF versions of the article have been updated to correct the errors.The authors declare no conflict of interest."} +{"text": "Treatment resistant depression is common in older adults and treatment is often complicated by medical comorbidities and polypharmacy. Repetitive transcranial magnetic stimulation (rTMS) is a treatment option for this group due to its favorable profile. However, early influential studies suggested that rTMS is less effective in older adults. This evidence remains controversial.Here, we evaluated the rTMS treatment outcomes in a large international multicenter naturalistic cohort of >500 patients comparing older vs. younger adults.We show that older adults, while having similar antidepressant response to younger adults, respond more slowly, which may help to explain differences from earlier studies when the duration of a treatment course was shorter.Such evidence helps to resolve a long-standing controversy in treating older depressed patients with rTMS. Moreover, these findings provide an important data point in the call to revise policy decisions from major insurance providers that have unfairly excluded older adults. Major Depressive Disorder (MDD) is a leading cause of disability worldwide, and commonly affects older adults , University of Iowa Center for Noninvasive Brain Stimulation , and Champalimaud Foundation Neuropsychiatry Unit , from 2000 to 2021, in compliance with each site's local Internal Review Board (IRB) policies for analysis and publication of clinical data. All adult patients treated with DLPFC rTMS for depression at Boston and Iowa centers were considered for inclusion since informed consent for retrospective clinical data inclusion in the current analysis was exempted by local IRBs. For Lisbon center, only patients who signed informed consent were considered. Depression was considered as any major depressive episode in the context of either MDD or bipolar disorder type II, according to the Diagnostic and Statistical Manual of Mental Disorders of 0.05, according to Benjamini and Hochberg . We did not find a difference between the two age groups regarding the percentage of responders (N = 372 vs. 91), 5 (N = 318 vs. 84), and 6 (N = 303 vs. 80). Regarding trajectory of response to treatment, measured according to % reduction of depression severity in self-report scales, we found that severity decreased across time , with depression severity reducing more rapidly among younger than in older adults. Similar results were obtained when controlling for potential confounding effects of sex, TMS device, stimulation side, stimulation protocol , treatment site, year of treatment, and total number of pulses per treatment , and significant effects of age group and the interaction between time and age . When controlling for the potential confounders mentioned above, as well as baseline BDI, similar effects were obtained for time and interaction, but without a significant effect for age group , but no effect of age group or interaction between both time and age group. Similar effects were observed when controlling for confounders, including baseline PHQ-9 baseline BDI and PHQ9 were 29.3 (\u00b10.6) and 17.8 (\u00b10.4), respectively and/or AP-L (apleone@hsl.harvard.edu).The datasets presented in this article are not readily available because it was generated from different participating centers. Requests to access the datasets should be directed to AJO-M . Champalimaud Foundation patients/participants provided their written informed consent to participate in this study. Beth Israel Deaconess Medical Center and Iowa University IRBs exempted informed consent for retrospective clinical data.GC, AB, DP, AJO-M, and AP-L conceived and designed the work, acquired the data, and analyzed and interpreted data. GC, AJO-M, and AP-L drafted the manuscript that was critically revised by the remaining authors for important intellectual content. AJO-M and AP-L had full access to all the data in the study and taken the responsibility for the integrity of the data and the accuracy of the data analysis. All authors approved the final version to be published and agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.GC was funded by the Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia through a PhD Scholarship (SFRH/BD/130210/2017). AB was supported by the NIH . AJO-M was funded by the FCT through a Junior Research and Career Development Award from the Harvard Medical School\u2014Portugal Program (HMSP-ICJ/0020/2011). GC and AJO-M were supported by grant PTDC/MED-NEU/31331/2017, and AJO-M by grant PTDC/MED-NEU/30302/2017, funded by national funds from FCT/MCTES and co-funded by FEDER, under the Partnership Agreement Lisboa 2020\u2014Programa Operacional Regional de Lisboa. AJO-M was also funded by a Starting Grant from the European Research Council under the European Union's Horizon 2020 research and innovation program (Grant Agreement No. 950357).The content of this study is solely the responsibility of the authors and does not necessarily represent the official views of the Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia, National Institutes of Health, European Research Council, and Harvard University or its affiliated academic healthcare centers.AJO-M was the national coordinator for Portugal of a non-interventional study to characterize a Treatment-Resistant Depression Cohort in Europe, sponsored by Janssen-Cilag, Ltd (2019\u20132020), is the recipient of a grant from Schuhfried GmBH for norming and validation of cognitive tests, and is the national coordinator for Portugal of trials of psilocybin therapy for treatment-resistant depression, sponsored by Compass Pathways, Ltd (EudraCT number 2017-003288-36), and of esketamine for treatment-resistant depression, sponsored by the Janssen-Cilag, Ltd (EudraCT NUMBER: 2019-002992-33). AP-L is a co-founder of Linus Health and TI Solutions AG; serves on the scientific advisory boards for Starlab Neuroscience, Magstim Inc., Radiant Hearts, and MedRhythms; and is listed as an inventor on several issued and pending patents on the real-time integration of non-invasive brain stimulation with electroencephalography and magnetic resonance imaging. None of the aforementioned agencies or companies had a role in the design and conduct of the study, in the collection, management, analysis, and interpretation of the data, in the preparation, review, or approval of the manuscript, nor in the decision to submit the manuscript for publication. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Ribosome profiling, or Ribo-seq, is the state-of-the-art method for quantifying protein synthesis in living cells. Computational analysis of Ribo-seq data remains challenging due to the complexity of the procedure, as well as variations introduced for specific organisms or specialized analyses.We present riboviz 2, an updated riboviz package, for the comprehensive transcript-centric analysis and visualization of Ribo-seq data. riboviz 2 includes an analysis workflow built on the Nextflow workflow management system for end-to-end processing of Ribo-seq data. riboviz 2 has been extensively tested on diverse species and library preparation strategies, including multiplexed samples. riboviz 2 is flexible and uses open, documented file formats, allowing users to integrate new analyses with the pipeline.github.com/riboviz/riboviz.riboviz 2 is freely available at Ribo-seq quantifies the \u2018translatome\u2019 of actively translated RNAs in cells to aid in the generation of this configuration file. The configuration file facilitates reproducible and transparent analyses, and allows the pipeline to run on various computing systems. riboviz 2 invokes both publicly available tools ; the Wellcome Trust ; the National Science Foundation ; the National Institutes of Health ; and start-up funds from the Human Genetics Institute of New Jersey at Rutgers University to P.S.Conflict of Interest: none declared.https://dx.doi.org/10.6084/m9.figshare.12624200. The development version of the source code is also available at https://github.com/riboviz/riboviz, where we welcome user comments and contributions.The source code and data underlying this article are available in figshare, at"} +{"text": "It can be argued, however, that the primary function of BMD measurements is to analyze the risk of (osteoporotic) fractures in an individual (Z-scores seem to be related to (stress) fracture risk, independent of sports discipline fracture risk. For early detection and prevention of LEA, research might need to focus more on the validation and use of objective blood markers.k et al. proposesdividual . In thisscipline . Furtherscipline ,5. Thesek et al. that higNo conflicts of interest, financial or otherwise, are declared by the authors. to the editor: The effect of physical activity on bone mineral density (BMD) is not uniform across different sports needs to be repudiated. There is a strong need to define activity-specific Z-scores.This will facilitate early identification of at-risk individuals and enable institution of corrective measures. As proposed by Jonvik et al. , this isNo conflicts of interest, financial or otherwise, are declared by the authors. to the editor: Jonvik et al. workout volume, intensity, density, and complexity and 2) recovery period strategies. Specifically in swimming, the voluminous general training period, the systematic exposure to the no-impact aquatic environment (training frequency is frequently greater than nine 2- to 3-h sessions/wk), and the very-high-intensity training sets of the specific and precompetition periods (k et al. discussed topics , highlig periods and in i periods , meaningNo conflicts of interest, financial or otherwise, are declared by the authors."} +{"text": "Excess deaths, including all-causes mortality, were confirmed for the first time in Japan in April 2021. However, little is known about the indirect effects of COVID-19 on the number of non-COVID-19-related deaths. We then estimated the excess deaths from non-COVID-19-related causes in Japan and its 47 prefectures from January 2020 through May 2021 by place of death. Vital statistical data on deaths were obtained from the Ministry of Health, Labour and Welfare. Using quasi-Poisson regression models, we estimated the expected weekly number of deaths due to all-causes excluding COVID-19 (non-COVID-19) and due to respiratory disease, circulatory disease, malignant neoplasms, and senility. Estimates were made separately for deaths in all locations, as well as for deaths in hospitals and clinics, in nursing homes and elderly care facilities, and at home. We defined a week with excess deaths as one in which the observed number of deaths exceeded the upper bound of the two-sided 95% prediction interval. Excess death was expressed as a range of differences between the observed and expected number of deaths and the 95% upper bound of the two-sided predictive interval. The excess percentage was calculated as the number of excess deaths divided by the expected number of deaths. At the national level, excess deaths from non-COVID-19-related all-causes were observed during April 19 to May 16, 2021. The largest excess percentage was 2.73\u20138.58% (excess deaths 689\u20132161) in the week of May 3\u20139. Similar trends were observed for all four cause categories. The cause-of-death categories which contributed to the excesses showed heterogeneity among prefectures. When stratified by place of death, excess mortality tended to be observed in nursing homes and elderly care facilities for all categories, in hospitals and clinics for circulatory disease, and at home for respiratory disease, malignant neoplasms, and senility. A caution is necessary that for the lastest three months (March\u2013May 2021), adjusted data were used to account for possible reporting delays. \u2022There are few studies on excess deaths due to non-COVID-19-related causes.\u2022Since April 2021, excess deaths from all causes other than COVID-19 have been observed in Japan.\u2022Excess mortality tended to be observed in nursing homes and elderly care facilities for all categories.\u2022These excesses have lasted for multiple, consecutive weeks, aligning with the fourth wave of COVID-19. Data from 2012 through May 2021 were employed in this analysis, which contained information on cause of death, place of residence (prefecture), and place of death for all people who have residency cards and died in Japan, regardless of nationality. We excluded those who died abroad, those who were staying in Japan for a short period of time (people without residency cards), as well as those whose place of residence or date of birth were unknown.2.2All-cause deaths except those due to COVID-19 ) and the following four causes were considered: respiratory disease , circulatory disease , malignant neoplasms (C00\u2013C97), and senility (R54). These cause groups and ICD-10 codes were selected based on previous analyses of comorbid conditions reported on death certificates where COVID-19 was listed as a cause of death as well 2.3To estimate the expected weekly number of deaths, we utilized the Farrington algorithm , which ib was modified to The Farrington algorithm computes a quasi-Poisson regression model, which is a generalized linear model accounting for overdispersion. It is designed to limit the data used for estimation: the expected number of deaths at a certain week us study . We cons studies . Since ilsewhere . It mustlsewhere , where tlsewhere . We spec2.4We defined a week with excess and exiguous deaths as one in which the observed number of deaths exceeded the upper bound or fell below the lower bound of the two-sided 95% prediction interval, respectively. Excess/exiguous deaths were expressed as a range that included the difference between the observed number of deaths and the expected number of deaths in addition to that between the observed number of deaths and the upper/lower bounds of the two-sided 95% prediction interval. The excess/exiguous percentage was also calculated as the number of excess/exiguous deaths divided by the expected number of deaths.3Between December 30, 2019, and May 30, 2021, 1990379 people in Japan died of non-COVID-19-related causes; no excess deaths from non-COVID-19-related all-causes were observed during consecutive weeks in 2020, but in 2021, excess deaths from non-COVID-19-related all-causes were observed between April 19 and May 16 . The lar4In 2020, exiguous deaths in hospitals and clinics and excess deaths at home were observed for malignant neoplasm-related deaths starting in April, which coincided with the first declaration of a state of emergency against COVID-19 in Japan. At that time, it was necessary to concentrate limited medical resources on patients with severe COVID-19 and potentially infected patients, and patients receiving long-term inpatient care were encouraged to be transferred to nursing homes, elderly care facilities, or home care . There aIn 2021, excess deaths from non-COVID-19-related all-causes were observed during the fourth wave of COVID-19 that began in April. Excess deaths were observed simultaneously for all major causes of deaths in Japan. As opposed to the long-term health effects of restrictions on access to non-urgent medical care and changes in lifestyle, it is possible that the strain on healthcare systems caused by the rapid increase in COVID-19 patients has affected acute and end-of-life care . In partThe limitations of this study lie in its reliance on provisional data in the latest 3 months, March\u2013May 2021, and in the assumptions applied to the model. In addition, the Farrington algorithm for estimating excess/exiguous deaths only evaluates whether there are more or fewer deaths than in previous years; therefore, we did not directly assess causality between the COVID-19 pandemic and the number of deaths in any given category. In other words, the fact that excess deaths were observed does not allow us to conclude that they were the result of indirect effects of the pandemic. It is possible that our estimates also include the effects of unobserved contingencies unrelated to the pandemic.It is important to note that due to the nature of the methodology used in this study, data from 2020 was used to estimate 2021. In some locations and at some times, exiguous deaths were observed during 2020, which may have exerted an effect on the expected number of deaths around the same time in 2021. However, our estimates in 2021 depend not only on data from 2020, but also on data from before 2020 (2016\u20132019), and in this sense 2020 alone is unlikely to have a significant impact on current estimates of excess/exiguous deaths. The alternative, namely excluding 2020 data when calculating 2021 estimates, would jeopardize the estimation of longitudinal time trends and seasonality, which we believe would constitute an inappropriate mathematical approach to estimating excess deaths during the pandemic.5Using vital statistics data on deaths from January 2012 to May 2021 obtained from MHLW, we estimated excess deaths from all causes other than COVID-19 , as well as deaths from respiratory disease, circulatory disease, malignant neoplasms, and senility, on a weekly basis during 2020\u20132021 compared to previous years. We found that since April 2021, excess deaths lasting multiple, consecutive weeks have been observed for the first time in Japan for all cause categories, coinciding with the onset of the fourth wave of COVID-19. In addition, when stratified by place of death, excess deaths tended to occur in nursing homes and elderly care facilities for all categories, in hospitals and clinics for circulatory disease, and at home for respiratory disease, malignant neoplasms, and senility. These findings may indicate that COVID-19-associated healthcare system strain has had profound consequences for acute and end-of-life care. A caution is necessary that for the lastest three months (March\u2013May 2021), adjusted data were used to account for possible reporting delays.Ethical approval was granted by the ethics committee of the National Institute of Infectious Diseases, under authorization number 1174.10.13039/501100003478Ministry of Health, Labour and Welfare of Japan and the 10.13039/501100001700Ministry of Education, Culture, Sports, Science and Technology of Japan (21H03203), and by 10.13039/501100009023Precursory Research for Embryonic Science and Technology from the 10.13039/501100002241Japan Science and Technology Agency (JPMJPR21RC). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.The present work was supported in part by grants from the Not applicable.The mortality data have been obtained through a restricted data-use agreement with the Ministry of Health, Labour and Welfare, Japan, and are therefore not available for public dissemination.Not applicable.Shuhei Nomura: Conceptualization, Data curation, Formal analysis, Funding acquisition, Investigation, Methodology, Project administration, Resources, Validation, Writing \u2013 original draft, Writing \u2013 review & editing. Akifumi Eguchi: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Validation, Writing \u2013 original draft, Writing \u2013 review & editing. Cyrus Ghaznavi: Investigation, Writing \u2013 original draft, Writing \u2013 review & editing. Yuta Tanoue: Data curation, Formal analysis, Investigation, Methodology, Validation, Writing \u2013 review & editing. Takayuki Kawashima: Data curation, Formal analysis, Investigation, Methodology, Validation, Writing \u2013 review & editing. Daisuke Yoneoka: Data curation, Formal analysis, Funding acquisition, Investigation, Methodology, Validation, Writing \u2013 review & editing. Lisa Yamasaki: Investigation, Writing \u2013 review & editing. Motoi Suzuki: Data curation, Investigation, Project administration, Resources, Writing \u2013 review & editing. Masahiro Hashizume: Conceptualization, Data curation, Funding acquisition, Investigation, Project administration, Resources, Supervision, Writing \u2013 review & editing.The authors declare that they have no competing interests."} +{"text": "Sex and gender are concepts that are often misunderstood and misused, being utilized in a biased, preconceived, interchangeable way. Sex and gender medicine is generally overlooked, despite the profound impact of sex and gender on health outcomes. The aims of the present rapid systematic literature review were (i) to assess the extent to which sex- and gender-sensitive topics are covered in medical courses; (ii) to assess the need for and willingness toward integrating/incorporating sex and gender medicine into health-related education; (iii) to identify barriers and facilitators of the process of implementation of sex and gender medicine in medical teaching, mentoring, and training; and (iv) to evaluate the effectiveness of interventional projects targeting curriculum building and improvement for future gender-sensitive physicians. Seven themes were identified by means of a thematic analysis, namely, (i) how much sex- and gender-based medicine is covered by medical courses and integrated into current medical curricula, (ii) the knowledge of sex and gender medicine among medical and allied health profession students, (iii) the need for and willingness toward acquiring sex- and gender-sensitive skills, (iv) how to integrate sex- and gender-based medicine into medical curricula in terms of barriers and facilitators, (v) existing platforms and tools to share knowledge related to sex and gender medicine, (vi) sex- and gender-based medicine aspects in the post-medical education, and (vii) the impact of sex- and gender-sensitive topics integrated into medical curricula. Based on the identified gaps in knowledge, further high-quality, randomized trials with larger samples are urgently warranted to fill these gaps in the field of implementation of gender medicine in educating and training future gender-sensitive physicians. Despite sounding like they overlap, they are two separate concepts. \u201cSex\u201d is, indeed, a biological variable linked to the genetic/genomic and post-genetic/post-genomic, endocrinological , and phenotypic/anatomic/phenomic (reproductive organs) components. In contrast, \u201cgender,\u201d which is a subjective variable (self-identification/self-declaration), relates to one\u2019s personal, as well as societal, cultural, and political experiences. Both sex and gender variables and their subtle interactions can impact health-related outcomes . The still-ongoing \u201cCoronavirus Disease 2019\u201d (COVID-19) pandemic showcases how profoundly clinical (diagnostic and prognostic) outcomes can be affected by sex and gender and how it is of crucial importance to consider such parameters .Several healthcare organizations and institutions, like the USA \u201cNational Academy of Medicine,\u201d previously known as the \u201cInstitute of Medicine\u201d (IoM), and the Canadian Institutes of Health, have underlined how these variables are, indeed, not restrained to reproductive and sexual health and wellbeing only, but they concern the entire human organism as a whole, from a biological, as well as from psychological and societal points of view. Furthermore, devising strategies and conceptual frameworks that integrate sex- and gender-related aspects would influence the way healthcare provisions are delivered and, generally speaking, all the biomedical practices. This would pave the way for the achievement and implementation of the so-called personalized/individualized medicine, where the precise needs of each individual are taken into account, instead of a \u201cone-size-fits-all\u201d approach.Sex- and gender-specific differences can, indeed, dramatically impact the natural story of a disease, from its etiopathogenesis to the response to treatment .Despite the importance of this topic, sex- and gender-specific medicine is still overlooked in the academic arena , with inConcerning COVID-19, research has found that, despite the implications of sex and gender for COVID-19 diagnosis, treatment, and prognosis, these variables are rarely accounted for. For instance, out of 75 COVID-19 vaccines trials, only 24% explicitly presented endpoint outcomes stratified by sex and only 13% of them discussed sex-specific differences [All this shows the importance of integrating/incorporating sex- and gender-based medicine into clinical teaching. Sex- and gender-based medicine has been classically defined as \u201ca study of the differences in men\u2019s and women\u2019s normal function and in their experience of the same diseases,\u201d according to Marianne Legato .The aims of the present rapid systematic literature review were (i) to assess the extent to which sex- and gender-sensitive topics are covered in medical courses; (ii) to assess the need for and willingness toward integrating/incorporating sex and gender medicine into health-related education; (iii) to identify barriers and facilitators of the process of implementation of sex and gender medicine in medical teaching, mentoring, and training; and (iv) to evaluate the effectiveness of interventional projects targeting curriculum building and improvement for future gender-sensitive physicians.The review study protocol was a priori devised according to the \u201cPreferred Reporting Items for Systematic Reviews and Meta-Analyses-Protocol\u201d (PRISMA-P) 2015 guidelines . The proA rapid systematic review of the literature was condOne database was deemed enough since a rapid systematic review of the literature generally includes a streamlined methodology with respect to classic approaches deployed in conducting systematic reviews of the literature (including mining only one database).Inclusion and exclusion criteria were devised according to the population/intervention (exposure)/comparator (comparison)/outcome/study design (PICOS) criteria. Inclusion criteria were original studies of any type in which either qualitative/quantitative or mixed methods research had been conducted (S), surveying medical or allied health profession students , as well as professors from medical faculties (P).Articles were retained if performing any comparison, including attendance of sex- and gender-sensitive seminars, lectures, or workshops versus nonattendance; assessing the best form of delivering gender-sensitive topics , among others).Articles dedicated to the implementation of \u201clesbian, gay, bisexual, transexual/transgender\u201d (LGBT)/\u201clesbian, gay, bisexual, transexual/transgender/queer/intersex\u201d (LGBTQI)-related healthcare were excluded from the present review. Studies assessing the exposure of students to men\u2019s health, women\u2019s health, gender diversity, gender inclusion, and related topics were not retained in the present review. Articles surveying students and/or professors from non-medical faculties were excluded as well.Further details are reported in Biology of Sex Differences, Health Care for Women International, Journal of Women\u2019s Health (Larchmt), GMS Journal of Medical Education, BMC Medical Education, Gender Medicine, and Education for Health (Abingdon), were hand-searched.Extensive cross-referencing was applied, manually screening the reference list of each potentially eligible article. Target journals, including Relevant data were abstracted by two authors independently from retained studies and were synthesized using a thematic approach . In the Critical quality appraisal and risk of bias were not performed since they are streamlined steps within the rapid systematic review literature methodology .The findings of the present rapid systematic review are reported according to the \u201cPreferred Reporting Items for Systematic Reviews and Meta-Analyses\u201d (PRISMA) 2020 guidelines .The initial literature search yielded a pool of 8464 items. After removing duplicates and discarding irrelevant articles, twenty-five studies ,36,37,38Sex- and gender-based medicine is rarely and inconsistently covered by medical courses and is not organically/coherently integrated/combined into medical curricula. Thande et al. assessedMiller et al. carried In another study, Miller et al. obtainedHenrich and Viscoli carried In another study, Henrich et al. surveyedFinally, Nocon et al. surveyedn = 4 from the second year) correctly stated that clinical trials should provide outcomes stratified by gender.In the study by Miller et al. , the medp = 0.04) and working in a private entity was associated with applying the principles of sex- and gender-based medicine in the daily practice of physiotherapy. Other statistically significant predictors were knowledge/understanding of sex- and gender-sensitive topics, their perceived importance and impact on human health and disease, and need/willingness to attend courses/lectures/workshops or clerkships related to sex- and gender-based medicine.Bisconti et al. evaluaten = 536) of instructors, lecturers, and educators about the teaching and testing content. The respondents felt that communicative and social competencies and skills were of crucial importance in the field of medical education, teaching, and training.Exenberger et al. evaluateSteinb\u00f6ck et al. describen = 50). Applicants were willing to acquire skills to become gender-sensitive doctors, strongly advocating for inclusive healthcare and equity in access to health provisions. According to the respondents, the best path toward fully incorporating sex and gender medicine into daily clinical practice should start from acquiring basic biomedical knowledge about sex- and gender-specific differences and their impact on human health and disease, gradually developing social and communicative skills, and competence. The deployment of audiovisual interactive materials and simulated practical situations could be useful within this strategy. Finally, students emphasized the importance of having gender-sensitive mentors and educators during medical studies.Scholte et al. conducteGaida et al. describeJenkins et al. surveyedvan Leerdam et al. performeFinally, B\u00f6ckers et al. describeVerdonk et al. qualitatPark et al. developeLudwig et al. devised The 10-step approach implemented by Ludwig et al. was simiMoreover, Chin et al. and McGrIn a study by Nachtschatt et al. , it was Finally, building on previous research and proposals, Tannenbaum and Moineau suggesteAuditing and mapping extant sex- and gender-sensitive topics is a good practice, as indicated by Song et al. . Existinhttp://egender.charite.de, accessed on 2 February 2022. The platform was devised with the aim of providing future health professionals with adequate knowledge and communication competence and skills on sex- and gender-specific differences and their impact on human health and disease. From a pedagogic and educational perspective, the tool can support a blended learning teaching framework and is based on the concept of constructivism. The content is of high quality and has been specifically developed by experts from seven universities: it is both of broad scope and nature and specific to each medical specialization. Modules consist of textual, audio-visual, and interactive material, and the number of registered users has been gradually increasing, showing more interest in sex and gender medicine.For instance, Seeland et al. describewww.gendermed-wiki.de, accessed on 2 February 2022) is an online exchange platform funded by the German Federal Ministry of Education and Research [\u201cGenderMed-Wiki\u201d is developed by the Institute of Gender in Medicine (GiM), based at the Charite University Hospital, Berlin, Germany. The FUTURE platform includes the open knowledge database developed by Stanford University and adjusted for the Swedish audience [The GenderMed database (ry 2022) .Another existing tool is the Canadian \u201cGender Lens Tool.\u201d Weyers et al. translathttp://www.laurabushinstitute.org/cme/default.aspx, accessed on 2 February 2022).Finally, the Online Continuing Medical Education and Certificate Program in Sex- and Gender-Specific Health, under development with the support of the Laura W. Bush Institute for Women\u2019s Health, is a continuing professional development site or mainstream (n = 72) gender medicine program. A third cohort of general practitioner trainees (n = 60) acted as the control cohort. Participants were administered the \u201cNijmegen Gender Awareness in Medicine Scale\u201d and a 16-item questionnaire concerning gender knowledge. The modular approach toward teaching gender medicine was found to statistically significantly improve gender knowledge (p = 0.049), whereas the three cohorts did not differ in terms of gender sensitivity or gender role ideology. Interestingly, female GP trainees were more gender-aware than their male counterparts.Finally, Dielissen et al. evaluateIntegrating sex- and gender-sensitive topics into medical curricula significantly improved students\u2019 knowledge of sex and gender medicine. Siller et al. surveyedIn a study by Chin et al. , the outIn a study by Park et al. , particiAddressing healthcare disparities and inequities and ensuring equitable and customized access to healthcare provisions are a societal onus and a crucial step for implementing initiatives and programs based on precision and individualized/personalized/stratified medicine. Sex and gender medicine should be a fundamental component of medical curricula, being a major asset of precision medicine. Incorporating and integrating sex- and gender-related variables into clinical research and practice would significantly enhance and deepen our understanding of the subtle, complex, and non-linear impact of sex and gender on human health and disease.However, findings from the present rapid systematic review of the literature show that the majority of medical students, as well as clinical residents and fellows, have never had or rarely had the opportunity of discussing the topic of sex and gender medicine with their instructors, even though they perceive the importance of such topics. Current medical curricula, both at the graduate and post-graduate levels, suffer from profound gaps with regard to the implementation of teaching components related to sex and gender medicine.Moreover, there are barriers to the full integration of sex- and gender-specific knowledge into the biomedical syllabus, which include preconceived, biased opinions and ideas about sex and gender, the lack of adequate learning material and systematic teacher training, and an innovative strategy based on social and communicative competence and skills, as well as a perceived minor impact, translational added value, or even irrelevance of sex and gender medicine on daily clinical practice . FurtherBased on the findings of the articles overviewed in the present review, we can identify the main desired features of a sex and gender medical course. This should be longitudinally integrated with other courses, with a strong clinical focus, covering multiple competency levels and making use of web/interactive resources and tools, with measurable learning objectives and outcomes and concrete, content-oriented, practical goals. These and other desired features of a sex- and gender-medicine course are overviewed in Equipping future physicians with a clear understanding of the influence and effects of sex and gender on health is paramount in advancing the achievements of patient-centered care.It is, therefore, crucial to change the attitudes of medical students toward sex-/gender-sensitive medicine.To the best of our knowledge, this is the first systematic review on the topic of integrating sex and gender medicine into medical curricula. Our review significantly complements and adds to the narrative reviews by Ruiz-Cantero et al. and by LCOVID-19 as a showcase of the so-called \u201cgendered diseases\u201d has highlighted the importance of integrating and incorporating sex- and gender-sensitive topics into the biomedical curriculum. The present rapid systematic review of the literature identified seven main educational themes, including the major barriers and facilitators of the implementation of sex and gender medicine in the educational training of medical students, residents, and fellows . However"} +{"text": "The National Mental Health Survey of India 2015-16 indicated a large treatment gap of 70-92% for mental disorders and a paucity of mental health specialists in the country. In order to address this treatment gap and develop human resources, the National Institute of Mental Health and Neuro Sciences (NIMHANS), Bengaluru, India, with impetus from the Ministry of Health and Family Welfare, Govt. of India, launched the online course of Diploma in Community Mental Health for Psychologists.The course was designed with the objective of training individuals with a Master\u2019s Degree in Psychology, in providing first-level psychological care in the community.The course is a 3-month online programme comprising of approximately 25 hours of self-paced e-learning and 11 hours of live real-time interactive discussion via video conference. The course comprises of 6 modules, with an assessment at the completion of each module. Pre- and Post-Assessment is conducted to evaluate competencies achieved.On successful completion of the course, trainees are expected to have achieved competencies to: Screen for and identify mental health problems in adults and children, and understand factors influencing them; Understand management options; Conduct interview-based functional developmental assessment for intellectual deficits; Conduct first-level brief psychosocial interventions; Make appropriate referrals to Mental Health Professionals and other health professionals.This digitally-driven online course is a viable option for development of human resources on a large scale, in a resource-scarce country such as India.No significant relationships."} +{"text": "This research aimed to explore the interaction between lotus root polysaccharides (LRPs) and phenolic compounds, and to study the effects of phenolic binding on the structural and functional properties of LRPs. The influences of pH, temperature, and NaCl and phenol concentration on the binding ratio of gallic acid (GA)/epigallocatechin (EGC) to LRPs were evaluated. LRP-GA/EGC complexes with different phenolic binding amounts were then prepared and characterized via ultraviolet\u2013visible (UV\u2013Vis) and Fourier-transform infrared (FTIR) spectroscopy, and average molecular weight (MW) measurements. The results suggest that hydrogen bonds contributed to the binding of GA/EGC and LRPs. The phenolic binding led to significant changes in the structure and MW of LRPs. Moreover, antioxidant activity and the macrophage-stimulating effect of LRPs were improved after binding with GA/EGC, depending on the binding amount and type of polyphenol. Interestingly, LRP-GA/EGC complexes with polyphenol binding amounts of 105.4 mg/g and 50.71 mg/g, respectively, showed better stimulation effects on the anti-inflammatory cytokine IL10 secretion of macrophages when compared to LRPs. These results show the great potential of phenolic binding to be applied to improve the structure and functional activity of LRPs. Nelumbo nucifera Gaertn.) root is one of the most popular aquatic vegetables cultivated and consumed in Southeast Asia. It has attracted considerable attention due to its edible properties and therapeutic potential . B. B13]. Bmixtures B, and itFTIR spectrum analysis was carried out to characterize the binding of GA and EGC on LRPs. 67 cm\u22121) ,30,31. T LRP-GA3 . The O-HThe FTIR spectrum of EGC, physical mixture of LRPs and EGC, and LRP-EGC complexes are shown in ic rings ,33,34. Pbrations ,36. Aftevenumber . The absThe elution profiles of LRPs and LRP-phenol complexes as a function of the elution time obtained by the HPSEC-MALL-RI method are presented in studies ,16 had sAs shown in ectively E. The hiThe antioxidant activities of LRPs, LRP-GA/EGC complexes, and their mixtures evaluated by FRAP are presented in the LRPs . The FRANO is an important immune-regulatory signaling molecule whose release from macrophages can suppress the growth of bacterial infection or cancer cells . FurtherAs mentioned above, LRP-GAectively C, but thIn this work, the binding interactions between GA/EGC and LRPs were studied by evaluating the effect of environmental factors on the binding amount of GA/EGC to LRPs, and measuring the UV\u2013Vis spectra, FTIR spectra, and MW of LRPs-GA/EGC complexes. It was demonstrated that GA/EGCs successfully bond to LRPs by hydrogen bond interactions, leading to the changes in structure and MW distribution of LRPs. The binding of phenolic compounds significantly increased the DPPH radical scavenging activity and FRAP capacity of LRPs, and the increase degree was dependent not only on the phenol type but also on the mass ratio of the phenols in complexes. Moreover, GA and EGC combined to LRPs promoted the NO production of macrophages in normal conditions; EGC with a mass ratio of 698.71 mg/g bound to LRPs showed inhibitory effects on the NO over-production stimulated by LPS. Binding with GA/EGC did not enhance the TNF-"} +{"text": "In order to explore and further understand the efficacy of donepezil (DNP) in the treatment of Alzheimer's disease (AD), this research was conducted based on network pharmacology and molecular docking.Compounds of DNP and its effective targets were collected using the TCMSP Chinese medicine system pharmacology database. Disease targets were screened and selected utilizing GeneCards, TTD, DrugBank, CTD, and other online databases. Then, Venn diagrams were generated to identify the intersections. A diseases-drug-active ingredient-key target protein interaction (PPI) network was constructed using the STING database. GO and KEGG enrichment analyses were conducted to predict the function and mechanism of DNP, which were visualized by graphs and bubble charts. After the screening, the top five interacting targets in the PPI network and the compound containing the most active target were selected for molecular docking.The study received 110 potential targeting genes and 155 signaling pathways. A strong association between DNP and modulation of chemical synaptic transmission and the regulation of trans-synaptic signaling is noted. Signaling pathways related to the proliferation, differentiation, and survival of cells are also found positively relative. The results revealed that the mechanism of its therapeutic effect is multi-component, multi-target, and multi-pathway, laying a foundation for the follow-up in-depth study of the mechanism of DNP in the treatment of AD.This research provides a superior prediction that AD could be treated using DNP which targets the key proteins and essential pathways associated with the recovery of AD. Alzheimer's disease (AD) is a neurodegenerative disorder in the central nervous system, prevalently observed among the elderly and near-elderly, characterized by progressive cognitive impairment, and memory degeneration . Despite the effectiveness of the existing therapies in treating the symptoms of AD, no significant influence has been found in curing or preventing the disease stimulations . With the structure acquired, data of the donepezil targeted sites were collected from four separate databases: swisstarget (http://www.swisstargetprediction.ch/), pharmmapper (http://lilab-ecust.cn/pharmmapper/), batman-tcm (http://bionet.ncpsb.org.cn/batman-tcm/), and stitch (http://stitch.embl.de/), each with a different standard for data selection. For swisstarget, data with Probability*>0 was included. For batman-tcm, the inclusion standard was set Score cutoff\u2265 20. No standard was set for screening in stitch or pharmmapper. The corresponding genes of the donepezil target sites were collected through Uniprot (http://www.uniprot.org/www.uniprot.org/), utilizing data downloaded in pharmmapper.First, the structural formula of donepezil was downloaded from PubChem from three aspects, Biological Process (BP), Cellular Component (CC), and Molecular Function (MF), with p-value cutoff and q-value cutoff both as 0.05. The results from each aspect were visualized with a graph and bubble chart.With the selected genes, KEGG pathway enrichment analysis was conducted to predict its mechanism of action using DOSE, clusterProfiler, and pathview packages (Bioconductor), with p-value cutoff and q-value cutoff both as 0.05. Columns and bubble charts were generated to visualize the result.*>0, score cutoff\u226520 was screened and downloaded from the swisstarget and batman-tcm databases. The structure was also searched in the stitch and pharmmapper databases to further verify the results. The structure was visualized using PubChem was utilized to visualize the intersection result , including 10 BP, 10CC, and 10 MF. Therefore, it can be concluded that the mechanism of the therapeutic effect of donepezill to AD could be associated with biological processes such as positive regulation of RNA polymerase II promoter, signal transduction, and positive regulation of DNA template transcription. At the same time, a variety of substances such as plasma membrane, cytosol, and extracellular space are involved, such as protein binding, enzyme binding, transcription factor activity, sequence-specific DNA binding, same protein binding, and other molecular functions.Genes for potential target sites were selected using the R package from \u201corg.Hs.eg.db\u201d database, and GO enrichment analysis was conducted using the DOSE, clusterProfiler, and pathview packages (Bioconductor) from three aspects, Biological Process (BP), Cellular Component (CC), and Molecular Function (MF), with p-value cutoff and q-value cutoff both as 0.05. The results from each aspect were visualized with a bubble and bar chart . 30 GO lP < 0.05, FDR < 0.05), and the top 20 are listed by Count number (In KEGG pathway enrichment analysis, 155 signaling pathways are screened (t number . The KEGt number .The development of drugs with clinical value has met with extreme obstacles (Becker et al., DNP is a kind of AChEI, which functions as an inhibitor to the cholinesterase enzyme and sustains the activity of acetylcholine at cholinergic synapses. DNP has been shown to significantly improve cognition and daily function and some behavior manifestations of patients with AD (Massoud and Gauthier, The study received 110 potential targeting genes and 155 signaling pathways in total. The diversity of the associated genes and pathways demonstrate that the mechanism of DNP in the treatment of AD is multifactorial. A strong association between DNP and modulation of chemical synaptic transmission and the regulation of trans-synaptic signaling is noted, further confirming the ability of DNP in sustaining the activity of synapses. In addition, signaling pathway related to the proliferation, differentiation, and survival of cells are also found positively relative, such as Ras and MAPK signaling pathways, suggesting a potential of promoting the growth and proliferation of neurons using DNP.The cholinergic system plays an indispensable role in neuronal function in memory, learning, and other essential aspects of cognition and plays a wider role in the promotion of neuronal plasticity (Drachman and Leavitt, in vivo or in vitro, and deficiencies remain noticeable. However, this study can provide new ideas and directions for further exploration of relevant experiments.In this study, due to the incomplete information in the database and unclear interaction between DNP and other factors, the prediction results have certain limitations. This study did not verify the possible molecular mechanism of DNP in the treatment of AD syndromes either The simulation experiment data used to support the findings of this study are available from the corresponding author upon request.LL and PF prepared the figures. JY and YZ finished the paper. All authors contributed to the article and approved the submitted version.This work was supported in part by the Science and Technology Commission of Shanghai Municipality, General Program, No.20ZR1456400.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "K-means with CNN are the six modules examined and evaluated. Experiments are carried out on various types of images such as Magnetic Resonance Imaging (MRI) for brain data analysis, dermoscopic for skin, microscopic for blood leukemia, and computed tomography (CT) scan images for lungs. After combining all of the datasets, we have constructed five subsets of data, each of which had a different number of images: 50, 100, 500, 1000, and 2000. Each of the models was executed and trained on the selected subset of the datasets. From the experimental analysis, it is observed that the performance of K-means with CNN is better than others and achieved 96.45% segmentation accuracy with an average time of 9.09\u2009seconds.Recent improvements in current technology have had a significant impact on a wide range of image processing applications, including medical imaging. Classification, detection, and segmentation are all important aspects of medical imaging technology. An enormous need exists for the segmentation of diagnostic images, which can be applied to a wide variety of medical research applications. It is important to develop an effective segmentation technique based on deep learning algorithms for optimal identification of regions of interest and rapid segmentation. To cover this gap, a pipeline for image segmentation using traditional Convolutional Neural Network (CNN) as well as introduced Swarm Intelligence (SI) for optimal identification of the desired area has been proposed. Fuzzy C-means (FCM), K-means, and improvisation of FCM with Particle Swarm Optimization (PSO), improvisation of K-means with PSO, improvisation of FCM with CNN, and improvisation of To advance the efficiency and accuracy of the medical diagnostic system, especially those that are distributed in complex areas , several live line diagnostic models . Many rUsed traditional segmentation techniques are the type of unsupervised machine learning and useful to find out groups or different patterns in medical data. In general terms, it's an unsupervised activity that divides unstructured data into several groups based on their similarity and dissimilarity . The moThe number of research articles available was large but lack of appropriate comparisons of Traditional medical image Segmentation, SI, and CNN-based segmentation.Medical data or image segmentation is a challenging task and still lots of improvements are needed to develop a better diagnosis system.Existing CNN-based models need a lengthy system-training period.Suffering from the over-fitting problems and need to solve such kind of problem regarding the medical diagnosis system that helps to detect the diseases in early stage. The overfitting problem in deep learning usually occurs when the image count is small in the target.The existing system had to be developed and updated in real-time scenarios.There are no studies that have established a single standard segmentation model for distinct picture types from various organs.Unsupervised clustering-based medical image segmentation is a default method that aims to collect a set of objects or pixels into subsets or collections by the background and front of the image. The goal is to create clusters or parts that fit inside but are very different from each other. In simple terms, pixels in the same category should be as similar as possible, and objects in the same category should be very different from those in another cluster. Some challenging factors given that gave us the motivation are as follows:To study the existing medical image segmentation approaches with different algorithms.Develop a novel pre-processing for medical images like image quality enactment, hair removal from dermoscopic images, and blast nucleus improvement for microscopic images.K-means are used as unsupervised machine learning approaches with PSO as swarm intelligence and CNN as a deep learning mechanism. A novel fitness function is presented here that replaces pixels to increase segmentation quality.To segment medical images, FCM and To validate the proposed framework, performance parameters such as Precision, Recall, F-measure, Accuracy, Error, Matthews's Correlation Coefficient (MCC), Dice coefficient (DC), Jaccard Coefficient (JC), and time being calculated and related with existing works.Nowadays, medical image segmentation using clustering is a basic requirement for lots of purposes like abnormal region detection, automatic extraction, data organization, etc. In the segmentation of medical data, high-quality clustering techniques are critical. Thus, in this research, we proposed a comparative framework for medical image segmentation with SI as well as CNN technique and the main contributions are as follows:This research article deals with a comparative study for medical image segmentation and the rest of the article is systematized into different sections. Segmentation is widely used in various sectors such as split geographical regions, fruit from trees, flood for damage reports, recognition of traffic signs, and road collapses. Chouhan et al. surveyedHowever, image segmentation is extensively used in human disease detection and diagnosis. For precise detection of the disease, initially, it requires identifying the region of interest from the captured images. In this study, a comprehensive description of the most important state-of-the-art medical image segmentation techniques is given. Here, we consider a mixed survey of segmentation for different types of medical data.K-means and concluded that using PSO as swarm intelligence is a useful step. The results of the experiments showed that PSO-based segmentation is more accurate than FCM, Adaptive Regularized Kernel FCM, and K-means In one of our previous studies, we have developed an IoT-based data collection system for skin lesions where we classified various skin lesions using deep learning-based ensemble algorithms . For skiK-means-based clustering is studied. The proposed scheme was compared to a previous clustering method, and the findings showed that the system's performance was better in terms of efficiency, computational burden, and quality attributes esBased on the above-written hybrid segmentation algorithm using FCM with PSO, we obtained better-segmented results as compared to the only FCM as well as K-means also and results with the original medical image shown in K-means clustering algorithm used as a medical image hybrid segmentation and the algorithm of K-means with PSO segmentation is written below in The concept of PSO along with the Based on the above-written hybrid segmentation algorithm using K-means with PSO, we obtained better-segmented results as compared to the FCM, K-means, and FCM with PSO also and results with the original medical image shown in This is the third module of implementation where we used again two different scenarios that are described below.In this scenario, we utilize the concept of CNN as a deep learning mechanism along with the FCM as a medical image hybrid segmentation. This hybrid method is currently used in most of the existing medical image segmentation research. First, we train the model using lots of already segmented images in terms of background and foreground images having 3 dimensions (RGB). Usually, segmented medical images, which are fed into the neural network, are reduced in data dimensions, reduce the system processing time as well as complexity and help to reduce the over-fitting problems and hybrid CNN mechanism shown in K-means, and improvisation in FCM with PSO, improvisation in K-means with PSO and results with the original medical image shown in We obtained better-segmented results for the proposed hybrid mechanism of FCM with CNN as compared to the FCM, K-means used in this scenario and similar to K-means is shown in The concept of CNN as a deep learning mechanism along with the K-means with PSO segmentation written below in K-means, FCM with PSO, CNN, and K-means with PSO, we achieved better-segmented results for the proposed hybrid mechanism of K-means with CNN, and results with the original medical image are displayed in In comparison to FCM, F-measure, Accuracy, Error, MCC, DC, JC, and time.Finally, performance parameters for different types of datasets are calculated and compared using a comparison framework simulation in terms of Precision, Recall, https://www.med.upenn.edu/sbia/brats2018/data.html\u201d having MRI images [The sample images of the BraTS dataset are shown in I images . For thehttps://homes.di.unimi.it/scotti/all/\u201d and the sample of ALL-IDB dataset images is shown in The dataset contains 2008 images that were collected in September 2005 in the Image Processing Department of Computer Science-Universit\u00e0degliStudi di Milano\u201d . The ALLThe used dataset contains approximately 39,000 blood counts, and oncologists labeled the lymphocytes. We resize the original microscopic images of blood samples into a size of 256\u2009\u00d7\u2009256 and a total of 2000 images were used in this research work.https://challenge2018.isic-archive.com/task1/training/ [It contains the human lesion analysis toward melanoma detection and the dataset is in the form of dermoscopic images. The dataset is available from raining/ . To capthttp://www.via.cornell.edu/lungdb.html\u201d [The database currently consists of an image set of 50 lung CT scans for research purposes which is publicly available from \u201cdb.html\u201d . A samplAfter executing the methodology's outlined steps, the performance has been evaluated in terms of several parameters, as discussed in the result and discussion section.F1 score, to assess segmentation quality. In order to analyze and comprehend the ability of the models, we have made use of each of these measures.In this part, we have outlined the assessment measures used to verify the effectiveness of the suggested techniques. First of all, we have observed quantitative metrics such as Accuracy. In most cases, the efficiency of a models is measured in terms of its accuracy. However, in medical image segmentation, the model's accuracy is insufficient to provide a precise understanding of the model. Therefore, there are several additional measures, such as precision, recall, Error, and Moreover, we have considered similarity metrics such as Matthews's Correlation Coefficient (MCC), Dice coefficient (DC), and Jaccard Coefficient (JC). Each similar metric has a few special characteristics to evaluate the true performance of the selected segmentation techniques. If all of the probabilistic methods, including true positives, true negatives, false negatives, and false positives, provide a high score, then the MCC algorithm will generate a higher score . SimilarK-means, and improvisation of FCM using PSO, improvisation of K-means with PSO, improvisation of FCM with CNN and improvisation of K-means with CNN. Simulation results of the offered scenario are shown in In this research work, we proposed a comparative framework for the medical image segmentation from various types of images such as MRI, Dermoscopic, Microscopic, and CT-scan images using the six different scenarios such as FCM, There are five separate sets of data that included a varied number of images considering 50, 100, 500, 1000, and 2000 images. We have considered an equal number of images from each dataset to produce five specified subsets. Then we applied various segmentation techniques to the subsets of data. The concluded result has been described using different metrics such as precision, recall, accuracy, and F-measure, as shown in K-means, FCM with PSO, K-means with PSO, FCM with CNN, and K-means with CNN, respectively. So, we can say that the effect of CNN on K-means for medical image segmentation is far better than other combinations. However, we need to validate the model based on similar parameters such as MCC, DC, JC, and computational time.From K-means with CNN is superior to other modules for all the similarities metrics such as MCC, JC, and CD. The required time for K-means with CNN is slightly higher than other models. However, the change is extremely minute and may safely be ignored as a result. FCM with CNN is the second most successful segmentation technique based on both quantitative and similarity metrics. Also, it is transparent that the CNN-based optimized segmentation techniques performed better than both swarm intelligence and traditional methods.Therefore, the simulation results based on the similar values have been given in Achieving maximum accuracy is the goal of the proposed framework with a fast response to segmentation with pre-processing. The proposed framework offers an extremely self-configurable and standalone mechanism with lots of deep learning interfaces. In addition, our suggested framework generated robust models for segmenting the region of interest in various issues. So, it is a universal framework for medical image segmentation. However, to validate the efficiency of the system, we need to compare it with state-of-the-artwork based on their accuracy in K-means with CNN.It is clear that the previous study was conducted on a single issue such as skin lesions, brain tumors, lung cancer, or leukemia (as shown in K-means, improvisation of FCM using PSO, improvisation of K-means with PSO, improvisation of FCM with CNN, and improvisation of K-means with CNN. We proved the functionality of K-means with CNN is a powerful hybrid mechanism that achieves 96.45% accuracy whereas, other mechanisms achieve 85.72%, 86.61%, 87.54%, 88.64%, and 92.21% FCM, K-means, and improvisation of FCM using PSO, improvisation of K-means with PSO, improvisation of FCM with CNN and improvisation of K-means with CNN respectively. We also found that CNN-based optimized algorithms performed well compared to optimized swarm intelligence and/or traditional methods. We expect that in the future, it will aid in the transition of medical image segmentation from research laboratories to operational or real-time applications.In this study, we have introduced a comparative framework for medical image segmentation with traditional, swarm intelligence and convolutional neural networks as a deep learning mechanism. This framework helps to design a real-time universal medical diagnosis system for various types of images such as MRI for brain data analysis, dermoscopic for skin, microscopic for blood leukemia, and CT-scan images for lungs. Here, we present a comparative study using six different scenarios such as FCM,"} +{"text": "Since the onset of the COVID-19 pandemic, there have been drastic changes to people\u2019s day-to-day lives. Following the 2020 U.S. national shutdown, day-to-day life included increased social isolation, COVID-19 anxieties, disruptions to employment, and limited access to staple goods for many. Compared with the general population, the impacts of these changes are much less understood for the aging population, and particularly for older adults with a disability or chronic disease, who are at greater risk of adverse effects from COVID-19 and related circumstances. Drawing on the breaking results of the AARP Vital Voices national survey (n=2071), conducted throughout the pandemic (2019 to 2021), this poster presentation sheds light on the pandemic experiences of older American adults with and without disability or chronic disease. Interesting findings are discussed across age groups and by gender. Overall, the results show notable differences in the social connection experiences of older adults with a disability or chronic disease (n=571) pre-and-post the national shutdown, along with differences in the accessibility of household items, staple foods, and medical care in the wake of the 2020 U.S. national shutdown. The implications of these findings are discussed in the context of related literature."} +{"text": "The role of the physiotherapist is vital in the recovery of post-COVID-19 patients, but fear of contagion is a possible feeling among healthcare professionals. The objective of this study is to assess the mental health effects that COVID-19 has had on healthcare workers, including rehabilitation care, in times of pandemic.A systematic review was conducted using the PRISMA format in the Pubmed, SCOPUS, and Web of Science databases between July and September 2022. Keywords included were \u201chealthcare providers,\u201d \u201cCOVID-19,\u201d \u201cMental Health,\u201d and \u201cPsychological Distress.\u201d Methodological quality was assessed using the Joanna Briggs Institute critical appraisal tools.A total of 14 studies were included in this review. The study population was healthcare professionals including the rehabilitation services. In total, 4 studies reported exclusively on anxiety and stress levels in physiotherapists providing care during the pandemic.The mental health of healthcare professionals has been compromised during the pandemic. However, initially, research was only focused on physicians and nurses, so the need arises to include those professionals, such as physiotherapists, who are also in direct contact with COVID-19 patients.https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=367664, identifier: CRD42022367664. COVID-19 is a disease caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) .For COVID-19 to be declared a pandemic in March 2020, it had to show alarming levels of spread and severity affecting a large number of people, as well as outbreaks in more than one continent. Some of the consequences of SARS-CoV-2 implied limitations in participation and restrictions in access to different care spaces and services. Many of the diagnosed patients and others with different pathologies began to receive care through online appointments, leaving face-to-face consultations for more serious cases in order to combat the onslaught of the disease and its spread , 5. In aCOVID-19 also has a serious impact on people's mental health . PsycholThe Job Demands-Resources (JD-R) model, in contrast to the theories of job design and job stress, highlights the role of job stressors, being used to predict burnout, engagement, and additionally, to identify the consequences of sickness absenteeism and job performance. With the JD-R model, it is possible to explain, understand, and predict employees' wellbeing. According to the theory, work environments can be divided into job demands and job resources, and this can be applied to all occupations. However, there are job demands and job resources specifically relevant to each occupation or profession . Job demAccording to a systematic review, some of the risk factors most associated with psychological distress during the COVID-19 pandemic were being female, from lower socioeconomic status , belonging to rural areas, and those at higher risk of COVID-19 infection . These population groups showed a higher prevalence of suffering episodes of depression and anxiety compared to other groups . In factThere is the fact that research during this time of pandemic has yielded significant outcomes in different areas, being one of them healthcare professionals. However, it is understood that \u201cthose in the front line\u201d are only physicians and nurses , leavingHealthcare professionals play a major role in the care and contact with people with COVID-19, many of them being part of the first line of defense against the virus . They maDue to the impact of the pandemic on all health services, a restructuring of rehabilitation services was initiated as Physical Therapy areas were transformed into temporary hospitalization rooms. In fact, the Spanish Society of Physical Medicine and Rehabilitation , publishThe aim of this study was to assess the effects that COVID-19 has on the mental health, i.e., the psychological distress, of healthcare workers of the rehabilitation services when caring for patients in times of pandemic.A systematic review was conducted following the guidelines of the PRISMA statement thesaurus was consulted, yielding the descriptors health personnel and physiotherapists, mental health, physiological stress, anxiety, depression, and COVID-19. In order to enlarge the scope of the search, synonymous terms were used to complete the search based on the Medical Subject Headings (MeSH) descriptors , linked Original articles, including meta-analyses, systematic reviews, cohorts, cross-sectional, and case-control studies published in English and Spanish were included in this review.The following criteria were used for the selection of articles:Original articles published in English and Spanish.Articles published from 2020 to date.Type: original articles, meta-analysis, case reports.Articles measuring any of the following values and/or effects: level of depression, level of stress and level of anxiety, number of cases of professionals with depression, stress and/or anxiety, comparison of levels before vs. during the COVID-19 pandemic, and comparison according to country or type of profession/service.Studies that did not meet the previously established inclusion criteria, that did not answer the research question, or that were not related to the objective of the review.Studies of low scientific-technical quality after applying the quality assessment tool.Study population other than healthcare professionals and which did not include rehabilitation professionals.Typology: opinion articles, commentaries, editorials and letters to the editor/head, and quasi-experimental.Initially, two researchers independently carried out the searches, as set out in the search strategy for each of the chosen databases. Subsequently, one researcher eliminated duplicate articles and those that did not meet the previous criteria, and finally included studies accordingly, after reading the titles and abstracts. Subsequently, one author reviewed the full text of the potential studies for the review and made the decision to include or exclude them. Discrepancies were resolved by the first two authors.The methodological quality of the selected studies was determined using the critical appraisal tools of the Joanna Briggs Institute (JBI) of the University of Adelaide . The purThe initial search strategies identified a total of 490 references, which were screened according to the topic of this review. A total of 14 articles were finally selected , 11 of wThere was a variety of countries identified in the studies, among them, two were conducted in Spain, two in Brazil, two in Italy, and one in other countries such as Portugal, Korea, Austria, Indonesia, and Egypt, among others. In relation to the sample, in 6 of the 14 studies included in the review, the sample was exclusively composed by physiotherapists.The included articles were assessed with the JBI critical appraisal tool, where both quantitative and qualitative studies obtained medium-high scores.To assess anxiety levels, the different studies used the following scales: GAD-7, DASS-21, and HADS. The Generalized Anxiety Disorder tool (GAD-7) is a self-administered screening test designed to identify probable cases and severity of anxiety. The GAD-7 is used in adults >18 years old and includes 7 items on a Likert-type scale . Scoring ranges from 0 to 21, with scores of 5, 10, and 15 set as cut-off points for mild, moderate, and severe anxiety, respectively. Further assessment is recommended when the score is 10 or higher .The level of anxiety can also be assessed with the DASS-21 \u201cAnxiety subscale,\u201d which has been one of the preferred assessing instruments during the COVID-19 pandemic , 37, 38.The Center for Epidemiologic Studies Depression Scale (CES-D), the Patient Health Questionnaire depression module (PHQ-9), the Depression Anxiety Stress Scales , the Hamilton Depression Scale (HAMD-17), and the Self-rating Depression Scale (SDS) are avaiThe HADS is one of the scales used in the assessed studies to evaluate depression and anxiety. It had been used before with nursing staff in Poland , 39. It In addition, the level of depression was qualitatively assessed through semi-structured interviews to record and code the emotional experiences of healthcare professionals, including the rehabilitation services, in times of pandemic \u201336.The Perceived Stress Scale (PSS-10) consists of 10 questions about feelings and thoughts during the last month. Responses are given for each question on a 5-point scale which ranges from \u201cnever\u201d to \u201cvery often.\u201d Then, the total is calculated. Scores ranging from 0 to 13 are regarded as low stress; scores from 14 to 26, moderate stress; and between 27 and 40, scores are considered high perceived stress , 25.The 22-item Impact of Events Scale-Revised (IES-R) is used to assess post-traumatic stress symptoms during the past 7 days. Each item is scored from 0 to 4. The total scale score ranges from 0 to 88. Values above the cut-off point of 33 indicate a clinically relevant symptom , 29.All studies included questionnaires covering socio-demographic data . However, some of them included questions related to the health of the participants , 28 and Regarding mental health, some studies , 34, 36 The Maslach Burnout Inventory (MBI), on the other hand, assesses the level of burnout. It is composed of 22 items designed to evaluate the three dimensions of burnout: Emotional Exhaustion (nine items); Depersonalization (five items); and Personal Accomplishment (8 items). All MBI items are scored using seven-level frequency ratings, from \u201cnever\u201d (=0) to \u201cevery day\u201d (=6). Burnout is confirmed by obtaining high scores on the subscales that assess emotional exhaustion (0\u201354 items) and depersonalization (0\u201330 items) and low scores on the Personal Accomplishment subscale (0\u201348 items) , 31, 33.The WHOQOL measures health-related quality of life. It was assessed using the Polish version of the abbreviated World Health Organization instrument (WHOQOL BREF). It has 4 domains: D1-Physical; D2-Psychological; D3-Social Relationships; and D4-Environmental, and consists of 26 questions. The respondents rate each aspect on five-point Likert scales. The domain score reflects an individualized perception of each quality-of-life domain, and it is scaled in a positively framed direction: the higher the score, the higher the health-related quality of life , 31.The COVID-19 pandemic has radically led to a change in lifestyle, affecting different aspects . Newly pBut not everything was bad. I have learned a lot\u201d (Regarding the qualitative assessment, three of the analyzed studies have something in common \u201336, named a lot\u201d . A Spanid a lot\u201d . Anotherd a lot\u201d .One of the qualitative studies , two quaOn the other hand, most of the quantitative studies, with the exception of one , followeFar\u00ec et al. concludeAs for the differences found between types of healthcare professionals, only Szwamel et al. showed tThis study allowed to examine professionals in the area of rehabilitation as an important part of health care during the pandemic. Including this population in the investigation and carrying out research to generate new interventions in mental health are the strongest parts of this research. Likewise, the assessment of rigor and methodological quality of the included studies, and their variables, permit to support solid conclusions and generalizations. Despite the results of interest provided in this research, it would be pertinent to continue deepening the subject of study.The present study shows some limitations. Firstly, it should be noted that one article written in German was rejected, as no translation could be found, so it is possible that some articles that met the rest of the inclusion criteria were left out for this language reasons. In addition, eleven articles were rejected for not having the exact study population, i.e., only included physicians and nurses but not rehabilitation professionals, or there was not a clear statement about their inclusion in the study. The vast majority of studies was also found to not show strategies to control for confounding factors, except for two articles that do mention this aspect.Some of the studies did not show a balance between men and women, so it was not possible to assess sex differences related to the variables described in the objective. On the other hand, certain articles did not include a variety of professional groups that would allow establishing differences between professionals/services. Therefore, the findings may have a limited possibility for generalization to all healthcare professionals as the studies only considered physiotherapists to study the professionals of rehabilitation services, and did not include other groups of important professionals, i.e., occupational therapists, speech therapists, etc. Likewise, although there is a variety of countries in the total number of studies, the quantity is not sufficient, and therefore, the representativeness of the results found cannot be extrapolated to the rest of the health professionals who carry out their healthcare work in the rest of the countries of the world.Professionals of the rehabilitation services indicated that the quality of services has been affected by COVID-19, compromising the effectiveness of care , 42. ForThe use of technology is a good strategy for communication and medical intervention, including rehabilitation. In this sense, it is also important to carry out studies that compare results between professionals in the same health care area, i.e., not only taking into account physiotherapists as the only ones involved in rehabilitation, though this may not be applicable in all cases, as telerehabilitation allows contact to be maintained without fear of contagion. Technology applied to medicine may also empower the patient in their treatment to become an active participant in their recovery, and would also enable the caregiver to assume their role while avoiding overload and being supportive in the process of rehabilitation of the patient, without requiring the continuous presence of the professional in charge.Equally, the information on the psychological impact of the pandemic throughout the last 2 years contributes to expand knowledge and increases the interest on intervention strategies focused on the health worker's mental health, including professionals in the rehabilitation area. These interventions can be designed to modulate or reduce the risks and consequences of mental health deterioration, as part of a method of prevention of occupational diseases.Mental health of healthcare professionals, in general, has been compromised as the COVID-19 pandemic has progressed, compared to before the onset of the pandemic. Women were also found to be more likely to suffer increased levels of anxiety, burnout, and depression, and professionals with children and families showed higher levels of distress and anxiety in caring for patients with COVID-19. Additionally, professionals who were in the front line of the battle against the virus have seen their mental health compromised but with values below those of the general population.Changes in working hours and care settings, patient overload, fear of becoming infected and infecting loved ones and/or patients, among others, may be precipitating factors for an alteration in the mental health of healthcare professionals in times of the COVID-19 pandemic. Such an alteration can be a major problem at a personal, family, and professional level and can increase the risk of professional malpractice.The original contributions presented in the study are included in the article/Conceptualization: SB-B, RA-C, JG-S, CM-L, JG-I, JF-R, and CR-F. Data curation: SB-B, JG-I, and RA-C. Formal analysis: SB-B, RA-C, JG-S, CM-L, CR-F, JG-I, and CR-F. Investigation: SB-B, RA-C, JG-S, CM-L, JG-I, and JF-R. Methodology: SB-B, JG-S, CM-L, CR-F, and JF-R. Project administration: JG-S and CR-F. Resources: RA-C, JG-S, CM-L, JG-I, and JF-R. Software: SB-B, RA-C, JG-S, and JF-R. Supervision: JG-S, JF-R, JG-I, and CR-F. Validation: RA-C, JG-S, CM-L, and CR-F. Visualization: JF-R and JG-I. Writing\u2014original draft: SB-B, RA-C, JG-I, and JG-S. Writing\u2014review and editing: JG-S, CM-L, JF-R, and CR-F. All authors contributed to the article and approved the submitted version."} +{"text": "In the published article, there was an error in the Funding statement, which was mistakenly included as an Acknowledgements. The corrected statement appears below:This project was funded by the Deanship of Scientific Research (DSR), King Abdulaziz University, Jeddah, under grant no. (D:830-1021-1443). The authors, therefore, gratefully acknowledge DSR\u2019s technical and financial support.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way."} +{"text": "Trypanosoma cruzi transmission, due to the zoonotic nature of most transmission cycles, with well over 100 vector species widely spread across the Americas. Then, Rojas de Arias et al.T. cruzi transmission and (2) provide an overview of some innovative approaches that, I believe, can play important roles in the development and operation of stronger control-surveillance systems for vector-borne CD.The multinational initiatives for the control/surveillance of Chagas launched by disease-endemic countries and the Pan American Health Organization-World Health Organization (PAHO-WHO) contributed to control house-infesting triatomine-bug populations and to reduce disease incidence. However, after 30 years, Chagas disease (CD) transmission persists the Americas. In their recent review, Rojas de Arias et al.eradication - the complete elimination of an infection, with no new cases recorded in the absence of control measures. Eradication is practically impossible for zoonoses such as CD, whose etiological agent can be transmitted by 150+ vector species and infects a wide range of wild vertebrate hosts from the USA to Patagonia.Disease control needs clear goals, and those goals often heavily depend on the natural history, risk factors, transmission routes and dynamics, pathogenesis, and treatment of the disease. Eradication, elimination, reduction of incidence, reduction of the number of severe cases, and reduction of fatality rates are all possible goals of disease control programs.elimination of the strongly synanthropic, non-native populations of a few \u201cprimary\u201d vectors - mainly Triatoma infestans and Rhodnius prolixus.R. prolixus from Mexico and some countries of Central America was certified in 2011 by PAHO/WHO, but R. prolixus-infested houses were detected in rural sites in Mexico after certification.T. infestans in Mexico was recorded after 50 years.T. infestans populations were not controlled in Arequipa regularly offers online courses on infectious-disease prevention. Online courses can reach many more people than traditional courses; for example, more than 20,000 health agents have already taken the Surveillance and Control of Vectors of Importance in Public Health course (https://www.unasus.gov.br/cursos/curso/45783), which covers CD vectors and was recently revised and expanded in a Spanish version. There is also the prospect of producing an English version to reach the USA and possibly other English-speaking countries with native triatomines . Recently, another UNASUS course was opened focused on CD in primary health care (https://www.unasus.gov.br/cursos/curso/46776). The Brazilian Ministry of Health also offers courses in parasitology, chemical vector control, and triatomine-bug identification once or twice a year . Globally, a web-based search identified over 140 courses on vector biology and vector-borne diseases.II - Development of apps to identify vectors and improve surveillance with community participation and citizen science,An important topic in the training of heath agents working with CD is the identification of triatomines. This is because surveillance and control actions depend on which species occur in the area; the correct identification of triatomines is therefore critical for CD control-surveillance. Triatomines are traditionally identified using printed dichotomous keys based on morphological characters, but new electronic keys are available and can run on smartphones.Bug notification by householders enhances vector detection and can be more effective than either active searches by control agents or the deployment of vector-detection devices.III - Vector-borne transmission-risk mappingT. cruzi transmission was assessed based on socioeconomic, demographic, entomological, and environmental indicators.Flexible entomological-risk indicators that cover native and non-native vectors are needed to support local decision-making. The identification of areas with greater vulnerability to the occurrence of vector-borne CD is essential to prevention, control, and surveillance activities. Using ecological-risk maps based on vector distribution models and disease-risk maps based on incidence data, Sarkar et al.T. cruzi transmission\u2019 and emphasise that stronger surveillance systems are needed to monitor and control CD. I recognise that much has been done to control CD in the Americas,,,,In summary, I fully agree with Rojas de Arias et al.Trypanosoma cruzi transmission. Mem Inst Oswaldo Cruz. 2022; 117: e210130.Comments on the article: de Arias AR, Monroy C, Guhl F, Sosa-Estani S, Santos WS, Abad-Franch F. Chagas disease control-surveillance in the Americas: the multinational initiatives and the practical impossibility of interrupting vector-borne"} +{"text": "Mitochondrial diseases are clinically heterogeneous, can occur at any age, and can manifest with a wide range of clinical symptoms. They can involve any organ or tissue, characteristically involve multiple systems, typically affecting organs that are highly dependent on aerobic metabolism, and making a definitive molecular diagnosis of a mitochondrial disorder is challenging.Clinical data of the proband and his family members were gathered in a retrospective study. Whole-exome sequencing and full-length sequencing of the mitochondrial genome that were performed on peripheral blood, urine, and oral mucosa cells were applied for genetic analysis.In this study, we reported a childhood-onset mitochondrial phenotype in a 13-year-old patient. Analysis of the next-generation sequencing data of the nuclear genome and the full-length sequencing of the mitochondrial genome revealed the rare m.10000G>A variant in MT-TG that was present at variable heteroplasmy levels across tissue types: 32.7% in the blood, 56.15% in urinary epithelial cells, and 27.3% in oral mucosa cells. No variant was found in the peripheral blood of his mother and sister. No pathogenic mutation of nDNA was found.de novo m.10000G>A variation in the highly conserved sequence of MT-TG appears to suggest a childhood-onset mitochondrial phenotype in the 13-year-old patient, thus broadening the genotypic interpretation of mitochondrial DNA-related diseases.Our results added evidence that the Mitochondrial diseases are a group of genetic disorders that are characterized by defects in oxidative phosphorylation and are caused by mutations in genes in the nuclear DNA (nDNA) and mitochondrial DNA (mtDNA) that encode structural mitochondrial proteins or proteins involved in mitochondrial function . Mt-tRNAOne patient was included in the study. The patient was managed at the Department of Neurology, Hunan Children's Hospital. The parents of the patients provided written informed consent. This study was approved by the Medical Ethics Committee of Hunan Children's Hospital.The whole-exome sequencing method was performed according to our previous research methods . SequencFull-length sequencing of the mitochondrial genome of peripheral blood, urine, and oral mucosa cells was performed according to our previous research methods . Full-leNC_012920.1) in MT-TG . White blood cell count, glucose and protein content, microbiological tests, and immune encephalitis-related antibody tests on the cerebrospinal fluid were normal. Color Doppler echocardiography examination showed that the aorta and the ascending aorta were widened, and that left ventricular systolic function was normal. Nerve conduction studies and electromyography suggested axonal and demyelination involvement of peripheral neuropathy. Brain MRI showed bin MT-TG that wasin MT-TG . The impThe clinical features of this patient include hypothyroidism, exercise intolerance, weakness of the limbs, hearing loss, encephalopathy, neuropathy, hyperlactemia, and typical MRI changes. According the mitochondrial diagnostic criteria (MDC) , the tothttp://www.mitomap.org). Among them, only 1 variant, m.10010T>C, is considered \u201cdefinitely pathogenic.\u201d The known pathogenic variant 10010T>C on the same tRNA was detected in multiple patients, one of which was 15% in skin fibroblasts, 17% in blood and >90% in skeletal muscle . and his healthy parents and older sister did not carry the de novo variant. Second, the patient's phenotype was consistent with mitochondrial disease, and no pathogenic mutation of nDNA was found. Third is the variable heteroplasmy levels across tissue types. Fourth is that hearing loss, encephalopathy, and neuropathy are common clinical features of the reported MT-TG mutant disease , and minor(s)' legal guardian/next of kin, for the publication of any potentially identifiable images or data included in this article.HY: conducted the literature review and drafted the manuscript. VZ, LA, and SG: make substantial contributions to conception and interpretation of data. LW: revised the manuscript critically and have given final approval of the version to be published. All authors contributed to the article and approved the submitted version.This study was supported by a grant from the National Natural Science Foundation of China (Grant No: 81671297).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Although sleep hygiene is a well-studied factor of good sleep , less is known about its role in the complaints on anxiety and depression .The aim was to reveal direct and indirect effects of sleep behavior on subjective sleep quality, anxiety and depression.174 people aged 17-57 without diagnosed sleep disorders filled the Scale of Behavioral Factors of Sleep Disturbances , Insomnia Severity Index , Hospital Scale of Anxiety and Depression , Beck\u2019s Anxiety and Depression Inventories .Taking medications and non-medications before sleep, alcohol, tonic drinks and using gadgets in the evening, delaying bedtime, self-limitations after poor nights, poor adherence to the regimen and postponement of the morning rise were characterized by an indirect effect on anxiety and depressiveness through poor sleep . Self-limiting behavior and delaying the morning rise are associated with higher levels of anxiety and depression, even in the absence of sleep-related complaints .Based on the data we suggest that the dysfunctional role of behavior on anxiety and depression is predominantly indirect (through the perpetuation of complaints), but it can also be direct (regardless of complaints of sleep disorders). Research is supported by the Russian Foundation for Basic Research, project No. 20-013-00740.Research is supported by the Russian Foundation for Basic Research, project No. 20-013-00740."} +{"text": "The National Institute of health (ISS), within the G20 Health Working Group, launched at March 2021 an innovative training Laboratorium, to strengthen the capacity and competencies of the Public Health Workforce (PHW) on prevention, preparedness and response to health crises. It was recognised in the G20 Declarations of Ministers of Health and Leaders and it was aimed to the development of training tools suitable for distance learning. ISS proposed the design of e-Learning courses based on an integration of WHO\u2019s Competency-Based Education (CBE) and Problem Based Learning (PBL) models.Describe the design of a pilot e-Learning course based on CBE- PBL models, focused on Epidemic Intelligence (EI).https://www.eduiss.it. By the end of the course participants will be able to evaluate the potential use/applicability of EI systems, focusing on event-based surveillance for preparedness and early warning at country/institutional level. CBE is the basis for articulating the outcomes and identifying the required competencies and PBL for creating learning activities associated to learning outcomes. Interactive tools are associated to learning objectives. The course is offered to public health professionals free of charge, in English language. About 16 hours are required to complete all the activities and receive attendance certification.The pilot e-Learning course \u201cUse of Epidemic Intelligence systems with a particular focus on event-based surveillance for pandemic preparedness\u201d will be delivered by ISS e-Learning platform Continuously updated training of PHW is a hallmark to better face health challenges. In our proposal, the learning methodology integrates CBE vision with PBL approach, adapting both to e-Learning context.Pietro Carbone, Debora Guerrera, Federica Regini, Licia Bacciocchi, Stefania Bocci, Silvia Stacchini, Giovanna Failla - ISS, Rome Italy\u2022\u2002Design an innovative e-learning course through synergic integration of two educational models crucial in public health, CBE and PBL.\u2022\u2002Adapt e valorize CBE and PBL models in the context of e-Learning."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.5Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.13The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.14Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.18The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Author list. Author \u201cZe Jin\u201d was erroneously excluded. The corrected Author list appears below:In the published article, there was an error in the \u201cInseok Hwang, Ze Jin, Je-Wook Park, Oh-Seok Kwon, Byounghyun Lim, Jisu Lee, Hee-Tae Yu, Tae-Hoon Kim, Boyoung Joung, Hui-Nam Pak.\u201dIn the published article, there was an error in Author Contributions were described incorrectly:In the published article, the \u201cIH contributed to the data, statistical analyses, and writing of the manuscript. J-WP contributed to the statistical analyses and data acquisition. O-SK contributed to the software programming and data acquisition. BL confirmed the data acquisition and references. JL provided the support for the software programming. ZJ contributed the clinical data acquisition. H-TY, T-HK, and BJ contributed to the clinical data acquisition and interpretation of clinical data. H-NP contributed to the study design, clinical data acquisition, data interpretation, and revision of manuscript. All authors contributed to the article and approved the submitted version.\u201dAuthor Contributions statement appears below:The corrected \u201cIH and ZJ contributed to the data, statistical analyses, and writing of the manuscript. J-WP contributed to the statistical analyses and data acquisition. O-SK contributed to the software programming and data acquisition. BL confirmed the data acquisition and references. JL provided support for the software programming. H-TY, T-HK, and BJ contributed to the clinical data acquisition and interpretation of clinical data. H-NP contributed to the study design, clinical data acquisition, data interpretation, and revision of manuscript. All authors contributed to the article and approved the submitted version.\u201dThe authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The morbidity and mortality of cardiovascular diseases (CVD) in patients with rheumatoid arthritis (RA) is significantly higher than those in the general population, leading to RA-related CVD has attracted broad attention and numerous articles have been published. However, no study has systematically examined this area from a scientometric perspective. This study aimed to visualize the knowledge structure and identify emerging research trends and potential hotspots in this field.Articles and reviews on RA-CVD published from 2001 to 2021 were extracted from the Web of Science Core Collection database. CiteSpace and VOSviewer software were used to visualize the knowledge network of countries, institutions, authors, references and keywords in this field. SPSS and Microsoft Excel software were used for curve fitting and correlation analysis.A total of 2,618 articles and reviews were included. The number of publications about RA-related CVD significantly increased yearly. Publications were mainly concentrated in North America, Europe and East Asia. The United States contributed most with 699 publications, followed by the United Kingdom and Italy. Gross Domestic Product was an important factor affecting scientific output. University of Manchester and Professor Kitas George D. were the most prolific institutions and influential authors, respectively. Journal of Rheumatology was the most productive journal for RA-related CVD research. The research hotspots switched in the order of clinical features (cardiovascular events), mechanism exploration, anti-tumor necrosis factor therapy, risk factors, and antirheumatic drug safety, which can be observed from the keyword analysis and co-cited reference cluster analysis.This study found that research on RA-related CVD is flourishing. The safety and cardiovascular pharmacological mechanisms of anti-rheumatoid drugs, especially targeted synthetic DMARDs, would be the focus of current research and developmental trends in future research. Rheumatoid arthritis (RA) is a systemic autoimmune disease with an approximately 0.5\u20131% prevalence worldwide . As repoGiven the above aspects, RA-related CVD has received increasing attention from scholars. However, there are still some controversies surrounding RA- related CVD, such as CVD risk factors, prevention drugs, and optimal treatment protocols . MotivatNew researchers can benefit from an overview of the knowledge structure and current hotspots in a given field \u201313. ScieWeb of Science (WoS) contains more than 12,000 international academic journals, which is one of the most comprehensive and authoritative database platforms for accessing global academic information and is widely used for scientometric analysis , 12. A tData extraction was performed by two independent researchers to ensure the accuracy and reliability of the results. Any disagreements between the two researchers were discussed until a consensus was reached. The extracted data included authors, titles, publication years, citation times, countries, institutions, journals, highly cited articles, references, and keywords. The Hirsch index (H-index) was obtained by WoS. In addition, taking into account the differences in economic and demographic conditions in different countries, several ratio indexes were introduced, including the number of papers per million people and the number of papers per trillion Gross Domestic Product (GDP) .P < 0.05 was defined as statistically significant.Statistical analysis was performed using IMB SPSS 22.0 , and Microsoft Excel 2016 . Statistical plots were drawn using OriginPro 9.1 . Categorical data were expressed as counts (percentages). The correlation strength between continuous variables was assessed by Pearson\u2019s correlation coefficient. Visualization software including VOSviewer, CiteSpace and Scimago Graphica were used in the scientometrics analysis. Scimago Graphica software was used to map the global distribution of national publications and the cooperation network of countries. VOSviewer software was used to visualize the co-authorship analyzes of institutions and authors, as well as the co-occurrence analysis of keywords. CiteSpace was mainly used for reference co-citation analysis, keyword burst detection, and drawing timeline views of reference clusters.2 = 0.9849).After the literature screening, a total of 2,618 publications were included in the final analysis, including 2201 original articles and 467 reviews. The detailed distribution of annual publications for RA-related CVD research was shown in r = 0.221, p = 0.077), while there was a high positive correlation between the number of publications and GDP .From the world map of country contributions , it can As shown in 1 in the United States. The Karolinska Institutet in Sweden had the highest average citation of 102.3. Mayo Clinic had the highest H-index of 46.It was roughly estimated that more than 2,000 institutions contributed to this field. During the last two decades, a total of 514 academic journals published papers on RA-related CVD. In terms of the top 10 prolific authors , Kitas GA visualization of the author\u2019s co-authorship analysis was generated by VOSviewer software . It can The references in the co-cited network were divIn the keyword analysis, the meaningless keywords were excluded and the same meaning keywords were merged. This study provided a scientometric analysis of RA-related CVD research from 2001 to 2021 to find milestone achievements and predict new research hotspots. It can also help beginners intuitively and systematically understand the development process and trends in this field. This study found that the number of publications and citations of RA-related CVD have been numerous and increasing in the past two decades . AccordiFrom the perspective of national contribution, the United States ranked first in the world in terms of publications, citations and H-index, indicating its dominant influence . Four ofPublishing research results in international peer-reviewed journals is an important part of establishing effective scientific communication . The anavia accentuation of established and novel risk factor pathways. By implication, long-term suppression of the systemic inflammatory response in RA should effectively reduce the risk of CVD could effectively inhibit inflammation and reduce the risk of RA-related CVD \u201340. BroaTimeline plots of clusters can track the evolution of research hotspots and predict the research trends in the coming years . In addiTaken together, this study has described the research status, hotspots and further trends in the RA-related CVD. Researchers, especially newcomers, could benefit from this study as they could clearly understand the basic knowledge structure of the field, including countries, institutions, authors and journals, and be inspired by the research hotspots and frontiers. We have to acknowledge that this study has some inherent limitations in scientometrics. First, the databases are constantly updated and only the WoSCC database was analyzed in this study. Some articles published in other databases may be omitted. However, as described in previous studies, WoSCC was the most commonly used and sufficiently large database for scientometric analysis to reflect the current status in a given field , 16, 17.To our knowledge, this was the first study to conduct a comprehensive scientometric analysis of global publications on RA-related CVD from 2001 to 2021. Our findings suggested that RA-related CVD has attracted considerable and growing attention from scholars. Up to now, North American and European countries have been leaders in this field, which is inseparable from adequate funding sources. University of Manchester and Professor Kitas George D. were the most prolific institutions and influential authors, respectively. Journal of Rheumatology and Annals of the Rheumatic Diseases were the most prolific and influential journals in RA-related CVD research, with the largest number of publications and citations, respectively. Cardiovascular events, mechanism exploration, risk factors and antirheumatic drug safety were the hotspots in this field. Of note, the research foci have shifted with time, and the safety and cardiovascular pharmacological mechanism of anti-rheumatoid drugs, especially targeted synthetic DMARDs, are considered to be important research directions in the future. Long-term, robust and well-designed clinical trials deserve further attention.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding authors.PW and PL: writing-original draft. PW and YZ: conceptualization, project administration, and writing-review and editing. PL, BZ, YW, and JC: data curation and methodology. LH, JW, and JC: formal analysis and validation. PW, RL, and JG: visualization and software. All authors contributed to the article and approved the submitted version."} +{"text": "Immigrants in Japan face multiple health care challenges. There is limited research addressing how all-cause mortality differs between foreign residents and Japanese citizens, including the impact of the COVID-19 pandemic. We assessed whether all-cause mortality rates between Japanese citizens and foreign residents living in Japan differ, and whether these differentials changed after the start of the COVID-19 pandemic. We conducted a cross-sectional analysis using vital statistical data of all deaths among citizens and foreign residents that occurred within Japanese borders aggregated every 6 months between January 1, 2015 and June 30, 2021. Data were used to calculate sex-, region-, and 20-year age group-specific standardized mortality rates using the direct method based on the population distribution of Japanese citizens in 2021 by sex, region, and 20-year age groups. Chi-squared tests and linear regression were used to assess whether the pandemic was associated with changes in mortality rates among groups and changes in the mortality differentials between citizens and non-citizens, respectively. All-cause mortality increased monotonically with age for men and women. Men had higher mortality than women, regardless of age or nationality. All-cause mortality is lower among immigrants than Japanese citizens between the ages of 20\u201359, but higher under the age of 20 and over the age of 59. The pandemic was associated with significant changes in mortality in most groups, but no statistically significant changes in the mortality differentials between immigrants and Japanese citizens were detected. Young immigrants are generally healthier than their Japanese counterparts, in line with the healthy migrant hypothesis. Younger migrants are at higher risk of mortality, possibly due to increased vulnerability to psychologic stress. Older migrant mortality converged with citizen mortality, consistent with acculturation that occurs with longer duration of residence. The pandemic did not exacerbate health inequities for foreign residents with respect to mortality. \u2022Foreign residents have lower mortality than citizens between the ages of 20\u201359.\u2022Foreign residents have higher mortality than citizens <20 and >59 years of age.\u2022COVID-19 did not change mortality differentials between immigrants and citizens. Most imIn particular, there is significant concern that the COVID-19 pandemic will exacerbate already existing health inequities between foreign residents and citizens . ImmigraJapan's economy was also considerably impacted by the pandemic. Immigrant workers are particularly vulnerable to economic shocks, and the pandemic affected many of the sectors in which foreigners are concentrated: construction, food-production, hospitality, and healthcare, among others . COVID-1Historically, immigrants residing in Japan have faced challenges within the health care system, including language barriers and difficulty enrolling in universal health coverage . LanguagFurthermore, some clinics may be reluctant to accept non-Japanese patients, particularly those without insurance; when they are accepted, they may be charged higher fees or discharged earlier than dictated by standard care . Fear ofTo the best of our knowledge, only one study \u2013 written in Japanese using 2010 summary data \u2013 has assessed immigrant all-cause mortality in Japan to date, finding that foreign resident mortality is less than that of their Japanese counterparts in younger age groups but switches to being greater among the elderly . FurtherGiven that the immigrant population is only expected to rise in Japan , further22.1zairy\u016bk\u0101do), such as for tourism, and only includes deaths that occurred within Japan's borders.We used daily and individual-level mortality data obtained from the vital statistics of the Ministry of Health, Labour and Welfare of Japan (MHLW) between January 1, 2015 and June 30, 2021. This data does not include those who stay in Japan for a short period of time without a residency card , sex, age group , and region . Thus, the mortality rates in this study are not annual rates, as they are estimated for each half year. The number of Japanese citizens and foreign residents in any given year was obtained from population data in the Basic Resident Register . The sta2.3p (\u03c72)\u2019 in The COVID-19 pandemic was defined as beginning in January 2020 when the first official case was identified in Japan. The standardized mortality for any given sex, age, and citizenship group was compared before and after the COVID-19 pandemic using the chi-squared test; testing was performed by constructing a two-by-two table with pre-COVID-19 and post-COVID-19 mortality. The results of these tests are reported as \u20182.4To check whether the difference in mortality between Japanese citizens and foreign residents within a given sex- and age-stratified group changed significantly during the COVID-19 pandemic, we conducted linear regression with the outcome variable defined as the difference between citizen and foreigner mortality rates. The following covariates were included in the model: time, a COVID-19 dummy variable (pre-vs post-January 2020), and their interaction term. Time was encoded as a variable such that the first half of 2015 was set to 2015.0, the second half was set to 2015.5, and so on until the first half of 2021 (2021.0). The dummy variable was defined as 0 pre-COVID-19 and defined as 1 for time points after the start of the pandemic . The statistical significance of the interaction term, which was tested with t-tests, was used to determine whether there was a change in the mortality differential between the two groups before and after the start of the pandemic. The results of these tests are shown in 33.1Standardized mortality rates stratified by sex and age group are shown in 3.2The results of the chi-squared analyses for changes in mortality trends are shown in 4Using national Japanese mortality data, we assessed trends in mortality between 2015 and 2021 in Japanese citizens and foreign residents. We found that among those aged 19 or less and 60 and over, immigrants had higher mortality than their Japanese counterparts, but that these trends reversed among those aged 20\u201359 years. Furthermore, our results suggest that though mortality rates changed after the start of the pandemic, the mortality differentials between foreign residents and Japanese citizens did not change significantly.healthy migrant effect, in which those who migrate represent a healthier subpopulation of their native country , come with their parents, or participate in internships and educational opportunities. Children, in particular, may suffer if their parents are not able to take full advantage of local health care/insurance systems, do not have stable employment, or are limited in providing safe housing and food security , and the 10.13039/501100001700Ministry of Education, Culture, Sports, Science and Technology of Japan (21H03203). The funding sources had no role in the study design, data collection, data analysis, data interpretation or preparation of the manuscript.The present work was supported in part by a grant from the Ethical approval was granted by the ethics committee of the National Institute of Infectious Diseases, under authorization number 1174.Cyrus Ghaznavi: Conceptualization; Methodology; Software; Validation; Formal analysis; Investigation; Writing - Original Draft; Writing - Review & Editing; Visualization. Akifumi Eguchi: Conceptualization; Methodology; Software; Validation; Formal analysis; Investigation; Writing - Original Draft; Writing - Review & Editing. Yuta Tanoue: Conceptualization; Methodology; Software; Validation; Formal analysis; Investigation; Writing - Original Draft; Writing - Review & Editing. Daisuke Yoneoka: Conceptualization; Methodology; Software; Validation; Formal analysis; Investigation; Writing - Original Draft; Writing - Review & Editing. Takayuki Kawashima: Conceptualization; Methodology; Software; Validation; Formal analysis; Investigation; Writing - Original Draft; Writing - Review & Editing. Motoi Suzuki: Resources; Data Curation; Writing - Original Draft; Writing - Review & Editing; Visualization; Supervision; Project administration; Funding acquisition. Masahiro Hashizume: Resources; Data Curation; Writing - Original Draft; Writing - Review & Editing; Visualization; Supervision; Project administration; Funding acquisition. Shuhei Nomura: Conceptualization; Methodology; Software; Validation; Formal analysis; Investigation; Resources; Data Curation; Writing - Original Draft; Writing - Review & Editing; Visualization; Supervision; Project administration; Funding acquisition.The authors declare that they have no competing interests or financial disclosures."} +{"text": "Due to miscommunication, in the original publication there is a discrepancy in the project number and funding institution, located in Funding Information and Acknowledgement . The corOriginal:Funding: Deputyship for Research & Innovation, Ministry of Education in Saudi Arabia [Grant number IFPRC-146-141-2020].This should be replaced with:Funding: This research work was funded by Institutional Fund Projects under grant No. (IFPDP-39-22).The authors state that the scientific conclusions are unaffected. All co-authors agree with the content of this correction and wish to apologize for any inconvenience caused for the readers resulting from this error."} +{"text": "To explore the value of PET/MRI, including diffusion kurtosis imaging (DKI), diffusion weighted imaging (DWI) and positron emission tomography (PET), for distinguishing between benign and malignant solitary pulmonary lesions (SPLs) and predicting the histopathological grading of malignant SPLs.max), metabolic total volume (MTV) and total lesion glycolysis (TLG) were calculated. Student\u2019s t test or the Mann\u2013Whitney U test was used to analyze the differences in parameters between groups. Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic efficacy. Logistic regression analysis was used to evaluate independent predictors.Chest PET, DKI and DWI scans of 73 patients with SPL were performed by PET/MRI. The apparent diffusion coefficient (ADC), mean diffusivity (MD), mean kurtosis (MK), maximum standard uptake value compared to the benign group . The MD and ADC were significantly lower, and the MTV and TLG were significantly higher in the high-grade malignant SPLs group than in the non-high-grade malignant SPLs group . In the identification of benign and malignant SPLs, the SUVmax and MK were independent predictors, the AUCs of the combination of SUVmax and MK, SUVmax, MK, MD, and ADC were 0.875, 0.787, 0.848, 0.769, and 0.822, respectively. In the identification of high-grade and non-high-grade malignant SPLs, the AUCs of MD, ADC, MTV, and TLG were 0.729, 0.680, 0.693, and 0.711, respectively.The MK and SUVDWI, DKI, and PET in PET/MRI are all effective methods to distinguish benign from malignant SPLs, and are also helpful in evaluating the pathological grading of malignant SPLs. Lung cancer is one of the most common types of cancer in humans and one of the most common causes of cancer-related death . Lung camax) refers to the highest metabolic value in the focus; it can reflect the lesion metabolic activity. Previously SUVmax and related metabolic parameters showed good performance in identifying benign and malignant lesions of the palatine tonsil and benign and malignant lesions of the lung is a common noninvasive method for evaluating SPLs that provides both metabolic and morphological information . Maximumthe lung \u201310. This effect) , and canG uptake . In addiG uptake , 14. In g cancer , 16. Thimax in PET/MRI is a predictor of overall survival in patients with non-small cell lung cancer can reflect the microstructure of tissues by obtaining information about the diffusion of water molecules. Apparent diffusion coefficient (ADC) has good performance in the detection and characterization of pulmonary nodules or masses , 25. It To the best of our knowledge, the diagnosis of SPLs by DKI combined with FDG-PET has not yet been studied. The main purpose of this study was to explore the value of PET/MRI, including DKI, DWI and PET, in distinguishing between benign and malignant SPLs and the histopathological grading of malignant SPLs.From July 2020 to June 2021, a total of 121 patients with pulmonary lumps or nodules diagnosed by chest CT underwent chest PET/MRI. The inclusion criteria were as follows: 1. The lesions were not treated with invasive treatment, radiotherapy or chemotherapy before PET/MRI examination and 2. No FDG-PET/MRI contraindications were present. The following patients were excluded: 1. No histopathological results were obtained (n=13), 2. The cross-sectional diameter of the solid component of the lesion <10 mm (n = 11), 3. Multiple lesions of the lung (n=15) were present and, 4. Poor image quality or incomplete image sequences were present (n = 9). Ultimately, a total of 73 patients were included in this study was used to reconstruct the image. PET scanning (27 min) was performed in the following sequence: MR-based attenuation correction (MRAC), axial T2WI, axial T1WI, DWI, and DKI and 12-channel phased-array body coil. The tracer was max, MTV and TLG values were calculated. SUVmax is the highest SUV value of the lesion as a whole, and MTV is the volume calculated by adding all the voxels larger than the threshold value by setting the threshold of 40% SUVmax. TLG is determined by the multiplication of MTV with average SUV. The parameters of DWI and DKI were measured by two radiologists , blinded to the clinical and pathological information as well as each other\u2019s outcomes, who analyzed all image data separately. When delineating the ROIs, the radiologists were asked to avoid areas of blood vessels, the trachea, necrosis and bleeding to ensure selection of the solid area with uniform texture.All PET, DKI and DWI images were imported into a United Imaging Healthcare (UIH) workstation (uWS-MR: R005) for postprocessing and measurement. The measured values of PET parameters were automatically calculated by the software, which automatically covers the whole lesion, and then the SUV0 is the signal strength of b=0 s/mm\u00b2. The ADC was calculated using two b values and a single exponential fitting model.The parameter values were calculated using the following formula, where b represents the diffusion sensitivity, S (b) is the signal strength at different b values, and SDWI single index ADC calculation formula:where ADC is the apparent diffusion coefficient .2):For the DKI model, the mean diffusivity (MD), mean kurtosis (MK) values are calculated by fitting the following nonlinear equations using four b values on the maximum cross-section of the lesions on TAccording to the histological characteristics of the tumor, lung cancer was divided into three grades: Grade 1 (highly differentiated), Grade 2 (moderately differentiated) and Grade 3 (poorly differentiated). In this study, small cell lung cancer (SCLC) is a highly malignant neuroendocrine tumor; therefore, it is classified as Grade 3; bronchial adenoma is a low-grade malignant tumor, which is classified as Grade 1 in this study. Grade 3 tumors were classified as the high-grade group because of their high malignant degree, while Grade 1 and Grade 2 were classified as the non-high-grade group .All data were statistically analyzed using MedCalc 20.0 software and SPSS 19.0 software . The intragroup correlation coefficient (ICC) was used to evaluate the interobserver reliability of the two radiologists . Indepenmax, MTV and TLG were calculated automatically by workstation software, and there was no need for a consistency check.The parameters obtained by the two surveyors were in excellent agreement. The ICC values of ADC, MD and MK were 0.836, 0.897 and 0.867, respectively. In all subsequent analyses, the average values of the parameters of the two readers were used. The values of SUVmax values of malignant SPLs were significantly higher than those of benign SPLs, while the values of MD and ADC were significantly lower than those of benign SPLs. There was no significant difference in MTV or TLG between the two groups . The values of ADC and MD in high-grade malignant SPLs were significantly lower than those in non-high-grade SPLs, while the values of MTV and TLG were significantly higher than those in non-high-grade SPLs. There were no significant differences in MK and SUVmax values between the high-grade group and the non-high-grade group > AUC (MK) > AUC (ADC) > AUC (SUVmax) > AUC (MD). However, only the difference between AUC (MK+SUVmax) and AUC (MD) was statistically significant (P = 0.0444). In terms of distinguishing between the high-level and low-level groups of malignant SPLs, the AUCs of MD, ADC, MTV and TLG were 0.729, 0.680, 0.693 and 0.711, respectively, but there were no significant differences , and the inclusion standard of lesions was SPLs with the cross-sectional diameter of the solid component of the lesion > 10 mm, which probably reduced the interference of other factors to some extent. In the present study, the high-grade ADC and MD values of malignant SPLs were significantly lower than those of non-high-grade SPLs, which was consistent with previous studies does not significantly improve the diagnostic efficiency of differential SPLs. There was no significant difference in AUCs among single parameters for distinguishing high-grade and non-high-grade lung cancer, which indicated that DWI, DKI and PET were all helpful for the pathological grading of malignant SPLs, but none of those parameters showed higher diagnostic efficiency.In terms of the diagnostic efficiency of distinguishing benign from malignant SPLs, the AUC of SUVThis study has some limitations. First, there were relatively few cases of benign SPLs, squamous cell carcinoma and small cell carcinoma in this study, and there was also a lack of other malignant histological types (such as single metastasis and lung carcinoid). This is due to the nature of prospective studies, where subjects undergoing PET/MRI are usually suspected of having lung malignancy and have a higher incidence of lung adenocarcinoma than other lung cancer subtypes. Second, both DWI and DKI are echo-planar imaging (EPI)-based sequences, which make it difficult to evaluate minimal lesions due to the low signal-to-noise ratio, low spatial resolution, and susceptibility to artifacts of EPI. Third, the ROIs of DWI and DKI avoid necrosis, cystic degeneration or vascular areas, which may not be conducive to a comprehensive evaluation of tumor tissue structure. This is due to the large variation in these areas between lesions, which can interfere significantly with the parameter values. Fourth, Since the main purpose of this study was to evaluate the diagnostic efficacy of PET/MRI, there was no comparison with PET/CT, which may make this study less informative for clinical applications. Finally, this study is a limited cohort single-center analysis, which may have resulted in selection bias. In future studies, we will continue to expand the sample size and try to reduce the impact of these limitations through several methods.18F-FDG PET imaging in PET/MRI are all effective methods to distinguish benign from malignant SPLs and are also helpful in the diagnosis of pathological grading of malignant SPLs. The combination of the independent predictors SUVmax and MK had higher diagnostic efficacy than MD in differentiating benign from malignant SPLs.DWI, DKI, and The original contribution presented in this study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author.The studies involving human participants were reviewed and approved by ethics committee of Henan Provincial People\u2019s Hospital. The patients/participants provided their written informed consent to participate in this study.MW: Conceptualization, Funding acquisition, Methodology, Writing-Reviewing and Editing, Validation. JC: Conceptualization, Methodology, Writing-Reviewing and Editing. ZL: Data curation, Methodology, Formal analysis, Writing-Original draft preparation. NM: Methodology, Writing- Reviewing and Editing. YL: Formal analysis, Investigation. HJ: Formal analysis, Writing-Original draft preparation. ZH: Methodology, Investigation. TF: Investigation. PF: Investigation. FF: Project administration, Investigation, Funding acquisition. YB: Project administration, Investigation. WW: Project administration, Investigation. YY: Software, Writing-Reviewing and Editing. JY: Software. All authors contributed to the article and approved the submitted version.This work was supported by the National Key R&D Program of China [grant number 2017YFE0103600], the National Natural Science Foundation of China [grant number 81720108021 and 31470047]; and the Zhengzhou Collaborative Innovation Major Project [grant number 20XTZX05015]; and the Henan provincial science and technology research projects [grant number 212102310689], and Key Project of Henan Province Medical Science and Technology Project (LHGJ20210001).JY and YY are employees of United Imaging Healthcare (UIH).The remaining authors declare no relationships with any companies whose products or services may be related to the subject matter of the article.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "The present manuscript aims to describe an international, electronic-based, user-friendly and interoperable patient registry for monogenic autoinflammatory diseases (mAIDs), developed in the contest of the Autoinflammatory Diseases Alliance (AIDA) Network.This is an electronic platform, based on the Research Electronic Data Capture (REDCap) tool, used for real-world data collection of demographics, clinical, laboratory, instrumental and socioeconomic data of mAIDs patients. The instrument has flexibility, may change over time based on new scientific acquisitions, and communicate potentially with other similar registries; security, data quality and data governance are corner stones of the platform.AIDA project will share knowledge and expertise on mAIDs. Since its start, 118 centers from 24 countries and 4 continents have joined the AIDA project. Fifty-nine centers have already obtained the approval from their local Ethics Committees. Currently, the platform counts 337 users . The Registry collects baseline and follow-up data using 3,748 fields organized into 21 instruments, which include demographics, patient history, symptoms, trigger/risk factors, therapies, and healthcare information for mAIDs patients.https://clinicaltrials.gov.The AIDA mAIDs Registry, acts both as a research tool for future collaborative real-life studies on mAIDs and as a service to connect all the figures called to participate. On this basis, the registry is expected to play a pivotal role in generating new scientific evidence on this group of rare diseases, substantially improving the management of patients, and optimizing the impact on the healthcare system. NCT 05200715 available at Constitutive overactivation of these pathways leads to increased release of monocyte- and neutrophil-derived cytokines, such as interleukin-1\u03b2, tumor necrosis factor \u03b1 and type 1 interferon, which results clinically in periodic fevers and a variety of unprovoked inflammatory symptoms affecting any organ or system , 3. Givean organized system that uses observational study methods to collect uniform data to evaluate specified outcomes for a population defined by a particular disease, condition, or exposure, and that serves one or more predetermined scientific, clinical, or policy purposes\u201d the EC seeks to harmonize registry data to ensure interoperability, standardization, and data comparability .The present manuscript aims to describe an international, electronic-based, user-friendly and interoperable patient registry for mAIDs, the Autoinflammatory Diseases Alliance Registry of Monogenic Autoinflammatory Diseases (AIDA mAIDs), developed in the context of the AIDA Network program and administrative support are provided by the University of Siena.The AIDA mAIDs is an electronic-based, non-population based, physician-driven, clinical/genetic research registry .The overarching scope of the AIDA registry for mAIDs is declined into the following general aims: (i) to share knowledge and expertise on mAIDs by linking key referral centers for these rare diseases at an international level; (ii) to raise awareness among physicians and the general population about mAIDs, improving early diagnosis; (iii) to promote future multicenter studies based on a critical volume of data from patients with mAIDs.The AIDA mAIDs-based studies will pursue the following specific objectives:to depict the clinical phenotype of newly identified mAIDs and broaden the phenotypic spectrum of well-established nosological entities;to describe genotype-phenotype correlations;to describe clinical manifestations based on patients' ethnicity;to identify age- and gender-related factors that may affect disease onset and progression;to outline the long-term prognosis for these diseases and identify potential prognostic factors for negative outcomes;to evaluate response to different therapeutic strategies and safety of drugs;to recognize the possible impact of mAIDs on fertility and course of pregnancy;to estimate the socio-economic burden of mAIDs.Further objectives may be added over time thanks to the flexible modular infrastructure of the registry.All hereditary autoinflammatory diseases (monogenic forms) will be included in the registry. As new diseases are identified in this group, the registry shall be updated accordingly given its flexible framework. The list of genes/diseases currently included in the registry is given in Patients will be enrolled on the AIDA registry if they meet all the following criteria:https://aidanetwork.org/en/clinical-sites).Subjects diagnosed and/or treated at the AIDA network partner centers (the full list is available at Diagnosis of mAIDs based on one of the following scenarios:a) the presence of a confirmatoryb) the presence of a not confirmatory (see text footnote 1) genotype AND at least 2 among the clinical items included in the Eurofever classification criteria for CAPS, FMF or TRAPS;c) fulfillment of the Tel Hashomer or the Yalcinkaya criteria for FMF, irrespectively of the genotype , 15;d) fulfillment of the Kuemmerle-Deschner criteria for CAPS, irrespectively of the genotype ;e) the presence of a clinical picture consistent with MKD or adenosine deaminase (ADA)2 deficiency AND a positive biomarker test for DADA2 (ADA2 enzyme activity) or MKD AND an inconclusive genotypef) the presence of a clinical picture consistent with Blau syndrome, pyogenic arthritis-pyoderma gangrenosum-acne (PAPA) syndrome, A20 haploinsufficiency, ADA2 deficiency or pyrin-associated autoinflammation with neutrophilic dermatosis (PAAND) syndrome AND the detection of a confirmatory or consistent genotype (see text footnote 2) ;g) the presence of a clinical picture consistent with any monogenic autoinflammatory disease covered by the registry other than FMF, MKD, TRAPS, CAPS, Blau syndrome, PAPA syndrome, A20 haploinsufficiency, DADA2 and PAAND syndrome AND a co3. Willing of the subject to join the project.Subjects carrying benign variants, likely benign variants (based on the INFEVERS classification) or no variants in genes known to be responsible for mAIDs, except for the scenarios described in 2c and 2d, are excluded from the registry . SpecifiThe data sources for the registry are extracted from (i) hospital clinical charts, (ii) laboratory reports, (iii) genetic laboratory reports, (iv) instrumental exams reports, (v) patient reported outcomes. The registry system is designed to capture both retrospective and longitudinal data.To minimize recall bias for self- or proxy-reported information in the retrospective section, participants are notified before sensitive information is asked and they are allowed to provide secondary sources of information to update or validate previously collected data. As for the prospective section of the registry, a streamlined approach through real-time ad-hoc updates to the records during routine follow-up visits is suggested. Recruiting centers are advised to enter at least one follow-up record per year or when therapeutic changes are made.The system includes mandatory and non-mandatory fields. However, it allows editing and completing any previously unanswered fields at the user's convenience. User-friendly data collecting tools are in place, such as automatic calculation fields, calendar fields, branching logics, electronic joint count homunculus and integrated Online Mendelian Inheritance in Man (OMIM), HUGO Gene Nomenclature Committee (HGNC) and International Classification of Diseases (ICD) 10 databases. Direct explanations or Internet addresses referring to external resources useful to the interpretation of specific fields are provided when required [i.e., INFEVERS classification of gene variants , diagnosThe target population includes subjects affected by mAIDs. No specific demographic, geographic, clinical, or genetic determinants are foreseen.https://aidanetwork.org/en/clinical-sites.The AIDA mAIDs registry is a non-population-based registry. The geographic coverage is the catchment area of the clinical centers affiliated to the AIDA Network. The countries with at least one AIDA partner center are shown in The instruments constituting the registry investigate the following sets of variables:demographicsconsentsdiagnostic data and family historygeneral genetic information:- gene mutationsfeatures of inflammatory attacks:- at disease onset- between disease onset and diagnosis- between diagnosis and the time of enrolmentclinical diagnostic and classification criterialaboratory datacardiovascular riskpast and current treatments:- treatment with corticosteroids as main therapy- treatment with colchicine- treatment with conventional disease-modifying anti-rheumatic drugs- treatment with small molecules- treatment with biotechnological agentsfertility and pregnancy- disease course and treatment during pregnanciesfollow-up visits: clinical manifestations and treatmentDeath of the patient (to open ONLY in case of patient's death)common data elements (CDE) are included in the registry, according to the EPIRARE set of CDE for the European RDR platform (The following platform : patientThe ELSI and privacy expertise are provided by the University of Siena. The AIDA project has been firstly approved by the Tuscany Region Ethics Committee - South-East (C.E.A.V.S.E.) area on 24/06/2019 (Ref. N. 14951). The last amendment to the protocol was approved on 02/05/2022. The approval of the protocol should be provided by local Ethics Committee for each of the Centers joining AIDA, whenever required by local regulations. The approval is essential for data collection, but does not affect the participation to the other branches of the AIDA project, such as AIDA Academy or AIDA for patients.The registry has been developed in accordance with the World Medical Association (WMA) Helsinki declaration 2013 ensuringTo be eligible for inclusion in the AIDA registry for mAIDs, patients have to provide written opt-in consent. Patients receive from the investigator appropriate information about registry objectives, the type of information collected, how data will be used, the governance and data access rules for third parties and how to withdraw consent at any time.https://aidanetwork.org/en/). Established in 2019, the program aims to move beyond the isolation of reference centers for rare autoinflammatory diseases and autoimmune ocular diseases, facilitating the collection and exchange of clinical data, conduction of multicenter studies and dissemination of scientific knowledge at an international level.The AIDA registry for mAIDs was developed as a specific action of the AIDA Network program , researchers, the European Reference Network (ERN) RITA.https://eu-rd-platform.jrc.ec.europa.eu/erdridor/home) and is committed to promote the interoperability with the other ERN RITA registries and potentially with the other ERNs, within the context of the MeRITA (Metadata registry for the ERN RITA) project .https://projectredcap.org) a secure web application designed to support data capture for research studies. REDCap provides (i) an intuitive interface for validated data capture; (ii) audit trails for tracking data manipulation and export procedures; (iii) automated export procedures for seamless data downloads to common statistical packages; and (iv) procedures for data integration and interoperability with external sources assess, deploy and manage multiple aligned REDCap framework instances and project registries; (ii) efficiently share and deploy standard resources, such as medical data models and ontologies, that already exist or that are newly released by international framework initiatives (such as ERDRI-JRC and ERNs), integrating them into the network of project registries and data collection instruments; (iii) ensure compliance with privacy and security requirements throughout the project partners; and, (iv) guarantee global sharing and FAIR access to data.http://support@aidanetwork.org). The REDCap system allows quality data control at the moment of the data entry by users. In addition, internal data quality audits are periodically performed by the registry staff at the time of sample data extraction or when significant sources of error are identified. The quality control at the time of data extraction is aimed at the control of duplicate records from different investigator sites or logical inconsistencies and range errors on individual data batches sent by each site. When range errors or logical inconsistencies are found, a query is sent to the correspondent principal investigator in order to check the data source. When duplicate records are identified, the record management is dependent on the type of duplication and objective of analysis.The principal investigators of each site are responsible for the validation of data of the corresponding records. They will be also responsible for the accuracy of the information accrued, with the Principal Investigator required to supervise the goodness of overall data. Each Principal Investigator may analyze data gathered in his/her center. Site opening visits are scheduled virtually for each new partner center in order to train the investigators in the correct use of the registry. Further assistance from the AIDA team may be asked by the investigators by email from 35 centers in 11 countries have been enrolled in the registry. Enrolling countries listed in alphabetical order are the following: Algeria, Belgium, Brazil, Egypt, Greece, Iran, Italy, Poland, Romania, Spain, Turkey. Patients' country of birth are Algeria (n = 23), Armenia (n = 1), Belgium (n = 4), Brazil (n = 1), China (n = 1), Egypt (n = 58), Greece (n = 6), Iran (n = 3), Italy (n = 195), Lebanon (n = 1), Morocco (n = 2), Palestine (n = 1), Poland (n = 27), Romania (n = 4), Spain (n = 2), Turkey (n = 78), Ukraine (n = 1), missing (n = 10). Mean age at the time of enrolment is 33.9 \u00b1 16.7 years (range 0\u201375.9). A positive family history for the same mAID is recorded for 162 subjects (40.9%). Proband's parents are consanguineous in 40 cases (10.2%).At last evaluation , 418 subjects are transnational and devoted exclusively to AIDs (in two cases to a single specific disorder); four other registries are transnational and include both autoinflammatory diseases and primary immunodeficiencies and/or autoimmune diseases. The remaining registries collect regional or national data about rare immune diseases, including autoinflammatory diseases, with variable specificity . TherefoIn the scene of European registries for mAIDs, the AIDA registry stands out for its disease-specificity, richness in details, flexibility, complex branching logics allowing smart and time-saving data collection, and wide geographic and demographic coverage. With specific regard to the latter, the choice of including subjects of any age naturally follows from the well-established evidence that mAIDs may start from the very first hours of life to late adulthood, and also that adults with pediatric-onset disease may obtain the correct diagnosis with a delay of several decades. The inclusion of adult patients along with children is an asset to this registry, allowing the design of comparative and longitudinal studies, research focused on childhood-to-adult transition and adult-specific issues not yet explored, such as pregnancy, fertility, adult vaccination, comorbidities, long-term disease-related damage, adult-specific outcome measures and PROMs, workability and further socio-economic issues. With this respect, the AIDA actions will be aligned to the research agenda set by the EULAR/ACR points to consider for diagnosis, management and monitoring of the IL-1 mediated AIDs and autoinflammatory type I interferonopathies , 27, alshttps://eu-rd-platform.jrc.ec.europa.eu/erdridor/) and will adopt the new SPIDER tool in the future, with the aim of facilitating the pseudonymization, linkage and transfer of encrypted pseudonymized data among European rare disease registries.The registry has been conceived as a flexible and modular tool able to capture the evolving landscape of this field of research. Whensoever new theoretical or practical knowledge is generated, the system enables agile updating of data collection tools. As an example, new modules may be added to include newly identified genes, new treatments that become available or to address to future unmet needs with new research objectives. Moreover, direct queries to the investigators can address specific gaps in the data collection. The AIDA mAIDs registry is inspired by the principles of FAIRness and is committed to adopt the instruments that the EC suggests for the development of registry platforms. Data are standardized by the use of shared libraries such as the ICD-10 and the OMIM classification; the EPIRARE set of CDE for the European RDR platform has been employed when possible . The regvia user-facing surveys and the MyCap application (https://projectmycap.org/), which leverages REDCap, ResearchKit, and ResearchStack tools to capture patient reported outcomes via mobile devices. Later, the tool synchronizes the results back to the registry project. The integration of data from the \u201cAIDA for patients\u201d action into the AIDA mAIDs registry highlights the huge potentiality of this instrument. Actually, data obtained through the surveys proposed by \u201cAIDA for patients\u201d will complement the registry with patient-reported data about quality of life, fatigue, the socio-economic burden of the disease, psychological health, experience of the healthcare pathway and beliefs about medicines. This will allow four-hands studies with the active participation of both patients and their physicians. With this regard, a pilot project co-designed with the patient's association S.I.M.B.A. has been already launched by the \u201cAIDA for patients\u201d action for Italian patients with Beh\u00e7et's disease. A similar experience may be reproduced also in the context of mAIDs and other multifactorial AIDs, with the collaboration of local patient associations (https://aidanetwork.org/en/magazine/aida-for-patients-is-ready-for-launch).On the other hand, the registry platform supports data capture AIDA Academy action). Of note, the program is endorsed by a growing number of patient associations, whose active involvement enhances a multiplier effect, by giving resonance to both AIDA Network program and AIDA mAIDs registry.As for the long-term sustainability of the registry, it seems relevant to highlight the lively interest raising from a growing number of international partners both in and outside European borders. Moreover, the AIDA Network program strategic communication and dissemination activities are equally important to the registry promotion. The program also includes the provision of high-level specialized education in the field of AID, through web-based seminars, face-to-face meetings, and a permanent education archive in collaboration with a renowned international faculty area on 24/06/2019 (Ref. N. 14951). Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.CG wrote the first draft of the manuscript. AV conceived and designed the study and revised the draft of the manuscript. DR revised the draft of the manuscript. LC conceived and designed the study and accounts for AIDA Registries Coordinator. AB is the bioengineer involved in the technical management of the platform and registries. AT, GR, EA, EW-S, DA-I, GC, LI, MCM, MC, FLT, GL, EV, AP, AS, AI, PPS, AM, MF, BO, DO-B, PP, GE, FS, FCa, MP, JH-R, RP, MA, RNu, AO, ED, PS, PR, FLG, HK, JS, MAH, GM, KJ-R, RS-H, MR, FR, FCi, FI, FD, MF, KL, TG, FF, VSa, DY, VC, MTH, RD, AK, and GK were involved in data recruitment in the Registry dedicated to patients with mAIDs. MT, IA, AL, VM, LD, LB, SGe, FCr, VP, AA-MAM, RNa, MD, CBC, SGr, MM, ID, VSp, HAMG, IV, AR, AF, MAM, BF, GT, and CF were included in the authorship as investigators from the top contributor centers for any of the other seven AIDA Registries. ML was included as leading AIDA expert in the field of mAIDs. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Motivation and anxiety, which are classified both as psychological and as affective factors, have been shown to play important roles in second-/foreign-language (SL/FL) learning at all levels. Specifically, students with higher motivation tend to study their target languages (TLs) more effectively, and those with high anxiety, less effectively. Additionally, scholars agree that SL/FL motivation and anxiety (1) are closely related to each other; (2) interact with other variables to affect SL/FL learning; and (3) are dynamic, and thus that their effects on SL/FL learning are also dynamic. Nevertheless, in light of the complex nature of language learning and the huge diversity of SL/FL learning populations and contexts, both motivation and anxiety remain under-researched. Conducted in diverse contexts, the 15 articles making up this special issue expand research on anxiety, motivation, and their relations with SL/FL learning both theoretically and methodologically.Gao; Wang and Xue) of research on anxiety and motivation, respectively. Wang and Xue focus on how the expectancy-value motivational model impacts academic motivation, engagement, participation in educational tasks, and academic performance. Gao classifies existing FL-anxiety scales into three types: test-based, measuring speaking anxiety; classroom-based, measuring speaking anxiety; and activity-based, also measuring speaking anxiety. She also introduces Classical Testing Theory and Rasch measurement as two major statistical paradigms for guaranteeing the reliability of these scales. As well as summarizing the emerging themes of the relevant research, the author discusses the dynamic approach to interpreting the interrelationship of anxiety, language performance, and other factors involved in language learning, and highlights possible directions for future anxiety-related research.Of these 15 articles, two are reviews ; four, quantitative methods ; one, qualitative methods (Lu and Yoon); and the remaining three, experimental designs . They examine various aspects of anxiety and motivation including theory development, strategies, measurements, effects, and sources.The remaining 13 papers all report on empirical research: five using mixed methods (Yan and Liang's contribution investigates the effects of English-Chinese interpretation-classroom foreign language anxiety (ICFLA) on interpretation learning and dependency distance (DD) among 49 undergraduate and graduate students in Hong Kong. They report a significant negative correlation between ICFLA levels and consecutive interpretation achievement scores. ICFLA was also negatively correlated with DD in consecutive interpretations. Rasool et al.'s article explores levels of and reasons for writing anxiety, and gender influence on anxiety levels, among 72 pre-service FL teachers in Pakistan. It reports that most of the participants experienced medium to high writing-anxiety levels, without gender differences, that were accounted for by linguistic challenges, fear of negative judgment, lack of self-confidence, and bad prior experiences. Lin's contribution uses interview and survey data from 243 Chinese students of L3 French at a university in the UK to explore the relationship between their L3 anxiety and their self-efficacy, which emerged as negative. It also shows that grammatical and pronunciation similarities between English\u2014the participants' L2\u2014and French positively decreased these students' anxiety levels.Three of the papers focus on anxiety. Ren and Abhakorn's explores the psychological and cognitive factors behind college students' loss of motivation to learn English in universities in China, and the interrelationships of those factors. Specifically, they constructed a shopping-cart model based on the results of 23 interviews and used structural equation modeling to test it on questionnaire data from 286 demotivated students. This revealed three distinct pathways whereby the respondents were demotivated: (1) a large discrepancy between their actual and required positioning of English learning, (2) a low required positioning of English learning, and (3) a low value of English learning in students' minds. He et al. also studied Chinese university students, collecting interview and survey data from 79 of them and investigating their perceptions and practices of rubric use throughout a task process. Their results highlight the important roles of trait motivation and task motivation in the effectiveness of rubric use during assessment. Greenwald et al.'s contribution, meanwhile, looks at intrinsic and extrinsic motivation among 851 monolingual and 196 bilingual children in the United States, and suggests that among the latter group, these two types of motivation are not antagonistic\u2014unlike with their monolingual counterparts.Three further contributions focus on motivation. Dong, Liu et al.'s based on a questionnaire survey of 280 Chinese high school students, explores the relations among FL classroom anxiety (FLCA), enjoyment (FLE), and expectancy-value motivation, as well as how effectively these three variables predict students' self-rated FL proficiency. It reports that (1) the students' FLE was significantly and positively correlated with all dimensions of expectancy-value motivation, whereas their FLCA and expectancy-value motivation demonstrated a complex correlation pattern; and (2) expectancy beliefs, intrinsic value, private enjoyment of FL learning and anxiety arising from fear of negative evaluation jointly and significantly predicted the students' self-rated FL proficiency. Dong, Jamal Mohammed et al. report on their exploration, via a pre- and post-test experimental design, of the effects of three instructional modes\u2014computer-assisted language learning (CALL), mobile-assisted language learning (MALL), and face-to-face (FTF) learning\u2014on Iranian EFL learners' motivation, anxiety and self-efficacy. Specifically, they randomly assigned 30 such learners to each of three classes, each comprising 25 1-h sessions, and found that the experimental groups' motivation, anxiety, and self-efficacy were positively affected by CALL-based and MALL-based instruction, though there was no statistically significant difference between the CALL and MALL groups in this regard. Zhang and Dong's paper examines how 230 Chinese college students' motivational-regulation strategies affected their proximal and distal L2 writing-achievement emotions , and tested for possible interactive effects of such strategies and self-regulated learning strategies on the same two emotions. They report that all the motivational-regulation strategies that they studied directly predicted both proximal and distal writing enjoyment, but that only a performance-oriented one predicted proximal or distal writing anxiety. Another key finding is that a social-behavior learning strategy counteracted the high proximal anxiety caused by heavy use of the performance self-talk motivational-regulation strategy. Moreover, it highlights motivational-regulation strategies as stable predictors of both proximal and distal writing wellbeing. Wu et al.'s contribution, based on a sample of 223 students of the Top-Notch Students of Basic Disciplines Training Program in a top Chinese university, examines English-use anxiety (EUA), motivation, self-efficacy, and use of English, along with these variables' predictive effects. Their findings indicate that (1) in general, EUA and language-learning orientation were significantly and negatively correlated, and significantly but positively correlated with the other measured variables; (2) the participants' EUA and intrinsic-motivation knowledge significantly predicted their English achievement; and (3) their use of English and self-efficacy mediated the effects of EUA and language-learning orientations on their English achievement.Four studies examined the interaction of anxiety and motivation. Lu and Yoon's paper reports on how they used interview, textual and documentary data to examine the influence of power relations on the research practices of six EFL academics at a Chinese university, as well as the same individuals' coping strategies. They conclude that, even though their participants were driven to engage in research by a combination of intrinsic and extrinsic motivations, their research endeavors were undermined by the marginalized status of EFL researchers from non-elite universities, as imposed by the Chinese academic circle. Even so, they exerted their agency via micropolitical literacy and tried to seek ways out of their unfavorable academic culture. Izadpanah's contribution involves a pre- and post-test experimental study of 354 high-school students, aimed at ascertaining the impact of flipped teaching (FT) on EFL students' academic resilience (AR), self-directed learning (SDL), and learner autonomy (LA). It shows that FT significantly affected AR, SDL, and LA, and that the mean scores of EFL students' AR, SDL, and LA were higher through FT. Last but not least, Rezaee and Seyri's piece examines boredom as experienced by 84 online students of English for academic purposes and the success of an autonomy-oriented intervention program aimed at alleviating such boredom.The remaining three studies focused on other issues related to motivation. Collectively and individually, these studies shed considerable light on teaching and learning, both theoretically and empirically. Nevertheless, more research on anxiety and motivation is still warranted. For example, more longitudinal studies are needed to document changes in people's anxiety and motivation, and such changes' effects on SL/FL learning. Likewise, there needs to be more research on both technological and non-technological strategies for intervening to reduce anxiety and increase motivation in the context of such; and anxiety and motivation connected with the learning of SLs/FLs other than English remain under-researched.ML drafted the editorial. C-HL and YZ revised and polished the editorial. All authors contributed to the article and approved the submitted version."} +{"text": "Children's environments - especially relationships with caregivers - sculpt not only developing brains but also multiple bio-behavioral systems that influence long-term cognitive and socioemotional outcomes, including the ability to empathize with others and interact in prosocial and peaceful ways. This speaks to the importance of investing resources in effective and timely programs that work to enhance early childhood development (ECD) and, by extension, reach communities at-scale. Given the limited resources currently devoted to ECD services, and the devastating impact of COVID-19 on children and communities, there is a clear need to spur government leaders and policymakers to further invest in ECD and related issues including gender and racial equity. This essay offers concrete examples of scholarly paradigms and leadership efforts that focus on child development to build a peaceful, equitable, just, and sustainable world. As scholars and practitioners, we need to continue to design, implement, assess, and revise high-quality child development programs that generate much-needed evidence for policy and programmatic changes. We must also invest in global partnerships to foster the next generation of scholars, practitioners, and advocates dedicated to advance our understanding of the bio-behavioral systems that underlie love, sociality, and peace across generations. Especially where supported by structural interventions, ECD programs can help create more peaceful, just, and socially equitable societies. \u2022Science has documented the lasting long-term benefits of Early Childhood Development programs from pre-gestation onwards.\u2022Sadly, many children and their families, especially in conflict-affected areas, are exposed to toxic environments.\u2022In addition, too many children are now being exposed violence online and, in their homes, schools, and communities.\u2022There is a need for government leaders and policymakers to invest in ECD programs that promote gender and racial equity.\u2022We also need to partner with children, youth, and their families to build more a peaceful and socially cohesive world. Difference is an accident of birth and therefore should never be a source of hatred or conflict. The answer to difference is to respect it. Therein lies a most fundamental principle of peace \u2013 respect for diversity.John HumePeace is integral to human existence. In everything we do, in everything we say, in every thought we have, there is a place for peace. Early childhood affords a window of opportunity to instill what each individual needs to become an agent for peace and nonviolence.H.E. Ambassador Anwarul K. Chowdhury1via nurture\u201d opportunity to become an agent for peace and nonviolence\u201d. In addition, the scientific and programmatic evidence in support of ECD programs as a pathway to peace was thoroughly reviewed by a large group of interdisciplinary experts during the meetings, such as the 15th Ernst Str\u00fcngmann Forum [We turn to leadership efforts \u2013 such as those of ECPC and ECDAN - that focus on comprehensive child development and global partnership initiatives to build a more peaceful, equitable, and sustainable world. The ECPC was founded in the belief that children and families can be agents of change to promote love, empathy, and peace . Its forf peace\u2019 \u2013 under nn Forum ,62.https://ecdpeace.org/work-content/read-ecpc-global-call-action-reponse-covid-19). In leveraging early childhood development as a pathway to empathy and sociality, ECPC team members have worked towards four dimensions of peace: 1) as an outcome defined by the cessation of violence, 2) a process to build non-violent social bonds, 3) a disposition predicated on prosocial actions and respect for human dignity, and 4) a culture that fosters a sense of global citizenship.The mission of the ECPC is to create an inclusive movement for peace, social cohesion, and social justice. Its three main goals, presented in https://www.ecdan.org/). ECDAN works in close alignment with seven regional networks, each moving ahead to create, test, refine, and bring to scale high-quality ECD services [https://bernardvanleer.org/ecm-article/2018/supporting-refugee-internally-displaced-and-marginalized-host-community-parents-in-the-arab-region/). Interventions will continue to improve with the growth of developmental science, but in most instances, ECD programs must build on existing delivery platforms to enhance their feasibility and ensure that they can be successfully scaled-up [https://www.ecdan.org/index.html).The ECDAN was launched in 2016 by the World Bank and UNICEF and more than 80 organizations across the globe, in an effort to catalyze collective action on behalf of young children and their families, facilitating knowledge exchange and coordinating advocacy for investing in quality services and the Sou da Paz Institute in Brazil (https://soudapaz.org). Another example is the Arab-German Young Academy of Sciences and Humanities . AGYA was established as a collaborative effort between researchers in 22 countries in the Arab world and Germany. AGYA's initiatives focus specifically on education at the interface of science and society. In AGYA, early-career researchers seek to jointly address the challenge of access to quality ECD services by building on mutual trust and knowledge exchange.There are also multiple examples of local initiatives and organizations that support young people's involvement in peacebuilding such as the Africa Youth for Peace and Development in the United Kingdom through the LINKS initiative . These include programs in Columbia, Palestine, Egypt, Mali, Kyrgyzstan, Tajikistan, Timor-Leste, and Vietnam . We detaNorthern Ireland. The Good Friday Agreement, led by the late John Hume, has set the stage for ECD programs, focused on peacebuilding in Northern Ireland. The Media Initiative \u201cToo Young to Notice?\u201d was developed following ground-breaking research showing that children as young as 3 years of age were developing negative and prejudicial attitudes to those who were different from them [https://www.early-years.org/).rom them . Severalrom them ,72. The Turkey. As a result of forced displacement in the wake of the Syrian conflict that began over a decade ago, there are an estimated 3.6 million Syrian refugees now living in Turkey. While 1.7 million of those registered are children, 860,000 of them are school-aged and have limited access to schools. The European Union, with additional support of the Turkish Ministry of National Education, has been actively involved in the integration of Syrian children into the Turkish Education System. Its program includes Turkish and Arabic language courses, as well as catch-up education and remedial/support classes, and transportation services for students [https://www.acev.org/wp-content/uploads/2019/10/3.Syrian-Children.CaseStudy.pdf). This initiative is closely aligned with ACEV's Preschool Education Program (PEP), implemented since 2003 in areas in Turkey where access to early childhood education rates is low [students . In addis is low ,75. As is is low .Jordan. A number of organizations are refining ECD programs that work directly with families to help alleviate profound stress and build prosocial behavior and resilience. In Jordan, a consortium of local and international scholars worked with Mercy Corps and Syrian/Jordanian communities to evaluate a structured psychosocial intervention implemented with refugee and war-affected youth [Terror and Hope: The Science of Resilience, to exemplify how science, fueled by compassion, can work to break the impacts of violence - and foster constructive dialogue between youth, families, researchers, practitioners, and policymakers (see: https://www.ronbourkefilms.com/terror-and-hope).ed youth ,78. The ed youth . The prohttps://welovereading.org/). It is a practical and sustainable grassroots approach to stimulate learning, using storybooks to foster the love of reading. Being volunteer-driven, it empowers people in marginalized communities to become social entrepreneurs and to co-develop books for children. The community-based approach is vital and fostered through interpersonal engagement. This initiative has won multiple international awards for social innovation and refugee education, and has now spread, from a local solution, to as many as 63 countries globally. Its impact on child development outcomes is now being rigorously evaluated\u00a0[Many programs and interventions have been designed, evaluated and led by people from conflict-affected regions. One noteworthy example is We Love Reading (WLR), an innovative approach developed in Jordan, by Dr. Rana Dajani, to have adult volunteers read aloud to groups of children in their local community . These aspects of research, policy, and practice synergistically affect the effectiveness and longevity of specific programs; they are cross-cutting determinants of impactful ECD programs in the context of social transformation.Both examples from Jordan highlight critical components of successful programs, namely strong partnerships, cultural engagement, and structural competency. Strong partnerships work to bring together local, regional, and international actors in ways that foster pathways to health, development, and social inclusion ,113. CulOf course, presenting examples of successful ECD programs is only a first step. We do not make an argument that the primary responsibility for positive development and larger societal outcomes is at the individual-level, given that children are embedded within social, economic, and political structures . On the 6Evidence-based recommendations, regarding how best to strengthen national policies and programs as well as frame the holistic needs of the developing child, have been articulated in The Lancet ECD series . In whatExpand political will andAction item 1: funding through advocacy for ECD-related SDGs. Under the SDG umbrella, investing in ECD is not only an aim in itself, but it is also a requisite for achieving other SDGs. For example, SDG 4.2 calls for universal access to quality ECD, care, and pre-primary education, and provides unprecedented opportunity to scale-up ECD services. Convincing government leaders and policymakers to take action at present, during the ongoing pandemic and the accompanying political and economic challenges, is an ambitious goal. That said, the data concerning the long-term value of delivering high quality ECD services are compelling [mpelling ,80. An ehttps://ecdpeace.org/work-content/theory-change-early-childhood-development-sustainable-development). Engaging the media, as with the documentary Terror and Hope: The Science of Resilience, is another important opportunity to reach wider audiences, in ways that can convince citizens and policymakers that science-based, high-quality programs are worth the investment. In addition, in order to ensure sustainability and scalability, partnerships and communication mechanisms to link civil society and social entrepreneurs are critical. Applied research studies have increasingly explored advocacy strategies to influence early childhood education systems, for example in countries like Jordan [There is certainly a need to foster strong, sustained engagement with local communities. One way is to co-produce and disseminate documents in local languages in order to convey scientific findings from global ECD programs. Developing a compelling Theory of Change is another step in the right direction .hildhood . The enahttps://www.covid19parenting.com/home). Formats include tip sheets, comic strips, radio sketches, TV broadcasts, public service announcements via loudspeakers and community radio, community worker templates, webinars, social media messaging, interactive text messages, online parent support groups, phone-based counselling, and theme songs. Parenting intervention content can also be delivered through mobile applications, texting, and chat services . Comparable efforts are underway around the world including in the Middle East and North Africa (MENA) region, thanks to the efforts of the Arab Resource Collective (ARC) (see: https://mawared.org/en/).In the context of the ongoing pandemic, ECD practitioners and scholars have launched interactive web-based services for parents that have been widely disseminated has been endorsed by the United Nations Statistical commission as the tool to measure SDG 4.2.1 and is the first ever population-based tool for measuring young children's development. A continued challenge of the field includes the harmonization of global measurement frameworks and the extent to which these are contextualized across geographies. It is imperative to include the participation of local communities into how those frameworks are formulated, implemented, and ultimately applied to strengthen service provision and guarantee the rights of children. One of these efforts emerged in response to the need to measure the progress of Target 4.2 of the SDGs (see: http://www.ecdmeasure.org/what-is-melqo/). Global multi sectoral and inter-disciplinary partnerships have operationalized the adaptation and application of these frameworks in Colombia, among other countries [he field . The recountries . It is i7We clearly need to support the next generation of youth as partners in decision-making and in the peacebuilding processes, as stipulated in the United Nations resolution (UNSCR 2205) on Youth, Peace and Security, adopted by the United Nations Security Council in 2015 . The worThe current reality, however, is that young people are still largely sidelined. Young people, especially young girls, in many regions of the world continue to be socially, economically, and politically excluded. The recent WHO\u2013UNICEF\u2013Lancet Commission identified several barriers to their engagement including traditional hierarchical governance and adultism . PoliticPakistan. In 2015, colleagues at The Edward Zigler Center in Child Development and Social Policy at Yale University partnered with researchers from Harvard University and Aga Khan Universities to develop, implement, and scale-up one of the few robustly evaluated youth-led ECD programs in LMICs. The program is entitled, Youth Leaders for Early Childhood Assuring Children are Prepared for School (LEAPS). LEAPS is a set of cross-generational programs that aim to train, mentor, and empower youth to promote the demand for quality ECD programming in their communities [munities . LEAPS imunities . Given tColombia. Working with local leaders and academic partners, the LEAPS implementing team is currently in the process of rolling out a study to assess: (a) the governmental support and assessment of governance and institutional arrangements for program sustainability and scale-up; (b) the attributes of youth buy-in for program participation and assessment of the benefits of the program for youth professional development in the Colombian context; (c) the nature of community and family perceptions and level of acceptability of youth-led programming for early childhood; and (d) the processes to establish academic partnerships and mentoring for sustained program implementation and evaluation. The initial plan is to conduct a qualitative study to characterize the above areas, as a first step to determine key attributes of the program's adaptation. The data will then be utilized as a key set of preliminary findings to rollout a culturally adapted and contextualized youth led ECD program in Colombia.Middle East. The Seeds of Peace program focuses on personal transformations of youth leaders to inspire wider societal change. The program begins with a month-long summer camp in the United States in the State of Maine. The campers are adolescents between 14 and 16 years of age who come from all across the globe, especially in areas affected by ongoing conflict including the Middle East. The selection process is competitive, and more than 300 campers are selected each year. For nearly 2\u00a0h each day, the campers engage each other directly in small-group dialogue sessions organized by conflict region. Together with their mentors, usually former campers, they tackle the most distressing and divisive issues defining their conflict, sharing their personal experiences, reflecting on competing narratives, and challenging each other's perspectives. Ms. Haifa Staiti, a Seeds of Peace graduate and the founder of Empathy for Peace - an NGO based in Canada (see: https://www.empathy-for-peace.org/) reported that the most transformative aspect of her experience at the Seeds of Peace camp, in the late 1990s, was the opportunity to connect with knowledgeable, nurturing, and empathetic adult counselors who provided support and care during the month-long camp. Counselor support, combined with the opportunity to connect with youth from \u201cthe other side of the conflict\u201d, helped \u201csow the seeds for new ways of thinking, seeing and being in the world that contributed to the person I grew up to become\u201d. This experience inspired her to establish the Canadian NGO, Empathy for Peace (see: https://www.seedsofpeace.org/).In considering the role of children and youth in building more peaceful communities, it is essential to reflect on the existing scientific literature concerning the transformative power of empathy, especially empathy for members of the \u201cout group\u201d as well as the impact of ongoing youth-based peacebuilding and conflict transformation programs that have been established around the world .https://www.heart-to-heart.ca), and the Whitaker Peace and Development Initiative, which operates in the United States as well as in countries in Africa and South America (see: https://www.wpdi.org). As mentioned before, there are also multiple examples of local initiatives and organizations that support young people's involvement in peacebuilding such as the Africa Youth for Peace and Development (https://africayouth.webs.com) and the Sou da Paz Institute in Brazil (https://soudapaz.org).In addition to the U.S.-based Seeds of Peace initiative, there are many similar exemplars operating globally, including the Heart-to-Heart program in Canada and in their homes, schools, communities, and societies. To break the cycle of violence and promote peaceful societies, there is a clear need for scholars, practitioners, policymakers, and other civic society leaders to partner with children, youth, and families to build more peaceful, gender equitable, socially cohesive, and resilient communities. In building back better, governments and policymakers, supported by the international community, need to prioritize investments in ECD programs and services, including initiatives that address the ongoing pandemic. However, efforts to support ECD must go hand-in-hand with initiatives to ameliorate structural injustices and institutionally entrenched inequalities that deeply impact development, including poverty and racial discrimination. Through strong coalitions and determined efforts to invest in children and the societies they live in, we can build a peaceful, equitable, just, and sustainable world and drive social change leaving no child and family behind.This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Background: Metastatic colorectal cancer (mCRC) imposes a heavy tumor burden worldwide due to limited availability of therapeutic drugs. Aflibercept, a kind of recombinant protein of the anti-vascular endothelial growth factor (VEGF) family, has been approved in clinical application among mCRC patients since 2012. A comprehensive analysis of the efficacy, safety, and cost-effectiveness of aflibercept in mCRC treatment is necessary.Objective: To evaluate the efficacy, safety, and cost-effectiveness of aflibercept for the treatment of mCRC in order to provide a decision-making reference for the selection of targeted drugs for second-line treatment of mCRC in Hong Kong, Macao, and Taiwan regions of China and the selection of new drugs for medical institutions in these regions.Methods: A systematic retrieve on databases including PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure (CNKI), Wanfang, and Weipu, as well as relevant websites and databases of health technology assessment including the National Institute of Health and Clinical Optimization, Centre for Evaluation and Communication at the University of York, and the Canadian Agency for Medicines and Health Technology, was conducted. The literature was screened according to the inclusion and exclusion criteria, and data were extracted and analyzed by two authors, while the quality of the literature was assessed.Results: Finally, we included two HTA reports, 11 systematic reviews/meta-analyses, and two cost-effectiveness studies in the rapid health technology assessment. For mCRC patients receiving second-line treatment, aflibercept combined with FOLFIRI significantly increased progression-free survival (PFS) and overall survival (OS) and the objective response rate (ORR) also improved, compared with folinic acid + fluorouracil + irinotecan (FOLFIRI). In terms of safety, mCRC patients who received aflibercept combined with FOLFIRI therapy had a higher incidence of grade 3\u20134 adverse events than those who received FOLFIRI alone, including anti-VEGF\u2013related adverse events and chemotherapy-related adverse events . In terms of cost-effectiveness, two economic studies conducted in the United Kingdom and Japan, respectively, found that compared with FOLFIRI, aflibercept combined with FOLFIRI had no cost-effectiveness advantage in mCRC patients receiving second-line treatment.Conclusion: Compared with FOLFIRI treatment, aflibercept combined with FOLFIRI for the second-line treatment of mCRC patients has better efficacy, worse safety, and is not cost-effective. More high-quality clinical studies are required for further exploration of aflibercept\u2019s clinical value. Medical institutions in Hong Kong, Macao, and Taiwan regions of China should be cautious when using or introducing aflibercept plus FOLFIRI as a mCRC treatment. Colorectal cancer, as one of the most common gastrointestinal malignancies, features high incidence, high death rate, and low cure rate and seriously threatens human health. The 2020 data showed that the incidence and mortality rate of colorectal cancer in the world, respectively, ranked third and second of all cancers, of which 1.932 million were new cases and 935,000 deaths . Patientn the clinical environment. The therapy has been approved for nearly a decade, and data on its efficacy, safety, and cost-effectiveness have been accumulating through this time. A certain number of secondary literature and economic studies on clinical efficacy and safety have been accumulated around aflibercept, which provide an evidence basis for rapid evaluation. The objective of this study is to evaluate the efficacy, safety, and cost-effectiveness of aflibercept in the treatment of mCRC in order to provide a decision-making reference for the selection of targeted drugs for second-line treatment of mCRC in Hong Kong, Macao, and Taiwan regions of China and the selection of new drugs for medical institutions in these regions.Health technology assessment (HTA) can systematically evaluate the technical characteristics, effectiveness, safety, and socioeconomic attributes of health technologies, providing decision makers of health and healthcare and medical personnel with scientific information and an evidence-based basis for the rational choice of health technologies . It takeWe included published HTA reports, systematic reviews (SR) or meta-analyses, and pharmacoeconomic studies.Patients diagnosed with mCRC were of any gender, ethnicity, onset, and origin, but all the patients should be adults. Considering the current status of antineoplastic drug research and the clinical characteristics of adverse events, for safety, a wider range of tumor patients were included for a more comprehensive assessment.The trial group consisted of aflibercept monotherapy or chemotherapy (CT), and the control group was CT with or without other positive controls or the best supportive care. Both the experimental group and the control group were second-line treatments for mCRC, with unlimited doses and courses of treatments.Efficacy measures include overall survival (OS), progression-free survival (PFS), objective response rate (ORR), complete response (CR), partial response (PR), and disease control rate (DCR). Safety indicators include the incidence of overall adverse events (AE), incidence of serious adverse events, and incidence of various types of adverse events. Economic indicator includes the incremental cost-effectiveness ratio (ICER).The exclusion criteria include (1) repeated publications, (2) literature with lack of data or inability to obtain the full text, and (3) non-Chinese and English literature.aflibercept, systematic review, Meta-analysis, economics, cost, economics, and health technology assessment as keywords in English and Chinese, respectively, with a search time frame from the date of database creation to 11 November 2021. In addition, as a supplement, a manual search of references of included studies was conducted. The search strategy for PubMed, as an example, was as We searched databases including PubMed, Embase, the Cochrane Library, CNKI, Wanfang, and Weipu, as well as the official websites and related databases including the National Institute of Health and Clinical Optimization, Centre for Evaluation and Communication at the University of York, and Canadian Agency for Medicines and Health Technology, and included HTA reports. SR or meta-analysis and pharmacoeconomic studies on aflibercept for metastatic colorectal cancer were searched in full text with After the literature was deduplicated, two researchers (Pu Ge and Xiaonan Wang) screened and cross-checked by reading the title, abstract, and full text according to the inclusion and exclusion criteria, and if there was any disagreement, they would negotiate with the third researcher (Xiao Han).The basic data were independently extracted by two researchers according to the pre-designed data extraction table, including first author, publication year, intervention/control measures, and outcome indicators. If the included literature was incomplete, we would contact the original author to obtain it.Here, two investigators used different tools to evaluate the quality of all included literature. For HTA, the HTA checklist was usedBecause of the high heterogeneity of study types, this study used descriptive methods to assess outcomes. Qualitative profiling methods were used to classify, compare, and analyze the results of the included studies according to the study design, patient population, and intervention/control measures of each study.The main characteristics of the included HTA reports, SR/meta-analyses, and economics studies are reported in The quality evaluation results of HTA are shown in The quality evaluation results of the SR/Meta-analysis are shown in The quality evaluation results of economic research are shown in The HTA report from theA total of two HTA reports and fourp < 0.001). The median PFS of the experimental group was 6.90\u00a0months (95% CI 6.51\u20137.2 months) and that of the control group was 4.67\u00a0months (95% CI 4.21\u20135.36\u00a0months); the HR of the experimental group to the control group was 0.758 . The median OS of the experimental group was 13.50\u00a0months (95% CI 12.517\u201314.949\u00a0months) and that of the control group was 12.06\u00a0months (95% CI 11.072\u201313.109\u00a0months); the HR of the experimental group to the control group was 0.817 . There were significant differences in median PFS and median OS between the experimental and control groups. The study also found that the efficacy of aflibercept was not related to previous bevacizumab treatment. This study provides crucial evidence for the approval of aflibercept combined with FOLFIRI for second-line treatment in patients with metastatic colorectal cancer who progressed or were resistant to oxaliplatin after oxaliplatin treatment.VELOUR is a proIn total, two HTA and nine SR/meta-analyses reportedA total of seven meta-analyses examined the incidence of different adverse events in patients with solid tumors treated with aflibercept. The results of six meta-analyses showed that patients with solid tumors treated with aflibercept had a higher rate of all-grade and high-grade hemorrhagic events , all-grade and high-grade proteinuria , high-grade and fatal infection , all-grade and high-grade gastrointestinal perforation , all-grade and high-grade hypertension , and the risk of treatment-related death than those in the control group which was only treated with chemotherapy. In contrast, the results of a meta-analysis showed tIn total, two pharmacoeconomic studies includedAccording to the rapid health technology assessment, for mCRC patients who were resistant or progressing after first-line oxaliplatin treatment, aflibercept combined with FOLFIRI could improve PFS, OS, and ORR compared with FOLFIRI alone. Somehow, a clinical study includedHowever, compared with FOLFIRI, aflibercept plus FOLFIRI had a higher incidence of high-grade adverse events in mCRC treatment . AdverseIn total, two cost-effectiveness analysis studies showed tvia an interactive voice response system (IVRS) using permuted-block randomization, stratified according to baseline ECOG performance status (0 vs. 1) and prior bevacizumab (yes vs. no). Between 27 July 2012 and 19 March 2014, 332 patients were enrolled and randomly assigned to treatment groups: 223 to aflibercept plus FOLFIRI and 109 to placebo plus FOLFIRI. But in this clinical study, some problems arose. Due to the clinical supply misallocation, 198 (60%) of 332 patients received at least one cycle of misallocated treatment : 122 of 223 in the aflibercept plus FOLFIRI group and 76 of 109 in the placebo plus FOLFIRI group. Finally, 111 patients received aflibercept plus FOLFIRI, 188 patients received mixed administration, and 33 patients received placebo plus FOLFIRI. The Data Monitoring Committee did not stop the study despite the misallocation. Ultimately, the researchers concluded that despite the misallocation, the study demonstrated that the addition of aflibercept to FOLFIRI chemotherapy improved PFS, overall survival, and response rate in patients from the Asia-Pacific region with oxaliplatin-pretreated mCRC. No new safety concerns were identified in this patient population. Together, these data suggest a favorable benefit\u2013risk ratio for the aflibercept plus FOLFIRI combination in this setting. This study provides some evidence for the launch of aflibercept in combination with FOLFIRI in the treatment of mCRC in China, but due to the limitations of this study, the results of this study should be treated with caution.Among the clinical studies of aflibercept combined with chemotherapy in mCRC, only one RCT was conducted in several study sites including China , which is AFLAME by At present, the guidelines for mCRC in France, the United States, and Japan recommenThis study has several advantages. It comprehensively summarizes the secondary evidence of aflibercept combined with chemotherapy for the treatment of mCRC and comprehensively analyzes the efficacy, safety, and cost-effectiveness of the therapy, which provides not only some evidence-based evidence for the clinical application of this therapy but also evidence for its drug selection decision in Hong Kong, Macao, and Taiwan regions of China.This study also has some limitations. First, it focused on the analysis of the efficacy of aflibercept combined with FOLFIRI in the second-line treatment of mCRC. Due to the lack of direct head-to-head studies, the subgroup data of the network meta-analysis included was partially adopted. The quality of the evidence may be lower than direct research. The results might be affected by publication bias. Second, this study was a rapid assessment, with mainly qualitative analysis as well as potentially limited result.Compared with FOLFIRI, aflibercept combined with FOLFIRI in the second-line mCRC treatment had better efficacy, but it was less safe and did not have a cost-effectiveness advantage. In the future, randomized controlled trials should be further carried out to clarify the effectiveness and safety of this therapy, and its effectiveness and safety should be compared with other second-line therapies such as bevacizumab, cetuximab, and regorfenib. Medical institutions in Hong Kong, Macao, and Taiwan regions of China should be cautious when using or introducing aflibercept plus FOLFIRI as mCRC treatment."} +{"text": "Dominik Stelzle was supported by the Federal Ministry of Education and Research, Germany, Grant 01KA2112B, within the Cysticercosis Network of Sub-Saharan Africa (CYSTINET-Africa). The funders had no role in the detailed study design, data collection and analysis, decision to publish, or preparation of the manuscript.The Funding statement is incomplete. It should read as follows: This study was funded by the German Research Foundation (DFG) ["} +{"text": "Branched-Chain Amino Acids (BCAAs) has been identified as a risk factor for circulatory disease. Nevertheless, the effects and mechanisms of BCAAs on the risk of moyamoya disease (MMD) remain unrecognized. Hence, we aimed to elucidate the association between circulating BCAAs and the risk of MMD and clinical subtypes.We conducted a case-control study of 360 adult MMD patients and 89 matched healthy controls consecutively recruited between September 2020 and December 2021. Serum level of BCAAs was quantified by liquid chromatography-mass spectrometry. The associations between BCAAs and risk of MMD were evaluated.P < 0.001). After adjusting for traditional confounders, the elevated BCAAs level was significantly associated with the risk of MMD . The risk of subtypes in MMD also increased with each increment in the quartiles of BCAAs. Furthermore, BCAAs offered substantial improvement in risk reclassification and discrimination for MMD and subtypes.Increased level of serum BCAAs was observed in MMD patients (Higher level of circulating BCAAs was associated with increased risk of MMD and clinical subtypes. This study will help to elucidate the pathogenesis of MMD, which may provide the support for facilitating the treatments and preventions. RNF213 variants have been identified to be associated with angiopathy in MMD, the frequency of variants was quite low in China (Moyamoya disease (MMD), characterized by progressive stenosis of distal portion of internal carotid arteries and abnormal collaterals at the base of brain, is recognized as the main cause of stroke in East Asians . MMD hasin China . Our recin China , while tRecently, progress in high-throughput multi-omics technologies has provided new insight into the pathogenesis of diseases . CirculaIn the current study, we enrolled a large population of MMD patients and healthy controls (HCs) and analyzed the characteristics of circulating BCAAs in MMD. We aimed to demonstrate the association of the serum BCAAs level with the risk of MMD and clinical subtypes. This work will help to identify novel biomarkers, and elucidate the pathogenesis of MMD, which may provide the support for improving the interventions and preventions.In this study, we prospectively recruited adult MMD patients at the Department of Neurosurgery, Beijing Tiantan Hospital from September 2020 to December 2021. Eligible patients were age 18\u201360 years, unilateral and bilateral MMD diagnosed by digital subtraction angiography (DSA) following the Japanese guidelines . Besidesvia chart views.Demographic data (age and sex), history of risk factors , clinical features , clinical manifestations were collected 1 (ApoA1), apolipoprotein B (ApoB), and homocysteine (Hcy). Hcy \u2265 15.0 \u03bcmol/L was considered as hyperhomocysteinemia (HHcy). Besides, peripheral inflammatory biomarkers including neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), platelet-to-lymphocyte ratio (PLR), systemic immune-inflammation index (SII) (PLT count \u00d7 neutrophil count/LY count), and monocyte-to-HDL cholesterol ratio (MHR) were calculated. Serum samples were also collected at baseline from all individuals. The serum samples were stored at \u221280 \u00b0C in the Central Laboratory of Beijing Tiantan Hospital. We used LC-MS techniques to quantitatively profile the serum metabolites of BCAAs. The level of BCAAs was calculated as the sum of levels of leucine, isoleucine, and valine.Fasting blood samples were collected after admission from all participants. Routine and biochemical blood tests were conducted to measure the levels of potential circulating biomarkers: white blood cell (WBC) count, lymphocyte (LY) count, neutrophil count, monocyte count, red blood cell (RBC) count, hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelet (PLT) count, glucose, creatinine, uric acid, triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein A2 test or Fisher exact test between groups, and continuous data were compared with two-tailed Student t-tests or Mann-Whitney U tests. One-way ANOVA or Kruskal-Wallis test was used to test the trend for continuous variables across BCAAs, and the Cochran-Armitage trend \u03c72 test was conducted for categorical variables. The logistic regression models were performed to identify the independent factors for MMD and its subtypes. The crude model was the unadjusted regression model of BCAAs. The model 1 adjusted for covariates including age and sex. The model 2 further adjusted for BMI, WBC count, neutrophil count, glucose, TG, TC, HDL-C, LDL-C, APO-A1, Hcy, NLR, SII, and MHR. Furthermore, we evaluated the predictive performance of models for the risk of MMD and its subtypes by establishing receiver-operating characteristic (ROC) curves and calculated the area under the curve (AUC). Moreover, the performance of BCAAs in the basic model built based on traditional risk factors were assessed. The net reclassification index (NRI) and integrated discrimination improvement (IDI) were calculated in risk classification by adding BCAAs to the basic model. P < 0.05 was considered statistical significance.All statistics analyses were performed using SPSS version 26.0 and R version 4.1.2 (R Development Core Team). Baseline characteristics were presented and compared between MMD patients and HCs. The categorical variables were presented as frequencies, and continuous variables were expressed as mean with standard deviation (SD) or median with interquartile range (IQR). Categorical data were compared using the \u03c7A total of 360 MMD patients and 89 matched HCs were included in the study.P < 0.05 for all). In MMD patients, the levels of systolic blood pressure (SBP), diastolic blood pressure (DBP), and BMI were significantly higher than in HCs. Patients in groups of MMD subtypes had a higher level of WBC count, neutrophil count, glucose, TG, Hcy, NLR, SII, and MHR than in HC group . Levels of laboratory results including LY count, HGB, HCT, MCHC, TC, HDL-C, LDL-C, ApoA1, and PLR were significantly different between groups . In addition, patients with MMD and its subtypes had a significantly higher level of BCAAs than that of HCs , while patients with hemorrhagic-type MMD had a lower level of BCAAs than that of infarction-type (P < 0.05) (Baseline characteristics of MMD cases and HCs were shown in < 0.05) . The sig < 0.05) .Clinical characteristics of MMD patients and HCs according to the BCAAs quartiles were shown in 1, Hcy, NLR, SII, and MHR, cases in Q4 of BCAAs were significantly associated with a higher risk of MMD than those in Q1 . The ROC curves with AUC of models for the occurrence of MMD were constructed in P = 0.025; OR 3.95, 95% CI 1.20\u201313.06, P = 0.024, respectively). Q4 of BCAAs was significantly associated with the risk of infarction-type and hemorrhagic-type MMD compared with Q1 in Model 2 . In contrast to the Crude model and Model 1, the Model 2 consistently showed prominent improvements in the predictive values of subtypes of TIA, infarction, and hemorrhagic MMD .Besides, the risk of MMD and its subtypes increased with each increment in the quartiles of individual BCAA . SimilarWe compared the performance of different models for predicting the risk of MMD and its subtypes . The addIn this large case-control study of 449 participants, we firstly investigated the association of serum metabolite of BCAAs with the risk of MMD and clinical subtypes. We identified that the serum level of BCAAs was significantly higher in MMD patients than in HCs. The elevated level of BCAAs was strongly associated with increased risks of MMD and subtypes. Collectively, our findings outlined the crucial relevance of increasing serum BCAAs with the risk of MMD.Amino acids are important nutrients for humans. As the precursors of proteins, amino acids participate in various life activities and metabolism . BCAAs iAs nutrient signaling molecules, BCAAs mainly transduce the mTOR pathway . BCAAs, BCAAs may generate a chronic inflammatory response by increasing the expression of pro-inflammatory cytokines , leading to changes in endothelial and smooth muscle cell phenotypes, and thereby producing the pathological conditions. Some studies have identified that isoleucine was positively correlated with IL-6, endotoxin and oxLDL, while leOur study showed that several factors were associated with the quartiles of BCAAs. There were growing trends of diabetes and BMI along with the level of BCAAs. It has been verified that BCAAs was related to the metabolic disorders, including diabetes and obesity . AlthougIn this study, total and individual BCAAs were all analyzed. The differences among three individual BCAAs tended to be similar. Although BCAAs is a compound of three substances, each BCAA may have distinct effects. Isoleucine and valine, while not leucine, mediated the metabolic health . It is oin vitro or in vivo experiments, and larger prospective cohort studies with follow-up outcomes are warranted to reveal the effect and mechanism of BCAAs on the pathogenesis of MMD.Several limitations should be considered in this study. First, this is a single-center study with relatively small sample size. Although the potential bias was inevitable, this is the largest study to investigate the association of serum BCAAs and MMD. Second, the study was conducted in a Chinese population with adult MMD, the findings may not be generalized to the overall populations of MMD. Third, the information of patient-level diets was not included in the study. The pattern of diets may have an impact on the outcomes. Fourth, the results of serum metabolites are affected by many factors. Although the confounders have been adjusted in the regression models, we can only demonstrate the association of serum BCAAs with the risk of MMD. Further Our study indicated that higher circulating BCAAs level was associated with increased risk of MMD and clinical subtypes. This work will help to elucidate the pathogenesis of MMD, which may provide the support for facilitating the interventions and preventions.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Beijing Tiantan Hospital, Capital Medical University. The patients/participants provided their written informed consent to participate in this study.Study concept and design: CZ and PG. Collection and assembly of data: CZ, PG, CL, XYu, and YZhai. Data analysis and interpretation: WL, QH, JL, XL, and JW. Manuscript writing: CZ. Manuscript revision: XYe, QZ, RW, and YZhang. Provision of study materials: JZ and DZ. Final approval of manuscript: all authors.This study was supported by National Key Research and Development Program of China (2021YFC2500502), National Natural Science Foundation of China (81701137 and 81870904), Beijing Municipal Commission of Education (KM201910025014), and Beijing Municipal Administration of Hospitals' Mission Plan (SML20150501).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Psychotic experiences (PE) occur most often in childhood, at the same age many mental disorders (MD) develop. There is growing evidence that those who report PE and MD show poorer health outcomes. If this occurs in psychosocial outcomes e.g. self-esteem, stress, mental distress, or social support, is under examined. Attachment anxiety and avoidance are the dimensions of attachment, which is hypothesized to develop in infancy as a mechanism for interpersonal relationships in times of need.To examine the role of transient childhood PE in adult psychosocial outcomes, in those with and without MD. Additionally, to examine if the dimensions of attachment attenuate this model.One hundred and three participants attended baseline (age 11 \u2013 13) and 10-year follow-up. PE and MD were collected using the Schedule for Affective Disorders and Schizophrenia for School-aged Children, Present & Lifetime Version. Attachment and outcomes were collected using self-report measures. Analysis compared those with PE, MD and PE and MD, to healthy controls.PE in childhood was associated with lower self-esteem and lower perceived social support from friends. Lower self-esteem in adulthood was more pronounced in those reporting PE and MD, and was additionally associated with stress in relationships, daily life, and mental distress. Childhood MD without PE was not significantly associated with any psychosocial outcomes. Attachment dimensions significantly attenuated the relationship between PE and self-esteem.This paper illustrates the significant association of childhood PE on adult outcomes, independent of the effect of co-occurring MD, and demonstrate attachment dimensions role in this model.No significant relationships."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Mobile learning is one of the innovative teaching techniques that help medical students gain knowledge and skills. One of the factors that expanded theuse of this strategy was the COVID-19 pandemic. However, the educational pedagogy of such technology has been neglected. This article aimed tocritically review available mobile learning models in medical education to suggest a comprehensive model in the field of mobile learning. We conducted this critical review based on the five steps of the Carnwell and Daly method. For a comprehensive systematic search from 2000 to April 2021,the following keywords were used: Personal Digital Assistant, m learning, Mobile learning, Ubiquitous learning, U learning, medical students,and medical education. 3176 studies in PubMed, Scopus, ERIC, Magiran, and Web of Science were identified. In total, 8 articles entered the study. Eight models of mobile learning in medical education were identified. The key features of each model were extracted and integrated into thenew model for the successful design and implementation of mobile learning. This model includes three main elements of mobile learning: 1-stakeholders,2-interaction, and 3-technology, which are influenced by external factors including Mobiquette, legitimacy, and awareness. The results of this study are an important contribution to the knowledge collection in mobile learning in medical education.We introduced a comprehensive model of mobile learning including specific characteristics of strategies in the context of medical education. Recently, higher education has undergone extensive changes due to technological advances . FollowThere are many definitions of m-Learning. In 2003, Brown defined m-Learning as an extension of e-learning . HardenThe learners use mobile for their daily activities, so they are tend to apply them in their educations . This aSome evidence has indicated that m-Learning has the potential to improve the use of evidence-based decision-making . In theSeveral prominent medical universities such as Oxford , HarvarThe COVID-19 pandemic caused an expansion in the use of m-Learning , 25. DSeveral studies have been conducted to evaluate the practical application of mobile devices and computers in medical education;however, literature is scarce on the educational pedagogy needed to use this technology , 30. IThere are several outstanding m-Learning models in higher education , but inThis study applied the method of critical review based on the \u2018Five-phase method\u2019 adopted by Carnwell and Daly. The five steps are: a) determining the scope of review, b) recognizing relevant information sources, c) reviewing the evidence, d) applyinga general and critical perspective in writing, and e) concluding the literature for further studies . The reAccording to Hart , a critWhat research has been done on m-Learning models in medical education and what are the gaps?Which key elements should be considered for m-Learning model in the medical education system?This review included reports and peer-reviewed articles related to m-Learning from 2000 to April 2021 that were retrieved in the databases PubMed, Scopus, ERIC, Magiran, and Web of Science.The ontological search was keywords related to m-Learning: Personal Digital Assistant, m learning, Mobile learning, Ubiquitous learning, U learning, medical students, and medical education.Titles and abstracts were screened by two independent researchers to determine the relevance. The full-text versions of theincluded materials were reviewed. In case of doubts regarding eligibility, a third researcher was consulted to resolve any disagreements .Gray literature identifies by hand-searching through conference proceedings, theses, and abstracts.The search identified 3176 articles. 2739 articles were removed since the title, keywords, or abstract did not demonstrate thedesired concepts. At the eligibility step, 68 full texts of documents remained, and then 60 articles were excluded based on theinclusion/exclusion criteria. Finally, 8 articles were included .Eight models of m-Learning in medical education were identified. The extracted data included authors, year of publication,country, model components, participants, sample, and outcomes .This study was approved by the Research Ethics Committees of School of Medical Education affiliated to Shahid Behshti Universityof Medical Sciences, Tehran, Iran with the code of IR.SBMU.SME.REC.1400.026.The final review included eight models, primarily rooted in developed countries. Davies et al. developBriz-Ponce et al. consideJoynes et al. also de\"Maturity of learning\" is related to how senior students demonstrated greater maturity in using resources than junior students.Another component, \"personalization\" is about students adapting the available resources to tailor their own needs.The concept of \u2018learning legitimately\u2019 is key to success of m-Learning. Participants indicated that legitimacy, as the mandatory nature of theprogram, has been a factor in encouraging them to use m-Learning.\"Developing professional identity\", based on the participant's experience, is the use of mobile resources at the undergraduatelevel leaving a lasting impact after graduation and causing maturity in their behavioral patterns of learning in their work-life as health professionals.The component \"learning differently\" was at the core of the model. Personalization, learning legitimately, maturity of learning,and developing professional identity revolved around the core component. M-Learning permits students and faculty to gain various learning experiences.One of these experiences improved their ability to \"personalize\" mobile resources for learning. Another experience was that theparticipants learned how to maturely use the mobile resource in the workplace over time.Koohestani et al. proposeMotivational factors (negative and positive) cause negative and positive attitudes in students, which affect the behavioral intentionand situational reaction. Students understand the benefits of using m-Learning in the learning path, which causes them to reflect, and ultimately this reflection affects motivational factors.Aliano designePerceived gratification means that working in an environment with mobile technologies results in improved motivation causing greater personal satisfaction.Thus, the process of learning becomes more enjoyable, provoking greater interest in the students. Some of these components are identical tothe components of the model by Briz-Ponce , exceptOne of the salient features of this model is that it examines the relationship between socio-demographic data and other components.Age and perceived usefulness had an inverse relation, similar to the perceived ease of use and the perceived gratification variable. The modified model FRAME for medical and nursing education context was introduced by Lall et al. . The FRIn the modified model FRAME, social technology was changed to make up for the impact of mobile technologies on social interaction. They also added a circle that covers three circles named implementation.Insufficient institutional resources, lack of training and support, and limited planning and management of m-Learning seem to be thekey to understanding how m-Learning for medical students might be implemented.Kucuk proposeIn this model, perceived usefulness, which means personal beliefs that individuals obtain success in their performance when they usepertinent technology, affects the attitude to use m-Learning. Also, subjective norms, beliefs about whether most people agree or disagreewith the behavior, are influenced by students\u2019 readiness. Perceived behavioral control is mainly influenced by perceived self-efficacy.This means that if students make sure they use mobile apps, they will have behavioral control and intend to use it for m-Learning purposes. Mosalanejad suggestEight m-Learning models were extracted from 8 reviewed articles and the key elements of each model were described. In this section, based on the fourth stepin the method of Carnwell and Daly, the critics' views (if any) were reviewed, and at the end, our views are explained .Davies and colleagues answered questions such as how medical students use mobile technologies. One of the strengths of thisstudy is that m-Learning is used in formal medical education and data were collected in both quantitativeand qualitative ways . AnotheThe second model introduced by Briz-Ponce and colleagues attempts to provide insight into the factors that may affect theacceptance of mobile devices and applications by students and medical professionals in medical education . Few stNiazazari and colleagues acknowledged that in addition to the above points, the location and type of mobile device affected the acceptance of m-Learning . AL-EmrIn 2016, Joynes and colleagues developed a model for m-Learning by examining the views of students and educators on the impact of MBChB Mobile . In thiAnother strength of this model is that it is not considered as an alternative to conventional methods. Instead, mobile resourcesalign and complement the curriculum by adding different learning options. Thus, m-Learning integrates into the curriculum and is also a kind of blendedlearning creation. Endorsing this model, Green and colleagues believe that blended learning creates a rich and engaging experience for students .In the mentioned model, only students and teachers were considered stakeholders. Although this model is based on practical faculty experiences, the features of content, educational design, and comprehensive evaluation methods used are not mentioned in the model. Also, in the implementation of this model, factors such as network connection and ease of access to content, technical support team, and costs have been considered, but in the m-Learning model, they were not.Koohestani and colleagues in 2019 used a qualitative method to design an m-Learning model for medical students according to local conditions and contexts . This mAccording to this model based on the informal experiences of students and teachers of m- learning, the operational aspects of m-Learning implementation, including the design of educational content, content features, and students' assessment and the use of m-Learning along with traditional education have not been considered. The results of this study have been published, and so far it has not been referenced or criticized.In the fifth model, Alia\u00f1o et al. showed the factors influencing the acceptance and intention of using mobile devices as learning resources in medical education . The suIn this model, the individual characteristics and demographics of the student are mentioned in detail; students are considered the only stakeholders. Other elements such as learning context, interaction with other stakeholders, use of m-Learning along with traditional education, educational design, mobile content features, as well as student assessment are not mentioned in the design and implementation of m-Learning. In terms of technology, network connection, technical support, and the cost of equipment and facilities are not mentioned.Lall et al. introduced a modified FRAME model by examining the factors that facilitate or hinder the implementation of m-Learning strategies in medical and nursing education . KhosraIn the seventh model, Kucuk and colleagues proposeHowever, factors such as the performance of various m-Learning activities independent of the medical course by students may limit the use of this model to design the environment and implement effective m-Learning in medical education. Another point is that in this model, factors such as educational content design, content features, and student assessment, and the use of m- learning along with traditional education, the context of learning are not considered. Also, the technology aspect is generally neglected in this model.Mosalanejad and colleagues presented a model based on the factors affecting m-Learning . One ofAnother feature of this model is that the needs of students, as one of the main stakeholders, are included.No mentioning of other stakeholders, use of m-Learning along with traditional education, factors such as educational design,mobile content, student assessment, technical support, ethical concerns about patient privacy, and data security are also overlooked in this model.The eight models discussed above point to some of the features of m-Learning, such as usability, collaboration, and flexibility,while ignoring other important features. The key features of each model were extracted and integrated into the new model for a successful designand implementation of m-Learning. This new model is called K-ASK3. The K-ASK3 model contains all the elements of the other models and hasthe necessary comprehensiveness. This model includes three main elements of m-Learning: 1. Stakeholders, 2. Interaction, and 3. Technology,influenced by external factors including mobiquette, legitimacy, and awareness of m-Learning. The three main elements refer to the principles of m-Learning pedagogy. The first element is stakeholders. Students, teachers, peers, education administrators, educational designers, and technical experts,family or caregivers, patients, and other health professionals are stakeholders in m-Learning and have a significant impact on itssuccessful design and implementation. These people communicate and collaborate using the flexibility offered by mobile technologies - 57 .The second element is interaction. It includes the educational aspect of m-Learning and interactions between people, devices, and systems.Also, it refers to the characteristics of m-Learning that help the students and teachers to interact with each other in terms of cooperation,blended learning, educational design of m-Learning in the field , 58 .The third element is technology. Technology provides access to learning resources anywhere and anytime.In learning environments, technology plays a mediating role in improving learning comprehension. This element shows the featuresassociated with mobile devices, including network connectivity, flexibility, usability, technical support, reliability,and costs associated with the technology , 57 .Other important components of the model introduced above are external factors.1-Mobiquette: To maintain the confidentiality of patient information, teaching behavioral etiquette of using mobile devicesin the clinic to medical students is a necessity , 33 .2-Legitimacy: The medical education institute supports the use of mobile resources in different places .3-Awareness of m-Learning: Awareness of students and faculties of m-Learning and its benefits affect their intention to use this type of education , 41Fi. One of the limitations of this study is the limited number of studies in medical sciences and the lack of a similar structure or model toguide the study. Due to the use of only English and Persian articles, some valid documents may not be included in this study.The main focus of the study was on innovative conceptual interpretation of researchers. Another limitation is the low sensitivity of the searches.For this limitation, the authors used experienced researchers. On the other hand, our study has some strengths; we mentioned theIranian model of m-Learning in our study and used the researchers\u2019 expertise and experience in assessing the quality to increase scientific rigor.Future research can explore the impact of m-Learning on the acquisition of knowledge, attitudes, and skills of medical students in theCOVID-19 and post-COVID eras, and its role in theoretical and clinical education.The results of this study will be an important contribution to the knowledge collection in the field of m-Learning in medical education.Reviewing the models shows that each model tries to explain a part of the m-Learning strategies that have not been represented in other models.Therefore, we introduce a comprehensive model of m-Learning (K-ASK3 model) that includes specific characteristics of strategies in themedical education context. The K-ASK3 model contains all the elements of the other models. This model includes three main elementsof m-Learning: 1. Stakeholders, 2. Interaction, and 3. Technology, influenced by external factors including mobiquette,legitimacy, and awareness of m-Learning. Paying attention to the elements of this model to change the educational policies of institutionsmay play an important role. Faculty development for using m-Learning should be included in the work program of educational institutions.The authors would like to thank their colleagues, who provided them with supplemental data.M.K, M.M.S, S.A, P.K, M.K, N.Kh contributed to the conception and design of the work; the acquisition, analysis, or interpretation of data for the work. All Authors contributed indrafting and revising the manuscript critically for important intellectual content. All authors have read and approved the final manuscript and agreeto be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.None declared."} +{"text": "Chronic subdural hematoma (CSDH) is common in elderly people with a clear or occult traumatic brain injury history. Surgery is a traditional method to remove the hematomas, but it carries a significant risk of recurrence and poor outcomes. Non-surgical treatment has been recently considered effective and safe for some patients with CSDH. However, it is a challenge to speculate which part of patients could obtain benefits from non-surgical treatment.To establish and validate a new prediction model of self-absorption probability with chronic subdural hematoma.The prediction model was established based on the data from a randomized clinical trial, which enrolled 196 patients with CSDH from February 2014 to November 2015. The following subjects were extracted: demographic characteristics, medical history, hematoma characters in imaging at admission, and clinical assessments. The outcome was self-absorption at the 8th week after admission. A least absolute shrinkage and selection operator (LASSO) regression model was implemented for data dimensionality reduction and feature selection. Multivariable logistic regression was adopted to establish the model, while the experimental results were presented by nomogram. Discrimination, calibration, and clinical usefulness were used to evaluate the performance of the nomogram. A total of 60 consecutive patients were involved in the external validation, which enrolled in a proof-of-concept clinical trial from July 2014 to December 2018.CI, 0.637\u20130.788)] and good calibration (p = 0.986). The nomogram in the validation cohort still had good discrimination and good calibration (p = 0.441). A decision curve analysis proved that the nomogram was clinically effective.Diabetes mellitus history, hematoma volume at admission, presence of basal ganglia suppression, presence of septate hematoma, and usage of atorvastatin were the strongest predictors of self-absorption. The model had good discrimination [area under the curve (AUC), 0.713 (95% This prediction model can be used to obtain self-absorption probability in patients with CSDH, assisting in guiding the choice of therapy, whether they undergo non-surgical treatment or surgery. Chronic subdural hematoma (CSDH) is common in elderly people, which is combined with clear or occult traumatic brain injury (TBI) history . Its incAll patients had provided written consents before enrollment. This study re-analyzed the entire database with the approval of the ethics committee of Tianjin Medical University General Hospital. The primary cohort of this study from a randomized controlled trial (ATOCH) , which wWe included patients with the following characteristics: no surgery treatment, first onset of CSDH, and lack of anticoagulant treatment. The following exclusion criteria was adopted to all study samples: CSDH caused by cancer, hemopathy, or other known serious comorbidities, and lack of follow-up at the 8th week.The following clinical statistics were gathered: age, sex, history of TBI, history of hypertension, diabetes mellitus and hypeCT images were collected with different scanners at each center and sent electronically to Tianjin Medical University General Hospital. All the images were uniformly analyzed by 3 independent neuroradiologists to standardize the measurements. The following imaging data were record: hematoma volume , thickness of hematoma, midline shift distance, presence of basal ganglia compress, hematoma location , and presence of septate hematoma . HematomWhether hematoma could be self-absorbed without surgery at the 8th week was the outcome of this prediction model. Good absorption was defined as hematoma reduction >50% from baseline hematoma volume. Poor absorption was defined as hematoma reduction <50% or hematoma enlargement.t-test, Wilcoxon test, or chi-square test. All statistical analyses were conducted using the SPSS version , with p < 0.05 given statistical meaning.Following the evaluation of the normal distribution by Shapiro\u2013Wilk testing, the data were summarized using the mean with SD or the median with quartiles as continuous variables, and the classification variables were summarized using the frequency and percentage. The comparison between good absorption and poor absorption groups in the primary and validation cohorts was analyzed by Student's unpaired The least absolute shrinkage and selection operator (LASSO) method was usedCalibration curves were drawn to evaluate the alignment of the nomogram by Hosmer\u2013Lemeshow tests . A receiThe validation cohort helped to inspect the performance of the internally validated nomogram. The predictive model formed in the primary cohort lends itself to all patients in the validation cohort and helps to digitalize the total score for every single patient. The ROC curve and calibration curve were then performed on the factors of the total score.A decision curve analysis was managed to evaluate the clinical effectiveness of the nomogram, which quantified the net benefits at different threshold probabilities in the primary cohort .In Known predictors, such as baseline volume and drug usage were associated with greater likelihood of good absorption. Of these features, 15 potential predictors of 196 patients in the primary cohort is about 65, the point of DM is 0, the point of separate is 0, and the point of drug is about 8. The total point is 73, indicating that self-absorption is approximately 60%.p = 0.986), which indicated that the perfect fit did not deviate. The ROC curve of the prediction nomogram indicated that the AUC was 0.713 in the primary cohort.The calibration curve of the nomogram with good absorption probability showed good consistency between prediction and observation in the primary cohort . The Hosp = 0.441). The ROC curve for the prediction of good absorption showed that the AUC was 0.709 in the validation cohort.A good calibration of good absorption probability was observed in the validation cohort . No signThe decision curve for the nomogram showed tA nomogram was established and validated for the personalized prediction of hematoma self-absorption in patients with CSDH. The primary cohort was a well-designed multicenter CSDH clinical trial population, which could present the patients with CSDH in China. The nomogram incorporated five items: history of diabetes mellitus , the useFor examining the predictor-outcome association, the 15 candidate features were eventually reduced to 5 potential predictors by using the LASSO method to narrow the regression coefficients. This method went beyond the method of selecting predictors based on the univariate intensity associated with the outcome . The nomAlthough the pathogenesis of CSDH was studied by a succession of studies, which aspects included pontine vein avulsion hemorrhage, increased osmolality, hematoma capsule bleeding, and local hyperfibrinolysis, the pathogenesis and absorption mechanism of CSDH were not well understood , 15. TheIt is well-known that diabetes mellitus is a prominent risk factor for coronary heart disease and stroke . Our resStatins, also known as selective HMG-CoA reductase inhibitors, have previously been applied to treat hyperlipidemia. They have been widely used in the treatment of cardiovascular disease. Currently, some studies had proved that atorvastatin could promote the absorption of CSDH and prevent the recurrence of CSDH \u201327. Our The hematoma capsule is very important in which both bleeding and reabsorption occur. El-Kadi et al. demonstrThe most important part of using the nomogram is the requirement to explain operations clearly based on individual need. The difficulty we face lies in being unable to capture the clinical consequences of a particular level of discrimination or degree of miscalculation , 31. TheSome limitations should be considered in this paper. First, to maximize the enrollment of clinical trials , which as the primary cohort, we selected CSDH patients with relatively small baseline hematoma volume. Hematoma volume is known as the main influenced factor of hematoma absorption , which wIn conclusion, this study proposed a nomogram that combined clinical features and CT image features, which could be conveniently used to predict the probability of CSDH self-absorption and reduce unnecessary manipulation.Tianjin Medical University General Hospital: Shuyuan Yue. First Affiliated Hospital of Harbin Medical University: Shiguang Zhao. Peking Union Medical College Hospital: Renzhi Wang and Junji Wei. Southwest Hospital: Hua Feng and Rong Hu. Second Affiliated Hospital, Zhejiang University School of Medicine: Jianmin Zhang. Qilu Hospital of Shandong University: Xingang Li. Huashan Hospital Fudan University: Ying Mao and Jin Hu. Xiangya Hospital of Central South University: Xianrui Yuan. Xijing Hospital of the fourth Military Medical University: Zhou Fei. Beijing TianTan Hospital, the Capital Medical University: Yuanli Zhao. General Hospital of Chinese People's Liberation Army: Xingang Yu. Prince of Wales Hospital, Chinese University of Hong Kong: Wai Sang Poon. Linyi People's Hospital: Xinde Zhu. Jiangsu Provincial Hospital, Nanjing Medical University First Affiliated Hospital: Ning Liu. First Affiliated Hospital of Fujian Medical University: Dezhi Kang. General Hospital of Ningxia Medical University: Tao Sun. Second Affiliated Hospital of Hebei Medical University: Baohua Jiao. First Affiliated Hospital of Zhengzhou University: Xianzhi Liu. Affiliated Hospital of Xuzhou Medical College: Rutong Yu. Central Hospital of Erdos: Junyi Zhang. Xi'an Tangdu Hospital of the fourth Military Medical University: Guodong Gao. First Affiliated Hospital of Shanxi Medical University: Jiehe Hao. Provincial People's Hospital of Inner Mongolia: Ning Su. Cangzhou Central Hospital: Gangfeng Yin. Second Affiliated Hospital of Nanchang University: Xingen Zhu. Shanghai Changzheng Hospital: Yicheng Lu. 117th Hospital of Chinese People's Liberation Army: Jianrong Li.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Tianjin Medical University General Hospital. The patients/participants provided their written informed consent to participate in this study.JS, RJ, and JZ had full access to all of the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis. RJ and JZ: concept and design. YT, DW, and XZ: acquisition, analysis, or interpretation of data. YT: drafting of the manuscript. HW, YW, and SA: critical revision of the manuscript for important intellectual content. CG and JH: statistical analysis. YT, DW, RJ, and JZ: obtained funding. All authors contributed to the article and approved the submitted version.http://www.zycmsf.com/en/index.asp).The National Natural Science Foundation of China, 81971173 and 81501057, YT received the funding. The National Natural Science Foundation of China, 82171359 and 81671380 and Beijing-Tianjin-Hebei Cooperation Project, 19JCZDJC64600(Z), DW received the funding. The State key Program of the National Natural Science Foundation of China, 81930031, JZ received the funding. Tianjin Research Program of Application Foundation and Advanced Technology, 19YFZCSY00650, RJ received the funding. ATOCH trial was supported by the Chinese Society of Neurosurgery, Chinese Medical Association and Zhao Yi-Cheng Medical Science Foundation (The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "To the Editor,Telemedicine is a term that refers to any activity of medical nature that involves communication despite being physically distinct . In otheInflammatory bowel disease (IBD) is characterized by gastrointestinal illnesses that are chronic or remitting inflammatory diseases. IBD is further subdivided into two subtypes: ulcerative colitis (UC) and Crohn's disease (CD) . Recent There are numerous difficulties in the timely diagnosis, treatment adherence, financial burden, regular follow ups, awareness, and misdiagnosis of inflammatory bowel disease . All theAll the major issues that are hindering the control of IBD can be addressed with telemedicine. Telemedicine interventions have shown a positive role in improving quality of life and reducing the number of required clinical visits in patients with IBD . TelemedIn short, telemedicine is the most cost-effective, time-saving, and patient-friendly method to promoting patient compliance. Telemedicine can adequately resolve all problems that are encountered in the diagnosis and management of IBD patients. Investment into the necessary infrastructure to make telemedicine more accessible is essential at the government level. A reduction in the cost of telemedicine devices will be conducive to the widespread adoption of telemedicine in clinical practice. Healthcare workers and patients need to be trained to operate telemedicine devices optimally. Collaboration between all stakeholders is critical to ensure a smooth transition to telemedicine-based management of IBD patients.Not applicable.None.Faizan Fazal: Study conception, write-up, critical review and approval of the final version.Mohammad Ebad Ur Rehman: Study conception, write-up, critical review and approval of the final version.Omaima asif: Study conception, write-up, critical review and approval of the final version.Haris Mustafa: Study conception, write-up, critical review and approval of the final version.Usama Tanveer: Study conception, write-up, critical review and approval of the final version.Tehseen Haider: Study conception, critical review and approval of the final version.Abdul Rauf Khalid: Study conception, write-up, critical review and approval of the final version.Sajeel saeed: Study conception, write-up, critical review and approval of the final version.Jawad basit: Study conception, critical review and approval of the final version.1.Name of the registry: Not applicable2.Unique Identifying number or registration ID: Not applicable3.Hyperlink to your specific registration (must be publicly accessible and will be checked): Not applicableFaizan Fazal.Mohammad Ebad Ur Rehman.Not applicable.Not commissioned, externally peer reviewed.The authors have no conflicts of interest."} +{"text": "With the advances of the Belt and Road Initiative (BRI), the geo-economic interactions between China and countries along the 21st-Century Maritime Silk Road counties (MSRCs) continue to increase, and analyzing the geo-economic relations between China and the MSRCs is extremely important for a scientific understanding of bilateral geo-economic cooperation. Differently from the traditional logic of geo-economic competition and cooperation, we constructed a new framework based on the dominant factors of geo-economic relations and used an extreme random forest regression model to classify the geo-economic relation types between China and MSRCs from 2006 to 2017. The results show that the unbalanced development of investment and trade between China and MSRCs hindered the enhancement of the intensity of bilateral geo-economic linkage from 2006 to 2017. The \u201cMatthew effect\u201d of China\u2019s geo-economic flow linkage with MSRCs is significant. There are obvious differences in the dominant factors affecting the types of geo-economic relations between China and MSRCs, and the distribution of the importance of the indices of the types of geo-economic relations in each country is disordered. Geopolitics, markets, and resources have played important roles in the geo-economic linkages between China and MSRCs. There are five types of geo-economic relations between China and the MSRCs: market-oriented type, resource-oriented type, market-resource-oriented type, market-geopolitics-oriented type, and resource-geopolitics-oriented type, of which the market-oriented type is the most important type of geo-economic relations. In the future, China should focus on regional powers along the Maritime Silk Road for bilateral geo-economic cooperation, actively promote the balanced development of bilateral geo-economic elements flows, strengthen geopolitical cooperation with MSRCs, and formulate cooperation plans according to the types of geo-economic relations. In the geo-economic era of globalization, the interactions and transformation of geo-economic factors are more rapid, geo-economic networks have become larger and more complex, and geo-economic interactions between countries are more complex than ever before . As the Through the 21st-Century Maritime Silk Road, China is engaged in broader, higher-level, and deeper regional cooperation, and is working to build a mutually open, inclusive, balanced, and beneficial geo-economic cooperation architecture . In the To scientifically reveal the changes in the geo-economic linkages and their types between China and MSRCs, we constructed a new analytical framework of geo-economic relations. Firstly, this study takes MSRCs as the research object and constructs the geo-economic flow linkage intensity model based on flow space theory to analyze the evolution of the characteristics of geo-economic linkages between China and MSRCs. Then, an index system of geo-economic relation types is constructed, and the extreme random forest regression model is used to identify the importance of the geo-economic relation type indices and determine the dominant factors of geo-economic relations between China and MSRCs. Finally, this research divides the geo-economic relations between China and MSRCs into different types based on the importance of the geo-economic relation type indices.The design of this study is as follows. In Since its birth, the concept of geo-economy has had a strong geopolitical color. When Luttwak proposed the concept of the geo-economy, he emphasized that economic power and economic means are important aspects of the game between powerful countries and the distribution of power . BlackwiGeo-economic competition and cooperation, and geo-economic relations matching are the key topics of geo-economic linkage research. Geo-economic competition and cooperation mainly focus on the competitiveness, cooperation, and complementarity of geo-economic relations. The geo-economic flow between China and the countries along the Belt and Road is increasing, and the geo-economic relations between China and these countries are continuing to improve . After CGeo-economic elements play an important role in the changing of geo-economic linkages. Geo-economic factors such as resources, trade, finance, and technology are shaping the competition and cooperation of today\u2019s geo-economy , as resoThe ways in which various influencing factors act on geo-economic linkages extend from resource and political tools to technical tools . DiffereIn the process of constructing the BRI, the flow of geo-economic elements such as trade, transportation, and resources is being drawn into a new map of geo-economic linkages among the MSRCs . In termThere is no doubt that the perspectives and content of geo-economic research are becoming increasingly diversified, and geo-economic elements such as resources, trade, finance, and technology play key roles in shaping geo-economic linkages. The construction of the Belt and Road has created closer and more frequent geo-economic exchanges between China and MSRCs than before, and bilateral geo-economic linkages are also showing new characteristics. However, the existing studies are limited to the logic of geo-economic competition and cooperation when analyzing the geo-economic relations between China and MSRCs, and have not clarified the dominant factors affecting the evolution of geo-economic linkage between China and MSRCs, nor do they propose a method to classify the types of geo-economic relations between China and MSRCs. Therefore, this study addresses the following three questions: (1) How do the geo-economic linkages between China and MSRCs evolve? (2) What factors drive the evolution of geo-economic relations between China and MSRCs? (3) How can one classify the types of geo-economic relations between China and MSRCs? To solve these problems, we constructed a new analytical framework to analyze the characteristics of the geo-economic linkages and geo-economic relation types between China and MSRCs based on geo-economic element flow.The flow space theory of Castells pointed out that the dominant form of space is no longer the local space in the network society, but a new flow space . In the ijL is the geo-economic flow linkage intensity between i and j, iT is the trade flow from i to j, jT is the trade flow from j to i, iI is the investment flow from i to j, jI is the investment flow from j to i, rp is the political geopolitical variable, rd is the transportation geopolitical variable, and iL is the total geo-economic flow linkage intensity in i.In related studies, geo-economic linkage intensity is often measured using gravity models and gray correlation analysis . HoweverijL, the closer the geo-economic relations. The geo-economic flow linkage intensity model objectively evaluates the strengths and weakness of geo-economic interactions among countries based on geo-economic element flow. It covers the most important investment flows and trade flows in geo-economic linkages. It can not only reveal the vector characteristics of geo-economic flows between countries, but can also reveal the evolution characteristics of geo-economic relations.The geo-economic flow linkage intensity reflects the closeness of geo-economic relations among countries. The greater the value of 2), mean square error (MSE), mean absolute error (MAE), and explained variance score (EVS) are used to test the effect of random forest regression estimation.The random forest is an integrated algorithm based on the decision tree proposed by Breiman , which iThe MSRCs have no precise spatial scope as part of the BRI . HistoriThe intensity of the geo-economic linkages between China and MSRCs was measured by the geo-economic flow linkage intensity model, and it was found that the intensity of geo-economic linkages between China and the MSRCs rose from 282,836.59 million dollars in 2006 to 833,803.57 million dollars in 2017 . This shThe geo-economic relations between China and the MSRCs have been greatly impacted by the financial crisis, global economic adjustments, and recovery of trade protectionism. Due to these factors, the geo-economic flow between China and the MSRCs fell in 2008, 2012, and 2016; the geo-economic competition intensified; and the geo-economic linkage intensity decreased. Since the construction of the BRI, the intensity of China\u2019s geo-economic linkage with the MSRCs has not increased rapidly, but it has increased in volatility. On the one hand, under the influence of the global trade situation and economic growth, China\u2019s trade and investment relations with the MSRCs have increased in volatility. On the other hand, the development of investment and trade between China and the MSRCs is unbalanced. China\u2019s investment in the MSRCs is far greater than China\u2019s investments in China, and China\u2019s exports to the MSRCs are far greater than their exports to China. This imbalance hinders the improvement of the intensity of the geo-economic linkages between China and the MSRCs to a certain extent.In order to reveal the evolution of geo-economic linkage intensity between China and MSRCs, we selected 2006, 2010, 2014, and 2017 as time nodes and used ArcGIS software to visualize them . AccordiCountries with high geo-economic linkages with China are concentrated in Southeast Asia, Japan, and South Korea. These countries are surrounded by China, Japan, and South Korea, which have large economic volumes, broad ASEAN markets, strong geo-economic complementarity, and frequent bilateral trade and investment exchanges. In addition, bilateral geo-economic growth has been sustained by the construction of the China\u2013ASEAN free trade area and the China\u2013South Korea free trade area. Over time, the pattern of geo-economic relations between China and the five Balkan Peninsula countries, four African countries, and fourteen West Asian countries has been relatively stable; and the pattern of geo-economic relations between China, Southeast Asia, and South Asia has changed greatly. From 2006 to 2010, the geo-economic connection intensity between India and China rose from a low to a high level. Since then, the growth rate of the strength of Sino-Indian geo-economic ties has been slow, and the strength of bilateral ties has declined, being affected by factors such as different development stages, trade imbalances, and geopolitics. According to the intensity ranking of geo-economic linkages, the intensity of geo-economic linkages between China and Vietnam increased the fastest, and that between China and Syria declined the fastest. From 2006 to 2017, Vietnam rose from 12th to 3rd, and Syria dropped from 28th to 39th. The industrial structure and resource endowments of China and Vietnam are highly complementary; a large number of Chinese enterprises invest in factories in neighboring Vietnam. Bilateral trade flows and investment flows have increased rapidly, from 2748.04 million dollars in 2006 to 64,837.86 million dollars in 2017, the largest increase in all countries. The investment and trade between China and Syria were seriously damaged by the Syrian War, which has continued since 2011. From 2003 to 2017, Syria\u2019s exports to China dropped from 506,637.16 million dollars to 13,341.73 million dollars. From 2010 to 2017, China\u2019s investment in Syria dropped from 8.12 million dollars to 0.53 million dollars, which led the geo-economic linkage intensity between China and Syria to decrease from 140.14 million dollars in 2006 to 18.96 million dollars in 2017. In particular, it is worth noting that the \u201cMatthew effect\u201d of China\u2019s geo-economic linkage with MSRCs is significant. From 2006 to 2017, the geo-economic linkage intensities of Japan and South Korea increased by 77.80% and 215.17%, respectively, and the geo-economic linkage intensities of Sudan, Yemen, and Syria with China decreased by 17.57%, 41.57%, and 85.71%.The geo-economic linkages between China and other countries in the region are far less intense than those with regional powers such as Japan, South Korea, India, and Saudi Arabia . Taking The interactions of geo-economic flow and the differences in the elements of geo-economic flow cause the different types of geo-economic relations to show different characteristics. Geo-economic linkage intensity can reveal how close the relations between China and the MSRCs are, but it cannot reveal the core characteristics of the geo-economic relations. Therefore, we used the geo-economic linkage intensity as the explained variable, the geo-economic type indices as the explanatory variable, and extreme random forest regression to classify the types of geo-economic relations between China and the MSRCs.Competition and cooperation are the most basic paradigms of geo-economic relations, and their deep-rooted influences cause the studies of types of geo-economic relations to rely on obvious cooperative and competitive tendencies. In earlier research, the logic of competition and cooperation gradually evolved into various forms, such as symbiosis, complementarity, competition, games, and hostility . The geoIn fact, the dominant elements of geo-economic relations include not only distance, politics, market, and resources, but also geo-culture and industrial structure. However, these are not the standard for the classification of geo-relation types in this study, primarily for the following reasons. First, the East Asian civilization centered on China has limited influence on the Maritime Silk Road countries, and its scope mainly extends to Japan, South Korea, North Korea, and Southeast Asian countries, and thus geo-culture cannot be used as a basis for judging the types of geo-economic relations with the MSRCs. Second, the geo-economy is affected by industrial structure complementation and industrial transfer, but China\u2019s current industrial structure adjustment and optimization are still continuing, resulting in a weak contribution to geo-economic development. In addition, relevant studies have shown that geo-economic relations are significantly affected by elements such as the market, geographic distance, and geopolitics . China ahttps://www.mfa.gov.cn/web/ziliao_674904/tytj_674911/tyfg_674913/, accessed on 3 October 2022), we collect 110 Belt and Road cooperation memorandums, trade agreements, investment agreements, economic cooperation framework agreements, technical cooperation agreements, oil and gas cooperation agreements, and assistance agreements, which are signed by the government of the People\u2019s Republic of China, the national development and reform commission of the People\u2019s Republic of China, the Ministry of Foreign Affairs of the People\u2019s Republic of China, the Ministry of Commerce of the People\u2019s Republic of China, and MSRCs. According to the signing of geo-economic cooperation documents, we apply a Likert scale to give scores to geo-economic cooperation documents. The size of a country\u2019s market is closely related to its GDP, per capita GDP, and population. The larger the GDP, the higher the GDP per capita, and the larger the population, the larger the country\u2019s market size. The natural resource endowments of various countries vary greatly, and the spatial distributions of minerals and energy are even more heterogeneous. The total rent of natural resources is the sum of the rents of energy, minerals, and forest resources, reflecting the abundance of a country\u2019s natural resource endowments. The exportation of fuel, ore, and metal reflects a country\u2019s mineral exports. China is a major importer of oil and natural gas, and the oil and gas exported by a country to China can thus reflect the role of oil and gas resources in its geo-economic relations with China. Therefore, we select the fuel exports, total rents of natural resources,, ore and metal exports, and the value of oil and gas exported to China as indices to measure the dominant factors of resource.The distance index usually includes the distance between the capitals and whether the borders are adjacent. On the other hand, the development of modern transportation technology makes transportation time an important index of the influence of geographic distance. Since the economic activities of China and the MSRCs mainly rely on shipping, we select the distance between capitals, whether there is a common boundary, and shipping time as the indices to measure the dominant factors of distance. The cooperative goldenstein factor and conflict goldenstein factor reveal the degrees of geopolitical cooperation and conflict between countries. In view of the important role of geo-economic cooperation documents between China and MSRCs, such as Belt and Road cooperation memorandums, trade agreements, investment agreements, and economic cooperation framework agreements, we take geo-economic cooperation documents as supplementary indexes of dominant geopolitical factors. By searching the treaty documents on the website of the Ministry of Foreign Affairs of the People\u2019s Republic of China is the most important type of geo-economic relation between China and the MSRCs. Among the 39 MSRCs, 21 are market-oriented geo-economic relations, accounting for 53.85%. From a spatial point of view, the market-oriented type is distributed in East Asia, Southeast Asia, South Asia, West Asia, the Balkans, and Africa . South KCountries which are resource-oriented (R) are Yemen, Myanmar, and Kuwait . Kuwait The market-oriented and resource-oriented (M-R) relations are with Vietnam, Brunei, Qatar, Bahrain, Sri Lanka, and Cyprus . VietnamThe geo-economic relations between China and Japan, the Philippines, Iran, Turkey, Pakistan, Egypt, and Slovenia are dominated by market-political-oriented (M-P) means. These countries have each built a relatively complete market economy system and have a close market linkage with China. Among these, Japan and Slovenia have developed economies and high consumption levels, whereas the Philippines, Iran, Turkey, Pakistan, and Egypt have larger populations and larger markets. The importance levels of the conflict goldenstein factor in Japan, the Philippines, and Turkey are 0.21, 0.05, and 0.22, respectively. Geopolical events such as the China\u2013Japan Diaoyu Islands dispute, the China\u2013Philippines Huangyan Island dispute, and Turkey\u2019s support for \"East Turkistan\" terrorists seriously affected their geo-economic interactions with China. China has close geopolitical relations with Pakistan, Iran, Egypt, and Slovenia, and geopolitical cooperation has greatly promoted bilateral geo-economic linkages. China and Pakistan are \u201ciron brothers\u201d with close geopolitical relations, and the China\u2013Pakistan Economic Corridor has become a model of geo-economic cooperation under the framework of the Belt and Road. China and Iran have established a comprehensive strategic partnership, and political mutual trust has promoted the continuous deepening of cooperation between the two countries in various fields, such as energy, trade, and transportation. As the fulcrum country of the BRI in Africa, Egypt has linked its development strategic plan with the construction of the Belt and Road, which promoted the joint construction of the Suez Economic and Trade Cooperation Zone and the New Administrative Capital. Slovenia has responded actively to the BRI, as a result of which China and Slovenia have signed a number of cooperation documents, such as a memorandum of understanding on jointly building the BRI, which has promoted the continuous growth of bilateral economic and trade investment cooperation. It is worth noting that the market is more sensitive to geopolitics, the market-politics-oriented type of geo-economic relations have significant fragility, and geopolitical conflicts can easily lead to a worsening of such geo-economic relations.Only Syria and Greece are resource-dominant and politically dominant (R-P) countries . Syria iOn the whole, the market-oriented type is the most important type of geo-economic relation between China and the MSRCs, and the market-politics-oriented type is the second-most-important type of geo-economic relation. The market-resource-oriented, resource-oriented, and resource- and politics-oriented types are scattered. In the era of geo-economics in the context of globalization, the market economic system has long become the most important economic system in the world. The geo-economic activities of various countries are no longer restricted to the scope of geopolitics, but activities of trade, investment, and geo-economics are carried out under the influence of the market. The multinational operation of enterprises is oriented toward seeking market profits, and geo-economic contacts among countries take economic benefits as an important goal. In geo-economic activities, the position and power of the market are stronger than ever before, making it the dominant force in geo-economic linkages. The nation is the most important subject of geo-economic relations, which determines that geopolitical relations will inevitably affect geo-economic activities. In the geo-economic ties between China and the MSRCs, overseas aid projects, infrastructure connectivity, and economic corridor construction promoted by the country as the mainstays have become important ways to deepen geo-economic linkages. At the same time, major geopolitical conflicts have led to serious setbacks in the geo-economic relations between China and countries involved in geopolitical conflicts. Resource-rich countries have the resources needed for the economic development of other countries, and exporting superior resources is conducive to promoting geo-economic linkages. When resource exports become the most important way for a country to develop its geo-economy, it becomes the main force shaping the country\u2019s geo-economic relations. For economically underdeveloped countries, exporting superior resources or developing resources is the backbone of their national economy, and their external geo-economic relations depend to a large extent on the export of resources. Obviously, the cross-border flow of resources is of great significance to the geo-economic linkages of some countries. The market, resources, and politics do not work in isolation, but also work together to encourage markets, resources, and politics to jointly maintain geo-economic relations.In the context of geo-economic globalization and the reality of major changes unseen in a century, maintaining and handling the geo-economic relations between China and the countries along the Belt and Road are China\u2019s realistic needs. This paper analyzed the types of geo-economic relations between China and the MSRCs, and found that the geo-economic linkage intensity between China and the MSRCs has continued to increase, whose growth phase characteristics are significant. The unbalanced development of investment and trade between China and MSRCs hinders the enhancement of the strength of bilateral geo-economic linkages. China has strong geo-economic linkages with Japan, South Korea, and nine Southeast Asian countries; and regional powers generally have an important geo-economic status. The \u201cMatthew effect\u201d of China\u2019s geo-economic linkage with MSRCs is significant. The classification of geo-economic relation types should go beyond the logic of competition and cooperation and analyze the multiple attributes of geo-economic relations with various influential factors, such as geographic location, geopolitics, market demand, and resource endowments. There are obvious differences in the dominant factors affecting the types of geo-economic relations between China and MSRCs, and the distribution of the importance of the indices of the types of geo-economic relations in each country is disordered. Distance plays a very weak role in the geo-economic relations between China and the MSRCs, but plays an important role in geopolitics, markets, and resources. There are five types of geo-economic relations between China and the MSRCs: market-oriented, resource-oriented, market-resource-oriented, market-geopolitics-oriented, and resource-geopolitics-oriented, of which market-oriented is the most important type of geo-economic relations.It is of great guiding significance to analyze the characteristics of geo-economic linkages and types of geo-economic relations between China and MSRCs for promoting the geo-economic cooperation between China and MSRCs. In order to further promote the development of geo-economic relations between China and MSRCs, China should adopt the following important policies.(1)China should focus on regional powers along the Maritime Silk Road in bilateral geo-economic cooperation. The importance of MSRCs to China\u2019s geo-economic development is not indiscriminate, and regional powers have a greater say in regional geo-economic affairs. Regional powers such as Japan, South Korea, and India occupy important positions in the regional geo-economic development; and Singapore, Pakistan, Egypt, and other countries are the hubs of the shipping channels of the Maritime Silk Road, which are of great significance to China\u2019s construction of the 21st-Century Maritime Silk Road. Therefore, China should be guided by regional powers in its bilateral geo-economic cooperation with MSRCs, maximize the advantage of its economic scale, and continue to promote the steady growth of trade and investment between itself and the regional powers along the Maritime Silk Road. At the same time, China should improve the geo-economic cooperation mechanisms and strengthen political and economic cooperation with countries along the Maritime Silk Road as transportation hubs.(2)Strengthening geopolitical cooperation with MSRCs. The importance of geo-economic relation type indices indicates that geopolitics indices have a strong effect on geo-economic relations. Although the construction of the 21st-Century Maritime Silk Road has been actively supported by many countries, regional powers such as Japan, India, and Turkey have not signed an inter-governmental document on jointly building the Belt and Road with China, and suspicion and vigilance about the construction of China\u2019s Maritime Silk Road still continue. Therefore, China should strengthen geopolitical exchanges with MSRC, reduce or eliminate geopolitical suspicions, and strengthen geopolitical cooperation.(3)Formulating cooperation plans according to the types of geo-economic relations. Due to the differences in geo-economic elements, there are different types of geo-economic relations between China and MSRC, and the cooperation demands of different geo-economic relation types are different. In deepening geo-economic cooperation with MSRC, China should formulate cooperation plans according to geo-economic relation types, take into account the characteristics of the geo-economic development of the cooperative countries, improve the degree of compatibility of geo-economic cooperation between the two sides, and promote potential geo-economic cooperation in practice.In the era led by the \u201cflow space\u201d theory of geo-economics, geo-economic relations have obvious attributes of flow. The interactions of geo-economic flows and the differences in their dominant elements lead different types of geo-economic relations to show different characteristics. Compared with related studies that emphasize the competition and cooperation of geo-economic relations , we beli"} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "We present a database on Brazilian spatial, demographic, and socioeconomic characteristics from 1996 to 2020. This database aims for integration and harmonization with epidemiological data from two major studies. It can also be a valuable database for designing and conducting various types of epidemiologic research, such as health inequality studies, ecological studies (mapping and time-trends), and multi-level analysis.The database gathers official information obtained via open sources from the Brazilian Institute of Geography and Statistics, the Institute for Applied Economic Research, and the Ministry of Health. It includes 139,153 observations and 26 attributes aggregated by years and policy-relevant geographic units on geocoding of municipality centroids, total population size, child population by age-group, birth and mortality measures, Brazilian Municipal Human Development Index, Gini coefficient, Gross Domestic Product, and sanitation. We automated all data processing and curation in the free and open software R. Spatial, demographic, and socioeconomic information is crucial for research, planning, and policy development in health and other sectors. It helps countries compute many health indicators, optimize budgeting and resources allocation, measure and track progress toward international goals and national priorities, and support effective decision-making , 2. BrazWe present a database on Brazilian spatial, demographic, and socioeconomic characteristics from 1996 to 2020. This database aims for integration and harmonization with epidemiological data from two major studies , 5, inclThe database gathers official information obtained via open sources from the Brazilian Institute of Geography and Statistics (IBGE) , 8, the Table The data workflow comprises two main steps. The first step covered the extraction, transformation, and loading routines of data obtained from primary sources of information. The data extraction resulted in 1452 raw files, including spatial data of the Brazilian municipalities, individual data on births and deaths, and aggregated data on population size and socioeconomic characteristics. The key features of data transformation were (i) variables selection/renaming and observations filtering, (ii) calculation of municipality centroids, (iii) correction of codes and names identifying geographic units, (iv) cleansing numeric values, e.g., excluding special characters, and (v) enrichment of the municipal datasets with data aggregated by states, macroregions, and country. This step produced five datasets treated and usable in the database construction.The second step in the workflow involved data integration, harmonization, and enrichment. The IBGE treated-dataset defined the final database structure, in which we combined the other treated datasets according to the years and codes of geographic units. As socioeconomic data was not available for all time points, we applied a simple imputation method for missing data using the next or previous observation of the geographic units. Furthermore, we created the following variables: mortality rate, infant mortality rate, birth rate, estimated population of children under 1-year-old and 1-year-old. The number of children by age group considered two business rules. For children under 1-year-old, we used the MoH estimates in 1996\u20132005 and the number of live births in 2006\u20132020. For children of 1-year-old, we used the MoH estimates in 1996\u20132005 and our estimates in 2006\u20132020 . R codes and data processing/curation were peer-reviewed, and their results compared to the information presented on official sites.We should mention the potential limitations and warnings of the database. First, our eight socioeconomic indicators have different timeframes because of their availability at the municipal level\u2014GDP total and per capita from 1999 to 2018. MHDI , Gini coefficient, and sanitation only 1991, 2000, 2010. It\u2019s worth noting that Brazilian National Household Sample Survey provides some of these indicators for capitals, states, macroregions, and Brazil with a longer timeframe. Moreover, we adopted a simple imputation method for missing data, with several intrinsic limitations, and we presented GDP indicators in Brazilian reais and unadjusted for purchasing power parity. Second, total population size came from the results of demographic censuses , inter-census counts , and population estimates (other years), the only ways to capture these data at the municipal level. Our results for states, macroregions, and Brazil may diverge somewhat from population projections, which do not incorporate post-baseline territorial boundary updates. Finally, the Live Birth Information System (SINASC) and the Mortality Information System (SIM), used to collect live births and deaths data, have variable coverages over time and across geographic units\u2014i.e., lower at the beginning of historical series and underserved areas. Nevertheless, overall SINASC and SIM coverages are high\u201498% and 96%, respectively ."} +{"text": "Correction: BMC Palliative Care 21, 141 (2022)https://doi.org/10.1186/s12904-022-01028-wUpon publication of the original article , an erroThe acknowledgement of our article published in BMC Palliative Care has been incorrect with respect to the contributions of Piret Paal. Piret Paal contributed to the revision/correction of the final manuscript and not to the writing of the manuscript, as the article manuscript was drafted and written only by Nwabata Oji.The corrected acknowledgement is presented here:Data collection on-site, transcription, and qualitative data analysis were conducted by NO. NO drafted the manuscript. The study was conducted in association with TO and OS, who assisted NO in identifying and contacting potential study participants on-site and offered valuable background information on PC in Nigeria. TO and OS revised the content and linguistics of the manuscript. PP supported this research idea and the study conceptually and methodologically, and contributed to the correction of the final manuscript. FE was the research initiator and designed the study. FE conceptually and methodologically supported NO throughout the study. FE also revised the manuscript draft. The qualitative data analysis and interpretation, performed by NO, were discussed continuously and intensively with TO, OS, PP and FE. All authors have read and approved the final manuscript.Another minor error was noticed upon publication of the article.Instead of:physicians\u2019 in practising palliative care\u2014a qualitative study\u201d\u201cPerspectives, perceived self-efficacy, and preparedness of newly qualified The Title should read:physicians in practising palliative care\u2014a qualitative study\u201d\u201cPerspectives, perceived self-efficacy, and preparedness of newly qualified"} +{"text": "Nature Communications 10.1038/s41467-022-29697-4, published online 02 May 2022Correction to: The original version of this Article omitted from the author list the 6th author, Luisa Mercado, who is from the Department of Biomedical Informatics, Harvard Medical School. This has been corrected in both the PDF and HTML versions of the Article. This author was already included in the Author Contributions as L.M. and therefore no further corrections to this statement were made."} +{"text": "Due to extreme weather phenomena, precipitation-induced flooding has become a frequent, widespread, and destructive natural disaster. Risk assessments of flooding have thus become a popular area of research. In this study, we studied the severe precipitation-induced flooding that occurred in Zhengzhou, Henan Province, China, in July 2021. We identified 16 basic indicators, and the random forest algorithm was used to determine the contribution of each indicator to the Zhengzhou flood. We then optimised the selected indicators and introduced the XGBoost algorithm to construct a risk index assessment model of precipitation-induced flooding. Our results identified four primary indicators for precipitation-induced flooding in the study area: total rainfall for three consecutive days, extreme daily rainfall, vegetation cover, and the river system. The Zhengzhou storm and flood risk evaluation model was constructed from 12 indicators: elevation, slope, water system index, extreme daily rainfall, total rainfall for three consecutive days, night-time light brightness, land-use type, proportion of arable land area, gross regional product, proportion of elderly population, vegetation cover, and medical rescue capacity. After streamlining the bottom four indicators in terms of contribution rate, it had the best performance, with an accuracy rate reaching 91.3%. Very high-risk and high-risk areas accounted for 11.46% and 27.50% of the total area of Zhengzhou, respectively, and their distribution was more significantly influenced by the extent of heavy rainfall, direction of river systems, and land types; the medium-risk area was the largest, accounting for 33.96% of the total area; the second-lowest-risk and low-risk areas together accounted for 27.09%. The areas with the highest risk of heavy rainfall and flooding in Zhengzhou were in the Erqi, Guanchenghui, Jinshui, Zhongyuan, and Huizi Districts and the western part of Xinmi City; these areas should be given priority attention during disaster monitoring and early warning and risk prevention and control. Precipitation-induced flooding refers to the intense accumulation of precipitation, rising or overflowing water levels in rivers, lakes, and reservoirs, and the inability of water to drain away due to heavy or continuous rainfall ; this caFrom 19 to 23 July 2021, Henan Province in central China experienced anomalously heavy rainfall, with precipitation of >400 mm at 43 observation stations, >300 mm at 154 stations, >200 mm at 467 stations, and >100 mm at 1426 stations. During that time, 19 cities and counties in the province broke their daily precipitation records. Zhengzhou, the provincial capital, which is located in a flood disaster zone, experienced a total rainfall of 993.1 mm and a cumulative surface rainfall of 543 mm during the event , high-risk areas , moderate-risk areas , low-risk areas , and very low-risk areas . The spatial distributions of these five levels are shown in 2, accounting for 11.46% of the city\u2019s area. These are mainly in Erqi District, Guancheng Hui District, Jinshui District, Zhongyuan District, Huiji District, and the western part of Xinmi City as well as the northern part of Xingyang City near the Yellow River system. High-risk areas total 2081.18 km2 and account for 27.5% of the city\u2019s total area. They are located around the very high-risk areas, mainly in Xinmi City in the central area, the northern parts of Xingyang City and Gongyi City, and the eastern part of Dengfeng City. Areas at moderate risk account for the largest area at 2569.64 km2, which is 33.96% of the city\u2019s total area. They are the most widely distributed, including the 11 districts, counties, and county-level cities of Dengfeng City, Gongyi City, Shangjie District, Xingyang City, Xinmi City, Xinzheng City, Zhongmu County, Guancheng Hui District, Jinshui District, Zhongyuan District, and Huiji District. The spatial distribution of moderate-risk areas is strongly coupled with geographic factors, including elevation, slope angle, and land-use type. Low-risk and very low-risk areas cover 1292.25 km2 and 757.12 km2, respectively, and together they account for 27.09% of the total area of Zhengzhou. They are mainly in Gongyi City and Dengfeng City in the west of Zhengzhou and Zhongmu County and Xinzheng City in the east.The total area of very high-risk areas in Zhengzhou is 866.81 km(1) The occurrences of storm and flood disasters are not independent events caused solely by sudden and heavy rainfall. Rather, the causes are often intertwined with external factors, including a region\u2019s physical geography and socio-economic situation. Based on the \u201cNatural Disaster Risk System\u201d, this study conducted a census of all factors causing storm and flood disasters, and established a basic database of regional storm and flood disaster risks. The rainstorm and flood risk assessment model in Zhengzhou was effective in accurately assessing disaster risks from multiple perspectives.(2) This study used machine learning algorithms to develop a storm and flood risk assessment system, overcoming traditional problems with indicator selection and weighting calculation processes being affected by the complex non-linear relationship between data and excessive subjective human influence. This work lays a foundation for further research applications of storm and flood risk assessments based on artificial intelligence and big data that will likely be of growing interest for the field of flood risk analysis.(3) Drawing on the advantages of big data, remote sensing, global positioning systems (GPS), and geographical information systems (GIS), this study built on previous research using remote sensing of night-time light as an indicator, replacing the socio-economic factors of population density, urbanisation level, and per capita GDP. In doing so, it overcomes the disadvantage of excessive correlation between traditional socio-economic factors in such studies. At the same time, its grid-based data improved the accuracy of the evaluation results compared to traditional socio-economic factors that use the administrative region as a unit.(1) Using the single variable maps and comprehensive risk assessment and zoning maps of the storm and flood risk in Zhengzhou, each district (city) can be managed according to its risk level.The high-risk areas are mainly distributed in the Erqi, Guancheng Hui, Jinshui, Zhongyuan, and Huiji Districts, and the west of Xinmi City and north of Xingyang City, which are significantly affected by extreme precipitation. For these high-risk areas, cities should prioritise improving urban flood prevention emergency plans, strengthening real-time monitoring of precipitation and real-time evaluation, and improving the mechanism of information distribution regarding storms and floods. Simultaneously, as the populations and economies of these areas are relatively dense, improving public awareness of flood disaster prevention is particularly important . EffortsThe second-highest risk areas included the cities of Xinmi and Xingyang City, the northern part of Gongyi City, and the eastern part of Dengfeng City, which all experience location-specific exposure and flood-aggravating susceptibility.These regions have developed fluvial systems and a high proportion of arable land. Therefore, the focuses for preventing storm and flood disasters should be flood control engineering and urban land-use planning, including: building flood walls and embankments along the river, strengthening existing flood control and drainage infrastructure, and preparing early flood warning and emergency plans. Furthermore, it is also important to strengthen land-use planning, management, and control, prioritise urban flood prevention, and coordinate and adapt urban construction to floods.The eleven medium-risk areas include Zhongmu County; the cities of Dengfeng, Gongyi, Xingyang, Xinmi, and Xinzheng; and the Shangjie, Guancheng Hui, Jinshui, Zhongyuan, and Huiji Districts. These areas are moderately affected by flood-aggravating susceptibility, flood-causing risk, and location-specific exposures, but more importantly, they have a weak ability to prevent and reduce disasters. Therefore, emphasis should be placed on improving regional emergency management capabilities and enhancing public awareness of flood controls. On one hand, it is necessary to formulate emergency plans in large- and medium-sized cities, strengthen the allocation and coordination of manpower, and improve supervision and management to ensure timely early warnings reach every affected individual. On the other hand, the publication and education of meteorological information and emergency response capabilities can be iThe moderately low-risk and low-risk areas are mainly distributed in Zhongmu County, Xinzheng City, and the intersection of Gongyi Dengfeng Cities. Apart from public outreach and education, no other risk management is required.(2) Under global warming, the frequency and intensity of extreme climate disasters have increased significantly for cities that are currently affected. Urban planning and construction are intrinsic to extreme climate risks. Multi-scale theoretical and practical research of disaster mitigation, adaptation, and planning is an important means of mitigating extreme climate disasters and improving urban resilience for the future. Using multiple forms of information technologies to conduct urban storm and flood disaster risk assessments, high-risk areas can be identified and urban flood control and drainage plans developed in advance. This is consistent with the three-step strategy of \u201cassessment\u2013warning\u2013strategy\u201d, and using flood control and risk reduction projects to manage flood disasters. Effectively restricting the construction and urban planning of lower-level sponge cities is beneficial to strengthening the blue and green lines, which represent water bodies and natural systems, management of ecological spaces, and vertical urban management. This method also allows for the optimisation of engineering decision-making, restricting or optimising projects and socio-economic development in risk areas, and improving early warning and forecasting.Using the precipitation-induced flooding event that occurred in the city of Zhengzhou in the central Chinese province of Henan in July 2021 as a case study, we used the RF and XGBoost algorithms to examine the influencing factors and conduct a risk assessment of precipitation-induced flooding in Zhengzhou. Our research led to the following conclusions.(1) Based on our evaluation of the geographical features of Zhengzhou and the quality of available data, we selected 16 indicators from the four aspects of geography, meteorology, population, and economy. We used RF to examine the contribution of each indicator to precipitation-induced flooding. We found that the four indicators with the highest contributions were (in descending order) total rainfall over three consecutive days, extreme daily rainfall, vegetation cover, and river systems. The indicators with the next highest contributions were land-use type, elevation, night-time light brightness, GDP, and slope angle.(2) Based on the AUC and ACC of the RF model, we streamlined the indicators to create an optimised risk-assessment index. After removing the four indicators of economic growth, roads, slope aspect, and per capita GDP, which were the four bottom-ranked indicators in terms of contribution, the model\u2019s prediction accuracy and performance were optimal.(3) We used XGBoost to calculate the weights of the streamlined indicators for the final objective of constructing a risk-assessment model of precipitation-induced flooding in Zhengzhou to individually assess the four aspects of flood-aggravating environmental susceptibility, flood-causing risk, location-specific exposure, and flood-mitigation capability, which were integrated to obtain the final risk-assessment results of precipitation-induced flooding in Zhengzhou. The results showed that very high-risk and high-risk areas account for 11.46% and 27.50% of the total area of Zhengzhou, respectively. Their distribution is significantly affected by heavy rainfall, river systems, and land-use type. The areas with the highest risk of precipitation-induced flooding in Zhengzhou are Erqi District, Guancheng Hui District, Jinshui District, Zhongyuan District, Huiji District, and western Xinmi City.(4) This study innovatively introduced a machine learning algorithm in the construction of a risk-assessment system for flooding; this overcomes the problem that the process of screening index factors and calculating weights is affected by complex non-linear relationships between data and excessive human subjective influence, improving the accuracy of the research results. This study can help government departments to identify high-risk areas and provide a scientific basis for storm and flood prevention and mitigation planning. However, there are still some limitations in this paper: (i) due to the limitation of data acquisition, only 18 basic factors were selected for secondary optimisation and screening, and it is proposed to expand the basic database in multiple ways in future studies and to continue to explore more comprehensive storm and flood hazard impact factors and their mechanism of action; (ii) due to the limitation of the original data type, some of the data were sourced from statistical tables and processed as panel data by the kriging difference method, resulting in the lack of precision of the data. In the future, we may consider and seek other raster data with higher accuracy for similar replacement."} +{"text": "The USA , countries in the European Region , and high-income countries were dominant publishing countries. Of 699 research articles and systematic reviews, surveillance and trends of physical activity were the main research area, followed by health outcomes, and correlates and determinants of physical activity. There is a wide gap in publication productivity in the field of physical activity and health during the pandemic among different countries\u2019 economic statuses.The coronavirus disease 2019 (COVID-19) pandemic induced a sudden surge in COVID-19 related publications. This bibliometric analysis aimed to analyze literature on physical activity and COVID-19 published in the PubMed database. The search terms AND COVID-19 [MeSH Terms]) were applied to obtain publications from the inception of PubMed to February 2022. The analyses included the year of publication, type of publication, and origin of publication by country, region, and country income. The research areas were analyzed for research articles and systematic reviews. Of 1268 articles, 143 articles were excluded, and 1125 articles were analyzed. A total of 709 articles (63.02%) were published in 2021. A majority of publications were research articles ( The gro in 2021 .Since late 2019, a newly emerging disease, coronavirus disease 2019 (COVID-19), has affected several aspects of health, including physical activity ,6,7,8,9.In addition, the COVID-19 pandemic has changed levels and patterns of physical activity ,13. ChanPublished articles were retrieved from PubMed, a database of biomedical and life sciences literature that contained more than 33 million records . The seaThe search results retrieved from PubMed were downloaded as a .csv file to maintain the number of articles on the search date. The .csv file was converted to a .xlsx (a default file format for Microsoft Excel).The article title and year of publication were kept in their original forms (.csv file). Type of publication was classified as (i) research/original article, (ii) systematic review, (iii) editorial, (iv) letter to the editor/research letter, and (v) miscellaneous article. The country of publication was defined as the country of the first author\u2019s affiliation. The countries were grouped according to the World Health Organization (WHO) regions: (i) African Region, (ii) Eastern Mediterranean Region, (iii) European Region, (iv) Region of the Americas, (v) South-East Asia Region, and (vi) Western Pacific Region . The WorThe lead author (A.W.) reviewed the titles and abstracts to identify the physical activity research area for research/original articles and systematic reviews : (i) surveillance and trends, (ii) correlates and determinants, (iii) health outcomes, (iv) interventions and programs, (v) policies , and vi. ArticleEach included article was identified as a type of publication and the country of the first author. The region and income of the country were checked on the WHO and the World Bank\u2019s websites ,19. DataSubsequently, research/original articles and systematic reviews were analyzed qualitatively by the lead author (A.W.) to identify the research areas according to the description in n = 709, 63.02%) were published in 2021, while the rest of articles were published in 2020 and 2022 . Research or original articles were the most common publication type , the European Region , and high-income countries were the most dominant publication productivity . The USAuctivity .n = 416 out of 699 articles) of research/original articles and systematic reviews related to COVID-19 were in the area of surveillance and trends. The second most common research area was health outcomes .A majority of articles related to physical activity and COVID-19 published in the PubMed database were research or original articles (60.27%) and systematic reviews (1.87%). The rest of the publications (37.86%) were classified as non-research articles. Authors affiliated with institutions in the USA, the UK, and Italy contributed more than three in ten publications. More than three-quarters of publications were produced by authors in high-income countries. In contrast, there was no publication from authors in low-income countries. Countries from the European and Americas regions contributed to nearly three-quarters of the publications. Most research articles and systematic reviews focused on surveillance and trends, health outcomes, and correlates and determinants of physical activity during the COVID-19 pandemic.This study found approximately four in ten publications relevant to physical activity and COVID-19 were published as non-research articles by February 2022. The proportion of non-research articles was higher compared to the ratio of research and non-research articles in other fields . A studyOur study presented the top ten dominant countries in physical activity and health publications. Of ten countries, eight are classified as high-income countries (except that Brazil and China are upper-middle-income countries). This finding was consistent with previous evidence. Fontelo et al. analyzed the publication trend in PubMed from 1995 to 2015 and identified the top 30 publishing countries . The topIn terms of country income, our findings were in line with a study by Ram\u00edrez Varela et al. that higher-income countries produced more publications on physical activity and publications per capita from 1950 to 2019 ,25,26. AWe identified the five main areas of physical activity research according to the expert excerpts . HoweverOur findings revealed some observations and recommendations for scholars and future research. A large proportion of non-research articles were published in the form of expert excerpts. Although these pieces of literature raised valuable knowledge, the knowledge of and perspectives toward COVID-19 were dynamic, especially in the early phase of the outbreak ,31. TherThere were three major strengths of this study. First, the search strategy was inclusive. We used the MeSH terms of both ends of the spectrum of physical activity and COVID-19 to obtain the variety of terms used in articles. Second, we excluded non-relevant articles based on the exclusion criteria prior to the analysis. Some published bibliometric analyses included all search results for the analysis. We also excluded some articles that contained keywords relating to physical activity in their titles ,33, howeDespite the strengths of the study, there are certain limitations worth noting. First, we indicated the type of publication based on article types assigned by the journals. However, some journals considered systematic reviews as review articles, while others classified these as research/original articles. We assigned these articles following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline with/without the systematic review registration as \u201csystematic review\u201d in our analysis. Second, the country of the first author\u2019s affiliation might not represent the site of the study, as the first author may conduct the study in another country or several countries. Our analysis cited only accounts for the affiliating country of the first author. Third, this study did not include the analyses of keywords, author names, and affiliation names.This bibliometric analysis disclosed the characteristics and trends of the literature on physical activity and COVID-19 published in PubMed up to February 2022. A majority of articles were research or original articles, while nearly 40 percent were classified as non-research articles . The USA, European countries and high-income countries were dominant publishing countries of publications on physical activity and health during the COVID-19 pandemic. There was no publication by lead authors affiliated with institutions in low-income countries. This reflected the impact of the country\u2019s income on the publication productivity in this field. More than half of the research articles and systematic reviews involved the research area of surveillance and trends of physical activity. Physical activity-related policies were the least common area among research articles and systematic reviews. Building research capacity and supporting the mechanisms to drive research productivity in low- and middle-income countries are required to improve the global research productivity in the field of physical activity and health."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Correction: BMC Public Health 22, 1240 (2022)https://doi.org/10.1186/s12889-022-13646-3In the original publication there waIncorrect funding106534]. The funding body had no role in the study design and collection, analysis, and interpretation of data and in writing the manuscript.This study was supported by the Wellcome Trust [Correct funding106534/Z14/Z]. The funding body had no role in the study design and collection, analysis, and interpretation of data and in writing the manuscript.This work was supported by the Wellcome Trust ["} +{"text": "Nature Communications 10.1038/s41467-022-31548-1, published online 08 July 2022Correction to: The original version of this Article omitted from the author list the 17th author Feng Gao, who is from the Key Laboratory of Aerospace Medicine of the Ministry of Education, School of Aerospace Medicine, Fourth Military Medical University, Xi\u2019an, 710032, China. This has been corrected in both the PDF and HTML versions of the Article."} +{"text": "The high technical barrier to entry in the field of neuroimaging can hinder early insight from promising results and the development of evidence-based clinical practice.The working group focused on published literature in order to develop a new methodology in the analysis, visualization, and representation of fMRI data in the psychiatric setting.Three valid and established measures were chosen, in order to achieve dimensionality reduction, stability and explainability of results, namely Regional-Homogeneity; fractional Amplitude of Low-Frequency Fluctuations; Eigenvector-Centrality. Each measure was color coded and individual images per subject compiled, averaging results by functional networks as described the FIND lab of the University of Stanford. 272 individual scans were processed .The discriminative power between clinical groups of the novel method was significant both by human eye, and later confirmation by statistical tests, and by computer vision algorithms . The precision-recall Area Under the Curve, dividing by 80/20 proportion between train and test sets, was >84.5% for each group. The group of patients with Bipolar Disorder showed a partial overlap with the group of patients suffering from Schizophrenia \u2013 by a dominance of Eigenvector-Centrality and Regional-Homogeneity, as well as a lower prevalence of fractional Amplitude of Low-Frequency Fluctuations, for both in comparison to controls.The present study offers preliminary evidence for the adoption of i-ECO (integrated-Explainability through Color Coding) in fMRI analyses during rest in the Psychiatric field.No significant relationships."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "The Special Issue \u201cSynthesis, Functionalization and Applications of Nanocarbons\u201d starts from the growing interest of the scientific community in carbon-based materials and the various applications of these versatile compounds. Among the first allotropic forms of carbon that strongly attracted the attention of scientists, there are the fullerenes; then, several works were produced with carbon nanotubes; and, in 2010, studies about graphene earned the Nobel prize for Andre Geim and Konstantin Novoselov. More recently, other carbon-based materials such as nanodiamonds, onion-like carbon nanomaterials, carbon dots, and carbon quantum dots were added to the large and fascinating family of carbon-based nanomaterials.The Special Issue presents seven contributions focused on carbon-based nanostructures in the form of graphene, carbon dots (CDs), nanodiamonds, onion-like carbon nanomaterials and carbonous doped materials. The applications of nanocarbons include wearable devices, water remediation, energetic applications and the fabrication of sensing devices. The use of carbon nanostructures in wearable devices allowed the fabrication of highly stretchable materials, preserving their electric properties and ensuring high biocompatibility . Co- andAn environmentally sustainable procedure to obtain graphene oxide starting from graphene was proposed by C. F. Costa and collaborators . An alteFinally, a research article and a fascinating review concerning a new emerging class of carbon-derived materials, carbon dots, are presented in this Special Issue ,7. Sever"} +{"text": "Joseph Pratt, a sanatorium doctor, at the beginning of the 20th century began to organize groups of patients in order to transmit information about their illness, observing that those who came had a better evolution. In the twenties, Jacob L. Moreno, would make the leap towards mental health, transferring the group format to the treatment of mental disorders. At the same time, Lazell and Marsh began to carry out psychoeducational groups with admitted schizophrenic patients.Present experience of a psychotherapeutic group in a brief psychiatry hospitalization unit.Non-directional, voluntary group, with daily frequency and 30 minutes duration. Between 8-15 patients participated. Participation in the group required compliance with 2 rules: respecting word turns and speaking from one\u2019s own experience. The sessions were organized in three parts, 1. Opening of the group: the rules are remembered and we welcome new patients. 2. Group: dialogue between patients 3. Group closure: summary of the session and dismissal of discharge patients.The following topics were addressed: - The experience of admission; traumatic vs restorative. - The difficulties they expected to encounter after discharge. - Aspects related to family bonding, between equals and couples. As difficulties we find: - The heterogeneity in the symptoms of the patients. - Voluntary participation in the group. - Conflicts reactive to non-compliance with the rules.Group therapies in brief hospitalization units have great therapeutic potential.No significant relationships."} +{"text": "PLOS Medicine, together with guest editors Timothy Walsh, Ramanan Laxminarayan and Ana Cristina Gales, announce a forthcoming Special Issue dedicated to bacterial antimicrobial resistance (AMR). Research submissions are now being invited.The editors of The emergence of pathogenic bacteria which cannot be effectively treated with existing drugs has been prioritised by the World Health Organization as one of the top ten global public health threats facing humanity . Drug-reAMR is a One Health problem and its causes lie in human, animal and environmental domains. The overuse and misuse of antibiotics, and the potential for transmission within and between these domains is responsible for the rapid global spread of drug-resistant pathogens. Use of antibiotics increased by 65% globally between 2000 and 2015, and more than doubled in LMICs over the same period . PathogePLOS Medicine editors seek high-quality and high-impact research submissions related to the main drivers, surveillance and prevention of bacterial antimicrobial resistance, particularly in low- and middle-income settings. Areas of particular interest include the prevalence and clinical challenges of drug-resistant bacteria, interventions to reduce disease transmission, diagnostics informing antimicrobial prescribing, misuse and overuse of antimicrobials, economics of antimicrobial access and use, and One Health interventions to reduce AMR. Submission of articles related to pathogens of highest concern and highest global burden (excluding Mycobacterium tuberculosis) are strongly encouraged.The guest editors and plos.io/AMR for more detailed information.Please see http://journals.plos.org/plosmedicine/s/submit-now, indicating your interest in the Special Issue in your cover letter. Questions about the Special Issue can be directed to plosmedicine@plos.org.To submit your manuscript for consideration, please visit th 2022.The submission deadline is July 15"} +{"text": "Diagnosis and treatment of drug-resistant tuberculosis (DR-TB) have radically changed in accordance with recommendations from the World Health Organization (WHO) in the past decade, allowing rapid and simple diagnosis and shorter treatment duration with new and repurposed drugs.A descriptive analysis of the status and progress of DR-TB diagnosis and treatment in six priority countries in the Western Pacific Region was conducted using information from interviews with countries and the WHO TB database.Over the past decade, the use of Xpert MTB/RIF has increased in the six priority countries, in parallel with implementation of national policies and algorithms to use Xpert MTB/RIF as an initial diagnostic test for TB and detection of rifampicin resistance. This has resulted in increases in the number of people diagnosed with multidrug-resistant or rifampicin-resistant TB (MDR/RR-TB). Shorter treatment regimens with new and repurposed drugs have also been adopted for MDR/RR-TB cases, alongside a decentralized model of care, leading to improved treatment outcomes.The Western Pacific Region has achieved considerable progress in the diagnosis and treatment of DR-TB, in line with the evolving WHO recommendations in the past decade. The continued commitment of Member States is needed to address remaining challenges, such as the impact of the coronavirus disease pandemic, suboptimal management and health system issues. Tuberculosis (TB) continues to be a major global health challenge. Despite a concerted global effort to eliminate it, TB remains one of the leading infectious causes of death globally. In 2020, an estimated 10 million incident cases of TB and 1.5 million TB-related deaths occurred worldwide. and Xpert MTB/XDR for diagnostic tests;use of shorter all-oral bedaquiline-containing regimens;discontinuation of kanamycin and capreomycin for MDR/RR-TB; anduse of a bedaquiline, pretomanid and linezolid (BPaL) regimen. ;percentage of MDR/RR-TB cases tested for susceptibility to fluoroquinolones;number of MDR/RR-TB cases treated with bedaquiline, shorter regimen and all-oral longer regimen; andtreatment outcomes for MDR/RR-TB cases started on treatment. .All data analyses and visualizations were conducted using the statistical software package R 4.1.1 has been introduced in all priority countries. In Cambodia, Xpert Ultra had already replaced Xpert MTB/RIF at the time of the interview. In the other priority countries, Xpert Ultra is being used together with Xpert MTB/RIF. There are also plans to introduce Xpert MTB/XDR in 2022 in the Lao People's Democratic Republic, Papua New Guinea, the Philippines and Viet Nam.In Cambodia, first-line line probe assays (FL-LPAs) are used to detect isoniazid resistance among rifampicin-susceptible TB cases only on an ad hoc basis. However, in Mongolia, Papua New Guinea and the Philippines, FL-LPAs are routinely used to detect isoniazid resistance among rifampicin-resistant TB cases, and in Cambodia and Viet Nam, they are used in such cases but on an ad hoc basis. Second-line line probe assays (SL-LPAs) are used as an initial test to detect fluoroquinolone resistance among confirmed MDR/RR-TB cases in all priority countries. The use of Truenat (a point-of-care rapid molecular test) for detection of TB and rifampicin resistance is planned as a pilot project in 2022 in Cambodia, the Philippines and Viet Nam.As the use of new and repurposed drugs in shorter and longer regimens is scaled up in priority countries, phenotypic drug susceptibility testing for those drugs is conducted or planned. In the Lao People's Democratic Republic, drug susceptibility testing for linezolid is in place, and drug susceptibility testing for bedaquiline, clofazimine and delamanid is planned. In Mongolia, drug susceptibility testing for bedaquiline, linezolid and clofazimine is conducted. In Viet Nam, drug susceptibility testing for bedaquiline, linezolid, clofazimine and delamanid is conducted and drug susceptibility testing for pretomanid is planned.Fig.\u00a01). The number of people diagnosed with MDR/RR-TB and the number starting on treatment per year in all countries has increased since 2010 . However, in the Lao People's Democratic Republic, Mongolia and the Philippines, there was a decrease in the number of MDR/RR-TB cases diagnosed in 2020 compared with 2019. This decrease started in 2014 in Mongolia. The proportion of enrolment in treatment among diagnosed cases exceeded 80% in 2020 in Mongolia (104%) and Viet Nam (89%), whereas it was 80% or below in the Lao People's Democratic Republic (80%), Papua New Guinea (76%) and the Philippines (78%).The percentage of new and previously treated TB patients tested for rifampicin resistance in four of the priority countries (no reports from Cambodia and Papua New Guinea) increased between 2017 and 2020, with some fluctuations .The percentage of MDR/RR-TB cases tested for susceptibility to fluoroquinolones in four priority countries between 2015 and 2020 (no reports from Cambodia and Papua New Guinea) varied with increased coverage in Mongolia and Viet Nam alongside a recommended MDR-TB regimen is being undertaken in Mongolia and the Philippines.Various treatment adherence interventions for MDR/RR-TB patients are offered in the six priority countries by health-care workers or family members. In Mongolia, Papua New Guinea and the Philippines, both facility-based DOT by health-care workers and community-based DOT by family members are used. In the Lao People's Democratic Republic, the main modality is facility-based DOT by health-care workers during hospitalization. Video-observed treatment (VOT) has been conducted as a pilot project in Mongolia, the Philippines and Viet Nam.MDR/RR-TB patients are treated mainly through ambulatory care in Papua New Guinea and the Philippines, whereas most patients are treated during hospitalization in the Lao People's Democratic Republic. In Mongolia, patients are hospitalized until their sputum conversion from positive to negative, and in Viet Nam, patients are hospitalized for up to 1\u00a0month. In Cambodia, patients were hospitalized for the first week for a workup and monitoring of a new regimen before the coronavirus disease (COVID-19) pandemic; however, this practice has since been restricted to ambulatory care only.Fig.\u00a04). The use of the all-oral longer regimen increased between 2019 and 2020 in the Philippines (no report from the other countries).Where data were available, the number of MDR/RR-TB cases treated with bedaquiline and the shorter regimen between 2015 and 2019 increased in all six priority countries, although there were decreases observed between 2018 and 2019 in Cambodia, the Lao People's Democratic Republic and the Philippines . The proportion of cases with treatment success increased in the Lao People's Democratic Republic (from 67% in 2014 to 84% in 2018) and the Philippines (from 46% in 2014 to 67% in 2018), mainly due to a reduction in the proportion of treatment failure in the Lao People's Democratic Republic and to patient loss to follow-up in the Philippines. The proportion of cases with treatment success was similar each year in Mongolia and Viet Nam, and fluctuated from year to year in Cambodia and Papua New Guinea.Treatment outcomes for MDR/RR-TB cases started on treatment in 2014\u20132018 in the six priority countries differed , given the high rate of isoniazid resistance in some countries. Moreover, the regimen for Hr-TB should be implemented in more of the priority countries.There was static and fluctuating MDR/RR-TB treatment success in some priority countries, despite the roll-out of shorter regimens. Although they are shorter, these regimens do not guarantee improved treatment outcomes because they still require clinical management, strong patient support and monitoring systems to ensure patient adherence to treatment. In those countries, expansion of shorter regimens should be reinforced by optimal management and supportive health systems for improved treatment outcomes.Our analysis has several limitations. First, there were no reported data for some indicators for certain years or from particular countries in the WHO database, impeding a complete analysis. Second, there is a possibility of recall bias from PMDT focal points, despite verification by follow-up communication or subsequent rGLC missions. Although attempts were made to refer to official documents, some answers were provided based on memory. Notwithstanding these limitations, this analysis provides a comprehensive and practical insight into the progress of PMDT in these six priority countries in the region.In conclusion, these six priority countries in the Western Pacific Region, in collaboration with the rGLC, have achieved considerable progress in the diagnosis and treatment of DR-TB in line with the evolving WHO recommendations over the past decade. Automated nucleic acid amplification tests and shorter all-oral regimens containing new and repurposed drugs are now used for DR-TB diagnosis and treatment in the region, leading to reductions in the case-detection gap and enhanced treatment outcomes. However, several challenges remain, particularly the impact of the COVID-19 pandemic, suboptimal patient management and health system issues. The continued commitment of countries to a speedy recovery from COVID-19, patient-centred care, capacity building and a robust health system is needed to continue progressing towards ending DR-TB in the region."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Schizothorax eurystomus, Kessler 1872 is a unique economic fish in Xinjiang, China that is rarely seen in the market. Next-generation sequencing (NGS) was used to determine the complete mitochondrial genome of S. eurystomus collected from the Yarkand River in Xinjiang. The results showed that the mitochondrial genome is a circular, 16,488-bp-long nucleotide with the typical vertebrate genome structure of 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and a control region. The termination-associated sequence (TAS), central conserved sequence block (CSB), and conserved sequence block were detected in the control region. Phylogenetic analysis placed S. eurystomus in a fully supported clade with S. biddulphi, and that clade was sister to S. yunnanensis. To our knowledge, this is the first study on the complete mitochondrial genome of S. eurystomus from the Yarkand River in Xinjiang, and it provides baseline genetic information for future studies. Schizothorax eurystomus, Kessler 1872, also known as the wide-mouth hip scale fish, is a cold-water white fish that belongs to order Cypriniformes, family Cyprinidae, and subfamily Schizothoracinae has revolutionized the field of molecular biology because it is rapid and can generate large amounts of genomic data Schuster . TherefoS. eurystomus collected from Altash Station of the Xinjiang Yarkand River in September 2021. A specimen was deposited at the CAS Key Laboratory of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences under voucher number L212. The Illumina NovaSeq sequencing platform was used to process sequences.DNA was extracted from muscle tissue of http://mitos2.bioinf.uni-leipzig.de/index.py) . The circular map of the complete S. eurystomus mitochondrial genome is shown in Similar to the size of other teleost mitogenomes, the complete mitochondrial genome of S. eurystomus were encoded on the heavy-strand (H-strand), with only NAD6 and 8 tRNA genes encoded on the light-strand (L-strand) , and GTG was the initiation codon for COI , TAG terminated three genes ; the incomplete T\u2013 terminated the COII, NAD4, and COB genes by post-transcriptional polyadenylation . The ATG for COI . The stoS. eurystomus were 956- and 1630-bp long, respectively. They were located between tRNAPhe and tRNALeu, and were separated by tRNAVal, similar to other vertebrates , central conserved sequence block (CSB), and CSB were detected in the control region and were similar to those of most bony fishes , however, there is no paper published for the description in details on it. In this study, we collected and identified five specimens, and then used NGS to obtain their mitochondrial genome. The complete sequences were used for phylogenetic analysis and the phylogenetic results revealed that a previously collected specimen (KY436758.1) was genetically distant from other related species; therefore, the complete mitochondrial genome of S. eurystomus needed to be updated. Phylogenetic analysis of the complete mitochondrial genome of S. eurystomus revealed that it belonged to a clade with S. biddulphi, and they are sister to S. yunnanensis (S. eurystomus in detail and provides a scientific basis for future molecular systematic and phylogenetic studies of bony fishes in Cyprinidae.\u2003One complete mitogenome of nanensis . This st"} +{"text": "Human-Media Interaction sections, Frontiers in Robotics and AI with its Human-Robot Interaction and Multi-Robot Systems sections, and Frontiers in Artificial Intelligence with its AI for Human Learning and Behavior Change section.Computer system design has been driven predominantly by technical aspects and considerations. However, as computing systems are increasingly embedded and integrated into the world, they have an extended impact on individuals and societies. In this context, social and behavioral dynamics become equally essential components of computing systems research, notably including robotics and artificial intelligence. The increasing shift towards socio-technical systems and the integration of collaborative and cooperative aspects require new multidisciplinary efforts and a focus on the larger system context and evolution. In hybrid systems, i.e., artificial agents, robots, and humans interacting with each other, explicitly considering the underlying social relations and dynamics is integral to the ability to design robust, adaptive, purpose- and useful systems. Recent advances in machine learning and other computational techniques allow for the effective real-time analysis of complex interaction behavior. This holds both for the individual constituents as well as for the integration of subsystems. This highly multidisciplinary Research Topic, therefore, spans various journals including Frontiers in Psychology and Frontiers in Computer Science with their Taking a systemic, cross-disciplinary perspective on human-AI and human-robot interaction, as well as their effects on the larger system context, we encouraged submissions that integrate dedicated findings on socio-technical settings from, e.g., neuroscience, psychology, sociology, economics, and other relevant disciplines, into novel interaction and system design approaches. Original research, reviews, tools, databases, benchmarks, and evaluation methods relative to the following topics were welcome.The international submissions reflect this variety, with the group of accepted manuscripts comprising original research, methods, and review articles covering various perspectives on designing for human-machine systems that form socio-technical ecologies.von Terzi et al. address the problem from a psychological perspective, comparing the fulfillment of needs while experiencing technologies and how that is linked to social context, i.e., private and public settings. The results indicate significant effects for relatedness and popularity, potentially improving systems design.In their article, Raymond et al. analyze the problem of fairness in decentralized conflict resolution and how privacy limitations constitute a trade-off for fairness losses. A series of randomized and application examples compare different strategies and evaluate perspective/scope as the objective global fairness measure against the perceived local fairness.Bagheri et al. look at transparent interaction-based learning mechanisms for human-robot collaboration to improve the training efficiency, efficacy, and performance of collaborative robots (cobots) working with non-expert human partners.Leichtmann et al. review possible replicability issues in human-robot interaction and provide methodological, statistical, and systemic suggestions to harden and improve future studies regarding comparability, reliability, and robustness.Boos et al. introduce a compliance-reactance model to evaluate new human-robot interaction approaches on the system level through behavioral and affective factors. Their framework translates well-known elements from inter-human social interaction to human-robot and other systems, underscoring the importance of providing social cues to successful deployment and social acceptance.Finally, Other submissions covered topics ranging from user perception of privacy, over abstract UI concepts, to questions of presence in collaborative work and communication.As human-machine systems are deployed and becoming a reality in numerous scenarios involving different stakeholders, there is a need to shift system design towards more holistic approaches, which consider second-order effects on the host environment (\u201chabitat\u201d) and, in consequence, on the embedded systems. This explicitly includes long periods of time where systems of systems form ecologies and co-evolve after deployment. Highly interactive settings, such as an advanced smart city scenario, comprise many heterogeneous systems, e.g., robots and/or artificial agents, crowd-sourced data, social media, location-based applications, swarms of delivery drones, cleaning or gardening robots. The goal is to improve urban life, such as transport systems, healthcare, infrastructure, and services, by collecting and analyzing data from a wide range of sensors and applications. However, many of these systems fail to adapt or serve their intended purpose within the larger socio-technical ecosystem as they are oblivious to the complex dynamics in their immediate context, let alone the effects on cities as larger organisms. It is therefore vital to establish an integral way of designing human-machine systems that form socio-technical ecologies, allowing them to respect and adapt to the ever-evolving context in which they are embedded.We believe this Research Topic constitutes a step towards a more comprehensive, multidisciplinary view of human-machine systems and their design. Our thanks go to all reviewers for their in-depth assessments and to the authors for their contributions."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Structural biology is an essential tool for understanding the molecular basis of diseases, which can guide the rational design of new drugs, vaccines, and the optimisation of existing medicines. However, most African countries do not conduct structural biology research due to limited resources, lack of trained persons, and an exodus of skilled scientists. The most urgent requirement is to build on the emerging centres in Africa \u2013 some well-established, others growing. This can be achieved through workshops that improve networking, grow skills, and develop mechanisms for access to light source beamlines for defining X-ray structures across the continent. These would encourage the growth of structural biology, which is central to understanding biological functions and developing new antimicrobials and other drugs. In this light, a hands-on training workshop in structural biology series 4 was organised by BioStruct-Africa and the Malaria Research and Training Center (MRTC) in Bamako, Mali, to help bridge this gap. The workshop was hosted by MRTC from the 25th to 28th of April 2022. Through a series of lectures and practicals, the workshop enlightened the participants on how structural biology can be utilised to find solutions to the prevalent diseases in Africa. The short training gave them an overview of target selection, protein production and purification, structural determination techniques, and analysis in combination with high-throughput, structure-guided, fragment-based drug design. Summary: BioStruct-Africa has been building capacity in structural biology for Africa-based biologists and researchers. Visualizing biological molecules at the molecular level is essential in understanding how they perform their function . A strucBioStruct-Africa is a non-profit organization registered in Stockholm, Sweden (Swedish corporate ID: 802509-6689) whose mission is to build capacity for Africa-based researchers in the indispensable field of structural biology. The core team members are made up of, but not limited to, Africans with expertise in structural biology. BioStruct-Africa's inaugural workshop in 2019 was a saThe current BioStruct-Africa workshop was held at the Malaria Research and Training Center (MRTC) from the 25th to 28th of April 2022. The MRTC within the University of Science, Techniques and Technologies of Bamako is divided into several research groups and/or units, including the Genomics and Molecular Epidemiology, B-cell Laboratory, Cellular Immunology Laboratory, Molecular Epidemiology and Drug Resistance Unit, Clinical Drugs, and Vaccines Development units, Data Management and Analysis Group, Diagnostic Laboratories and Entomology Groups. During the past 30\u2005years, MRTC in collaboration with the US National Institutes of Health and various partners, has built state-of-the-art facilities for malaria and other pathogens research, including extensive \u221280\u00b0C freezer and liquid nitrogen storage facilities, parasite culture facilities, insectaries, genomic data storage and Bioinformatics facilities, etc.The BioStruct-Africa workshop hosted by MRTC was a hybrid event consisting of both lectures and hands-on training. The workshop's main goal was to give the participants insights into the significance of structural biology to containing the prevalent diseases in Africa. Specifically, the aim was to enable the participants to produce protein crystals by the traditional vapour diffusion method and remotely connect to a synchrotron beamline at the European Synchrotron Radiation Facility (ESRF) in Grenoble, France and collect X-ray diffraction data from crystals mounted on the beam by a beamline scientist. In addition, the participants were taken through the process of cryo-electron microscopy data collection and structural determination at the ESRF.Africa is the only continent without a synchrotron light source , which hThe BioStruct-Africa workshops are structured to contain both theoretical lectures and hands-on training in structural biology.https://www.ssgcid.org/). This workshop registered a higher number of female participants than the previous one, which could be because of our advertising strategy encouraging females to apply and providing a few stipends to support their participation.The workshop kick-started with an introduction to structural biology by Dr Michel Fodje . Dr Fodje's lecture mainly focused on the history of structural biology and the introduction of structural biology techniques, such as macromolecular crystallography, cryo-electron microscopy (cryo-EM), nuclear magnetic resonance spectroscopy, and small-angle X-ray scattering. Dr Fodje's lecture was followed by an overview of structural biology at the ESRF by Dr Daouda A.K. Traore . The theoretical lecture session continued with two lectures back-to-back by Dr Emmanuel Nji . In the first lecture, Dr Nji presented the strategies for selecting a suitable target for structural biology and the principles underlying a protein crystallization experiment. In the second lecture, Dr Nji presented the significance of structural biology for Africa. He specifically highlighted what approaches had been utilised that might prove useful in containing the prevalent diseases in Africa using a structure-based drug and vaccine design approach. Of particular interest during Dr Nji's second lecture was the number of structures solved in the pursuit of therapeutics for the COVID-19 pandemic in such a short time , highlighting the role played by structural biology in containing COVID-19 . LikewisFinally, Mikael Andersson Sch\u00f6nn presented the tricks of the trade for protein purification for crystallography purposes. It is not typically expected of structural biology scientists to also be protein purification experts. As such, having a relevant source of expertise in the field might help save projects with issues that otherwise seem insurmountable. His participation and including the industry in our course would help bridge the gap regarding the accessibility of consumables for protein purification experiments. Additionally, having a direct contact point to relevant companies through these workshops may provide future opportunities for university spin-off companies regarding industrial know-how.The participants were trained on vapour diffusion crystallization, crystal imaging, harvesting, and cryocooling using lysozyme as the protein of choice A. After The workshop was attended by 25 participants, mainly early-career scientists, Master\u2019s and PhD students, mostly from Mali. The participants took a pre-test at the beginning of the workshop and, a post-test at the end to judge the success of our teaching strategy. Based on the pre- and post-test results and student presentations, it was evident that the participants benefited from the workshop. Likewise, participants were given an online survey to complete anonymously about the pros and cons of the workshop, and the results shown in The vision of BioStruct-Africa is to ensure that the African continent actively conducts cutting-edge structural biology research. Research in structural biology would help provide the molecular details of disease intervention points and aid small molecule and vaccine development. Building capacity in structural biology for Africa-based researchers would ensure that Africans can utilise structural biology to find solutions to the diseases affecting their families, neighbours, and continent. In order to build capacity in structural biology for Africa-based researchers, BioStruct-Africa recently organised a workshop in structural biology at the MRTC, from the 25th to the 28th of April 2022. Overall, the workshop was very successful as participants were able to grow protein crystals and remotely connect to the European Synchrotron Radiation Facility, Grenoble, France, to collect X-ray diffraction data . They al"} +{"text": "Atrial fibrillation (AF), the most common sustained cardiac arrhythmia, has a large impact on quality of life and is associated with increased risk of hospitalization, morbidity, and mortality. Over the past two decades advances regarding the clinical epidemiology and management of AF have been established. Moreover, sex differences in the prevalence, incidence, prediction, pathophysiology, and prognosis of AF have been identified. Nevertheless, AF remains to be a complex and heterogeneous disorder and a comprehensive sex- and gender-specific approach to predict new-onset AF is lacking. The exponential growth in various sources of big data such as electrocardiograms, electronic health records, and wearable devices, carries the potential to improve AF risk prediction. Leveraging these big data sources by artificial intelligence (AI)-enabled approaches, in particular in a sex- and gender-specific manner, could lead to substantial advancements in AF prediction and ultimately prevention. We highlight the current status, premise, and potential of big data to improve sex- and gender-specific prediction of new-onset AF. Atrial fibrillation (AF), the most common cardiac arrhythmia, markedly increases the risk of hospitalization, morbidity, and mortality \u20133. Over The past decade has witnessed an exponential growth in recorded data in the healthcare sector. The massive amount of recorded information, i.e. big data, has turned to a topic of special interest, because of its great potential. Leverage of big data, using artificial intelligence (AI)-enabled approaches, provides an opportunity to further improve prediction of AF , 13. UseOver the past decade, several AF risk prediction scores have been developed and validated using more traditional research methods \u201322. ThesA variety of studies have highlighted the potential predictive capacity of AI to assess the risk of new-onset AF from a 12-lead ECG with acceptable to excellent performance (area under the receiver operating characteristic curve ranging between 0.70\u20130.90) , 33. TheTwo studies utilized EHRs to develop AF prediction models , 42. EHR2DS2-VASc score of AI and exact definition of how all the different methods work is yet difficult . This soLarge, diverse, and multidimensional data sources carry the potential to improve AF prediction and management. Yet, various challenges remain before AI-enabled algorithms can be adopted and implemented. Enhancement of personalized and precision medicine in AF warrants taking into account the complexity of sex and gender dimensions in big data sources and methods, while also overcoming the challenges that currently accompany the use of AI-enabled algorithms.Conceptualization, project administration, and validation: SG and MK. Data curation, investigation, methodology, resources, writing\u2014original draft, and writing\u2014review and editing: SG, ZL, and MK. Funding acquisition and supervision: MK. Software and visualization: SG. All authors contributed to the article and approved the submitted version.This study was further supported by the Senior Scientist Grant from Dutch Heart Foundation (03-004-2021-T050).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The Annual Conference and International Conference of the Chinese Association of Micro-NanoTechnology is a comprehensive, cross-disciplinary, high-level academic conference that has been held annually since 1994 and has become an important academic event in the field of micro- and nanotechnology. The conference includes an opening ceremony, reports related to the main and sub-venues, education and training, a call for papers, and technical exhibits. Moreover, it provides a platform for domestic and foreign micro- and nanotechnology workers to exchange ideas, technology, and scientific research pertaining to micro- and nanotechnology and related fields.This Special Issue contains sixteen papers and one review from the 23rd Annual Conference and 12th International Conference of the Chinese Society of Micro-Nano Technology (CSMNT 2021), which was held in Harbin, China (24\u201327 September 2021). The papers highlight new findings and technologies related to micro/nanoenergy, micro-electromechanical systems, nanosystems, and nanomaterials, as well as emerging related fields.This issue includes a proposal for a novel spiral-wound optic-fibre sensor to monitor the corrosion of steel bars by some authors; the feasibility of the spiral distributed sensors was verified experimentally. This method can be used to evaluate the initial and final cracking behaviours of concrete structures, as well as steel bar corrosion . Due to Light-emitting diodes (LEDs) are widely used in medicine, navigation, and landscape lighting. The development of high-power LEDs requires the dissipation of LED heat. The authors reviewed the packaging technology and structure in terms of the thermal performance of LED packaging and introduced related technologies that promote heat dissipation in LED packaging . Some auAnother group of authors used microelectromechanical systems (MEMS) combining electrical, optical, and mechanical components at the micro-scale for miniaturizing mechanical micro devices . The autAnother group of authors designed and fabricated a laser-controlled intelligent initiation system with inherent safety and a laser-controlled explosion-initiating device (LCEID) using integrated safe-and-arm, electromagnetic pulse-resistant, and fast-acting technologies. A modular design and integrated circuit fabrication techniques were also used . Some auWe would like to thank all of the authors for contributing their original manuscripts to this Special Issue, and the reviewers for their participation in the peer-review process, which helped to improve the quality of the papers."} +{"text": "The extension of life expectancy highlights the importance of understanding how people conceptualize \u2013 and plan for \u2013 their later years. We address this issue using data from an online survey of over 3,400 Floridians aged 50 and older that was conducted between December 2020 and March 2021 and funded by the Florida Department of Transportation. We examine five types of planning: for health care needs, financial well-being, living arrangements, driving retirement, and end-of-life care. We find that the likelihood of planning varies considerably across these types. Only 23 percent of respondents reported planning \u201csome\u201d or \u201ca lot\u201d for driving retirement, compared with 74 percent for health care needs, 76 percent for end-of-life care, 77 percent for living arrangements, and 83 percent for financial well-being. Likelihood of planning varied by age, gender, socioeconomic status, health, and race or ethnicity. Across all types of planning, older adults and those with at least a college degree and higher income were more likely to have planned. Women were more likely than men to plan for their financial future, living arrangements, driving retirement, and end-of-life care. Those in better health were more likely to plan for their financial future and end-of-life care. The effects of race or ethnicity were less consistent across the types of planning. White respondents were more likely than other race or ethnic groups to report planning for their living arrangements and end-of-life care, while Hispanic respondents were more likely than other groups to plan for driving retirement."} +{"text": "Sixteen people with PD and dysarthria completed LSVT LOUD followed by PD Check-Ins up until 24 months post-treatment. Self-rated QoL and voice handicap scales were used to determine the psychosocial and perceived impact of PD Check-In on the speech and voice of people with PD. The perceived impact of PD Check-In on speech and voice was also sought from 15 communication partners (CPs). A significant treatment effect for time was identified for the Dysarthria Impact Profile (DIP), Voice Handicap Index (VHI), and Voice Handicap Index-Partner (VHI-P) (p < 0.05). There was no significant effect for time for the Parkinson\u2019s Disease Questionnaire (PDQ-39). Planned comparisons of timepoints for DIP, VHI, and VHI-P showed no significant differences (p > 0.01). Comparison of perceived voice handicap by people with PD and CPs revealed no significant differences (p > 0.01). The impact of PD Check-In on QoL of people with PD and CPs for 24 months post-LSVT-LOUD is unclear. Self-reported outcome measures alone do not fully capture changes in QoL in PD.Quality of life (QoL) for people with Parkinson\u2019s Disease (PD) is diminished by speech and communication changes. The impact of PD Check-In, an intervention for supported self-managed maintenance of speech following LSVT LOUD Speech and communication changes associated with Parkinson\u2019s Disease (PD) contribute to a diminished quality of life (QoL) for those diagnosed with the progressive neurological condition, and their families and caregivers ,2. The ePeople with PD and their close communication partners (CPs) have described the considerable impact of deficits in speech and communication on their QoL. Changes in self-perception, such as feeling inadequate, less independent, self-conscious, less talkative, and unsure of relationships and dynamics of social interactions are experienced by people with PD in everyday communication ,12. EmotDespite the well-documented efficacy of the LSVT LOUD intensive speech treatment for PD ,19, longThe negative psychosocial impact of persistent speech and communication symptoms after SLP intervention, however, has been reported by people with PD in several studies. In a study by Spurgeon et al. , particiTo this end, PD Check-In, a novel intervention for SLP-supported self-managed maintenance of speech and communication for people with PD following LSVT LOUD, was developed and trialed in a clinical service for ambulatory rehabilitation. The development of PD Check-In and the underpinning framework has been described by Finnimore, Theodoros, and Rumbach . This moThis study aimed to investigate the impact of PD Check-In on the QoL for people with PD over twenty-four months following LSVT LOUD. Specifically, the study aimed to: (1) determine the self-reported psychosocial impact of speech and communication changes for people with PD; (2) determine the perceived impact of voice and speech changes on the QoL of people with PD from the perspective of CPs; and (3) compare the perceived impact of voice and speech changes of CPs and people with PD over time. It was hypothesized that a SLP-supported self-managed maintenance program following LSVT LOUD would improve QoL of people with PD. Additionally, it was hypothesized that the perceived impact of voice and speech changes for people with PD would align with those of their CPs.A small group repeated measures Phase 1 study design was selected for this preliminary investigation in a clinical setting .Recruitment of two cohorts of participants was conducted as a sample of convenience in an ambulatory rehabilitation setting. Twenty people with hypokinetic dysarthria due to PD, and 19 close CPs, met eligibility criteria for the study. All participants were 18 years or older, proficient in English, and independent in providing informed consent. All participants with PD had a confirmed diagnosis of idiopathic PD with dysarthria provided by a neurologist or medical practitioner with expertise in PD. People with a history of neurosurgical management of PD, including Deep Brain Stimulation (DBS), remained eligible to participate. Exclusion criteria were applied to potential participants with PD with severe cognitive impairment, including diagnosed dementia and co-existent severe medical, neurological, and psychiatric conditions. Further prerequisites for inclusion of people with PD related to suitability for LSVT LOUD, which was established prior to recruitment to the study in an initial SLP assessment conducted by a clinician independent of the research team. Additionally, confirmation of vocal symptoms consistent with PD, and suitability of people with PD for intensive voice treatment were obtained by an assessment performed by an otolaryngologist, independent of the study. From the PD cohort, there was 20% (n = 4) attrition of participants from the study, leaving a data set of 16 participants with PD. Reasons for withdrawal related to revision of neurological diagnosis (n = 2), seeking additional SLP services (n = 1), and relocation inter-state (n = 1). Nine men and seven women with a mean age of 70.68 years comprised the PD cohort. On average, PD participants were 5.9 years post-diagnosis . The median stage of PD progression was 2.5, with a range from Stage 1\u20134 . Two peoClose CPs, unfunded for their companionship, were nominated by participants with PD. CPs with communication impairment, including cognitive and hearing deficits, were not eligible for inclusion. The recruited cohort of CPs comprised 19 participants, as one participant with PD did not identify a close CP. The complete data set of 16 CPs remained after attrition of three participants secondary to the withdrawal of three participants with PD from the study, and the withdrawal of a fourth CP due to frailty see .Participants with PD received the LSVT LOUD program followed by PD Check-In, a maintenance intervention, delivered at 6 and 12 weeks, and 6, 12, and 24 months post-LSVT-LOUD. Each individual face-to-face PD Check-In session, delivered by the principal investigator, lasted approximately one hour. CPs were invited to attend PD Check-In sessions at the discretion of the participants with PD. LSVT LOUD was delivered in accordance with protocol by an accredited SLP . Pre- anA typical PD Check-In session comprised the clinical evaluation of vocal intensity of phonation and speech and fundamental frequency range of the person with PD, followed by a semi-structured discussion between the SLP and the person with PD, possibly accompanied by their CP, targeting the development of self-management principles for long-term speech maintenance. Clinical outcomes formed the basis of collaborative reflection on the maintenance of vocal intensity using comparative data from pre- and post-LSVT-LOUD. Clinical issues identified by the SLP regarding voice production received therapeutic advice and intervention. The semi-structured discussion, based on a topic guide , explorePaper-based standardized self-rating scales were provided to participants with PD and CPs, at varying timepoints from pre-LSVT-LOUD to 24 months post-treatment see , measuriThe DIP measures the psychosocial impact of dysarthria from the perspective of the speaker. The questionnaire comprises 48 questions presented in five sections: (A) the effect of dysarthria on me as a person; (B) accepting my dysarthria; (C) how I feel others react to my speech; (D) how dysarthria affects my communication with others; and (E) dysarthria relative to other worries and concerns . StatemePDQ-39 explores the impact of PD on QoL through a standardized self-rated questionnaire across eight health and lifestyle domains. People with PD are required to respond based on their experiences in the period one month prior to completing the questionnaire. The domains are presented with a variable number of: mobility (10), activities of daily living (6), emotional wellbeing (6), stigma (4), social support (3), cognitive impairment (4), communication (3), and bodily discomfort (3). Participants were required to independently rate their response with a mark for each statement using a 5-point ordinal scale . The total score for each domain was calculated by dividing the sum of the participant scores by the potential score for the domain multiplied by 100. A Summary Index Score was calculated as the sum of the total scores in each domain. A lower score on PDQ-39 represents a better QoL for the person with PD . For theThe VHI is a questionnaire seeking the speaker\u2019s perception of the handicap they experience due to their disordered voice . The 30-CPs provided their perceptions of the handicap experienced by people with PD by completing the VHI-P at 12 anp < 0.01 was used to account for multiplicity of testing [p < 0.01 to account for multiplicity of testing to determine if a significant difference in perceived voice and speech handicap for people with PD existed between participants with PD and CPs across time points.Data analyses were performed using SPSS . Non-parametric procedures were performed for the ordinal data from each measure. Data from the DIP, PDQ-39, VHI, and VHI-P were analyzed using the Friedman Test with an alpha level of 0.05 to determine whether PD Check-In had a significant effect on QoL over time. Where significance was found, planned comparisons using Wilcoxon signed-rank tests were conducted at pre-to-post, post-to-12-m, and post-to-24-m. A stringent alpha level of testing . Data fr2 = 319.12; p = 0.0001), VHI , and VHI-P . No significant effect for time was identified for the PDQ-39 . Planned comparisons of the DIP, VHI, and VHI-P data using the Wilcoxon signed-rank test and an alpha level of p < 0.01 failed to identify any significant differences across the time points for these outcome measures of conversational monologue 24 months following LSVT LOUD . ChangesThough the DIP, PDQ-39, and VHI have been used extensively as validated patient reported outcomes in studies in PD, speech, and communication ,41, measThe DIP, PDQ-39, VHI, and VHI-P shared limitations in the extent to which they fully represented the impact of speech and communication changes on QoL for people with PD. The DIP extensively probed the impact of features of dysarthria on QoL without specificity for PD . The comThe limitations of PDQ-39 for this study were largely related to the breadth of the scale . In ordeFor people with PD, self-perception of the clinical features of articulation, and voice quality may be less developed than their awareness of overall changes in communication, and loss of speech clarity in functional contexts ,16. AlthThe perspective of CPs on the impact of voice and speech changes on the QoL of people with PD over time was investigated. Though changes in CPs\u2019 perception across time points on VHI-P were not significant, there was a trend of lessened perceived impact of speech in all domains from pre- to post-LSVT-LOUD. Importantly, at 24 months post-treatment, the perceived lessened impact of speech on quality of life for participants with PD remained above the pre-treatment level in all domains. This trend in CPs\u2019 perception is suggestive of the importance of the maintenance of voice and speech following treatment for QoL for people with PD and CPs.A further aim of this study was to compare the perceptions of CPs with those of people with PD regarding the impact of voice and speech changes on QoL for people with PD over time. Previous studies which compared self-reported perceptions of voice handicap reported CPs generally perceived less impact of speech changes on the QoL of people with PD; however, no statistically significant differences were found ,17. ThouThe influence of PD Check-In on the perceived impact of speech on the QoL is not clear; however, the descriptive data presented in Elements of the design and conduct of the study may have limited the capturing of the QoL impact of PD Check-In for people with PD and CPs. As a Phase 1 study , a smallThe paper-based questionnaires were completed by participants in the latter part of a PD Check-In session, following the clinical outcome measures and semi-structured discussion. The number of questionnaires presented varied according to the time point post-LSVT-LOUD see ; howeverThe design of the questionnaires and the paper-based format proved to be functionally challenging for some participants due to writing and visual difficulties associated with PD ,45 resulLimitations of individual measures have been discussed. In the time since this longitudinal study commenced, considerable advances have been made in the validation of tools which enable SLPs to more effectively assess the psychosocial influences on speech and communication, and to evaluate the impact of intervention on QoL ,41,45. TThe impact of PD Check-In on the QoL of people with PD and their CPs over 24 months following LSVT LOUD was inconclusive. Though sustained, if not measurably improved, QoL through supported maintenance of speech and communication following treatment is the primary goal for this intervention, the evaluation of QoL is compounded by the individuality of the experience of PD, and the unique factors which determine QoL for individuals. The development of measures which capture the individual communication and social participation experiences of people with PD, and accommodate the physical, emotional, and cognitive barriers faced by them and their CPs, is necessary for engagement with self-reported QoL measures. PD Check-In offers long term SLP connection for people with PD and CPs following treatment, employing a balance of clinical and psychosocial communication goals for a better life with PD. Qualitative analysis of the semi-structured discussion component of PD Check-In may enhance our understanding of QoL for people with PD and their CPs, and contribute to a refinement of evaluation measures which encompass the impact of speech and communication intervention in PD. Future studies in the form of randomized controlled trials are required to further investigate the efficacy of PD Check-in as a model for speech maintenance and QoL for people with PD and their families and CPs."} +{"text": "The epidemiologic and medical communities have witnessed a parallel story of HIV/AIDS and Kaposi Sarcoma (KS) unfold for the better part of 4 decades. Prior to the 1980s, KS was rare in the United States and the In the 1990s, KS incidence sharply declined, which was attributed to a decrease in HIV/AIDS-related immunosuppression due to both the introduction of antiretroviral therapy and how JNCICS, Suk et al. .Role of the funder: The funder had no role in this editorial.Disclosures: The authors have no disclosures.Author contributions: Conceptualization, writing\u2014original draft, writing\u2014review and editing: ASR, KSP."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "No comprehensive studies have been published on the global burden of alopecia areata since 2010.We aimed to measure the global, regional, and national incidence of alopecia areata and disability-adjusted life-years (DALYs) by age, sex, and socio-demographic index (SDI) value from 1990 to 2019.Data were extracted from the Global Burden of Disease Study 2019. Estimated annual percentage changes (EAPCs) were calculated to quantify temporal trends in the age-standardized rates of alopecia areata incidence and DALYs. The correlations between EAPCs in the age-standardized rates and SDI values were also analyzed.From 1990 to 2019, the alopecia areata incidence number and the associated number of DALYs increased globally by 49.14%, and 49.51%, respectively. The global age-standardized incidence rate decreased and the age-standardized DALY rate showed a downward trend . The largest increases in the age-standardized incidence rate and age-standardized DALY rate were observed in Low SDI quintile and Western Sub-Saharan Africa regions. The regions with the greatest changes in the incidence of alopecia areata were Central Sub-Saharan Africa and Western Sub-Saharan Africa. The three countries with the largest increases in alopecia areata incidence from 1990 to 2019 were Kuwait , South Sudan , and Nigeria . The age-standardized incidence rate was higher in females than in males.Globally, both the age-standardized incidence rate and age-standardized DALY rate of alopecia areata showed decreasing trends. Future preventive strategies should focus on low-income countries, Central Sub-Saharan Africa, Western Sub-Saharan Africa, Kuwait, South Sudan, Nigeria. Alopecia areata is a common chronic tissue-specific autoimmune disease that causes patchy hair loss. It affects 2% of the general population . In someRecent studies have only presented the burden of alopecia areata based on regional and/or national data or data from the Global Burden of Disease (GBD) 2010 study , and havIn this study, to provide comprehensive and comparable information on the burden of alopecia areata, we analyzed the global, regional, and national incidence and disability-adjusted life-years (DALYs) data from the GBD 2019 study in terms of counts and age-standardized rates by sex, age, and socio-demographic index (SDI) value.http://ghdx.healthdata.org/gbd-results-tool). The data were from 204 countries and territories stratified by age and sex from 1990 to 2019 . The number of years lived with disability was calculated as the product of the disability weight and the prevalence of alopecia areata. The number of DALYs due to alopecia areata were calculated as the sum of the number of years lived with disability and the years of life lost due to premature death. Final estimates were computed using the mean estimates across 1,000 draws, and 95% uncertainty intervals (UIs) were specified on the basis of the 25Estimated annual percentage changes (EAPCs) were calculated to quantify trends in the incidence of alopecia areata and the number of DALYs. The natural logarithm of the regression line fitted to the age-standardized rate was y = a + bx + c, where x is the calendar year . The EAPGlobally, the incidence number of alopecia areata increased from 21742836.45 in 1990 to 32426829.18 in 2019, whereas the related number of DALYs increased from 401682.16 in 1990 to 600570.37 in 2019. Based on these values, the incidence of alopecia areata and the number of DALYs increased by 49.14%, and 49.51%, respectively, from 1990 to 2019.From 1990 to 2019, the age-standardized incidence rate of alopecia areata and the age-standardized DALY rate showed a downward trend globally and Western Sub-Saharan Africa (1.47), whereas the region with the smallest change was Central Europe \u22120.04, Table\u00a01.The male-to-female ratio of alopecia areata incidence peaked in the 20\u201324-year age group globally and in high-SDI, high-middle-SDI, middle-SDI, and low-middle-SDI regions, but in the 90\u201394-year age group in low-SDI regions , South Sudan , and Nigeria , Southern Latin America (9.41), and High-income Asia Pacific .The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.5Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.13The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.14Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.18The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Medical image segmentation is an essential component of computer-aided diagnosis (CAD) systems. Thyroid nodule segmentation using ultrasound images is a necessary step for the early diagnosis of thyroid diseases. An encoder-decoder based deep convolutional neural network (DCNN), like U-Net architecture and its variants, has been extensively used to deal with medical image segmentation tasks. In this article, we propose a novel N-shape dense fully convolutional neural network for medical image segmentation, referred to as N-Net. The proposed framework is composed of three major components: a multi-scale input layer, an attention guidance module, and an innovative stackable dilated convolution (SDC) block. First, we apply the multi-scale input layer to construct an image pyramid, which achieves multi-level receiver field sizes and obtains rich feature representation. After that, the U-shape convolutional network is employed as the backbone structure. Moreover, we use the attention guidance module to filter the features before several skip connections, which can transfer structural information from previous feature maps to the following layers. This module can also remove noise and reduce the negative impact of the background. Finally, we propose a stackable dilated convolution (SDC) block, which is able to capture deep semantic features that may be lost in bilinear upsampling. We have evaluated the proposed N-Net framework on a thyroid nodule ultrasound image dataset and the DDTI publicly available dataset. The experimental results show that our N-Net model outperforms several state-of-the-art methods in the thyroid nodule segmentation tasks. The thyroid gland is a butterfly-shaped endocrine gland, which lies in the anterior part of the neck just below the thyroid cartilage are the most widespread approaches for the diagnosis of thyroid diseases. In clinical practice, ultrasound examinations are dependent on visual inspection by experienced clinicians, which requires concentrated attention and a high level of skill. The diagnosis process is time-consuming, labor-intensive, and prone to operation bias. Furthermore, it is hard to identify the subtle differences between malignant and benign nodules. Geometry and margins of thyroid nodules are the key features to distinguishing between malignant and benign nodules has been widely used in the field of medical image segmentation block to solve the problem of restricted receptive fields. Meanwhile, we added the attention guidance (AG) module before the jump connection structure to establish long-range dependencies, and introduce a multi-scale mechanism to solve the problem of ignoring the global contextual information of different scales. In this study, we propose a combined fully convolutional network for thyroid nodule segmentation using ultrasound images, referred to as N-Net. The main contributions of this study are 3-fold as follows:1) We propose an N-shape fully convolutional network, which contains a multi-scale U-shape convolutional network to learn multi-scale feature representations. The multi-scale input layer is used to construct an image pyramid input, which can achieve multi-level receiver field sizes, thus learning rich feature representations. This makes the network learn both the global and the local information of thyroid ultrasound images to extract the multi-level features.2) An attention guidance module is proposed to preserve the structural semantic features to improve the segmentation performance. This module makes the model pay more attention to the thyroid regions of the input image and reduces the effect of mixed noise in ultrasound images on our network. It can be considered as semantic guidance in the network to acquire more precise semantic representations.3) We designed a stackable dilated convolution (SDC) block to encode the high-level semantic contextual features from feature maps. The SDC block adopts a hybrid dilated convolution with different dilation rates. Our network can learn the semantic features which may be lost in bilinear upsampling through the SDC block.The remainder of this article is organized as follows. Section 2 introduces related study. Section 3 describes the proposed method in detail. Section 4 presents the experiment results. Conclusions are summarized in Section 5.Traditional segmentation methods include contour and shape based methods and region based methods. Tsantis et al. used the morphological and wavelet-based features of nodules in thyroid ultrasound images to assess the malignancy risk of thyroid nodules on ultrasonography in 2009 was originally proposed for the efficient computation of the wavelet transform block to encode the high-level semantic contextual features from feature maps. Compared with the standard convolutional layer, the dilated convolution uses a larger convolutional kernel and enlarges the receptive field without reducing the resolution of the feature map. In the semantic segmentation network, the larger the receptive field of the final predicted pixels, the better segmentation performance the deep network can achieve. In addition, it should be mentioned that the use of dilated convolution does not increase the number of parameters and computations.via a skip connection. In general, the convolution of a large reception field can extract and generate abstract features for large objects, while the convolution of a small reception field is good at segmenting small objects. By stacking the dilated convolution with different dilation rates, the SDC block is able to allow the network to learn contextual information and extract features for objects of various sizes. The four cascaded dilated convolution branches are added to the feature map itself and then sent to the subsequent decoding module. The input size is the same as the output size in the SDC module.As shown in The TNUI-2021 dataset was used to evaluate our N-Net, which consists of 1,381 ultrasound thyroid nodule images. The resolution of each image is 780 \u00d7 780. This dataset was acquired from 483 patients by doctors in the Fujian Medical University Union Hospital using two apparatuses, Supersonic Aixplorer and SAMSUNG WS80A. In the TNUI-2021 dataset, each image is paired with ground truth (GT) image. The GT images were manually labeled by the expert pathologists in the Fujian Medical University Union Hospital. The labels were stored in tabular files, and benign and malignant classifications were also labeled. There are 72 images of benign nodules and 1,309 images of malignant nodules.The DDTI dataset contains 637 ultrasound thyroid nodule images of different resolution sizes, such as 560 \u00d7 360, 280 \u00d7 360, and 245 \u00d7 360. The ratio of the training set, validation set, and test set is 6:2:2, and the augmented patches are resized to the size of 512 \u00d7 512 for training.Considering the serious imbalance of these two types of data, we aggregated the data of benign and malignant nodules to divide the data set. We employed a 5-fold cross-validation method to evaluate the performance of our N-net. Specifically, the TNUI-2021 dataset was randomly and equally divided into five non-overlapping sub-datasets. For each time, 20% of images were used for testing, and the remaining 80% of images were used for training and validation. The ratio of the training set, validation set, and test set in each experiment was 6:2:2. The averaged results of five validations were obtained as the final results. The N-Net training and testing were completed in PyTorch. The training and testing platform was the Ubuntu 18.04 system with an NVIDIA GeForce RTX 2070 graphics card, which has 8 Gigabyte memory.In our N-Net, in order to reduce the risk of overfitting, we enlarged the training dataset with the online data augmentation processing, including rotation of images at 90, 180, and 270-degree angles, horizontal flip, and vertical flip. Then we resized the augmented patches to the size of 512 \u00d7 512 for training. The Adam optimizer , mean intersection over union (mIoU), Precision, Recall, and F1-Score, which were calculated as follows:where TP, FP, and FN denote the number of true positives, false positives, and false negatives, respectively. We used the Dice, mIoU, Precision, Recall, and F1-Score to measure the segmentation performance.We compared the proposed N-Net model with several different segmentation approaches in In addition, in terms of visualized segmentations as shown in Ablation studies were performed to analyze the effectiveness and the contributions of each module in the proposed N-Net model on the TNUI-2021 dataset. In addition, in terms of visualized segmentations, the results are shown in Our proposed N-Net model employs the U-Net block, which is able to capture and encode deeper semantic features that may be lost in bilinear upsampling. Experimental results on the TNUI-2021 dataset and the DDTI dataset demonstrate that the proposed N-Net model outperforms several typical segmentation approaches.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Ethics Committee of Fujian Medical University Union Hospital. The patients/participants provided their written informed consent to participate in this study. Written informed consent was obtained from the individual(s) for the publication of any potentially identifiable images or data included in this article.XN, XZ, TT, XL, LW, HZ, JL, SC, and MD: concept and design. XN, XZ, HZ, EX, SC, MZ, CC, and TT: acquisition of data. XN, XZ, TT, XL, LW, and SC: model design and manuscript drafting. XN, XZ, XL, LW, and TT: data analysis. XN, XZ, TT, XL, LW, HZ, JL, EX, SC, MZ, CC, and MD: approval. All authors contributed to the article and approved the submitted version.This study was supported by the National Natural Science Foundation of China under (Grant Nos. 61901120 and 62171133), sponsored by Fujian Provincial Health Technology Project (2019-1-33), in part by the Science and Technology Innovation Joint Fund Program of Fujian Province of China under (Grant No. 2019Y9104), and the Guiding Projects of Fujian Provincial Technology Research and Development Program under (Grant No. 2022Y0023).Author TT was employed by Imperial Vision Technology. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Tuberculosis (TB) always runs in the forefront of the global burden when it comes to infectious diseases. Tuberculosis, which can lead to impairment of quality of life, financial hardship, discrimination, marginalization, and social barriers, is a major public health problem. The assessment of TB burden and trend can provide crucial information for policy decision and planning, and help countries in the world to achieve the goal of sustainable development of ending the epidemic of TB in 2030.All data are from the Global Burden of Disease 2019 (GBD 2019) database, which analyzed the burden trend of age-standardized incidence, DALYs, and deaths rate in TB and HIV/AIDS-infected TB over the past 30 years. Also, GBD 2019 not only analyzed the burden distribution of TB in 204 countries and main regions of the world but also analyzed the relationship between the burden of global TB and the socio\u2013demographic Index (SDI).The age-standardized incidence, age-standardized disability-adjusted life years (DALYs), and age-standardized deaths rate for HIV-negative TB were 10,671.45 , 59,042.45 , and 1,463.62 in 2019, respectively. Age-standardized incidence, age-standardized DALYs, and age-standardized deaths rate of HIV/AIDS-XDR-TB were 2.10 (1.51\u20132.90), 64.23 (28.64\u2013117.74), and 1.01 (0.42\u20131.86), respectively. We found that TB is inversely proportional to SDI, the age-standardized incidence, DALYs, and deaths rate low burden countries were in high SDI areas, while high burden countries were in low SDI areas. The global TB showed a slow decline trend, but the age-standardized incidence of HIV-positive TB was increasing, and mainly distributed in sub-Saharan Africa.Age-standardized incidence, age-standardized DALYs, and age-standardized deaths rate of TB is related to SDI, and the burden of low SDI countries is lighter than that of high SDI countries. Without effective measures, it will be difficult for countries around the world to achieve the goal of ending the TB epidemic by 2030. Effective control of the spread of TB requires concerted efforts from all countries in the world, especially in the countries with low SDI, which need to improve the diagnosis and preventive measures of TB and improve the control of HIV/AIDS-TB. Tuberculosis is an ancient disease. In 1882, Robert Koch announced the discovery of the main bacterium of TB and named it Mycobacterium TB . TubercuAlthough TB has dropped from No. 7 position in 1990 to the No. 20 position in 2019 , percent. If the This study can provide the latest data on the age-standardized incidence, DALYs, and mortality of global TB through GBD 2019 data, which is of great significance for the prevention and control of tuberculosis. Combined with age-standardized incidence rates of TB, the association of DALYs and deaths rate analyses with SDI was more reflective of the impact of TB on socioeconomic status. Moreover, the GBD 2019 database can analyze the changes of TB in 204 countries and regions in the past 30 years (1990\u20132019). The burden of TB is related to many factors, such as gender, location, AIDS, and drug resistance. Analyzing the correlation between TB and these factors through the GBD 2019 database system and updating relevant information in a timely manner will help control the burden of TB and epidemiological trends.http://ghdx.healthdata.org/gbd-2019). The GBD 2019 includes 369 diseases and injuries in 204 countries or regions around the world as well as more than 80 behavioral, environmental, and other risk factors, whose estimation of attributable burden followed the general framework established for the comparative risk assessment (CRA) , 12 usedThe disability-adjusted life years (DALYs) was proposed by Global Burden of Disease to measure the disease burden, whose calculation takes into account the sum of years of life lost (YLL) and years of disability (YLD) for each reason, age, location, duration, gender, and year , 13. Thehttp://ghdx.healthdata.org/gbd-2019) is a comprehensive indicator that reflects the social conditions and population, including per capita income, average years of education, and total fertility rate. The SDI score ranges from 0 to 1, which means that the correspondence relationship from the lowest income, the lowest average years of education, and the highest fertility rate to the highest income, the highest average years of education, and the lowest fertility rate, and each location is assigned an SDI score every year. The SDI was developed for GBD 2015 was statistically significant. When reporting uncertainty intervals, the 95% UIs were calculated using the 2.5th and 97.5th percentiles of the draw-level values.We adopted the same techniques used elsewhere in the GBD study design to propagate uncertainty \u201321. For From 1990 to 2019, according to the GBD 2019 age-standardized incidence, DALYs, and deaths rate estimates, all HIV-negative TB including the drug-susceptible tuberculosis (DS-TB), multidrug-resistant tuberculosis without extensive drug resistance (MDR-TB without XDR), and extensively drug-resistant tuberculosis (XDR-TB) were analyzed. The results of age-standardized incidence, DALYs, and deaths rate all show that DS-TB ranks first among the three types of HIV-negative TB, followed by MDR-TB without XDR and XDR-TB . For HIVFrom 1990\u20132019, the change of age-standardized incidence, DALYs, and deaths rate of all-HIV negative TB have allAccording to GBD 2019, we analyzed the age-standardized incidence of all HIV negative TB in region-specific and country-specific in 2019 . Among mFrom 1990 to 2019, the change of age-standardized incidence rate of all HIV-negative TB in 204 cIn 2017, the incidence (in thousands) and prevalence (in thousands) of DS-TB were 8,508.6 and 9,828.6 , respectively . In 2015In 2019, age-standardized incidence, DALYs, and deaths rate of DS-TB 95% CI, are 10.0In 2019, in high SDI and low SDI regions, the age-standardized incidence rate of DS-TB ], followed by Sub-Saharan Africa and Eastern Sub-Saharan Africa (both are >200 million) . For ageIn 2019, the age-standardized incidence rate of DS-TB is different in different countries and regions ] , EasternIn 2019, the age-standardized incidence rate of MDR-TB without XDR and in the WHO2020 report . The proOur research still has some limitations. (1) The data in GBD 2019 are estimates, and the lack of overall HIV-positive TB data is not conducive to the assessment of overall HIV-positive TB. (2) In 1990\u20132010, the data of change of HIV/AIDS-MDR-TB without XDR incidence, DALYs and deaths rate is insufficient in some countries and regions. (3) This study did not analyze the epidemiology of TB, the incidence of TB in various age groups, DALYs and deaths rates, including the status of potentially infected TB. Nevertheless, GBD2019 still shows us a wealth of data for researchers to use, macroscopically showing the trend and burden of TB. We conduct a comprehensive and systematic assessment of the global burden of TB, which provides vital information for reducing the burden of TB.In 2017, low-income and middle-income countries spent US$10.90 billion (10.3\u201311.8) on TB and US$20.20 billion (17.0\u201325.0) on HIV/AIDS . World HThe original contributions presented in the study are included in the article/C-YJ, JiaZ, Z-hF, and YX: study conception and design. YX, JiaZ, PW, and J-hL: data acquisition and analysis. JieZ and B-NX: organize the article table section. W-qL and Y-YF: draw the picture. C-YJ: study supervision. Z-hF: administrative support. YX: drafting the manuscripts. C-YJ, JiaZ, and Z-hF: critical revision of the manuscript. All authors contributed to the article and approved the submitted version.This study was supported by the Starting Package of Xiang'an Hospital of Xiamen University (PM201809170010), Open project of Provincial Key Laboratory of Union Hospital Affiliated to Fujian Medical University in 2020 (Nos. XHZDSYS202004 and XHZDSYS202005), and Xiamen municipal Bureau of Science and Technology Grant (3502Z20174079), and the National Natural Science Foundation of China (Grant numbers: 82003178).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Natural toxins include a wide range of toxic metabolites also occurring in food and products, thus representing a risk for consumer health. In the last few decades, several robust and sensitive analytical methods able to determine their occurrence in food have been developed. Liquid chromatography mass spectrometry is the most powerful tool for the simultaneous detection of these toxins due to its advantages in terms of sensitivity and selectivity. A comprehensive review on the most relevant papers on methods based on liquid chromatography mass spectrometry for the analysis of mycotoxins, alkaloids, marine toxins, glycoalkaloids, cyanogenic glycosides and furocoumarins in food is reported herein. Specifically, a literature search from 2011 to 2021 was carried out, selecting a total of 96 papers. Different approaches to sample preparation, chromatographic separation and detection mode are discussed. Particular attention is given to the analytical performance characteristics obtained in the validation process and the relevant application to real samples. Natural toxins include a wide range of toxic metabolites which are synthesized by various organisms such as animals, certain plant species or by microorganisms. They may endogenously occur when produced by organisms commonly present in food or exogenously occur when produced during the metabolism of living organisms ,2,3. In The reduction of risks related to the presence of natural toxins in food plays an essential role in protecting consumers. Indeed, WHO, together with the European Food Safety Authority (EFSA), FAO and Codex Alimentarius Commission, have established maximum residue limits (MRLs) or recommendations for many of these natural toxins to control their occurrence in food ,12,13,14Different sample preparation strategies, including extraction, purification (clean-up) and preconcentration procedures have been proposed in the literature to eliminate possible interferences and enrich the sample. Solid phase extraction (SPE) and liquid\u2013liquid extraction (LLE) have been more frequently applied for clean-up procedures. Among SPE, the most currently used extractive phases are those based on C18, polymeric and antibodies anchored onto a support material sorbent. Alternatively, the simple dilution of the sample (dilute and shoot) or protein precipitation have been used in the case of acceptable sensitivity of the method .For the separation and final determination of natural toxins, methods based on high performance liquid chromatography (HPLC) coupled to fluorescence or ultraviolet (UV) detection have been widely used for the analysis of single or small groups of structural related toxins in food products . HPLC-baThe use of LC-based methods, which can lead to the coelution of some analytes, without providing any structural information, and the need to detect a great multiplicity of analytes in a single run, have shifted this field towards the use of LC in combination with mass spectrometry (LC-MS). Nowadays, LC-MS is the most powerful technique for the simultaneous detection of multiple regulated, unregulated, and emerging toxins in one single run due to its excellent sensitivity even at low concentration levels, selectivity, and its ability to resolve co-eluting compounds based on their molecular masses . Most ofThe chromatographic separation of toxins is commonly carried out through reversed-phase columns, even though polar and ionizable analytes can better be retained/separated by other elution modes, such as hydrophilic interactions chromatography (HILIC). The LC-MS methods for the quantitative determination of natural toxins are commonly based on the use of triple-quadrupole analyzer, tandem mass spectrometry, with multiple reaction monitoring (MRM) mode, allowing to simultaneously analyze several compounds with high sensitivity, selectivity and accuracy . To fulfThe co-occurrence of natural toxins in combination with other chemical contaminants such as pesticides, growth regulators and veterinary drugs and bioactive compounds , in a broad range of food matrices has driven an increasing interest for analytical methods addressing the simultaneous determination of multiple analyte classes. The simultaneous determination of multiclass analytes in complex food matrices is commonly based on LC-MS analysis, either using triple quadrupole or high-resolution approaches, thanks to their advantages in terms of selectivity and sensitivity .www.scopus.com, accessed on 31 January 2022) and keywords used for the search were \u201cliquid chromatography mass spectrometry\u201d and \u201cfood\u201d; then, the keywords \u201cmycotoxins\u201c, \u201calkaloids\u201d, \u201cmarine toxins\u201d, \u201cglycoalkaloids\u201d, \u201ccyanogenic glycosides\u201d and \u201cfurocoumarins\u201d were used to search the relevant literature for these particular groups of natural toxins in the selected years to identify all publications using LC-MS based methods. In this way, we found numerous articles, book chapters, and seminar proceedings. A total of 1966 papers were published in the selected period with a positive trend over the years and with mycotoxins representing the most abundant class (n = 1122), followed by alkaloids (n = 459), marine toxins (n = 244), and to a lesser extent glycoalkaloids (n = 22), furocoumarins (n = 10) and cyanogenic glycosides (n = 9) peptides were not included in this review. The literature search employed the Scopus online database ( (n = 9) .For all the classes, and in particular for the most abundant ones, a further refinement of selected papers was applied in order to select only those describing the development and validation of an LC-MS method . The novelty of the used technology, the application to new food matrices and the possibility to simultaneously analyze multiple toxins also including emerging ones were taken into account for the selection. In the case of mycotoxins, due to the high number of publications in the period 2011\u20132021, the literature search was refined to a more restricted period . A total of 96 papers were considered for this review by subdividing them in six specific sections, and in a further one for application to multiclass analysis.Aspergillus, Penicillium, Fusarium, Claviceps and Alternaria species. These toxins can contaminate various agricultural commodities either before harvest or under postharvest conditions, thus posing a risk to human and animal health due to their toxic effects. In addition, significant losses of yields and quality of the infested commodity are observed. Among the over 300 mycotoxins that have been identified, those of major concern worldwide causing food-borne illnesses are aflatoxins, ochratoxin A, fumonisins, zearalenone, patulin, citrinin, type B trichothecenes, mainly deoxynivalenol and nivalenol, and type A trichothecenes, mainly T-2 and HT-2 toxins. Their toxic effects range from nephrotoxicity, cytotoxicity, nervous-system disturbances, gastrointestinal diseases, to immunotoxicity, mutagenicity, teratogenicity and carcinogenicity [Mycotoxins are a group of natural compounds produced under a wide range of climatic conditions by filamentous fungi mainly belonging to genicity ,20. In pgenicity ,24,25,26Alternaria toxins [After the infection of crop plants, mycotoxins are modified by plant enzymes and are often conjugated to more polar substances. These substances are usually not detected during routine analysis, are unregulated, and are called \u201cbound\u201d or alternatively \u201chidden\u201d, \u201cconjugated\u201d or \u201cmasked\u201d and more recently \u201cmodified\u201d mycotoxins. Some metabolites are more toxic than the parent compound, while others are less toxic. Furthermore, depending on the type of linkage with matrix component, a part of bound mycotoxins could become bioavailable again in the digestive tract of humans and animals, thus contributing to the toxicity of parent compound . Other ua toxins ,30,31,32a toxins .Several chromatographic methods, mainly based on HPLC coupled with UV/diode array and fluorescence detection, have been developed and extensively reviewed for the determination of single mycotoxin or structurally related mycotoxins in food and feed. Gas chromatography (GC) and gas chromatography coupled to mass spectrometry (GC-MS) are also available for the specific detection of trichothecenes ,35,36. n) and high-resolution MS (HRMS) can provide structural and accurate mass information not only for the determination of well-known mycotoxins with remarkable sensitivity and specificity, but also for the analysis of emerging and modified mycotoxins [The evidence of co-occurring mycotoxins in various matrices has led to the development of new multi-mycotoxin methods for their simultaneous detection in the same matrix. The absence of structural information, as well as the coelution of structurally related mycotoxins, have shifted this field towards more sophisticated detection techniques based on the use of MS detectors. As a consequence, MS coupled with HPLC and ultra-HPLC systems has turned into one of the most powerful tools for multi-mycotoxin analysis at very low concentrations in complex matrices ,37. Furtcotoxins ,36,38. Ahttps://standards.cencenelec.eu/ accessed on 5 April 2022). The complexity of the matrices, mainly including those of animal origin, has led to the development and validation of specific analytical protocols allowing for quantitative extraction and detection of targeted mycotoxins. The availability of standardized methods of analysis is of paramount importance to guarantee a uniform application of the EU legislation and contribute to maintaining a high level of food and feed safety. Despite the huge number of published multimycotoxin LC-MS methods, their implementation in control laboratories has been limited in the past years due to their performance characteristics not fulfilling the acceptability criteria for mycotoxins established at the EU level . In 2013A multi-mycotoxin analysis method based on UPLC-ESI-MS/MS for the determination of 26 mycotoxins, including both well-known and emerging mycotoxins, in durum wheat was proposed by Juan et al. . AccuracFusarium mycotoxins in processed grains was described by Kai et al. [The simultaneous detection of five types of i et al. using anin house according to the criteria of National Public Health Laboratory\u2019s standard operating procedure No: A03-005 for Method Validation in Chemical Analysis and then applied to the analysis of 25 commercial vegetable oil samples from Malaysia. Another paper, published by Sharmili et al. , describA simple, rapid and accurate method based on QueEChERS followed by SPE C18 columns and detection with UPLC\u2013MS/MS was optimized and validated for the simultaneous determination of 25 mycotoxins in cereals . The optAnother method was developed for the simultaneous determination of 21 mycotoxins in white peony root, Radix Paeoniae Alba (RPA), by using a QuEChERS-based approach followed by d-SPE (C18 sorbent) and detection by UHPLC Q-LIT MS . In partAlternaria toxins in wine, vegetable juices and fruit juices and presented for the first time analytical data for altenuic acid, altenuisol and iso-altenuene. Furthermore, an improvement of the chromatographic performance was achieved for tenuazonic acid compared to previously reported multi-analyte methods for Alternaria targeted toxins in foodstuffs. Validation results were compliant with the requirements reported in the CEN technical report on the performance criteria for single laboratory validated methods of analysis for the determination of mycotoxins [Alternaria toxins.Zwickel et al. reportedcotoxins . The valin house in terms of sensitivity, linearity and accuracy, and successfully applied to the analysis of targeted mycotoxins in 64 infant cereals samples from the U.S. market. Al-Taher et al. developeIn another study, a fast, easy and cheap method was developed for the determination of eight mycotoxins in cereal-derived products based on the use of QuEChERS extraction and LC-MS/MS analysis . PerformA method based on LC-MS/MS was developed and validated for the analysis of 11 mycotoxins in edible oils . A simplAnother example of multiclass mycotoxin analysis was proposed also for cow milk samples by Flores-Flores and Gonzalez-Penas . SpecifiFusarium mycotoxins were the most frequently and co-occurring detected mycotoxins in the investigated samples.Kim et al. developeAspergillus, Fusarium, and Penicillium fungi and 12 ergot alkaloids produced by Claviceps purpurea in wheat and maize. The simple extraction procedure based on the \u201csalting-out\u201d LLE, minimizing the epimerization of ergot alkaloids, in combination with UPLC-MS/MS analysis, were the key factors for the rapid, robust and sensitive detection of these targeted compounds. The method was successfully applied to study the co-occurrence of targeted mycotoxins in 28 wheat and corn samples collected from six European countries.An innovative approach to multi-class mycotoxin control was proposed by Arroyo-Manzanares et al. for the Fusarium mycotoxins analysis, including their modified forms, in beer has been described [The feasibility of using an ultra-high-performance supercritical fluid chromatography (UHPSFC) and MS/MS as separation tool for 15 escribed . The samIn another paper, Du et al. describead hoc SPE clean-up was optimized by comparing four different approaches with Mycospin 400 column enabling acceptable recoveries for all mycotoxins including ochratoxin A and sterigmatocystin, which showed lower recoveries in the other tested protocols. The application of the developed method on 80 maize samples collected from Shandong Province in China showed that more than 70% of samples were contaminated with at least one of targeted mycotoxins. Li et al. describeAnother application of UPLC-MS/MS was described for determination of citrinin and ochratoxin A in a variety of food and feed matrices . A QuEChin house on corn, rice and feed according to the EC Regulation 401/2006 [A comparison of different approaches commonly used for the LC-MS/MS analysis of 12 mycotoxins in cereal foods was carried out by Solfrizzo et al. . In part401/2006 . PerformA modified QuEChERS method combined with nano flow LC-HRMS was proposed by Alcantara-Duran et al. for the Dong et al. describeFusarium and Aspergillus mycotoxins, including emerging and masked mycotoxins. Specifically, the dilute-and-shoot and SPE clean-up approaches were compared in combination with LC-MS/MS analysis. Results, in terms of precision, accuracy and reliability, indicated that both methods, thanks to the reduction of time and cost of the analysis, were promising for high throughput routine multimycotoxin analysis. It was the first time that Oasis\u00ae PRiME HLB columns were applied for the clean-up of the targeted mycotoxins considered in the study.Another comparative study was carried out by Scarpino et al. that devRecently, Woo et al. describeAn LC-MS/MS method for the simultaneous determination of seven major trichothecenes in wheat, wheat flour and wheat crackers was recently validated by a collaborative study involving 15 participant laboratories . This stThe necessity to monitor mycotoxins also in dried seafood products has recently led to the development of a sensitive, selective and accurate LC-MS/MS for the quantification of aflatoxin B1, T-2 toxin, ochratoxin A and deoxynivalenol in these specific matrices . After cGbashi et al. proposedVery recently a rapid and sensitive QuEChERS-UPLC-QTOF method, based on matrix-matched calibration, was developed and validated for the determination of 17 mycotoxins, including emerging ones, in malted barley and beer . ValidatA dispersive liquid\u2013liquid microextraction in combination with LC-MS/MS analysis was described for the simultaneous determination of 12 mycotoxins in rice bran . The metFinally, a recent study has proposed a multi-mycotoxins immunoaffinity column (multi-IAC) and LC-MS/MS method to evaluate 10 mycotoxins in traditional Chinese medicinal materials (TCMMs) . The metBoraginaceae, Asteraceae, and Legumionsae (Fabaceae) and about half of them affect wildlife, livestock and humans. PAs have also been evaluated as undesirable substances in food and feed by the EFSA [Brassicaceae, Solanaceae and Erythroxylaceae. The (-)-enantiomers hyoscyamine and scopolamine are the most studied TAs which, in contrast to the (+)-enantiomers, are naturally formed. The racemic mixture of (-)-hyoscyamine and (+)-hyoscyamine is called atropine [Datura stramonium, EFSA concluded that there is no information available on the carry-over of TAs from feed into animal products, such as milk or tissues from exposed animals, except for traces of alkaloids that have been found in eggs [Alkaloids are a group of amino-acid-derived and nitrogen-bearing molecules produced by several plant species that serve as a natural defense against aggression from other organisms such as insects or herbivores . Alkaloithe EFSA . The toxthe EFSA ,77,79. Bthe EFSA . In the atropine . Althoug in eggs conclude in eggs . Further in eggs . In the in eggs .n) or differential ion mobility mass spectrometry can be considered very helpful in discriminating isobaric compounds [Analytical methods for pyrrolizidine alkaloids (PAs) and N-oxide derivatives are typically based on GC-MS or LC-MS determinations. GC-MS methods commonly require the reduction of N-oxide derivatives and a subsequent derivatization step prior to chromatographic analysis. Other methods quantify total PAs by reducing PAs to necine pyrroles without any structural discrimination. On the other hand, the use of LC-MS analysis permits the quantification of both PAs and relative N-oxides without reduction steps ,86. Clasompounds . In Juneompounds .Datura species by LC-MS. The use of a simple sample preparation step , together with the use of a new generation of core-shell particle packed LC column, made the method simple and fast. The method was validated in house and applied to determine the targeted toxins in dry-plant materials of four different Datura species, with higher contamination levels in samples collected in autumn.Jakabova et al. describeA method allowing the detection of 11 pyrrolizidine alkaloids in honey by LC-ITMS was described by Griffin et al. . An SPE Mudge et al. proposedPrzewalskia tangutica Maxim. fruit extracts, a medicinal plant found in the Tibetan Plateau of China [An innovative, fast and specific method was described for the determination of anisodine, scopolamine, anisodamine and atropine by LC-MS/MS in of China . The autAnother application to honey was reported by Lorena et al. for the Valese et al. developeThe determination of atropine and scopolamine in buckwheat and derived products, soy, wheat, millet and chia seeds by UHPLC-ESI-HRMS has been described . The cleIn the same year, Martinello et al. describe3), and MRM with differential ion mobility spectrometry (DMS) were evaluated and compared in terms of selectivity and applicability. The DMS+MRM mode provided better results. The method using isotopically labeled PAs was validated according to the EURACHEM Guide [In another study, a UPLC-MS/MS analytical method was proposed for the determination of 15 PAs and 13 respective N-oxides in different groups of food . The ultEM Guide and showCirlini et al. describeThe development and validation of an LC-MS method for the determination of 10 PAs in honey was described by Kowalczyk and Kwiatek . After cRecently, an LC-HRMS method, based on SPE purification (with SCX sorbent), was described by Hungerford et al. for screG. procumbens and 7 commercial finished products. Results indicated for the first time that the occurrence of the targeted toxins in this plant material at variable levels might be depending on the geographical origin of the herb. The use of LC-HRMS was also described by Ji et al. for the Wang et al. developeIn another study, Zheng et al. describeBasle et al. describeA novel LC-MS/MS method was developed for the simultaneous determination of 21 TAs and 33 PAs together with their N-oxides in plant-based food matrices, such as sorghum, oregano, and mixed herbal tea using d-SPE clean-up . All thein house in black tea, peppermint and fennel and then two proficiency tests (PTs), addressing the determination of targeted TAs, were performed. This was the first time in which a subset of PTs participants strictly followed the specified analytical protocol provided by the coordinator and these data were additionally exploited to derive interlaboratory performance characteristics indicating that the method was a good candidate for standardization.Recently, the European Commission\u2019s Joint Research Centre proposed an LC-MS/MS analytical method for the determination of atropine and scopolamine in cereal-based foods for infants and young children, tea and herbal infusions . The metKaczynski and Lozowicka describeA simple and sensitive method using HILIC-MS/MS was developed for the determination of atropine and scopolamine in honey . A samplTussilago farfara and Lithospermi erythrorhzion [Recently, an LC\u2013MS/MS method coupled with 50% methanol extraction and MCX-SPE purification was developed for the determination of 28 PAs in two herbal medicines, rorhzion . The anarorhzion . The devAlexandrium, Gymnodinium, Dinophysis, and Pseudo-nitzschia. Marine toxins can accumulate in the tissue of filter feeding bivalves, including mussels, clams, scallops and oysters. On the basis of their poisoning symptoms, these toxins are classified as paralytic shellfish poisoning, amnesic shellfish poisoning, diarrhetic shellfish poisoning, neurotoxic shellfish poisoning, and ciguatera fish poisoning. Furthermore, marine biotoxins have different solubility and can be hydrophilic, including domoic acid, or lipophilic toxins. Major lipophilic toxins include okadaic acid, dinophysis-toxins, azaspiracids, pectenotoxins, yessotoxins and cyclic imines, which include spirolides, pinnatoxins, pteriatoxins and gymnodimines. Harmful algal bloom (HAB) is a natural phenomenon caused by the overgrowth of marine phytoplankton under certain environmental conditions. Specifically, this phenomenon has been increasing due to rising ocean temperature and growing coastal eutrophication . About 3For many years, mouse bioassays (MBA) have been used as reference methods to test for the presence of marine biotoxins in the food items, mainly for lipophilic and paralytic shellfish poisoning toxins. However, MBA tests show lack of specificity, poor sensitivity, false positives and are time-consuming, in addition to ethical concerns. Therefore, MBA and other biological methods have been used only for the detection of new or unknown toxins and not for routine analysis . SeveralThree papers focused on the use of LC-MS for the analysis of domoic acid in shellfish. In the first paper, an automated method based on the use of LC\u2013MS/MS for the determination of domoic acid in shellfish was described by Regueiro et al. . For thein house also using CRMs. Optionally, the use of an information-dependent acquisition approach in combination with the developed method provided an extra level of confirmation by library searching of product ion spectra also including unknown and unexpected isomeric analogues. The proposed method was applied to toxic mussel samples collected from western Canada in 2011, following an emergency, and highlighted the presence of high levels of dinophysistonin-1.Blay et al. describeA single-laboratory validation of a HILIC-MS/MS method was described by Turner et al. for the in house and applied to the analysis of 100 marine organisms including puffer fish, shrimps, crabs, clam and horseshoe crab collected from Chinese local markets with about 10% of samples contaminated with tetrodotoxin.Zhang et al. describeThe development of a HILIC-MS/MS method for the determination of 14 paralytic shellfish toxins in oysters, greenshell mussels and dinoflagellate cultures was described by Thomas et al. . The metAnother study aimed to develop and validate a HILIC-MS/MS method for the detection of eight paralytic shellfish poisoning toxins in shellfish was reported by Yang et al. . For the\u00ae and C18 cartridges. Then, three different chromatographic conditions were compared for both LC-MS/MS and LC-HRMS analysis in order to investigate the selectivity of the targeted compounds. Although LC-MS/MS permitted an optimal sensitivity in agreement with EFSA and US guidelines, LC-HRMS allowed the identity confirmation of Pacific-ciguatoxins analogues.Sibat et al. describeRecently, a screening method for detection of anatoxin-a, cylindrospermopsin, saxitoxin, nodularin and microcystins in fish tissue was described by Haddad et al. . AnatoxiRecently, an ultra-performance HILIC\u2013MS/MS was used for the screening of a total of 17 toxins, including paralytic shellfish toxins and tetrodotoxins, in bivalve molluscan species collected in Sweden . SimilarSolanaceae family, which contribute to plant resistance against pests and pathogens. Within Solanacee plants, the highest levels of GAs are found in potatoes, eggplants and tomatoes, even though the glycoalkaloids of most relevance to food safety, specifically \u03b1-solanine and \u03b1-chaconine, are those occurring in the potato . Several factors including mechanical damage to the plant, adverse storage and processing conditions may produce a significant increase of GAs content in potatoes [Glycoalkaloids (GAs) are natural toxic secondary metabolites commonly found in the plants of the potatoes . In humapotatoes . Howeverpotatoes ,137. Thepotatoes . To datepotatoes .The determination of GAs can be carried out by using different confirmatory methods based on GC-MS, HPLC coupled with UV-vis detector or with MS or MS/MS. The use of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) has been also reported. Enzyme-linked immunosorbent assay (ELISA) methods have been also described for GAs screening in food commodities . Nowadays, HPLC methods have replaced GC methods; in particular, LC-MS based methods are increasing their popularity in the analysis of GAs due to their higher selectivity and sensitivity compared to traditional detectors. Methods based on LC-MS/MS are commonly used for targeted analysis of GAs, while methods using LC-HRMS, mainly with Orbitrap technology, are more frequently used for metabolomic studies and to detect unknown compounds. To reduce interferences of complex matrices efficient sample pretreatment methods including ultrasonic extraction, pressurized liquid extraction and SPE have been used ,138. in house validated method was applied to screen 10 Irish potatoe samples, also evaluating if the cooking procedure affected the GAs content. Levels of GAs were higher in the skin than in the flesh, with concentrations lower in fried potatoes than those in boiled ones.In the first paper, Caprioli et al. describeThree papers describing the use of LC-MS based methods for the determination of \u03b1-solanine and \u03b1-chaconine in potato and derived products are described in the present review. Firstly, Liu et al. developeLelario et al. describeSolanum scabrum and S. nigrum berries from Kenya and US using UHPLC-MS/MS. Specifically, for the first time, the use of in-source fragmentation prior to the MS/MS analysis was applied as pseudo-MS to transform the GAs glycosides into the relevant aglycones. This approach permitted overcoming analytical issues due to the complexity and diversity of glycosides, as well as the limited availability of reference standards. Another method was proposed by Lyu et al. for the in house validated and applied to seven different protein isolates of relevance to the food industry to investigate the role of washing with water on GAs removal, and the effect of storage time on GAs level in protein from potato tubers following harvest. The results showed that total GAs increased during the storage of the potatoes.Finally, a study reported the development of an LC-ESI-MS method for the quantification of \u03b1-solanine, its aglycon form solanidine, and the \u03b1-chaconine in potato protein isolates . After aRutaceae , Leguminosae and Umbelliferae families. Furocoumarins have phototoxic properties and are produced in response to stress as defense against viruses, bacteria, fungi, insects and animals and are classified as natural pesticides [Furocoumarins are natural toxins found mainly in edible plants belonging to the sticides . On plansticides ,148. Dessticides . UHPLC hA rapid UHPLC-MS method was described for the determination and quantitation of 21 furocoumarins and 6 coumarins in citrus peel after extraction with a methanol/water mixture . The optRadix Angelicae Pubescentis (RAP) and its related preparations was described for the first time by Li et al. [In another paper, the use of an LC-MS/MS method for simultaneous determination of nine furocoumarin and two coumarins of i et al. . The optin house validated method were suitable for the extraction of furocoumarins from the investigated samples. Specifically, all toxins were detected in 25 foods with fresh parsley containing the highest content of bergamottin, followed by bergapten and 6\u20327\u2032-dihydroxybergamottin. The described study enabled a more accurate estimation of dietary furocoumarin exposure and was useful for epidemiological works investigating the relationships between furocoumarin intake and health outcomes. Furthermore, the authors compiled a database for furocoumarins representing the most comprehensive information on these toxins in popular foods and beverages available. Two papers described the development of UPLC-MS/MS methods for the determination of bergaptol, psoralen, 8-methoxypsoralen, bergapten, 6\u20327\u2032-dihydroxybergamottin, epoxybergamottin and bergamottin in American food by UPLC-MS/MS and using QuEChERS method for furocoumarins extraction. Firstly, Lee et al. applied Notopterygii Rhizoma et Radix (NRR), an important constituent of traditional Chinese medicine [The use of UHPLC-MS/MS was applied to the analysis, for the first time, of 15 furocoumarins, 8 coumarins and 7 phenolic acid esters in medicine . For optFinally, Arig\u00f2 et al. reportedFabaceae, Rosaceae, Leguminosae, Linaceae and Compositae, while linamarin, linustatin, neolinustatin, lotaustralin, taxiphyllin, amygdalin, dhurrin and prunasin are the main CNGs that have received considerable attention from EFSA, due to their frequent occurrence in agri-food products [Cyanogenic glycosides (CNGs) are secondary plant metabolites present in more than 2600 species and are produced as a chemical defense response to herbivores and pathogens after tissue damage. Indeed, CNGs are hydrolyzed to cyanohydrins and hydrocyanic acid (HCN) upon contact with plant endogenous \u03b2-glycosidase following maceration or wounding of the plant, or by gut microflora, following ingestion of the plant material ,156. Theproducts . CNGs arproducts . Other fproducts . These fproducts . Acute aproducts . Moreoveproducts ,161,162.products a maximuproducts . Furtherproducts .Quantification of CNGs in plant-based food can be made directly or indirectly. Specifically, direct methods are based on determining the intact glycoside in the sample and the few available methods are mainly based on HPLC analysis combined with UV-DAD or MS detection, with the latter ones improving sensitivity and selectivity. Less frequently, GC-MS methods, producing acceptable results in terms of high resolving power and automation, and ELISA assays, have been applied to quantify CNGs . The indThree papers described the use of LC-MS-based methods for the determination of amygdalin in almonds. In general, amygdalin is used as an indicator of the bitterness of almonds. Bitter almonds contain high levels of amygdalin which provides a characteristic cyanide aroma with moisture; non-bitter varieties contain trace levels of amygdalin and present nutty flavors. Finally, semi-bitter almonds have a \u201cmarzipan-like\u201d taste. Specifically, Toomey et al. describeThe other two papers described the determination of linustatin and neolinustatin in flaxseeds. The first application was reported by Wang et al. , which uGunasekera et al. reportedRecently, Tanaka et al. describeIn another recent paper by Zhong et al. , the UHPFinally, an UHPLC-MS/MS method was developed and validated for the presence of amygdalin, dhurrin, prunasin and linamarin in American elderberry fruit . The optThe LC-MS technology, either using triple quadrupole or high-resolution approaches, is the best and suitable approach in terms of selectivity and sensitivity for the simultaneous determination of multiclass analytes in such complex food matrices. Most of these methods use generic and rapid extraction protocols such as dilute and shoot and QuEChERS to cover as much as possible a wide range of analytes. Wang et al. proposedIn another paper, Lee et al. describeA multi-class UHPLC-MS/MS method for the determination of beauvericin, enniatins A and B, enniatins A1 and B1, and cereulide in corn, wheat, pasta and rice was reported by Decleer et al. . A fast Three papers were focused on the development and validation of a multiclass method for the determination of mycotoxins and TAs in cereals and derived products. In the first paper, an LC-MS/MS method was developed and validated for the simultaneous determination of 20 ergot alkaloids (mycotoxins) and six TAs in 113 grain-based foods for infants and young children . The proAn LC-MS/MS-based multi-toxin method was applied to the quantitative determination of mycotoxins and CNGs in cassava samples . The metDanezis et al. reportedIn a more recent study, the use of two-dimensional (2D) LC-MS/MS was reported for the first time for the determination of three classes of natural toxins, mycotoxins, tropane alkaloids and pyrrolizidine alkaloids, together with pesticides and growth regulators in oats and whole grains . A simplIn the same year, Zhao et al. proposedFinally, another 2D-LC-MS/MS multiclass method was developed and validated for the determination of mycotoxins and TAs together with plant growth regulators and pesticides in cereals . A rapidMass spectrometry-based methods (LC-MS) have become an essential tool for the monitoring of natural toxins as well as other chemical contaminants in food in order to ensure the safety of products, preserving consumer\u2019s health. In this paper, we have reviewed the literature from 2011 to 2021 on the use of LC-MS for the detection of natural toxins in food. Specifically, the review focuses on the following six classes of natural toxins: mycotoxins, alkaloids, marine toxins, glycoalkaloids, cyanogenic glycosides and furocoumarins. The highest number of papers was collected for mycotoxins, therefore for this class the more restricted period from 2016 to 2021 was considered. Generally speaking, the most used sample preparation approaches included solid liquid extraction, followed by solid phase extraction column clean-up, or alternatively QuEChERS approaches. For the separation and detection mode, UHPLC is mainly used as an improved approach of chromatographic separation while MS/MS methodology is the current leading approach for detection, mainly for the analysis of known compounds. Specifically, the use of matrix-matched calibration curves and isotopically labelled standards allow compensating matrix effects, improving the accuracy of the methods. However, the use of LC-HRMS has become increasingly common for routine analysis. Indeed, thanks to the untargeted data acquisition, retrospective analysis can be carried out, allowing to screen and quantify also parent and unknown compounds as in the case of unexpected compounds. The LC-HRMS advance is mainly due to the capability of this technique to separate with high accuracy and sensitivity also closely related compounds, such as isomers. Hybrid instruments combining two different types of analyzers, mainly Q-TOF and Q-Orbitrap, are commonly applied to improve the selectivity and sensitivity of these analytical methods. Although the majority of available methods for the detection of natural toxins in food are in-house validated according to national or international protocols, much effort is still necessary to assess the suitability of published LC-MS methods for their standardization through interlaboratory validations according to international guidelines . Recently, a clear trend in this sector is towards the development of new LC-MS methods capable of monitoring molecules showing different physicochemical properties, as in the case of natural toxins belonging to different classes, multicontaminants or multianalytes . The 2D-LC-MS/MS represents an alternative approach that is being successfully applied for the separation of multiclass analytes occurring in food complex matrices. Another emerging and interesting trend is represented by the application of HRMS using innovative untargeted metabolomics approaches to the screening of food samples without any prior information on the investigated analytes. As this approach is still challenging, efforts towards developing computational tools are necessary together with an appropriate implementation, validation and dissemination of databases for a wide range of analytes."} +{"text": "N/WC-Co can be improved when the coating and coating/matrix are doped separately with the main surface of intrinsic graphene or single vacancy defective graphene. Furthermore, the model electronic structures are analyzed. The results show that there exist strong Si/Co and N/Co covalent bonds in the interfaces when the matrix is doped with the main surface of intrinsic graphene, which causes the adhesion work of TiAlSiN/WC/msGR/Co to be greater than that of TiAlSiN/WC-Co. Additionally, when the graphene is doped into the coating, in the interface of TiAlSiN/msGR/TiAlSiNN/WC-Co, there exist strong N/Co covalent bonds that increase the interface adhesion work. Additionally, more charge transfer and orbital hybridization exist in the coating/matrix interface doped with the main surface of intrinsic graphene or single vacancy defective graphene, which explains the essential mechanism that the adhesion work of TiAlSiNN/msGR/WC-Co is greater than that of TiAlSiNN/WC-Co, and the adhesion work of TiAlSiNN/svGR/WC-Co is greater than that of TiAlSiNN/WC-Co.Based on the first-principles method, TiAlSiN/WC-Co interface models with graphene doped into the matrix, coating, and the coating/matrix are constructed. The interface adhesion work is calculated and modeled to study the interface bonding properties from the atomic microscopic point of view. The results show that the interface bonding properties of TiAlSiN/WC-Co can be improved when the matrix is doped with the main surface of intrinsic graphene, and the interface bonding property of TiAlSiN Modern machining technologies put forward higher requirements for cutting tools . TraditiBy using the PVD method, the TiAlSiN-coated cemented carbide tool is fabricated by depositing TiAlSiN coating on the cemented carbide WC-Co matrix, which has high hardness, wear resistance, and oxidation resistance. Qiang Chen et al. , Yan Wu Graphene (GR), as a nanomaterial, has been applied in cutting-tool materials because of its special structure and properties ,10. XiaoCurrently, research on TiAlSiN-coated carbide tools mainly focuses on experimental aspects. There is less research on TiAlSiN-coated carbide tools that concerns the effect of graphene doping, from an atomic microscopic point of view, on the interface bonding property of this kind of tool coating. Therefore, using the first-principles method, we report the interface bonding properties revealed by graphene-doped TiAlSiN/WC-Co interface models in which graphene is doped into cemented carbide WC-Co matrix, TiAlSiN coating, and the coating/matrix interface. The research results are of great significance for the interface optimization design of TiAlSiN-coated carbide tools.The geometry optimization of the WC, Co, GR, and TiAlSiN crystal models is performed to obtain the lowest energy and most stable lattice system.\u22125 eV/atom [All calculations are based on spin-polarized density functional theory (DFT) using the CASTEP module ,26,27. T eV/atom .After setting the above parameters, the geometry optimization of WC, Co, GR and TiAlSiN crystal models is carried out. The lattice cell parameters of WC, Co, GR and TiAlSiN after geometry optimization are shown in The interface models are constructed by the following steps:(1) The WC(0001) crystal face with the W atom as the terminal is the most stable , and the(2) Si atoms in TiAlSiN existed in the form of replacing Ti atoms at the interface , and TiAAl/WC-Co, TiAlSiNSi/WC-Co, and TiAlSiNN/WC-Co without graphene doping were built, as shown in The interface bonding strength between the TiAlSiN coating and WC-Co matrix directly affects the service life of coated carbide tools. There are three kinds of terminal atoms on the surface of the TiAlSiN crystal. Therefore, the interface models of TiAlSiNSi/WC-Co has the strongest interface bonding property, and TiAlSiNN/WC-Co has the weakest interface bonding property. Therefore, based on the interface models of TiAlSiNSi/WC-Co and TiAlSiNN/WC-Co, the effect of graphene doping on the interface bonding property of TiAlSiN/WC-Co was studied.Shulei Li et al. found thSi/WC-Co and TiAlSiNN/WC-Co interface models, the models of matrix doped with intrinsic and defective graphene, respectively, of TiAlSiN/WC-Co are constructed. The preferred orientation of graphene is (001). The most representative single-layer graphene is selected to construct a single-layer GR(001) surface, and a 10 \u00c5 vacuum layer is added to form the GR(001) interface model, which is placed between WC and Co in the WC(0001)-Co interface model. The orientation relationship is TiAlSiN(111)/WC(0001)/GR(001)/Co.(3) Graphene mainly includes intrinsic graphene and defective graphene . Intrins(4) Similarly, in order to study the effect of doping graphene into the coating and coating/matrix interface on the interface bonding property of TiAlSiN/WC-Co, the GR(001) interface model is placed in the TiAlSiN interface model to construct models of the coating doped with intrinsic and defective graphene. The orientation relationship is TiAlSiN(111)/GR(001)/TiAlSiN(111)/WC(0001)-Co. The GR(001) interface model is placed between the coating TiAlSiN and matrix WC-Co, and the models of coating/matrix interface doped with intrinsic and defective graphene of TiAlSiN/WC-Co are constructed. The orientation relationship is TiAlSiN(111)/GR(001)/WC(0001)-Co.The construction process of graphene-doped TiAlSiN/WC-Co interface model is shown in Si/WC-Co and TiAlSiNN/WC-Co models, interface models of matrix doped with intrinsic graphene are constructed, as shown in Si/WC/msGR/Co, TiAlSiNSi/WC/acGR/Co, and TiAlSiNSi/WC/zzGR/Co combining the matrix with the main surface of intrinsic graphene and the graphene boundary are constructed, as shown in N/WC/msGR/Co, TiAlSiNN/WC/acGR/Co, and TiAlSiNN/WC/zzGR/Co are constructed, as shown in Specifically, based on the TiAlSiNSi/WC/svGR/Co, TiAlSiNSi/WC/tdGR/Co, TiAlSiNN/WC/svGR/Co, and TiAlSiNN/WC/tdGR/Co, with defective graphene doped matrix are constructed, as shown in Four interface models, TiAlSiNWith the same method, the interface models of coating and coating/matrix interface doped with graphene are constructed, as shown in The adhesion work can characterize the bonding strength of the interface structure. The greater the adhesion work, the stronger the interface bonding property and the more stable the interface structure. Furthermore, the charge-density difference and the density of states are analyzed by calculating the electronic structures of the model interface atoms, revealing the essence of interface bonding from the perspective of charge transfer and bonding mode.\u03b1/\u03b2 adhesion work (Wad) calculation formula is [adW is the adhesion work, J/m2; \u03b1E is the total energy of surface \u03b1, eV; \u03b2E is the total energy of surface \u03b2, eV; \u03b1/\u03b2E is the total energy of the interface system, eV; and the total interface area is given by A, \u00c52.The interface rmula is :(1)Wad=ESi/WC/msGR/Co is larger than that of TiAlSiNSi/WC-Co without graphene doping, and the interface adhesion work of TiAlSiNN/WC/msGR/Co is larger than that of TiAlSiNN/WC-Co, which indicates that the combination of the matrix and the main surface of intrinsic graphene improves the interface bonding property of TiAlSiN/WC-Co. When the matrix is doped with the graphene boundary, the adhesion works of TiAlSiNSi/Co interface and TiAlSiNN/Co interface are less than those without doping, which indicates that the combination of matrix and graphene boundary reduces the interface bonding property of TiAlSiN/WC-Co.The calculated interface adhesion works of matrix doped with intrinsic graphene are shown in Si/Co and TiAlSiNN/Co are less than those without doping, respectively, which indicates that the combination of matrix and defective graphene reduces the interface bonding property of TiAlSiN/WC-Co.The calculated interface adhesion works of matrix doped with defective graphene are shown in By comparing adhesion works of Si and N terminal models of matrix doped with graphene, given in Si/Co interface of TiAlSiN/msGR/TiAlSiNSi/WC-Co is less than that of the TiAlSiNSi/Co interface of TiAlSiNSi/WC-Co without graphene doping, and the interface bonding property of TiAlSiN/msGR/TiAlSiNSi/WC-Co decreases.The calculated interface adhesion works of coating doped with intrinsic graphene are shown in N/Co interface of TiAlSiN/msGR/TiAlSiNN/WC-Co are larger than that of the TiAlSiNN/Co interface of TiAlSiNN/WC-Co without graphene doping, and the interface bonding property of TiAlSiN/msGR/TiAlSiNN/WC-Co increases. The results indicate that the coating is doped with the main surface intrinsic graphene in the N terminal model, and the interface bonding property of TiAlSiN/msGR/TiAlSiNN/WC-Co increases. When the coating is doped with the graphene boundary, the adhesion works of TiAlSiNSi/Co and TiAlSiNN/Co are less than those without doping, which indicates that the combination of coating and graphene boundary reduces the interface bonding property of TiAlSiN/WC-Co.The adhesion works of the TiAlSiNN/WC-Co is doped with svGR, the adhesion work of each interface increases compared with that of TiAlSiNN/WC-Co without doping, and the interface bonding property increases. When the coating is doped with svGR, in the Si terminal model of TiAlSiNSi/WC-Co and different terminal models of doping tdGR, there are interfaces whose adhesion works decrease, and the interface bonding property decreases.The calculated interface adhesion works of coating doped with defective graphene are shown in By comparing adhesion works of Si and N terminal models doped with graphene in coating, given in Si/msGR/WC-Co of the Si terminal model is less than that of the TiAlSiNSi/WC-Co interface without doping graphene, and the interface bonding property of TiAlSiNSi/msGR/WC-Co decreases. The interface adhesion work of TiAlSiNN/msGR/WC-Co of the N terminal model is larger than that of the TiAlSiNN/WC-Co interface, and the interface bonding property of TiAlSiNN/msGR/WC-Co increases. When the coating/matrix interface is doped with the graphene boundary, in different terminal models, there are interfaces whose interface adhesion works are less than that of the TiAlSiN/WC-Co interface. It indicates that when the coating/matrix interface is doped with an intrinsic graphene boundary, the interface bonding property of TiAlSiN/WC-Co decreases.The calculated interface adhesion works of the coating/matrix interface doped with intrinsic graphene are shown in N/WC-Co is doped with svGR does the adhesion work of each interface increase compared with that of TiAlSiNN/WC-Co without doping, and the interface bonding property increases. In TiAlSiNSi/svGR/WC-Co, TiAlSiNSi/tdGR/WC-Co, and TiAlSiNSi/tdGR/WC-Co, there are interfaces whose interface adhesion works decrease, and the interface bonding property decreases.The calculated interface adhesion works of the coating/matrix interface doped with defective graphene are shown in By comparing adhesion works of Si and N terminal models doped with graphene in coating/matrix interface, given in After calculation, the adhesion works of the graphene-doped TiAlSiN/WC-Co interface model were summarized, as shown in N/WC-Co, while other forms of graphene doping cannot improve the interface bonding property of TiAlSiN/WC-Co. When the coating/matrix interface is doped with graphene, msGR and svGR improve the interface bonding properties of TiAlSiNN/WC-Co, while other forms of graphene doping cannot improve the interface bonding property of TiAlSiN/WC-Co.It can be seen from Si/WC-Co increases; when msGR is doped into the matrix, coating, and the coating/matrix interface of N terminal models, the interface bonding property of TiAlSiNN/WC-Co increases, but acGR, zzGR, and tdGR cannot improve the interface bonding property of TiAlSiN/WC-Co.From the perspective of different forms of graphene doping, it is found that when msGR is doped into the matrix of Si terminal models, the interface bonding property of TiAlSiN(1) Charge-density differenceSi/WC-Co and TiAlSiNN/WC-Co interface models is calculated, and the charge-density difference images are shown in The charge-density distribution in atoms can be analyzed by the charge-density difference image. The charge-density difference of TiAlSiNSi/WC-Co interface, there is charge accumulation between Si atoms and Co atoms, that is, covalent bonds exist; when the matrix is doped with msGR, this introduces msGR/Co and /WC/msGR into the interfaces. As shown in Si/WC/msGR/Co, and the electron cloud near the Co atom moves toward the Si atom, which enhances the attraction of Si to the Co atom and forms a strong Si/Co covalent bond. As shown in N/WC-Co interface indicates the existence of covalent bonds. As shown in N/WC/msGR/Co, N atoms are in the electron-enriched state, while Co atoms are in the electron-absent state, and there is attraction between N and Co atoms. When the matrix is combined with msGR, the newly introduced interface is stable and the original interface is well combined. In the four interface models, in which the matrix is doped with the graphene boundary, as shown in Si/WC/svGR/Co, TiAlSiNSi/WC/tdGR/Co, TiAlSiNN/WC/svGR/Co, and TiAlSiNN/WC/tdGR/Co.Si/WC-Co, shown in N/WC-Co, shown in Si/WC-Co, and the interatomic force is weak. As shown in Si/WC-Co. In N/WC-Co, and the interatomic force is strong. In N/WC-Co, and the interatomic force is strong. In other cases, the doping of defective graphene weakens the bonding force between atoms at the interface.Si/msGR/WC-Co interface, there are charges transferred between C atoms of msGR and Al atom, and there are shared charges between C and Si atoms. The charge density between Co and C atoms is thin, and the interatomic force is small. At the interface of TiAlSiNN/msGR/WC-Co, there are obvious shared charges between C atoms of msGR and N, Co atoms, and the interatomic force is strong. When the coating/matrix interface is doped with the graphene boundary, the combination of coating/matrix interface and graphene boundary makes the charge between atoms thin. The interatomic force decreases, and the interface bonding property decreases. Compared with the N terminal model, when the coating/matrix interface is doped with msGR and zzGR, there are more electronic blank areas at the interface of the Si terminal model, and the interatomic force is small, and the interface bonding property decreases. It can be seen that the interface bonding property decreases most obviously when the Si terminal model is doped with graphene, and the results are consistent with those given in Si/svGR/WC-Co, the charge density between Al, Si, and Co atoms is thin, there are charge blank areas between C and Co atoms, and the interatomic force is weak. In Si/tdGR/WC-Co, the charge density between Al, Si, and C atoms is thin. The charge density between Co and C atoms is thin and there are blank areas. In N/svGR/WC-Co, the electrons between C and N atoms and between C and Co atoms all overlap more, and the bonding force is strong. In N/tdGR/WC-Co, the charge density between C and Co atoms is thin, and the interatomic force is weak. Summarizing, only the atomic charges at the interface of TiAlSiNN/svGR/WC-Co overlap more, and the bonding force between atoms is strong. In other cases, the defective graphene doping weakens the bonding force between atoms at the interface, and the interface bonding property decreases.(2) Density of states (DOS)Analyzing the density of states can explore the bonding situation from the perspective of electron orbital hybridization.Si/WC-Co is that its peak is mainly contributed by p orbital electrons of Ti atoms and W atoms, and the interaction between Si and Co at the interface is mainly the hybridization of p electrons of Si and d electrons of Co, forming Si/Co covalent bonds. Si/WC/msGR/Co overlaps in the range \u22125~5 eV, and the orbital hybridization is generated by Al-s, Si-s, and Co-d, forming Al/Co bonds and Si/Co bonds. Compared with Si/WC/msGR/Co is higher than that of TiAlSiNSi/WC-Co.N/WC-Co is that its peak is mainly contributed by p orbital electrons of Ti and W atoms, and the interaction between N and Co at the interface is mainly the hybridization of p electrons of N and d electrons of Co, forming N/Co covalent bonds. In N/WC/msGR/Co interface, the density of states of N and Co atoms overlaps on the left side of Fermi level, and N-s and Co-d produce orbital hybridization, forming N/Co bonds. On the left side of Fermi level, the density of states of the C atom, Co atom, and W atom is orbitally hybridized by C-p with Co-d and W-d, forming C/Co and C/W bonds. Compared with N/WC/msGR/Co is greater than that of TiAlSiNN/WC-Co.A shown in Si/WC/tdGR/Co is low, below 0.2 eV, and the degree of orbital hybridization of Al-p, Si-p, and Co-d is weak. In N/WC/svGR/Co is narrow, resulting in low orbital hybridization of Co-d with C-p and N-p, and weak interatomic bonding force. In N/WC/tdGR/Co is low, and the interatomic bonding force is weak. This explains the essential mechanism that the interface adhesion work of TiAlSiN/WC-Co decreases when its matrix is doped with defective graphene.Si/WC-Co, the charges between Al, Si, and Co atoms are rearranged, and at Fermi level, Al, Si, and Co atoms at the interface produce orbital hybridization, forming covalent bonds. Compared with Si/WC-Co is higher than that of TiAlSiNSi/WC-Co. In N/WC-Co, N-s and C-s produce orbital hybridization in the range \u221222~15 eV, and N-p and C-p produce orbital hybridization in the range \u221210~3 eV, forming N/C bonds. Al-s, Al-p and C-s, and C-p produce orbital hybridization in the range \u221217~3 eV, forming Al/C bonds. N-p and Co-d produce orbital hybridization in the range \u22125~3 eV, forming N/Co bonds. The density of atomic states of forming chemical bonds and orbital hybridization degree is high, which explains the essential mechanism that the adhesion work of TiAlSiN/msGR/TiAlSiNN/WC-Co is greater than that of TiAlSiNN/WC-Co.Si/WC-Co overlaps in the range \u22126~3 eV, the N-p density of states is below 0.1 eV, and the energy is low, and the N/Co bond is weak. The density of states of Al and Si atoms is low, below 0.3 eV, and the degree of hybridization with Co-d orbitals is weak. In N/WC-Co interface, the charges between N and Co atoms are rearranged, and at the Fermi level, N and Co atoms at the interface produce orbital hybridization, and the degree of orbital hybridization is low, forming weak covalent bonds. In N/WC-Co, Co and N atoms produce orbital hybridization, the hybridization area is small, and the energy is low, and the Co/N bond is weak. This explains the essential mechanism for the decrease in adhesion work at the TiAlSiN/WC-Co interface when the coating is doped with an intrinsic graphene boundary.Si/WC-Co produce orbital hybridization in the range \u22124~3 eV, and the degree of hybridization is low, forming weak Co/Si bonds. In Si/WC-Co overlaps in the range \u22124~3 eV, where the density of states of Co is low, and the degree of orbital hybridization of Al-p, Si-p, and Co-d is weak. In N/WC-Co are wide, and the C/N, C/Al, and Co/N bonds are strong. In N/WC-Co is high, while the N-p density of states is low, and the degree of orbital hybridization of Co-d and N-p is weak, and bonding force is weak. Therefore, the interface adhesion work of TiAlSiN/svGR/TiAlSiNN/WC-Co is greater than that of TiAlSiNN/WC-Co without graphene doping; in other cases, doping defective graphene in the coating reduces the interface adhesion work of the model.Si/msGR/WC-Co, the density of states of Al and Si atoms overlaps with C atoms of msGR in the range \u221210~3 eV, and Al-s, Al-p, Si-p, and C-p produce orbital hybridization, forming Al/C and Si/C bonds. The density of states of Co and C atoms overlaps in the range \u22125~3 eV, and the density of states of C atoms in the overlapping region is very low, the region of density of states of Co is narrow, and the Co/C bond formed by the orbital hybridization of Co-d and C-p is weak, which explains the essential mechanism that the adhesion work of TiAlSiNSi/msGR/WC-Co is less than that of TiAlSiNSi/WC-Co. In N/msGR/WC-Co, the density of states of N and C atoms of msGR overlaps in the range \u221220~10 eV, and the N-s and C-s produce orbital hybridization. In the range \u221210~3 eV, N-p and C-p produce orbital hybridization, which forms strong N/C bonds. The density of states of Co and C atoms overlaps in the range \u22125~3 eV, and Co-d and C-p produce orbital hybridization, forming strong Co/C bonds, which explains the essential mechanism that the adhesion work of TiAlSiNN/msGR/WC-Co is greater than that of TiAlSiNN/WC-Co.Si/acGR/WC-Co, the Al-s, Al-p, Si-s, and C-p produce orbital hybridization in the range \u221210~3 eV, forming Al/C and Si/C bonds. The Co-d and C-p produce orbital hybridization, and the degree of hybridization is low, forming weak Co/C bonds. In Si/zzGR/WC-Co overlaps in the range \u22125~0 eV, the C-p density of states is low, and the C/Co bond is weak. In N/acGR/WC-Co, the density of states of C-p and N-p overlaps in the range \u22127~0 eV, the C-p density of states is low, and the N/C bond is weak. In Si/svGR/WC-Co, the density of Al, Si, and C atoms of svGR overlaps in the ranges \u221217~15 and \u221210~3 eV. Al-s, Al-p, Si-p, and C-p produce orbital hybridization, forming the Al/C and Si/C bonds. The density of states of Co and C atoms overlaps in the range \u22124~3 eV. Co-d and C-p produce orbital hybridization, and the degree of hybridization is low, and the Co/C bond is weak. In Si/tdGR/WC-Co, the density of Al, Si, and C atoms of svGR overlaps in the ranges \u221217~15 and \u221211~3 eV. Al-s, Al-p, Si-s, Si-p, and C-p produce orbital hybridization to form Al/C and Si/C bonds. The density of states of Co and C atoms overlaps in the range \u22125~4 eV, and Co-d and C-p produce orbital hybridization. The peak value of C-p in the overlapping region is low, and the Co/C bond formed is weak. In N/svGR/WC-Co, the density of states of N-s and C-s overlap in the range \u221215~8 eV. The density of states of N-p and C-p overlap in the range \u22127~3 eV, and the degree of electron orbital hybridization is high, and the N/C bond is strong. In N/svGR/WC-Co is greater than that of TiAlSiNN/WC-Co without graphene doping. In other cases, doping defective graphene in the coating/matrix interface reduces the interface adhesion work of the model. The analysis results of density of states are consistent with the analysis results of previous adhesion works, which reveals the interface bonding mechanism of graphene-doped TiAlSiN/WC-Co.The partial research results are consistent with the experimental results in references ,21. RefeSi/WC-Co and TiAlSiNN/WC-Co, the interface models of graphene-doped matrix, coating, and coating/matrix interface are established, the interface adhesion works are calculated with the first-principles method. The results show that the matrix doped only with msGR improves interface adhesion works of TiAlSiNSi/WC/msGR/Co and TiAlSiNN/WC/msGR/Co, and the interface bonding property of TiAlSiN/WC-Co improves. The coating doped with msGR and svGR improves adhesion works and interface bonding properties TiAlSiN/WC-Co. Additionally, the coating/matrix interface doped with msGR and svGR improves adhesion works and interface bonding properties of TiAlSiN/WC-Co.(1) Based on the most stable and unstable interface models of coated, cemented carbide tools of TiAlSiNSi/WC/msGR/Co to be greater than that of TiAlSiNSi/WC-Co, and the adhesion work of TiAlSiNN/WC/msGR/Co to be greater than that of TiAlSiNN/WC-Co. Additionally, there exists strong N/Co covalent bonds in the interfaces of TiAlSiN/msGR/TiAlSiNN/WC-Co, which lead to the adhesion work of TiAlSiN/msGR/TiAlSiNN/WC-Co to be greater than that of TiAlSiNN/WC-Co. Additionally, there exists more charge transfer and orbital hybridization in the coating/matrix interface doped with the main surface of intrinsic graphene and single vacancy defective graphene, which leads to the adhesion work of TiAlSiNN/msGR/WC-Co to be greater than that of TiAlSiNN/WC-Co, and the adhesion work of TiAlSiNN/svGR/WC-Co to be greater than that of TiAlSiNN/WC-Co.Based on these research results, the electronic structures of the models are further analyzed. The results show that there exists strong Si/Co and N/Co covalent bonds in the interfaces when the matrix is doped with the main surface of intrinsic graphene, which causes the adhesion work of TiAlSiNSi/WC/msGR/Co, there exists orbital hybridization between the Si p-electron and the Co d-electron, forming strong Si/Co covalent bonds. At the interface of TiAlSiNN/WC/msGR/Co, there exists the hybridization of the N p-electron and the Co d-electron, forming stable N/Co covalent bonds. When the matrix is doped with the graphene boundary, C atoms and Co and W atoms produce orbital hybridization, and the hybridization degree and the bonding between Al, Si, N atoms and Co atoms at the interface of TiAlSiN are weak. When the matrix is doped with the defective graphene, the combination of matrix and defective graphene changes the charge distribution at the interface, and the bonding between atoms at the interface of the model is weak.(2) Electronic structure analysis shows that when the matrix is doped with the main surface of graphene, msGR doping introduces msGR/Co and WC/msGR interfaces. At the interface of TiAlSiNSi/WC-Co, there are electronic blank areas between Al, Si, and Co atoms. At the Fermi level, there exists the orbital hybridization between Al, Si atoms, and Co atoms to form weak covalent bonds. At the interface of TiAlSiN/msGR/TiAlSiNN/WC-Co, N-p and Co-d produce orbital hybridization to form N/Co covalent bonds. When the coating is doped with the graphene boundary, the charge between atoms at the interface of the model is low, the hybridization degree between atoms at the interface is low, and the bonding is weak. When the coating is doped with the defective graphene, only the atomic charges at the interface of TiAlSiN/svGR/TiAlSiNN/WC-Co overlap more, and the degree of hybridization between atoms is high, and the interatomic force is strong. In other cases, the doping defective graphene weakens the bonding force between atoms at the interface.(3) When the coating is doped with msGR, in the model of TiAlSiN/msGR/TiAlSiNSi/msGR/WC-Co, and shared charges between C and Si atoms. The atomic charge density between Co and C atoms is thin, and the Co/C bond is weak, and the interatomic force is weak. At the interface of TiAlSiNN/msGR/WC-Co, there are obvious shared charges between C atoms of msGR and N and Co atoms. Co-d and C-p produce orbital hybridization to form strong Co/C bonds, and the interatomic force is strong. When the coating/matrix interface is doped with an intrinsic graphene boundary, the combination of coating/matrix interface and graphene boundary makes the charge between atoms thin, and the orbital hybridization degree between atoms decreases, and the interatomic force is weak. When the coating/matrix interface is doped with the defective graphene, only the atomic charges at the TiAlSiNN/svGR/WC-Co interface overlap more, and the orbital hybridization degree between atoms is high and the bonding force is strong. In other interface models, there exists weak electronic orbital hybridization, and the bonding force between atoms is weak. In other cases, doping defective graphene in the coating/matrix interface reduces the interface adhesion work of the models.(4) When the coating/matrix interface is doped with msGR, there exists charge transfer at the interface of TiAlSiN"} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "The internet of things (IoT) is an emerging paradigm of educational applications and innovative technology in the current era. While capabilities are increasing day by day, there are still many limitations and challenges to utilizing these technologies within E-Learning in higher educational institutes (HEIs). The IoT is well-implemented in the United States of America (USA), United Kingdom (UK), Japan, and China but not in developing countries, including Saudi Arabia, Malaysia, Pakistan, Bangladesh, etc. Few studies have investigated the adoption of IoT in E-Learning within developing countries. Therefore, this research aims to examine the factors influencing IoT adoption for E-Learning to be utilized in HEIs. Further, an adoption model is proposed for IoT-based E-Learning in the contexts of developing countries and provides recommendations for enhancing the IoT adoption for E-Learning in HEIs. The IoT-based E-Learning model categorizes these influencing factors into four groups: individual, organizational, environmental, and technological. Influencing factors are compared along with a detailed description in order to determine which factors should be prioritized for efficient IoT-based E-Learning in HEIs. We identify the privacy (27%), infrastructure readiness (24%), financial constraints (24%), ease of use (20%), support of faculty (18%), interaction (15%), attitude (14%), and network and data security (14%), as the significant E-Learning influencing factors on IoT adoption in HEIs. These findings from the researcher's perspective will show that the national culture has a significant role in the individual, organizational, technological, and environmental behavior toward using new technology in developing countries. Higher educational institutes (HEls) are vast intelligent systems. This is due to the fact that the main components of current HEIs, educators, and educated learners are all information-complex holographic humans provides massive opportunities for HEIs that bring together independent control and provision of better infrastructure robustness, scalability, and agility. The IoT allows humans and things to access from anywhere, anytime, and any place the link with anything and to any person without a specific path and service , and smart learning. Till now, the revolution of learning is divided into four groups including traditional, digital, E-Learning, and smart learning, as shown in Currently, we are living in the era of Smart learning. In simple words, Smart learning is the combination of E-Learning and IoT, or it is also known as IoT based E-Learning, as shown in Equation (1). In IoT based E-Learning, students learn from their educational institution or home with various IoT sources online. Similarly, instructors give the lectures in the form of different videos, meeting websites, and apps, while arranging the test also online by using the IoT or smart devices. Students feel the essence of physical classes in these virtual meetings or online courses Adoption in Higher Educational Institutions (HEIs) for E-Learning?\u201d\u201c* \u201cInternet of Things\u201d + \u201cIoT\u201d + \u201cHigher Educational Institutes\u201d + \u201cHEIs\u201d) and .This section presents the research steps followed to perform this study. According to Moher et al. , a comprIn the screening process, duplicate studies are removed at the first stage. After that, exclusion criteria have been applied based on the title, abstract, and body of the research article. The inclusion and exclusion criteria have also been applied on the basis of the study focusing on adopting new technology for E-Learning, written in English, and published in well-recognized journals and conferences. Moreover, content analysis has been performed to categorize the influencing factors, and a comprehensive list of E-Learning factors for IoT adoption in HEIs is compiled.This section explains the entire used system for IoT-based E-Learning and how it works and is addressed by existing researchers.In two universities in Saudi Arabia, a quantitative analysis is conducted with the help of 527 students. This analysis is shown that 96.7% of students are keen on adopting IoT technologies in the higher education system. In this analysis, two main factors, perceived usefulness and ease of use, are examined by using Technology Acceptance Model 3 (TAM3). Apart from other factors, the trust factor is also included in considering the importance of adopting IoT technologies in E-Learning based on systems of higher education systems is financially ready to pursue its objective. For example, IoT-based E-Learning necessitates a large number of tools and software and specialists to handle, all of which demand a large sum of money. The organization must consider these factors before deciding whether to implement an IoT-based E-Learning system. They must figure out how much it will cost in a month or a year. The organization must plan appropriately to make the most effective use of its funds. They must preserve all records, save money for future requirements, and be alerted to avoid needless spending while they spend encompasses hardware and software procedures, regulations, settings about network use, accessibility, and overall threat prevention. Access control, application security, firewalls, network analytics, several forms of network-related security , virus and antivirus software, VPN encryption, and more are all part of the network security. Defending client data and information, keeping shared data safe, maintaining dependable access and network performance, and protecting against cyber-attacks require network and data security. So, network and data security play an essential role in the IoT-based E-Learning in HEIs as the whole system needs to be protected from threats and risks and Universiti Teknologi Malaysia (UTM) for supporting this research through RUG grant No. IFRS 7873.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "ICD-11 was released by WHO in 2018 and approved by the World Health Assembly (WHA) in 2019. The revision for all chapters was guided by the principles of global applicability, scientific validity and clinical utility. The new chapter for mental health is termed 06 Mental, Behavioural or Neurodevelopmental Disorders (MBND). The ICD-11 with its chapter on Mental, Behavioural or Neurodevelopmental Disorders, its Mortality and Morbidity Statistics (MMS), Coding Tool and Reference Guide, Clinical Descriptions and Diagnostic Guidelines (CDDG), and other tools for translation and implementation offers an innovative approach for individualised diagnosis, treatment and care of people with mental disorders. For supporting the international process of implementation, WHO has installed an International Advisory Group for Training and Implementation of ICD-11 MBND. Development, Concept and Structure of ICD-11 will be presented. Selected changes from ICD-10 to ICD-11 like new diagnostic categories, revision of diagnostic criteria, introduction of dimensional symptom qualifiers or course descriptors, and options for complex coding with regard to their innovative strength, controversial potential and impact on diagnostics, treatment and care will be briefly discussed. National challenges for implementation - partly informed by international field trials, administrative, organisational, educational and training requirements - will be outlined. The new ICD-11 chapter on Schizophrenia or other primary psychotic disorders will serve as an example to discuss potential impact on treatment selection and outcome.No significant relationships."} +{"text": "Up-to-date, accurate information on the disease burden of motor neuron disease (MND) is the cornerstone for evidence-based resource allocation and healthcare planning. We aimed to estimate the burden of MND globally from 1990 to 2019, as part of the Global Burden of Disease, Injuries and Risk Factor (GBD) study. Amyotrophic lateral sclerosis, progressive muscular atrophy, primary lateral sclerosis, pseudobulbar palsy, spinal muscular atrophy and hereditary spastic paraplegia- were included for analysis as MNDs. We measured age-standardized incidence, prevalence, death, and disability-adjusted life-years (DALYs) in 204 countries and territories worldwide from 1990 to 2019 using spatial Bayesian analyses. The effects of age, sex, and the sociodemographic index on incidence, prevalence, death, and disability-adjusted life-years due to MNDs were explored. According to 2019 GBD estimates, there were ~268,673 prevalent cases and 63,700 incident cases of MND worldwide. In 2019, MND caused 1,034,606 DALYs and 39,081 deaths worldwide. The age-standardized rates of prevalence, incidence, death, and DALYs for MNDs in 2019 were 3.37 per 100,000 people, 0.79 per 100,000 people, 0.48 per 100,000 people, and 12.66 per 100,000 people, respectively. The global prevalence and deaths due to MND in 2019 were increased compared to 1990, without significant change in incidence. More than half of the prevalence and deaths due to MND occurred in three high-income regions . In most cases, the prevalence, incidence, and DALYs of MNDs were high in regions with high sociodemographic index; however, in high-income East Asia, these were relatively low compared to similar sociodemographic index groups elsewhere. The burden of MND increased between 1990 and 2019. Its expected increase in the future highlights the importance of global and national healthcare planning using more objective evidence. Geographical heterogeneity in the MND burden might suggest the influences of sociodemographic status and genetic background in various regions. Motor neuron diseases (MNDs) are rare neurological disease groups of neurodegenerative disorders associated with the degeneration of motor neurons in the upper and lower extremities . They inAlthough epidemiologic studies on MNDs have been published in the United States and Europe, their incidence, prevalence, and burden are not well known because the diseases are rare \u20136. AlthoThe purpose of this study was to investigate the global burden of MNDs, including the incidence, prevalence, death, disability-adjusted life-years (DALYs), years lived with disability (YLDs), and years of life lost (YLLs) between 1990 and 2019 from GBD information according to age, sex, regions, and the estimates from individual countries. Furthermore, we investigated the GBD of MNDs based on the sociodemographic index (SDI) reflecting the development of each country.The GBD Study is a systematic and comprehensive study of diseases worldwide. Based on the estimates of this study, it is possible to compare and analyze the current status of the global, regional, and national burden of diseases . The GBDhttps://vizhub.healthdata.org/gbd-compare/; Global Health Data Exchange, available at: http://ghdx.healthdata.org/) . GBD 201ta.org/) . The GBDMNDs are a set of chronic, degenerative, and progressive neurological conditions typified by the destruction of motor neurons and the subsequent deterioration of voluntary muscle activity. The most common MND is ALS. The International Statistical Classification of Diseases and Related Health Problems (ICD)-10 code corresponding to MNDs is G12. The GBD Study's gold standard diagnostic criteria are the El Escorial Criteria, combined with other similar criteria if necessary , 13, 14.*,\u201d and \u201cinciden*.\u201d had MND in 2019. The number of patients with MND in 2019 was 1.7 times higher than in 1990 . The age-standardized incidence of MND in 2019 was 0.79 , and the number of patients was 63,700 . The global age-standardized DALYs value of MND was 12.66 , and the count was 1034606.59 . The YLD and YLL values of MND were 57,068.01 and 977538.58 , respectively. The global death count of MND in 2019 was 39081.23 .High-income North America, Western Europe, Australasia, and Asia Pacific, as well as Southern Latin America had higher age-standardized prevalence rates. The age-standardized prevalence rates were low in the following regions: Oceania, Central sub-Saharan Africa, Western sub-Saharan Africa, Eastern sub-Saharan Africa, and Southeast Asia.The age-standardized incidence rates were high in Australasia, high-income North America, Western Europe, Southern Latin America, and high-income Asia Pacific and low in Southeast Asia, South Asia, Oceania, Andean Latin America, and Central Asia.The age-standardized DALY rates were high in Australasia, high-income North America, Western Europe, Southern Latin America, and Tropical Latin America. Central sub-Saharan Africa, Eastern sub-Saharan Africa, Western sub-Saharan Africa, Southern sub-Saharan Africa, and Central Asia had lower age-standardized DALY rates. The global age-standardized DALY rates of motor neuron diseases by age and sex are shown in The age-standardized rates of deaths caused by MND showed a similar pattern as DALYs. Australasia, high-income North America, Western Europe, Southern Latin America, and Tropical Latin America were the top five regions with high age-standardized death rates. Central sub-Saharan Africa, Eastern sub-Saharan Africa, Western sub-Saharan Africa, Southern sub-Saharan Africa, and Central Asia had relatively low age-standardized death rates.Changes in the age-standardized prevalence rates between 1990 and 2019 were most prominent in Australasia and Western Europe but lowest in Oceania and Central sub-Saharan Africa.Changes in the age-standardized DALYs and death rates between 1990 and 2019 showed a similar pattern. The highest increase in the DALY and death were observed in Southern Latin America and the Caribbean. The lowest changes in the DALY and death were observed in Oceania and East Asia.Prevalence, incidence, DALYs, YLDs, YLLs, and death due to MNDs by country in counts and age-standardized rates for both sexes for 2019 are listed in In 2019, the age-standardized incidence of MND was low in Malaysia, Seychelles, Indonesia, Maldives, and Philippines. In contrast, Ireland, Finland, Australia, United Kingdom, and Andorra had high age-standardized incidence of MND.Age-standardized DALYs and death were high in the following countries: Ireland, Australia, Andorra, New Zealand, and Finland. The age-standardized DALYs and death were low in the following countries: Somalia, Central African Republic, Burundi, and Democratic Republic of the Congo South.Between 1990 and 2019, the DALYs rates were increased to the greatest extent in Barbados, Costa Rica, and Uruguay. The DALYs rates decreased in the following countries: Slovenia, Guam, Bosnia and Herzegovina, and Republic of Korea. Portugal, Italy, Lithuania, and Costa Rica showed the highest increase in age-standardized prevalence and incidence rates over the examined period. Sudan showed low DALY but an increased death rate in 2019 compared to 1990.The age-standardized prevalence rate was relatively high in high-income North America, Western Europe, Australasia, and high-income Asia-Pacific regions with high SDI levels but was low in the sub-Saharan African region . The ageWe evaluated the burden of MNDs worldwide in 204 countries and territories from 1990 to 2019 using spatial Bayesian analyses. According to 2019 GBD estimates, age-standardized prevalence were 3.37 per 100,000 population and age-standardized incidence were 0.79 per 100,000 person-years for MND worldwide. In 2019, age-standardized DALY rate were 12.66 per 100,000 population and age-standardized death rate were 0.48 per 100,000 person-years associated with MND around the world. Global prevalence and deaths related to MNDs increased every year without significant changes in incidence. More than half of the prevalence and deaths due to MNDs occurred in three high-income regions . In general, the prevalence, incidence, and DALYs value of MND were high in regions with high SDI, except in high-income East Asia where these values were relatively low despite similar SDI. These findings might suggest that not only sociodemographic development but also the genetic background might be responsible for the MND burden. Compared with the previous 2016 GBD MND results , may be one possible reason for the high ALS incidence in North American and European populations \u201328. Howed Korea) .C9ORF72 mutations, is more common in regions of European ancestry and might be the cause of high disability and death and relatively low age-standardized DALY number and rate in middle and low SDI regions. The exception of reduced DALY in the high-income Asia-Pacific region might be partially due to different frequencies of ALS subtypes, different ratio of familial ALS and the coincidence of non-motor phenotypes compared to other subcontinents. Bulbar-onset ALS, which is well-known for its poor prognosis compared to limb-onset ALS, is more common in regions of European ancestry than in Asia . Similarnd death \u201328.The number of DALY and age-standardized rates were consistently higher in males than in females in all age groups between 1990 and 2019. Because the effects of sex on survival were not dominant, this male preponderance of DALY might explain by the difference in prevalence between the sexes. The male preponderance in MND, especially in limb-onset ALS, was consistent with previous reports , 9, 31. The age-standardized rate of DALYs of MND dramatically increased after age of 50, with a peak at 70\u201379 years followed by a rapid decline in both males and females. Given that ALS, which accounts for the largest proportion of MND, has a very short mean or median survival of 24\u201350 months from symptom onset, the changes in DALYs according to age in our results are consistent with the previous results showing the highest incidence between 70 and 74 years of age , 32. TheThis study has the same general limitations that inevitably occur in the design of GBD studies . First, In conclusion, the GBD of MND provides information on worldwide epidemiology, social influence, and risk factors of MNDs by using a standardized protocol. The global burden of MNDs is continuously increasing, especially in middle- and high-income areas. Because the number of epidemiology studies conducted in South and Central Asia, sub-Saharan Africa, and Latin America is small, and there is a high possibility that MNDs are underdiagnosed in the local system, the actual burden is expected to be higher than the presented results. In addition, the aging of the global population is expected to increase the share of the social burden of, for example, ALS, a neurodegenerative disease that mainly occurs in old age. The results of our analysis of the 2019 GBD 2019 Study may offer objective, recent information for resource allocation and healthcare planning related to MNDs at global and national levels.http://ghdx.healthdata.org/gbd-results-tool.Publicly available datasets were analyzed in this study. This data can be found at: the Institute for Health Metrics and Evaluation (IHME) Global Health Data Exchange (GHDx), Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. Written informed consent from the patients/participants or patients/participants' legal guardian/next of kin was not required to participate in this study in accordance with the national legislation and the institutional requirements.T-JS: concept and design, statistical analysis, administrative, technical, or material support, final approval of the version to be published, and had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis. JP, J-EK, and T-JS: analysis and/or interpretation of data, drafting of the manuscript, critical writing or revising the intellectual content. All authors contributed to the article and approved the submitted version.This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2021R1F1A1048113 to T-JS). The funders had no role in study design, data collection and analysis, the decision to publish, or preparation of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Cancer is the second leading cause of death among adults in the United States. Since the initiation of the \u201cWar on Cancer\u201d by Richard Nixon in 1971, advancements in cancer screening modalities and treatment approaches have resulted in marked improvements in cancer-related outcomes. However, not all groups have benefitted equally from advancements in the diagnosis and treatment of cancer. Indeed, we see persistent inequalities across the cancer care continuum, including prevention, screening, diagnosis, treatment, survivorship, and end-of-life care. Population groups at elevated risk for cancer inequalities include socioeconomically disadvantaged populations, underserved rural populations, racial/ethnic, and sexual and gender minorities. Against this backdrop, President Barak Obama signed into legislation the Cancer Moonshot initiative, which aimed to reduce cancer inequalities by making more therapies available to more patients and improving cancer prevention and early detection efforts. In recent years, emphasis has been placed on understanding and addressing the role of social determinants of health (SDOH) on cancer-related health disparities. A better understanding of SDOH can provide researchers and health professionals with effective strategies for reducing cancer-related health disparities and promoting cancer prevention and control.The Interplay Between Social Determinants of Health and Cancer Related Health Disparities,\u201d we solicited articles reviewing and addressing the role of social determinants of health in cancer-related health disparities to give readers an overview of updated health disparities information for their potential use in cancer research and practice. The goal of the Research Topic was to address cancer-related disparities with multilevel approaches, measurable outcomes, and effective solutions focused on innovative and effective strategies to understand the impact of SDOH or improve cancer-related outcomes in diverse populations.The World Health Organization's Commission on the Social Determinants of Health has defined SDOH as factors in which people are born, grow, work, live, and age, and the broader set of forces and systems shaping the conditions of daily life that influence health outcomes . An exteThis Research Topic collected 12 studies examining the relationship between SDOH and cancer prevention, treatment, survivorship, and disparities. The majority of research focused on the analyses of individual-level SDOH , followed by community-level , and interpersonal-level SDOH. Several studies incorporated multi-dimensional SDOH at the individual, interpersonal, and community levels in the analysis to better understand cancer-related outcomes and disparities. The findings from the collected articles are summarized as follows.Su et al. analyzed the Behavioral Risk Factor Surveillance System survey data. Results showed that lower family income was associated with cancer survivors' poor mental and physical health. Ishino et al. analyzed survivorship outcomes among colorectal cancer patients using SEER data. Older age at diagnosis, female, widowed, and non-Hispanic White with localized staging were associated with the lowest survivorship class.First, two studies examined the relationship of individual-level SDOH on health outcomes and survivorship among cancer survivors by analyzing national representative datasets. Hu et al. analyzing the Surveillance, Epidemiology, and End Results (SEER) data, found higher rates of refusing recommended surgery among Asian/Pacific Islander and African American patients than white patients. Furthermore, racial disparities in the receipt of timely lung cancer surgery were identified among Louisiana patients by Neroda et al.. Black patients were more likely to have delayed surgery than white patients. Other SDOH were also associated with receiving timely surgery.Second, two studies examined the influence of individual and community-level SDOH on receipt of cancer-related surgery and treatment. Li et al. found that access to care factors were associated with the uptake of low dose computed tomography (LDCT) among screening eligible patients in an academic medical center. Performing multilevel analysis using Swiss nationwide data, Jolidon et al. found that higher income and married women were significantly associated with higher mammography uptake. Beyond individual factors, Tsui et al. found that the health-related social needs assessment (HRSN) could be conducted at the health care organizational level to improve cancer screening utilization in the Chinese American population.Third, three studies discussed multilevel SDOH associated with cancer screening utilization. Zhou et al. analyzed the patterns and trends of disease burden and risk factors attributable to ovarian cancer across age, socio-demographic index, regions, and countries in terms of cancer risks. Tseng T-S. et al. indicated that increased cigarette smoking and other risky behaviors were found among African Americans eligible for LDCT screening during the COVID pandemic. Two articles discussed the barriers and facilitators for accessing smoking cession services and treatments in cancer prevention strategies. Tseng T.S. et al. found that geographical distance was a significant predictor of attendance of smoking cessation counseling classes. Shorter traveling distances were associated with more class attendance. Matthews et al. developed a model for developing and implementing smoking cessation interventions via patient health portals for low-income patients to facilitate patient linkage to receive free telephone-based smoking cessation counseling. Lastly, Gehlert et al. use the Critical Race Theory to address breast cancer disparities at the population level with an emphasis on social factors .Fourth, other studies focused on identifying the mechanisms by which SDOH contributes to cancer risks, cancer disparities, and the development of cancer prevention strategies. https://www.cdc.gov/socialdeterminants/index.htm) (https://www.whitehouse.gov/briefing-room/statements-releases/2022/02/02/fact-sheet-president-biden-reignites-cancer-moonshot-to-end-cancer-as-we-know-it/) have shown impacts on cancer-related health disparities in existing literature. The Centers for Disease Control and Prevention (CDC) have emphasized the SDOH and promoted SDOH data, research, tools for action, programs, and policy to improve health disparities (dex.htm) . Presidenow-it/) . Nationaesearch) . Nationaesearch) . Severalesearch) . Researcesearch) . To faciesearch) , 13.Despite advancements in cancer screening, diagnosis, and treatment, cancer inequalities remain a persistent public health concern in the United States and globally. The papers presented in this Research Topic contribute to the overall literature on cancer inequalities by describing relationships between SDOH and cancer-related outcomes. Further, the studies presented included novel findings regarding the influence of SDOH at multiple levels, which has implications for future research and intervention development. However, additional research is needed to understand the influence of SDOH on diverse communities experiencing cancer inequalities and to identify the pathways by which SDOH impact cancer outcomes to guide the development of strategies to eliminate or reduce cancer-related disparities.T-ST and C-CL contributed to the study conception and drafted the first manuscript. AM reviewed the manuscript and did the interpretation of the discussion. All authors contributed to editing and revising the manuscript critically and approved the final version of the article to be published.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Electronic cigarettes have been widely used all over the world. It is not clear what the advantages and disadvantages of a novelty in daily life are that is attracting increasing attention. Up to now, no bibliometric studies on e-cigarettes have been published in databases. Therefore, we are willing to explore directions and research hotspots in this emerging field by using bibliometrics to analyze research areas, publishing countries and institutions, high-output authors, and future trends of e-cigarettes in recent years. Compared with the traditional review, the bibliometric study can provide some information on core journals, articles, researchers, institutions, and countries concentrating on this topic to guide experimentation strategies and funding decisions.A bibliometric analysis was performed by CiteSpace and VOSviewer up to April 2022 in the core collection of Web of Science. HistCite, VOSviewer, CiteSpace, and the R-based Bibliometrix 4.1.0 packages were used to analyze literature information, including year, journal, country, institute, author, keywords, and co-cited references.Research related to e-cigarettes has proliferated since its inception around 2010. A total of 2,302 studies were published in 689 journals by our search method. Nicotine and tobacco research was the most published journal. The most prolific country was the United States, while the most influential institution was Virginia Commonwealth University. Eight of the top ten authors were from the United States. Oxidative stress, high school students, smoking cessation, delivery, behavioral economics, and exposure were the top topics.As an emerging social phenomenon, research on e-cigarettes has increased significantly over the past decade, particularly from 2015 to 2020. The top three core journals are Nicotine and Tobacco Research, the International Journal of Environmental Research, and Public Health. Eisenberg-Thomas had published numerous articles on e-cigarettes that had been co-cited in many papers. Oxidative stress, high school students, and smoking cessation are the top three areas of e-cigarette-related research, which were also important areas for further investigation. An E-cigarette is a rechargeable lithium-polymer battery-powered atomizer that heats the e-liquid (nicotine may or may not be added) in the atomizer oil chamber. E-cigarettes were invented around 2007, and their users have increased exponentially in recent years. Many transnational tobacco companies also began entering the e-cigarette marketplace around 2013. Initially, the original purpose of e-cigarettes was to help people quit smoking, but the convenience of e-cigarettes may have increased the smoking rate of minors and young adults. U.S. high school students' use of e-cigarettes rose 80% in 1 year (2017\u20132018) for the past 12 years (2010\u20132022) with the keyword e-cigarette. The search strategy: Topic = (electronic cigarette) OR (e-cigarette). The article language was set to English. The WoSCC database is a very well-known database in the medical community. Because it has a large amount of literature citation and citation data, the literature related to bibliometrics in recent years. Most of the literature related to bibliometrics has been done through this database.The literature included in this study for analysis included only articles, excluding conference abstracts and conference proceedings, and the number of papers, citations, titles, authors, institutions, countries, keywords, journals, years of publication, references, and other information were statistically analyzed for bibliometric analysis. Two reviewers (Shihui Hong and Can Feng) independently screened all the literature, annotated, and extracted data from the selected papers, and discussed the literature with disagreement.A total of four software and software packages were used in this study to perform the package for bibliometric analysis, including HistCite, VOSviewer, CiteSpace, and Bibliometrix 4.1.0 based on the R language.We have used the Bibliometrix package, a bibliometric analysis tool based on the R language, to analyze leading countries, such as Country Scientific Production and radar map. Visual cluster analysis and timeline visualization of cooperation are formed by VOSviewer, which can present collaboration and temporal trends between countries, institutions, and individuals in graphical images.The HistCite software provides statistics on the year of publication, country, institution, core journals, h_index and authors, global total citation score (TGCS), and local total citation score (TLCS) for all literature. Cluster analyses are all formed by VOSviewer as well as Scimago graphica.The Bibliometrix package was used to analyze the source dynamics of core Journals. The dual-map is a typical method of CiteSpace, which can likewise present literature by category, publication time, references, keywords for cluster analysis, biplot overlay analysis, etc.We have used CiteSpace to cluster co-cited References, present them in chronological order and reveal the most powerful citation bursts. CiteSpace and the Bibliometrix package were used to show the analysis of keywords of co-Cited References.R2 = 0.601). Based on a linear fit, the number of studies will reach ~600 in 2022. This period has been artificially divided into three stages based on annual production and growth rate: the inception stage (2010\u20132013), the growth stage (2014\u20132018), and the maturity stage (2019\u20132022). In the inception stage, the number of articles on e-cigarette-related cardiovascular disease was <20 per year. In 2010, Vansickel AR and Cobb CO, who started working on e-cigarettes, wrote the first article. They did not expose them to measurable amounts but found that they inhibited the assessment of nicotine/tobacco withdrawal symptoms .During the growth phase of e-cigarette research, <200 articles were published per year, but the average number of publications increased by about 30 per year, with an average annual growth rate of about 40%. In the maturity phase of the study, the number of articles published per year was >300, with an average annual increase of about 23 articles per year and an average annual growth rate of 17.06% . The higThe selected e-cigarette-related literature in this study had 59,945 citations, with an average of about 26 citations per article. The initial phase literature's TGCS and TLCS were low; from 2013 to 2015, the TGCS increased yearly. By about 2017, the TGCS values were relatively stable, representing that e-cigarette research had entered a relatively mature stage .n = 196; 6.73%), and the third was Italy (p = 0.274) . Corresp; 3.81%) . In addi; 3.81%) , indicat= 0.274) . The vis= 0.274) . Since 2= 0.274) .n = 108), followed by the University of Calif San Francisco in the USA (n = 86), the University of Southern Calif in the USA (n = 74), and the University of Oklahoma in the USA (n = 47). The largest TGCS was that of the University of Calif San Francisco (UCSF) in the USA (cited 5 103 times), followed by Virginia Commonwealth University and the University of Calif San Diego . Cooperation between the organizations was relatively strong. The UCSF-centered group closely collaborates with other institutions , Goniewicz Maciej L from Roswell Park Comprehens Cancer Center (38 articles published), and Krishnan-Sarin Suchitra from Yale University (31 papers published) . EisenbeThe articles related to e-cigarette research selected for this study were published in a total of 689 journals. The top 10 most published journals are shown in The citation and cited status of journals reflects the thematic distribution ; cited jThe more cited studies have focused on smoking cessation and smoking among adolescents. The most widely cited article to date is the one by Goniewicz ML on the significant reduction in population exposure to specific tobacco toxicants by replacing tobacco with e-cigarettes (cited 980 times) 12). Gr. Gr12). Co-cited references are those references that have been co-cited in other publications . In thisFinally, we carried out a reference explosion. Citation bursts are those references that are frequently studied in detail by scientists in each field over a given time interval . Figure We collected and analyzed a total of 3,144 keywords from the literature and obtained 8 clusters by cluster analysis of the papers published by the United States, the United Kingdom, and Italy, suggesting that these three countries have played an essential role in driving the development of e-cigarette research. The United States is the leader in e-cigarette research, with far more publications than any other country occupying the most significant number of articles, with Virginia Commonwealth University publishing the most of any institution, and their team in the Department of Psychology conducting a great deal of in-depth research and scientific collaboration in the field of e-cigarettes. China and the United States had the most partnerships among the countries that collaborated. This highlights the contemporary trend of scientific cooperation between large countries. These findings indicated that they paid more attention to e-cigarettes. Even though e-cigarettes were produced for smoking cessation, they also had side-effects.Eisenberg-Thomas is a leading scholar in e-cigarettes in the United States and has published extensively on e-cigarettes, covering the study of e-cigarette regulatory policy \u201319. MostAbout 1/3 of the literature on e-cigarettes had been published in the top 10 journals. Primarily the top 3 journals had published more than 100 papers. The top three journals were Nicotine & Tobacco Research, International Journal of Environmental Research and Public Health, and Addictive Behaviors. All three journals focus on the behavior and impact of electronic cigarettes. For example, a recent study published in Nicotine and Tobacco Research found that different nicotine concentrations and more decadent flavors may lead to higher e-cigarette use had smoking cessation as the primary research content, indicating that smoking cessation is still the principal value of e-cigarettes and a research hotspot in academia. Besides, adolescent smoking was also a significant concern in this field. Goniewicz found that among smokers who were unwilling to quit altogether, using e-cigarettes instead of traditional tobacco could reduce the harm caused by smoking . Grana pAs research evolves, several emerging research areas become subjects of interest to researchers. Burst detection is a method used to identify sudden increases in the frequency of keywords and references appearing in a certain period and can identify research hotspots and important research literature. References and keywords outbreaks show that some projects have seen the highest outbreak intensity in the last 3 years. At first, the electronic cigarette had been implied to be a suitable device for smoking cessation alternative , and alsKeyword clustering analysis and burst detection also reflect recent research hotspots . KeywordDaily electronic cigarette use is independently associated with increased HR and BP and myocThe effects of electronic cigarettes on hemodynamics could be the result of the heavy metals or other unknown ones which contain adverse effects on vascular function and hemodynamics.Notably, e-cigarette-associated pneumonia is likely to be a significant research trend in the future. It is necessary to find relevant cases, etiology, and solutions to avoid causing more harm to electronic cigarette users.To our knowledge, this is the first study to analyze e-cigarettes research using a bibliometric approach. In contrast to traditional reviews, bibliometric analysis allows for the analysis of evolving research priorities and trends from a point in time. It enables the identification of essential articles as well as researchers and institutions.Our study also has some limitations. First, literature published outside the WoSCC database is missed, resulting in research bias. Second, most of the results in this study were based on machine algorithms and were slightly deficient in manual generalization. Third, all the included e-cigarette studies were published in English. It is possible that e-cigarette-related literature published in other languages was overlooked.The number of papers on e-cigarettes has increased year by year. We used bibliometric methods to analyze research on e-cigarettes over the past decade, with the United States contributing the most; Virginia Commonwealth University had the most publications among institutions. The top three core journals were Nicotine and Tobacco Research, International Journal of Environmental Studies, and Public Health. Eisenberg-Thomas published the most articles. Oxidative stress, high school students, smoking cessation, and tetrahydrocannabinol are the knowledge base for e-cigarette-related research. As research progresses, we expect that e-cigarettes will serve the purpose of smoking cessation and radically reduce the health damage caused by cigarettes.The original contributions presented in the study are included in the article/WY and MF contributed to conception and design of the study. ZY organized the database. SH performed the statistical analysis. CF wrote the first draft of the manuscript. ZH, WW, and FW wrote sections of the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version.This work was supported by the National Natural Science Foundation of China (NSFC) grant , Science and Technology Commission of Shanghai Municipality (STCSM) Research Fund and Yueyang Hospital Research Projects (2018YJ05).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Dear Editor,Monkeypox, an unusual viral disease that affects both humans and animals, has long been prominent in portions of Central and West Africa; it was first found in humans in 1970. The symptoms caused by Monkeypox have been shown in In response to the global spread of monkeypox, Bangladesh has become the first country to put restrictions on sailors. Despite the fact that no cases have been confirmed in Bangladesh, the government has issued a health warning owing to the spread of the monkeypox virus in other areas of the world. Unless there is an emergency, all crew members are not authorized to utilize shore permits at Chittagong Port, and signed-off crew members are subject to health examinations. Crew changes, port entry, and developing vaccination restrictions remain major worldwide issues for mariners post-COVID-19. As the world prepares for the disease's spread, neighboring nations such as China and India have discussed strengthening travel restrictions .Although there hasn't been a confirmed case of monkeypox in Pakistan, the virus will almost certainly spread there. Given the burden the COVID-19 pandemic has had on Pakistan's already-struggling healthcare sector, several preventative measures must be implemented to halt the virus's eradication . If monkThe first suspected case of monkeypox in India is a 5-year-old boy from Ghaziabad, Uttar Pradesh. Although the monkeypox virus has not yet been found in India, a sample from a suspected monkeypox case in Ghaziabad has been submitted for testing [Due to a lack of knowledge about the current monkeypox virus epidemic, the World Health Organization (WHO) is collaborating with affected countries and others to improve disease surveillance in order to identify and assist people who may be affected, as well as to provide information on how to manage the disease. Until then, nations are advised to follow current WHO guidelines. Furthermore, COVID-19 and monkeypox can coexist despite being from distinct viral families. The pandemic COVID-19 virus can make the body more vulnerable, increasing the likelihood of mortality in a cohabitation setting. Because of the failing economies and bad quality of the healthcare system caused by COVID-19, low- and middle-income nations are more likely to face further suffering.The present study includes printed and published information; therefore, formal ethical clearance was not applicable for this study.N/A.All authors meet the inclusion criteria, and all authors read and approved the final version of the manuscript.1. Name of the registry: N/A.2. Unique Identifying number or registration ID: N/A.3. Hyperlink to your specific registration (must be publicly accessible and will be checked): N/A.Mohammad Mehedi HasanDepartment of Biochemistry and Molecular Biology, Faculty of Life Science, Mawlana BhashaniScience and Technology University, Tangail, 1902, Bangladesh. Email: mehedi.bmb.mbstu@gmail.com.N/A.The authors declare that there is no conflict of interest."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "It is known that biological risk factors and lifestyle behaviors are important determinants of dementia. However, there has been yet to be sufficient evidence to prove that community-level social capital is one of the determinants of dementia. This retrospective cohort study is a large, long-term, population-based study that investigated the association between community-level social trust and the risk of dementia in the Republic of Korea.Data came from the Korean National Health Insurance Service database. The community-level social trust values were determined by the Korean Community Health Survey. The study population consisted of 1,974,944 participants over 50 years of age and was followed up from 1 January 2012 to 31 December 2019 with a latent period of 5 years from 1 January 2012 to 31 December 2016. Cox proportional hazards regression was utilized to obtain the adjusted hazard ratios (aHRs) and 95% confidence intervals (CIs) for the risk of dementia according to social trust quintiles.Participants within the highest quintile of community-level social trust had a lower risk for overall dementia and Alzheimer's disease compared to those within the lowest quintile of community-level social trust. The alleviating trend association of high community-level social trust on dementia risk was maintained regardless of whether the participants had health examinations.Our findings suggest that higher community-level social trust is associated with a reduced risk of dementia. Community-level social trust is a crucial indicator of dementia and improving community-level social trust may lead to a lower risk of dementia. Dementia is a disease with high prevalence and high societal burden . In 2019Social capital can be broadly defined as a \u201ccollective asset in the form of shared norms, values, beliefs, trust, networks, social relations, and institutions that facilitate cooperation and collective action for mutual benefits\u201d . While tAmong the various cognitive dimensions of social capital, social trust has been recognized as an important factor on people's health outcomes. Subramanian et al. investigThere are other studies from the Republic of Korea which also investigated the association between community-level social trust and various health outcomes such as metabolic syndrome (MetS), cardiovascular disease (CVD), and mortality. In those studies, higher community-level social trust was associated with better health status \u201315. MeanThe study population was obtained from the Korean National Health Insurance Service (NHIS) database (NHIS-2021-1-755). NHIS is an institution that provides compulsory health insurance and corresponding services for all Korean citizens . For allStudy sample selection is presented in We obtained community-level social trust values based on the Korean Community Health Survey (KCHS) , coordinThe attending physician has a duty to put the International Classification of Diseases, Tenth Revision (ICD-10) codes for the primary cause of hospitalization upon hospital admission. Dementia was measured using both medications and diagnosis for dementia , Alzheimer's disease and vascular dementia . Since dPotential confounders including age in years (continuous), sex , categorical area of residence , categorical household income in first, second, third, and fourth quartiles, and Charlson comorbidity index (continuous) were adjusted. The household income was calculated from the insurance premium. The individual-level confounding variables such as depression diagnosis , diabetes diagnosis , or cardiovascular disease diagnosis before 1 January 2012 were adjusted additionally.As criteria for determining the qualities of the study population, the Chi-squared test was exploited for categorical variables, and the analysis of variance was used for continuous variables. For dementia risk in accordance with social trust quintiles, the adjusted hazard ratios (aHRs) and 95% confidence intervals (CIs) were calculated using multivariate Cox proportional hazards regression. The relationship between social trust and the risk of dementia was stratified according to age, sex, household income, Charlson comorbidity index, depression, cardiovascular disease, and diabetes. We also conducted analyses of the risk of dementia according to social trust considering participation in health examinations. Finally, we provided the aHRs per one interquartile range increase of the community-level social trust to show that the results align with the result from the adjusted hazard ratios using quintiles of the community-level social trust values.p-value of <0.05 on a two-sided basis. The data collection and statistical analysis were carried out using SAS 9.4 .We defined statistical significance as a p-values < 0.001). The number of participants who previously had depression, cardiovascular disease, or diabetes was also measured, and there were significant differences in the distribution of all confounders .p-value, 0.005) and Alzheimer's disease decreased.In this large, long-term, population-based retrospective cohort study, higher community-level social trust was related to a lower risk of dementia. Among the three categories of dementia\u2013overall dementia, Alzheimer's disease, and vascular dementia\u2013the risk of overall dementia and Alzheimer's disease was reduced with higher community-level social trust. Even after additional adjustments with risk factors such as comorbidities, depression, diabetes, and cardiovascular disease, the results were preserved and statistically significant. Based on our information, this is the first countrywide cohort study to show an association of community-level social trust with dementia risk for an extensive general population worldwide.Previously, other studies explored the association of various components of social capital, including social relationships, support, and community engagement with dementia. Kuiper et al. performeMoreover, current literature suggests that depression is one of the influential determinants of dementia , 25. TheRecently, a cross-sectional study from Japan discovered that community-level social capital, including social trust, is associated with dementia . AlthougVarious mechanisms have been proposed to explain the significant association of social capital, including community-level social trust, with dementia. First, social relationship engagement in districts with a high social trust may delay the incidence of dementia with cognitive stimulation and effective stress management by alleviating harmful stress nervous system responses . Second,Lancet Commission, like education level and air pollution .The datasets are not publicly available as they are protected by the Korean National Health Insurance Service. Anonymized versions can be made available upon institutional approval. Further information is available via The studies involving human participants were reviewed and approved by Seoul National University Hospital Institutional Review Board (IRB number: E-1806-076-951). Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.Study concept and design: JH, SJP, J-KL, HJJ, and SMP. Acquisition of data: J-KL and SMP. Analysis and interpretation of data and critical revision of the manuscript for important intellectual content: JH, SJP, J-KL, HJJ, JO, SC, SJ, KHK, JSS, and SMP. Drafting of the manuscript: JH, SJP, J-KL, and SMP. Statistical analysis: JH and SJP. All authors contributed to the article and approved the submitted version.This research was supported by the Korea Disease Control and Prevention Agency (Grant Number: 2021-11-017). SJP received a scholarship from the BK21 FOUR education program from the National Research Foundation of Korea (NRF).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Correction: Trials 23, 208 (2022)https://doi.org/10.1186/s13063-022-06548-7Following the publication of the original article , the autThe project has received a research grant from the Instituto de Salud Carlos III, of the Ministry of Economy and Competitiveness (Spain), awarded in the 2019 call within the Action of the Health Strategy 2013-2016, within the National Programme of Research oriented to the Challenges of Society, within the National Plan for Technical, Scientific and Innovation Research 2013-2016, with reference PI19/00736, co-financed with ERDF funds from the European Union .The original article has been corrected."} +{"text": "Objectives: Adolescence is considered a vital time to address healthy attitudes and values towards an effective transition to adulthood. The aim of this review was to analyse self-concept, self-perception, physical exercise, and lifestyle in the late adolescent population.Methods: Systematic review of studies assessing the results by the Rosenberg Self-Esteem Scale, the General Health Questionnaire, the Activity Questionnaire for Adolescents, and the Health Behaviour in School-aged Children questionnaires in late adolescents. The PRISMA recommendations were followed. The CASPe quality-check system was applied, excluding articles with a score <8.Results: 1589 studies were found, and 69 articles were selected. Adolescents with high self-concept and self-perception tend to be emotionally stable, sociable, and responsible. No significant differences were found regarding self-concept and self-perception between different countries, but there were differences between men and women. Physical activity and healthy diet improve self-concept and perception of body image.Conclusion: Self-concept and self-perception are associated with responsibility, stability, and mental strength. Most healthy behaviours during adolescence are followed during adulthood. Socio-cultural level of Health Science students is a differential factor for overweight and obesity. Satisfaction with one\u2019s life is related to a lower number of illnesses, increased happiness, and better emotional well-being , and theMany authors coincide that adolescence begins at puberty and ends with the complete development of the organism and the onset of adulthood \u20135. DurinSelf-concept, self-esteem, and self-perception are interrelated and condition the lifestyle and health-related habits of individuals, particularly of adolescents.Self-concept is understood as the set of feelings that the subject has about him/herself. In adolescents, it is important since it is an indicator of an adequate physical, cognitive, behavioural, affective, and social integration of the individual . It is rFinally, self-perception is important for understanding how individuals think, behave, and relate to others. It is understood that self-perception includes those internally conscious and organised concepts that the individual has about him/herself. It is related to a greater or lesser extent to age, low levels of schooling, income, race, marital status, smoking, physical activity, alcohol consumption, presence of chronic morbidity, and body mass index , 18.Overweight and obesity are among the most important problems in adolescence due to their psychological and social impact. The expansion of new technologies has influenced the prevalence of sedentary lifestyles, unbalanced and hypercaloric diets, and social changes . MoreoveHealth education approaches are decisive in adolescence . Thus, tLiving a healthy lifestyle has an impact on the prevention of cardiovascular diseases, which are the main cause of premature death in industrialised countries. The influence of lifestyle is also determinant to reduce the onset of pathologies such as type 2 diabetes mellitus, hypertension, dyslipidaemia, overweight, and obesity , 25. OthComparing the self-concept, self-esteem, self-perception, and physical exercise that young people have depending on their culture is fundamental to understand the actions for improvement that should be carried out on lifestyle. Knowledge on whether there are differences with respect to the environment and socio-cultural level is necessary to understand how it influences the individual. It also would facilitate the positive approach and development of the adolescent. In order to assess the self-concept, self-esteem, self-perception, physical activity, and lifestyle of adolescents, this research team previously conducted a systematic review titled \u201cQuestionnaires assessing adolescents\u2019 self-concept, self-perception, physical activity, and lifestyle: a systematic review\u201d . The aim1. To analyse the results provided by the following questionnaires: Rosenberg Self-Esteem Scale (RSES), General Health Questionnaire 12 (GHQ-12), Physical Activity Questionnaire for Adolescents (PAQ-A), and Health Behaviour in School-aged Children (HBSC), on the late adolescent population.2. To evaluate possible differences in self-concept, self-esteem, self-perception, physical exercise, and lifestyle of young people depending on their culture.A systematic review of the Scoping Review type was developed, following the recommendations of the Preferred Reporting Items for Systematic reviews and Meta-Analyses, also known as the PRISMA statement Figure . A searcThere is no prior record of the study protocol. However, any study was acceptable regardless of its design as long as it assessed self-concept, self-esteem, self-perception, physical activity, and lifestyle with the RSES, GHQ-12, PAQ-A, and HBSC instruments on late adolescents. Therefore, the study population was late adolescents. Articles in English, Portuguese, and Spanish were included. The exclusion criteria were: studies not exceeding a score of 8 in the Critical Appraisal Skills Programme Espa\u00f1ol (CASPe) ; publicaAll articles meeting the eligibility criteria were selected. The PICO method was used for the investigation question, considering as patient/population: teenagers and undergraduates; Intervention: questionnaires about lifestyle; Comparator: International late adolescents; Outcome: self-identity, introspection, workout, and way of living. The date of the last search is 07 March 2022.DECS and MeSH descriptors and Boolean operators were used . A preliA critical reading of the articles was carried out and they were assessed using the CASPe system. This was performed qualitatively by the researchers. Discrepancies were identified and extraction was agreed upon to produce the data list.To structure the data, all results compatible with the use of the RSES, GHQ-12, PAQ-A, and HBSC questionnaires on late adolescents were searched at international level. The results of the different items were compared on the basis of self-concept, self-esteem, self-perception, physical exercise, and lifestyle among young people with socio-cultural differences. During the critical reading of the articles, their internal structure, validity, and reliability were observed. The assessment of the degree of recommendation, as well as the analysis of the study population, the variables, the interventions used, and the results are described in The following questionnaires were used: RSES to assess self-concept; the GHQ-12 for the assessment of self-perception; the PAQ-A to measure physical exercise; and the HBSC study, where the lifestyle of adolescents is reflected. All articles with a moderate and high level were gathered using the GRADE system . All artA critical reading of the articles selected from the consulted databases was carried out. This systematic review was peer-reviewed by the researchers, deciding by consensus those issues that were not in agreement. The CASPe system wAfter the search, 1589 studies were identified. The total number of articles eliminated for being duplicates, non-retrievable publications, not meeting the study objectives, and/or not exceeding the CASPe score wiThese are described in This was assessed with the CASPe criticalThe Rosenberg Self-Esteem Scale (RSES) measures the overall self-esteem of the individual. It is a two-dimensional scale: positive self-esteem and negative self-esteem , 15. It As already mentioned, self-concept and self-esteem are linked. Thus, according to the study by Daniela-Calero A, et al. , having G\u00e1lvez-Casas A, et al. showed tThe General Health Questionnaire (GHQ) is a tool for measuring minor psychiatric morbidity in community, primary care, or medical-surgical outpatient settings. It assesses a person\u2019s self-perceived health over the past six months . The GHQA study in a hospital institution in the city of Medell\u00edn with the GHQ-12 surveyedVidera-Garc\u00eda A, et al. indicateInchley J, et al. state thSoria-Trujano R, et al. concludeThe Health Behaviour in School-aged Children (HBSC) measures young people\u2019s lifestyle habits and creates health promotion tools. It evaluates socio-demographic variables, diet, hours of sleep, risky consumption, oral hygiene, sexual behaviour, physical activity and sedentary behaviours, family context, leisure time, schooling, social environment, general health and psychological status, and socio-economic data. It is an initiative promoted by the WHO at the international level. Its latest version was published in 2020.Spotlight on adolescent health and well-being\u201d [The report \u201cl-being\u201d establisl-being\u201d , 70.Regarding nutritional habits, the majority of young people at international level do not follow the indications, which is detrimental to their healthy development. For this reason, the level of overweight and obesity has increased considerably, affecting 1 in 5 adolescents .The results of the study \u201cHealthy lifestyles in Nursing students of the Cooperative University of Colombia\u201d show thaDespite the cultural level of university students in Health Sciences and their extensive knowledge of healthy nutrition , they doSalas-Salvad\u00f3 J, et al. report tSoria-Trujano R, et al. compare In the research carried out by Hernando A, et al. boys havThe PAQ-A assesses the physical exercise of the person in the last 7\u00a0days. It consists of 9 questions that measure aspects of the physical exercise performed by the adolescent. It also provides information on whether the person has been ill. It is evaluated by means of a scale of 1 to 5 points that establishes a graduation of the level of physical activity carried out. It allows us to know at what time of the week the person is most active , 68, 69.According to the study by Rizo Baeza MM, et al. , overweiAccording to Mart\u00ednez-G\u00f3mez D, et al. , physicaRuiz-Ariza A, et al. showed that men are more attracted to physical activity than women .The level of physical activity in a sample of adolescents aged between 14 and 17\u00a0years was analysed in Colombia . StatistAccording to Rinc\u00f3n Herrera AD, et al. , ColombiSynthesis results: the results of the synthesis of the individual works are shown in Publication bias: all articles that did not obtain a score greater than or equal to 8 in the CASPe tool werSelf-assessment of health should be incorporated into health surveys on a regular basis, as self-perception of health is a good predictor of morbidity and mortality . Men tenAs results with a moderate-high degree of recommendation (GRADE) , it shouWith a moderate level of recommendation, it was found that men have a better physical self-concept and self-perception of health than women . The preHealth science students have knowledge and tools to improve their lifestyle; however, they do not apply this knowledge to their own benefit , 24, 72.Regarding cultural differences, the data provided by the RSES on self-esteem and self-concept do not report significant differences between adolescents from different countries. This concept is fundamental during the youth stage because it directly influences the psychological and physical state of the person \u201333. SomePhysical exercise, assessed by the PAQ-A questionnaire, is shown to be lower in both Colombian and Spanish women than in men , 70. HowIn conclusion, comparing the self-concept, self-esteem, self-perception, and physical exercise of young people depending on their culture is essential to understand the actions to be taken to improve lifestyles. Knowing whether there are differences with respect to the environment and socio-cultural level is key to understand whether it influences the individual. Understanding and promoting a healthy lifestyle in adolescents helps to target and implement interventions from a preventive point of view. It also facilitates positive adolescent approach and development, and it reduces economic costs, since the correct approach to young people avoids future complications such as chronic non-communicable diseases and hospital admissions for this reason.A healthy young and adult population is beneficial not only for the individual but also for their environment, which benefits from having a productive and healthy community. Improving self-concept increases the likelihood of being a more sociable, responsible, and emotionally stable person, which is linked to a greater self-esteem.This study is conditioned by the validity and reliability of the data provided in the selected scientific articles. It is also conditioned by the search for articles in English, Portuguese, and Spanish. It is influenced by the selection processes of studies where the CASPe and the"} +{"text": "Background: Hiring athletic trainers (ATs) in high schools has attracted rising interest as a potential way of improving adolescents' health by enhancing their safety and reducing their risk of injury.Objective: This study aims to determine if there is a difference in the referral patterns, injury diagnoses, and injury treatments performed at a metropolitan high school when an AT is employed versus not employed by the school.Design: This is a retrospective quantitative two-period study.Setting: The study was conducted in the high school athletic department in Norfolk, Virginia, and the study population was high school athletes (age 14-18).Main outcome measures: Changes in referral patterns, injury diagnoses, and injury treatments performed at a local high school when an AT is employed versus not employed by the school; specifically, we examined the number of and percent changes in yearly treatments, referrals, evaluations, and re-evaluations during the two periods.Results: Our first t-test revealed a statistically significant increase in the number of reported injuries between 2011-2015 and 2016-2020 and 2016-2020 , p = 0.014. The third t-test revealed a statistically significant increase in the number of treatment items between 2011-2015 and 2016-2020 , p = 0.01.Conclusions: The present study found an increased frequency of reported injuries, referrals, and treatment after ATs directly joined the staff of a large metropolitan high school. These findings suggest that direct employment of ATs is associated with greater recording of injuries and treatment of conditions. A reduction in referrals occurs with the presence of directly employed ATs, which could result in improved health for student-athletes, but this needs further study. Athletic trainers (ATs) are healthcare professionals who work in collaboration with physicians to provide a range of services, including injury and illness prevention, evaluation, diagnosis, treatment, and rehabilitation, as well as emergency care, to physically active individuals . Hiring Several associations, including the American Medical Association, the American Academy of Pediatrics Council on Sports Medicine and Fitness, the National Federation of State High School Associations, and the Appropriate Medical Care for Secondary School-Aged Athletes Task Force, have recommended that an AT be available to provide medical care for secondary school-aged athletes -7. DespiExternal ATs have limited access to student-athletes during the day compared to directly employed ATs, who have access to student-athletes all day. Furthermore, the availability of only one AT at a high school frequently means that an AT is unable to be physically present when multiple sporting events overlap. Such staffing shortages limit the AT's ability to provide basic but invaluable training and education to the coaching staff regarding basic emergency care . A recenBecause ATs possess a specific skill set and work in close collaboration with physicians, they play a vital role in coordinating and expediting care as deemed medically appropriate . In addiThe purpose of this study was to determine if there is a difference in the referral patterns, injury diagnoses, and injury treatments performed at a local high school when an AT is employed versus not employed by the school. We hypothesize that in direct employment.Study design\u00a0We conducted a retrospective review of de-identified archival data from the SportsWare\u2122 EMR electronic medical record used by the High School Athletics department, part of the Norfolk (VA) Public School system. No personally identifying data of any type was collected at any time. The procedure of the study was as follows: the total number of de-identified records and the aggregate number of evaluations, injuries, treatments, and referrals that occurred between 2011 and 2020 were collected from SportsWare. The pre-period for this study was 2011-2015, when the school district did not directly employ ATs, and the post-period was 2016-2020, when ATs were directly employed. A total of 2,177 athletes participated in the pre-period , and 2,109 athletes participated in the post-period . The same sports were offered at the high school in the pre and post-periods. Participants\u2019 demographic data for gender identity, ethnic identity, and age were not available. Data were compared between these two periods to determine if there were changes in practice patterns.Data collection\u00a0The data points we collected were the total number of injuries, the total number of initial evaluations by ATs, the total number of re-evaluations, the total number of treatment items, and the total number of referrals. The inclusion criteria for this study included high school athletes, aged 14-18, and evaluation, treatment, re-evaluation, or referrals by ATs during the study period. For this study, there was only one athletic trainer entering data per time period. We excluded data prior to 2011 and injuries that happened outside organized high school sports. We additionally excluded any instances of incomplete medical record documentation of an injury, treatment, or referral.Data analysisThe present study included one categorical level independent variable (time) with two levels: pre-period 2011-2015) and post-period (2016-2020). There were three dependent variables measured on continuous-level scales, including the number of reported injuries, the number of referrals, and the number of treatment items. The frequency of injury was defined as the number of injuries per year. The number of referrals was defined as any referral to an outside healthcare provider. Treatment items were defined as any rehabilitation service provided by the AT inside the high school. Statistical tests of mean comparisons for one categorical level independent variable with two levels (t-test) were performed using IBM SPSS version 26 with alpha set at .05 , t(3) = 6.82, p = 0.006, d = 5.61. During the pre-period (2011-2015), 13% (n = 295) reported an injury. During the post-period (2016-2020) 57% reported an injury. A second t-test was run to determine if there were differences in referrals between 2011-2015 and 2016-2020. Results revealed a statistically significant increase in the number of referrals between 2011-2015 and 2016-2020 , 95% CI , t(4) = 4.14, p = 0.014, d = 3.43. A final t-test was run to determine if there were differences in treatment items between 2011-2015 and 2016-2020. Results revealed a statistically significant increase in the number of treatment items between 2011-2015 and 2016-2020 , 95% CI , t(2) = 9.29, p = 0.011, d = 2.50.The findings of the present study demonstrated statistically significant increases in the frequency of reported injuries, referrals, and treatment items between the pre-test period 2011-2015) and the post-test period (2016-2020). The effect sizes of the results were all in the very strong range based on previously published guidelines [ and the The methodological limitations of the present study should be considered when interpreting the findings. We collected data from one high school located in the Norfolk County area; this comes with the caveat that our findings might not generalize to high schools in other locations. To address this, a larger study with more schools could be conducted. Moreover, the archival nature of our data collection method limited access only to demographic variables. Accordingly, we were not able to examine potential interaction effects between demographic variables and ATs across outcome variables . Future researchers can extend this line of inquiry by recruiting a larger sample and testing for multivariate interaction effects by demographic variables, for example, age, gender identity, ethnic identity, and sport.ATs provide a variety of services to the athletes they work with, including diagnosis, treatment, and evaluation of injuries. In support of this, numerous professional societies, including the American Medical Association, the American Academy of Pediatrics Council on Sports Medicine and Fitness, the National Federation of State High School Associations, and the Appropriate Medical Care for Secondary School-Aged Athletes Task Force, have recommended that an AT be available to provide medical care for secondary school-aged athletes -7. DespiOur study sought to determine if there is a difference in the referral patterns, injury diagnoses, and injury treatments performed at a local high school when an AT is employed versus not employed by the school. We found increases in the frequency of reported injuries, referrals, and treatment items. Although this study is limited to data from only one high school, the difference between the pre- and post-test groups was very large. Collectively, the results of this exploratory study suggest that the direct employment of ATs might have utility in promoting safety for high school athletes. Future research is necessary to continue to examine the work model of AT employment at high schools and other venues to quantify its value and identify areas for improved deployment of ATs."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "The recent 2022 monkeypox outbreak poses an emergency international public health concern amid rising cases as an all-time biggest outbreak . Monkeypviz., Congo Basin clade and the West African clade may also be useful to treat critical MPX cases such as in pregnant or breastfeeding women, pediatrics, or the immunocompromised \u20136. The uThe current 2022 MPX outbreak in many countries at once is atypical to previous instances. MPX reportedly spread among the MSM in the non-endemic regions through close contact like touching, kissing, and oral and penetrative vaginal or anal sex with infectious individuals . The rasWhen the WHO declared MPX a \u2018public health emergency of international concern' in June 2022, some 3,000 cases were identified in nearly 50 countries. More than 75,000 cases across 109 countries and territories are reported as on October 21, 2022 . Importa2, comprising mainland South-East Asia , and maritime South-East Asia . For various reasons, from a pandemic viewpoint, South-East Asia is a delicate and sensitive region. These reasons include its population density and the overall percentage of the global population, the extremely porous international borders for anthropogenic and economic activities, global reach being well-connected to the world by air, and the outward spread of the population .Recently, India reported discrete MPX cases from districts of Kerala and Delhi. So far, India has reported 14 MPX confirmed cases along with some suspected cases in a relatively short time. These are the first reported cases by the WHO in South-East Asia . To dateEarlier, 75 suspected MPX-related deaths were reported in Africa, mostly in Nigeria and the Congo, supposed to be more lethal MPX than in the West . As per As per a recent ICMR study, the retrieved MPXV sequences from India belong to A.2 lineage. The MPXV strains sequenced from Indian cases revealed that the virus had undergone significant genetic changes as noted by APOBEC3 gene mutation and an additional 16 SNPs . Thus, tA recent report indicated an alarming upsurge in diarrhea cases in Bangladesh which makes the case worse in this South-East Asian country amid the ongoing COVID-19 pandemic. About 4.5 million across the country are diarrhea infected in the first 3 months of the year . Healthchttps://www.livemint.com/news/india/monkeypox-8-cases-in-india-so-far-1-death-10-things-to-know-11659430193160.html). The COVID-19 mistake must not be repeated lest the world should pay the price for having remained inadequately responsive is set to manufacture a vaccine against it. Currently, the licensed vaccine for MPX is of a Danish firm, and the live virus on which it is based is stored at two locations, at the State Research Center of Virology and Biotechnology Institute, Koltsovo, Russia, and the Center for Disease Control, Atlanta, Georgia. The National Institute of Virology reportedly has been successful in isolating the MPX virus from a patient sample that shall help develop a vaccine and test kits (sponsive . Since ssponsive . Vaccinesponsive .RKM designed the study and made the first draft. MA, RNS, and AKS teamed up during the first draft. SM, VK, MK-E-Z, and NA updated the manuscript and edited it. KD and GP reviewed the final draft. All authors have critically reviewed and approved the final draft, and are responsible for the content and similarity index of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Objectives: We aimed to evaluate global epidemiological features of human monkeypox (mpox) cases and their associations with social-economic level and international travel arrivals.Methods: We estimated the pooled value by random-effects models. Then, we conducted an ecological study to evaluate the relationship of confirmed cases with social-economic indices and international travel arrivals using correlation analyses.Results: The average age and comorbidity rate of mpox cases in the 2022 human mpox outbreak were significantly higher than those of cases before 2022. During the 2022 mpox outbreak, the proportion of men who have sex with men (MSM) was high . The number of confirmed mpox cases in 2022 significantly correlated with high social-economic levels and international travel arrivals .Conclusion: Our findings highlighted the importance of early surveillance and timely detection in high-risk populations, including older people, MSM, and travelers, which is crucial to curb the wide transmission of mpox. As of 21 September 2022, human monkeypox (mpox) had spread across 106 countries or territories , 4. HumaHowever, at the beginning of May 2022, after the United Kingdom informed the World Health Organization (WHO) about a confirmed case of mpox who returned from Nigeria to the United Kingdom, there were subsequently clusters of mpox virus infections in multiple non-epidemic countries , 10. BetWith the ongoing 2022 multiple-country mpox outbreak, the WHO is calling for more research to understand the differences in mpox epidemiology from that before 2022 . One stuConsidering the above information, we need to explore the differences between cases before 2022 and those from 2022. Although one study has summarized case fatality rates (CFRs) of mpox before 2022, other epidemiological parameters, including incubation period, the secondary attack rate, animal contact history, and travel history, were not assessed . In addiIn our study, we extracted data from the literature on mpox cases, including suspected cases, confirmed cases, probable cases, and possible cases. A specific case definition is shown in Studies irrelevant to the subject of the meta-analysis, studies with insufficient data, duplicate studies or those with overlapping participants, modeling studies that did not provide original data, and non-human studies were all excluded. Full-text articles were then critically evaluated independently by two researchers (SMZ and HMS) to determine whether at least one of the review objectives was met. For the eligible articles, data extraction was done independently by two authors (SMZ and HMS) with any disagreements arbitrated by a third author (MD). In addition, we collected unpublished data from five sources, namely the websites of the WHO , United States Centers for Disease Control and Prevention (CDC) (29 June 2022), African CDC (14 June 2022), Nigerian CDC (14 June 2022), and ProMed (14 June 2022). One researcher performed search of the gray literature (MD), and two researchers (SMZ and HMS) reviewed the findings and added the relevant information to the data extraction sheet.Surveillance, case investigation, and contact tracing for monkeypox\u2014Interim guidance from the WHO [The included articles were case reports, epidemiological studies, and surveillance data from the websites. For these types, no formal checklists for critical appraisal are available, so informal quality assessments were performed. Information on study quality was added based on a self-reported assessment with a total score of 10 , p. 2. . This se the WHO .https://data.worldbank.org/indicator/SP.POP.TOTL) [https://ghdx.healthdata.org/record/ihme-data/gbd-2016-healthcare-access-and-quality-index-1990-2016) [https://hdr.undp.org/data-center/human-development-index#/indicies/HDI) [http://ghdx.healthdata.org/record/ihme-data/gbd-2019-socio-demographic-index-sdi-1950-2019) from World Bank and Global Burden of Disease Study (GBD) to explore the disparities in social-economic levels among different countries and territories during the 2022 multiple-country mpox outbreak. HAQ is calculated based on principal component analysis, providing an overall score of personal healthcare access and quality on a scale of 0\u2013100 by the GBD team [per capita [https://data.worldbank.org/indicator/ST.INT.ARVL?name_desc=false) [We extracted data on the total population (OP.TOTL) , healthc90-2016) , 24, humies/HDI) , and socGBD team . GBD resr capita . Additioc=false) . Internac=false) .In the meta-analysis, the specific calculation method for average values and standard deviations of each study was based on common and optimal estimate methods \u201331. BecaIn the ecological study, we presented the relationship between confirmed cases of mpox cases and HAQ (2015), HDI (2019), SDI (2019), and international travel arrivals (2019) using bubble and scatter charts. Moreover, their correlations were evaluated by Pearson correlation analyses.p < 0.05 indicated statistical significance.All of the data analyses were completed using R software version 4.0.5 (R Foundation) and Stata 16.0 . Two-sided The search strategy yielded a total of 2,864 publications, 180 of which were selected for full-text screening. Of these, 78 articles were suitable for data extraction , 35\u2013108.Specific studies for estimates of demographic, clinical, and epidemiological characteristics are shown in We observed that the average age of 167 mpox cases reported in 2022 was higher than that of 3,346 mpox cases reported before 2022 . The comFor different clades, the comorbidity rate (%) of 1,468 mpox cases infected with Clade I was higher than that of 450 mpox cases infected with Clade II . In addition, the proportion of animal contact history (%) of 351 mpox cases infected with Clade II was higher than that of 4,994 mpox cases infected with Clade I .p = 0.536), proportion of male patients , average duration of symptoms , secondary attack rate (SAR) , average incubation period , proportion of animal contact history , proportion of travel history , and proportion of MSM indicated that there was no publication bias, except for the estimation of the comorbidity rate and the CFR .Sensitivity analysis showed that all results were stable , HDI in 2019 , and SDI in 2019 in the American region among the 55 countries, and it remained significant in the European region and high-income region .To the best of our knowledge, this is the first study reviewing the latest global epidemiological and clinical characteristics of mpox cases from 1970 to 2022 and then reporting estimates by two periods and clades. We estimated demographic characteristics , clinical characteristics (duration of symptoms and comorbidity rate), and epidemiological characteristics . We also reviewed maternal and fetal outcomes among pregnant women.The average age of mpox cases was 21.05\u00a0years. Mpox cases reported in 2022 were older than those before 2022. Our study estimated that the proportion of male patients was 57.9%, and it was higher in the European region. Up to now, the 2022 Mpox Outbreak Global Map shows that the top three countries are the United Kingdom, Germany, and Spain\u2014all in the European region\u2014the main epidemic region . Perez DThe average duration of symptoms was 11.41\u00a0days, and it was lower in the high-income region and the Americas than in the low-income region and African region. The high-income region and the Americas generally have a higher quality of medical service . In addiMpox virus is transmitted between animals and humans, and from human to human . HoweverThe strengths of this review are that it included a broad search strategy on mpox worldwide, without time or language limits, which reduced selection bias. In addition, there was a thorough review of the gray literature for comprehensive data extraction. However, there were some limitations. First, mpox may occur in some countries where it could be unreported or undetected; therefore, due to data availability, our results may underestimate the real-world data. Second, since specific data on the clades were infrequently reported, we assigned clades based on the geographical spread described by the WHO. However, these may not be fully consistent with the reported cases, so our results may have information bias. Third, although there were articles presenting data on the transmission of mpox, many studies did not attribute cases to animal-to-human transmission or human-to-human transmission. Therefore, we could not analyze the changes over time, CFR, and SAR among different transmission routes. Finally, there was a lack of studies that reported the proportion of MSM before 2022, so we could not compare it in 2022 with that before 2022. In addition, the majority of cases before 2022 in Central and Western Africa were never published; although we included data from African CDC and Nigerian CDC, data were skewed toward the 2022 cases due to reporting bias.In conclusion, our study provided the estimation of the average age, proportion of male patients, average duration of symptoms, comorbidity rate, CFR, SAR, average incubation period, proportion of animal contact history, proportion of travel history, and proportion of MSM. We observed that the average age and comorbidity rate in 2022 were higher than those before 2022. Confirmed cases of the 2022 multiple-country mpox outbreak correlated with international total arrivals in 2020 among 55 countries. The multiple-country outbreak of mpox in 2022 highlights the importance of urgent response and global cooperation in coping with the transmission and impact of the disease. Except for providing information on the pooled estimates, our study also emphasized the demographic changes and the comorbidity rate in 2022, compared with before 2022. Additionally, we focused on the high 2022 proportion of MSM and the positive relationship between travelers and confirmed cases in 2022. To understand and explore the changing epidemiology of the mpox epidemic, increased surveillance and timely detection are crucial tools, especially in high-risk populations, including older people, MSM, and travelers."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Adolescence is a period characterized as transitional and as such, it is full of complications and conflicts. Research of Intra-Personal Conflicts in connection with Psycho-Emotional Well-being (PEW) comprising three kinds of indicators: personality, cognitive-evaluative and emotional represents new scientific approach. This approach provides the opportunity to define the role of PEW in Intra-Personal Conflicts: Motivation Value Conflict (MVC) and Self-Estimate Conflict (SEC).Our aim was to study the severity of MVC and SEC, the interrelationship of these types of conflicts, and their connection with various PEW components.237 high school students were surveyed. Tests of MVC, the Self-Estimate Scale (SE), and the Level of Aspiration Scale (LA) were applied to measure the conflicts. The Scale of Psychological Well-Being, the Scale of Life Satisfaction, and the Dominant Emotional States Test were employed to measure PEW.The study revealed a high prevalence of Intra-Personal Conflicts in the sample. The adolescents all had high levels of Intra-Personal Conflicts; changes were found in all three blocks of PEW. In the group with a high level of MVC, the levels of Environmental Mastery and Self-Acceptance were significantly lower. Having high level of SEC went along with decreases in most indicators of the personal and cognitive-evaluative components of PEW: decreasing of Cheerfulness, Active Attitude to Life Situation and Life Satisfaction; there were changes in emotional blockage, including decreases in Stability and Emotional Tone, and increases in Despondency, Tension, and Anxiety.The study found the prevalence of Intra-Personal Conflicts in the adolescents. We showed that the personality and cognitive-evaluative components of PEW played the role of conflict moderators, while the emotional components were manifested as intra-personal conflict. Over the last decade, the problem of Psychological Well-Being (PEW) has attracted more and more attention, not only from psychologists, sociologists, and educators, but also from politicians, economists, governmental authorities, and the whole international community. The interdisciplinary nature of this problem has been indicated by D.A. Two approaches can be considered the most traditional: the hedonistic and the eudaimonic. N. Bradburn and E. Diener are the founders of the hedonistic approach to research on subjective Well-Being . The proWe based our study on the concept of Psycho-Emotional Well-Being (PEW), which combines these approaches, using such notions as personality , cognitiThe relationship between Psychological Well-Being and Intra-Personal Conflicts and their role in personality development is also subject to debate. Some scientists considerIntra-Personal Conflict is viewed as a clash of opposing trends, interests, ideas, and aspirations within an individual. Researchers pay the most attention to Motivation-Value (MVC) and Self-Estimate conflicts (SEC). Motivation-Value Conflicts (MVC) are operationalized through the study of the ratio of values an individual considers vital and their availability , 2015. MSelf-Estimate Conflict (SEC) is identified as a conflict between an adolescent\u2019s level of Self-Esteem (SE) and Level of Aspiration (LA). Self-Esteem as a person\u2019s general evaluation of his or her value is expressed as either positive or negative Self-Orientation. Scientists point to the special role of Self-Esteem in adolescence, and its connection with Academic Performance and Mental Health . It has However, there is far less information on the links between MVC, Personality Development, and Well-Being. Moreover, there are indications of these conflicts\u2019 negative impact on adolescents\u2019 Socialization and Adaptation. . Some stAs our analysis showed, the relationship of MVC and SEC with Psychological Well-Being indicators still remains understudied.The following tests were administered: the Intra-Personal Conflicts study ; the DemThe study sample was comprised of 237 high school and gymnasium students, ages 15 to 18 . The study was conducted in regular class sessions in 2018. The parents were given an explanation of the purpose of the study, and informed consent was obtained from them.To reveal the severity of MVC and SEC as varieties of Intra-Personal Conflicts in adolescence and find out the relationship of these conflict types with each other and various PEW substructures.We proceeded from the following assumptions: 1) Various types of Intra-Personal Conflicts, in particular MVC and SEC, can be interrelated; 2) Pronounced Intra-Personal Conflicts may have two-way relationships with some levels of PEW; and 3) Different substructures of PEW can be interconnected with Intrapersonal Conflicts in different ways. We assumed that personal and cognitive-evaluative substructures can act as predictors of conflicts, either weakening or strengthening them. The emotional substructures of PEW act as consequences of conflicts.The data analysis showed that the sample\u2019s general level of Psychological Well-Being and Life Satisfaction corresponded to mean values, and in the given sample, reflected a generally favorable picture of Maturation. However, there was a high variability of indicators where the mean values did not reflect its individual characteristics.The comparative analysis of the frequency of conflict manifestations revealed that both types of conflict were prevalent, with high levels of SEC accounting for the greater number of respondents (52%), and that of MVC for 19.3%.Table 1).The mean value of the total MVC index in the sample was 34.9 points, \u03c3 = 15.24, which indicated an insignificant degree of Dissociation and a gap between the Values and their Availability. Draw your attention to the wide variability of the MVC index , which significantly exceeded the standard values obtained by the author of the method . All itsThe sample mean data indicated that Neutral Zones (65.5%) were expressed in the Internal Conflict framework; these zones are characterized by Values Coincidence and their Availability for Satisfaction. Internal Vacuums which means Redundancy of Availability in the Absence of its value prevailed in 15.19% of respondents. More than 19% percent (19.3%) of respondents had a pronounced Internal Conflict due to the Unavailability of Desired Values. Valid differences in Value/Availability ratio were revealed in 8 out of 12 life spheres (with significance level p ranging from .004 to .000).The most significant discrepancies between Values and their Availability were identified by such indices as Happy Family Life, Love, and Financially Secure Life, whereas a decrease in incentive motivation was noted in Beauty of Nature and Art, Active life, Creativity, and Cognition, which are characteristic of Internal Vacuum. The values of Health, Friends, Freedom, Challenging Work, and Self-Confidence were confined to the Neutral Zone.Table 2).Given the high variability of the indicators, a further analysis was carried out on groups with different levels of MVC severity. The group division was based on a Conflict Integral indicator, which is the sum of differences between a Value and its Availability in various spheres of life. An index equal to or exceeding 50 points meant the unavailability of significant values and indicated an individual\u2019s Motivation Disintegration, Deep Dissatisfaction, and Blockage of Basic Necessities. The two groups we distinguished were: Group 1, which consisted of adolescents with a low MVC level (144 respondents), and Group 2, which had medium or high MVC levels (94 respondents). Girls outnumbered boys in Group 2 (p = .005). These groups differed in their total MVC indices (p = .000) and in conflict structure , more Neutral Zones (p = .000), and fewer Internal Vacuums (p = .000), while in Group 2, the percentage of Internal Conflicts (p = .000) and Internal Vacuums (p = .000) was significantly higher, with fewer Neutral Zones (p = .000). These indicators suggest the Disintegration of Motivation, as a high level of conflict zones coexisted with poorly represented zones with Value-Availability concordance. There were more zones with a Low Level of Values and High Availability of them. This suggests that these adolescents have what they do not need and miss what is really meaningful to them.A comparison of the Values/Availability ratio in different spheres of life revealed a lower level of divergence of values Active Life (p = .000), Cognition (p = .000), Beauty of Nature and Art (p = .000), and Creativity (p = .000) and their availability in Group 2, along with reduced motivation (Mann-Whitney U-test). At the same time, Friends (p = .000), and Family (p = .000) were the most conflicted spheres. Thus, in the adolescents with pronounced MVC, there was a change in conflict structure and range of represented life spheres. It is also noteworthy that they had SEC in the area of Self-Confidence.The analysis of differences in PEW indicators between the groups revealed differences in personality indicators: in Group 2, the Level of Environmental Mastery (p = .016) and Self-Acceptance (p = .005) was lower, while the Level of Personal Growth pursuit was higher. The differences in parameters of dominant emotional states were negligible.The analysis of Self-Esteem and Level of Aspiration levels revealed that the sample fit within a framework of the average statistical norm , with high individual variability (from 0 to 100), and lower values among girls. Partial indicators of Self-Esteem fluctuated from 59.6 points to 72.6 points on different scales and indicated its mean level. The adolescents rated Intelligence and Character the highest and gave lower scores to their Peers\u2019 Authority, Self-Esteem, and Manual Skills. Scores on the Level of Aspiration demonstrated high values , which reflected the optimism of both the boys and girls about their capabilities. Like for Self-Esteem, the results showed very large individual variation . The highest Level of Aspiration was in Intelligence and Appearance; all other areas were evenly distributed.Table 3.Under our methodology, an Indicator of Conflict was considered to be a discrepancy between the adolescent\u2019s Level of Aspiration (LA) and Self-Esteem (SE) (less than 8 points or more than 22 points) . On thisThe analysis of the differences in PEW indices between the groups showed that they differed in Life Satisfaction (p = .008), Autonomy and Environmental Mastery (p = .000), and Positive Attitudes and Self-Acceptance (p = .002), as well as in the Total Indicator of Psychological Well-Being (p = .000). All these indicators were significantly lower in adolescents with a high SEC level. Some differences were found in dominant emotional states. Group 2 adolescents showed lower Activity (p = .004), lower Cheerfulness (p = .000), decreased Life Satisfaction (p = .005), and increased Tension (p = .002). Thus, the conflict of Self-Esteem affected all three constituents of PEW: personality, cognitive-evaluative, and emotional.In order to confirm the results obtained in the analysis of differences, we undertook correlation and regression analyses. Correlation analysis was carried out on the groups with a high level of conflict. Among MVC indicators, the Level of Conflict in Freedom was found to be the most interconnected with negative links of PEW indices with Life Satisfaction, Positive Relationships, Self-Acceptance, Total Level of Well-Being, Emotional Stability, and Satisfaction as a sustainable state (six connections at p .001 \u2013 .005). The Conflict in Creativity had four negative connections: Environmental Mastery, Self-Acceptance, Total Well-Being, and Tranquility (p = .001). Dissatisfaction with Family was directly related to Environmental Mastery and Total Level of Psychological Well-Being (p = .005). The total indicator of Internal Conflict formed two negative connections, one with Autonomy and one with positive Self-Image. The high value of the parameter Neutral Zones was directly related to positive Self-Image (p = .001); the parameter Internal Vacuums was interconnected with passive Life Attitude (p = .005). Therefore, various PEW indices were found to be interrelated with manifestations of MVC. Conflicts in Freedom and Creativity tended to be most integrated into the PEW framework.The analysis of SEC relationships revealed 21 negative connections with all PEW indices. Various indicators of SEC tended to be linked with Life Satisfaction, Self-Acceptance (eight connections at p .017 \u2013 .001), Environmental Mastery, Autonomy, Total Level of Psychological Well-Being (nine connections at p .022 \u2013 .000), and Life Goals (one connection) (p = .002). Relationships with Emotional states (three connections at p .028 \u2013 .000) indicated that an increase in SEC was accompanied by a decrease in Vigor, Emotional Stability, and Self-Acceptance. Indicators of Total SEC Index and Self-Confidence Conflict were most involved in the structure of connections with PEW.Regression analysis was also carried out in the group with a high level of conflicts. It is significant that in the group with SEC, in addition to the size of the discrepancy between the Level of Aspiration and Self-Esteem, indicators of MVC were found to be dependent variables, and in the group with high indicators of MVC, SEC variables were found.Table 4). SEC was described by the model as accounting for 31.9% of the variance. Total Level of Psychological Well-Being and Environmental Mastery were the predictors in this model. The Total SEC indicator was included as a dependent variable in the second model (with variance 13.9%), with the independent variables being Life Satisfaction and Autonomy . In the third model, Self-Acceptance , Environmental Mastery , and Positive Relations with Others predicted 19% of the variance on Neutral Zones. The independent variables of Self-Acceptance , Positive Relations with Others , and Environmental Mastery predicted 16% of the variance for the Total Value Conflict index in the fourth model.We created four models that re\" ected the relationship of PEW with various types of conflicts: two models showed the relationship of PEW with SEC, and two models, the relationship of MVC with PEW (Table 5). In the first model (variance 37%) Internal Conflicts of Family Values , Cognition , Freedom , Total MVC index and SEC in Skills predicted the dependent variable Cheerfulness vs. Despondency. In the second model, the dependent variable Relaxation vs. Tension (variance 23.6%) was predicted by Internal Conflicts in Health , Freedom , and Family . In the third model, the dependent variable Tranquility vs. Anxiety (variance 21.1%) was predicted by Internal Conflict in Beauty of Nature and Art , Creativity , and SEC in Self-Confidence . In the fourth model, the dependent variable Satisfaction vs. Dissatisfaction with Life (variance 10.3%) was predicted by the levels of Internal Conflicts in Freedom , and Family .At the next stage of the regression analysis, the dominant emotional states were included as dependent variables, while the indicators of Conflicts were the independent factors. Four models were obtained ; SEC in Intelligence and Skills had four negative connections with MVC through such indices as Friends, Family, Love, and Health .The results of our PEW analysis revealed an overall positive picture of the respondents\u2019 maturation over high individual variability, which was consistent with the data obtained from other samples . Our stuThe respondents with high levels of MVC were characterized by a change in MVC structure. In the framework of conflict, an increase in conflict zones was combined with a decrease in Neutral Zones and an increase in internal vacuum zones. This indicated Unavailability of significant values, an increase in Value /Availability concordance, and a decrease in a number of zones of Motivation Level of Development such as Creativity, Beauty of Nature and Art, Cognition, and Active Life. The respondents with a high level of SEC showed a decrease in the Total and in all indicators of this conflict.i.e., all MVC indices are involved in a SEC conflict.The analysis of differences in PEW indicators revealed that in the groups with a high level of MVC, Environmental Mastery and Self-Acceptance were significantly lower, but Desire for Personal Growth was higher, which may indicate a motivating role of MVC and is consistent with the results for adolCorrelation analysis confirmed that an increase in the severity of Internal Conflicts was correlated with an increase in the indicators of Personality and Cognitive-Evaluative PEW indices, and was also accompanied by the changes in the emotional part of PEW. These changes in the emotional component were manifested by a decrease in Emotional Stability, Activity, and an increase in Depression, Tension, and Anxiety, which correlates with the \u201cAnxiety Triad\u201d described in SEC . In our We used two strategies to conduct the regression analysis, In the first strategy, the Personality indicators and Cognitive-Evaluative components of PEW were taken as independent variables, whereas indicators of conflicts were taken as dependent variables. The results of this strategy application showed that MVC and SEC predictors were low indicators of Life Satisfaction, Autonomy, Environmental Mastery, and a decrease in Overall Assessment of Psychological Well-Being. In MVC conflict, Self-Acceptance, Environmental Mastery, Positive Attitudes were the factors that contributed to an increase in a number of Neutral Zones, thus reducing Conflict Tensions. In the second strategy, the independent variables were indicators of Conflicts, while the dependent ones were dominant Emotional States. It can be seen from the regression models that Family Dissatisfaction, Cognition, Beauty of Nature and Art, Freedom, and Health in MVC and SEC were linked to an increase in Confidence, while a decrease in Motivation worked as a predictor of Despondency, Tension, Anxiety, and Dissatisfaction.Therefore, the results showed the role of different PEW indices in the Intra-Personal Conflicts of adolescence. Personality and cognitive-evaluative components played the part of predictors and possible moderators of conflicts. This view is consistent with the understanding of PEW personality parameters as sustainable personality traits which reThect its psychological maturity as a result of ontogenesis, and act as moderators in difficult life situations . CognitiFrom our study we concluded that negative changes affected not only Anxiety, but also other emotional states; they were observed in both SEC and MVC. The internal relationship between the two above-mentioned Intra-Personal Conflicts was most clearly manifested in the connection between the severity of SEC in Intelligence, Authority, Confidence, Appearance, and the Total SEC index with MVC in Financial Well-Being, Friends, Family, and Love, which indicates the most significant modern adolescence values. The interrelationship of conflicts suggested that there were common predictors of Intra-Personal Conflicts, possibly accounting for personality and cognitive-evaluative PEW indices.This study highlighted the prevalence of Intra-Personal Conflicts in an adolescent sample. The respondents with high levels of intra-personal conflicts showed changes in all PEW indicators: personality, emotional, and cognitive-evaluative. Decreasing of personal indicators of PEW such as Environmental Mastery and Self-Acceptance is in correspondence with high level of MVC. A high level of SEC is characterized by decreasing of the most of PEW indicators: personal \u2014 decreasing of Cheerfulness and Active Attitude to Life Situation; cognitive-evaluative \u2014 decreasing of Life Satisfaction; emotional - decreasing of Stability, Emotional Tone, increasing of Despondency, Tension, Anxiety. The study showed that personality and cognitive-evaluative components of PEW played the part of conflict moderators, while emotional ones were manifested as effects of Intra-Personal Conflicts. A decline in Environmental Mastery and Self-Acceptance was accompanied by a high level of MVC. A high level of SEC was characterized by a decrease in the majority of PEW indicators. Adolescents with MVC were motivated by a desire for Personal Growth, whereas those manifesting SEC showed an absence of conflicts in Family, Friends, Freedom, Creativity, and Self-Confidence, which created the basis for the reliance on these spheres of life.The study\u2019s major limitation was the imbalance of the sample by gender with a predominance of girls and by having groups with a high level of Intra-Personal Conflicts. This can be partly explained by the literature on the greater severity of conflicts in adolescent girls . However"} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Optimal control of diabetes and relevant risk factors substantially reduce the risks of chronic complications and mortality. We investigated all-cause mortality rate and major causes of death between 2007 and 2018 in patients with diabetes in Taiwan. This study was conducted using data from Taiwan National Health Insurance Research Database. We selected patients with diabetes diagnosed between 2007 and 2017 (grouped according to the year of diabetes diagnosis 2007-2010 vs. 2011-2017). Information on mortality and causes of death by the end of 2018 was confirmed through linking to the National Death Registry. Standardized mortality rate (SMR) were calculated by weighting the World Health Organization (WHO) standard population (WHO 2000-2025). More than 2.7 million of patients with diabetes were analyzed and a total of 566121 deaths were identified. Overall, the SMR was 11.72 per 1000 person-years. Patients with diabetes diagnosed in 2011-2017 had a lower SMR (8.42 vs. 12.92 per 1000 person-years) than those diagnosed in 2007-2010. Similar finding were noted regarding the major causes of death . Compared with patients who were diagnosed in 2008-2010, those who were diagnosed in 2011-2014 and 2015-2018 had a higher 3-year survival rate after the diagnosis of diabetes. Patients who were diagnosed with diabetes after 2011 had a lower rate of all-cause mortality and major causes of death, compared with those who were diagnosed before 2010 in Taiwan. The continuous increase in the number of patients with diabetes \u20133 makes Optimal control of diabetes and relevant risk factors could substantially reduce the risks of chronic complications and mortality \u201310. SincIn this study, we aimed to update the rates of all-cause mortality and major causes of death in patients with diabetes in Taiwan. We investigated the mortality rates among patients diagnosed in different time periods to examine the differences in outcomes.This study was conducted using data from Taiwan National Health Insurance Research Database (NHIRD). Since the launch of the NHI program in 1995, more than 99% of the inhabitants in Taiwan are covered by the program which represents an important source for healthcare quality assessments and researches. De-identified data were released by the Health and Welfare Data Science Center for analyses and research use. This study was conducted in accordance with the Declaration of Helsinki. The study protocol was approved by the Research Ethics Committee of the National Health Research Institute prior to study procedures.th and 10th Revision, Clinical Modification. For ICD-9-CM, the codes for diabetes diagnosis were 250.x. For ICD-10-CM, the codes were E0-E14. Patients were excluded from the analyses if they had missing information on sex or date of birth. Patients were grouped according to the year of diabetes diagnosis (2007-2010 vs. 2011-2017). Information on mortality and causes of death by the end of 2018 was confirmed through linking to the National Death Registry. To avoid confounding effect of diabetes duration on risk of mortality, we also examined the survival rate annually after diabetes diagnosed in different periods (2008-2010 vs. 2011-2014 vs. 2015-2018).We selected patients with diabetes diagnosed between 2007 and 2017 for analyses. Patents were considered as having diabetes if they had 3 or more outpatient clinic visits or one hospital admission with the diagnosis of diabetes in 1 year . We examined the crude mortality rate (expressed as per 1000 person-years) for all-cause mortality and top 5 causes of death in the study population, and in subgroups by sex, age, and year of diabetes diagnosis. We calculated standardized mortality rate (SMR) (expressed as per 1000 person-years) by weighting the World Health Organization (WHO) standard population (WHO 2000-2025). The annual survival rates after diabetes diagnosis were compared among patients diagnosed in different periods (2008-2010 vs. 2011-2014 vs. 2015-2018).The major causes of death in patients with diabetes between 2007 and 2017 are shown in The survival rates after the diagnosis of diabetes according to year of diagnosis are shown in In this study, we investigated all-cause mortality rate and major causes of death between 2007 and 2018 in patients with diabetes in Taiwan. We demonstrated that patients with diabetes diagnosed in 2011-2017 had a lower all-cause mortality rate (SMR 8.42 vs. 12.92 per 1000 person-years) than those diagnosed in 2007-2010. The findings were consistent in men and in women. Similar results were noted regarding the major causes of death . Our results suggested an outcome improvement in diabetes patients during 2007-2018 in Taiwan.All-cause mortality rate in patients with diabetes has declined in the past decades , 18\u201320. https://dep.mohw.gov.tw/DOS/lp-5069-113.html ). The respective rates of our study population were 29.64 and 11.72 per 1000 person-years of cancer mortality in 2007-2010 and 2011-2017 was 7.26 and 5.66, respectively Table\u00a04.on-years , 40 haveon-years , 40. Theon-years or hearton-years , 43. HenThere are several limitations in this study. First, we did not have data on glycemia, blood pressure, and lipid profiles. These are important factors related to diabetes care and risk of mortality. Second, we did not have data on relevant medications, such as glucose- and blood pressure-lowering drugs, as well as statins. The use of some guidelines-recommended drugs (e.g. statins) may help reduce mortality risk in patients with diabetes , 24. UnfIn conclusion, we demonstrated an outcome improvement in patients with diabetes in Taiwan. Patients who were diagnosed with diabetes after 2011 had a lower rate of all-cause mortality and major causes of death, compared with those who were diagnosed before 2010. These promising findings may have implications for healthcare systems.https://cch@nhri.edu.tw.The datasets presented in this article are not readily available due to privacy/ethical restrictions. Requests to access the datasets should be directed to C-CH, The studies involving human participants were reviewed and approved by The Research Ethics Committee of the National Health Research Institute. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.J-SW, H-YO, Y-SY, and C-MH contributed to conception and design of the study. Y-LW, C-CH, and C-MH organized the database. Y-LW and C-CH performed the statistical analysis. J-SW and Y-LW wrote the first draft of the manuscript. H-YO, Y-SY, C-CH, and C-MH reviewed and edited the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version.This research was funded by the Diabetes Association of the Republic of China and the Taiwanese Association of Diabetes Educators . The sponsors had no role in the design, execution, interpretation, or writing of the study.We thank the Health and Welfare Data Science Center for providing data and Institute of Population Health Sciences, National Health Research Institutes for performing analysis.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Society\u2019s Assets and Liabilities as of 31 December 2021:January 1, 2021 to December 31, 2021. The Society continues to be in very good financial health. For 2021, the net revenue collected by the Society was $1,553,409.96, with the main sources of revenue being our two journals, Molecular Biology and Evolution and Genome Biology and Evolution .This report covers the 12 months from In a normal year, most of the Society\u2019s expenses are dedicated to travel for the in-person annual meeting. However, the annual meeting was again canceled in 2021 due to the COVID pandemic. The bulk of the Society\u2019s 2021 expenses of $449,705.09 were towards payouts to journal Editors-in-Chief, journal editing and marketing efforts, the organization and implementation of the 2021 virtual meeting, and Society prizes such as the mid-career award, lifetime contribution award, and graduate student excellence awards.MBE revenue reflects only income from Open Access fees as opposed to complete or partial revenue from a subscription model. However, the 2021 MBE revenue is inflated due to revenue carry-forward from 2020, resulting from a one-time shift from journal volume-based accounting to calendar-year-based accounting.This 2021 report is the first where 100% of Society\u2019s Revenues and Expenses for FY2020:"} +{"text": "Cancer-affected patients experience high distress due to various burdens. One way to expand psycho-oncological support is through digital interventions. This protocol describes the development and structure of a web-based psycho-oncological intervention, the Make It Training optimized. This intervention is currently evaluated in the Reduct trial, a multicenter randomized controlled trial.The Make It Training optimized was developed in six steps: A patient need and demand assessment, development and acceptability analysis of a prototype, the formation of a patient advisory council, the revision of the training, implementation into a web app, and the development of a motivation and evaluation plan.Through a process of establishing cancer-affected patients\u2019 needs, prototype testing, and patient involvement, the Make It Training optimized was developed by a multidisciplinary team and implemented in a web app. It consists of 16 interactive self-guided modules which can be completed within 16 weeks.Intervention protocols can increase transparency and increase the likelihood of developing effective web-based interventions. This protocol describes the process and results of developing a patient-oriented intervention. Future research should focus on the further personalization of web-based psycho-oncological interventions and the potential benefits of combining multiple psychotherapeutic approaches. A cancer diagnosis is often associated with multiple physical and psychosocial problems and significant distress, which can persist for years after completion of treatment , 2. At tNevertheless, some burdened patients do not receive psycho-oncological support due to various barriers, including not being screened for distress, limited resources and low accessibility, especially in rural areas \u20136. At thOne way to complement the existing psycho-oncological support for cancer patients is the adoption of digital approaches. Many patients report utilizing the internet to access health information and emotional support \u201311. DigiAlthough more qualitatively sound research is needed , 16, preAccording to the clinical guidelines for psycho-oncology, psychological interventions should be patient-oriented and deliver psychoeducation, relaxation techniques, and evidence-based skills training to help patients deal with cancer-related challenges . Most exMoreover, research indicates that these beneficial effects of mindfulness extend to web-delivered MBSR . ACT, onWith an interdisciplinary team of psycho-oncologists, psychotherapists, medical specialists for psychosomatic medicine, graphic designers, and health informatics scientists, and with the involvement of cancer patient representatives, we have developed the Make It Training optimized: A web-based psycho-oncological intervention based on a combination of CBT techniques as well as ACT and MBSR. The self-guided intervention is aimed at equipping cancer patients with skills to manage and deal with cancer-related challenges. It is a thereby assumed to reduce distress. More information about the ongoing effectiveness trial is available in the published study protocol . In thisThe Make It development involved six steps : (1) A nIn step 1, a comprehensive online survey was conducted to assess the needs and demands of cancer patients concerning psycho-oncological eHealth applications. The aim was to improve the acceptance and effectiveness of digital psycho-oncological interventions.In step 2, a web-based self-guided intervention, the Make It Training, was developed based on step the needs assessment of step 1. The intervention\u2019s benefits, design, and layout were evaluated in a first acceptance analysis and a longitudinal observational study. The aim was to see whether this first version of the Make It Training was a satisfactory psycho-oncological eHealth intervention and to identify potential future points of revision.In step 3, a council of four female patients and one male patient was formed. They are members of cancer support groups and were contacted by the psycho-oncological units of the university hospitals of Essen, T\u00fcbingen, and Erlangen. Two members were nominated by a German umbrella organization for cancer self-help. The primary role of the patient council was to advise the project team concerning further planning and optimization of the intervention and related studies. The patient council was consulted regularly during the intervention development regarding content, usability, and communication with the target group.In step 4, the content of the intervention was revised based on the previous steps. It was ensured that each module was structured consistently so that the use is intuitive and the design is user-friendly. An intervention structure was developed based on the selected methods and practical exercises. Previous evidence-based digital interventions were used as starting point.In step 5, the programming of the web app was planned together with the Department of Digitization and Healthcare of the Fraunhofer Institute for Software and Systems Engineering (ISST). The ISST is part of the Fraunhofer-Gesellschaft, an international research and technology organization. The programming and graphic design aspects were discussed at regular meetings. Furthermore, the intervention was continuously tested by different users on different operating systems to evaluate the intervention program\u2019s usability and identify points for improvement.In Step 6, a motivation and an evaluation plan were developed to enhance user adherence and assess the intervention\u2019s relevant effectiveness parameters. The evaluation plan is based on validated assessment instruments to evaluate different parameters such as efficacy, satisfaction, usability, and predictors of usage of the intervention. In addition, self-generated items were developed to measure the benefits of the intervention in detail. As this paper aims to describe the development of the digital intervention, the evaluation plan will only be addressed briefly here. More information regarding the ongoing evaluation can be found on the German Clinical Trial Register or in the published study protocol .The pre-study by Ringwald et al. aimed toThe second step was to develop a first version of the web-based program \u201cMake It Training\u201d (Mindfulness and skills-based distress reduction in oncology) based on the psycho-oncological content topics identified in step 1. The Make It Training and the data of its first pilot study were described in depth by Ringwald et al. . The firThe first study assessing user acceptance showed considerable acceptance and satisfaction by cancer-affected patients . The traThe task of the patient council was to accompany the study through regular meetings and communication with the study team. They advised the research team regarding (1) the optimization of the digital training and the study process, (2) a patient-friendly user experience, and (3) the dissemination of the project among patients and self-help groups. The council members were previously or still are affected by a cancer disease and are interested in science and supporting psycho-oncological care. The process of patient involvement was in line with the Involve guidelines issued by a work group of the British National Institute for Health Research .Constant exchange and meetings with the patient council and the research team have occurred. Due to the COVID-19 pandemic, these meetings were held online. An overview of the cooperation process is presented in In the following, the intervention is described according to the TIDieR checklist , see Sup. Based oVaadin Version 14 . The Fraunhofer ISST specializes in the research and development of software technology for digital and data-driven healthcare. Furthermore, the patient council was regularly asked for feedback following new development steps or updates on the intervention. Overall, it took around 4 months to implement the intervention. The web framework rsion 14 was usedgoDB 5.0 was usedvia the study website.1 The study website is the central contact point for interested patients and healthcare professionals. Information about the ongoing effectiveness trial Reduct . To assess the efficacy and usability of the intervention, we used patient-reported outcomes measures (PROMs) and patient-reported experience measures (PREMs) to evaluate the web app, which is described in-depth in the published study protocol .Cancer-affected patients often suffer from high distress , 2. So fIn six steps, we developed an evidence-based, patient-oriented intervention, the Make It Training optimized. In the first step, the needs and demands assessment revealed relevant psycho-oncological content topics and showed demand for eHealth applications in psycho-oncological care. The first version of the interactive, web-based Make It Training was developed in a second step. A first acceptance analysis showed high acceptance and provided feedback for further development. The third step was the implementation and involvement of the patient advisory council, which contributed to the further development and evaluation of the web-based intervention. Step 4 constituted the further refinement of the Make It Training, resulting in the Make It Training optimized. The result was a multifaceted training that addresses 16 topics relevant to cancer patients. In step 5, we implemented the web app of the intervention and the study website. And last, step 6 led to gamification and notification to motivate patients and enhance their adherence.To develop a web-based psychotherapeutic intervention, it is essential to involve cancer-affected patients from the beginning. In doing so, we created a web-based intervention that addresses the demands and needs of cancer-affected patients. Due to the constant exchange and feedback processes with the patient council, user difficulties were identified, and constructive solutions were found in a cooperative exchange. In addition, the dissemination of the intervention was improved, and it was possible to draw the attention of cancer-affected individuals to the intervention. Furthermore, successful cooperation in an interdisciplinary team with different expertise, i.e., psycho-oncologists, psychotherapists, medical specialists, graphic designers, and health informatics scientists, was central to developing an effective web-based psycho-oncological intervention. Only through this interdisciplinary cooperation was it possible to ensure that patients could optimally benefit from the intervention in the context of their disease management.Our intervention constitutes a self-guided intervention without therapist contact. It is designed to help cancer patients manage disease-related challenges autonomously. While not all cancer patients need or wish for in person psychological support, it cannot replace face-to-face interventions altogether. This especially applies to patients suffering from severe mental disorders or impairments who require more extensive support. In addition, while we strived to make the intervention as intuitively usable as possible, the use of intervention calls for a certain affinity with using web-based applications. One challenge of developing e-mental health interventions is to compete with existing commercial products in terms of available resources. This might influence the web design and usability of the intervention compared to commercial products in the e-mental health sector. Furthermore, critical readers might point out that combining different psychotherapeutic methods complicates identifying which therapeutic elements of the intervention were helpful. However, the three therapeutic techniques have a large evidence base for psychological support of cancer patients , 41, 42.With the help of this protocol, researchers and the general public can gain further insights into the process and structure of our web-based psych-oncological intervention. To develop effective e- mental health interventions, psychotherapy research must be combined with the state of the art of effective online applications to establish an evidence base for e-mental health applications. The pending results of our current randomized controlled effectiveness trial will show, whether our intervention is indeed effective and easily usable. Furthermore, qualitative research in the form of structured interviews with participants as well as moderation analyses will provide more insights into how the intervention was perceived by users. Future research should examine whether the intervention is effective for different age groups and tumor entities. Furthermore, dismantling studies could shed further light on the potential of this multimodal approach for cancer patients. Lastly, tools to further personalize e-mental health interventions, such as individually tailored feedback and content , 57, 58,The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by the Medical Faculty, University Hospital T\u00fcbingen, Eberhard Karls University; Medical Faculty, University of Duisburg-Essen, Essen, Germany; Medical Faculty, University Hospital Erlangen, Friedrich-Alexander-University Erlangen-N\u00fcrnberg; Medical Faculty, University Medical Center Leipzig, Leipzig, Germany; Medical Faculty, University Medical Center of the Johannes Gutenberg-University Mainz, Mainz, Germany; Medical Faculty, Albert-Ludwigs-Universit\u00e4t Freiburg, Freiburg Medical Center, Freiburg, Germany; Medical Faculty, Klinikum Rechts der Isar, School of Medicine, Technical University of Munich, M\u00fcnchen, Germany; Medical Faculty, Justus Liebig University Giessen, Giessen, Germany; and Medical Faculty, Philipps University Marburg, Marburg, Germany. The patients/participants provided their written informed consent to participate in this study.MT, AB, JG, and YE obtained the funding. SZ was a senior consultant within the project and provided supervision. JG, JH, AB, AW, CS, JK, JB, SS, and CJ developed and further adapted the intervention and prepared the content for the e-health application. SD and MP were responsible for implementing the intervention into a web app. JG and JH drafted the manuscript. AB, JK, and CS drafted the sections specifying the app development and patient council involvement. AM-T, JW, AW, AD, AS, and JK oversaw the recruitment at their study site and provided feedback during intervention development and contributed to this manuscript. All authors critically reviewed the final version of the manuscript and approved it and contributed to the article and approved the submitted version."} +{"text": "The era of ever-growing worldwide energy requirements demands the development of new methods of energy conversion, where the design of novel materials and the improvement of the efficiency of existing ones are of great importance. Transition-Metal-Based Compounds for Electrochemical Energy Conversion Processes is an open Special Issue of Materials, which welcomes original and novel research aimed at the most up-to-date research on this topic.d orbitals allow the formation of a variety of polyhedrons differing by coordination number, which translates into a multitude of available crystallographic systems. On the other hand, TMs exhibit a high capacity for charge compensation and co-occurrence in multiple oxidation states, which affects the tunable electronic structure and vulnerability to defect formation in both cationic and anionic sublattices, respectively. The latter, in turn, makes it possible to direct transport properties depending on the application. Lastly, TM-based compounds can be proceeded by means of a large variety of processes, allowing great potential for shape and surface engineering and tailorable morphologies, as well as various non-stoichiometric, multicomponent, and multiphase compositions.Transition-metal-based compounds, including intermetallic compositions, oxides, and chalcogenides, are characterized by several unique properties, mainly related to their crystal structure, electronic structure, and defect concentrations. The tendency of individual transition metals (TM) and the presence of arbitrarily occupied The versatility of these compounds is manifested in their widespread use in energy-conversion-oriented technologies. One of the best examples is TM-based oxides belonging to the perovskite group, usually characterized by excellent thermomechanical, transport, and catalytic properties . By skilIn addition to the intrinsic properties of transition-metal-based compounds resulting directly from their structure, these materials are also capable of assembling into heterogeneous systems, allowing the optimization of synergistic effects and thus the modification of their final properties. An excellent example of this is the oxide\u2013sulphide system, where the reciprocal arrangement of the valence and conduction bands causes an increase in the absorption band of the light prominence, as well as the tunneling of carriers and an increase in their lifetime . As a reThis Special Issue, entitled Transition-Metal-Based Compounds for Electrochemical Energy Conversion Processes, welcomes papers focused on transition-metal-based compounds for next-generation energy conversion devices, including sensors, photovoltaics, fuel cells, thermoelectrics, and catalysis. It is our pleasure to invite you to submit a manuscript for this Special Issue. Full papers, communications, and reviews are welcome."} +{"text": "Given growing clinical needs, in recent years Artificial Intelligence (AI) techniques have increasingly been used to define the best approaches for survival assessment and prediction in patients with brain tumors. Advances in computational resources, and the collection of (mainly) public databases, have promoted this rapid development. This narrative review of the current state-of-the-art aimed to survey current applications of AI in predicting survival in patients with brain tumors, with a focus on Magnetic Resonance Imaging (MRI). An extensive search was performed on PubMed and Google Scholar using a Boolean research query based on MeSH terms and restricting the search to the period between 2012 and 2022. Fifty studies were selected, mainly based on Machine Learning (ML), Deep Learning (DL), radiomics-based methods, and methods that exploit traditional imaging techniques for survival assessment. In addition, we focused on two distinct tasks related to survival assessment: the first on the classification of subjects into survival classes to stratify patients in distinct groups. The second focused on quantification, in days or months, of the individual survival interval. Our survey showed excellent state-of-the-art methods for the first, with accuracy up to \u223c98%. The latter task appears to be the most challenging, but state-of-the-art techniques showed promising results, albeit with limitations, with C-Index up to \u223c0.91. In conclusion, according to the specific task, the available computational methods perform differently, and the choice of the best one to use is non-univocal and dependent on many aspects. Unequivocally, the use of features derived from quantitative imaging has been shown to be advantageous for AI applications, including survival prediction. This evidence from the literature motivates further research in the field of AI-powered methods for survival prediction in patients with brain tumors, in particular, using the wealth of information provided by quantitative MRI techniques. Artificial intelligence (AI) is a branch of computer science that has been successfully applied to the analysis and extraction of meaningful features from medical images, with various clinical applications . In partBrain tumors are among the top ten causes of death from cancer ,7 and caPartially prompted by this unmet need, recent years have seen an increasing interest in applying AI techniques to MRI. Great emphasis has been placed on radiomics, a technique which aims to extract quantitative and reproducible features from images, including complex patterns that are often not visible to the human eye ,27. SpecA growing body of evidence suggests that radiomic analysis of MR images, can aid OS prediction, while also influencing patient management ,41. TherThis manuscript is organized as follows. AND brain AND (tumor OR tumour) AND AND (pediatric OR paediatric OR adults) AND (MRI OR \u201cmagnetic resonance\u201d)A literature review was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines . PubMed no full-text available;no AI application or non-pertinent application;conference proceedings;books or book chapters;non-English manuscripts.All studies evaluating AI and ML models for survival prediction in patients with brain tumors were included in this study. The initial search returned 1889 results , with a significant imbalance of results from Google Scholar. Following manual elimination of duplicates, titles were carefully screened to identify relevant papers. Any work that matched at least one of the following exclusion criteria was excluded:Review of the titles narrowed the results to 144 articles, 59 papers from PubMed and 85 from Google Scholar. Subsequently, review of the abstracts further narrowed the results to 88 articles, 35 papers from PubMed and 53 from Google Scholar. Despite review of titles and abstracts, not all the articles found on Google Scholar were relevant, moreover, some were not indexed in PubMed, and, considering the target audience and the push towards translational applications, we decided to include in this survey only indexed papers. Hence, after final revision, 50 papers were deemed eligible and included in this review.Several metrics can be used to evaluate a model, the most popular and well known are accuracy, sensitivity, specificity, and the area under the receiver operating characteristic curve (AUC), illustrating the diagnostic ability of a binary classifier system as its cut-off value varies. When estimating the goodness of a model that predicts survival, using the concordance index (C-Index) as an assessment metric may be useful. To account for the heterogeneity of the methods among the selected papers, C-Index and accuracy were used as comparison metrics for the SC and OS tasks respectively.Recent years have seen an increasing interest in using AI applications for survival prediction and risk stratification. A series of studies focused on defining paradigms that show the potential combination of clinical and computer-aided methods. These pioneering studies laid the theoretical foundation for subsequent research based on ML and DL methods. Most of these studies focused on glioma and, in particular, GBM. However, some results can be generalised across the wide spectrum of brain tumors.Zacharaki et al. showed hEmblem et al. showed tSome studies ,60 propoThese papers served as a prompt for the scientific community, and also demonstrated the potential utility of using additional information in tResearch gained momentum and, while still focusing on ML and radiomics, as DL became more accessible, an increasing number of studies explored its applications in medical imaging. Several clinical, functional, radiomic and morphological biomarkers were increasingly being included in the portfolio of information used to predict survival, and found to provide added value.Kim et al. emphasiz6-methylguanine-DNA-methyltransferase [MGMT] promoter methylation and isocitrate dehydrogenase 1 [IDH 1] mutation) to predict OS. The RSF model integrated with radiomic, clinical and genetic features showed the best performance (AUC = 0.74), when compared to models that only considered one type of feature. Peeken et al. [Several studies highlighted that integrating multi-modal imaging and radiomic phenotyping was beneficial for OS prediction ,56,65,66n et al. obtainedn et al. showed tn et al. showed pA few studies ,37 exploPreliminary results of DL applications were also beginning to emerge. Nie et al. developeBetween 2019 and 2020, the focus shifted away from ML to pivot on DL, hybrid techniques and CNNs, which have become one of the reference paradigms. Several studies suggested that DL-based survival prediction can outperform ML-based ones. In particular, non-linear DL methods may be useful in survival studies .Way et al. identifiA variety of authors have also employed hybrid techniques ,79,80,81P value < 0.001.Recent studies have alsNumerous studies ,58,81,82Several studies ,86,87,88More recently, we observed a consolidation in DL applications such as CNN and, often, hybrid methods consisting of ML and radiomics.Chen et al. hypothesHuang et al. developeAlthough slightly inferior in performance, the approach published by Preetha et al. , may havVarious authors focused on the combined use of ML and radiomic techniques for OS prediction in brain tumors ,93. ChatThe main limitations we identified in the reviewed papers are related to the improper use of techniques and algorithms (often not state-of-the-art), the use of limited or sub-optimal datasets and the use of retrospective cohorts. The combination of these factors had a major impact on the performance of the different methods.For the SC task, Single Layer Neural Network , SVM 6464, DecisWe presented a general overview of the current literature on AI applications in predicting survival in patients with brain tumors, based on MRI. The use of AI-based techniques, such as ML and DL, appears beneficial to predict survival. After evaluating several applications, we ranked the best applications based on the performance of the different algorithms for the two tasks of interest (OS and SC). Different approaches showed high performance, and the choice of the best one to use is non-univocal and subject to different variables. Unequivocally, the use of features derived from PWI and DWI/DTI were of significant relevance for both tasks. Indeed, the use of quantitative imaging is undoubtedly advantageous for AI applications. This is of particular relevance in an era when fully-quantitative MR imaging methods are becoming increasingly available and proven to be reproducible across different vendors .The use of semantic features in addition to clinical and radiomic features proved of significant relevance for the OS task, similarly to the use of features from clinicopathological information. The use of multiparametric MR images, compared to unimodal ones, also leads to significant improvements. ML methods appeared to perform better for this task. The best four algorithms have C-Index values in the range 0.79\u20130.91. The best algorithms were ML methods employing: radiomic, clinical and semantic features , PWI andIt is also worth noting that state-of-the-art methods and algorithms may not be those used in international competitions which may have intrinsic limitations. For instance, BraTS is an international challenge focused on tumor segmentation and not OS classification ; therefore the selected features were not necessarily optimised for OS. Most of the methods presented in this context, either based on a typical ML or DL architecture, extract and use significant features to achieve the best possible segmentation (the primary task) and often employ these features also for the secondary task (OS prediction). Hence, OS prediction is performed on the features that were chosen to obtain the best possible segmentation, without building a model focused on OS prediction itself. Therefore, those non-optimised models may obtain worse performance than state-of-the-art methods exclusively focused on survival prediction.In conclusion, depending on the specific task, different algorithms perform differently. In particular, ML methods, integrated with additional information, including clinical, radiomic, semantic and DWI/PWI information showed the best performances for OS prediction. Without the use of this additional information, DL methods would have performed better.Future studies should focus on developing ML/DL models by combining different data sources , which are correlated and may provide complementary information for impr"} +{"text": "Correction: BMC Public Health 19, 1607 (2019)https://doi.org/10.1186/s12889-019-7755-4YG and ZXL conceived and designed the study. YDY, CW, JXL, TTY, SH, HBX, and YYC participated in the acquisition of data. YG analyzed the data. HJ, LQL, and PQF gave advice on methodology. YG wrote the draft of the paper. CH contributed to revising the paper. All authors contributed to writing, reviewing or revising the paper and read and approved the final manuscript. ZXL is the guarantors of this work and has full access to all the data in the study and takes responsibility for its integrity and the accuracy of the data analysis. All authors read and approved the final manuscript.In the original publication the auth"} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Nature Biotechnology 10.1038/s41587-021-01188-9, published online 7 February 2022.Correction to: In the version of this article initially published, there was an error in the affiliation listed for Tabula Sapiens Consortium collaborator Albert Wu. The correct affiliation is Department of Ophthalmology, Stanford University School of Medicine, Stanford, CA, USA. The error has been corrected in the HTML and PDF versions of the article."} +{"text": "As the first bibliometric analysis of COVID-19 and immune responses, this study will provide a comprehensive overview of the latest research advances. We attempt to summarize the scientific productivity and cooperation across countries and institutions using the bibliometric methodology. Meanwhile, using clustering analysis of keywords, we revealed the evolution of research hotspots and predicted future research focuses, thereby providing valuable information for the follow-up studies.We selected publications on COVID-19 and immune response using our pre-designed search strategy. Web of Science was applied to screen the eligible publications for subsequent bibliometric analyses. GraphPad Prism 8.0, VOSviewer, and CiteSpace were applied to analyze the research trends and compared the contributions of countries, authors, institutions, and journals to the global publications in this field.Frontiers in Immunology published the most articles (178) related to COVID-19 and immune response. Alessandro Sette (31 publications) from the United States were the most productive and influential scholar in this field, whose publications with the most citation frequency . Furthermore, the development and evaluation of vaccines might become a hotspot in relevant scope.We identified 2,200 publications on COVID-19 and immune response published between December 1, 2019, and April 25, 2022, with a total of 3,154 citations. The United States (611), China (353), and Germany (209) ranked the top three in terms of the number of publications, accounting for 53.3% of the total articles. Among the top 15 institutions publishing articles in this area, four were from France, four were from the United States, and three were from China. The journal The United States makes the most indispensable contribution in this field in terms of publication numbers, total citations, and H-index. Although publications from China also take the lead regarding quality and quantity, their international cooperation and preclinical research need to be further strengthened. Regarding the citation frequency and the total number of published articles, the latest research progress might be tracked in the top-ranking journals in this field. By analyzing the chronological order of the appearance of retrieved keywords, we speculated that vaccine-related research might be the novel focus in this field. Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is currently in global pandemic since its first emergence in late December 2019. Following the statistic from the World Health Organization (WHO), as of April 25, 2022, the cumulative number of confirmed COVID-19 cases exceeds 511 million with more than 6 million deaths . UndoubtThe pathophysiological features of SARS-CoV-2 infection involve viral invasion, malfunction of the immune response, dysregulation of the renin-angiotensin-aldosterone system, endothelial cell injury, and microcirculation dysfunction , in whicvia bibliometric methodology.The bibliometric analysis represents a methodology combining mathematics and statistics, which have long been applied to analyze the literature in quantitative and qualitative over time to depict and predict the publication trend in a certain research field , 19. MeaBased on the Web of Science (WOS), the current study was carried out to analyze the status of research on COVID-19 and immune response using bibliometric analysis. As the first bibliometric analysis of COVID-19 and immune responses, this study will provide researchers with a comprehensive overview of the latest research advances. We attempt to summarize the scientific productivity and cooperation across countries and institutions using the bibliometric methodology. Meanwhile, using clustering analysis of keywords, we revealed the evolution of research hotspots and predicted future research focuses, thereby providing valuable information for the follow-up studies.Web of Science (WOS) represents a comprehensive, multidisciplinary database that provides information related to references of all publications, authors, authors' affiliated institutions, publishers, etc. With the powerful function and citation reports, it can quickly target high-impact studies, find the research direction concerned by the authorities at home and abroad, and reveal the development trend of the subject. It has become the most adopted database for bibliometric analysis \u201320.We chose Web of Science Core Collation (WoSCC) as our primary database for performing bibliometric analysis, for which we limited the type of publications to original articles and reviews in English. The searching timespan was from December 1, 2019, to April 25, 2022. All data were obtained and downloaded from a publicly available database and did not involve any ethical issues requiring approval.Due to the daily renewal of the database, data retrieval was carried out within a single day on April 25, 2022. The applied search strategy was as follows: TI = [macrophage OR neutrophil OR (NK cell) OR OR (myeloid derived suppressor cell) OR MDSC OR (innate lymphoid cell) OR ILC OR (dendritic cell) OR DC OR (T cell) OR (T lymphocyte) OR (B cell) OR (B lymphocyte) OR (plasma cell) OR (regulatory T cell) OR (Treg) OR (monocyte) OR (immunoglobulin) OR immunosuppression OR (immune dysfunction) OR (immune response)] AND TI = [(COVID 19) OR (2019 novel coronavirus) OR (coronavirus 2019) OR (coronavirus disease 2019) OR (2019-novel CoV) OR (2019 ncov) OR (COVID 2019) or COVID19 OR (corona virus 2019) OR (nCoV-2019) or nCoV2019 OR (nCoV 2019) OR (2019-ncov) or (COVID-19) OR (Severe acute respiratory syndrome coronavirus 2) OR (SARS-CoV-2)] AND LANGUAGE: (English) AND DOCUMENT TYPES: (Article OR Review). Detailed information regarding to literature screening were presented in Two reviewers (YX and P-yZ) collected detailed information from all incorporated publications, respectively, including country/region, keywords, published journals, citations, H-index, and so on. Thereafter, the extracted data were processed and analyzed using Microsoft Excel 2016 , VOSviewer , and GraphPad Prism 8.0 . The figures of results were integrated by Adobe Illustrator CS6 .The H-index means that a researcher has published H articles, and each of his articles has been cited at least H times. It is a well-established indicator to evaluate the quantity and quality of academic output from a certain scientific researcher or country/region. The impact factor (IF) was determined by inquiring about the latest version of Journal Citation Reports (JCR). Analysis of characteristics concerning published journals, authors, and countries/regions, the total number of citations, H-index, and a total number of publications were conducted by applying Web of Science. Moreover, GraphPad software was used to visualize the data. VOSviewer could automatically map and visualize the network of co-authorship, keywords co-occurrence, citation, bibliographic coupling, co-citation, and research hotspots. We chose full-counting as the counting method, and threshold settings for minimum numbers were described in detail in the results section. In the cluster analysis generated by VOSviewer, each dot represented a publication or keyword, and the size of each dot represented the frequency of co-occurrence. Lines connected relevant publications or keywords, the thickness of which indicated their linking strength. Moreover, VOSviewer could visualize the average appearing time of each keyword, which implied possible research trends in a certain area. In the clustering analysis of CiteSpace, the keyword co-occurrence network was composed of multiple clusters, and each one was assigned a label. The number of the label represented the size of the cluster. Co-citation referred to the relationship between two references that were simultaneously cited by other documents. The co-occurrence of keywords referred to the simultaneous occurrence of some keywords in all analyzed publications with a certain frequency.Overall, a total of 2,200 publications retrieved from Web of Science met our inclusion criteria and were subjected to subsequent analyses. The total citation frequency of all included articles was 47,681, with an average citation per article of 21.67 and an H-index of 94.Among all eligible articles, the United States , China , and Germany ranked the top three countries in terms of publication numbers. Publications from the United States had 16,890 citations , accounting for 35.4% of the total citations, with an H-index of 56. The citation frequency of China was 12,737 , with an H-index of 45. Germany ranked third in terms of publication numbers, total citations, and H-index, followed by Italy, England, and France .In the thermodynamic chart of The top 15 most influential institutions and journals in the field of COVID-19 and immune response were shown in Frontiers in Immunology was significantly higher than that of the other journals, with 178 records. Vaccines published 54 articles in this area, which ranked second. The number of publications on Scientific Reports tied for third place (45 articles). Additionally, the Journal of Medical Virology also had 461 citations for 26 articles on this topic. It is notable that articles published on Science Immunology , Immunity , and Nature Communications were ~1/8 to 1/4 of those published on Frontiers in Immunology, while their citation frequency per article was significantly higher than that of the Frontiers in Immunology.As for the top-ranking journals issuing publications on this topic , the numThe top five authors with the most publications are shown in Cell by Sette A had the second citation . The third most influential article was a retrospective review performed by Chen YW from China, in which they revealed the phenomenon of the decrease, functional exhaustion of T cells in COVID-19 patients and a negative correlation between T cell count and patient survival . It was tations) , 30, 32.To understand the main research focus of the 2,200 publications in detail, we applied the VOSviewer Software to analyze the keywords extracted from the titles and abstracts, which were defined as words appearing at least 85 times. A total of 120 keywords met the requirements and were subsequently categorized into three clusters: clinical research, innate immunity-related research, and acquired immunity-related research . Within + T cell, and safety.As summarized in A total of 139 references cited at least 50 times were retrieved and eligible for analyzing co-citation, which was further classified into three clusters As shown in the density visualization of The United States, China, and Germany were the top three countries for the total number of publications, citations, and H-index among all countries/regions . China fAs depicted in The international cooperation of the top 15 countries/regions was shown in Frontiers in Immunology published the most articles with the highest total citations on COVID-19 and immune response compared to the others since this journal was open access with an irregular publishing period. Meanwhile, Frontiers in Immunology set up various sections related to COVID-19 by inviting professional researchers worldwide, leading to the publication of more influential and advanced studies. Notably, other journals with huge academic impact included Science Immunology, Immunity, Nature Communications, and Journal of Medical Virology also published several articles on this topic. Although few of their publications had relatively higher citation frequency per article, indicating their profound academic influence, we forecast that the latest research progress in this field may still appear in Frontiers in Immunology and those journals, as supported by their fame and impact in this area.In terms of institutions publishing research on COVID-19 and immune response, the University of California System in the USA ranked first, followed by Udice French Research Universities and the National Institute of Health and Medical Research in France. Institutions from France, the United States, and China dominated this research field, as evidenced by 4 of the top 15 institutions being from France, 4 of them from the United States, and 3 from China. As for the journals, As for the most prolific and influential authors in the field of immune response and COVID-19, the top three authors, Alessandro Sette, Daniela Weiskopf, and Alba Grifoni, were all from the team of the La Jolla Institute for Immunology in the United States. They have 21 co-authored articles. And research conducted by this team comprehensively covered the role of T cells in SARS-CoV-2 infection, clinical symptoms of COVID-19, and immunization vaccine. However, the co-authorship network in Clinical Infectious Diseases by Tian et al. mapping of T cell specificities might provide valuable targets for most vaccine development. In addition, the team determined the SARS-CoV-2-reactive CD4+ T cells in ~40\u201360% of unexposed healthy controls, implicating the common coronavirus had the identical cross-reactive T-cell recognition with SARS-CoV-2. Within similar items, Bertoletti et al. reported their investigations concerning cross-reactive T-cell recognition of SARS-CoV-2 in Nature and Tem (effector memory T cell) . Given tfunction . The trefunction . Furtherfunction , 40. TheThe clustering analyses of keywords reveal that vaccine development and efficacy evaluation will soon become a hot research topic in the future. Accordingly, there are 232 vaccine candidates in disparate stages of development worldwide, of which nine have been approved and put into clinical use in many countries . CurrentSeveral limitations should be considered when interpreting our findings. Firstly, a fraction of non-English publications was not considered due to our inclusion criteria. Omission of articles published in non-English language might inevitably render some inaccuracy. Secondly, we solely extracted records from the WOS, rather than databases such as Scopus and PubMed. Finally, since articles on COVID-19 and immunity were mainly published in the past 2 years and updated rapidly, we were unable to fit the growth curve for each country/region to precisely predict the quantity of publication in the upcoming years.Frontiers in Immunology, Science Immunology, Immunity, Nature Communications, and Journal of Medical Virology, will publish more influential work and elicit the latest progress in COVID-19 immunity. Alessandro Sette represents the most influential scholars on this topic. By analyzing keywords, the development and evaluation of vaccines become a novel research hotspot in relevant area.Taken together, the current study has summarized and elucidated the global publication trend of research on COVID-19 and immune response. Among all eligible records, the United States has the most publications with the highest H-index and citation frequency. China ranks second in the publication number, total citations, and H-index. Based on our bibliometric analysis, China still needs to strengthen its international cooperation and basic research. Scientific journals, including The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/X-mS, Y-mY, and R-qY conceived the analysis. YX and P-yZ extracted all data. Z-bT undertook and refined the inclusion process. YX, R-qY, and Y-mY co-wrote the article. R-qY, YX, P-yZ, L-yZ, and H-tZ undertook the bibliometric analyses. All authors contributed to and revised the final manuscript.This work was supported by grants from the National Natural Science Foundation of China , the Funds for Young Reserve Talents of Hubei Province of China (No. HBRC20200411), and National Tutorial System Training Program of Suzhou (2020-12).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Background: Preventive and health-promoting policies can guide (place- and space-specific) factors influencing human health, such as the physical and social environment. Required is data that can lead to a more nuanced decision-making process and identify both existing and future challenges. Along with the rise of new technologies, and thus the multiple opportunities to use and process data, new options have emerged to measure and monitor factors that affect health. Thus, in recent years, several gateways for open data have become available. At present, an increasing number of research institutions as well as (state and private) companies and citizens\u2019 initiatives are providing data. However, there is a lack of overviews covering the range of such offerings regarding health. In particular, for geographically differentiated analyses, there are challenges related to data availability at different spatial levels and the growing number of data providers.Objectives: This paper aims to provide an overview of open data resources available in the context of space and health to date. It also describes the technical and legal conditions for using open data.Results: An up-to-date summary of results including information on relevant data access and terms of use is provided along with a web visualization. All data is available for further use under an open license. Open Data (OD) made available by public authorities, private or state-owned companies, research institutions or citizens\u2019 initiatives allows novel approaches and options for use. The term Open Government Data (OGD) is currently used to describe a growing number of supply channels for administrative data, along with standards for research and provision, and the offering of selected data through company websites and/or general portals. Compared to other countries, however, Germany is still lagging behind when it comes to exploiting this potential for public health research , , , 10], , availabllegal openness. Most public data can only be used in accordance with declared conditions, e.g. allowing commercial use, processing and dissemination. For personal data protected by law, subsequent use is not permitted in most cases. Table 1 Technical openness relates to data formats ensuring reuse with both proprietary and free software. In recent years, recommendations for the publication of data have been formulated. The OGD principles listed in Table 2 Published data is not necessarily open. It is not open, for example, if rights of use are retained or granted on a case-by-case basis only, or if the data is provided in a format preventing further use. By definition, data is open in particular if it is machine-readable and structured and can be freely used, reused and distributed by anyone using an open interface for any purpose and without any restrictions, discrimination or fees . EfficieIn addition to OGD, open data that can be used for spatial public health research includes a wide range of open geospatial data as well as data from official statistics, which in some cases are already linked to geospatial data. Furthermore, companies, research institutions and the civil community offer data in open formats for further use. The sources shown in Figure 1 whether or as how open OGD can be described an anOpenStute data . It is nute data . By intrINKAR database of the Federal Institute for Research on Building, Urban Affairs and Spatial Development (BBSR). Social welfare statistics and social reporting also contain relevant OD at federal level and, in some cases, at the small-scale level, available at state or local government gateways. So far, however, such \u201catlases of social infrastructure\u201d have mainly been offered in non-machine-readable PDF format.Data from official statistics include geospatial as well as registry, social, environmental and health data. At federal level, the Federal Statistical Office is the leading data provider. It is also possible to obtain official OD from regional databases, local or municipal governments as well as from overarching portals and databases, such as the Deutsche Bahn AG is an example of a private company that offers OD on a national scale via its own gateway. Likewise, the Open Data Platform \u00d6PNV initiative, powered by nine major German transport associations, makes OD freely available on its portal. In addition, more and more regional and local transportation services are offering open data on their corporate websites (e.g. NVBW Open Data).The data portal of https://sensor.community/de/ and OpenSenseMap.org platforms as well as some projects of the Codefor-Lab (https://www.codefor.de/projekte/). The offenedaten.de portal, which has, however, been closed in the meantime and on which Germany-wide OD was mapped interactively for the first time on the basis of web-crawler matches, still serves as a prototype for such applications today , , and envienvironment and health at national and municipal levels, e.g. from environmental reporting (UBE) and the federal/state indicators of health monitoring, they do not refer to each other , , , with soFurthermore, the economic environment, the financial situation of the public sector, and the level of collective wealth reflect the social capital and the capacity of a region to provide services of public interest. A valuable synopsis of data resources along this dimension, including official statistics, welfare statistics and the GBE, has been elaborated by Bardehle et al. . InformaWell-being can also be affected by personal feelings of vulnerability (e.g. due to crime and traffic safety). Although the Federal Criminal Police Office (BKA) stopped providing its federal-level police crime statistics (PKS) exclusively as PDF files in 2014, small-scale data in machine-readable form is often missing or can only be accessed upon request.Access to (care) services relates, among other things, to both general availability and access in the sense of reachability. Social conditions and thus existing social infrastructures are of great importance to health due to their salutogenic protective function .Life expectancy at birth can be interpreted as an indicator of the general living and healthcare conditions in a given region. While family physicians represent a key institution in outpatient healthcare, recent years have witnessed an increasing need for certain healthcare specialists . In addiThe physical environment has a major impact on health, particularly on a small-scale level. This refers to the mental, physical and social components . The lanEnvironmental hazards are physical, chemical and biological factors affecting individuals externally. Of these, air pollution is considered to contribute to diseases with the greatest attributable burden of disease . CommonlHowever, with proximity to traffic being a relevant criterion, total annual emissions (in metric tons) in predefined administrative units are of restricted explanatory value. Thus, there is limited applicability for deriving area-wide, small-scale exposures (e.g. daily concentrations for fine particulate matter). This also applies to other data collected only at specific measuring points, such as meteorological data of the DWD. Direct environmental influences also appear as health consequences due to climate or environmental noise. Noise maps for road traffic are usually available for large cities with more than 100,000 inhabitants, in some cases as comprehensive model calculations and with data from noise appraisals. For smaller cities and communities, however, the situation is more varied. Likewise, climate data (and climate function maps) are available in some cases, especially for small-scale areas. Soil and water quality data is largely available at most scales.Although there is political and public interest in implementing health-promoting policies, especially on a municipal level, problems arise especially when it comes to small-scale analyses. For data-protection reasons, raw data is not provided for the smallest geographical units but processed or aggregated for specific purposes instead.Gov.Data can only host municipal datasets if the associated federal states (FS) support the portal. To date, however, only 12 FS have joined the agreement. Instead, some federal states have launched their own data portals. As a result, the amount of data, access to it and the legal framework are heterogeneous and, in some cases, do not come with any legal regulations based on the Freedom of Information Act (IFG). Even the municipal GBE is focused on a state- or region-specific perspective and does not exhibit a common basic standard.Moreover, different administrative boundaries and aggregations make it difficult to compare and link data with attribute data. Besides said limited availability and reduced ability for regionalization, data usage is mainly hampered by federal structures. For instance, central portals such as Despite the slight increase in the number of municipalities publishing OD , the provision is currently still very selective or, in some regions, almost non-existent Figure 2 . At diffhealth in all policies. Therefore, public health research seeks to make \u2018data for action\u2019 more visible, accessible and disseminable [Availability, interdisciplinary use and linkage of data are essential for the development and monitoring of health strategies and are in line with the understanding of eminable to identIn the light of ongoing digitization and the resulting increase in data quantity, quality and use, extensive opportunities have opened up for analyzing associations between space and health, ranging from regional to international levels.In this context, open access to data is a necessary requirement. The data resources highlighted above demonstrate that much data, in particular open geodata, is available in principle, but also that information on how to access it is crucial. Furthermore, in many cases it remains unclear what data is actually available as OD. It is difficult to determine the current supply status of OD, as there is no central or official body to collect this information.Due to some limitations, the increasing number of different portals provides, at best, an entry point for research. Freedom of information and data-protection laws, which are either specific to each portal or lacking, represent major complicating factors.Alternate access via search-engine queries, however, first requires indexing of the data, e.g. based on technical openness and accompanied by metadata . The tecFurthermore, both quantity and quality of the available data vary. In particular, small-scale and comprehensive availability is patchy and highly heterogeneous. Despite some initiatives such as INSPIRE, finding and accessing geospatial data also remains somewhat challenging. Since health-related geographic data is relatively scarce , surveilData openness also covers aspects of participation and continued (re)usage. Although administrative data providers are encouraged to share raw data in accordance with the E-Government Act, the decision on how to use the data still remains with the authorities themselves. In addition, the absence of an explicit license prevents further use in many cases. The Geodata Access Act (GeoZG) also calls for no-cost data sharing but imposes restricted reuse that often incurs a fee. Thus, there is still a mixed landscape in terms of fees and (licensing) policies for geodata, which makes it difficult to merge data in a comprehensive manner. On the one hand, Open Citizen projects such as OSM are a valuable source in this context, not just in terms of data volume. On the other hand, this data exhibits certain disadvantages in terms of completeness, quality, topicality and validity. In contrast, official geodata is often available for administrative levels but not applicable to scientific analyses. As an example, small-scale epidemiology data is frequently provided on a postal code level, which overlaps inconsistently with administrative units. Other challenges of administrative geodata affecting spatial analyses and calling for alternative approaches are territorial reforms, data aggregations, scaling and zoning effects. In some cases, even a change of indicators may become necessary for area-wide or cross-level analyses for availability reasons.For various reasons, it is currently not possible to give an exhaustive overview of OD offerings. Given that such overviews are only able to represent a snapshot in a dynamically changing context, it would be useful to expand them on an ongoing basis. A desirable feature might be portals/overviews using software to import relevant, ideally standardized, data catalogs from decentralized resources. Metadata is a key basis and requires the use of (meta)data catalogs as well as unique identifiers for data records supplied by all data providers.The provision of OD is a joint process involving several micro and macro levels . It also implies that, besides urgently needed harmonization of legal aspects, general processes must be redesigned starting already in the context of data collection and research to initially give all data the option of openness and machine readability. In particular, the institutional and administrative bodies responsible for quality control, data management and data protection should be integrated into such strategies and processes at an early stage.schema.org metadata standard, which helps data-producing and data-providing parties structure data according to uniform vocabulary. The increased integration of Linked Data standards with the delivery process can also significantly improve data findability, accessibility and comparability in the future.Regarding the further use of research data in the course of FAIRification, discipline-specific efforts to achieve common standards, e.g. ontologies and vocabularies, would have to be pushed forward to optimize the findability and indexing of data and metadata sets in both thematic databases and general search engines. To improve the performance of the latter, Google, Yahoo, Yandex and Bing have already developed the To ensure the potential further use of data, (harmonized) legal foundations are required on the level of the federal states and municipalities. Orientation for OGD on the municipal level is currently provided by the model data catalog, which lists types and sources of OGD .Second Open Data Act and Data Utilization Act (Section 12a EGovG) [Concerning OGD, the draft law dated February 2021 on the a EGovG) could haBBSR: Bundesinstitut f\u00fcr Bau-, Stadt- und Raumforschung BImSchG: Bundesimmissionsschutzgesetz BKA: Bundeskriminalamt BKG: Bundesamt f\u00fcr Kartographie und Geod\u00e4sie BMVI: Bundesministerium f\u00fcr Verkehr und digitale Infrastruktur CC: Creative CommonsDKG: Deutsche Krankenhausgesellschaft DWD: Deutscher Wetterdienst FOIA: Freedom of Information ActFS: Federal StatesGBE: Gesundheitsberichterstattung GDI-DE: Infrastructure for Spatial Information in GermanyGeoZG: Geodatenzugangsgesetz (Geodata Access Act)GISD: German Index of Socioeconomic DeprivationINKAR: Indikatoren und Karten zur Raum- und Stadtentwicklung INSPIRE: Infrastructure for Spatial Information in EuropeOD: open dataODbL: Open Database LicenseODC: Open Data Commons\u00d6PNV: \u00d6ffentlicher Personennahverkehr (public transport)OGD: open government dataOSM: OpenStreetMapPDDL: Public Domain Dedication and LicensePKS: polizeiliche Kriminalit\u00e4tsstatistik SUF: Scientific Use FilesUBE: Umweltberichterstattung WHO: World Health Organizationhttps://code-de.org/en/CODE-DE2: https://data.deutschebahn.com/Data portal of Deutsche Bahn AG: https://opendata-esri-de.opendata.arcgis.com/ESRI Open Data: https://about.google/brand-resource-center/products-and-services/geo-guidelines/Google Maps: https://krankenhausstandorte.deHospital Locations: https://www.inkar.de/INKAR: https://www.mcloud.de/mCLOUD: https://www.mdm-portal.de/mdm Portal: https://www.statistikportal.de/de/sbeNational social welfare statistics: https://www.nvbw.de/open-dataNVBW Open Data: https://www.dwd.de/EN/ourservices/_functions/search/search_Formular.htmlOD portal of the German Weather Service: http://opendata.tursics.de/OpenDataAtlas: http://de-city.census.okfn.orgOpendata-City-Census: https://sciencemap.github.io/Open-Data-for-Health/Open Data for Health Map: https://www.opendata-oepnv.deOpen Data Platform \u00d6PNV: https://OpenSenseMap.orgOpenSenseMap: https://www.openstreetmap.orgOpenStreetMaps: https://statistikportal.thueringen.de/thonsa/SSDstart.php and http://www.gsi-berlin.info/Regional social welfare statistics (examples): http://re3data.orgRegistry of Research Data Repositories: https://sensor.community/en/Sensor Community: https://doi.org/10.5061/dryad.djh9w0w1f [Data for this article are available from the Dryad Repository: jh9w0w1f .The authors would like to thank NFDI4Health for the inspiration and knowledge sharing. This was of great value for the whole work process.The authors declare that they have no competing interests."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Scientific Reportshttps://doi.org/10.1038/s41598-023-30467-5, published online 01 March 2023Correction to: The original version of the Article contained errors in the Consortia author list of The Research Group for the Development and Evaluation of Cancer Prevention Strategies in Japan, where author name, Shiori Tanaka, was duplicated in the main author list.The original Article has been corrected."} +{"text": "In the past three decades, not only some classical MIF datasets have appeared, but also MIF technology has developed rapidly , Lytro color multi-focus image dataset and Aymaz dataset (https://github.com/sametymaz/Multi-focus-Image-Fusion-Dataset), etc. Some of these datasets were captured by professional cameras, and others were obtained by applying Gaussian blur to existing image datasets. The Multi Focus-Photography Contest dataset is an image photography competition held by the Photography Contest website. It contains 27 pairs of multi-focus images. Images in Lytro multi-focus dataset were acquired by the Lytro camera which is an all-optical camera whose imaging system employs a microlens array focused on the focal plane of the camera's main lens. The Lytro multi-focus dataset includes 20 groups of color multi-focus images and four sets of multi-source focus images. The image resolution is and the image format is jpg. The Savic dataset is collected by Nikon D5000 camera and contains 27 pairs of images. In Savic dataset, 21 pairs of images with format jpg are taken indoors, and 6 pairs of images with format bmp are used for MIF algorithm testing. In Aymaz dataset, the 150 multi-focus images are obtained by using the Gaussian blur function to locally blur some common image datasets. This dataset also contains some multiple source images of the same scene with different focal points. In addition to color multi-focus datasets, there are also some grayscale multi-focus datasets, and some images in grayscale multi-focus datasets.As mentioned above, in the past few years, a series of MIF algorithms have been developed by scholars from various countries. To test the performance of these algorithms, some classic public MIF datasets have occurred. Currently, the commonly used datasets include Multi Focus-Photography Contest dataset Normalized mutual information (NMI), which can effectively improve the stability of the MI can be found below: WP and YY performed the computer simulations. SL, WJ, and JZ analyzed the data. SL, WP, and YY wrote the original draft. WJ, YS, and JZ revised and edited the manuscript. YS polished the manuscript. All authors confirmed the submitted version.This work was supported in part by National Natural Science Foundation of China under Grant No. 62172139, Natural Science Foundation of Hebei Province under Grant No. F2022201055, Project Funded by China Postdoctoral under Grant No. 2022M713361, Science Research Project of Hebei Province under Grant No. BJ2020030, Natural Science Interdisciplinary Research Program of Hebei University under Grant No. DXK202102, Open Project Program of the National Laboratory of Pattern Recognition (NLPR) under Grant No. 202200007, Foundation of President of Hebei University under Grant No. XZJJ201909, Research Project of Hebei University Intelligent Financial Application Technology R&D Center under Grant No. XGZJ2022022, and Open Foundation of Guangdong Key Laboratory of Digital Signal and Image Processing Technology under Grant No. 2020GDDSIPL-04.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Diabetes mellitus (DM) is a metabolic disease that endangers human health, and its prevalence is exploding and younger. Stem cell-derived extracellular vesicles (SC-EVs) have a repair function similar to SCs and no risk of tumor formation, which have been widely used in the repair of DM and its complications. We aim to map the hot trends of SC-EVs for the treatment of DM and providing directions for future research.We screened all relevant publications on SC-EVs for DM from the Web of Science (Wos) during 2017\u20132021, and research trends in this field were analyzed by VOSviewer and CiteSpace.A total of 255 articles related to SC-EVs for DM were screened out according to the search strategy. China was the most productive country, followed by the USA and Italy (16 publications and 366 citations). The top five institutions with the most publications were located in Italy and China, with Turin University being the most productive. The journals Stem Cell Research and Therapy and International Journal of Molecular Sciences published most of the studies on SC-EVs for DM. ASHOK KUMAR published the majority of articles in this field, while QING LI was the most cited. Cluster analysis indicated that the current research trend is more focused on the repair mechanism and clinical translation of exosomes and their related preparations in promoting DM and its complications.In this study, a comprehensive summary and analysis of the global research trends of SC-EVs used in DM and its complications was performed. In the past 5 years, relevant high-quality publications in this field have increased significantly, and SC-EVs have a good prospect for development in the treatment of DM and its complications. Notably, DM causes 6.7 million deaths in 2021 and is responsible for at least USD 966 billion in health expenditure. Patients with diabetes in a continuous state of elevated glucose can lead to the development of various complications, such as diabetic wound, nephropathy, and retinopathy (Diabetes mellitus (DM) is a metabolic disease whose main manifestation is chronic hyperglycemia, combined with defective insulin secretion or action, and is one of the major diseases endangering human health, and its prevalence is exploding and younger . AccordiStem cells (SCs) are a class of pluripotent progenitor cells with multidirectional differentiation potential, which have been widely used in tissue regeneration engineering due to their abundant source, easy extraction and expansion, and remarkable tissue repair effects confirmed by numerous studies . SCs havvia autocrine or paracrine secretion, or act on specific and distant target cells through humoral transport, and then act directly on target cells through membrane fusion or endocytosis to participate in complex and delicate intercellular communication. Previous studies have found that SC-derived EVs (SC-EVs) have a repair function similar to SCs and no risk of tumor formation (Extracellular vesicles (EVs) are lipid bilayer vesicles secreted by most cells and are classified into exosomes (30\u2013150 nm), microsomes , and apoptotic vesicles (1\u20135 \u03bcm) based on their diameters. The secreted EVs, which contain various substances, such as lipids, proteins, and noncoding RNAs, act on adjacent target cells ormation . Notablyormation , 12.Literature is a carrier that can record scientific progress. Bibliometrics uses quantitative methods such as mathematics and statistics to study the internal connections and distribution patterns among the literature to discover the current state of research, research hotspots, and future trends in a certain field . As rese* OR SC) AND TS = (diabet* OR diabetes mellitus OR DM) AND TS = (extracellular vesicle* OR EV OR exo*) AND Language = (English) AND Publication Date = (2017-01-01 to 2021-12-31). All documents that included the above search strategy were reviewed, but letters, case reports, withdrawals, bibliography, etc. were excluded. Publicly available data sets were analyzed in this study, and ethics statement was not required. All searches were conducted on 14 January 2022 to avoid bias related to database updates.Web of Science is an authoritative academic database that has been widely used by researchers worldwide. In this study, the WoS core collection was selected as the retrieval data source of SC-EVs for DM from 2017 to 2021. The search strategy and screening process were as follows : TS = (sTwo authors (HQ and RG) independently retrieved data and excluded studies irrelevant to the collection strategy. All data, including title, keywords, authors, institutions, etc., were extracted from the WoS and were eventually included in this study. A bibliometric analysis was performed using Microsoft Excel 2021, CiteSpace V, and VOSviewer.https://www.scimagojr.com/) and eigenfactor (http://www.eigenfactor.org/index.php) websites were used to obtain the H-index and eigen factor score, respectively.Based on the data extracted from the WoS, we first analyzed the publication and citation trends of SC-EVs for diabetes and visualized them using Excel. Then, bibliometric analyses, including country and institution bibliographic coupling analysis, reference co-citation analysis, and keyword co-occurrence, were performed and visualized using CiteSpace V and VOSviewer. The newest edition of the Journal Citation Report (JCR) was used to obtain the latest impact factors (IF). The scimago journal and country rank was the most productive institution, followed by the Central South University (seven publications), Shanghai Jiaotong University (seven publications), Tianjin Medical University (seven publications), and Sun Yat Sen University (seven publications). According to citations, Shanghai Jiaotong University (306 citations) has the highest citations, followed by Central South University (242 citations) and Shangdong University (239 citations). The top 20 institutions with the most publications are listed in Then, the close and complex collaborative relationships between the different institutions were analyzed with VOSviewer. The threshold for the minimum number of documents of an organization was set at four, and the top 20 institutions met the threshold and were presented in a network map by the year of concentration of institutional publications . The resJournal distribution analysis helps to understand the hot journals in the field of SC-EVs for DM. The journal Stem Cell Research and Therapy with 15 publications, and the journal International Journal of Molecular Sciences with 15 publications, published the most studies. The journals Frontiers in Endocrinology , Cells , and Scientific Reports had seven publications each. The top 20 journals with the most publications and the impact index of the top 10 journals with the largest number of articles are presented in Then, reference co-citation analysis was performed to understand the close association between the referenced journals. The threshold for the minimum number of citations from a source was set at 20, and 210 journals met the threshold. The top 30 referenced journals were visualized in a density map, and all referenced journals were presented in a network visualization . Stem Cedoi: 10.1021/acsnano.7b07643) was a hub node in the co-citation network, followed by the article published by Li et al. in Experimental and Molecular Medicine in 2018 (doi: 10.1038/s12276-018-0058-5). To better understand the scientific frontiers of the field on SC-EVs for DM, we analyzed the references using the burst detection function (the minimum duration threshold was set as one) in CiteSpace. The top 25 references with the strongest citation bursts are presented in The most productive authors with the highest number of publications and citations from 2017 to 2021 are listed in A title cluster analysis was implemented to generalize the references in the co-citation network to understand the frontier directions. The references in the co-citation network were divided into nine different clusters, including diabetic wound, human umbilical cord, kidney diseases, hypothesis evidence, cell\u2013cell communication, cell-derived EV, engineered exosome, key miRNA, and tissue repair . Based oKeywords represent the central topic of a publication, and keyword co-occurrence analysis helps to systematically understand the hot topics and current progress of SC-EVs for diabetes and their intrinsic connections. VOSviewer was used to analyze keywords, and the threshold was set at a minimum of five occurrence of a keyword in the titles and abstracts of the included publications. A total of 112 keywords were identified and were mainly divided into seven different clusters: EVs, exosomes, angiogenesis, SCs, mesenchymal SCs, diabetes, and oxidative stress . In the Diabetes mellitus is a metabolic disease caused by islet dysfunction or insulin action disorder, and its incidence has increased steeply in the last decade . Long-tePublished research in the field of diabetes continues to increase year by year, and the intervention and management of diabetes have attracted widespread attention . In receBibliometrics is now widely used in various fields of global research, helping researchers to gain an intuitive and systematic understanding of a field and to identify significant scientific achievements and future research hotspots. Statistics on the number and citation frequency of SC-EVs used in DM publications show that the number of publications and the frequency of citations have increased significantly over the past 5 years, indicating that this field is a current research hotspot . NotablyTo better understand the national and regional contributions in this field, a national distribution analysis of publications was performed. China ranks first in the world in terms of total number of publications and citations, but the average citation rate in China (22.61) is lower than that of the USA (23.34), Iran (29.9), and Italy (23.34), which means that China still needs to improve the quality of publications . A recendoi: 10.1038/s41581-018-0023-5) had the largest number of citations in the co-citation network, and Ying Wang was the corresponding author (doi: 10.1021/acsnano.7b07643) and Li (doi: 10.1038/s12276-018-0058-5) are hub nodes in the co-citation network .The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "To the Editor,Over the years, the disease burden of communicable and non-communicable diseases among developing countries has been on the rise. Traditionally associated with developed countries, developing countries have seen an unprecedented increase in the past years in non-communicable diseases such as diabetes, cardiovascular diseases, obesity. It was predicted that by 2020, NCD's will be responsible for about 70% of the global burden of disease, causing 7 out of 10 deaths in developing countries [A review of literature shows that developing countries like Bangladesh, India, Nepal, Pakistan and Sri Lanka spend less that 4% of their GDP on healthcare . This reThe rising cost of healthcare is a grave problem for resource depleted countries in addition to the rising burden of diseases. It not only affects the quality of care being provided but also has led to the rationing and limiting of healthcare services . FinanciThis situation calls for an immediate remedial and re-evaluation of our policies regarding healthcare. For developing countries lack of healthcare expenditure, allocation of budget and inadequate resource utilisation is a major problem . DevelopA greater stress on health maintenance and disease prevention is the ideal and efficient approach to counter healthcare budget and cost constraints. There also needs to be a shift of focus towards primary health care, which is a more cost-effective way of delivering health services to a wider population . GloballFunding of research programs and labs to come up with new diagnostic tests and medications in an attempt to minimize costs can also be done. Training of paramedics, nurses, and pharmacists to play a more active role in healthcare delivery can be fruitful in improving outcomes and prevent negligence ,15.Overall, the priority of the developing countries need to be focused around adopting a more humanised approach towards healthcare. Healthcare should be equally accessible to the poor and the rich with no discrimination in the quality of services. A bitter reality is that while the developed world progresses, developing countries health indicators worsen perpetuating the cycle of poverty and illness .Not commissioned, externally peer reviewed.None.Not applicable.Not applicable.Faizan Fazal: Study conception, write-up, critical review and approval of the final version.Tayyaba saleem: Study conception, write-up, critical review and approval of the final version.Mohammad Ebad Ur Rehman: Study conception, write-up, critical review and approval of the final version.Tehseen Haider: Study conception, critical review and approval of the final version.Abdul rauf khalid: Study conception, write-up, critical review and approval of the final version.Usama Tanveer: Study conception, write-up, critical review and approval of the final version.Haris Mustafa: Study conception, write-up, critical review and approval of the final version.Junaid Tanveer: Study conception, critical review and approval of the final version.Arooba Noor: Study conception, write-up, critical review and approval of the final version.1.Name of the registry: Not applicable2.Unique Identifying number or registration ID: Not applicable3.Hyperlink to your specific registration (must be publicly accessible and will be checked): Not applicableFaizan Fazal.Mohammad Ebad Ur Rehman.The authors have no conflicts of interest."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "In recent years, with the vigorous development and application of Artificial Intelligence (AI), the application of AI in education is becoming more and more extensive. This study makes a theoretical analysis of AI-Intellectualized Information Technology (IT). Discrete Cosine Transform (DCT)-Based Speech Recognition (SR) and Genetic Algorithm (GA)-Based Image Recognition (IR) are used to analyze the College Ideological and Political Education (IAPE). The research findings prove that the advantages of integrating AI-intellectualized IT on College IAPE outweigh the disadvantages. The improvement of technological development, which accounts for 71.17% of undergraduate gains, is the most significant, and the smallest gain is technology coverage, which is 36.80%. Overall, 57.21% are interested in new technology, and the students' enthusiasm accounts for 30.77%. Most of the students focus on the innovation performance of technology, accounting for 75.92%. With an average influence of 89.04% on undergraduates, technology has the largest impact, followed by 85.78% on students with masters or higher degrees. The largest impact of diversified teaching methods for all students is 62.48%. This study provides some reference values for AI-intellectualized IT research and analysis, as well as students' IAPE. Recently, with the continuous development of Artificial Intelligence (AI) and Information Technology (IT), major colleges and universities have also developed AI and IT-related majors one after another, making AI one of the hot topics . Many AIThe twenty-first century has witnessed accelerated integration of various disciplines, emerging new domains, and extension of scientific frontiers . ConsequLi et al. underlinThis study provides a theoretical analysis of AI and IT in the development of the ideological and physical education of college students. Using the Discrete Cosine Transform (DCT)-Based Speech Recognition (SR) and Genetic Algorithm (GA)-Based Image Recognition (IR) in students' IAPE, the advantages and disadvantages of AI-intellectualized IT on College IAPE are examined and gains analysis and its impact on education are provided. The present study has certain reference significance for the research and analysis of AI-intellectualized IT and students' IAPE.The rest of the manuscript is organized as follows. AI is a new technical science that studies and develops theories, methods, technologies, and application systems for simulating, extending, and expanding human intelligence . It aimsAs shown in As a general technology, AI can be widely used in various fields, including its integration with education. AI is the core of many virtuous interaction circles, including business data analysis, NLP, SR, machine reasoning, computer vision, robots, and sensors. Surely, applying these technologies in education will help the reform of higher education. C(n) is the Mel-scale cepstrum parameter of order L. L refers to the order of Mel Frequency Cepstral Coefficient (MFCC). Usually, order 12 can represent the acoustic characteristics. s(m) denotes the signal after framing, and N stands for the size of the frame. M means the number of triangular filters, and the transfer function of each band-pass filter is calculated using the following equation (Hm(k), (0 \u2264 m \u2264 M) represents several band-pass filters set within the sound spectrum range. f(m)is the center frequency of each filter, and in (f(m) can be calculated:fl and fh represent the lowest and highest frequency in the filter frequency range, respectively. N is the length of FFT. f shows the sampling frequency. The corresponding relationship between frequency and Mel-scale is linear below 1,000\u2009Hz and logarithmic above 1,000\u2009Hz. Equations caquations and 5) (1)Cn=\u2211mf is the frequency, and the unit is Hz. Equation (s(m) is the logarithmic energy output by each filter bank and X(k) indicates the time domain signal. Logarithmic processing is performed in the frequency domain, and the logarithm is inversely transformed to obtain the MFCC coefficient, as expressed in (In and 5),,5), f isEquation reflectsessed in and 8)..f is theEquation is a logThen, the extreme of and 8) 8) is cad(t) represents the t-the first-order difference and c(t) indicates the t-th Cepstrum coefficient. Secondly, using the automatic analysis and application of AI-intellectualized IT, according to IR and SR results, this study analyzes the teaching process of College Students' IAPE and the changes in students' ideology and political views. The video is segmented according to the video image changes in the IR field. Whether each segment belongs to the IAPE teaching process or video playback is judged simultaneously. Then, the video image segmentation data are recorded and stored in the database. IR can be performed through GA [f represents the fitness function. The sample size is N. f(xi) is the fitness of the sample xi. The SR process converts image files into audio files, extracts sound features, and stores them in the feature list. Then, feature matching is carried out to complete cluster analysis. Finally, the process of IAPE is completed, and the teaching process is analyzed according to the principles of College IAPE. In equation , d(t) rerough GA . First, According to the relevant characteristics and technical requirements, in College IAPE, AI-intellectualized IT can not only achieve the knowledge expression of traditional IAPE but also use the Expert System Shell (ESS)-based simple expert system to let students experience the application of AI-intellectualized IT in learning IAPE. Then, it analyzes and solves students' ideological and political problems. In particular, an Intelligent Computer-Aided Instruction (ICAI) can be used to apply AI-intellectualized IT to the college IAPE system based on a computer medium and cognitive science. As plotted in The expert system structure in College IAPE teachers should well manage AI-intellectualized IT . AI techAI-intellectualized IT can change students' learning mode but can never alter the ultimate goal of learning. In other words, learners should be well aware that AI, as a technological approach, can assist the learning process but can never replace human intelligence. After all, the goal of learning is to improve the student's ability. The advantages and disadvantages of applying AI-intellectualized IT to UGRDs' IAPE have been evaluated from the following six performance indexes including data comprehensiveness, product performance, technology openness, students' adaptability, enthusiasm, and technology dependence. The concerns and gains of UGRDs' IAPE integrated with AI-intellectualized IT can be evaluated from comprehensive ability, innovation, development, and coverage. As indicated in According to the educational background, college students are divided into UGRD, postgraduate, and higher grades. The impact of IAPE on UGRDs in integrating AI-intellectualized IT is assessed from the diversification of teaching methods, accuracy of teaching plans, personalized lesson content, intelligent data resources, scientific evaluation methods, dynamic teaching process, intelligent teaching environment, and reducing classroom burden. In Students must participate in the creation of ethnic education and embrace appropriate norms, policies, values, and processes for academics to continue to progress positively for intellectual Ideological and Political Learning exploration and study. This study explores the integration of AI-intellectualized IT into College Students' IAPE, the advantages and disadvantages of College IAPE integrated with AI-intellectualized IT along with concerns and gains analysis, and the impact on education through AI-intellectualized IT. The results verify that the advantages of integrating AI-intellectualized IT with IAPE in colleges and universities outweigh the disadvantages. The biggest gain for UGRDs is the improvement of technology development, 71.17%, followed by students' comprehensive ability improvement, which is 65.53%, and the smallest yield is the coverage of technology, which is 36.80. Most students focus on the innovation performance of technology, accounting for 75.92%. Technology has the greatest impact on UGRDs, with an average impact of 89.04% and 85.78% with postgraduates or above. Overall, the largest impact of diversified teaching methods is 62.48% on all students. This study has specific reference significance for the research and analysis of AI-intellectualized IT and College Students' IAPE. However, because of the limitations of AI-intellectualized IT in College Students' IAPE, initiatives are required to understand the limitations of AI and learn to use technology correctly in a reasonable scene."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Mycotoxin risk in the feed supply chain poses a concern to animal and human health, economy, and international trade of agri-food commodities. Mycotoxin contamination in feed and food is unavoidable and unpredictable. Therefore, monitoring and control are the critical points. Effective and rapid methods for mycotoxin detection, at the levels set by the regulations, are needed for an efficient mycotoxin management. This review provides an overview of the use of the electronic nose (e-nose) as an effective tool for rapid mycotoxin detection and management of the mycotoxin risk at feed business level. E-nose has a high discrimination accuracy between non-contaminated and single-mycotoxin-contaminated grain. However, the predictive accuracy of e-nose is still limited and unsuitable for in-field application, where mycotoxin co-contamination occurs. Further research needs to be focused on the sensor materials, data analysis, pattern recognition systems, and a better understanding of the needs of the feed industry for a safety and quality management of the feed supply chain. A universal e-nose for mycotoxin detection is not realistic; a unique e-nose must be designed for each specific application. Robust and suitable e-nose method and advancements in signal processing algorithms must be validated for specific needs. Mycotoxins are one of the largest safety risks for the feed/food chain, with a negative impact on animal and human health, economy, and international trade of feed and food commodities ,3,4,5,6.After a brief survey on mycotoxin contamination in animal feed, this review provides an overview of the use of e-nose as an effective tool for rapid mycotoxin detection and management of the mycotoxin risk at feed business level.Fusarium, Aspergillus, Penicillium, and Claviceps spp. mycotoxins produced by represent the main contaminants of the feed supply chain, with important impact on animal health, productivity, and feed/food safety . O. O132]. E-nose represents a powerful tool in the feed chain for quality and safety control and monitoring. E-nose offers potential as a rapid and cost-effective diagnostic tool for mycotoxin contamination screening at the market entry level. However, before e-nose laboratory-based assays can move from research into the feed industry and become a reality, we must face and overcome several challenges to improve e-nose performance.The future challenges are: the sensor materials, data analysis, pattern recognition systems, and a better understanding of the industrial needs related to safety and quality control of the feed supply chain. A universal e-nose for mycotoxin detection is not realistic; a unique e-nose must be designed for each specific application. Limitations still exist regarding sensitivity and selectivity of sensors. The major drawback is represented by sensors\u2019 sensitivity to environmental conditions, particularly humidity and temperature. Improved modelling, correlation between chemical markers and sensor responses, and robust and suitable e-nose methods and advancements in signal processing algorithms must be validated for specific needs. In the field of mycotoxin co-contamination detection, the predictive accuracy of the e-nose models is still limited for industrial applications in a real context. Last, but not least, specific sampling models must be carefully selected to enhance the accuracy of e-nose analysis."} +{"text": "Bladder cancer is one of the most common urological cancers. Non-muscle invasive bladder cancer (NMIBC) accounts for about 75-85% of all newly diagnosed bladder cancers. Globally, there are many NMIBC-related publications. However, a bibliometric analysis of these publications has not been performed.This study aims to systematically analyze and visualize NMIBC-related publications through bibliometrics, and to reveal identified topics, hotspots, and knowledge gaps in related fields.Based on the Web of Science core collection database, we firstly analyzed the quantity and quality of publications in the field of NMIBC, secondly profiled the publishing groups in terms of country, institution, author\u2019s publication and cooperation network, and finally sorted out and summarized the hot topics of research.This bibliometric analysis was conducted from 2001 to 2022. The analysis identified 2,185 articles and reviews, which were published in 402 journals. The number of publications and citations on NMIBC-related research has steadily increased over the last two decades. Furthermore, academic institutions in Europe and the United States play a leading role in NMIBC research. The country, institution, journal, and author with the most publications were the United States (559), Radboud University Nijmegen (88), Urologic oncology: Seminars and Original Investigations (141), and Witjes J (74), respectively. The most frequently used keywords were Bladder cancer (793), Recurrence (671), Urothelial carcinoma (593), Progression (523), Bacillus-calmette-guerin (411), Transitional-cell carcinoma (401), Carcinoma (366), Risk (297), Transurethral resection (286), and Non-muscle-invasive bladder cancer (280).More and more scholars are devoted to the research of related NMIBC. This bibliometric analysis revealed that the main research topics and hotspots in NMIBC included pathological staging, clinical diagnosis and treatment, and bladder perfusion. Bladder cancer is the most common malignancy of the urinary system. Its incidence is the first among malignancy of urinary system, and it is also the tenth most common cancer in the world . ApproxiThe pathogenesis of NMIBC is still unclear. However, bladder cancer is associated with various risk factors, including smoking, gender, occupation, and age . CurrentBibliometrics is the quantitative analysis of literature, which is widely used for evaluating research trends and hotspots in various fields . VOSviewHundreds of original articles and reviews on NMIBC are published each year. However, so far, there is no systematic analysis of NMIBC-related publications. Therefore, this study aimed to systematically analyze and visualize NMIBC publications over the past 20 years using related bibliometric softwares, such as VOSviewer and CiteSpace. Furthermore, the study aims to summarize the achievements attained in this field, understand the research direction and hotspot areas, and provide a reference for future studies.https://www.webofscience.com/wos/woscc/basic-search) and hence, it is deemed to be exempted from the Institutional Review Board approval.The present study did not involve any human subject participation, and it was entirely performed using the bibliometric data retrieved from the Web of Science database AND \u201cbladder cancer\u201d (Topic) and Article or Review Article (Document Types) and Book Chapters (Exclude-Document Types).The data were downloaded and analyzed by two researchers respectively to assure the accuracy of data and the repeatability of the research. Microsoft Excel 2019 and GraphPad Prism 7 were applied to analyze the targeted files and exported the line charts and tables of top-cited or productive countries/regions, institutions, authors, journals, references, keywords.2) was used to predict the relationship between publication year and publication output to compare the degree of agreement between the predicted results and the actual occurrence. The closer R2 is to 1, the better the fitting degree of the regression line to the observed value is; otherwise, the worse it is. At the same time, the 2021 version of Impact factor (IF) and Journal impact factor quartile (JIF) quartile, as important indicators to measure the scientific value of research, were also included in the analysis.The test of fit is a visual analysis software widely used in scientific papers. It is based on scientometrics data and information visualization technology. Further, it presents the knowledge structure by analyzing the underlying knowledge, patterns, and distribution of the literature is a bibliometric analysis software for mapping knowledge. It can be used for co-word analysis, co-citation analysis, coupling analysis of documents, and to visualize the results in NMIBC-related research, reflecting the increasing interest in this research field had the highest number of publications , followed by China , Italy , Germany , and the Netherlands Figure\u00a03VOSviewer was used to analyze the cooperation of different countries. The line between nodes indicates the co-authorship between countries; the thicker the line, the stronger the cooperative relationship. The results showed that the USA, China, and Italy had more cooperation with other countries. However, cooperation between the other countries was weaker Figure\u00a04A total of 2,978 institutions were involved in publishing NMIBC-related papers. The top five institutions with the highest number of publications were Radboud University Nijmegen (88), Medical University of Vienna (85), University of Texas MD Anderson Cancer Center (77), Autonomous University of Barcelona (63), and Charles University Prague (48) Table\u00a02.The author co-occurrence analysis identified the core authors in NMIBC-related research and the strength of collaboration between authors. Co-cited analysis means that when two authors or papers are cited by a third author or paper at the same time, the two authors or papers have a co-cited relationship. This analysis revealed a total of 9,979 authors and 23,062 co-cited authors. Among them, Witjes J (74), Shariat S (70), Kamat A (58), Roupret M (40), and Burger M (37) had the highest publications , BJU International (119), European Urology (100), World Journal of Urology (93), and International Journal of Urology (51). The most cited journals were European Urology , BJU International , Urologic oncology: Seminars and Original Investigations , World Journal of Urology , and Nature Reviews Urology Table\u00a04.Analysis of the co-cited journals showed that 4,863 journals were co-cited. The top five co-cited journals were Urology Journal , European Urology , BJU International , UROLOGY , and Journal of Clinical Oncology Table\u00a05.A total of 2,185 references were obtained, of which seven references Babjuk et\u00a0al., , Kamat eThrough keyword co-occurrence and burst analysis, we can identify the changing trend of research topics over time. A total of 5,422 keywords were obtained. The top ten keywords obtained were Bladder cancer (793), Recurrence (671), Urothelial carcinoma (593), Progression (523), Bacillus-calmette-guerin (411), Transitional-cell carcinoma (401), Carcinoma (366), Risk (297), Transurethral resection (286), and Non-muscle-invasive bladder cancer (280), in situ (16.37), In addition, a total of 17 keyword-burst analysis results were obtained. From 2005 to 2015, the research hotspots in the NMIBC field were mainly randomized clinical trials and meta-analyses, which focused on cancer staging, and white light cystoscopy. However, from 2016 to 2022, the research hotspots in the NMIBC field are mainly focused on efficacy and safety. The three salient words with the highest strength were transitional cell carcinoma (30.5), stage Ta (18.52), and With the advent of big data, researchers need to understand the developments within NMIBC-related research. The bibliometric analysis uses visualization softwares, such as VOSviewer and CiteSpace, to comprehensively analyze the existing literature, understand the research trends, and predict future research hotspots . This bi2 = 0.8368).The highest publications were made in the past four years, with more than 200 publications being made each year. Most of the studies were funded by the National Science Foundation and multinational pharmaceutical companies, indicating that research in this field contributes to the national scientific and technological knowledge (Tis), or with limited invasion of the lamina propria (T1) TNM staging system. These guidelines define NMIBC as noninvasive papillary UCB (Ta), carcinoma ria (T1) . Furtherria (T1) . At presria (T1) .The keyword analysis identified white light cystoscopy (WLC) and photodynamics as the main terms for examination and treatment modalities. The photodynamic-related keywords included Photodynamic Diagnosis (PDD), Photodynamic Therapy (PDT), and photosensitizer 5-aminolevulinic acid (5-ALA). The WLC-assisted endoscopy is currently the standard method for endoscopic detection and resection of bladder cancer . This meMoreover, the keyword analysis identified intravesical instillation, especially with BCG, as one of the main categories. The treatment of NMIBC often combines TURBT with intravesical instillation to reduce the recurrence rate. Intravesical instillation is often carried out using chemotherapy drugs and immunotherapy agents. BCG is the main immunotherapy agent employed in intravesical instillation of NMIBC patients after surgery. In March 2020, the European Association of Urology (EAU) recommended that BCG treatment after TURBT was more effective than TURBT alone or TURBT plus intravesical chemotherapy in preventing tumor recurrence and halting disease progression . In the Moreover, the keyword analysis revealed that research on NMIBC was more concentrated in the European region and institutions, probably due to the high incidence of NMIBC in developed countries, such as Europe and North America . FurtherThe significance of our research is as follows: 1. The countries/regions, institutions, authors, journals, references, keywords and other elements of NMIBC related literature in the past 20 years were clearly displayed. 2. Through the collation of data sets, readers and experts in relevant fields were helped to understand the development process, research status and knowledge hotspots of NMIBC. 3. Future research directions of NMIBC were further provided for reference. But, there are still some limitations to our study: 1. Recently published articles may have low citations due to the limited time available for citations, and thus the study may be prone to research bias. 2. This study only included articles and reviews published in English, which may have overlooked some of the literature. 3. With the rapid development of big data, this study may have a short timeframe and needs to be updated regularly.The number of published papers in the NMIBC field has been growing steadily since 2005, with over 200 papers published annually in the past three years. The USA and Europe are leading in research on NMIBC. However, there is a need to strengthen collaboration relationships, especially in developing countries. The top five cited and co-cited journals were mostly in Q1 and Q2, reflecting that research on NMIBC was of high quality. Furthermore, this bibliometric analysis revealed that the main research topics and hotspots in NMIBC included pathological staging, clinical diagnosis and treatment, and bladder perfusion.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.SD and JW designed the study. SD, ZY, and LX conducted the literature search. SD, FM, and LW analyzed the data and wrote the paper. JW and ZX approved the final manuscript. All authors contributed to the article and approved the submitted version.China Postdoctoral Innovative Talent Support Program (BX20220047); Young Talent Support Project of Beijing Association of Science and Technology (BYESS2022182); Young Talent Support Project of Chinese Association of Chinese Medicine (CACM-2021-QNRC2-B04).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "In times of global crisis like the present Covid-19 pandemic, digital technology is rapidly conquering the health and mental health & care sector, speeding up e-Mental Health (eMH) implementation on a regional, national and global scale. Making this an organized move, guidance and regulation, legislation and training, but basically also awareness and acceptance building need to ensure the use of efficient, safe and high-quality eMH products and services. Special attention needs to focus on broadening public and professional eMH literacy, providing needs-tailored approaches for target groups, and training mental health workforce and services. Guidance, evaluation and involvement of relevant stakeholders should help to identify how citizens will best benefit from eMH&Care in its various forms. The Transnational Policy for e-Mental Health, a guidance document for European policymakers and stakeholders has been developed by the Interreg-funded eMEN project (www.nweurope.eu/emen) in six EU countries to promote implementation of high-quality eMH & care across NW-Europe. Project partners from Belgium, France, Germany, Ireland, the Netherlands and the UK contributed to product and policy-guidance development, promoting communication and research. eMEN is currently continuing its work in the Interreg-funded Capitalisation phase to scaling up the implementation of eMH&Care. The Transnational Policy within the scope of national information and training sessions on eMH will be promoted for action planning and implementation by policymakers and stakeholders at the national level. Further meetings will also take place at the European level to promote and support implementation of eMH&Care in NW-Europe and beyond.No significant relationships."} +{"text": "Strong evidence demonstrates the long-term influence of stress and well-being on psychological, social, and physical health outcomes across the lifespan. Because of this, stress and well-being measures have been added to nearly all of the International Family of Health and Retirement Studies. However, this newly available data has not been compared cross-nationally or within-country to unpack how culture influences these important predictors of healthy aging. Using the Gateway to Global Aging Data, which provides harmonized data from the Health and Retirement Study and its sibling nationally representative studies, levels of self-reported stress and well-being are compared across 30 countries. Data come from the following studies: HRS, ELSA, SHARE, TILDA, CHARLS, KLoSA, MHAS, and JSTAR. We used data from the latest study wave for which the relevant survey was implemented. Average age of participants across studies is 67 and 55% are women. Initial analyses show stressor specific findings such as participants in Korea reported greater work stress than participants in Japan, England, the United States, and across Europe, and the United States reported higher loneliness than China and England, but not higher than Ireland. Reporting cross-national and within-country variation in these measures will be generative in pointing to new research directions for understanding how culture influences health and aging trajectories."} +{"text": "CAPABLE is an evidence-based 4-to-6-month program that improves daily function and the home environment based on the older adult\u2019s goals. Study aims were to: (1) understand context and readiness factors in implementation, (2) identify barriers and facilitators, and (3) examine the utility of two frameworks. The unit of analysis was the organization. We employed the CFIR and RE-AIM frameworks for this qualitative study, using multiple data sources and traced the implementation of 40 organizations over 3 years. Leadership support, perceived value of the program, initial funding, strategic importance, and program champion were common supportive factors. Common challenges included difficulty with recruitment, staffing and infrastructure readiness and sustainability of funding. This study indicates readiness, technical support, and resources needed for implementation and sustainability of CAPABLE and suggests external environmental supports needed. It offers a practical way to monitor, evaluate, and report on ongoing implementation of evidence-based programs."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Funding: This project was funded by The Deputyship for Research & Innovation, Ministry of Education in Saudi Arabia, the project no. (IFKSURG-2-940).Acknowledgments: The authors extend their appreciation to the Deputyship for Research & Innovation, Ministry of Education in Saudi Arabia for funding this research work through the project no. (IFKSURG-2-940).In the original publication , the funThe authors state that the scientific conclusions are unaffected. This correction was approved by the Academic Editor. The original publication has also been updated."} +{"text": "The Special Issue \u201cDynamics and Processes at Laser-irradiated Surfaces\u201d is dedicated to the 70th birthday of J\u00fcrgen Reif, retired full professor, former Chair of Experimental Physics II of the Faculty of Physics of the Brandenburg University of Technology Cottbus\u2014Senftenberg in Germany.This Special Issue was organized by one of his former PhD students and collaborators (F.C.), one of his collaborators (S.V.), and one of his external colleagues (J.B.) in recognition of J\u00fcrgen Reif\u2019s long-lasting scientific contributions and research lines in the fields of photonics, semiconductor technology, optical spectroscopy, surface dynamics, in situ measurement techniques, experimental and theoretical investigations of laser\u2013matter interaction, applications of surface functionalization through laser-induced micro- and nanostructures, laser processing of polymers, numerical modeling of surface processes, etc.Molecules and Nanomaterials, published in a separate branch for each journal [Because these scientific areas lie at the boundaries between nonlinear optics and spectroscopy, surface science, solid-state physics, materials science and nanotechnology, and in honor and recognition of Prof. Reif\u2019s scientific contributions, this Special Issue was simultaneously organized in the two journals journal ,2.For more than 25 years, apart from educating and qualifying university students for the future, the work of Prof. Reif has been very actively devoted to the study of fundamental mechanisms, dynamics, and applications of nano- and micrometer-scale laser-induced periodic surface structures (LIPSS), with a special focus on self-organization processes . His conOur Special Issue attracted high-quality contributions from both academics and from industry and finally bundles together 1 review paper of J\u00fcrgen Reif , 1 perspNanomaterials and Molecules for their professional support and guidance.Finally, the Guest Editors would like to express their sincere gratitude to all authors and reviewers of this Special Issue for their contribution to this Special Issue and to the editorial staff of"} +{"text": "Nature Microbiology 10.1038/s41564-022-01268-9, published online 10 November 2022.Correction to: In the version of this article initially published, Bradley Pickering, Oliver Lung and Finlay Maguire were not denoted as equally contributing to the work, and Samira Mubareka and Jeff Bowman were not denoted as jointly supervising the work. The changes have been made in the HTML and PDF versions of the article."} +{"text": "In the current era of digital economy, the role of information communication and technology (ICT) and economic complexity are important for controlling environmental unsustainability and formulating policies to deal with ecological concerns. However, the relationship between digital economy and environment has been studied widely; nevertheless, the relationship between ICT-based digital economy, economic complexity, and ecological footprint has not been studied extensively. Therefore, the aim of current study is to fill the existing gap by investigating the relationship between ICT, economic complexity, and ecological footprint in the case of G-seven economies. Furthermore, the past research studies were usually based on carbon emissions to measure environmental sustainability, while this study fills the gap using ecological footprint as a proxy for environmental degradation. By using the panel data over the period of 2001\u20132018 for G-seven economies, this study performs first-generation as well as second-generation unit root testing methods. Findings of both Pesaran\u2019s and B&P\u2019s cross-sectional dependence testing approaches confirm the presence of cross-sectional dependence across all G-seven economies. The empirical findings of cointegration (Pedroni and Kao) tests verify a stable long-run association between ecological footprint, ICT import, ICT export, economic complexity, economic growth, and other control grouped variables. The empirical evidence obtained from the fully modified OLS model suggests that ICT export, economic complexity, and economic growth enhance the intensity of ecological footprint, while ICT import, research and development (RD), and trade are helpful in reducing ecological footprint in G-seven economies. These empirical findings obtained are verified by pooled mean group-ARDL (PMG-ARDL) methodologies and confirm that there is no inconsistency in the results. On the basis of these results, some policy implications for ecological footprint, ICT, and economic complexity are discussed. The challenge of climate changes and environmental degradation are not national issues now, but these are a global challenge and cross the national border . Issues Although the achievement of the highest economic growth is the end goal of any country, developed countries are mostly concerned about their environmental sustainability . In contThe developed economies, such as the G-seven group, led a massive increase in economic growth during the recent decade. As a matter of fact, G-seven economies hold about 60% of global wealth . In contrast, developing economies are becoming emerging economies by improving their human capital, energy supply structure, infrastructure, and economic growth. Among them, the economic production and financial output of China and India were about USD 30,000 billion, which were far more than as compared to the United States and all European Union (EU) countries in 2017\u20132018 [In the current time of digitalization, the contributing role of ICT on environmental quality and economic growth has been discussed by numerous environmental economists. In the path of economic growth, ICT contributes to economic growth in real terms in the following three ways. First, the emerging of ICT cuts the total factor costs (TFC) by providing good direction of communication with lower transaction costs, for instance, online business, online banking, online buying and selling, E-commerce, and numerous applications of smartphones . Second,However, the contributing role of ICT is not limited to real economic growth; the enormous infrastructure development related to ICT and other macro and micro uprising can also control the environmental performance by means of the cost\u2019s upshots, consumption, and their substations. Primarily, technological infrastructure based on ICT improves utilization of energy consumption, while collecting, installation of machineries, and distribution of these machines and equipment generates toxins, which degrade the ecological system ,15,16,17In current prevailing economic structure, economic complexity index (ECI) has been considered with widespread attention among many social and environmental scientists, because it is also a most consistent and valid indicator of economic growth, as with ICT . On the In the context of the above discussed arguments, the current study investigates the impact of information communication and technologies (ICT) and economic complexity (ECI) on ecological footprint (EcoFP). To do so, we used fully modified ordinary least square (FM-OLS) and pooled mean group-autoregressive distributive lag (PMG-ARDL) regressions models over the time spanning from 2001 to 2018 for the G-seven group of countries . This investigated group of countries was originally founded in the early 1970s, while, now, they are economically well-developed and technologically advanced nations. In early 2018, these economies collectively comprised approximately 60% of the global wealth for a total of USD 317,000 billion, while their share of nominal GDP was about 45.8%, purchasing power parity-based GDP approximately 31%, and they comprised 770 million people or 10% of the world\u2019s population . So, it The G-seven group countries are investigated for a few reasons. First, these economies hold 29.9 percent of global GDP and based on digital economy but, at the same time, they have a huge share of global energy consumption; consequently, G-seven are responsible for environmental unsustainability . Second,On the basis of the above-mentioned discussion, the current study examines how digital economy, in terms of ICT, affects environmental sustainability in terms of EcoFP. Then, in the context of economic complexity (ECI), it makes a great effort to investigate the direct and indirect controlling mechanisms of digital economy on EcoFP. Furthermore, the possible effect of ECI is explored, along with the influencing mechanism of ICT on EcoFp relevant to ECI also beingverified. The results obtained from the current study can serve as a theoretical guide for testing how digital economy in terms of ICT and economic complexity influence the growth of EcoFP. Temporarily, the results also provide empirical support regarding digitalization of economy, a skills- and knowledge-based industrial process, and enhancement of environmental sustainability in many developed and developing economies.In this context, we seek to report the following question: what impacts do ICT-based digital economy and economic complexity have on ecological footprint in the case of G-seven economies? We seek to empirically answer the above-mentioned research question by aiming to examine the relationship between ICT, economic complexity, and ecological footprint in G-seven economies by incorporating many control variables. The current research adds to the existing literature in five ways: first, as far as we know, there are hardly any studies that have examined the relationship between ICT-based digital economy, ECI, and EcoFP for G-seven economies. Second, as compared to other papers in the existing literature, in this study, we inspect how ICT (two types of ICTs: ICT-exported goods and ICT-imported goods) and economic complexity influence EcoFP . Third, besides the wide-ranging proxies of environmental degradation (EcoFP) and digitalization of economy (ICTs and ECI), we have incorporated economic growth (GDP), research and development (R&D), foreign direct investment (FDI), trade ratio, and population as important control variables to avoid biasness. since the early work of Grossman and Krueger , where tIn the recent existing literature of environmental economics, some empirical studies have utilized EcoFP as a measure of environmental sustainability. EcoFP supports us to recognize well both direct and indirect effects of consumptions and production on the environment ,5,35. AfHowever, in this era of digitalization, the significance of ICT and ECI on development has been broadly deliberated in the last few years. According to Schumpeter and the Hypothetically, one school of thought has appeared as a basis to assume the linkage between digital economy (adaptation of ICT) and environmental sustainability. One is that the modernizations of an ecological system which supports that digital economy in terms of ICT can enhance economic, structural transformation, improved technologies, changes in the industrial process, and, therefore, environmental regulations and mitigation. The emergence of ICT will eventually improve environmental quality and sustainability in terms of low levels of carbon emissions and ecological footprint ,46. AnotThe emergence and advancement of ICT progress have different effects on climate change and our ecosystem. It is associated with the formation of environmentally friendly goods and services and consumption of cleaner energy. Furthermore, technological advancement is opening up new apparatus with low nonrenewable energy consumptions and power supply units that are based on renewable energy consumption and, therefore, mitigate environmental pollution. The association between ICT and EcoFP can be studied from different viewpoints: real income , human cApart from ICT, measure of economic complexity, ECI, is positively related with environmental degradation . AccordiThe role of digital economy (ICTs and economic complexity) is not only restricted GDP growth (economic); the enormous development of ICT-related economic structure and related knowledge- and skill-based industrial structure can also influence the environment by means of their consumption pattern, cost, and substitution effect. At the initial stage, ICT-based technology, and ECI-based machines and equipment increase the demand for energy consumption . ParticuHypothesis\u00a01.Information communication and technology impacts on ecological footprint.Hypothesis\u00a02.Economic complexity has an impact on ecological footprint.Furthermore, from the above-mentioned discussion and related literatures, we observed that ICT and ECI both have the ability that can increase the quality of environment or increase environmental unsustainability, depending on their level of diffusion in an economy, time period of analysis, and data and method that are incorporated. To the best of our knowledge, only a small amount of empirical research scrutinized the impact of ICT and ECI on EcoFP in the context of developed countries, especially G-seven economies. To the possible extent, the existing literature has overlooked some important environment-influencing factors, such as research and development (RD), that might be some foundation of misspecification and biasness. In addition, to our knowledge, the current research is the first study of its kind that examines the linkages between ICT, ECI, and environmental degradation in G-seven economies over the period of 2001\u20132018. Our study aims to fill the existing research gap in the literature. The empirical findings obtained from the analysis of the study would deliver initial caveats for policymakers of the targeted region.exp and ICTimp) as proxies for digital economy; ICPexp measures the percentage of total ICT-related exported goods, while ICTimp measures the percentage of total ICT-related imported goods, ECI presents a country\u2019s composition expression of the production process by inclusion of the knowledge on their diversity (goods exports) and economic growth measure as per capita GDP (USD). Likewise, R&D measures the expenditure on research and development programs (% of GDP), and FDI represents the net inflow of foreign direct investment to the investigated region. Similarly, access to clean energy and technology is the percentage of total population, trade ratio is taken in the share of GDP, and population is taken as how many people living per square kilometer of land area.We targeted G-seven economies (developed) over the period of 2001 to 2018 (18 years). According to Global Footprint Network (GFN), these countries are in the list of top 10 countries with high EcoFP . The data for EcoFP are obtained from the official website of GFN (2021) database. As a dependent variable, per capita EcoFP was used as a proxy for environmental sustainability. Regrading independent variables of our models, we used as many as possible, as well as control variables see . We usedThe current study analyzes the relationship between ICT, ECI, and EcoFP, i.e., whether and how ICT reduces ecological pollution in G-seven countries and what is the possible role of economic complexity in this process. In addition, this study also used some control group factors so that it would not omit any important factor. Moreover, all possible variables used in the regressions are converted to logarithmic form to verify homogeneousness in our series. Following the study objective, our EcoFP was model as follows:y represents dependent and X represents independent variables (including main and control variables). In addition, \u2018i\u2019 and \u2018t\u2019 denote cross-sections (countries) and time (years), correspondingly, while \u03ad represents stochastic error term.In the above equation, By considering these variable, our study further proceeds for the specification of the model following Khan et al. (2022), and we present the specification of the model with log relationship as follows (see Equation (2)):Since all variables (dependent and independent variables) in Equation (2) are presented in log form, the estimated parameters in log form represent long-run responsiveness (long-run elasticities).Regarding the estimation process, firstly, the current study investigates the under-examined panel data of G-seven economies for the cross-sectional dependency, which is a very common matter in panel data. To this end, in this study, we have applied Pesaran, et al. CD test i is different cross-sections (countries), t is specific time periods (years), it\u03be and \u03bci represent the deterministic component, while eit expresses the process of stationary series. Regarding second-generation (CIPS) panel unit root testing approach, this paper also suggests a simple stationary test in the occurrence of cross-sectional dependency.In Equations (3) and (4), The existing of cointegrational association among variables can be explored by using Kao panel cointegration and Engle\u2013Granger-based Pedroni test. Based on these tests, the long-run relationship confirms the existence of cointegration; therefore, we applied FM-OLS techniques. Equation (6) provides the general form of the model:In the above equation:ity and itx represent dependent and independent variables; moreover, with partition covariance, \u0439 is the triangle of lower-case matrix. Similarly, asymptotic covariance matrix for cointegration analysis is denoted by \u03a9, while \u0403 presents dynamic covariance in the above equations. Furthermore, in the analysis of current study, we also developed the pooled mean group-ARDL (PMG-ARDL) model that allows for the estimation of both short- and long-run coefficients. In the existing literature, many studies suggest several advantages of PMG-ARDL model over FM-OLS model that can be traced by [Thus, raced by ,65,66.After detection of the cointegration between the dependent and independent variables, the next phase is to estimate the degree of the long-run elasticities of the variables used in the model. To this end, we use FM-OLS methodology to calculate the long-run coefficients. The FM-OLS methodology has the capability to tackle the issue of serial correlation and the problem of endogeneity in the analysis of panel data. FM-OLS suggests a nonparametric approach and has to work in a small sample dataset . Furthermore, we employ another econometric approach to approximate a dynamic nonstationary panel using PMG methodology introduced by Pesaran, et al. . The maiexp is rejected, even at 10%, 5%, and 1% levels. Particularly, lnICTimp, lnGDPpc, lnECI, lnFDI, lnPOP, lnTRA, lnPOP, lnINF, and lnRD are negatively skewed, while the remaining variables (such as lnEcoFP and lnICTexp) obtain positive skewness and, therefore, heavy-tailed distribution.exp, ICTimp, GDP, ECI, and EcoFP . As discussed above, we use the two latest econometric panel techniques, namely \u201cFM-OLS\u201d and \u201cPMG-ARDL\u201d, to actually reflect the net effect of all independent variables on the dependent variable.The phenomenon of heavy-tailed distribution also leads us to apply the most valid and suitable panel techniques to empirically observe the association between ICTSimilarly, in In order to check the stationarity level and order of integration of all series, we utilized IPS and CIPS panel unit root tests, as shown in On the basis of unit root tests, this study further proceeds to scrutinize the long-run linkages between dependent and independent variables used in our regression models. To this end, two cointegration tests have been applied, namely \u201cPedroni\u201d and \u201cKao\u201d cointegration tests. exp), research and development (lnRD), trade (lnTRA), and population (lnPOP) suggest that an upsurge in these factors reduce environmental degradation in terms of ecological footprint (lnEcoFP) in G-seven economies. On the other hand, the statistically significant and positive coefficients of ICT imports (lnICTimp), GDP per capita (lnGDPpc), and economic complexity (ECI) imply that an increase in these factors increase EcoFP in investigated regions. Precisely, the long-run responsiveness (elasticities) of ICT export, ICT import, GDP per capita, economic complexity, research and development, trade, and population are 0.091%, \u22120.079%, 0.193%, 0.071, \u22120.771%, \u22120.092%, and \u22122.044%, respectively . Furthermore, From export and its expansion might not be detected as a reasonable clarification in handling environmental issues [Frist, the coefficient of ICT export is positively and statistically significant with Eco-FP. On average, a 0.232% (FM-OLS I), 0.260% (FM-OLS II), and 0.091% (FM-OLS III) increase in EcoFP is caused by ICT export in the long run. It is worth mentioning that producing extra ICT-related heavy machines, equipment, and infrastructure and exporting of these goods are unfriendly to environmental quality, indicating that it is increasingly EcoFP. Therefore, our study suggests that ICTl issues . These el issues for 21 Sl issues for G-seexport and ICTimport (positive and negative) in the long run. Therefore, all countries from the G-seven region should plan and formulate their policies related to ICT according to the prevailing domestic economic situation. In this regard, the G-seven region considerately needs to increase private\u2013public awareness about environmental issues by utilizing extra resources in their ICT sectors.On the other hand, ICT import is statistically significant and negatively affects EcoFP in the long run. Precisely, a 0.162% (FM-OLS 1), 0.046% (FM-OLS II), and 0.079% (FM-OLS) decrease in EcoFP is caused by ICT import in the long run. These specific empirical results can be defendable by understanding the contributing role of the SE that is positively associated with environmental sustainability in terms of dematerialization, demobilization, and decarbonization activities. The substantial improvement in the ICT sector, especially in ICT import, encourages the implicit activities, for example, online business and academic conferences, online businesses, electronic books, e-commerce, and e-banking, decreases the aggregate demand for nonrenewable energy, and, therefore, decreases environmental unsustainability. The enhancement of SE in mitigating EcoFP has previously ignored the effect of consumption and cost effect. The empirical findings of this study are in line with the work of many researchers, who suggest that the ICT-import-based SE contributes to reducing emission levels by reducing the aggregate demand for energy consumption, encouraging resource competence, and technological development in the ICT sector ,69. The Regarding economic complexity (ECI), a 0.271% (FM-OLS I), 0.273% (FM-OLS), and 0.071% increase in EcoFP is caused by ECI in G-seven economies for the long run. These empirical results of ECI are in line with work of Khan et al. (2022) for the G-seven region, Neagu and Teodoru for EU eWhen it comes to economic growth, the results suggest that economic growth (lnGDPpc) significantly contributes to environmental degradation, with a 0.130% (FM-OLS I), 0.132% (FM-OLS II), and 0.193% (FM-OLS III) increase in EcoFP caused by GDPpc growth or economic growth. The coefficients of GDPpc are statistically significant in all models , as shown in Expenditures on research and development (RD) suggest a significantly negative effect on EcoFP in G-seven economies for the long run. In In the case of control variables, foreign direct investment (FDI), and all models of In this study, we critically analyzed existing papers in the environmental economics literature, in which many researchers used carbon emissions as a proxy for environmental unsustainability, which cannot reflect the major parts of the ecosystem. By taking this into account, in this study, we analyze the linkages between information, communications and technology, economic complexity, and ecological footprint are producing more emissions, which put pressure on the environmental quality. Using different econometric methods, such as panel unit root/stationarity tests (both first and second-generation), panel cointegration tests (Pedroni and Kao), and panel regression analysis (FM-OLS and PMG-ARDL), the empirical evidence obtained from this study concludes that ICT export increases the EcoFP in the long run for G-seven economies, while ICT import contributes to reducing EcoFP in these regions. Furthermore, the empirical findings conclude that economic complexity increases environmental sustainability and plays a more detrimental role to increase EcoFP in all cross-sections. However, the elasticities of coefficients are divers for all regression models used in this study. In addition, economic growth or per capita GDP also play a harmful role by reducing environmental sustainability, whereas FDI does not contribute to affecting EcoFP in the long run. As expected, research and development significantly reduce ecological destructions by spending on environmental-related research and development programs. On the other hand, trade plays a vital role to reduce environmental pollution in the long run, whereas population played a very destructive role to degrade environmental quality in the G-seven region.I.ICT export and ICT import positively and negatively influence environmental sustainability in terms of ecological footprint, respectively. This empirical finding implies that importing highly advanced ICT infrastructure can help to decrease environmental unsustainability in the investigated region. Meanwhile, the production and export of these (ICT infrastructure) might be paid as high environmental cost. This evidence suggests that the governments and policymakers of these regions should implement policy to encourage the penetration of the ICT sector to maintain environmental sustainability.II.The significance of ECI in boosting the economy has been widely considered and accepted in this role. However, current debates on controlling effects of ECI to environmental aspects also draw enormous attention from a wide range of policymakers and concerned authorities. By the same token, the empirical results for economic complexity positively affect EcoFP. This infers that prevailing economic transformation (to knowledge- and skill-based) of industrial structure and economic activities in selected G-seven economies exploits environmental sustainability and is not environmentally friendly. Therefore, the concerned authorities should consider economic activities and complex structure of industries while implementing environmental sustainability policies.III.Likewise, the empirical evidence provides that FDI and trade activities will not help G-seven economies to reduce environmental unsustainability by lowering ecological footprint. Foreign investors and trade are not helpful to bringing environmentally friendly technology to the host countries to reduce environmental unsustainability. Therefore, the governments of the investigated region should encourage foreign investors and domestic traders to bring greener technology, which will not only help the G-seven countries in environmental terms, but also in economic condition.IV.Regarding research and development, it also negatively affects environmental unsustainability by reducing ecological footprint. The governments of the investigated region should invest more in research and development to reduce the harmful effect of economic growth on the environment. The research and development can also be helpful in structure transformation of the economy toward green economy.The specific policy implications and discoveries can be summarized from the empirical findings of this study as follows:In order to strengthen the existing qualitative and quantitative information, the research gap can be highlighted in light of the following caveats and limitations. First, the current study\u2019s emphasis is on the relationship between ICT, ECI, and EcoFP; therefore, it would be of more robust and interesting if future researchers inspect the relationship between ICT and EcoFP on a micro or macro level , specifically, analyzing the relationship between ICT and environmental sustainability in different firms and industries in the G-seven countries. Second, another econometric method should be used to estimate whether the empirical findings of this study are in line with the empirical examination within the homogenous (standardized) panel and country-specific analysis.The current study also leaves the gap for future researchers to fill by investigating developing or emerging economies. The investigation of the current study considered only developed economies; therefore, the findings for developing or emerging economy regions from Asia or sub-Saharan Africa must be different. Thus, future research would improve policy effect and explore many research objectives not even in developed but in the developing world. Moreover, in the analysis of this study, we could not find the short-run elasticities and causality among the variables because of inaccessibility to an extended dataset. It would be a more valid and reliable outcome to handle this constraint in future research. Lastly, this study could not perform the causality and empirical mechanism tests between variables used in the models (due to unavailability of a longer dataset for longer panel). Therefore, it would be better to cover this gap in the future study by conducting empirical mechanism tests to verify the impact mechanism and causality between variables."} +{"text": "The first Uruguay's Report Card in 2018 based on the Global Matrix initiative showed the lack of information on physical activity in children and adolescents. This study mapped and examined the available evidence on physical activity-related indicators based on Uruguay's 2022 Report Card.The scoping review was reported using the Joanna Briggs Institute and the Preferred Reporting Items for Systematic Reviews extension for Scoping Reviews guidelines. A comprehensive literature search was performed for the period between 2018 and 2021, including electronic databases , gray literature , national and regional relevant journals, and reference lists of key texts. Two researchers independently conducted both the selection and data-charting process. Data items from each paper were charted based on the Population, Concept, and Context elements reflected in the objective of the review. A narrative synthesis and network plots were conducted to summarize the evidence.A total of 20 papers were included in this review, consisting of four peer-reviewed scientific papers, three bachelor's theses, four official documents of State-agencies, four Government reports, of which three included national surveys, and five laws. Strengths, weaknesses, and knowledge gaps were identified from the available evidence. We synthesized main challenges such as publishing scientific studies, establishing cross-national and cross-sectoral collaborations in research projects, generating high-quality data, reporting information on social inequality indicators that influence equitable distribution, or increasing access to public information. Our results support early emerging and growth research on this topic. However, despite existing papers on physical activity-related indicators in Uruguayan youths, the lack of high-quality evidence remains clear.The findings of this scoping review provide the best available evidence for identifying and overcoming the challenges of physical activity-related indicators research in Uruguay. The methodological framework used could be useful for countries involved in future editions of the Global Matrix initiative.https://osf.io/hstbd/.Open Science Framework, Physical activity (PA) is a fundamental pillar in the health and well-being of children and adolescents , 2. In cOne of the strategic actions recognized by the World Health Organization (WHO) for this challenge is the continuous improvement of nationwide data systems that support regular PA surveillance . In thisn = 7) to draw consistent conclusions, and the lack of information in this field with three out of 10 indicators with insufficient information was conducted using Arksey and O'Malley's five-stage framework and the An ScR allows for a broad and comprehensive review of the existing literature, and to identify research gaps . TherefoFor the purposes of this ScR, the papers referred to all types of documents, such as scientific publications, government documents or reports, open access theses, and laws. To be included, papers needed to focus on the PCC elements:\u2013 Population: Uruguayan children (5\u201312 years) and adolescents (13\u201317 years).\u2013 Concept: PA-related indicators of the GM initiative linked to daily behaviors , settings and sources of influence , and strategies and investments [government (GOV)].\u2013 Context: Uruguay.We included papers that reported quantitative information on the benchmarks of PA-related indicators in UruguTo identify potentially relevant documents, a comprehensive literature search was performed using two methods.First, a systematic search of peer-reviewed journal papers was conducted on MEDLINE (PubMed), Web of Science, LILACS, Scielo, and Dialnet (Latindex) from 01/012018 up to 31/12/2021. The search strategy was performed using the following terms based on the PCC mnemonic (via Google Scholar (using the main terms based on the PCC mnemonics detailed above), open access thesis , and relevant websites of State-agencies and International Organizations were searched for the identification of papers via other methods.Second, gray literature identified All searches were performed between 2018 and 2021, based on the 4-year update of the literature available for the GM 4.0 project's PA-related indicators gradings reporting . Even soThe screening process was performed by ordering the references using the Mendeley reference management software (version 1.19.8) according to the inclusion/exclusion criteria. An Excel standardized table (v.11) was used for the selection and extraction process to establish agreement among reviewers.First, two independent reviewers (BBP and JBS) screened the titles and abstracts of documents for potential inclusion. In cases where a decision for exclusion or potential inclusion cannot be made, the full text was retrieved. Second, two independent reviewers (BBP and JBS) decided on the inclusion or exclusion of the full-text documents based on the selected criteria by completing a checklist form. We resolved disagreements on study selection by consensus with all reviewers if needed.Two reviewers (BBP and JBS) developed an Excel (v.11) standardized table to chart the data from specific domains of the PCC elements . The folWe contacted one paper's authors to access additional relevant material . Springer et al. sent theAccording to the guidelines for ScR, no quality assessment is required , 22 and,A total of 1,153 records were assessed, comprising 996 studies from databases and 157 reports from websites . Finally, 20 papers were included in this ScR .Of the peer-reviewed scientific papers, three were cross-sectional \u201333 and oThe papers were conducted between 2018 and 2021, except for one official document of a State educational agency and threData samples were collected between 2015 and 2020. Of the papers included (12 out of 20) involving children and adolescents, the range of sample sizes was 55\u2013136,483 , 41\u201343. The most common socioecological setting targeted was the Primary school in 10 papers , 41\u201343, Findings from the included papers are synthesized in The main purpose of the papers included was PA-related indicators, except for two laws that focused on a broader topic such as the education system , 27, twoCompared to the 2018 Report Card (evidence searches from inception to 2018 included seven papers to report the indicator grades), the Uruguay 2022 Report Card (evidence searches between 2018 and 2021) was based on a larger number of papers (19 out of 20 included in this ScR).The strengths of the evidence from scientific publications were indexed in international databases , 33, ranvia other methods, the strengths of the evidence were nationally representative data with updated data on the PA-related indicators were found compared to Uruguay's 2018 Report Card (n = 8) . Althoug (n = 8) .Considering the scientific papers included, our review revealed the first scientific papers indexed in international databases that analyze PA-related indicators in Uruguayan childhood or adolescence , 33. ThiMoreover, two national interdisciplinary research centers , 33 and Considering papers other than scientific publications, our results indicate a major strength with three nationally representative surveys that were conducted periodically , 34, 35.Regarding the PA-related indicators, our review displayed a nationally approached framework for SP (federated sports) and SCH standards. Uruguay's National Sport Plan 2015\u20132020 and the Beyond these advances, our ScR revealed that papers examining the PA-related indicators in Uruguayan children and adolescents remain lacking and high-quality research production is a major challenge. Indeed, nearly half of the indicators in Uruguay's 2022 Report Card were graThe few scientific productions, the small number of cross-national collaborations and research centers, the limited data , and the near absence of papers published on some PA-related indicators display a void in the research capacity concerning this topic in Uruguay. Some weaknesses reported in the Latin American context , 58 coulResults of our ScR suggest that there are many areas to contemplate in future PA research. Specifically, social inequalities remain among the biggest challenges for global PA promotion and shouThe authors recommend that national efforts on PA-related indicators research should be made to address two interrelated priority paths.On the one hand, related to weaknesses in the published literature, they should include scientific publications, specifically with nationally representative data and longitudinal analysis; cross-national and cross-sectoral collaborations in research projects; creation and consolidation of national and transdisciplinary research centers, mostly in cities that have been underrepresented compared to the Uruguayan capital; and national surveys with follow-up data.On the other hand, related to knowledge gaps in the PA-related indicators, they should include: appropriate data, particularly on AP, COM, FAM, PF, and SP; information by gender, particularly on AP, SCH, and GOV; evidence by influence factors ; accessible public information, principally on GOV and SCH ; data by specific children ages, mainly on AT, PA, and SB; knowledge about a whole-school approach that includes components such as modified school policies to engage students with low PA levels or parental engagement; findings on SP in community environments; evidence on public spaces and infrastructure; and device-based PA data.More specifically, the authors recommend that future research initiatives on PA-related indicators should incorporate the methodological GM framework and report data on this basis. This would help improve epidemiological characterization and, therefore, guide policy development to increase PA levels. We also encourage future research projects to have a comprehensive approach that assesses relevant PA-related factors of influence, such as gender or socioeconomic status, for a better reflection of the local context. This information will also be key to developing efficient policies encouraging PA. Finally, the authors call for a synergistic approach to future research proposals that will allow further progress in an emerging and challenging scientific field in Uruguay. Specifically, research strategies emphasizing multicomponent , cross-sectorial , and cross-national initiatives will be key to increasing the level of maturity of PA research on children and adolescents and to guide effective policy actions based on high-quality evidence.This study is the first to provide an ScR on PA-related indicators in children and adolescents from Uruguay. The methodological framework used could be useful for countries with a lack of research on this topic and for new countries in future editions of the GM initiative. Regarding the Uruguayan context, the above-mentioned strengths in PA-related indicators research and evidence are crucial advances to improve research capacity and guide effective policy actions.Our study has limitations that should be acknowledged. Because we conducted an ScR, it is not necessary to rate the quality of the data or conduct a critical appraisal of the evidence included. Furthermore, although we used a broad search strategy, it is possible that some papers were missed. Additionally, no qualitative papers or papers reporting data on specific groups of interest were included, which could exclude relevant data when mapping Uruguay's PA research capacity. We intended to provide an overall picture of the state of the PA-related indicators research in Uruguay based on the GM initiative.This is the first ScR generating a comprehensive view of the physical activity evidence in children and adolescents from Uruguay. Given the lack of previous papers at the national level, our findings provide the best available evidence for identifying and overcoming the challenges of physical activity-related indicators research. Understanding strengths, weaknesses, and research gaps are essential to improve research capacity in this health behavior field. Although papers examining physical activity-related indicators in Uruguayan children and adolescents remain lacking, we displayed some advances in research production. From the public health perspective, the double strength is clear: by improving research capacity, high-quality evidence becomes available to guide effective policy action in Uruguay. Knowing the epidemiological reality should be the main objective in future research to identify evidence-based challenges and priorities for action on physical activity-related indicators. The methodological framework applied could be useful for countries of the Global Matrix initiative.BB-P and JB-S: conceptualization, methodology, formal analysis, investigation, data curation, writing\u2014original draft, writing\u2014review and editing, and visualization. JB-S: supervision. CAC, EP-T, SF-G, and VD-G: data curation and writing\u2014review and editing. All authors contributed to the article and approved the submitted version.Processing Charges have been covered by the PDU EFISAL\u2014Universidad de la Rep\u00fablica (Exp. n\u00b0 003051-000603-16).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "In 2012, the World Health Organization (WHO) released the Global Plan for Insecticide Resistance Management in malaria vectors to stress the need to address insecticide resistance. In a prospective multi-centric study commissioned by the Indian Council of Medical Research (ICMR), we assessed the insecticide susceptibility status of the primary malaria vectors in India from 2017 through 2019.An. culicifacies, An. fluviatilis, An. stephensi, An. minimus and An. baimaii and secondary malaria vectors - An. aconitus, An. annularis and An. philippinensis/nivepes from 328 villages in 79 districts of 15 states of India were assessed following the WHO method mainly to insecticides used in vector control, organochlorine (DDT), organophosphate , and other pyrethroids . The study sites were selected as suggested by the National Vector Borne Disease Control Programme.The insecticide susceptibility status of the prevalent primary malaria vectors \u2013 An. culicifacies showed resistance to DDT (50/50 districts including two districts of Northeastern India), malathion (27/44 districts), and deltamethrin (17/44 districts). This species was resistant to DDT alone in 19 districts, double resistant to DDT-malathion in 16 districts, double resistant to DDT-deltamethrin in 6 districts, and triple resistant to DDT-malathion-deltamethrin in 9 districts. An. minimus and An. baimaii were susceptible in Northeastern India while An. fluviatilis and the secondary malaria vector An. annularis was resistant to DDT in Jharkhand.The primary malaria vector An. culicifacies is predominantly resistant to multiple insecticides. Our data suggest that periodic monitoring of insecticide susceptibility is vital. The national malaria program can take proactive steps for insecticide resistance management to continue its push toward malaria elimination in India.In this study we report that among the primary vectors Anopheles; India; Insecticide-resistance; Malaria vectors."} +{"text": "Brain Sciences, eight topical papers on the topic are presented. The focus of this Special Issue was on CP diagnosis and intervention in both low- and high-resource settings.Cerebral palsy (CP) is the most common physical disability in childhood. Early detection and early intervention for cerebral palsy is now well-proven, evidence-backed, and slowly gathering momentum in hospitals and clinics around the world . In thisTwo papers feature work on the early detection of CP from high-resource clinical settings in Australia. Velde et al. described the single-centre implementation of early diagnosis in a CP clinic in NSW, Australia . They shFive papers in this Special Issue focused on CP diagnosis and management in low- and middle-income settings. King et al. reviewed the literature on early diagnoses of CP in LMICs, highlighting the paucity of evidence and published studies from LMICs in this field . Mushta In summary, the early diagnosis and early intervention of CP is critical for improving the long-term functional outcomes of CP children and their families. This Special Issue showcases some of the work which is being carried out in this field. Most importantly, it further highlights the work needed in LMIC settings to improve understanding of epidemiology of CP, and access to early diagnosis and intervention for these communities."} +{"text": "Visceral leishmaniasis (VL) is a serious vector-borne disease in central and western China. In recent years, the number of VL cases increased gradually, particularly the mountain-type zoonotic visceral leishmaniasis (MT-ZVL). This study clarified the epidemiological features and spatial-temporal clustering of VL in China between 2019 and 2021, identified the risk areas for VL transmission, and provided scientific evidence for the prevention and control of VL.Moran\u2019s I and Getis-ORD Gi* statistical data were processed for spatial autocorrelation and hotspot analysis in ESRI ArcGIS software. Also, spatial-temporal clustering analysis was conducted with the retrospective space\u2013time permutation scan statistics.The information on VL cases in 2019\u20132021 was collected from the Infectious Disease Reporting Information Management System of the Chinese Center for Disease Control and Prevention. The epidemiological characteristics of VL cases were analyzed. The global A total of 608 VL cases were reported from 2019 to 2021, with 158, 213, and 237 cases reported each year, respectively. Of the 608 cases, there were 10 cases of anthroponotic visceral leishmaniasis (AVL), 20 cases of desert-type zoonotic visceral leishmaniasis (DT-ZVL), and 578 cases of MT-ZVL. The age of VL cases was mainly distributed in the group of subjects aged \u2265 15 years. Peasants and infants were the dominant high-risk population. The incidence peak season of VL occurred between March and May. The cases were mainly distributed in Shanxi (299 cases), Shaanxi (118 cases), and Gansu (106 cases) Provinces, accounting for 86.02% (523/608) of the total reported cases in China. Spatial analysis revealed that clustering of infection is mainly located in eastern Shanxi Province and Shaanxi\u2013Shanxi border areas, as well as southern Gansu and northern Sichuan Province. In addition, new reemergence hotspots in Shanxi, Henan, and Hebei Provinces have been detected since 2020. Spatio-temporal clustering analysis revealed an increase in the degree of infection aggregation in eastern Shanxi Province and Shaanxi\u2013Shanxi border areas.The AVL and DT-ZVL were endemic at a lower level in western China, whereas MT-ZVL rebounded rapidly and showed a resurgence in historically endemic counties. The spatial-temporal clustering analysis displayed that the high-incidence areas of VL have shifted to central China, particularly in Shanxi and Shaanxi Provinces. Integrated mitigation strategies targeting high-risk populations are needed to control VL transmission in high-risk areas. Visceral leishmaniasis (VL), also known as Kala-azar, is a serious infectious disease caused by Leishmania spp. that is transmitted by the bite of phlebotomine sand flies , the hosIn the early 1950s, VL was one of the five most serious parasitic diseases endangering human health in the People\u2019s Republic of China. It was prevalent in 16 provinces located north of the Yangtze River, with about 530,000 patients . In 1956Leishmania donovani was the vector and dogs were the reservoirs with high infection rates. MT-ZVL was hypoendemic in humans and affected mainly infants under 5 years of age. The incidence rate in humans is much lower than the rate in canines, and no obvious correlation was found between the cases was the primary vector . The incThese three types of VL exhibit substantial differences in epidemiological characteristics, i.e., geographical and landscape features, ecosystem, vector species, and infectious sources , and theIn recent years, a rapid reemergence of VL has been seen in counties that had previously achieved elimination in China. It presented a significant challenge for the control of the disease. From the year 2019, public health agencies have been recruited by the government to perform epidemiological investigations on each VL case, to trace it back to their original infection place. This detailed information will facilitate more accurate risk identification than before, and analysis of epidemiological features and spatial-temporal clustering of VL.Scholars have carried out some studies on the spatial-temporal clustering of VL. In this study, large-scale epidemiological investigations on each case were conducted for the first time to determine their specific infected counties. We classified VL cases into MT-ZVL, AVL, and DT-ZVL, and elucidated their epidemiological features and spatial-temporal clustering of all three different epidemic types of VL, which will be helpful for guiding the accurate prevention and control of VL.In 2004, the web-based National Diseases Reporting Information System (NDRIS) was established in China. It is a network covering all the medical and health institutions at or above the township level. According to the Law of the Infectious Disease Control and Prevention, each medical and health institution in the country has to notify the cases of VL online within 24 h after the clinical cases are confirmed with the diagnostic criteria of VL in China (WS258-2006). Then the county CDC, where the cases are reported, will take responsibility to conduct the epidemiological investigation of individual cases in 7 days to determine whether the patient is an autochthonic case or not, and subsequently take countermeasures onsite according to relevant guidance. The information of the cases reported through NDRIS includes age, gender, occupation, current residential address, symptom onset date, diagnosis date, and therapeutic outcome, based on which demographic distribution, geographic distribution, temporal distribution, and the lag time between symptom onset and diagnosis can be analyzed.In this study, VL case data, reported in mainland China from 2019 to 2021, were obtained from the National Notifiable Communicable Disease Reporting System . Suspectt-test was used to judge the significant difference in the incidence trend of VL during a certain period used mathematical statistical analysis to search for turning points and divided the trend of disease incidence over a long period into several statistically significant sections for analysis. In this study, the number of VL cases in 2015\u20132018 was obtained from the National Notifiable Communicable Disease Reporting System . We analn period . The lonn period . The \u201ccrSpatial autocorrelation is defined as the correlation between values of a single variable at different geographical locations using a measurement of spatial clustering based on feature locations and attribute values , and theI is indicative of the Moran\u2019s I statistic, with values ranging from -1 (perfect dispersion) to 1 (perfect correlation). Negative values indicate dispersion, positive values mean clustering, and a value of zero indicates no spatial correlation or low values (cold spot) with statistical significance judged by P-values . It is cGi* statistics is an indicator of local aggregation. If the value of Gi* is greater than 0, it indicates that the neighbor attribute value of its spatial unit i is high; otherwise, the neighbor attribute value is low. Local spatial autocorrelation at the county level is reflected by the local indicators of spatial association (LISA). The LISA clustering map involves four patterns: high-high, high-low, low-high, and low-low.p-value. A p-value < 0.05 was indicative of a statistically significant cluster. The grade is based on the value of the log-likelihood ratio (LLR) to generate Grade I, Grade II, Grade III, and Grade IV cluster areas. According to the P-value and LLR value, it is determined whether there is clustering in the study area, and its exact location and risk size are also analyzed. Finally, the result was visualized through ESRI ArcGIS software (version 10.3).Spatio-temporal statistics were employed to describe the temporal and spatial distribution, and to detect the geographic and temporal clusters of VL during 2019\u20132021. A Poisson model using a retrospective space\u2013time permutation scan statistic was used to identify spatio-temporal clusters of VL using SatScan software version 9.4.2 . Space\u2013tBetween 2019 and 2021, a total of 612 VL cases were reported in China, including one fatality. The annual incidence increased year by year, with 161, 214, and 237 cases reported each year, respectively. After the epidemiological investigation of individual cases, the origin of 608 cases, including 10 cases of AVL, 20 cases of DT-ZVL, and 578 cases of MT-ZVL, can be traced to the county level. However, the other four cases could not be traced to the source of infection and were not included in the epidemiological analysis .The VL cases were mainly distributed in the age group \u2265 15 years old, accounting for 69.57% (423/608) of the total cases. AVL, DT-ZVL, and MT-ZVL cases were mainly distributed in the age group \u2265 15 years old, accounting for 60.00, 65.00, and 69.90% of the total cases of each type, respectively .In terms of occupation, peasants are the highest risk population for VL (285/608), followed by infants and young children (165/608). For AVL, the predominant cases (4/10) were peasants. For DT-ZVL, most of the cases were peasants (10/20). For MT-ZVL, the majority of cases were also peasants (271/578) followed by infants and young children 156/578; .In addition, the male/female ratio of VL cases in China was 421:191, and the male cases were significantly higher than the female cases. The ratio of males to females was 1:1, 1:0.25, and 1:0.46 for AVL, DT-ZVL, and MT-ZVL, respectively .The incidence peak varied in different types of VL. For MT-ZVL, it appeared in the period from March to May. However, no obvious peak of AVL and DT-ZVL was observed due to the low number of cases .P < 0.05). On the contrary, the incidence of DT-ZVL decreased rapidly , the Shaanxi\u2013Shanxi border areas , eastern Shanxi (Yangquan City), and northern Henan Province (Linzhou City). In addition, a total of 54 VL cases were detected in 24 resurgence counties which were mainly located in Henan (8), Shanxi (6), Hebei (5), Shaanxi (4), and Gansu (3) Provinces, and Beijing Municipality (1) .In the last 3 years, the high-high clusters were mainly observed in the eastern Shanxi (Yangquan), the border areas of Shaanxi\u2013Shanxi , southern Gansu and northern Sichuan , and southeast Shanxi Province (Changzhi). Since 2020, new high-high clusters began to occur in the northwestern Henan Province (Zhengzhou) and southern Xinjiang (Kashi Prefecture). In 2019, high-low clustering appeared in western Inner Mongolia (Ejina Banner) and eastern Xinjiang (Hami City), since sporadic cases were reported in these sparsely populated areas .Consistent with the result of spatial autocorrelation analysis, hotspots were predominantly identified in eastern Shanxi, the border areas of Shaanxi\u2013Shanxi, southern Gansu, and northern Sichuan, as well as southeast Shanxi Province (Changzhi), from 2019 to 2021. In addition, new hotspots were identified in northwestern Henan Province and southern Xinjiang from 2020 to 2021 .p-value < 0.05 was indicative of a statistically significant cluster.The result, using LLR to estimate the degree of infection clustering in the incidence of MT-ZVL in China from 2015 to 2021, most notably in Shanxi and Shaanxi Provinces, revealing an observable increase in the incidence of MT-ZVL in these areas. On the contrary, the incidence of DT-ZVL decreased rapidly . It may be due to the large-scale renovations of old housing constructions supported by poverty alleviation campaigns in rural areas, particularly in southern Xinjiang which was the most affected area in China. The greatly improved housing conditions of residents destroyed the breeding sites of peridomestic sand flies, and reduced the frequency of contact between the population and sand flies , Shaanxi (19.28%), and Gansu (17.32%), rather than Xinjiang Uygur Autonomous Region which was the predominant former endemic area. It indicates that the main endemic areas shifted from the Xinjiang Uygur Autonomous Region in western China to Shanxi, Shaanxi, and Gansu Province in central China . During nd flies .Three types of VL exhibit substantially different epidemiological characteristics in China . AVL is The substantial increase in the incidence of MT-ZVL in China may be related to the emergence of VL in the historically endemic areas and the formation of some new hotspot areas of MT-ZVL . BetweenPhlebotomus sinensis(Ph. sinensis), to the extent that the density of Ph. sinensis in Yangquan City has reached as high as 103 sandflies/per person per hour recently , particularly for the high-risk region (Grade I), revealing spatial clustering of VL in the whole country for each year of the study period. In the past 3 years, the high-incidence clusters were maintained mainly in the eastern and southeast Shanxi Province, the border of Shanxi Province, and Shaanxi Province, as well as the border of southern Gansu and northern Sichuan regions, demonstrating that although southern Gansu and northern Sichuan were still traditionally the high-incidence regions, two new higher-incidence regions have formed in Yangquan City and Hancheng City in recent years, where they have become the most predominant endemic areas of VL in China. The spatio-temporal cluster analysis also showed similar results that the clusters located in eastern Shanxi Province (Yangquan City) and Shaanxi\u2013Shanxi border areas have surpassed those located in southern Gansu and northern Sichuan, and became the top grade clusters. These areas are extension regions of the Loess Plateau and are mainly hilly settings, where the Yanshan-Taihangshan mountain deciduous broad-leaved forest ecological zone and Fenwei Basin Agro-ecological zone are located . The hilrecently . At the recently ,b. In coSince 2020, small clusters located in southern Xinjiang indicated that risks of DT-ZVL still existed in the desert areas of southern Xinjiang, so we still need to pay close attention to these areas in case of their cyclical outbreaks. Scattered small clusters that appeared in northwestern Henan, southeast Shanxi, western Hebei, and northern Beijing showed that MT-ZVL is reemerging in the Yanshan-Taihangshan mountain regions. Therefore, there is an urgent need for better cooperation among multi-sectors of government for the prevention and control of further extension of MT-ZVL in the surrounding regions.Ph. sinensis have been detected in the densely populated main city of Yangquan, indicating that there is a trend of urbanization of MT-ZVL, which has been reported in Europe and South America (Several phenomena discovered recently have made the prevention and control of VL more complicated. The first is that autochthonic cases, canine leishmaniasis, and America . The sec America , indicat America . StrictlThis study showed that the number of DT-ZVL and AVL cases decreased, while MT-ZVL cases increased quickly. Therefore, it is urgent to strengthen organizational leadership and systematically carry out necessary interventions to prevent further growth of MT-ZVL cases in hotspots and clustering areas, especially in Shanxi and Shaanxi Provinces.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.YL, ZZ, and SL: conceptualization. YL, ZWL, YH, ZZ, and SL: data curation. YL, ZWL, ZZ, and SL: formal analysis and methodology. YL, ZWL, YH, YZ, ZZ, and SL: funding acquisition. YL, LY, ZQL, and ZZ: investigation. YZ and SL: project administration and supervision. ZWL: software. YL, ZWL, and ZZ: writing of the original draft. ZZ, YZ, and SL: reviewing and editing. All authors contributed to the article and approved the submitted version."} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Dear Editor,Globally, TB remains a leading cause of death by an infectious disease.Mining surveillance data,Zoe App in the United Kingdom [covid.joinzoe.com/data] and the Johns Hopkins University dashboard for COVID-19 [coronavirus.jhu.edu/map]), or the use of digital tools for communication and surveillance in Kerala, India.Technological infrastructure continues to grow rapidly and has the potential to enhance real-time big data generation, storage and analysis. Such infrastructure includes information and communication technology with more speed, flexibility and reliability than traditional data systems .The use of big data to provide more in-depth, high-quality insights on the social determinants of TB, as part of an integrated approach to improve healthcare delivery, could help in the identification of the \u201cmissing millions\u201d with TB. Despite its promise, to date, the use of big data in the field of TB has been limited to describing the global burden,"} +{"text": "Wealthy nations must step up support for Africa and vulnerable countries in addressing past, present and future impacts of climate changeThe 2022 report of the Intergovernmental Panel on Climate Change paints a dark picture of the future of life on earth, characterised by ecosystem collapse, species extinction and climate hazards such as heatwaves and floods.While the Paris Agreement of 2015 outlines a global action framework that incorporates providing climate finance to low-income and middle-income countries, this support has yet to materialise.The climate crisis has had an impact on the environmental and social determinants of health across Africa, leading to devastating health effects.Droughts in sub-Saharan Africa have tripled between 1970\u20131979 and 2010\u20132019.The damage to Africa should be of supreme concern to all nations. This is partly for moral reasons. It is highly unjust that the most impacted nations have contributed the least to global cumulative emissions, which are driving the climate crisis and its increasingly severe effects. North America and Europe have contributed 62% of carbon dioxide emissions since the Industrial Revolution, whereas Africa has contributed only 3%.Yet it is not just for moral reasons that all nations should be concerned for Africa. The acute and chronic impacts of the climate crisis create problems like poverty, infectious disease, forced migration and conflict that spread through globalised systems.The primary focus of climate summits remains to rapidly reduce emissions so that global temperature rises are kept to below 1.5\u00b0C. This will limit the harm. But, for Africa and other vulnerable regions, this harm is already severe. Achieving the promised target of providing US$100 billion of climate finance a year is now globally critical if we are to forestall the systemic risks of leaving societies in crisis. This can be done by ensuring these resources focus on increasing resilience to the existing and inevitable future impacts of the climate crisis, as well as on supporting vulnerable nations to reduce their greenhouse gas emissions: a parity of esteem between adaptation and mitigation. These resources should come through grants not loans, and be urgently scaled up before the current review period of 2025. They must put health system resilience at the forefront, as the compounding crises caused by the climate crisis often manifest in acute health problems. Financing adaptation will be more cost-effective than relying on disaster relief.Some progress has been made on adaptation in Africa and around the world, including early warning systems and infrastructure to defend against extremes. But frontline nations are not compensated for impacts from a crisis they did not cause. This is unfair, and drives the spiral of global destabilisation, as nations pour money into responding to disasters, but can no longer afford to pay for greater resilience or to reduce the root problem through emissions reduction. A financing facility for loss and damage must now be introduced, providing additional resources beyond those given for mitigation and adaptation. This must go beyond the failures of COP26, where the suggestion of such a facility was downgraded to \u2018a dialogue\u2019.The climate crisis is a product of global inaction, and comes at great cost to disproportionately impacted African countries, and to the whole world. Africa is united with other frontline regions in urging wealthy nations to finally step up, if for no other reason than that the crises in Africa will sooner rather than later spread and engulf all corners of the globe, by which time it may be too late to effectively respond. If so far they have failed to be persuaded by moral arguments, then hopefully their self-interest will now prevail.East African Medical Journal; Gregory E. Erhabor, Editor-in-Chief, West African Journal of Medicine; Aiah A. Gbakima, Editor-in-Chief, Sierra Leone Journal of Biomedical Research; Abraham Haileamlak, Editor-in-Chief, Ethiopian Journal of Health Sciences; Jean-Marie Kayembe Ntumba, Chief Editor, Annales Africaines de Medecine; James Kigera, Editor-in-Chief, Annals of African Surgery; Laurie Laybourn-Langton, University of Exeter; Bob Mash, Editor-in-Chief, African Journal of Primary Health Care & Family Medicine; Joy Muhia, London School of Medicine and Tropical Hygiene; Fhumulani Mavis Mulaudzi, Editor-in-Chief, Curationis; David Ofori-Adjei, Editor-in-Chief, Ghana Medical Journal; Friday Okonofua, Editor-in-Chief, African Journal of Reproductive Health; Arash Rashidian, Executive Editor, and Maha El-Adawy, Director of Health Promotion, Eastern Mediterranean Health Journal; Siaka Sidib\u00e9, Director of Publication, Mali M\u00e9dical; Abdelmadjid Snouber, Managing Editor, Journal de la Facult\u00e9 de M\u00e9decine d\u2019Oran; James Tumwine, Editor-in-Chief, African Health Sciences; Mohammad Sahar Yassien, Editor-in-Chief, Evidence-Based Nursing Research; Paul Yonga, Managing Editor, East African Medical Journal; Lilia Zakhama, Editor-in-Chief, La Tunisie M\u00e9dicale; Chris Zielinski, University of Winchester.Lukoye Atwoli, Editor-in-Chief, https://www.bmj.com/content/full-list-authors-and-signatories-climate-emergency-editorial-october-2022This comment is being published simultaneously in multiple journals. For the full list of journals, see:"} +{"text": "Addenbrooke\u2019s cognitive examination (ACE) is a cognitive screening tool that has developed through three stages: ACE, ACE-Revised (ACE-R), and ACE-\u2162. In addition, mini-Addenbrooke\u2019s Cognitive Examination (M-ACE) and ACE mobile are the additional versions that is derived from ACE-III. ACE and its related versions show better performance than Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) in detecting mild cognitive impairment in different neurological disorders. It has been translated into numerous languages, including Chinese. Through reviewing the history, validity, and comparison with other cognitive tests of Chinese versions of ACE, it aims to facilitate the clinical and scientific use, further development, improvement, and validation of Chinese versions of ACE in various neurological disorders and ultimately promote early identification and management of cognitive impairment in China. Dementia is a major challenge for global public health . CurrentThis narrative review followed the PRISMA statement. A bibliographic search of studies was conducted until January 2022 using the following key words: \u201cAddenbrooke\u2019s Cognitive Examination, Chinese, China, evaluate, evaluation, validation\u201d. The databases, including PubMed, Google Scholar, and Web of Science, were used for literature searches. Non-English and literature without full-text available were excluded. Articles that used but without evaluation of the Chinese ACEs were excluded. Two reviewers will independently screen the articles. Disagreements between reviewers will be solved by consensus or through the participation of a third reviewer. Following this literature screening standard, nine articles were selected . QualityThe development of ACE has gone through three stages: ACE, Addenbrooke\u2019s Cognitive Examination-Revised (ACE-R), and Addenbrooke\u2019s Cognitive Examination III (ACE-III). ACE was developed by Professor John R Hodges in 2000 . It was The Chinese version of ACE-R, the same as the original version, consists of five cognitive domains: attention/orientation, memory, fluency, language, and visuospatial. It takes about 12 to 20 min to complete the test, and the total score is 100. There are a few modifications based on the underlying principle during the translation process. For example, the name and address in the memory, recall, and recognition section are replaced by Chinese name and address; the letter \u2018P\u2019, which generated as many words as possible, is replaced with Chinese character \u2018che, \u8f66\u2019 in the verbal fluency section; English words and sentences are replaced by Chinese characters or poems with difficulties to produce in the repetition section .The Chinese version of ACE-R is a reliable examination test for detecting cognitive impairment, with its satisfactory sensitivity , specificity , and area under curve (AUC) 0.945, 0.836) to detect mild AD and MCI, respectively 23]. Th. Th23]. , 0.836 tMore clinicians and medical researchers are using Chinese ACE-III in cognitive assessment, as ACE is gradually modified and improved. Thus, there are more studies using the Chinese version of ACE-III than ACE-R 36,43,44,52. ACESimilar to ACE-R, aside from detecting early stages of AD, ACE-III has been used for tracking performances of cognitive domains in various neurological disorders ,33,34,35Due to the wide range of cognitive domains assessed and patients\u2019 cooperation, it usually takes 12 to 20 min to complete the ACE-III test, so the usage of ACE-III may be limited by time constraints in some specific conditions. Thus, M-ACE, a shorter version of ACE-III, was created for this situation in 2015. M-ACE consists of 5 items with a maximum score of 30. The Chinese version of M-ACE is a reliable and quick examination test to detect MCI and mild dementia with its fair sensitivity , specificity , and AUC 37]. Pa. Pa37]. Compared with the MMSE, the Chinese version of ACE-R has a higher sensitivity and AUC to screen for MCI 23], wh, wh23], Unlike the MMSE, which is unidimensional and provides a global deterioration of intellect, ACE-III is multidimensional and can be scored independently according to its five components: attention/orientation, memory, language, verbal fluency (executive functions), and visuospatial skills to generate a cognitive profile .The verbal fluency of ACE-III provides good evaluation value for assessing frontal lobe function. ACE-III shows fewer ceiling effects and better performance in detecting MCI than the MMSE ,52, simiThe Chinese version of M-ACE appears to have a better performance in detecting MCI and mild dementia than the MMSE with higher sensitivity, specificity, and accuracy 37,39].,39.37,39All Chinese versions of ACE-R, ACE-III, and M-ACE have been proven reliable, sensitive, and valid in cognitive screening. In addition, the latest Chinese version of ACE-III and its shorter version, M-ACE, have been proven to be superior to the MMSE and MoCA, the most widely used cognitive screening tools, especially in individuals with higher education. ACE-III has been proven to be superior to ACE-R, and it is more widely used than previous versions in medical research. In addition, previous studies using the Chinese ACE-III for cognitive screening are mostly monocentric studies, and a large multicenter study is required to further validate the efficacy of ACE-III in different geographic regions of China with different dialects. In addition, the Chinese ACE-III has not been widely used in clinics, and the parallel Chinese versions are not fully verified. Furthermore, ACE-III is freely available for medical research, whereas other screening tools may be restricted by copyright (such as MMSE and MoCA). At present, the Chinese version of ACE-III is recommended by the Chinese Medical Association in the white paper \u2018Standardized Protocol for diagnosis and treatment of cognitive impairment\u2019 as a cognitive screening tool in 2021 . CurrentFinally, mobile ACE is a pragmatic tool with its convenience, automation, easy storage, and management. In the clinical usage of paper-based ACE-III, 78% of the usage had either incorrect answers or arithmetical errors, and mobile ACE can reduce the errors by 85\u201393% . TherefoThere are several limitations of this review. We excluded studies applying the Chinese version of ACE without systematic evaluation, as there were no raw data available, which reduced the number of studies reviewed. In addition, there is no electronic version of the Chinese ACE-III, making the Chinese ACE versions incomplete in relation to English versions.To decide which ACE version is conducted, there are several factors to be considered: for research purposes, ACE-III is widely used for comprehensive cognitive assessment, and M-ACE is most likely to be used in busy clinics due to time constraints. Although the latest Chinese version of ACE-III is recommended, it needs to be translated into multi-ethnic language versions and applied in multiple regions, with comparable other cognitive scales to further verify its effectiveness in different neurological disorders. Furthermore, the Chinese electronic version of ACE-III needs to be developed and promoted to be widely used in China. We hope that the wide application of ACE will promote early identification and management of cognitive impairment in China in the future."} +{"text": "Osteonecrosis of the femoral head (ONFH) is a progressive and disabling disease with severe socioeconomic burdens. In the last 30 years, a growing number of publications have reported significant advances in understanding ONFH. However, only a few studies have clarified its global trends and current status. Thus, the purpose of our study was to summarize the global trends and current status in ONFH through bibliometrics.Publications related to ONFH from 1991 to 2020 were searched from the Web of Science (WOS) core collection database. The data were analyzed with bibliometric methods. Microsoft Excel was used for statistical analysis and to draw bar charts. SPSS was applied to perform linear regression analysis. VOSviewer was used to conduct bibliographic coupling analysis, co-authorship analysis, co-citation analysis and co-occurrence analysis.Clinical Orthopaedics and Related Research was the major publishing channels for ONFH-related articles. Takuaki Yamamoto published the most ONFH-related articles. Studies regarding ONFH could be divided into five clusters: 1) mechanism study, 2) treatment study, 3) complication study, 4) radiological study and 5) etiological study. Mechanism study might become a hot spot in the future.A total of 5,523 publications were covered. The United States consistently ranked first in total publications, sum of times cited, average citations per item and H-index. Kyushu University was the main contributor to ONFH. The total number of publications in ONFH has generally increased over the last three decades. The United States was the leading country in ONFH research. Transplantation, engineering, cell and molecular biology, pharmacology and endocrinology have gradually increased and become hot topics in ONFH research. Mechanism study in ONFH including mesenchymal stem cells, apoptosis, oxidative stress, adipogenesis, osteogenic differentiation and endothelial progenitor cells, have attracted more attention and will become a hot spot in the future. Osteonecrosis of the femoral head (ONFH) is a disease in which local death of osteocytes and bone marrow components occurs due to venous stasis or impaired arterial blood supply or disruption of the femoral head . ONFH caIn the last 30 years, a growing number of publications have reported significant advances in the pathogenesis and treatment of ONFH worldwide. However, to the best of our knowledge, only a few studies have clarified the global trends and current status in ONFH. Bibliometrics is a method to cognize the global trends and current status of a certain field and evaluate the contributions of a collection of research results such as all publications of the same scholar, institution or country, by collecting the metrology characteristics of the publications \u201314. In aJournal of Bone and Joint Surgery-British Volume from the 2014 edition and Proceedings of the Institution of Mechanical Engineers. Part H-Journal of Engineering in Medicine from 2002 edition.The search was conducted using the Web of Science (WOS) Core Collection database, including Science Citation Index Expanded (SCI-Expanded), Social Sciences Citation Index (SSCI), Arts and Humanities Citation Index (A&HCI), Conference Proceedings Citation Index-Science (CPCI-S), Conference Proceedings Citation Index-Social Science and Humanities (CPCI-SSH), Book Citation Index-Science (BKCI-S), Book Citation Index-Social Science & Humanities (BKCI-SSH), Emerging Sources Citation Index (ESCI), Current Chemical Reactions Expanded (CCR-Expanded) and Index Chemicus (IC). The journal impact factors (IF) came from the 2020 version Journal Citation Reports except for All the literature was retrieved in WOS on September 2, 2021. The search terms were TS = AND LANGUAGE: (English) AND DOCUMENT TYPES: (Article OR Review). For the time span, we chose the 30 years between 1991 and 2020.Full records and cited references were extracted from the retrieved literature for bibliometric analysis, such as titles, years of publications, authors, nationalities, institutions of authors, funding sources, journals of publications, abstracts, keywords, total number of publications, sum times of cited, average citations per item and H-index. The information based on bibliometric characteristics was downloaded from WOS and imported into Microsoft Excel 2017 and VOSviewer (v.1.6.17) for analysis.P < 0.05 were considered statistically significant. R2 represents the degree to which the linear regression model explains the overall variance was applied to perform linear regression analysis on the trends of total publications in the last 30 years. Any p values variance .VOSviewer is a software for plotting maps based on network data. In the network visualization, items are represented by circles. The size of the circle is determined by the number of publications of the item. The distance between two circles approximately indicates the relatedness of the items. The color of an item is determined by the cluster to which the item belongs. In this study, VOSviewer was used for conducting bibliographic coupling analysis, co-authorship analysis, co-citation analysis and co-occurrence analysis.R2 = 0.872, P < 0.001). The annual publications regarding ONFH have grown nearly sevenfold over the three decades from 61 in 1991 to 481 in 2020 , China , Japan , Germany , England and South Korea was the most active journal in ONFH research, with 348 articles, followed by International Orthopaedics , with 186 articles; Journal of Bone and Joint Surgery-American Volume , with 183 articles; Journal of Arthroplasty , with 154 articles; Journal of Pediatric Orthopaedics , with 132 articles; and Archives of Orthopaedic and Trauma Surgery , with 129 articles. From the top ten journals, we could track the latest advances in ONFH by monitoring the latest research of these journals.The top ten journals publishing the most ONFH-related literature are shown in Clinical Orthopaedics and Related Research published the most ONFH-related articles, nearly twice as many as the publications of the second-ranked journal, International Orthopaedics. Additionally, Clinical Orthopaedics and Related Research had the most total citations, although the average citations per item were relatively low among the top ten journals. In contrast, the third-ranked journal, Journal of Bone and Joint Surgery-American Volume, had the highest average citations and H-index . In addition, Journal of Bone and Joint Surgery-British Volume had relatively high average citations per item and H-index . The higher average citations per item and H-index to some extent indicated the high quality of their publications.Among the top ten journals, The ONFH-related articles published from 1991 to 2020 were categorized into 83 different research areas on WOS. However, to more intuitively show the changes in the research areas over time, we merged the areas of similar significance and finally selected 12 research areas. Orthopaedics, surgery and medical imaging have been extensively and intensively studied due to their focus on the risk factors, etiology, diagnosis and treatment of ONFH. With the development of vascularized bone grafts, bone marrow mesenchymal stem cell transplantation and endothelial progenitor cell transplantation, an increasing number of studies have been conducted to test the efficacy and safety of transplant therapy for ONFH through animal experiments or clinical trials. Furthermore, researchers combined transplantation with cell and molecular biology to enhance the efficacy of transplant therapy for ONFH. For example, Hang et\u00a0al. evaluated the efficacy of vascular endothelial growth factor 165 (VEGF165) transgenic bone marrow mesenchymal stem cells in mongrel dogs with ONFH . In the Bibliographic coupling analysis is a method exhibiting the relatedness of items based on the number of references they share, which is established when two items cited the same article. Bibliographic coupling analysis was used in this study to establish the similarity relationship among publications from the three dimensions of country, institution and journal. To some extent, the total link strength of a particular item can explain its worldwide influence. Items with higher total link strength indicate that the countries/institutions/authors are more globally influential.Clinical Orthopaedics and Related Research , Journal of Bone and Joint Surgery-American Volume , International Orthopaedics , Journal of Arthroplasty , Journal of Bone and Joint Surgery-British Volume and Archives of Orthopaedics and Trauma Surgery . Therefore, Clinical Orthopaedics and Related Research was the leading journal in ONFH globally according to bibliographic coupling analysis, corresponding to the highest total publications and sum of times cited of Clinical Orthopaedics and Related Research in ONFH presented earlier.Co-authorship analysis is a measure to determine the connectivity of items based on the number of co-authored publications. Co-authorship analysis was used to evaluate the cooperation between items in our study by counting the number of co-authored publications. In a way, the total link strength of a particular item can reflect the extent to which they are willing to cooperate with others. Items with higher total link strength indicate that the countries/institutions/authors are more willing to collaborate with others.Co-citation analysis refers to a method presenting the relatedness of items based upon the number of times they are cited together, which is established when two items are both cited in another article. Compared with bibliographic coupling analysis, co-citation analysis can more scientifically highlight the influence of items. Items with higher total link strength indicate that the journals/publications are more influential globally.Clinical Orthopaedics and Related Research , Journal of Bone and Joint Surgery-American Volume , Journal of Bone and Joint Surgery-British Volume , Journal of Arthroplasty , Journal of Pediatric Orthopaedics and Radiology . Therefore, Clinical Orthopaedics and Related Research was the predominant journal in ONFH globally according to co-citation analysis.The co-occurrence network visualization is created by analyzing the number of articles in which keywords occurred together in titles or abstracts. The aim is to determine the hot research directions and topics critical for tracking the development of science . As illuvia Runx2, TAZ, PPAR\u03b3 and C/EBP signaling pathways, which was a pivotal pathogenesis of ONFH , National Natural Science Foundation of China , Hunan Young Talents of Science and Technology (No.2021RC3025) and Elite Medical Professionals Project of China-Japan Friendship Hospital (ZRJY2021-QM21).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Correction: BMC Public Health 22, 1177 (2022)https://doi.org/10.1186/s12889-022-13577-zIn the original publication of this article the author \u2018Kelly Harris\u2019 was accidentally omitted. The original article has beenAuthors\u2019 contributionsSH drafted the manuscript and contributed to the logistical framework of the study. SM contributed to the development and design of the study, data collection tools, IRB approval and continual modifications, and continued quality assurance. SM substantially participated in the drafting protocol and data management. BB contributed to the development and design of the study, data collection, and the maintenance of community partnership. BM helped in solidifying partnerships with the school districts, aided in study design, and data collection. BM participated in drafting the manuscript. NM and KH contributed to the Study design and drafting of the manuscript. SAR, SMR, JP, and JS aided in acquisition of data. In addition to data collection, CE contributed to engaging community partners and maintaining these relationships. TW contributed to data management and analysis, in addition to data collection. SA and GY helped in data collection with a heavy focus in the contract tracing data. ND contributed to the drafting of the manuscript and data analysis. EL participated in the study design and provided expertise on study outcomes and data analysis plan. El participated in drafting the manuscript. AL and IL provided expertise in Informatics and data management. SF aided in designing the trial and drafting the manuscript. JS helped design the clinical trial, identify partnerships and school districts, and drafted and revised the manuscript. All authors reviewed and approved of a final draft."} +{"text": "In light of these findings, the World Health Organization (WHO) recommends that countries implement surveillance strategies to detect the presence of plasmid-mediated colistin-resistant microorganisms and take suitable measures to control and prevent their dissemination. Seven years later, ten different variants of the mcr gene (mcr-1 to mcr-10) have been detected worldwide in bacteria isolated from humans, animals, foods, the environment, and farms. However, the possible transmission mechanisms of the mcr gene among isolates from different geographical origins and sources are largely unknown. This article presents an analysis of whole-genome sequences of Escherichia coli that harbor mcr-1 gene from different origins and geographical location, to identify specific patterns related to virulence genes, plasmid content and antibiotic resistance genes, as well as their phylogeny and their distribution with their origin. In general, E. coli isolates that harbor mcr-1 showed a wide plethora of ARGs. Regarding the plasmid content, the highest concentration of plasmids was found in animal samples. In turn, Asia was the continent that led with the largest diversity and occurrence of these plasmids. Finally, about virulence genes, terC, gad, and traT represent the most frequent virulence genes detected. These findings highlight the relevance of analyzing the environmental settings as an integrative part of the surveillance programs to understand the origins and dissemination of antimicrobial resistance.The importance of the One Health concept in attempting to deal with the increasing levels of multidrug-resistant bacteria in both human and animal health is a challenge for the scientific community, policymakers, and the industry. The discovery of the plasmid-borne mobile colistin resistance ( Staphylococcus aureus, followed closely by Escherichia coli, Streptococcus pneumoniae and Klebsiella pneumoniae will cause, in the worst scenario, 10 million deaths every year and a loss of 3.8 percent of the annual global gross domestic product (GDP) by 2050 . A recenAlthough bacterial AMR represents a threat to humankind, pharmaceutical research and development have failed to identify and deploy new antibiotics that can reduce and minimize this substantial danger . In thismcr, poses a significant threat because of the ability of these plasmids to move between different bacterial species through horizontal gene transfer have been detected worldwide in bacteria isolated from humans, animals, foods, the environment, and farms and the European Nucleotide Archive (ENA)] to identify specific patterns related to virulence genes, plasmid content and antibiotic resistance genes, as well as their phylogeny and their distribution with their origin and geographical location.The democratization of whole-genome sequencing (WGS) techniques and the generation of bioinformatic platforms that allow a deep characterization of pathogens can potentially transform epidemiological surveillance and disease control systems worldwide . Using tmcr-1 harboring E. coli were obtained from NCBI and ENA databases from 26 countries from four continents . The description of the sequences are included in One hundred and twenty-three WGS sequences of E. coli virulence genes was carried out using VirulenceFinder v2.0 were determined using MLST v2.0 . The preder v2.0 . The ideder v2.0 . The corder v2.0 . The obtder v2.0 .post hoc for pairwise comparisons between groups, and p-values <\u20090.05 were considered significant. The data analysis for this paper was generated using the Real Statistics Resource Pack software (Release 7.6). Copyright (2013\u20132021) Charles Zaiontz.The distribution of the genotypes between hosts and location was calculated using a one-way analysis of variance (ANOVA). Tukey\u2019s multiple-comparison test was performed This work does not involve the use of human subjects or animal experiments.mcr-1 harboring E. coli isolates evaluated in this study is shown in E. coli sequences are shown in mcr-1 harboring E. coli were not entirely correlated with the origin of the samples and the geographical precedence.The phylogenetic tree of the core-genome multilocus sequence type allelic profiles of p\u2009<\u20090.05, one-way ANOVA with Tukey\u2019s multiple-comparison test). Globally, one of the most frequent virulence genes detected was the virulence factor terC (resistance to tellurium), which had an incidence of 96%. However, even though the Ter operon role was associated with pathogenicity and stress response, the biochemical mechanisms of this association are still unclear (gad factor (glutamate decarboxylase system) had an incidence of 77%, and traT (serum resistance in E. coli) had an incidence of 76% of the evaluated genomes. The presence of gad genes was suggested as a prescreening marker for the detection of pathogenic E. coli groups and this was applied to evaluate the load of pathogens in environmental samples was generally homogeneous ( unclear . The res samples . Both georldwide .sitA virulence vector (prevents unproductive conjugation) was recorded in all of the animal samples from 2014 and 2018; sitA was applied as an excellent genetic marker for the identification of avian pathogenic E. coli, considering their significantly difference in the expression on pathogenic vs. non-pathogenic strains (E. coli (ExPEC) isolates worldwide was detected with considerable frequency in comparison with isolates from human and animal origin. lpfA was commonly associated with enteropathogenic E. coli (lpfA gene (p\u2009<\u20090.05) in E. coli from agricultural soils in comparison with the isolates from non-agriculture origin . The presence of this gene was significatively higher in extraintestinal pathogenic E. coli, and their presence might be a disadvantage for colonization in humans and animals was highly correlated with isolates from human, animal, and food/feed origin. This gene could influence capsule production in bacteria .Another virulence gene frequently detected in environmental samples was animals . In contbacteria . The resmcr-1-harboring E. coli commonly show different antibiotic resistance patterns, and the detection of multiresistant genotypes in these isolates increases their epidemiological relevance significantly. Among the analyzed genomes, the variant mcr-1.1 is the most frequently detected globally. In western Asia, the variant mcr-1.26 from different STs was detected in avian isolates from Lebanon. The same variant was detected in isolates from highly polluted water samples from Brazil that included highly-pathogenic STs (mcr-1.5 was detected in Bolivian E. coli isolate of human origin (NCBI Accession SAMN08290415). Nowadays, no additional information about this isolate has been published.The enic STs . The varmcr-1.1 and mcr-9. Additionally, this isolate encodes the extended-spectrum \u03b2-lactamases (ESBL) genes blaSHV and blaCMY. However, no additional information about this isolate was published until now. The co-occurrence of mcr gene variants is not frequently detected; however, there are a few recent reports from an E. coli isolate of human origin in Thailand shows the co-harboring of two mobile colistin resistance genes: mcr-harboring bacteria possess multidrug resistance phenotypes and their subsequent antibiotic-resistance genes (ARGs). No significant differences were identified according to the sample origin . However, significant differences were identified between geographic locations in the detection of ARGs . This was specifically found between the American and African groups and South and East Asian and African groups. Among the vast plethora of ARGs detected in silico, the most frequent genes detected were the ESBL genes. blaTEM was seen in higher proportions in environmental (93%), human (68%), food/feed (71%), and animal (53%) samples. According to the geographical distribution, this gene was detected worldwide in 50\u2013100% of samples, showing a higher presence in Asian and African isolates. Another ESBL gene with a considerable presence was blaCTX-M which has shown extended dissemination in the worldwide community since the 2000s and with lower frequency in human and animal samples. The same trend was observed in blaCTX-M-65 and blaCTX-M-15, showing higher positive results in environmental samples compared to animal and human isolates. The variants CTX-M-65 and CTX-M-55 showed a familiar presence in isolates from Asia, Europe, and America. Several questions still arise to elucidate these trends that link the environmental sources as a potential reservoir or origin of these CTX-M subtypes and the acceleration of the dissemination in the community.Most he 2000s . Among tEnterobacterales carbapenem-resistant as critical represents a severe threat to the healthcare system worldwide . The WHOntil now . The incmcr-E. coli, mainly in human and animal origin isolates, are the blaCARB and the blaCMY genes. The carbenicillinase resistance gene blaCARB, earlier known as blaPSE, is commonly associated with a chromosomal cassette in different bacterial genera, including Acinetobacter, Salmonella, Escherichia, and Pseudomonas was detected in the 28% of the evaluated isolates, showing a homogeneous distribution in animal, human and environmental samples. However, a higher presence of this gene was observed in the isolates of southeast and western Asia compared to America and Europe. In addition, the presence of acquired ARGs to the trimethoprim gene dfrA (dihydrofolate reductase) variants was considerably higher in Asian countries compared with European and American countries. In Enterobacterales, these genes are usually part of mobile gene cassettes associated with integrons that was considered the principal vector for horizontal gene transfer (HGT) of the variants of to date . Unfortumcr-1 variants and other ARGs , a global partnership coordinated by TDR, the UNICEF, UNDP, World Bank, WHO Special Program for Research and Training in Tropical Diseases hosted at the World Health Organization. The specific SORT IT program that led to this research article included an implementation partnership of TDR and the Pan American Health Organization (PAHO), the WHO Country offices of Colombia and Ecuador; the Ministry of Health and Social Protection, Colombia; Food and Agriculture Organization, Sierra Leone; Sustainable Health Systems, Freetown, Sierra Leone; The Tuberculosis Research and Prevention Center Non-Governmental Organization, Armenia; The International Union Against Tuberculosis and Lung Diseases, Paris, France and South East Asia offices, India; Institute of Tropical Medicine, Antwerp, Belgium; Damien Foundation, Belgium; Indian Council of Medical Research-National Institute of Epidemiology; Jawaharlal Institute of Postgraduate Medical Education and Research (JIPMER); GMERS Medical College Gotri Vadodara Gujarat, India; India Medical College Baroda, Gujarat, India; Sri Manakula Vinayagar Medical College, India; Public Health, Ontario; The Universidade Federal de Ciencias de Saude de Porto Alegre, Brazil; Universidade de Brasilia, Brazil; Universidad de Concepcion, Chile, Universidad de los Andes, Colombia; Universidad Pontificia Bolivariana, Colombia; Universidad Pedag\u00f3gica y Tecnol\u00f3gica de Colombia; The Central University of Ecuador and; California State University of Fullerton, United States; The Autonomous University of Yucat\u00e1n, M\u00e9xico.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/WC-C designed the investigation. KR, AM, JM, and CB-C performed the preliminary analyses of the data. WC-C, KR, AM, and JM performed genomic analysis, including genome assemblies, MLST, antibiotic resistance genes, plasmid and cgMLST analysis. MR, NO-G, TS, CD, and AH collaborated in the protocol designing according to Structured Operational Research and Training Initiative SORT IT . WC-C, AH, NO-G, TS, CD, CB-C, and MR obtained the approbation from the Ethics Advisory Groups from PAHO and The Union. WC-C supervised the entire work. WC-C, AM, JM, and AH wrote the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Growing studies have shown that insulin resistance (IR) is associated with cardiovascular disease (CVD), while the association between IR and subclinical myocardial injury (SC-MI) remains unclear. Hence we aimed to assess the association between IR and SC-MI.In this cross-sectional study, we enrolled 6043 individuals free from CVD from the third National Health and Nutrition Examination Survey. A novel metabolic score for insulin resistance (METS-IR) was used as alternative markers of IR. Multivariate logistic regression and restricted cubic spline were performed to evaluate the associations between METS-IR and SC-MI.The multivariate logistic regression analysis showed that after adjusting for cardiovascular metabolic risk factors, higher METS-IR was independently correlated with higher risk of SC-MI . Restricted cubic spline indicated that there was a J-curve connection between METS-IR and SC-MI. Threshold effect analysis ascertained an inflection point of 37 of METS-IR. The ORs (95% CIs) of per 10-unit increase of METS-IR for SC-MI were 0.707 and 1.327 on the left and right sides of\u00a0the inflection point (P < 0.05), respectively. Subgroup analysis showed that the association between METS-IR and SC-MI was only statistically significant in participants without diabetes.METS-IR was nonlinearly related to SC-MI in the general population without CVD. As an early asymptomatic myocardial injury, subclinical myocardial injury (SC-MI) is often hidden and easily overlooked in clinical work. It is reported that SC-MI can be diagnosed by a non-invasive, convenient and repeatable electrocardiograph (ECG) score, namely cardiac infarction/injury score (CIIS) . As a scIt is reported that insulin resistance (IR) is involved in the occurrence of CVD . At presConsequently, we aimed at investigating the association between METS-IR and SC-MI in this cross-sectional study.All participants in this study came from a national survey aimed at investigating the nutritional and health status of children and adults in the United States, namely the third National Health and Nutrition Examination Survey (NHANES III). After excluding individuals with CVD, major ECG abnormalities and absence of TG, FPG, HDL-C, BMI and CIIS data, 6043 individuals were finally enrolled in our study Figure\u00a01The staff of NHANES III collected demographic information from all participants through standardized questionnaires. Demographic variables enrolled in this study include age, sex, race, smoking history, prevalence of hypertension and diabetes. In this study, we divided races into four groups: non-Hispanic White, non-Hispanic Black, Mexican American and others. Those who claimed to have smoked more than 100 cigarettes were classified as smokers. The history of hypertension and diabetes were determined based on the self-reported situation of participants during the interview. The above-mentioned staff registered the BMI and blood pressure of each individual through standardized physical examination procedures. BMI was calculated according to the accepted formula, that is, weight (kg) divided by the square of height (m). Professionals of NHANES III measured the laboratory parameters of all individuals by standard biochemical analysis methods. The indicators used for this study included FPG, hemoglobin A1c (HbA1c), TG, TC, HDL-C, low-density lipoprotein cholesterol (LDL-C), uric acid (UA), blood urea nitrogen (BUN), fibrinogen, creatinine and C-reactive protein (CRP).2]/ln(HDL-C [mg/dL]), in which the blood indicators were derived from the venous blood of participants who fasted for more than 8 hours \u00d7 BMI Table\u00a05.Our study was the first report on the association between METS-IR and SC-MI. The findings showed that there was an independent J-type correlation between METS-IR and SC-MI. On the left side of inflection point, the risk of SC-MI decreased with the increase of METS-IR, while on the right side of the inflection point, the risk of SC-MI increased with the increase of METS-IR.It is well known that IR, as the core component of metabolic syndrome, is also a risk factor for metabolism-related diseases. At present, a large number of studies have focused on the effects of IR on metabolism-related diseases. Nevertheless, there are many kinds of markers of IR, so it is difficult to determine which marker has the best performance in predicting these diseases. EHC is considered to be the gold standard for the diagnosis of IR, while it has been gradually abandoned because of its complex and expensive shortcomings . SecondlThough we proved the association between METS-IR and SC-MI, the mechanism remained unknown. Based on the published literature, we found that there might be several potential mechanisms that mediated their association. For instance, there was evidence that IR promoted visceral obesity, dyslipidemia, endothelial dysfunction and elevated inflammatory markers, which were also risk factors for myocardial injury . FurtherAlthough this study acquired unexpected findings, there were still several uncontrollable limitations. For example, we failed to identify the causal association between independent and dependent variable in this cross-sectional study. In addition, there were presently a variety of markers of IR, including insulin-derived and non-insulin-derived indices, while our study mainly explored the association between METS-IR and SC-MI and failed to compare which marker was more superior in the diagnosis of SC-MI in this population. Additionally, there might be other confounding factors, such as diet and drugs. Finally, only adults from the United States were enrolled in this study, consequently, the findings might not apply to other countries and populations.In summary, this study confirmed that METS-IR, a novel non-insulin-based metabolic score for IR, was nonlinearly related to SC-MI, which further highlighted the role of IR in the occurrence and development of CVD.https://wwwn.cdc.gov/nchs/nhanes/Default.aspx.Publicly available datasets were analyzed in this study. This data can be found here: The studies involving human participants were reviewed and approved by National Center for Health Statistics of the Center for Disease Control and Prevention Institutional Review Board. The patients/participants provided their written informed consent to participate in this study.ZW and WL conceived and designed the study. WL and JL were responsible for the management and retrieval of data, contributed to initial data analysis and interpretation. ZW drafted the initial manuscript. NL revised the manuscript. NL was the guarantor of this work and had full access to all the data in the study and take responsibility for its integrity and the accuracy of the data analysis. All authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Correction to: Archives of Public Health 78, 75 (2020)10.1186/s13690-020-00457-4Following publication of the original article , the autThe original version was:While each cohort center received the ethical approval from local universities, for the purpose of this study and pooling all PERSIAN data, the ethics committee of Kermanshah University of Medical Sciences approved the study (IR.KUMS.REC.1397.187).The correct version is:While each cohort center received the ethical approval from local universities, for the purpose of this study and pooling all PERSIAN data, the ethics committee of Kermanshah University of Medical Sciences approved the study (IR.KUMS.REC.1397.866).The original article has been"} +{"text": "British Journal of Cancer 10.1038/s41416-022-02050-8, published online 17 November 2022Correction to: The original version of this article contained an error in the funding section. This work was supported by RSF-2017-00000557 funding from Veneto Region . Role of the funder/sponsor: The funder had no role in the design and conduct of the study; collection, management, analysis, and interpretation of the data; preparation, review, or approval of the manuscript; and decision to submit the manuscript for publication. The original article has been corrected."} +{"text": "Some of the most direct and brutal effects of the COVID-19 pandemic are experienced by health care professionals who are working in demanding environments while having to deal with their own fears of infection and mortality. To assess the impact of COVID-19 on the practice and well-being of global mental health professionals, we designed a three-part, longitudinal, internet-based study. Here we present data from part 1, implemented in June-July 2020 in six languages to members of WHO\u2019s Global Clinical Practice Network composed of 15,500 mental health practitioners. The study assessed COVID-19\u2019s impact on: work circumstances; occupational well-being; use and transition to telehealth; and expectations, needs and recommendations. 2,505 mental health professionals from 126 countries responded to the study (47% psychiatrists). 93.7% of respondents were currently practicing and 70.9% continued to see patients in person. The impact on clinical workload varied in terms of direction and extent depending on type of service provided and country of practice. Most participants had started or increased their use of telehealth services, and we identified a need for training to support telehealth use. Overall, clinicians scored high on well-being indices. However, a subset scored above the cutoff for low well-being and reported a significant number of post-traumatic symptoms. Five factors affected work-related stress: fear of infection, severe COVID-related events, life disruption, lack of adequate protection and role disruption. Data from this study will provide information relevant for the design, development, and integration of mental health services in the continuing pandemic, and in similar future scenarios.No significant relationships."} +{"text": "Scientific Reports 10.1038/s41598-022-21965-z, published online 07 November 2022Correction to: In the original version of this Article, the Acknowledgements section has been removed, as the funder has no role in the data collection, analysis, and interpretation of the data.The original Article has been corrected."} +{"text": "Early detection is crucial to control the progression of Alzheimer's disease and to postpone intellectual decline. Most current detection techniques are costly, inaccessible, or invasive. Furthermore, they require laborious analysis, what delays the start of medical treatment. To overcome this, researchers have recently investigated AD detection based on electroencephalography, a non-invasive neurophysiology technique, and machine learning algorithms. However, these approaches typically rely on manual procedures such as visual inspection, that requires additional personnel for the analysis, or on cumbersome EEG acquisition systems. In this paper, we performed a preliminary evaluation of a fully-automated approach for AD detection based on a commercial EEG acquisition system and an automated classification pipeline. For this purpose, we recorded the resting state brain activity of 26 participants from three groups: mild AD, mild cognitive impairment (MCI-non-AD), and healthy controls. First, we applied automated data-driven algorithms to reject EEG artifacts. Then, we obtained spectral, complexity, and entropy features from the preprocessed EEG segments. Finally, we assessed two binary classification problems: mild AD vs. controls, and MCI-non-AD vs. controls, through leave-one-subject-out cross-validation. The preliminary results that we obtained are comparable to the best reported in literature, what suggests that AD detection could be automatically detected through automated processing and commercial EEG systems. This is promising, since it may potentially contribute to reducing costs related to AD screening, and to shortening detection times, what may help to advance medical treatment. Alzheimer's disease (AD) is a neurogenerative disease that represents 60\u201370% of the dementia cases for AD detection. EEG is a neurophysiology technique to measure the electrical activity of the brain through electrodes placed on the scalp. EEG is portable, non-invasive, and affordable compared to medical imaging procedures. Consequently, it represents a promising approach for the detection of neurological diseases. Indeed, researchers have recently combined EEG signal processing and machine learning algorithms to discriminate AD and MCI patients from age matched controls , lack of cooperation (participants 9 and 18), and presence of a condition that may have impacted the analysis (participant 12), we excluded the data from five participants. Consequently, we only studied the recordings from 21 participants. The head of the CBNU at Hospital Universitario Virgen de las Nieves de Granada recruited the participants the week before the experiment. First, we asked the participants to read and sign the informed consent. Then, we briefed them on the study details and the basics of EEG acquisition. It is worth to note that the participants of this study were recruited with two objectives in mind: (1) go through a cognitive test, and (2) participate in the study reported in this paper. The cognitive test consisted of two tasks of approximately 10 min whose details are out of the scope of this paper. In parallel, we recorded 3 min of the eye-open resting state activity of the participants in three key moments: before the first task, after the first task, and after the second task. To avoid edge effects, we only considered the central 2-min window of each recording. For the analysis reported in this paper, we concatenated the three recordings, thus, we ended up with an effective 6-min EEG recording per participant.For the acquisition, we used the Versatile system by Bitbrain , a commercial wearable device consisting of an EEG cap with semi-dry electrodes and a Bluetooth acquisition module that works at a fixed sampling rate of 256 Hz. This sampling rate has been previously used in related studies in the five main EEG bands, Hjorth complexity (HC), and spectral entropy (SE). Consequently, we extracted a total of 112 features per epoch. We considered these features since they have been already applied in analogous studies , theta (4\u20138 Hz), alpha (8\u201313 Hz), beta (13\u201330 Hz), and gamma (>30 Hz). In Equation (2), \u03c3s, f represents frequency, and S represents the normalized power spectrum.In Equation (1), To create the feature matrix, we vertically concatenated the features extracted for each participant. Consequently, we ended up with a feature matrix with 112 columns (features) and as many rows as the total number of epochs from all the participants see . Then, wTo solve the two binary classification problems examined in this study, we used scikit-learn Python module are estimated using the training set, and subsequently, those parameters are applied on the test set. This avoids overfitting and prevents from artificial positive bias.In this section, we present the results that we obtained for the two binary discrimination problems evaluated in this study: mild AD vs. NC and MCI-non-AD vs. NC, based on the 16-channel resting state EEG recordings that we acquired (see Section 2.2).The goal of this study was to perform a preliminary evaluation of an automated approach for the discrimination of AD cohorts. For this purpose, we recorded the EEG of a group of volunteers, and we proposed an approach based on a commercial EEG device with a reduced montage and an automated classification pipeline. The preliminary results that we obtained from a reduced sample suggest that such an approach can precisely discriminate mild AD and MCI-non-AD participants from healthy controls.In In terms of recording conditions, most studies in literature typically use between 17 and 32 electrodes .via leave-one-subject-out cross-validation. The results that we obtained are comparable to the best reported in literature, what is promising toward the implementation of automated AD detection approaches based on commercial acquisition devices. Since we analyzed a reduced sample size, further studies must evaluate the presented approach on larger samples to validate the conclusions yielded in this paper. Nonetheless, the results obtained in this work are promising, as they suggest that AD detection could be performed automatically using a few minutes of resting state EEG activity and a commercial acquisition device. With this in mind, we believe the future inclusion of this kind of approaches into AD screening practice could contribute to reducing the costs linked to AD detection, and also enable the early detection of the disease, what could potentially advance medical treatments. Nonetheless, further research is also required to investigate the feasibility of automated EEG-based approaches in participants that suffer from additional pathologies whose symptoms overlap with those of dementia. This is often the case in daily-life neurological practice, hence, such cases must be comprehensively studied before future patients can take advantage from fast, accurate, and affordable detection techniques that enhance their standards of living.In this paper, we preliminarily evaluated the feasibility of an automated pipeline based on EEG activity for the discrimination of mild AD and MCI-non-AD. For this purpose, we recorded the resting state activity of a reduced sample of volunteers using a commercial EEG device, and we designed our approach to automatically perform signal processing, feature extraction, and classification. We applied the Autoreject algorithm and ICA for artifact rejection. Then, we estimated the relative power, the Hjorth complexity, and the spectral entropy of the preprocessed epochs. Lastly, we assessed two binary classifiers (SVM and LR) for the discrimination of the three cohorts of interest The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Hospital Universitario Virgen de las Nieves de Granada. The patients/participants provided their written informed consent to participate in this study.EP-V and ML-G: conceptualization and methodology. EP-V: software, formal analysis, investigation, resources, data curation, and writing\u2014original draft preparation. EP-V, ML-G, and CM: validation. EP-V, ML-G, CM, RV-C, IC-M, and SL-A: visualization and writing\u2014review and editing. ML-G and CM: supervision. ML-G, CM, RV-C, IC-M, and SL-A: project administration. All authors have read and agreed to the published version of the manuscript.This work was supported by project PID2021-128529OA-I00 Proyectos de Generaci\u00f3n de Conocimiento 2021 funded by the Spanish Ministry of Science, Innovation and Universities, by European Regional Development Funds, and by project B-TIC-352-UGR20, and also co-funded by the Operative Program FEDER 2014\u20132020 and the Economy, Universities and Science Office of the Andalusian Regional Government.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Hantavirus is known to be transmitted from rodents to humans. However, some reports from Argentina and Chile have claimed that the hantavirus strain Andes virus (ANDV) can cause human-to-human transmission of the disease. The aim of this systematic review was to assess the evidence for human-to-human transmission of hantavirus.We searched PubMed (inception to 28 February 2021), Cochrane Central, Embase, LILACS and SciELO (inception to 3 July 2020), and other sources. We included studies that assessed whether interpersonal contact with a person with laboratory-confirmed hantavirus infection led to human-to-human transmission. Two reviewers conducted screening, selection, data extraction, and risk of bias assessment.Twenty-two studies met the inclusion criteria. Meta-analysis was not possible due to heterogeneity. With the exception of 1 prospective cohort study of ANDV in Chile with serious risk of bias, evidence from comparative studies (strongest level of evidence available) does not support human-to-human transmission of hantavirus infection. Noncomparative studies with a critical risk of bias suggest that human-to-human transmission of ANDV may be possible.The balance of the evidence does not support the claim of human-to-human transmission of ANDV. Well-designed cohort and case-control studies that control for co-exposure to rodents are needed to inform public health recommendations. The balance of the evidence does not support the claim that human-to-human transmission of hantavirus infection occurs. Well-designed cohort and case-control studies that control for co-exposure to rodents are needed to inform public health recommendations. The 1993 hantavirus outbreak in the southwest of the United States [Orthohantavirus) cause 2 zoonotic diseases in humans, which are clinically manifested in 2 distinct forms: (1) hemorrhagic fever with renal syndrome (HFRS), which occurs in Europe and Asia; and (2) HPS in the Americas. Typically, HFRS causes a mild-to-moderate infection presenting fever, headaches, and gastrointestinal symptoms, with a progression to hypotension and acute renal failure, but with a low fatality rate (1%\u201315%) [Hantaviruses . HPS, by(1%\u201315%) , 5.% . HPS, Both diseases are carried by rodents and each particular species of hantavirus has a particular rodent as its intermediate host. Environmental conditions that favor the reproduction and spread of rodents in endemic areas are known to increase the incidence of the disease. Humans typically acquire the disease through the inhalation of aerosolized excreta or secreta from infected rodents.Human-to-human transmission of the Andes virus (ANDV) was first claimed to have occurred as part of the 1996 outbreak in southern Argentina , and sinIt is important to determine whether there is in fact sufficient evidence for human-to-human transmission of the disease. If so, this may require recommendations regarding infection prevention and control measures in health facilities and in household settings. Thus, the objective of this systematic review was to assess the evidence for human-to-human transmission of hantavirus.High-quality systematic review methods were used and the We searched Cochrane Central, Embase, Latin American and Caribbean Health Sciences Literature (LILACS), PubMed, and Scientific Electronic Library Online (SciELO) from inception to date of search (3 July 2020). The PubMed search was updated 28 February 2021. In addition, Google was searched using the same key words for gray literature. Reference lists of included studies, key literature reviews, and Pan American Health Organization/World Health Organization documents, such as country-level guidelines, were scanned for relevant studies. Google Scholar was used to search for articles that cite key articles. Contact was made with known hantavirus experts to identify both published and unpublished studies, though the response was poor, possibly due to the fact that many of the key studies were published >20 years ago.The search terms included medical subject heading (MeSH) terms (where relevant for the database) and text words. Searches were conducted by 1 review author (M. M. H.) and references were imported into Endnote. The search strategy and results for each of the databases are included in either reviewer was retrieved and assessed against the inclusion criteria by 2 reviewers (M. M. H. and L. S. L.) independently. Disagreements regarding eligibility of studies were resolved via discussion and consensus, with consultation with a third reviewer if needed (J. T./L. R.).The screening of the titles and abstracts against the inclusion criteria was condTwo reviewers independently extracted all relevant data from the included papers (M. M. H. and L. S. L.) into a Microsoft Excel spreadsheet. Differences were resolved by discussion and consensus, with consultation with a third reviewer (J. T./L. R.) if needed.Data extracted included study ID, year of study, country, region/city, setting of study, hantavirus species, mouse species, study type, type of outbreak, participants , exposure , outcomes, results, and other relevant information .The risk of bias (RoB) of included studies was assessed by 1 reviewer (M. M. H.) and verified by a second (L. S. L.), with consultation with a third reviewer in the case of disagreements (J. T./L. R.). The Risk of Bias in Non-randomized Studies\u2014of Exposures (ROBINS-E) tool was used for nonrandomized comparative studies , 20. TheData were synthesized in both tabular and narrative formats. A meta-analysis was not possible due to heterogeneity in the studies and outcomes measured.We identified 1321 records after removal of duplicates . The iniNo randomized controlled trials were found. The studies included a total of 5190 participants: 4179 in the comparative designs and 1011Of the 8 comparative study designs, 6 were classified as having a critical RoB , 36, 38 P = .66) [2 = 0.03, P = .86) [Three of the 8 comparative designs were conducted in Argentina or ChileP = .66) . The secP = .86) . While tP = .86) . The 5 cP = .86) , ParaguaP = .86) , China [P = .86) , and theP = .86) showed nOf the 14 noncomparative studies, 7 were conducted in Argentina , 26, 32 Similar to the case in Argentina, the 2 studies investigating clusters in Chile did find some evidence of human-to-human transmission, with up to 5 likely cases of human-to-human transmission reported , 35. HowThis systematic review presents a comprehensive and systematic evaluation of the research on human-to-human transmission of hantavirus. Despite claims of authors from Argentina and Chile of the existence of human-to-human transmission of ANDV, the balance of the evidence does not support this claim. Evidence from comparative studies (the strongest level of evidence available) does not support human-to-human transmission of hantavirus infection , 36, 38,The absolute number of cases that may be attributable to human-to-human transmission is low and needs to be contrasted with the much larger number of cases that did not lead to human-to-human transmission, even with similarly close contact in community or health facility settings. For example, the cohort study by Ferres and colleagues included 476 household contacts, of which only 16 subsequently developed HPS, 3 of which were claimed to be definitely due to human-to-human transmission and 9 probably due to human-to-human transmission; the remaining 460 household contacts did not . Their mB and Supplementary Figure 3 in Mart\u00ednez et al [Analysis of the evidence from noncomparative studies suggests a possibility of human-to-human transmission of ANDV in some parts of Argentina and Chile. However, as noted above, these studies are not able to make causal inferences and are limited to a few outbreaks, and the findings of some outbreaks have been published across different reports, with some inconsistencies in the accounts . Yet, tWhile the use of genetic sequencing data to confirm human-to-human transmission has been used in investigations of clusters in Argentina and Chile , 32, 35,A key strength of this review was the use of high-quality systematic review methods that incThe RoB in included studies is a particular concern when trying to determine whether human-to-human transmission of hantavirus is the best explanation for infection in other human contacts. A randomized controlled trial, where subjects are randomized to exposure to another human with hantavirus or to a nonexposure control group, in theory, would constitute the ideal study design. However, ethical and feasibility concerns make this kind of study impractical at this stage of our understanding of the problem. The next best study designs fall in the observational design category but also require the use of a control (or comparison) group, such as in a cohort or case-control design\u2014which are commonly used designs for outbreak investigations . These wWhat are the public health implications of these findings? Given the high case fatality rate for HPS, the precautionary principle could be invoked to recommend action to implement infection prevention control containment measures in cases of suspected hantavirus infection in health facilities and household settings where ANDV is present. This could include the implementation of standard precautions and rational and optimized use of personal protective equipment, including the use of filtering facepiece respirators, or respirators by health workers to prevent possible infection by inhalation of droplets or aerosolized virions. However, given the limited number of cases and the high RoB in included studies, it may be prudent to accompany such a recommendation with a strategy to gather better research evidence to answer the question.Future studies should include a control group, such as in a cohort or case-control design, and conduct a multivariate analysis to control for potential confounders, especially factors related to the potential risk of environmental exposure to the excreta/secreta of infected rodents. Care should be taken to measure the duration, type, and timing of contact with the infected person and with infected rodents. Outcome measures should include laboratory measures such as virology, serology, or immunohistochemistry. Molecular epidemiology analysis, including genetic sequencing of rodent and human specimens, may also be useful when cases have occurred outside the affected geographic area.In conclusion, this systematic review has shown that the evidence for human-to-human transmission of hantavirus is weak, specific to ANDV, and limited to some parts of Argentina and Chile. Due to the high case fatality rate for HPS, it may be prudent to recommend infection prevention and control measures in cases of suspected hantavirus infection\u2014but this needs to be accompanied by the gathering of better research evidence.The Journal of Infectious Diseases online. Supplementary materials consist of data provided by the author that are published to benefit the reader. The posted materials are not copyedited. The contents of all supplementary data are the sole responsibility of the authors. Questions or messages regarding errors should be addressed to the author.Supplementary materials are available at jiab461_suppl_Supplementary_MaterialsClick here for additional data file."} +{"text": "The proportion of the world's elderly population continues to rise, and the treatment and improvement of neurodegenerative diseases have become issue of public health importance as people live longer and many countries have aging populations. This systematic review aims to discuss the effects of dance movement therapy (DMT) on motor function, cognitive deficit, mood, and quality of life in people with neurodegenerative diseases, such as Parkinson's disease (PD), mild cognitive impairment (MCI), Alzheimer's disease (AD).Two reviewers independently conducted systematic search on the Cochrane library, PubMed database, Web of Science Core Collection database, and Physiotherapy Evidence database until February 1, 2022. Only systematic analyses and randomized controlled trials were included and further analyzed.Thirty-three studies on PD, 16 studies on MCI, 4 studies on AD were obtained. This systematic review found that DMT substantially improved the global cognitive function, memory, and executive function on the population with MCI. Compared with the non-dance group, DMT remarkably improved general disease condition, balance, and gait for individuals with PD. The evidence of the efficacy of DMT on AD is insufficient, and further research is needed.DMT can effectively improve the motor function and cognitive deficits in neurodegenerative diseases. Positive effects of DMT on the mood and quality of life in ND patients are controversial and require further evidence. Future research on the effects of DMT on AD requires scientific design, large sample size, long-term comprehensive intervention, and clear reporting standards.www.osf.io/wktez, identifier: 10.17605/OSF.IO/UYBKT. Eurostat forecasts show that the proportion of the population over 60 years old will reach 35%, and the number of people aged 65 and over with Alzheimer's disease (AD) in the United States may increase to 13.8 million by the middle of this century , 2020. ADance is a rhythmic movement that has been choreographed or improvised in advance and usually performed with music. The American Dance Therapy Association (ADTA) defines dance movement therapy (DMT) as \u201cthe psychotherapeutic use of movement as a process that promotes the emotional, social, cognitive, and physical integration of the individual.\u201d Formal DMT can be defined as active interventions or receptive interventions (such as watching stage plays) by qualified dance therapists (groups). The meta-analysis results of Fong Yan et al. confirmeAt present, there are various types of dance-movement therapy and widely accepted by groups of all ages. For example, the Videogame-Based Dance Exercise Program attracts many young people, and Chinese-style square dance is also recognized by middle-aged and elderly groups. In the foreseeable future, there will be a variety of ways for people to access dance movement therapy, no longer limited to one-on-one or one-to-many treatment modes between dance therapists and patients. This also provides one of the treatment options for at-home rehabilitation for the elderly population who have limited cognition, activity and social skills due to aging. However, extant studies only summarized some aspects of the effectiveness of DMT for a certain NDs. A comprehensive overview and interpretation of the benefits and potential mechanisms of DMT in the rehabilitation of major neurodegenerative diseases with aging as a major risk factor are needed. We conducted an integrated review of systematic analyses and RCTs of DMT in patients with PD, MCI, AD and discussed the underlying mechanisms of the efficacy of DMT.10.17605/OSF.IO/UYBKT.This protocol of this systematic review has been registered in the Open Science Framework (OSF) systematic review database, international prospective register for systematic review under the registration via the PubMed database, Web of Science Collection database, Cochrane library, and Physiotherapy Evidence Database (PEDro). No limit was set on data search. The key words of search strategies were as follows: \u201cdance,\u201d \u201cAlzheimer's disease,\u201d \u201ccognitive impairment,\u201d \u201cParkinson's disease,\u201d . This process was performed because of differences in disease type, intervention group, and primary outcome assessment in the included studies. The effects of DMT were compared with other exercises, usual care, and treatment involved in the meta-analyses and RCTs in accordance with the clinical features of each disease.A total of 262 papers overlapped out of the 721 papers extracted in the preliminary search. A total of 295 reports were excluded because of the following reasons: (1) the experimental design was not a systematic analysis or RCT; (2) the type of participants included in the study was complex ; (3) no control group; (4) no comparison or missing results were provided; (5) study was not completed. A total of 130 articles were retained for further research, and 73 articles were excluded for reasons similar to those described above. Thirty-three RCTs and 24 systematic analyses were included. As shown in MCI was introduced as the intermediate stage between healthy aging with mild cognitive changes and dementia and MoCA (p = 0.03) scores compared with usual care. Chang et al. , Montreal Cognitive Assessment (MoCA), and Function and Cognitive Assessment Test (FUCAS) were used to assess the global cognitive level of MCI patients. Three high-quality systemic and meta-analysis studies . The results of Doi et al. (p = 0.009) and increase the right and total hippocampal volumes in the individual with MCI. Remarkable cortical thickening was observed in the right inferior temporal, fusiform, and lateral occipital regions of MCI participants undergoing a 6-month dance intervention at autopsy , and standard or other physical therapy . The meta-analysis results showed that Tango is more advantageous in improving balance and gait speed in individuals with PD by comparing the effects of Tai chi, Qigong, yoga, and resistance training on PD in terms of gait velocity, TUG, and BBS and increased pelvic rotation (p = 0.036) in PD patients when turning around . Cognitive dual-task evaluation by dual-task TUG showed that PD patients in the dance groups had better improvement , and the Mental Rotation Task (response time: p < 0.001).Four systematic reviews and meta-analyses explored the efficacy of DMT on the overall cognitive level of PD patients, where three of them confirmed that the DMT group is better than the control group . In a 12-week Sardinian dance RCT, the significant time*group interactions for depression (p < 0.001) and apathy (p = 0.016) were observed in the dance group . Compared with usual care, DMT provided PD patients with a large number of new social activities (p = 0.003) in PD patients remains controversial. Compared with the non-dance group, more than half of the meta-analyses compared with the conventional treatment group, thereby improving the quality of life of AD patients and reduced the burden on nursing staff . These studies had small sample sizes and insufficient follow-up with hidden allocation and blinding. Current studies do not provide sufficient evidence to explain the whole therapeutic mechanisms of DMT. Additional high-quality, large-scale intervention studies and multimodal studies combining behavioral outcome measures with neuroimaging and neuroendocrine markers are needed to elucidate the effects and mechanisms of dance exercise therapy on NDs.The proportion of the elderly population in the world continues to increase, and the loss of independence of the elderly due to NDs leads to an increased social burden. Thirty-three RCTs and 24 systematic analyses reported the effects of DMT in NDs. DMT had remarkable effects on the condition, balance, and gait in PD patients worldwide. DMT significantly improved the cognitive function, memory, and executive function for patients with MCI. However, data are insufficient to fully demonstrate that DMT has a positive effect in patients with AD. Future research on the effects of DMT on AD requires scientific design, large sample size, long-term comprehensive intervention, and clear reporting standards.The original contributions presented in the study are included in the article/X-QW had substantial contributions to the conception of the study. X-QW and J-JZ designed this systematic review. C-CW and H-YX conduct the research. C-CW wrote the original draft of the manuscript. X-QW, J-JZ, H-YX, and C-CW participated in the revision of the draft. All authors read and approved the final submitted version.This study was supported by the National Natural Science Foundation of China (81871844), Shuguang Program supported by Shanghai Education Development Foundation and Shanghai Municipal Education Commission (18SG48), the Science and Technology Commission of Shanghai Municipality (grant numbers 19080503100 and 21S31902400), the Fok Ying-Tong Education Foundation of China (grant number 161092), the Talent Development Fund of Shanghai Municipal (grant number 2021081), the Shanghai Clinical Research Center for Rehabilitation Medicine (grant number 21MC1930200), and the Shanghai Key Lab of Human Performance (Shanghai University of Sport) (grant number 11DZ2261100), as well as by the Shanghai Frontiers Science Research Base of Exercise and Metabolic Health.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "In \u201cAmbient Assisted Living: Scoping Review of Artificial Intelligence Models, Domains, Technology, and Concerns\u201d :e36553) the authors noted one correction.Under \u201cAcknowledgments\u201d, the sentence:This work was part of and supported by GoodBrother, COST Action 19121\u2014Network on Privacy-Aware Audio- and Video-Based Applications for Active and Assisted Living.has been replaced by:This publication is based upon work from COST Action GoodBrother\u2014Network on Privacy-Aware Audio- and Video-Based Applications for Active and Assisted Living (CA19121), supported by COST (European Cooperation in Science and Technology).The correction will appear in the online version of the paper on the JMIR Publications website on December 20, 2022 together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "Pediatric Infectious Disease Journal, Pediatric Pulmonology, and Frontiers in Pediatrics. The top cited and co-cited journals were the New England Journal of Medicine, Nature, JAMA, Lancet Infectious Diseases, and BMJ. The network visualization maps of keywords and terms offered a global overview of the clinical and laboratory characteristics of pediatric COVID-19 patients. The bibliometric profile of the researched domain, based on analyzing a large collection of publications/data, could (i) enrich the researchers and non-researchers understanding of the field existing patterns and trends, and (ii) be useful in clinical practice (diagnostic and management) and public health policy.The literature on the COVID-19 landscape has rapidly expanded in the pandemic period. The current study undertakes a bibliometric analysis of research in the topic of the clinical and laboratory characteristics of pediatric COVID-19 cases. Our aim is to perform a comprehensive bibliometric review of current research trends and patterns of this research domain. Publications retrieved from the Web of Science Core Collection and VOSviewer were used for analysis and network visualization. We analyzed geographical distribution and temporal trends, collaboration and citation patterns of authors, institutions, and countries, and core research themes from co-occurrence of keywords and terms. The analysis showed that contributions in the research field were from 302 publications, 1104 institutions, 62 countries, and 172 journals. Many publications were authored by American and Chinese authors, and many were published in the The literature on the COVID-19 landscape has rapidly expanded in the pandemic period.Systematic reviews and meta-analysis studies are gathering existing knowledge, offering synthesized, selected, high-quality research evidence, seeking patterns and agreement or disagreement among studies results . HoweverThe bibliometric analysis method systematically studies the specific topic and determines its research growth, emerging topics discussed in different articles, and trends over time. By its indicators , the bibliometric method provides quantitative and qualitative analysis and measurement of the specific research field .Several reviews and meta-analysis summarized the clinical, laboratory, radiological features, outcome, and treatment of the SARS-CoV-2 infection at various time periods during the pandemic ,5,6,7,8.The aim of this study is to present a global overview of clinical and laboratory characteristics of pediatric COVID-19 cases using a bibliometric approach. We addressed the following research directions: to identify most frequently used keywords/terms (and their co-occurrence network) related to the research field, to identify the most productive countries and organizations, to identify sources where researchers publish their work, authorship, and collaborative patterns, and to identify influential research papers (highly cited). To the best of our knowledge, there is currently no bibliometric review related to the medical literature about the topic of clinical and laboratory characteristics of pediatric COVID-19 cases. We conducted a literature search on Web of Science Core Collection (WoS) online databases in October 2021 to identify scientific contributions, including analysis of clinical and/or laboratory characteristics of pediatric COVID-19 cases. The search identified publications that contain the terms (COVID-19 OR coronavirus OR SARS-CoV-2) and (pediatric OR children) and and in their title or abstract or keywords or keywords plus. We included only article-type documents from 2020 to October 2021. MT and IM reviewed the titles, abstracts, and full texts of articles selected by the automated search to reach a consensus on inclusion of the topic-relevant ones. We included published papers that reported original data about the clinical or laboratory features of children with COVID-19. Included study designs were case-control, cohort studies, case reports (more than one case), and case series. Some studies included pediatric patients aged up to 18 years, while others included patients aged up to 20 years. We included studies considering both situations. The process of study selection was described using a PRISMA flow diagram, as in Characteristics of each publication identified from the search include publication title, abstract, keyword, keyword plus, authorship, document type, publication year, journal title, language, journal category, and number of total citations. We used the VOSviewer science visualization software package developed by Van Eck and Waltman at the Center for Science and Technology Studies of Leiden University ,10. The Based on the literature search strategy, it was found that there are 302 publications, literature from the WoS database, comprising clinical or laboratory characteristics of pediatric COVID-19 cases.More than 1000 institutions (1104) from 62 countries/regions have reported clinical or laboratory characteristics of pediatric COVID-19 cases in their research. The most productive institutions were Huazhong Univ. of Science and Technology , the Univ. of Pennsylvania , the Children\u2019s Hospital of Philadelphia , Univ. Paris , Columbia Univ. , and Chongqing Med. Univ. . The most cited institutions were the Univ. of Pennsylvania , Huazhong Univ. of Science and Technology , Vanderbilt Univ. , Harvard Med. School , and Boston Children\u2019s Hospital . Collaboration analysis of the institutions is presented in The leading countries in the analyzed researched were the USA , China , Turkey , Italy , England , and France . The rank of the countries is detailed in Top productive and cited journals that include publication related to clinical and laboratory characteristics of pediatric COVID-19 cases are presented in The top 20 highly cited articles are presented in -Shao, Jianbo -Li, Hui -Villani, Alberto -Rossi, Paolo -Xiao, Han -Fitzgerald, Julie -Son, Mary Beth F. -Newburger, Jane -Clouser, Katharine The authors with the highest number of citations were: Keywords networks are usually used to map topics of the research field. The knowledge map of keywords co-occurrence (author keywords and keywords plus) and knowledge map of terms from publication title and abstract are presented in This study reveals the bibliometric profile of the research and visualization of connections between pieces of information in the field of clinical and laboratory characteristics of pediatric population with COVID-19. It summarizes the most frequent symptoms and laboratory parameters of the literature in the research field. The literature in the studied field is represented by more than 300 publications. On the list of top productive countries were the USA, China, Turkey, Italy, and England, while on the list of top cited countries were the USA, China, England, France, and Italy. Keyword/term-based maps show a global overview of clinical and laboratory characteristics of pediatric COVID-19 cases. A network visualization map of terms and keywords revealed the most commonly included symptoms and laboratory imaging findings among pediatric patients with COVID-19. Regarding symptoms, terms related to systemic, respiratory, gastrointestinal, neurological, olfactory, dermatological, ocular system, rheumatic, and cardiac symptoms were observed. Regarding laboratory features, terms related to inflammatory markers, immune markers, d-dimer levels, and liver enzymes were observed.Terms like multisystem inflammatory syndrome (52), MISC/MIS-C (48/17), Kawasaki disease/Kawasaki-disease/Kawasaki-like disease (11/10/4), macrophage activation syndrome/MAS (7/4), and toxic shock syndrome/TSS (8/2) are referring to studies that describe clinical or laboratory values of coronavirus disease 2019-related multisystem inflammatory syndrome in children or Kawasaki disease or TSS or MAS. Among MISC-related studies, some of them were addressing organ damage or impairment manifestations ,71, cardIn Italy, findings of an multicentric study showed aOur study had several limitations. We only included studies extracted and analyzed from the WoS database. Citations of research papers accumulate over time, being considered to be biased toward old articles (articles published recently are hardly cited (yet)). Although keyword and terms analysis offered relevant information regarding each research study, a more in-depth content analysis of the publications could provide more specific information/associations/mechanisms/etc. regarding COVID-19 pediatric features. Our study included only articles published until October 2021, though research on COVID-19 is still ongoing. In addition, we did not consider/ensure the methodological quality of the studies included in the bibliometric review.We believe this analysis provides a global scientific output of clinical and laboratory characteristics that could be useful in informed research, clinical practice (diagnostic and management), and public health policy."} +{"text": "In the future, tuberculosis (TB) will place a heavy burden on the aging population in Korea. To prepare for this crisis, it is important to analyze the disease burden trend of drug-susceptible tuberculosis (DS-TB) and multidrug-resistant tuberculosis (MDR-TB). Measuring disability-adjusted life years (DALYs) and economic burden on MDR-TB patients can help reduce the incidence of TB. Accordingly, in this study, we measured the DALYs and economic burden on DS-TB and MDR-TB patients in 2014\u20132017 using a combination of National Health Insurance claims data, Annual Report on the Notified TB data, and Statistics Korea's mortality data. The incidence-based DALY approach implemented involved the summation of years of life lost and years lived with disability. For measuring economic burden, direct and indirect costs incurred by patients were totaled. From 2014 to 2017, DALYs per 100,000 people with DS-TB were 56, 49, 46, and 40, respectively, and DALYs per 100,000 people with MDR-TB were 3, 2, 2, and 2, respectively. The economic burden for the DS-TB population from 2014 to 2017 was $143.89 million, $136.36 million, $122.85 million, and $116.62 million, respectively, while that for MDR-TB was $413.44 million, $380.25 million, $376.46 million and $408.14 million, respectively. The results showed a decreasing trend in DALYs and economic burden for DS-TB, whereas MDR-TB was still found to be burdensome without a specific trend. With respect to age, the economic burden for both DS-TB and MDR-TB was higher among men than among women till \u2264 79 years. Conversely, the economic burden for women aged \u226580 years was higher as compared to their male counterparts. In conclusion, the incidence and spread of TB in all areas of society must be suppressed through intensive management of MDR-TB in the older population. We hope that the national TB management project will proceed efficiently when the infectious disease management system is biased to one side due to the COVID-19 pandemic. Tuberculosis (TB) is a highly infectious disease , 2 that Meanwhile, drug-resistant TB (DR-TB) has emerged as an important target for TB control worldwide. Drug resistant TB can stem from improper management including poor compliance and wrong treatment as well as from direct transmission. Multidrug-resistant tuberculosis (MDR-TB) refers to tuberculosis that is resistant to at least both isoniazid and rifampicin and extensively drug-resistant tuberculosis (XDR-TB) is an extensively drug-resistant TB that is resistant to any fluoroquinolone and at least one of three injectable second-line drugs in addition to MDR-TB resistance, on drug-susceptibility testing (DST) microbiologically . ConfronSouth Korea has high TB incidence and mortality rates. The incidence of TB in South Korea is the highest among the Organization for Economic Cooperation and Development (OECD) countries . The plaBased on these statistics, TB will put a heavy burden on Korea in the future. To prepare for this crisis, it is important to analyze the disease burden trends of DS-TB and MDR-TB based on the age and sex of the affected population \u201317. The There is a lack of research for the measurement of health outcomes of MDR-TB patients with the use of a standardized methodology as a result of limited data sources and difficulties in selecting subjects. In this study, National Health Insurance (NHI)-integrated data can be used as a source of information for the calculation and comparison of the DALYs of and economic burden on TB patients with different levels of drug resistance to generate reliable evidence. Therefore, this research aimed to identify the current disease burden and trends of DS-TB and MDR-TB in South Korea and calculate the economic burden on patients utilizing NHI-integrated data from 2014 to 2017.This study was conducted using a combination of the Annual Report on the Notified TB data, NHI claims data, and Statistics Korea's mortality data. The Annual Report on the Notified TB data was prepared to analyze information pertaining to TB patients diagnosed or treated at public health centers and hospitals across the country and to plan, implement, and evaluate national TB management policies based on the results. The data were collected through the integrated disease and health management system, NHI claims data refers to a sample research data, customized research data, and health disease indicators based on evidence accumulated from the NHIS. Statistics Korea's mortality data is a compilation of death reports acquired from Eup, Myeon, Dong administrative welfare centers, and Si/Gu offices in the country (foreign missions for overseas Koreans) based on resident registration sites.In this study, MDR-TB patients and DS-TB patients were selected as participants. The description of the codes and criteria for selecting participants are as follows. Participants diagnosed with TB-related ICD-10 codes were selected . MDR-TB patients were diagnosed with U84.30 (MDR-TB), U84.31 (XDR-TB), and rifampin-only resistant TB, while other participants were diagnosed with DS-TB. The U84.30 code is assigned to patients with TB bacteria resistant to two or more anti-TB drugs, including isoniazid and rifampin. The U84.31 code is for TB patients resistant to isoniazid and rifampin, one or more quinolone drugs, and one or more of the three injections, capreomycin, kanamycin, and amikacin.The disability-adjusted life years (DALY) approach was used as a methodology in this research, and the incidence-based DALY approach was measured by totaling the values corresponding to years lived with disability (YLD) and years of life lost (YLL). One DALY indicates that 1 year of full health was lost to disease, disability, and early death; as DALYs increase, the gap with the ideal health level widens, which suggests an increase in disease burden. The incidence-based DALY approach was measured by totaling the values corresponding to YLD and YLL, and detailed calculation methods are shown in Yoon and Yoon . To calcThe incidence rate was primarily measured using NHI claims data, which generated YLD values. Mortality and fatality rates were measured using Statistics Korea's statistical yearbook for the cause of death. To measure YLL, the mortality rate and expected life expectancy for TB patients by age, gender, and year were measured using the Korean life table and the statistical yearbook . Cause-sThe economic burden was measured as the sum of direct and indirect costs for TB patients. For this calculation, NHI claims data and Korean health panel data were used. Direct costs were classified as medical and non-medical costs. In medical costs, insured medical costs include the total inpatient/outpatient medical and drug expenses, while uninsured medical costs include costs of service not covered by insurance. Furthermore, non-medical costs include transportation and caregiver costs .Indirect cost refers to productivity loss due to morbidity and premature mortality. Productivity loss due to morbidity includes the loss of time in outpatient visits and hospitalization, as well as loss of the average daily income. Subsequently, productivity loss due to premature mortality involves the number of deaths, life expectancy, and average annual income. Indirect cost also includes the loss of productivity due to use of hospital medical and loss of future income as a result of early death.Indirect cost loss appears to be a social loss, but the Health Insurance Review & Assessment Service (HIRA) does not generally implement policies that take into account indirect costs, and therefore the impact of TB is underestimated. With similar policies, in the case of rare diseases, medical institutions that can be diagnosed as well as treated continuously are concentrated in the metropolitan area, thus, patients living in rural areas were burdened with travel costs. The government is helping people with rare diseases to manage them by consulting, diagnosing and supporting patients with such diseases in the community. It is creating an education and request system through the establishment of a network of local medical services .This study was approved by the Institutional Review Board of National Evidence-based Healthcare Collaborating Agency . Informed consent was not required because public data from the NHIS database of de-identity were used.According to the data on DS-TB patients in the year 2017, 59.5% of this patient population was male, while 40.1% was female, and the average age was 59.6 years. In terms of income quintile, medical aid was 9.9%, and regional/workplace subscribers accounted for the majority 90.1%. In the MDR-TB group, 65.1% were men and 34.9% were women, with an average age of 53.5 years. In the income quintile, medical aid was 12.1%, and regional/workplace subscribers accounted for 87.9% .In DALYs per 100,000 people with DS-TB, the values were higher, as the data of an older population was considered. In terms of total DALYs for men and women, it was found to have decreased by 29% from 56 in 2014 to 40 in 2017. Overall, men with DS-TB had higher DALYs than their female counterparts. For most men with MDR-TB in all age groups, DALYs corresponded to 1 in 2017, a slight decrease from 2014. Contrary to the case of men, in women, DALYs appeared to be inconsistent across all age groups. The overall DALYs for men and women were found to have decreased by 33% from 3 in 2014 to 2 in 2017. Overall, no significant gender difference was found in DALYs for MDR-TB patients .The total cost of DS-TB in 2017 was the highest among men in their 40s and 50s. This parameter was relatively similar among women for all age groups. The total cost for both men and women experienced a 19% decrease from $143.89 million in 2014 to $116.62 million in 2017. MDR-TB was high for men in their 40s and older for women in their 70s and older. The total cost for men and women showed a slight decrease from $413.44 million in 2014 to $408.14 million in 2017. The decrease in death cost as part of indirect costs is believed to be the reason for this decline .In total cost by category, the direct cost of DS-TB in 2017 was $22.51 million, and the indirect cost was $94.11 million. This gave a total cost of $116.62 million. Medical costs account for a high percentage of direct costs, and productivity loss due to premature mortality account for 96% of indirect costs. From 2014 to 2017, the total cost showed a decreasing trend, which was estimated to be due to a decrease in death cost. The direct cost of MDR-TB in 2017 was $360.72 million and the indirect cost was $47.42 million, which gives the total cost of $408.14 million. Hospitalization cost constituted 60% of the direct cost, outpatient cost 31%, and care cost 8%. Productivity loss due to morbidity accounted for 96% of the indirect costs, which contradicts the finding for DS-TB. Compared to 2014, the total cost decreased slightly in 2017, which is estimated to be a result of a decrease in disease costs as part of indirect costs .This study measured the change in the status of TB burden for DS-TB and MDR-TB in Korea from 2014 to 2017 and evaluated the disease burden for each age group and gender. DS-TB-related DALYs per 100,000 people were found to have decreased over the study period . Subsequently, DALYs per 100,000 people for MDR-TB slightly decreased and then sustained during the observed period . The economic burden of DS-TB consistently reduced from 2014 to 2017, while that of MDR-TB fluctuated each year with no specific trend.DALYs per 100,000 people showed a decreasing trend for DS-TB and MDR-TB. In 2017, the DALYs per 100,000 people were 40 for DS-TB and 2 for MDR-TB. By gender, DALYs per 100,000 people was higher in men than those in women with DS-TB , but for MDR-TB, they were no significant difference between men and women . Previous study also showed a trend of decreasing DALYs per 100,000 people of general TB from 2014 to 2017, with 63 DALYs per 100,000 people for men and 36 DALYs per 100,000 people for women in 2017 . By age,In a 2015 study of economic burden, the total cost of TB was found to be $616.80 million, with a direct cost of $199.40 million and an indirect cost of $417.40 million . In thisAccording to the GBD study, the total DALYs for infectious diseases in 2017 was 6.5% of that of TB for all age groups, which is higher for patients aged 65 or above at 17% . StatistMDR-TB is a central issue in TB management due to its low treatment success rate, high mortality rate, and high disease burden. To increase the treatment success rate of MDR-TB, early diagnosis, appropriate treatment, and effective patient management must be accomplished by the national TB management system; there must be an efficient workforce capable of integrating and managing these aspects with the larger system . MDR-TB Meanwhile, the influx of foreign TB, especially MDR-TB patients, is increasing due to an increase in the inflow of people in Korea from high-risk countries for TB on the account of employment, education, and migration . FurtherOne study limitation was that fewer MDR-TB deaths were selected due to limitations in data sources. This is because MDR-TB deaths in the current year were analyzed in the study. Consequently, the YLL of MDR-TB was lower than that of DS-TB. However, these limited data alone confirmed that the DALYs of MDR-TB was low. In future studies, it will be necessary to select the data source to include a wide range of MDR-TB patients as subjects.Despite these limitations, the significance of this study is that it can be used as the basis for understanding the disease burden of TB by comparing DALYs and economic burden according to the presence or absence of drug resistance with the use of integrated data. Due to limitations in data sources and difficulties in selecting MDR-TB subjects, the current research on health outcomes using standardized methodologies is insufficient. However, this paper can be used as a reference study.Through this study, DALYs and economic burden on patients were compared for the study period of 2014\u20132017 according to TB resistance (DS-TB and MDR-TB). It was found that the total cost for DS-TB decreased and MDR-TB caused a huge social burden. Although the number of MDR-TB patients was fewer than that of DS-TB, the cost burden was extremely high for MDR-TB. In society, the aging population is rapidly increasing. Therefore, the incidence and spread of TB in all areas of society can be suppressed through intensive management of MDR-TB in the older population.To reduce the threat of early community transmission and resistance development for TB, public-private mixed collaboration for TB management was launched in 2009 . The maiIn particular, national-level MDR-TB patient management and support are required to reduce the number of TB deaths. While implementing a comprehensive TB management plan in Korea, the allocated budget should be increased based on the priorities. We hope that the national TB management plan will soon be efficiently implemented given the fact that the country's infectious disease management system is currently prioritizing the management of the COVID-19 over other diseases.https://nhiss.nhis.or.kr/bd/ay/bdaya001iv.do. This study used the National Health Information Database (NHIS-2019-1-662) of the National Health Insurance Service (NHIS).Publicly available datasets were analyzed in this study. This data can be found here: The studies involving human participants were reviewed and approved by National Evidence-Based Healthcare Collaborating Agency. Written informed consent from the participants' legal guardian/next of kin was not required to participate in this study in accordance with the national legislation and the institutional requirements.SL, MJK, and I-HO: conceptualization, data curation, and writing\u2014original draft. SL and I-HO: writing\u2014review and editing. SHL, H-YK, H-SK, and I-HO: supervison. All authors contributed and approved the article.This study was financially supported by the National Evidence-based Healthcare Collaborating Agency, funded by the Ministry of Health and Welfare , and by an Intramural Research grant from the Korean National Tuberculosis Association.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Uveitis, a sight-threatening ocular inflammatory state, is associated with autoimmune diseases and systemic inflammation. This prolonged systemic inflammation may cause plaque formation in coronary arteries, subsequently resulting in acute coronary syndrome (ACS).This retrospective, population-based study (15-year period) used the Longitudinal Health Insurance Database based on the National Health Insurance Research Database in Taiwan. Chi-square and Student\u2019s t-tests were used to examine differences between the study and comparison cohorts for categorical and continuous variables, respectively. Fine and Gray\u2019s competing risk model was used to determine the hazard ratio of the risk of ACS. Furthermore, the cumulative risk of ACS was determined using Kaplan-Meier analysis.A total of 1,111 patients with AS and uveitis were enrolled in this study cohort, and 4,444 patients with AS without uveitis were enrolled in the comparison cohort. After adjustment for age, sex, and comorbidities, patients with AS and uveitis demonstrated an increased risk of ACS compared to those without uveitis . In addition, Kaplan-Meier analysis revealed that patients with AS and uveitis had a significantly higher risk of ACS than those without uveitis (p<0.001). Age, diabetes mellitus, hypertension, hyperlipidemia, chronic obstructive pulmonary disease, asthma, and systemic steroids were significant risk factors for ACS. Both anterior uveitis and posterior segment involvement were associated with an increased risk of ACS in patients with AS. All-cause mortality was higher in the uveitis group (9.81%) than in the non-uveitis group (8.10%) (p=0.015).Our analysis revealed that uveitis could potentially be a predictor of ACS in patients with AS. However, further prospective controlled studies are required to assess the association between uveitis and ACS in patients with AS. Ankylosing spondylitis (AS) is a chronic inflammatory arthritis that involves the axial skeleton, belonging to a group of diseases known as spondyloarthritis . UsuallyUveitis, or inflammation of the uvea, is the most frequent extra-articular manifestation of AS, followed by psoriasis and inflammatory bowel disease . Pro-infTaking into account the above-mentioned considerations, we designed a retrospective matched-cohort study that used claims data from the National Health Insurance Research Database (NHIRD), which encompasses a comprehensive longitudinal medical record from almost the entire population of Taiwan (23 million), to evaluate the association between patients with AS and uveitis and the risk of ACS.This study employed the Longitudinal Health Insurance Database (LHID) from the NHIRD to examine the association between uveitis in patients with AS and subsequent development of ACS in Taiwan over a 15-year period. The claims data contained medical records of outpatients and inpatients and were registered using the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) codes. This NHIRD included around 99% of 23 million Taiwanese citizens and represented the real-world data in Taiwan . PatientWe retrospectively conducted a matched-cohort study from January 1, 2000, to December 31, 2015. Patients were selected if they had at least one inpatient claim or more than three outpatient visits with the diagnosis of AS (ICD-9-CM code 720.0). The index date was defined as the date on which the patient was first diagnosed with AS. Patients diagnosed with AS or ACS before the index date were excluded to ensure that all ACS cases were newly diagnosed after AS. Patients aged 20 years and younger, of unknown sex, or without tracking follow-up were excluded. The study population was divided into uveitis (study cohort) and non-uveitis (comparison cohort) groups. The non-uveitis group was randomly matched fourfold with the uveitis group according to sex, age, and index year (under the same exclusion criteria). Furthermore, the uveitis group was divided into the anterior uveitis and posterior segment involvement groups. Patients were tracked until ACS onset or the end of the study period, whichever occurred first. The selection process is illustrated in Uveitis was the main independent variable of interest. The evaluated covariates included sex, age, diabetes mellitus (DM), hyperlipidemia, hypertension (HTN), asthma, and chronic obstructive pulmonary disease (COPD). The considered medication was systemic steroids. These factors were matched at baseline and considered confounders and adjusted for statistical analysis.All statistical analyses were performed using SPSS software version 22 . The differences between the study participants and the comparison cohort for categorical and continuous variables were assessed using the chi-square and t-tests, respectively. A Cox proportional hazards model with Fine and Gray\u2019s competing risk model was used to determine the hazard ratio of risk of ACS based on each variable. Survival analysis was performed using the Kaplan-Meier method with log-rank test. A two-tailed p value <0.05 was considered statistically significant.This study was approved by the Institutional Review Board of the Tri-Service General Hospital (TSGHIRB: B-110-41), and the need for individual written informed consent was waived. This study was conducted in accordance with the code of ethics of the World Medical Association (Declaration of Helsinki).As depicted in The results of the stratified analyses of the risk factors associated with the development of ACS are shown in The uveitis group was further grouped into anterior uveitis (n=394) and posterior uveitis (n=717) groups, as shown in In this 15-year population-based investigation on the correlation between development of ACS and uveitis in patients with AS, we found that patients with AS and uveitis showed a significantly increased risk of ACS compared to those without uveitis, even after adjusting for age, sex, and comorbidities. Similarly, Kaplan-Meier analysis revealed that the cumulative risk of ACS was significantly higher in the uveitis group than that in the non-uveitis group. Uveitis demonstrated a strong association with an increased risk of ACS in both sexes, all ages, and patients with AS with and without DM, hyperlipidemia, HTN, COPD, and asthma. In the subgroups of uveitis, both anterior uveitis and posterior segment involvement demonstrated an increased risk of ACS in patients with AS. This study identified uveitis as a risk factor for ACS in patients with AS and suggests that it may be a potential predictor of ACS in patients with AS.The prevalence of AS ranges from 9 to 30 per 10,000 people in the general population . AlthougPatients with AS tend to have a higher rate of comorbidities than the general population. The prevalence of HTN, DM, and asthma in patients with AS was 30.7%, 9.8%, and 2.2%, respectively, in the US , and theAging, hypertension, and diabetes mellitus are associated with the development of ACS , 28 and Uveitis is the most common extraarticular manifestation of AS .The mechIn addition to inflammation, AS has a strong genetic component , and HLAOur study reported a higher aHR for ACS in the posterior segment involvement group than in the anterior uveitis group Table\u00a04.Clinically, the treatment of non-infectious uveitis (NIU) included topical steroids or systemic steroids, disease-modifying antirheumatic drugs , and biologic agents \u201347. UveiA strength of this study is the large sample size within a comprehensive database spanning over a 15-year period. This not only provides high statistical power that can best reflect real-world situations, but also minimizes selection bias. In addition, this is the first population-based study to investigate uveitis as a risk factor for acute coronary syndrome in patients with AS. Furthermore, this study matched comorbidities at baseline, which could provide more reliable results for adjusting these variables.This study had several limitations that should be addressed. First, misclassifications of the subtype of uveitis and cardiovascular outcomes may be inevitable; however, the National Health Administration in Taiwan checks charts to ensure that patients have appropriate claims and treatment. Second, the LHID only included the population of Taiwan, thus, this result needed to be validated and extrapolated by a further study in a much bigger group. Third, due to that our research was a retrospective study by analyzing the fully anonymized database, we could not obtain other confounding variables . Hence, the interferences of these confounding variables needed a further prospective study to determine these.In conclusion, this study demonstrated that uveitis is an independent risk factor for ACS in patients with AS. In patients with AS and uveitis, a trend of specific comorbidities showed a higher risk of ACS. The results of this study will allow ophthalmologists, rheumatologists, or any medical doctor to be aware of the risk of ACS when monitoring patients with AS and uveitis.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by Institutional Review Board of the Tri-Service General Hospital (TSGHIRB: B-110-41). Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.KF, W-CC, Y-HC, J-TC, and C-LC: study design and manuscript writing. W-CC, C-AS, C-HC, and C-LC: data extracting and statistical analysis. K-MF, Y-HC, C-AS, J-TC, and C-LC: data checking. All authors contributed to the article and approved submitted version.This study was funded by the Tri-Service General Hospital Research Foundation (TSGH-D-110112 and VTA111-V1-1-2) and by the Ministry of Science and Technology (MOST 107-2314-B-016-030). The sponsors have no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The Internet of Things (IoT) is a widely used technology in automated network systems across the world. The impact of the IoT on different industries has occurred in recent years. Many IoT nodes collect, store, and process personal data, which is an ideal target for attackers. Several researchers have worked on this problem and have presented many intrusion detection systems (IDSs). The existing system has difficulties in improving performance and identifying subcategories of cyberattacks. This paper proposes a deep-convolutional-neural-network (DCNN)-based IDS. A DCNN consists of two convolutional layers and three fully connected dense layers. The proposed model aims to improve performance and reduce computational power. Experiments were conducted utilizing the IoTID20 dataset. The performance analysis of the proposed model was carried out with several metrics, such as accuracy, precision, recall, and F1-score. A number of optimization techniques were applied to the proposed model in which Adam, AdaMax, and Nadam performance was optimum. In addition, the proposed model was compared with various advanced deep learning (DL) and traditional machine learning (ML) techniques. All experimental analysis indicates that the accuracy of the proposed approach is high and more robust than existing DL-based algorithms. The IoT foresees the networking of a wide range of smart things in our environment that are capable of accumulating, processing, and communicating data . The IoTMany IoT devices capture, store, and process personal data, making them a feasible target for assailants because of their distributed structure and openness . The effThe existing system has difficulties in improving performance and identifying subcategories of cyberattacks. This paper proposes a DCNN followed by a deep-neural-networks (DNN)-based IDS. The primary advantage of a DCNN is its ability to exploit the correlation between features . A DCNN We proposed a DCNN technique for malicious activity identification in IoT networks.We improved performance and reduced the computational power of an IDS for low-power IoT devices in the network.We identified the subcategory of cyberattacks in the IoT networks.We compared the proposed scheme with other DL and traditional ML techniques.The remainder of the article is organized as follows. Security is an essential part of an IoT network for stability, reliability, and safe communication. Several researchers have proposed different techniques for the detection of malicious attacks in IoT networks. Basati et al. presenteFatani et al. introducA single hidden layer feedforward neural network (SLFN) method was introduced by Qaddoura et al. for maliThe existing systems cannot identify the subcategories of multi-class attacks in the network. In addition, for binary and multi-class detection, the performance of the existing system can be improved. A comparison of the related work is given in This section provides a detailed explanation of the utilized dataset, preprocessing approaches, the proposed deep convolutional neural network (DCNN), and evaluation metrics.The IoTID20 dataset was developed to identify cyberattacks in IoT networks. This dataset was generated through home-connected smart devices using SKT NGU and EZVIZ Wi-Fi cameras . The maiData preprocessing is an essential step for ML/DL methods. Preprocessing converts data into a suitable format for any neural network. This section consists of cleaning, label encoding, feature engineering, normalization, and data splitting.A dataset must be verified for empty and undefined instances before training a model. In this experiment, the Python built-in library (Pandas) was used to validate the dataset. The utilized IoTID20 dataset has some missing values. To clean the dataset, we removed all missing value instances.Label encoding is a well-known encoding approach for dealing with categorical values. It assigns a unique numeric value to each categorical value. For ML algorithms and DL neural networks to operate, the input and output values must be integers. The utilized dataset has some categorical features. Each categorical feature has several categories for which one-hot encoding requires greater memory and more time . In thisEach dataset contains its own set of features. If a dataset contains multiple features as well as certain insignificant features that have no impact on the output label, we must eliminate those features from the dataset because they lead to overfitting and underfitting, which significantly influence the executing time and performance of the classifier. In this study, the filter approach was used. In filtering features, the extra tree classifier (ETC) technique was applied. This method calculates the impact of each feature on the output label. The utilized dataset has 83 features. We select all the features greater than 0.001 for information gain. After applying the feature filtering approach, 62 features were selected.Normalization is a method commonly used in the preprocessing of data for ML/DL algorithms. The purpose of normalization is to convert the numeric column values in a dataset to a common scale while maintaining variations in value ranges. Each feature of the IoTID20 dataset has different values. Some feature values are in the thousands, and some have negative values that reduce the model performance. To solve this problem, the data are normalized between 0 and 1 via min\u2013max method, as represented by Equation which is input in the first dense layers. The output of the first dense layer is which is given as input in the second dense layer. The second dense layer produces output which is input in the last dense layer. The ReLU activation function is used in dense layers. The last dense layer produces output results in which sigmoid function for binary classification and softmax function for multi-class classification are used, respectively. Sigmoid and softmax are demonstrated in Equations (6) and (7).The evaluation of the DCNN approach was carried out with accuracy, precision, recall, and F1-score. We start by explaining these four parameters, true positive (TP), false negative (FN), false positive (FP), and true negative (TN), which are used to compute the evaluation metrics such as accuracy, precision, recall, and the F1-score. TP refers to the number of instances that have been correctly identified as normal. The number of instances that misclassify normal data as an attack is known as the FN. FP represents the number of malicious instances that are wrongly classified as normal. TN represents the number of instances that are classified correctly as malicious. All of these evaluation metrics were calculated by using Equations (8)\u2013(11).Experiments on the DCNN model were conducted with the HP ProBook G5 8th generation laptop. This laptop contains 24 GB ram and an Intel Core i5 processor. In software specifications, we used Windows 11 Pro, Python 3.8.5, Tensorflow, and Keras library.This section provides a detailed evaluation of the proposed model. The proposed DCNN model was evaluated on the IoTID20 dataset. The performance of the DCNN was tested for binary, multi-class categories, and multi-class subcategories classifications. This section presents a comparison of convolutional layers followed by dense layers for multi-class categories and multi-class subcategories. The same comparison was performed for famous optimizers. The optimal solutions were selected from the comparison and compared with other ML/DL models.The CNN algorithm consists of convolutional layers, pooling layers, and fully connected layers. This experiment was conducted for one and two convolutional layers, followed by fully connected dense 1\u20135 layers. These experiments were conducted for the multi-class category and subcategory classification. A detailed comparison is given in An optimizer is a function used to update the neural network weights and learning rates. It helps to reduce the loss and improve the performance of the model ,32. FamoIn this study, we propose a DCNN architecture for malicious activities identification in IoT networks. For DCNN, the above results show that the optimal solution for the IoTID20 dataset is two convolutional layers, followed by three dense layers. In addition, from the above results, we selected the top three optimizers for this experiment. This section provides a detailed classification of binary-class, multi-class category, and multi-class subcategories for batch sizes 32, 64, 128, and 256.The performance of the proposed approach was tested for a binary-class scenario. The DCNN model was trained with the IoTID20 dataset for 50 epochs, and the binary cross-entropy function was used to calculate the loss. In the first step, the proposed DCNN performance for the Adam optimizer is compared in the bar graphs in In this stage, the performance of the proposed study was evaluated for a multi-class category classification scenario. The DCNN model was trained with the IoTID20 dataset for 50 epochs, and a sparse categorical cross-entropy function was used to calculate the loss. As noted previously, for the binary-class studies, an Adam optimizer was chosen at the initial stage. The proposed DCNN performance for the Adam optimizer is compared in the bar graphs in In the final stage, the performance of the proposed study was evaluated for multi-class subcategory classification scenarios. The DCNN model was trained with the IoTID20 dataset for 100 epochs, and a sparse categorical cross-entropy function was used to calculate the loss. As noted previously, for the binary and multi-class category studies, an Adam optimizer was chosen at the initial stage. The proposed DCNN performance for the Adam optimizer is compared in the bar graphs in The performance of the proposed DCNN was analyzed for binary, multi-class category, and multi-class subcategory classification. The results presented earlier show a comparison of optimizers and batch sizes. Based on the performance analysis of the proposed model for binary class, the Nadam optimizer with a batch size of 128 performs better than the others. Similarly, in the performance analysis of the proposed model for the multi-class category and subcategory classification, the Adam optimizer with a batch size of 32 performs better than others. For testing the performance of the proposed model, k-fold cross-validation was also used, where the \u201ck\u201d value is 7. The results of the k-fold cross-validation are approximately equivalent.The performance of the proposed DCNN was compared with other DL and traditional ML methods to evaluate its efficacy. LSTM, gated recurrent unit (GRU), deep neural network (DNN), deep belief network (DBN), deep autoencoder (DAE), and multilayer perceptron (MLP) are examples of DL methods. Decision tree (DT), logistic regression (LR), naive Bayes (NB), support vector machine (SVM), and k-nearest neighbors (KNN) are all examples of traditional ML methods. All of these methods were implemented in the same environment for an accurate performance comparison. The preprocessing steps were the same for all models, including the proposed model. We split the dataset into 80% train and 20% test sets. For all of the DL algorithms, we used Adam optimizer and default batch size 32. The optimal solution of each model was used for the comparison. The hidden layers used in LSTM, GRU, DNN, DBN, AE, and MLP are 3, 3, 4, 4, 6, and 10, respectively. The number of training epochs for all these models was the same as the proposed model. A detailed analysis for binary-class category, multi-class category, and subcategory classifications is shown in For optimal performance, each DL model requires multiple layers that maximize computational power. The proposed DCNN model improves the performance and also reduces computational power as it narrows to specific features, compared to other ML and DL models. Comparing the performance of the proposed DCNN with other ML and DL models shows the optimal results.This study presents a new DCNN-based DL model and feature engineering method for malicious attack detection in IoT networks. The objective was to improve performance and reduce computational power. The proposed DCNN model successfully improves performance and reduces computational power. It is useful for low-power IoT network devices. The IoTID20 dataset was used to analyze the performance of the proposed DCNN model. The proposed model was evaluated for binary, multi-class category, and subcategory classifications. Experiments were performed for different layers of the CNN algorithm, and an optimal solution was selected. The proposed model was evaluated in-depth with Adam, Nadam, and AdaMax optimizers. The Nadam optimizer peformance was optimum for binary, multi-class category, and multi-class subcategory with 128, 32, and 64 batch sizes, respectively. The proposed model was also compared with state-of-the-art DL techniques and other traditional ML algorithms for a broader view in terms of efficacy, robustness, etc. The experimental analysis indicates that the proposed approach obtained optimum results when compared through accuracy, precision, recall, and F1-score parameters."} +{"text": "To assess whether dietary knowledge of Chinese children and adolescents and their mothers was associated with childhood and adolescent overweight and obesity.This cross-sectional study obtained data from the China Health and Nutrition Survey (CHNS) between 2004 and 2015. Dietary knowledge of children and adolescents and their mothers was assessed by asking questions and statements on diets, and clustered by K-means clustering. Body mass index (BMI) and waist circumference (WC) were used to evaluate overweight and obesity among children and adolescents. The association of dietary knowledge with childhood and adolescent overweight and obesity was evaluated by multivariate regression analysis, with odds ratios (ORs) and 95% confidence intervals (CIs) calculated.P = 0.002), and WC-defined obesity than those with high dietary knowledge. Compared with high dietary knowledge in mothers, low dietary knowledge was associated with significantly elevated risks of BMI-defined overweight or obesity , and WC-defined obesity . Furthermore, significantly increased odds of BMI-defined overweight or obesity and WC-defined non-obesity in children and adolescents were related to low dietary knowledge versus high dietary knowledge of children and adolescents , while there was no association of BMI-defined non-overweight and non-obesity and WC-defined obesity with dietary knowledge among children and adolescents . Additionally, no association was found between dietary knowledge of mothers and BMI-defined overweight or obesity and WC-defined non-obesity among children and adolescents , while low dietary knowledge of mothers was associated with increased odds of BMI-defined non-overweight and non-obesity and WC-defined obesity in children and adolescents .A total of 2,338 children and adolescents were included. Children and adolescents with low dietary knowledge were demonstrated to have significantly higher risks of BMI-defined overweight or obesity (OR = 1.66, 95%CI = 1.21\u20132.28, Dietary knowledge of children and adolescents and their mothers was associated with childhood and adolescent overweight and obesity. Dietary knowledge of children and adolescents negatively related to the risk of BMI-defined overweight or obesity, and dietary knowledge of mothers to odds of WC-defined obesity. Overweight and obesity in children and adolescents has become one of the most serious public health problems of the 21st century, with significantly increasing worldwide prevalence . In 2017et al. , and the Mann-Whitney U rank sum test was applied for intergroup comparison; enumeration data were shown by the number of cases and constituent ratio [n (%)], and the Chi-square test or Fisher\u2019s exact test was employed for comparison between groups. Missing data were subjected to multiple imputation (R: mice), and sensitivity analysis was performed via difference analysis on the data before and after imputation. The number of clusters was determined, and K-means clustering was used to cluster dietary knowledge of children and adolescents, and their mothers, respectively. The screened confounding factors were put into multivariate models as covariates, along with dietary knowledge of children and adolescents. Multivariate regression analysis was conducted with overweight and obesity defined by BMI and WC as the outcome variables, with odds ratios (ORs) and 95% confidence intervals (CIs) calculated. Model 1 was a univariate model, model 2 was adjusted for age, gender, and mother\u2019s education level. Model 3 was adjusted for age, gender, mother\u2019s education level, geographic location (area and province), SBP, and DBP. In model 3, television time in a week was additionally corrected for when obesity in children and adolescents was evaluated via WC. All statistical tests were two-sided, and P < 0.05 was considered statistically significant. Statistical analysis was performed using SAS 9.4 and R 4.20 software.Normally distributed measurement data were described as mean \u00b1 standard deviation (Mean \u00b1 SD), and the independent samples t-test was used for comparison between groups; non-normal data were expressed as median and quartile [M (QP < 0.05). For the non-obese and obese groups by WC, significant differences existed in SBP, DBP, area, province, hip circumference, and mother\u2019s education level . Besides, two clusters were established for dietary knowledge of children and adolescents or drink (91.87%), and lived in rural areas (66.25%). Most mothers had a lower middle school degree (40.98%). The flow chart of study population selection is shown in lescents , and twolescents .P = 0.001), and obesity defined by WC among children and adolescents. Similarly, dietary knowledge of mothers was significantly associated with overweight or obesity defined by BMI , and obesity defined by WC in children and adolescents. Then following covariate adjustment in model 3, children and adolescents with low dietary knowledge was demonstrated to have significantly higher risks of BMI-defined overweight or obesity , and WC-defined obesity than those with high dietary knowledge. Compared with high dietary knowledge in mothers, low dietary knowledge was associated with significantly elevated risks of BMI-defined overweight or obesity , and WC-defined obesity .P < 0.0001). Dietary knowledge of children and adolescents was inversely associated with odds of overweight and obesity in children and adolescents who was overweight or obese defined by BMI and obese defined by WC . Significantly increased odds of BMI-defined overweight or obesity and WC-defined non-obesity in children and adolescents were found to be related to low dietary knowledge versus high dietary knowledge of children and adolescents , while there was no association of BMI-defined non-overweight and non-obesity and WC-defined obesity with dietary knowledge among children and adolescents . Additionally, low dietary knowledge of mothers was significantly associated with BMI-defined overweight or obesity or WC-defined obesity in children and adolescents . A significantly greater risk of BMI-defined overweight or obesity and WC-defined obesity in children and adolescents was in relation to low dietary knowledge rather than high dietary knowledge in mothers . No association was identified between dietary knowledge of mothers and BMI-defined overweight or obesity and WC-defined non-obesity among children and adolescents , while low dietary knowledge of mothers was associated with increased odds of BMI-defined non-overweight and non-obesity and WC-defined obesity in children and adolescents .Further, we explored the association between dietary knowledge and overweight and obesity in children and adolescents defined by BMI and WC together. As presented in In order to investigate the relationship of dietary knowledge and overweight and obesity in Chinese children and adolescents, this study applied data from the CHNS and utilized BMI and WC to assess overweight and obesity. It was revealed that dietary knowledge of children and adolescents and their mothers was associated with their overweight and obesity. Further, low dietary knowledge of children and adolescents related to an elevated risk of BMI-defined overweight or obesity, and dietary knowledge of mothers was negatively associated with odds of WC-defined obesity.Studies using anthropometric measurements have found that nutrition knowledge is related to BMI and WC, which are the indicators of obesity and its comorbidities, such as cardiovascular diseases , 27. TheDietary knowledge was revealed here to correlate with overweight and obesity in children and adolescents. Bonaccio et al. proposed that individuals with nutrition knowledge showed greater compliance with the Mediterranean diet and lower obesity prevalence . AnotherWe assumed that the dietary knowledge level affects diet quality and physical activities, which influence childhood and adolescent overweight and obesity , 32. A pThis study first assessed the association of dietary knowledge of children and adolescents and their mothers with childhood and adolescent overweight and obesity, using the nationally representative CHNS data. K-means clustering was used to cluster the dietary knowledge of children and adolescents, and their mothers into two different groups . Some limitations should be noted when interpreting findings. First, a cause-and-effect relationship could not be determined in this cross-sectional study. Second, stability of weight status was not measured in this study, and the reported weight was assumed to be the long-term result of eating self-regulation. Third, data on dietary knowledge were self-reported, which may over- or under-estimate actual weight status. Fourth, variables such as physical activity were not included in the analysis due to limited information from the CHNS.Dietary knowledge of children and adolescents and their mothers was associated with childhood and adolescent overweight and obesity. Further, low dietary knowledge of children and adolescents related to an elevated risk of BMI-defined overweight or obesity, and dietary knowledge of mothers was negatively associated with odds of WC-defined obesity. Additional studies are required to confirm our findings and to clarify the causal relationship between dietary knowledge and overweight and obesity among children and adolescents.S1 Checklist(DOCX)Click here for additional data file."} +{"text": "Accounting for over 70% of the Earth\u2019s surface, the oceans have provided enriched resources and abundant space for humankind. Maritime exploration and economic development have placed tremendous challenges for contemporary technologies. Robotic systems can significantly reduce risks, increase efficiency, and extend the range of explorations in harsh marine environments. For long-term operations in the oceans, energy sustainability is a universal research theme in marine robotics, serving as foundations for locomotion, sensing, communication, control, and other related topics.This Research Topic aims to showcase a variety of perspectives in energy sustainability so as to inspire and promote cutting edge innovations for marine robots with long-term operation capabilities. It presents a comprehensive review and original research articles in topics including efficient utilization of energies, re-charging, path planning, and state estimation. Both marine surface and underwater robots are investigated. Specifically, the following papers are included:Sun et al. provide a comprehensive review from the energy perspectives on sailing robots for long-term operation goals. Prototypes and products developed in academia, industry, competition, and open community are categorized from aspects of sail types, hull types, electricity harvesting components, etc. Continuous progress from main contributing teams are elaborated for tracking of their R&D works. Actuations, energy harvesting, and energy management are analyzed. Various insights, e.g., motorized propellers, wave-based propulsion, balanced rudders, solar panel angles, multimode energy/E-saving management strategies, and so forth, can be used in sailboat design, control, and energy management.Negreiros et al. present two robotic sailboats, named N-Boat and F-Boat, aiming for sustainable solutions to monitor the ocean. Energy consumption estimation and management is studied. A Restricted Boltzmann Machine is utilized to predict the sailboat\u2019s energy consumption in a 24-h range, so as to find the most efficient way for solar panel energy harvesting. The forecasted trend of power consumption with and without solar panel fits the real data well.Yang et al. present an unmanned sailboat platform with generic and flexible features, aiming towards the World Robotic Sailing Championship. The system is based on a 1-m class RC sailboat, with added low-cost open-source hardware module, e.g., Pixhawk, Arduino, GPS, wind direction sensor, wireless 433\u00a0MHz telegram, etc. Line-of-sight guidance strategy and PID control enables the robot to sail at way-points defined by users. Missions of fleet racing, station keeping, and area scanning validates the system and algorithms.Gao et al. propose a novel design on the oscillating foil to improve the wave-energy-to-propulsion conversion efficiency. The main concept is that it adopts an asymmetric foil structure design and asymmetric upper and lower oscillating limit angle, which breaking through the traditional idea that the design should be symmetric for wave energy harvesting. This paper includes both the analytical and simulation analysis to evaluate the improvement of the propulsion. The proposed design has been realized and applied on a simplified wave glider, and several open sea experiments have shown the effectiveness of the proposed design.Page et al. present an integrated navigation algorithm for reliable underwater docking and recharging of propeller-driven autonomous underwater vehicles (AUVs), so as to enhance their energy sustainability. It dynamically re-plans efficient Dubins paths from the current position towards a terminal homing location. Then the system controls an AUV by integral line of sight to follow the path. The AUV tracks the light from the hand-off location into the dock. Experiments based on two AUVs have validated the approach in lakes and pools. This approach is promising for persistent undersea operations.Alam et al. explore the feedback planning approach, which is a function over belief space to produce an action to reach the goal belief state, aiming for energy-awareness in long-range AUVs. It attempts to overcome challenges of uncertainties on motion and sensors in the underwater environment. With a water flow pattern generated from Regional Oceanic Modeling System (ROMS) known a-priori, passive actions of drift are performed to save energy, while paths towards goal states are generated in different water layers through a variety of simulations.Ismail et al. present a new form of data assimilation with a smooth variable structure filter with an application for estimating the missing states of rivers. The principal advantages of this work are 1) overcoming the missing observation and 2) improving the estimation performance for Lagrangian marine drifters. The measurements from the drifters are used in the estimation of the flow, stage, and cross section that are later used to evaluate the velocity of the sixth Lagrangian marine sensor. The convergence analysis is discussed. A comparison between the proposed method with the extended Kalman Filter shows the positive effect of the proposal.The Guest Editors would like to thank all the authors and reviewers for their work and devotion to the Research Topic, and hope that it can inspire further research in energy sustainability for marine robotics."} +{"text": "M. Lesley Wiseman-Orr was not included as an author in the published article. The published research was divided into two parts \u2014 an \u201calert\u201d development phase for an existing health-related quality of life (HRQL) tool and some 5 years later the reporting of use of the alert in a large cohort of dogs, which was the main focus of the paper. Although Dr. Wiseman-Orr was not involved in the second part of the study, she was involved in the conceptualization of the study, and in the development of the alert and the app. The original authors now appreciate that omitting her from the author list was a most unfortunate oversight on their part and wish to remedy that if possible.The corrected Author Contributions Statement appears below.Conceptualization: MW-O, JR, VD, and ES. Provision of data: JR and AW. Statistical analysis: VD and ES. Algorithm: VD, MW-O, and JR; App development: JR and MW-O. Writing, review, and editing: JR, VD, ES, MW-O, and AW. All authors contributed to the article and approved the submitted version.The authors apologize for these errors and state that they do not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "In the original article, there was an error in the Funding statement as published. The funder name was incorrectly written as \u201cMinistry of Science and Technology, Taiwan, R.O.C.\u201d The corrected Funding statement is shown below.This research was funded by the National Social Science Foundation of China (Grant No. BCA200093) and Priority Academic Program Development of Jiangsu Higher Education Institutions. In addition, this study was supported by the Ministry of Science and Technology, Taiwan, under grants MOST 109-2511-H-019-004-MY2 and MOST 109-2511-H-019-001.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Correction to: BMC Health Serv Res 21, 1053 (2021)https://doi.org/10.1186/s12913-021-07069-wFollowing publication of the original article , an errobold typeface.The article title should read, and the change has been highlighted in care for the Migrant Parents in China-- evidence from the China migrants dynamic survey.Does integrated medical insurance system alleviate the difficulty of using cross-region health In addition, the city and country of the author affiliation 3 are incorrect. The updated author affiliation 3 should be:University of International Business and Economics, Institute of International Economy, Beijing, China.The original article has been"} +{"text": "The Journal of Nursing Research, four articles address issues that influence professional identity formation in the nursing profession. Their topics include the effects of education on professional quality of life and health among nurses, the effects of film-based nursing education in developing professional nursing identity among nursing students, the impact of organizational support on practice outcomes in nurse practitioners, and the use of a function-focused interdisciplinary communication framework in a nursing home setting.The nursing profession is evolving rapidly to meet the challenges and opportunities presented by the constant, technology-driven changes in healthcare delivery and the ever-changing landscape of healthcare needs. In this issue of Evidence-based nursing interventions are paramount to the successful development and adoption of evidence-based practices in nursing. Two articles in this issue provide evidence regarding the efficacy of nursing interventions on different groups of nursing clients, including the efficacy of machine-based hand massage in patients awaiting outpatient surgery and the efficacy of auricula acupressure in community-dwelling poor sleepers.Self-management is an important strategy for reducing chronic-disease-related symptom distress and illness burden. Also in this issue, the results of a survey querying the perceived effectiveness of pain self-management strategies in people with migraine headache are reported.While high-quality quantitative studies are needed for evidence-based nursing practice, the qualitative research approach is useful for exploring issues and phenomena that are poorly defined or understood. To this end, the components of empowerment in family caregivers of community-dwelling people with dementia were also explored in a qualitative research study included in this issue.Copyright \u00a9 2021 The Authors. Published by Wolters Kluwer Health, Inc. All rights reserved. This is an open access article distributed under the Creative Commons Attribution License 4.0 (CCBY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.ptsai@tmu.edu.twAddress correspondence to: Pei-Shan TSAI, PhD, RN, Professor, School of Nursing, College of Nursing, Taipei Medical University, No. 250, Wu Xing St., Taipei City 11031, Taiwan, ROC. Tel: +886-2-27361661 ext. 6321; E-mail: Cite this article as:The Journal of Nursing Research, 29(3), Article e147. https://doi.org/10.1097/jnr.0000000000000438Tsai, P.-S. (2021). The varied and multifaceted professional roles of today\u2019s nurses."} +{"text": "There are errors in the funding statement. The correct funding statement is as follows:https://www.ciencia.gob.es/) and the Academy of Finland Centre of Excellence in Experimental and Computational Biology (Suomen Akatemia) AA123456 (https://www.aka.fi/en/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.This study has been funded by the Spanish Ministry of Science PGC2018-096802-B-I00 ("} +{"text": "Reinhold Mankau (1928\u20132021)Reinhold Mankau passed away on Sunday, December 5, 2021, in Riverside, California. Known by his family, friends, and colleagues as \u2018Ron\u2019, he was a faculty member at the University of California Riverside for 33 years. He belonged to the first generation of scientists that gave the UCR Department of Nematology its stellar worldwide reputation.Heterodera trifolii. In 1954, he and the love of his life, Saroj, got married. A year later, both finished with their Ph.D. Ron received a Fulbright Research Fellowship at the Indian Agricultural Research Institute, New Delhi, India, where they lived from September 1956 to December 1957. A month later, he was appointed Assistant Nematologist at the University of California, Riverside, pursuing microbial biocontrol organisms.Ron was born in Chicago, Illinois, on July 22, 1928. He studied Biology from 1946 to 1948 at Wright Junior College, Chicago. The following year, Ron enrolled at the University of Illinois and received a bachelor\u2019s degree in 1951 and a Master of Science degree in 1952, both in Biology. As a Ph.D. student in Plant Pathology/Parasitology at the land-grant University of Illinois, Urbana, Plant Pathologist/Nematologist Dr. Maurice Linford was his major professor. Ron\u2019s thesis focused on host\u2013parasitic relationships of the clover cyst nematode, In 1964\u201365, he spent a sabbatical as a Fulbright Senior Postdoctoral Fellow at the Agricultural College and Research Institute, Coimbatore, in southern India. In 1972, he hired Diana Wall Freckman as a Research Nematologist in his lab which started her long and exceptional career. In 1973\u201374, Ron served as a United Nations Development Program consultant in a project designed to advance the postgraduate program in Plant Pathology/Nematology at the University of Agricultural Science, Hebbal, Mysore, India. Consequently, he was one of the foremost experts regarding plant-parasitic nematode problems and research in India.Ron\u2019s research on biology and epidemiology of parasites and predators of plant-parasitic and free-living nematodes earned him a worldwide reputation as an authority in biological nematode control and sustainable crop management. Ron knew his nematode-destroying fungi like no other. His lab studied physical and biological factors that influenced their reproduction, persistence, and efficacy. His mission-oriented goal was to find microorganisms that could be efficiently and economically useful as biological control agents.Meloidogyne spp.) in southern California. Gerald Thorne, one of the pioneers of US Nematology, had described the microorganism previously as the microsporidian parasite Duboscqia penetrans. But Ron Mankau and Jack Imbriani published in 1974 after extensive study of morphology, histochemistry, and life cycle in various nematodes that the organism was a prokaryote. Ron renamed it Bacillus penetrans (now Pasteuria penetrans). However, the true potential of this and similar bacteria occurred to him while working for the French Office of Scientific and Technical Research Overseas (ORSTOM) in Senegal in 1979. He discovered B. penetrans parasitizing root-knot nematodes in some grower fields that significantly mitigated crop damage by those otherwise devastating pathogens. Equally important was the discovery by his Ph.D. student Graham Stirling of Dactylella oviparasitica , a hyperparasite of root-knot nematodes. They demonstrated the importance of the fungus in the natural suppression of M. incognita populations in some California peach orchards.Ron also was a trailblazer in researching nematode-suppressive soils. In early 1970, he discovered a bacterium in mountainous areas of California that attached to the surface of nematodes. He found similar organisms associated with greenhouse and field populations of root-knot nematodes (Ron taught the formal class Nematode Diseases of Plants for many years together with his Department of Nematology colleagues Ivan Thomason, Seymour Van Gundy, and Howard Ferris. He served on the Steering Committee for UCR\u2019s Pest Management M.S. Program. He trained Ph.D. students, post-docs and inspired many more worldwide. On his and Saroj\u2019s travels, he was often invited to give scientific seminars which he presented fluently in German, French, or Spanish. Ron was promoted to the rank of Full Professor in 1976 and retired in 1990. In Varadero, Cuba, at one of the last scientific meetings he attended, the Organization of Nematologists of Tropical America honored him with a citation of special recognition for his long-standing contribution and support of ONTA since the inception of the organization.In retirement, he and his wife Saroj, a California State University Emeritus Professor in Biology, spent most of the year in southern Baja. Throughout their 67 years of marriage, they enjoyed traveling together, making friends on all continents, and visiting more than 100 countries around the globe. Ron was humble and personable and remained involved with the Nematology Department until shortly before his death. He will be remembered fondly by many."} +{"text": "Background: The Chinese Neonatal Network (CHNN) is a nationwide neonatal network that aims to improve clinical neonatal care quality and short- and long-term health outcomes of infants. This study aims to assess the quality of the Chinese Neonatal Network database by conducting an internal audit of data extraction.Methods: A data audit was performed by independently replicating the data collection and entry process in all 58 tertiary neonatal intensive care units (NICU) participating in the CHNN. Eighty-eight data elements selected for re-abstraction were classified into three categories , and agreement rates for original and re-abstracted data were predefined. Three to five records were randomly selected at each site for re-abstraction, including one short- (0\u20137 days), two medium- (8\u201328 days), and two long-stay (more than 28 days) cases. Agreement rates for each data item were calculated for individual NICUs and across the network, respectively.Results: A total of 283 cases and 24,904 data fields were re-abstracted. The agreement rates for original and re-abstracted data elements were 96.1% overall, and 97.2, 94.3, and 96.6% for critical, important, and less important data elements, respectively. Individual site variation for discrepancies ranged between 0.0 and 18.4% for all collected data elements.Conclusion: The completeness, precision, and quality of data in the CHNN database are high, providing assurance for multipurpose use, including health service evaluation, quality improvement, clinical trials, and other research. Preterm birth and low birth weight remain the single largest cause of neonatal mortality and morbidity among births worldwide . OptimizThe quality of data is essential for good epidemiological study since poor data quality may lead to unreliable conclusions . There aThe Chinese Neonatal Network (CHNN) was established in 2019, with the aim of creating a trustworthy national source of neonatal clinical information for research, benchmarking, quality improvement, and policy making. Consequently, quality assurance of the database is particularly important to make sure that the data collected are of high quality. The objective of this manuscript is to report the results of an internal audit of the CHNN database using a prospective cross-sectional study approach.CHNN was established in 2019 and comprises 58 tertiary-level neonatal intensive care units (NICU) in 25 provinces across China. All participating NICUs are grade A level III NICUs authorized by the Health Administration of China. The sites were selected to provide a large cohort representative of the different geographic regions of China. Inclusion criteria for the CHNN database were:Birth weight < 1,500 g or gestational age <32 weeks.Neonates who received the treatment for at least 24 h.Neonates who died in the NICU.Exclusion criteria were:Stillbirth.Delivery-room death.Infants transferred to non-participating hospitals within 24 h after birth.Ethics approval was received from the Ethics Committee of the Children's Hospital of Fudan University (#CHFU 2018-296) and all participating NICUs for the development, compilation, data transfer, hosting, and analysis of the CHNN dataset. Only deidentified data are transferred to the data coordinating center. All data protocols and procedures comply with national, provincial, and local regulations for protecting patients' personal privacy and confidentiality.A custom-built, stand-alone system based on MS Access was developed and used for data collection and transfer. Similar to the Canadian Neonatal Network, data are prospectively collected on maternal and neonatal demographics, antenatal and birth history, NICU admission, NICU treatment, health outcome, and hospital discharge . TrainedEach participant hospital has at least one dedicated abstractor, who is supervised by hospital site investigators and a single coordinator at the CHNN Coordinating Center, who answers questions about the process of data entry. Automated validation checks are incorporated in the data-entry software, and patient identifiers are stripped before data are submitted to the coordinating center. The central coordinator also checks the database regularly and writes the report for data quality. After submission, data are automatically checked by an error checking program and potential errors are fed back to each site for data recheck and correction . A randoData collection in the CHNN database started from Jan. 1st, 2019. The audited data was obtained from newborns who were admitted to CHNN NICUs between Jan 1, 2019 and Nov 31, 2019. During this time, there were 8,103 validated cases in the CHNN database.Three to five cases were randomly selected in each hospital based on the number of NICU admissions, i.e., three cases for smaller hospitals and five cases for larger hospitals. Within each hospital, stratified random sampling was based on duration of hospitalization in three categories\u2014short (0\u20137 days), medium (8\u201328 days), and long (>28 days) stay. The number of cases selected for audit in each category was prorated according to the distribution of cases in each category among the 58 CHNN sites. Accordingly, 53 short-stay, 108 medium-stay, and 122 long-stay cases were selected, for a total sample size of 283 cases for the data audit.We used the method of Shah et al. to dividAfter selection of variables, the coordinating center used the sampling criteria to randomly select cases from the original database for reabstraction and informed the sites accordingly. Within the following month, abstractors used a separate data reabstraction software to reenter data for cases randomly selected for data reabstraction. Abstractors were not permitted to revisit and review the original abstraction for checking of accuracy of data entry. The average time required for reabstraction of the 88 variables required for the audit was 50 min. This contrasts with the average of 60 min required for the 360 variables that are routinely abstracted for a typical patient and indicates the complexity of the variables chosen for reabstraction. Consequently, the total extra workload averaged between 2 and 3 h for each participating CHNN hospital. All CHNN hospitals participated in the data audit.Data analysis and random sampling were conducted using SAS version 9.4. Before data analysis, records of selected cases were extracted from the original CHNN database and categorized into the three abovementioned categories. The total agreement rates of each variable in these three categories were analyzed and evaluated using the following formula:For each participant site, the range of agreement rate was reported, which reflects the percentage of reabstracted record that agreed with original abstraction in each hospital of CHNN. Descriptive analysis was applied to report the median gestational age and length of stay of cases.A pilot study was conducted at the Children's Hospital of Fudan University 3 months prior to the CHNN audit exercise to test the data reabstraction software and ensure that the audit exercise could be carried out smoothly. We sampled three cases at random for the pilot study. Data analysis of the pilot study confirmed that the process could be executed efficiently.Ethics approval was obtained from the Children's Hospital of Fudan University to collect, store, and analyze the data from newborn infants admitted to all participant sites of CHNN. Ethics approvals were also obtained from all participating CHNN sites for local data collection, storage, and transfer of patient data to the CHNN Coordinating Center for analysis. All necessary health information privacy and security procedures were followed and conformed with local, provincial, and national requirements and guidelines. Prior to data transfer to the Coordinating Center, all patient identifiers were stripped to ensure privacy.During the study period, all 58 CHNN sites participated in the audit. A total of 283 cases were reabstracted. Among them, 53 cases had short duration of hospitalization (<8 days), 108 cases had medium duration of hospitalization (8\u201328 days), and 122 cases had long duration of hospitalization (>28 days). Twenty-nine cases are abstracted by different abstractors (10.25%) and 254 cases are abstracted by the same abstractors (89.75%) of the original chart abstraction. The median gestational age of audited cases was 34 weeks . The median length of stay of audited cases was 10 days , which were reflective of the overall CHNN population ranges. The agreement rate was 96.1% overall for all 88 selected data elements, 97.2% for critical data elements, 94.3% for important data elements, and 96.7% for less important data elements . When seFor critical data elements, the individual site agreement rate ranged from 83.3 to 100.0% . For indFor important data elements, the individual site agreement rate ranged from 78.6 to 100% . For indFor less-important data elements, the individual site agreement rate ranged from 82.7 to 100.0% . For indn = 98,100 variables. The agreement rate for cases with short duration of hospitalization was 96.0% , medium duration of hospitalization was 95.9% , and long duration of hospitalization was 96.1% .The overall agreement of all 360 data variables in the entire database was 96.3% for the To our knowledge, this is the first study of data quality in a neonatal database in China. This internal audit with participation of all CHNN sites showed a high rate of agreement between the reabstracted and original data collected for the CHNN database, which is the largest neonatal database of its kind in China, with participation of 58 NICUs in 25 provinces throughout China. The overall agreement rate was high at 96.1%, and the disagreement rates of 2.8% for critical variables, 5.7% for important variables, and 3.3% for less-important variables, met their prescribed targets of <5, <10, and <15%, respectively. There was low site-to-site variation in the disagreement rate, ranging from perfect agreement to 18.4% disagreement for all collected data elements. These results demonstrate that the CHNN data have high precision and reliability and is suitable for multiple uses, including research.Our results are comparable with audits performed by large neonatal networks in other countries. The Canadian Neonatal Network reported a high precision of data quality collection with small individual site variation for discrepancies 0.2\u201312.8%) through a similar internal audit process .8% throu. An asseWe found that disagreement between the reabstracted and original data were mainly due to missing maternal data and incomplete records in the patient medical charts rather than poor abstraction of data. Missing data secondary to poor patient medical chart documentation by healthcare providers is a documented phenomenon . In our The high precision and reliability of the database is due at least in part to the multiple layers of quality assurance built into the data collection system . These iThere are several limitations to this internal audit. Our audit exercise involved only a small random sample and was only conducted once per year. However, to reabstract and audit all cases in the database would be prohibitively time consuming, expensive, and impractical. Consequently, most databases only conduct small random sampling audits similar to our study , 17. An In conclusion, our audit demonstrated that data from the CHNN data collection system show high precision and reliability. With the implementation of measures designed to improve data quality based on deficiencies identified in this audit, we expect that data quality will further improve. As our network matures, periodic data audit will be essential to ensure the reliability of our database for research, and funding, manpower, and resources will be needed to support the practice of data quality improvement.The original contributions presented in the study are included in the article/Supplementary Material, further inquiries can be directed to the corresponding authors.The Ethics Committee from the Children's Hospital of Fudan University has approved CHNN to collect, store and analyse the overall data from all participant sites of CHNN. For individual sites, there is also approval from the ethics committee for local data collection and storage. Health information for each case will be well protected and maintained at local sites with a unique ID, which will not be submitted in the coordinate center. Since audit practice is the part of the current exercise of the project, specific approval will not be warranted.JS, SL, and LD: study concept and design. JS, XC, SL, and LD: acquisition, analysis, or interpretation of data. JS and XC: drafting of the manuscript. XC, YW, and XG: statistical analysis. TY and YL: administrative, technical, or material support. JS, WenZ, CC, SL, and LD: study supervision. All authors are critical revision of the manuscript for important intellectual content. All authors contributed to the article and approved the submitted version.Chairmen: Shoo K. Lee, MBBS, Mount Sinai Hospital, University of Toronto; Chao Chen, MD, Children's Hospital of Fudan University. Vice-Chairmen: Lizhong Du, MD, Children's Hospital of Zhejiang University School of Medicine; Wenhao Zhou, Children's Hospital of Fudan University. Site principle investigators of the Chinese Neonatal Network: Children's Hospital of Fudan University: Yun Cao, MD; The Third Affiliated Hospital of Zhengzhou University: Falin Xu, MD; Tianjin Central Hospital of Obstetrics and Gynecology: Xiuying Tian, MD; Guangzhou Women and Children's Medical Center: Huayan Zhang, MD; Children's Hospital of Shanxi: Yong Ji, MD; Northwest Women's and Children's Hospital: Zhankui Li, MD; Gansu Provincial Maternity and Child Care Hospital: Jingyun Shi, MD; Shengjing Hospital of China Medical University: Xindong Xue, MD; Shenzhen Maternity and Child Health Care Hospital: Chuanzhong Yang, MD; Quanzhou Women and Children's Hospital: Dongmei Chen, MD; The Affiliated Suzhou Hospital of Nanjing Medical University: Sannan Wang, MD; Guizhou Women and Children's Hospital/Guiyang Children's Hospital: Ling Liu, MD; Hunan Children's Hospital: Xirong Gao, MD; The First Bethune Hospital of Jilin University: Hui Wu, MD; Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University: Changyi Yang, MD; Nanjing Maternity and Child Health Care Hospital: Shuping Han, MD; Qingdao Women and Children's Hospital: Ruobing Shan, MD; The Affiliated Hospital of Qingdao University: Hong Jiang, MD; Children's Hospital of Shanghai: Gang Qiu, MD; Women and Children's Hospital of Guangxi Zhuang Autonomous Region: Qiufen Wei, MD; Children's Hospital of Nanjing Medical University: Rui Cheng, MD; Henan Children's Hospital: Wenqing Kang, MD; The First Affiliated Hospital of Xinjiang Medical University: Mingxia Li, MD; Foshan Women and Children's Hospital: Yiheng Dai, MD; The First Affiliated Hospital of Anhui Medical University: Lili Wang, MD; Shanghai First Maternity and Infant Hospital: Jiangqin Liu MD; Yuying Children's Hospital Affiliated to Wenzhou Medical University: Zhenlang Lin, MD; Children's Hospital of Chongqing Medical University: Yuan Shi, MD; The First Affiliated Hospital of Zhengzhou University: Xiuyong Cheng, MD; The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China: Jiahua Pan, MD; Shaanxi Provincial People's Hospital: Qin Zhang, MD; Children's Hospital of Soochow University: Xing Feng, MD; Wuxi Maternity and Child Healthcare Hospital: Qin Zhou, MD; People's Hospital of Xinjiang Uygur Autonomous Region: Long Li, MD; The Second Xiangya Hospital of Central South University: Pingyang Chen, MD; Qilu Children's Hospital of Shandong University: Xiaoying Li, MD; Hainan Women and Children's Hospital: Ling Yang, MD; Xiamen Children's Hospital: Deyi Zhuang, MD; Xinhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine: Yongjun Zhang, MD; Shanghai Children's Medical Center, School of Medicine, Shanghai Jiao Tong University: Jianhua Sun, MD; Shenzhen Children's Hospital: Jinxing Feng, MD; Children's Hospital Affiliated to Capital Institute of Pediatrics: Li Li, MD; Women and Children's Hospital, School of Medicine, Xiamen university: Xinzhu Lin, MD; General Hospital of Ningxia Medical University: Yinping Qiu, MD; First Affiliated Hospital of Kunming Medical University: Kun Liang, MD; Hebei Provincial Children's Hospital: Li Ma, MD; Jiangxi Provincial Children's Hospital: Liping Chen, MD; Fuzhou Children's Hospital of Fujian Province: Liyan Zhang, MD; First Affiliated Hospital of Xian Jiao Tong University: Hongxia Song, MD; Dehong people's Hospital of Yunnan Province: Zhaoqing Yin, MD; Beijing Children's Hospital, Capital Medical University: Mingyan Hei, MD; Zhuhai Center for Maternal and Child Health Care: Huiwen Huang, MD; Guangdong Women and Children's Hospital: Jie Yang, MD; Dalian Municipal Women and Children's Medical Center: Dong Li, MD; Peking Union Medical College Hospital: Guofang Ding, MD; Obstetrics & Gynecology Hospital of Fudan University: Jimei Wang, MD; Shenzhen Hospital of Hongkong University: Qianshen Zhang, MD; Children's Hospital of Zhejiang University School of Medicine: Xiaolu Ma, MD.This work was funded by the Canadian Institutes of Health Research [CTP87518 to SL].The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Calcium supplementation can prevent gestational hypertension and pre-eclampsia. However, besides the non-consensus of existing studies, there is a lack of evidence regarding the optimal dosing of calcium.Eight electronic databases, namely, the Cochrane Library, PUBMED, Web of Science, EMBASE, WANGFANG, VIP, CBM, and CNKI, were searched. The studies were retrieved from inception to July 13, 2021. Two researchers independently screened the literature, extracted data, and evaluated the methodological quality based on the inclusion criteria. In particular, the calcium supplementation doses were divided into three groups, namely, the high-dose (\u22651.5 g), medium-dose (1.0\u20131.49 g), and the low-dose group (<1.0 g). The participants were also divided into high-risk and low-risk groups, according to the risk of developing gestational hypertension and pre-eclampsia.A total of 48 studies were incorporated into the final analyses. All doses of calcium supplementation reduced the incidence of gestational hypertension in the low-risk population , whereas the medium-dose (three studies) reduced the incidence of gestational hypertension in high-risk groups. Moreover, a medium dose of calcium supplementation had the maximum effect in reducing gestational hypertension in low-risk and high-risk populations. The medium (three studies) and high doses (13 studies) of calcium supplementation reduced the incidence of pre-eclampsia in the low-risk groups. However, a medium-dose calcium supplementation maximally prevented pre-eclampsia in the low-risk population. The authenticity and reliability of the results were reduced due to the limitations of contemporary studies in terms of experimental design, result measurement, statistics, and evidence quality. Therefore, high-quality studies with larger sample size are required to evaluate further the effect of calcium supplementation in preventing gestational hypertension and pre-eclampsia. Hypertensive disorders of pregnancy (HDP) are one of the three leading causes of global maternal morbidity and mortality. The incidence of HDP increased from 16.30 to 18.08 million globally, with a total increase of 10.92% from 1990 to 2019 . In addiMoreover, as the condition progresses, pre-eclampsia can quickly develop into eclampsia, which leads to life-threatening convulsions and coma. Notably, gestational hypertension and pre-eclampsia are known to threaten the maternal and newborn's health significantly, which eventually causes considerable suffering and substantial economic burden on patients . MeanwhiNumerous epidemiological investigations have revealed that insufficient calcium intake leads to a significant increase in morbidity and mortality during gestational hypertension and pre-eclampsia , 11. In Results of multiple meta-analyses showed that , 18, 20 The following inclusion and exclusion criteria were used based on the definition of participants, interventions, comparisons, outcomes, and study (PICOS) in the Cochrane Systematic Review Manual.Pregnant women without gestational hypertension and pre-eclampsia were included, with no limitation to whether people were at a high risk of HDP. Based on the risk factors of HDP, pregnant women were divided into two subgroups: High-risk group: Pregnant women with calcium deficiency, a history of gestational hypertension or pre-eclampsia, a positive roll-over test, a positive angiotensin-sensitivity test, or a high-risk factor defined in the original study . Low-risThe test group was treated with calcium supplementation. The dose and the dose setting were described in the published literature . FurtherThe control group was treated with a placebo or blank control.The incidence of gestational hypertension and pre-eclampsia.Randomized controlled trials.Studies of calcium combined with other supplements were excluded ; repeatedly published studies; non-Chinese and English literature; and no access of full text and/or studies with incomplete data.We searched scientific databases such as the PubMed, Ovid-Embase, The Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), Chinese Scientific Journal Database (CSJD-VIP), Wanfang Database, and the China Biomedical Literature Database (CBM). The relevant literature was retrieved from inception to July 13, 2021. The search terms were AND . Two trained researchers selected the articles and stringently extracted the data based on the inclusion/exclusion criteria, and the selections were cross-checked. In the case of disagreement, a third researcher (Rongxia He) settled the conflict with a common consensus. Data were extracted according to the pre-established full-text data extraction checklist, including (1) Basic characteristics of studies such as authors, publication years, country, source and age of patients, start time of calcium supplementation, disease risk, dietary calcium intakes, and the number lost for follow-up. (2) Key elements of bias risk assessment. (3) Outcome measures: The incidence of gestational hypertension and pre-eclampsia.Using the RCT bias risk assessment tool recommended by the Cochrane Handbook for Systematic Reviews of Interventions 5.1.0 , two traP = 0.1.). Also, I2 was used to judge the degree of heterogeneity. The fixed-effect model was used for meta-analysis if the research results were not statistically different. Conversely, if there were statistical heterogeneity, the source of heterogeneity was further analyzed, and the random-effects model was used for meta-analysis after the exclusion of evident clinical heterogeneity. The significance level for tests was set at 0.05.STATA 16 software was used for statistical analysis. Risk ratio (RR) was used to study the effect analysis statistic for dichotomous variables and the 95% CI as the effect amount. Heterogeneity of results between studies was assessed by a Chi-square test . If PSRF was close to 1, the convergence of this study was good, and the conclusion of the meta-analysis was reliable. Also, the network group commands were used for data pre-processing based on STATA 16 software to compare the outcome indicators of the network relationship between the intervention measures.GeMTC-0.14.3 software based on the Bayesian model was used for statistical analysis. The software used Markov chain\u2013Monte Carlo (MCMC) to prioritize and evaluate the data to achieve reticular meta-analysis. The conditions set by the network meta-analysis were as follows: Number of chains: 4, Tuning iterations: 20,000, Simulation iterations: 50,000, Thinning interval: 10. Inference samples: 10,000, Variance scaling factor: 2.5. The deviation information criterion value of the random effect model and fixed effect model were compared to analyze the fitting degree of the model. The risk ratio (RR) was selected as statistics for effect, and a 95% CI was used. The network meta-analysis used the concordance model, which was statistically significant. The node analysis model was used for the inconsistency test; if A total of 11,981 related articles including 4,965 Chinese and 7,016 English records were obtained. After excluding the literature based on the exclusion criteria, eventually 48 studies were included. The studied 48 studies include 24 Chinese articles \u201346 and 2A total of 48 randomized controlled trials were included for analysis. The number of patients ranged from 30 to 9,178Among the included 48 studies, only 17 studies , 64\u201366 rA total of 37 studies , 61\u201365 rn = 3 studies, RR = 0.27 ; medium dose: n = 11 studies, RR = 0.35 ; high dose: n = 28 studies, RR = 0.48 ] ] .The network meta-analysis revealed that the different doses of calcium supplementation reduced the gestational hypertension incidence; however, no significant difference was observed in various doses ] . As only, 0.84)] .The network meta-analysis revealed that the medium doses of calcium supplementation reduced the incidence of gestational hypertension. Conversely, both low and high doses could not reduce the incidence of gestational hypertension. Of note, no significant difference was observed among different doses in reducing the incidence of gestational hypertension . EvidencA total of 20 studies , 58\u201366 rn = 4 studies, RR = 0.49 ; medium dose: n = 3 studies, RR = 0.32 ; high dose: n = 13 studies, RR = 0.67 ] ] .The network meta-analysis showed that medium and high doses of calcium supplementation reduced the incidence of pre-eclampsia. Conversely, the low dose could not reduce the incidence of pre-eclampsia. The effect of medium and high doses of calcium supplements was significantly better than the low doses The int. Evidencn = 3 studies, RR = 0.21 ] ] .Compared with the previously published meta-analysis, our study compiled all the relevant literature of the eight Chinese and English databases. According to the inclusion and exclusion criteria formulated in advance, the literature was strictly screened. Finally, data of 48 comprehensive studies were included for the final merger analysis. Meanwhile, we grouped these populations into two risk levels: low risk and high risk. For subgroup analysis, the population was further grouped into three levels of a high, medium, and low, based on the calcium doses. Due to the lack of evidence for direct comparison between different calcium doses, we indirectly compared the effectiveness of different calcium doses in reducing gestational hypertension and pre-eclampsia by network meta-analysis to find evidence of the optimal calcium dose. Overall, our meta-analysis results were more stable and reliable.For gestational hypertension, the results of the traditional meta-analysis were consistent with the network meta-analysis for the low-risk population, indicating that different doses of calcium supplementation reduced the incidence of gestational hypertension. In the low-risk groups, the low dose of calcium supplementation had met the calcium demand of pregnant women, and excessive calcium doses would inhibit iron absorption, thus affecting the overall development of the disease. Therefore, excessive doses of calcium supplementation did not show significant extended benefits , 68. TheFor pre-eclampsia, the results of our traditional meta-analysis were consistent with the previous reports by Sun et al., where the different doses of calcium supplementation could reduce the incidence of pre-eclampsia in low-risk groups . AlthougIn summary, both medium and high doses of calcium supplements prevented pre-eclampsia in low-risk populations; however, given the economic cost and drug side effects, a medium dose was the most effective in preventing pre-eclampsia , 71. On Our study found that calcium supplementation for gestational hypertension and pre-eclampsia was not substantial based on a rigorous systematic review of current literature. Besides, the reliability of the results was reduced, which eventually reduced the authenticity of meta-analysis results. Probable reasons for this observation are discussed as follows:Heterogeneity of the included studies: In high-risk groups, the baseline characteristics of maternal age (including 18\u201330 years old), initiation of calcium supplementation (including 14\u201330 weeks of pregnancy), and high-risk factors were significantly different. These factors eventually caused more significant heterogeneity among the studies and reduced the reliability of meta-analysis results.Inadequate rigorous and scientific design of studies: Among the 48 included studies, the random grouping method of 64.58% (31/48) of the studies was unclear, whereas 75% (36/48) of the studies did not report the implementation of covert grouping. This discrepancy led to the high likelihood of selective bias. Thus, a stricter grouping method should be considered for future research to reduce selection bias.Inclusion studies lack important quality control measures to reduce measurement and implementation bias: A total of 62.5% (30/48) of the studies did not report blinding methods for patients, researchers, and outcome evaluators. For the determination of the outcome index, an effective scientific blinding method could avoid the influence of measurement bias on the measurement results, the qualification of the surveyors, the consistency of the surveyor's cognition of different outcome indicators, the accuracy, and scientific nature of the effectiveness criteria could affect the determination of the results in varying degrees. However, none of the 48 studies reported the qualifications of the surveyors or the standards and specific measurement processes used in the study. Therefore, future research should apply the blinding method for the experimental designs and report specific and comprehensive experimental details. This practice would improve the reproducibility and reliability of study results.Unbiased report of study conditions and data: Although the included 48 studies reported all the predetermined results in their publications, only 18.75% of the studies reported the research protocols. Due to this discrepancy, it was impractical to finally judge whether to report all their results as per the plan and without bias. Selective reporting of the research results could lead to publication bias, which eventually affects the reliability of systematic review conclusions and even prompts an opposite conclusion. In addition, although most studies have reported calcium supplementation, the specific components of calcium supplements were not reported. The variation in the formulation may result in a deviation between the reported and the real dose of calcium supplementation for pregnant women, thus affecting the reliability of the conclusions of our study and pre-eclampsia (low-risk population). Unfortunately, only a few studies are reported on preventing gestational hypertension and pre-eclampsia for high-risk groups. Therefore, more high-quality clinical studies are required to further explore the specific role of calcium. Moreover, future research should be diligently carried out in study design, implementation, measurement, and evaluation of results and research protocols, to improve the quality of studies.Advantages of this study: (1) The subgroup analysis of the included population was carried out for the different risk groups and calcium supplementation doses, which improved the applicability of the meta-analysis results. (2) An indirect comparison of the effects of different calcium doses yielded evidence for optimal calcium doses. (3) As more studies were included, the conclusion was more comprehensive and reliable.Limitations of this study: (1) Most studies did not report daily dietary calcium intake in pregnant women. Thus, our study could not consider the effect of dietary calcium intake on calcium supplementation during pregnancy. Moreover, the people with adequate dietary calcium intake might have a smaller response to the effect of calcium supplementation , 69. (2)All doses of calcium could reduce the incidence of gestational hypertension in a low-risk population, whereas medium dose could reduce the incidence of gestational hypertension in high-risk groups. Moreover, in the comprehensive analysis of the included studies, the medium dose was most effective (low-risk and high-risk populations). Both medium and high doses of calcium could reduce the incidence of pre-eclampsia in low-risk groups. However, a medium dose was found to have maximum effect. The bias was caused by the limitations of included studies regarding outcome measurement and reporting, whereas the fewer data on some indicators reduced the authenticity and reliability. Therefore, future trials need to design research programs more scientifically and rationally by including large cohorts. This intervention would reduce the risk of bias, improve the quality of evidence, and further evaluate the efficacy of calcium supplementation.The original contributions presented in the study are included in the article/RH contributed to the design, guidance, and modification of the project. DC and HW completed the implementation and writing of the paper. XX, LZ, AY, and SL completed the collection and collation of the data. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Correction to: Implement Sci 16, 37 (2021)https://doi.org/10.1186/s13012-021-01106-2Following publication of the original article , the autThis publication was supported in part by the Centers for Disease Control and Prevention of the U.S. Department of Health and Human Services (HHS) as part of a financial assistance award. The contents are those of the author(s) and do not necessarily represent the official views of, nor an endorsement, by CDC/HHS, or the U.S. Government."} +{"text": "Diffuse large B-cell lymphoma (DLBCL) is the most common histopathological type of non-Hodgkin\u2019s lymphoma, which may arise from various extranodal sites. Little is known about the clinical characteristics and survival outcomes of primary DLBCL of the urinary tract (UT). Thus, we conducted this study to explore the independent prognostic factors of patients with UT-DLBCL using the Surveillance, Epidemiology, and End Results (SEER) database.We searched the Surveillance, Epidemiology, and End Results (SEER) database for the data of patients diagnosed with UT-DLBCL between 1975 and 2016. Data, including demographic tumour stage and therapeutic strategies, such as surgical resection, radiation therapy, and chemotherapy, were collected. The impact of these factors on survival outcomes, including overall survival (OS) and disease-specific survival (DSS), was analysed using Kaplan\u2013Meier curves.Four-hundred and eighty-nine patients who met the inclusion criteria were enrolled in the data analysis. The median age was 69 years old. Most cases of UT-DLBCL (72.39%) originated from the kidney, followed by the urinary bladder (24.95%). Both surgical resection and chemotherapy can significantly improve OS and DSS. Patients older than 75 years had the worst survival outcomes. Stage IV DLBCL may be a poor prognostic factor.To the best of our knowledge, this is the largest population-based study of UT-DLBCL. Advanced age, male gender, lack of surgical resection or chemotherapy, and stage IV DLBCL were poor prognostic factors. The most commonly diagnosed non-Hodgkin\u2019s lymphoma (NHL) subtype is diffuse large B-cell lymphoma (DLBCL), which accounts for approximately 30% of NHL cases . LymphomThe use of recently established first-line therapy, including rituximab combined with cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP), has improved the survival outcomes of patients with DLBCL . HoweverThis study explored the prognostic factors of patients with UT-DLBCL, based on data from the SEER database, to provide findings that will facilitate better clinic evaluations and more effective management of DLBCL.We utilized the SEER database to enrol eligible patients between 1975 and 2016, collected from 18 state registries in the USA. The patients diagnosed with lymphoma with the third edition of the International Classification of Diseases for Oncology (ICD-O-3) histology code of 9680 in the database were included. Patients with DLBCL originating from sites other than the UT were excluded. In addition, all cases diagnosed before 1983 were excluded for the lack of the Ann-Arbor stage information. The patients without active follow-up and unknown Ann-Arbor stage, and those without histopathological confirmation of DLBCL, were also excluded from this study. Eligible patients were further identified based on the primary site, such as the kidney, ureter, urinary bladder, and urethra.All the demographic, clinical, pathological, and survival data of the patients were extracted from the SEER database. The patients were divided into two groups based on the primary site: upper UT (UUT) group, with sites that include kidney, renal pelvis, and ureter; and lower UT (LUT) group, with sites that include urinary bladder and urethra. The treatments for patients with UT-DLBCL were also exported from the database, including radiotherapy, cancer-directed surgical resection, and chemotherapy. The ages of diagnosis were grouped into the following: 0\u201360, 60\u201375, and older than 75 years. The races were classified into white, black, and other. Importantly, the overall survival (OS) and disease-specific survival (DSS) were both analysed in this study.The SEER database records survival as the number of months elapsed from the time of diagnosis of DLBCL to the day of death or the latest contact. For the estimation of disease-specific survival (DSS), only death attributable to the DLBCL was considered an exact event. To prevent confounding, we excluded patients diagnosed with more than one primary malignancy from the survival analysis.All data used in this study were from the public database, thus, ethical approval was required. We maintained the confidentiality of the data and reported the study results according to the SEER Research Data Use Agreement.Continuous variables with normal distribution are presented as mean and standard deviation (SD); otherwise, median and inter-quartile range are considered. The differences in continuous variables were evaluated using the 2-sided analysis of variance (ANOVA) or the Mann-Whitney U-test according to their distribution. The categorical variables are presented as counts and percentages. Chi-squares or Fischer\u2019s exact tests were applied to evaluate their differences. The survival curves were estimated using the Kaplan\u2013Meier method and compared using the log-rank test. Cox proportional hazard models were used to identify the independent predictors of disease-specific mortality. Patients with missing data for one or more of the variables were excluded from the multivariable regression analysis. To allow adequate power for the Kaplan\u2013Meier curves and the Cox model, the kidney cases were grouped with renal pelvis and ureter cases, whereas the urethral cases were grouped with the bladder cases. The statistical analysis was performed using IBM SPSS Statistics 24 software , and a p-value of less than 0.05 was considered statistically significant.Finally, we enrolled 489 eligible patients diagnosed with UT-DLBCL between 1975 and 2016 from the SEER database. The demographic and clinical characteristics of the patients are summarized in Surprisingly, cancer-directed surgery was performed in about half of the cases (56.44%). Forty-one patients (7.77%) underwent radiation therapy. The majority of these cases (72.80%) underwent chemotherapy. The most common Ann-Arbor stage at presentation was stage I (34.15%), followed by stage IV (34.35%).Patients aged \u2265 75 years had the worst survival outcomes, with OS of only 8 months and DSS of 12 months . The same outcome was found for DSS in the two groups .Publicly available datasets were analyzed in this study. This data can be found here: Surveillance, Epidemiology, and End Results (SEER) database and the National Natural Science Fund of China (81771569).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "There was an error in the spelling of an author\u2019s name; Juan M. Abofalia should be Juan M. Abolafia.The was also an error in one of the grant numbers in the funding statement for this article. The funding statement should read as follows:\u201cThis work was supported by EU H2020 Research and Innovation Programme under Grant Agreement No. 945539 (HBP SGA3), BFU2017-85048-R Spanish Ministry of Science to MVSV and by the project Async-Prop, Bournemouth University-IDIBAPS, RED11549, Partnering Project of the HBP SP3 (EU H2020 Research and Innovation Programme) to EB-B. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u201d"} +{"text": "Enterprise Risk Management (ERM) in healthcare is a method used to identify, assess and reduce risk to patients and the hospital organization. The objective of this study is to identify clinical and organizational challenges and risks in healthcare management caused by COVID-19, and its impact on patients and healthcare workers, in a low-resource obstetric setting.From a census of patients from 1 April 2020 to 30 July 2020, four cases of COVID-19 in pregnancy representing different severity levels were selected. A patient tracer activity was done for each patient, documenting events that the patient and healthcare team experienced from admission to discharge. A case series on these patients was written. A focus group consisting of an OB-GYN resident, OB-GYN consultant, OB-GYN nurse, OB-GYN infectious disease consultant, and internal medicine resident and consultant, was formed. Each case was presented to the focus group to establish the context of risk assessment. Risks were identified using the framework of Enterprise Risk Management. Each risk was classified according to their risk domain and severity. Root cause analysis via the fishbone method was used to identify the causes of the risks.Operational risks identified were delayed swab results, false negative swab results, and delayed patient transport. Clinical/Patient risks identified were COVID-19 exposure of healthcare workers and other non-COVID patients, inadvertent community exposure, risk for severe clinical manifestations of COVID-19, and lack of specific treatment for COVID-19. Risk to human capital identified were COVID-19 infection of hospital staff and decreased quantity of workforce due to quarantine. Most risks were assessed to be moderate risk or high risk in terms of severity. Root cause analysis showed that common causes of risks were due to exposure to asymptomatic patients and delayed and false-negative swab results.The results of this study may be used towards the final steps of ERM: risk evaluation, treatment and management, in a low resource setting.Judith P. Peralta, Doctor of Medicine, Fellow of Philippine Obstetrical and Gynecological Society, Fellow of Philippine Infectious Disease Society for Obstetrics and Gynecology, Research Fellow - Harvard Medical School Department of Obstetrics and Gynecology and Reproductive Medicine (2014-2015), Member - International Society of Infectious Diseases , Pfizer"} +{"text": "To verify, through a systematic review, the accuracy of nutritional assessment in children and adolescents using the length/height-for-age and BMI-for-age growth charts of the Centers for Disease Control and Prevention (CDC) (2000), the World Health Organization (WHO) (2006/2007) and the International Obesity Task Force (IOTF) (2012).Medical Literature Analysis and Retrieval System Online (MEDLINE), through PubMed, National Library of Medicine and The National Institutes of Health (NIH), Scientific Electronic Library Online (SciELO) and Virtual Health Library (VHL). The following descriptors were used for the search: \u201cChild\u201d, \u201cAdolescent\u201d, \u201cNutritional Assessment\u201d, \u201cGrowth Chart\u201d, \u201cEthnic Groups\u201d, \u201cStature by age\u201d, \u201cBody Mass Index\u201d, \u201cComparison\u201d, \u201cCDC\u201d, \u201cWHO\u201d, and \u201cIOTF\u201d. The selected articles were assessed for quality through the Quality Assessment Tool for Observational Cohort and Cross-Sectional Studies of the NIH.We selected articles from the databases Thirty-three studies published between 2007 and 2020 were selected and, of these, 20 presented good quality, 12 presented fair quality and one presented poor quality. For children under five years old, the WHO length/height-for-age growth charts were shown appropriate for children from Argentina, South Africa, Brazil, Gabon, Qatar, Pakistan and the United States. For those five years old and older, the WHO BMI-for-age growth charts were accurate for the Brazilian and Canadian populations, while the IOTF growth charts were accurate for the European populations.There are difficulties in obtaining international growth charts for children from 5 years old and older that go along with a long period of growth, and which include genetic, cultural and socioeconomic differences of multiethnic populations who have already overcome the secular trend in height. These growth charts were created based on longitudinal and/or cross-sectional studies with samples of children and adolescents considered a reference or standard. They express distributions in percentiles or Z scores and are considered quite sensitive for the assessment of nutritional status, enabling interventions and the prevention of health problems.For decades, the precision in assessing the growth of children and adolescents has been the object of study by several researchers, who use anthropometry and growth charts to monitor the evolution of growth changes and to assess the nutritional status of children under 20 years of age. Among these, the growth charts by the Centers for Disease Control and Prevention (CDC) (2000), the World Health Organization (WHO) (2006/2007) and the International Obesity Task Force (IOTF) (2012) stand out.Different growth charts have been proposed by some institutions and organizations over the years for use in the world population, through studies with national or international samples and with different inclusion criteria. They are expressed in percentiles and are specific by sex and age group. For children under three years of age, there are growth charts of length-for-age, weight-for-age and head circumference-for-age. For children under five, there is a weight-for-height growth chart, and for children and adolescents aged between two and 20 years, there are growth charts representing stature-for-age, weight-for-age and body mass index (BMI) for age.The CDC growth charts were drawn up in the 2000s based on five national surveys conducted in the United States. This work was conducted in six countries: Brazil (Pelotas), United States (Davis), Ghana (Accra), Norway (Oslo), India (New Delhi) and Oman (Muscat) with children considered standard, that is, who lived in socio-environmental and economic conditions ideal for an adequate development. These growth charts were constructed based on longitudinal (from birth to two years old) and cross-sectional samples with children aged 18 to 71 months. For children aged five years or more and adolescents aged up to 20 years, the construction of the growth charts was based on the cross-sectional study of the National Center for Health Statistics (NCHS/1977), whose only study population was from the United States. For the construction of these growth charts, the WHO specialists committee remodeled the 1997 NCHS data, keeping only non-obese children and adolescents who had reached expected heights for their age and adding growth patterns data for under-fives aged 18 to 71 months. The addition of these data smoothed the growth charts, creating a smooth transition at five years of age and at the end of adolescence, with adjustment to the overweight and obesity cutoff points recommended for adults.The WHO growth charts for children under the age of five were developed in 2006 based on the Multicenter Growth Reference Study, whose goal was to describe the growth of healthy children. For children under five, there are head circumference-for-age and weight-for-height growth charts.2,6 For children under 10 years old there is the weight-for-age growth chart and, for children and adolescents under 20 years there are the length/height-for-age and BMI-for-age growth charts.2,3,6,7,10The WHO growth charts are expressed in percentiles or Z-scores and are specific for sex and age group. for 18 years, suggesting classifications distributed by age and sex, as well as overweight and obesity classifications. In 2012, after studies showed divergences in the WHO growth reference (2006/2007) in some populations, the IOTF released an update of its cutoff points using international samples and proposed these for the BMI, which resulted in six different classifications similar to WHO\u2019s, ranging from severe thinness to morbid obesity.Still in the 2000s, the IOTF developed the BMI-for-age growth charts for children and adolescents aged between two and 20 years, with BMI values of 25 and 30 kg/m These indicators have the following objectives, respectively: a) to show the linear trajectory of growth, being fundamental in the detection of stunting; b) to detect underweight or overweight. The cutoff points of the CDC (2000), the WHO (2006/2007) and the IOTF (2012), in percentiles, for the length/height -for-age and BMI-for-age indicators are shown in The two main anthropometric indicators used in the assessment of children and adolescents are length/height-for-age and BMI-for-age. However, some studies have shown divergent comparisons between the national growth charts and the WHO growth charts.4 Examples are places like the United Kingdom, Poland, Norway, Germany, Hong Kong, Iran, United Arab Emirates and South Africa. For this reason, the United Kingdom created growth charts for certain ages based on the joining of the WHO growth references with local data, while countries such as China, Bolivia, Denmark, Norway and Belgium, have not used the WHO growth charts widely due to divergences in growth parameters of their populations when compared to the reference growth charts.4,15WHO recommends its own growth charts (2006/2007) for international use, and they have been adopted in health and nutrition programs in more than 140 countries, including Brazil. These differences generate effects on the accuracy of nutritional classification and, by extension, make diagnosis and comparison of prevalence difficult.The methodological differences in establishing cutoff points between the CDC, WHO and IOTF references involve population composition and modeling of descriptive parameters of the anthropometric index and cutoff points. Furthermore, variation in body composition between children and adolescents of different ethnicities has been an obstacle to the determination of an international standard for classification of nutritional status. Thus, the objective of this study was to verify, through a systematic review, the accuracy of nutritional assessment in children and adolescents based on the growth charts recommended for international use of length/height-for-age and BMI-for-age by the CDC (2000), WHO (2006/2007) and IOTF (2012).Some authors justify that these growth charts should be based on local populations, since there are genetic, cultural and socioeconomic differences that impact the processes of physical growth and biological maturation, which result in different growth profiles and BMI. This project was registered in the International Prospective Register of Systematic Reviews (PROSPERO) under protocol CRD42020215498, and the data and outlines of this review can be accessed at www.crd.york.ac.uk/PROSPERO/display_record.asp?ID=CRD42020215498.This study is characterized as a systematic literature review, designed in accordance with the recommendations proposed by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA).The Participants, Intervention, Comparison, Outcome, Study Design (PICOS) strategy was applied for the selection of studies. We considered the studies that evaluate: P (children and adolescents), I , C , O , S .Two independent researchers consulted articles published in Portuguese, Spanish and English between 2000 and 2020 in the Electronic Medical Literature Analysis and Retrieval System Online (MEDLINE) databases, via PubMed, National Library of Medicine and The National Institutes of Health, Scientific Electronic Library Online (SciELO) and Virtual Health Library (VHL). In the search strategy, the terms of the Medical Subject Headings (MeSH) and the Health Sciences Descriptors (DeCS) used were: \u201cchild\u2019\u2019, \u201cadolescent\u201d, \u201cnutritional assessment\u201d, \u201cgrowth charts\u201d, \u201cethnic groups\u201d, \u201cstature by age\u201d, \u201cbody mass index\u201d, \u201ccomparison\u201d, \u201cCDC\u201d, \u201cWHO\u201d and \u201cIOTF\u201d .Studies were considered eligible for inclusion when they met the following criteria: a) evaluated the CDC (2000) and/or WHO (2006/2007) length/height-for-age growth charts in children and/or adolescents; and/or b) evaluated the BMI-for-age growth charts of the CDC (2000) and/or the WHO (2006/2007) and/or the IOTF (2012) in children and/or adolescents. The selection of evidence was restricted to original articles, excluding review studies, experimental studies with animals, case reports, duplicate studies and studies published in languages other than those mentioned above.The selection was first conducted by means of titles, then abstracts and, finally, full reading. The three steps were performed by two evaluators, who decided on inclusion in each step based on the eligibility criteria. Each evaluator independently decided for \u201cinclusion\u201d or \u201cexclusion\u201d and any divergent results were analyzed by a third evaluator. Eligible studies had their data extracted independently by two authors, who organized them in instruments built for this purpose, following methodological recommendations and contemplating the following items: identification of original article, study design, study population, sample size and main results related to the evaluated indicators/references. This instrument suggests the classification of quality as good, fair, and poor based on the analysis of 14 items. To assess the studies included in this review, eight items of this scale were used, referring to study objectives, study population, selection criteria, statistical power of the sample, intervention/exposure measures, loss to follow-up and outcome.The quality of the articles was assessed by adapting the Quality Assessment Tool for Observational Cohort and Cross-Sectional Studies, appropriate for observational studies, by the NIH.In October 2020, 184 articles published between 2000 and 2020 were identified in the databases. After selection by title, 91 studies were excluded, with 93 articles remaining for the abstract analysis. Fifty-five articles were selected for full reading, of which 33, published between 2007 and 2020, were included in the synthesis of evidence for this review.Studies from several countries were identified addressing the application of international growth charts in their populations. To present the results of this review, we grouped the synthesis of findings of the 33 evidences of studies that evaluated growth charts for children under five years old , for chi However, for the population of Sri Lanka, researchers state the need for further studies, since children in that country presented lower height when compared to the WHO\u2019s standards population.For children under five years old, studies show that the WHO length/height-for-age charts performed better in detecting stunting when compared to CDC growth charts.19-26,28,29 For this reason, the authors recommended the WHO growth charts for populations in Argentina, South Africa, Brazil, Gabon, Qatar, Pakistan and the United States. Some authors argue that the WHO has constructed growth charts for children under five years old based on multiethnic children who had adequate health and nutrition conditions and who received exclusive breastfeeding with until at least three or four months of age, and complementary feeding based on legumes, meat, eggs, fruits and vegetables, with partial breastfeeding, until the 12th month of life or more, which allows these growth charts to be applied internationally and to early diagnose stunting, overweight and obesity, being more accurate than those of the CDC.Regarding the BMI-for-age charts in children under five years old, the WHO diagnosed more children with underweight than the CDC for the US population and more overweight and obese children in South Africa and Brazil, which indicates that they are more appropriate for these populations. Immigrants from South Asia living in the Netherlands had lower height-for-age values than WHO\u2019s standard population, while the populations of Australia, Slovakia and Germany presented higher height values, which indicates that this international reference would not adequately detect stunting for children and adolescents (\u22655 years) of these populations. Similar results were found by Bonthuis et al. in a study that evaluated 18 national height-for-age charts from 28 European countries and compared them with those of the CDC, WHO, and Euro-Growth. The authors report that these national European growth charts showed a positive secular trend in height, which has been observed since 1850, and that this secular trend has slowed down or even reached a plateau since the 1980s/1990s in many northern European countries, as well as in Italy and the United States. In addition, the authors reinforce that, although these divergences are associated with genetic and geographic factors, they are strongly affected by the secular trend in height, and that height growth charts constructed with data collected before 1990, including those from the CDC and WHO/2007, produced mean heights generally lower than those in growth charts developed more recently. Therefore, they advocate the use of specific growth charts for the European population based on recent national data.For children and adolescents aged five years or more, studies show that the WHO height-for-age charts have similar values only for the Brazilian population. From another perspective, for the Asian populations of China, Saudi Arabia and Iran, there is great variation between the WHO and national growth charts. When compared to the WHO standard/reference populations, Chinese boys present higher weight values and Chinese girls lower weight values, with significant variations in some age groups, while children and adolescents from Saudi Arabia present higher percentile values.Regarding the WHO/2007 BMI-for-age charts for the Brazilian population, they were adequate for diagnosing overweight and obesity, being similar to the Brazilian national growth charts (Conde & Monteiro), and showing substantial agreement with those of the IOTF.. Regarding the CDC BMI-for-age growth charts, their values were similar to those of WHO/2007 for the Canadian population and similar to those of the IOTF for the Portuguese population. On the other hand, they diagnosed more overweight in South Africa and overestimated the diagnoses of overweight, obesity and underweight in Saudi Arabia and underweight in Brazil, while underestimating the diagnoses of overweight in Brazil and obesity in Iran. ..Regarding the IOTF growth charts, for the European populations of Slovakia, Italy, Poland and Portugal, they showed the best performance for screening overweight and obesity, while for the population of South Africa they had the best screening for underweight. In their study. although there was a substantial agreement between the CDC, IOTF and WHO growth charts for the classification of nutritional status of adolescents, those of the IOTF classified more overweight compared to other international references, while the WHO classified more adolescents as overweight and less as obese compared to CDC. From another perspective, in a study conducted in El Salvador by P\u00e9rez et al. with children aged six to nine years, despite the strong agreement between the WHO and IOTF growth charts, the WHO growth reference classifies more overweight and obese children than the IOTF.These variations in nutritional diagnosis caused by different BMI-for-age growth charts are in line with the findings of a study conducted by Li et al. with the population of the United States. However, other authors argue that the use of a single population in the modeling of growth charts makes them not suitable for international use and, therefore, they suggest the application of IOTF growth charts, as they were developed by combining the most recent BMI data of children aged 2-18 years from six nationally representative surveys from 1963 to 1993.Overall, the CDC BMI-for-age growth charts underperformed for screenings of nutritional diagnoses than the growth charts by WHO and IOTF. However, there is still controversy as to which growth reference would be more appropriate for international use, especially for children from five years old and over. Some authors argue that the WHO/2007 growth reference consists of a non-obese sample of children in the United States aged 1-24 years with data collected from 1963 to 1974, being a reference population that represents a healthier group and, therefore, more sensitive to diagnoses of overweight. Some studies included in this review had limitations such as the absence of sample size and statistical power in cross-sectional studies and the loss to follow-up in cohort studies, although such restrictions have not influenced in the results of this review, given the good methodological quality achieved.Regarding the quality of the selected studies, it was considered excellent, with most studies classified as having good methodological quality, as shown in This systematic review allowed, for the first time, the identification and assessment of accuracy of the length/height-for-age international growth charts by CDC and WHO, and BMI-for-age growth charts by CDC, WHO and IOTF in 20 countries from five different continents. Per this investigation, for children under five years old, the WHO length/height-for-age growth charts were proven more accurate than those of the CDC and, therefore, more appropriate for use in the populations of Argentina, South Africa, Brazil, Gabon, Qatar, Pakistan and the United States; the WHO BMI-for-age growth charts also showed better screenings of nutritional status when compared to the CDC, being recommended for the populations of the United States, South Africa and Brazil.On the other hand, for children from five years old, there is great variation in agreements. The WHO height-for-age charts showed similar patterns for the Brazilian population, while South Asian immigrants living in the Netherlands had lower height values than WHO\u2019s standard population, and the populations of Australia, Slovakia and Germany had higher height values, which indicates that this international reference does not detect stunting adequately. Regarding BMI-for-age, WHO growth charts were accurate for the Brazilian and Canadian populations, while IOTF growth charts were more accurate for the populations of Slovakia, Portugal, Italy and Poland, and CDC growth charts were accurate only for Portugal and Canada. Regarding China, Iran and Saudi Arabia, the authors suggest the use of national growth charts and, for South Africa, they point out the need for further studies to determine the most accurate international growth reference.The explanation for the international recommendation of the WHO\u2019s reference only for children under five years of age is its modeling and construction, which involved multiethnic populations in environmental and health conditions adequate for their development. Therefore, when it is applied, it presents satisfactory agreements for nutritional status assessment. The opposite is observed when the WHO growth reference is applied to children and adolescents from five years old. This is because the modeling and population used were different, resulting in divergences in nutritional status assessment in several countries, hence its use not widely indicated.In summary, the international growth charts for children and adolescents from five years old have limitations, since the differences between models and the composition of samples in the construction of growth charts did not allow an international standard for classification of nutritional status. It is difficult to obtain growth charts for international use that can go along with a long period of growth and which include genetic, cultural, socioeconomic and body composition differences of multiethnic children and adolescents who have already overcome the secular trend in height."} +{"text": "Background and Purpose: The unilateral onset and persistent asymmetry of motor symptoms are important characteristics of Parkinson's disease (PD). By using scales and wearable sensors, this study explored whether motor symptom laterality could affect non-motor symptom and gait performance.Methods: A total of 130 right-handed patients with PD were enrolled in our study and were divided into two groups according to the side of predominant motor symptom presentation by using the Unified Parkinson's Disease Rating Scale part III. We measured the non-motor symptoms with the Non-motor symptoms Scale, sleep quality with the Parkinson's Disease Sleep Scale and Pittsburgh sleep quality index, cognitive function with the Mini-mental State Examination and Montreal Cognitive Assessment, quality of life with the Parkinson's Disease Questionnaire-39, and the severity of anxiety and depression with the Hamilton Anxiety Scale and Hamilton Depression Scale, respectively. All participants underwent the instrumented stand and walk test, and gait data were collected using a set of JiBuEn gait analysis system.Results: We observed that left-dominant symptom PD patients (LPD) were associated with a greater impairment of sleep quality than right-dominant symptom PD patients (RPD). We found no difference between LPD and RPD in terms of gait performance. However, compared with the severe asymmetry RPD patients (RPD-S), severe asymmetry LPD patients (LPD-S) showed a shorter stride length and decreased range of motion of hip joints.Conclusions: In this study, LPD was associated with a more severe sleep-related dysfunction than RPD. In addition, LPD-S exhibited more gait impairments than RPD-S. Considering that motor symptom laterality may affect the non-motor symptom and gait performance, it should be taken into account when evaluating and treating PD patients. Parkinson's disease (PD) is a common neurodegenerative disease, which is mainly characterized by movement disorders, including bradykinesia, resting tremor, rigidity, and postural instability . The uniIn our research, we enrolled 130 patients from the Department of Geriatrics, Nanjing Brain Hospital Affiliated to Nanjing Medical University from October 2018 to November 2020. All patients were diagnosed with PD according to the Movement Disorder Society criteria . We exclThe following baseline data of all the participants were collected: gender, age, height, weight, and the duration of the disease. Hoehn & Yahr (H-Y) stage and UPDRS were used to assess the motor symptoms. To assess the non-motor symptoms, we used the Non-motor symptoms Scale (NMSS). Parkinson's Disease Sleep Scale (PDSS) and Pittsburgh sleep quality index (PSQI) were applied to evaluate the patients' sleep quality. Cognitive function was assessed using the Montreal Cognitive Assessment (MoCA) and Mini-mental State Examination (MMSE). The quality of life was evaluated using the Parkinson's Disease Questionnaire-39 (PDQ39). Severity of anxiety and depression were quantified by the Hamilton Depression Scale (HAMD) and Hamilton Anxiety Scale (HAMA), respectively.In our study, we used a set of JiBuEn gait analysis system to collect gait data . The accP < 0.05 indicated significant difference. Kolmogorov\u2013Smirnov test was initially used to check the normal distribution of quantitative data. Two-sample t-tests were used when both sets of data followed a normal distribution; otherwise, the Mann\u2013Whitney U-Test was used. Chi-square test was used for qualitative data. The following formula (1) was used to compute for the variability of gait parameters, and then, integrated using formula (2) , and a mula (2) , 18. We mula (2) , 20.CV indicates the coefficient of variation, and the subscripts L and R mean the left and right sides of patients, respectively.where X = , and the subscripts L and R indicate the left and right sides of patients, respectively. SL, stride length; ST, stride time; SwPT, swing phase time; StPT, stance phase time; HS, heel strike angle; TO, toe-off angle; ROM, range of motion; AJ, ankle joint; KJ, knee joint; HJ, hip joint.In our research, a total of 115 patients with PD were grouped into two groups, and their baseline data are listed in P < 0.001) and PSQI , and statistical difference was observed between LPD-S and RPD-S for PDSS . All related data were listed in In this research, non-motor symptoms including NMSS, PDQ-39, MoCA, MMSE, HAMD, and HAMA were similar in both groups. However, compared with the LPD group, the RPD group scored better in PDSS .The following spatiotemporal gait parameters were collected in our research : stride P = 0.039).Heel strike (HS) and toe-off (TO) angles were measured in our study. In addition, we evaluated the range of motion (ROM) of ankle joints (ROM-AJ), knee joints (ROM-KJ), and hip joints (ROM-HJ) in our research . We measGait symmetry data were provided in In this study, we enrolled 130 patients to assess the patients' non-motor symptoms, and explored the effect of motor symptom laterality on the gait characteristics of PD patients by using wearable devices. In-depth knowledge of both the non-motor symptoms and specific gait characteristics differences between LPD and RPD will aid in the management of PD patients.In our study, we found that LPD showed a more severe sleep-related dysfunction than RPD. This result is similar to the previous study, which indicated that left-onset PD patients suffered more daytime dozing and nocturnal hallucinations . Recent It was reported that LPD is asssociated with a higher overall non-motor impairment than RPD . This fiIt has been well-known that patients with PD demonstrated a smaller step length , 33, sloWe expanded the scope of their research and our research located this mild but significant motor impairment in short stride length and decreased hip joint movement.de novo group, and some participants had taken anti-PD drugs before. However, by stopping antiparkinsonian medication for 24 h (72 h for controlled release anti-PD drugs), we minimized the effect of drug intervention. Secondly, in this study, the assessment of the dominant hand mainly depends on the patient's self-report, rather than a systematic scale, such as the Edinburgh Handedness Inventory (Our study also has some limitations. First of all, this research was not a nventory . Finallyde novo group and longitudinal cohort, is needed to confirm these results.In conclusion, despite the limitation we stated above, we found that compared with RPD, LPD demonstrated more specific gait impairments and sleep-related dysfunction. This study exhibited an effect of motor symptom laterality on the non-motor symptoms and gait performance in PD. This small but significant effect should be considered when evaluating and treating PD patients. However, more research, especially on the The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.The studies involving human participants were reviewed and approved by Ethics committee of Nanjing Brain Hospital Affiliated to Nanjing Medical University. The patients/participants provided their written informed consent to participate in this study.LZ, SZ, and MZ conceived and designed the research. LZ and JY obtained the fundings. YB, ZW, RG, XJ, BS, and JZ collected the data. YP, PX, JD, and JY conducted the data analysis. SZ and MZ drafted the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Vaccine administration is one of the most efficient ways to control the current coronavirus disease 2019 (COVID-19) pandemic. However, the appearance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants can avoid the immunity generated by vaccines. Thus, in patients with a complete vaccine schedule, the infection by SARS-CoV-2 may cause severe, mild, and asymptomatic manifestations of the disease. In this case report, we describe for the first time the clinical symptoms of four patients from Santiago of Chile, with a complete vaccination schedule with two doses of CoronaVac (Sinovac Life Science) infected with the variant of interest (VOI) B.1.621 (Mu). They were compared with four unvaccinated patients, who had a higher prevalence of symptoms after infection compared to vaccinated patients. In the CoronaVac-vaccinated group, an 80-year-old patient who registered various comorbidities required Invasive mechanical ventilation for 28 days with current home medical recovery discharge. By contrast, in the unvaccinated group, a 71-year-old presented more symptoms with more than 45 days of Invasive mechanical ventilation, which continues to date, presenting greater lung damage than the vaccinated hospitalized patient. This first report evidence differences in the clinical symptomatology of patients vaccinated and non-vaccinated infected with the VOI B.1.621 (Mu) and suggest the protective effects of CoronaVac against this variant. The coronavirus disease 2019 (COVID-19) pandemic has left more than 4.8 million deaths around the world and 240 million infections to date. Therefore, the health authorities of all countries have implemented various protocols to prevent its spread. In this way, massive vaccination has been the most effective strategy to reduce the number of positive cases in the population and the severe clinical manifestations of the disease . HoweverThe first case of symptomatology description includes four patients with complete vaccination schedule (two doses) of CoronaVac (Sinovac Life Science) infected with the VOI Mu. The data describe two patients who are men and aged 27 and 33 years old, and two patients who are women and aged 35 and 80 years old. They become COVID-19 positive after 175, 183, 50, and 183, days respectively after the second dose of vaccination . The sec3 copies/\u03bcl), who presented several symptoms (5 copies/\u03bcl), who were asymptomatic as previously reported by our group and othesymptoms . The higptomatic . PatientA vaccinated 80-year-old patient (P1) with comorbidities, such as Diabetes type 1 (DM1), hypothyroidism, arterial hypertension, and mitral valve disease was hospitalized and admitted into the intensive care unit (ICU) for 48 days with 28 days of invasive mechanical ventilation (IMV). Her symptoms were just coughing with expectoration and shortness of breath . She is On the other hand, in the unvaccinated group, we registered a 71-year-old patient (P7) with arterial hypertension, who presented fever, dry cough, body aches, and nasal congestion. The patient remains hospitalized until the date of this report, with 58 days in the ICU and 45 days of IMV. He was admitted to the emergency department on November 6, 2021, a CCA-T was performed, showing bilateral, subpleural, and greater GGO toward the bases, which involved 18.59% of the lungs . The patThe current application of different COVID-19 vaccines has made it possible to reduce the spreading of the infection and severity of the disease in several countries . HoweverIn Chile, the most administered vaccine is the inactivated-virus-CoronaVac (Sinovac Life Sciences), approved by the ISP for emergency use . A phaseIn this report, we describe for the first time the symptomatologic scenario of four patients infected with Mu variant after 172 (P1), 50 (P2), 171 (P3), and 183 (P4) days of completing their two-doses vaccination schedule with CoronaVac vaccine. The patients P2 and P3 did not report comorbidities but reported five mild symptoms, while the P4 was asymptomatic even having the highest viral load of all patients and with a clinical history of arterial hypertension, the most common COVID-19 comorbidity that increased risk of severe disease . The unvThis is a descriptive report of the Mu variant infection in patients with a complete CoronaVac vaccine schedule. However, the information is still limited to conclude that mild symptoms are because of the complete vaccination of the patient. We suggest a direct effect of this vaccine, but it is necessary to increase the number of cases. In this way, the genetic, age, and comorbidities (among other factors of relevance) of the patients, and their potential association to the severity of the disease is also a matter of interest to be considered. In any case, our data are of great interest for future clinical investigations related to CoronaVac, its effectiveness against the new variants of SARS-CoV-2, and the direction of future public health policies aimed at eradicating the pandemic.https://www.ncbi.nlm.nih.gov/, PRJNA772359.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: The studies involving human participants were reviewed and approved by Ethical Committee of the University of Santiago of Chile (No. 226/2021) and the Scientific Ethical Committee of the Central Metropolitan Health Service, Ministry of Health, Government of Chile (No. 370/2021). Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.FER-L, CA-C, AS, EV-V, and MI: conceptualization. FER-L and RL: data curation. IH, CP, RV, SV, and MV: formal analysis. AS: funding acquisition. CB-A, AM-T, and AI-M: investigation. FER-L: methodology. DV: supervision. CB-A and RL: writing\u2014original draft. AMS, FR-L, CB-A, and RV: writing\u2014review and editing. All authors have read and agreed to the published version of the manuscript, contributed to the article, and approved the submitted version.The Laboratory of Virology had the support from the COVID-19 diagnosis in the University Laboratories Network for diagnosis tasks. The authors also thank the Rapid Assignment of Resources for Research Projects on the Coronavirus (COVID-19) , Fondecyt regular project numbers 1201664 (MI) and 1211841 (FER-L) , Fondecyt iniciaci\u00f3n grant (EV-V), and DICYT-USACH project number 021943AC (CA-C) grants. The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Adherence in the treatment of latent tuberculosis infection (LTBI) is closely related to reactivation and infection control in the population. However, there has been little research on which populations are at higher risk of loss to follow-up. The aim of this study is to investigate how the adherence of LTBI patients in the United States (US) differs by region of origin.A retrospective, observational study was conducted from 2001 to 2020. LTBI patients were identified from the Cuyahoga County Tuberculosis Clinic in Cleveland, Ohio. Only patients who were informed of the diagnosis of LTBI were included. Patients were discharged from the Tuberculosis outpatient clinic upon completion of treatment or when the physician decided to discontinue treatment. We defined loss to follow-up as a case where LTBI was diagnosed but the patient was not formally discharged. Patients whose treatment was interrupted due to side effects were not considered loss to follow-up. Odds ratios were calculated using a multivariable regression model with patients from North America as the reference group.Of 4018 LTBI patients, 1171 (28.7%) were lost to follow-up, of which 950/2314 (41.0%) were from North America. Compared with LTBI patients from North America, significantly lower loss to follow-up rates were observed for those from Middle East and North Africa 30/170 0.31-0.89), South Asia 60/692 , and Sub-Saharan Africa 69/526 . The analysis showed that a high loss to follow-up rate was observed in the patient groups from North America, Europe and Central Asia, and Latin America & the Caribbean. LTBI patients from North America had a significantly higher loss to follow-up rate than those from Middle East and North Africa, South Asia, and Sub-Saharan Africa, respectively. Further research is needed to determine how to intervene in the poorly adherent patient population, such as LTBI patients from North America, Europe and Central Asia, and Latin America & the Caribbean.All Authors: No reported disclosures"} +{"text": "In 2019, we initiated a call for papers to contribute to a Frontiers of Public Health Research Topic on the use of the Reach, Effectiveness, Adoption, Implementation, and Maintenance (RE-AIM) Framework. In part, the Research Topic was intended to celebrate 20 years of planning and evaluation using RE-AIM and underscore new research directions. More importantly, we saw the need to document how the framework continues to be adapted, evolve, and provide opportunities in emerging areas. Of key importance is speeding research-practice translation and studying the process that leads to more efficient movement of evidence-based principles, programs, and policies into sustained community and clinical practice. What follows is an overview of the excellent submissions we received that document lessons learned, adaptations, and innovative uses of RE-AIM, as well as highlight the framework's potential to advance science, quality of practice, and population health.Introduction; (b) Use of RE-AIM in Community Settings; (c) Use of RE-AIM in Clinical Settings; and (d) Emerging Directions in the Application of RE-AIM. The Introduction section kicks off the collection with a paper led, fittingly, by , on how the RE-AIM Framework has evolved over the 20 years since the seminal American Journal of Public Health article was published in 1999. This is accompanied by an insightful commentary from Dr. Kurt Stange that highlights the value of using RE-AIM and its contextual extension, the Practical, Robust, Implementation, and Sustainability Model (PRISM) to \u201cshine light on multilevel context\u201d and solve real world problems, such as advancing health equity, in a meaningful and sustained way (Stange). We round out the Introduction with two papers summarizing goals, resources, benefits of application, and future directions for RE-AIM by members of the National RE-AIM Workgroup . These papers, along with the RE-AIM website (www.re-aim.org), provides readers with current definitions, examples, and resources for applying RE-AIM.The Research Topic and resulting papers are grouped into four sections: (a) Use of RE-AIM in Community Settings and the Use of RE-AIM in Clinical Settings, respectively. Papers in these sections include international studies focused on physical activity promotion in Brazil and work environment improvements in Denmark . There are exceptional exemplars across these sections to guide readers about the application of RE-AIM when planning, implementing, and/or evaluating specific interventions intended to improve effectiveness on participant health outcomes. There are also some thoughtful manuscripts illustrating qualitative and mixed methods approaches, and one applying RE-AIM in an iterative fashion to support ongoing improvements in implementation . Finally, these sections conclude with an informative article about how to integrate an additional implementation science theory with RE-AIM constructs .The second and third sections of the Research Topic are the Emerging Directions in the Application of RE-AIM. Two studies and a commentary, focus on applications of RE-AIM (Baumann) in the areas of environmental health and health policy . They provide interesting and novel adaptations that offer opportunities for replication and further evolution of the framework. This section, and the entire Research Topic, concludes with two papers that provide extensions to RE-AIM. One integrates Proctor's et al. conceptualization of implementation outcomes , and the other integrates PRISM and RE-AIM factors to advance sustainability and health equity . The papers in this section are strong examples of how RE-AIM can be conceptualized, re-conceptualized, and expanded to address and resolve. As called out in the Introductory commentary by Stange, these papers astutely address the wicked problems affecting the health and equity in our society by taking the path that creates and supports a high, robust, and sustained level of public health around the world.Our Research Topic concludes with a collection of papers that reflect It is our hope that this collection of papers provides concrete examples and guidance about how RE-AIM can be used to advance science and improve the public health impact in our communities, both nationally and internationally.All authors contributed to the conceptualization of the manuscript and its content. All authors contributed to the full manuscript as well as reviewed and approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "In non-randomized studies (NRSs) where a continuous outcome variable is assessed at baseline and follow-up, it is common to observe imbalance of the baseline values between the treatment/exposure group and control group. This may bias the study and consequently a meta-analysis (MA) estimate. These estimates may differ across statistical methods used to deal with this issue. Analysis of individual participant data (IPD) allows standardization of methods across studies. We aimed to identify methods used in published IPD-MAs of NRSs for continuous outcomes, and to compare different methods to account for baseline values of outcome variables in IPD-MA of NRSs using two empirical examples from the Thyroid Studies Collaboration (TSC).For the first aim we systematically searched in MEDLINE, EMBASE, and Cochrane from inception to February 2021 to identify published IPD-MAs of NRSs that adjusted for baseline outcome measures in the analysis of continuous outcomes. For the second aim, we applied analysis of covariance (ANCOVA), change score, propensity score and the na\u00efve approach (ignores the baseline outcome data) in IPD-MA from NRSs on the association between subclinical hyperthyroidism and depressive symptoms and renal function. We estimated the study and meta-analytic mean difference (MD) and relative standard error (SE). We used both fixed- and random-effects MA.Ten of 18 (56%) of the included studies used the change score method, seven (39%) studies used ANCOVA and one the propensity score (5%). The study estimates were similar across the methods in studies in which groups were balanced at baseline with regard to outcome variables but differed in studies with baseline imbalance. In our empirical examples, ANCOVA and change score showed study results on the same direction, not the propensity score. In our applications, ANCOVA provided more precise estimates, both at study and meta-analytical level, in comparison to other methods. Heterogeneity was higher when change score was used as outcome, moderate for ANCOVA and null with the propensity score.ANCOVA provided the most precise estimates at both study and meta-analytic level and thus seems preferable in the meta-analysis of IPD from non-randomized studies. For the studies that were well-balanced between groups, change score, and ANCOVA performed similarly. In non-randomized studies (NRS) that assess a continuous outcome of interest at baseline and follow-up, baseline values between treatment or exposure and control group may differ significantly. Ignoring this imbalance in the analysis may confound the estimated study effect . LikewisThe most common methods are analysis of covariance (ANCOVA) and the change score. These methods are both based on a linear regression model , 4. ANCOAnother method that may account for baseline imbalance in study analysis is the propensity score, also called inverse probability weighing, which accounts for baseline imbalances by assigning weights to each participant. In this method, the researcher applies a linear regression model with follow-up values as outcome and each participant is weighted for the conditional probability of being treated or exposed, given the baseline outcome. Weights are calculated as the inverse probability of being treated/exposed given baseline outcome values, under the assumption of no unmeasured confounders that may affect the estimate and the causal effect of the exposure , 8. ThisAnother issue is that the pooled estimate obtained from the MA of NRSs with biased estimates due to ignoring imbalance at baseline in the study statistical analysis may also be biased, as well as less efficient , 10. If Our first aim was to identify the statistical methods IPD MA of NRSs used to deal with continuous outcomes assessed at baseline and follow-up. Our second aim was to compare the impact of the above methods in the study and meta-analytic estimates in two empirical examples of IPD-MA of NRSs.https://rayyan.qcri.org/) and removed duplicates. Two authors (LS and LW) independently screened citations by title and abstract against predefined eligibility criteria. The same two authors reviewed the full text of all selected records. They resolved disagreements by discussion and, if needed, consulted a third author (CDG) to reach consensus. From each eligible IPD-MA, we extracted the following information: number of included cohorts/studies; number of participants; clinical field; assessment of potential outcome baseline imbalance between groups; assessment of the correlation between baseline and follow-up outcome data; primary statistical method that accounted for baseline outcome data, and eventual method used in a secondary analysis. We piloted an electronic data extraction form that was used by the two reviewers to extract information of interest from included publications.To identify the various statistical methods, we systematically reviewed published IPD-MAs of NRSs that analyzed continuous outcomes and used baseline outcome data in the analysis. We built the search strategy with the help of a medical librarian. We searched Medline (PubMed), Embase (Ovid), and CENTRAL (Cochrane Library) from inception to February 2021 using the key terms listed in the For our second aim we used data from the Thyroid Studies Collaboration (TSC): (1) Wildisen et al. assessed the association between subclinical hyperthyroidism (exposure) and depressive symptoms (outcome) , and (2)2; values lower than 60 mL/min/1.73m2 indicate deteriorated renal function. eGFR was calculated with the four-variable Modification of Diet in Renal Disease formula when it was not in the original source data.We included cohorts with available data on the outcome of interest at baseline, at first available follow-up, and with thyroid status at baseline (measured TSH). Depressive symptoms were measured on a validated depression scale in the Beck Depression Inventory (BDI). BDI scales go from 0 to 63; higher values indicate more symptoms of depressive symptoms . We meast-test. We verified the data were normally distributed. For each cohort, we also calculated the correlation coefficient between baseline and follow-up outcome data. Then we executed a two-stage IPD-MA. In the first stage, we estimated the study-specific mean difference (MD) of the outcome between participants with subclinical hyperthyroidism and euthyroid participants and, to measure the precision of the estimates, the relative standard error (SE). We obtained study estimates from ANCOVA, change score, and propensity score. For comparison, we also applied the na\u00efve approach, which model follow-up outcome data and ignores baseline outcome data. Na\u00efve model has been showed to produce biased estimates in case of the presence of baseline imbalance .We analyzed only participants whose baseline and follow-up data were both available. We also collected data on age and sex for each cohort. We calculated the mean and standard deviation (SD) of the continuous outcomes at baseline and follow-up in each cohort study and assessed statistical baseline imbalances between groups with the mbalance . Since wOur initial search yielded 2,611 unique citations, which we scrutinized for eligibility. 2 = 0.56) compared to that from ANCOVA (\u03c42 = 0.13) and null for the propensity score and na\u00efve approach between baseline and follow up outcome . Results2 = 3.18); it was lower for ANCOVA (\u03c42 = 1.00) and the propensity score (\u03c42 = 0.16) and it was null for the na\u00efve approach. In both fixed and random effects, pooled results from propensity score showed less renal deterioration in the exposure group compared to the control group, while the other methods showed results in the other way round , the change score was the most common statistical method, followed by ANCOVA\u2014an unexpected finding because Cochrane recommends using ANCOVA to incorporate baseline outcome data in meta-analysis . A recenOverall, our findings are consistent with previous studies that suggested ANCOVA was most precise and better accounted for baseline imbalance between groups , 2. Our Our study had three limitations. First, it did not assess the effect of the methods in both aggregate and IPD datasets. Second, for ANCOVA we assumed a linear confounding effect of baseline outcome data. However, association with follow-up may not be linear and a spline term may be included in the model to allow for potential non-linear confounding effect. Third, we only explored the effect of the methods in empirical examples; assessment via simulation studies may be further conducted.For non-randomized studies that were well-balanced between groups, change score and ANCOVA performed similarly, but ANCOVA provided the most precise estimates at both study and meta-analytic level. In consistency with studies that showed biased estimates using change score in not randomized studies, we recommend using ANCOVA in meta-analyses of individual patient data from non-randomized studies.https://www.thyroid-studies.org/. Requests to access these datasets should be directed to Cinzia Del Giovane, cinzia.delgiovane@biham.unibe.ch.The data analyzed follow restrictions of each included study cohort. For more information, see the link Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.LS, LW, and CDG have full access to all of the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis and had the final responsibility for the decision to submit for publication. LS, LW, CDG, and NR: concept and design. LS, LW, CM, and CDG: acquisition, analysis, or interpretation of data. LS and CDG: drafting of the manuscript and statistical analysis. CM, DB, AC, JG, WE, ST, RW, JJ, LF, GC, B\u00c5, LC, RP, MI, WO, BV, HV, JS, JW, RD, SB, MI, and NR: critical revision of the manuscript for important intellectual content. LS, LW, CM, DB, AC, JG, WE, ST, RW, JJ, LF, GC, B\u00c5, LC, RP, MI, WO, BV, HV, JS, JW, RD, SB, MI, and NR: administrative, technical, or material support. CDG: supervision. All authors have read and approved the final manuscript.www.thyroid-studies.org) was supported by grants from the Swiss National Science Foundation (SNSF 320030-172676 and 32003B_200606 both to NR). The Busselton Health Study had no financial support to disclose. The Cardiovascular Health Study (CHS) was supported by contracts HHSN268201200036C, HHSN268200800007C, HHSN268201800001C, N01HC55222, N01HC85079, N01HC85080, N01HC85081, N01HC85082, N01HC85083, N01HC85086, 75N92021D00006, and grants U01HL080295 and U01HL130114 from the National Heart, Lung, and Blood Institute (NHLBI), with additional contribution from the National Institute of Neurological Disorders and Stroke (NINDS). Additional support was provided by R01AG023629 from the National Institute on Aging (NIA). A full list of principal CHS investigators and institutions can be found at CHS-NHLBI.org. The European Prospective Investigation of Cancer (EPIC)-Norfolk study was supported by research grants from the Medical Research Council UK and Cancer Research UK. The Health, Aging and Body Composition study was supported by NIA Contracts N01-AG-6-2101; N01-AG-6-2103; N01-AG-6-2106; NIA grant R01-AG028050 and NINR grant R01-NR012459. This research was funded in part by the Intramural Research Program at the NIA. The InChianti study was supported as a target project ICS 110.1jRS97.71 by the Italian Ministry of Health, and in part by the US NIA, contracts 263-MD-9164-13 and 263-MD-821336. The Tr\u00f8ndelag Health Study (HUNT) is a collaborative effort of HUNT Research Center , the Norwegian Institute of Public Health, Central Norway Regional Health Authority and the Tr\u00f8ndelag County Council. Thyroid function testing in the HUNT Study was financially supported by WallacOy . The Leiden 85-plus study was partly funded by an unrestricted grant from the Dutch Ministry of Health, Welfare and Sports (1997\u20132001). The original PROSPER study was supported by an unrestricted, investigator-initiated grant from Bristol-Myers Squibb. The Rotterdam Study was funded by the following: Erasmus MC and Erasmus University, Rotterdam, the Netherlands; the Netherlands Organisation for Scientific Research (NWO); the Netherlands Organisation for the Health Research and Development (ZonMw); the Research Institute for Diseases in the Elderly (RIDE); the Ministry of Education, Culture and Science; the Dutch Ministry for Health, Welfare and Sports; the European Commission (DG XII); and the Municipality of Rotterdam. The Radiation Effects Research Foundation (RERF), Hiroshima and Nagasaki, Japan, was a public interest foundation funded by the Japanese Ministry of Health, Labour and Welfare (MHLW) and the US Department of Energy (DOE). This publication was supported by RERF Research Protocol A5\u201313. SHIP was part of the Research Network of Community Medicine at the University Medicine Greifswald, Germany (www.communitymedicine.de), which was funded by the German Federal State of Mecklenburg\u2013West Pomerania. The BELFRAIL study was funded by an unconditional grant from the Fondation Louvain. The Fondation Louvain was the support unit of the Universit\u00e9 Catholique de Louvain in charge of developing education and research projects of the university by collecting gifts from corporate, foundations and alumni. The Brazilian thyroid study was supported by an unrestricted grant from S\u00e3o Paulo State Research Foundation (Fundac\u00e3o de Amparo a Pesquisa do Estado de S\u00e3o Paulo) Grant 6/59737-9. The Prevention of Renal and Vascular End-Stage Disease (PREVEND) study has been made possible by grants from the Dutch Kidney Foundation: (E.033).The work from the Thyroid Studies Collaboration (TSC, The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The views of the authors do not necessarily reflect those of the two governments.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Background: Several studies have reported an association between temporomandibular disorder pain (TMD-P) and emotional disorders in children and adolescents. However, no studies have reported if self-reported TMD-P in Saudi Arabia is associated with psychosocial symptoms. Therefore, the current study aimed to evaluate the association between self-reported TMD-P with depression, anxiety and somatic problems in children and adolescents in Saudi Arabia. The hypothesis was that there is an association between self-reported TMD-P and psychological symptoms among children and adolescents.Materials and Methods: The included participants were randomly selected boys and girls aged between 10 and 18 years, with a mean (SD) age of 14.0 (2.3) years. Out of 633 children and adolescents that were invited to participate, 509 voluntarily agreed to participate, and 466 completed all questionnaires. The questionnaires included items retrieved from the Youth Self Report (YSR) and Axis II of the Research Diagnostic Criteria for TMD (RDC/TMD) besides demographic data, medical history, and presence of oral parafunctions. To assess the presence of self-reported TMD-Pain, each participant was verbally asked two validated questions regarding the presence of TMD-P and dysfunction (2Q-TMD).Results: Self-reported TMD-P in children and adolescents was significantly associated with anxiety, depression, somatic symptoms, and social problems (P < 0.0001). Further, the frequencies of anxiety, depression, and somatic disorders were more evident among children and adolescents who suffered from TMD-P (P < 0.0001). The odds of reporting TMD-P in children and adolescents was 1.4 times for border line and clinical diagnosis scores for anxiety and withdrawal depression domains, and 2.6 times for the somatic symptoms' domains. However, in the multiple regression model after controlling for possible confounders, only somatic symptoms and social scores were significant. Moreover, self-reported TMD-P was twice as prevalent among girls compared to boys.Conclusion: This study reports a significant association between psychosocial burden and presence of self-reported TMD-Pain, with a stronger impact on girls than boys. There were significantly higher number of participants with self-reported TMD-P reporting a poor oral and general health. In addition, self-reported TMD-P was higher among those with borderline and clinically diagnosed anxiety/depression scores. Based on this finding, the current study supports that an early approach and recognition of children and adolescents with anxiety, depression, somatic symptoms, and TMD problems. This could result in a lesser burden for these children and adolescents both in regard to pain and psychosocial implications with increased quality of life. It is well-documented that pain disorders, such as the pain in orofacial or temporomandibular joint (TMJ) regions, are comorbid with biopsychosocial factors \u20133. TempoTMD-P can be comorbid with somatic painful complaints such as headache, neck pain, and back pain \u201312. Non-Anxiety can be defined as a person's expected emotional condition in certain situations . It is cIn Saudi Arabia, it has been shown that more than half of adolescent boys have at least one emotional disorder . Other sThis cross-sectional study was conducted between January 2014 and March 2014 and was part of a project with several studies carried out in the city of Jeddah, Saudi Arabia , 15, 23.The only inclusion criterion for the participating children and adolescents was being at the age between 10 and 18 years. To obtain results that could be generalizable the present study did not have any exclusion criteria; thus, all plausible participants were invited. However, the participants of the current study were randomly selected from different schools in Jeddah city. This randomization process was conducted as follows:The city of Jeddah was divided into the five predefined regions: North, South, East, West, and Central.The inclusion of schools was based on a predefined set of schools, as clustered by the ministry of education, in order to obtain a representative sample of the city of Jeddah.The educational system in Saudi Arabia is based on single-sex schools .www.randomization.com) by one of the researchers (NC) who did not participate in data collection.Two boy-schools and two girl-schools were randomly selected from each region. This randomization was performed with a computer-based application (One class from each school (having on average 30 pupils) was also randomly selected using a simple sampling method. This simple sampling was done by using a bucket with the titles of the school classes from the specific school from which a dental assistant not participating in the data collection drew one class.As this study is part of a larger project, this paper only presents the Axis II of the Research Diagnostic Criteria for TMD (RDC/TMD). The clinical diagnoses, based on Axis I of the RDC/TMD, are presented in another study . Due to One day before their clinical examination, all girls and their parents received information about the purpose of the study and a brief explanation of the questionnaires they were asked to fill in. On the day of clinical examination (Axis I of the RDC/TMD), the participating girls were examined in their schools, during the ordinary school day, using a mobile dental chair in the nurse's room. Also, the girls completed the official Arabic version of the YSR .One appointment was offered to each presumable participant. The boys were accompanied by a parent/guardian to the clinic. To reduce the risk of parent/guardian interference the parents/guardians were asked to wait outside the clinical room. However, if the parent/guardian insisted to attend together with their son, they were asked to remain passive during the entire session. The participating boys were examined at a dental clinic, at the primary health care center of that region.All boys were examined according to Axis I of the RDC/TMD. Their parents received information about the purpose of the study and a brief explanation of the questionnaires they were asked to fill in. This questionnaire is the Arabic version of the YSR .) the presence of TMD-P, and (2) the presence of jaw dysfunction once a week or more often [Do you have pain in the temple, face, temporomandibular joint, or jaws once a week or more?,\u201d while the second question was \u201cDo you have pain when you open your mouth wide or chew once a week or more?.\u201d If the participant marked \u201cyes\u201d to one or both of the two questions was regarded as TMD-P. Thus, the participants were categorized into two groups: TMD-P and no TMD-P.To assess the presence of self-reported TMD-P, each participant was verbally asked two validated questions regarding that is reliable and used for children and adolescents [The RDC/TMD is a dual diagnostic tool , Graded Chronic Pain Scale (GCPS), and Jaw Disability Checklist (JDC).The The SCL-90-R is not validated for children and adolescents younger than 13 years of age , therefoThe data obtained from GCPS and JDC was presented elsewhere .This Arabic version of YSR is validated and reliable and was The YSR contains two main domains including 112 problem statements . The firThis domain assesses emotional and behavioral functioning that consists of 109 statements. Those statements explicate five major clusters: (1) anxiety; (2) depression; (3) somatic complaints; (4) aggressive disorders; and (5) social and attention problems. These major clusters are grouped into three subscales: (a) broad-band internalizing and externalizing; (b) eight narrow-band syndromes; and (c) DSM-oriented scales .This study focuses on the first three narrow-band syndromes, which are: (1) Anxious/Depressed; (2) Withdrawn/Depressed; (3) Somatic Complaints. The other five narrow-band syndromes, as well as the DSM-oriented scales, are already presented in a previous study . The YSRTM version 9.1, Burlington, VT, USA) was used to calculate and present percentiles and T-scores for all subscales and syndromes. The normal T-score range for all syndromes is between 50 and 64, the borderline clinical T-score range is between 65 and 69, while the clinical T-score range is between 70 and 100 [Due to cultural considerations, three statements about sexual problems were removed in the current study. This was also done in a previous study regarding TMD-P in an adult cohort in Saudi Arabia . A licen and 100 .TM version 9.1, Burlington, VT, USA) was used for this domain to calculate and present percentiles and T-scores for all activities and social competencies. The normal T-score range for all syndromes is between 36 and 65, the borderline clinical T-score range is between 32 and 35, while the clinical T-score range is between 20 and 31 [This domain measures the social competence and physical activities that comprises of seven statements covering three areas: (1) social relations; (2) physical activities; and (3) the mean of self-reported academic performance . The samSample size calculation was done using G-power (Version 3.1.9.3). Based on a previous study , resultsp < 0.0001), so Mann-Whitney U tests were used to determine if there were any significant differences in psychosocial scores based on self- reported TMD pain status. Logistic regression models were performed to predict which variables were determinants of self-reported TMD-P status at 0.05 level of significance.Statistical analyses were performed using the statistical software: IBM SPSS Statistics . The dependent variable was the self- reported TMD-P; categorized as a dichotomous variable no/yes. Demographic data were categorical and included gender, nationality, school grade, parental income, and occupation. Psychosocial scores included Anxiety/Depression score, Withdrawal/Depression score, Somatic Complaints score, Activity Score, and Social Score. Scores were not normally distributed and Withdrawal/Depression score (by 1.95 scores) compared to the age group 10\u201313 years. Girls and boys were only different in their Social scores, as girls had lower Social score by 3.88 .P < 0.0001) , being a female , and higher somatic complaints and social problems scores were significantly associated with the self- reported TMD-P (Model 0.0001) .Since pain is a subjective feeling affected by the bio-psychosocial model, our study aimed to evaluate the associations between self-reported TMD-P and depression, anxiety, and somatic problems in children and adolescents. The results showed that children and adolescents with self-reported TMD-P reported higher scores for anxiety, depression, and somatic symptoms compared to children and adolescents with no TMD-P complaint. Similar to our results, previous studies have reported that internalizing emotional problems such as depression and anxiety are associated with TMD signs and symptoms and a TMD diagnosis , 36, 37.Concerning anxiety, depression, and somatic diagnoses, our study showed that a higher proportion of children and adolescents with TMD-P reported Anxiety/Depression than other domains. A recent longitudinal study suggested that TMD-P in children and adolescents was increased three times in adulthood . Taken tThe present study showed that self-reported TMD-P was nine times more frequent among those with a confirmed RDC/TMD pain diagnosis. The current results corroborate previous results of a significant association between self-reported TMD-P and the presence of confirmed TMD diagnosis , 10. HowConcerning somatic complaint, our study showed that TMD-P is significantly associated with somatic disorders. Similarly, other studies reported headache, backache, and stomachache in children and adolescents with TMD problems . When orMoreover, the present study found a significant association between self-reported TMD-P and female sex. Self-reported TMD-P was twice as frequent among girls than boys. Similarly, Nilsson and co-workers 2001 found a higher prevalence of TMD-P among adolescents girls than adolescents boys . Other sThis study has numerous strengths. One strength is data collection through the rigor randomization process. Hence, this process would permit the generalizability of the results to the residents of Saudi Arabia. The use of two validated questions (2Q-TMD) is another strength of our study. Also the use of the self-reported YSR in the current research is another strength since it is a valid and reliable instrument . MoreoveOne could argue that the unequal numbers between boys and girls, with a higher number of dropouts among the boys, can be seen as a limitation of this study. This was probably due to the study set-up with different settings for boys and girls. The girls were examined in the school nurse's room using a mobile dental chair which made the examination very accessible. The boys, on the other hand, had to go to a dental clinic at their primary health care center, which could explain the higher number of dropouts. This difference could not be avoided due to the nature of the school system in Saudi Arabia.We concluded that there is a significant association between psychosocial problem and presence of TMD-P in children and adolescents in Saudi Arabia. There was a significantly higher number of children with poor oral and general health the reported TMD-P. Based on this finding, an early approach and recognition of children and adolescents with anxiety, depression, somatic symptoms and TMD problems could result in a lesser burden for them and in turn increase quality of life.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Local Ethical Committee at the Department of Medical Study and Research, Ministry of Health, Jeddah, Saudi Arabia. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.AA-K, BH-M, ME, and NC: conceptualization. AA-K, ME, and NC: methodology and validation. AA-K, DM, SA, NC, and ME: software. AA-K, DM, SA, and NC: formal analysis. AA-K: investigation and funding acquisition. AA-K, DM, SA, BH-M, ME, and NC: resources and writing (review and editing). AA-K, DM, and SA: data curation. AA-K and DM: writing . AA-K and NC: visualization and project administration. BH-M, ME, and NC: supervision. All authors contributed to the article and approved the submitted version.The current study was financially supported by a grant from Ministry of Health, Saudi Arabia.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Considering the physical, and psychological impacts and challenges brought about the coronavirus disease 2019 (COVID-19), art therapy (AT) provides opportunities to promote human health and well-being. There are few systematic analysis studies in the fields of AT, which can provide content and direction for the potential value and impact of AT. Therefore, this paper aims to critically analyze the published work in the field of AT from the perspective of promoting health and well-being, and provides insights into current research status, hotspots, limitations, and future development trends of AT. This paper adopts a mixed method of quantitative and qualitative analysis including bibliometric analysis and keyword co-occurrence analysis. The results indicate that: (1) the current studies on AT are mostly related to research and therapeutic methods, types of AT, research populations and diseases, and evaluation of therapeutic effect of AT. The research method of AT mainly adopts qualitative research, among which creative arts therapy and group AT are common types of AT, and its main research populations are children, veterans, and adolescents. AT-aided diseases are trauma, depression, psychosis, dementia, and cancer. In addition, the therapeutic methods are mainly related to psychotherapy, drama, music, and dance/movement. Further, computer systems are an important evaluation tool in the research of AT; (2) the future development trend of AT-aided health and well-being based on research hotspots, could be focused on children, schizophrenia, well-being, mental health, palliative care, veterans, and the elderly within the context of addressing COVID-19 challenges; and (3) future AT-aided health and well-being could pay more attention to innovate and integrate the therapeutic methods of behavior, movement, and technology, such as virtual reality and remote supervision. In December 2019, a novel coronavirus disease 2019 (COVID-19) broke out in Wuhan, China. With the rapid spread of the new crown virus, the World Health Organization (WHO) declared the COVID-19 epidemic as a public health emergency of international concern on 30 January 2020 . FurtherAmong the many non-drug treatments, art therapy (AT) is a comprehensive treatment that uses psychotherapy and artiAT is a valuable way of expression, and art shows a great promise in the direction of a sustainable future . Art-basThis paper adopts a mixed research method, which comprises a bibliometric analysis to quantitatively examine AT-aided health and well-being, identifying significant research structure and research topics, and a follow-up qualitative review to reveal the content of different AT research themes. In the quantitative data analysis, histograms, pie charts, and visual maps of keyword co-occurrence analysis in VOSviewer are used to analyze the current research status and research hotspots of AT. Bibliometric analysis is widely used to analyze the published literature in a particular field, which helps to evaluate the trend of research activities over time . AlthougScienceDirect is the fIn ScienceDirect full-text database, the research on the AT-aided health and well-being, which began in year 1946, encompasses a total of 799 published articles in three quarters of a century, spanning 75 years from 1946 to September 2021. These include including 191 articles from 1946 to 1999 and 608 articles from 2000 to 2021, which are shown in chronological order in In ScienceDirect full-text database, the 799 articles on AT-aided health and well-being are published in 139 academic journals. As shown in As shown in The overlay visualization diagram generated by VOSviewer is shown in As shown in In the Network Visualization diagram generated by VOSviewer, the more frequently keywords appear in the article, the larger the circle in the diagram and the more closely they are associated with the central theme. Since the VOSviewer software does not automatically merge the repeated synonymous keywords during the co-occurrence analysis of keywords, such as \u2018art-therapy\u2019, \u2018art-th\u00e9rapie\u2019, and \u2018art therapy\u2019; \u2018drama therapy\u2019 and \u2018psychodrama\u2019; and \u2018creative\u2019 and \u2018creativit\u00e9\u2019. Therefore, when classifying topics based on Network Visualization diagram, synonymous keywords are manually homogenized. It can be seen from In terms of the distance from each cluster to the center of AT in Based on the keyword co-occurrence analysis results of VOSviewer, keywords that are most closely related to the AT theme are selected for further qualitative analysis in following Based on the results of the above quantitative analysis, this paper qualitatively summarizes the research status of AT-aided health and well-being in ScienceDirect full-text database from the following six thematic categories that are in line with Qualitative research is the main research method of AT ,67,68. IThe types of AT-aided health and well-being mostly include CAT and group AT. CAT is a term used in therapeutic arts disciplines , which cCAT has a certain potential in relieving psychological, behavioral, physiological, and other related diseases. It has been widely used to study military groups. Group AT has numerous benefits for mothers and children, such as relieving symptoms of depression, fear, anxiety and mood disorders, and improving patients\u2019 subjective well-being and quality of life. The research population of AT-aided health and well-being chiefly focusses on children, veterans, and adolescents.AT-aided health and well-being with children as the research population largely adopt a mixed/hybrid research method integrating experiment, interview, questionnaire, and case studies ,115,116.The research of AT on the veterans group mainly adopts the case study method ,36,125. The majority of AT-aided health and well-being on adolescents use case analysis research method ,126,127,AT-aided diseases are largely focused on five aspects: (1) trauma and PTSD; (2) depression; (3) schizophrenia and psychosis; (4) Alzheimer\u2019s disease and dementia; and (5) cancer.AT-aided health and well-being for trauma and PTSD mostly conducted via case study research method ,14,125. AT-aided health and well-being on depression primarily implement experimental research methods with scales ,102,133,AT-aided studies principally employ case study research methods ,141,142 Case studies were predominantly used in the AT-aided disease research to treat Alzheimer\u2019s disease and dementia ,148,149.AT-aided cancer treatment studies have been conducted via a mixed research method using experiment, interview, and questionnaire ,152,153.Breast cancer is the disease that has been paid the most attention regarding the use of AT to treat cancer. The \u2018Coping Resources Inventory\u2019 is a commonly used assessment method, in which quality of life, fatigue, and subjective well-being are commonly used assessment elements in the process of AT treating breast cancer patients. It is able to explain and help with understanding the female image of breast cancer patients ; and proThe therapeutic methods of AT are twofold: psychology, and artistic creation. In addition to the drawing mentioned in Psychotherapy is a biopsychosocial method in the process of AT . Art psyDrama is a friendly art form , as an aMusic therapy provides opportunities to treat behavioral, psychological, and cognitive disorders . It is oD/MT is a new field of CAT , which lThe computer system acts as an evaluation tool for AT-aided health and well-being. Based on the quantitative and qualitative analysis results of this paper, it can be seen that group AT, children, and schizophrenia are the key themes of AT-aided health and well-being. There was consensus in the reviewed literature that AT is able to improve the quality of life and well-being of patients with Alzheimer\u2019s disease, dementia, and breast cancer. Overall, the qualitative analysis results also show that well-being and mental health are closely related to AT.The research hotspots of AT-aided studies from 2015 to 2021, primarily revolve around therapeutic methods and population. The four therapeutic methods of psychotherapy, drama, music, and D/MT all bring potential value to improve mood and psychological disorders. AT is also often associated with palliative care to explore the therapeutic effect. In addition to alleviating the symptoms of patients, AT-based palliative care interventions help patients and their relatives to improve their sensory, emotional, cognitive, and spiritual experiences , elevatiAmong the hot research population on AT, in addition to children, adolescents and veterans, the elderly has also become the key research population of AT. The positive therapeutic effect of AT on the elderly is mainly reflected in helping them prevent cognitive decline , and impFurthermore, the results of AT-aided health and well-being indicate that among the latest research keywords in 2021, the research content of AT also revolves regarding the keyword of COVID-19. In the context of the COVID-19 pandemic, Hass-Cohen et al. studied The results reveal that the primary therapeutic methods of AT-aided health and well-being include psychotherapy, D/MT, drama, music, and drawing. Drawing includes self-portrait, mandala, PPAT, and bridge drawing as its approaches. In addition, AT-aided cognitive behavior is an effective method to promote health, which focuses on the therapeutic effect of facilitating behavior with psychodrama ,195, andIn the light of mega digital era, AT needs to utilize continuously evolving emerging technologies to make an effective intervention in its process. The application of emerging technology includes digital technology and remote technology, such as VR, digital phototherapy technology, computer technology, and telemedicine technology in the current state of AT-aided health and well-being. Mihailidis et al. argue thWith the use of mixed research method, this paper summarizes the current situation, hot spots, deficiencies, and future research trends of the practical application of AT from both quantitative and qualitative aspects in promoting health and well-being, which provides specific content and direction for the potential practical value of AT. This paper has three main contributions: (1) this paper is the first mixed research method to incorporate AT articles of 75 years (3/4 century) by using visual keyword co-occurrence. This comprehensive research result has reference value for AT researchers, educators, and healthcare practitioners and can provide pathways for information and communications technology (ICT) development for information visualization software suppliers. (2) This paper is the first attempt to use bibliometric analysis which includes keyword co-occurrence analysis to classify AT-aided study status in ScienceDirect full-text database from 1946 to September 2021. VOSviewer, a tool for visualizing bibliometric graphs, is used for keyword co-occurrence analysis. It is able to gain insights into AT related topics through visualized keyword maps. With the help of VOSviewer, the systematically mixed quantitative and qualitative analysis summarizes the research categories, research hotspots, and research deficiencies of AT, which provide a reliable research method for the future study of AT. (3) This paper finds that the research status of AT-aided study primarily includes research methods, types, populations, diseases, therapeutic methods, and evaluation of six themes. The hot keywords of AT-aided health and well-being mainly focus on group AT, children, schizophrenia, well-being, mental health, palliative care, veterans, and the elderly, which may be used as a basis to analyze the background of the COVID-19 in the future AT-aided health and well-being. In addition, in spite of the gaps and shortcomings in the exploration of AT in behavior, movement, and technology, the integration and innovation of behavior, movement, and technology in the field of AT is a multidimensional breakthrough in promoting health and well-being. However, the research in this paper has certain limitations. Different researchers may use different keywords to express the same meaning due to differences in terms used by individuals, such as \u2018older adults\u2019 representing \u2018the elderly\u2019, which could affect the retrieval effect. In addition, VOSviewer software cannot automatically homogenize repeated synonymous keywords when performing keyword co-occurrence analysis. As such, manual homogenization of keywords may lead to slight deviations in data analysis. Further, this paper only conducts bibliometric analysis on a single ScienceDirect database, and does not extend the search data of AT to multiple databases in different fields such as health, society, and art. Future research could be based on the research status and research limitations of AT to systematically conduct visual analysis from multiple databases, such as Web of Science and Scopus, in order to create more application values for various social situations, such as COVID-19. Further, based on the research in the fields of behavior, movement, and technology, the therapeutic effect of AT could be analyzed more comprehensively and concretely from the aspect of promoting health and well-being."} +{"text": "In \u201cA COVID-19 Pandemic Artificial Intelligence\u2013Based System With Deep Learning Forecasting and Automatic Statistical Data Acquisition: Development and Implementation Study\u201d :e27806), the authors noticed two errors.In the originally published manuscript, authors Cheng-Sheng Yu and Shy-Shin Chang should have been denoted as having contributed equally to the paper but were not. A footnote clarifying equal contribution has now been added to each of the aforementioned authors.Additionally, in the originally published manuscript, the Acknowledgments section was omitted. In the corrected version, a new Acknowledgments section has been added with the following statement:This study is supported by the Ministry of Science and Technology Grant (MOST 110-2314-B-038-025) and Higher Education Sprout Project by the Ministry of Education (MOE) in Taiwan (DP2-110-21121-01-A-09). No funding bodies had any role in study design, data collection and analysis, decision to publish, or preparation of the article.The correction will appear in the online version of the paper on the JMIR Publications website on July 9, 2021, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "The aggregation of the protein \u03b1-synuclein (aSyn) is the pathological hallmark of the group of neurodegenerative disorders, collectively known as synucleinopathies. These include Parkinson's disease (PD), PD-Dementia, Dementia with Lewy Bodies (DLB), and Multiple Systems Atrophy (MSA). While all of these neurodegenerative disorders present with distinctive clinical features, they all converge in one pathological characteristic: intracellular aSyn aggregation into Lewy Bodies .aSyn is typically degraded by both the lysosome and the proteasome , lysosomal cathepsin D variants associated with neurodegenerative disorders were analyzed , no. 4. et al.) , the stu et al.) , no. 2.Ray et al. revises the importance of aSyn structure and mutations on the biophysics of its aggregation, cell autonomous pathobiology, as well as spreading of disease . Consequences of two common familial-associated mutations (A30P and A53T) were evaluated on protein aggregation and locomotor behavior in a C. elegans model . Furthermore, Fouka et al. summarizes potential treatment strategies aiming at preventing both protein aggregation and cell-to-cell propagation via utilization of antibodies against aSyn . Moreover, lysosomal as well as mitochondrial pathways are highlighted for therapeutic strategies via calcium and iron modulation among others .The structural properties and posttranslational modifications (PTMs) of aSyn play an important role in toxicity and its seeding capacity (Oliveira da Silva and Liz) . Reinforcing the role of aSyn in the adult hippocampus, an increase in the number of early stage neuronal progenitors in a human aSyn transgenic mouse model was shown , no. 9, et al.) , no. 7, and Liz) , no. 8. et al.) , no. 10. et al.) .Grozdanov and Danzer) . A better comprehension of aSyn function and structure within the GIT will be crucial to understand its role in the enteric nervous system and its role in spreading from the gut to the brain , no. 11.In summary, the articles within this Research Topic provide an overview of intracellular mechanisms that mediate the conversion from physiologic to toxic aSyn conformations, the intracellular consequences of toxic aSyn, as well as spreading mechanisms that accelerate pathology in nearby cells and other tissues . A betteFZ provided the figure. All authors contributed to the manuscript.FZ is supported by the German Research Foundation (DFG), grant number 125440785 - SFB 877 (project B11) and the Interdisciplinary Center for Clinical Research (IZKF) at the University Hospital of the University of Erlangen-Nuremberg . Additional support for BW came from the Bavarian Ministry of Science and the Arts in the framework of the ForInter network, the German Research Foundation, DFG WI 3567/2-1 and 270949263/GRK2162, and the IZKF advanced project E30. GC is supported by the Parkinson's Foundation PF-JFA-1949 and R01 NS117750/NS/NINDS NIH HHS/United States.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Funding statement appears below.In the original article, funder FCT, 2020.07675.BD was not included. The corrected This work was supported by the Research Center of the Portuguese Oncology Institute of Porto (project no. PI86-CI-IPOP-66-2017); by European Investment Funds by FEDER/COMPETE/POCI-Operational Competitiveness and Internationalization Program, and National Funds by FCT-Portuguese Foundation for Science and Technology under projects UID/AGR/04033/2020, UIDB/CVT/00772/2020 and by Base Funding-UIDB/00511/2020 of the Laboratory for Process Engineering, Environment, Biotechnology, and Energy LEPABE-funded by national funds through the FCT/MCTES (PIDDAC); Project 2SMART-engineered Smart materials for Smart citizens, with reference NORTE-01-0145-FEDER-000054, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). BM-F was supported by FCT, 2020.07675.BD.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "When comparing the sample by sport categories, there are statistically significant differences in somatic anxiety (p = 0.036) and deconcentration (p = 0.034), as well as in LOT-total (p \u2264 0.001) and pessimism (p \u2264 0.001). In relation to the sport modalities, lifeguards show more anxiety 38.39 (0.49) and more commitment 4.58 (0.87) while futsal athletes reach higher scores in deconcentration 8.45 (2.29). It is concluded that the variables of commitment and resilience had a statistically significant positive effect, and the category of <16 years had a statistically significant negative effect, so the lower the category, the higher the optimism.This research aims to analyse the differences in optimism, resilience, engagement and competitive anxiety as a function of the sport modality practiced in lifeguarding and futsal (team sport); the sport category by age (cadet or youth) and gender. The LOT-R optimism questionnaire, the Connor-Davidson Resilience Scale (CD-RISC-10), the Utrecht Work Engagement Scale (UWES) and the Competitive Anxiety Scale (SAS-2) were applied to a sample of 189 participants (139 men and 50 womwn) aged between 14 and 17 years. The following statistical tests are performed: Cronbach's alpha, Pearson's linear correlation, Student's t-test, Kolmogorov-Smirnov test, Levene's test and multivariate linear regression. The data indicate that there are significant gender differences in total anxiety (p <0.001) and all its dimensions , also in optimism and pessimism ( Sport research related to stress and well-being factors has been widely developed in recent years L\u00f3pez, , consideThe present study contributes to the literature on psychological elements affecting athletes by examining the competitive context in the disciplines of lifeguarding, individual sport and futsal team sport.First, the constructs, anxiety, resilience, optimism and engagement are analyzed in the sport domain.Athletes during competitions are exposed to an environment conducive to generate anxiety symptoms. Anxiety can be defined as a future emotional state characterized by a sense of apprehension, worry and lack of control over one's emotional response show lower levels of commitment and resilience than lifeguards .H4, futsal players have less anxiety than lifeguards.All of the above leads us to the objective of the study, which focuses on finding out the existing differences between lifesaving and futsal (team sport) from a psychological perspective, as well as determining differences in relation to the sporting categories, sex and age of the participants, providing answers to the different hypotheses put forward.A cross-sectional, observational and quantitative design was carried out, selecting the sample in a non-probabilistic or intentional way. The study, All the athletes were asked, through their coaches, to collaborate in the research. All the minors, in the lifeguard modality, in the course of the processing of the sports license for the year included a section in which they accepted to participate as sample subjects in possible non-invasive studies that could be carried out. Once informed consent had been obtained from the parents/guardians/legal guardians of the futsal players, all those who wished to participate in the study filled in the questionnaires prior to the celebration of a Spanish championship for the lifeguards and before a league match for the futsal players. Data were collected before the covid-19 pandemic, from the lifesaving athletes, on the day of the competition, in an after-breakfast meeting, all were concentrated in a hotel, and from the futsal players, before the pre-match talk, about 2 h before the lifesaving competition and matches. Their preparation and level of physical work was part of the peak performance phase, taking into account that all participants trained 3\u20134 h per week. The study was conducted under the guidelines of the Declaration of Helsinki, the design, observational, did not contain any ethical aspects that required prior authorization from the Bioethics Committee of the University of Murcia, Spain.The questionnaire consisted of a total of 54 items comprised of four separate and established questionnaires measuring resilience, competitive anxiety, optimism and engagement, as well as socio-demographic data. Resilience (RS) was measured using the Connor-Davidson Resilience Scale (CD-RISC-10) . Cronbach's alpha was calculated in order to check the reliability of the different scales within this particular sample, and correlations between variables were calculated using Pearson's linear correlation coefficient (r). In addition, for quantitative variables, the t-Student test was performed for the comparisons of means between two groups. Assumptions of normality and uniformity of the variances required for mean comparisons were tested using the Kolmogorov-Smirnov test and the Levene test, respectively. Finally, a multivariate linear regression model was developed to determine the possible effects of sex , sport modality and category (cadet or youth) on the resilience, anxiety and engagement scales. Statistical analyses were performed using SPSS 25.0 for Windows . Statistical significance was defined as The mean , Cronbach's alpha reliability indexes, and the correlations between the different subscales of the questionnaires used that address each variable chosen. Internal consistency rates were all higher than 0.79, indicating high reliability.In As for the LOT-R scale, the LOT-T correlated positively and statistically significantly with the LOT-O, UWES-T, UWES-V, UWES-D, UWES-A and RS, however, it correlated negatively and statistically significantly with the LOT-P. The LOT-O dimension was positively and statistically significantly correlated with LOT-P and SR and negatively and statistically significantly correlated with UWES-A. The LOT-P dimension had a negative and statistically significant relationship with the UWES-T, UWES-D, UWES-A and resilience.The UWES-T showed positive and statistically significant differences with the UWES-V, UWES-D, UWES-A and SR. The UWES-V dimension showed positive and statistically significant differences with UWES-D, UWES-A and RS, and the UWES-D dimension showed positive and statistically significant differences with UWES-A and RS.Focusing on the differences between males and females in the sample, Regarding the sport category variable, higher A-S was found in the juvenile group (<18 years) and higher A-D in the cadets (<16 years), as shown in When the sample was divided according to sport modality , there were statistically significant differences between the groups in A-T, and its dimensions, A-S, A-C and A-D, F = 14.1; p < 0.001, explaining 31.7% of the explanatory variance. The variables of commitment and resilience had a significant positive effect, and the category of <16 years had a significant negative effect, so the lower the category, the higher the optimism.To determine in LOT-R the effects of the variables sex, type of sport and sport category (<18 and <16 years) and anxiety, UWES and RS, a linear regression was performed. The results are shown in In sport competition, it is essential to know and control the factors that influence performance and futsal (team sport) modalities; from a psychological perspective, as well as determining differences in relation to the sport categories, sex and age of the participants, responding to the different hypotheses put forward.On analyzing the correlations of the different scales used in the study, we found a relationship between the level of LOT-T and SR, as well as between the UWES-T and SR. Likewise, we found a negative correlation between the level of A-D and SR, data that support previous studies Trigueros et al. and RechThe present data reflected a higher level of A-T and UWES-T in boys, as well as lower levels of A-D and higher levels of LOT-O in girls. Thus, partly corroborating H1, boys have lower A-T, UWES-T and RS than girls. This finding is in line with some previous studies and cadets (<16 years), the data show a similar result to previous studies, such as those of Vallarino and Reche-Garc\u00eda , who fouWhen correlating the different scales in terms of sport modality, in A-T, LOT-T, UWES-T and RS, it is the lifeguard athletes who show higher scores than the futsal athletes. Which determines that H3, futsal athletes (collective sport) show a lower level of commitment and resilience than lifeguards , H1 is confirmed. Regarding the sport modality and the relationship between collective sport and individual sport (lifeguard), we found similar results to previous studies, such as Boh\u00f3rquez and Checa and RechWe believe that the results presented are due to the specific characteristics of the sports studied. On the one hand, lifeguarding is a sport in which the work is carried out individually but within a team discipline, which we believe reduces the possibility of differences between its practitioners. Whereas, in Indoor Football, the possibility of distributing responsibility among the team, we understand, is the reason for these differences between modalities. As determined by Vallarino and Reche-Garc\u00eda who highIn relation to H4, Futsal players have lower A-T than lifeguards, H3 is endorsed. Since the lifeguard athletes achieved higher scores than the futsal players. Possibly the cause could be as a consequence of the sport modality practiced as when one realizes that one is unable to achieve a goal it is probably very stressful, as stress occurs when goals become unattainable Lazarus, . ConsideThe results of the linear regression revealed the importance of the category, the level of UWES-T and the level of SR of the participants. Unlike previous studies, S\u00e1nchez-Oliva et al. or Chac\u00f3The results of this study provided very valuable information about optimism, anxiety, resilience and the level of commitment of athletes <18 years old and <16 years old, both in lifeguarding and futsal. From the data of this study, it will be possible to establish a better working environment in training and provide psychological work situations so that athletes can obtain greater positive feelings, both in individual and collective sport. Providing coaches and athletes with information on the dimensions analyzed, in order to be able firstly to detect if there are risks in athletes and secondly to prevent them from appearing, with the inclusion in training and/or training of athletes of efficient tools and methodologies in the detection, prevention and reinforcement of concepts such as LOT-T and RS that help athletes and coaches to face them with solvency. Using the present research as a reference, additional future research is needed in other sports, both individual and team, before stronger conclusions can be drawn. Larger sample sizes, longitudinal studies, including pre- and post-competition observation may provide more information on the more concrete effects of these variables on athletes. Such research may also reveal whether the results of the questionnaires are different depending on the athlete's performance in an event.It is worth noting that higher anxiety and UWES ability was observed in the lifeguarding modality, and higher levels of deconcentration and LOT-R-P were observed in futsal participants.The study provides insight into the athletes of these two sport modalities on which to deepen future research, generate personalized lines of work and improve performance. We are also aware of the need to continue increasing the sample for future replications, and even generate sport-specific studies and comparing different competitive levels.With regard to the limitations of the study, firstly, we can point out that the data have been collected through self-reporting. This is a common practice in studies, although it may lead to a bias in the participants' response, exacerbate common variance and artificially increase correlations between variables Spector, . SecondlThe raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Ethical approval was not provided for this study on human participants because as it was an observational study and the information was obtained through a questionnaire, in which there was no sensitive information. The ethics committee determined that there was no need for such a report. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.FC-N and AM-M: conceptualization, writing\u2014review and editing, and supervision. FC-N: methodology, writing\u2014original draft preparation, and project administration. FC-G and RI-P: software and visualization. AM-M, RI-P, and FC-N: validation. AM-M: formal analysis. FC-N and RI-P: investigation. FC-G and AM-M: resources. RI-P: data curation. AM-M, RI-P, FC-G, and FC-N: funding acquisition. All authors have read and agreed to the published version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Ankylosing spondylitis (AS) is a chronic inflammatory disease. Excess cardiovascular risks were well recognized in patients with AS and were attributed to prolonged systemic inflammation. Uveitis is one of the most common extra-articular symptoms of AS and is also considered an indicator of systemic inflammation. This study aimed to investigate whether uveitis was a risk factor for developing acute myocardial infarction (AMI) in patients with AS using the National Health Insurance Research Database (NHIRD).Data were collected from the NHIRD over a fifteen-year period. Variables were analyzed using the Pearson chi-square test and Fisher\u2019s exact test. Risk factors for the occurrence of AMI were examined by calculating hazard ratio. Kaplan-Meier analysis was performed to compare the cumulative incidence of AMI in the uveitis and non-uveitis cohorts.p < 0.001). Stratified analysis revealed that patients with uveitis had an increased risk of developing AMI regardless of their sex . Patients with uveitis in all age groups were independently associated with an increased risk of developing AMI compared to those without uveitis . Patients with uveitis had a higher probability of developing AMI regardless of comorbidities. Uveitis patients with comorbidities had a higher risk of developing AMI compared to uveitis patients without comorbidities.A total of 5905 patients with AS were enrolled, including 1181 patients with uveitis (20%) and 4724 patients without uveitis (80%). The Kaplan\u2013Meier method with the log-rank test showed that the uveitis group had a significantly higher cumulative hazard for patients with AMI than the non-uveitis group (p < 0.001). The adjusted hazard ratio (aHR) of AMI was higher in the uveitis group than in the non-uveitis group (aHR = 1.653, Uveitis is a significant risk factor for developing AMI in patients with AS. Physicians should be aware of the potential cardiovascular risk in AS patients with uveitis, especially simultaneously with other traditional risk factors of AMI. Further prospective studies are needed to elucidate the underlying mechanism between uveitis and AMI in patients with AS. Ankylosis spondylitis (AS) is a chronic inflammatory disease that predominantly affects the spine, with a peak onset between the ages of 20 years and 40 years . More thUveitis is one of the most common extra-articular symptoms of AS. The prevalence of uveitis in AS is approximately 20\u201330% . CurrentAs AMI is associated with life-threatening consequences , it is important to know how AS and AMI are associated, including the risk factors of AMI in AS. However, most studies on this subject are case reports or case series, and the lack of a large sample size to investigate the association. Therefore, we used the National Health Insurance Research Database (NHIRD), which contains a comprehensive longitudinal medical record from 23 million people in Taiwan. This approach resulted in a large number and varying features of patients with AS compared to previous studies. This study aimed to investigate the factors that determine the incidence of AMI in AS. In addition, the study elucidated whether uveitis could be a risk factor for the development of AMI in AS. Our ultimate goal is to raise the awareness of physicians about AMI in AS patients with uveitis and provide information to all physicians for multidisciplinary care.Beginning in 1995, the National Health Insurance (NHI) program was launched in Taiwan. More than 23 million Taiwanese and non-Taiwanese residents were included. The National Health Insurance Research Database (NHIRD) contains comprehensive medical records, including outpatients, inpatients, diagnoses, prescriptions, and procedures, which are registered by the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) codes. The ICD-9-CM codes used in this study are listed in We conducted a retrospective cohort study between January 2000 and December 2015. All included patients had at least one inpatient claim or outpatient visits more than three times with a diagnosis of AS according to the ICD-9-CM code . Patients younger than 20 years old, with unknown sex, or without tracking were excluded. Patients diagnosed with AS and AMI before the index date were also excluded to ensure that patients had a new onset of AS and new onset of AMI after AS diagnosis. The index date was defined as the date of the first AS diagnosis. The study population was divided into the uveitis group and non-uveitic group (n = 18815). The group of non-uveitic patients matched four-fold with the uveitic group for sex, age, comorbidities, and index date as the comparison cohort (n = 4724). The uveitis group was divided into the anterior uveitis and posterior segment involvement groups. The populations in both groups did not overlap with each other. The uveitic subgroups and comorbidities extracted by ICD-9-CM codes are listed in The study protocol was reviewed and approved by the institutional review board of the Tri-Service General Hospital (TSGH IRB No. B-110-35). Because the identities of all patients in the NHIRD were encrypted before data were released, the requirement for signed informed consent of the included patients was waived.The Pearson chi-square test and Fisher\u2019s exact test were used to evaluate differences in categorical variables such as sex, age group, and comorbidities. The threshold for statistical significance was set at p < 0.05. A Cox proportional hazard model was used to estimate the hazard ratio (HR) for the occurrence of AMI based on each variable in the univariate and multivariate analyses. Considering competing risk, Cox regression with Fine and Gray\u2019s competing risk model was also employed for analysis. Kaplan\u2013Meier analysis was performed to estimate the cumulative incidence of AMI in these two cohorts. All statistical analyses were performed using SPSS software .The cumulative risk of developing AMI in patients with AS was calculated using the Kaplan\u2013Meier method was further divided into the anterior uveitis group and posterior segment involvement group compared to the non-uveitis group (n = 4724) for developing AMI than patients without uveitis. Kaplan\u2013Meier analysis also revealed that the cumulative risk of developing AMI was significantly increased in the uveitis group. In the stratified analysis, the incidence of AMI in the uveitis group was 1306.54 per 100,000 person-years and 1.47-fold higher than that in the non-uveitis group. Uveitis was associated with an increased risk of developing AMI in both sexes, all age groups, patients with and without comorbid DM, hyperlipidemia, HTN, CVA, CHF, COPD, asthma, CAD, and metabolic syndrome. Uveitis patients with comorbidities had a higher adjusted hazard of developing AMI compared to uveitis patients without comorbidities.The prevalence of AS has geographical differences. AS is more common in Europe and Asia than in Latin America . PreviouFor comorbidities, the percentages of hypertension and chronic obstructive pulmonary disease were similar to those reported in previous studies , 26. TheOur report also identified uveitis as an independent risk factor for AMI in patients with AS , 31, 32.Previous studies have reported that uveitis in spondyloarthritis is associated with HLA-B27, possibly acting through the IL-23/IL-17 axis \u201338. The We further divided uveitic patients into the anterior uveitic and posterior segment involvement groups \u201347. ThisOur study has several strengths. First, the National Health Administration in Taiwan checks charts to ensure patient diagnosis with appropriate treatment. Treatment followed a standardized protocol. The diagnoses were verified. Second, we used a comprehensive, longitudinal, nationwide database spanning 15 years. The NHI system was introduced in Taiwan in 1995, so we could conduct a longitudinal analysis of sequential events. AS or AMI before the index date could be excluded to eliminate bias in the cross-sectional study. Third, the coverage rate in Taiwan was approximately 99%. A large sample provides high statistical power. Finally, the major strength was that the findings after adjusting for confounders were highly consistent, suggesting reliable and convincing results.However, this study had some limitations. First, the study was based on a retrospective statistical analysis from a database. The NHIRD database does not provide laboratory data, such as the erythrocyte sedimentation rate or C-reactive protein, as advocatory evidence. There were no clinical images such as MRI findings for classification of disease activity and severity of AS patients, which might also show a correlation with uveitis. Second, some traditional factors for AMI, such as diet, smoking, and physical activity, were not in the database. Third, AS-uveitis complex is most frequently observed as anterior uveitis and responds well to topical corticosteroids, whereas posterior uveitis is usually refractory to topical treatments and frequently requires treatments with systemic corticosteroids and anti-TNF agents to reduce recurrences. Cardiovascular events are complications of corticosteroids. In this study, we did not assess the influence of uveitis treatments in developing AMI. Fourth, the rates of all extra-articular manifestation in AS patients might influence the development of AMI, but the database did not provide the information of all extra-articular manifestation. Fifth, the included patients in our study are homogenous ethnic population with specific genetic and disease features as indicated by the prevalence of co-morbidities, type of uveitis and probably age at diagnosis. Hence, the results of our study must be confirmed in other ethnic populations. Sixth, this study had a retrospective design. We could only elucidate a possible mechanism based on this association. The causality needs further prospective studies with detailed information to confirm our conclusion.Uveitis is a risk factor for developing AMI in patients with AS. Doctors should be aware of the potential cardiovascular risk in patients with AS uveitis, especially simultaneously with other traditional risk factors for AMI. Closely monitoring the cardiovascular system in these patients is important for all physicians to detect AMI earlier and manage it promptly. Further prospective studies are needed to elucidate the underlying mechanism between uveitis and AMI in patients with AS.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by The institutional review board of the Tri-Service General Hospital (TSGH IRB No. B-110-35). Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.Conceptualization: Y-FL, W-CC, and C-LC. Methodology: W-CC and C-HC. Software: W-CC, C-AS, and C-HC. Validation: T-YL, W-CC, and C-HC. Formal analysis: W-CC, C-AS, and C-HC. Investigation: T-YL, Y-HC, and J-TC. Resources: W-CC, Y-HC, J-TC, and C-LC. Data curation: W-CC, C-HC, and C-LC. Writing\u2014original draft preparation: Y-FL. Writing\u2014review and editing: Y-FL, T-YL, and C-LC. Visualization: W-CC and C-HC. Supervision: W-CC, Y-HC, and C-LC. Project administration, Y-HC and C-LC. Funding acquisition, W-CC, J-TC, and C-LC. All authors have read and agreed to the published version of the manuscript.This study was supported by the Tri-Service General Hospital Research Foundation and by the Ministry of National Defense . The sponsor has no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "We aimed to develop and validate a new nomogram for predicting the risk of intracranial hemorrhage (ICH) in patients with acute ischemic stroke (AIS) after intravenous thrombolysis (IVT).n = 387) and the testing set . The factors in the predictive nomogram were filtered using multivariable logistic regression analysis. The performance of the nomogram was assessed based on the area under the receiver operating characteristic curve (AUC-ROC), calibration plots, and decision curve analysis (DCA).A retrospective study enrolled 553 patients with AIS treated with IVT. The patients were randomly divided into two cohorts: the training set . The calibration plot demonstrated good agreement in both the training and testing sets. DCA indicated that the nomogram was clinically useful.After multivariable logistic regression analysis, certain factors, such as smoking, National Institutes of Health of Stroke Scale (NIHSS) score, blood urea nitrogen-to-creatinine ratio (BUN/Cr), and neutrophil-to-lymphocyte ratio (NLR), were found to be independent predictors of ICH and were used to construct a nomogram. The AUC-ROC values of the nomogram were 0.887 (95% CI: 0.842\u20130.933) and 0.776 (95% CI: 0.681\u20130.872) in the training and testing sets, respectively. The AUC-ROC of the nomogram was higher than that of the Multicenter Stroke Survey (MSS), Glucose, Race, Age, Sex, Systolic blood Pressure, and Severity of stroke (GRASPS), and stroke prognostication using age and NIH Stroke Scale-100 positive index (SPAN-100) scores for predicting ICH in both the training and testing sets (The new nomogram, which included smoking, NIHSS, BUN/Cr, and NLR as variables, had the potential for predicting the risk of ICH in patients with AIS after IVT. Stroke is the second leading cause of death and a major leading contributor to disability worldwide . IntraveSince ICH may lead to poor prognosis, a reliable scoring tool that can be used to identify the risk of post-thrombolysis ICH is essential. Several scoring systems have been applied to predict the risk of ICH after thrombolysis \u201310. HoweIn this study, we aimed to develop a new nomogram to predict the risk of ICH after IVT therapy and to compare the performance of our model with that of other scoring systems for predicting ICH. The nomogram we developed will help clinicians identify patients with AIS with a higher risk of ICH after IVT therapy.The present study was a retrospective study performed in the stroke center of the Central Hospital of Shaoyang between November 2013 and January 2021. The patients met the following inclusion criteria: 1) more than 18 years of age; 2) diagnosed with AIS confirmed by MRI within 24 h after admission; 3) onset-to-treatment time for thrombolysis of <4.5 h; 4) they or their legal representatives provided written informed consent to participation in the study; and 5) treated with IVT. The exclusion criteria were as follows: 1) treated with endovascular procedures after IVT; and) not treated with IVT or refused IVT treatment; and 3) medical contraindications for IVT.This study was approved by the Ethics Committee of the Central Hospital of Shaoyang. The Ethics Committee approval letter is available in Online Resource 1.We collected demographic, clinical, and laboratory information at admission and during hospitalization. The baseline data included age, sex, current smoking status, current drinking status, NIHSS score on admission, systolic blood pressure (SBP), diastolic blood pressure (DBP), blood glucose level on admission, history of hypertension and diabetes mellitus, atrial fibrillation (AF), international normalized ratio (INR), activated partial thromboplastin time (APTT), PLT, fibrinogen, albumin (ALB), white blood cells (WBC), neutrophil-to-lymphocyte ratio (NLR), high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglyceride (TG), total cholesterol (TC), blood urea nitrogen-to-creatinine ratio (BUN/Cr), and uric acid (UA).All patients received rt-PA treatment within 4.5 h after the onset of stroke. Intravenous rt-PA at 0.9 mg/kg (90 mg maximum) was used; 10% of the total dose was administered as an intravenous bolus, followed by infusion of the remaining dose over 60 min .All patients underwent a CT scan on admission, and each patient underwent another CT scan 24 h after intravenous rt-PA administration. In cases in which the patient's neurological function deteriorated rapidly, a CT scan was immediately performed to assess the presence of ICH. According to the European Cooperative Acute Stroke Study-2 criteria , the disn = 387) and the testing set (n = 166), at a theoretical ratio of 7:3. Categorical variables and continuous variables are expressed as frequencies and means (SDs) or medians , respectively. The differences in baseline characteristics between the training set and the testing set were assessed using Student's t-test or the non-parametric Mann\u2013Whitney U test for continuous variables and the \u03c72 or Fisher's exact test for categorical variables. The least absolute shrinkage and selection operator (LASSO) method was used to screen the optimal risk factors related to ICH in the total dataset. The variables with non-zero coefficients in the LASSO regression model were selected for further analysis. In the training cohort, univariable logistic regression analysis was used to predict the probability of ICH. The variables with P-value <0.05 were added to the multivariable logistic regression analysis to screen for independent clinical predictors related to ICH. A nomogram was generated based on these risk factors in multivariable analysis. The area under the receiver operating characteristic curve (AUC-ROC) analysis was performed to assess the predictive accuracy of the nomogram, and calibration curves were drawn to compare the predicted probabilities with the observed probabilities. Decision curve analysis (DCA) was used to evaluate the clinical value of the predictive model.A total of 553 patients were randomly divided into two groups, the training set score, the Glucose, Race, Age, Sex, Systolic blood Pressure, and Severity of stroke (GRASPS) score, and the stroke prognostication using age and NIH Stroke Scale-100 positive index (SPAN-100) score. The AUC-ROC was used to estimate the accuracy and discrimination of the nomogram and these scoring systems in the training and testing sets. DeLong's test was performed to compare the differences in AUC-ROC between the ICH nomogram and the MSS score, GRASPS score, and SPAN-100 score.http://www.R-project.org/) and SPSS version 27.0 . Two-tailed values of p < 0.05 were considered statistically significant.Statistical analyses were performed using R version 3.6.3 software .The training set consisted of 387 individuals, and the remaining 166 individuals were included in the testing set. The baseline characteristics of the patients in the training and testing sets are shown in Through searching and analysis of the related literature, 24 potential risk factors among the demographic, clinical, and laboratory indicators of the patients were included in the LASSO regression analysis . The varp < 0.05), as shown in A univariable analysis of the training set revealed that smoking, AF, NIHSS, BUN/Cr, and NLR were related to ICH. These factors were therefore used in multivariable logistic regression analysis for screening independent clinical predictors of ICH. Multivariable logistic regression analysis demonstrated that four variables are independently associated with ICH were used to build a nomogram for predicting the risk of ICH in AIS patients with IVT . The ratp < 0.05). Similar to the results obtained with the training set, the value of AUC-ROC in our model was superior to the values obtained using the MSS scores , the GRASPS scores , and the SPAN-100 scores in the testing set (p < 0.05).To assess the performance of the nomogram in predicting ICH, the likelihood that each patient would experience ICH was also predicted based on his or her MSS scores, GRASPS scores, and SPAN-100 scores. We compared the discrimination of our nomogram with those obtained using MSS scores, GRASPS scores, and SPAN-100 scores using AUC-ROC. As shown in We developed a new nomogram based upon smoking, NIHSS scores, BUN/Cr, and NLR for use in predicting the risk of ICH after IVT therapy. The developed nomogram demonstrated good discrimination and calibration in the training and testing sets. Furthermore, the AUC-ROC curve obtained using our nomogram showed better performance than the curves obtained using the MSS scores, the GRASPS scores, and the SPAN-100 score in both the training and testing sets. In addition, the DCA results suggested that the developed nomogram has a marked net benefit for predicting the risk of ICH.p < 0.05). The better performance of our nomogram may be explained as follows. First, the three scoring systems with which it was compared converted continuous variables to dichotomization/categorization variables; this conversion is statistically inefficient and may decrease the accuracy of prediction scores and the hemorrhage after thrombolysis (HAT) scores rely on CT images to detect early ischemic changes and hyperdense cerebral artery signs , 7. Howeediction . Second,ediction , 22. In ediction \u201325.n = 345). In addition, the percentage of ICH based on this nomogram was 14.5%, significantly higher than the percentage of ICH in Asian patients nomogram was constructed to predict ICH in stroke patients after IVT treatment in Italy . Howeverpatients , 30. TheIn this study, smoking, NIHSS scores, BUN/Cr, and NLR were shown to be independent predictors of ICH in patients with AIS after rt-PA administration. Consistent with previous studies, smoking and NIHSS were common risk factors for predicting ICH. A previous study reported that age \u226568 years, smoking, AF, SP \u2265149 mmHg 2 h after rt-PA administration, and NIHSS scores \u226517 before thrombolysis were associated with the risk of ICH after IVT . In WangThe NLR is another new risk factor we found that can contribute to the risk of ICH. Neuroinflammation is related to the entire pathological process of stroke. The NLR is a readily accessible and reproduces new inflammatory biomarker . It was Generally, the nomogram developed in this study, which includes four parameters, is convenient and efficient for use in the management of patients with AIS. First, the predictors used in our predictive nomogram are easily available at almost all medical centers, even those in poor areas. In addition, the nomogram is particularly suitable for use by non-neurologists because CT imaging is excluded. Furthermore, the discrimination and calibration performance of the nomogram was good; therefore, it could be a reliable tool for predicting the risk of ICH in patients with AIS after rt-PA treatment.Our study has some limitations. First, our data came from a single-center retrospective analysis, and this might have limited the statistical power of the results. Second, our model has not been validated in external cohorts. Prospective, multicenter studies will be required in the future to assess the applicability of our nomogram. Third, data on onset-to-treatment time for thrombolysis or administration of oral antiplatelet drugs or anticoagulants, all of which may be risk factors for ICH, were not available in our study.In conclusion, the new nomogram that includes smoking, NIHSS, BUN/Cr, and NLR may predict the risk of ICH after IVT in AIS patients in the Asian population.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Ethics Committee of the Central Hospital of Shaoyang. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.A-DX, DL, Z-AW, and X-XH contributed to the study conception and design. Material preparation, data collection were performed by DD, Z-GY, S-YL, J-KZ, Y-FLi, Y-FLiu, Y-SW, T-YZ, and X-LS. Analysis and interpretation of the data were done by Z-AW. The first draft of the manuscript was written by Z-AW, X-XH, A-DX, and DL. All authors read and approved the final manuscript.This work was supported by grants from the National Natural Science Foundation of China , the Science and Technology Planning Project of Guangdong Province, China (2017A020215049 and 2019A050513005), Natural Science Foundation of Guangdong Province (2018A0303130182 and 2020A1515010279), Postdoctoral Research Foundation of China (2018M643370 and 2021M701419), Guangzhou Science and Technology Planning Project (201508020004), Technology and People's Livelihood Major Project of Guangzhou (2014Y2-00505), and the Fundamental Research Funds for the Central Universities (21621102).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "In \u201cScreening Tools: Their Intended Audiences and Purposes. Comment on \u2018Diagnostic Accuracy of Web-Based COVID-19 Symptom Checkers: Comparison Study\u2019\u201d :e26148), one error was noted.In the originally published manuscript, an incorrect statement was included in the Conflicts of Interest section:EM, MF, and SG are employees of Ada Health GmbH. AG has no conflicts to declare.The statement has been corrected as follows:All authors are employees of Ada Health GmbH.The correction will appear in the online version of the paper on the JMIR Publications website on June 24, 2021, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "The differences were statistically investigated, and the features were classified using Support Vector Machine (SVM). Results: The results show that medication has a significant effect on the change of time and amplitude perturbation (jitter and shimmer) and harmonics of /m/, which was the most sensitive individual phoneme to the levodopa response. /m/ and /o/ performed at a comparable level in discriminating PD-off from control recordings. However, SVM classifications based on the combined use of the three phonemes /a/, /o/, and /m/ showed the best classifications, both for medication effect and for separating PD from control voice. The SVM classification for PD-off versus PD-on achieved an AUC of 0.81. Conclusion: Studies of phonation by computerized voice analysis in PD should employ recordings of multiple phonemes. Our findings are potentially relevant in research to identify early parkinsonian dysarthria, and to tele-monitoring of the levodopa response in patients with established PD.Background: Parkinson\u2019s disease (PD) is a multi-symptom neurodegenerative disease generally managed with medications, of which levodopa is the most effective. Determining the dosage of levodopa requires regular meetings where motor function can be observed. Speech impairment is an early symptom in PD and has been proposed for early detection and monitoring of the disease. However, findings from previous research on the effect of levodopa on speech have not shown a consistent picture. Method: This study has investigated the effect of medication on PD patients for three sustained phonemes; /a/, /o/, and /m/, which were recorded from 24 PD patients during medication I.Parkinson\u2019s disease (PD) is the second most common neurodegenerative disorder The Movement Disorder Society Unified Parkinson\u2019s Disease Rating Scale Part III (MDS-UPDRS-III) One of the early symptoms of PD is change in voice, which can precede other motor features Studies on the voice or speech parameters can be divided into four groups based on the analyzed aspect: phonatory, articulatory, prosodic, and linguistic et al.et al.et al.et al.Numbers of studies have investigated the voice parameters obtained from sustained phonemes to determine the differences between PD patients and healthy participants The voice features that have shown a significant difference between the voice of healthy and PD patients are pitch frequency, jitter, shimmer, and harmonics to noise ratio et al.et al.et al.et al.et al.The motor symptoms of PD are managed by dopaminergic pharmacological treatments, of which levodopa is the most effective and widely used. Most patients improve on levodopa, though one weakness of the drug is the tendency for an unstable, fluctuating response to develop after a number of years et al.et al.et al.et al.et al.et al.et al.Studies that have primarily focused on phonation in PD do not show a consensus about levodopa effect. Sanabria The aim of this study was to investigate the use of phonatory parameters to classify PD patients before and after levodopa medication. We examined the change in phonatory parameters by using a statistical hypothesis test and the Support Vector Machine (SVM) algorithm to separate the two PD medication states and the control groups. Three different sustained phonemes were considered: /a/, /o/ and /m/. These phonemes were selected to examine a range of voice production II.A.Twenty-four PD patients were recruited from the Movement Disorders Clinic at Monash Medical Centre. All had been diagnosed within the last ten years and complied with the Queen Square Brain Bank criteria for idiopathic PD off state (PD-off) . They were retested in the on state (PD-on), taken to be the maximum improvement 30 \u2013 90 minutes after a subject\u2019s usual morning levodopa dose. Motor function in off and on states was scored by a neurologist on the MDS-UPDRS-III PD participants were first assessed in a practically defined The study protocol was approved by the ethics committee of Monash Health, Melbourne, Australia (LNR/16/MonH/319) and RMIT University Human Research Ethics Committee, Melbourne, Australia (BSEHAPP22-15KUMAR). Before the experiments, written informed consent was obtained from all the participants.B.Three sustained phonemes /a/, /o/, and /m/ were recorded from each participant. The participants were instructed to pronounce the vowel for as long as it was comfortable, with their natural pitch and loudness.The phonemes were recorded using Samson-SE50, an omnidirectional head-worn microphone. The recordings were saved into a single-channel uncompressed WAV format with a sampling rate of 48 kHz and a 16-bit resolution. Each recording contained one single sustained phoneme of 5.1 to 38.6 seconds duration as shown in C.MATLAB2018b (MathWorks) was used for all analyses. Each recording was manually segmented to eliminate any unwanted sections such as silent pieces and the voice of the instructor. Based on the assumption that vowels correspond to largely stationary signals, and the need for more samples for the purpose of cross-validation, each recording was divided into ten segments of 0.5 seconds each, and the jitter, shimmer, pitch, and harmonics parameters of each segment was calculated. Each segment was considered as an individual example.The features of each segment were calculated using Praat abs), jitter relative (rel), relative average perturbation (rap), and period perturbation quotient-5 (ppq5). The rap and ppq5 are the perturbation of the difference between Four jitter parameters were extracted from the recordings: jitter absolute , the relative shimmer, HNR and NHR were calculated based on the normalized autocorrelation function of the segment. HNR and NHR were calculated as described in The pitch parameters are the mean, median, standard deviation, maximum, and minimum of the instantaneous pitch frequency D.on and PD-off to determine the effect of medication on the patient. Mann Whitney U-test off, and CO and PD-on.All the statistical analyses were performed using MATLAB2018b (MathWorks). The normality of the extracted parameters was examined with the Anderson-Darling test off and PD-on MDS-UPDRS-III scores.The The 95% confidence level was considered for the analysis and E.Support Vector Machines (SVM) The size of dataset for CO subject was For each SVM training, only 80% of the training sets (randomly picked) were used, while the other 20% were used for testing , true-negative (TN), false-positive (FP), and false-negative (FN). The Receiver Operating Characteristic (ROC) curve was generated, and the Area Under Curve (AUC) was calculated for each SVM model.III.A.on and PD-off.Anderson-Darling test confirmed that the voice parameters for the three groups and the three phonemes were not normally distributed and thus unsuitable for parametric test. Mann Whitney U test was used to test for group differences in each of the features. Wilcoxon signed-rank test was used to test the differences between dependent data of PD-1)on and PD-off, and this identifies the features that are significantly changed by medication. It shows that p-value was less than 0.05 for most of the parameters of phoneme /m/. The jitter and harmonics parameters of phoneme /o/, as well as the pitch of phoneme /m/ were also changed due to medication.2)off are demonstrated in The Mann Whitney U-test results for group differences between CO and PD-3)on. The results show that jitter, shimmer, and harmonics parameters of phoneme /o/ of the two groups were well separated. The jitter of phoneme /a/ and /m/ were effective to differentiate CO and PD-on.4)on and PD-off. Phoneme /m/ was effective to identify the change due to medication as well as differentiating between CO and PD-off. It is also seen that harmonics parameters of /o/ and /m/ were statistically different between PD-off and PD-on.Comparison of the results on the above statistical analysis, confirm that the majority of the parameters of the phoneme /o/ were significantly different between the healthy subjects (CO) and PD patients; both, PD-B.on was 0.81. Classification between CO and PD-off shows that the best result was with the combination of the three phonemes, with AUC = 0.90. The best classification result between CO and PD-on was with the combination of phonemes /a/ and /m/ with AUC = 0.86.The results of classification by SVM of the voice features of each of the three phonemes are shown in IV.off from PD-on. Time and amplitude perturbations (jitter and shimmer) decrease with medication, suggesting that levodopa improves voice quality. In differentiating control from PD-off recordings, in effect detecting parkinsonian dysarthria, all /a/, /o/ and /m/ achieved comparably good levels of statistical significance and harmonic to noise ratio of the phoneme /m/ which is confirmed also by SVM classification. Thus, /m/ can be used to differentiate between PD-The limitation of this study is that because of the relatively small number of participants, it was not possible to differentiate between genders. Further, the differences such as accents, demographics, and language skills have not been investigated. Another shortcoming of this study is that each participant was only studied once and hence the repeatability has not been checked. Recruitment of controls was subject to some conditions from Research Ethics approval to advertising, and the mean control group age is about 5 years younger than the PD patients.V.on and PD-off, using SVM, was when all three phonemes were used. While /o/ showed the greatest differences between PD and controls, the best classifications when using SVM were again obtained from combined analysis of all three phonemes. Whether attempting to separate parkinsonian dysarthria from control voice, or to detect the levodopa effect on voice in PD, this study shows that computerized analysis of multiple phonemes should be employed. Our findings are potentially relevant in research to identify early parkinsonian dysarthria, and for tele-monitoring of the levodopa response in patients with established PD.This study has investigated the effect of levodopa medication on the voice of PD patients based on utterance of three phonemes, /a/, /o/ and /m/. It has found that medication has a significant effect on the change of time and amplitude perturbation of the phoneme /m/. But the highest accuracy in differentiating PD-"} +{"text": "Paediatric inflammatory bowel disease (p-IBD) is a chronic and relapsing gastrointestinal disorder of childhood with associated long-term morbidity. Several extraintestinal manifestations (EIMs) are described, the most common being joint pain and/or inflammation. In 1986, Passo et al. were the first to describe the association of arthritis in p-IBD patients. However, since then, data on the epidemiology, patient and disease factors associated with, treatments for, and outcomes of p-IBD associated musculoskeletal (MSK) disease are not well-established.Our study aims to summarize the literature on the epidemiology of MSK EIMs in p-IBD in the era of biologics.A systematic review of the literature was performed. PubMed, Embase, Cochrane Library, Web of Science Core Collection, and CINAHL databases were searched with relevant keywords. Studies in English published from January 1, 2000 to December 21, 2020 were included. In total, 3893 papers were identified and screening was performed by two independent reviewers . Conflicts were resolved by a third reviewer . Study and population characteristics were recorded. The primary outcomes of interest were MSK symptoms at presentation and their course, method of diagnosis and definitions used for MSK EIMs. Risk of bias assessment was performed using the JBI Critical Appraisal Tools.Thirteen studies were included for full review, which were primarily single-centre observational studies with retrospective or cross-sectional design. The method of diagnosis for MSK EIMs varied across the studies, with only 4 studies stating the involvement of a rheumatologist in diagnosis. The definitions also varied, with MSK EIMs such as peripheral arthritis, axial arthritis, enthesopathy, and arthralgia included. Only 7 studies focused on MSK EIMs as their primary outcome, while the remainder reported on all p-IBD associated EIMs. There was a wide range in the prevalence of MSK EIMs from 2\u201335% . Four studies reported on the therapeutic response of MSK EIMs, and only 3 of those reported on biologic use. Risk of bias demonstrated heterogeneity in the quality of included studies.This is the first systematic review of the literature for MSK EIMs in p-IBD. Analysis was limited due to variability in study design and data-reporting methods. Included studies reported prevalence of MSK EIMs, but the ascertainment of MSK EIMs, both method and definition varied with a clear lack in standardization. Our study demonstrates the need for further research of the MSK associations of p-IBD and to explore optimal management to advance care for this group of children.None"} +{"text": "Correction to: Microbiome 10, 8 (2022)https://doi.org/10.1186/s40168-021-01202-xFollowing publication of the original article , the autFrom: \"JP, ML, and ZL designed the experiment; WX collected the samples, with the contribution of ZS and CD; JP and ZC performed the informatics work; JP, LL, and ML wrote the paper, with the contributions from all authors.\"To: \"JP, ML, and ZL designed the experiment; ZS and CD collected the samples; WX performed the laboratory works; JP and ZC performed the informatics works; JP, LL, and ML wrote the paper, with the contributions from all authors.\"The authors apologize for this error."} +{"text": "Near-infrared spectroscopy (NIRS) is a fast and powerful analytical tool in the food industry. As an advanced chemometrics tool, multi-way analysis shows great potential for solving a wide range of food problems and analyzing complex spectroscopic data. This paper describes the representative multi-way models which were used for analyzing NIRS data, as well as the advances, advantages and limitations of different multi-way models. The applications of multi-way analysis in NIRS for the food industry in terms of food process control, quality evaluation and fraud, identification and classification, prediction and quantification, and image analysis are also reviewed. It is evident from this report that multi-way analysis is presently an attractive tool for modeling complex NIRS data in the food industry while its full potential is far from reached. The combination of multi-way analysis with NIRS will be a promising practice for turning food data information into operational knowledge, conducting reliable food analyses and improving our understanding about food systems and food processes. To the best of our knowledge, this is the first paper that systematically reports the advances on models and applications of multi-way analysis in NIRS for the food industry. Near-infrared spectroscopy (NIRS) is a fast and powerful analytical tool. It has gained widespread acceptance in both the scientific community and industry. NIRS uses the near-infrared region of the spectrum, commonly defined from 780 nm to 2500 nm, where most absorption bands are molecular overtones or combination vibrations bands . CompareThe rapid development of chemometrics during the past decades is progressing and advancing a lot of scientific areas which are highly overlapped with analytical science, including food science . CombineIn many scientific fields, a multi-way data analysis is popular and frequently appears under the name tensor analysis . The mulIn this paper, we focus on models and applications of multi-way analysis combined with NIRS for both food products and food process analyses. The contents of this paper are organized as follows. The second part explains the used notations and abbreviations. The third part describes the multi-way models which were used for analyzing NIRS data, as well as the advances, advantages and limitations of different models. The preprocessing techniques for multi-way analysis for analyzing NIRS data are briefly described in the fourth part. In the fifth part, the applications of multi-way analysis in NIRS for the food industry are reviewed in terms of food process control, quality evaluation and fraud, identification and classification, prediction and quantification, and image analysis. The available software and algorithms for multi-way analysis are introduced in the sixth part. The conclusions are formulated in the last part. To the best of our knowledge, this is the first paper that systematically reports the advances on models and applications of multi-way analysis in NIRS for the food industry.ith column of matrix i, j and k mean the indexes belonging to each of three modes of In order to avoid confusion, we will use the conventional and standard notations from the multi-way analysis community. Consistent with the notation used in Kiers , we use product . All theN-way partial least squares (N-PLS) is a regression algorithm combining tri-linear decomposition and the classical PLS. In fact, it is an extension of the two-way PLS model to the multi-way case. The real N-PLS model was proposed by Bro in 1996,rameters . In orderameters later prThe difference with the old N-PLS model is the core array o et al. . The so-Compared with the two-way PLS model, the N-PLS model retains the multi-way information of the data, avoids the huge number of parameters caused by unfolding the multi-way array and the difficult model interpretation caused by the confounding of modes. It is capable of identifying multi-way data patterns and complex feature correlation. Apart from these advantages, N-PLS also has advantages in better modeling accuracy, robustness to noise, stabilized solution, increased predictability, etc. . Some prkth slab of three-way array kth row of matrix kth observation of the third mode, and fth column vector of loading matrixes PARAFAC was first proposed by Harshman in 1970.k-ranks of matrixes One of the most attractive advantages of PARAFAC is the uniqueness of the solution , which mnd Kiers . Like mand Kiers . Furthernd Kiers proposednd Kiers ,51.Even though PARAFAC has gained extensive acceptance nowadays, there are still some issues hampering the use of PARAFAC in practice. For example, multi-way analysis and data arrangement experiences are needed; such knowledge can be challenging for people who are accustomed to using only two-way chemometrics tools. Apart from this, algorithm problems are maybe another major concern. For instance, slow convergence occurs especially when facing seriously collinear data. The two-factor degeneracy problem, which means two components become almost identical but with opposite signs, is also a practical concern. In order to improve the convergence speed of the conventional PARAFAC-ALS algorithm, a wide range of remedies have been developed, such as line search , enhancek of the\u2014in this case\u2014third mode. Thus, the strict tri-linearity assumption is actually relaxed in one mode in the PARAFAC2 model. Compared with the assumption of PARAFAC which is requiring k, only the cross products of k in the third mode in PARAFAC2 [PARAFAC2 was also proposed by Harshman in the 1PARAFAC2 . This lePARAFAC2 . The conal paper . Hence, rvations . UnderstOwing to the advantage of the less strict tri-linearity requirement, which is often more realistic in the real world, PARAFAC2 is widely applied for solving a wide range of complex data analysis problems, e.g., the problem of varying batch trajectories in food process analyses (different batches have different numbers of NIRS spectra) or analyzing complex NIRS image data. As an important multi-way analysis model, PARAFAC2 has all the advantages of the general PARAFAC model. One of the challenges of the PARAFAC2 algorithm is how to impose the non-negativity constraints on all three modes. For the conventional PARAFAC2-ALS algorithm, it is not possible to impose a non-negativity constraint on the shifted mode, thus only two modes can be constrained. Recently, Cohen and Bro proposedThere are many alternative multi-way analysis models available for analyzing multi-way dataset. For instance, Tucker3 is a multi-way model of the Tucker family, also called the three-mode principal components analysis ,69,70. CPreprocessing plays an important role in chemometrics; however, it is more complicated when used for a multi-way array compared with two-way cases . Some sto et al. , single-o et al. , it is no et al. ; hence, o et al. , Inverseo et al. , Extendeo et al. and SNV o et al. are commo et al. are the o et al. ,81.Food industrial processes and food productions often involve multi-way data. A number of reports have shown the great potential of multi-way analysis tools for analyzing high dimensional and complex food process data ,83,84,85NIRS is widely used for the purpose of food quality evaluation and fraud detection ,94,95. MIdentification and classification are important application fields of NIRS and chemometrics , and areMulti-way analysis was also used for quantification and prediction purposes in the food industry . In partPenicillium digitatum in citrus fruits by analyzing the multi-way features array of the NIR hyperspectral image. It was reported that almost 91% of citrus fruit infected by Penicillium digitatum can be successfully detected by the multi-way model at early stages of the harvest, which will be of great importance for automating fruit sorting systems so that infected citrus fruits can be expelled before affecting the normal fruits. Yang et al. [Very little work has been done on a dedicated multi-way analysis for analyzing NIR image data even though multi-way methods have shown great potential for dealing with such types of structures. Folch-Fortuny et al. used an g et al. developeg et al. monitoreg et al. , huge amg et al. and consg et al. , cannot g et al. . By virtg et al. , the mulDuring the past decades, a number of software packages have been developed for performing multi-way analysis. One of the original software is the N-way toolbox for MatlThe application of multi-way analysis combined with NIR spectroscopy is still located at the initial stage for the food industry. So far, what we have seen about the synergy between rapid spectroscopic sensors and data analytic technologies, which has revolutionized the food industry, is only the beginning. Even though NIR spectroscopy can rapidly obtain thousands of data points in a short time, the potential of these big data sets has not been fully investigated. Deeper statistical and chemometric knowledge is desired by the food technologists, owing to the challenges of the high complexity of food processes and food products. As an advanced chemometric tool, multi-way analysis not only shows powerful advantages in food process analysis, quality evaluation, determination of chemical composition and structure, food image analysis, etc., but also makes the analysis process greener with the help of green and smart \u201cmathematical separation\u201d , fulfill"} +{"text": "Older adults, the largest segment of the US rural population, face significant disparities in health and healthcare compared to their non-rural peers, including more chronic health conditions, financial challenges, and social isolation. They have limited access to healthcare and social services for prevention, management and treatment of chronic conditions. Age-Friendly Care-PA, a partnership between Primary Health Network and Penn State College of Nursing, aims to reduce these disparities in care and services for rural older adults through co-designing their Geriatric Workforce Enhancement Program. Age-Friendly Health Systems, an initiative of the John A Hartford Foundation and the Institute for Healthcare Improvement, in partnership with the American Hospital Association and the Catholic Health Association of the United States, equips providers, older adults, and their care partners with the support necessary to address What Matters, Medication, Mentation, and Mobility. This symposium describes how the 4Ms are integrated into clinician training and competencies, older adult education, operations, care delivery, and quality improvement. Year two outcome data will be shared. Drs. Hupcey and Fick will provide an overview of the project and its reach. Dr. Berish will describe the process of engaging stakeholders in co-developing our 4M metrics and the data generated. Jenny Knecht, CRNP, will describe a pilot study to extend the reach and acceptability of telehealth to hard-to-reach older persons. Finally, Dr. Garrow will detail a new initiative focused on equity in care. Our discussant, Dr. Terry Fulmer will lead a discussion of this work as well as next steps and policy implications."} +{"text": "Due to the rapid increase in the use of electrical and electronic equipment (EEE) worldwide, e-waste has become a critical environmental issue for many governments around the world. Several studies have pointed out that failure to adopt appropriate recycling practices for e-waste may cause environmental disasters and health concerns to humans due to the presence of hazardous materials. This warrants the need for a review of the existing processes of e-waste management. In view of the growing e-waste generation in the Asia Pacific region and the importance of e-waste management, this study critically reviews previous research on e-waste generation and management practices of major e-waste producing nations in the Asia Pacific region, provides an overview of progress made and identifies areas for improvement. To fulfil the aims of this research, previous studies from 2005 to 2020 are collected from various databases. Accordingly, this study focuses on e-waste generation and environmental management of these countries. This study found that e-waste management practices of the selected countries need to be enhanced and recommends several best practices for effectively managing e-waste. The Asia Pacific region is highly populated and is considered one of the fastest developing regions in the world. In addition, many countries in this region underwent rapid industrialisation, driven by foreign direct investments due to aE-waste is one of the most urgent and pressing challenges of our time; however, it is routinely ignored. Across the world, the growing amount of e-waste threatens the environment and local communities, as incorrectly disposed e-waste results in life-endangering toxic chemicals released into the environment and the loss of precious metals ,4,5,6,7.This study identified three research gaps. Firstly, although, literature presents results of various studies on e-waste generation ,15,16,17The purpose of this study is to critically review the existing strategies and practices adopted by the major e-waste producing countries in the Asia Pacific region in managing and regulating e-waste to minimise the environmental and health impacts created as a result of inappropriate recycling and disposal practices.A key initiative and motivation of this study is to identify the problems/challenges in managing e-waste in the selected Asia Pacific countries and recommend appropriate management strategies and policy approaches to handle and regulate e-waste to significantly reduce environmental and health concerns. Accordingly, this study reviews previous research on e-waste generation and environmental management of Australia, China, India, Indonesia, and Malaysia, identifies problems and challenges that negatively impact e-waste management in these countries, provides an overview of progress made, and identifies areas for future research.The selected countries are among the largest producers of e-waste in the Asia Pacific region ,13,18,28This paper is organised into five sections. The first section presents current literature on e-waste, the research problem, research gaps and research aim, and justification for this study. The second section outlines the chosen methodology and the justification for considering a systematic literature review. The third section details the e-waste management practices in the selected countries. The fourth section provides the results of this study and analyzes the results. The final section presents the findings of this study, limitations associated with the current study, policy recommendations for effective e-waste management, and future research opportunities.In recent years, researchers have increasingly used quantitative and qualitative research (mixed methods) techniques to expand the scope and improve the analytic power of their studies ,30. QuanThis study adopts a qualitative research method to explore the issues relating to e-waste in the selected countries from existing research over the past years to guide future research in this area. To achieve the aim of this study, the five-phase approach of Wolfswinkel et al. for condThe first phase is to define the scope of the review. This includes the definition of specific criteria for the inclusion and exclusion of relevant sources and the criteria for identifying and retrieving those sources in the literature. In this study, four prominent databases are used to source literature, including ProQuest, Emerald, ScienceDirect, and Web of Science. The selection of these databases is due to their representativeness and coverage in the publication of top academic papers on e-waste in the selected countries. To ensure broad coverage of the studies in these databases, several keywords have been used for the search, which includes \u201celectronic waste\u201d, \u201ce-waste\u201d, \u201cwaste electrical and electronic equipment\u201d, \u201ce-waste management\u201d, \u201ce-waste recycling,\u201d \u201ce-waste disposal methods\u201d, \u201ce-waste problems and challenges\u201d and \u201cenvironmental management of e-waste\u201d. Several criteria are used to set the limitation, including restricting the document type to scholarly journals, peer-reviewed conference papers, book chapters, and other institutional reports from United Nations (UN) and World Health Organization (WHO); the language in English, and the publication date from 2005 to 2020. These document types have been selected as they represent state-of-the-art research outputs with high impact .The second phase is to run the search query within the selected databases for retrieving the search results. A total of 688 articles are returned using the above pre-defined search strings. This initial search enables us to gain a general understanding of the coverage of e-waste topics.The third phase involves selecting the final samples for detailed analysis. The search is limited to the title and the abstract to focus on the search results. Titles and abstracts of all initial articles are screened for checking the relevance to e-waste. This leads to the identification of 235 relevant articles. Duplicate articles are removed. A total of 210 articles is assessed for eligibility, and after excluding those articles that did not meet eligibility criteria, a total of 185 articles is identified for further review.The 185 articles have been read in full for coding and analysis. NVivo 12.0 is used for providing an overview of the general topics from all the abstracts of the included papers. An overview of the dispersion of the selected papers in terms of year of publication shows there is increased interest in e-waste from 2005 to 2020. E-waste is defined as an electrical appliance that no longer satisfies the user for its intended purpose . MeanwhiA further study was conducted in 2019 whereby the Asia Pacific region also generated the highest amount of e-waste in comparison to America, Europe, Africa, and Oceania regions. The Asia Pacific region generated around 25 Mt, followed by America at 13.1 Mt and Europe at 12.1 Mt. The study also showed that Africa generated 2.9 Mt and Oceania generated 0.7 Mt of e-waste . This waOver the years, the use of electronic devices for domestic and commercial purposes has grown rapidly . E-wasteThis study has selected five countries, including Australia, China, India, Indonesia, and Malaysia, from the Asia Pacific region because they are the major e-waste producers in the region. In line with the aim of this study, this section presents an in-depth analysis of waste generation, policies and management practices adopted by the selected countries in the Asia Pacific region. In addition, this section presents literature on e-waste generation and the opinions of scholars in this field. The following sub-sections explain e-waste management practices for the selected countries in the Asia Pacific region. Australia is placed among the top 10 consumers of electronic products in the world. As a result, e-waste has become one of the fastest-growing waste streams in Australia ,44,45. TThe introduction of the National Waste Policy in 2009 was designed to set the direction of Australia\u2019s e-waste management and resource recovery for 10 years from 2010 to 2020. The policy was established to achieve several goals, including compliance to international obligations such as the Basel and Stockholm Conventions, reducing the generation of e-waste, and ensuring e-waste treatment, disposal, recovery, and reuse is safe and environmentally sound ,47. The China is one of the leading producers of EEE, and currently, the country is experiencing incredible growth in e-waste generation from both domestic and international sources ,26,49. FInformal e-waste recycling in China is often carried out by individual recyclers and unauthorised dismantling companies. Informal recyclers purchase used items and often either dismantle or repair them for the second-hand market. This unregulated e-waste recycling method is currently flourishing in China. Informal recycling provides livelihoods for many Chinese citizens and is creating serious environmental and health concerns. Thus, e-waste generation and management in China has remained a major problem and are fuelled by China\u2019s inexpensive labour and manufacturing abilities. Informal recyclers do the majority of e-waste collection and recycling in most cities throughout China .The increasing average annual growth rate from 0.56% in 1991 to 1.62% in 2011 has contributed significantly to an alarming amount of e-waste generation in India. India is among the top 10 countries in the world in e-waste generation after the U.S. and China. It is estimated that three (3) million tons of e-waste were produced in 2018 and is expected to reach five (5) million tons by the end of 2020 ,52,53. AToday, e-waste in India is a significant waste stream both in terms of volume and toxicity . ApproxiThe Ministry of Environment and Forests (MoEF) is the national regulator responsible for formulating legislation related to e-waste management and environmental protection. MoEF approves the guidelines for the identification of the various sources of e-waste in India and endorses the procedures for handling e-waste in an appropriate and environmentally friendly manner . Those iDespite EPR being a major policy approach in both e-waste (Management and Handling) Rules 2011 and E-waste (Management and Handling) Rules 2016, they are not effectively implemented, and this can be attributed to certain peculiarities in India\u2019s e-waste management system ,61. For Due to substantial growth in the economy coupled with rapid technological developments, e-waste generation in Indonesia has increased considerably ,65. In 2Although the country has no presence of a specific regulation to manage its e-waste, the \u201cEnvironmental Protection and Management Act No. 32/2009\u201d and \u201cSolid Waste Management Act No. 18/1999\u201d are used in the regulation of e-waste produced in the country ,71. SincIn 2019, the International Monetary Fund (IMF), in its economic outlook, ranked Malaysia as the 3rd largest economy in Southeast Asia and the 37th largest economy in the world . With a Management of e-waste in Malaysia is still in its infancy and only began in 2005 . In MalaThis study considered literature reviews to identify key issues associated with e-waste management and to conduct an extensive evaluation of e-waste management practices in the selected countries. We believe this knowledge will help the countries to overcome their challenges and develop appropriate strategies for recycling and disposing of e-waste. This section provides an overview of earlier studies in the selected countries. In particular, results from the literature review on e-waste generation and management practices adopted by the respective nations are presented. Furthermore, this section presents the scope and the context of earlier studies on e-waste management. Prior studies ,84,85,86This study adopts a qualitative approach for studying e-waste management practices of the selected countries in the Asia Pacific region. As per Wolfswinkel et al. , this stE-waste management has become a contentious issue due to the presence of hazardous materials and the health hazards it may cause if not managed properly. In fact, for more than a decade, scholars have conducted studies on informal e-waste collection and disposal methods ,88. HoweAfter a critical review of the pertinent literature and a content analysis of the e-waste articles related to the selected countries, the dispersion of e-waste research in the selected countries according to the keywords/themes, e-waste categories examined, and the study location are illustrated in Given the background review and analysis in the previous sections, it is obvious that the problem and challenges of e-waste in the selected countries still persist. Our analysis shows that the e-waste management systems and infrastructure of the selected countries, particularly India, China, Malaysia, and Indonesia, are still in their infancy. Currently, e-waste scrap such as printed circuit boards, CRT monitors, and LCD screens have been, and are still being, recycled in China, India, Indonesia, and Malaysia, creating huge environmental and health issues. Informal e-waste collection, recycling, and its health implications on informal workers in these countries have become increasingly popular in the last 15 years ,92,93,94In China, several towns have remained as a dumping ground for e-waste. For example, Guiyu town is often referred to as \u201cthe e-waste capital of the world\u201d and employs more than 150,000 locals from four villages. These local informal workers dismantle and recapture valuable metals and parts that can be reused or sold from old computers. In Guiyu, it is not uncommon to see computer parts, cables, and huge tangles of wires scattered around the streets and riverbanks ,95,96,97In India, obsolete computers from households and businesses are sold by auction to door-to-door collectors who engage in informal methods of recycling. According to a report by the Confederation of Indian Industries (CII), approximately 146,000 tons of obsolete EEE are generated in India annually ,109. TheIn Indonesia, large amounts of e-waste are imported from developed countries. E-waste in the form of scrap materials or second-hand devices is sent to Indonesian islands from the adjacent ports in Singapore and Malaysia. Findings indicate that, in Indonesia, infrastructure and workable systems to quantify, recycle, monitor, and handle e-waste is lacking ,127. CurThe management of e-waste in Malaysia is still developing and only began in 2005 . ResultsIn Australia, several government policies have been developed. The key issues are identified in the e-waste management including: (a) the narrow scope of e-waste categories for recycling, (b) the lack of clarity on the roles of key stakeholders involved, (c) the recycling and material recovery targets, and (d) the lack of auditing and compliance. The results of the analysis show ,142,143 It can be seen that the majority of the selected countries in this present study are faced with an increasing amount of e-waste. Although the per capita e-waste generated in the emerging countries is much lesser than in the developing countries, the volume generated is greater due to the growing population and market size in emerging countries such as India, China, and Indonesia. These countries are ranked among the top e-waste generators in the world.The importance of selecting these countries such as Australia, India, China, Indonesia, and Malaysia in the Asia Pacific region in terms of environmental and market perspectives cannot be overemphasised. These selected countries have significant population, natural resources, and financial potentials ,150,151.Clearly, e-waste management processes in the majority of these countries examined still need improvement. Most of these countries studied have no well-established e-waste infrastructure for efficient collection, storage, transportation, recycling, and disposal of e-waste. In addition, the enforcement of codes of practice and regulations relating to hazardous e-waste management in these countries is minimal or non-existent.Exposure to e-waste is harmful to public health. E-waste has been found to negatively impact public health because communities are exposed to a complex mixture of chemicals from multiple sources and through multiple exposure routes . The resHence, e-waste has become one of the major challenges in these countries, and it is, therefore, crucial for these countries to investigate the development of a well-organised and inexpensive recycling scheme to extract valuable resources with inconsequential environmental impacts.This study has evaluated the e-waste generation and management practices of the selected countries in the Asia Pacific region. Based on the review of past studies and results of the analysis, it is obvious that the majority of the selected countries are yet to find a workable e-waste management strategy that will provide a sustainable solution to their e-waste concerns.Results of the analysis show that the volumes of e-waste generated are fast exceeding the available infrastructure and recycling facilities in the countries examined, thereby driving e-waste streams to flow into illegal and informal recovery. On top of that, the absence of an integrated framework that could support the monitoring and management of toxic and hazardous wastes has also created additional problems in managing e-waste in the selected countries and calls for a generic e-waste policy approach.In addition, the increasing demand for second-hand EEE, particularly in developing countries due to poverty and the continuing technological modernisation, has made these countries dumping grounds for e-waste from developed countries. For example, China\u2019s Guiyu town is well-known for the informal recycling of printed circuit boards. Specifically, \u201cmetal-contaminated sediments and elevated levels of dissolved metals have been reported in rivers around the town of Guiyu\u201d .Furthermore, sophisticated facilities and infrastructure required for formal recycling of e-waste using efficient technologies are minimal or non-existent in the selected countries. Formal recycling is widely accepted as the best way to manage e-waste, which reduces greenhouse gas emissions and helps lessen the climate crisis. Thus, recycling e-waste will reduce air and water pollution associated with the illegal dumping of e-waste. By recycling discarded, unwanted, or obsolete EEE for new products, nations can further reduce the enormous health risks and environmental pollution associated with improper disposal of e-waste.Therefore, to effectively manage e-waste in the selected countries, there is a need to develop generic structured policy approaches to tackle the e-waste problem in the selected countries and indeed across the world is required. These structured policies are projected to put in place formal systems and infrastructure for the recycling, management, and disposal of e-waste, taking into account country-specific issues.One of the shortcomings of this study is that the information and analysis of previous studies are seen to be reality. This study is also limited to countries in the Asia Pacific region and considers the time limitation by the year of the articles found. Although the accuracy of some of the analyses in the present study is inescapably subjective, this study is a starting point for further research into various aspects of e-waste generation and management practices of the selected countries.E-waste regulations tailored to each country\u2019s current situations should be enacted, recognising the lessons learned from more developed and experienced nations such as Japan, Switzerland, and South Korea;Extended producer responsibility (EPR) and 3Rs strategy should be implemented in EEE manufacturing regulations in all countries to support the production of simple, lightweight products, planned for reuse rather than obsolescence so that recycled materials can become resources for new products, thereby reducing the request for raw materials;Local government councils are key stakeholders in the management and recycling process and therefore incur major expenditures while handling e-waste. This, therefore, necessitates policymakers understanding of the determinants, drivers, and costs associated with e-waste collection and disposal;International integrated organisations should be established for checking specific e-waste material generation across the globe. This initiative will restrain the transboundary movement of e-waste across international borders.This study has exposed the current situation of e-waste generation and management practices of the selected countries. The following recommendations are suggested based on the findings of this study:Although different countries have endorsed and passed their respective e-waste regulations in other to manage e-waste, implementing appropriate and structured policy approaches will support all efforts directed towards effectively managing e-waste across the globe. Firstly, it is critical to have stepwise, and well-thought-out policy approaches for effectively formulating and implementing e-waste regulations and guidelines. Such approaches have been found to be effective in more advanced countries such as Switzerland, South Korea, and Japan, as noted above. In view of the multidimensional socio-economic nature of emerging economies, it is vital to consistently assess and evaluate existing policies to identify gaps and areas for improvement. This technique has also been found to be effective in Australia. Secondly, when implementing e-waste policies, interdisciplinary research approaches need to be considered. This will allow policymakers to better understand and address the various health and environmental problems associated with e-waste management. Finally, we believe that the policy approaches of respective countries geared towards dealing with the persistent and challenging e-waste issues require a local and specific approach where inherent socio-cultural, economic, political, and environmental concerns of that country are taken into consideration.Future research should use a quantitative approach or other research methods and expand the number of selected countries to understand e-waste generation and management practices of countries in the Asia Pacific region. This will provide additional viewpoints in the management, recycling, and environmental management of e-waste in the regions."} +{"text": "To assess the impact of retinopathy and systemic vascular comorbidities on the all-cause mortality in a representative U.S. sample.st, 2015), whichever came first. Risks of mortality were estimated using Cox proportional hazards models after adjusting for confounders and vascular comorbidities.A total of 5703 participants (\u226540 years old) from the 2005-2008 National Health and Nutrition Examination Survey. The Early Treatment Diabetic Retinopathy Study grading scale was used to evaluate the retinopathy status. Systemic vascular comorbidities included diabetes mellitus (DM), high blood pressure (HBP), chronic kidney disease (CKD) and cardiovascular disease (CVD). Time to death was calculated as the time from baseline to either the date of death or censoring (December 31After a median follow-up of 8.33 years (IQR: 7.50-9.67 years), there were 949 (11.8%) deaths from all causes. After adjusting for confounders, the presence of retinopathy predicted higher all-cause mortality (hazard ratio (HR), 1.41; 95% confidence interval (CI), 1.08-1.83). The all-cause mortality among participants with both retinopathy and systemic vascular comorbidities including DM , HBP , CKD and CVD was significantly higher than that among those without either condition. When stratified by diabetic or hypertension status, the co-occurrence of retinopathy and CKD or CVD further increased the all-cause mortality compared to those without either condition.The co-occurrence of retinopathy and systemic vascular conditions predicted a further increase in the risk of mortality. More extensive vascular risk factor assessment and management are needed to detect the burden of vascular pathologies and improve long-term survival in individuals with retinopathy. Retinopathy commonly refers to a spectrum of signs on the fundus that are common in the elderly, even in those without diabetes . A recenPrevious literature has consistently described significant associations between retinopathy and systemic vascular comorbidities, including diabetes mellitus (DM) , high blvia non-invasive retinal imaging and that there is a well-established association between retinopathy and systemic vascular comorbidities, clarifying the precise effects of the co-occurrence of retinopathy and systemic vascular comorbidities on the risk of mortality is of great significance.To date, only a few studies have investigated the joint effects of retinopathy and systemic vascular comorbidities : 7.50-9.67 years). There were 949 (11.8%) deaths from all causes. Participants with retinopathy had higher all-cause mortality than those without . Kaplan-Meier curves for all-cause mortality according to retinopathy and systemic vascular conditions are shown in The results stratified by diabetes and hypertension status are shown in In this large sample of middle-aged and older adults, participants with retinopathy had a higher all-cause mortality rate. Moreover, the co-occurrence of retinopathy and systemic vascular conditions further increased all-cause mortality.Our findings are in line with previous results that reported a significant association between retinopathy and mortality \u201326. ContOnly a limited number of studies have attempted to explore the joint effects of retinopathy and systemic vascular comorbidities [such as DM , CKD 1616, 29\u201331Mechanisms underlying the association between retinopathy and mortality remain unknown. It has been proposed that retinopathy may be an indicator of the burden of CVD risk factors, including dyslipidemia, obesity, hypertension, and smoking . Howevervia non-invasive means, our findings may have several practical implications. Firstly, our finding of a statistically significant association between retinopathy and systemic vascular abnormalities indicates that individuals with retinopathy may benefit from a comprehensive vascular assessment and should be closely monitored. Secondly, the current study and previous studies find that the co-occurrence of retinopathy and micro- or macrovascular disorders poses a further increase in all-cause mortality. Therefore, intensive vascular risk reduction may be warranted in the management of these patients.Given that retinopathy can be readily detected The strengths of this analysis include the use of a large-scale sample with national representativeness, the use of a standardized objective grading protocol to assess retinopathy, access to death records, long-term follow-up and inclusion of a comprehensive range of covariates. However, several potential limitations should be considered. Firstly, retinopathy status and potential confounders adjusted for in our analysis were assessed on a single occasion. During the follow-up period, the behaviors and/or retinopathy status of the patients may have changed, which may have a direct impact on the outcome. Secondly, discrepancies may exist between the self-reported data and clinically measured data. This may cause underreporting of milder cases and subsequently lead to a bias in the analysis. Finally, participants with missing data tended to be in poorer health status, which may have resulted in selection bias.In summary, our findings suggest that middle-aged and elderly people with retinopathy have increased all-cause mortality. Furthermore, the joint effects of retinopathy and major systemic vascular comorbidities increase the all-cause mortality further. Our results indicate that more extensive risk factor assessment and management of individuals with retinopathy may be beneficial to reduce their mortality rate, especially in patients who have both retinopathy and systemic vascular disorders.https://www.cdc.gov/nchs/nhanes/index.htm.Publicly available datasets were analyzed in this study. This data can be found here: Study concept and design: ZZ, MH, and XY. Acquisition, analysis, or interpretation: All authors. Drafting of the manuscript: ZZ and XS. Critical revision of the manuscript for important intellectual content: WW, JH, YC, MH, and XY. Statistical analysis: ZZ, XS, and WW. Obtained funding: MH. Administrative, technical, or material support: ZZ, XS, WW, MH, and XY. Study supervision: MH and XY. All authors contributed to the article and approved the submitted version.The present work was supported by Fundamental Research Funds of the State Key Laboratory of Ophthalmology (303060202400362), National Natural Science Foundation of China , Project of Special Research on Cardiovascular Diseases (2020XXG007), and Research Foundation of Medical Science and Technology of Guangdong Province (B2021237). MH receives support from the University of Melbourne at Research Accelerator Program and the CERA Foundation. The Centre for Eye Research Australia receives Operational Infrastructure Support from the Victorian State Government. The sponsor or funding organization had no role in the design or conduct of this research.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The growth of non-standard employment relations has created one of the major challenges in terms of workers' rights as well as collective representation in European societies. Among non-standard employment relations, so-called \u201csolo self-employed\u201d\u2014self-employed workers without employees\u2014are challenging the very foundations of our labor markets, that is to say the opposition between employers and employees, fostering the development of emerging \u201chybrid\u201d areas of work. The heterogeneity of the solo self-employed is difficult to capture from official statistics, which are still based on traditional classifications, and questions also the legal categories that qualify these workers. Moreover, the fact that solo self-employed workers do not form a homogenous group, and are diverse in terms of their activities, interests and needs, calls for changes in the way trade unions, employer organizations, and new freelancer associations develop collective actions, claims-making activities, and strategies of organizing. With the aim to achieve an in-depth understanding of the increasingly extensive and populated categories of the solo self-employed, this contribution aims at reconstructing the state of the art within different fields of study, such as employment relations, labor law, industrial relations and social movements, and at offering some possible future research directions. The growth of non-standard employment relations has created one of the major challenges in terms of workers' rights as well as collective representation in European societies Cordova, . Among nThe heterogeneity of the solo self-employed is difficult to capture from official statistics, which are still based on traditional classifications, and also questions the legal categories that that these workers qualify for 960489;The Research Ethics Committee of the University of Leeds on 13/09/2017, ethics reference LTLUBS-175;The Research Ethics Committee of the University of Milan on 08/11/2018, ethics reference No. 50/18;The Data Protection Officer of the University of Milan on 10/11/2018. The patients/participants provided their written informed consent to participate in this study.The ERC ethics officer approved all the new elements submitted for ethics on 28/05/2019.This paper is an entirely collaborative effort by the all authors. If, however, for academic reasons individual responsibility is to be assigned. AM: wrote introduction and our proposal for a future research agenda. RB: trends and heterogeneity of solo self-employment. PD: the self-employment in the frame of labor law. PM, MM-N, and PB: the collective representation of solo self-employed workers.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Background: Intertrochanteric femur fractures, which are common geriatric osteoporotic fractures, have imposed a huge economic and social burden. This study clarified the global status of research on intertrochanteric fractures between 2001 and 2020 and predicted future research trends in this field using bibliometric and visualized studies.Methods: Publications related to intertrochanteric fractures were retrieved from the Web of Science (WoS) database. All studies were published between 2001 and 2020. Bibliometric and co-occurrence analyses were conducted using VoS viewer software.Results: In total, 2,632 studies were retrieved. The number of global publications regarding intertrochanteric fractures increased annually. The United States was the largest contributor, ranking first in total publications, citations, and the H-index. Switzerland had the highest average citation frequency among the 10 countries with the highest number of publications. The journal that published the most articles regarding intertrochanteric fractures was the Injury International Journal of The Care of The Injured, with 290 articles published. This journal also ranked first in the citation frequency. MJ Parker, an author, published the most papers in the field, and the University of California research team at San Francisco contributed the most publications in this field. During the co-occurrence analysis, all keywords were divided into four clusters: internal fixation study, complication study, risk-factor study, and survival and prognosis analysis study. The internal fixation and survival and prognosis analysis studies were predicted as the next hot topics in the field of intertrochanteric fractures.Conclusions: Intertrochanteric fractures are gaining increasing research attention according to the current global trend, and the number of publications regarding intertrochanteric hip fractures will continue to increase. The United States currently publishes the most articles on intertrochanteric fractures. The number of studies related to internal fixation, survival, and prognosis analysis is increasing, suggesting that these topics may become the next research hotspots in the area of intertrochanteric fractures. Intertrochanteric femur fractures, the most common type of fragility fractures in the elderly, account for 55% of proximal femoral fractures . As lifevia internal fixation with a dynamic hip screw (DHS), percutaneous compression plate (PCCP), proximal femoral locked compression plate, less invasive stabilization system, or intramedullary fixation devices and Science Citation Index Expanded (SCIE) databases. The WoS and SCIE databases are the optimal databases for bibliometric data and include over 12,000 high-impact, quality scientific international journals. All the publications included in this study were articles or reviews published in English between 2001 and 2020 and were retrieved from the WoS database . The folThe relevant information of all identified publications, including title, keywords, abstract, year of publication, author, impact factor, H-index, nationality, affiliates, research direction, and funding agencies, was downloaded from the WoS database. Any publication for which the relevant information was not available was excluded from this study. Two authors independently conducted the search and gathered the relevant data. The final screening results were reviewed by a third author.All identified data were rearranged according to the publication year, country, number of publications, total citations, average citations per term, H-index, journal, author, institution, and research fund direction. The H-index is an author-level metric that measures the productivity and citation impact of publications by an individual author , 13. MicThe bibliometric visualization and analysis of the literature were conducted with the Java program VoS viewer . In this study, the VoS viewer was used to conduct the bibliometric analysis and visualization research co-citation, co-authorship, co-occurrence, and bibliographic coupling. The associations between authors, institutions, and countries were visualized using weighted total link strength (TLS) lines. The TLS represents the total strength of the links of an item with other items, and the higher the TLS, the more weight is used to draw the link on the visual analysis.The co-citation analysis method is a quantitative intelligence research method used to study the relationships between documents by analyzing the frequency of simultaneous citations of the two documents by other documents. The VoS viewer software is used to calculate the co-citation link strength between two items.The co-occurrence analysis determines if the relationship between the items is based on the number of publications in which they appear together . The purBibliometric coupling is a more advanced research method of literature coupling. If papers A and B cited the same references, they have a coupling relationship, indicating that their research contents are similar. The VoS viewer was used to analyze the contents of all selected literature.n = 281; 7.6%) . The pre; 13.0%) .n = 615; 23.5%) followed by China and England .n = 15,848), followed by England , Switzerland , Canada , and Germany . The publications from Switzerland had the highest average number of citations (n = 41.46), followed by Canada (n = 27.66), England (n = 25.6), and Germany (n = 23.54).The United States had the highest H-index (H-index = 60) followed by England (H-index = 36), Germany (H-index = 36), and Canada (H-index = 33). The United States also had the most total citations (n = 290) followed by the Journal of Orthopedic Trauma (n = 183), Archives of Orthopedic and Trauma Surgery (n = 102), and International Orthopedics (n = 94) followed by the Journal of Orthopedic Trauma , the Journal of Bone and Joint Surgery, American Volume , and Osteoporosis International . The Journal of Bone and Mineral Research had the highest impact factor , followed by the Journal of bone and Joint Surgery, American Volume (IF = 4.578) and Clinical Orthopedics and Related Research (IF = 4.329).Among the journals, the Injury International Journal of the Care of the Injured had the most publications regarding intertrochanteric fractures ((n = 94) . Injury n = 1,596; 60.6%), surgery , general internal medicine , and emergency medicine .n = 27), followed by Yong-Chan Ha (n = 24), Kyung-Hoi Koo (n = 20), Timo Jamsa (n = 19), and Young-Kyun Lee (n = 19) (Martyn J Parker had the most publications regarding intertrochanteric fractures ((n = 19) .n = 48), followed by the Hospital of Special Surgery in the United States (n = 30), and Shanghai Jiao Tong University in China (n = 27) . The10 in = 98; 3.7%), followed by the National Institutes of Health and the National Natural Science Foundation of China . Seven fA total of 150 authors had at least five publications. Yong-Chan Ha had the greatest TLS (TLS = 47), followed by Kyung-Hoi Koo (TLS = 45), Young-Kyun Lee (TLS = 43), Shi-Min Chang (TLS = 27), and Timo Jamsa (TLS = 27).A total of 227 institutions had at least five publications. The five institutions with the greatest TLS were the University of California at San Francisco (TLS = 76), University of Pittsburgh (TLS = 40), Oulu University Hospital (TLS = 36), University of Oulu (TLS = 36), and Chung-Ang University (TLS = 34).A total of 47 countries had at least five publications. The five countries with the greatest TLS were the United States (TLS = 206), England (TLS = 135), Germany (TLS = 133), Switzerland (TLS = 93), and Austria (TLS = 88).A total of 150 authors had at least five publications. The five authors with the greatest TLS were Martyn J Parker , Matthias Knobe , Jong-Keon Oh , and Kyung-Hoi Koo .A total of 83 journals had at least five publications. The five journals with the greatest TLS were Injury-International Journal of Care of the Injured , the Journal of Orthopedic Trauma , Archives of Orthopedic and Trauma Surgery , International Orthopedics , and the Journal of Bone and Joint Surgery, American Volume .A total of 227 institutions had at least five publications. The five institutions with the greatest TLS were the University of California at San Francisco , University of Oulu , Korea University , Oulu University Hospital , and University of Toronto .A total of 47 countries had at least five publications. The five countries with the greatest TLS were the United States , China , Germany , England , and Turkey .There were 320 journals that were cited at least 20 times. The five journals with the greatest TLS were the Journal of Bone and Joint Surgery, American Volume , Clinical Orthopedics and Related Research , Injury International Journal of Care of the Injured , the Journal of Orthopedic Trauma , and the Journal of Bone and Joint Surgery, British Volume .A total of 429 publications were cited at least 20 times. The four publications with the greatest TLS were Cool TLS = 4 TLS = 3 TLS = 2 TLS = 2.A total of 744 keywords were used at least five times . The keyThe keywords were then categorized based on when they appeared in the literature . Before Bibliometric and visualized studies show the current status and predict the future development trends in research fields. This study presents a comprehensive overview of the trends and development of scientific output for intertrochanteric femur fractures from 2001 to 2020. The number of articles related to intertrochanteric fractures has increased nearly 6-fold since 2001. It is estimated that the number of publications in this field will increase to ~400 by 2030, indicating that this field will continue to be a research hotspot. The increasing number of publications is related to the increasing economic and social burden of intertrochanteric fractures, which also indicates that the research in this field will be thorough and comprehensive.The H-index and the total number of citations are important indicators of the academic impact and quality of a publication . The UniInjury International Journal of the Care of the Injured, the Journal of Orthopedic Trauma, Archives of Orthopedic and Trauma Surgery, and International Orthopedics) were identified as having published the most studies on intertrochanteric fractures. The total number of publications by these four journals accounted for approximately one-fourth of all publications in this field, indicating that future findings in this field are likely to be reported in these journals. Among the 10 journals with the most publications, an increase in the total number of publications tended to reduce the journal's impact factor in the short term.Four journals in the past 20 years, and this was also the case in 2020 . The possible reasons are as follows: (1) Because of the limitation associated with some medical conditions and, many underdeveloped countries lack the registration systems suitable for patients with hip fractures. (2) Furthermore, most published publications are low-quality retrospective studies, and high-quality RCTs and other functional publications are relatively lacking. (3) In addition, the community is guilty of performing a lot of research on topics that do not really matter in the overarching scheme of the patient's quality of life.The research directions and hotspots in the field of intertrochanteric fractures were identified using the co-occurrence analysis. The keywords were categorized based on study type, and the most frequently used keywords were associated with internal fixation and survival and prognostic analysis studies. Therefore, these study types are the hotspots in current research. However, this result may also indicate that these topics require additional research.The overlay visualization map was color-coded by the VOS viewer based on the average year that the keywords appeared in publications , which iThe results of this study indicate that the number of publications regarding intertrochanteric fractures has increased in the past two decades and that this trend will continue in the future. Intertrochanteric fractures will continue to be a hot topic in the field of orthopedics. The bibliometric and visualized analyses provide investigators with data regarding the leading countries, authors, institutions, and publications in this field, which will help researchers quickly understand the most advanced research and discoveries in this field. In addition, the co-occurrence analysis and overlay visualization map present the research hotspots and future research directions, which may help funding agencies and public health policy makers make more decisions regarding intertrochanteric fractures.The bibliometric analysis included in this study is limited by the use of one database. The scopes of the major databases differ, which may lead to some publications being omitted from the analysis when only one database is used. Additionally, only English-language studies were included, which may lead to a language bias. The impact of more recent publications may have been underestimated due to the low number of citations. Therefore, researchers must be aware of the latest published literature and literature published in languages other than English.This study clarified the global research status for intertrochanteric fractures between 2001 and 2020 and predicted future research trends in this field. The United States contributed the most publications and citations, whereas the Injury International Journal of the Care of the Injured can be considered a landmark journal as it has published the most papers related to this field. Studies regarding the survival, prognosis, and mechanical failure of internal fixations will be the research hotspots in the future. In future research, we need to carry out more high-quality RCTs research. In addition, we call on countries such as Southeast Asia and Africa to establish standardized hip fracture registration systems as soon as possible, which will help further improve the global hip fracture prevention and treatment system.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.LW: project administration, validation data curation, writing\u2014original draft preparation, supervision, and funding acquisition. ZZ: conceptualization, methodology, data curation, formal analysis, and writing\u2014original draft. YQ: methodology, writing\u2014review, and editing. YWZ: data curation and writing\u2014original draft. YZ and FS: visualization and validation. JL: data curation, writing\u2014review, and editing. TZ: data curation and project administration. All authors contributed to the article and approved the submitted version.This study was supported by Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences (2021-JKCS-021).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Background: Data regarding delivery of evidence-based care to critically ill patients in Intensive Care Units (ICU) during the COVID-19 pandemic is crucial but lacking. This study aimed to evaluate the implementation rate of the ABCDEF bundle, which is a collection of six evidence-based ICU care initiatives which are strongly recommended to be incorporated into clinical practice, and ICU diaries for patients with and without COVID-19 infections in ICUs, and to analyze the impact of COVID-19 on implementation of each element of the bundle and independent associated factors.Methods: A world-wide 1-day point prevalence study investigated the delivery of the ABCDEF bundle and ICU diary to patients without or with COVID-19 infections on 27 January 2021 via an online questionnaire. Multivariable logistic regression analysis with adjustment for patient demographics evaluated the impact of COVID-19 and identified factors in ICU administrative structures and policies independently associated with delivery.Results: From 54 countries and 135 ICUs, 1,229 patients were eligible, and 607 (49%) had COVID-19 infections. Implementation rates were: entire bundle (without COVID-19: 0% and with COVID-19: 1%), Element A (regular pain assessment: 64 and 55%), Element B , Element C (regular sedation assessment: 45 and 61%), Element D (regular delirium assessment: 39 and 35%), Element E (exercise: 22 and 25%), Element F (family engagement/empowerment: 16 and 30%), and ICU diary (17 and 21%). The presence of COVID-19 was not associated with failure to implement individual elements. Independently associated factors for each element in common between the two groups included presence of a specific written protocol, application of a target/goal, and tele-ICU management. A lower income status country and a 3:1 nurse-patient ratio were significantly associated with non-implementation of elements A, C, and D, while a lower income status country was also associated with implementation of element F.Conclusions: Regardless of COVID-19 infection status, implementation rates for the ABCDEF bundle, for each element individually and an ICU diary were extremely low for patients without and with COVID-19 infections during the pandemic. Strategies to facilitate implementation of and adherence to the complete ABCDEF bundle should be optimized and addressed based on unit-specific barriers and facilitators. For patients in the intensive care unit (ICU), evidence-based treatment such as the ABCDEF bundle \u20134 and ICHowever, drastic changes in practice related to the COVID-19 pandemic, including unbalanced resources, overwhelmed facility capacity, and strict infectious regulations, occurred world-wide and prevented ICU staff from performing evidence-based approaches to patient care in ICUs . Our recTherefore, we conducted a 1-day point prevalence study, to investigate the implementation rate of evidence-based ICU care for both patients without and with COVID-19 infections and the impact of COVID-19 infections on implementation on a world-wide scale to capture the current clinical practice situation. We sought to identify ICU-related factors associated with implementation in the ICU.This was an international 1-day point prevalence study conducted on 27 January 2021, with approval by the ethics committee of the Saiseikai Utsunomiya Hospital (2020-69) and pre-registration in UMIN (ID: 000040405). The study design and construction followed the STROBE cross-sectional guidelines.https://form.jsea2005.org/isiic-II-study/), ethical considerations, and the URL for registration created by Google Forms (Google Inc.). According to the Ethical Guidelines for Medical and Health Research Involving Human Subjects in Japan is shown in https://datahelpdesk.worldbank.org/knowledgebase/articles/906519-world-bank-country-and-lending-groups) according to the country where the participating ICUs are located, which was obtained as the background data. Each representative received a different Facility Registration Number automatically soon after the completion of the questionnaire. On the survey date, 27 January 2021, the URL for the survey of evidence-based and supportive ICU care were sent to all registered representatives. All representatives were asked to input the institution-specific Facility Registration Number at the first question, and only those who had it could continue to complete the survey , and ICU length of stay as of the survey date, use of medical devices, continuous use of neuromuscular blockade, vasoactive, analgesia, and sedation agents, prone positioning and its duration, the presence of a target/goal of each ICU care modality given to ICU patients on the survey date, and the implementation of each element of the ABCDEF bundle, ICU diary provided on the survey data were collected. The operational definitions of each element of the ABCDEF bundle and ICU diary were proAll the data were stored online and managed or exported by the authorized person out of the authors .The primary outcome was the implementation rate of the entire ABCDEF bundle. Secondary outcomes were the implementation rates for each element of the ABCDEF bundle, including element A (regular pain assessment), element B [both spontaneous awakening trials (SAT) and spontaneous breathing trials (SBT)], element C (regular sedation assessment), element D (regular delirium assessment), element E (early mobility and exercise), and element F (family engagement and empowerment), and an ICU diary.The implementation of element E during mechanical ventilation, the implementation of element F performed online, and visitation policies for family members were also described. Independent factors associated with successful implementation of each element of the ABCDEF bundle were evaluated by multivariable logistic regression analysis.U-test for non-normally distributed continuous data and the chi-squared test and Fisher's exact test for categorical data appropriately. There was no missing data.Non-normally distributed continuous data were reported as medians with interquartile range (IQR). Categorical data were described as numbers or percentages. Comparisons of patient demographics, implementation of the ABCDEF bundle, and the ICU diary between the groups of patients with out and with COVID-19 infections were made with the Mann-Whitney In multivariable logistic regression analysis with adjustment for patient demographics, the association between the implementation of each element of the ABCDEF bundle and the presence of COVID-19 infection or ICU administrative structures was investigated. Patient demographics included length of ICU stay, age, gender, body mass index, use of mechanical ventilation, extracorporeal membrane oxygenation including veno-venous and veno-arterial, renal replacement therapy, and left ventricular unloading device, continuous use of neuromuscular blockade, vasoactive drugs, analgesia agents and sedation agents, and prone positioning. The following variables were changed to factors and used in the multivariable logistic regression analysis: number of hospital beds, nurse-to-patient ratio, frequency of multidisciplinary rounds, number of visiting hours for a family, type of hospital and ICU, primary responsibility to make decisions to implement the ABCDEF bundle, age, body mass index, income level. As a sub-analysis, associated independent factors among ICU administrative structures for each group (non-COVID-19 and COVID-19) were evaluated through the stepwise method with Akaike information criterion and with adjustments of the same variables of patients demographics described above. The stepwise method was used to focus on significant factors. In the sub-analysis, the variables that the number of patients allocated to the category is too few (\u22645 patients) to create a suitable model were excluded from multivariable logistic regression analysis.p-value was reported as two-sided and p <0.05 was considered statistically significant.The calculated sample size with 95% power and a two-sided alpha of 0.05 was 508 patients under the assumption of the implementation rate of the entire ABCDEF bundle for patients without and with COVID-19 infections based on previous surveys , 13. Allp = 0.004). Compared to before the pandemic, family visiting hours to patients both without and with COVID-19 infection were reduced , and more stringent restrictions imposed on families of patients with COVID-19 infections (p <0.001). A specific protocol for each element of the ABCDEF bundle was in place in <50% of ICUs except for a protocol for pain management (51%).Of 283 registered ICUs, 135 ICUs completed the survey (response rate 48%) , 2. RespThe details of the types of hospitals and ICUs participating, professionals dedicated to the ICU, and the personnel with primary responsibility for implementing the ABCDEF bundle are shown in There were significant differences in the demographics of the two groups for ICU length of stay, age, BMI, gender, use of mechanical ventilation (49 vs. 66%) and left-ventricular unloading device, continuous use of neuromuscular blockade, analgesia and sedation agents, prone positioning, and its duration . The twop = 0.53) (p = 0.59). Given elements A, C, D, E, and F which target all ICU patients, the implementation rate of all of these elements was low (1 vs. 3%. P = 0.07), even when one of the five was excluded .The implementation of the entire ABCDEF bundle, including elements A, B, C, D, E, and F which targets patients undergoing mechanical ventilation and continuous sedation, was rarely performed for patients both without and with COVID-19 infections . The ratElement A (64 vs. 55%), element B (17 vs. 10%), SAT (21 vs. 14%), and SBT (29vs. 16%) were implemented significantly less often for patients with COVID-19 infection, while element C (45 vs. 61%), element F (16 vs. 30%) and the online conduct of element F (4 vs. 21%), were performed significantly more frequently for patients with COVID-19 infections. There was no significant difference in the implementation of element D (39 vs. 35%), element E (22 vs. 25%), even while patients were undergoing mechanical ventilation (6 vs. 6%), and the ICU diary (17 vs. 21%). In-person visits were significantly less frequently allowed but online visits using electronic devices were more often used for the families of patients with COVID-19 infection.In multivariable regression analysis adjusted for baseline conditions, the presence of COVID-19 infection was not associated with non-implementation of individual elements of the bundle, but was significantly associated with implementation of elements D, E, and F .Among ICU administrative structural elements, specific factors associated with implementation were identified for each element of the bundle. Element A: presence of a written protocol and a target/goal, management as a tele-ICU, the presence of dedicated intensivists, and responsibility by a multidisciplinary team. Element B (SAT): presence of a written protocol, management as a tele-ICU, and responsibility by a multidisciplinary team and intensivists. Element B (SBT): management as a tele-ICU, responsibility by a multidisciplinary team, intensivists, and nurses, and being in an upper-middle-income country. Element C: presence of a written protocol and a target/goal, management as a tele-ICU, and presence of dedicated intensivists. Element D: management as a tele-ICU, performing multidisciplinary rounds daily and at least once a week or month, presence of dedicated respiratory therapists, and responsibility by nurses. Element E: presence of a target/goal and visiting hours (0 < x < 6 h). Element F: management as a tele-ICU, visiting hours (0 < x \u2264 24 h), presence of dedicated Intensivists, respiratory therapist, and nutritionist, and being in an upper-middle-income country.In the sub-analysis, a variety of different independent factors were identified for patients without and with COVID-19 infections . The preThis world-wide 1-day prevalence study demonstrates that implementation of the entire ABCDEF bundle, or its individual elements and an ICU diary for patients without and with COVID-19 infections is extremely low even though the implementation rate of specific individual elements of the ABCDEF bundle was different for the two groups. The presence of COVID-19 infection was not a factor preventing implementation. A variety of ICU-related factors were identified as independently associated facilitators or barriers for the implementation of the ABCDEF bundle, and these were different for each element, comparing patients without and with COVID-19 infections.Implementation of the ABCDEF bundle is much lower in this study compared with that reported by a survey before the pandemic , 21, butDifferences in the implementation of elements of the ABCDEF bundle might be caused by differences in underlying diseases and ICU length of stay between the two groups. Patients with COVID-19 infections, admitted to the ICU because of severe respiratory failure , need loAfter adjusting for the backgrounds of hospitals and ICUs and the baseline condition of patients, the presence of COVID-19 infection was not a barrier to the implementation of each element of the ABCDEF bundle. For elements D, E, and F, the presence of COVID-19 was significantly associated with their implementation. The warnings of high risk for and high incidence of delirium in the early stage of the pandemic may be a factor or the uThis study demonstrates the diversity of independent factors associated with the implementation of each element of the ABCDEF bundle in addition to variations comparing non-COVID-19 and COVID-19 settings. These results particularly show that a promising strategy to introduce or implement a specific element of the bundle in an ICU could vary and should be designed depending on the context and local situation in which it will be implemented. For many elements in the ABCDEF bundle, regardless of COVID-19 status, a specific protocol and presence of a target/goal for ICU care were consistently identified as facilitating independent factors. However, this study also showed the low frequency to equip the specific protocol in each ICU, or 50% or less, which could be considered as one of the major barriers to be managed regardless of the presence of COVID-19. As many studies successfully showed a pivotal role for implementation or introduction of ICU care, this simple, but not time- or resource-consuming approach could be a key stimulus and should be routine in the ICU to facilitate efficient implementation of evidence-based approaches to ICU care , 37, 38.This study has several acknowledged limitations. First, the limited number of patients and participating countries (Japan accounts for 40%) could lead to selection bias and limit generalizability to other ICUs and countries. These numbers might not be enough for the multivariable analysis with a number of covariates. Although the survey date captured a peak in the wave in Japan , the staThough having a COVID-19 infection was not associated with a failure to implement evidence-based ICU care, the implementation rates for the entire ABCDEF bundle, each of its elements and the ICU diary for patients without and with COVID-19 infections, were various, but extremely low on the whole regardless of the presence of COVID-19 infection. Since the impact of the COVID-19 pandemic on evidence-based ICU care varies depending on the conditions in each ICU, strategies to facilitate the implementation of each element of the ABCDEF bundle must be tailored to each institution.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Ethics Committee of the Saiseikai Utsunomiya Hospital. Written informed consent from the participants' legal guardian/next of kin was not required to participate in this study in accordance with the national legislation and the institutional requirements.KL, KN, HK, PN, EE, SK, KT, SI, and ON: study conception and design. KL, KN, HK, PN, EE, SK, and KT: statistical analysis or interpretation of data and drafting the manuscript. KN, HK, ME, PN, EE, SK, KT, MG, BL, CC, JB, SI, AL, and ON: critical review and revision of the manuscript for important intellectual insight. PN, EE, SK, KT, SI, AL, and ON: study supervision. KL, KN, HK, ME, PN, EE, SK, MG, BL, CC, JB, SI, and ON: recruitment the participating ICUs in overseas countries. KN confirmed that all authors meet authorship criteria according to ICMJE. All authors drafted the manuscript for important intellectual content, contributed to revision of the final version of the manuscript, approved the final version submitted, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.KL reports personal fees from MERA and receives a salary from TXP Medical completely outside the submitted work. KN reports personal fees from Abbott Laboratory, Nestle, TERUMO, GETINGE, Asahi Kasei Pharma, Ono Pharmaceutical, Japan Blood Products Organization, Nihon Pharmaceutical, Otsuka Pharmaceutical, Pfizer, Toray, and Baxter, and grants from Asahi Kasei Pharma outside the submitted work. HK receives a salary from the Japanese Society for Early Mobilization (non-profit society) as a chair (full time) outside the submitted work. EE reports grants from the VA/NIH; personal fees from Pfizer, Orion, and Lilly; personal fees from Masimo; and grants from Kohler outside the submitted work. SI reports personal fees from MERA, Abbott Laboratory, Teijin Pharma, Nestle, and Nihon Pharmaceutical. ON reports grants from Asahi Kasei Pharma, Ono Pharmaceutical, Baxter, Maruishi Pharmaceutical, Torii Pharmaceutical, Teijin Pharma, Shionogi Pharmaceutical, and Fuso Pharmaceutical outside the submitted work. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "BACKGROUND: We have been conducting a cohort study named \u201cthe Japan Collaborative Cohor Study (JACC Study) for Evaluation of Cancer Risk sponsored by the Ministry of Education, Science, Sports and Culture of Japan (Monbusho)\u201d since 1988. The aim of this paper is to describe the mortality of our JACC cohort in the follow-up period from 1988 through 1999, to compare it with the mortality, especially cancer deaths, of the Japanese population in the same period and to compare the causes of mortality by district among the cohort.METHODS: We conducted a follow-up study of 110,792 Japanese inhabitants aged 40-79 years in 1988-1990 for about 10 years to the end of 1999.RESULTS: Of 46,465 males, 37,750 (81.2%) were alive, 7,238 (15.6%) were dead and 1,477 (3.2%) had moved out of the study areas. The figures were 57,016 (88.6%), 4,940 (7.7%) and 2,371 (3.7%) among 64,327 females, respectively. The mean follow-up period was 9.9 years. The proportion of cancer deaths by site in our cohort members was almost same as the Japanese population aged 40-79 years old in 1995. Sex-specific standardized mortality ratios of total deaths, all cancer deaths, and most cancers in our cohort were less than 100 in both males and females for total cohort and the cohort by district.CONCLUSION: Our cohort members appeared to be almost the same or slightly healthier and less likely to die from total causes and cancers than the general population. The purpose of this study is to address the relationship between recent Japanese lifestyles and cancer. There were no other cohort studies in Japan at that time since Hirayama\u2019s large-scale cohort study2 on cancer which was initiated in 1965 and terminated in 1982, even though the Japanese lifestyle, especially dietary habits, have dramatically changed since the end of the Second World War in 1945.3 Epidemiologic studies using questionnaires on smoking, drinking and diet are important. However, those using biological markers can provide much more informative evidence for cancer pathogenesis. Therefore, another purpose of the JACC Study is to investigate the relationship between biological markers and cancer risk.A large-scale population based cohort study named \u201cthe Japan Collaborative Cohort Study (JACC Study) for Evaluation of Cancer Risk sponsored by the Ministry of Education, Science, Sports and Culture of Japan (Monbusho)\u201dThis paper will aim to describe the mortality of our JACC cohort in the follow-up period from 1988 through 1999, to compare it with the mortality, especially cancer deaths, of the Japanese population in the same period and to compare the causes of mortality by district among the cohort.The JACC study was conducted in 45 areas from 19 prefectures throughout Japan; 3 towns in Hokkaido district, 5 towns in Tohoku district, 5 towns in Kanto district, one city, 3 towns and 2 villages in Chubu district, 8 towns and 2 villages Kinki district, one city and one town in Chugoku district, and 4 cities, 9 towns and one village in Kyushu district. A basic cohort population of 127,477 healthy inhabitants in the above areas responded to the baseline questionnaire in 1988-1990. Basic cohort members, including 46,465 men and 64,327 women aged 40-79 years at entry, were followed up for about 10 years to the end of 1999.4 for converting the ICD 9th code to the 10th.The follow-up survey was conducted using population registries in local municipalities to determine the vital and residential status of the cohort in each area. All the cases that moved out of the study areas were treated as censored cases. Five subjects who were expunged from their residence record by authorities were also treated as censored cases and included in cases that moved out for computing the cohort numbers by follow up status as of end of 1999. All deaths that occurred in the cohort were ascertained by death certificates from local public health centers in the study areas under the authorities\u2019 permission from the Director-General of the Prime Minister\u2019s Office . The underlying causes of death were coded according to the International Classification of Diseases and Injuries (ICD) 10th version by verifying computer-stored data in the Ministry of Health, Labour and Welfare with the permission. Those already coded according to the ICD 9th version (from the time of the baseline survey through 1994) and stored in the computer data-base in the Ministry of Health and Welfare were re-coded in 1999 according to the ICD 10th version (after 1995), using a specifically developed computer program5 From a practical reason, the mortality rates in 1989 were used for the follow-up data in 1988-1990, and by the same manner, the rates in 1992, 1995, and 1998 were used for each three years of follow-up. Confidence intervals of SMR were calculated according to chi-square distribution when the observation number was 10 or larger,6 and according to Poisson distribution when less than 10.7Follow-up condition by sex and age group at entry as of the end of 1999 was computed. For those dead, causes of death, especially of cancer deaths, by sex and age group at entry as of the end of 1999 were also computed. Sex-specific standardized mortality ratios (SMRs) were calculated using sex- and age-specific person-years of following-up and sex- and age-specific mortality rates for all Japan in 1988-1999.Our entire study design, which comprised singular and collective use of epidemiologic data and biological materials (serum only), was approved in 2000 by the Ethical Board at Nagoya University School of Medicine, where the central secretariat of the JACC study is located.Total cancer deaths accounted for 38.7% and 35.0% in male and female total deaths as shown in Sex-specific SMRs of total deaths, all cancer deaths, and site-specific cancer deaths were shown in 1 The three commonest sites, cancers of the lung, stomach, and liver in males were of the same order and almost the same proportion among all cancer deaths as of the end of 1997.1 Among females, the three commonest sites, cancers of the stomach, large intestine, and lung were same in each site as of the end of 1997,1 but the proportion of cancer of the large intestine (12.3%) exceeded that of the lung (11.8%) if the cancer of the colon and rectum were combined.The follow-up condition of cancer deaths by site as of the end of 1999 was almost same as of the end of 1997.8 Because some of the cohort members were selected from participants in health check-ups or other kinds of screening, they might have had slightly healthier lifestyles that prevented them from dying with lifestyle related diseases such as cancers and cerebrovascular diseases. Cohort members of each district also appeared to be slightly healthier than the general population, as most SMRs of total deaths and site-specific cancer deaths by district were less than 100. It might be due to the small cohort size that some SMRs of site-specific cancer deaths by district were more than 100. Even though our cohort members were slightly healthier than the general Japanese population in the study period, internal comparisons between an exposed group and a group of unexposed to any factors within the cohort can also be justified as a cohort study.SMRs of total deaths, all cancer deaths, and most site-specific cancers were less than 100 in both males and females. This means that our cohort members appeared to be less likely to die from total causes and cancers in comparison with the Japanese population as observed other Japanese cohort.The present investigators involved, with the co-authorship of this paper, in the JACC Study and their affiliations are as follows: Dr. Akiko Tamakoshi (present chairman of the study group), Nagoya University Graduate School of Medicine; Dr. Mitsuru Mori, Sapporo Medical University School of Medicine; Dr. Yutaka Motohashi, Akita University School of Medicine; Dr. Ichiro Tsuji, Tohoku University Graduate School of Medicine; Dr. Yosikazu Nakamura, Jichi Medical School; Dr. Hiroyasu Iso, Institute of Community Medicine, University of Tsukuba; Dr. Haruo Mikami, Chiba Cancer Center; Dr. Yutaka Inaba, Juntendo University School of Medicine; Dr. Yoshiharu Hoshiyama, Showa University School of Medicine; Dr. Hiroshi Suzuki, Niigata University School of Medicine; Dr. Hiroyuki Shimizu, Gifu University School of Medicine; Dr. Hideaki Toyoshima, Nagoya University Graduate School of Medicine; Dr. Shinkan Tokudome, Nagoya City University Graduate School of Medical Science; Dr. Yoshinori Ito, Fujita Health University School of Health Sciences; Dr. Shuji Hashimoto, Fujita Health University School of Medicine; Dr. Shogo Kikuchi, Aichi Medical University School of Medicine; Dr. Akio Koizumi, Graduate School of Medicine and Faculty of Medicine, Kyoto University; Dr. Takashi Kawamura, Kyoto University Center for Student Health; Dr. Yoshiyuki Watanabe, Kyoto Prefectural University of Medicine Graduate School of Medical Science; Dr. Tsuneharu Miki, Kyoto Prefectural University of Medicine Graduate School of Medical Science; Dr. Chigusa Date, Faculty of Human Environmental Sciences, Mukogawa Women\u2019s University ; Dr. Kiyomi Sakata, Wakayama Medical University; Dr. Takayuki Nose, Tottori University Faculty of Medicine; Dr. Norihiko Hayakawa, Research Institute for Radiation Biology and Medicine, Hiroshima University; Dr. Takesumi Yoshimura, Institute of Industrial Ecological Sciences, University of Occupational and Environmental Health, Japan; Dr. Akira Shibata, Kurume University School of Medicine; Dr. Naoyuki Okamoto, Kanagawa Cancer Center; Dr. Hideo Shio, Moriyama Municipal Hospital; Dr. Yoshiyuki Ohno, Asahi Rosai Hospital; Dr. Tomoyuki Kitagawa, Cancer Institute of the Japanese Foundation for Cancer Research; Dr. Toshio Kuroki, Gifu University; and Dr. Kazuo Tajima, Aichi Cancer Center Research Institute."} +{"text": "Medical, health and wellness tourism and travel represent a dynamic and rapidly growing multi-disciplinary economic activity and field of knowledge. This research responds to earlier calls to integrate research on travel medicine and tourism. It critically reviews the literature published on these topics over a 50-year period (1970 to 2020) using CiteSpace software. Some 802 articles were gathered and analyzed from major databases including the Web of Science and Scopus. Markets (demand and behavior), destinations (development and promotion), and development environments (policies and impacts) emerged as the main three research themes in medical-health-wellness tourism. Medical-health-wellness tourism will integrate with other care sectors and become more embedded in policy-making related to sustainable development, especially with regards to quality of life initiatives. A future research agenda for medical-health-tourism is discussed. In 1841, Thomas Cook organized a tour of 570 people to travel from Leicester to Loughborough\u2019s hot springs . Thus, tToday, people continue to travel in the pursuit of relaxation, for health reasons, as well as fitness and well-being . As a reThe concept of medical-health-wellness tourism has emerged relatively recently as a scholarly field of enquiry in tourism ,5,6. AltA consensus is yet to be established on the definitions and contents of medical-health-wellness tourism, and how they interact, including their potential overlaps. Medical travel and tourism, health tourism, wellness tourism, and other similar terms tend to be investigated separately in tourism research ,18,19,20This study firstly reviews existing scholarly research through a meta-analysis of medical-health-wellness publications in the context of tourism . Then, tPrevious reviews of the literature and meta-analyses have contributed to clarifying the overall understanding of medical-health-wellness tourism. Existing literature reviews tend to be very broad, spanning health-oriented tourism, medical tourism, sport and fitness tourism, adventure tourism, well-being (Yang sheng in Chinese) tourism, cosmetic surgery tourism, spa tourism, and more.Medical tourism is an expanding global phenomenon ,23,24. DAnother set of authors defined health tourism as a branch of tourism in general in which people aim to receive specific treatments or seek an enhancement to their mental, physical, or spiritual well-being . This syWellness tourism is a key area of relevant research as well . One resAll in all, although earlier literature reviews provide invaluable insights into medical-health-wellness tourism, there is a lack of studies that approach this concept in a holistic way. This research seeks to redress this balance by delivering a holistic review of the literature with the following objectives in mind: (1) investigating international journal articles across the typologies of tourism outlined above; (2) identifying influential scholars that have significantly contributed to this field; and (3) summarizing key trends in markets, industry development and promotion, as well as policy-making and impacts. In order to achieve this, a systematic review was conducted to analyze research articles in medical-health-wellness tourism published over a 50-year period from 1970 to 2020.A two-step approach was adopted for the development of a database of publications for analysis with CiteSpace. The first step involved a search for relevant, high-quality refereed articles in medical-health-wellness tourism. Several academic journal databases, within tourism and hospitality but also including other disciplines too, were searched for relevant articles in medical-health-wellness tourism using a set of selected keywords. The ISI Web of Science and Scopus were chosen for this purpose as a result of their international recognition and comprehensiveness. Articles included in the list of references of selected articles were also considered valid as part of this search, in line with methodological suggestions for systematic literature searches . Cited aThe second step involved using appropriate, valid and representative search keywords. A total of 986 articles were gathered using the following keywords: medical tourism, health tourism, wellness tourism, and spa tourism. After careful sorting of these publications, using their abstracts and keywords, the number of articles in the database was narrowed down to 802. Of these, 615 were obtained using the keywords medical tourism or wellness tourism, 157 were located by searching for health tourism, and 30 were discovered using spa tourism as the search term. Using the above keywords and restricting the search to 50 years (1970\u20132020), the first article was found to be published in 1974. As a result, the ensuing analysis of the literature comprises the period from 1974 to 2020. The research tool used for this study was CiteSpace, which is a bibliometric analysis software developed by Professor Chaomei Chen of Drexel University based on the Java framework . This soThe data were classified and analyzed to achieve three specific goals. The first and primary goal of this review work was to analyze the content of the chosen articles, including year of publication, authors, journal impact factors, and the institutional affiliations of scholars in this field. The data were then sorted into categories. The order of authorship was not recorded. For multiple-authored articles, each author was given the same level of credit as sole authors. Second, one of the aims of this research was to discover associations in authorships, regions, and affiliations using statistical analysis. Third, the 802 articles were classified into dominant thematic categories applying the approach proposed by Miles and Huberman . Three fFinally, in order to refine the set of topic sub-categories, abstracts, first paragraphs, and conclusions were read to make the most appropriate assignments. This approach contributed to the more advanced stages of development of the classification of sub-categories and, consequently, the verification of findings.This section presents the results of the data analysis carried out in this study and provides further insights on the methodology adopted.The 802 articles selected were all published in English and in international peer-reviewed academic journals. Initially, the first stage of this literature search involved identifying academic journals publishing research articles on medical-health-wellness tourism. It was found that 38 articles had been published on this topic in Tourism Management, and 24 articles in Social Science & Medicine. Non-tourism journals in fields such as business, economics, and health, also contributed a significant number of publications in this field, as shown in The second aim was to identify the most prolific scholars in medical-health-wellness tourism research. This was achieved using co-occurrence network analysis of the authors of relevant research articles . Each noAmong the 2381 authors identified, 1820 (76.4%) contributed to only one article, whereas the remaining 561 (23.6%) authored two or more articles. The three most prolific authors were Jeremy Snyder, Valorie Crooks, and Rory Johnston.n =197). The second and third largest groups corresponded to Canada (n = 88) and the UK (n = 84), respectively, followed by Australia (n = 70) and South Korea (n = 65). As shown in Another objective was to illustrate the relationships and networks of authors publishing research on medical-health-wellness tourism. An analysis of countries this research originated from was carried out using the CiteSpace software. n = 47) were affiliated to Simon Fraser University in Canada. This university was followed by Sejong University in South Korea (n = 13), and the London School of Hygiene & Tropical Medicine (n = 13) in the UK. The top universities in terms of author frequency were based in Canada, USA, Australia, UK, South Korea, and Hong Kong.As shown in The fourth research objective was to elicit the prevailing research themes using the 802 articles gathered. First, an analysis of keyword frequency was performed to identify the main research interests. High frequency keywords reflect the research \u2018hotspots\u2019 in the field. Using CiteSpace\u2019s keyword visualization analysis function, the keyword co-occurrence knowledge map of medical-health-wellness tourism research was drawn to grasp the research \u2018hotspots\u2019 .Then, content analysis performed on the articles gathered for this study identified three main themes, namely: markets (tourist demand and behavior), destinations (development and promotion), and development environments (policies and impacts). An uneven distribution of research themes is highlighted in Previous studies have shown that the growth of medical-health-wellness tourism in developing countries is largely linked to lower costs, shorter patient waiting lists, and better quality of care . SimilarAs the demand for these forms of tourism has risen over time, processes and factors influencing decision-making have attracted growing levels of scholarly enquiry. For example, a political responsibility model was used to develop a decision-making process for individual medical tourists . A sequeMultiple factors may simultaneously influence decisions related to the destination for care, including culture , social Compared to other tourists, the mental activity and behavior of medical-health-wellness travelers are quite different. Medical tourists are less likely to question their need for surgery and tend to be much readier to accept it . The emoIn response to the demands of medical-health-wellness tourism, destination development and promotion are attracting growing levels of scholarly interest. Scholars from different countries have discussed the market status of Turkey ,59, the The advantages and disadvantages of Turkey were examined and indicated needs for improvements . In anotAs regards medical-health-wellness tourism destination development, scholars have explored research from various perspectives. Conceptual frameworks have been developed to include tourism destinations and services in the context of medical and health tourism ,67. AdviOnce a medical-health-wellness tourism destination is developed successfully, marketing and promotion are essential to attract tourists. As part of this process, informing potential patients about procedural options, treatment facilities, tourism opportunities, and travel arrangements are the keys to success . Most toNumerous businesses promote medical-health-wellness travel, including medical travel companies, health insurance companies, travel agencies, medical clinics, and hospitals . Among tScholarly research has also considered the factors that need to be taken into consideration in medical-health-wellness tourism promotion. This research has suggested that destinations should identify the specifics in their health tourism resources, attractions, and products, seek collaboration with others, and build a common regional brand . RegionaThe rise of medical-health-wellness tourism emphasizes the privatization of healthcare, an increasing dependence on technology, and the accelerating globalization of healthcare and tourism . There aWhile medical-health-wellness tourism is a potential source of revenue, it also brings a certain level of risk to destinations and tourists . The sprMedical-health-wellness tourism has emerged as a global healthcare phenomenon. Policy guidance is vital for the development of this sector in the future . There aThis study is based on a literature review of 802 articles on medical-health-wellness tourism from 1970 to 2020. Jeremy Snyder was found to be the most prolific author in this field with 45 articles. It has been found that the literature on this topic can be summarized into three themes: markets (tourist demand and behavior), destinations (development and promotion), and development environments (policies and impacts). The scholarly research in this growing field has undergone a shift in emphasis from tourist demand and behavior to the promotion and development of destinations, and, more recently, to policies and impacts. To attract more tourists, destinations should explore their potential for medical-health-wellness tourism. Accessibility, procedural options, treatment facilities, travel arrangements, safety guarantees, and government policies remain influential factors. In the development and promotion of this form of tourism, childhood vaccinations, oral health, legal frameworks, evaluation systems, entrance systems, and macro-policy continue to be areas of concern and where further research is required. Above all, meeting or exceeding tourist expectations and requirements is the most important consideration to promote medical-health-wellness tourism. Similarly, appropriate policy guidelines and frameworks are necessary to support this form of tourism. Importantly, medical-health-wellness tourism may result in negative impacts on the healthcare service provision for local residents in poorer countries, with tourists from richer countries benefiting to the detriment of local communities. However, if managed successfully, this form of tourism can also be a force for good in terms of fostering the economic development of countries delivering these services.The results indicated that the research literature is spread across a range of different disciplines and there is not one single venue for publishing in this field. A better integration of the research and improved understanding of the overlaps among medical, health, and wellness tourism is required.Medical-health-wellness tourism will, over time, integrate fully with other healthcare and wellness services. Similarly, medical challenges such as disease prevention and traditional medicine remain essential directions for the future of health tourism. This form of tourism will also integrate further with industries such as wellness culinary tourism, mindfulness tourism, active tourism (including adventure tourism), and even cosmetic surgery tourism, leading to a vast array of potential research avenues linked to health tourism destinations. These futures will greatly promote the physical and mental health of wellness tourists. This is another emerging direction for future medical-health-wellness tourism research.Medical-health-wellness tourism will become more significant forms of tourism and impact the development of different nations and areas. For example, this tourism will integrate with Chinese traditional culture. Traditional treatments and remedies will become more of an advantage and should be a topic for future medical-health-wellness tourism research, as well as in other countries with unique health cultures, treatments, and procedures. Thailand, Malaysia, and other Southeast Asian countries are favored by tourists from developed countries due to lower costs. In the future, these areas need to focus more on tourism product design, health tourism marketing, community participation, and cross-cultural communication. Developed countries such as the USA, Japan, and South Korea, will use advanced technology and medical equipment to take the path to high-end, high value-added tourism development. This will lead to some new research opportunities.Compared with other types of tourists, the needs of medical-health-wellness tourists will receive more attention. Based on previous research, the psychology and perceived value of these tourists are the focus of considerable research. In the future, more emphasis will be paid to people and especially to their psychological and physiological needs. Research on demand will become a more popular topic of this tourism research. Second, the current research on medical-health-wellness tourists is concentrated on the study of tourists in the USA and Canada. Future research should be more dispersed and diversified. Tourists from emerging countries such as Eastern Europe, Asia, the Middle East, and Africa will receive more attention.This study, inevitably, has a number of limitations, including the relatively modest amount of articles collected. Only articles written in English were considered. The sample number is rather small to represent the general research trends in medical-health-wellness tourism from 1970 to 2020. Therefore, it is desirable to increase the number of publications and expand the time and language coverage of the research articles to gain more insights.Although the research scope of medical-health-wellness tourism is vast, it lacks in-depth exploration. Current research is fragmented, lacks continuity and comprehensiveness, and therefore cannot be considered systematic. Also, the legal aspects of the development of this tourism, environmental capacity of medical-health tourism, wellness tourism management, and mechanisms of profit distribution for medical-health-wellness tourism are less frequently mentioned in research articles. Innovation in this field and international cooperation, and talent cultivation are also not sufficiently addressed. The methods used in medical-health-wellness tourism research are often simple. Scholars still use traditional descriptive statistics and related analysis methods. The theoretical foundation of medical-health-wellness tourism is still relatively weak. We are in the primary stage of this tourism research and in the development of related tourism products. People all over the world are eager for healthy lives. Medical-health-wellness tourism is likely to play a more important future role in travel medicine and tourism research. Beyond what has been done already, follow-up research should be focused on interdisciplinarity and based on the integration of industries. More theoretical research is necessary to support the future growth of medical-health-wellness tourism."} +{"text": "Handover Management (HM) is pivotal for providing service continuity, enormous reliability and extreme-low latency, and meeting sky-high data rates, in wireless communications. Current HM approaches based on a single criterion may lead to unnecessary and frequent handovers due to a partial network view that is constrained to information about link quality. In turn, HM approaches based on multicriteria may present a failure of handovers and wrong network selection, decreasing the throughput and increasing the packet loss in the network. This paper proposes SIM-Know, an approach for improving HM. SIM-Know improves HM by including a Semantic Information Model (SIM) that enables context-aware and multicriteria handover decisions. SIM-Know also introduces a SIM-based distributed Knowledge Base Profile (KBP) that provides local and global intelligence to make contextual and proactive handover decisions. We evaluated SIM-Know in an emulated wireless network. When the end-user device moves at low and moderate speeds, the results show that our approach outperforms the Signal Strong First and behaves similarly to the Analytic Hierarchy Process combined with the Technique for Order Preferences by Similarity to the Ideal Solution regarding the number of handovers and the number of throughput drops. SSF outperforms SIM-Know and AHP-TOPSIS regarding the handover latency metric because SSF runs a straightforward process for making handover decisions. At high speeds, SIM-Know outperforms SSF and AHP-TOPSIS regarding the number of handovers and the number of throughput drops and, further, improves the throughput, delay, jitter, and packet loss in the network. Considering the obtained results, we conclude that SIM-Know is a practical and attractive solution for cognitive HM. HM is responsible for making network (dis)connection decisions in a timely manner ,2. In thIn the networking literature, we find two types of approaches that address HM, namely, single criterion-based and multicriteria-based. Approaches based on a single criterion, such as the Signal Strong First (SSF), usually consider only the link quality in the end-user device for carrying out handovers. SSF compares the Received Signal Strength Indication (RSSI) of available networks and selects the network with the highest signal . Single This paper presents SIM-Know, an approach for improving HM. The contributions of SIM-Know are two-fold. SIM-Know proposes a Semantic Information Model (SIM) that allows us to make context-aware handover decisions by considering and relating criteria from several context information domains: Network, Application, User, UserDevice, and Handover. SIM-Know also introduces a SIM-based distributed Knowledge Base Profile (KBP) that offers local and global intelligence for making contextual and proactive decisions during the handover process. We evaluated SIM-Know in an emulated wireless network. When the end-user device moves at low and moderate speeds, the results show that our approach outperforms SSF (a single criterion approach) and behaves similarly to the Analytic Hierarchy Process combined with the Technique for Order Preferences by Similarity to the Ideal Solution , regarding the number of handovers and the number of throughput drops. SSF outperforms SIM-Know and AHP-TOPSIS regarding the handover latency metric because SSF runs a straightforward process for making handover decisions. At high speeds, SIM-Know outperforms SSF and AHP-TOPSIS regarding the number of handovers and the number of throughput drops and, further, improves the throughput, delay, jitter and packet loss in the network. From the obtained results, we conclude that SIM-Know is an attractive and feasible solution for cognitive HM.The rest of this paper is organized as follows: This section presents research on HM approaches based on both a single criterion and on multiple criteria. Next, we point out the main shortcomings of the related work. The work in employedHM allows an end-user device to keep an active connection when moving from one network\u2019s coverage area (BS or AP coverage) to another . HM compSIM-Know introduces SIM and KBP for improving HM. SIM allows SIM-Know to make context-aware handover decisions. In turn, the distributed KBP provides local and global intelligence to make rule-based cognitive decisions about network connection and disconnection. SIM and KBP envision diminishing the number of handovers and the number of throughput drops and, as a consequence, have a positive impact on several network performance metrics .Network, Application, User, UserDevice, and Handover) modeled by SIM. We use the CIM and the OWL to carry out SIM and user device resources, represented by the Resource class, which affects the QoS required by a user application . The Policy class represents the policies to apply to the Network class and governs the HM process. An example of a policy is to rank the candidate networks considering some criteria such as users\u2019 speeds and their movement patterns.Network superclass models the characteristics and status of a network by using the Topology, NetworkTraffic, and AccessPoint classes. The Topology class represents the network\u2019s organization, including nodes and links. The NetworkTraffic class represents data and control traffic passing by the network. The AccessPoint class models a networking device using wireless technology; this class considers the area covered by the Cell class, which includes the Range class, which contains the LargeRange and SmallRange classes. The isCoveredByCell, hasResource and belongToTopology properties represent the AccessPoint class\u2019s relationship with the Cell, Resource and Topology classes, respectively. The Resource class models the ability to manage the resource consumption of APs located at (Location class) a particular network point.The User superclass models the profile and behavior of the end-users by the UserPreferences, UserHistory, MobilityPattern and UserSpeed classes. The UserSpeed class includes the SlowMobility, ModerateMobility and HighMobility classes in order to model how fast an end-user moves. The UserPreferences class profiles the users with information related to, for instance, network preference by cost and service quality expectation. The UserHistory class models the historical (dis)connection of end-users. The MobilityPattern contains information about the end-users\u2019 mobility pattern, which is predictable from their trajectory and velocity. The Application superclass represents end-user applications with the ServiceProfile and QoS. The ServiceProfile class models the application\u2019s types . The QoS class allows the representation of a set of QoS requirements for each type of application.The UserDevice superclass models the end-user devices and their components by the DeviceProfile, DeviceStatus and Resource classes. The DeviceProfile class models the device\u2019s characteristics. The DeviceStatus class represents the device\u2019s current status . The Resource class models the ability to manage the resource consumption of end-user devices located at (Location class) a particular network point. The UserDevice superclass relates to the Application superclass via the runsApplication property, which allows knowledge of the applications that are running in each end-user device. The isUsedByUser property defines a relationship between UserDevice and User.The Semantic layer uses SIM (the entire model or a part) to obtain information from the data included in the Context layer. The Reasoning layer obtains knowledge from the information represented by SIM. The Adaption process acts on the layers to maintain updated data, information and knowledge. The Collaboration process enables the sharing of the obtained knowledge between KBP instances. Next, we detail the KBP\u2019s layers and processes.KBP is a distributed knowledge base that intends to provide local and global intelligence that supports making rule-based cognitive decisions about network connection and disconnection processes. Context layer includes contextual data about the user, network, device, application, and handover. Contextual data are essential for carrying out HM in environments with multiple wireless networks [Static Context sublayer involves data that do not change or rarely do; it plays a vital role in assisting with neighbor network discovery [Dynamic Context sublayer serves an updated network view, including dynamic data such as the application requirements of a device needing handover and capacity available in a target AP, which enables the upper layers (Semantic and Reasoning layers) to realize knowledge-based handovers.The networks . As in [networks ,33, thisiscovery . ExampleSemantic layer offers a SIM instance that is nourished by the bottom layer\u2019s contextual data. Thus, the Semantic layer structures the information to achieve intelligent, timely and context-aware HM (considering criteria from the Static and Dynamic contexts). For instance, the DeviceProfile, UserPreferences, UserSpeed and MobilityPattern SIM classes can be used to build up a map of candidate networks; overall, SIM classes provide a structure to contextual data. It is worth noting that we consider three KBP flavors depending on how they instantiate SIM. Network and Handover. Application, User, UserDevice and Handover. The Reasoning layer triggers the Initiation phase, selects the target network and realizes the handover itself by inferring knowledge from SIM. We use Description Logic (DL) [Reasoning layer and, so, HM; the reasoning rules generate local and global intelligence to make autonomous handover connection decisions. Each rule has a set of conditions and settings. To illustrate how the Reasoning layer operates, next, we present some of the rules that are modeled to realize a policy intended to select candidate networks proactively, considering the coverage of APs and the mobility pattern of end-user devices. For example, the Rule The gic (DL) to expreListing 1. Rule for APInRange.Listing 2 shows that the Rule rding to , thu canListing 2. Rule for UserSpeed.Listing 3 shows that Rule rding to , thr canListing 3. Rule for APRange.Listing 4. Rule for SoJournTime.Listing 5 shows that Rule Listing 5. Rule for CandidateAP.Listing 6. Rule for AssociateAP.Adaptation process allows SIM-Know to dynamically adapt to the changing environments and to enhance HM by modifying the content of the layers of the KBP instances. The content is modified in a bottom-up way, starting with the contextual data, followed by the SIM instances, and ending with the acquired knowledge when environmental changes happen, such as new networks appearing, dynamic traffic conditions and variations in QoS requirements. Furthermore, this process allows the addition and updating of the Reasoning layer seeking to meet QoS and to preserve network performance.The Collaboration process allows KBP . Each Subject is related to various pairs, Predicate\u2013Object. The Predicate identifies a property of the Subject while the Object provides the value of such a property . The Object can also be a ClassId, to represent relationships among Subjects.Notation , which eStatic context.Step 0: Each KBP gathers the Dynamic context, generate local knowledge, and exchange Measurement Reports by way of the Collaboration process. Source Step 1: Step 2: Source Step 3: Source Step 4: Target Step 5: Target Step 6: Source Step 7: Source Step 8: Step 9: Target Step 10: Step 11: Adaptation process.Step 12: Target The steps are as follows:This section presents the evaluation of SIM-Know in a WLAN, aiming to show its behavior regarding the number of handovers and the number of throughput drops, and its impact on various typical network performance metrics. We implemented the SIM-Know prototype for WLAN, including emulator . The quaIn the emulation experiments, scripts for generating traffic were developed by using the iPerf3 and D-ITrding to , AHP-TOPrding to . SecondlNext, we present how SIM-Know, SSF and AHP-TOPSIS impact various network performance metrics when the end-user device moves at PSIS see a. The paPSIS see b. The thPSIS see c. FigurePSIS see a. The jiPSIS see b. The thPSIS see c.Network, Application, User, UserDevice, and Handover). The KBP\u2019s Reasoning layer allows the achievement of cognitive HM. The distributed nature of KBP and its continuous updating allow the Handover Initiation phase to be proactive and operate with the local knowledge, built by We argue that the improvement in throughput, delay, jitter, and packet loss offered by SIM-Know is due to its context-aware, cognition and proactivity capabilities, which decreased the number of handovers and the number of throughput drops. In particular, SIM provides the context-aware capability to perform handover decisions through the comprehensive and semantic network view given by the information domains to afford the proactivity capability in HM. The evaluation results showed that, thanks to the aforementioned SIM-Know capabilities, our approach overcomes SSF regarding the number of handovers and the number of throughput drops when the end-user device moves at any speed and, further, equals AHP-TOPSIS when it travels at low and moderate speeds. SSF outperforms SIM-Know and AHP-TOPSIS regarding the handover latency metric because SSF runs a straightforward process for making handover decisions. SIM-Know overcomes AHP-TOPSIS regarding all evaluated metrics when the end-user device moves at a high speed, positively impacting the wireless network\u2019s performance in terms of the delay, throughput, packet loss and jitter metrics. Considering these results, we concluded that SIM-Know is an attractive and feasible solution for cognitive HM.For future work, we intend to enrich our approach with SDN and NFV capabilities to deal with the scalability issue imposed on HM by the Industrial IoT and the massive IoT 5G use case. We also plan to create an efficient model for communicating KBPs and evaluating SIM-Know when it makes handover decisions in scenarios with many end-user devices, high network traffic, and high load in APs."} +{"text": "Introduction: Ultimate Frisbee (UF) is a non-contact, challenging, and self-promoted team sport. Some factors such as the game environment and rules seem to influence athletes' behavior. Goals: Provide a robust systematic review (SR) of the psychological domains associated with UF.Methods: A SR according to Cochrane guidelines was completed. A reproducible search strategy was conducted by two independent reviewers in thirteen online databases: the Cochrane Central Register of Controlled Trials, Web of Science, SCOPUS, B-On, SportDiscus, Scielo; APA PsycINFO, Psychology and Behavioral Sciences; Academic Search Complete; Medline (PubMed); ERIC; Google Scholar; Open Acess Thesis and Dissertations. The search occurred from 1st to 30th June 2020, and there were no limitations regarding the year of publication. Original papers that contained relevant data regarding psychological domains in the context of UF in English, Portuguese and Spanish were selected. The combination of the main terms \u201cultimate frisbee\u201d and \u201csport psychology\u201d was used in all databases. A total of 464 studies were identified and selected in the last phase of selection. After the Screening (n = 301) and Eligibility (n = 71) phases, a total of 30 potential papers were selected and classified. Finally, only four papers were qualified to be included in the final version of SR.Results: The psychological dimensions revealed in the present study were: leadership; basic psychological needs; behaviors; task cohesion and performance; intrateam communication; performance-avoidance goals; friendship goals; sportsmanship associated with goal-directed self-talk and self-regulated learning.Discussion: To our knowledge, this is the first SR about UF. In reviewing all the findings in the studies, there is evidence that UF can promote teamwork, task cohesion, leadership, and increase friendship-approach goals.Conclusion: The results revealed that group goals and promoting teamwork significantly predicted social cohesion and that teamwork and task cohesion was mediated by communication. UF is characterized by communication between all players, whether they are from the same team or the opposing team. In summary, the current study revealed real-time information about the game and its rules. This is important because UF is one of the few team sports worldwide that are self-referred by participants.Systematic Review Registration:https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=169294, identifier: CRD42020169294. The first complete description of Ultimate Frisbee (UF), including general and specific rules, equipment, time, scoring, game variations, and other characteristics were presented by Clark et al. . From thTraditionally known as \u201cUltimate\u201d among participants, UF is a fast-paced, non-contact, mixed team sport played with a flying disc or frisbee Griggs, , assemblThe rapid growth of registered UF practitioners in recent years has attracted interest among researchers in the sport sciences and other disciplines. The published literature on UF has generally focused on: (i) physical, cardiovascular and metabolic demands in healthy adults and athletes comCrocket, , 2011.Research conducted by M\u00e9ndez-Gim\u00e9nez et al. found thPsychological factors and their influence in the environment of UF have led researchers and practitioners to observe and take note of the impact of the SOTG during competitive events Robbins, and its To guarantee consistency, accuracy, and replicability in this SR, the following steps were adopted: (i) definition of systematic search terms through the description and operationalization of concepts; (ii) a pilot study of the systematic search of articles to verify the search accuracy in each previously selected database; and (iii) registration of the pre-determined SR protocol in the PROSPERO database, under the number CRD42020169294.a) Ultimate frisbee: Ultimate is a team sport where contact between players is not allowed. It is played by two, seven-person teams, and it can be played with gender-mixed teams. The official field measures 64 meters by 37.57 meters, with 22.86 meters end zones. Each game is played for 48 min and is divided into two 24-min halves Sport and exercise psychology is the scientific study of people and their behaviors in sport and exercise contexts and the real application of that understanding Sport and exercise psychology dimensions: the field of exercise psychology has tended to grow from sport psychology and sport science to become an increasingly important topic in health research and is now associated with areas such as health psychology across thirteen online databases: (i) the Cochrane Central Register of Controlled Trials, (ii) Web of Science, (iii) SCOPUS, (iv) B-On, (v) SportDiscus, (vi) Scielo; (vii) APA PsycINFO, (viii) Psychology and Behavioral Sciences; (ix) Academic Search Complete; (x) Med line (PubMed); (xi) ERIC; (xii) Google Scholar; (xiii) Open Access Thesis and Dissertations. Original articles ; interventional published between 1960 and 2020 investigating the associations between UF and different psychological dimensions were selected. The research procedures were carried out between the 1st to the 30th of June 2020 by the first author, guided by the last author, who coordinated the SR.The initial search was conducted by two researchers who used a list of terms and keywords. The subsequent screening procedures were implemented to determine whether the articles from the initial search were significant for the study. The selected articles in the present QSR met the following selection criteria: (i) original research published in peer-reviewed online international journals indexed in all databases previously identified ; (ii) the articles should contain one or more keywords in the title or abstract to proceed to the screening phase; (iii) reading of the article in full-text and discussion with other experts on the topic. Articles classified as \u201cdistrustful,\u201d but already in the eligibility phase; (iv) were considered articles of open or closed access. In the case of closed access articles, direct contact was made with one of the authors to obtain the full version of the manuscript.The Selected Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) Statement for the organization of this study was respected , which directs the improvement of the systematic search, operating the extraction. In addition, the Strengthening Reporting of Observational Studies in Epidemiology (STROBE) Positioning Statement was used checked the list of articles and discarded irrelevant hits based on title and abstracts. Then, two reviewers (penultimate and last authors) selected, independently, those papers that fulfilled inclusion criteria. Subsequently, the risk of bias was assessed for each study using Cochrane criteria. Any disagreement was resolved by discussion with all authors. During the process of constructing the SR (mainly considered the pilot search phase), it was found that the evidence gathered did not allow us to select a central outcome to proceed to an SR with meta-analysis. At the end of the search, the small number of selected articles corroborated this point.A total of 464 references were identified through the database in the first phase. Out of these, 163 references were excluded after reading the title and abstract, and replication. After applying these initial criteria, a total of 301 articles entered phase two of eligibility. Of these, 232 papers were later excluded for reasons such as \u201cdealt with other similar modalities,\u201d or \u201capproached study dimensions of different nature,\u201d among others. After the full text of articles was assessed, a total of 71 articles remained eligible, 41 of which were excluded, mainly because they used qualitative research methods. In the last phase of Inclusion, all authors decided that only articles that have psychological dimensions would be included in the final SR, considering the previously presented concepts. As a result, 26 studies were excluded at this stage because they presented social or psychosocial approaches that could cause bias in the presentation of results. In total, four studies were included in the final version of SR.n = 367 female; n = 528), from four different countries participated in these four different studies. Different levels of UF players can be observed across the selected studies such as novice players, student players, university players, and team players. We also observed that the participants' age generally varies between 20.77(\u00b12.03) and 24.30(\u00b13.90) years. The experience of regular and deliberate practice of UF with the same captain varies between 1.25 (\u00b11.30) years in the different selected studies. Lastly, we noted that leadership behaviors , the results indicated that the leadership behaviors of fostering acceptance of group goals and promoting teamwork, high-performance expectations, and individual consideration significantly predicted task cohesion UF players as they are related to gender identities, player interaction, sports landscape, and peace culture.Furthermore, the identification and analysis of UF characteristics and environment can be used to enhance players/students and team's performance. The results revealed that group goals and promoting teamwork significantly predicted social cohesion and that teamwork and task cohesion was mediated by communication. UF is characterized by communication between all players, whether they are from the same team or the opposing team. In summary, the current study revealed real-time information about the game and its rules. This is important because UF is one of the few team sports worldwide that are self-referred by participants.Spirit of the Game sheet as the main differentiating factor. Therefore, there is a need to clarify the motivational self-talk and instructional produced better performance than a control condition for a strength task. An exciting avenue for future research would be important to examine and compare the Spirit of the Game with psychological correlates.The current study provided a systematic review of the psychological domains associated with UF. We identified lines of investigation, but none takes a specific approach to self-refereeing and the use of the SOTG game sheet. Finally, we found that group goals and promoting teamwork significantly predicted social cohesion and that teamwork and task cohesion was mediated by communication. In summary, the current study provides real-time knowledge about the game and its rules as they exist in one of the few team sports that is self-refereed. There seems to be a differentiation in the players' awareness of the game using the The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.JA, RR-G, RA, JC, PT, JV-d-S, and GF: conceptualization, investigation, and resources. JA, RR-G, JV-d-S, and GF: data curation and formal analysis. JA and JV-d-S: funding acquisition. JA, RR-G, RA, JV-d-S, and GF: methodology. JA, JV-d-S, and GF: project administration. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Plantae (6094 occurrences) and Chromista (59 occurrences). This paper, in a standardised form, summarises mostly unpublished materials on the biodiversity of lotic ecosystems.The data paper contains the authors\u2019 materials on the diversity of macrophytes, macroscopic plants regardless of their taxonomic position, in rivers and streams of East European Russia and Western Siberia. These data, collected on 247 rivers and 32 streams in 13 administrative regions of the Russian Federation, were provided as an occurrence dataset presented in the form of GBIF-mediated data. The main portion of the data was obtained in water objects of the Vologda Region (5201 occurrences). In addition, occurrences from the Arkhangelsk Region (347 occurrences), Khanty-Mansi Autonomous Okrug (159), Yaroslavl Region (132), Novgorod Region (97), Kostroma Region (41), Republic of Karelia (31), Sverdlovsk Region (29), Komi Republic (28), Orenburg Region (26), Chelyabinsk Region (22), Voronezh Region (22) and Tyumen Region (18) were given. The studies were carried out mainly in the southern and middle taiga and, to a lesser extent, in the northern taiga and the forest-steppe. The analysed watercourses belong to five drainage basins: the Azov Sea, the Baltic Sea, the White Sea, the Caspian Sea and the Kara Sea. The dataset contains materials on the diversity of The paper summarises the data obtained in long-term studies of phytodiversity in a range of rivers and streams of East European Russia and, fragmentarily, Western Siberia. A total of 6153 occurrences were included in the dataset. According to the GBIF taxonomic backbone, the dataset comprises 292 taxa, including 280 lower-rank taxa and 12 taxa identified to the genus level. All the occurrences are published openly through the Global Biodiversity Information Facility (GBIF) for the first time. Most of the data were stored in field diaries and, thus, by adding the data in GBIF, we believe that other researchers could benefit from it. Macrophytes are an essential component of lotic ecosystems, especially in terms of biodiversity , playingThe study summarises the authors\u2019 materials collected in the rivers of the European part of Russia between 2005 and 2021 and Western Siberia in 2021. A small part (5.5%) of the data was published in the research articles , but maiAll the raw data on the biodiversity in rivers and streams are provided in an occurrence dataset .Diversity, distribution, ecology, biology of aquatic and semi-aquatic plants in the European NorthDmitriy A. PhilippovA list of records of macrophytes in rivers and streams of European Russia and Western Siberia is presented. By macrophytes, we understood macroscopic plants, regardless of their taxonomic position and ecological characteristics. Macrophytes include vascular plants, mosses, liverworts and large multicellular algae . We deteField studies were carried out from May to October, mainly during the greatest development of macrophytes (July and August). The composition of the flora of rivers and streams was established during route field studies. We studied all available microhabitats in the channel and coastal parts of water bodies, including those differing in current velocity, sediments, depths and macrophyte canopy development. When studying streams, the route, as a rule, ranged from 50 to 150 m; on rivers, usually from 100 to 1000 m. In the Vologda Region, studies were carried out in all landscapes; several objects per type of landscape were selected depending on the density of the river network. In other regions, studies were carried out along with the study of other wetland ecosystems. In all studied regions, one route per water object was made. In the field, photographs of plants and their communities were taken, floristic lists were compiled and the main hydrochemical parameters were measured . The studies of the macrophyte community composition were conducted both purposefully and along with the studies of other groups of aquatic organisms. Macrophyte samples were collected; they are currently stored and being registered in the Herbaria of Mire Research Group, Papanin Institute for Biology of Inland Waters Russian Academy of Sciences (coded MIRE) and Vologda State University (coded VO). More than 1000 macrophyte specimens were collected in total.The data were collected and identified by scientists from the Papanin Institute for Biology of Inland Waters Russian Academy of Sciences (IBIW RAS). The accuracy of determination of some samples was confirmed by experts from the Institute of Biology of Komi Scientific Centre of the Ural Branch of the Russian Academy of Sciences and Institute of Biology of Karelian Research Centre of the Russian Academy of SciencesResearch problem formulation.Logistic issues resolution, including the choice of routes, water objects, time and duration of work.Field stage: obtaining samples and other original materials on the flora of rivers and streams. In the field, pictures of plants and floristic lists were taken, some species were collected in a herbarium; several hydrochemical parameters were measured using portable devices .Data collection: analysis of samples not identified in the field or verification of the identification data by the experts. The keys by Records list compilation. The dataset field names were chosen according to Darwin Core . GeorefeThe studies were carried out in various parts of European Russian and Western Siberia, mainly in the southern and middle taiga and a lesser number in the northern taiga and the forest-steppe. The analysed watercourses belong to five drainage basins: the Azov Sea, the Baltic Sea, the White Sea, the Caspian Sea and the Kara Sea. The coordinates of the northernmost site were 64.5729N, 43.2959E, the southernmost site 51.8139 N, 39.3836 E, the westernmost site 58.4353N, 33.2828E and the easternmost site 60.8691N, 76.4263E and 12 taxa identified to the genus level.The dataset contains 292 taxa of PLEASE FILL IN TRAIT INFORMATION HERE2005 to 2021OtherThis work is licensed under a Creative Commons Attribution (CC-BY) 4.0 Licence.Data on macrophyte diversity in rivers and streams of the Vologda Region and several other regions of Russia.https://www.gbif.org/dataset/1c52ce65-b940-4bb8-8666-3025e58ef9edhttp://gbif.ru:8080/ipt/resource?r=rivers_and_streams1Data on macrophyte diversity in rivers and streams of the Vologda Region and several other regions of Russia.Darwin Core31Occurrence datasethttps://www.gbif.org/dataset/1c52ce65-b940-4bb8-8666-3025e58ef9ed1.1Plantae and Chromista (59 occurrences: Ochrophyta - 59) diversity. A total of 6153 occurrences (280 lower-rank taxa and 12 taxa identified to the genus level) are included in the dataset.The dataset contains the authors\u2019 materials on macrophyte diversity (macroscopic plants regardless of their taxonomic position) in rivers and streams of East European Russia and Western Siberia. Overall, the dataset contains materials on The data were collected on 247 rivers and 32 streams of East European Russia and Western Siberia and included 6153 occurrences of macrophytes (280 lower-rank taxa and 12 taxa identified to the genus level).These studies have been conducted since 2005. From 8 to 1404 occurrences were registered each year Fig. . The mosThe studies were conducted in 13 administrative regions of Russia came from each given water object. Almost half of all occurrences (48.9%) were registered on 50 rivers - Potamogetoncrispus, Vulnerable (VU) - Batrachiumcircinatum, Carexatherodes, Carexelatasubsp.omskiana (Meinsh.) Jalas [as C.omskiana (Meinsh.) Jalas], Ricciacanaliculata, Near Threatened (NT) - Aegagropilalinnaei, Moliniacoerulea, Tolypellaprolifera, Least Concern (LC) - Carexpseudocyperus, Charavirgata, Equisetumvariegatum, Hygroamblystegiumfluviatile, Nitellasyncarpa, Seneciopaludosussubsp.lanatus [as S.tataricus Less.] and 17 species assigned with the status \u201cbiological control required\u201d: Batrachiumeradicatum, Batrachiumtrichophyllum, Charatomentosa, Eleocharisuniglumis, Humuluslupulus, Hydrocharismorsus-ranae, Irispseudacorus, Nymphaeacandida, Potamogetonberchtoldii, Potamogetonfiliformis, Rumexhydrolapathum, Scapaniasubalpina, Scolochloafestucacea, Sparganiumnatans, Stratiotesaloides, Typhaangustifolia and Utriculariaintermedia.The studies have provided data on the locations of a number of rare macrophyte species. For example, in the Vologda Region, 32 species listed in the Red Data Book of the Vologda Region were reg"} +{"text": "BACKGROUND: Using a large-scale cohort of about 110,000 people established in 45 areas throughout Japan from 1988 through 1990, the study attempted to uncover the joint effects of combined smoking and alcohol intake on esophageal cancer mortality.METHODS: A cohort established from 1988 through 1990 included 46,465 men and 64,327 women aged 40 years and older and younger than 80. The number of female smokers and drinkers was low, and women were excluded from the analysis for that reason. In addition, 308 people with histories of malignant neoplasm, and 3,579 with unclear smoking and drinking data were also excluded, resulting in 42,578 people available for analysis. A follow-up of these individuals was conducted until 1999. Cox proportional hazards model was used for the analysis.RESULTS: The joint effects of number of cigarettes and amount of alcohol consumed per day were compared with non-smokers and non-drinkers or those consuming less than one unit of alcohol per day. An increased synergistic esophageal cancer mortality risk (3.88) for both smoking and drinking was observed for those smoking 20 cigarettes or less per day and drinking one unit of alcohol or more but less than three units per day, with the risk rising (6.30) for those smoking at least 21 cigarettes and drinking at least three units of alcohol per day. Even in non-smokers with increased alcohol consumption, and in non-drinkers or those drinking at most one drink per day with increased smoking, no increased risk was observed.CONCLUSIONS: In this cohort study of a Japanese population, increased esophageal cancer mortality risk was observed only when both factors of alcohol and tobacco intake were present simultaneously. For women, the number of deaths totaled 110,000, the 7th leading cause of death behind breast, lung, gastric, colon, uterine, and ovarian cancers.4-9 and three cohort studies.10-12 In all such studies, a consistent association was observed with smoking and alcohol intake both in Japan and in other countries, and therefore, that contribution is thought to be real. However, though there have been case-control studies on how smoking and alcohol intake interact to contribute to esophageal cancer,8,9 no detailed cohort-study analysis has been conducted. One reason for this is the need for a long-term follow-up of an extensive cohort. This study\u2019s objective was to elucidate which characteristics of smoking and alcohol intake contribute to esophageal cancer mortality, using data from the Japan Collaborative Cohort Study (JACC Study) for Evaluation of Cancer Risk sponsored by the Ministry of Education, Science, Sports and Culture of Japan (Monbusho). Another purpose was to clarify the joint effects of smoking and alcohol intake.Smoking and alcohol drinking as risk factors for esophageal cancer have been clarified by many case-control studiesDetails of the cohort and follow-up procedures have been described elsewhere.sake (Japanese rice wine), shochu (Japanese spirits), beer, whisky, and wine among current drinkers. The daily amount of alcohol consumption was assessed in terms of the conventional alcohol unit (go) of Japanese sake, one unit of which is equivalent to about 22 grams of alcohol.The baseline data were collected utilizing a self-administered questionnaire, which included details about alcohol consumption, smoking history, dietary habits, health conditions, healthy habits, exercise, occupation, educational background, and subjective view of life. Smoking habit was established by asking the subjects whether they were a non-, ex-, or current smoker. Those who were current smokers were asked about the amount of cigarettes consumed per day and age at which smoking started. Packs were calculated by the number of cigarettes smoked divided by 20, and pack-years were calculated as the product of packs per day and the duration of smoking. Alcohol intake was based on the usual yearly intake of 15 from baseline through 1994, and the International Statistical Classification of Diseases and Related Health Problems, Tenth Revision (ICD-10),16 in and after 1995. Death from esophageal cancer was determined by the coding 150.0 through 150.9 for ICD-9 and C15.0 through C15.9 for ICD-10.The date and cause of death were annually or biannually confirmed, with the permission of the director-general of the Prime Minister\u2019s Office . The date of move-out from the study area was also annually verified by the investigator in each area by reviewing population-register sheets of the cohort members. For deceased subjects, the causes of death were identified with underlying causes of death by reviewing all death certificates in each area once a year with permission from the Director-General of the Prime Minister\u2019s Office. The underlying causes of death were coded according to the International Statistical Classification of Diseases, Injuries, and Causes of Death, Ninth Revision (ICD-9),17 was used to estimate the relative risk due to cigarette smoking or alcohol intake adjusted by age and study centers. To test significance, the two-sided Wald\u2019s test was used. All calculations were performed with Statistical Analysis System\u00ae (SAS) software.18Cox proportional hazards modelAge composition and smoking and alcohol intake status at the time of baseline study are shown in Characteristics of the smokers are shown in Characteristics of the drinkers are shown in Hazard ratio of esophageal cancer by smoking status is shown in shochu (3.40), and sake (2.72). For beer and whiskey no significant increase in risk was observed.Hazard ratio of esophageal cancer based on drinking status is shown in Results of investigation into the joint effects of smoking and drinking are shown in 19 However, this study did not observe any association with cumulative amount of cigarette consumption or cumulative alcohol intake. The reason for this disparity is possibly because, compared with France, alcohol consumed in Japan is largely beer, which has low alcohol content, with a low level of wine intake. Regarding cumulative amount of smoking, relatively low consumption of high alcohol content drinks may have made any significant association hard to detect.For the 42,578 males, a follow-up was conducted over a period of about 10 years, analyzing the joint effects of smoking and drinking on esophageal cancer mortality. It was found from the follow-up that combined smoking and drinking, established previously as risk factors for esophageal cancer, were also clearly shown to contribute to the mortality in this cohort. In this study with regard to smoking, a trend of greater esophageal cancer mortality risk was observed the longer the duration of smoking, but no dose-response was observed for age at start of smoking, number of cigarettes smoked daily, or cumulative amount of cigarettes smoked. For alcohol intake, a dose-response association was observed for amount of alcohol consumed per day, but for number of drinking years or cumulative amount of alcohol intake, no dose-response association was observed. In a previous case-control study, for smoking, length of smoking period was most strongly associated, whereas for alcohol intake, average amount of alcohol consumption was most strongly associated, information that conform with the results obtained in this study.sake, an alcohol unique to Japan. In terms of esophageal cancer, no association was observed with low alcohol content beer, but the risk rose with wine, shochu, sake, and whiskey, in that order. Wine had the strongest association. However, whether this was because wine drinkers were more likely to contract esophageal cancer than sake drinkers, or whether the risk increased not because wine drinkers drank wine but because they consumed more per day is an issue that requires future investigation. The limitation of this study is high percentage of unknown categories among drinkers. This might cause an information bias.By type of alcohol, Japan is characterized by widespread consumption of 8,9 but such effect has not yet been proven in a cohort study. Through analysis of the relationship, we investigated whether smoking or alcohol intake contributed to the onset of esophageal cancer independently or whether they are a risk only when present simultaneously. For non-smokers, no esophageal cancer mortality risk was observed even in the smokers who consumed the largest amount of alcohol. Among non-drinkers and those who consumed less than one unit of alcohol per day, no increased risk was observed even when daily tobacco consumption increased from 20 cigarettes or less to 21 cigarettes or more. As for cumulative effects among non-drinkers and drinkers with less than 30 unit-years, no increased risk was observed even when cumulative pack-years of less than 40 increased to 40 or more. A synergetic effect was observed in both analyses by amount per day and cumulative amount, but not in the analysis by smoking and drinking status. These findings imply that amounts of smoking and drinking are important to increase esophageal cancer.With respect to the interaction of smoking and drinking, a synergistic effect was proven in case-control studies conducted to date,These results suggest that simultaneous exposure to both factors is more important than independent exposure to either smoking or drinking in terms of the occurrence of esophageal cancer in Japan. The results show that quitting smoking, curtailing alcohol intake, and reducing the consumption of high alcohol content drinks can be expected to greatly reduce the current rate of esophageal cancer occurrence.The present investigators involved, with the co-authorship of this paper, in the JACC Study and their affiliations are as follows: Dr. Akiko Tamakoshi (present chairman of the study group), Nagoya University Graduate School of Medicine; Dr. Mitsuru Mori, Sapporo Medical University School of Medicine; Dr. Yutaka Motohashi, Akita University School of Medicine; Dr. Ichiro Tsuji, Tohoku University Graduate School of Medicine; Dr. Yosikazu Nakamura, Jichi Medical School; Dr. Hiroyasu Iso, Institute of Community Medicine, University of Tsukuba; Dr. Haruo Mikami, Chiba Cancer Center; Dr. Yutaka Inaba, Juntendo University School of Medicine; Dr. Yoshiharu Hoshiyama, University of Human Arts and Sciences; Dr. Hiroshi Suzuki, Niigata University School of Medicine; Dr. Hiroyuki Shimizu, Gifu University School of Medicine; Dr. Hideaki Toyoshima, Nagoya University Graduate School of Medicine; Dr. Kenji Wakai, Aichi Cancer Center Research Institute; Dr. Shinkan Tokudome, Nagoya City University Graduate School of Medical Sciences; Dr. Yoshinori Ito, Fujita Health University School of Health Sciences; Dr. Shuji Hashimoto, Fujita Health University School of Medicine; Dr. Shogo Kikuchi, Aichi Medical University School of Medicine; Dr. Akio Koizumi, Graduate School of Medicine and Faculty of Medicine, Kyoto University; Dr. Takashi Kawamura, Kyoto University Center for Student Health; Dr. Yoshiyuki Watanabe, Kyoto Prefectural University of Medicine Graduate School of Medical Science; Dr. Tsuneharu Miki, Graduate School of Medical Science, Kyoto Prefectural University of Medicine; Dr. Chigusa Date, Faculty of Human Environmental Sciences, Mukogawa Women\u2019s University ; Dr. Kiyomi Sakata, Wakayama Medical University; Dr. Takayuki Nose, Tottori University Faculty of Medicine; Dr. Norihiko Hayakawa, Research Institute for Radiation Biology and Medicine, Hiroshima University; Dr. Takesumi Yoshimura, Fukuoka Institute of Health and Environmental Sciences; Dr. Akira Shibata, Kurume University School of Medicine; Dr. Naoyuki Okamoto, Kanagawa Cancer Center; Dr. Hideo Shio, Moriyama Municipal Hospital; Dr. Yoshiyuki Ohno, Asahi Rosai Hospital; Dr. Tomoyuki Kitagawa, Cancer Institute of the Japanese Foundation for Cancer Research; Dr. Toshio Kuroki, Gifu University; and Dr. Kazuo Tajima, Aichi Cancer Center Research Institute."} +{"text": "Established in the Fall 2018 and based in Washington, D.C., the Coalition to End Social Isolation & Loneliness brings together dozens of national organizations including consumer groups, community-based organizations, health plans, mental and behavioral health organizations, health care innovators, and many others to lead a multi-stakeholder dialogue to address the crisis of social isolation and loneliness in America. The Coalition focuses on three major areas to achieve this goal: Disseminating research findings, developing and advocating for federal and state legislative and regulatory policy interventions, and leading public awareness events in Washington, D.C. and across the nation. The COVID-19 pandemic has greatly accelerated efforts to engage Congress and the Executive Branch on a range of federal policy priorities, including leveraging and advancing social services and supports, supporting health care delivery to support those who are socially isolated and/or lonely, and advancing federally-funded research initiatives."} +{"text": "Phil. Trans. R. Soc. A379, 20200301. (doi:10.1098/rsta.2020.0301)). The issue is concerned with theoretical, numerical and experimental investigations of nonlinear transport phenomena in heterogeneous and metastable materials of different nature, including biological systems. The papers are devoted to the new effects arising in such systems . State-of-the-art methods of numerical simulations, stochastic analysis, nonlinear physics and experimental studies are presented in the collection of issue papers.This theme issue, in two parts, continues research studies of transport phenomena in complex media published in the first part (Alexandrov & Zubarev 2021 This article is part of the theme issue \u2018Transport phenomena in complex systems (part 2)\u2019. The fThe present issue covers the rapidly developing research area of multicomponent materials with complex disordered internal structures . It covers multiple disciplines, ranging from the non-equilibrium statistical physics of microscopic effects to phenomenological non-equilibrium thermodynamics, condensed matter physics and biomedical applications. To present a complex picture of micro- (nano) and meso-temporal and spatial scales in transport phenomena, a study of various heterogeneous materials is included in this issue. Special attention is paid to developing the scientific background of biomedical applications . Experimental and theoretical studies of non-equilibrium and field-provoked structural and phase transformations are included. Also, we have included articles devoted to noise-induced phenomena in non-equilibrium systems, which essentially change their deterministic behaviour ,18.. 2Transport phenomena in multicomponent complex and disordered systems are widespread in nature and actively met in many industrial and biomedical technologies. The structural transformations and non-equilibrium phenomena in these systems affect and determine their macroscopic properties and behaviour . Studying the nonlinear transport of heat, dissolved impurities and particles in such complex media will make it possible to investigate the internal structure of new generations of materials and establish new laws of matter transport in living systems or systems exposed to external influences . This, in turn, will make it possible to develop a theoretical basis for transport processes in complex non-equilibrium, metastable, disordered or self-organizing systems in non-living and living matter, such as mysterious glass transitions, transitions from order to chaos and back, transport of medicinal compounds in human and animal blood vessels, anomalous and nonlinear intracellular transport, and so on."} +{"text": "Synapse emulation is very important for realizing neuromorphic computing, which could overcome the energy and throughput limitations of today's computing architectures. Recently, a research team led by Guangyu Zhang at the Institute of Physics, Chinese Academy of Sciences (CAS), reported the realization of the synaptic function in memristor based on a vertical structure of graphene/ultrathin-gallium-oxide/Ag see Fig. . The ultet al. Chin Phys B 2019; 28: 017304][Li Q"} +{"text": "Surveillance of antibiotic consumption in the community is of utmost importance to inform and evaluate control strategies. Data on two decades of antibiotic consumption in the community were collected from 30 EU/European Economic Area (EEA) countries. This article reviews temporal trends and the presence of abrupt changes in subgroups of relevance in antimicrobial stewardship.For the period 1997\u20132017, data on yearly antibiotic consumption in the community, aggregated at the level of the active substance, were collected using the WHO ATC classification and expressed in DDD (ATC/DDD index 2019) per 1000 inhabitants per day. We applied a range of non-linear mixed models to assess the presence of changes in the consumption of antibacterials for systemic use (ATC group J01) and eight antibiotic subgroups.For the majority of the studied groups, a country-specific change-point model provided the best fit. Depending on the antibiotic group/subgroup and on the country, change-points were spread out between 2000 and 2013.Due to the heterogeneity in antibiotic consumption in the community across EU/EEA countries, a country-specific change-point model provided the better fit. Given the limitations of this model, our recommendation for the included countries is to carefully interpret the country-specific results presented in this article and to use the tutorial included in this series to conduct their own change-point analysis when evaluating the impact of changes in regulations, public awareness campaigns, and other national interventions to improve antibiotic consumption in the community. Since their discovery, antibiotics have played an important role in the treatment of bacterial infections. Access to effective antibiotics remains of utmost importance in modern healthcare.et al.et al.,In 2001, the European Commission funded the European Surveillance of Antimicrobial Consumption (ESAC) project with the aim of collecting comparable and reliable data on antibiotic consumption in Europe. In 2011, this surveillance activity continued as the European Surveillance of Antimicrobial Consumption Network (ESAC-Net),In this article, we apply a range of models to data collected through ESAC and ESAC-Net on the consumption of antibacterials for systemic use (ATC J01) and of eight subgroups in the community during 1997\u20132017.The methods for collecting and analysing data on antibiotic consumption in the community are described in the introductory article of this series.In this article, we focused on antibacterials for systemic use (ATC J01) and eight specific subgroups: tetracyclines (J01A), \u03b2-lactamase-sensitive penicillins and \u03b2-lactamase-resistant penicillins , cephalosporins , combinations of sulphonamides and trimethoprim (J01EE), macrolides (J01FA), fluoroquinolones (J01MA), penicillins with extended spectrum and combinations of penicillins, including \u03b2-lactamase inhibitors , and nitrofuran derivatives (J01XE). These subgroups were selected because they represent first-line antibiotic treatments or are recommended for use in severe and multidrug-resistant bacterial infections.,In this manuscript, we focused on detecting changes in antibiotic consumption by country, rather than on detecting common changes for the EU/EEA as a whole. Therefore, the statistical analysis deviates slightly from the approach followed in the other articles of this series.Model 1: Mixed model without change-points;Model 2: Mixed model with one common change- point (C1);Model 3: Mixed model with two common change-points (C1 and C2 with C1 0.05). After the intervention, the quality-of-life scores of the two groups were statistically different than those before (P < 0.05), and the quality-of-life score of the intervention group was higher than that of the control group after the intervention, and there was a statistical difference between the two (P < 0.05). The quality-of-life scores of the two groups of patients before and after the nursing intervention were compared and analyzed. There was no significant difference in the quality-of-life scores between the two groups before the intervention (P < 0.05). The proportion of patients with limited and severely limited functional activity of the affected limb in the intervention group was significantly lower than that in the control group (P < 0.05). A comparative analysis was made on the functional recovery degree of affected limbs in different time periods after the intervention between the intervention group and the control group. With the extension of intervention time, the number of affected limbs in the intervention group returning to the normal state increased significantly, and the proportion was significantly higher than that in the control group, and there was a statistical difference between the two groups (P < 0.001). The proportion of patients who were basically satisfied with and dissatisfied with postoperative nursing in the control group was higher than that in the intervention group (P < 0.05). The total satisfaction of postoperative nursing in the intervention group was higher than that in the control group, and there was a statistical difference between them (P < 0.05). According to the statistical analysis of the total satisfaction of the two groups of patients with postoperative nursing based on the intelligent medical system, in the intervention group, 107 cases (65.64%), 44 cases (26.99%), and 12 cases (7.36%) were satisfied, basically satisfied, and dissatisfied, respectively, and there were 151 (92.64%) satisfied cases in total; in the control group, 67 cases (31.02%), 99 cases (45.83%), and 50 cases (23.15%) were satisfied, basically satisfied, and dissatisfied, and there were 166 (76.85%) satisfied cases in total. The satisfaction rate of postoperative nursing care in the intervention group was significantly higher than that in the control group, and there was a significant difference between them . It can be inferred that patients' compliance is higher under the continuous nursing mode based on mobile medical care, which is consistent with the research results of Lin et al. [P < 0.05), and the quality-of-life scores of patients in the intervention group were higher than those in the control group (P < 0.05). This indicates that continuous nursing intervention can effectively help patients improve their quality of life. The traditional nursing model for breast cancer patients after surgery was only the simple guidance before discharge and oral education during reviews, which was affected by many factors such as time, space, financial resources, and manpower. As a result, the nursing effect achieved was often not satisfactory [P < 0.001), and the proportion of basically satisfied and dissatisfied patients in the control group was higher than that in the intervention group (P < 0.05). The total satisfaction of postoperative nursing in the intervention group was higher than that in the control group (P < 0.05) [P < 0.05), and the proportion of patients with limited and severely limited functional activity of the affected limb in the intervention group was significantly lower than that in the control group (P < 0.05) [Exercise compliance mainly evaluates whether the exercise mode and duration of patients are consistent with the nursing plan formulated by medical staff . The resn et al. . Under nn et al. . The ressfactory . With thsfactory . It has < 0.05) , 37. Aft < 0.05) , 39. It < 0.05) \u201342.In this study, the Apriori algorithm is introduced to optimize the mining technology of big data. Then, the intelligent mobile medical communication system is established to evaluate the intervention effect of different nursing methods on patients undergoing breast cancer surgery. It shows that the intelligent mobile medical communication system established in this study can effectively evaluate the intervention effect of different nursing methods on patients undergoing breast cancer surgery. However, there are still some shortcomings in this study. This study only analyzes the compliance, the satisfaction degree, and the mobility function of affected limbs of breast cancer patients. There is no further analysis of patients' complications. In the future, relevant questionnaires will be designed to further investigate patients' complications, and the results will be applied to out-of-hospital continuing nursing of other diseases to expand the application scope. In conclusion, the intelligent mobile medical communication system established in this study has potential application value, which provides a new idea for the evaluation of postoperative intervention effect under big data."} +{"text": "The present cross-cultural study examined the factor structure, measurement invariance, and convergent validity of the Stress-Symptom and Well-Being Scales from the Stress and Coping Questionnaire for Children and Adolescents (SSKJ), originally in German, across gender and for five newly developed language versions: English, French, Russian, Spanish, and Ukrainian.N\u2009=\u20095,227) from Germany, France, Russia, the Dominican Republic, Ukraine, and several English-speaking countries participated in the survey study.Children and adolescents was used for validation.The factorial structure (five factors) was confirmed. In multi-group comparisons, confirmatory factor analyses showed partial metric invariance across the different languages. Regarding gender, results showed scalar invariance for all languages, except for Spanish. Gender differences were shown with girls scoring higher on somatic symptoms, sadness, anxiety , anger (French), and well-being . Correlations with indicators of mental health and behavioral problems demonstrated convergent validity.The SSKJ Stress-Symptom and Well-Being Scales showed psychometric evidence for equivalence across the different languages and gender. Thus, this instrument is a useful tool for cross-cultural research in children and adolescents. Health Behaviour in School-aged Children (HBSC), which surveyed 15-year-old adolescents in 37 countries, on average about 4 in 10 adolescents reported psychosomatic complaints such as headaches or trouble sleeping, indicating stress were mostly satisfying. There was also evidence for convergent validity for the SSKJ Stress-Symptom Scales: increased symptom reporting was associated with decreased health-related quality of life, increased behavioral problems and increased anger-related emotion regulation , psychological symptoms , and well-being . With regard to the development of the scales, previously only somatic and psychological symptoms were assessed . It is unclear, however, whether the SSKJ Stress-Symptom and Well-Being Scales measure in a similar manner across girls and boys or different languages. Therefore, the first aim of the present study was to analyse whether measurement invariance of the SSKJ Stress-Symptom and Well-Being Scales manifests for five newly-developed language versions: English, French, Russian, Spanish, and Ukrainian. For this, the study covers various countries, including Western regions , Southern regions (Dominican Republic) and Eastern regions . The availability of an evaluated stress-symptom measure for children and adolescents from Western and Eastern regions, speaking different languages, is of high importance for cross-country studies on stress and well-being. Based on the original German SSKJ Stress-Symptom and Well-Being Scales , France , Russia , the Dominican Republic , Ukraine , and several English-speaking countries such as Australia, Great Britain, Ireland, and the USA (English sample: n\u2009=\u2009684). Gender and age demographics for the different language subsamples are shown in Chi2 \u2009=\u200960.76, p\u2009<\u2009.001. In terms of age , the language sub-samples differed, F \u2009=\u2009120.31, p\u2009<\u2009.001, Eta2\u2009=\u2009104. Children were youngest in the German and Spanish samples and oldest in the Ukrainian sample.Participants were 5,347 children and adolescents recruited from elementary schools and high schools in various countries. Questionnaires in which items remained unanswered were excluded from the analyses. Thus, the final sample consisted of 5,227 children and adolescents from Germany (n\u2009=\u2009100) collected in 2012, and data from the school in the USA (n\u2009=\u200963), collected in 2014. Informed consent was given by the pupils and their parents prior to the start of the study. Participants completed the questionnaires in their classes. The survey lasted about one school lesson and was generally carried out by a trained supervisor or teacher. Participants did not receive any credits for participation. The study was conducted according to the ethical guidelines of the Declaration of Helsinki. Human participants\u2019 protection was approved by the University of Education Weingarten, Germany (the institution at which the corresponding author was when the study started).Pupils were recruited through public schools from both urban and rural areas in the regions of Baden-Wuerttemberg and North Rhine-Westphalia ; Auvergne-Rhone-Alpes, Normandy, and Paris ; Irkutsk ; Jarabacoa ; the greater Odessa area ; the Sydney area ; Pontypridd and Slough ; the counties Cork, Longford, and Westmeath ; and Charlotte . Most of the data collection took place between 2015 and 2017, with the exception of one French school are reported in The Stress-Symptom and Well-Being Scales from the https://www.sdqinfo.org). However, it was not administered in all English and French schools. For this study, we used the subscale Prosocial Behavior (5 items) and the Total Difficulties Score that covers four subscales of mental health problems . Internal consistencies (Cronbach\u2019s \u03b1) are reported in r\u2009<\u2009.10) to their corresponding scale were excluded. This concerned the Total Difficulties Score of both the Russian version and the Ukrainian version .The Strengths and Difficulties Questionnaire established for the original German SSKJ Stress-Symptom and Well-Being Scales by fitting (a) a model with freely-estimated factor loadings, and (b) a model with factor loadings restricted to be equal between the groups. We evaluated if the model with equal loadings, provided a comparably good fit to the model with group-specific loadings. If metric invariance was established, we further restricted the thresholds of the indicators in additional multi-group analyses in order to test for scalar invariance. These analyses evaluated if the indicator-specific thresholds which determine the observed indicator values can be considered equal between groups. Since the chi-square test statistic and, in consequence, the likelihood-ratio test, are sensitive to sample size, fit indices were used for evaluating the model fit . The application of this five-factor model to the five non-German language versions of the SSKJ Stress-Symptom and Well-Being Scales also provided mixed results: The CFI and TLI showed values of .90 or higher for all but the English language version and .95 or higher for the Spanish and Ukrainian sample. The RMSEA coefficients showed values of .06 or less for three of the five versions , the SRMR showed values of .08 or less for four versions . Ranges in loadings were 0.59\u20132.15 for the English language version, 0.37\u20131.30 for the French version, 0.58\u20131.18 for the Russian version, 0.28\u20131.39 for the Spanish version, and 0.65\u20131.19 for the Ukrainian version. In summary, ranges were smallest for the German, French and Spanish sample, and largest for the English one.The model of the German version of the SSKJ Stress-Symptom and Well-Being Scales with the five factors somatic symptoms, anger, sadness, anxiety, and well-being is shown in r\u2009\u2265\u2009.70) were found between somatic symptoms and emotional symptoms as well as within the emotional stress-symptoms. In general, symptom factors correlated negatively with well-being, and the strength of these associations was smaller than the associations within the symptom factors. Especially for the Ukrainian version and the Russian version, factors correlated highly . A subsequent explorative factor analysis for both language versions separately indicated four factors each with eigenvalues > 1 . Yet, additional fit analyses using reasonable alternative models with 3-factor, 4-factor, and 5-factor solutions yielded no model with superior fit to the proposed 5-factor model of the SSKJ Stress-Symptom and Well-Being Scales. Therefore, to ensure comparability, the five-factor structure was retained for subsequent analyses. In sum, configural invariance (the same five-factor structure across different language versions) can cautiously be assumed for the five additional language versions of the SSKJ Stress-Symptom and Well-Being Scales . Therefore, metric invariance was tested.Regarding the correlational structure (Supplementary Table 2), high correlations . The fit statistics for the invariance tests are reported in Testing for scalar invariance revealed that none of the four models\u2019 changes in fit remained within the admissible range. Therefore, scalar invariance was not supported for the different language versions.Following our second testing strategy, the invariance of the factor loadings was evaluated by one multi-group CFA, using each language sample as one group. The fit statistics for the invariance tests are reported in the bottom rows of rs from .26 to .54 across language versions). The correlations with Prosocial Behavior were low and largely not significant (rs < .20). Well-being was positively associated with Prosocial Behavior (.23 to .44), and was negatively associated with the SDQ Total Difficulties Score (\u2212.24 to \u2212.33).The invariance of the factor loadings between boys and girls was evaluated by six multi-group CFAs (for each language separately). The fit statistics for the invariance tests are reported in As shown in To examine the effects of gender on stress-symptoms and well-being across the five language versions for which scalar invariance was confirmed , for each subscale we conducted a covariance analysis (ANCOVA) with gender as the between-subject factor and age as control variable. Results are reported in The present study aimed to investigate the factorial structure and the measurement invariance of the SSKJ Stress-Symptom and Well-Being Scales across gender and different language versions within children and adolescents from various countries, including Western, Southern, and Eastern regions. Moreover, we studied associations with the SDQ as an indicator of mental health and behavioral problems and gender differences. Overall, the SSKJ Stress-Symptom and Well-Being Scales showed psychometric properties that ranged from acceptable to good. The five-factor structure of the original German version , implying that factor loadings are equal for the different languages and between boys and girls (with one exception each). This means that stress-symptoms and well-being measured with the SSKJ have similar meanings across language versions and gender (except for the English version when compared to the German version and for Spanish-speaking boys and girls). Regarding the separate comparisons between the original German version and the different languages, metric invariance was confirmed for the four comparisons, German-French, German-Russian, German-Spanish, and German-Ukrainian, thus including Western , Southern (the Dominican Republic) and Eastern regions . Scalar invariance (i.e. strong invariance) for the different language versions, however, was not shown. Furthermore, the results supported partial metric invariance when analyzing the six language versions together in a full-fledged comparison. Here, the loading of one item of the subscale Anxiety was non-equivalent across the different languages and had to be set free. When excluding the identified item, however, partial scalar invariance was met. Taken together, interpretation problems are most likely relevant to the English version and overall to the subscale Anxiety of the SSKJ Stress-Symptom and Well-Being Scales. We therefore recommend improving the translation of Item 15 . Until then, in cross-country studies, the subscale Anxiety should be considered very carefully or be used with an alternative, less ambiguous anxiety-related adjective for Item 15. The requirement for valid comparisons of latent mean scores across the different languages was not completely met and can be reported as a limitation of the scales. Similar restrictions were documented for the SDQ , evidence for scalar invariance was shown for these five language versions as well. Thus, within each language mean scores of the SSKJ Stress-Symptom and Well-Being Scales can be meaningfully compared between boys and girls speaking English, German, French, Russian, or Ukrainian. One can only speculate about the non-invariance by gender for the Spanish-speaking sample as previous studies supported measurement invariance across gender for a Spanish stress-response inventory were shown , wherein country-specific subsamples were too small to perform separate analyses. However, the samples were always student samples, the access was always via schools, and participants answered questionnaires in their classes. Nevertheless, especially for the Spanish version and the English version (as the most heterogeneous sample in terms of countries), findings should be replicated in larger country-specific samples. Second, regarding internal consistencies, for the SSKJ Stress-Symptom and Well-Being Scales, values greater than .70 were not consistently achieved. Lower values (.60 to .70) were found in particular for the subscales Somatic Symptoms and Anxiety. Reasons might be that the subscales are overall short, the response format is only 3-points, and especially the subscale Somatic Symptoms is heterogeneous in content. For the German version of the SSKJ Stress-Symptom and Well-Being Scales, however, adequate retest reliabilities (To conclude, the SSKJ Stress-Symptom and Well-Being Scales demonstrated partial metric invariance indicating a basic level of comparability of the scales across six different language versions, including English, French, German, Russian, Spanish, and Ukrainian, as was previously documented for the coping scales of the SSKJ questionnaire (Eschenbeck et al.,"} +{"text": "Hemitheaaestivaria was introduced from Europe to North America, first being detected in British Columbia in 1973. Until 2019, its North American range was limited to a restricted area of the Pacific Northwest. Here, we report on the first records of H.aestivaria for eastern North America from three widely separated urban centres in eastern Canada during 2019-2020.The geometrid moth Hemitheaaestivaria , Pasiphilarectangulata (L.) and Therajuniperata (L.) . Here we H\u00fcbner, 789 . However, H.aestivaria has not expanded into interior British Columbia or Washington east of the Cascade/Coast Range crest since establishment in the early 1970s and its North American distribution has remained restricted to a small area of the Pacific Northwest.The first North American specimens of in 1973 . Ferguso in 1973 , but by Langley . Althoug Langley and it n Langley . The ranH.aestivaria was photographed in the Durham Region of southern Ontario. During the summer of 2020, adults of H.aestivaria were photographed in Toronto, Ontario (June 29), Saint-Augustin-de-Desmaures, Qu\u00e9bec and Halifax, Nova Scotia (14 July - 26 July). These represent the first records for eastern North America , the vast majority of these are for 2018 \u2013 2020. For example, the Moths of Ontario iNaturalist project currently has about 144,000 observations for 2020, an increase of about 70% from the previous year . It is therefore possible that H.aestivaria has gone undetected for a number of years prior to 2020. The synchrony in appearance of such widely-disjunct localities in 2020 is noteworthy and is also perhaps best explained by a substantial increase of iNaturalist observers and observations in 2020.It is unclear if the eastern North American records represent new and temporary introductions or if H.aestivaria in the Toronto, Qu\u00e9bec and Halifax Regions and its absence in relatively well-surveyed adjacent regions, indicates multiple points of introduction, probably via major shipping ports-of-entry along the Atlantic coast and St. Lawrence corridor or through aerial transportation. Indeed, the observations from Toronto, Saint-Augustin-de-Desmaures and Halifax are all located within 30 km of an international airport. Since both larvae and adults are relatively large and short-lived, the most plausible mode of transport is as dormant (overwintering) eggs, which could easily escape detection on fruit tree or ornamental nursery stock. DNA sequencing of eastern populations could shed light on their geogrphic origin.The occurrence of H.aestivaria has been conducted. In Europe, it is widespread, but absent from northern Scandinavia and the Mediterranean lowlands (H.aestivaria is apparently able to tolerate relatively cold winters. Nevertheless, its European distribution, combined with the fact that it has failed to colonise inland British Columbia and Washington beyond very mild coastal regions in almost 50 years after its introduction, suggests that it is unlikely that H.aestivaria will expand into interior eastern North America in the near future. However, the species could become more frequent in near-shore regions along the Atlantic coast, St. Lawrence River and Great Lakes, as suggested by its Fennoscandian distribution. The coming years will provide a clearer picture of the colonisation and expansion potential of H.aestivaria in eastern North America, particularly with the considerable surveillance potential that citizen scientist platforms, such as iNaturalist, provide. Its spread would be facilitated through transport of dormant fruit trees or nursery stock, as is the case for another introduced geometrid (To our knowledge, no study on the climatic niche of lowlands . Skou (1lowlands . Althougeometrid ."} +{"text": "As aging in place increases in popularity, it is important to understand potential negative outcomes related to the trend. For this presentation, the conceptual-theoretical-empirical (C-T-E) scoping review technique was used to organize research on in-home falls of community-dwelling older adults. Research and theory were included from the fields of social gerontology, disability, policy, social justice, medicine, rehabilitation, and housing. While research from these multiple fields overlaps, an overarching conceptual framework for organizing this literature was found to categorize the theories into three main conceptual areas. The three conceptual areas are: intrinsic (related to the person only), extrinsic , and interaction between intrinsic and extrinsic . This conceptual framework shares similarities with work by others in use of the terms intrinsic and extrinsic, and it draws on the larger influence of Bronfenbrenner's socio-ecological model. However, this review extends previous work by providing a framework for organizing the contributions to falls research across multiple disciplines."} +{"text": "This study aimed to clarify the neuropsychiatric symptoms of right-sided predominant semantic dementia (SD-R) by comparing them with those of behavioral variant frontotemporal dementia (bvFTD), left-sided predominant SD (SD-L), and Alzheimer\u2019s disease (AD). This study also aimed to identify clinical factors related to caregiver burden for bvFTD, SD-R, and SD-L.The neuropsychiatric symptoms of 28 patients with bvFTD, 14 patients with SD-R, 24 patients with SD-L, and 43 patients with AD were evaluated using the Neuropsychiatric Inventory (NPI) and the Stereotypy Rating Inventory (SRI). Cognitive function was assessed using the Mini-Mental State Examination (MMSE). Dementia severity was assessed using the Clinical Dementia Rating. Activities of daily living were assessed using the Lawton Instrument Activities of Daily Living (IADL) scale and the Physical Self-Maintenance Scale. We compared the NPI and SRI scores among the four groups using the Kruskal-Wallis test. In addition, clinical factors related to caregiver burden, represented by the Japanese version of the Zarit Burden Interview (J-ZBI), were analyzed using multiple regression analysis in the bvFTD, SD-R, and SD-L groups.The NPI total score and the NPI subscale scores of apathy and disinhibition were significantly higher in the bvFTD group than in the SD-L and AD groups. The SD-R group scores were closer to those of the bvFTD group than the SD-L group. The SRI total score and SRI subscale scores for eating and cooking and speaking were significantly higher in the bvFTD, SD-R, and SD-L groups than in the AD group. The NPI total score was significantly associated with the J-ZBI score in the bvFTD group. The NPI total score and Lawton IADL scale score were independently associated with the J-ZBI score in the SD-R group. Furthermore, the NPI total score and MMSE score were independently associated with the J-ZBI score in the SD-L group.SD-R seemed to be a similar condition to bvFTD rather than SD-L regarding behavioral symptoms. Our results suggest that each frontotemporal dementia subgroup requires different approaches to reduce the caregiver burden. Frontotemporal dementia (FTD) is a neurodegenerative disorder characterized by progressive behavioral disorders and/or language disability caused by atrophy and neuronal loss in the frontal and temporal lobes \u20133. FTD iA possible explanation for this discrepancy is that the effect of the laterality of brain atrophy in SD on neuropsychiatric symptoms has not been fully considered. SD is characterized by asymmetrical atrophy of the anterior temporal lobe . IndividRecently, Ulugut Erkoyun et al. reportedFurthermore, the study by Ulugut Erkoyun et al. did not This study aimed to clarify the characteristics of neuropsychiatric symptoms of SD-R, especially focusing on stereotypic behavior, by comparing them with those of bvFTD, SD-L, and AD. In addition, this study aimed to identify clinical factors related to caregiver burden for bvFTD, SD-R, and SD-L.This retrospective observational study was conducted without intervention and in compliance with national legislation and the Declaration of Helsinki. This study was undertaken after obtaining approval from the Ethics Committee of Osaka University Medical Hospital and Kumamoto University Hospital.This study included consecutive patients with bvFTD and SD who visited the dementia clinic of the Department of Psychiatry of Kumamoto University Hospital between April 2007 and March 2016 or attended the dementia clinic of the Department of Psychiatry of Osaka University Medical Hospital between April 2016 and August 2020. In addition, consecutive patients with AD who visited the dementia clinic of the Department of Psychiatry of Osaka University Medical Hospital between April 2016 and August 2020 were recruited as the control group. All patients were examined comprehensively by an experienced senior neuropsychiatrist (M.I.) and underwent routine laboratory tests, neuroimaging studies such as magnetic resonance imaging (MRI) and single-photon emission computed tomography, and standard neuropsychological examinations. Patients with bvFTD were diagnosed according to international consensus criteria for probable bvFTD . PatientWe collected the patients\u2019 demographic information, including estimated disease onset age, through clinical interviews with their caregivers. We defined disease onset age as the time when the patient\u2019s closest caregiver became aware of their cognitive abnormality or behavioral symptoms. The patients\u2019 behavioral and psychological statuses were assessed using the NPI . The NPICognitive function was assessed using the Mini-Mental State Examination (MMSE) and demep < 0.05. We compared the NPI total score, the SRI total score, and each subscale score of the NPI and SRI among groups using the Kruskal-Wallis test. If significant differences were found, a post hoc Dunn test was used. For multiple comparisons, the statistical threshold was set at p < 0.05/17 = 0.0029. Initially, bringing all SD patients together, each variable was compared among the three groups and then among the four groups dividing SD patients into two groups. Additionally, we conducted a multiple regression analysis to clarify the clinical factors related to caregiver burden in each FTD group . The following variables were entered as independent variables: MMSE score, NPI total score, SRI total score, PSMS score, and Lawton IADL scale score. The best models were derived using a stepwise regression analysis. The significance level was set at p < 0.05. All statistical analyses were performed using SPSS version 25.0 .The demographic and clinical characteristics between groups were analyzed using the Kruskal-Wallis test or the chi-square test, given the skewed distribution of data. The significance level was set at Of the 2503 patients who were diagnosed with dementia or mild cognitive impairment in the two institutes, 28, 14, 24, and 43 patients with bvFTD, SD-R, SD-L, and AD, respectively, met the inclusion criteria and NPI subscale scores of euphoria , apathy , and AMB than the SD-all and AD groups and that the bvFTD group had significantly higher NPI subscale scores of agitation and disinhibition than the AD group. Upon comparing the four groups , the NPI total score and NPI subscale scores of apathy were significantly higher in the bvFTD group than in the SD-L and AD groups. NPI subscale score of disinhibition was significantly higher in the bvFTD group than in the SD-L and AD groups and significantly higher in the SD-R group than in the AD group . NPI subscale score of AMB was significantly higher in the bvFTD group than in the AD group . However, there were no significant differences between the bvFTD and SD-R groups for all NPI items. Upon comparing the three groups , the bvFTD and SD-all groups had a significantly higher SRI total score and SRI subscale scores of eating and cooking , roaming , speaking , and daily rhythm than the AD group. SRI subscale score of movements was significantly higher in the bvFTD group than in the AD group . Upon comparing the four groups , the SRI total score and the SRI subscale score of eating and cooking and speaking were significantly higher in the bvFTD, SD-R, and SD-L groups than in the AD group. The subscale score of roaming was significantly higher in the bvFTD and SD-L groups than in the AD group . The daily rhythm score was significantly higher in the bvFTD and SD-R groups than in the AD group . Figure The NPI and SRI results are shown in Table H = 21.56, p < 0.001) was significantly associated with the J-ZBI score in the bvFTD group and the Lawton IADL scale score were independently associated with the J-ZBI score. In the SD-L group, the NPI total score and MMSE score were independently associated with caregiver burden.The J-ZBI score was significantly higher in the bvFTD group than the SD-L and AD groups and significantly higher in the SD-R group than in the SD-L group , which may have caused a statistical type 2 error. Third, this was a cross-sectional study at the first visit, restricting some of our interpretations. Fourth, it is well known that the age of onset among bvFTD, SD, and AD varies [There are some limitations that need to be addressed. First, the diagnosis relied solely on a clinical basis without histopathologic and genetic confirmation, with some uncertainty about the rate of misclassification. Second, the sample size of the SD-R was relatively small (D varies , 41. In D varies \u201344. TherPatients with SD showed different behavioral profiles depending on the predominantly atrophic side, and SD-R seems to be a similar condition to bvFTD rather than SD-L in terms of behavioral disorders. Caregiver burden for SD-R was comparable to that of bvFTD and higher than that of SD-L patients. Different factors influenced the increased caregiver burden among bvFTD, SD-R, and SD-L. Overall, the results of this study suggest that it is important to distinguish SD into SD-R and SD-L and take different interventional approaches for FTD among bvFTD, SD-R, and SD-L."} +{"text": "Growth in older populations, and hence in the number of persons living with dementia, is particularly rapid for individuals of Mexican origin living in the U.S. and Mexico. In order to identify influences on cognitive health in this diverse population, the University Texas at Austin and Mexican National Institute of Geriatrics (INGER) organized their second Bridging Conference titled: \"Framing Challenges of Cognitive and Mental Health Care in Mexican-origin Older Adults in Mexico and the U.S\". In this presentation, we highlight the results of a consensus-building session, during which bi-national expert opinions were generated and synthesized addressing gaps in research, knowledge, and policy, as well as the setting of priorities for immediate action and future research. Reducing barriers to adequate care for those aging-in-place with dementia was a central theme of the identified priorities. Critical areas of identified need, more specifically, included reducing social isolation, caregiver burden, and diminishing retirement income."} +{"text": "This study aimed to systematically review the association between telomere length (TL) and muscular fitness. In October 2020, an articles search was applied to PubMed, Scopus, and Web of Science. Eligibility criteria included: cross-sectional, prospective, and experimental study design; outcomes included TL; results expressed the relationship between muscular fitness and TL; studies published in English, Portuguese, or Spanish. Nine studies were included in the review. Results from the four prospective studies are mixed. In one study, the changes in TL were associated with grip strength. Another study concluded that longer mid-life TL was associated with increased grip strength later in life. However, in the other two studies, the association between TL and sarcopenia was not strong. Nevertheless, longer TL was associated with a slower decline in grip strength in older people. From the four cross-sectional studies, three indicated that TL was associated with muscular fitness. On the other hand, in a study with powerlifters, TL remained within the range of values found in subjects with no history of regular strength training, supporting the notion that muscular fitness was not associated with TL. The cross-sectional and prospective studies showed that the relationship between TL and muscular fitness is not conclusive. It seems that there is a positive association between TL and muscular fitness in middle-aged and older adults. However, among younger adults, this relationship was not observed. Chromosome ends are protected by tandem repeats of hexanucleotide units named telomeres guidelines for full-text screening and rating of relevance and risk of bias. A third reviewer solved eventual disagreements about the risk of bias and made the final decision. The methodological quality of the included articles was assessed with a total score ranging from zero (lowest) to 11 .The methodological quality of the articles was assessed by two researchers independently using the Physiotherapy Evidence Database (PEDro) scale. Agreement between reviewers was assessed using k statistics can influence the association between TL and muscular fitness and, therefore, may contribute to the variety of observed results. Also, some of the studies included in the review had small sample sizes. Nonetheless, the small number of empirical studies on the topic existing so far does not allow establishing a stratification of evidence by those additional factors. Additionally, the variety of study methodologies and outcome measures makes it impossible to adequately perform a meta-analysis.The existing empirical evidence on the relationship between TL and muscular fitness is mixed and may be influenced by additional factors, such as individuals' age. In this regard, it seems that there is a positive association between TL and muscular fitness in middle-aged and older adults. However, among younger adults, such a relationship may not be evident.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.AM, MP, DH-N, and AI: conceptualization. AM, MP, and DH-N: methodology development. MP and PM: formal analysis. MP and DH-N: investigation. AM, MP, and PM: writing original draft. EG, GF, JM, HS, and AI: writing review and editing. EG, GF, HS, and AI: visualization. HS and AI: supervision. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Yu et al.; Zhou et al.). For instance, S deficiency can limit Fe acquisition . On the other hand, P excess can diminish Zn acquisition . In other cases, a scarce element, i.e., K, can be substituted by another element of similar characteristics, like Na .Plants, like other living organisms, require an assemblage of essential elements to synthesize their constituent compounds and for essential metabolic reactions. Besides carbon (C), hydrogen (H) and oxygen (O), plants require 14 essential mineral elements such as nitrogen (N), phosphorus (P), potassium (K), sulfur (S), magnesium (Mg), calcium (Ca), zinc (Zn), iron (Fe), copper (Cu), manganese (Mn), molybdenum (Mo), nickel (Ni), chlorine (Cl) and boron (B) . The minireview deals with the interaction between N and P in the development of root nodules and cluster roots . Finally, within the 13 original research articles; eight of them are about the interactions between two or three elements, including non-essential ones, such as Fe-Cu, Si-Fe, Fe-P-S, S-N, Fe-Zn, K-Na, or P-Zn ; four of them are about the interactions among many nutrients, such as ionome-macronutrients, ionome-micronutrients, ionome-N ; and one is devoted to the substitution of K by Na .This Research Topic updates recent results showing the interactions between different essential mineral nutrients, and also between essential and non-essential ones. It includes 5 reviews, 1 minireview, 1 perspective and 13 original research articles. Regarding the reviews; one is related to interactions between two essential elements, S and Fe , and also direct interactions, like those studying the interplay between 2 and 3 elements . The depicted interactions occur at different steps of nutrient acquisition and translocation inside the plant. For instance, P deficiency, through organic acid release and rhizosphere acidification, can promote the mobilization of other nutrients, like Fe or Zn . In relation to the translocation of some elements, like Fe, Cu and Mn, S deficiency can negatively affect it by limiting the biosynthesis of nicotianamine, a chelating agent linked to this process . One element can participate in compounds or proteins involved in key processes related to others [i.e., S-containing metabolites participate in the synthesis of ethylene and phytosiderophores, which are in turn implicated in Fe uptake . For instance, ethylene and nitric oxide upregulate both P- and Fe-acquisition genes in such a way that the deficiency of either of them, that stimulate the production of ethylene and nitric oxide, promote the acquisition of the other one .In this Research Topic, mechanisms underlying the observed interactions are proposed. Two elements can interact because they share similar chemical properties, like K and Na . Additionally, non-essential elements, like Si, can affect the homeostasis of essential elements . A better knowledge of such interactions could aid in the improvement of some nutritional disorders and/or in the biofortification of some essential elements for humans and animals, like Se .Interactions between nutrients can have many different consequences, depending on them being essential or beneficial, and on other factors. In this sense, it is important to point out that the interactions between nutrients greatly depend on the severity of the nutrient deficiency or excess could lead to more rational fertilization practices, preventing interactions that could contribute to an unbalanced mineral nutrition of plants. This knowledge is also necessary to obtain more efficient genotypes in the acquisition of the different nutrients.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.PL was funded by the National Natural Science Foundation of China (32070279). FJR and RPV were funded by the Spanish Ministry of Science and Innovation (RTI2018-097935-B-I00), the Spanish State Research Agency, through the Severo Ochoa and Mar\u00eda de Maeztu Program for Centers and Units of Excellence in R&D (Ref. CEX2019-000968-M), and the \u2018Junta de Andaluc\u00eda\u2019 (Research Groups AGR115 and BIO159).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The first Sustainable Development Goals (SDGs) of The United Nations aims to \u201cend poverty in all its forms everywhere\u201d. Its seven associated targets aim, among others, to eradicate extreme poverty for all people everywhere. In Vietnam, poverty eradication in ethnic minorities and mountainous areas are among the top priorities. This study aims to learn about farmers\u2019 livelihoods associated with perceived difficulties in Chau Thai Commune, Nghe An Province, a rural mountainous area in Vietnam. A random sampling technique and a face-to-face interview method were employed to conduct a field survey in the region in 2018. The dataset collected from 215 households shows that Chau Thai Commune's livelihood largely depends on agriculture and forestry. Plantation forest and livestock are major sources of farmers\u2019 income while forestland accounts for over 90% of households\u2019 land. Besides, the disparity in livelihood in areas such as forestland, labor and income between the poor and non-poor households is reported. This primary data could be useful for scholars who want to conduct a further in-depth study and or experts, policymakers who work in Vietnam's \u2018New Rural Development\u2019 program to devise a better rural livelihood -improvement policy for farmers, particularly the poor in the uplands of Vietnam and beyond. Accordingly, forest development has been a high priority in many parts of the world including Vietnam After eliminating some incomplete answers, our data presented in this study includes 215 observations with information on three aspects: (1) resource structure and the local people's livelihood strategies, (2) factors hindering production forest planting, (3) the difference between poor and non-poor households, and (4) the personal information of the head of households. It is noted that households are categorized into poor and non-poor according to Vietnam's poverty line in rural areas between 2016 and 2020 ($1.02 per person/day) Moreover, people in Chau Thai commune faced many difficulties in taking out loans. Despite easy access to loans, the mean values from 3.00 to 3.35/5 on grounds of short loan repayment period, high interest rate and limited amount of money lent. More importantly, most families had no or little capital. Labor is also an essential factor in the quality of economic development of the region. In Chau Thai, generally, the labor force had many limitations in both quantity and quality. On top of production factors, the market for forest products also posed major obstacles for indigenous households. For example, the statement that prices of forest products were unstable and volatile with the reported average score is 3.96/5.Moreover, revenue from plantation forests also mad2Experimental design: We employed the probability sample approach to make inferences beyond the sample in this study. Accordingly, we selected Chau Thai Commune as a study area for data collection for two reasons. First, Chau Thai is one of 206 poor communes in Nghe An Methods: We conducted a survey in 2018 to obtain primary data on farmers\u2019 livelihoods and perceptions of difficulties regarding the dimensions of livelihood, following the methods of To capture the features of the respondents\u2019 livelihood and their perceptions towards the constraints presented in As for this research, there is still room for improvement, and acknowledgement of the limitations allows the research quality to improve The authors declare that this study is designed for the purpose of research and goodwill. Interviews are conducted on the basis of willingness and mutual consent, and participants\u2019 personal information remain confidential.Quan-Hoang Vuong: Conceptualization, Methodology, Formal analysis, Supervision and validation, Writing - Original draft, Writing - Review & editing; Phu Pham: Conceptualization, Methodology, Formal analysis, Writing - Original draft, Writing - Review & editing; My-Hien Nguyen: Formal analysis, Writing - Original draft, Writing - Review & editing; Cong-Thang Ngo: Formal analysis, Visualization, Writing - Original draft, Writing - Review & editing; Phuong-Mai Tran: Formal analysis, Writing - Original draft, Writing - Review & editing; Quy Van Khuc: Conceptualization, Methodology, Data curation, Project Administration, Formal analysis, Supervision and validation, Writing - Original draft, Writing - Review & editing. All authors have read and agreed to the version of the manuscript.The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper."} +{"text": "A recent study concerning the \u201cImpact of COVID-19 on the Interrelation of Physical Activity, Screen Time and Health-Related Quality of Life in Children and Adolescents in Germany\u201d was investigated by Wunsch et al. was usedhttps://www.kidscreen.org/english/questionnaires/kidscreen-52-long-version/, accessed on 22 February 2021).A total of 10 dimensions of HRQoL are considered in the basic KIDSCREE\u2018N 52: physical well-being, psychological well-being, moods and emotions, self-perception, autonomy, parent relations and home life, social support and peers, school environment, social acceptance (bullying), and financial resources. KIDSCREEN-27 and KIDSCREEN-10 consist of five dimensions thereof sensations associated with food intake, among them sensory perceptions such as taste and smell, and increasing feelings of satiety. As is well-known, presentation and visual impact also play an important role in the pleasure of eating, especially for children, and not least the social environment of eating with the family or with friends at school.The burning glass nature of the Coronavirus pandemic may be an opportunity to re-evaluate the contribution of nutrition to the health-related quality of life in children."} +{"text": "The Rush Caregiver Health and Well-Being Initiative (Caregiver Initiative) draws together evidence-based practices into a single framework to improve care for older adults and caregivers. The Caregiver Initiative has two components: system-level and caregiver level interventions. The complexities of system change take place within leadership, data management, and provider teams throughout the health care system, and solutions to resistance have been developed. Caregiver-level interventions start with an assessment using evidence-based tools, and offer an opportunity to participate in a Teach-Back Clinic, Family Care Planning sessions, and/or Goals of Medical Care meetings, all connected to the 4Ms of an Age-Friendly Health System. Contact and follow-up issues were addressed, and as of February 2021, 191 caregivers have enrolled. Outcomes to date show statistically and clinically significant reductions in depression, anxiety, and caregiver burden. This presentation will highlight lessons learned in the development of the model and caregiver outcomes to date."} +{"text": "The Republic of Korea has a high incidence of tuberculosis (TB) and TB-specific mortality rate. In 2019, it had the second highest TB-specific mortality among Organization for Economic Co-operation and Development countries. Understanding the factors associated with TB-specific deaths may help eradicate the disease. Therefore, we aimed to identify the general characteristics associated with TB-specific mortality among Koreans. Using Causes of Death Statistics data from Statistics Korea, we assessed the year of death, sex, age, occupation, area of residence, marital status, and education level reported between 2008 and 2017. Patient characteristics associated with TB-specific deaths were analyzed using the Chi-squared test, while influencing factors of TB-specific mortality were analyzed using logistic regression analysis to calculate adjusted odds ratios (AOR). Female , those with a graduate degree or higher had lower TB-specific mortality rates than those of their counterparts. Conversely, those aged \u226570 years , single , and skilled agricultural, forestry, and fishery workers had higher TB-specific mortality rates than those of their counterparts. In conclusion, TB-specific mortality rates differed according to the characteristics of the deceased patients. In order to establish effective TB control, multisectoral action on broader determinants should be strengthened. Mycobacterium tuberculosis (MTB) (Tuberculosis (TB) is an airborne infectious disease caused by is (MTB) . MTB hasis (MTB) . Approxiis (MTB) , 4. MTB is (MTB) , 5\u20137. Reis (MTB) .Since 1993, TB has been considered a global public health emergency relative to 2015 . The KCD-7 is a set of disease classification codes that were modified from the International Classification of Disease 10th revision (ICD-10) by WHO to reflect the situation in the Republic of Korea. In the present study, TB-specific mortality was defined as cause of death corresponding to KCD-7 code A15 , A16 , A17 (tuberculosis of nervous system), A18 (tuberculosis of other organs), or A19 (military tuberculosis).Among the 2,647,823 deaths reported between 2008 and 2017, cases with missing data and errors were excluded, resulting in a total of 2,589,557 deaths included in the analysis set. Of the 2,589,557 deaths in the analysis set, there were 21,968 deaths by TB and 2,567,589 deaths by other causes .The time of death, sex, age, occupation, area of residence, marital status, education level, and place of death were considered in the analysis. TB deaths in Korea were declining between 2008 and 2017 , and yeaChi-squared test and logistic regression analyses were used for statistical analysis. The Chi-squared test was performed to identify the characteristics of deceased patients that were associated with TB-related deaths, while a logistic regression model was established to analyze the characteristics of the deceased patients that influenced TB-related deaths. The logistic regression model was established by setting death due to TB as the outcome variable and year of death, sex, age, occupation, area of residence, marital status, and education level as the explanatory variables. SAS 9.4 was used for all statistical analyses and the significance level for all statistical testing was set to 5%.The study was conducted with approval from the Korea University Medicine Institutional Review Board (IRB No. 2019AS0245).The Chi-squared test was performed to analyze the associations between death from TB and deceased patient characteristics . All chan = 14,049) and those aged 75\u201384 years . Statistical differences were found in gender and age distribution with non-tuberculosis deaths. Regarding TB-specific deaths according to occupation, the most frequent deaths were among students, homemakers, and unemployed subjects , followed by skilled agricultural, forestry, and fishery workers , and other occupations . In non-TB-specific deaths, the most frequent deaths were also among students, homemakers, and unemployed subjects . Moreover, the percentage of deaths from TB was high among non-metropolitan area residents and among singles , while a decreasing trend in the percentage of deaths from TB was observed among patients with a higher education level. Finally, regarding the place of death, the percentage of in-hospital deaths from TB was higher than that of out-of-hospital deaths by TB .The percentage of deaths from TB, according to the year of death, showed a decreasing trend. However, deaths from TB, according to the month of death, showed a similar distribution pattern, indicating no association. Regarding TB-specific deaths according to sex and age, the most frequent deaths were among male patients . In EuroThe present study had some limitations. First, the data used in this study were complete enumeration survey data that were collected for establishing national policies and not for research purposes. However, the data were highly representative of all deaths. Therefore, the data were meaningful in that the characteristics of the target population could be estimated with minimal selection bias. Second, because only patients who died from TB were analyzed, the findings cannot be generalized for all TB patients. However, the present study was significant in that it differentiated deaths by TB from deaths by other causes. Lastly, the data used in the analysis were not adjusted for factors that have a definitive influence on TB-specific mortality, such as personal nutritional status, smoking status, diabetes, and HIV status . DespiteDespite such limitations, the present study provides extensive epidemiological analysis data over 10 years on the TB-specific mortality rate in a country with a high burden of TB and identifies factors associated with TB-specific deaths and differences between characteristics. Moreover, the study presents aspects of TB control policies in the Republic of Korea that should be updated based on the findings of the study.The present study used Causes of Death Statistics data from Statistics Korea to describe the percentage of TB-specific deaths reported between 2008 and 2017 in the Republic of Korea according to patient characteristics. This study also analyzed the factors associated with the TB-specific mortality to present aspects of TB control policies in the Republic of Korea that should be upgraded to reduce the mortality rate. Regarding the characteristics associated with the TB-specific mortality, year of death, sex, age, occupation, region of residence, marital status, education level, and place of death were identified as factors that may influence the mortality rate, but season did not show a significant association with TB-specific mortality. Accordingly, based on the findings of the present study, it is necessary to actively consider the positive aspects of TB control policies in other countries to use as a guide for implementing appropriate policies to eradicate TB in the Republic of Korea.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding authors. The raw data can be found at the Statistics Korea.The present study was conducted with approval from the Korea University Medicine Institutional Review Board (IRB No. 2019AS0245). Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.SC and J-YS: writing\u2014original draft, conceptualization, data curation, formal analysis, validation, visualization, investigation, methodology, and writing\u2014review & editing. I-HO: supervision, writing\u2014review & editing, writing\u2014original draft, and resources. SL: supervision, writing\u2014review & editing, and resources. H-YK: supervision. YL: resources. KB: methodology. All authors contributed to the article and approved the submitted version.This research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (Grant No. HI20C1068).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Along with capital and human resources, an organization\u2019s data represents one of its fundamental assets. Data administration (DA) attempts the effective planning, organization, and management of an enterprise\u2019s data resource, with the intention of empowering the organization to achieve its mission and goals.Achieving enterprise integration, and developing the supporting information technology infrastructure, is a critical need for organizations. Yet despite substantial attention by business managers, technologists, and vendors of tools and methodologies, there are few obvious solutions or guidance on how to accomplish this.The Data Administration Management Association (DAMA) is the professional organization for data administrators. An international board over-sees a loose federation of local chapters in the United States, Canada, Australia, and Europe. The National Capital Region Chapter (NCR DAMA) has monthly meetings from September through April, as well as a Symposium in May.Enterprise Integration in the Turbulent 90s. Attended by over 200 Federal and private industry data administrators, the Symposium was cosponsored by NIST and NCR DAMA.NCR DAMA held its seventh annual Symposium at NIST on May 17\u201318, 1994. The theme this year was The Symposium emphasized the practices, technologies, activities, initiatives and ideas that deliver clearly visible value to the users, or \u201ccustomers\u201d of data administration. In addition to presentations by nationally recognized experts and practitioners, it included breakout dialog sessions and panel discussions. Topics ranged from the keynote speech on the National Information Infrastructure to the latest implementation of the Information Resource Dictionary System (IRDS) standard. New this year was a Vendor Exhibit Area, where the latest tools for implementing the practices of Data Administration could been seen.The keynote speaker was Arati Prabhakar, Director of NIST, describing the government\u2019s role in the National Information Infrastructure.Mary \u201cBunny\u201d Smith provided a personalized perspective of information systems design for new, very small businesses. Spreadsheets are invaluable both for planning and cost accounting. One can not start to model one\u2019s business too soon, even if it occurs on the kitchen table.Dr. Mike Mestrovitch, DoD, discussed transforming the enterprise through Enterprise Integration, and what transformation is planned for DoD. It will involve changing every aspect of the organization to meet new circumstances and expectations. It is a way of using information as a strategic asset to manage the enterprise far more effectively and efficiently. It bridges functional and technical boundaries to increase flexibility and to focus all available capabilities on mission results. The process includes: establishing a vision for the future; creating a sense of urgency about the vision; redefining the business processes; redefining resource capabilities; creating a new work environment; re-creating management systems and structures; and building an information and technology architecture to empower the organization to execute.The last speaker, John Zachman of Zachman International, presented his Information Architecture concept together with his personal view of the future of manufacturing and technology. We must change from Custom-design-and-build and Provide- from-stock to Assemble-to-order processes. He issued a challenge for everyone in the audience to rise to meet \u201cThe New Realities of the Information Age.\u201dA series of concurrent sessions focused on various managerial and technical aspects of enterprise integration. Several sessions addressed the National Performance Review. Others considered data modeling, EDI, and open systems.Repository, A Missing Link in Enterprise Integration, Carla von Bernewitz, DoD, moderator;Using IDEF in Conjunction with Information Engineering, Vince Cordovano, James Martin Consulting, moderator;Standards for Data Administration, Judith J. Newton, NIST, moderator;GOV-SIG: Stepping Stone to Public Sector Integration, Pam Piper, DoD, and Tom Kurihara, NIST, moderators;Linking IRM Initiatives to Strategic Planning, Stephanie Wietecha, FC Business Systems, moderator;Reengineering a Data Management Program, Patricia Simes, SRA, moderator.The following panel discussions were presented:The proceedings of this, the Third, the Fifth, and the Sixth Symposia were distributed at the event. The Proceedings of the First,"} +{"text": "Four major international collaboration clusters were found in MPP-AD research. The country collaboration network in MPP-AD was characteristic of sparse interaction and acquaintanceship . Overall, international collaboration is globally inadequate against the robust authors' collaboration index of 5.71 in MPP-AD research. Furthermore, four conceptual thematic frameworks (CTF) namely, CTF#1, roles of microbial/microbiome infection and dysbiosis in cognitive dysfunctions; CTF#2, bacterial infection specific roles in dementia; CTF#3, the use of yeast as a model system for studying MPP-AD and remediation therapy; and CFT#4, flow cytometry elucidation of amyloid-beta and aggregation in Saccharomyces cerevisiae model. Finally, aetiology-based mechanisms of MPP-AD, namely, gut microbiota, bacterial infection, and viral infection, were comprehensively discussed. This study provides an overview of MPP-AD and serves as a stepping stone for future preparedness in MPP-AD-related research.Microbial infections have been linked to the pathogenesis and pathophysiology of Alzheimer's disease (AD) and other neurodegenerative diseases. The present study aimed to synthesise and assess global evidence of microbial pathogenesis and pathophysiology in AD (MPP-AD) and associated neurodegenerative conditions using integrated science mapping and content analytics to explore the associated research landscape. Relevant MPP-AD documents were retrieved from Web of Science and Scopus according to PRISMA principles and analysed for productivity/trend linked to authors/countries, thematic conceptual framework, and international collaborative networks. A total of 258 documents published from 136 sources to 39.42 average citations/document were obtained on MPP-AD. The co-authors per document were 7.6, and the collaboration index was 5.71. The annual research outputs increased tremendously in the last 6 years from 2014 to 2019, accounting for 66% compared with records in the early years from 1982 to 1990 (16%). The USA ( Alzheimer's disease (AD) is a progressive neurological condition associated with degeneration of neurons, memory loss, learning impairment, and significant changes in character and behavioural activities and AD has been established , BibTeX (bib), and Tab-delimited formats for pre-processing and further analyses.For the systematic assessment of microbial pathogenesis and pathophysiology of Alzheimer diseases (MPP-AD), we searched the Web of Science (WoS) and Scopus for relevant studies using the guidelines of the \u201cPreferred Reporting Items for Systematic Reviews and Meta-Analyses\u201d based factor analysis , articles in book chapters (n = 9), and articles in proceedings papers (n = 9). A total of 1,391 prolific researchers authored these documents, with about 15 authors of single-authored documents and 1,376 authors of multi-authored documents, which means that multiple authors predominantly wrote the documents. The average citations per document were 39.42, and the documents consist of 1,692 keywords plus and 579 author's keywords. The documents per author was 0.185, and the authors per document were 5.39. The co-authors per document were 7.6, the collaboration index was 5.71, and this value indicated a high collaboration among the researchers that authored these documents.The results of the annual research outputs in the MPP-AD research landscape were represented in n = 12 articles and 563 citations, h-index 11) and publication start year from 2005. Macreadie I had 10 articles with 169 citations, h-index 6 from 2008, whereas Alonso R published nine articles with 329 citations with h-index 8 from 2014. Ball M started publishing in 1997 on the subject, and he had nine articles, 133 citations with h-index 5. According to Grech and Rizk , PLoS ONE , Handbook of Infection and Alzheimer's Disease (seven articles and 10 citations), Frontiers in Ageing Neuroscience (six articles and 292 citations) and Neurobiology of Ageing (six articles and 232 citations). The quality of a scientific journal is usually measured by its impact factor, and we observed that Alzheimer's research has received reputable attention in high impact factor journals. It is also remarkable to note that the first top journal was dedicated to Alzheimer's research. This is a multidisciplinary journal to facilitate understanding the aetiology, pathogenesis, and treatment of Alzheimer's disease. It has an impact factor of 3.909 and well-indexed in several scientific databases.The most relevant journals on MPP-AD are presented in This section describes the collaboration network between the countries involved in MPP-AD research, and the results are represented in International collaboration clusters range between clusters of 2\u201310 countries. Four major collaboration clusters were found. The 2-nation purple cluster depicts collaboration between Pakistan and Poland. The 4-nation green cluster consisted of a collaboration involving Latvia, Belgium, Switzerland, and France, the 4-nation red cluster comprised scientific networks among Spain, Italy, Hungary, and Japan, and the 10-nation blue cluster agglomerate of Austria, China, Canada, Slovakia, South Africa, Sweden, Germany, USA, United Kingdom, and the Netherlands. Isolated countries on the map show that they lack international collaboration in the MPP-AD research landscape.It could be served that the USA has the largest circle size with the highest collaboration with other countries. It has a direct link with the United Kingdom, Germany, Austria, Netherlands, Canada, China, Sweden, South Africa, Switzerland, Belgium, Spain, Hungary, Japan, Greece, Australia, and Israel. Similarly, the United Kingdom has a collaboration with the USA, Germany, Austria, Slovakia, Netherlands, Canada, China, France and Latvia. Some countries are not connected to the world leaders on MPP-AD research. Scientific evidence has highlighted that research collaboration results in an increase in research outputs, division of labour, exchange of ideas and skills, and research funding . The country collaboration map's density and diameter were 0.11 and 4, characteristic of sparse global interaction, however, with acquaintanceship strength. Overall, the network transitivity and average path length was 53.5% and 2.053, respectively. The centralisation statistics, including degree (40.5%), closeness (4%), betweenness (23%), and eigenvector (76.7%), reveal a global inadequate international collaboration against the robust author collaboration index of 5.71 in the MPP-AD research landscape. For instance, eigenvector and centralisation reveal that 76.7% of nations exhibit independence or lack collaboration with other countries in the MPP-AD research landscape.There is no doubt that global research on MPP-AD has increased enormously over a few decades. A progressive growth was witnessed with the substantial weight of scientific evidence in the volume of published articles on the subject study. Our findings collaborate with the report of Dong et al. . It is qEscherichia coli) endotoxin potentiates beta-amyloid fibrillogenesis in AD and other forms of dementia is centred on the use/development of yeast as a model system for studying AD pathophysiology and remediation therapy based on amyloid-beta and autophagy. The fourth is centred on flow cytometry elucidation of amyloid-beta formation and aggregation in Pichia pastoris and S. cerevisiae) have been beneficial systems in understanding the pathogenesis and pathophysiology of AD and some neurodegenerative diseases. For instance, elucidation of in vivo amyloid beta-mediated aggregation has been demonstrated in yeast and tauopathy in the central nervous system (Hudec et al., Recent reports have shown that gastrointestinal bacterial may play a contributory role in the development of AD (Bostanciklioglu, C. pneumoniae in the mouse. Furthermore, exposure of spirochetes to neuronal and glial cells also showed amyloid-like pathology, which could be linked to neurodegeneration.The gut bacteria also release inflammatory activators, beta-amyloid protein, and other neurotoxic substances that can disrupt the host immune system (Angelucci et al., Viruses, especially the Herpes simplex virus and cytomegalovirus, has been implicated in AD (Cheon et al., The present study reveals a comprehensive science mapping of global research on MPP-AD. We observed an increase in annual productivity in MPP-AD across the study years. Itzhaki R and Lukiw W were the most prolific authors. The USA and the United Kingdom were the most relevant countries with the highest published articles and citations. We observed that the most powerful nations of the world are interested in MPP-AD and AD research. In addition, the Journal of Alzheimer's Disease and PLoS ONE were the most relevant journal on the subject. We also observed a high collaboration network between the researchers, although the research outputs were predominated by single country publication. Findings from this study would be valuable for younger researchers interested in joining the field in identifying potential collaborators. The strength of the study is shown in the first integrated content analysis and systematic science mapping of microbial roles in pathogenesis and pathophysiology of AD. The science mapping and the content analysis also offers a comprehensive overview of the MPP-AD research landscape.However, the shortcoming of the science mapping consists of the use of WoS and Scopus, which might exclude documents in other databases that are not linked to the two databases. In addition, the focus of the analysis is on articles written in English and excluded those written in other languages. Furthermore, the analysis only measured the number of research articles published on the subject without considering the scientific quality of each article.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.TE, TO, and KO designed the review and wrote the manuscript. TE searched the literature. All authors read and edited the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Objectives: This study aimed to investigate the effect of nurse-led, goal-directed lung physiotherapy (GDLPT) on the prognosis of older patients with sepsis caused by pneumonia in the intensive care unit.Methods: We conducted a prospective, two-phase (before-and-after) study over 3 years called the GDLPT study. All patients received standard lung therapy for sepsis caused by pneumonia and patients in phase 2 also received GDLPT. In this study, 253 older patients (age \u2265 65 years) with sepsis and pneumonia were retrospectively analyzed. The main outcome was 28 day mortality.Results: Among 742 patients with sepsis, 253 older patients with pneumonia were divided into the control group and the treatment group. Patients in the treatment group had a significantly shorter duration of mechanical ventilation , and a lower risk of intensive care unit (ICU) mortality and 28 day mortality compared with those in the control group. GDLPT was an independent risk factor for 28 day mortality .Conclusions: Nurse-led GDLPT shortens the duration of mechanical ventilation, decreases ICU and 28-day mortality, and improves the prognosis of older patients with sepsis and pneumonia in the ICU. Aging of the population is a critical worldwide trend. The proportion of individuals older than 60 years has tripled over the last 50 years and will triple again before 2050. This aging has major consequences on the health system, including the intensive care unit (ICU). In the USA, almost half 48.7%) of the patients admitted to an ICU are aged 65 years or older, and patients aged 85 years or older account for 7 to 25% of the admission rate , 2. The 8.7% of tBecause of age-related changes in the body, comorbidity, and malnutrition, the onset of pneumonia is insidious, rapid, and critical, and clinical treatment is difficult in older patients , 7, 8. Lchictr.org.cn (identifier: ChiCTR-ROC-17010750).The GDLPT study was a prospective, two-phase (before-and-after) study conducted in Peking Union Medical College Hospital of China from January 2017 to January 2020. Details of this study, including the inclusion and exclusion criteria, have been published previously . The stuThe protocols for pneumonia in the pre- and post-protocol groups are shown in Baseline data of the patients, including sex, age, underlying diseases, the Sequential Organ Failure Assessment (SOFA) score, the Acute Physiology and Chronic Health Evaluation (APACHE) II score, and the Clinical Pulmonary Infection Score (CPIS), were analyzed. Vital signs, laboratory parameters, arterial blood gases, ventilatory parameters, life-sustaining treatments, and infection-related data during admission were also included. The data used in this study were the worst values in the first 24 h after ICU admission. The primary outcomes of this study were 28 day mortality. The secondary outcomes were the duration of ventilation, the length of the ICU stay, and the ICU mortality rate.t test or one-way analysis of variance for continuous variables. Logistic regression analysis was performed with 28 day mortality as the dependent factor, which was significant (p < 0.2) in univariate analysis.Data were analyzed by IBM SPSS version 21.0 . The frequency and percentage and mean and standard deviation were calculated for descriptive statistics. Bivariate analysis was performed using the chi-square test for categorical variables and the Among 742 patients with sepsis, 253 patients who were older than 65 years were included in this study. P = 0.045], and a lower ICU mortality and 28 day mortality compared with those in the control group.There was no significant difference in the ICU duration between the groups. Patients in the treatment group had a significantly shorter duration of mechanical ventilation , heart failure , tumor , need for renal replacement therapy , and mechanical ventilation days . GDLPT was a protective factor for 28 day mortality , 0.379; 95% confidence interval (CI), 0.187\u20130.766; ortality .Pneumonia is frequently encountered in older patients admitted to the ICU, with an incidence rate of >60% in those with sepsis , 13, 19.With aging, there are significant changes in the anatomical and physiological function of the lungs. Older people are more likely to develop pulmonary complications and underlying diseases, including COPD, aspiration, pneumonia, tumor, heart failure, chronic renal failure, and have a poor prognosis . The GDLIn this nurse-led GDLPT study, pain assessment, delirium assessment, and active early activities were carried out every 6 h by nurses. Delirium varies from 11 to 42% in older patients and has adverse outcomes and high health care costs . Up to 6In this study, nurse-led GDLPT significantly shortened the duration of ventilation. Prolonged mechanical ventilation might result in an increased incidence of ventilator-associated pneumonia, which is clinically meaningful, especially for older patients with pneumonia. A high frequency of antibiotic use and antibiotic resistance in patients is caused by ventilator-associated pneumonia. The prevalence of multidrug resistance is increasing, and ventilator-associated pneumonia caused by multidrug resistance is often fatal in the ICU . SeveralAt present, older patients with a critical illness are frequently admitted in the ICU. However, data on older patients with pneumonia are relatively rare, and there have been few evidence-based medical reports . This stManaging pneumonia in critically ill older patients is a complex issue. Aging, comorbidities, frailty, and other factors in older patients significantly increase the management requirements and risks for those with critical illness. Nurse-led GDLPT significantly shortens the duration of ventilation and improves the 28 day mortality rate in older patients with sepsis and pneumonia. Nurse-led GDLPT is a new clinical intervention for the refined management of older patients with pneumonia, and it promotes the recovery of older patients with severe pneumonia.The original contributions presented in the study are included in the article/chictr.org.cn (identifier: ChiCTR1900025850). The patients/participants provided their written informed consent to participate in this study.The studies involving human participants were reviewed and approved by the Institutional Review Board of Peking Union Medical College Hospital (No. JS-1170). The study was registered at NC, QL, HW, and JS contributed to the conception of the study, data interpretation, and drafted the manuscript. WH, HL, and MZ contributed to the data collection and data analysis. WC and ZL contributed to data collection and interpretation and critically revised the manuscript for important intellectual content. All authors approved the final version of the manuscript.The work was supported by National Natural Science Foundation of China (No. 82072226), Beijing Municipal Science and Technology Commission (No. Z201100005520049), Fundamental Research Funds for the Central Universities (No. 3332021018), Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences (No. 2019XK320040), Tibet Natural Science Foundation (No. XZ2019ZR-ZY12(Z)), and Excellence Program of Key Clinical Specialty of Beijing in 2020 (No. ZK128001).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Center for Innovation in Point-of-Care Technologies for HIV/AIDS at Northwestern University (C-THAN) is a partner in the Point-of-Care Technologies Research Network (POCTRN) of the National Institutes of Biomedical Imaging and Bioengineering. POCTRN\u2019s mission is to drive the development of appropriate point-of-care (POC) diagnostic technologies through collaboration that merges scientific and technological capabilities with clinical need. C-THAN develops POC technologies for improved management of HIV/AIDS in low- and middle-income countries with a focus on sub-Saharan Africa. C-THAN incorporates clinical and user needs with technology expertise and resources to address commercialization and implementation barriers through: 1) assessment of unmet clinical needs in POC testing for HIV/AIDS and its comorbidities; 2) collaborations with physicians, researchers and engineers; 3) development of technical, clinical, industrial and regulatory partnerships; 4) clinical testing of prototype devices; and 5) creation of training opportunities for technology developers, evaluators, and other stakeholders. Technologies supported include tests for detection and monitoring of HIV/AIDS and its common comorbidities including tuberculosis, non-tuberculous mycobacteria, viral hepatitis and HIV-related malignancies. CTHAN relies on collaborations established by Northwestern University in Nigeria, South Africa, Mali and Tanzania, to have impact on the prevention and clinical management of HIV/AIDS.The The use of point-of-care (POC) diagnostics brings testing to or near the site of patient care, allowing immediate triage and treatment or discharge leading to improved clinical and/or economic outcomes . Accurathttps://cimit.org/ja/web/poctrn/home) to build multidisciplinary partnerships and expertise in the development of POC testing to address unmet clinical needs (Center for Innovation in Point-of-Care Technologies for HIV/AIDS at Northwestern University (C-THAN). C-THAN is funded by NIBIB, the Fogarty International Center, and the Office of AIDS Research to develop POC technologies critical for improved management of individuals with HIV/AIDS and to facilitate technology commercialization in LMICs with specific emphasis on sub-Saharan Africa.In 2006, the National Institute of Biomedical Imaging and Bioengineering (NIBIB) created the Point-of-Care Technologies Research Network (POCTRN) , making these the leading causes of mortality due to infectious diseases worldwide . The majC-THAN will support the development of such technologies, facilitate their translation into high quality clinical applications, and importantly promote their production and commercialization in LMICs. To create C-THAN, we have assembled a scientific consortium of clinical and biomedical engineering expertise with a 17-year history of collaboration in addressing infectious diseases in sub-Saharan Africa which includes Northwestern University, Muhumbili University, Stellenbosch University, University of Bamako, University of Cape Town, University of Ibadan, University of Jos, and University of Lagos. We have designed a logo for C-THAN to repreC-THAN\u2019s resources are designed to bridge the gap between biomedical engineering and global health, supporting technological innovation for improved health and healthcare, with an emphasis on developing settings. C-THAN is dedicated to building partnerships between United States and LMIC investigators and prioritizes collaborations with LMIC technology developers.in vitro diagnostic tests having received World Health Organization prequalification (POC rapid diagnostic tests (RDTs) primarily using lateral flow assays to detect HIV antibodies have been in use in LMICs for nearly 20 years, with 27 commercial fication . They hafication ; this haTesting POC technologies (POCTs) in research settings has overestimated the accuracy of diagnostic tests and does not necessarily consider environmental conditions specific to low resource settings . For a tIn addition to HIV serology, other tests that are essential for treatment initiation and monitoring and drug sensitivity testing are not yet readily available at the POC. These tests are critical for identifying infections earlier, preventing onward transmissions, and thus reducing HIV incidence at a population level. To increase people\u2019s awareness of their HIV status, non-symptomatic persons especially those in at risk populations should be screened frequently with tests that detect HIV infection before antibodies can be detected. Upon diagnosis, ART is to be initiated immediately , and virC-THAN will incorporate clinical and user needs in the development process for POC tests while providing access to expertise and resources to address early challenges to commercialization and implementation. The range of POC technology applications will include detection and monitoring of HIV/AIDS infection and its common fatal comorbidities including tuberculosis, infections of non-tuberculous mycobacteria, hepatitis B, hepatitis C and HIV-related malignancies.C-THAN\u2019s efforts entail: 1) assessment of unmet clinical needs in POC testing for HIV/AIDS and its comorbidities; 2) collaboration with relevant scientists, physicians, researchers and engineers; 3) development of essential technical, clinical, industrial and regulatory partnerships; 4) testing of prototype POC devices in the field; and 5) creation of training opportunities for technology developers, evaluators, and other stakeholders. Product development requires the coordinated activity of the four C-THAN cores: Administrative, Technology Training and Dissemination, Technology Development/Refinement, and Clinical Translation and Validation as illustrated in Administrative Core maintains the operational and strategic structure to manage C-THAN as a distinct cohesive entity that integrates, supports and coordinates partner and Core activities and supported projects. The Administrative Core leads the organization and governance of C-THAN, all budgetary matters, collaboration with POCTRN, communication with all stakeholders, and program/project monitoring and evaluation. Product development and commercialization requires the coordinated efforts of the Technology Development, Clinical Translation and Validation, and Technology Training and Dissemination Cores , University of Cape Town and Stellenbosch University (South Africa), University of Sciences, Techniques and Technologies Bamako and Muhimbili University of Health and Allied Sciences (Tanzania). C-THAN also has three biomedical engineering sites: Universities of Lagos, Ibadan and Cape Town. Investigators at these sites collaborate with Northwestern University faculty to provide the in C-THAN core functions described above and are available for individual collaborations. C-THAN provides opportunities for interactions with core members including potential collaborations in clinical needs assessment, conducting clinical validation studies, technology implementation, network building, and mentorship in project development, technology development, designing for future manufacturing, and grant writing.In each of the five years of the project, one-year technology development grants of approximately $100,000 will be awarded for HIV/AIDS related assay/device development projects. The grants must address the NIH\u2019s high priority topics for HIV/AIDS research, including: reducing HIV incidence by improving screening, detection, treatment monitoring, and HIV drug resistance detection; diagnosing HIV-associated comorbidities including tuberculosis, non-tuberculosis mycobacteria, hepatitis B, hepatitis C and HIV-associated cancers; reducing health disparities by developing testing technology for underserved community settings; and training of the workforce to translate POCT from research & development to implementation.in vitro diagnostic assays and technologies, treatment related diagnostic technologies, and technologies that improve or enable POC test performance. Essential product attributes include: meeting clinical needs of people living with HIV/AIDS; being operable in settings with limited medical infrastructure; and being simple to operate, durable, and manufacturable at low cost and with low-cost disposables.Six projects were selected for funding in 2019 (year two). Combined with the seven pilot projects in the original grant application, C-THAN\u2019s research portfolio currently covers https://cimit.org/ja/web/poctrn/home).Development grant awards for years three to five will go through a two-stage process. First, there will be a two-page pre-proposal (expression of interest) submitted with a description of the unmet clinical need, proposed innovation and collaborators, which will be triaged by C-THAN leadership with oversight of NIH scientific officers. The second phase will be submission of invited full proposals. The solicitation for proposals will be published on the POCTRN website (The content of the proposals will include a discussion of the unmet need and a description of the proposed solution including advantages over the alternatives and the maturity of the proposed solution. The work plan will include a table of deliverables and the implementation pathway. The application will also include a description of the team and resources, budget and justification. Funded projects that achieve their proposed milestones will be encouraged to apply for follow-on funding and/or clinical evaluation studies.Meeting UNAID\u2019s HIV 90-90-90 goals requires new approaches to successfully transform healthcare delivery to people living with HIV/AIDS. C-THAN\u2019s novel partnership model supports collaborations across disciplines, institutions and geographic regions and aims to drive innovative POC solutions. C-THAN seeks collaborators for technology development, clinical implementation, manufacturing and regulatory processes."} +{"text": "The reduction of environmental and health risks from pesticide use is on top of the agenda of the food-value chain actors. The establishment of pesticide-free production systems could be a cornerstone for the sustainable intensification of agriculture. In Switzerland, a pesticide-free but non-organic wheat production standard is currently being introduced. We present survey data of 1105 IP-SUISSE producers on adoption, future adoption, expectations and perceptions of the program, structural farm, and farmers' characteristics, and producers' risk preferences, farming objectives, attitudes and goals, self-efficacy, and locus of control. The data was collected to identify adoption determinants, barriers, and incentives for pesticide-free wheat production. The data is combined with publicly available data on soil properties, climate, weed pressure, and spread of herbicide resistance. We here provide data on the adoption of pesticide-free wheat production in Switzerland, which gives a comprehensive overview of potential adoption determinants and barriers for pesticide-free production.\u2022Researchers, policymakers and food-value chain actors aiming to understand the determinants and barriers of pesticide-free production can profit from this dataset.\u2022This data can be used in the future to provide further inisghts on socio-economic, environmental and farmers intrinsic drivers for the adoption of sustainable production practices in agriculture1We collected survey data from IP-SUISSE wheat producers in Switzerland. More specifically, we focused on the adoption of a pesticide-free but non-organic wheat production standard in the growing season 2019/20. The standard was first introduced as a pilot in the growing season 2018/19 and was opened up for public participation in the growing season 2019/20. The survey was designed based on a conceptual adoption model discussed with farmers, extension service experts, and farm advisors. The initial survey was pre-tested with ten producers. It was sent out to the entire population of IP-SUISSE wheat producers (4749). IP-SUISSE producers already produce wheat under an extensive production scheme of the producer organization IP-SUISSE, which prohibits the use of fungicides and insecticides in wheat production. The pesticide-free production scheme goes one step further and prohibits the use of all synthetic pesticide in wheat production. The survey data comprise 1105 complete responses (response rate of 23.3%) of producers. Survey data include information on adoption as well as potential adoption in the future.Moreover, expectations and perceptions of the program and farm and producers' characteristics were surveyed. We further collected producers' risk preferences, farming objectives, attitudes, goals, and non-cognitive skills such as self-efficacy and locus of control. Survey respondents were distributed all over Swiss wheat production regions see . We combhttps://doi.org/10.3929/ethz-b-000450297, including:-A readme file indicating all data files in the repository entry in text format.-An overview file, shortly describing variables of the survey data in PDF format.-The here described survey data with original variable names in csv format.-The here described survey data with English variable names in csv format.-The original questinnaires used (in German and French) in PDF format.-The original questionnaires translated in English in PDF format.All survey data and metadata are available from the ETH Z\u00fcrich Research Collection: We combined survey data with external data sources based on farm location. See 2The online survey link was distributed in December 2019 via E-Mail to all IP-SUISSE wheat producers in Switzerland (4749). At this point, producers had already decided to participate in the program for the growing season 2019/20. In the E-Mail, producers were encouraged to participate by IP-SUISSE. Furthermore, participation was incentivized with a lottery of 20 times 50 CHF supermarket vouchers among those participants completing the survey. The E-Mail highlighted that participation, both of producers, with positive and negative attitudes towards the program, is welcomed. We used Limesurvey to conduct the study and collect the survey data. Extension service experts and experts from IP-SUISSE reviewed the survey. It was then pre-tested with 10 IP-SUISSE wheat producers. The survey was online for five weeks, and two reminders were sent out after 2 and 4 weeks, respectively. In the survey, producers could choose between a German and a French version of the survey. In the introduction to the survey, its purpose was clearly explained again. Participants were reassured at the beginning of each of the three survey sections that their answers will be treated completely anonymous.We merged the survey data with publically available data on soil properties, climate, weed pressure, and spread of herbicide resistances from Agroscope by farm location , respectively.1.Participation and future participation in the IP-SUISSE program for pesticide-free wheat production and expectations towards the program's effects.2.Personal characteristics of the producer and characteristics of the farm3.Preferences and perceptionsThe survey contained 22 questions, and answering the questionnaire took producers a median time of 17.9 minutes. The structure of the survey was as follows:2.1In this part of the questionnaire, producers were first asked to state if they had participated in the pesticide-free wheat production program in 2018/19, if they were inscribed for 2019/20 or if they intend to participate in the future. The following questions were reframed depending on their answers, referring either to experiences or plans when adopting the program. Furthermore, farmers were asked about participation in direct payment programs for soil conservation and pesticide use reduction . We then asked them to evaluate decisive factors to participate/not participate in the program on a scale from 1-5, to state their experience with pesticide-free wheat production, their (intended) strategy for weed management under adoption, and availability of machinery to deploy these strategies. In the next questions of this part of the survey, we asked producers to state their perceived risks for machinery investment on a scale from 1-5, to evaluate the main factors for these risks, again on a scale from 1-5, and to indicate their main information sources for weed management. We concluded this section with questions about the producers' expectations concerning potential yield reduction and production risk increases under program adoption.2.2This section of the survey asked producers to indicate on-farm workforce, which part of their wheat production surface was leased, income shares in arable, animal, off-farm, and other production, education, age, and farm succession.2.3In the third section of the survey, producers were first asked to self-elicit their risk preferences on a scale from 0-10 in the domains of plant protection, agricultural production, marketing, and general decisions on the farm. Afterward, we asked them to evaluate their perception of the program's effects on the environment and human health, farming objectives, environmental preferences in farming, and openness for innovations and consultations with neighbors on a scale from 0-5. We then asked two questions, each concerning the producers' self-efficacy and locus of control, framed on wheat production and plant protection, respectively. Finally, we gave participants the possibility to leave comments.All methods were carried out in accordance with guidelines and regulations at ETH Zurich, and informed consent was obtained from all subjects prior to entering the survey. The project and study were approved by the office of research of ETH Zurich, and the survey was approved by the board of IP Suisse, the farmers\u2019 association representing participants of the study.Niklas M\u00f6hring: Conceptualization, Methodology, Validation, Data Collection, Data Curation, Writing \u2013 Original draft and Review &Editing, Visualization, Funding acquisition; Robert Finger: Conceptualization, Methodology, Data curation, Writing \u2013 Original draft, Funding acquisition.The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper."} +{"text": "In the original article, we neglected to include the funder, University of Zurich, UZH Postdoc Grant, grant no. [FK-19-040] to GF-G. The corrected Funding statement appears below.https://www.abmp.eu/). Additional funding was received from University of Zurich, UZH Postdoc Grant, grant no. [FK-19-040] to GF-G.This project was part of the research program \u201cCharacterization of functional brain network organization in dyslexia and development\u201d funded by the Amsterdam Brain and Mind Project, a UvA-VUA Amsterdam Academic Alliance Initiative (gorka.fragagonzalez@uzh.ch.In the original article the email address of the corresponding author was incorrect. The correct email address is The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Sci., 2020, 11, 6085\u20136096, DOI: 10.1039/D0SC02066D.Correction for \u2018Mechanistic details of the cobalt-mediated dehydrogenative dimerization of aminoquinoline-directed benzamides\u2019 by Li-Ping Xu The authors regret the omission of one of the authors, Manjaly J. Ajitha, from the original manuscript. The corrected list of authors and affiliations for this paper is as shown above.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers."} +{"text": "Correction to: BMC Health Serv Res 21, 1225 (2021)https://doi.org/10.1186/s12913-021-07229-yFollowing the publication of the original article , some inThe updated Funding and Disclaimer sections are given below.DisclaimerSome of the authors are present staff members of the World Health Organization. The authors alone are responsible for the views expressed in this article and they do not necessarily represent the views, decisions or policies of the institutions with which they are affiliated.FundingThis work was funded by the UNDP-UNFPA-UNICEF-WHO-World Bank Special Programme of Research, Development and Research Training in Human Reproduction (HRP), a co-sponsored programme executed by the World Health Organization (WHO).The original article has been"} +{"text": "Nature Communications 10.1038/s41467-021-26997-z, published online 18 November 2021.Correction to: The original version of this Article contained an error in Fig. 2, in which two curved arrows were inadvertently omitted. The correct version of Fig. 2 is: which replaces the previous incorrect version: This has been corrected in both the PDF and HTML versions of the Article.The original version of this Article contained errors in Equations (B), (C), (E), (G), (H) and (I). In Equation (B), the l of Cl was superscripted and a hyphen was not superscripted, and incorrectly read: In Equation (C), a dot was omitted, and incorrectly read: In Equation (E), the l of Cl was superscripted, and incorrectly read: In Equation (G), a space was added, and incorrectly read: In Equation (H), a 2 was not subscripted, and incorrectly read: In\u00a0Equation (I), a space was added and a 6 was not subscripted, and incorrectly read: This has been corrected in the PDF and HTML versions of the Article."} +{"text": "The Social Sciences section of The Journals of Gerontology, Series B: Psychological Sciences and Social Sciences aims to publish the highest quality social scientific research on aging and the life course in the U.S. and worldwide. The disciplinary scope is broad, encompassing scholarship from demography, economics, psychology, public health, and sociology. A key substantive focus is identifying the social, economic, and cultural contexts that shape aging experiences worldwide. In the coming decade, social gerontology research is poised to present many opportunities for cross-national and cross-cultural scholarship \u2013 driven in part by the proliferation of large parallel data sets from many nations in Europe, Latin America, and Asia. I will discuss the role that peer-reviewed cross-national scholarship can play in disseminating knowledge that informs gerontological research, policy, and practice internationally. I will also identify under-researched areas that will be of great interest to scholars in the coming decade, including LGBT older adults, aging in the Global South, reconfigured families, and centenarians."} +{"text": "Lifestyle includes routine and daily living activities affecting an individual's health. The present study aimed at evaluating the health-promoting lifestyle profile (HPLP) of medical sciences students of Kermanshah, Iran. In this cross-sectional study, 343 medical sciences students were enrolled by the stratified random sampling method. The data collection tools were demographic information form and the HPLP-II questionnaire. Data were analyzed using descriptive and analytical statistics. P \u2264 0.05). The mean overall HPLP-II score of the subjects was 2.25\u2009\u00b1\u20090.44 out of 4. Of the six HPLP-II dimensions, the highest and lowest scores belonged to interpersonal relations and physical activity, respectively. The mean overall HPLP-II score was statistically different in terms of gender, marital status, smoking habits, and economic status ( HPLP-II level was moderate in most of the students, and health-promoting behaviors, in the physical activity dimension, were in a low state. The results emphasized the need for interventions to improve students' lifestyles. Lifestyle includes daily routines that become habitual and affect physical and mental health . MeasureThis descriptive-analytic cross-sectional study was conducted in six faculties of Kermanshah University of Medical Sciences. The study was performed according to STROBE instructions.What are the mean scores of HPLP-II and its dimensions?What is the relationship between the mean HPLP-II and the personal characteristics of students?In this study, we sought to answer the following questions:n\u2009=\u20093725), including medicine, nursing and midwifery, health, paramedical, pharmacy, and dentistry. The sample size was calculated based on the results of the study of Rezaei-Adaryani and Rezaei-Adaryani [n\u2009=\u2009((\u03c3)2\u2217Z\u03b1/2)(2)/d2 equal to 312 people . To cover the missing data, 31 people (10%) were added to the sample size . The sample size was proportional to the total number of students in each faculty. The number of students in each faculty was as follows: medicine, 95 people; nursing and midwifery, 52 people; paramedical, 88 people; health, 57 people; dentistry, 18 people; pharmacy, 33 people.The study population included all the students studying in the affiliated faculties of Kermanshah University of Medical Sciences . The stratified random sampling method was used to select the subjects, and each faculty formed a stratum. The simple random sampling method using a table of random numbers was employed to select the subjects in each stratum.The demographic information form and HPLP-II questionnaire were used as data collection instruments. The demographic information form included nine items on age, gender, field of study, place of residence, marital status, economic status, weight, height, and smoking habits. The HPLP-II questionnaire was used to evaluate HPLP. Studies by Savarese et al. and Petrash & Murtazina evaluated the internal consistency of the questionnaire and reported its Cronbach's alpha as 0.94 and 0.88, respectively , 19. TheThe HPLP-II includes 52 items scored based on a four-point Likert scale as never , sometimAfter obtaining approval from the Ethics Committee of KUMS, the researcher referred to the Education Department of the affiliated faculties to enroll the eligible students. Accordingly, the study objectives were explained to students, and their consent for participation in the study was obtained. Then, the questionnaires were distributed among the subjects and collected after completion.Data were analyzed using SPSS software version 16. Descriptive statistics and analytical statistics were used to analyze the data. Kolmogorov\u2013Smirnov test was used to evaluate the normality of the HPLP-II variable and its subset, and the results showed an abnormal distribution of these variables.U test was utilized to compare the mean HPLP-II scores in terms of the two-state qualitative variables, including gender, marital status, place of residence, and smoking habits. The Kruskal\u2013Wallis H test was used to compare the mean HPLP-II scores in terms of the multistate qualitative variables, including the field of study, BMI, and family economic status. The level of significance for all tests was less than 0.05.The Mann\u2013Whitney The ethics committee of Kermanshah University of Medical Sciences approved the study with the code KUMS.REC.1399.167 . Written and informed consent was obtained from all participants. Emphasis was placed on the confidentiality of participants' information.In the current study, the response rate was 100%. According to the results, 173 subjects (50.4%) were females, 323 (94.2%) single, 306 (89.2%) dormitory residents, and 95 (27.7%) medical students, and 218 (63.6%) had a medium family income . The meaAccording to the results, the mean overall HPLP-II score was at a moderate level, in line with the results of studies by Mehri et al. and Montazeri et al. in Iran, Al-Qahtani et al. in Saudi Arabia, and Al\u2010Kandari in Kuwait , 22\u201324. The results showed that married subjects had a higher mean HPLP-II score than single ones. The finding was consistent with that of Lolokote et al. in China and inconsistent with that of Mehri et al. in Iran , 26. It In the present study, the mean overall HPLP-II score of the male subjects was significantly higher than that of females, consistent with the results of studies by Mehri et al. in Iran, Almutairi et al., Dhiman and Chawla, and Alzahrani et al. in Saudi Arabia and Paudel et al. in Nepal , 28, 29.The results showed that students who lived with their families had a higher mean overall HPLP-II score than the ones living in dormitories. This result was consistent with that of Mehri et al. in Iran and inconsistent with that of Alzahrani et al. in Saudi Arabia , 12. StuIn the current study, the mean overall HPLP-II score of nonsmoking students was significantly higher than that of smoking ones, consistent with the results of Alzahrani et al. in Saudi Arabia, Lolokote et al. in China, and Aynaci and Akdemir in Turkey , 26, 30.In the present study, students with a high economic level had a significantly higher mean HPLP-II score than the ones with middle to low levels. These results are in line with studies by Pakseresht et al. in Iran, Alzahrani et al. in Saudi Arabia, Lolokote et al. in China, and Dhiman and Chawla in Saudi Arabia , 26, 28.In the present study, the mean HPLP-II score was at a moderate level in all the age groups, and no significant difference was observed among them. This finding was consistent with the result of Alzahrani et al. in Saudi Arabia and incoThe results showed that the mean overall HPLP-II score had no significant differences among BMI groups , consistent with the results of Alzahrani et al. in Saudi Arabia and incoIn the present study, the mean HPLP-II score was at a moderate level in students of all KUMS faculties, and no significant difference was found among them. In the study by Pakseresht et al. in Iran, among medicine, dentistry, health, nursing, and paramedical faculties, the paramedical faculty had the highest mean score in interpersonal relations and spiritual growth dimensions . The stuIn the present study, among the six HPLP-II domains, the highest and lowest scores belonged to interpersonal relations and physical activity, respectively, consistent with the study by Mak et al. in China , althougThis study faced several limitations. One limitation was the possibility of social desirability bias. In this type of bias, instead of giving correct answers to questions, respondents give answers that are more acceptable to others and society. Considering the possibility of social desirability bias, the questionnaires were anonymous to minimize the impact. The main limitation of the cross-sectional study is that the temporal relationship between the exposure and outcome variables cannot be determined because the exposure and outcome variables are measured simultaneously. Therefore, due to the nature of cross-sectional studies, it is not possible to determine the causal relationship between the variables of this study. Considering the effect of personal and cultural variables on HPLP-II, caution should be exercised in generalizing the results.Most students had moderate HPLP-II. The highest and lowest scores of HPLP-II dimensions belonged to interpersonal relations and physical activity, respectively. There was a significant relationship between the mean HPLP-II score and the variables of gender, marital status, economic status, and smoking habits; however, it was insignificant in terms of BMI and age groups. It is suggested to evaluate the effect of interventions on students' HPLP-II in future studies."} +{"text": "Sarcopenia, an age-related degenerative disease, seriously affects the health and quality of life of the elder. The research of sarcopenia has changed dramatically around the world. This article aims to analyze global trends in this field over the past 20 years.\u201cSarcopenia\u201d was used as the search term to retrieve relevant publications from the WOS and PubMed databases. Co-occurrence, literature coupling, co-citation, and co-author analysis were performed by using the software VOS viewer. We analyzed the trends of sarcopenia research over the last 20 years from different aspects, such as the number of papers, total citations, average citations per item, h-index, research area, article types, institutions, country, journals, and funding.We retrieved 13,421 research articles published on sarcopenia between 2001 and 2020. The results showed that the USA made the highest contributions to this field. Geriatrics gerontology is the most study classification of sarcopenia. Basic research on sarcopenia in geriatric gerontology accounts for approximately 16.496% of global publications. The Osteoporosis International published the largest number of sarcopenia-related studies. The United States Department of Health Human Services was the leading funding organization, which sponsored 1,604 articles.Global sarcopenia research increased rapidly from 2001 to 2020, especially recently. The research leader of sarcopenia is the USA. In the future, the study of sarcopenia will continue to focus on aging, nutrition, and exercise and will delve deeper into molecular mechanisms. On the other hand, revealing the link between sarcopenia and other diseases will be the next research hotspot. Age-related decline in skeletal muscle mass was first defined as sarcopenia by Irwin Rosenberg in 1989, which is associated with an increased risk, such as physical disability, fracture caused by fall, and so on . In addiA study showed the prevalence of sarcopenia was 14.1% in Japanese residents aged 65 years or older . In addiBibliometric analysis of the sarcopenia-related research was conducted on August 21, 2021. Sarcopenia-related articles published from 2001 to 2020 were retrieved from the PubMed database and the Web of Science online database. Science Citation Index Expanded (SCI-EXPANDED), Social Sciences Citation Index (SSCI), Emerging Sources Citation Index (ESCI), Conference Proceedings Citation Index-Social Science and Humanities (CPCI-SSH), Conference Proceedings Citation Index-Science (CPCI-S), and Arts and Humanities Citation Index (A&HCI) are all included. The impact factor (IF) of journals came from the journal citation reporting database. The data in this study are derived from a public database and no ethical issues are involved. Therefore, this study was not reviewed by the ethics committee.In the Web of Science online database, the search terms were: topic = sarcopenia and publishing year = (2001\u20132020). The search results were refined by articles, review, meeting abstract, letter, and proceedings paper to identify the core literature for this bibliometric analysis. In the PubMed database, the search term \u201csarcopenia (Mesh Terms)\u201d does not retrieve all the publications concerning sarcopenia. Consequently, \u201csarcopenia\u201d (All Fields) and results from 2001 to 2020 can retrieve all publications concerning sarcopenia. The document types retrieved included clinical trial, meta-analysis, randomized controlled trial (RCT), review, systematic review, case reports, and basic research studies. To find basic research studies, the species was identified as an \u201cother animal\u201d. Using the total reference frequency, the average number of references per item, and the h-index to assess the publication quality. The quantity of literature and publication trends were analyzed based on the total amount of publication, research organization, research field, type of articles, and journals.Information of all the eligible publications, including the title, author's name and affiliation, nationality, keywords, year of publication, and the journal, was selected from the scientific online database. All the authors screened, collected, and extracted the data, resolved the differences, and reached a consensus.P < 0.05). All these were conducted by SPSS software .The descriptive statistical analysis in this study was analyzed by using linear regression from 39 in 2001 to 2,767 in 2020. Most research was published in 2020 .Based on search criteria, there are 13,421 articles, including article , review , meeting abstract , letter (416), and others, were published in the world from 2001 to 2020 in the Web of Science. As There are 105 countries/regions that participated in sarcopenia-related research around the world. Among them, the largest number of sarcopenia articles are published by the USA , followed by Japan , Italy , England , and South Korea . The annAccording to the data from the Web of Science, sarcopenia papers from the USA possessed the highest total citation frequencies , followed by Italy , England , and Canada . For the average citation frequencies per item, Sweden ranked first (110.48), followed by Switzerland (81.09), Scotland (73.44), Netherlands (61.85), and Belgium (61.43). In terms of the H-index, the index of the USA was 180, which was ranked first, followed by Italy with an index of 105 .A total of 7,287 institutions from different countries/regions contributed to sarcopenia-related research between 2001 and 2020. Globally, sarcopenia-related publications were categorized into 74 research areas. According to the PubMed database, basic research was popular among researchers. There are 1,835 basic research articles in the past 20 years, accounting for 16.496% of the total sarcopenia articles. Additionally, 716 clinical trials (6.437%), 584 RCTs (5.250%), and 53 case reports (0.476%) were published in the sarcopenia field .Co-occurrence analysis can be used to discover directions and popular topics of specific areas, which is vital for monitoring the development of science and programs. From the global keyword's maps, the co-occurrence networks of keywords for 2001, 2004, 2008, 2012, 2016, and 2020 are shown in As can be seen from Bibliographic coupling analysis is a method that uses citation analysis to establish a similarity relationship between different documents. The name of journals in total publications was analyzed by VOS viewer. In total, 203 institutions (Minimum number of documents of an organization: 30) were analyzed in this study . The UniThere are 74 countries (Minimum number of documents of a country: five) that were analyzed in this study . The USACorrelations between items can be identified by the co-citation analysis by the number of times they were cited together. In total, 924 references (Mini number of citations: 50) are analyzed in our study . Cruz-jeIn total, 409 journals (mini number of citations of a source: 200) are analyzed as shown in Co-authorship analysis judged the relevance of the items by the number of its co-authored papers. In total, 245 authors (Minimum number of documents of an author: 15) were analyzed in our study . The topIn Publications (defined as the minimum number of documents of a country that were used more than five) identified in the 74 countries were analyzed by VOS viewer . The topA total of 2,019 journals published sarcopenia articles. The number of articles published has increased significantly over the years, which has made keywords more prevalent. Studies through Bibliometric and visualized research can be used to show the status of research in a certain field and to predict future trends. As the global population ages, the prevalence of sarcopenia will increase. Therefore, there will be more institutions dedicated to sarcopenia research. Additionally, more studies on sarcopenia will be published in the next few years.By the co-occurrence analysis, we found that directions and research focus in this field. The keywords as the most important part of articles were used to create a co-occurrence network map. From The total citations and h-index of countries represent their academic influence and publication quality. The USA contributed the most to global sarcopenia-related research with high total publications and total citations, especially the total citations of the USA were far was ahead of other countries. As a result, the US can be considered a leader in this field. Osteoporosis International published the largest number of publications, which is the most contribution journal in the world. Of 10 journals publishing the largest number of reports, three journals are from the UK, two journals are from the USA, and the rest are from Germany, France, Switzerland, Scotland, and Australia. The journals on the list may be included the main discoveries in this field in the future. Institutes from the top five countries were the leader in sarcopenia-related research. All of the top 15 institutes were located in the top nine countries. The United States Department of Health Human Services and the National Institutes of Health were the main funding organization, whose contribution was far more than any others. The top 10 authors who published most articles in sarcopenia were also listed and we can keep track of the scientific frontiers of this field by paying close attention to their published work. At present, the research on sarcopenia is mainly focused on geriatrics gerontology and basic research. In this study, a similar relationship between publications in terms of journal, institution, and country was established through bibliographic coupling analysis. Bibliographic coupling occurs when two works cite a common third work in their articles . The datThis study had some limitations. There are some limitations on the methodological quality of our study, Future improvements in methodology are necessary. First, bibliometric analysis can only measure the quality of scientific research through the influence index of literature. However, there is no absolute equivalence between them. For example, a highly cited publication does not mean high scientific quality. Second, the data in our study were from the PubMed database and the Web of Science online database. Therefore, we may omit some other literature. Third, we only selected literature written in English as the research objects, excluding many non-English publications. Finally, some recent research of high scientific quality might not be emphasized due to fewer citations. Therefore, paying attention to the latest publications is also necessary.As the research field of sarcopenia becomes a hotspot, the future research trend of sarcopenia will receive researchers' extensive attention. Sarcopenia, obesity, and sarcopenic obesity are vital characteristics of the aging process that can lead to important health problems. Future studies may focus on the interrelationships between these changes in body composition . MalnutrGlobal sarcopenia research increased rapidly from 2001 to 2020, especially in recent years. The USA was the leader of sarcopenia-related research. The study of sarcopenia will continue to focus on aging, nutrition, exercise, and delve deeper into molecular mechanisms in the future. In addition, revealing the link between sarcopenia and other diseases may be the next research hotspot.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.DY and HJ participated in study design, data collection, statistical analysis, and manuscript preparation. QL, JZ, and BM participated in the data check. WX and YL conceived and revised this article. All authors approved the final version of the manuscript.This study was supported by the National Natural Science Foundation of China (Nos. 81874030 and 82072506), Science and Technology Innovation Program of Hunan Province (No. 2021RC3025), Provincial Natural Science Foundation of Hunan (No. 2020JJ3060), Provincial Clinical Medical Technology Innovation Project of Hunan (No. 2020SK53709), Innovation-Driven Project of Central South University (No. 2020CX045), Wu Jieping Medical Foundation (No. 320.6750.2020-03-14), National Clinical Research Center for Geriatric Disorders , National Clinical Research Center for Orthopedics, Sports Medicine and Rehabilitation (No. 2021-NCRC-CXJJ-PY-40), Exploration and Innovation Project for Undergraduate Students of Hunan (Nos. S2021105331047 and S2021105331107), and Central South University (No. XCX2021046).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Anneissia intermedia is a common crinoid found in waters along the coastlines of China, Japan, and Korea. In this study, we determined the complete mitogenome of A. intermedia. The genome was found to be 15,874\u2009bp in length and consists of 13 protein-coding genes, 22 tRNAs, and 2 rRNAs. With the exception of Antedon mediterranea, the gene order and genetic characteristics of the A. intermedia mitogenome are identical with those of the mitogenomes of other crinoids. The complete mitogenome of A. intermedia will contribute to enhance our understanding of the phylogeny of crinoids. Anneissia consists of nine species distributed over a wide range of the Indo-western Pacific Ocean, including Australia, New Zealand, New Caledonia, China, Japan, and Korea . Voucher specimens and mitochondrial DNA (mtDNA) samples have been deposited at the Marine Echinoderm Resources Bank of Korea under the voucher number MERBK-C-00307. mtDNA extraction, amplification, and sequencing were performed in accordance with the methods described by Lee and Shin (A. intermedia), four asteroids, four ophiuroids, four echinoids, and three holothuroids], with those of two hemichordates being used as an outgroup. Mitogenome sequences were aligned using MAFFT (Katoh and Standley The genus and Shin . The comand Shin . PhylogeA. intermedia (GenBank accession No. MW376476) is 15,874\u2009bp in length and contains 13 PCGs, 22 tRNA genes, and two rRNA genes. With the exception of that of Antedon mediterranea, the gene order and direction of PCGs and rRNAs were found to be identical to those of the complete mitogenomes of other comatulid species. All PCGs contain an ATG initiation codon (methionine), and the termination codons are generally either TAA or TAG, with that of cytochrome b, which terminates with (GAG)+T (glutamic acid), being the only exception.The complete mitogenome of A. intermedia clusters with A. pinguis and Phanogenia gracilis, belonging to the same family, Comatulidae (A. intermedia will provide a basis for further phylogenetic studies on crinoids.In the constructed phylogenetic tree, the nine complete mitogenomes of crinoids clearly established a monophyletic clade , within atulidae . The seq"} +{"text": "Light: Science & Applications11, 6 (2022)10.1038/s41377-021-00694-4Two-color infrared detection technology realizes target recognition in a complex environment by using the multispectral characteristics of the target. In the last decade, several papers have announced the usefulness of the 2D materials for high operating temperature photodetectors covering infrared spectral regions. Researchers from Shanghai Institute of Technical Physics of Chinese Academy of Sciences, Huazhong University of Science and Technology, and Fudan University demonstrated an uncooled two-color infrared photodetector based on van der Waals heterojunction. This two-color photodetector can detect near-infrared and mid-wave infrared at the same time, and with ultra-low crosstalk, it realizes spectral blackbody detection with temporal and spatial coherence. Its room temperature operating ability greatly reduces the volume, weight, and power consumption of the detection components, and demonstrates the application prospects of van der Waals heterostructures in the miniaturized and intelligent photodetection systems."} +{"text": "The editorial board and the Brazilian Urology Society are very proud with consolidation of the International Brazilian Journal of Urology as one of the most relevant in the dissemination of urology research worldwide.This is a historical number for our Journal. We are pleased to announce that International Brazilian Journal of Urology reached the biggest impact factor of its history - Int Braz J Urol, the 13th under my supervision, presents original contributions with a lot of interesting papers in different fields: Renal Cell Carcinoma, Bladder Cancer, SARS-CoV-2 and Urology, Basic Research applied to prostatic diseases, Premature ejaculation, Reconstructive urology, Lower urinary stones, Ureteral Stones, Lower urinary tract symptoms in children and Xanthogranulomatous Pyelonephritis. The papers came from many different countries such as Brazil, USA, Iran, Israel, Colombia and Singapure, and as usual the editor's comment highlights some of them.The September-October 2021 number of Dr. Sharma and colleagues from India performed in page 921 (Dr. Dispagna and colleagues from USA present Dr. Zekan and colleagues from Department of Genitourinary Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL, USA under supervision of Dr. Philippe Spiess present Dr. Alam and colleagues from, USA present Dr. Vasconcelos and colleagues from Brazil present Dr. Falahatkar and colleagues from Iran present Dr. Frumer and colleagues from Israel present Dr. Anaissie and colleagues from, USA present Dr. Oliveira and colleagues from Brazil present Dr. Favorito and colleagues from Brazil present We hope that readers will enjoy the present number of the International Brazilian Journal of Urology in this very difficult times of COVID-19."} +{"text": "There has been an important global interest in Open Science, which include open data and methods, in addition to open access publications. It has been proposed that public availability of raw data increases the value and the possibility of confirmation of scientific findings, in addition to the potential of reducing research waste. Availability of raw data in open repositories facilitates the adequate development of meta-analysis and the cumulative evaluation of evidence for specific topics. In this commentary, we discuss key elements about data sharing in open repositories and we invite researchers around the world to deposit their data in them. There is an important global interest in Open Science, which include open data and methods, in addition to open access (OA) publications , 2. Sevehttps://www.cos.io/initiatives/top-guidelines) [It has been proposed that public availability of raw data increases their value and the possibility of confirming scientific findings, improving reproducibility and replicability of results \u20138, in addelines) , 8. It hdelines) . Availabdelines) , and thedelines) , especiadelines) (such asdelines) .A recent study showed, in a sample of 531.889 OA journal articles, that a minor fraction of papers included a link to data repositories and that those articles have a higher citation impact . An analThe FAIR Guiding Principles have been proposed for scientific data management and theyhttps://creativecommons.org/about/cclicenses/) are frequently used [https://creativecommons.org/share-your-work/public-domain/cc0) should be used for data sharing.There are several available OA licenses and the ones from Creative Commons facilitates consolidation and search of open datasets from that region of the world [https://www.rd-alliance.org) [In Table he world . The Resnce.org) .Table 1Ihttps://www.nature.com/sdata).There is a need for more training about open science and data science , particuIn times of COVID-19, it is critical to have good quality data for proHowever, several challenges have to be solved, particularly in emerging economies, such as: legal and policy issues, scarcity of coordination between research groups, lack of a culture for data sharing and ethical/privacy considerations, insufficiency of proper infrastructure (including high-speed Internet connectivity), deficiency in interoperability of platforms, shortage of data managers and data scientists and a scarcity of open data repositories to facilitate data sharing . Recentl"} +{"text": "Journal of Patient-Reported Outcomes (JPRO) is an international, open access, multi-disciplinary journal publishing original research and review articles, brief communications, commentaries, editorials, and reviews of recent books and software advances in the field of patient-reported outcomes. The JPRO is an official journal of the International Society for Quality of Life Research (ISOQOL). The first JPRO articles were published on September 12, 2017.The We want to acknowledge the immense contributions of Dennis Revicki, JPRO Founding Co-Editor-in-Chief, who died on May 9, 2021. His impact on ISOQOL and the field of patient-reported outcomes was immense. We miss him greatly.University of Sheffield, UK), Joanne Greenhalgh , John Devin Peipert , Caroline Potter , and Jessica Roydhouse .The fifth year (2021) of the JPRO was another successful one due to the contributions made by reviewers and the consistently high quality of the science reflected in the published articles. We are also extremely grateful for the conscientious work of the associate editors: Jill Carlton (We are confident in a very positive trajectory for the JPRO in 2022 and we encourage those conducting patient-reported outcomes research to submit manuscripts and to serve as peer reviewers.Sincerely,Ron D. Hays and Chih-Hung ChangJPRO Co-Editors-in-Chief"} +{"text": "Dr. Praveen Kerala Varmaemail: varmapk@gmail.com; praveenv21204@aims.amirta.eduDr. Rajesh Joseemail: rajeshj21304@aims.amirta.eduFollowing publication of the original article , the autThe author group has been updated above and the original article has beenAuthors' contributionsJJ: developed the graft in Amrita, did lab-based work in Bristol, analyzed data and drafted the manuscript; VDB: assisted animal surgery in Bristol and undertook vascular doppler; NS: helped with lab-based work in Bristol and the bioreactor; AW: helped with graft engraftment lab-based work in Bristol; TJ: undertook the ex-vivo optical coherence tomography on explanted grafts in Bristol; HMB: undertook mechanical strength work at Amrita and prepared Fig.\u00a02a, 4a, and graphical abstract; PKV: helped developing the graft in Amrita before studies in Bristol; RJ: helped developing the graft in Amrita before studies in Bristol; SK: helped developing the graft in Amrita and securing previous funding from the Indian Government; DM: helped with developing the graft in Amrita including supervising the Amrita-based bench testing and related interpretation, co-secured the funding for the Commonwealth Scholarship Commission for the Split-site scholarship, co-drafted the manuscript; SJG: supervised the lab-based histology work in Bristol related to graft characterization and interpretation, co-secured the funding for the Commonwealth Scholarship Commission for the Split-site scholarship, co-drafted the manuscript; RA: acted as senior author, conceived and led all the bioreactor and in-vivo work done in Bristol and related interpretation, secured all funding to support bioreactor and in-vivo work in Bristol, co-drafted the manuscript and was corresponding author. All the authors have read and approved the final manuscript."} +{"text": "ABSTRACT IMPACT: The mobilization of a CTSA-sponsored team with multi-disciplinary translational science expertise enabled the university to provide a range of T1-T4 expertise to a large, complex school district that resulted in permanent learning and data science infrastructure. OBJECTIVES/GOALS: The Clinical Translational Science Institute (CTSI) formed a multidisciplinary science team to provide expertise in support of the re-opening of in-person learning in the second-largest U.S. school district during the COVID-19 pandemic. METHODS/STUDY POPULATION: The assembled interdisciplinary science team provided expertise in epidemiology, machine learning, causal inference and agent-based modeling, data and improvement science, biostatistics, clinical and laboratory medicine, health education, community engagement, and experience in outbreak investigation and management. The team included TL1 pre and postdoctoral fellows and mobilized scientists from multiple professional schools and T1-T4 stages of translational research. RESULTS/ANTICIPATED RESULTS: Tangible outcomes achieved using this team approach included the development of practical metrics for use in the school community, a learning process, the integration of preventive design elements into a testing and tracing program, and targeted and data-driven health education. The team, for example, generated new data displays for community engagement and collaborated with the school district in their use to visualize, learn from, and act on variation across a 700 square mile region. DISCUSSION/SIGNIFICANCE OF FINDINGS: Novel translational methods can be used to establish a learning environment and data science infrastructure that complements efforts of public health agencies to aid schools in the COVID-19 pandemic. These new capabilities apply to COVID-19 testing and vaccines and can be mobilized for future population health challenges faced by school districts."} +{"text": "Okada et al.). The aforementioned reviews suggest that changes in the directionality of water transport involve a variety of mechanisms; they can be osmotically obligated in their nature and coupled to the movement of ions and organic osmolytes, driven by hydrostatic pressure, or determined by \u201cingestion\u201d and \u201cexcretion\u201d during endo/exocytotic processes.The flow of water across cellular membranes determines the dynamics of cellular hydration and volume, both of which govern a plentitude of fundamental functions, including cell death (CD) , represents a hallmark of the unregulated form of CD\u2014necrosis . In contrast, the highly regulated mode of CD, apoptosis, is generally associated with cell volume decrease . Apart from the two major types of CD, apoptosis and necrosis, numerous additional (sub)forms of CD have been identified. These include aponecrosis, oncosis, necroptosis, parthanatos, anoikis, entotic CD, NETotic CD, immunogenic CD, lysosome-dependent CD, ferroptosis, oxeiptosis, sarmoptosis, autosis, autolysis, paraptosis, pyroptosis, alkaliptosis, phagoptosis, eryptosis, chondroptosis, autophagic CD, mitoptosis, methuosis, and the mitotic catastrophe-driven CD . Understanding these mechanisms is crucial for our comprehension of the basic principles of normal and abnormal biological processes.The significance of water transport and cell volume in CD has been recognized for a long time. Injurious cell swelling, which was initially described as oncosis , two Brief Research Reports , 11 Reviews , four Mini Reviews , one Hypotheses article , and one Opinion article These publications are contributed by 98 authors. While in the production, the Research Topic was met with high interest within the scientific community; it accumulates the steadily increasing number of views and downloads from all parts of the world (https://www.frontiersin.org/research-topics/13260/ion-and-water-transport-in-cell-death#impact).The objective of this Research Topic was to collect state-of-the-art Reviews and cutting-edge original research articles exploring ions and water transport in cell death. This timely subject has attracted significant enthusiasm of the scientific community. The initial Call for Contributions resulted in 23 accepted manuscripts covering various aspects of the pivotal roles of ion and water transport in cell death. The collection encompasses four Original Research papers , necrosis , aponecrosis , necroptosis and pyroptosis , ferroptosis , paraptosis , eryptosis , methuosis as well as plant vacuolar CD . From the standpoint of the mechanisms of CD-inducing processes, the Topic contributors discuss the functional significance of numerous specific anion channels, cation channels, and ion transporters . To facilitate reading of this collection, we provide cross-references to the related ion transport mechanisms in Bortner and Cidlowski; Shiozaki et al.), and mechanistic contributions for some organic signaling molecules which are released via anion channels .The articles in this Research Topic cover a large variety of types of CD, including apoptosis , chronic neurodegenerative disorders, including Alzheimer's disease , cancer , and chemoresistance of cancer ; necrosis in I/R injury including lactacidotoxicity , osteoarthritis , and cancer (Lefranc); aponecrosis in I/R injury ; necroptosis and pyroptosis in I/R injury, neurodegeneration, cancer, skin inflammation, and crystallopathies ; ferroptosis in I/R injury , neurodegeneration , cancer , and acute CNS injury ; paraptosis in I/R injury and chemoresistance of cancer ; as well as eryptosis in anemia and chronic kidney disease (CKD) . There is a hope in the field that further elucidation of molecular mechanisms of ion and water transport underlining these CD processes is likely to provide accurate targets for therapy of CD-associated diseases and for the treatment of chemo-resistant cancer.It is also important to place each CD type in the context of the pathogenesis of different human diseases. The contributors discuss the role of apoptosis in ischemia/reperfusion (I/R) injury, including excitotoxicity , the Japan Society for the Promotion of Science (Grant-in-Aid for Scientific Research #17K19517 to YO), POR Puglia Innonetwork , and Agenzia Spaziale Italiana (GV).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Background: Though Gilles de la Tourette's syndrome (GTS) has significant impact on the quality of life of its patients, measures of health-related quality of life (HR-QOL) specific to adolescents and adults with GTS were not developed until recently. The present study provides evidence on the validity of the Gilles de la Tourette Syndrome-Quality of Life Scale (GTS-QOL), the first disease-specific HR-QOL instrument for GTS patients, for the first time in an East Asian sample.Methods: One hundred and two Japanese individuals aged 13 and above with GTS were included in our study. Internal consistency was evaluated using Cronbach's alpha. The 4-factor structure of the GTS-QOL was assessed using confirmatory factor analysis, using goodness of fit indices, factor loadings of each questionnaire item, and covariances between factors. Validity was assessed using interscale correlations. Convergent and discriminate construct validity was evaluated using correlations with other scales such as the 28-item General Health Questionnaire, the Yale Global Tic Severity Scale, and the short version of the Padua Inventory.Results: Scaling assumptions were met. Internal consistency reliability was high, with a Cronbach's alpha of 0.96. Confirmatory factor analysis revealed sufficient factor loadings and goodness of fit. All measures of goodness of fit corroborated the fit of the 4-factor model. Standardized covariances between factors in the confirmatory factor analysis were >0.8. There were significant correlations with other well-validated scales, and thus convergent and discriminate construct validity was sufficient.Conclusion: The GTS-QOL is a valid and reliable instrument to measure disease-specific HR-QOL of GTS patients in Japan. Gilles de la Tourette's Syndrome (GTS) is a neurological disorder characterized by multiple motor and vocal tics that often appears in childhood and affects up to 1% of school age children , and GTSDespite the growing evidence of lower HR-QOL in patients with the disorder, a HR-QOL measure specific for GTS that addresses the various facets of the disorder that have significant impact on the patient's QOL was not introduced until recently . The firThe GTS-QOL overcame the major shortcoming of applying generic HR-QOL instruments to HR-QOL of GTS patients; these instruments fail to address features specific to GTS patients that may cause distress, such as motor and vocal tics and repetitive behaviors . A previParticularly, though generic measures of HR-QOL have the advantage of allowing comparison across different diseases, they fail to address features unique to GTS, such as repetitive behaviors, motor and vocal tics, and coprolalia, as well as common comorbidities and problems in social life. On the other hand, assessment of disease status by clinical rating scales of tic severity alone omits the patient's subjective views about issues important to their lives, especially cognitive and socioemotional functioning. The development of the GTS-QOL was significant in that it conquered these shortcomings and provided clinicians and researchers with a tool to better characterize each GTS patient and monitor the clinical course or treatment effects over time.Since its introduction in 2008, multiple versions of the GTS-QOL have been validated , 15 and We included patients with GTS who were aged 13 years or older who fully answered the GTS-QOL (including the GTS-QOL VAS).The Japanese version of the GTS-QOL, including the GTS-QOL VAS, was translated and back-translated by an experienced GTS researcher.The questionnaires were distributed to GTS patients who were members of the Tourette Syndrome Association of Japan and patients who attended the outpatient GTS clinic of the University of Tokyo Hospital or local psychiatry facilities in Tokyo and Kanagawa prefectures at the time of the study. The patients were asked to mail in or hand in the completed questionnaires. Additionally, outpatients of the University of Tokyo Hospital were asked to complete an interview by a psychologist for the assessment of the Yale Global Tic Severity Scale (YGTSS). Three hundred thirty-two patients were recruited from October 2019 to March 2020, and of these patients, we obtained informed consent from 95 of 112 (84.8%) patients from the University of Tokyo Hospital, 51 of 180 (28.3%) patients from TSAJ, and 28 of 40 (70.0%) patients from local psychiatry facilities in Tokyo or Kanagawa, making a total of 174 patients. Informed consent was obtained either by written consent after a participant received an explanation about the study or by answering \u201cyes\u201d to one of the questions in the questionnaire asking if the participant agreed to participate in the study .In addition to the GTS-QOL, we included questions from the following scales in the questionnaire. We used 28-item General Health Questionnaire (GHQ-28) , a self-We calculated the skewness and the ceiling and floor effects of the total GTS-QOL as well as the 4 subscales. We also calculated item mean scores, standard deviations, and item-total correlations.We calculated Cronbach's alpha to test the internal consistency reliability of the GTS-QOL.p-value, comparative fit index (CFI), root mean square error of approximation (RMSEA), and standardized root mean square residual (SRMR). Additionally, we evaluated standardized covariances between factors as well as interscale correlations of the subscales.We conducted a confirmatory factor analysis using the 4-factor model identified previously , in ordeP = 0.001, after Bonferroni correction for multiple hypothesis tests . The correlation between GTS-QOL and each scale was calculated using data from participants whose answers for each scale were fully available, and significance level was set at is tests .The study was approved by the ethics committee of the University of Tokyo [IRB number: 10183-(4)]. All statistical analyses were conducted using R version 3.6.3.Our study sample was comprised of participants 13 years or older, whereas the participants in the original GTS-QOL study were 16 years or older . In orde102 patients met the inclusion criteria. 75 participants (73.5%) were males, and 27 (26.5%) were females. The average age of the participants was 24.7 years . YGTSS and GAF were performed on a total of 40 patients. 29.4% of participants reported having comorbid OCD, and 21.6% reported having comorbid ADHD. The prevalence of these comorbidities were similar to what was reported in similar settings .All item response options were endorsed. Skewness of the total GTS-QOL was 0.35, and the skewness of the 4 subscales ranged from 0.40 to 0.72 . The ceiInternal consistency reliability was high, with a Cronbach's alpha of 0.96 .P < 0.001; GHQ-28 0.72, P < 0.001; YGTSS total tic severity 0.55, P < 0.001; GAF \u22120.55, P < 0.001; PI 0.77, P < 0.001; P = 0.001) was larger than between YGTSS motor tic severity . As for the subscales of GTS-QOL, the correlation between the psychological subscale and the GTS-QOL VAS as well as GHQ-28 was the strongest of the four subscales. YGTSS total tic severity was most strongly correlated with the physical/ADL subscale of the GTS-QOL . The PI total score was most strongly correlated with the psychological subscale , followed by the obsessive-compulsive subscale . Age and disease duration were not significantly correlated with GTS-QOL . Other results of the correlations with scales are shown in Convergent and discriminate construct validity was sufficient, with significant correlation between scales and satisfaction with life (GTS-QOL VAS) were most strongly correlated with the psychological subscale , 14, 15.A study on Japanese GTS patients used both quantitative and qualitative methods to show that HR-QOL was lower than the general population and that tic severity and comorbid ADHD were associated with lower HR-QOL . This fiCompared to the British study sample in the original GTS-QOL study by Cavanna et al., the mean GTS-QOL total score was greater in the present study's Japanese sample , despite the fact that the mean YGTSS total tic severity was smaller in the present study (20.4 compared to 26.8). The Japanese sample may have been more prone to feel more distressed about their symptoms compared to the British sample, even if the severity of their tic symptoms were similar. This could be explained by the strong psychological component of HR-QOL of Japanese GTS patients described previously, though further research is necessary to confirm this. Importantly, GTS-QOL may not necessarily have cross-cultural equivalence, and cultural context should be taken into account when evaluating the total GTS-QOL score.Our study is of great significance in that studies on the HR-QOL, let alone disease-specific QOL, of GTS patients is very limited. Furthermore, other than the aforementioned mixed method study on the HR-QOL of Japanese GTS patients, studies on HR-QOL in GTS patients are rarely conducted in Japan, and the present study should open the door to more studies in this field.Our study should be interpreted in light of several limitations. The patients were recruited at a tertiary referral hospital as well as a patient association, and this may have caused our sample from deviate away from the actual population of GTS patients in Japan. Furthermore, we were unable to perform test-retest reliability analysis since the questionnaire was administered only once to each patient, though our findings to support the reliability and validity of the Japanese version of the GTS-QOL, to the extent possible.Additionally, our study sample consisted of participants 13 years or older, whereas the participants in the original GTS-QOL study were 16 years or older . AlthougNotwithstanding the limitations, our study has multiple clinical implications and provides opportunities for future research. HR-QOL is important in measuring the impact of a chronic disease on the patient . This isAn effective evaluative instrument can detect important changes, large and small, over a period of time . In thisAs the discriminative and evaluative performance of the GTS-QOL is confirmed in additional studies, it can be used to collectively capture features of GTS other than tics, such as obsessive-compulsive symptoms, ADHD traits, depressive symptoms, and social functioning, over time. A study has shown that by early adulthood, tic symptoms will be greatly diminished in three-quarters of children with GTS and one-third will be tic-free . Hence, To date, the GTS-QOL (its child and adolescent variant) has only been applied to GTS patients as young as 7 years old . The onsThe validity and reliability of the 27-item GTS-QOL was confirmed in Japanese GTS patients.kano-tky@umin.ac.jp.The datasets presented in this article are not readily available because the Ethics Review Board has not authorized data sharing of this research. Inquiries regarding the data should be directed to Yukiko Kano, The studies involving human participants were reviewed and approved by the Ethics Committee of the University of Tokyo. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.RG, YY, and YK conceived the research idea. RG conducted the analyses and wrote the manuscript. YK directed the research. NM, MN, YH, YE, MF, AS, YY, and YK gave critical feedback on the research and approved the final manuscript. All authors contributed to the article and approved the submitted version.This work was supported by Health and Labor Sciences Research Grants for Comprehensive Research on Disability Health and Welfare from the Ministry of Health, Labor, and Welfare of Japan (Grant No. 19GC1001) and Grant-in-Aid for Scientific Research (C) (Grant No. 20K03435) from the Ministry of Education, Culture, Sports, Science and Technology in Japan.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "IKBKG variant in a 7-month-old boy with pneumococcal rib osteomyelitis and later found that his mother has incontinentia pigmenti. Genetic analysis of preserved umbilical cords revealed the same variant in two of his deceased maternal uncles. Analysis of preserved umbilical cord tissue from deceased relatives can provide important information for diagnosing IEI in their descendants.Family history is one key in diagnosing inborn errors of immunity (IEI); however, disease status is difficult to determine in deceased relatives. X-linked anhidrotic ectodermal dysplasia with immunodeficiency is one of the hyper IgM syndromes that is caused by a hypomorphic variant in the nuclear factor kappa beta essential modulator. We identified a novel Family history is one of the most important items of the 10 Warning Signs of Primary Immunodeficiency Diseases for the prediction of primary immunodeficiency diseases . HoweverIKBKG (on Xq28), which encodes the nuclear factor kappa beta (NF-\u03baB) essential modulator (NEMO). An IKBKG variant is also associated with incontinentia pigmenti (IP) in females. Although the typical variant in IP is lethal in males anhidrotic ectodermal dysplasia with immunodeficiency (EDA-ID) is a rare IEI. XL-EDA-ID is based on a hypomorphic variant of in males , hypomorKG on Xq2, which ein males . Herein,Haemophilus influenzae type b vaccine, hepatitis B vaccine, and pentavalent rotavirus vaccine. The patient was born to non-consanguineous parents, and he had been thriving without growth failure. Family history was significant for two maternal uncles who had died at ages 4 and 7 months Figure\u00a03IKBKG gene were used instead of long-range PCR , and minor(s)\u2019 legal guardian/next of kin, for the publication of any potentially identifiable images or data included in this article.Patient's management: SI, YA, CI, and AS. Manuscript preparation: SI and YA. Study concept: HK. Study Design: YA, HO, and HK. Literature search: SI, YA, and HK. Data analysis/interpretation: YA, YM, CI, HO, and HK. Manuscript editing: CI, HO, HK, and AS. All authors contributed to the article and approved the submitted version.This work was supported by Niigata Univ. Early-Career Scientists Development Program to YA and by MEXT KAKENHI Grant Number JP21K07770 and Health and Labour Science Research Grants for Research on Intractable Diseases from the Ministry of Health, Labour and Welfare of Japan (Grant Numbers 20316700 and 20317089), and by AMED (Grant Number JP20ek0109480) to HO.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "COVID-19 disease caused by SARS-CoV-2 represents an ongoing global public health emergency. Rapid identification of emergence, evolution, and spread of SARS-CoV-2 variants of concern (VOC) would enable timely and tailored responses by public health decision-making bodies. Yet, global disparities in current SARS-CoV-2 genomic surveillance activities reveal serious geographical gaps. Here, we discuss the experiences and lessons learned from the SARS-CoV-2 monitoring and surveillance program at the Public Health Laboratory on Bioko Island, Equatorial Guinea that was implemented as part of the national COVID-19 response and monitoring activities. We report how three distinct SARS-CoV-2 variants have dominated the epidemiological situation in Equatorial Guinea since March 2020. In addition, a case of co-infection of two SARS-CoV-2 VOC, Beta and Delta, in a clinically asymptomatic and fully COVID-19 vaccinated man living in Equatorial Guinea is presented. To our knowledge, this is the first report of a person co-infected with Beta and Delta VOC globally. Rapid identification of co-infections is relevant since these might provide an opportunity for genetic recombination resulting in emergence of novel SARS-CoV-2 lineages with enhanced transmission or immune evasion potential. Whole genome sequencing of SARS-CoV-2 viruses has been widely used since the beginning of the COVID-19 pandemic to facilitate understanding of virus biology and epidemiology. The World Health Organization (WHO) recommends that countries ship at least 5% of their COVID-19 samples to reference sequencing laboratories or keep producing sequencing data if they have the capacity assays which detect relevant mutations associated with SARS-CoV-2 variants of concern (VOC) . Mutatioe family , 11.In this brief research report, we describe the experiences and lessons learned from the SARS-CoV-2 genomic surveillance program at the Public Health Laboratory on Bioko Island, which was implemented as part of the national COVID-19 response and monitoring activities in Equatorial Guinea. We describe our approach to identify efficiently and timely SARS-CoV-2 VOC and co-infections in Sub-Saharan Africa, which is highly neglected when it comes to global genomic surveillance activities.th 2021.The number of confirmed COVID-19 cases and SARS-CoV-2 whole genome sequences from Central-Africa were obtained through the data repositories of \u201cOur World in Data\u201d (OWD) and the n = 281) and/or by whole genome sequencing (n = 206). The spike gene mutation-specific RT-qPCR assays target three spike gene single nucleotide polymorphisms (SNPs) and one spike gene deletion (\u039469/70) associated with VOCs of SARS-CoV-2 infections were reported from the insular region (Bioko Island) and 23.8% from the continental region (R\u00edo Muni) . Since the first case was identified on March 16th 2020, the country has experienced three distinct epidemic waves characterized by an increase of COVID-19-related hospitalizations on April 20th 2021 and the second dose on May 5th 2021. This person had no history of clinically significant underlying medical conditions, did not report any clinical symptom during the entire period of virus infection and has reported no travel history. As part of our routine contact tracing and SARS-CoV-2 surveillance activities his sample was analyzed for the presence of three spike gene mutations associated with VOCs. The mutation-specific RT-qPCR results for the spike gene mutations are shown for L452R , we were able to generate whole genome sequencing data for further investigation. The timeline of this co-infection case is shown in or L452R , E484K can be found below: Sample collection and retrospective sequence analysis was conducted according to the guidelines of the Declaration of Helsinki and approved by the National Technical Committee for the Response and Monitoring of the Novel Coronavirus , which is charged with preventing, containing, controlling, tracking, and evaluating the development and evolution of COVID-19 in Equatorial Guinea. Informed consent for publishing the Beta and Delta VOC co-infection case was obtained from the patient. Publication of the SARS-CoV-2 epidemiological, genomic surveillance data and the co-infection case was additionally approved by the Equato-Guinean Ministry of Health and Social Welfare.TS and CD: conceptualization, supervision, and project administration. SH, MM, PW, SR, and TS: methodology. SH, PW, and TS: software and validation. SH and TS: formal analysis, data curation, and visualization. SH, MM, EN, NB, TO, MO, PW, and UV: investigation. DM, MA, and WP: resources. SH, CD, and TS: writing\u2014original draft preparation. All authors have read and agreed to the submitted version of the manuscript.The funding for this work was provided through the public\u2013private partnership, the Equatorial Guinea Malaria Vaccine Initiative, supported by the Government of Equatorial Guinea, Ministries of Mines and Hydrocarbons, and Health and Social Welfare, Marathon Equatorial Guinea Production Limited, Noble Energy, Atlantic Methanol, Production Company, and the Equatorial Guinea Liquefied Natural Gas Company. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Nature Communications 10.1038/s41467-021-25552-0, published online 02 September 2021.Correction to: This Article contained an error in the Acknowledgments section. In the Acknowledgements section the author omitted to include a grant from the Ministry of Science and Technology of China, 2020YFA0112302, for which the work received financial support.This error has been corrected in the HTML and PDF versions of the Article."} +{"text": "The chapter on mental disorders of the 11th revision of the International Classification of Diseases (ICD-11) has been now finalized. Training of mental health professionals in the use of the chapter is taking place worldwide.Information is provided on the ICD-11 training courses taking place recently, including that co-organized by the Naples World Health Organization (WHO) Collaborating Centre on Research and Training in Mental Health and the European Psychiatric Association; those which will be held in the next few months, such as the one co-organized by the World Psychiatric Association and the Global Mental Health Academy, to be held online from 8 to 29 November 2021; and the training course set up by the WHO Collaborating Centre on Mental Health at the Columbia University, in collaboration with the WHO Department of Mental Health and Substance Use, which can be accessed only by the members of the WHO Global Clinical Practice Network.Psychiatrists of all countries of the world are encouraged to become familiar with the ICD-11 chapter on mental disorders, which will be adopted shortly by most countries worldwide. The chapter on mental, behavioural and neurodevelopmental disorders of the 11th revision of the International Classification of Diseases (ICD-11), has now been completed. The structure of the chapter, the most important changes with respect to the 10th revision of the International Classification of Diseases, and the most significant differences from the Diagnostic and Statistical Manual of Mental Disorders, 5th edition, have been recently described in detail \u201317.Education and training of mental health professionals in the use of the ICD-11 chapter is now taking place worldwide, mostly focusing on psychiatrists and psychologists, under the supervision of a WHO International Advisory Group led by G.M. Reed. Training courses have been implemented within the 18th and 19th World Congresses of Psychiatry [In this manuscript, information is provided on the ICD-11 training courses taking place recently. In April 2021, the Naples World Health Organization (WHO) Collaborating Centre on Research and Training in Mental Health and the European Psychiatric Association organized an online 20-h training course, coordinated by G.M. Reed and M. Maj. The course, subdivided into four sessions, has covered schizophrenia and other primary psychotic disorders, disorders specifically associated with stress, mood disorders, anxiety and fear-related disorders, obsessive\u2013compulsive and related disorders, feeding and eating disorders, personality disorders, disorders due to substance use, disorders due to addictive behaviours, and compulsive sexual behaviour disorder. W. Gaebel, M. Cloitre, M. Maj, C.S. Kogan, P. Monteleone, M. Swales, J.B. Saunders and N.A. Fineberg composed the Faculty.www.wpanet.org). The Faculty will include W. Gaebel, M. Cloitre, M. Maj, C.S. Kogan, O. Gureje, M. Swales, J.B. Saunders and N.A. Fineberg. A training session covering psychotic disorders and mood disorders was organized by the psychiatric association of Turkey in June 2021. A similar event was organized by the UK Royal College of Psychiatrists in May 2021.A similar training course, co-organized by the World Psychiatric Association and the Global Mental Health Academy, will take place online from 8 to 29 November 2021 (see https://gcp.network). The course consists of 15 online training units, each focusing on a different group of disorders and taking from one hour to one hour and a half. Each unit provides a description of the relevant diagnostic group and the main innovations with respect to the ICD-10. The outcome of training is checked through knowledge questions. Participants have the opportunity to practice by applying diagnostic guidelines to clinical case examples. This training course is going to be available also in Spanish, and additional translations are planned.The WHO Collaborating Centre on Mental Health at the Columbia University, in collaboration with the WHO Department of Mental Health and Substance Use, has recently set up a training course with exclusive access to the members of the WHO Global Clinical Practice Network . Among them, about 51% are psychiatrists and 30% psychologists.All health professionals working in mental health or primary care are welcome to join the Network in order to become familiar with the ICD-11 chapter on mental disorders, which will be adopted shortly by most countries worldwide."} +{"text": "Objectives: Opioid dependence has been a threat to public health for hundreds of years. With the increasing number of studies on acupuncture-related therapies for opioid dependence patients receiving methadone maintenance treatment (MMT), its effect of acupuncture therapy in treating MMT patients remains controversial. Therefore, we conducted a multiple-treatments meta-analysis, and incorporated both direct and indirect comparisons, in order to discover the most effective treatment for opioid dependence patients receiving MMT.Methods: Five English databases and three Chinese databases were searched from its inception to August 20, 2020, in order to compare the effects of acupuncture-related therapies and MMT, which was summarized as Western medicine (WM) in the following texts. The quality of studies was assessed according to Cochrane's risk of bias tool 5.1.0, and a pair-wise meta-analysis, cumulative meta-analysis, and the network meta-analysis was performed using the R software (Version 3.6.1) and STATA (Version 14.0). The primary outcome was the effective rate, which was calculated by the ratio of detoxifying patients to the total. The secondary outcome was the Modified Himmelsbach Opiate Withdrawal Scale (MHOWS).Results: A total of 20 trials were included, which consisted of comparisons among WM, traditional Chinese medicine (TCM), and the four types of acupuncture, namely, manual acupuncture (MA), electro-acupuncture (EA), auricular acupuncture (AA), and transcutaneous electrical acupoint stimulation (TEAS). Though none of the trials were at low risk of bias. In the pair-wise meta-analysis, no statistically significant differences were observed in terms of the effective rate. Furthermore, MA was more efficacious than WM, EA, and TEAS in MHOWS, with mean differences (MDs) of , , and , respectively. In the network meta-analysis, MA was more effective than WM on the effective rate, and on MHOWS. TEAS was more effective than WM on MHOWS. Synthetically, MA had the highest probability to rank first in treating opioid dependence.Conclusions: The existing evidence shows that acupuncture related-therapies may effectively be used for treating patients receiving MMT, and that manual acupuncture may be the best choice for opioid dependence among all kinds of acupuncture-related therapies. Nevertheless, reducing the relapse and promoting the recovery of opioid dependence need more efforts from not only the medical industry but also government support, security system, and educational popularization. To strengthen the assurance of acupuncture-related therapies in the treatment of opioid dependence, we expected that clinical trials with high quality would be conducted, to provide more confident evidence. According to the World Drug Report 2018, ~31 million people are affected by opioid-use disorders, which cause the greatest burden of severe disease and drug-related deaths worldwide. In China, ~2.5 million people use illicit drugs, and opioid and methamphetamine represent the majority. In order to solve this prominent public health problem, methadone has been widely used to suppress withdrawal symptoms induced by the abrupt discontinuation of drugs, decrease the use of opioids and other illicit drugs, and reduce criminal activities. At present, opioid agonist treatment has effectively reduced the incidence of HIV and hepatitis C caused by sharing needles , especially during the first several weeks of methadone stabilization. Patients often complain of insomnia and cravings during MMT, which may affect patient compliance with MMT and contribute to the risk of relapse. Furthermore, as an artificial opioid compound, the long-term use of methadone can impair patients' cognitive function and sustained attention and reduce the striatal dopamine transporter function. Although the use of MMT prolongs the dependence period, this may create new drug dependence at the same time . Hence, A systematic review was conducted following the general principles outlined in the Center for Reviews and Dissemination (CRD) Guidance and the PRISMA statement. We registered this systematic review in the PROSPERO database (CRD: 42021233950).We searched 8 professional digital databases from their inception to August 2020: Medline, EMBASE, Cochrane Library, SCOPUS, Web of Science, CNKI, VIP, and Wanfang. A search term and search strategy were developed for each database published in the Chinese or English language, and the effects of different kinds of acupuncture therapies in opioid dependence were evaluated.We intended to include adult participants. However, it was estimated that opioid dependent patients were emerging in a younger population. Hence, we adjusted the lower bound of age to 16 years old. These enrolled participants were diagnosed with opioid dependence according to the Chinese Classification and Diagnostic Criteria of Mental Disorders (CCMD) or Diagnostic and Statistical Manual of Mental Disorders (DSM). Based on the diagnosis criterion, participants were recruited when they had the following: (1) opioid abuse history; (2) drug dependent syndrome, withdrawal symptoms, or personality change after taking opioids; 3) impaired social function or even criminal behaviors due to drug-taking; and (4) mental health disturbance caused by opioid recovery rate or effective rate, assessed by the quantity of participants, who were completely detoxified, nearly detoxified, and partially detoxified from the therapy; (2) withdrawal symptoms scores measured by the Modified Himmelsbach Opiate Withdrawal Scale (MHOWS) . In effeIn order to evaluate the methodological quality of included studies, two reviewers (S.G. and W.L.) independently assessed the risk of bias according to Cochrane's risk of bias tool 5.1.0 . There wTwo reviewers (X.W. and Y.D.) screened the full texts, extracted the correlated information from all the included studies, and cross-checked the information, in order to ensure the consistency and accuracy of the extraction. We extracted the general information , drug-abuse circumstances , and information of interventions . The information above was concluded in a study characteristics table.A head-to-head meta-analysis was performed to evaluate the efficacy of acupuncture-related therapies and opioid substitutions in opioid dependence. The quality of studies was assessed according to Cochrane's risk of bias tool 5.1.0. We used R software (Version 3.6.1) to conduct the pair-wise meta-analyses and cumulative meta-analyses. The meta packages were used for pair-wise meta-analyses with random-effects model command, \u201cmetabin\u201d command for dichotomous data while \u201cmetacont\u201d command for continuous data. The effective rate was reported as risk ratio (RR), and for continuous data, the MHOWS was reported in mean difference (MD). All reported data corresponded with the 95% confidence intervals (CIs). The cumulative meta-analyses were carried out using \u201cmetacum\u201d command, verifying the results of meta-analyses.Two computer software, R software (Version 3.6.1) and STATWe constructed a fixed-effect model and a random-effect model, and set the parameter values as follows: n.chain = 3, factor = 2.5, n.adapt = 5,000, and n.iter = 20,000. Three chains yielded 20,000 iterations and the factor of 2.5.Based on the deviance information criterion (DIC) from R and the diagnostic parameters potential scale reduction factor (PSRF) from Brooks-Gelman-Rubin, we judged the comparison between the fixed-effect model and random-effect model. If the PSRF value was close to 1 and the DIC value was lesser, the convergence of the model was more suitable.P-value of the node-splitting analysis was >0.05, the direct evidence was consistent with the indirect evidence. Inconsistency was defined as the differentiation between the direct and indirect evidence, with a P < 0.05. These analyses were performed after the derivation of the inconsistency was determined ; (2) incomplete data (n = 36), ineligible outcomes (n = 85), interventions (n = 102), and participants (n = 45). The flowchart for the trial selection is shown in A total of 2,154 results were collected from eight digital databases using the search strategy. A total of 411 applicable full-text results were evaluated for eligibility, and 391 records were excluded, based on the following: (1) non-randomized studies as an opioid-substitution were summarized as Western medicine (WM) groups. The Chinese medical formulae, Chinese formulated products, and Chinese herbal detoxification capsules were summarized as traditional Chinese medicine (TCM) groups. Since MMT is a basic treatment for opioid dependence, we omitted MMT in the acupuncture-related therapy groups.Ultimately, 1,661 participants in 16 two-arm studies and 336 participants in 4 three-arm studies were included for the network meta-analysis. In the two-arm studies, 3 of 16 trials compared manual acupuncture (MA) and Western medicine (WM) , 16, 27,For the outcome measures, 11 trials reported the effective rate , 31\u201334, P-value to < 0.05. In order to analyze the effect sizes, risk ratios (RRs) were used for the effective rate and mean difference (MD) was used for the MHOWS score. The forest plots are shown in the In order to measure the efficacy of these six interventions, a classic meta-analysis was performed using the random-effects model, in order to compound studies with the same interventions. We considered a presence of statistically significant difference by setting the P = 0.34), AA , EA , and TCM had a higher effective rate, when compared to WM. Furthermore, TEAS was less efficacious than WM. For the comparisons among the other four acupuncture-related therapies, only one study compared and reported the data . However, no statistical difference was found in the above comparisons.A total of 14 studies that covered 8 head-to-head comparisons reported the effective rate . AccordiFor the MHOWS, we judged lower scores of MHOWS as less pertinent symptoms of participants. Nine studies that covered eight direct comparisons reported the effects of five interventions .P < 0.01). TCM , EA , and TEAS were better in decreasing the MHOWS score when compared to WM; however, no statistical differences were found in the comparisons. For the direct comparisons among acupuncture-related therapies, MA was more efficacious than EA and TEAS . In comparing with TCM, EA and MA had better effects on the MHOWS score, although no statistical difference was found in the above comparisons.A significant decrease in MHOWS score was observed in MA comparing to WM , AA vs. WM , and MA vs. WM , and the MHOWS in comparisons of EA vs. WM , MA vs. WM , and TCM vs. WM were supportive of the results in meta-analyses. The forest plots were in the We conducted a network meta-analysis to exhaustively compare and rank the different interventions for opioid dependence. The network plots for the effective rate and MHOWS are presented in P-values between the direct and indirect effects were >0.05. The PSRF value close to 1 indicated that the convergence of the model was more suitable, and that the result was stable and convincing.We used node-splitting analysis to assess the consistency, and all The network meta-analysis for the effective rate is presented in In the results of the network meta-analysis for the MHOWS , all acuP-value of > 0.05. The node-split plots revealed that there was barely a heterogeneity or inconsistency between the direct and indirect evidence in each comparison.Node-split was conducted to detect the inconsistency and heterogeneity for the effective rate and MHOWS score , 10. TheTo date, the present study is the first to conduct a network meta-analysis on various therapies for opioid dependence patients receiving MMT. This network meta-analysis would help to build up connections between individual interventions, evaluate the efficacy of all therapies, and conclude the best treatment by ranking these therapies. From the results of the present network meta-analysis and the rank of the interventions, and among all the acupuncture-related therapies, manual acupuncture has a prominent effect on the effective rate and MHOWS score. Furthermore, both the direct evidence in the meta-analysis and indirect evidence made by Bayesian statistical methods indicated that manual acupuncture may effectively allay the withdrawal symptoms, and help patients get rid of opioid dependence. Some studies demonstrAt present, many patients desire to completely throw off addiction and live in a normal way without taking methadone. However, for the patients who are used to their daily dosage, it is difficult to change their dosage. To avoid withdrawal syndrome caused by reducing methadone, most of them choose to take the same dosage, even for the rest of their lives. The most reason we highly recommend acupuncture for MMT patients is that acupuncture, as the adjunctive therapy, can largely improve the phenomenon, helping patients get rid of opioids by controlling withdrawal syndrome. Acupuncture is now being used in many diseases , the safCompared to the former meta-analysis, one of which focused on the effect of acupuncture combined with opioid receptor agonists in treating opiate-withdrawal symptoms, the present study included and reported in detail the different types of acupuncture and ranked the possible best intervention for opioid dependence. Another familiar direction is the comparison of the dose response and efficacy among opiate maintenance treatment, burenorphine, and other pharmacological adjunctive interventions \u201343. The Although the present network meta-analysis strongly persuades that acupuncture-related therapies are more effective than WM, there were some limitations of the present study. First, the RCTs were published at least 15 years ago which commonly had no rigorous study design, or a detailed description on randomization, allocation concealment, or blinding. Therefore, the low quality of the included trials may downgrade the confidence on the recommendation. Second, the two outcome measures were both correlated with the MHOWS score, which indicates that the efficacy was only judged by the decreasing scores. In order to determine the independent effect of different types of acupuncture therapies in the future, the daily reduction of methadone while receiving acupuncture combined with MMT can be measured, which may more directly contribute to a positive effect in opioid dependence. Last, the relapse of patients varied from person to person, that is, some of the patients were at their first abstinence, while some of the patients may have undergone this twice or had more relapses. This may affect the present results in a way. If possible, researchers can collect more information and explore the interior connection between relapse and detoxification.The existing evidence shows that acupuncture related-therapies may effectively be used for treating patients receiving MMT, and the results of this network meta-analysis support manual acupuncture may be the best choice for opioid dependence among all kinds of acupuncture-related therapies. Nevertheless, reducing the relapse and promoting the recovery of opioid dependence needs more efforts from not only the medical industry but also government support, security system, and educational popularization. To strengthen the assurance of acupuncture-related therapies in the treatment of opioid dependence, we expected that clinical trials with high quality would be conducted, to provide more confident evidence.The original contributions presented in the study are included in the article/HW and LL designed the search strategy. SG and WL performed the literature search. HW and RC screened the studies for eligibility and wrote the first draft of the manuscript. RC, YD, and XW performed the data extractions. HW, RC, and PZ conducted the statistical analyses. LL, SX, and YZ were responsible for the manuscript editing and review of the manuscript.This work was supported by the National Natural Science Foundation of China (82174527), the special project of Lingnan modernization of traditional Chinese medicine in 2019 Guangdong Provincial R & D Program (2020B1111100008), the High-level university construction of GZUCM (A1-2601-21-415-024), the National Natural Science Foundation of China (81872261 to SX); the Scientific Project of Guangzhou (201803010013 to SX), and the Luo Yongjia Famous Traditional Chinese Medicine Studio, Guangdong Traditional Chinese Medicine Office ([2019] No. 5 to YZ).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "These relationships were also significant in the gender-specific analysis. Thus, the yield of not-for-sale fruits and vegetables might contribute to the intake of fruits and vegetables in Japan.This study investigated the relationship between prefecture-level yield of not-for-sale fruits and vegetables and individual-level fruit and vegetable intake in Japan. Data were drawn from the Japanese National Health and Nutrition Survey and National Crop Survey of 2016. Random intercept models were used for the analyses. Individual-level fruit and vegetable intake was used for the dependent variable, and prefecture-level yield of not-for-sale fruits and vegetables was used for the independent variable as a fixed effect. In addition, participants\u2019 characteristics and health-related factors at the individual level were also put into independent variables as fixed effects. The prefectures were used as random intercepts. It was found that prefecture-level yield of not-for-sale fruits and vegetables was significantly related to individual-level fruit and vegetable intake (vegetable: B = 0.390, The World Health Organization (WHO) and the UN Food and Agriculture Organization (FAO) promote increased consumption of fruits and vegetables because it can reduce the risk of certain noncommunicable diseases and the mortality these diseases entail . SeveralCertain environmental factors have been known to influence fruit and vegetable intake . A systeThe influence of the cultivation of fruits and vegetables with fruit and vegetable intake has been widely reported in many previous studies ,14,15,16In Japan, it has been found that obtaining nonmarket food both from within and outside of the household could affect dietary intake ,22,23,24Thus, it was hypothesized that residents in areas with high yields of not-for-sale fruits and vegetables would have high intakes of fruits and vegetables. The mechanism would run as follows: the harvested not-for-sale fruits and vegetables are first consumed in the grower\u2019s home. Then, surplus fruits and vegetables are provided to their neighbors. In addition, some neighbors who have received more than they can use may provide their excess to their other neighbors. In an ecosystem of this type, residents in areas with high yields of not-for-sale fruits and vegetables are likely to enjoy high intakes of fruits and vegetables.This study investigated the relationship between prefecture-level yield of not-for-sale fruits and vegetables and individual-level fruit and vegetable intake in Japan. The yields of the fruits and vegetables used in this study were taken to be the amount recorded in the Japanese national statistics. This figure does not include fruits and vegetables produced in a home garden or a community garden. However, previous studies in Japan have found that 96% of farmers grow vegetables for their own consumption and 84% give away the vegetables they grow to their neighbors . In addiThis was a cross-sectional study that used data from two Japanese national surveys.One of these was the National Health and Nutrition Survey (NHNS) 2016 . That suThe other major source of data was the National Crop Survey (NCS) 2016 ,29. ThisThis study used individual-level data on fruit and vegetable intake of NHNS as the dependent variables. Fruit and vegetable intake was provided and used for analyses. In addition, the total fruit and vegetable intake was calculated and used for the analyses. The unit used for all analyses was daily intakes (g) per participant.The survey method for food and nutrition intake in NHNS was as follows: food and nutrition intake was determined by the dietary record, using a weighted method . In addi6, it was converted to the grams per person per day for the analysis.This study used prefecture-level yield of not-for-sale fruits and vegetables of the NCS as independent variables. The yield of not-for-sale fruits and vegetables for each prefecture was calculated by subtracting the shipment amount from the yield using data for the yield and shipment amount for each prefecture from the NCS. The shipment and yield of the NCS were aggregated by annual tonnage for each prefecture ,29. TherNCS 2016 (vegetables and fruits) provides statistics on 55 major items produced in Japan ,29. Howe2). Energy intake was calculated from a survey of food and nutrition intake already written [Prefecture, gender, age, lifestyle, drinking habits, smoking habits, body mass index (BMI), and energy intake were used for the analyses. These data were individual-level data provided by NHNS . Gender, written . These vFrom the data on 26,354 individuals obtained from the National Health and Nutrition Survey, excluding those from participants aged younger than 20 and more than 79, those pregnant and lactating, and those with missing data in the items used for analysis, data from 15,046 participants aged 20\u201379 years were used in this analyses. First, in the preliminary analyses, the yields of not-for-sale fruits and vegetables and the intake of fruits and vegetables were calculated for each prefecture, and Spearman\u2019s correlation coefficient between these variables was calculated.Then, random intercept models were used for the main analyses . The prep < 0.05.All analyses were conducted using IBM SPSS Statistics for Windows, version 23.0; IBM Japan, Ltd.: Tokyo, Japan. Statistical significance was set at The NHNS data were obtained with permission from the Ministry of Health, Labor and Welfare. The NCS data were obtained from the Japanese official statistics portal site e-stat. The data did not contain any personally identifiable information. This study was approved by Takasaki University of Health and Welfare Research Ethics Committee .p-Value) between intake and yield among prefectures were 0.365 (0.010), 0.442 (0.002), and 0.401 (0.006) for vegetables, fruits, and fruits and vegetables, respectively.In p values of null models are shown in In p-value for trend of categories was less than 0.001. Similar significant associations were found when analyzed by gender. The null model\u2019s intercepts for all participants (p < 0.001), men (p = 0.010), and women (p = 0.001) were significant. For all participants, men, and women, the AICs were smaller when the individual-level variables were input from null models. The AICs were even smaller in the models that input the yield at the prefecture level.For vegetables, the regression coefficient (B) was 0.390 when the yield of not-for-sale vegetables was used as an interval scale. Similarly, there was a significant association in the quartile categories . The p-value for trend of categories was 0.013. Similar significant associations were found for analysis by gender instead of quartile categories for women. The null model\u2019s intercepts were significant in all participants (p = 0.001), men (p = 0.042), and women (p = 0.007). The AICs were smaller when the individual-level variables were input than null models for all participants, men, and women. For the models that input the yield at the prefecture level, the AICs were even smaller compared with the other models.For fruits, the B was 0.268 (95% CI: 0.099 to 0.438) when the yield of not-for-sale fruit was used as an interval scale. Similarly, there was a significant association in the quartile categories . The p-value for the trend of categories was 0.001. Similar significant associations were found when analyzed by gender. The intercepts of null models of all participants (p < 0.001), men (p = 0.005), and women (p = 0.001) were significant. For all participants, men, and women, the AICs decreased in the order of null models, individual-level models, and prefecture-level models. For fruits and vegetables, B was 0.357 (95% CI: 0.167 to 0.548) when the yield of not-for-sale fruits and vegetables was used as an interval scale. Similarly, there was a significant association in the quartile categories . The This study investigated the relationship between prefecture-level yield of not-for-sale fruits and vegetables and individual-level fruit and vegetable intake in Japan. As a result, the positive relationship between prefecture-level yield of not-for-sale fruits and vegetables and individual-level fruit and vegetable intake was found. To date, no studies have demonstrated such an association. This is the first study to identify the yield of not-for-sale fruits and vegetables as an environmental factor related to the intake of fruits and vegetables, and it may contribute to the development of a health-promoting food environment.In preliminary analyses, it was found that there is a prefectural-level positive correlation between the yield of not-for-sale fruits and vegetables and fruit and vegetable intake in Japan. A previous study has reported a correlation of this type for vegetables, but it did not examine fruits or the total amount of fruits and vegetables . This stWe found a positive relationship between prefecture-level yield of not-for-sale fruits and vegetables and individual-level fruit and vegetable intake. Many positive relationships between fruit and vegetable cultivation/receiving and fruit and vegetable intake have been reported ,22,23,24Food access is positively associated with the consumption of fresh foods, such as vegetables and fruits . HoweverThis study had several limitations. First, since it was conducted in Japan, its results may only pertain to that country and not to other countries or regions. In Japan, it is common to share one\u2019s produce surplus to neighbors, so the results of this study were obtained. Therefore, results of similar studies will be different in countries and regions where there is no accepted culture of passing on one\u2019s surplus to others. In this study, only the yield of not-for-sale fruits and vegetables produced by farmers were used as independent variables. It should be noted that the crops produced by nonfarmers in home gardens and community gardens were not included. This study also examined data at the prefecture level. However, the actual transfer of surplus often appears within smaller regional units. Finally, this was a cross-sectional study. Therefore, the causal relationship of the association confirmed in this study cannot be confirmed.This study revealed the positive relationship between the prefectural-level yield of not-for-sale fruits and vegetables and individual-level fruit and vegetable intake. An environment where not-for-sale fruits and vegetables could be harvested abundantly might be contributing to the resident\u2019s intake of abundant fruits and vegetables in Japan."} +{"text": "Cell Death & Differentiation 10.1038/s41418-021-00752-9Correction to: The original version of this article unfortunately contained a mistake in the affiliations. Since the publication of this article, the authors have noticed an error in the affiliation of the first author of the manuscript, Dr. Josephin Koschel. The correct affiliations are as follows:Josephin Koschel 1, 51 Institute of Medical Microbiology and Hospital Epidemiology, Hannover Medical School, 30625 Hannover, Germany5 Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, 39120 Magdeburg, GermanyThe authors apologize for the mistake. The original article has been corrected."} +{"text": "Sinobdella sinensis (Synbranchiformes: Mastacembelidae) from China's Qiantang River for the first time. The mitochondrial genome of S. sinensis was sequenced to be 16,543\u2009bp in length, larger than S. sinensis from China's Yangtze River. The genome contains 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs, two central non-coding regions (the control region and the origin of light strand replication), identical to most other vertebrates. Phylogenetic analysis highly supported that S. sinensis from China's Qiantang River was different from other Mastacembelus fish. However, it showed a close relationship with Macrognathus aculeatus. These data would explain the evolutionary mechanisms and biogeography of the family Mastacembelidae and is helpful for the conservation of genetics and stock evaluation for S. sinensis.This study determined the mitochondrial genome of Sinobdella sinensis (Bleeker 1870) is an East Asian species of the family Mastacembelidae of the order Synbranchiformes, the only species in the genus Sinobdella according to FishBase , and deposited at the National Original Breeding Farm of black Amur bream from China's Qiantang River . The total genomic DNA from the fin tissue (assigned as ZHCQ202103) was extracted by the phenol\u2013chloroform extraction method except for ND6 and eight tRNA, which are encoded on the light strand (L-strand). Within the genome, all the 13 PCGs included the orthodox start codon ATG except for CO1, which is initiated with GTG. However, the stop codons of the 13 PCGs differ, these terminating with TAG, TAA, TA\u2013, or T\u2013. The origin of light strand replication (OL), which extends up to 31 nucleotides, is identified in the WANCY region. The second non-coding region, the control region (D-loop), is located between the tRNA-Pro and tRNA-Phe with 919\u2009bp in length. The phylogenetic tree of S. sinensis is shown in S. sinensis from China's Qiantang River is different from other Mastacembelus fish. However, S. sinensis from China's Qiantang River is not initially grouped with S. sinensis from China's Yangtze River but clustered first with Macrognathus aculeatus from China's Pearl River. SH-aLRT is a fast nonparametric version of an approximate likelihood ratio test (aLRT) developed and implemented in the PHYML phylogenetic inference software (Anisimova et al. S. sinensis MZ188892, M. aculeatus KT443991, and M. aculeatus KF636363 are higher than 80%, which means the credibility of the SH-aLRT test is high (Hoang et al. Macrognathus spp. and Sinobdella spp., although there are many similarities in morphology.The complete mitochondrial genome of"} +{"text": "Social media has been crucial for seeking and communicating COVID-19 information. However, social media has also promulgated misinformation, which is particularly concerning among Asian Americans who may rely on in-language information and utilize social media platforms to connect to Asia-based networks. There is limited literature examining social media use for COVID-19 information and the subsequent impact of misinformation on health behaviors among Asian Americans. This perspective reviews recent research, news, and gray literature to examine the dissemination of COVID-19 misinformation on social media platforms to Chinese, Korean, Vietnamese, and South Asian Americans. We discuss the linkage of COVID-19 misinformation to health behaviors, with emphasis on COVID-19 vaccine misinformation and vaccine decision-making in Asian American communities. We then discuss community- and research-driven responses to investigate misinformation during the pandemic. Lastly, we propose recommendations to mitigate misinformation and address the COVID-19 infodemic among Asian Americans. During public health emergencies like the 2003 SARS pandemic, 2009 H1N1 swine flu pandemic, 2016 Ebola epidemic, and current COVID-19 pandemic , the intAsian American social networks are distinct from other communities. These networks may include family in other Asian countries and include dissemination of non-English information on social media. However, there is a paucity in literature examining use of social media for COVID-19 information among Asian Americans, types of misinformation disseminated on social media in these networks, and subsequently the impact of such information on Asian Americans' health behaviors. We conducted a narrative review to examine the use of social media platforms to disseminate COVID-19 misinformation across Chinese, Korean, Vietnamese, and South Asian Americans, four of the largest Asian subgroups in the United States (U.S.). We discuss the linkage of COVID-19 misinformation to health behaviors, with emphasis on misinformation about COVID-19 vaccines and vaccination confidence, as well as community-driven responses to address COVID-19 misinformation among Asian Americans. Lastly, we propose recommendations to mitigate misinformation and address the infodemic in Asian American communities.Health misinformation is prevalent across various social media platforms. A systematic review of 69 studies identified Twitter, YouTube, and Facebook as the most common social media platforms for health information dissemination, while less popular platforms included WhatsApp, Pinterest, Tumblr, BK, and Myspace . There aResearchers reported that social media was nationally the third most common information source for COVID-19 information and misinformation . SimilarAsian Americans, who comprise 5.5 percent of the U.S. population , are leaDespite the prevalent use of social media among Asian Americans, there has been limited research on Asian Americans' pattern of social media use by age, gender, and socioeconomic status . FrequenAmidst COVID-19-related uncertainty and fears, Asian Americans are turning to the internet and social media for the latest updates on the pandemic, such as confirmed COVID-19 cases, government-issued COVID-19 policies and guidelines, and COVID-19-related health information , COVID-19 symptoms, diagnosis, and treatment) , 31, whiExtant literature on social media, Asian Americans, and COVID-19 has focused on how contentious COVID-19 misinformation contributed to the increase in discrimination, racism, and violence against Asian Americans , 34, as In response to alarming COVID-19 misinformation in Asian American communities, community and public health leaders are spearheading efforts to combat the impacts of misinformation and to disseminate scientific COVID-19 information. In New York City, in-language virtual webinars were held by the NYU Center for the Study of Asian American Health (CSAAH), in partnership with community and academic partners, for Chinese, Filipino, South Asian, Arab American, and Hispanic communities, to answer questions and concerns about COVID-19. Likewise, the Filipino Young Leaders Program (FYLPRO) developed the Caretaker Program, Tayo, a virtual help desk that provides accurate and timely information on COVID-19 to the Filipino community in Tagalog and English . SimilarNationally, the Progressive Vietnamese American Organization (PIVOT) launched the Viet Fact Check Project to release in-language information on COVID-19 vaccination, to dispel common myths, and to encourage higher uptakes of vaccination in the Vietnamese community . The VieMisinformation, especially COVID-19 misinformation, has far-reaching impacts on health and wellbeing for Asian Americans and other minority groups that face existing, unique health disparities , 13, 56.Identification and correction of misinformation across social media is not enough to end the COVID-19 infodemic. In concordance with the deficit hypothesis , public In addition, the public health response can utilize and leverage community health workers (CHWs), who are vital members representative of the community, to identify COVID-19 misinformation and to inform community members of ways to be aware of misinformation and provide accurate in-language information.Furthermore, many forms of online misinformation, particularly conspiracy theories, are due to complex psychological mechanisms that need to be further examined to thoroughly combat and contain the infodemic in Asian American and other communities. People are also more likely to have strong conspiracy beliefs that undermine any scientific guidance when they are experiencing high levels of anxiety and uncertainty . ResearcSocial media has inevitably promulgated the transnational exchange of COVID-19 misinformation during the pandemic, prompting academic and community action to combat COVID-19 misinformation on social media. We speculate that Asian Americans are likely to utilize social media to receive COVID-19-related health information because they are able to effectively communicate with family and friends, particularly due to the dearth of COVID-19 information by the federal government in diverse Asian languages. We recommend multi-sectoral efforts between public health officials, community leaders, researchers, and social media companies to identify and mitigate health misinformation on social media, as well as the inclusion of CHWs to educate community members on misinformation and provide accurate scientific information. Improving eHealth and digital literacy among Asian Americans could also aid in the evaluation of identifying misinformation and seeking trustworthy and reputable websites. More research on the psychological mechanisms behind conspiracy theories and misinformation could led to effective health promotion communication strategies and interventions that improve acceptance of scientific evidence and public health guidance needed during public health emergencies.Research is also needed to address the origins and mediators of susceptibility to COVID-19 misinformation. For example, the exploration of how social media platforms, usage patterns, and usage of social media for health information by disaggregated Asian American subgroups in the U.S., as well as by other racial and ethnic groups is needed. Increase research efforts are necessary to quantitatively and qualitatively understand the social, economic, and health impacts of COVID-19 misinformation in Asian American communities. The consequences of misinformation on health behaviors in Asian American communities will likely span beyond the impacts of the COVID-19 infection and will require culturally- and linguistically-concordant interventions to address health disparities.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.SC, SA, SY, and SK conceptualized the manuscript. SC and SA wrote the first draft of the manuscript. L\u00d0, SY, and SK provided critical review of the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version.This work was supported by the National Institutes of Health (NIH), National Institute on Minority Health and Health Disparities (NIMHD) Award Number U54MD000538, and the preparation of this manuscript was supported in part by U.S. Department of Health and Human Services, Centers for Disease Control and Prevention (CDC) Award Numbers NU38OT2020001477, CFDA number 93.421, 1NH23IP922639-01-00, CFDA number 93.185, and NIH NHLBI Community Engagement Alliance (CEAL) Non-Federal 1OT2HL156812-01, Westat Sub-OTA No: 6793-02-S013. L\u00d0 was supported by the NIH Resource Centers for Minority Aging Research (RCMAR) Award Number 5P30AG059302. The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of the funders.The contents of this publication are solely the responsibility of the authors and do not necessarily represent the official views of the funders.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "The coronavirus disease (COVID-19) pandemic highlighted that managing health emergencies requires more than an effective health response, but that operationalizing a whole-of-society approach is challenging. The World Health Organization (WHO), as the lead agency for health within the United Nations (UN), led the UN response at the global level through the Crisis Management Team, and at the country level through the UN Country Teams (UNCTs) in accordance with its mandate. Three case studies\u2014Mali, Cox's Bazar in Bangladesh, and Uzbekistan\u2014provide examples of how WHO contributed to the whole-of-society response for COVID-19 at the country level. Interviews with WHO staff, supplemented by internal and external published reports, highlighted that the action of WHO comprised technical expertise to ensure an effective whole-of-society response and to minimize social disruption, including those affecting peacekeeping in Mali, livelihood sectors in Cox's Bazar, and the education sector in Uzbekistan. Leveraging local level volunteers from various sectors led to both a stronger public health response and the continuation of other sectoral work. Risk communication and community engagement (RCCE) emerged as a key theme for UN engagement at country level. These collective efforts of operationalizing whole-of-society response at the country level need to continue for the COVID-19 response, but also in preparedness for other health and non-health emergencies. Building resilience for future emergencies requires developing and exercising multi-sectoral preparedness plans and benefits from collective UN support to countries. Coronavirus disease had many impacts outside of health, and therefore emergency preparedness needs to occur outside of health too. Managing the health risks and reducing the socioeconomic impact of emergencies requires a whole-of-society approach , definedEarly in the pandemic, WHO recognized that the impact of COVID-19 would expand beyond the health sector requiring a coordinated whole-of-society approach. In February 2020, the WHO Director-General requested activation of the United Nations (UN) Crisis Management Policy by the UN Secretary-General\u2014the highest possible level of crisis alert in the UN system , the COVMulti-sectorial collaboration during health emergencies has been recognized as an essential, and yet challenging, component of the response. The independent Review Committee on the Functioning of the International Health Regulations, in their review of the COVID-19 response, highlighted the need for multisectoral collaboration, and for WHO to work with Member States to engage stakeholders beyond the health sector to identify and address country level gaps in preparedness . The GloWorld Health Organization, as the lead agency for health within the UN, and in accordance with its mandate , provideThe three case studies were selected according to the following criteria: (a) different contexts including low- and middle-income countries and fragile settings with vulnerable populations, (b) different geographic regions, (c) large UN presence of 15 or more agencies, (d) agreement by the WHO Representative for interview. A convenience sample were selected from the collation of 70 case studies reported on in 2020 focusing on WHO's country level work in response to COVID-19 to proviMali is a low-income country with a long-term UN peace-keeper presence. Cox's Bazar in Bangladesh comprises a large vulnerable population of refugees in a low-to-middle-income country with an international humanitarian response. Uzbekistan is a middle-income country with a UN interagency presence where the UN is a development partner providing technical assistance. The WHO Representative or staff from each was interviewed about the role of WHO within the UN system in the whole-of-society response for COVID-19. The information gathered from the interviews is supplemented by internal and published reports.Mali is a fragile state in Africa with a large UN presence, consisting of 21 UN entities plus the Multidimensional Integrated Stabilization Mission in Mali (MINUSMA), established by the UN Security Council in 2013. Collectively, the UN in Mali comprises almost 19,000 staff and personnel, including 13,000 military, 5,600 civilians, and 1,560 employees of agencies . After dPrior to the COVID-19 pandemic, although there were many joint UN projects, most UN agencies operated within their own functions and mission. This changed for the COVID-19 response\u2014as the full machinery of the UN operated \u201cas one\u201d in an unprecedented way . The NatWorld Health Organization provided the link between the government and the UN Resident Coordinator for the COVID-19 response, based on their well-established relationship with the Ministry of Health and Social Development (MHSD). World Health Organization mobilized staff from other UN agencies to provide technical support to the government across whole-of-society activities. Of the 25 UN team members, seven were from non-WHO UN agencies including from UN Children's Fund (UNICEF) for risk communication and community engagement (RCCE), the UN Development Programme (UNDP) for program management and the World Food Program (WFP) for logistics and supply chain management. This structure meant that for the first time the National Health Authority collaborated with multiple UN agencies.The enabling role of WHO is illustrated by the public health and social measures adopted by MINUSMA to overcome the challenges in operating within COVID-19 restrictions. Guided by WHO recommendations and working closely with national authorities, MINUSMA implemented quarantine, body temperature checks before accessing camps, social distancing, mask wearing, and COVID-19 pre-deployment training to ensure its continued operation . MultidiGetting appropriate RCCE messaging to hard-to-reach populations was a challenge in Mali that was overcome by a whole-of-society approach supported by WHO . The UN Cox's Bazar, in Bangladesh, comprises a large refugee population spread across 34 highly congested subcamps along the Bangladesh/Myanmar border. The international response coordination in Cox's Bazar is managed under a situation specific mechanism led by a Strategic Executive Group at the national level and through the Inter Sector Coordination Group (ISCG) in Cox's Bazar. The ISCG comprised ten sectors and six inter sector working groups that provide humanitarian assistance to close to 900,000 Rohingya refugees . The WHOGuided by the WHO Emergency Response Framework , WHO BanPrior to the first COVID-19 cases in the district, a COVID-19 response plan was developed that outlined key activities across 11 identified thematic pillars of the national COVID-19 response plan . A dedicThe response to COVID-19 in Cox's Bazar provides many examples of how WHO supported whole-of-society COVID-19 activities. World Health Organization provided technical updates to the Heads of UN agencies and the ISCG, and between UN agencies, local community groups, donors/partners, and embassies to facilitate decision making, allocate resources, and mobilize funds. In collaboration with the technical working groups, the health sector developed an orientation package and online awareness training sessions for healthcare workers to ensure mainstreaming of cross-cutting themes including gender, protection from sexual exploitation and abuse, gender-based violence, protection, and child protection in the response. Prior to the reopening of the destination for domestic tourism, in collaboration with local government, WHO provided technical advice, training, and information materials on operating safely within the COVID-19 context to various sectors, including the tourism, agriculture, and livelihood sectors.One unique example of the role of WHO in the whole-of-society response to COVID-19 in Cox's Bazar was the provision of technical advice and coordination for repurposing Rohingya volunteers involved in garment manufacturing and tailoring as livelihoods activities to produce cloth face masks . World HRestriction to the camps was another challenge for the whole-of-society response as many volunteers from non-health sectors such as protection, education, and nutrition were unable to continue their work. To overcome this challenge, WHO utilized these volunteer networks in partnership with the other UN agencies for health activities, which also enabled their usual volunteer roles. The volunteers contributed to the community surveillance system established prior to COVID-19 , referreThe training provided to the volunteer networks enabled RCCE efforts as the volunteers disseminated these messages whilst completing their usual work . This inUzbekistan is a middle-income country in central Asia that has a UN interagency presence with a UNCT that comprises 24 UN agencies . World HA UN Crisis Management Team for COVID-19 was established and was co-chaired by WHO. The Crisis Management Team operated according to the national SPRP and the UN framework for the socio-economic response to COVID-19 in Uzbekistan. World Health Organization led the SPRP, contributed to the combined UN/WHO sitreps, was a co-chair of most COVID-19 related meetings, and a vital part of the UN decision making process. This further increase the visibility and added value of WHO within the UN system in Uzbekistan.There were six COVID-19 taskforces led by the UN and the government: capacity building, procurement, human rights and vulnerable groups, economic and social impact, education, and risk communication. World Health Organization led the capacity building taskforce and had a presence in all others. By MOH request, WHO was the liaison between the MOH and other UN agencies and the conduit between the COVID-19 taskforces. The Health Development Partnership meetings continued as the intersectoral communication and collaboration mechanism between the UN, non-UN partners, and government. World Health Organization presented technical information to each taskforce as needed and reviewed taskforce documents prior to distribution to the government\u2014a \u201ctranslator.\u201d The COVID-19 response cemented that intersectoral collaboration is crucial for advancing the health agenda with WHO as a key contributor.One example of WHO working across the whole-of-society response in Uzbekistan was in the education taskforce. Working closely with UNICEF and the UN Educational, Scientific, and Cultural Organization (UNESCO), WHO contributed to technical guidance on school closures, required school infrastructure, such as building ventilation and the safe reopening of schools. Information, education, and communication materials were developed in collaboration with the MOH, Ministry of Education, and Ministry of Pre-School Education with more than 6 million children in schools, 1.4 million children in pre-schools, 2 million parents received information, and 14,000 preschool institutions receiving materials .Online training mechanisms developed by WHO, in conjunction with the MOH Post Graduate Medical Institute, allowed experts to train large groups of healthcare workers in the public and private sector from across Uzbekistan (500 people per session) on COVID-19 related issues and topics, with the training materials made available to partners outside of the health sector.The COVID-19 pandemic has highlighted that responding to a global infectious disease pandemic has far-reaching implications outside the health sector that requires a whole-of-society approach. The UN, with its existing global presence, and guided by the three overarching global plans for health, humanitarian, and socio-economic response \u201312, was The COVID-19 pandemic has impacted everyone globally, in every facet of their lives. There has been a drastic decrease in the human development index globally , with maWorld Health Organization also provided leadership at the country level within the UN response globally, with 87% of WHO country offices reporting that they led the COVID-19 response within the UNCTs and 94% also reporting that their role within the UNCT expanded due to the pandemic . The thrRisk communication and community engagement has been highlighted as an integral component of the COVID-19 response that has been exacerbated by an \u201cInfodemic\u201d including misinformation and rumors . The thrThe COVID-19 pandemic has emphasized that managing health emergencies requires more than an effective health response, but that operationalizing a whole-of-society approach is challenging. Providing the actions and inputs from WHO only in this perspective article is a limitation which can be strengthened by further work to assess and present the perspectives of other UN agencies and beneficiaries in whole-of-society response. So that these experiences are not lost, WHO should continue the whole-of-society interactions within the COVID-19 response through the UN system, and in preparedness for other health and non-health emergencies. There are opportunities to sustain these efforts beyond the COVID-19 pandemic, such as cross-purposing volunteers within different sectors, optimizing use of digital technologies for RCCE, and strengthening platforms for online education to minimize the negative impact of future crises on children. Building resilience for future emergencies can also be achieved more broadly through developing and exercising multi-sectoral preparedness plans and through collective UN support to countries in prevention, risk governance, and forging critical partnerships. As stated in the SPRP, no single agency or organization can prepare for or respond to such an event on its own .The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.GS, MM, JE, OO, SA, and PG: conceptualization. GS, MM, JE, OO, J-PB, LK, KV, and MFS: data collection and interpretation. GS, MM, JE, OO, and KV: drafting the manuscript. GS, MM, JE, OO, J-PB, LK, KV, MFS, SA, and PG: review and revising manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."} +{"text": "Hematology Reports published its first volume in 2009, which included 10 publications: 8 original articles and 2 reviews [Hematology Reports became the official journal of the Society of Hematologic Oncology Italy . The journal publishes their scientific material from conferences and other events. The presidents of SOHO Italy are Dr. Claudio Cerchione, Prof. Dr. Giovanni Martinelli, and Prof. Hagop Kantarjian [In September 2019, Hematology Reports will be published by MDPI and will continue to serve the scientific community by publishing high-quality, open-access, peer-reviewed articles on all aspects of the prevention, diagnosis, and management of disorders of the blood. As of 2022, Hematology Reports will continue to publish high-quality and interesting research in the field of hematology.We would like to offer a warm welcome to all Editorial Board Members, SOHO society members, and authors. We hope"} +{"text": "Nature Communications 10.1038/s41467-021-25516-4, published online 15 September 2021.Correction to: The original version of this Article contained errors in the author affiliations.The affiliation of Malin Jonell and Beatrice Crona with Stockholm Resilience Centre, Stockholm University, Stockholm, Sweden was inadvertently omitted.The affiliation of Malin Jonell with Beijer Institute of Ecological Economics, The Royal Swedish Academy of Sciences, Stockholm, Sweden was inadvertently omitted.This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "The incidence of early-onset colorectal cancer (CRC), commonly defined as diagnosis prior to the age of 50\u2009years, has been increasing across the globe . In the In this issue of the Journal, Archambault and colleagues harness Overall, the following factors were associated with modestly increased risk of early-onset CRC: alcohol consumption of more than 28\u2009g/day or abstinence compared with 1-28\u2009g/day, high red meat intake (per sex- and study-specific quartile) compared with low intake , irregular nonsteroidal anti-inflammatory drugs use compared with regular use, and lower educational attainment compared with completion of at least college. The strengths of these associations were similar for late-onset CRC. As the increasing incidence of early-onset CRC was initially primarily driven by increases in rectal cancer . EEO is supported by T32 CA009621.Role of the funders: The funders had no role in the writing of this editorial or the decision to submit it for publication.Disclosures: The authors declare no conflicts of interest to disclose.Author contributions: Conceptualization, EEO and YC; Writing- original draft, EEO and YC; Writing- review and editing, EEO, YC, and GAC.Not applicable."} +{"text": "ABSTRACT IMPACT: Through its interdisciplinary, tailored approach, the FLOW program could change the way that we approach promoting healthy lifestyle changes in the primary care field. OBJECTIVES/GOALS: The goal of this project is to assess patient outcomes associated with the implementation of the Fitness, Lifestyle, and Optimal Wellness (FLOW) Program. The ultimate aim of this program is two-fold: increasing patient-reported wellness and improving objective health measurements. METHODS/STUDY POPULATION: The FLOW program consists of a multidisciplinary team of sports medicine physicians, nutritionists, fitness trainers, and clinical psychologists. Patients who choose to participate in the program undergo a comprehensive physician-guided assessment, including lifestyle and metabolic evaluation, biomarker profile, and body composition analysis. Based on the patient\u2019s goals and results of evaluation, he/she is then connected with other members of the FLOW team to develop a comprehensive plan and offer resources for potential improvements in physical activity, nutrition, and/or behavior. The patient will undergo follow-up assessments and questionnaires at three and six months to track their objective measurements and reported progress. RESULTS/ANTICIPATED RESULTS: The anticipated results of the FLOW program are an overall improvement in patient health and wellbeing. More specifically, we anticipate seeing increased levels of exercise from initial reported levels, as well as better nutrition habits. We expect to see improvements in follow-up body composition assessments, with gains in fat-free mass and decreased body fat, in addition to patient-reported improvements in behavioral health as measured by PHQ-9, GAD-2, and the Perceived Stress Scale. We will also assess reported sleep health with the hopes to see improvement in follow-up assessments. DISCUSSION/SIGNIFICANCE OF FINDINGS: The FLOW program is designed to address health inequities that disproportionately affect the Deep South. Through this program, we propose a new role of the primary care team in promoting healthy lifestyle habits and disease prevention through exercise, nutrition, and behavioral health services.Regulatory Science"} +{"text": "Increasingly, people have direct access to e-Health resources such as health information on the Internet, personal health portals, and wearable self-management applications, which have the potential to reinforce the simultaneously growing focus on self-management and wellbeing. To examine these relationships, we searched using keywords self-management, patient-targeting e-Health tools, and health as wellbeing. Direct access to the health information on the Internet or diagnostic apps on a smartphone can help people to self-manage health issues, but also leads to uncertainty, stress, and avoidance. Uncertainties relate to the quality of information and to use and misuse of information. Most self-management support programs focus on medical management. The relationship between self-management and wellbeing is not straightforward. While the influence of stress and negative social emotions on self-management is recognized as an important cause of the negative spiral, empirical research on this topic is limited to health literacy studies. Evidence on health apps showed positive effects on specific actions and symptoms and potential for increasing awareness and ownership by people. Effects on more complex behaviors such as participation cannot be established. This review discovers relatively unknown and understudied angles and perspectives about the relationship between e-Health, self-management, and wellbeing. Doctors have long been the gateway to diagnosis and health care, since they have privileged access to diagnostic tools and medical knowledge . IncreasThis direct access to e-Health resources is especially relevant for self-management for people with chronic diseases. Especially for people with chronic diseases, self-management has gained importance as an essential part of care and support, both within health service organizations, as well as in communities. It is considered a promising concept that, with right guidance and support, can empower people to address damaging health behaviors and address context challenges for better lifestyles . Indeed,In theory, patient-targeted e-Health tools can allow people to support self-management and reach their own health-related goals contributing to wellbeing. Yet the discourse and early evidence about patient targeted e-Health and its effect on self-management is mixed. Most scientific evidence pertains to intervention studies of health care provider-designed interventions supporting disease management and behavior change. A 2013 systematic review found the strongest evidence of positive effects for mobile health interventions for behavior change such as smoking cessation . A 2021 The explorative nature of the research question was best suited for a narrative review. A sampling of publications was done through keywords in three conceptual categories: (a) self-management; (b) patient-targeted e-Health tools; (c) health as wellbeing , and thrKeywords for category (a) included self-management; for (b) patient websites, wearables, digital self-management tools; for (c) health and wellbeing. Papers about the relationship between one concept and another or both of the other concepts were selected. Interventions that were designed by health care providers and embedded within a comprehensive care approach were not included. The inclusion criteria allowed assessment of all English published papers in the selected databases, including editorials, literature reviews, critical reflections, and intervention studies. There was no date restriction, because the concepts of self-management and health/wellbeing were well researched in the 1960s, while e-Health tools for patients were introduced with the development of smartphones in the new millennium. The selection of papers was based upon the information provided about the theoretical development and empirical evidence of one concept in relation with the other concept(s). Three databases were included: Web of Science was the most inclusive database for robust scientific papers from both the medical and social domain; medline for any additional references; Google Scholar for gray literature; and the Cochrane review database to collect synthesized evidence on these topics. Exclusion criteria were protocol papers, papers without full text available, papers not within the prime scope of interest.The combination of keywords resulted in 49 reviews in the Cochrane database, 661 publications in Web of Science, 158 in Google Scholar and 19 in Medline. A prisma chart visualizes the further selection process. Snowballing through reference lists and peer-suggestions enlarged the list . In the The pathways through which websites, wearables, and apps contribute to different aspects of self-management vary.Digitalization has allowed information and communication channels to be versatile, and new platforms and other creative digital outlets allow patients to acquire knowledge and skills in many different ways, suited to their personal profile, at their preferred time and place . This isOn the other hand, e-Health can also lead to uncertainty, stress, false reassurance, distrust, and avoidance, creating negative effects on self-management behavior. Uncertainties relate to the quality of information and to use and misuse of information. It is difficult to assess the relevance, quality, and reliability of data on the internet . Informasupport, which typically include information, educational, psychological, practical, and social support. Many digital and other programs focus on Lorig\u2019s set of self-management skills\u2014problem solving, decision making, resource utilization, forming a patient-health provider partnership and taking action [Self-management is mostly researched in the context of self-management g action . Reviewsg action and in pg action show thag action . Yet, thg action . ConceptThe influence of stress and negative social emotions on self-management is recognized by people themselves as a cauThe focus on self-management has developed concurrently with a broader vision of health, but the link between these concepts is complex. Ryan, Huta, and Deci formulated a theory on the relationship among motivation, self-regulation, and wellbeing , stating that eudaimonia\u2014or living well\u2014can be obtained by (a) pursuing goals that are intrinsically valued; and by (b) processes that are characterized by autonomy and awareness and thatPolicymakers, health care providers, and patients share the opinion that good self-management implies autonomy, pro-activeness, and responsibility and thatThe variety of e-Health tools and the diverse drivers for people to use them make it difficult to synthesize the between e-Health and wellbeing. For instance, people use smartwatches mostly for recreative purposes which might lead to satisfaction and potentially wellbeing. The addition of medical apps, such as heart rate monitors or anxiety reduction, has provided opportunities for people to gain insight into their condition and its mastering. People using specific apps report that their usage led to increased feelings of ownership .Health-related websites are also e-Health tools since they provide medical information and/or offer the option of social interaction on chat platforms. Experiences of people participating in interactions on such health platforms are generally positive . People However, e-Health can negatively affect wellbeing because of the potentially upsetting content, stress on how to use apps or navigate sites, and the fear to lose control over who has access to and sees the information. Web resources and search engines have increased in quality allowing more person-centered information , but theThrough a narrative review, this paper explores the complex relationships among patient-targeted e-Health tools, self-management, and wellbeing, discovering relatively unknown and understudied angles and perspectives. Most studies have examined and found proof of, the effects between patient-targeted e-Health and awareness and specific health\u2014behavior outcomes, being a part of self-management. The effects on behavior from a more complex perspective, for instance, participation and overall functioning, or emotion management, have not been established nor been unraveled. Observational studies and critical reflections point to the potential negative effects of expectations towards people to self-manage such as stress and shame, which in turn influence wellbeing.This paper has limitations. The data collection has been non-exhaustive. the analysis has not been structured. The exploratory research question about the nature of the relationships justified the approach of starting from broad keywords and snowballing to discover the domain. Not all available studies about self-management apps have been included, but the area has been generally covered. The review allowed us to observe an underrepresentation of studies about e-Health tools such as websites and wearables, and the paucity of evidence about the link with wellbeing. The selection of papers was guided by the research question, but the absence of strict inclusion criteria reduces the reproducibility of this process. The critical synthesis of the existing evidence is the result of a personal reflection, which is subject to selection and interpretation bias.Nevertheless, the review reveals important areas of research: impact of commercialized e-Health tools on wellbeing; the influence of emotions on self-management; determinants of self-management at individual and at collective levels, and its relationship with empowerment; and longitudinal studies on the influence of e-Health on complex behavior\u2019s. The effects of the digitalization of health care relationships have been examined ,71, but The review urges us to reflect upon the developments in the health and health care system. e-Health is changing health care systems and the relationship between patients and health care professionals , the orgPatient-targeted e-Health tools can allow people to support self-management and reach their own health-related goals contributing to wellbeing. However, this is hampered by a lack of clarity and empirical evidence about the positive and negative influences on these relationships. Our initial exploration of these relationships warrants further research especially about the (mitigation of) negative effects. Health care resources are available via multiplying channels to many actors, and people are using e-Health with or without professional support. This fundamentally changes the traditional primary care-based health system model. It re-emphasizes the need for a better understanding of the meaning of self-management and wellbeing. It urges researchers, policymakers and people to reflect how we\u2014at an individual and a societal level\u2014relate to e-Health and further digitalization."} +{"text": "Nature Communications 10.1038/s41467-021-23676-x, published online 7 June 2021.Correction to: The original version of this Article contained an error in the author affiliations.The affiliation of Jennifer Pett-Ridge with Life and Environmental Sciences Department, University of California Merced, Merced, CA was inadvertently omitted.This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "This dataset supports the findings of the vascular e-Learning during the COVID-19 pandemic survey (the EL-COVID survey). The General Data Protection Regulation (GDPR) of the European Union was taken into consideration in all steps of data handling. The survey was approved by the institutional ethics committee of the Primary Investigator and an online English survey consisting of 18 questions was developed ad-hoc. A bilingual English-Mandarin version of the questionnaire was developed according to the instructions of the Chinese Medical Association in order to be used in mainland People's Republic of China. Differences between the two questionnaires were minor and did affect the process of data collection. Both questionnaires were hosted online.The EL-COVID survey was advertised through major social media. All national and regional contributors contacted their respective colleagues through direct messaging on social media or by email. Eight national societies or groups supported the dissemination of the EL-COVID survey.The data provided demographics information of the EL-COVID participants and an insight on the level of difficulty in accessing or citing previously attended online activities and whether participants were keen on citing these activities in their Curricula Vitae. A categorization of additional comments made by the participants are also based on the data.The survey responses were filtered, anonymized and submitted to descriptive analysis of percentage. Researchers from other relevant medical fields could also use these data to support any necessary transition from traditional education and training to e-Learning.\u2022Our data could be compared to or be complementary to similar data from newer studies.\u2022These data may assist medical schools or institutions to implement the necessary changes for a partial or full transition to e-Learning.1The dataset provides useful information based on survey data of the EL-COVID regarding the vascular e-Learning appreciation, advantages and disadvantages during the pandemic. The demographics of the 856 participants are presented in The difficulty of accessing previously attended online training or educational activities varies from very easy to impossible. This process was considered very easy or easy in 53% and 16%, respectively. On the contrary, 7% and 9% of participants considered it very hard or impossible, respectively .Fig. 1DeThe difficulty of citing a previously attended online activity varies as well. Most of the participants (40%) would not cite an online activity . The resAccording to the responses to our survey, 19% of participants would list all of the online activities they have attended in their Curricula Vitae, while 24% would not list any of them. The majority (53%) would list only selected activities .Fig. 3LiApart from the questions of the survey, there were 328 free comments on vascular e-Learning during the pandemic. These comments were classified according to their subject and they are listed in 2This international survey was performed using a cross-sectional method to determine the appreciation, advantages and disadvantages of vascular e-Learning during the COVID-19 pandemic.The official language of the survey was English. The General Data Protection Regulation (GDPR) of the European Union was taken into consideration in preparing all materials of this research. The survey was approved by the ethics committee of the Primary Investigator's institution.In our study, a vascular e-Learning activity or distance learning activity was defined as any educational or training activity that is performed exclusively online (either synchronous or asynchronous) and the main topic falls under the scope of vascular or endovascular disciplines.The pandemic period was defined as the period between March 15th and May 15th, 2020 for Europe. For other regions, this period may slightly vary according to imposed measures by each government and epidemiological data from each region. This statement was posted in the official research page and in the two survey forms.The design of our study required only vascular surgeons and vascular trainees to complete the survey, something that was a challenge on its own account. There are different definitions on who is a vascular surgeon or a vascular trainee. In some regions, a vascular surgeon would perform an open procedure but not an endovascular one. In other parts of the world, general surgeons with vascular training would perform some procedures, but not others. In some countries, trainees often have to go through a core surgical training program before they go ahead with vascular training, but they are not considered vascular trainees until they do the latter. So, the EL-COVID team decided not to strictly define who is a vascular surgeon, but rather used a vague definition of any surgeon who performs the majority of open and endovascular procedures. In a similar manner, a vascular trainee is any physician enrolled at any stage of an official vascular surgery training program.An online questionnaire consisting of 18 questions was developed ad-hoc; three questions were on demographics, 14 on the e-learning experience and opinion, and one field for the participants\u2019 email address. Filling-in the email was voluntary aiming in assisting in the verification and validation of our results. Validation of the data was initially performed by ignoring suspicious results recorded from the same computer within a small period of time and individuals who had participated more than once . All data that could lead to identifying specific individuals who participated in the survey were not shared with any parties and were deleted once data validation was complete. A single copy containing all raw data is saved by the GDPR data protection officer (NP) for future reference, if questions of data validity were risen. A bilingual English-Mandarin version of the same questionnaire was created according to the instructions of the Chinese Medical Association in order to be used in mainland People's Republic of China (PR China). Apart from the Chinese text, the only difference between the original and the bilingual questionnaires was that in the latter there was no country of practice field, as it was developed only for use within mainland PR China. Both questionnaires are available as supplementary material.The original questionnaire was hosted on Google Forms , while the bilingual version was hosted on SurveyLab .The online questionnaires were available on both hosts for a period of four months. The EL-COVID survey was advertised through social media (so.me.); primarily in LinkedIn and secondarily in Twitter and Facebook . All national and regional contributors were asked to contact their respective colleagues through direct messaging on any so.me. platform or by email. Thirty-eight international and national societies were contacted in order to support the EL-COVID research by sending their members a link to the survey; with eight of them supporting our research and two declining. The above information was described in more detail in the official EL-COVID webpage hosted at med-pie.com on a non-commercial basis. All data analysis was performed using Microsoft Excel for Mac version 16.16.27.This study has a number of limitations. Since the questionnaires were only hosted online and the survey information was disseminated through so.me., data collection might be biased against vascular surgeons and trainees who are not active in so.me. or familiar with online questionnaires. The second limitation is that it is impossible to know the exact number of vascular surgeons and trainees, since there are different vascular training curricula around the globe and vascular surgery is not an independent medical specialty in some countries. Therefore, the number of vascular professionals worldwide can only be estimated with approximation. Our sample is the largest one compared to all other relevant studies, but the degree it represents the global vascular society is only an estimation.http://dx.doi.org/10.17632/fv5ztss3yf.2.The survey was approved by the ethics committee of the institution of the primary investigator. The European Union's GDPR was taken into consideration in preparing all materials of this research. No funding was received. Anonymized data are archived both offline and online at N. Patelis: Concept, design, data collection, data analysis, writing, critical appraisal of the manuscript; T. Bisdas: Design, data analysis, critical appraisal of the manuscript; Z. Jing: Data collection, data analysis, critical appraisal of the manuscript; J. Feng: Data collection, data analysis, critical appraisal of the manuscript; M. Trenner: Data collection, data analysis, writing, critical appraisal of the manuscript; N. Nugroho: Data collection, data analysis, critical appraisal of the manuscript; P.E. Reis: Data collection, data analysis, critical appraisal of the manuscript; S. Elkouri: Data collection, data analysis, critical appraisal of the manuscript; A. Lecis: Data collection, data analysis, critical appraisal of the manuscript; L. Karam: Data collection, data analysis, critical appraisal of the manuscript; D. Le Roux: Data collection, data analysis, critical appraisal of the manuscript; M. Ionac: Data collection, data analysis, critical appraisal of the manuscript; M. Berczeli: Data collection; V. Jongkind: Data analysis, writing, critical appraisal of the manuscript; K.K. Yeung: Design, data collection, critical appraisal of the manuscript; A. Katsargyris: Data collection, critical appraisal of the manuscript; E. Avgerinos: Data collection, writing, critical appraisal of the manuscript; D. Moris: Data collection, writing, critical appraisal of the manuscript; A. Choong: Data collection, data analysis, critical appraisal of the manuscript; J.J. Ng: Data collection, data analysis, critical appraisal of the manuscript; I. Cvjetko: Data collection, critical appraisal of the manuscript; G. Antoniou: Data collection, writing, critical appraisal of the manuscript; P. Ghibu: Data collection, critical appraisal of the manuscript; A. Svetlikov: Data collection, writing, critical appraisal of the manuscript; F. Gallardo Pedrajas: Data collection, critical appraisal of the manuscript; H. Ebben: Data collection, critical appraisal of the manuscript; H. Stepak: Data collection, critical appraisal of the manuscript; A. Chornuy: Data collection, critical appraisal of the manuscript; S. Kostiv: Data collection, critical appraisal of the manuscript; S. Ancetti: Data collection, critical appraisal of the manuscript; N. Tadayon: Data collection, critical appraisal of the manuscript; A. Mekkar: Data collection, critical appraisal of the manuscript; L. Magnitskiy: Data collection, critical appraisal of the manuscript; L. Fidalgo-Domingos: Data collection, critical appraisal of the manuscript; S. Matheiken: Data collection, critical appraisal of the manuscript; E. Sarutte Rosello: Data collection, critical appraisal of the manuscript; A. Isik: Data collection, critical appraisal of the manuscript; G. Kirkilesis: Data collection, critical appraisal of the manuscript; K. Kakavia: Data collection, critical appraisal of the manuscript; S. Georgopoulos: Data analysis, critical appraisal of the manuscript.Nikolaos Patelis and Sean Matheiken are co-founders of the Med-PIE group (med-pie.com)Otherwise, the authors declare that they have no known competing financial interests or personal relationships which have, or could be perceived to have, influenced the work reported in this article.No funding was received."} +{"text": "Major depressive disorder (MDD) and anxiety disorders are common and result in considerable suffering and economic loss. People suffering from major depressive disorder and/or anxiety disorders are commonly encountered in the primary care setting. Unfortunately, most people with these disorders remain either untreated or inadequately treated; current data suggest that general practitioners fail to diagnose up to half of cases of major depressive disorder or anxiety. There is a need for screening tools that will help physicians and other professionals in primary care recognize and adequately treat major depressive disorder and anxiety disorders. While the currently-available self-report screening instruments have been demonstrated to be reliable and valid, there remain considerable barriers to their widespread use in primary care.The purpose of the present study is to report preliminary validation data for a freely-available, brief, Web-based, self-report screener for major depressive disorder and anxiety disorders.The Web-Based Depression and Anxiety Test (WB-DAT) was administered to 193 subjects who presented for assessment and/or treatment in ongoing research projects being conducted at the Mood and Anxiety Program and Clinical Research Department at the Centre for Addiction and Mental Health in Toronto, Ontario, Canada. Subjects completed the Web-based screening instrument and were subsequently interviewed with the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV) Axis I Disorders (SCID-I/P). The diagnostic data from the screening instrument were then compared with the data from the individual's SCID-I/P interview. Diagnostic concordance between SCID-I/P diagnoses and the Web-Based Depression and Anxiety Test were assessed using Cohen's kappa, sensitivity, specificity, positive predictive value, negative predictive value, and efficiency.Agreement ranged from acceptable to good (0.57-0.70) for major depressive disorder, panic disorder with and without agoraphobia (PD+/-AG), social phobia/social anxiety disorder, obsessive compulsive disorder (OCD), generalized anxiety disorder (GAD), and post traumatic stress disorder (PTSD). With the exception of generalized anxiety disorder, the sensitivity (0.71-0.95) and specificity (0.87-0.97) for the major diagnostic categories assessed by the Web-Based Depression and Anxiety Test were good. The sensitivity for generalized anxiety disorder was somewhat lower (0.63) but acceptable. Positive predictive values were good (0.60-0.75) for major depressive disorder, obsessive compulsive disorder, generalized anxiety disorder, and post traumatic stress disorder, and acceptable for panic disorder with and without agoraphobia and for social phobia/social anxiety disorder.These preliminary data suggest that the Web-Based Depression and Anxiety Test is reliable for identifying patients with and without major depressive disorder and the anxiety disorders of panic disorder with and without agoraphobia, social phobia/social anxiety disorder, obsessive compulsive disorder, and post traumatic stress disorder. Further research is required in a larger sample in primary care. Major depressive disorder (MDD) and the anxiety disorders are common and result in significant suffering, lost opportunity, and economic loss. With a prevalence rate of approximately 5% worldwide, MDD is the most common mood disorder . EstimatThe Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV) recognizes a number of distinct anxiety disorders, including specific phobias, social phobia/social anxiety disorder (SP), panic disorder (PD) with and without agoraphobia (PD+/-AG), AG without a history of panic, generalized anxiety disorder (GAD), obsessive-compulsive disorder (OCD), and post-traumatic stress disorder (PTSD). Anxiety disorders are among the most-prevalent psychiatric illnesses. According to the National Comorbidity Survey the lifetime prevalence for all categories of anxiety disorders in the United States is 24.9% . AlthougAnxiety disorders have high rates of comorbidity with other psychiatric disorders including other anxiety disorders, MDD, and substance abuse/dependence. Anxiety disorders often occur with MDD. For example, MDD occurs in up to 60% of people with anxiety disorders . ComorbiIn North America, primary care/family medicine practitioners are the primary providers of first-line treatment for MDD and anxiety disorders . People Current data suggest that general practitioners fail to diagnose up to half of cases of depression or anxiety . This siThere are barriers to better assessment and treatment of MDD and the anxiety disorders in primary care, including a lack of recognition and adequate treatment in primary care, such as a lack of brief, sensitive, easy-to-administer, and easy-to-interpret self-report psychiatric-screening instruments. Without adequate detection and an accurate diagnosis, there cannot be adequate treatment. Establishing an accurate primary diagnosis is important in guiding the specific method and course of treatment . CurrentIn psychiatry, structured diagnostic interviews are the standard for diagnostic accuracy and are widely employed in research settings. Structured interviews such as the Structured Clinical Interview for DSM-IV Axis I Disorders (Version 2.0/Patient Form) (SCID-I/P) and the In response to increasing demands for diagnostic precision and accountability in nonresearch clinical settings, there are now reliable and valid screening instruments available for use in primary care including the Primary Care Evaluation of Mental Disorders (PRIME-MD) , SymptomWhile the currently-available self-report screening instruments have been demonstrated to be reliable and valid, there remain considerable barriers to their widespread use in primary care. First, many of the available instruments are narrow in their scope of assessment. For example, there are a large number of 1-page screening instruments designed to assess for the symptoms of MDD, PD, PD+/-AG, social anxiety disorder/social phobia, OCD, GAD, or PTSD. However, given the high rates of comorbidity among these disorders, instruments that assess for only 1 of them are of dubious utility. A major problem with all of the better and broadly-focused DSM-IV screening tools is that they are not freely available. In addition, these instruments all require laborious scoring and interpretation by a health care professional. Given these serious barriers to ease of use, they are unlikely to be widely adopted in primary care no matter how good they are.The Internet provides an excellent medium for providing patients and health care professionals in primary care access to a brief, algorithm-scored, easily-interpretable self-report screening test for MDD and the anxiety disorders. There are a large number of self-report screeners for anxiety and depression available on the Internet. Unfortunately, they are all subject to the same limitations as the currently-available paper-and-pencil tests insofar as they are all either too limited in scope, not easily scored or interpreted, and/or not freely available. None provide both a broad screen of MDD and the anxiety disorders and few provide any kind of print function that might facilitate a discussion of symptoms with a health care professional in primary care.http://www.depressioncenter.net/depressiontest) [http://www.paniccenter.net/anxietytest) [Van Mierlo Communications Consulting Inc has recently designed a screening test for MDD and the anxiety disorders that is freely available on the Internet. The screener is currently available as The Depression Test at The Depression Center (iontest) , and slietytest) .This test, the Web-Based Depression and Anxiety Test (WB-DAT) was designed to be a brief, freely-available, Web-based, self-report screening tool for MDD and the anxiety disorders compatible with the DSM-IV and The International Classification of Diseases and Related Health Problems, tenth revision (ICD-10) diagnostic systems . As a scBased on their responses to 11 broad preliminary questions based on DSM-IV criteria central to the diagnoses of MDD and each of the anxiety disorders, users are presented with several additional questions for each disorder based on DSM-IV criteria. The result is an algorithm-generated personalized \"final report,\" which summarizes the individual's responses relating to the major diagnostic categories. The final report was designed to be printed and shared with a health care professional.The WB-DAT was designed to provide a summary of standard diagnostic information in order to initiate and encourage a discussion of specific anxiety and depression symptoms between patients and health care professionals. As a result, there are few diagnostic algorithms to limit the number of diagnoses a health care professional might query. Thus, for example, if a patient meets screening criteria for MDD, GAD, and OCD, the screener summary will report symptoms of MDD, GAD, and OCD, leaving the diagnostic decision regarding the primary diagnosis and focus of treatment to the health care professional.In deciding what disorders to screen for in primary care, developers of the test were guided by the diagnostic criteria described in DSM-IV and ICD-10. As a result, the WB-DAT includes screening modules for MDD, PD+/-AG, AG without a history of panic, OCD, social phobia/social anxiety disorder, GAD, PTSD, and acute stress disorder (ASD). The focus of the WB-DAT is on current, rather than past (or lifetime), symptoms and distress/impairment.Although the WB-DAT has considerable face validity, it is important that the instrument's operating characteristics be evaluated by determining the agreement between the WB-DAT screener diagnoses and diagnoses as made by SCID-I/P. Thus, the purpose of the present study is to report on the operating characteristics of the WB-DAT as compared with gold-standard diagnoses obtained by the SCID-I/P. The WB-DAT was also designed to include additional screening modules for agoraphobia without a history of panic, acute stress disorder, specific phobia, and a number of subsyndromal symptom profiles that may aid health care professionals in primary care to reach diagnostic conclusions. However, due to the relatively-small sample size in this study we report here only data for the major diagnostic categories .The WB-DAT was administered to 193 subjects. All subjects were 18 years of age or older. The sample consisted of 79 (40.9%) men and 114 (59.1%) women. On average, subjects were 40.92 (SD = 12.61) years of age. Subjects with dementia, mental retardation, or serious medical illnesses were excluded.Subjects were recruited from individuals who presented for assessment and/or treatment in ongoing research projects being conducted at the Mood and Anxiety Program and Clinical Research Department at the Centre for Addiction and Mental Health (CAMH) in Toronto, Ontario, Canada. Projects included 2 ongoing studies of the treatment of MDD, and a study of DSM-IV symptoms and personality in social and problem gamblers. In addition to the standard assessments conducted in the study, interested subjects were asked to consent to participate in the validation study of the WB-DAT.Subjects completed the WB-DAT using a pseudonym and were subsequently interviewed with the SCID-I/P. The diagnostic data from the WB-DAT were then compared with the data from the individual's SCID-I/P interview. The SCID-I/P was administered by MA-level and PhD-level psychology graduate students who had received formal standardized training, including observing expert-conducted interviews and being observed conducting interviews. Such training has been reported to produce high diagnostic agreement for the DSM-IV Axis I disorders . This stDiagnostic concordance with the SCID-I/P was assessed for each Axis-I disorder assessed by the WB-DAT using Cohen's kappa, sensitivity, specificity, positive predictive value, negative predictive value, and efficiency - 34. CohSubjects received an average of 0.99 (SD = 1.45) diagnoses according to the WB-DAT and 0.79 (SD = 1.17) diagnoses according to the SCID-I/P. However, only 79/193 (40.9%) of the sample met WB-DAT criteria for 1 or more disorders, and only 78/193 (40.4%) met SCID-I/P criteria for 1 or more disorders. The base rates for both acute stress disorder and AG without a history of panic were too low to permit evaluation of the performance of the WB-DAT for these disorders. The prevalence rates for MDD, any anxiety disorder, and any disorder according to the WB-DAT and the SCID-I/P for the sample are shown in The measures of agreement for the WB-DAT as compared with the SCID-I/P criterion for the DSM-IV Axis I disorders assessed are shown in These are preliminary data from a sample of subjects drawn from 2 studies of the treatment of MDD and from a community study of social and problem gamblers. Thus, the results of this study should be interpreted with some caution. However, these preliminary data suggest that the WB-DAT was reasonably accurate in identifying patients who met SCID-I/P criteria for MDD, SP, OCD, and PTSD. The WB-DAT was somewhat less accurate in identifying subjects with GAD, although this is likely due to the small sample size and the considerable comorbidity between MDD and GAD, as 35.41% of subjects who met SCID-I/P criteria for MDD also met SCID-I/P criteria for GAD.Given the relatively small sample size in this study it is important to note that the Cohen's kappa, sensitivity, and specificity measures for the diagnoses of \"any anxiety disorder\" and \"any disorder\" were excellent. Thus, given that the true purpose of the WB-DAT is to produce output that can help initiate and encourage a discussion of symptoms and concerns between patients and health care providers in primary care, it appears to have the potential to be a useful tool in primary care.In summary, the WB-DAT appears to do a reasonably good job of identifying people with MDD and/or an anxiety disorder. However, the results of this study require support from a larger validation study in primary care. The use of Web-based technology allows for constant improvements in screening modules and diagnostic algorithms in response to feedback from the results of validation studies. With the potential for continued development and validation, the WB-DAT provides a unique opportunity to make an important contribution to increasing recognition of MDD and the anxiety disorders in primary care."} +{"text": "Chronic in vivo imaging of fluorescent-labeled neurons in adult mice reveals extension and retraction of dendrites in GABAergic non-pyramidal interneurons of the cerebral cortex. PLoS Biology, volume 4, issue 2, DOI:10.1371/journal.pbio.0040029InThe authors wish to add to the Acknowledgments that Emery N. Brown's work on the project was supported in part by a grant from the National Institute on Drug Abuse of the National Institutes of Health."} +{"text": "To remain competitive, the providers of medical and health-related information must continually adapt their Web sites to new market demands and trends. Successful adaptation depends, among other things, on understanding users' needs. The Health on the Net Foundation (HON) has been conducting regular surveys of user-traffic since 1997. The fourth and latest in the series, conducted through the months of March and April, 1999, obtained 4,437 responses, compared to 1,863 responses obtained by the third survey in May and June, 1998. Thanks to these surveys, a broad user profile for Web-based medical and health-related information is emerging. The electronic questionnaire has remained largely the same since HON first posted it on its Web site in February-March, 1997. As in the past, results of the current survey are intended for use by all interested organisations. This article examines some major trends in user responses since then.The survey uses non-probabilistic sampling, and cannot ensure that participants are representative of the total medical and health information user community on the Web. Additional links to the HON survey posted by \"friendly\" Web sites and others helped boost participation. The questionnaire is designed for completion within three to four minutes. It starts with some basic user-related questions . Next come a number of statements and a multiple choice of relevant answers, from \"Strongly agree\" to \"Strongly disagree\", for simple clicking with the mouse. HON added two new questions to the March-April, 1999 questionnaire: \"What type of site do you first go to?\", offering six multiple-choice options, and \"What are your three most preferred Web sites?\". The latter is the only open question in the survey. It requires users to provide their three \"favourite\" URLs. Respondents are also given an opportunity to leave spontaneous messages: in the last survey, 782 left a wide variety of remarks.Results and Discussion:The survey suggests an important growth in Internet use for medical and health-related information by users in Europe. Only 18% cannot find information in their primary language. The May-June, 1998 HON survey showed 71% of respondents were North American and only 18% European, while the March-April, 1999, survey showed 48% North American users and 28% Europeans. Further, most European users (79%) were medical professionals and male (67%), an important difference to North American users .The survey suggests that, generally speaking, users are mature to middle-aged, relatively content with the variety of medical and health-related information on the Internet, increasingly multi-national in terms of country of origin, and increasingly concerned about quality."} +{"text": "The objective of this study was to evaluate the efficacy of atomoxetine, a new and highly selective inhibitor of the norepinephrine transporter, in reducing symptoms of attention-deficit/hyperactivity disorder (ADHD) among adults by using drug-placebo response curve methods.N = 280; Study II, N = 256) with DSM-IV-defined ADHD who were recruited by referral and advertising. Subjects were randomized to 10 weeks of treatment with atomoxetine or placebo, and were assessed with the Conners Adult ADHD Rating Scales and the Clinical Global Impression of ADHD Severity scale before and after treatment.We analyzed data from two double-blind, placebo-controlled, parallel design studies of adult patients (Study I, Those treated with atomoxetine were more likely to show a reduction in ADHD symptoms than those receiving placebo. Across all measures, the likelihood that an atomoxetine-treated subject improved to a greater extent than a placebo-treated subject was approximately 0.60. Furthermore, atomoxetine prevented worsening of most symptom classes.From these findings, we conclude that atomoxetine is an effective treatment for ADHD among adults when evaluated using several criteria. Several compounds are now recognized as effective treatments for the major symptoms of attention-deficit/hyperactivity disorder (ADHD) in adulthood. The most effective of these include methylphenidate and dextroamphetamine (or mixed dextro- and levoamphetamine); however, the use of other agents, such as bupropion and desipramine, has also received some support. In addition to these, atomoxetine, a highly selective noradrenergic reuptake inhibitor with little affinity for other neurotransmitter systems , has beeet al. point to the [The drug-placebo response curve is constructed in the following six steps: 1.) Choose an outcome variable, for example the change in CAARS Inattention score from baseline to the end of the study; 2.) At each observed score, calculate separately for the drug and placebo groups the proportion of subjects having that score or a better score. For CAARS change scores, therapeutic change is indicated by negative numbers, i.e., the probability that drug will outperform placebo.The drug-placebo response curve is a graphical method of describing results from a clinical trial, not a statistical test. It is most sensibly used to describe an effect that has been demonstrated with appropriate statistical tools. Nevertheless, the drug-placebo response curve's roots in (ROC) analysis motivate the computation of one statistic, the AUC, which is computed through integration. The area under the drug-placebo response curve ranges from 0.5 to 1.0 (when the drug is completely effective and the placebo has no effect). The AUC is a useful index of clinical significance because it equals the probability that a randomly selected member of the drug group will have a better result than a randomly selected member of the placebo group ,13, i.e.e.g., a clinically meaningful cut-point) can be determined as the proportion of drug-responders to placebo-responders at any given cut-point. Fourth, the ability of a drug to improve functioning vs. prevent worsening of functioning can be determined as the proportion of drug-responders to placebo-responders at the outcome score representing no change.In summary, the placebo-response curve provides four pieces of clinically relevant data not typically available from traditional statistical analyses of outcomes data. First, the effect size of a drug on an outcome measure can be determined as the distance between the curve and the line of no effect at any given cut-point. Second, the ratio of drug responders to placebo responders across the range of outcomes can be determined as the area under the curve. Third, the likelihood of a drug to elicit a specific outcome (Stephen V. Faraone, PhD.- Stephen Faraone receives research funding from Lilly, McNeil and Shire.Joseph Biederman, MD.- Joseph Biederman receives research support from the following sources: Shire Laboratories, Inc and Eli Lilly & Company, Pfizer Pharmaceutical, Cephalon Pharmaceutical,, Janssen Pharaceutical, Neurosearch. Pharmaceuticals, Stanley Medical Institute, Lilly Foundation, Prechter Foundation, NIMH, NICHD and NIDADr. Joseph Biederman is a speaker for the following speaker's bureaus: Eli Lilly & Company, Pfizer Pharmaceutical, Novartis Pharmaceutical, Wyeth Ayerst, Shire Laboratories Inc, McNeil Pharmaceutical, and Cephalon PharmaceuticalDr. Joseph Biederman is on the advisory board for the following pharmaceutical companies: Eli Lilly & Company, CellTech, Shire Laboratories Inc, Novartis Pharmaceutical, Noven Pharmaceutical, McNeil Pharmaceuticals, Janssen, Johnson & Johnson, Pfizer, and Cephalon PharmaceuticalsThomas Spencer, MD- Dr. Thomas Spencer receives research support from the following sources: Shire Laboratories, Inc and Eli Lilly & Company, Glaxo-Smith Kline, Pfizer Pharmaceutical, McNeil Pharmaceutical, Novartis Pharmaceutical, and NIMHDr. Thomas Spencer is a speaker for the following speaker's bureaus: Glaxo-Smith Kline, Eli Lilly & Company, Novartis Pharmaceutical, Wyeth Ayerst, Shire Laboratories Inc, McNeil PharmaceuticalThomas Spencer is on the advisory board for the following pharmaceutical companies: Shire Laboratories, Inc and Eli Lilly & Company, Glaxo-Smith Kline, Pfizer Pharmaceutical, McNeil Pharmaceutical, and Novartis PharmaceuticalDr. David Michelson, MD- David Michelson is a Lilly employeeLenard Adler, MD- Lenard Adler receives grant and Research Support, is a Consultant or on Advisory Boards: Abbott Laboratories, Bristol-Myers Squibb, Eli Lilly and Co., McNeil/Johnson & Johnson, Merck & Co., Inc., Neurosearch, Novartis Pharmaceuticals Corp., Pfizer Labs, Cortex Pharmaceuticals, Cephalon and Shire PharmaceuticalsFred Reimherr, MD-Fred Reimherr has been part of Lilly advisory board.Stephen J Glatt, PhD- Stephen Glatt has no conflicts of interest to declare.Stephen V. Faraone, PhD- Steve Faraone contributed to the analysis and interpretation of the data, the drafting and revision of the manuscript.Joseph Biederman, MD- Joseph Biederman contributed to the analysis and interpretation of the data, the drafting and revision of the manuscript.Thomas Spencer, MD- Thomas Spencer contributed to the conception and design of the study, the acquisition of data, interpretation of data and drafting/reviewing the manuscript.Lenard Adler, MD- Lenard Adler contributed to the conception and design of the study, the acquisition of data, interpretation of data and drafting/reviewing the manuscript.David Michelson, MD- David Michelson contributed to the study conception design, the data acquisition as well as critical reviewing of the manuscript.Fred Reimherr, MD- Fred Reimherr contributed to the study conception design, the data acquisition as well as critical reviewing of the manuscript.Stephen J Glatt, PhD- Stephen Glatt contributed to the analysis and interpretation of the data, and the drafting and revision of the manuscript."} +{"text": "The objective of this program announcement (PA) is to encourage research that bridges the areas of inorganic chemistry and medicine in continuation of PA-01-071. The National Institute of General Medical Science (NIGMS) is joined in this announcement by the NIEHS and the NIH Office of Dietary Supplements (ODS).The mechanisms by which organisms control transition metal ions and the roles of these metals in cellular regulation and signaling in health and disease are of principal interest. The interactions of synthetic inorganic complexes with living systems and their components are an additional area of interest. These areas are linked by the need to involve researchers having a deep understanding of inorganic chemistry in medically relevant research. Much of the work is expected to involve collaborations including chemists, biologists, and medical researchers. The results will be relevant to understanding the mechanisms of metal handling by biological systems and the basic cellular roles underlying the nutritional requirement for essential metals. It is expected that this research will also contribute to the identification of new targets for drug discovery, diagnostics, and future therapeutic approaches involving metal complexes, although drug development, per se, is not a focus of the program.A higher-order problem presents itself in understanding how the genome-encoded components and the other molecules are constituted in networks of interacting molecules with particular distributions in time and space. Advances in imaging techniques and analytic methods are beginning to yield copious quantitative and spatial data on specific molecules in biological systems. Knowledge of the network and changes in its components over time, and the local rules by which the individual components distribute material and information, will substantially advance our knowledge.Studies of metalloenzyme structure and function, mechanisms of action, and inhibition are currently well supported and produce results that are utilized in the design of new diagnostic and therapeutic products. Additional stimulation of this area is not needed. In contrast, work in other areas of bioinorganic chemistry lags behind its potential application to human health. These areas include 1) mechanisms of metal metabolism as well as the roles of metals in regulation of cell function and cell\u2013cell interaction, and 2) basic research toward diagnostic and therapeutic applications of metal complexes and of metal chelators and toward exploiting the unique properties of metals for therapeutic applications. The emphasis of this announcement is on the ions, complexes, and organometallic compounds of the transition metals known as lanthanides and actinides, post-transition metals, and metalloid elements.Metal metabolism is emerging as an exciting area of cell biology and a potential area for therapeutic intervention. Normal metal metabolism appears to maintain free metal ion concentrations at a very low level and to deliver metals very selectively to their sites of action, while maintaining tight control over their reactivity. Aberrant metal metabolism contributes to pathological conditions such as Menkes\u2019 disease, Wilson\u2019s disease, and hemochromatosis. Intercepting normal metalation reactions may be a way to control metalloprotein activity. Metals may also be associated with the pathology of protein aggregates such as those formed by prions and in Alzheimer\u2019s disease. Metals have also emerged as important sensors and transducers of information with roles in regulation and neurotransmission.Areas of interest include 1) improved metal ion sensors to study cellular metal ion concentrations and localization; 2) reagents suitable to manipulate those concentrations; 3) identification and characterization of the macromolecular players and vesicular compartments involved in metal ion homeostasis and metal trafficking; 4) elucidation of the roles of metals in cell regulation, signal transduction, and cell\u2013cell signaling; 5) identification and understanding of mRNAs and metal-, oxygen-, and redox-responsive transcriptional and translational regulators, and their potential as therapeutic targets; 6) elucidation of the mechanistic roles of essential trace elements for which metabolic functions are not yet clearly established; 7) analytical tools that accurately monitor biologically important pools, storage pools, and the chemical speciation of metals; 8) biomarkers of exposure and mechanisms of metal toxicity; 9) biomarkers for variable susceptibility to metal toxicity in the human population; and 10) chelation chemistry that can serve as the foundation for therapies to ameliorate aberrant metal accumulations and the effects of toxic exposures.The therapeutic application of metal complexes is an underdeveloped area of research. Basic principles to guide the development of metallopharmaceuticals are lacking. Metal-containing agents may offer unique therapeutic opportunities. However, significant obstacles, including potential metal accumulations and toxicities, require further research before the promise of medicinal inorganic chemistry can be realized.Metal complexes may be useful as research probes of biological function, as intermediary lead compounds in the development of non\u2013metal-containing therapeutics, and as potential diagnostic and therapeutic agents. Opportunities exist to exploit the unique properties of metal complexes, to measure and/or alter cellular functions. The actions of these compounds may provide insights that are different from those that can be achieved through other chemical, biochemical, or genetic manipulations. Similarly, the actions of metal complexes in whole living organisms are expected to differ in general from the actions of non\u2013metal-containing agents and may offer unique research, diagnostic, or therapeutic opportunities. Principles are needed for the design of safe metal-containing therapeutics.Another goal of this program is to utilize the power of inorganic chemistry to provide new knowledge of and new approaches for intervention in biological systems. Still another goal is to improve understanding of the reactions of metal complexes in living systems to improve the specificity of these interactions and gain control over the potential toxicity of synthetic metal complexes. The long-term goal is to establish the basic principles of an inorganic medicinal chemistry that will allow for rational design and screening of potential metallopharmaceuticals in the future.in vivo; 3) uptake of metal complexes into cells and delivery to specific cellular compartments; 4) interactions of metal complexes with specific enzymes and receptors; 5) mechanisms by which synthetic metal complexes recruit cell cycle, signal transduction, and other metabolic pathways to alter cell functions; and 6) structure\u2013activity relationships for ligand design to control metal complex activity and stability in vitro and in vivo.Areas of interest include 1) reactions of metal complexes with cellular constituents ; 2) reactions of metal complexes within the cellular milieu and The NIH Metals in Medicine meeting report includes a list of specific research opportunities and challenges. This list is intended to be illustrative, not exhaustive. Investigator-initiated ideas are welcome on any subject that will contribute to the objectives listed in this PA.Research encouraged by this announcement may utilize any appropriate experimental organisms or model systems. For some problems, interesting discoveries may be found in microorganisms from unusual environments and atypical experimental organisms. For other problems, yeast, common invertebrate and vertebrate model organisms, and human cell/tissue cultures may be appropriate. Investigators considering human clinical trials are strongly encouraged to contact the program staff.http://grants.nih.gov/grants/funding/r21.htm. For descriptions of the P01 grant mechanism see http://www.nigms.nih.gov/funding/grntmech.html#b (NIGMS) and http://www.niehs.nih.gov/dert/programs/p01.htm (NIEHS).This funding opportunity will use the regular research (R01), exploratory research (R21), and program project (P01) award mechanisms. For a description of the R21 grant mechanism see http://grants.nih.gov/grants/funding/modular/modular.htm). Specifically, if you are submitting an application with direct costs in each year of $250,000 or less, use the modular budget format described in the PHS 398 application instructions, available at http://grants1.nih.gov/grants/funding/phs398/phs398.html in an interactive format. Otherwise, follow the instructions for nonmodular research grant applications. For further assistance, contact GrantsInfo at 301-435-0714 (telecommunications for the hearing impaired: TTY 301-451-0088) or by e-mail: GrantsInfo@nih.gov.This funding opportunity uses just-in-time concepts. It also uses the modular as well as the nonmodular budget formats (see http://www.dnb.com/us/. The D&B number should be entered on line 11 of the face page of the PHS 398 form. Applications must be submitted on or before the receipt date described at http://grants.nih.gov/grants/funding/submissionschedule.htm. The complete version of this PA is available at http://grants.nih.gov/grants/guide/pa-files/PA-05-001.html.Applications must be prepared using the PHS 398 application instructions and forms (rev. 5/2001). Applications must have a Dun & Bradstreet (D&B) Data Universal Numbering System number as the universal identifier when applying for federal grants or cooperative agreements. This number can be obtained by calling 1-866-705-5711 or online at Contact: Peter C. Preusch, Division of Pharmacology, Physiology, and Biological Chemistry, NIGMS, Bldg 45, Rm 2AS.43C, MSC 6200, Bethesda, MD 20892-6200 USA, 301-594-5938, fax: 301-480-2802, e-mail: preuschp@nigms.nih.gov; Claudia Thompson, NIEHS, DERT/OPD/CEMBB, MD EC-21, PO Box 12233, Research Triangle Park, NC 27709 USA, 919-541-4638, fax: 919-541-4606, e-mail: thompso1@niehs.nih.gov; Becky Costello, ODS, NIH, Bldg 31, Rm 1B25, MSC 2086, Bethesda, MD 20892-2086 USA, 301-435-2920, fax: 301-480-1845, e-mail: costellb@od.nih.gov. Reference: PA No. PA-05-001The National Institute of General Medical Sciences (NIGMS) currently supports the analysis of complex biological systems through investigator-initiated research project grants. The resources needed to conduct the multifaceted, multidisciplinary projects that may be required to achieve significant advances in these complex areas may be beyond the scope of the typical R01 or P01 grant. Therefore, this request for applications (RFA) presents an opportunity for applicants to assemble large teams of investigators from diverse disciplines that may not be possible with other funding mechanisms.Drosophila and Caenorhabditis elegans, and at the clinical level, new approaches are being explored to understand the integrated activity of tissues and organs.The biomedical sciences have undergone a fundamental shift in the conceptual and technical approaches that can be applied to certain problems of profound importance. These problems center on understanding the behavior of biological systems whose function is the product of spatial and temporal ordering of myriad interacting components. Modeling approaches are being used to understand the orderly development of biological pattern in organisms such as Part of the impetus for systems-scale approaches rests on advances in acquiring data of the necessary quality and quantity to permit computational modeling. Among the most striking examples are the availability of complete DNA sequences for hundreds of organisms, including humans, and the availability of high-throughput instrumentation for analyses of gene function such as gene expression microarrays and proteomics technologies. These advances have made it feasible to generate a truly comprehensive parts list for any organism and to track changes over time. Ultimately, it should be possible to enumerate all the informational units of the genomes , their processed forms, and their dynamic presence in cells.Rapid advances in large-scale data collection and analysis have given scientists a global yet detailed view of cellular processes, instead of focusing on individual molecules or a small number of interacting molecules. Unprecedented opportunities have emerged that may open the door to uncover hidden rules governing the ensemble of biomolecules working concertedly to perform certain functions in the cell. In the meantime, substantial challenges in information integration, interpretation, and representation have arisen. In order to move beyond the phase of cataloguing the parts list and truly transform data into knowledge, and knowledge into principles, iterative cycles of data collection and model generation and validation will be necessary.A higher-order problem presents itself in understanding how the genome-encoded components and the other molecules are constituted in networks of interacting molecules with particular distributions in time and space. Advances in imaging techniques and analytic methods are beginning to yield copious quantitative and spatial data on specific molecules in biological systems. Knowledge of the network and changes in its components over time, and the local rules by which the individual components distribute material and information, will substantially advance our knowledge.At the organism level, phenotype must take into account the relationships and interactions of biological and environmental variables. Basic biological systems\u2014including gene sequences, structures, and pathways that direct metabolism and development\u2014vary within individuals, among individuals, among populations, and among species. Advances in complex systems-level understanding must ultimately include models that account for these variations.Medical, biotechnological, and other uses of biological information increasingly depend on our ability to understand the principles and dynamics that explain the behavior of the system as a whole. Whether the goal is to understand the consequences of disease or injury, identify particular molecular targets for drug interventions, or modify the metabolism of microorganisms to produce medicines, the challenge is predictability. Predicting how the system of interest will respond to an intervention is a computational problem. For biological systems, this challenge is daunting.Parallel to scientific challenges are organizational and educational challenges. At the institutional level, building cohesive multidisciplinary research teams by integrating expertise across traditional disciplinary boundaries is not a simple undertaking. Beyond institutions, excessive overlap and redundancy in project selection and tool development exists in the research communities that could be reduced by promoting communications, collaborations, and technology and data sharing. The emergence of new science demands an adequate workforce of new scientists. Training for the future leaders of systems biology research who are knowledgeable and skilled in both experimental and computational subjects is timely. Good mechanisms and plans to address these challenges are significant tasks of the centers.High priority will be given to projects that integrate multi-investigator, multidisciplinary approaches with a high degree of interplay between computational and experimental approaches. Innovation is critical for both research project design and infrastructure design with a mission of serving communities beyond the participating investigators, institutions, and collaborators. A variety of organizational models are possible; it is not the intent of this RFA to prescribe any particular one.http://www.nigms.nih.gov/news/releases/complex_centers.html), two centers in 2003 (http://www.nigms.nih.gov/news/releases/complex_centers-2003.html), and one center in 2004 (http://www.nigms.nih.gov/news/releases/quantitative_bio_center.html). Potential applicants should become familiar with the research focuses of the existing centers. Research conducted by the future centers should complement and enhance projects already funded.The NIGMS awarded two centers under this program in 2002 ; 4) metabolic networks and the control of the flux of substrates, intermediates, and products in cell physiology; 5) organ system networks involved in multiorgan failure in shock, trauma, and burn injury; and 6) genetic architecture of biological complexity related to inherited variation and environmental fluctuations.The NIGMS intends to support systems biology research for the areas that are central to its mission of supporting basic biomedical research, and that focus on developing new computational approaches to biomedical complexity. Research areas that historically have been computationally based are excluded as a focus of this center program. Research focusing on disease processes and their specific organ systems is not eligible. NIGMS mission areas include, but are not limited to, the following: 1) signaling networks and the regulatory dynamics of cellular processes such as cell cycle control, transient complex formation, organelle biogenesis, and intercellular communications; 2) supramolecular machines, such as the replisome, spliceosome, and molecular motor assemblies in cell division and motility; 3) pattern formation and developmental processes in model systems .The NIGMS National Centers for Systems Biology will be expected to provide national leadership in systems biology research and training. To do so, they will be expected to support training and outreach activities that will ensure the flow of information and expertise both into and out of the centers. Centers should have plans to bring the most advanced technologies developed at other laboratories to the centers and to disseminate expertise and knowledge to a wider community through collaborations, visiting investigatorships, fellowships, center websites, workshops, symposia, summer courses/internships, and/or other means. To maximize the impact, centers should conduct training at multiple levels appropriate to their institutions. Incorporation of developmental research projects led by junior and new investigators into the center research and development plans is strongly encouraged. Over a period of time, centers should evolve into integrated research, training, and knowledge exchange headquarters of scientific communities that will be the engines for coordinated scientific discoveries. The centers should also have plans for outreach to undergraduate institutions, including minority-serving institutions. Information on relevant minority-serving institutions may be obtained by consultation with staff of the NIGMS Division of Minority Opportunities in Research .The NIGMS intends to commit up to $7 million in fiscal year 2006 to fund one to three new P50 center grants in response to this RFA. An applicant may request a project period of up to five years and a budget for direct costs of up to $2 million per year, exclusive of subproject fiscal and administrative costs . Applications must have a Dun & Bradstreet (D&B) Data Universal Numbering System number as the universal identifier when applying for federal grants or cooperative agreements. This number can be obtained by calling 1-866-705-5711 or online at http://grants.nih.gov/grants/guide/rfa-files/RFA-GM-05-010.html#PartI.Letters of intent must be received by 25 January 2005, with 25 February 2005 the deadline for applications. The complete version of this announcement is available online at Contact: James J. Anderson, Center for Bioinformatics and Computational Biology, NIGMS, 45 Center Dr, Rm 2As.25A, MSC 6200, Bethesda, MD 20892-6200 USA, 301-594-0943, fax: 301-480-2228, e-mail: andersoj@nigms.nih.gov; Jiayin (Jerry) Li, Center for Bioinformatics and Computational Biology, NIGMS, 45 Center Dr, Rm 2As.19F, MSC 6200, Bethesda, MD 20892-6200 USA, 301-594-0682, fax: 301-480-2004, e-mail: lij@nigms.nih.gov. Reference: RFA No. RFA-GM-03-009"} +{"text": "The Internet has the potential to provide patients and physicians with rapid access to high quality, timely evidence regarding health and medical diagnosis and treatment. However, many barriers must be surmounted before this potential is achieved. Quality of content must be able to be verified, including the accuracy and timeliness of the information, the source of the information, and the objectivity of the source. Advertising and sponsorship must not influence content and should not be juxtaposed with related content. Individuals must be able to access information without loss of personal privacy. To address these issues, the American Medical Association has developed principles to guide development and posting of Web site content, govern acquisition and posting of online advertising and sponsorship, ensure site visitors' and patients' rights to privacy and confidentiality, and provide effective and secure means of e-commerce. While these guidelines were developed specifically for the AMA Web sites and visitors to these sites, they also may be useful to other providers and users of medical information on the Web. Full paper published as:http://jama.ama-assn.org/issues/v283n12/full/jsc00054.html)Guidelines for medical and health information sites on the Internet: principles governing AMA Web sites by Margaret A. Winker; Annette Flanagin; Bonnie Chi-Lum; John White; Karen Andrews; Robert L. Kennett; Catherine D. DeAngelis; Robert A. Musacchio ::1600-1606). ("} +{"text": "The Internet is introducing new ways for humans to interact with machines and to communicate with each other. In health care the Internet is providing unprecedented opportunities to access information, improve decisions, and enhance communication among decision-makers and the people affected by their decisions.However, the Internet is also creating many new problems. Seeking information on the Internet is often time-consuming. Internet users, regardless of their role, background or knowledge, can experience confusion and anxiety because of the virtually unlimited amount of information available, information that is often poorly organized and of highly variable quality and relevance.1 The Internet can also lead to conflict among decision-makers if they have access to different and contradictory information. A person's health might even be worsened if inaccurate information found on the Internet were used by decision-makers.Evidence-based decision-making involves the explicit, conscientious and judicious consideration of the best available evidence in making health care decisions.2 It is supported by a rapidly evolving set of methods and tools but its eventual adoption will depend on whether the barriers it still faces3 can be minimized or eliminated.In this paper we postulate that if the Internet and evidence-based decision-making are to reach their full potential and contribute to improvements in health care, a powerful and efficient synergy must develop between them.4 The Internet could benefit evidence-based decision-making by giving decision-makers cheap, fast and efficient access to up-to-date, valid and relevant knowledge at the right time, at the right place, in the right amount and in the right format. Conversely, the tools and principles of evidence-based medicine could be used to gain a better understanding of the role of the Internet in health care, helping us to anticipate opportunities and prevent potential problems.This article briefly describes some of the efforts that are already fostering convergence and synergy between the Internet and evidence-based decision-making, as well as the opportunities available and the challenges to be overcome. Full paper published as:http://www.cma.ca/cmaj/vol-162/issue-3/0362.htm)A. R. Jadad, R. B. Haynes, D. L. Hunt, and G. P. Browman. The Internet and evidence-based decision-making: a needed synergy for efficient knowledge management in health care. CMAJ 162 (3):362-365, 2000. ("} +{"text": "An essential element of the mission of the NIEHS is the support and career promotion of the future generation of exceptionally talented and creative new scientists who will further the understanding of the impact of environmental exposures on human health. The NIEHS supports a number of training and fellowship programs for pre- and postdoctoral training, and mentored career development awards for faculty in the early stages of their career development. Primary among these are the Ruth Kirschstein National Research Service Awards for pre- and postdoctoral training, the Career Development Awards for clinically trained scientists (K08 and K23), and the Mentored Quantitative Research Career Development Awards to support the career development of scientists with quantitative and engineering backgrounds who wish to integrate their expertise with biomedicine. In addition, in 1999 the NIEHS instituted the Transition to Independent Positions Program to address the progression of individuals from postdoctoral positions to faculty positions. In this career development award the individual applies for the grant while still in a postdoctoral position, and the grant for start-up funding is awarded at the institution where the candidate accepts the faculty position. However, even with these career development mechanisms in place, to fulfill its mission of assuring a cadre of productive environmental health science investigators for the future, NIEHS needs to initiate further imaginative programs to identify the best new biomedical investigators and facilitate their establishing vibrant, independent research programs in the environmental health sciences.To identify outstanding scientists at the formative stages of their career and assist them in launching an innovative research program with a defined impact in the environmental health sciences, the NIEHS is establishing a program of R01 research grants intended for researchers who have not received their first R01 research grant. It is designed to be highly competitive, and only a limited number will be awarded per year.Research programs supported by this announcement seek to promote career advancement of the most highly creative and promising new scientists who intend to make a long-term career commitment to research in the mainstream of the environmental health sciences, and bring innovative, ground-breaking research initiatives and thinking to bear on the problems of how environmental exposures affect human biology, human pathophysiology, and human disease.The R01 applications in this program are distinguished from other R01 research grants in that the applications 1) incorporate a statement of career goals in the environmental health sciences; 2) include a discussion of previous research experience and achievements in addition to the research proposal; 3) may include active participation of an external advisory committee; 4) require demonstration of the commitment by the institution to actively support the research program development of the principal investigator (PI); and 5) include a separate budget specifically devoted to equipment and career enhancement activities.Research projects proposed in response to this Request for Applications will be expected to have a defined impact on the environmental health sciences and be responsive to the mission of the NIEHS, which is distinguished from that of other Institutes by its focus on research programs seeking to link the effects of environmental exposures to the cause, mechanisms, moderation, or prevention of a human disease or disorder or relevant pathophysiologic process. For purposes of this announcement, all applications must focus on a specific human disease, dysfunction, pathophysiologic condition, or relevant human biologic process and propose to study a specific environmentally relevant toxicant. Examples of environmentally relevant toxicants include industrial chemicals or manufacturing byproducts, metals, pesticides, herbicides, air pollutants, and other inhaled toxicants, particulates or fibers, and fungal, bacterial, or biologically derived toxins. Agents considered nonresponsive include, but are not limited to, alcohol, chemotherapeutic agents, radiation that is not a result of an ambient environmental exposure, drugs of abuse, pharmaceuticals, and infectious or parasitic agents, except when these are disease co-factors to an environmental toxicant exposure to produce the biological effect.Applicants involving animal exposures must include a justification of how the exposure paradigm is relevant to human exposure and clearly discuss the link between the exposure and the relevant human disease in the Background and Significance section of the application. The applicant should also discuss the potential for translating the research\u2014applying the ideas, insights, and discoveries generated through the basic inquiry to the treatment or prevention of human disease. Applicants proposing epidemiologic research should address how the significant associations revealed in the studies could be confirmed in the laboratory setting.The ONES program would be evaluated on a continuing basis by the NIEHS, to assess the impact of the program on the portfolio of the NIEHS as well as on the progression of the awardees' careers. Metrics to be used include, but are not limited to, publications (numbers and impact factors of publications); academic promotion of PIs; awards, invited talks at national/international symposia, students and postdoctorals trained in the PI's laboratory, and honors received by PIs; committee service by PIs; and subsequent grant support awarded. The design of the program evaluation will be determined by the Program Analysis Branch of the Division of Extramural Research and Training. PIs of awarded ONES grants must provide information for the evaluation and any subsequent program evaluations for up to 10 years after the award.http://grants.nih.gov/grants/funding/phs398/phs398.html). A detailed categorical budget for the \"Initial Budget Period\" and the \"Entire Proposed Period of Support\" is to be submitted with the application. For further assistance contact GrantsInfo, 301-435-0714 (telecommunications for the hearing impaired: TTY 301-451-0088) or by e-mail: GrantsInfo@nih.govThis funding opportunity will use the R01 Grant mechanism. This funding opportunity uses the just-intime budget concepts. It also uses the nonmodular budget format described in the PHS 398 application instructions Data Universal Numbering System number as the universal identifier when applying for federal grants or cooperative agreements. The D&B number can be obtained by calling 866-705-5711 or through the web site at http://grants.nih.gov/grants/guide/rfa-files/RFA-ES-05-005.htmlThe deadline for receipt of letters of intent is 20 November 2005, with 22 December 2005 the deadline for receipt of applications. The complete version of the RFA is available at shreffl1@niehs.nih.gov. Reference: RFA-ES-05-005Contact: Carol Shreffler, Division of Extramural Research and Training, National Institute of Environmental Health Sciences, P.O. Box 12233, EC-23, 111 T.W. Alexander Drive, Research Triangle Park, NC 27709 USA, 919-541-1445, fax: 919-541-5064, e-mail: This initiative continues NIEHS\u2019s and EPA\u2019s intent to foster advances in children\u2019s health by supporting innovative, state-of-the-art Research Centers examining the adverse health effects of environmental exposures among children. Both agencies are interested in reducing environmental risks to children and alleviating the societal burden of environmentally-induced disease/dysfunction.Collaborative, multidisciplinary research approaches are required to explore the dynamic interaction of children and the environment. Centers are expected to have fully coordinated programs that incorporate exposure and health effects research to support the development and validations of novel health promotion strategies. A Center should identify a central theme or focus of its research effort so that the proposed projects are responsive to the specific research area of children\u2019s environmental health included in this Center program.The program emphasizes integration of basic and population sciences while utilizing a community-based participatory research (CBPR) approach. By bridging gaps between basic and clinical research and between institutional researchers and community members, this program aims to improve our knowledge regarding detection, treatment, and prevention of environmentally-induced disease/dysfunction in children.The long-range goals of this program are 1) to stimulate new and expand existing research on the role of environment in the etiology of disease/dysfunction among children, 2) to develop novel effective intervention and prevention strategies, and 3) to promote translation of basic research findings into applied intervention and prevention methods, thereby enhancing awareness among children, their families, and health care practitioners regarding detection, treatment, and prevention of environmentally related diseases and health conditions.A spectrum of scientific approaches is expected to create a truly multidisciplinary working environment where basic research can inform clinical research. These may include: 1) mechanistic research including pathophysiology of target-organ system; 2) toxicologic research; 3) molecular and cellular sciences; 4) clinical research; 5) public health research including epidemiology; 6) exposure assessment and remediation; 7) behavioral and social sciences; 8) cost/benefit; and 9) social policy research.Each Center must produce a synergistic research environment that allows each research effort to share the creative strengths of the others. Each Center, by supporting interrelated projects and collaborating investigators, is expected to yield results beyond those achievable were each project pursued separately and without formal interaction among the participating investigators. The demonstration of synergy among the projects and the multidisciplinary nature of the work are critical components of this program. Ultimately, the expected outcomes of this research program include 1) the generation of cutting-edge science, 2) the development of local and national networks of children\u2019s environmental health research professionals, and 3) peer-reviewed data relevant to better understanding vulnerability and risk. Expected outputs from each Center include 1) contributions to the scientific literature and 2) translational outreach and communication tools developed with and applicable for the effected community of concern.in utero exposure. This includes any endocrine active chemical(s) or organic solvents, particulate matter (PM), pesticides, nutritional supplements, phytochemicals or metals; nutrition and social and cultural factors cannot be considered alone. Applicants are encouraged, however, to incorporate these factors in assessing the effects of previously described environmental exposures. These areas are of interest: 1) mental retardation, cerebral palsy, autism spectrum disorders, visual and or hearing impairment, attention deficit and hyperactivity disorders, and affective disorders; 2) delays or deficits in domains of neurodevelopment such as cognition, motor, sensory; 3) thyroid/pituitary dysfunction, puberty and sexual maturation, reproductive development, sexual dimorphic phenotypes, growth disorders or impairments including but not limited to childhood obesity; 4) reproductive outcomes such as preterm delivery, birth defects; 5) other outcomes associated with nervous and/or endocrine system disruption.For this RFA, NIEHS and EPA will accept applications focused on environmentally mediated disorders/dysfunctions of the nervous and/or endocrine systems. Applicants must study an environmental agent/chemical/stressor to which there is human exposure and the potential for Each Center will propose an overall research mission and plan that is responsive to the objectives of the NIH Center Program. The application must contain a minimum of three research projects, including the first two listed below: 1) laboratory basic research project; 2) clinical research project; 3) other research project(s). At least one of the three projects must use the CBPR process as defined below. Applications lacking the first two projects and a project using the CBPR process will be considered nonresponsive and returned without review.Each Center must also include the Community Outreach and Translation Core (COTC) and the Administrative Core. Applications lacking these two cores will be considered nonresponsive and returned without review.in vitro systems, and/or human clinical specimens.Laboratory-based research projects may include mechanistic studies of environmental agents that contribute to adverse health outcomes in children as well as research that will improve our basic understanding of pathophysiology, molecular genetics, or cell biology of developmental processes. Basic mechanistic research may pertain to the disciplines of toxicology, cell and molecular biology, physiology, psychology, genetics, or other relevant fields, and methods may include animal models, in vitro studies that utilize human tissues that cannot be linked to a living individual. For this RFA, two types of clinical research would be accepted: 1) patient-oriented research ; 2) epidemiologic and behavioral studies. Clinical research that explores gene and environmental interactions in risk of disease/dysfunction is highly encouragedClinical research is conducted with human subjects for which an investigator (or colleague) directly interacts with human subjects. Excluded are At least one other \u201cresearch project(s)\u201d must be proposed. The additional project(s) must be thematically related and integrated with the above two projects. For example: 1) Studies characterizing pathways of exposure; the magnitude, frequency, duration, and time-pattern activities that lead to contact in children and quantifying contact rates of children with exposure media, contaminant transfer efficiencies, and uptake rates in children; 2) research on behavioral factors that affect exposure and or the ability to reduce exposure; 3) research that characterizes the economic and social impact of children\u2019s illnesses on society; 4) evaluation of how scientific information about children\u2019s environmental health affects policy, social change, and changes in clinical and public health management of these diseases.In CBPR, scientific inquiry is such that community members, persons affected by the health condition, disability, or issue under study, or other key stakeholders in the community\u2019s health can be full participants in each phase of the work . CBPR is characterized by substantial community input in the development of the project.Community refers to populations that may be defined by geography; race; ethnicity; sex/gender; sexual orientation; disability, illness, or other health condition; or to groups that have a common interest or cause, such as health or service agencies and organizations, health care or public health practitioners or providers, policy makers, or lay public groups with public health concerns. Community-based organizations may be involved in the research process as members or representatives of the community. Organizations as varied as tribal governments and colleges, state or local governments, independent living centers, other educational institutions such as junior colleges, advocacy organizations, health delivery organizations , health professional associations, nongovernmental organizations, and federally qualified health centers are possible community partners. Each Center must include 1) Description of Cores and 2) a COTC and an Administrative Core.A COTC is required to develop, implement, and evaluate strategies to translate and apply the scientific findings of the Center into information for the public, policy makers, and clinical professionals to use to protect the health of children. This must include personnel from one or more of the following areas: health educators, nurses, members of community or faith-based organizations, members of organizations that advocate for research and services pertaining to children\u2019s health, members of professional societies of health care professionals, and state and local health departments or medical service organizations. Examples of activities considered responsive are creating training materials for health professions, developing new ways to disseminate research findings to the broad audience of stakeholders, and assessing community understanding of research results and plans for action. A Center will devote at least 10% of its budget to the COTC.Each Center must include an Administrative Core unit to provide oversight, coordination, and integration of Center activities and establish an External Advisory Committee (EAC) to the Center Director. The EAC should consist of three to five scientists with expertise appropriate for the Center\u2019s research focus, plus one representative from a community-based organization involved in community-based research. Representation from a state or local health department is also encouraged. At least 67% of Committee members should be from outside the grantee institution. The membership of the EAC must be approved by the funding agency. The EAC is to help evaluate the merit, value, and contribution of research projects and the relevance and importance of individual organizational elements to the overall goals of the Center. The membership of the EAC must be approved by the Participating Agencies postreview; names should not be submitted in the application. Individuals in senior leadership positions should provide intellectual, administrative, and scientific leadership for the Center and are critical to its overall effectiveness and evolution. These individuals should be in place and committed to a defined percent effort. Please submit only a description of proposed protocols and planned committee by representation and area of expertise. If awarded, you must provide an identifiable list of membership to the EAC for approval by the funding agencies.Each Center may support other cores that provide a technique, service, or instrumentation that will enhance ongoing research efforts, such as animal resources, cell/tissue culture, pathology, biostatistics, molecular biology, neuropsychology, neuroimaging, analytical chemistry, exposure assessment, genotyping, and resequencing. Budgeted Center projects and external research projects may have access to these cores. The application should provide a total operational budget for each facility core together with the percentage of support requested from the Center grant. The application should explain the organization and proposed mode of operation of each core, including a plan for usage, priority setting, allocation of resources, and any applicable charge-back system.Within the Center Program, PIs are encouraged to support training of new investigators within the proposed projects. New investigators should have a doctoral degree with < 8 years of postdoctoral experience at the time of application, and have demonstrated outstanding abilities in basic, clinical, or population-based research, such as postdoctoral research in an academic, industry, or government environment. However, years of clinical training will not count against the limitation. Ineligible individuals include current and former PIs on EPA STAR Grants or NIH research projects (R01), subprojects of program projects (P01), or Center Grants with research components (P50) or equivalent research grant awards. Applicants will be expected to devote at least 50% time and effort to the award and have a long-term commitment to research in the environmental health sciences.This funding opportunity will use the NIH P01 award mechanism and the EPA\u2019s Office of Research and Development, STAR Grant awards. As an applicant, you will be solely responsible for planning, directing, and executing the proposed project.http://www.dnb.com/us/. The D&B number should be entered on line 11 of the face page of the PHS 398 form.Applications must be prepared using the most current PHS 398 research grant application instructions and forms. Applications must have a D&B Data Universal Numbering System (DUNS) number as the universal identifier when applying for Federal grants or cooperative agreements. The D&B number can be obtained by calling (866) 705-5711 or through the web site at http://grants.nih.gov/grants/funding/phs398/phs398.html). A detailed categorical budget for the \u201cInitial Budget Period\u201d and the \u201cEntire Proposed Period of Support\u201d is to be submitted with the application. For further assistance contact GrantsInfo, 301-435-0714 (telecommunications for the hearing impaired: TTY 301-451-0088) for by e-mail: GrantsInfo@nih.gov.This funding opportunity uses the just-in-time budget concepts. It also uses the non-modular budget format described in the PHS 398 application instructions \u00aasee http://grants.nih.gov/grants/guide/rfa-files/RFA-ES-05-004.html.The deadline for receipt of letters of intent is October 23, 2005, with November 24, 2005 the deadline for receipt of applications The complete version of the RFA is available at kg89o@nih.gov; Chris Saint, National Center for Environmental Research, U.S. Environmental Protection Agency (8723R), 1200 Pennsylvania Ave. NW, Washington, DC 20460 USA, 202-564-6909, fax: 202-565-2448, e-mail: saint.chris@epa.gov; Nigel Fields, National Center for Environmental Research, U.S. Environmental Protection Agency (8723F), 1200 Pennsylvania, Ave. NW, Washington, DC 20460 USA, 202-343-9767, fax: 202-233-0677, e-mail: fields.Nigel@epa.gov. Reference: RFA-ES-05-004Contact: Kimberly A. Gray, Division of Extramural Training and Science National Institute of Environmental Health Sciences, PO Box 12233, EC-21, 111 T.W. Alexander Drive, Research Triangle Park, NC, 27709 USA, 919-541-0293, fax: 919-316-4606, e-mail:"} +{"text": "Education in bioinformatics has undergone a sea change, from informal workshops and training courses to structured certificate, diploma, and degree programs\u2014spanning casual self-enriching courses all the way to doctorate programs. The evolution of curriculum, instructional methodologies, and initiatives supporting the dissemination of bioinformatics is presented here.Building on the early applications of informatics (computer science) to the field of biology, bioinformatics research entails input from the diverse disciplines of mathematics and statistics, physics and chemistry, and medicine and pharmacology. Providing education in bioinformatics is challenging from this multidisciplinary perspective, and represents short- and long-term efforts directed at casual and dedicated learners in academic and industrial environments. This is an NP-hard problem.Training in bioinformatics remains the oldest and most important rapid induction approach to learning bioinformatics skills. Both formal (short-term courses) and informal training (on-demand \u201chow-to\u201d procedures) have remained the mainstay of on-the-job programs.After almost a decade of short-term training, and retraining students, faculty, and scientists in discrete aspects of bioinformatics, the impetus to formalize bioinformatics education came in 1998 from Russ Altman [http://surya.bic.nus.edu.sg/web01/) was launched at the 2001 International Conference on Intelligent Systems for Molecular Biology (ISMB) as a satellite meeting, to provide for the first time a forum for bioinformatics educators to meet, discuss, and exchange ideas and suggestions. WEB addresses fundamental educational and pedagogical issues to determine the nature, extent, and content of, and delivery tools available for, bioinformatics degree and training programs, and to provide focus points and suggestions for improvement of nascent degree programs.The Workshop on Education in Bioinformatics (WEB) 2001 on the opening day of the conference.How well do the new graduates from these educational programs fit the jobs available to them? How many areas should a bioinformatics graduate be an expert in? Apart from a deep understanding of algorithms, programming, and life sciences, solving problems in genomics, proteomics, and/or medical informatics appears to be the current requirement. With the blurring boundaries between bioinformatics and new areas of endeavor such as forensics and biodiversity/ecoinformatics, where should we, as educators, draw the line? With industry's annual fluctuating demands for specific in-depth knowledge, how can we create a bioinformatics world?Some of these issues will be discussed in the 2006 WEB meeting. But here are four take home messages from the earlier workshops. First, with the growing demand for computational biologists, there is a persistent and continuing demand for bioinformatics education at all levels\u2014formal and informal, face-to-face and distance learning, and short-term training and rigorous long-term academic programs. Clearly, \u201cone size does not fit all\u201d , since t"} +{"text": "This program is intended to promote productive development of foreign investigators from low- to middle-income countries, trained in the United States or in their home countries through specific Fogarty International Center D43 or U2R training programs, to independent researcher in their home countries or other low- or middle-income countries. This program will dovetail with several specific research training mechanisms, including training in the NIH Intramural Visiting Fellows Program, the Fogarty International Center research training programs (D43 or U2R), the NIDA INVEST, or Humphrey Fellowships, NIEHS junior scientist programs, and the Human Frontier Science Program, as part of a broader program to enhance the scientific research infrastructure in low- to middle-income countries, to stimulate research on a wide variety of high-priority health-related issues of importance in those countries, and to advance NIH efforts to address health issues of global import.The specific goal of this initiative is to provide funding opportunities for the increasing pool of foreign social and behavioral scientists, clinical investigators, nurses, and other health professionals, on their return to their home countries, with state-of-the-art knowledge of research methods to advance the critical issues in global health through social and behavioral sciences research.After their term of research training, low- to middle-income country participants supported by this announcement are expected to continue independent and productive scientific careers, including providing expert training and consultation to others, and/or research on behavioral and/or social science issues within their home institutions.This announcement contributes to the FIC mission and the broad NIH initiative to reduce health disparities among nations by strengthening research infrastructure in low- to middle-income countries, particularly those with the least economic resources. It also provides the opportunity for recently trained international health and health care researchers to continue their projects after returning home.Definitions of \u201cbehavioral\u201d and \u201csocial\u201d: For the purposes of this initiative, the term \u201cbehavioral\u201d refers to overt actions; to underlying psychological processes such as cognitions, emotion, temperament, and motivation; and to biobehavioral interactions. The term \u201csocial\u201d encompasses sociocultural, socioeconomic, and sociodemographic status; to biosocial interactions; and to the various levels of social context from small groups to complex cultural systems and societal influences.The core areas of behavioral and social sciences research are those that have a major and explicit focus on the understanding of behavior and social processes, or on the use of these processes to predict or influence health outcomes or health risk factors. These core areas of research are divided into basic research and clinical research.http://www.drugabuse.gov/International/HHHRF.html) plus at least one additional year of significant, well-documented, mentored research; 4) at least 2 years of research training experience through the NIH Intramural Visiting Fellows Program; 5) 1 year of training through an F05 international fellowship program and 1 subsequent year of mentored research; 6) recipients of Long-Term Fellowship awards through the Human Frontier Science Program, who come from low- and middle-income countries, and who have spent at least 2 years in research training; 7) at least 1 year of training in the United States and one additional year of significant, well-documented, mentored research, in the United States or abroad, leading to a completed master\u2019s degree or doctoral degree, at least partially funded through a Fogarty International Center research training program, with preapproval by the program director; 8) foreign trainee researchers from low- or middle-income countries trained under NIEHS R01, R37, and P01 programs, as described at http://www.niehs.nihgov/dert/programs/capacity.htm.As part of its global health initiative under the Department of Health and Human Services (DHHS), the Fogarty International Center (FIC), in partnership with the National Eye Institute (NEI); the National Institute on Drug Abuse (NIDA); the National Institute of Environmental Health Sciences (NIEHS); the National Institute of General Medical Sciences (NIGMS); all of the National Institutes of Health (NIH), and the Office of Behavioral and Social Sciences Research (OBSSR), the Office of Dietary Supplements (ODS), and the Office of Research on Women\u2019s Health (ORWH), all of the Office of the Director (OD) of the NIH, invites applications from current and former NIH-supported foreign research trainees to compete for funds that will support their research efforts upon return to their home countries. To be eligible, foreign scientists must meet at least one of the following criteria: 1) at least 2 years of research training experience under an FIC-supported training grant (classified by the D43 and U2R mechanisms); 2) 1 year of such D43 or U2R training experience coupled with 1 year of significant, well-documented, mentored research experience Grant (IRID-NIAID) program); 3) 1 year of the NIDA INVEST or Humphrey Fellowship provides the opportunity for junior foreign scientists to compete for such funds through a peer-reviewed process. This is a critical adjunct in the continuation of promising independent research careers that will be of benefit to the investigators\u2019 home countries and the world at large. Women and underrepresented minority scientists in their countries are especially encouraged to apply for these re-entry grants. Project proposals should be geared toward the research interests of the applicant and the focus on high-priority health and health care problems in the investigator\u2019s home country that also carry global importance, and are of interest to the collaborating NIH Institutes, Centers, and Offices (ICs) listed on the first page of this announcement, as well.http://www.hhs.gov/ohrp/) that outlines these regulations. For information on animal protection in research, see http://grants.nih.gov/grants/olaw/olaw.htm.It is expected that research topics will vary. Specific research interests of partnering ICs can be found on the ICs\u2019 websites, as listed in the beginning of this announcement. Research related to women\u2019s health, including studies of gender differences in disease onset and progression, identification of behavioral strategies that are effective in encouraging healthy lifestyles in young girls and women, as well as behavioral strategies to encourage prevention of diseases such as STDs and diseases with higher prevalence among women , are particularly encouraged. Research on healthy outcomes of pregnancy and child survival, and population research as associated with both behavioral and social, and economic research is encouraged. Research related to the health effects of human exposures to environmental agents is encouraged. Research focused on behavioral and social determinants and their effects on health is also encouraged. All research must be performed in accordance with NIH and U.S. government regulations regarding the responsible conduct of research. This announcement precludes the support of research involving enrollment in pilot studies for clinical trials, or the actual support of clinical trials since the resources and infrastructure to support and oversee such trials generally exceed the resources available under this award mechanism. Applicants are encouraged to visit the website of the Office of Human Research Protection (OHRP) development of the laboratory capabilities or research projects; 2) training of other potential researchers; 3) publications in international peer-reviewed journals, as well as local journals; 4) participation in workshops, seminars, and international conferences; 5) collaborations with past mentors, as well as other U.S., international, or local researchers; and 6) attraction of funding from other sources.As part of its assessment of the impact and scientific productivity of this program, FIC plans to track researchers and their trainees for at least 10 years after beginning their independent research. Evaluation may focus on the success of researchers , their sustained commitment to research careers, their ability to attract funding for their work, their contributions to future international collaborations, their influence on the development of scientific research in their countries, and their ability to act as consultants, teachers, and role models to other local investigators and further disseminate the lessons learned. Applicants should describe potential metrics that would indicate the success of the individual researcher and the success in capacity building at the home institution, including the impact of the program on research at the institution in the home countries of researchers and their trainees.This Funding Opportunity Announcement (FOA) will use the NIH Research Project Grant (R01) award mechanism.The applicant will be solely responsible for planning, directing, and executing the proposed project.http://grants.nih.gov/grants/funding/modular/modular.htm).This FOA uses \u201cJust-in-Time\u201d information concepts. It also uses the nonmodular budget formats application forms and the SF424 (R&R) Application Guide for this FOA through Grants.gov/Apply.Note: Only the forms package directly attached to a specific FOA can be used. You will not be able to use any other SF424 (R&R) forms , although some of the \u201cAttachment\u201d files may be useable for more than one FOA.GrantsInfo@nih.gov.For further assistance, contact GrantsInfo, 301-435-0714, (telecommunications for the hearing impaired: TTY 301-451-0088), or by e-mail:The letter of intent dates for this PAR are 21 August 2007, 2008, and 2009 with the application receipt dates 21 September 2007, 22 September 2008, 21 September 2009.www.grants/guide/pa-files/PAR-07-328.html.The complete version of this PAR is available at http//http://grants.nih.gov/grants/guide/pa-files/PAR-07-328.html.The complete list of agency contacts is available at Reference: PAR-07-328.The goal of this FOA is to increase understanding of the transition of microbes from a nonpathogenic state to a state pathogenic for humans.One area of interest is to promote research in emerging zoonotic viruses, which will provide predictive tools to anticipate the emergence of new viral pathogens. Many emerging viral diseases are zoonotic in origin; it is estimated that approximately 62% of human infectious diseases are zoonotic and an estimated 70% of newly emerging diseases are zoonotic in origin. The recent emergence and transmission to humans of SARS CoV from civet cats and deadly avian influenza clearly demonstrate that emerging zoonotic viral diseases are a serious public health problem that needs to be understood fully. Although it is clear that one of the principle mechanisms involved in the emergence of new viral diseases is a change in the host range, there are many gaps in our knowledge about the mechanism(s) involved and how zoonotic viruses establish themselves in humans. An understanding of these processes will be necessary to anticipate and prepare for the emergence of new viruses in humans.Staphylococcus aureus colonizes skin or mucosal tissues without causing disease or when Clostridium difficile colonizes the gut without causing illness or when Neisseria meningitidis is in the nasopharyngeal tract without causing disease. Little if anything is known about the full complement of microbes present or the structure and functioning of the microbial consortium. It is important to know what microbes are present, in what proportions, and how the consortium functions metabolically. Clearly, the consortium has an interface with the host and this may play a critical role in determining if the microbes remain as colonizers or cause disease. There are several potential colonizing niches in the body for which this concept applies such as skin, gastrointestinal, nasopharyngeal, and genitourinary tracts. Metagenomics as well as microscopic techniques that allow for visualization of the physical structure of the microbial consortium are experimental approaches of interest.Another area of interest is the study of human bacterial pathogens that exist in colonizing, rather than infective states. In colonizing states these human pathogens are in consortia with other microbes, such as when Two recent workshops , sponsored in part or in whole by NIAID, resulted in reports that identify areas that are in need of additional research. This FOA is intended to address some of these areas.Relevant topics of interest include, but are not limited to: 1) Studies on the natural history of emerging viruses, viral reservoirs, natural means of transmission, variation and pathogenesis ; 2) Studies on the factors that control host\u2013host transmission: pathogenesis studies focused on the development of emerging human pathogenic viruses; viral and host control of viral shedding and infection; 3) Studies on the evolutionary events surrounding the transfer of viruses to new hosts, sequence analysis of viruses before and after the transfer, emergence of variants in the donor host, the transfer event, and post-transfer adaptation; 4) Characterizing human microbial pathogens as they live in nonpathogenic microbial consortia. Research should focus on describing what microbes are present in the consortia, and the metabolism of the consortia, their physical structure and interaction with the environment. Examples of possible experimental approaches include metagenomics, single-cell analysis, and microscopy; 5) Understanding the relationship between microbiota and human health as it influences emergence of disease or new human pathogens; 6) Identifying signals and systems in bacterial communication relevant to the development of pathogenesis or emergence of new human pathogens; 7) Determining whether and how microbiota and human pathogens communicate with host cells and respond to the mammalian immune system; 8) Developing strategies to manipulate chemical signaling of human pathogens within the microbiota.This FOA will not support studies on HIV or pathogens among the NIAID Category A, B & C Priority Pathogens.This Funding Opportunity Announcement (FOA) will use the NIH Research Project Grant (R01) award mechanism.The applicant will be solely responsible for planning, directing, and executing the proposed project.http://grants.nih.gov/grants/funding/modular/modular.htm).This FOA uses \u201cJust-in-Time\u201d information concepts. It also uses the modular as well as the nonmodular budget formats , use the PHS398 Modular Budget component provided in the SF424 (R&R) Application Package and SF424 (R&R) Application Guide .U.S. applicants requesting more than $250,000 in annual direct costs and all foreign applicants must complete and submit budget requests using the Research & Related Budget component found in the application package for this FOA. See NOT-OD-06-096, 23 August 2006.Applicants must download the SF424 (R&R) application forms and the SF424 (R&R) Application Guide for this FOA through Grants.gov/Apply.Note: Only the forms package directly attached to a specific FOA can be used. You will not be able to use any other SF424 (R&R) forms , although some of the \u201cAttachment\u201d files may be useable for more than one FOA.GrantsInfo@nih.gov.For further assistance, contact GrantsInfo, 301-435-0714, (telecommunications for the hearing impaired: TTY 301-451-0088), or by e-mail:http://grants1.nih.gov/grants/funding/submissionschedule.htm.The application submission dates of this PA are available at http://grants.nih.gov/grants/guide/pa-files/PA-07-246.html.The complete version of this PA is available at petersn@niaid.nih.gov; Eun-Chung Park, Division of Microbiology and Infectious Diseases, National Institute for Allergy and Infectious Diseases, Room 4103, MSC-6604, 6610 Rockledge Drive, Bethesda, MD 20892-6604 USA, 301-402-0071, fax: 301-402-2508, e-mail:epark@niaid.nih.gov; Irene Anne Eckstrand, National Institute of General Medical Sciences, 45 Center Drive, Room 2AS.25K, MSC 6200, Bethesda, MD 20892 USA, 301-594-0943, e-mail:eckstrai@mail.nih.gov.Contacts: N. Kent Peters, Division of Microbiology and Infectious Diseases, National Institute for Allergy and Infectious Diseases, Room 4068, MSC-6603, 6610 Rockledge Drive, Bethesda, MD 20892-6603 USA, 301-496-7728, fax: 301-402-2508, e-mail:Reference: PA-07-246."} +{"text": "The first time I heard the term \"e-health\" I was at the 7th International Congress on Telemedicine and Telecare in London, at the end of November 1999. John Mitchell from Sidney, Australia, spoke about a national government study whose main result was the recognition that \"cost-effectiveness of telemedicine and telehealth improves considerably when they are part of an integrated use of telecommunications and information technology in the health sector.\" .In this talk, e-health was introduced as the death of telemedicine, because - in the context of a broad availability of medical information systems that can interconnect and communicate - telemedicine will no longer exist as a specific field. The same could also be said for any other traditional field in medical informatics, including information systems and electronic patient records. e-health presents itself as a common name for all such technological fields.Mitchell also pointed out that \"e-health can be considered to be the health industry's equivalent of e-commerce,\" and this could be one key for understanding the sense of e-health: just medical informatics and telematics on the shop shelves, a fashionable name for something already existing but otherwise difficult to sell.Telemedicine Today- a non-peer-reviewed journal - changed from \"Where healthcare + telecommunications converge\" to \"The eHealth Newsmagazine,\" and just some months later, even the Telemedicine Journal- a scientific, peer-reviewed journal - added an \"and eHealth\" to its title. Nice name? Fear of being left out of a possibly-new field? The Ace Allen editorial that introduces the change in the subtitle of Telemedicine Today [Telemedicine Today's name suddenly changed, perhaps to satisfy the hundreds of healthcare-related dotcoms looking for a buzzword.Without arguing anything about the consequentiality of the facts, in December 1999 the subtitle of ne Today sounds sShortly after the above changes, E. Rosen rationalAllen, in a further editorial , discoveTelemedicine Journal and eHealth did not publish any paper directly mentioning e-health; also, the other major scientific journal related to telemedicine, the Journal of Telemedicine and Telecare, seems not to care much about e-health. This could be related to the business role the term e-health seems to have: when researchers describe their work, the classical categories of, for example, Medical Informatics, Telemedicine, and Electronic Patient Records are more meaningful than the generic term e-health.Interestingly enough, even after the name change, As a researcher, I can see some sense in the term e-health; coming from the integration perspective it suggests: integrated-healthcare-systems' properties, possibilities, and consequences that are (in a holistic approach) more than the sum of the single-component outcomes. However, even these aspects are already studied in some computer science fields - for example Artificial Intelligence (at least inside the multi-agent paradigm), Information Economics, and Dynamic Systems; thus, there is nothing new again, except for the specific interest in healthcare.Vincenzo Della Mea Institute of Pathology, University of Udine, Italy"} +{"text": "The Japan Society for the Promotion of Science (JSPS) conducts fellowship programs for foreign researchers to promote international cooperation in and mutual understanding through scientific research in Japan. These programs provide opportunities for U.S. researchers to conduct cooperative research under their host researchers. Research applications are accepted at the John E. Fogarty International Center of the National Institutes of Health, which acts as a nominating authority for JSPS programs. As the eligibility requirements and application procedures are different for each fellowship, the following information describing eligibility and application procedures should be reviewed before submitting an application.The JSPS conducts short- and long-term programs under the Invitation Fellowship Program, funded by a subsidy from the Japanese government. The fellowship allows scientists employed at designated Japanese research institutions and laboratories to invite fellow researchers from the United States to Japan to participate in cooperative activities. These visits presuppose the existence of contacts between scientists in Japan and other countries, a condition considered favorable to the promotion of future scientific cooperation and exchange. Application deadline: 31 October 2005.The JSPS conducts short- and long-term Postdoctoral Fellowships for Foreign Researchers. These fellowships were established to assist promising and highly qualified young researchers wishing to conduct research in Japan. The program is aimed at providing opportunities for such researchers to, under the guidance of their hosts, conduct cooperative research with leading research groups in Japanese universities and other institutions, thereby permitting them to advance their own research while stimulating Japanese academic circles\u2014particularly young Japanese researchers\u2014through close collaboration in scientific activities. The program\u2019s intention is also for such collaboration to advance scientific research in the counterpart countries. Postdoctoral Fellowship (short-term) application deadline: 31 October 2005. Postdoctoral Fellowship (long-term) application deadline: 30 June 2005.ferreima@mail.nih.gov.Contact: Maria \u201cMili\u201d Ferreira, Division of International Training and Research, Fogarty International Center, NIH, 31 Center Dr, Bldg 31, Rm B2C39, MSC 2220, Bethesda, MD 20892-2220 USA, 301-594-9778, fax: 301-402-0779, e-mail:The National Academy of Engineering (NAE), supported by The Grainger Foundation, has established the Grainger Challenge Prize for Sustainability. The primary purpose of this \u201cinducement prize\u201d is to accelerate the development and dissemination of technologies to enhance social and environmental sustainability for the benefit of current and future generations. A complementary goal of the prize competition is to increase awareness among the U.S. engineering community of the importance of designing and engineering for sustainability, particularly in an international context, and to encourage and showcase efforts by U.S. engineers to bring sustainable technologies to the marketplace and promote green design philosophies. The recipient(s) of the prize will be awarded US$1 million.The specific goal of this competition, which may be followed by future prize competitions in like amounts for comparable goals, will be the development of a community- or household-scale water treatment system to remove arsenic from the contaminated groundwater found in many developing countries. The system must have a low life-cycle cost; must be technically robust, reliable, maintainable, socially acceptable, and affordable; must be manufactured and serviced in a developing country; and must not degrade other water quality characteristics.Arsenic contamination has affected millions of people, primarily in rural Bangladesh, as well as in eastern India, Nepal, and several other countries. In Bangladesh, the arsenic is an unintended consequence of an aggressive international program to control the spread of cholera spread through surface waters by drilling thousands of tubewells. Unfortunately, the tubewells tapped into aquifers containing hundreds of parts per billion (ppb) of naturally occurring arsenic, usually within 100 meters of the surface.Efforts to solve this problem have been under way for a decade, but no single solution has been implemented on a large scale. Field and laboratory tests have been conducted on technologies to determine if they are affordable, robust, and meet World Health Organization (WHO) water quality standards for a treatment system that can be used either in individual homes or at the community level. The intent of the NAE/Grainger Foundation competition is to encourage the U.S. engineering community to become engaged in finding a solution to this specific challenge.The successful technology should address the potable water needs of a rural community of approximately 1,000 residents (roughly 200\u2013300 families). Daily per capita potable water demand is assumed to be 7.5 liters as recommended by the WHO. Such a community might be served by a community-level system, with water piped to convenient distribution points or into homes, or by hand-operated tubewell pumps spread throughout the community. Arsenic can be removed and controlled either at a central community plant or at the household level.In typical villages, women and children usually fetch water. Therefore a treatment process will depend largely on women\u2019s and children\u2019s ability to use the system easily and conveniently. However, community-level systems could be run by a trained operator. Contestants may submit proposals for either type of system.As the reference standard of contamination, specially formulated test water will be used with an arsenic concentration of 300 ppb. This level of contamination is considered to be in the midrange in severely affected areas. The system must reduce this concentration to 10 ppb\u2014presently considered the most acceptable standard by the U.S. Environmental Protection Agency and the WHO\u2014without causing deterioration in other water quality characteristics such as taste and odor, or resulting in an increase in fecal coliforms or other contaminants. The exact formulation of the test water and the complete testing protocol will be made available by the NAE. Contestants may assume that electricity is available for community-level systems because rural areas in many developing countries do have electric power. However, a point-of-use system that does not require electricity will also be considered. If electricity is required, the full life-cycle cost of providing it in all locations must be factored into the economic feasibility estimate.On a per-person basis, the winner will maximize the volume per unit time while minimizing the cost and arsenic content. Because 10 ppb is the WHO standard, no extra credit will be given for attaining lower concentrations. Obviously, this metric allows for some tradeoffs. However, the other constraints described in the Technical Performance section above must not be compromised.All system components should be capable of being manufactured and serviced locally at the village level. Contestants must develop a business plan to demonstrate how this might be done in a specific rural environment and must demonstrate the potential for scale-up to thousands of units. Contestant must also have a plan for disseminating the technology through a nongovernmental organization, the private market, or government, and for covering all costs. All technology developed by the contestants will remain their property. The NAE will hold all information conveyed to it by the contestants in confidence, except that the winning contestant must agree to disclose the winning technology at the time of announcement of the award in February 2007. It is the sole responsibility of all contestants to apply for such patent protection as they deem necessary.Social acceptability must be demonstrated either through carefully documented field experience and/or by drawing upon existing research in social/behavioral science relevant to the challenge. Each contestant must submit a credible field monitoring plan describing how well the system is expected to work, and over what period of time and at what level of maintenance. The plan should also pinpoint expected operational problems and explain how they might be handled.Entries will be judged by a panel of NAE members appointed by the NAE Council. Panel members will be prominent members of the engineering, scientific, industrial, government, educational, and environmental communities. The panel will base its selection of a winner on the criteria outlined in the three requirements sections above with the assistance of specialists familiar with the arsenic problem.http://www.nae.edu/nae/grainger.nsf/weblinks/NAEW-68HUZT?OpenDocument.Under the conditions of the NAE/Grainger Foundation Prize Agreement, the Grainger Prize is available only to a U.S. citizen or team of U.S. citizens, who must be living at the time of notification of selection as prize recipients in the United States. All team members must agree and stipulate in writing in advance how they wish the prize money to be distributed. Such stipulation, once made, shall be irrevocable and unchangeable. The NAE accepts no responsibility for adjudicating disputes of any kind among team members. All expenses incurred by the contestants in pursuit of the prize are their responsibility. For the most up-to-date announcement, see jfritz@nae.edu.Contact: Jack Fritz, NAE, 500 Fifth Street NW, Washington DC 20001 USA, 202-334-2491, e-mail:As part of its global health initiative, the John E. Fogarty International Center (FIC) of the National Institutes of Health (NIH), in partnership with the National Eye Institute (NEI), the National Heart, Lung, and Blood Institute (NHLBI), the National Institute on Drug Abuse (NIDA), the NIEHS, the National Institute of General Medical Sciences (NIGMS), the Office of Behavioral and Social Sciences Research (OBSSR), the Office of Dietary Supplements (ODS), and the Office of Research on Women\u2019s Health (ORWH), invites applications from current and former NIH-supported foreign research trainees to compete for funds that will support their research efforts upon return to their home countries.As junior scientists complete training programs in the United States, many find it difficult to secure the support needed to continue their research projects and careers in their home countries. This Global Research Initiative Program (GRIP) provides the opportunity for junior foreign scientists to compete for such funds through a peer-reviewed process. This is a critical adjunct in the continuation of promising independent research careers that will be of benefit to the investigators\u2019 home countries and the world at large. Women and underrepresented minority scientists in their countries are especially encouraged to apply for these reentry grants. Project proposals should be geared toward the research interests of the applicant and focus on high-priority health and health care problems in the investigator\u2019s home country that also carry global importance, and are of interest to the collaborating institutes, centers, and offices.http://www.drugabuse.gov/International/HHHRF.html); 4) at least two years of research training experience through the NIH intramural Visiting Fellows Program; 5) one year of training through an f5 international fellowship program and one subsequent year of mentored research; 6) be a recipient of a Long-Term Fellowship award through the Human Frontier Science Program, who comes from a low- or middle-income country, and who has spent at least two years in research training; or 7) at least one year of training in the United States and one additional year of significantly mentored research, in the United States or abroad, leading to a completed master\u2019s degree or doctoral degree, at least partially funded through an FIC research training program, with preapproval by the program director.In order to be eligible, foreign scientists must meet at least one of the following criteria: 1) at least two years of research training experience under an FIC-supported training grant; 2) one year of such training experience coupled with one year of significant, well-documented mentored research experience; 3) one year of the NIDA INVEST Fellowship plus at least one additional year of mentored research development of laboratory capabilities or research projects; 2) training of other potential researchers; 3) publications in local as well as international peer-reviewed journals; 4) participation in workshops, seminars, and international conferences; 5) collaborations with past mentors, as well as with other researchers; and 6) attraction of funding from other sources.This funding opportunity will use the R01 award mechanism. An applicant can request up to two modules of $25,000 each or total direct costs of $50,000 per year, plus facilities and administrative costs to a maximum of 8% for a foreign institution. Applications may have a project period of no less than three years and no more than five years. Because an investigator can receive a maximum of five years of support under the GRIP program, and this specific GRIP award is not renewable, any future application will be considered to be an unsolicited competing application based on this project and will compete with all investigator-initiated applications submitted to NIH through the Center for Scientific Review.http://www.dnb.com/us/. For further assistance contact GrantsInfo by calling 301-435-0714 (telecommunications for the hearing impaired: TTY 301-451-0088) or by e-mail:GrantsInfo@nih.gov.Applications must be prepared using the PHS 398 research grant application instructions and forms. Applications must have a Dun and Bradstreet (D&B) Data Universal Numbering System number as the universal identifier when applying for federal grants or cooperative agreements. The D&B number can be obtained by calling -866-705-5711 or online at The letters of intent receipt dates for this PAR are 22 August 2005; 21 August 2006, and 21 August 2007, with the application receipt dates 21 September 2005; 21 September 2006; and 21 September 2007. The earliest anticipated start date for these awards is July of the year following the receipt date.primacka@mail.nih.gov; Chyren Hunter, Retinal Neurosciences and Oculomotor Systems Program, Division of Extramural Research, NEI, 5635 Fishers Ln, MSC 9300, Ste 1300, Bethesda, MD 20892-9300 USA, 301-451-2020, fax: 301-402-0528, e-mail:clh@nei.nih.gov; Ruth Johnsson Hegyeli, Office of the Director, NHLBI, NIH, 31 Center Dr, Rm 4A07, Bethesda, MD 20892-2490 USA, 301-496-5375, fax: 301-496-2734, e-mail:hegyelir@nih.gov; Steven Gust, International Programs, NIDA, 6001 Executive Blvd, Rm 5-274, Bethesda, MD 20892-9581 USA, 301-443-6480, fax: 301-443-9127, e-mail:ipdirector@nida.nih.gov; Dennis Lang, Division of Extramural Research and Training, NIEHS, PO Box 12233, MD EC-20, Research Triangle Park, NC 27709 USA, 919-541-7729, fax: 919-541-2583, e-mail:lang4@mail.nih.gov; Ann A. Hagan, NIGMS, 45 Center Dr, Rm 2AN24H, Bethesda, MD 20892-6200 USA, 301-594-4499, fax: 301-480-1852, e-mail:hagana@nigms.nih.gov; Virginia Cain, Office of the Director, OBSSR, Bldg 1, Rm 256, 1 Center Dr, Bethesda, MD 20892 USA, 301-402-1146, fax: 301-402-1150, e-mail:virginia_cain@nih.gov; Mary Frances Picciano, ODS, 6100 Executive Blvd, Ste 3B01, Bethesda, MD 20892-7517 USA, 301-435-3608, fax: 301-480-1845, e-mail:piccianm@od.nih.gov; Lisa Begg, Office of the Director, ORWH, 1 Center Dr, Rm 201, Bethesda, MD 20892 USA, 301-402-1770, fax: 301-402-1798, e-mail:beggl@mail.nih.gov. Reference: PAR-05-082Contact: Aron Primack, Division of International Training and Research, FIC, Bldg 31, Rm B2C39, 31 Center Dr, MSC 2220, Bethesda, MD 20892-2220 USA, 301-496-4596, fax: 301-402-0779, e-mail:"} +{"text": "Using the Internet for laboratory needs to quickly retrieve information, such as special parameters, addresses, market news & innovations or a product's manual, is time-consuming. There exist already numerous Web sites and their number is ever-increasing. But does the useful information presented in these sites always reach the desired target group? Evidence shows that this has not happened, so far. IVD-net has identified this problem and devoted a good deal of effort to resolving it, over the last year. As an alternative to time-consuming searches in a large number of sources, IVD-net offers a centralized, firmly established Internet forum devoted to in vitro diagnostics.IVD-net intends to be a guiding compass in the \"jungle\" of Web-pages dealing with laboratory medicine. This guidance is being created in a virtual community of specialists and interested people this field. It is the community of people connected by a common interest in in-vitro diagnostics. In this community, step-by-step, a network of knowledge and experiences is created, where each member can benefit from. IVD-net is authentic; there is no contribution without reference to the original source.Structure of IVD-Web-sites:German and English languages.\"public\" pages for welcome and orientation.interactive \"members'\" pages for registered members.Membership is free of charge. Members pages consist of:News: daily new information and scientific events.News Group: the bulletin board for your questions, answers, comments, information.Addresses : list of addresses and/or home pages of important partners in laboratory medicine directly linked to their URL.Archives: thematically arranged & handy for research.Hardware and Software: marketplace of instrumentation and a software catalogue. This is a virtual catalogue with efficient search functions, direct ordering options and attractive special offers of alternative suppliers!Economic globalization and increasing pressure to reduce costs in public health systems bring up a great challenge; the one of finding new solutions."} +{"text": "Human pluripotent stem cells (PSCs) offer a scalable alternative to primary and transformed human tissue. PSCs include human embryonic stem cells, derived from the inner cell mass of blastocysts unsuitable for human implantation; and induced PSCs, generated by the reprogramming of somatic cells. Both cell types display the ability to self-renew and retain pluripotency, promising an unlimited supply of human somatic cells for biomedical application. A distinct advantage of using PSCs is the ability to select for genetic background, promising personalized modelling of human biology \u2018in a dish\u2019 or immune-matched cell-based therapies for the clinic. This special issue will guide the reader through stem cell self-renewal, pluripotency and differentiation. The first articles focus on improving cell fidelity, understanding the innate immune system and the importance of materials chemistry, biofabrication and bioengineering. These are followed by articles that focus on industrial application, commercialization and label-free assessment of tissue formation. The special issue concludes with an article discussing human liver cell-based therapies past, present and future.This article is part of the theme issue \u2018Designer human tissue: coming to a lab near you\u2019. These range from coordinated tissue morphogenesis during gestation, to tissue renewal and homeostasis in the adult. Essential to these processes is hierarchical control of cell potency. In the developing embryo and the adult, cell fate is determined by niche-specific factors and executed through defined changes in gene expression. A good example of cell fate determination is observed in PSCs, with the stem cell master regulators, Oct-4, Sox 2 and Nanog serving to instruct stem cell self-renewal and differentiation . During development iew, see ).totipotent blastomere formation, capable of forming embryonic and extra-embryonic tissues. At the point of blastocyst formation, the pluripotent cells of the inner cell mass are capable forming all three germ layers in the developing embryo, with the trophectoderm contributing to extra-embryonic tissues [multi, bi and unipotent stem cell populations persist in the neonate and the adult, serving to instruct development and/or tissue maintenance.Development starts with the fertilized egg cell, and tissues . As devein vivo a longer-term aspiration. To date, a number of hESC-derived products are in clinical trials, including macular degeneration, diabetes and heart disease, with some other applications registered for clinical trial approval [Since the 1980s, PSCs have taken centre stage as a promising cell candidate to model and treat human diseases. The successful isolation and culture of murine PSCs was presented in 1981 and heralded a new era in cell biology, driving important advances in our understanding of mammalian physiology ,4. It waapproval \u20138. iPSCsThe landmark discovery that mammalian DNA, from a fully differentiated cell, could be reprogrammed to pluripotency leading to the birth of viable offspring inspiredSince those seminal studies, there has been a global push to exploit PSC-based technology to challenge our understanding of human biology and to treat disease. Key outputs from these studies have been the generation of high-fidelity human models \u2018in a dish\u2019 and pioneering cell-based therapies to treat human disease. Stem cell-derived models have been developed for many tissues, including gut, liver and brain to name a few \u201316. Stemin vitro is the recreation of the nascent tissue structure, with the appropriate niche factors [in vitro and in vivo have seen scientists from chemistry, biology, physics and engineering backgrounds working together. This has led to exciting developments in human tissue engineering. With this in mind, we have prepared this special edition of the journal and invited contributions from experts around the world to describe cutting-edge activity in their fields and future directions.Annually, more than 115 million animals are used worldwide in experimentation . While t factors . Attemptet al. [et al. [et al. [We begin with a review of the progress made in the field of PSC reprogramming and self-renewal written by Abu-Dawud et al. . This iset al. , focuses [et al. . The nex [et al. , reviews [et al. . This is [et al. . Followi [et al. present in vitro engineered tissue. Underpinning success in technology scale-up and application is the development of the raw materials required to produce the somatic cells at clinical grade. A key consideration is the extra-cellular matrix used in the production process and this is reviewed by Hagbard and co-workers [Essential to the generation of cell-based models and therapies is the quality of the -workers . Recentl-workers ,32. Such-workers . This is-workers dealing in vitro [In recent years, tremendous progress has been made in the development of organoids from human tissue ,36. Thesin vitro . Recent in vitro . Followiin vitro discuss in vitro . Stem cein vitro . For thein vitro .The ability to produce high-fidelity human tissue from stem cells for basic and clinical application requires the development of non-invasive monitoring technology that can measure and report in real time. The merits of label-free monitoring are reviewed by Gamal and co-workers . We end We are extremely grateful to the authors, the reviewers and the editorial team at Philosophical Transactions B for their time and effort in delivering this special edition of the journal. We hope that this collection of papers stimulates interest and collaboration within the field, with a focus on translating basic scientific concepts into game-changing regenerative medicines for the clinic."} +{"text": "There is an error in the last sentence of the Acknowledgments section. Specifically, the research group number is incorrect. Please see the correct sentence here: The authors would like to extend their sincere appreciation to the Deanship of Scientific Research at King Saud University for its funding this Research Group No. RG-1435-025.There is an error in the Funding Disclosure statement. The correct Funding Disclosure statement should read: Funding was provided by the Department of biotechnology, Government of India as project BT/PR 13194/BRP/10/742/2009 and DST SR/SO/BB-0018/2011 to RHK; the University Grant Commission, New Delhi, MANF-SRF to SN and MZ; the University Grant Commission, New Delhi, UGC-post doctoral fellowship to NZ; the University Grant Commission, New Delhi, UGC-SRF (NET) fellowship to MRA; the Department of Biotechnology (DBT) DBT-JRF (NET) fellowship to MKS; and the Deanship of Scientific Research at King Saud University for its funding this Research Group No. RG-1435-025. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Dr. Lian-Cang Xu \u5f90\u8054\u4ed3, 1927\u20132015), born in Haining County, Zhejiang Province, China, is a well-known Chinese psychologist Pedagogical Science AcademyScience China, received much attention from Western scholars. Besides that, he focused on the stimulus-response compatibility and the mediating effect of language in human-computer interaction. In the late 1960s, Dr. Xu participated in the development project of Chinese first man-made satellite \u201cDongfanghong-I\u201d. As a member of the leaders, he and his group members finished the first observation experiment of the animals under the weightlessness condition in China. Furthermore, he revealed the physiological and psychological changes of the experimental animals under such conditions\u00a0 in 1970s. To rehabilitate the IPCAS and Chinese Psychological Society (CPS) in a short time, Dr. Lian-Cang Xu tried his best to take a lot of measures with other colleagues. In such difficult environment, he still completed two projects: one was the biological effect of laser, specifically on vision, and the other was the earthquake forecasting through the observation of animals\u2019 reaction. As soon as IPCAS was formally reinstated in 1976, he went back to study management psychology and led a team to carry out a survey on employees\u2019 job and life satisfaction, which was appreciated as \u201cChinese Gallup Poll\u201d by New York Times.Since 1979, Dr. Lian-Cang Xu began to lay stress on the action research which centers on the organization development. A tool for measuring the effectiveness of concrete managing policies was developed through his efforts, and its validity was verified in oil, coal, aviation, and railway industries. According to the data obtained by this measuring tool, he discovered the causal relationship between economic performance and human resource management. Moreover, he and his partners conducted studies on leadership behavior, management decision-making, personnel training, and risk awareness, all of which were pioneering in China, and he also integrated the productivity, relations of production and culture to propose a \u201cTripility Theory\u201d, triggering much attention from academia of China and other countries. Cooperating with other world-famous scientists, such as Misumi Juuji, Hofstede, and Federer, he did a series of cross-cultural researches and devoted to finding out whether Management Theory has cultural specificity and applicability in different countries. All these research findings made significant contribution to the discipline construction of management psychology and promoted the development of management science in China.In order to cater for the needs of economic system reform and promote the research and application of behavioral science, Dr. Lian-Cang Xu and some other scholars appealed for the establishment of Chinese Behavioral Science Society in 1980s. Meanwhile, he paid more attention to the research of management, such as staff training, work values analysis, and other aspects. Apart from the indigenous study, he began to focus on the research progress in other countries, for example, the PM (Performance Maintain) method from Japan and the Repertory Grid method from U.S. The introduction of these methods to China enhanced the development of industrial psychology research effectively.Management Psychology (1986) with Sheng-Zhong Lu (\u5362\u76db\u5fe0), Organizational Management Psychology (1988) with Wen-Quan Ling (\u51cc\u6587\u8f81), Research on Leadership Behavior (1991) with Lin-Lin Yang (\u6768\u6797\u6797), Organizational Behavior (1993), Management Psychology and Its Application: Serve for the Management in Hospital (1993), and Management Psychology out of Jungle (2007), and more than one hundred papers on international and Chinese journals : one second class prize for STP of Chinese Academy of Sciences (CAS), one second class prize for STP of National Light Ministry of ChinaDr. Lian-Cang Xu had abundant works through his life, including more than ten academic monographs, such as s Liang, . He and avior 199, ManagemActa Psychologica Sinica, etc\u00a0, secretary general of CPS (1978\u20131985), executive committee member of International Association of Applied Psychology , vice-president of Chinese Behavioral Science Society (1985\u20131988), vice-president of Chinese Association of Social Psychology (1990\u20131993), and the chief editor of etc\u00a0Jin, .As a model in Chinese psychological circle, Dr. Lian-Cang Xu studied rigorously, explored diligently, and dared to innovate. For the development of Chinese psychology, he was strongly willing to be dedicated to his work, and trained a group of psychology doctorate and master for China. His conscientious working attitude and generous character were deeply respected all over the world."} +{"text": "Zoological Research (ZR). In recognition of its impressive progress and great potential to develop into a leading journal in the field, ZR is now supported by the \"Project for Enhancing International Impact of China STM Journals\" (PIIJ) (Class B) (2016-2018). Having wide coverage, PIIJ is the most influential project in China for supporting and promoting the development of English language academic journal publications. In addition, ZR has achieved the \"International Impact Academic Journal of China\" title five years in a row. The year 2016 was a wonderful and important one for ZR depends on its unique niche in the field and the joint effort of its strong editorial team. Emphasizing the scope of ZR, namely, \"Primates and Animal Models\", \"Conservation & Utilization of Animal Resources\", and \"Animal Diversity and Evolution\", two special issues were released in 2016: \"The Amphibian and Reptile Biodiversity of Qinghai-Tibetan Plateau\" and \"CRISPR/Cas9, Model Animals and Human Diseases\". Moreover, to promote communication and collaboration among peer researchers, ZR is hosting the \"2017 Frontiers in Zoology Symposium\" in February 2017. The success of ZR continues to recruit local and overseas scientists to serve on its board. In 2016, ZR engaged four renowned scientists with varied expertise to join the vibrant ZR editorial team. These new board members include: Dr. Wai-Yee Chan, Chair Professor and Director, School of Biomedical Sciences, Chinese Hong Kong University, Hong Kong SAR, China; Dr. LeAnn Blomberg, Beltsville Agricultural Research Center (BARC), United States Department of Agriculture, USA; Dr. Hua-Xin (Larry) Liao, Professor, College of Life Science and Technology, Jinan University, China, and Professor, Duke University, USA; and Dr. Meng-Ji Lu, Professor, Institute of Virology, University of Duisburg-Essen, Germany. In addition, Dr. Guojie Zhang, a well-known expert on animal genomics who holds a position at the China National GeneBank, BGI-Shenzhen, and the Department of Biology, University of Copenhagen, as well as a guest professorship at the Kunming Institute of Zoology, Chinese Academy of Sciences, will also join the editorial board. With these new members, the editorial board of ZR now includes 48 distinguished scientists with expertise across multiple disciplines. To further strengthen the editorial board and widen its scope of expertise, ZR has continued to promote and market the journal during this past year. A new and more interactive homepage was launched and, as an open access journal, full-text articles can be downloaded via the homepage, PubMed Central, and SciEngine. To expand our readership, abstracts of published articles are also available online in Chinese. Information regarding the new initiatives and measures of ZR will continue to be shared with our readers through our official WeChat account (ZoolRes). In addition, ZR has now been indexed by Scopus and other related databases, and has a CitesScore value of 0. 48 and ranks 786 among 1 549 journals in the subject \"Medicine\". To further its scientific reach, ZR will continue to publish innovative zoological studies and advance knowledge in scientific research. With the proud tradition of attracting outstanding scholars with diverse backgrounds and expertise to write for the journal, as well as the hard work and dedication of our editors and editorial board members, ZR has become a representative international scientific journal in animal studies. With a growing number of international submissions and article citations, as well as an increasing impact factor, we believe that ZR will confidently march into its thirty-seventh year and accomplish its goal of becoming an iconic journal in animal research. ZR could not have achieved what it has without the support of you, our readers. It is a journal that belongs to all members of the animal research community, and we look forward to continuing to work with you all to nurture and carry ZR to the next stage in 2017. SincerelySchool of Biomedical Sciences, The Chinese University of Hong Kong, ChinaWai-Yee Chan, Executive Editor-in-Chief Kunming Institute of Zoology, ChineseAcademy of Sciences, Kunming 650223, ChinaYong-Gang Yao, Editor-in-Chief"} +{"text": "Scientific Reports7: Article number: 4328510.1038/srep43285; published online: 03072017; updated: 05182018The Acknowledgements section in this Article is incomplete.\u201cWe thank Seungkyu Ha, Yera Ye. Ussembayev, Richard Janissen, and Hubertus J. E. Beaumont for discussions, Richard M. Berry and Ren Lim for providing the strains, and Theo van Laar for the remaining help with the bacteria. This work is supported by NanoNextNL, a micro and nanotechnology consortium of the Government of the Netherlands and 130 partners, and by the Foundation for Fundamental Research on Matter (FOM).\u201dshould read:\u201cWe thank Seungkyu Ha, Yera Ye. Ussembayev, Richard Janissen, and Hubertus J. E. Beaumont for discussions, Richard M. Berry and Ren Lim for providing the strains, and Theo van Laar for the remaining help with the bacteria. FP was supported by the European Research Council under the European Union\u2019s Seventh Framework Program (FP/2007\u20132013)/ERC Grant 306475. This work is supported by NanoNextNL, a micro and nanotechnology consortium of the Government of the Netherlands and 130 partners, and by the Foundation for Fundamental Research on Matter (FOM).\u201d"} +{"text": "We are delighted to present within this meeting report the abstracts of the \u201cBIT\u2019s 1st World Congress of Biomedical Engineering 2017\u201d which has been hold in Xi\u2019an in China .We were very glad that the congress could take place in the exciting, traditional city Xi\u2019an in China. Xi\u2019an is the capital of Shaanxi Province. It is one of the oldest cities in China, and Xi\u2019an is the oldest of the four great ancient capitals, having held the position under several of the most important dynasties in Chinese history. Xi\u2019an is the starting point of the Silk Road and home to the Terracotta Army of Emperor Qin.This report presents abstracts from an impressive international speaker panel. In total, more than 50 speakers from countries from all over the world like Australia, Austria, Brazil, Canada, China and regions of China, Colombia, Czech Republic, Denmark, France, Georgia, Germany, Hungary, Iceland, India, Italy, Japan, Mexico, New Zealand, Peru, Poland, Qatar, Romania, Russia, Slovakia, Korea, Spain, Switzerland, Republic of Kazakhstan, Turkey, and the United States have presented their latest research results. The international term \u201cbiomedical engineering\u201d is understood to mean the interaction between medicine and technology for the development of equipment for diagnosis, therapy, and prevention of diseases. Physicians and engineers are equally involved in solving these problems. However, solutions are only possible if the pathological situation or the interaction of all relevant factors is known, so that starting points are identified at which the engineer is able to aid medicine.Technology has already entered many areas of medicine, as a comprehensive scientific literature review confirms. Just think of X-ray and irradiation devices, measuring and monitoring instruments of various kinds as well as surgery and prosthesis technology, and many others. However, these are not the only research areas addressed in the present congress. In addition, areas are selected that are difficult to access in a technical solution. An attempt is made to interpret the language of the physician in such a way that the technical and scientific core of the problem becomes apparent to the engineer. In the majority of the lectures, therefore, the medical problem is presented, after which the technical aspects of the equipment or research are discussed.Keynote ForumNeural EngineeringBiomedical Imaging and BiosensorsInterventional Technology and TreatmentDigital Medicine and Big DataTissue Repair and Regenerative MedicineCell EngineeringNanomedicine and EngineeringPhysical Therapy and RehabilitationFor a better overview, the entire area of biomedical engineering was divided into the following sessions:Medicines, I also want to inform you that we have started a Special Issue named \u2018New Innovations in Biomedical Engineering\u2019 and that all the readers of this report are cordially invited to submit original papers or short reports especially regarding the lectures of this congress for publication in this Special Issue.As editor-in-chief of Chen, J.-H.AbstractCone-beam computed tomography (CBCT) systems have been developed to provide anatomic images of patients in order to enhance the accuracy of diagnosis. In the reconstruction process, accurate geometry is extremely critical to obtain high quality reconstructed tomographic slices. Especially in high resolution CBCT, a bit of misalignment in geometry can seriously degrade the quality of the reconstructed images. There are several geometric calibration methods developed which use a dedicated phantom to acquire geometric parameters. The projections with geometric misalignment can be calibrated by re-mapping the projections with the estimated geometric parameters. Then, the observed object can be reconstructed with the calibrated projections. In this study, a new phantom which consists of 26 steel balls precisely located in a cylindrical-shaped structure is applied to calibrate the geometry of a preclinical CBCT system. There are two circular patterns in the cylindrical phantom which consists of 12 steel balls spaced evenly over 360 degrees in each circle. The remaining two steel balls are located at the center of the two sides of the cylindrical phantom. The calibration process with the proposed phantom is described in the following. First, the projections of the two circular steel ball patterns were utilized to fit two ellipses by least squares. Second, the converging points of the two ellipses were calculated by the elliptical functions. Third, the magnifying factor is estimated by the other two steel balls. Finally, the complete geometric parameters including three rotation parameters of the detector, the location of the piercing point and the source point position can be obtained by solving the analytical equations. After that, the geometric parameters are applied to re-mapping the projection data. In order to validate the proposed method, a prototype preclinical CBCT gantry is applied. The proposed calibration phantom and the hydroxyl-apatite (HA) phantom were scanned by the real gantry system. After geometric calibration by re-mapping the projection data with the geometric parameters, the HA phantom images were reconstructed by the filtered back projection (FBP) algorithm with the calibrated projections. Comparing before and after calibrated reconstructed slices of the HA phantom, the artifacts caused by misalignment of geometry are obviously reduced.BiographyDr. Jyh-Cheng Chen received the B.S. degree in physics from National Central University, Taiwan, in 1983, and the M.S. degree in physics and Ph.D. degree in optical sciences from the University of Arizona, USA in 1988 and 1995, respectively. In 1995, he joined the Opto-Electronics and System Laboratories, Industrial Technology Research Institute, Taiwan, as a Research Associate working on semiconductor laser packaging. In 1996, he became a member of the faculty of Division of Radiological Science and Technology, Department of Medical Technology, National Yang-Ming University (NYMU), Taiwan. In 1998, he became an associate professor of Institute of Radiological Sciences, NYMU, Taiwan. Since 2005, he has been a Professor in the Department of Biomedical Imaging and Radiological Sciences (BIRS), NYMU, Taiwan.Cho, K.AbstractIn Korea, PHR is being used to make a nationwide approach to improve the health of each individual.Cancer, cerebrovascular disease, diabetes, etc., and provides a method of disease prevention for each individual. In the case of stroke, a 10-year stroke prediction model was developed and categorized as a stroke using the Korean national health examination data. Then, we will develop the algorithm to provide a personalized warning on the basis of each level of stroke risk and a lifestyle correction message about the stroke risk factors.The AUC values of stroke risk prediction models, developed using multiple risk factors, were 0.83 in men and 0.82 in women, which showed a high predictive power. The probability of stroke within 10 years for men in the normal group (less than 50 percent) was less than 3.92 percent and in the very risky group (top 0.01 percent) was 66.2 percent and over. The women\u2019s probability of stroke within 10 years was less than 3.77 percent in the normal group (less than 50 percent) and 55.24 percent and over in the very risky group.A lifestyle correction message by each risk factor was developed and set up automatically in the individual PHR.BiographyProfessor Cho is currently the President of APAMI , Chair of Health informatics committee, NHIMC Ilsan hospital, Professor of Yonsei medical colleges. He majored in M.D and Family Medicine at Yonsei University College of Medicine, Master of health policy at the Graduate School of Public Health, Ph.D. at the graduate school, and Geriatric fellowship at the University of Rochester, USA for two years. Many projects have been carried out in the field of family medicine, geriatrics, health policy, quality improvement, and medical informatics, with over 40 of papers and book publications. Since 2009, he has been in charge of the national health information exchange project DCM project PI, for three years from 2012, the head of NHIMC Hospital EMR, from 2014 onwards, he is currently in charge of the National Disease Prediction Project. Currently, he is a member of National Medical Information Standardization Committee and DCM committee chair.Litscher, G.AbstractThe application of near-infrared spectroscopy (NIRS) in different fields of research has gained importance, and the number of scientific studies has grown extensively. Within the current keynote lecture, a retrospect and highlights from 1995 to 2017 from the interdisciplinary NIRS research team at the Medical University of Graz, Austria, Europe will be presented. The main topics are results from transcranial cerebral oximetry in the hyperbaric environment and the non-invasive assessment of oxygen metabolism using NIRS technology during high altitude trekking in the Nepal Himalayas. In addition, NIRS publications from intensive care medicine and laser acupuncture are shown. Last but not least, the application of regional oxygen saturation in regenerative medicine is demonstrated. NIRS could be a fruitful approach helping to explore deeper biological mysteries within the brain and the periphery. Special results from laser medicine will also be presented briefly.BiographyProf. Gerhard Litscher is a biomedical engineer and the head of the Research Unit for Complementary and Integrative Laser Medicine, of the Research Unit of Biomedical Engineering in Anesthesia and Intensive Care Medicine, and of the TCM Research Center Graz at the Medical University of Graz, Doctor of technical and Doctor of medical sciences, international lectures, about 600 scientific publications (>190 of them SCI/PubMed-listed), partly on biomedical engineering and on basic acupuncture research, author and/or editor of 13 books, currently editor-in-chief and/or member of the editorial board of more than 35 international scientific journals , Medicines, Integrative Medicine International, Integrative and Complementary Medicine; Associate Editor for Medical Acupuncture, Associate Editor of the Journal of Acupuncture and Meridian Studies (JAMS), one of the editors and lead guest editor of Evidence-based Complementary and Alternative Medicine (eCAM), and Associate Editor of BMC Complementary and Alternative Medicine). Prof. Litscher\u2019s special interests are computer-based High-Tech Acupuncture Research and Neuromonitoring. He is University Professor at the Medical University of Graz and Honorary/Guest Professor at eleven institutions and universities in Asia.Caesar, I.AbstractThe same gene is expressed in functional individuals and species by metacentric chromosome, and in dysfunctional individuals and species via acrocentric chromosome. Metacentric chromosome is analogous to a well-centered and focused eye crystal, while acrocentric chromosome is analogous to a myopic or farsighted eye. Chromosome is a lens that allows our cells to focus the needed wave information during cell division, analogously to a crystal in our eye. Thus, Biomedical Engineering is possible only via mastering the Geometrical Optics of Chromosomes by Laser Bioholography. Current genetic engineering via cutting and pasting DNA snippets destroys the systematic wholes of chromosomal bioholograms and the refraction of the geometrical optics in chromosomes, and, therefore, causes genetic collapse. The Quantum Leap in the Biomedical Engineering is the cost of our survival. Biomedical Engineering as Bioholography is based on Quantum Physics, and understanding the non-local nature of the Bioholograms. Since universe is holographic, we can record and transmit only holographic information, based upon changing the refraction of the scalar wave diffraction grating in chromosomes. According to Holographic principle, Universe is entirely in its every matrix point. The implication of the Holographic Principle states that if Universe is entirely in its every matrix point, then, every matrix point is not simply different from any other matrix point, but is unique. This means that every wave matrix is both non-local, and unique. Thus, if we have two exact copies of the same non-local wave matrix, we can transmit information instantaneously and remotely, from one copy of the same non-local unique wave matrix to another copy. Thus, the precision in the Biomedical Engineering is achieved not via the precision of localization, but, vice versa, via the precision of refraction in the unique and non-local bioholograms aka \u201cwave crystals\u201d. The secret in the remote Biomedical Engineering as Quantum Biocomputing lies in the recording and instantaneous transmission of a structural phantom. I will introduce the new method for recording and remote transmitting of bioholograms via laser spectroscopy and coding electrets on nanolevel via laser. The technology creates \u201clenses\u201d assisting our chromosomes in correcting their refraction for focusing genetic bioholographic information, both locally and remotely.BiographyIrene Caesar, Ph.D., is a Founder of Matrix City LLC, Russia, company creating global subscription platforms for bioelectronic drugs (founded 2016); Founder and President of Wave Genome LLC, company producing bioelectronic devices and software for the distribution of bioelectronic drugs for rejuvenation only (founded 2010); Co-founder of \u201cMatrix City\u201d Consortium with the Institute for National Security in Moscow for building self-sufficient human settlements based upon the remote management of biosystems, climate and geophysical processes for the first time in the history of humankind (founded 2012), presented in the Honorary Lecture at the Harriman Institute of the Columbia University in September 2012; Co-founder of the Quantum Biointernet for the remote rejuvenation via distant laser signal, commercially offered for the first time in the history of humankind by Irene Caesar\u2019s company Wave Genome in May 2013; Colonel of Irkutsk Cossack Military, awarded the Medal of Faith and Service to Russia (2014). Dr. Caesar received her doctoral degree from the Graduate Center of the City University in New York.Mei, L.Abstractext was negative, related to being at the first node of the activating function virtual anode and if its absolute value was larger than that generated by the transmembrane potential gradient, iaxo. In high-frequency stimulation (>4\u20135 kHz), the M-gate at the first node of the virtual anode is locked in the closed position, while the H- and N-gates are locked open. Therefore, the ion current iion is small and can be ignored. This blocking phenomenon only occurs in large diameter fibers in response to a monophasic stimulus.High-frequency stimulation of dorsal column axons and their potential mechanisms for achieving pain relief was explored. A threshold charge rate curve was developed to locate a critical axonal node in an axon to determine where and when action potentials (APs) may be generated. Blocking of the axon\u2019s ability to generate APs occurred if an equivalent current generated by the field potential gradient iBiographyMr. Longzhi Mei is senior research assistant of Department Neurosurgery, Beth Israel Deaconess Medical Center, Boston, MA, USA. He is interested in Neural Simulation and has published 6 papers in electrode stimulation and mechanisms of action in neuromodulation devices. He received his B.S. in Physics at Chengdu University of Science and Technology, China in 1982, and taught general physics in the Southwest Agricultural University 12 years. He then went studied in the United States, receiving an M.A. in Biology at Lehman College, City University of New York, USA 1996. He received his M.S. in Computer Science, Rivier College, New Hampshire, USA.Wen, G.AbstractAmong signals such as psychological signals and forehead biosignals, facial expression is the most effective way for humans to express emotions. This is why it has attracted much attention in machine learning applications in recent years. Typical applications include emotional health, sentiment analysis, and human-computer interaction. Generally, facial expression recognition is performed by machine learning methods. However, their performance are far away from that of human being, as they did not consider significant cognitive laws. For example, for human being, the inertial thinking schemes can be formed through learning, which are then applied to quickly solve the coming similar problems. When problems are heavily different, the inertial thinking generally present the solutions that are definitely imperfect. In such case, Human being will apply the creative thinking such as the reverse thinking to solve problems. Although machine learning methods also form the inertial thinking through learning the rules from a large amount of data. However, when the testing data are of much difference, the formed inertial thinking will inevitably generate errors. This inertial thinking is called as the illusion inertial thinking. Because all machine learning methods do not consider the illusion inertial thinking, it is suggested that the reverse thinking can be considered. It increases the generalization ability of the machine learning methods. The conducted experimental results on benchmark datasets validated the proposed method.BiographyGuihua Wen is a professor of Artificial Intelligence (AI) and head of machine learning and data mining group in South China University of Technology, Guangzhou, China. He majors in emotion recognition, deep learning, large scale data mining, and applications to health care, including Traditional Chinese Medicine. Currently his research are funded by 4 projects from national and local governments. He has published over 30 papers in the international conferences such as famous AAAI and IJCAI, and journals such as Pattern Recognition and Appl. Soft Comput. His research achievement on facial expression recognition was ever reported by Yangcheng Evening News in China.Segura-Saldana, P.AbstractCardiac varices are a very little described entity, many consider it as rare; But our work group considers that it is underdiagnosed, because it is usually asymptomatic, and with routine imaging methods like Chest X-ray or echocardiography are usually imperceptible, this is when Cardiac Magnetic Resonance becomes important, but it is still expensive and an examination usually takes 2 to 3 h, reasons why cardiac magnetic resonance is not done routinely. We have developed an abbreviated protocol in Magnetic Resonance to do these exams in approximately 20 min. In addition, we are working on SDR (SOFTWARE DEFINED RADIO) and algorithms of correction by mistake, we believe that we can optimize the obtained in the conventional magnetic cardiac resonance and with this to improve the diagnosis of cardiac varices.My presentation will be about the problem of diagnosing an entity that is rare, to keep it in mind. And also the treatment and prognosis of this kind of patients.BiographyPedro Segura-Saldana, I am cardiologist at the Rebagliati Martins Hospital , CEO of a Biomedical Company in Peru, research and work focused on solving cardiologic, metabolic and oncological problems from a technological perspective. I am a part-time Professor of Biomedical Engineering in the Dual Degree Program of the Universidad Peruana Cayetano Heredia and of the Pontifical Catholic University of Peru. I have published several articles in journals in my country and also in scientific journals at the United States of North America and Europe, I won Grants for the development of medical devices like arrhythmia detector and the infarct in real time with cellular technology, monitoring the Central Blood Pressure in a non-invasive way, also shortened Magnetic Cardiac Resonance protocols (Grant funded by the UK) among others and I am currently developing intelligent methods in the processing of cardiac magnetic resonance. I am a current member of the Peruvian Society of Cardiology and the Peruvian Society of Hypertension. We are carrying out the National Registry of Cardiac Tumors in Peru, will be the first National registry of cardiac tumors.Xu, S.AbstractTo monitor the temperature distribution of a cell and its changes under varied conditions is an important fundamental issue and currently remains as a technical challenge. A variety of non-contact methods used for measuring cellular temperature have been developed, where changes of local temperatures at cell-level and sub-cell-level are indirectly calculated through the changes in intensity, band-shape, bandwidth, lifetime or polarization anisotropy of the fluorescence spectra recorded from the nano-sized fluorescent materials pre-injected into the target cell. Unfortunately, the optical properties of the fluorescent nano-materials may be affected by complicated intracellular environment, leading to unexpected measurement errors and controversial arguments. We have developed high-performance Pd/Cr micron thin-film thermocouple (TFTC) arrays and double-stabilized system with a stability of \u00b15 mK. The local temperature of TFTCs close to adherent human hepatoblastoma (HepG2) cellswere continuously recorded for days, showing frequent temperature increment of 10\u201360 mK and a maximum up to 200\u2013300 mK. Details of the device fabrication, sample preparation, thermal system stabilization and measurement procedures are published elsewhere. The method has set up solid foundation for realization of real-time and precise 2D mapping of local temperature distribution for a single live cell with our nano-sized TFTC arrays, therefore it may valuable information for cell biology and research on drug effects and clinical therapies.BiographyProfessorXuobtained his Ph.D. degree in 1999 from Department of Physics, National University of Singapore. He worked at the \u201cCenter for Superconducting & Magnetic Materials\u201d, National University of Singapore as a Research Fellow in 1999\u20132001, at Physics Department, Pennsylvania State University as a postdoctoral researcher 2001\u20132003, and then at \u201cCenter for Nanoscale Science\u201d, Pennsylvania State University as a Research Associate in 2003\u20132006. Since April 2006 he has been teaching and doing research at the Department of Electronics, Peking University. Currently Prof. Xu\u2019s group mainly works on the underlying physics of electrical communication in biosystems, including soft-material electromagnetic (EM) waveguides, propagation of EM pulses in axons and among cells, working mechanism of brain, etc. The group also works on time-resolved 2D mapping of temperature at the micro-/nano-scales, e.g., a single cell, 3D micro-bio-probes, and solid thermal sensors for ultrahigh temperatures (>2000 K).Lee, C.-J.Abstract2 LASER system offered precise and focused point cutting energy to the septum. Several efforts were applied to prevent RLN injury in the cases descriptions. We use transnasal esophagoscope and eating assessment tool (EAT-10) for anatomic and functional result evaluation. Much improved symptoms of dysphagia and intact RLN function were observed. Under the assist of rigid laryngoscope and point-cutting CO2 LASER, KJD diverticulotomy could be performed safely with little complication for patients refusing transcervical route.Killian-Jamieson diverticulum (KJD) is a rare cervical esophageal abnormality. Transcervical approach has been the main treatment modality to prevent recurrent laryngeal nerve (RLN) injury. We presented several cases of patients confirmed with KJD and were managed successfully under rigid endoscope. The new technique and idea were described in detail. Under rigid laryngoscope, the septum between the true esophagus lumen and diverticulum can be exposed clearly. A microscope equipped with COBiographyDr. Chia-Jung Lee received an M.D. degree from China Medical University, school of Medicine in Taichung, Taiwan, and completed his clinical training in Shin-Kong Wu-Ho-Su Memorial Hospital in Taiwan and with advanced training in Laryngological Surgery at the University of Washington in Seattle with Professor Albert Merati on 2012. He then came back to Taiwan and establish the lab of voice and swallowing in Shin-Kong Wu-Ho-Su Memorial Hospital on 2013. Dr. Chi-Jung Lee focused his clinical interests at airway disease, dysphagia disease and phonosurgery and helps a lot of patients with laryngeal diseases in Taiwan and Southeast Asia. He was the invited speaker of the 13th Asia-Oceania Otolaryngology Head and Neck Congress on 2015. Dr. Chia-Jung Lee holds a faculty appointment in the Shin-Kong Wu-Ho-Su Memorial Hospital, Department of Otolaryngology-Head and Neck Surgery. He was promoted as the Chief of the Otolaryngologic Endoscopic room since 2014.Heer, R.Abstract3N4) waveguide based Mach-Zehnderinterferometric (MZI) photonic sensing platform operating at a wavelength of 850 nm (TM-mode).Inkjet printing is a versatile method to apply surface modification procedures in a spatially controlled, cost-effective and mass-fabrication compatible manner. Utilizing this technology, we investigate two different approaches for functionalizing label-free optical waveguide based biosensors: (a) surface modification with amine-based functional polymers (biotin-modified polyethylenimine (PEI-B)) employing active ester chemistry and (b) modification with dextran based hydrogel thin films employing photoactive benzophenonecrosslinker moieties. Whereas the modification with PEI-B ensures high receptor density at the surface, the hydrogel films can serve both as a voluminous matrix binding matrix and as a semipermeable separation layer between the sensor surface and the sample. We use the two surface modification strategies both individually and in combination for binding studies towards the detection of the protein inflammation biomarker, C-reactive protein (CRP). For the specific detection of CRP, we compare two kinds of capture molecules, namely biotinylated antibodies and biotinylated CRP-specific DNA based aptamers. Both kinds of capture molecules were immobilized on the PEI-B by means of streptavidin-biotin affinity binding. As transducer, we use an integrated four-channel silicon nitride in 2001, and her Ph.D. in Mechanical Engineering from University of Rome \u201cLa Sapienza\u201d in 2007. During her doctorate, she was visiting scientist at the MIC Department of the Technical University of Denmark. Afterwards, she did her research in several laboratories including Silicon Biosystems S.p.A., Harvard Medical School, Kist Europe and most recently at the Italian Institute of Technology. In 2016 she became Associate Professor at the NPU at the Department of Mechanical Engineering. Dr. Simone\u2019s scientific interests include Microfluidics and miniaturization, Lab on a Chip, Microfabrication, Soft Matter and Biophysics. Dr. Simone has published more than fifty of papers, patents and books.Singh, A.AbstractTumor response evaluation in diagnostic imaging has evolved over the past 40 years. The World Health Organization (WHO) response criterion was first introduced in 1979 without any specific imaging stipulations or protocols. Therefore, different groups subsequently proposed modifications and to unify and standardize the criteria, the Response Evaluation Criteria In Solid Tumors (RECIST) was introduced in 2000. However, practical use of RECIST together with the rapid development of imaging techniques and newer chemotherapeutic agents has highlighted the limitations of RECIST and the need for updated criteria. Various newer anticancer drugs that vary in their effects to induce necrosis in the tumor content have demonstrated an inherent limitation and unsuitability of RECIST-based tumor evaluation that assesses only lesion size by single dimensional measurements. Volumetry by automated techniques estimates total tumor volumes and includes necrosis volumes; a tumor fraction that is actually considered a response to therapy. We believe that assessing the volume change of merely the non-necrotic content of the tumors or dynamic cross-sectional imaging-based viable tumor tissues between the pretreatment and post treatment time point imaging scans would be a more reliable predictor of therapy response than the existing RECIST criteria. The illustrated examples in this presentation with in-house conducted research for such methods provides evidence-based support for automated extraction of necrotic components from the tumor tissue to derive the non-necrotic/viable tumor volume estimates that may provide a valid and reliable option for its incorporation with the present diagnostic workflow in the future. This presentation will educate on the limitations of currently used response assessment criteria and will focus on a new set of preliminary studies that will highlight the emerging importance of computer-aided volumetry in treatment response assessment in oncology.BiographyFollowing his medical school training, Dr. Anand Singh was selected among the top ten physicians to pursue my MD program from one the best cancer centers in Asia. He is currently a clinical emergency radiologist and research faculty in the department of Radiology at Massachusetts General Hospital (MGH), Harvard Medical School and member of Dana-Farber/Harvard Cancer Center, USA. His research interests have increasingly inclined on development of newer Computer-aided detection and tumor imaging objective methods over span of thirteen years in Harvard. Dr. Singh\u2019s research career at MGH has resulted in over 140 publications in form of original research articles, peer review articles and book chapters. There are over 600 citations of his research across the globe and he has received several awards, including awarded the Radiological Society of North America (RSNA) research trainee awards, three times (2010 and 2012) and RSNA research scholar grant (2009) at an international platform that is considered to be one of the largest medical conferences in the world. His research work has been instrumental in getting grants nearly 2 Million $ for his research. His research work has also led to formation of computer design and up gradation projects in his lab where automatic quantification of viable liver and sarcoma tumor volumes are under advanced development for clinical translation with promising results. Dr. Singh\u2019s work has also led to innovations of multiple clinical imaging protocols for delineation of liver, pancreatic, biliary, and renal vasculature and bile duct anatomy. Dr. Singh has served as a lead moderator of IAEA weeklong education course for CT radiation dose with participants as representatives from 11 IAEA affiliated countries from Europe and Asia. Dr. Singh is actively involved in MGH and Harvard Global health and outreach wherein his efforts span across various continents in collaborations.Stanciu, G.A.AbstractIn our work we present some investigations on different tissues acquired by using several linear and nonlinear optical microscopy techniques working in far field or in near field. For imaging we used a new multimodal system integrating several microscopy techniques which offer the possibility for investigations at micro and nanoscale on the same area. Far field techniques including confocal laser scanning microscopy (CSLM) and second harmonic generation imaging (SHGI) have hundred nanometers resolution and scattering scanning near field optical microscopy(s-SNOM) has a nanometer resolution. Optical images are correlated with surface topography images acquired by atomic force microscopy (AFM). Our work is mainly focused on collagen structures.BiographyDr. George A. Stanciu, is Professor of Physics at University Politehnica of Bucharest. He is head of Center for Microscopy-Microanalysis and Informatiion Procesing founded by him in 2001. He got his Doctor degree (Ph.D.) in Technical Physics in 1981. He received the Professor title in 1994. Starting with 1973 he has been working in the Laser Scanning Microscopy field. In 1977 his team reported the first Digital Laser Scanning Microscope. He have been working for long time in the frame of laser scanning microscopy (instrumentation and applications) are and also in the scanning probe microscopy (instrumentation and applications). Lately he is focused on biological application of different scanning laser microscopy techniques. Prof. George Stanciu is senior member of the IEEE from 1995.Song, W.AbstractPhotoacoustic microscopy (PAM) is capable of measuring the optical absorption properties in the tissues, which is complementary to the sophisticated optical microscopic technologies, such as confocal microscopy, multiphoton microscopy, and optical coherence tomography, based on various imaging contrasts, including optical scattering, fluorescence, and polarization. The optical diffraction-limited lateral resolution down to the micrometer or even submicrometer scale is achieved by utilizing a microscope objective to tightly focus the photoacoustic excitation laser onto biological samples. However, both acoustic detection sensitivity and frequency response is limited by the piezoelectric ultrasonic transducer, which produces low signal-to-noise ratio and poor axial resolution in PAM. Here, we report a novel ultrasonic/photoacoustic detection method based on sandwiched configuration of graphene between prism and water. Photoacoustic initial pressure transients modulate refractive index of the coupling water, which changes the polarization-dependence absorption of the graphene. As a result, the phtoacoustic detection is realized through recording the reflectance perturbation of the probing beam with a balanced photodiode. The graphene-based sensor shows an estimated noise-equivalent-pressure sensitivity of around 550 Pa over an approximately linear pressure response from 11 kPa to 55 kPa. Moreover, it enables much broader ultrasonic bandwidth detection up to about 150 MHz, primarily associated with the highly localized evanescent field. In vivo PAM imaging of three-dimensional microvasculatures is obtained label-freely in mouse ears because of strong optical absorption of inherent hemoglobin. Our results implies the great potentials of graphene-based PAM for biomedical investigation, such as tumor angiogenesis and neurovascular coupling.BiographyDr. Wei Song is working as Associate Professor in Nanophotonics Research Centre, Shenzhen University. In 2014, he obtained his Ph.D. degree in Department of Physics, Harbin Institute of Technology. Afterwards, he started work as Assistant Professor at Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences. His main research interests are biomedical photonics, including photoacoustic imaging, optical coherence tomography (OCT), multiphoton microscopy, and multimodal microscopic technologies by integrating multiple imaging methods. Dr. Song has published over 20 peer-reviewed papers on the development of novel photoacoustic microscopy, OCT, and multimodal microscopy.Sun, R. and Pinchuck, L.Abstract2-C(CH3)2]n- (\u201cxPIB\u201d) began in 2003. Implantable medical devices made from polyisobutylene co-polymers have been implanted in more than a million patients as drug-delivery coatings for coronary stents since 2004 and have been used in the eye as a glaucoma shunt in over 1000 patients since 2008. XPIB is made ultra-pure with no cleaveable groups; thereby eliminating the possibility of forming glistenings or hazing typical of acrylic-based IOLs. Its high refractive index (1.50\u20131.53), rubbery nature (Modulus at 10% strain < 100 g/mm2), scratch resistance and low glass transition temperature (\u221270 \u00b0C) enables insertion into the eye through cannulas as small as 1.5 mm; thereby eliminating suturing of the cornea. Bonded-in UV blockers provide a UV cut-off at <400 nM with light transmission values of 95%. It\u2019s Abbe number at 23 \u00b0C is 49.8. The unique chemistry of xPIB provides distinct advantages over conventional hydrophobic materials in regard to lack of glistening, low insertion profile, compatibility with silicone oils (vitrectomy patients) and clarity. In addition, it can be injection molded or machined to provide high-quality affordable lenses. From our development, we have demonstrated that the IOLs from xPIB showed significant advantages in both optical and mechanical properties compared to the existing leading IOL products in the markets.Xi\u2019an Pillar BioScience Co., Ltd. is an ophthalmology company founded in January, 2015. Pillar\u2019s mission is to develop the world\u2019s first pre-loaded, hydrophobic, glistening-free, micro-insertable intraocular lens (\u201cIOL\u201d) for use as both a commodity and premium IOL to serve global cataract patients. Work on this novel next-generation IOL material made from crosslinked polyisobutylene - was finally established and these proteins were identified as \u201cearly\u201d and \u201clate\u201d acting molecules respectively, in a very crucial developmental event, that basically transforms \u201cpolyps\u201d to full-fledged \u201ccarcinomas\u201d in COLORECTAL tumors. The detection of these genetic and developmental parameters, served as a focal-point and a prominent diagnostic feature, for detection of effects, i.e., gain/loss of other components involved during progression of CRC disease. Coincidentally, the chromosomes on which these genes reside have been found to be dense and rich in SNPs (hot-spots), the details of which were published in a separate report . This work harnessed the potential of Genetics, Developmental Biology and Bio-Informatics tools to solve a long-standing puzzle in pin-pointing some genetic factors that were critically involved in the progression of CRC disease. The report has created enough impact, in terms of authentically suggesting, that it is only when we deploy a combinatorial approach towards certain complicated biological problems, can we successfully unveil the underlying mechanisms in greater details.http://yournewswire.com/breakthrough-scientists-find-way-to-change-cancer-cells-into-healthy-cells/). My talk would shed light on how we could intelligently utilize these efficient tools together, to attack the \u201cBad seeds\u201d in ways to cure the myriad diseases, like Cancer.However, it is now conceived that, at the heart of every tumor lies a rare sub-population of cells (Cancer Stem Cells-CSCs), which give rise to most of the Cancers and are now the targets of investigation. Since no definitive markers or efficient labeling tools are available, this population of cells still remains elusive in both cancer and stem cell biology. Therefore, it would be critical to understand molecular differences between stem cells and cancer cells, which might be helpful in providing novel insights into the mechanism of tumorigenesis as well as potential therapeutic targets, in foreseeable future. We have come a long way in the stem cell advances over time. Very recent breakthroughs include: (a) The tuning and genetic re-programming of stem cells (iPS cells) by a handful of genetic factors and; (b) The transformation of cancerous cells to normal cells by reversing the genetic changes involved and also restricting the awry cancerous cells by using microRNAs at University of Indore, India. She obtained her BSc (Bachelors degree) in Biological Sciences/Chemistry/Physics, MSc (Master\u2019s degree) in Life-Sciences, and Doctoral degree (Ph.D.) at School of Life-Sciences, University Of Indore. She pursued her post-doctoral ventures at Max-Planck Institute for Biophysical Chemistry (FRG), University Of California-Irvine and University of Pittsburgh (USA). Currently, her projects mainly focus on translational-research and extrapolation of basic developmental mechanisms from model-systems like fruitfly (Drosophila) to human. Apart from this, her thrust areas of research interest include; Cancer Biology, Stem-Cell Biology and Homeotic-Gene Regulation. She is keen on studying in details the genetic factors, which presumably aid in understanding of mechanism by which \u201ccancer stem cells\u201d function in transforming a tissue from normal to cancerous states. Her research has a motive to further facilitate the perception of stem cell potential/mechanistic in areas of Regenerative Medicine, Translational Research and Anti-cancer therapy. Being involved in Clinical informatics, her students are also training a Cancer model and a Stem cell model, deploying Systems Biology approach and other Gene Networking BioInformatics tools. This novel area of research will hopefully lead to further understanding the tipping of balance from a stem cell/normal cell to a transformed cancer cell. Owing to her immense interest in science journalism and writing potential, she is now on the editorial board of five-six International Journals. Her Specialties Include: Research/Teaching/Mentoring/Science-Journalism.Kim, J.-K. and Seo, S.J.AbstractApoB peptide 29mer was LDLR binding element in natural low density lipoprotein. LDLR is abundant in BBB, and overexpressed in malignant brain glioma. ApoB-conjugated gold nanoparticles (ApoB@AuNP) were synthesized to develop nanobeacon in Coulomb nanoradiator (CNR) therapy for the treatment of malignant glioma. Transcytosis to BBB and cellular uptake were tested using TEER method and ICP-MS, respectively, and compared with bare gold nanoparticles. In order to present nanoradiator-mediated electron/X-ray photon emission, SH-PEG-hydrocynine 5.5 was conjugated to ApoB@AuNP. ROS-enhancement of hydrocyanine-conjugated ApoB@AuNP was measured in situ under traversing proton beam based on ROS-oxidant fluorescence of hydrocyanine.BBB-transcytosis of ApoB@AuNP increased dose-dependently, demonstrated 3.4 fold augmentation compared to bare AuNP in 24 h post-incubation. Intracellular uptake in F98 glioma cell increased greatly by 10\u201320 fold compared to bare AuNP. CNR-mediated enhancement of ROS production was increased by 3.5 fold under 4-Gy traversing proton irradiation compared to non-treatment. In conclusion, ApoB@AuNP exhibited great potential as therapeutic nanobeacon with transcytosis-based BBB crossing and targeted delivery to malignant glioma for CNR therapy.BiographyProfessor Jong-Ki Kim is the chairman of Biomedical Engineering in School of Medicine, Catholic University of Daegu, based in Daegu City, South Korea. Professor Kim has published distinguished papers on the subject of Coulomb nanoradiator therapy and related physics. He is a recognized pioneer in the use of nanoradiator effects, giving many invited papers at international meetings around the world.After obtaining Ph.D. in Biophysics from the State University of New York at Buffalo in 1992, he spentpostdoctoral at Memorial Sloan-Kettering Cancer Center on NMR spectroscopy on DNA duplex containing photoadducts to reveal structural basis of DNA mutagenesis. He continued his research in MRI and MR spectroscopy at Catholic University of Daegu as Professor in School of Medicine since 1995. During last decade his research subject was extended to nanomedicine in the field of radiation impact on high-Z nanoparticles. Recently, he had spent visiting professor in Bioengineering, UC Berkeley. He pioneered in the development of site-specific Coulomb nanoradiator therapy using traversing Bragg-peak ion beam and targeted high-Z nanoparticles. Dr. Seo obtained Ph.D. in 2012 under guidance Prof Kim. Dr. Seo developed diffraction based synchrotron X-ray brain imaging and Coulomb nanoradiator therapy in a number of disease models including thrombosis and malignant gliom. He is currently a Research Professor in Biomedical Engineering at School of Medicine, Catholic University of Daegu.In, S.-I.AbstractAcupuncture has long been accepted as an effective therapy for the treatment of many functional disorders, such as pain and psychiatric disorders including anxiety and drug abuse. The invention of acupuncture as a therapeutic treatment is traced as far back as 6000 B.C., originating with the insertion of sharpened stones at specific acupuncture points. The ancient use of sharp stones as an acupuncture device is replaced by that of fine nee-dles made from various materials including bamboo, ceramic, bone, and plant thorns, with these in turn replaced by metal acupuncture needles, including those of gold, silver, copper, and stainless steel. The biological basis of acupuncture still remains unclear, however a considerable number of studies has established a general concept that acupuncture contributes to the neurochemical balance in the central nervous system (CNS) and recovery or maintenance of homeostasis via interactions between needles and the surrounding tissues. In our previous study we reported the activation of the A-beta afferent fiber (sensory nerve fiber) of the ulnar nerve promoting cellular activation by acupuncture at Shenmen (HT7) points for modulating cocaine-induced addictive behavior. Moreover, it is found that mechanoreceptors in the superficial and deep afferents of the ulnar nerve play a functional role in producing acupuncture effects during mechanical stimulation of HT7. Involvement of the afferent fibers in acupuncture is supported by studies investigating acupuncture-like stimulation of superficial or deep tissues for reducing micturition contraction of the urinary bladder and acupuncture analgesia abolished by blockade of afferents fibers from muscle. In acupuncture therapies manual manipulation of acupuncture needles is still the most practicable clinical procedure to enhance the stimulation intensity for improved therapeutic effects. Various needle parameters such as diameter depth of insertion number of needles used per session and needle surface modification have been investigated for improved acupuncture performance. These studies suggest that employing.BiographyProfessor SU-IL IN is the Dean of External and International Affairs at DGIST (Daegu Gyeongbuk Institute of Science and Technology) since 2016. He has been working at DGIST since 2012. He received his Ph.D. in Chemistry from the University of Cambridge in 2008. He then became a postdoctoral research associate at the Technical University of Denmark in 2010. He also joined the Department of Chemistry at Pennsylvania State University as a postdoctoral fellow before joining DGIST. Professor In\u2019s current researches include synthesis and analysis of functional nano (bio)-materials for environmentally friendly renewable energy such as photovoltaic, heterogeneous catalysis and biocatalysts. A central goal of this work is relating surface structure/properties, size and composition to the catalytic activity and microbial fuel cell (MFC).Nie, L.AbstractBiofilms on cutaneous skin wounds and biomaterials implants are hard to eradicate with antibiotics due to antibiotic resistance and the lack of new antibacterial agents. Such biofilms could be broken by using the nanotechnology, without resorting to the antibiotics, such as metal-based nanoparticles. Poly (acrylic acid) (PAA) functionalized iron oxide nanoparticles were synthesized through a two-step process in this paper. Firstly, Oleic acid-coated nanoparticles were synthesized via a thermal decomposition of iron oleate, then, through ligands exchange, PAA-coated iron oxide nanoparticles were synthesized with using THF as solvent. Finally, the toxicity of PAA-coated iron oxide nanoparticles on skin cells and efficacy of nanoparticles against bacteria were evaluated. The results showed that the HaCat cell had a good viability while culturing with nanoparticles for 3 days, moreover, the PAA-coated iron nanoparticles caused bacteria killing. The PAA-coated iron oxide nanoparticles we prepared might be the ideal antibacterial treatment, had a better activity than iron oxide nanoparticles alone.BiographyDr. Lei Nie obtained the Ph.D. in nanotechnology and Science from Huazhong University of Science and Technology in 2013, after that, he started work as an assistant professor in Ningbo Institute of Materials Technology & Engineering, Chinese academy of science, during his time in Ningbo, he continued his research in tissue engineering, growth factors delivery system, and biodegradable copolymer functionalization. In 2015 he moved to Free University of Berlin in Berlin, Germany where he focused his research on nanoparticles preparation and functionalization for biofilms application, and distinguish the influences of hydrophilic surface functionalization and transfer in aqueous media on the magnetic properties and the oxidation state of iron oxide nanoparticles. He is currently an associate professor in Xinyang Normal University at China, he focuses his researches on polymer synthesis for multi-function hydrogels, nanoparticles functionalization for drug delivery, biodegradable scaffold for 3D cell culture and tissue engineering.Cramer, J.Abstract\u00ae) is an evaluation and treatment intervention designed to restore musculoskeletal asymmetries by correcting restricted ROM, speed and stability of movement, and sensations such as pain or tightness in a quicker hands-free manner.Traditional orthopedic evaluations often lead to inaccurate diagnoses of musculoskeletal injuries as clinicians tend to gear their evaluation and treatment plan towards the local symptomatic area. Applying this process may generate misdiagnoses or involve prolonged unnecessary treatments away from the actual problem. These concerns especially arise with the intricacies of the shoulder complex. Glenohumeral (GH) range of motion (ROM) asymmetries associated with total rotational motion (TROM) deficits in overhead athletes demonstrated increases in injury rates. TROM is the unilateral sum of GH internal/external rotation. TROM deficit occurs when there is an asymmetrical difference greater than 5\u00b0. Pathologic GIRD (pGIRD) occurs when IR in the dominant shoulder is greater than 18\u201320\u00b0 and a bilateral TROM asymmetry greater than 5\u00b0. Common treatment interventions consist of static stretching focused on increasing IR and typically range from four to six weeks. Regional interdependence (RI) is a concept explaining how seemingly unrelated impairments may be directly or indirectly related to the patient\u2019s primary complaint. Utilizing this concept, Total Motion Release , and both are enhanced in subjects with PD than in the controls (p < 0.05). Medication decreased the reflex component of rigidity (p < 0.01). It is concluded that both reflex and intrinsic factors are responsible for rigidity. Present findings are clinically significant as they may provide guidance in development of effective therapeutic interventions. Further, this line of research is currently in progress by examining effectiveness of physical therapy programs (such as muscle stretching and therapeutic taping) on reducing neural reflex and intrinsic components in parkinsonian rigidity.Parkinson\u2019s disease (PD) is a progressive neurodegenerative disease characterized by rigidity, bradykinesia, resting tremor, and postural instability. Rigidity, defined as an increased resistance to passive movement of a joint, progresses faster than other motor signs in PD. Rigidity is attributable to both exaggerated neural reflex and altered muscle mechanical properties. However, little is known about the contributions of individual components to rigidity. Further, there is no evidence regarding the effects of dopaminergic medication on individual components. Objectives of this study were to quantify the contributions of neural reflexes and intrinsic muscle properties to rigidity, and investigate the effects of medication on each contributing component. Joint torque and muscle activities of the wrist in 14 patients and 14 controls were measured during externally-induced movements. Each subject with PD was tested in Off- and On-medication states. A system identification technique was applied to differentiate and quantify the neural reflex and intrinsic mechanical components. A mixed model of analysis of variance (ANOVA) was performed to compare the differences between the two components of rigidity for both groups, and to compare between the Off- and On-medication states for patients. The results showed that reflex and intrinsic components are comparable in Neurophysiology from the University of Bristol, UK. Upon completing her doctoral study, Dr. Xia worked as a Research Scientist at the Institute of Neurology, University College London, UK. She also completed a post-doctoral research fellowship in neurological rehabilitation at Northwestern University\u2019s Feinberg School of Medicine and the Rehabilitation Institute of Chicago, USA. Dr. Xia has been a core faculty in Physical Therapy since 2003. Her current and previous teaching activities in physical therapy curriculum include motor control and motor learning, neuroscience, critical inquiry and evidence-based practice, and expertise practice in physical therapy. In addition to teaching, Dr. Xia has mentored doctoral physical therapy students, research associates, and post-doctoral fellows through their research projects. Dr. Xia\u2019s research interests focus on motor control, sensorimotor integration, neural rehabilitation, and Parkinson\u2019s disease. Her research has generated numerous peer-reviewed articles published in prestigious journals such as Clinical Neurophysiology, Experimental Brain Research, Journal of Physiology (London), and Physical Therapy journal. Her publications are widely cited by investigators worldwide. She has also served as the principal investigator of several funded projects\u2014including a few sponsored by the National Institutes of Health (NIH), USA. She is the recipient of several Research Awards and Fellowships.Lenger, K.AbstractThe nature of homeopathy using dilutions beyond the Avogadro number is clarified since Lenger\u2019s detection of magnetic photons in the MHz-region of homeopathic remedies by two different magnetic resonance methods: by the Tesla-flatcoil system and delayed luminescence (DL) using a modified photomultiplier. Separation of the newly detected magnetic photons from their carrier substance ethanol or saccharose globules took place when the measuring system has a bigger resonating frequency field than the field between carrier substance and photons. Characteristic frequency spectra and the potency levels were measurable by the number of photons, taking DL or by the characteristic size of the magnetic frequency field separating the photons in dependence on the potency level and on the potentized substance. Six unknown remedies could be identified by DL. The Law of Similars (Hahnemann 1796) can be expressed like this: the frequencies of the patient must match the frequencies of the remedies to get in resonance: either to attenuate the ill making frequencies or to enhance the amplitude of both. Since Einsteina human body is an electromagnetic wave package. It is assumed that healing takes place firstly quantumphysically on the energy level; later on the pathological pathways are regulated. This is in agreement with Popp\u2019s quantumphysical theory about health and illness and F\u00f6rster saying that each chemical reaction takes place on a higher energetic level of the atoms. The uptake of photons is necessary in biological systems to keep them alive in steady-state.Because of its efficacy according to the resonance principle homeopathy is a regulation therapy curing both: hyperfunction and hypofunction of a pathological pathway and consciousness applying additional potentized substrates, inhibitors and enzymes: Hashimoto disease supported by the substrates of the thyroid-pathway, asthma was healed by using the highly potentized substrates of the respiration chain and those of the glycolysis, paralyses require mostly the neurotransmitter Acetylcholine which biosynthesis is supported by highly potentized substrates and inhibitors of glycolysis and fatty acid oxidation. The psychological problems of the patients are simultaneously healed by the applied remedies. Homeopathic healing means that body and consciousness are treated as one because homeopathy is a holistic medicine.BiographyKarin Lenger studied Biochemistry at the universities of T\u00fcbingen and Cologne/Germany. She worked as a Scientific Assistant at the Medical University of L\u00fcbeck/Germany for 12 years performing her biochemical enzymatic studies: enzymatic gene regulation, cancer research, enzymatic mechanisms of steroid hormones. She started her homeopathic career as a Lecturer for classical homeopathy at DHU (Deutsche Hom\u00f6opathie Union = German Homeopathy Union) in Karlsruhe 1986\u20131994. Since 1995, Dr. Lenger has been working as a practising doctor and Lecturer for classical homeopathy in Europe. Over the years she has developed the \u201cbiochemical homeopathy\u201d by using substrates of pathological enzymes in high levels of potentization. She detected photons in high homeopathic potencies by scientific proof, by two magnetic resonance methods. She has developed a model of physical and biochemical function of homeopathy. She has a lot of publications and had been invited for speeches on many world congresses in Europe, USA, Shanghai, Dubai, Hongkong, Nangjing.Reed, K.S.AbstractThe number of people with gait dysfunction is fast growing due to population growth, war, ageing and accidents. The cost of orthopedic devices essential to restore function and improve quality of life is not affordable for many. Earlier research in this area suggests that bamboo is a suitable material for orthopedic appliances, especially exoskeleton. In order to further advance research in this area, the role of bamboo as a structural material, with special emphasis on orthopedic appliances was carried out in this work. It was found that bamboo is being extensively used as infrastructure material in Asia and Africa due to its excellent mechanical properties. Its usage has ranged from simple fence construction to bridges. The use of bamboo as an orthopedic rehabilitation aid is prevalent in developing world where traditional bonesetters have utilized bamboo as a method of splinting for fracture treatment and management. Despite the great progress made in the usage of bamboo as infrastructure materials, the application in orthopedic rehabilitation appliances is limited to traditional practices due to a lack of standardization. Our work revealed that bamboo can be used in a variety of ways for orthopedic purposes, and can be classified into three categories based on the complexity of design and manufacturing.BiographyKischa S. Reed, PT, DPT, COMT is an Assistant Professor in the Division of Physical Therapy since 2012. Dr. Reed received her B.S degree in Physical Therapy from the Florida Agricultural and Mechanical University in 1998 and earned her transitional Doctor of Physical Therapy degree from Utica College in 2011. As a Certified Orthopedic Manual Therapist (COMT), Dr. Reed is able to combine the latest Evidence-Based research in Manual Therapy/Physiotherapy with the Maitland Approach of assessment and treatment which emphasizes clinical decision-making, advanced orthopedic clinical practice, accurate techniques of assessment/treatment, effective treatment progression and safety, to provide advanced treatment techniques in acute, chronic, and post-surgical spine rehabilitation. Having this knowledge of manual skills and core competencies align well with her current teaching responsibilities in Analysis of Human Motion (Biomechanics/Kinesiology), Orthopedic Physical Therapy, Vertebral Manipulation and Mobilization, and Physical Agents in the physical therapy program. Dr. Reed is a member of the Florida Physical Therapy Association (FPTA), where she formerly served as a Florida Assembly representative.Gurses, S.Abstractperceived-self through a redundant motor system by fusing senses relaying different originated information in a variety of domains; such as time, frequency, and symbolic spaces. The complex dynamics have been investigated by constructing its phase-space representation from Antero-Posterior Center-of-Pressure (xCoP) trajectories, where a characteristic pattern has been identified depending on nonlinear dynamics (slightly positive largest Lyapunov exponent causing deterministic chaos), e.g., caused by sensory thresholds demonstrating individual signature properties. Furthermore, correlation dimension estimates computed from xCoP trajectories have been shown to reveal that human quiet standing demonstrates multiple degrees of freedom dynamics having a fractal structure with a considerable level of noise embedded in the signal . On the contrary, Bilateral Vestibular Loss patients\u2019 (BVL) xCoP trajectories presented comparatively poor dynamics having lost some characteristic low frequency-bands in their spectral analyses against some speedy frequency-bands gained , whose unaltered presence has been proposed (through thermodynamics based nonlinear metrics) to work as a potential for ecological source of adaptive information.Human postural sway demonstrates complex dynamical characteristics where postural control system utilizes the BiographyDr. Senih G\u00fcrses having graduated from Medical College of Hacettepe University, Ankara in 1986, worked as a physician for about 10 years while completing his MSc Thesis in Biomedical Engineering at Bosphorus University, \u0130stanbul and the University of Tokyo. He completed his Ph.D. Thesis on Postural Dynamics and Stability at Middle East Technical University, Ankara in 2002 and moved to Rehabilitation Institute of Chicago as a post-Doctoral Researcher. There he studied nonlinear dynamical characteristics of human postural control, especially sensory-motor performance investigated in healthy versus vestibular loss subjects and cerebellar patients. In 2005, he returned to Middle East Technical University as an Assistant Professor and lectured in Principles of Mechanics and Biomechanics. From there on, a postural research team established by Dr. G\u00fcrses collaborating with Oto-Rhino-Laryngology and Neurology Departments of G\u00fclhane Training and Research Hospital and Physical Medicine and Rehabilitation Department of Gazi University Medical College studied different aspect of sensory fusion and multi-integration, perception/action in postural control in healthy and diseased conditions especially in view of ecological adaptation revealed through thermodynamically based nonlinear dynamical metrics utilized by information (ergodic) theory.Chen, J.-H., National Yang-Ming University, TaiwanCho, K. Yonsei University, KoreaLitscher, G. President of ISLA , Medical University of Graz, AustriaCaesar, I. Matrix City LLC, RussiaMei, L. Beth Israel Deaconess Medical Center, USAWen, G. South China University of Technology, ChinaSegura-Saldana, P. Universidad Peruana Cayetano Heredia, Per\u00faXu, S. Peking University, ChinaLee, C.-J. Fu-Jen Catholic University, TaiwanHeer, R. Austrian Institute of Technology, AustriaSimone, G. Northwestern Polytechnical University, USASingh, A. Harvard University, USAStanciu, G.A. University Politehnica of Bucharest, RomaniaSong, W. Shenzhen University, ChinaSun, R. Chairman & CEO, Xi\u2019an Pillar Bioscience Co. Ltd., ChinaPinchuck, L. President & CEO, Innovia, LLCL\u00f3pez-Bayghen, E. Cinvestav-IPN, M\u00e9xicoCedillo, L. Cinvestav-IPN, M\u00e9xicoOcampo, A. Cinvestav-IPN, M\u00e9xicoCamargo, F. Cinvestav-IPN, M\u00e9xicoFrighetto, L. Hospital Moinhos de Vento, BrazilWeinshilboum, R.M. Mayo Clinic College of Medicine, USAWang, L. Mayo Clinic, USABou-Sa\u00efd, B. LaMCoS-INSA Lyon, FranceMilia, P. Italy University of Perugia, ItalyLee, Y.-H. National Central University, Taiwan Wang, S. Northwestern Polytechnical University, ChinaTeng, J. Louisiana State University Health Sciences Center in Shreveport, USAHerrera, G. Louisiana State University Health Sciences Center, USATurbat-Herrera, E. Louisiana State University Health Sciences Center, USAChen, H.H. Founder and CSO, StemEasy Biotech Ltd., ChinaPurwati Stem Cell Research and Development Center, Universitas Airlangga, IndonesiaYang, Y. The University of Hong Kong, Hong KongLi, J. Zhengzhou University, ChinaNordon, R.E. University of New South Wales, AustraliaChen, H. The Affiliated Drum Tower Hospital of Nanjing University Medical School, ChinaWang, H. East China Normal University, ChinaHuang, W.-X. Tsinghua University, ChinaShan, X. Shandong Normal University, ChinaWu, Z. Canterbury Pharmaceuticals, New ZealandZheng, W. University of Macau, ChinaXie, L. Molecular Medicine of University of Georgia, GeorgiaBhojwani, J. Indore University, IndiaKim, J.-K. Biomedical Engineering and Radiology, School of Medicine, Catholic University of Daegu, KoreaSeo, S.J. Biomedical Engineering and Radiology, School of Medicine, Catholic University of Daegu, KoreaIn, S.-I. DGIST, KoreaNie, L. Xinyang Normal University, ChinaCramer, J. Germantown Academy/Philadelphia Freedoms, USAXia, R. University of Saint Mary, USALenger, K. Institute for Scientific Homeopathy, GermanyReed, K.S. Florida Agricultural and Mechanical University, USAGurses, S. Middle East Technical University, Turkey"} +{"text": "Cell & Bioscience.Two research groups led by Dr. Jim Hu of University of Toronto, Canada and Dr. Renping Zhou of Rutgers University, USA, respectively, won the 2016 Ming K Jeang Award for Excellence in Cell & Bioscience in 2016, have been selected to receive the Ming K Jeang Award for Excellence in Cell & Bioscience. The Ming K Jeang Award for Excellence in Cell & Bioscience was established in 2011 with a generous donation from the Ming K. Jeang Foundation to honor outstanding research articles published in Cell & Bioscience, the official journal of the Society of Chinese Bioscientists in America . A committee of Cell & Bioscience Editors, chaired by Dr. Dong-Yan Jin, considered all research articles published in the journal in 2016 to select the following two articles to receive the award [We are very pleased to announce that two research groups, who each published an outstanding research article in Chan-Mi Lee, Sahil Gupta, Jiafeng Wang, Elizabeth M. Johnson, Leslie J. Crofford, John C. Marshall, Mohit Kapoor and Jim Hu.Cell & Bioscience 2016 6:43.Gitanjali Das, Qili Yu, Ryan Hui, Kenneth Reuhl, Nicholas W. Gale and Renping Zhou.Cell & Bioscience 2016 6:48.Congratulations to these two groups of investigators for jobs well done!We are looking forward to receiving contributions of outstanding research articles from the scientific community in 2017 and beyond."} +{"text": "The correct email address is: There are errors in the Funding section. The correct funding information is as follows: The project has been sponsored in part by the National Center for Development (Poland) grant no POIG.01.04.00-22-140/12 issued to BioVentures Institute Ltd., in which the University of Gdansk has been a scientific partner. The BioVentures Institute Ltd. provided support in the form of salaries for the following authors: Piotr M. Skowron, Joanna Zebrowska, Lukasz Janus, Kasjan Szemiako, Edyta Czajkowska, Natalia Maciejewska, Malgorzata Skowron and Agnieszka Zylicz-Stachula. These authors are (or have been) also University of Gdansk employees or students. The specific roles of these authors are articulated in the \u2018author contributions\u2019 section. During the grant realisation, BioVentures Institute Ltd. also provided materials, reagents and equipment. BioVentures Institute Ltd. has played a predominant role in this study during all of its stages: the concept development, experiments design and realisation, as well as the decision to publish and the manuscript preparation. The project has also been sponsored in part by: University of Gdansk, Polish Ministry of Science and Higher Education fund no DS 530\u20138645-D691-17; National Centre for Research and Development (Poland) grant no STRATEGMED1/235077/9/NCBR/2014 and Gdansk University, Chemistry Department fund no BMN 538-8370-B817-17."} +{"text": "The research has also been supported by the European Union, co-financed by the European Social Fund (EFOP-3.6.2-16-2017-00013). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Information is missing from the Funding section. The complete, correct funding information is as follows: The research was carried out as part of the EITKIC 12-1-2012-0001 project, which is supported by the Hungarian Government, managed by the National Development Agency, financed by the Research and Technology Innovation Fund and was performed in cooperation with the EIT Digital Budapest Associate Partner Group ("} +{"text": "Interventions that improve maternity care for immigrant women in the UK: protocol for a narrative synthesis systematic review. BMJ Open 2017;7:e016988. doi: 10.1136/bmjopen-2017-016988Higginbottom GMA, Evans C, Morgan M, The following Disclaimer statement should have been included in the article:Disclaimer The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health."} +{"text": "Background: While documentation of clinical aspects of General Practice/Family Medicine (GP/FM) is assured by the International Classification of Primary Care (ICPC), there is no taxonomy for the professional aspects (context and management) of GP/FM.Objectives: To present the development, dissemination, applications, and resulting face validity of the Q-Codes taxonomy specifically designed to describe contextual features of GP/FM, proposed as an extension to the ICPC.Development: The Q-Codes taxonomy was developed from Lamberts\u2019 seminal idea for indexing contextual content (1987) by a multi-disciplinary team of knowledge engineers, linguists and general practitioners, through a qualitative and iterative analysis of 1702 abstracts from six GP/FM conferences using Atlas.ti software. A total of 182 concepts, called Q-Codes, representing professional aspects of GP/FM were identified and organized in a taxonomy.Dissemination: The taxonomy is published as an online terminological resource, using semantic web techniques and web ontology language (OWL) (http://www.hetop.eu/Q). Each Q-Code is identified with a unique resource identifier (URI), and provided with preferred terms, and scope notes in ten languages and search filters for MEDLINE and web searches.Applications: This taxonomy has already been used to support queries in bibliographic databases , to facilitate indexing of grey literature in GP/FM as congress abstracts, master theses, websites and as an educational tool in vocational teaching,Conclusions: The rapidly growing list of practical applications provides face-validity for the usefulness of this freely available new terminological resource. KEY MESSAGESA taxonomy for professional aspects of GP/FM is proposed as an extension to the ICPC classification.182 Q-Codes are available as an online semantic web resource in 10 languages.Face validity of this new taxonomy is illustrated by the rapidly growing list of current applications.Several attempts have been made to define the distinct concepts of general practice and family medicine (GP/FM), as in statements of professional organizations and mediFor the documentation of clinical data, the International Classification of Primary Care (ICPC-2) is used , often iICPC-2 has been developed to identify reasons for encounters, such as symptoms, acts performed or requested and diagnosis in daily practice. However, it was not constructed to reflect the professional context of GP/FM, such as organizational, managerial, ethical, environmental, educational, and investigational factors.The absence of adequate normalized keywords for identifying specific professional contextual features of GP/FM is an impediment to proper indexing of grey literature in GP/FM.More than 50% of the scientific outputs of general practitioners at congresses are never published ,15. SincA taxonomy of professional contextual features of GP/FM has been developed to address this issue using present day technology for information storage and retrieval , such asThis new taxonomy was named Q-Codes, in honour of the late Henk Lamberts, a Dutch professor of family medicine (University of Amsterdam), who proposed to use the letter Q (not yet used in ICPC) for the development of a rudimentary coding system for his departmental research library.The aim of this background paper is: (i) to shortly describe the development of the Q-Codes taxonomy, designed to cover the professional contextual aspects of GP/FM; (ii) to present its dissemination tools available for accessing the taxonomy; and (iii) to describe a number of recent applications, as an exploration of the face-validity of this new terminological resource for primary care.A qualitative and iterative analysis of 1702 abstracts in English or French from six GP/FM congresses held between 2007 and 2014 has beenhttp://atlasti.com/). Each abstract is read and code attributed along the classification ICPC-2 for clinical aspects and with a Q-Code for emerging professional contextual aspects. By grouping these identified concepts and their Q-Codes into subcategories and categories, gradually the taxonomy takes shape. Cimino\u2019s standard set of desiderata for terminological resources was applied [Starting point for the analysis of the congress abstracts were the original Q-Codes created by H. Lamberts. To identify what is really at stake and respect the bottom-up approach, only themes addressed by GPs during the conferences have been classified. The data was analysed in a grounded theory approach with Atlas.ti software .http://www.hetop.eu) [The taxonomy with 182 Q-Codes in 10 languages was published as a semantic web resource on the multilingual, multi-terminology portal HETOP after free inscription (etop.eu) and in uhttp://3cgp.docpatient.net for a constantly updated overview). We will limit ourselves here to describe examples of recent applications for three domains: facilitating bibliographic searching; facilitating indexing of congress abstracts and dissertations; and e-learning applications.Numerous practical are currently in use or under development , a French publications database and MEDLINE , which makes it difficult to conduct a search in MEDLINE. By clicking on the Q-Code QD8 on PubMed; .http://docpatient.net/3CGP/QC/clinical_exercise.htmTrainees in GP/FM have used this search engine to explore concepts at stake during a consultation, and to have a prepared bibliography search for each important theme used in daily practice. An example of this approach is given in http://ww.mgtfe.be (in French) and at the Department of General Practice of the University of Coimbra, Portugal). The results of long working hours devoted to the realization of an exciting final master thesis will remain unread, if not properly indexed. Students are offered ICPC, the Q-Code taxonomy, an indexing tool from the HeTOP portal, and a user guide to assist them in the task to index their work [Q-Codes taxonomy and ICPC-2 are used to index the master theses in GP/FM in French-speaking Belgium [This approach is now reused by researchers from Houston University, Texas, US, for indexing of question-answer pairs in telehealth Brazil and by the Brazilian Society of Family and Community Medicine (SBMFC) to support the indexing of the congress abstracts of the next SBMFC congress .Indexing of congress abstracts allows congress participants to identify more quickly particular contributions of interest during the conference. Researchers not attending the conference can also more easily pinpoint interesting contributions and their authors.In addition, indexing all abstracts allows for a careful study of the scope and main points of the conference. As an example, we present shortly the results of such an analysis of the 212 abstracts of the congress of the French Society of teachers in GP/FM (CNGE), held in Lille in 2014. ICPC-2 was used to index clinical issues, and the Q-Codes taxonomy for professional contextual features. The distribution of themes of the QT domain (training and teaching) gives an insight in the abstracts presented . In the In France, a multi-terminology concepts extractor (ECMT) is created for the full automation of the indexing process for conference abstracts, dissertations, and abstracts from the French LiSSa base . A projehttp://3cgp.docpatient.net/action-research/q-codes-in-an-e-learning-program-in-vietnam). In this project, natural language processing techniques are used to guide the students to the right Q-Codes for their documentation needs.In Vietnam, the Q-Codes taxonomy has been used in an e-learning programme for GP/FM students (http://www.hetop.eu/Q (free access) and in URI format . This taxonomy is available on the web on I format , as a prThis taxonomy and its tools of dissemination are available at the point of care, to practicing physicians, researchers, trainers and trainees in GP/FM, as an illustration of the extent and complexity of this profession.A growing list of recent applications of this taxonomy illustrates its rapid uptake, and provides face-validity to its usefulness for scientific communications in the field of GP/FM, by facilitating indexing of dissertations, master theses and congress abstracts, and facilitating bibliographic searches in grey literature repositories of primary care and in general bibliographic systems such as MEDLINE.The Q-Codes taxonomy has been developed from a strong empirical basis in a bottom-up approach, and applying terminological expertise and sound qualitative methods. To provide free access to the taxonomy, a database was published on the web, created with a standardized terminological structure and advanced semantic web techniques. Great effort was invested in linking and mapping the newly identified Q-Codes to existing concepts in existing terminologies, both linguistic (Babelnet) and conceptual (PubMed).Further ontological research is needed to determine whether the two main rules of taxonomical thinking have been respected: completeness and exclusivity (one place for each concept) . A taxonhttp://www.ph3c.org/Q) who are also the custodians of ICPC-2.The value of a taxonomy is determined by the quality and speed of the maintenance and updating process. Although Q-Codes have not been officially endorsed by the World Organization of Family Doctors International Classification Committee (WICC), the maintenance will be handled by a working group of WICC ,17. ThisOne could argue that the development of a profession-specific taxonomy for professional aspects of GP/FM is unnecessary, as the aims for such a taxonomy could be met with existing resources, such medical subject headings (MeSH). However, the experience gained during the development of the taxonomy has shown that many profession-specific concepts are not properly represented in MeSH . FurtherThe feasibility to translate the Q-Codes labelling and scope notes in ten languages illustrates that the taxonomy is robust to cultural diversity. The rapid acceptance of this new terminological tool and its integration in some recent applications is also an indication of face-validity. However, more in-depth research is needed to underpin the validity of this taxonomy for its intended uses. Studying the impact of the use of the taxonomy in search and indexing efficiency of medical students, general practitioners, librarians, and automated systems alike will be the next steps.The use of automatic ontology learning techniques, relying on term extraction and semantic relation extraction, should be explored to identify concepts and relations from literature and incorporate them in the taxonomy, and facilitate the maintenance process."} +{"text": "Common domains associated with life satisfaction in both young-old and old-old groups were transportation and social participation. Community and health services were associated with life satisfaction for the young-old group only. On the other hand, civic participation and employment was significantly associated with the old-old group only. Social participation is important for the young-old and the old-old. Ageing older adults can be a resource to the society. Implications for promoting and implementing age-friendliness were discussed in the context of successful and productive ageing and the need for a more refined taxonomy of social activities. Age-friendliness, promoted by the World Health Organization (WHO), aims to enable and support individuals in different aspects of life for fostering life satisfaction and personal well-being as they age. We identified specific aspect(s) of age-friendliness associated with life satisfaction and examined similarities and differences in age-friendliness and life satisfaction in young-old and old-old adults. Six hundred and eighty-two ageing adults were asked to complete a survey questionnaire consisting of the Age-friendly City Scale, Satisfaction with Life Scale, and sociodemographic variables. Multiple linear regression analysis was used to examine the effects of various domains of age-friendliness on life satisfaction among the young-old adults (aged 65 to 74, As in other Asian countries, Hong Kong is encountering increasing population ageing. According to the Hong Kong Population Projections Report, the population of adults 65 and over will increase from 15% in 2014 to 31% in 2044 [The age-friendly city concept is based on the framework for active ageing defined by the World Health Organization (WHO), rooted in the belief that a supportive and inclusive environment will enable residents to optimize health, participation, and well-being as they age successfully in the place in which they are living without the need to move \u20137. The eThe purpose of this study was to identify specific aspect(s) of age-friendliness associated with life satisfaction and examine similarities and differences in age-friendliness and life satisfaction in young-old and old-old adults. The concept of age-friendly city has based its formulation on active ageing \u20137. VarioNeugarten and colleagues considered age norms in regulating the life course . These nWith reference to successful or active ageing, there is a close association between subjective well-being and life satisfaction . Life saAccording to a study by Jeste et al. , despiteWith the emergence of Neugarten's view of young-old and old-old, more researchers have focused on these categories. However, existing studies have focused more on the young-old, since they shared some common characteristics. For instance, they are entering their first retirement and are relatively healthy, better educated, and active in social and political participation. One study has shown higher levels of well-being associated with volunteering among young-old adults . With reWhile previous studies have examined differences in ageing experiences between age groups of aged adults, to our knowledge, no studies have examined the specific relationship between age-friendliness and life satisfaction with reference to the young-old and old-old. Neugarten's model can have important implications for the understanding of how age-friendly concepts can contribute to the life satisfaction of aged adults. Therefore, the aims of the present study include (1) to examine the relationship between specific sociodemographic variables and life satisfaction among young-old and old-old adults, (2) to assess the relationship between specific domains of age-friendliness and life satisfaction with controlled sociodemographic variables, and (3) to identify similarities and differences in specific domains of age-friendliness associated with life satisfaction between young-old and old-old adults.Study participants were recruited mainly from community centres and nongovernmental organizations (NGOs) in Kowloon East Region including Kowloon City and Kwun Tong, using a convenience sampling approach. Inclusion criteria for participants included being Cantonese speakers, having comprehensive understanding without wearing a hearing aid, and being mentally sound. The original study aimed at exploring the age-friendliness of Hong Kong as perceived by individuals aged 18 or older. For the purpose of this study, we only select adults aged 65 and above for data analysis.All participants were asked to complete a structured questionnaire, which consisted of three main parts: the Age-friendly City Scale, the Satisfaction with Life Scale, and basic demographic variables.Based on the WHO age-friendly city guidelines, it was adopted and translated into Chinese . Eight dIt was used to measure the cognitive component of subjective well-being . It consSociodemographic variables including age, gender, education level, health status, marital status, income, and expenditure were used in the analysis as factors to be controlled statistically. These variables have been consistent in predicting life satisfaction in previous research . Gender Participants were introduced the background of our study by research helpers, who were trained master students and research assistants in the field of Psychology as well as members from the Institute of Active Ageing. After signing the informed consent form, a set of questionnaires would be given. A few aged participants had difficulties in reading or writing, and research helpers assisted these participants to complete the questionnaire by reading out the questions and asking them to denote their response to each item. Participants were given a supermarket coupon after completing the questionnaire.Multiple linear regression analyses were performed using IBM SPSS Statistics 23. It tested the direction and strength of the association between predictor(s) and dependent variable(s). The predictors of SWLS were formed into two blocks and analysed hierarchically using the ENTER method. The first block of predictors includes sociodemographic variables and were evaluated as statistically controlled factors of SWLS. The second block of predictors included dimensions of AFCS and their predictive effects on SWLS were evaluated with the sociodemographic variables adjusted. Differences in sociodemographic variables between young-old and old-old participants are shown in p < .05). There was no difference in scores for domains of civic participation and employment (p = .071) and communication and information (p = .091) between young-old and old-old participants. Both groups rated the social participation domain the highest and the community and health services domain lowest among the eight AFSC domains.The mean AFSC domain scores of young-old and old-old participants are presented in p < .05) with sociodemographic variables of expenditure (r = .36 and r = .34), income (r = .17 and r = .11), and health status (r = .24 and r = .39). Gender was significantly correlated with SWLS score only in the young-old group (r = \u2212.12). The results of the regression analysis show that gender, marital status, educational level, income, and expenditure explained a significant amount of variance in SWLS score in both young-old = 12.17, p < .01) and old-old = 16.04, p < .01) groups. In both groups, expenditure and health had significant positive regression weights on SWLS score, indicating that those who perceived they had more than enough to spend and better health status would show better life satisfaction.The results of analyses for the first research question, the association between sociodemographic variables and life satisfaction, are presented in Tables R2 = .14, F = 11.1, p < .01) and old-old = 8.95, p < .01) groups. Similarities and differences in domains of age-friendliness associated with life satisfaction between young-old and old-old groups were then addressed within the models. The domain scores for transportation and social participation were significantly associated with SWLS scores in both young-old and old-old, respectively. In addition, the domain score for community and health services was significantly correlated with SWLS scores only in the young-old group , while the domain score for civic participation and employment was significantly associated with SWLS only in the old-old group . In other words, young-old adults tended to be more satisfied in an environment with good transportation, social participation, and community and health services, while old-old adults were more satisfied with an environment with good transportation, social participation, and civic participation and employment.The results of analyses addressing the second research question, the association between AFCS and life satisfaction, are presented in Tables In this study, old-old participants reported significantly higher scores in most of the age-friendly city domains as well as the Satisfaction with Life Scale, compared to young-old participants. These findings support the idea of \u201cparadox of well-being,\u201d in which older adults maintain high subjective well-being even when facing different challenges . Old-oldVariables such as economic status and perceived health situation were positively correlated with life satisfaction in both young-old and old-old groups. These results support the findings of previous studies , 39\u201341. The main purpose of this study was to investigate the association between age-friendliness and life satisfaction after controlling for sociodemographic variables. In both young-old and old-old groups, all age-friendly domains were correlated with life satisfaction, and most of the domains were correlated moderately. With respect to the regression model, both transportation and social participation were significant correlates of life satisfaction for both young-old and old-old groups, indicating that aged adults are more satisfied if these two domains are fulfilled. Mobility and sociSocial and civic participation also emerged as major themes from the data generated from the eight focus groups involving eight-five older adults in Kwun Tong and Kowloon City , 49. ParMaking communities more age-friendly involves both physical and social infrastructure to enable older adults to participate in life-long activities in meaningful ways. Through personal and social engagement in the community, social inclusion is promoted and social capital is enhanced , 51. ThiCommunity and health services were significantly associated with life satisfaction for the young-old group but not the old-old. These findings also echo previous findings in Hong Kong Chinese population. Cheng measuredThe present study has also indicated suggestions for improvement in future research. Since the concept of age-friendly city was adopted from the WHO guidelines, some questions may not be pertinent in local context. For instance, cycle paths are not common facilities in all areas of Hong Kong. Moreover, volunteering may involve both social and civic participation . LevasseAge-friendliness is significantly related to the life satisfaction of the ageing population. Social participation is important for the young-old and the old-old. Ageing older adults can be a resource to the society. The findings of this study have delineated some of the similarities and differences of needs between young-old and old-old adults. Future directions will need to work on a more specific taxonomy of possible social activities and social participation and how they can take place in the living environment of aged adults to enhance their well-being and their contribution to the younger generations."} +{"text": "A total of 52 schools participated in the experimental implementation phase of the project P.A.T.H.S. . After completion of the Tier 1 Program (Secondary 1 level), 344 teachers and social workers responded to the Subjective Outcome Evaluation Form (Form B), assessing their views of the program and their own performance. Qualitative data analyses based on the schools' evaluation reports showed that the program implementers had enhanced knowledge and skills, learned to establish instructor-student relationships and cooperate with colleagues, and fostered self-development. The workers also appreciated the program philosophy and values, program design and resources, process of implementation, interaction between instructors and students, and program effectiveness. The findings also revealed that the workers encountered difficulties in the program implementation and they also made suggestions on how the program design, program arrangement, manpower deployment, and support for the program implementation could be improved."} +{"text": "AbstractPrionispachampaka Maulik, 1919 are described and figured in detail. The chaetotaxy of the head, mouthparts, legs, and dorsal and ventral surfaces of the body are given. The larva of P.champaka mine in the leaves of Polliajaponica Thunb. (Commelinaceae) and pupate in the base of the mid-ribs. The adults were also observed feeding on the leaves of Polliasiamensis (Carib.) Faden ex D. Y. Hong. The prominent diagnostic characters of immature stages of other species of the three genera of Oncocephalini are discussed.The last-instar larva and pupa of Prionispa Chapuis, 1875 is a member of the tribe Oncocephalini, Chapuis 1875 (Chrysomelidae: Cassidinae) and consists of 29 described species occurring in the oriental tropics , P.dentata Pic, 1938, P.houjayi Lee et al., 2009 (Taiwan), P.opacipennis Chen & Yu, 1962, and P.sincia Gressitt, 1950 (Fujian). This genus can be distinguished from the other genera in Oncocephalini, such as Chaeridiona Baly, 1869 and Oncocephala Agassiz, 1846, by the following characters: head with a distinct longitudinal ridge but without protuberance between the antennal bases; antennae not striate, third antennomere longer than the anterior two antennomeres combined; the labial palpi with three palpomeres; and the pronotum without tubercles infesting Polliathyrsiflora Endl. ex Hasskarl (Commelinaceae) (P.houjayi infesting Disporumkawakamii Hayata (Liliaceae) (P.champaka mining in the leaves of Polliajaponica Thunb. (Commelinaceae) in Jiangxi Province, China, and we also made some biological observations.So far only five beraceae , P.dentberaceae and Commlinaceae , P.fulvinaceae) , P.houjliaceae) , and Priulaceae) . In the Prionispachampaka were collected on wild plants that were placed in plastic zip-lock bags. Then larvae and pupae were reared and observed in the laboratory. Field-collected and laboratory-emerged adults were preserved as pinned specimens . One adult was collected at Jiulianshan National Nature Reserve in July 2016 (without host plant note), and one adult was collected at Bawangling National Forest Park in August 2016, on Polliasiamensis with feeding channels of adults.All immatures were collected at Anjishan Provincial Forest Park from 2015 to 2017, on PageBreakmacro lenses; dissection of heads and mouthparts were done using a Motic SMZ-140 and Olympus SZX2-ILLT stereomicroscope; figures and examination were obtained using an Optika B-292 microscope and Cannon EOS 70D camera. Descriptions of immature stages follows Three mature larvae, three pupae, and three pupal exuviae were examined morphologically. Larvae and pupae were preserved in anhydrous ethanol. For microscopic study, heads of the larvae were separated from the rest of body and then the mouthparts were dissected. The photos of adults were made using a Cannon EOS 7D camera and All studied material and adults were deposited at the Leafminer Group, School of Life and Environmental Science, Gannan Normal University, China.Mature larva and four setae on each side at base of leg with three short pointed setae laterally with three setae (one seta distinctly longer than others) and two campaniform sensilla ventrally, and numerous short spines dorsally. Maxillary palp (mp) with two palpomeres: first palpomere with one long seta and one short seta at apex, and one campaniform sensillum ventrally; second palpomere with a group of peg-like sensilla at apex. Mala with twelve long pointed setae apically. HypopharynxPageBreak (hyp) covered with numerous spines. Labial palp (lp) with one palpomere, with a group of small peg-like sensilla at the apex. Prementum (pre) with three setae on each side. Postmentum (post) with three short setae placed on each side medially.Maxillae and labium connate. Each stipes .lly Fig. . MesonotIn ventral view Fig. : head anspiracles at the base of the biting channel. Finally, the female covers this portion of the biting channel with feces for pupation (C.thailandica Kimoto (on the mid-rib on the upper surface of the leaf) (on the mid-rib on the upper surface of the leaf) (Dactylispa (unpublished data). The pupa of P.champaka can close its pupal mine with the broadened and flat caudal end of the abdomen, as can C.thailandica , O.promaf base) , C.thaihe leaf) , Platyprsurface) , Notosache leaf) , and somilandica ."} +{"text": "This study was aimed to find out epidemiologic characteristic of tuberculosis (TB) cases, and Human Immunodeficiency Virus (HIV) positive cases among TB patients (TB/HIV co-infection) through demographic, temporal, and spatial study in Urumqi.Descriptive statistics and multivariate logistic regression were applied to identify the epidemiologic characteristics and risk factors of TB epidemic and TB/HIV co-infection epidemic. All addresses of each TB case, TB/HIV co-infection case, and administrative street were transformed into geographical coordinate. Subsequently, the geocoded address for 82 streets was transformed into a dot map used as the basis of spatial datasets. In addition, the paper also used quantile map and the spatial scan statistic in order to identify the spatial distribution and spatial clusters of TB epidemic and TB/HIV co-infection epidemic.There was a declining trend of the notification rates of TB epidemic from 2007 to 2009, as well as a rising trend from 2010 to 2013. However, the notification rates of TB/HIV co-infection epidemic showed a rising trend from 2007 to 2010, and a declining trend from 2011 to 2013. Moreover, a significant share of TB epidemic and TB/HIV co-infection epidemic happened between the age of 15 to 45 years old, indicating an increase in risk of TB and TB/HIV infection. It is worth noting that the risk of HIV infection for male TB patients was 2.947 times than that of female patients. Han ethnicity and Uygur ethnicity in urban region accounted for a large proportion of total TB and TB/HIV co-infection cases. Most of the TB cases of minorities in Urumqi showed a statistically significant increase in risk of HIV infection than Han ethnicity in Urumqi. In addition, the spatial distribution of TB epidemic and TB/HIV co-infection epidemic was highly skewed. Most of the local clusters were located in urban area and rural-urban continuum where showed an increase in risk of TB and TB/HIV infection.The epidemiologic and spatial-temporal analysis of TB epidemic and TB/HIV co-infection epidemic demonstrates a potential connection between TB and HIV in Urumqi. Demographic, temporal, geographic factors are the reasons of causing TB and TB/HIV co-infection epidemic. Tuberculosis (TB) is contagious and airborne, ranking as a leading cause of death worldwide alongside Human Immunodeficiency Virus (HIV) infection . Despite2, which accounted for a large proportion of total TB and HIV cases reported in Xinjiang. In 2011, TB prevalence in Urumqi reached up to 1,526.12/100,000 [As the report of the Chinese Center for Disease Control and Prevention (CDC) pointed, Xinjiang province has been confirmed as one of the high TB and HIV burdened provinces of China, according to data collected during the period from 2010 to 2013 [/100,000 . At the /100,000 . The infHistorically, special risk factors conduced to high prevalence of TB and HIV micro-epidemics in specific areas in Urumqi \u20138. For eIn all infectious diseases research, it is important to evaluate whether observed cases are representative for general cases. Such spatial clustering analysis is an important implement for diseases spatial randomness tests. Furthermore, the spatial clustering tests would provide location information of local clusters and the risk of each cluster, used as the evidence of diseases prevention and control . Thus, iTherefore, the main objectives of this study were to: (1) use descriptive statistics and multivariate logistic regression to estimate the risk factors of TB epidemic and TB/HIV co-infection epidemic; (2) use GIS to explore the geospatial characteristic and examine data aggregated to the level of the street; (3) employ SaTScan tests to examine spatial clusters of TB epidemic and TB/HIV co-infection epidemic, along with the risk of TB and TB/HIV co-infection for each clustered street in Urumqi region.The study protocol was approved by Urumqi Center for Disease Control and Prevention. This article does not contain any studies with human participants performed by any of the authors. All patient information was anonymized and de-identified prior to analysis. Therefore, no ethics approval was required by our Investigation Review Board.Surveillance information for TB and TB/HIV is collated through the tuberculosis surveillance center and the Tuberculosis Registration Systems. Besides, CDC supplies all the case of TB, HIV diagnoses, with all tests following the principle of voluntariness. In the data set, it has covered the number of cases concerning HIV positive among TB people reported from 2007\u20132013 for each street in Urumqi. Therefore, the population data from Statistical Yearbook are able to be publicly available.Descriptive statistics were used to explore the demographic characteristics and temporal trends of TB epidemic and TB/HIV co-infection epidemic, which includes age, occupation, and ethnic group, etc. Multivariate Logistic Regression was performed to seek correlation between TB/HIV co-infection and risk factors such as gender , age, ethnic group, and living region . All p-values were two-tailed, and values less than 0.05 were considered statistically significant. The number of TB cases or TB/HIV co-infection cases was combined to examine spatial-temporal variations for each year and each street.In order to explore spatial variations for TB and TB/HIV co-infection epidemics and build spatial datasets, the paper transformed the geographical location of TB and TB/HIV co-infection cases into geographical coordination at first. The geocoded address for each street was then transformed into a dot map, with each dot representing the center of gravity for each street. BaiDu API assists to supply the longitude and latitude for each street and each case. Finally, the number of TB and TB/HIV co-infection cases was counted at street-level, used for connecting with spatial analysis dataset. Besides, for describing the spatial distribution of TB epidemic and TB/HIV co-infection epidemic, this paper applied quantile map, which along with the spatial databases are established by Geoda 1.6.7.Regarding the dimensions of space, the identification of the local clusters of TB epidemic and TB/HIV co-infection epidemic across different streets geographically was completed by SaTScan\u2122 v9.4.2, a spatial scan statistic developed by Kulldorff . The spa be the total number of cases in street i and its closest street j. And let U be the population size of cases in street i and its closest street j. In mathematical notation:i,j). Besides, it turned out that TB patients aged 30\u201345 had higher risk of HIV infection than other age groups, which is in accord with the conclusion of Peierdun MIJITI , Li WG . This maThe visualization of the TB epidemic and TB/HIV co-infection epidemic in Urumqi over a 7-year period showed that the study districts are geographically clustering. Meanwhile, according to the spatial analysis results, it indicated that TB epidemic and TB/HIV co-infection epidemic shared almost same spatial clusters during the period from 2007 to 2013. Socio-economic status of neighborhoods is associated with geographic clustering of TB and HIV related outcomes \u201322. The All results indicated that there is a close correlation between TB epidemic and HIV epidemic. Several studies have proved that HIV epidemic is strongly associated with TB at the early stages, which has become one key factor undermining global TB control \u201324. In tHowever, there are several limitations deserved to be discussed. Firstly, the data was extracted from the tuberculosis surveillance center and the Tuberculosis Registration Systems which failed to cover HIV/TB surveillance data. Secondly, there are several clustering analysis methods, and the choice of the methods might affect the result of disease clustering analysis . AlthougIn general, this paper explored the epidemiologic characteristic and the corresponding risk of TB and TB/HIV co-infection epidemics, which can provide an evidence for further socio-demographic predicting research on the TB and TB/HIV co-infection in Urumqi. Considering that the similar demographic characteristics as well as clustered districts of TB epidemic and TB/HIV co-infection epidemic have been observed, the related government organizations should carry out a wide-spread HIV screening on TB patients in Urumqi, for the purpose of improving the discovery rate of HIV on TB patients, providing in time prevention services for people with TB/HIV co-infection, and effectively reducing the impact of HIV on population health."} +{"text": "Reason for Corrigendum:In the original article, we have neglected to include Stephan Theiss . ST developed analyses programs to analyze hippocampal neuronal network activity and interpreted the data. His contribution to this work was supported by the German Ministry of Education and Research (BMBF: FKZ 031B0010B) and the European Union (EURO-TRANS-BIO project In-HEALTH). The authors apologize for this oversight. This error does not change the scientific conclusions of the article in any way.MP, GC, and FA performed the histological evaluations of organotypic slices. GC and AB performed the electrophysiological analysis of organotypic slices. TL and PW prepared organotypic slices cultures. KM and HS performed the capase-3/7 evaluations of organotypic slices. PW, AH, and BI collected human CSF, clinical evaluation and CSF analysis for the identification of normal CSF samples. SI performed the MEA-recordings and analysis of hippocampal neuronal networks. LA did critical revision of the manuscript and interpretation of the data. EH and SI conceived the study and wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. SI received financial support by EISAI to conduct research. However, the study design, data collection, data interpretation and data presentation were not influenced by EISAI."} +{"text": "Pseudomonas spp. and Acinetobacter spp., and to assess its usefulness in the routine work of the National Reference Centre for Susceptibility Testing (NRCST) in Poland. The evaluation of the Carba NP/CarbAcineto tests was carried out on a group of 81 Enterobacteriaceae, Pseudomonas spp. and Acinetobacter spp. isolates producing KPC-, NDM-, VIM-, IMP- or OXA-48, -23, -24/40, -58-type carbapenemases, and on 26 carbapenemase-negative strains cultivated on a broad panel of microbiological media. Subsequently, the performance of the Carba NP/CarbAcineto tests was assessed on 1282 isolates of Enterobacteriaceae, Pseudomonas spp. and Acinetobacter spp. from Polish hospitals, submitted to the NRCST during a 9-month period in 2014. The Carba NP/CarbAcineto results were compared with other phenotypic tests and/or polymerase chain reaction (PCR). The impact of the media on the results of the Carba NP/CarbAcineto tests was observed, with the Columbia blood agar yielding the highest sensitivity and clarity of the results. Furthermore, the Carba NP/CarbAcineto tests were included in the NRCST routine procedure for carbapenemase identification. The sensitivity and specificity of the Carba NP test were 95.8% and 93.3%, respectively, for Enterobacteriaceae, and 97.5% and 99.0%, respectively, for Pseudomonas spp. The sensitivity of the CarbAcineto test for Acinetobacter spp. was 88.9%. This study confirmed the usefulness of the Carba NP/CarbAcineto tests for the rapid detection of various types of carbapenemases.The aim of this study was to evaluate the Carba NP test (and CarbAcineto) for the detection of carbapenemases in Enterobacteriaceae, Pseudomonas spp. and Acinetobacter spp. has been observed all over the world in recent years. The predominant carbapenemases include \u03b2-lactamases of the Ambler class A (KPCs), metallo-\u03b2-lactamases (MBLs) of the class B and carbapenem-hydrolysing oxacillinases of the class D , and the specificity and negative predictive value (NPV) were 93.3% . For Pseudomonas spp. the sensitivity and PPV were 97.5% , and the specificity and NPV were 99.0% . For Acinetobacter spp., the sensitivity and PPV of CarbAcineto were 88.9% .The results of the comparison of the Carba NP/CarbAcineto tests with the standard methodology used by the NRCST, obtained with 1282 bacterial isolates suspected of carbapenemase production, are presented in Table Pseudomonas spp., with high sensitivity and specificity [The Carba NP test was described for the first time in 2012 by Nordmann et al. as a rapid, easy and cheap method for carbapenemase detection in Enterobacteriaceae and cificity . In prevcificity , 33\u201335. cificity \u201318, and Pseudomonas spp. and Acinetobacter spp. strains with various carbapenemases were used, along with a broad panel of commercially available media. The results clearly illustrated the influence of the media on the Carba NP/CarbAcineto results. According to our experience, the Columbia blood agar is the best choice for the cultivation of bacteria for the Carba NP/CarbAcineto tests, providing the highest sensitivity and clarity of the results. The VIM or NDM producers harvested from most of the MH agars yielded a remarkable number of false-negative results, which were most probably related to lower zinc ions\u2019 concentrations that is crucial for the MBL activity [Acinetobacter spp. [In the first part of this work, well-characterised Enterobacteriaceae, activity . Owing tactivity , 34\u201337. ter spp. , 38, 39 The Carba NP and CarbAcineto tests have been used in the NRCST from the end of 2013 and March 2014, respectively, and now are included in its routine algorithm for carbapenemase detection as reliable, inexpensive and easy screening tests, reducing remarkably the time for the first feedback information for clinical laboratories. Based on own experience, and in the context of the alarming spread of carbapenemase-producing Gram-negative pathogens in Poland, the NRCST has been recommending Carba NP/CarbAcineto tests for use in diagnostic microbiology laboratories all over the country."} +{"text": "The fourteenth author\u2019s name is spelled incorrectly. The correct name is: Jyotisna Rani.P. Jayaprakash is not listed in the byline. Dr. Jayaprakash should be listed as the eleventh author and affiliated with ICAR- Indian Agricultural Research Institute, Regional Station Wellington- 643231, The Nilgiris, Tamil Nadu. The contributions of this author are as follows: Performed the experiments, contributed reagents/materials/analysis tools, and helped in raising the 19,460 wheat accessions at IARI Regional Station.Amit Kumar Singh is not listed in the byline. Dr. Singh should be listed as the thirty-second author and affiliated with ICAR-National Bureau of Plant Genetic Resources, Pusa Campus, New Delhi, India. The contributions of this author are as follows: Performed the experiments.Z. Khan is not listed in the byline. Dr. Khan should be listed as the thirty-third author and affiliated with ICAR-National Bureau of Plant Genetic Resources, Pusa Campus, New Delhi, India. The contributions of this author are as follows: Performed the experiments."} +{"text": "Such breadth is supported by major national and international research funding, a range of industrial partners in the North East of England and beyond, as well as a large number of doctoral and post-doctoral researchers. The CSBB trains the next generation of scientists through a 1-year MSc in Synthetic Biology.The Centre for Synthetic Biology and the Bioeconomy (CSBB) brings together a far-reaching multidisciplinary community across all Newcastle University's faculties \u2014 Medical Sciences, Science, Agriculture and Engineering, and Humanities, Arts and Social Sciences. The CSBB focuses on many different areas of Synthetic Biology, including bioprocessing, computational design and A fundamental component of the CSBB is the Interdisciplinary Computing and Complex Biosystems (ICOS) research group, which carries out groundbreaking research at the interface of computing science and complex biological systems. Jointly led by Natalio Krasnogor and Anil Wipat, the group has internationally recognised know-how in synthetic biology. Since the first participation in the iGEM competition (2008) to the organisation of events such as the latest IWBDA , members of the ICOS group are present in major activities within the field. As part of the community involvement, the group holds the chair of the Synthetic Biology Open Language (SBOL) \u2014 an open standard format for the exchange of designs of engineered biological systems. Additionally, ICOS leads the MSc in Synthetic Biology of the CSBB, which trains the new generation of synthetic biologists.http://www.ncl.ac.uk/csbb/) in 2015, upon the appointment of Natalio Krasnogor as its director (JE and AW co-direct the centre). Researchers of over 10 institutions across the University (http://portabolomics.ico2s.org), a \u00a37.5 million initiative jointly funded by the Engineering and Physical Sciences Research Council (EPSRC), Newcastle University and industry, is a good example of such collaboration within the CSBB. The Centre has been instrumental in securing Newcastle's inclusion in major synthetic biology initiatives across the U.K., such as the Flowers consortium and the SynbiCITE Innovation and Knowledge Centre. Synthetic biology research is rapidly growing at Newcastle as shown by the recent appointments of Tom Howard, Jon Marles-Wright and Dana Ofiteru in biochemistry and metabolic engineering, and Angel Go\u00f1i-Moreno in computational synthetic biology and mathematical modelling.ICOS activities are split between the School of Computing Science and the Centre for Bacterial Cell Biology (CBCB), a research centre of the Institute of Cell and Molecular Biosciences. The CBCB, directed by Jeff Errington, is the world's first major research centre with a focus on fundamental scientific questions about bacterial cells. In it, over 20 groups collectively address a wide range of bacterial-based problems from different angles, including synthetic biology approaches. In addition, there are other centres on campus, such as the Policy, Ethics and Life Sciences (PEALS) research centre, which are involved in different aspects synthetic biology through the efforts of many world-leading scientist. In such a scenario, the need to co-ordinate all efforts in synthetic biology emerged with clarity years ago. In 2012, Jeff Errington and Anil Wipat established the CSBB under the name Centre for Synthetic Biology and Bio-exploitation to bring together academics with expertise in synthetic biology that would traditionally be found in different faculties/schools. The CSBB was renamed Centre for Synthetic Biology and the Bioeconomy new approaches, technologies and tools, (ii) applications and (iii) ethical, legal and social implications. The expertise available at the CSBB underpins the research carried out in Newcastle in many established themes .AgriculThe main facilities are located at the Centre for Bacterial Cell Biology (CBCB). It has world-class installations for all synthetic biology areas, including state-of-the-art laboratories. Microscopy facilities are specialised in fluorescence imaging allowing a wide range of measurements, such as 3D imaging, 4D multi-position, FRAP analysis or flow cell imaging. As a large contributor to the CSBB, the School of Computing Science provides cluster availability for computer-intensive analysis of large data sets. The Centre also integrates robotic facilities, since modern high-throughput biology requires a high degree of automation. The CSBB continually strives to provide top-of-the-range facilities, which currently include an Ion Torrent PGM genome sequencer and robots for the large-scale analysis of variations in yeast colonies.Microfluidics and lab-on-a-chip technologies are becoming a powerful tool for synthetic biology since they permit single-cell manipulation to an unprecedented extent. We built our microfluidics foundry with flexibility and scalability in mind, to ensure we can keep up with this rapidly evolving field.http://www.ncl.ac.uk/csbb/research/software-data-repository-resources/ and http://ico2s.org/resources.html.The members of the Centre have produced a large number (over 20) of Software tools and resources for Synthetic Biology that have been released to the community. Details are available at Over the last 5-year period, interdisciplinary funding secured by members of the CSBB amount to over \u00a318 million. Key funding bodies contributing to such a figure include the Engineering and Physical Sciences Research Council (EPSRC), the Biotechnology and Biological Sciences Research Council (BBSRC), the Natural Environment Research Council (NERC), the European Union and industry collaborations among others.The Authors declare that there are no competing interests associated with the manuscript."} +{"text": "In Australia, Indigenous people experience poor access to health care and the highest rates of morbidity and mortality of any population group. Despite modest improvements in recent years, concerns remains that Indigenous people have been over-researched without corresponding health improvements. Embedding Indigenous leadership, participation, and priorities in health research is an essential strategy for meaningful change for Indigenous people. To centralize Indigenous perspectives in research processes, a transformative shift away from traditional approaches that have benefited researchers and non-Indigenous agendas is required. This shift must involve concomitant strengthening of the research capacity of Indigenous and non-Indigenous researchers and research translators\u2014all must teach and all must learn. However, there is limited evidence about how to strengthen systems and stakeholder capacity to participate in and lead continuous quality improvement (CQI) research in Indigenous primary health care, to the benefit of Indigenous people. This paper describes the collaborative development of, and principles underpinning, a research capacity strengthening (RCS) model in a national Indigenous primary health care CQI research network. The development process identified the need to address power imbalances, cultural contexts, relationships, systems requirements and existing knowledge, skills, and experience of all parties. Taking a strengths-based perspective, we harnessed existing knowledge, skills and experiences; hence our emphasis on capacity \u201cstrengthening\u201d. New insights are provided into the complex processes of RCS within the context of CQI in Indigenous primary health care. Globally, Indigenous peoples experience significant health disparities . AborigiSystems and stakeholder capacity to participate in and lead CQI research in Indigenous primary health care is required to generate benefits , 8. Yet Centre for Research Excellence in Integrated Quality Improvement for Indigenous primary healthcare services (the Centre) has collaboratively developed a CQI-RCS model. This model draws on capacity strengthening , development of a values and ethics protocol and co-chaired by Veronica Matthews, an Aboriginal researcher, guided the development of the CQI-RCS Program . The Lead Group\u2019s composition reflected systems-based participatory action research and triavice-versa), and senior researchers mentoring less experienced researchers. The protocol provided direction for the Lead Group\u2019s approach to shared decision-making and activities, including creating space for dialog regarding matters of power and control over the CQI-RCS program.The Lead Group developed a values and ethics protocol to express how the CQI-RCS Program would address the six values of ethical research with Indigenous people and communities Table 11. The prCQI-RCS means to enhance capacities to conduct and use CQI research that is valued by and of benefit to Indigenous peoples, in the Indigenous primary healthcare setting, with the specific purpose of supporting integrated quality improvement and building on the collaborative platform of the Centre.CQI-RCS acknowledges existing strengths, knowledge and work. Through \u201call teach, all learn\u201d, CQI-RCS involves a mutual exchange between Indigenous and non-Indigenous researchers, research users, and communities to ensure sustained benefit from CQI research. CQI-RCS enables individuals, communities, organizations, services and broader systems to make informed decisions about, participate in, utilize, lead and generate CQI research.A program logic describes how a program\u2019s activities, impacts, outputs, and outcomes interact, to show the intended causal links . DevelopSecond, the Lead Group sought feedback from all members of the Centre\u2019s network regarding: current CQI-RCS successes and challenges; suggestions to support Indigenous leadership and participation in CQI research; and short-term and long-term outcomes that could inform the development of the program logic and the CQI-RCS Program.Third, the Lead Group, Centre research-leads and invited Indigenous leaders in RCS came together for a 1-day workshop, designed and facilitated by Karen McPhail-Bell and Veronica Matthews with the Lead Group, to develop a CQI-RCS program logic model. Fourth, the Lead Group presented the model to the Centre\u2019s bi-annual network meeting for feedback, prior to finalization.The program logic identifies CQI-RCS program priorities for implementation tied to short- and long-term outcomes, activities, participation, input, assumptions, and monitoring and evaluation indicators. The priorities are: indigenous co-leadership in the Centre\u2019s CQI research; improve and expand the Centre\u2019s RCS activities; strengthen CQI research networks and partnerships; and develop sustainable CQI research for improving Indigenous health through primary health care. A detailed program logic was produced for each of these priorities, which tied together in one overarching program logic map Figure . The proFour principles informed the CQI-RCS approach and provide a possible starting point for those seeking to implement RCS models in Indigenous primary healthcare contexts. The principles are discussed in detail below: (1) mutual/two-way learning; (2) Indigenous co-leadership as core; (3) sharing power and facilitating relationships; and (4) resourcing and continuous improvement.At the core of the \u201call teach, all learn\u201d motto is the valuing of Indigenous cultures, knowledge, and expertise alongside Western research and knowledge, and recognition that different kinds of capacities are to be developed in different people, processes, organizations, and systems. For example, in the Indigenous primary health-care context, individual capacities include knowledge, skills, and experience to: conduct and critically assess research; participate in all stages of the research; contribute to research-informed action; maintain respectful relationships; and facilitate culturally safe processes. Consistent with other mutual RCS approaches, we found that not all the same capacities need to be developed in all stakeholders, and that such capacities are influenced by worldview, knowledge, experience, and relationships , 34. TheIndigenous-led research refers to research that is led and driven by Indigenous researchers, practitioners, policy makers, and communities in partnership with community organizations or through collaborative approaches involving Indigenous community/ies at each stage of the research process . As a stA commitment to Indigenous co-leadership brings considerations and tensions to negotiate collectively, to produce mutually beneficial outcomes. Methodologies that seek to support and value Indigenous knowledge within CQI research, and involve Indigenous people and their interests, are necessary . For nonThe Lead Group provided a structure to enable Indigenous co-leadership of CQI-RCS and share decision-making and direction between diverse individuals and perspectives. The development process collectively identified and formalized the CQI-RCS priorities and approach, which provided momentum for power-sharing and involvement of more Indigenous people within the Centre. Respectful relationships underpinned these processes and structures, as reflected in the program logic Figure . StrengtResources are required to enable the communication, relationships, engagement and training that facilitate CQI-RCS. Likewise, resourcing and continuous learning are essential to enable co-leadership, which is generally an additional responsibility for busy Indigenous translators/leaders/researchers. While the Lead Group developed the CQI-RCS program model, implementation will be the responsibility of all within the Centre. Using a continuous improvement approach, the Centre can now test application of the CQI-RCS program logic across its research programs and activities and monitor progress against the CQI-RCS priorities.CQI-RCS must occur across multiple domains and levels to enable broader, systems-level engagement in health research at all stages, from research question formation to dissemination of outcomes. Our multi-level, systems view of CQI-RCS is consistent with other RCS models, such as that of Kahwa et al. at the expense of Indigenous-led research , 69. HowThis paper presents the development of a CQI-RCS model in Indigenous primary health care. The program logic provides a framework for CQI-RCS implementation, monitoring and evaluation of collaboratively determined priorities and outcomes. This is important because there are no existing models of which we are aware that articulate and inform CQI-RCS in Indigenous primary health care in this way. The model also points to the importance of mutual learning, Indigenous co-leadership, power-sharing and sufficient resourcing. The new knowledge generated through developing this model echoes calls of Indigenous Australians seeking research reform that benefits Indigenous peoples , 33, 57.Veronica Matthews, Research Fellow, Wingara Mura Leadership Program Fellow, The University of Sydney, University Centre for Rural Health; Co-Lead: Karen McPhail-Bell, Research Capacity Building Fellow, University of Sydney, University Centre for Rural Health; Kerry Copley, CQI Program Coordinator\u2014Top End, Aboriginal Medical Services Alliance NT (AMSANT); Louise Patel, CQI Program Coordinator\u2014Central Australia and the Barkly, AMSANT; Roxanne Bainbridge, Principal Research Fellow\u2014Indigenous Health, School of Health, Medical and Applied Sciences, CQUniversity; Michelle Redman-MacLaren, Adjunct Academic, Centre for Indigenous Health Equity Research, CQUniversity; Senior Research Fellow, College of Medicine and Dentistry, James Cook University; Deborah Askew, Research Director, Southern Queensland Centre of Excellence in Aboriginal and Torres Strait Islander Primary Health Care ; Shanthi Ramanthan, Post Doctorate Fellow\u2014Health Research Economics, Hunter Medical Research Institute; Nalita Turner, Researcher, Menzies School of Health Research; Ross Bailie, Director, University of Sydney, University Centre for Rural Health\u2014North Coast; CIA the Centre; Jodie Bailie, Research Fellow , University of Sydney, University Centre for Rural Health; Isaac Hill (January\u2013April 2017), Statewide Data and IT Coordinator\u2014CQI Unit, Aboriginal Health Council of SA; Janya McCalman (January\u2013April 2017), Senior Research Fellow, School of Health, Medical and Applied Sciences, CQUniversity.Co-Lead: KM-B conceived, drafted, reviewed, and finalized the manuscript for submission. All authors critically revised and approved this manuscript and, with the Lead Group (named in Acknowledgements), provided co-leadership for the Centre CQI-RCS work that forms this manuscript\u2019s content.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "We assessed the subsequent risk of cholelithiasis development in patients with inflammatory bowel diseases (IBDs) such as Crohn\u2019s disease (CD) or ulcerative colitis (UC). We identified 8186 patients who aged \u226520 years and were diagnosed with IBD between 2000 and 2010 as the study cohort. A total of 8186 patients without IBD were selected by frequency-matching according to age, sex, comorbidities, and the index date of diagnosis, and they were identified as the control cohort. To measure the incidence of cholelithiasis, all patients were followed up until the end of 2011. The risk of developing cholelithiasis, either gallbladder stone disease = 1.76, 95% CI = 1.34\u20132.61) or common bile duct (CBD) stones and intrahepatic stones , was higher for the CD cohort than for the non-IBD cohort after adjusting for age, sex, and comorbidities of hyperlipidemia, diabetes, liver cirrhosis, hypertension, chronic obstructive pulmonary disease, stroke, coronary artery disease, and hepatitis C virus infection. However, UC was related to the development of GSD but not to CBD stones and IHSs . Our population-based cohort study demonstrated that CD is related to the development of cholelithiasis, including GSD alone and non-GSD-associated cholelithiasis. However, UC is only related to the development of GSD alone. Cholelithiasis consists of gallbladder stone disease (GSD), intrahepatic stones (IHSs), and common bile duct (CBD) stones, which are distinguished based on the stone distribution in the biliary trees. Cholelithiasis is one of the most commonly encountered medical conditions in gastrointestinal departments, and it may lead to cholecystitis, cholangitis, pancreatitis, and biliary tract cancer. GSD is the major type of cholelithiasis, and the prevalence of cholelithiasis in Taiwan has been reported to be approximately 5\u201310% ,3. CholeInflammatory bowel disease (IBD) includes Crohn\u2019s disease (CD) and ulcerative colitis (UC). With enhanced disease awareness, improved diagnostic tools, and an increasingly westernized lifestyle, the incidence of IBD has rapidly increased in Asia . ExtrainCD has been shown to be reportedly related to the development of GSD in epidemiological studies. However, associations between CD and IHSs or CBD stones remain unknown ,13. MoreThe National Health Insurance (NHI) program of Taiwan was established in 1995 and provides comprehensive and universal health care coverage to approximately 99% of the 23.74 million residents of Taiwan . This stThe two cohorts in this retrospective cohort study were IBD and non-IBD cohorts. For the IBD cohort, we identified patients aged \u226520 years who were newly diagnosed with IBD (ICD-9-CM codes 555\u2013556), including UC (ICD-9-CM code 556) and CD (ICD-9-CM code 555), from 1 January 2000 to 31 December 2010, and who had complete age and sex information. The index date for the IBD cohort was set as the first IBD diagnosis date. Patients with a history of cholelithiasis (ICD-9-CM code 574) before the index date were excluded. The non-IBD cohort included individuals without IBD and without a history of cholelithiasis before recruitment. For each IBD patient, one non-IBD individual was selected and frequency-matched according to age (five-year range), sex, and baseline comorbidities of hyperlipidemia (ICD-9-CM code 272), diabetes (ICD-9-CM code 250), liver cirrhosis , alcohol-related illness , hypertension (ICD-9-CM codes 401\u2013405), chronic obstructive pulmonary disease , obesity (ICD-9-CM code 278), stroke (ICD-9-CM codes 430\u2013438), coronary artery disease , hepatitis B virus infection , and hepatitis C virus (HCV) infection . The index date assigned to the non-IBD individual was the same date as that of the corresponding patient with IBD. All included patients were followed up until cholelithiasis diagnosis, withdrawal from the NHI program, death, or 31 December 2011. Emigration from Taiwan and death were the major reasons for withdrawal from the NHI program, and the analysis included both cause-specific and non-cause-specific deaths when the causes were identified and registered in the National Health Insurance Research Database (NHIRD) of Taiwan.t-test for continuous variables. To assess the difference in the cumulative incidence rates of cholelithiasis between the IBD and non-IBD cohorts, Kaplan-Meier analysis and the log-rank test were applied. The incidence density rates of cholelithiasis were estimated by dividing the number of cholelithiasis cases by the number of person-years for each risk factor, and the rates were then stratified by age group, sex, and comorbidity. Univariable and multivariable Cox proportional hazard regression models were employed to examine the effect of IBD on the risk of developing cholelithiasis. The results are expressed as hazard ratios (HRs) with 95% confidence intervals (CIs). The multivariable-adjusted models included all of the statistically significant covariates that were identified in the univariable model. Further analysis was performed to determine whether patients with IBD exhibit significant risks of developing cholelithiasis, including GSD and non-GSD, or various types of IBD, including UC and CD. All statistical analyses were performed using the SAS 9.3 software package for Windows . The level of significance level was set at p < 0.05, and the tests were two-tailed.The distribution of age, sex, and comorbidities was compared between the IBD and non-IBD cohorts by using the chi-squared test for categorical variables and the Student\u2019s We identified 8186 patients with IBD and matched them with 8186 individuals without IBD according to age, sex, and comorbidities . The majp < 0.001). As the follow-up duration increased, the risk of developing cholelithiasis in the IBD cohort increased compared with that in the non-IBD cohort.The mean follow-up times in the IBD and non-IBD cohorts were 7.15 years (standard deviation (SD) = 3.22) and 7.14 years . Consistent with studies conducted in Western countries, most of the patients with IBD in our study were aged <49 years and female . HoweverThe association between IBD and cholelithiasis demonstrated in this study was unlikely observed by chance, because the multivariable Cox proportional hazards regression analysis employed to assess the association between IBD and cholelithiasis was adjusted for age, sex, and comorbidities of hyperlipidemia, diabetes, liver cirrhosis, hypertension, COPD, stroke, CAD, and HCV infection. Furthermore, the results consistently indicated that the contribution of IBD to the development of cholelithiasis was greater among middle-aged patients, men, and patients without comorbidities , even thNotably, age, rather than IBD, was the key risk factor for cholelithiasis, and the contribution of IBD to the development of cholelithiasis was nonsignificant for older adults in our study . PreviouAlthough more laboratory studies are required to ascertain the mechanisms underlying the association between IBD and the development of cholelithiasis, the pathophysiology of the contribution of IBD to the development of cholelithiasis may be as follows. First, bile acid malabsorption and depletion in the ileum due to impaired bile enterohepatic circulation caused by inflammation or ileostomy may predispose individuals to the development of cholesterol GSD . Second,Our study had several merits. This is the largest population-based study to demonstrate the association between IBD and the risk of developing cholelithiasis in the Asia-Pacific region. In addition, this study is the first to indicate that CD is closely related to the development of GSD alone and non-GSD cholelithiasis. Moreover, our study is the first to demonstrate that UC is related to the development of GSD alone but not to non-GSD cholelithiasis ,10. Our 2; the significance of obesity was minimal in our study because of its low prevalence in the IBD cohort. However, we acknowledge that the factors of age, sex, and comorbidities of hyperlipidemia, diabetes, liver cirrhosis, hypertension, COPD, stroke, CAD, and hepatitis C virus infection might be over-adjusted since hyperlipidemia, diabetes, liver cirrhosis, hypertension, COPD, stroke, and CAD could be other effects of the initial liver disease. Furthermore, IBD could be the consequence of initial liver disease and would affect the aHRs of IBD in contributing to the development of cholelithiasis. Second, although the relationships of the severity, extension, and management of IBD with the development of cholelithiasis have been controversial in previous studies, we could not assess these associations because the data were unavailable in the LHID2000 [Our study also had some limitations. First, some confounding factors may have been overlooked in this study due to the inherent unavailability of data regarding lifestyle, socioeconomic and educational status, genetic history, and definite body weight and height in the LHID2000, all of which can affect the development of IBD and cholelithiasis. For adjustment, we replaced body mass index with obesity and smoking habits with COPD diagnosis. Obesity was coded when the clinically recorded body mass index was >30 kg/mLHID2000 ,25. ThirOur population-based cohort study demonstrated a close relationship between CD and cholelithiases, including GSD alone and non-GSD-associated cholelithiasis. However, UC is only related to the development of GSD alone. Because IBD is a chronic disease, clinicians should survey the presence of cholelithiasis in patients with IBD when monitoring extraintestinal manifestations. Furthermore, more studies are required to ascertain the actual mechanisms of IBD that result in a predisposition to the development of cholelithiasis."} +{"text": "GBD 2016 DALYs and HALE Collaborators. Global, regional, and national disability-adjusted life-years (DALYs) for 333 diseases and injuries and healthy life expectancy (HALE) for 195 countries and territories, 1990\u20132016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet 390: 1260\u201313442017; . The full-text version of this Article has been updated so that the list of authors is displayed in the correct order, in line with the pdf version, rather than in alphabetical order. This correction has been made to the online version as of Oct 12, 2017."} +{"text": "India State-Level Disease Burden Initiative Collaborators. Nations within a nation: variations in epidemiological transition across the states of India, 1990-2016 in the Global Burden of Disease Study. Lancet 2017;390:2437\u201360\u2014In this Article , changes have been made to the list of India State-Level Disease Burden Initiative Collaborators and affiliations. This correction has been made to the online version as of Nov 30, 2017, and the printed Article is correct."} +{"text": "The decade following the Regional Strategic Framework for Visceral Leishmaniasis (VL) elimination in 2005 has shown compelling progress in the reduction of VL burden in the Indian subcontinent. The Special Programme for Research and Training in Tropical Diseases (TDR), hosted by the World Health Organization (WHO) and other stakeholders, has coordinated and financed research for the development of new innovative tools and strategies to support the regional VL elimination initiative. This paper describes the process of the TDR\u2019s engagement and contribution to this initiative.Multiple databases were searched to identify 152 scientific papers and reports with WHO funding or authorship affiliation around the following 3 framework strategies: detection of new cases, morbidity reduction, and prevention of infection. TDR has played a critical role in the evaluation and subsequent use of the 39-aminoacid\u2013recombinant kinesin antigen (rK39) rapid diagnostic test (RDT) as a confirmatory test for VL in the national program. TDR has supported the clinical research and development of miltefosine and single-dose liposomal amphotericin B as a first-line treatment against VL. TDR has engaged with in-country researchers, national programme managers, and partners to generate evidence-based interventions for early detection and treatment of VL patients. TDR evaluated the quality, community acceptance, and cost effectiveness of indoor residual spraying, insecticide-treated bed nets, insecticide-impregnated durable wall linings, insecticidal paint, and environmental management as tools for integrated vector management in reducing sandfly density.TDR\u2019s engagement with country policy makers, scientists, and clinicians in the development of effective diagnosis, treatment, case detection, and vector control represents an important example of TDR\u2019s stewardship toward the elimination of VL in the Indian subcontinent. Since the early days of the kala-azar elimination programme in the Indian subcontinent, TDR has engaged with national control and research institutions to conduct research aimed at informing country policy and practice to identify and treat cases and to prevent transmission of the infection. This includes the evaluation of the rK39 rapid diagnostic test for the diagnosis of VL, the clinical development and evaluation of first-line treatments , the generation of evidence-based tools for early detection and complete clinical management of VL, the evaluation of the cost effectiveness of indoor residual spraying, insecticide-treated bed nets, insecticide-impregnated durable wall linings, and environmental vector management as tools for integrated vector management. The interaction and interdependence between implementation research, technical advice, partnership, and policy is yet another example of TDR\u2019s stewardship contribution and empowerment toward VL elimination in the Indian subcontinent. Continuing investment in translational research from the bench to the bedside to public health, established jointly by national control programmes, academics, and TDR coordinators, is imperative to block transmission and prevent a resurgence of VL in the future. Phlebotomous argentipes as the only vector species, and the availability of effective tools for diagnosis and treatment, all supported by historical evidence for the disappearance of VL in the 1970s following insecticide spraying for malaria eradication, favour the elimination of VL as a public health problem in the Indian subcontinent . Th. Th85]. A second multicentre research project supported by TDR was conducted to inform the integrated vector-management strategy of the Regional Strategic Framework for VL Elimination. A situation analysis in Bangladesh indicated that there was low community awareness that VL was transmitted through the bite of sandflies and could be prevented by vector control . Indoor The WHA 50.13 resolution and the Stockholm Convention calls for a reduced reliance on chemical pesticides, specifically DDT for vector control. Viable alternate strategies are needed for controlling vector-born diseases. As part of the integrated vector management, TDR research tested 3 interventions\u2014IRS, insecticide-treated bed nets, and environment management (EVM). IRS and, to a lesser and more variable extent, EVM , and long-lasting insecticide nets (LLINs) significantly reduced sandfly density , 116. CoBased on its ongoing research, TDR developed and field tested a monitoring and evaluation tool kit for IRS with indicators to monitor inputs , process , output , outcome (sandfly density), and impact (VL disease burden) 118]. T. T118]. The VL regional elimination initiative has been a daring, cooperative endeavour that has been possible because key tools and interventions were available and could be implemented. The elimination target is now either reached or within reach thanks to these instruments and the contributions of many actors at the national and international level. TDR-supported research has played a critical role in contributing to the development and selection of the essential diagnostics and treatments , to the development of strategies and approaches to identify cases and prevent transmission in different epidemiological settings , which have been adopted and rolled out by national programs, and to the development of tools to monitor the quality and impact of the VL elimination program. As countries rapidly progress toward VL elimination, TDR now focuses on transmission dynamics and integrated approaches that are feasible and sustainable in the long term to prevent the resurgence of VL.The overall scope of TDR is to support research to develop and validate cost-effective interventions and strategies for VL elimination in the Indian subcontinent while promoting country empowerment and research capacity through the training of dozens of in-country researchers and through learning by doing . TDR\u2019s uWHO created a regional policy environment and political commitment conducive for the elimination of VL from the Indian subcontinent. TDR\u2019s approach has been to create a partnership with research institutes and the control programmes of the 3 countries of the Indian subcontinent. Established in 2005, this collaboration with country-based researchers, national control programme managers, and other partners identified 3 broad thematic areas\u2014detect new cases at an early stage, reduce morbidity, and prevent infection\u2014for targeted intervention research aligned with the Regional Strategic Framework . A situaLeishmania, markers for progression to VL disease, the role of domestic animals in transmission, and the potential of PKDL as a reservoir for infection need to be better understood [As countries in the Indian subcontinent progress towards the elimination goal in the affected regions, the concern is that elimination may be mistaken for eradication, and both donor fatigue and programme complacency may drift attention to the next unfinished agenda . Limitatderstood , 125\u2013127derstood . The devderstood , better derstood , 128. DWderstood , 130. CoTDR engaged with national policy makers, scientists, and clinicians in the development and validation of strategies for elimination of VL in the Indian subcontinent.Linkage with the national programmes from the conception stage to identify and prioritize research needs facilitated the uptake of evidence-based interventions into national programme and policy.TDR\u2019s stewardship role in supporting intervention research, technical advice, training, and policy contributed to the elimination of VL in the Indian subcontinent.Cunningham J, Hasker E, Das P, El Safi S, Goto H, Mondal D, et al. A global comparative evaluation of commercial immunochromatographic rapid diagnostic tests for visceral leishmaniasis. Clinical Infect Dis: an official publication of the Infectious Diseases Society of America. 2012;55(10):1312\u20131319.Bhattacharya SK, Sinha PK, Sundar S, Thakur CP, Jha TK, Pandey K, et al. Phase 4 trial of miltefosine for the treatment of Indian visceral leishmaniasis. J Infect Dis. 2007;196(4):591\u2013598.Mondal D, Alvar J, Hasnain MG, Hossain MS, Ghosh D, Huda MM, et al. Efficacy and safety of single-dose liposomal amphotericin B for visceral leishmaniasis in a rural public hospital in Bangladesh: a feasibility study. Lancet Glob Health. 2014;2(1):e51-e57.Singh SP, Hirve S, Huda MM, Banjara MR, Kumar N, Mondal D, et al. Options for active case detection of visceral leishmaniasis in endemic districts of India, Nepal and Bangladesh, comparing yield, feasibility and costs. PLoS Negl Trop Dis. 2011;5(2):e960.Joshi AB, Das ML, Akhter S, Chowdhury R, Mondal D, Kumar V, et al. Chemical and environmental vector control as a contribution to the elimination of visceral leishmaniasis on the Indian subcontinent: cluster randomized controlled trials in Bangladesh, India and Nepal. BMC Med. 2009;7:54.S1 AppendixAbbreviations: TDR, Special Programme for Research and Training in Tropical Diseases; WHO, World Health Organization.(DOCX)Click here for additional data file."} +{"text": "Theory of Self- vs. Externally-Regulated Learning\u2122 (SRL vs. ERL) proposed different types of relationships among levels of variables in Personal Self-Regulation (PSR) and Regulatory Teaching (RT) to predict the meta-cognitive, meta-motivational and -emotional variables of learning, and of Academic Achievement in Higher Education. The aim of this investigation was empirical in order to validate the model of the combined effect of low-medium-high levels in PSR and RT on the dependent variables. For the analysis of combinations, a selected sample of 544 undergraduate students from two Spanish universities was used. Data collection was obtained from validated instruments, in Spanish versions. Using an ex-post-facto design, different Univariate and Multivariate Analyses were conducted. Results provide evidence for a consistent effect of low-medium-high levels of PSR and of RT, thus giving significant partial confirmation of the proposed rational model. As predicted, (1) the levels of PSR and positively and significantly effected the levels of learning approaches, resilience, engagement, academic confidence, test anxiety, and procedural and attitudinal academic achievement; (2) the most favorable type of interaction was a high level of PSR with a high level RT process. The limitations and implications of these results in the design of effective teaching are analyzed, to improve university teaching-learning processes.The Educational Psychology. In particular, considerable advances have been made in the knowledge of the roles that metacognitive, meta-motivational, and -affective processes play in university students and Regulatory Teaching (Process of Teaching) to predict cognitive-emotional learning variables (Process of Learning), and achievement (Product of learning) in higher education, offering recent empirical evidence . When the student possesses low personal self-regulation (presage) and is exposed to a low level of regulatory teaching, he/she will present a low level of deep approach with positive emotionality, such as resilience, engagement, and confidence (process), and a high level of negative emotionality, such as test anxiety, ultimately achieving low levels of performance (product).Type 2 combination . When the student possesses low personal self-regulation (presage) and is exposed to highly regulatory teaching, he/she will display a low/moderate level of deep approach with a low/moderate level of positive emotionality, such as resilience, engagement, and confidence, and a moderate/high level of negative emotionality, such as test anxiety (process), ultimately attaining a moderate-low level of performance (product).Type 3 combination . When the student possesses high personal self-regulation (presage) and is exposed to a low level of regulatory teaching, he/she will carry out a moderate level of deep approach with a moderate/high level of positive emotionality, such as resilience, engagement, and confidence, and a low/moderate level of negative emotionality, such as test anxiety (process), ultimately achieving a moderate-high level of performance (product).Type 4 combination . When the student possesses high personal self-regulation (presage) and is exposed to highly regulatory teaching, he will present an \u201cextremely\u201d deep approach with a high level of positive emotionality, such as resilience, engagement, and confidence, and a low level of negative emotionality, such as test anxiety (process), ultimately reaching a high level of performance (product).general meta-ability (like meta-behavior) to control our own thoughts, emotions, and actions. Brown to control our own thoughts, emotions, and actions in learning activity. In essence, this construct adopts the self-regulation postulates of Zimmerman and the other is more general (a deep approach seen as a motivation to understand).Biggs defined resilience and engagement with the task predict students' academic performance, as well as their interest, goals, learning strategies used, effort in study, and academic self-regulated learning the object of their focus (activity-focus); (2) their valence (positive-negative); and (3) degree of activation and emotionality (affective type). In the case of test anxiety, the transactional academic performance . The Biggs's proposal based academic performance on a compendium of types of academic achievement: conceptual (grades achieved on exams), procedural (class attendance and lab work), and attitudinal (class participation and voluntary efforts). Academic performance has taken on greater importance in educational research in recent decades, with many variables being studied in order to identify their influence on the academic performance of university students.Every teaching-learning process aims toward a certain product, with certain objectives and purposes that are meant to result in the student learning some specific subject matter. This product is called effective teaching is a strong predictor of self-regulated learning activities , thus leading to a deep learning approach , as opposed to a surface approach low-medium-high levels of Personal Self-Regulation, as a student variable (as presage variable of learning) will have a statistically significant main effect to level of dependent variables; (2) low-medium-high levels of Regulatory Teaching (process variable of teaching) will have a statistically significant main effect to level of dependent variables; (3) low-medium-high levels of Personal Self-Regulation, in combination with low-medium-high levels of Regulatory Teaching (process variable of teaching), will jointly determine low-medium-high levels of the meta-cognitive variable (learning approach), meta-motivational variables (resilience and engagement), and meta-affective variables in students (process variable of learning), and in types of achievement learning in university students (product variable).The Personal Self-Regulation (PSR), and Regulatory Teaching (RT) we used a total sample of 544 undergraduate students from two universities in the south of Spain. For the analysis of combined relations a selected sample of 201, and 173 students for the type of combination analysis was used. The sample was composed of students enrolled in Psychology, Primary Education, and Educational Psychology degree programs; 86.5% were women and 13.5% were men. Their ages ranged from 19 to 49, with a mean age of 23.08 (\u03c3X = 4.4) years.For the interdependence relations among low-medium-high levels of Personal self-regulation This variable was measured using the Short Self-Regulation Questionnaire (SSRQ) and for the factors of goal setting-planning (\u03b1 = 0.79), decision making (\u03b1 = 0.72) and learning from mistakes (\u03b1 = 0.72). However, the perseverance factor (\u03b1 = 0.63) showed low internal consistency. Correlations have been studied between each item and its factor total, among the factors, and between each factor and the complete questionnaire, with good results for all, except for the decision making factor, which had a lower correlation with other factors (range: 0.41\u20130.58). The correlations between the original version and the complete version, and between the original and the short versions with a Spanish sample (complete SRQ with 32 items and short SRQ with 17 items) are better for the short version than for the complete version .Learning approach (meta-cognitive variable) This was measured with the Revised Two-Factor Study Process Questionnaire, R-SPQ-2F which also yielded acceptable reliability coefficients , similar to those encountered in the study by the original authors, with the AMOS Program. Values for the CFI and NFI range from 0 (poor fit) to 1 Resilience was assessed using the CD-RISC Scale Engagement was assessed with a validated Spanish version of the Utrecht Work Engagement Scale for Students The Academic Behavioral Confidence Scale, ABC ; (2) Procedural scores: these assessed from the student's practical work covering procedural content and skills (four points); (3) Attitudinal scores: these were scores given for class participation and for optional assignments undertaken for a better understanding of the material (two points). In order to carry out the different analyses and compare the results, the different subcompetency scores were converted to an equivalent scale from 1 to 10.df = 48, p < 0.001, CF1 = 0.838, TLI = 0.839, NFI = 0.850, NNFI = 0.867; RMSEA = 0.068) and adequate internal consistency . The ATLP is a self-report instrument to be completed by the teacher and the students, available in Spanish and English versions. It also includes a qualitative part where students can make recommendations for improving each of the processes evaluated. As for the instrument's external validity, results are also consistent, since there are different interdependent relationships among perceptions of variables which exist in an academic environment.Regulatory Teaching . The Scales for Assessment of the Teaching-Learning Process, ATLP, student version .Participants completed the scales voluntarily using an online Using an ex-post-facto design, first a 3 K-means cluster analysis was conducted to establish low-medium-high groups in each of the two variables: Personal Self-Regulation (PSR) and Teaching Regulatory (RT). In the case of the PSR variable, formed the centers of the clusters, response ranges being low (1.00\u20133.07), medium (3.08\u20133.69), and high (3.70\u20135.00). In the case of the RT variable, formed the centers of the clusters, response ranges being low (1.00\u20133.46), medium (3.47\u20134.17), and high (4.18\u20135.00).In addition, several ANOVAs and MANOVAs were carried out, both to establish independence between low-medium-high levels of Personal Self-Regulation and the Regulatory Teaching level, and to ascertain the effect of low-medium-high levels of the independent variables, Personal Self-Regulation and Regulatory Teaching, on the dependent variables examined. Also, using a three factorial design (low-medium-high self-regulation levels) \u00d7 3 (low-medium-high levels of regulatory teaching) several ANOVAs and MANOVAs were conducted, taking as independent variable the afore-mentioned levels. Finally, based on the low-high groups in both variables (PSR and RT) the four kinds of combinations were configured, according to the theoretical model proposed was observed on the total score for this variable, with three homogenous subsets. Similarly, this statistically significant main effect appeared for the levels of its components. The statistically significant partial effect was maintained for the following variables: goals, perseverance, decisions, and learning from mistakes. In all these variables three levels of homogenous subsets appeared, determined by the low-medium-high level of the IV IV (low-medium-high levels) appeared on the total score of this variable, as well as for the levels of its components. A statistically significant partial effect was maintained for the specific regulatory teaching variable, regulatory assessment, preparing to learn, general regulatory teaching, and satisfaction with teaching. Moreover, in all cases, three levels of homogenous subsets appeared, determined by the low-medium-high level of the IV Independent Variable IV (low-medium-high levels) was observed on learning approaches levels. The statistically significant partial effect was maintained for both Deep learning, and Surface learning. The combined analysis of the Personal Self-Regulation IV's effect (low-medium-high levels) on the components of learning approaches yielded a statistically significant main effect. The statistically significant partial effect was retained for deep motivation, for deep strategy, for surface motivation and for surface strategy.A statistically significant general main effect of the Regulatory Teaching IV (low-medium-high levels) was observed on the levels of learning approaches. The statistically significant partial effect was maintained less strongly for Deep learning and more strongly for Surface learning. The combined analysis of the effect of Regulatory Teaching (low-medium-high levels) as regards the learning approaches components, showed a statistically significant main effect. The statistically significant partial effect was maintained for deep motivation but not for deep strategy; a statistically significant partial effect was also observed for surface motivation and for surface strategy on the levels of the total Resilience score was noted. The statistically significant main effect continued for its components, with differential effects depending on the factors: tenacity, stress management, perception of control, adaptation to change, but not for spirituality.A statistically significant global main effect of the Regulatory Teaching IV (low-medium-high) on the levels of total Resilience. The statistically significant main effect was maintained for the combination of resilience's factors as well as for its components, although in a differential way: for tenacity, adjustment to change, and for perception of control, but no statistically significant effect for stress management or for spirituality was noted was recorded on the levels of the total score of Engagement. The statistically significant main effect held for its components, with differential effects depending on the factors: vigor, dedication, and absorption. There also appeared a statistically significant global main effect of the Regulatory Teaching IV (low-medium-high) on the levels of total Engagement. The statistically significant main effect held for its components, with differential effects depending on the factors: vigor, dedication, and absorption.A statistically significant global main effect of Personal Self-Regulation (low-medium-high) on levels of Academic Confidence. The statistically significant main effect was maintained for the effect of the Personal Self-Regulation level on the components of Academic Confidence, as well as for the partial effects on each component: grades, study, and attendance.There appeared a statistically significant main global effect of Regulatory Teaching IV (low-medium-high) on Academic Confidence levels was detected. The statistically significant main effect was retained in the global analysis of the effect of the Regulatory Teaching level on its components, but on partial effects only for two of academic confidence's components , but not for others components .In addition, a statistically significant global main effect of the Personal Self-Regulation IV (low-medium-high) emerged for Test Anxiety, or for its components: Worry and Emotionality. However, there did emerge a statistically significant general main effect of Regulatory Teaching (low-medium-high) on the total score of Test Anxiety. The statistically significant main effect held for the Regulatory Teaching level on its components, with only a partial effect on worry.No statistically significant general main effect of the Personal Self-Regulation IV (low-medium-high) on total achievement was recorded, as well as for the combination of types of achievement, with a statistically significant partial effect for the conceptual type, for the procedural type and for the attitudinal type. Moreover, in a complementary way, there emerged a statistically significant main effect of Regulatory Teaching (low-medium-high level) on total academic achievement, on types of academic achievement and in a specific way only on procedural type.A statistically significant global main effect of the RT independent variable (low-medium-high levels) was noted on the total scores of both types of learning approaches, as well as an interactive effect of PSR \u00d7 RT on them. As regards deep learning, the statistically significant partial effect was sustained for both PSR, marginally, and for RT, with stronger effect.A statistically significant main effect of the PRS levels on learning approaches components emerged. However, a statistically significant main effect of RT (low-medium-high levels) did emerge on learning approaches. The statistically significant partial effect remained for deep motivation, for deep strategy, but not for surface motivation or surface strategy on levels in total Confidence appeared, as well as a partial effect only on the confidence in obtaining grades component.A main effect, though less statistically significant, of the levels in PSR or of levels of RT on total test anxiety, but there was a statistically significant partial effect of level of PS, as well as of level of RT on the worry component.There was no statistically significant main effect of PRS and of TR. Group 1 presented a statistically significant low level in PRS and in TR; group 2 had a statistically significant low level in PRS and a statistically significant high level in TR; group 3 displayed a statistically significant high level in PRS and a statistically significant low level in TR; group 4 had a statistically significant high level in PRS and a statistically significant high level in TR. See Table The univariate (ANOVA) and multivariate analyses (MANOVAs) showed a statistically significant main effect of the four typology students and surface motivation /strategy (1 > 4).There did not emerge any statistically significant main effect of the four type of combination was observed on the level of total resilience (4 > 1). This tendency was observed in the analysis of the effect of type of interaction, as well as of its components, revealing statistically significant effects consistent with tenacity, stress management, and control (4 > 1).Regarding this variable, a statistically significant effect of type of combination on the level of total engagement was noted. Similarly, an effect of type of combination on engagement's components was seen, with consistent statistically significant effects for dedication and absorption .A similar effect of type of combination on the level of total confidence , as well on its components was discerned, with consistent statistically significant effects for confidence in grades and in study .A statistically significant main effect of type de combination on the level of test anxiety was observed, and was especially consistent on the worry component .A statistically significant main effect of type of combination on the level of procedural performance (4 > 1) and attitudinal performance (4 > 1). Direct mean values and statistical effects are presented in Table There appeared a statistically significant main effect of formance > 1 and Self- vs. Externally-Regulated Learning Model does not have any effect on the Regulatory Teaching variable (process teaching variable). That is to say, each of the two variables has a potential explanatory effect on its own and they are not interdependent one on the other.The results of our investigation globally support the various theoretical assumptions proposed thanks to the empirical data we found as regards the predictions of the hypothesis, is that each one of these two variables individually affects learning approach as dependent variables of an established metacognitive kind, although differentially. In this way, while the students' deep approach to learning is determined only by the level of self-regulation, the level of superficial approach is determined both by the lack of self-regulation and by the lack of regulatory teaching as far as superficial motivation and superficial strategy are concerned. This result is in line with others found in previous studies, and clearly establishes the effect of both variables on deep learning approach , on engagement (with regard to dedication and absorption), on academic confidence (to attain grades and study), on lack of anxiety (low level of worry), as well as on performance . These combined effects lend empirical support to the idea that personal and contextual factors interact in the university teaching-learning process , unlike combination 1 (low self-regulation level and low regulatory teaching level), promotes a lower level of superficial approach (and a higher level of deep approach), as well as higher levels of resilience , higher levels of engagement (dedication and absorption) and of academic confidence , lower levels of worry, and ultimately, higher levels of performance. These findings are in the same vein as reports from previous investigations, involving different samples in students, and (2) the role of levels of regulatory teaching (RT), in explaining meta-cognitive, meta-motivational and -affective levels of university students, and their level of procedural and attitudinal achievement in learning. The levels of the PSR and RT reflected significantly positive and interdependent levels of deep learning approach, resilience, engagement, academic confidence, worry, and procedural and attitudinal academic achievement. However, the results offer partial evidence for a consistent four-fold combination typology, thus giving statistically significant confirmation of the proposed rational model. As predicted, (1) the most favorable type of combination is high personal self-regulation with a highly regulated teaching process, yielding high resilience, engagement and confidence level, low worry level, and high procedural and attitudinal performance level; (2) the least favorable type of combination is low PSR level in students with a low RT level, giving rise to surface approach, to low resilience, engagement, and confidence, to high worry, and to low procedural and attitudinal performance level.In general, the new empirical evidence bolsters the importance of the combined effect of (1) levels of implications for understanding and assessing university teaching-learning processes. It is necessary to determine which type of combination is occurring before assigning responsibilities to the teacher as against the student. Even so, the most important aspect is that the proposed model enables us to establish a heuristic of evaluation and analysis of a co-responsible nature in university teaching-learning processes. For this reason, partial models, which only focus on the student or on the teacher in an attempt to predict learning, to attribute responsiblities or to determine performance, should give way to conceptual and interactive models, like the one proposed here. In the light of this empirical evidence and of the Theory of Self vs. Externally-Regulation Learning (de la Fuente, These outcomes also have important All subjects gave written informed consent in accordance with the Declaration of Helsinki. The protocol was approved by the \u201cBIOETIC RESEARCH COMMITE\u201d OF UNIVERSITY OF ALMERIA. Human data were collected according to the Code of Practice of the Council of Psychology of Spain and were kept according to the Spanish Data Protection Act.JF: Coordination of R & D Project, Data collect, Final writing, analysis of data. PS: Final writing, analysis of data. JM-V: Review research, Data collect; MV: Data collect. AG and SF: Review research, Analysis of data.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Medicines Travel Award 2017, we received in total 72 applications from all over the world, most of which were of a very high quality. A decision has been reached by four experts from four different continents and by the editor-in-chief of Medicines.For the Medicines, I am pleased to announce the winner of the Medicines Travel Award for 2017. The travel award was granted to Dr. Peggy M.P.C. Bosch, a postdoctoral research associate at Radboud University, Nijmegen, The Netherlands, working on acupuncture in the treatment of schizophrenia, sleep and depression. The award consists of 800 Swiss Francs to attend any academic conference during 2017.As Editor-in-Chief of Science, The Lancet, Annals of Internal Medicine, etc. Her research focus still lies on acupuncture in psychiatry, although she has also co-authored publications on other themes: Parkinson\u2019s disease, dementia, or methodology of acupuncture research. Recent work, together with colleagues from Kyung Hee University and Harvard University, focused on differences in the neural and psychophysical responses to acupuncture between males and females and also on the differences in responses to acupuncture between left and right sided acupuncture stimulations, using the functional Magnetic Resonance Imaging technique. However, since a lack of research funding has always been a problem for her acupuncture research during all those years, with the support of the Medicines Travel Award, she will now be able to present her work at a highly influential conference this year.Dr. Peggy Bosch holds a master\u2019s degree in Clinical Psychology from Radboud University, Nijmegen, The Netherlands. After receiving her master\u2019s degree, she worked as a psychologist in a forensic psychiatric department, in an addiction department, in an outpatient clinic, and in a trauma department, all at LVR-Klinik Bedburg-Hau in Germany. She worked as an unpaid PhD-student, alongside her clinical psychological work, on a project on acupuncture in the treatment of schizophrenia, sleep and depression and received her PhD-degree from Donders Institute for Brain, Cognition, and Behaviour, Centre for Cognition, Radboud University Nijmegen in 2015. She is currently continuing her research in this direction (as an unpaid postdoc) at both Radboud University and at LVR-Klinik Bedburg-Hau. Dr. Bosch is a reviewer for several journals, was chair at several conferences, and often functions as (keynote) speaker or lecturer. She cooperates with Kyung Hee University, Seoul, Republic of Korea, as well as with Harvard University, Cambridge MA, USA and has published in high impact factor journals, such as Medicines editorial and publishing teams! Congratulations to Dr. Peggy Bosch on behalf of the jury and the"} +{"text": "The International Trade Administration and the National Bureau of Standards, of the Department of Commerce, sponsored a one-day Conference on Standards and Trade on May 5, 1987. The purpose of the Conference was to identify and explore possible actions by the Government and the private sector to enhance U.S. trade through promoting U.S. standards, and the technology they embody, in foreign countries and international organizations.The Conference was co-chaired by the Honorable Bruce Smart, Under Secretary for International Trade, and Dr. Ernest Ambler, Director of the National Bureau of Standards. The first of three segments was comprised of presentations by officials of the U.S. Government: the Honorable Clarence Brown, Deputy Secretary of Commerce, the Honorable Michael Smith, Deputy U.S. Trade Representative, and Ambassador Diana Lady Dougan, U.S. Coordinator, Bureau of International Communications and Information Policy, U.S. Department of State. They stressed the need to improve the national trade picture, described ongoing efforts by the government under the GATT Agreement on Technical Barriers to Trade (the \u201cStandards Code\u201d), bilateral negotiations on standards-related trade problems, participation in treaty organizations and other government-to-government international committees, and the status of the current Uruguay Round of Multinational Trade Negotiations (MTN) talks. They expressed receptiveness to private sector views and their willingness to apply available resources to strengthen our trade position, especially in the international standards arena.The second group of morning speakers addressed standards-related concerns of the business community. Dr. Peter Bell, Vice President of Corporate Technology, Norton Company, focused on export controls and expressed the desire for a single federal regulatory focus. Dr. Peter Bridenbaugh, Vice President for Research and Development, Alcoa, emphasized the need for U.S. leadership in technology and the development of the best technical standards possible. Mr. C. Scott Kulicke, Chairman and Chief Executive Officer Kulicke and Soffa Industries, Inc., described the activities of the Semiconductor Equipment and Materials Institute (SEMI) and the necessity for rapid development of new standards in a changing technological environment, as well as participation by foreign nationals in standards-developing bodies, both in the United States and elsewhere.de facto international standards, ASTM\u2019s role in international standards committee work, and in cooperating with the standards bodies of other countries. Dr. George S. Wham, Chairman, American National Standards Institute, described the private sector standards system and how ANSI functions as the member body to ISO and IEC.Private sector participation in international standardization was the subject of the third part of the morning session. Dr. Robert Baboian, Chairman of the ASTM Board, echoed and underscored the importance of using the best technical standards available as Three Working Groups met in the early afternoon and submitted reports which may be summarized as follows:Catherine Kachurik, CBEMA staff and Director, X-3 Secretariat. The private sector should seek funding by collections under a budgeted, equitable system or by special assessment. Hosting meetings in the United States would reduce costs to U.S. participants and benefit foreign counterparts through favorable exchange rates. The Group proposed media and educational campaigns to convince corporate executives and managers of the long-term value of participating in international standardization activities and to attract replacements for retiring \u201celder statesmen.\u201d The Government should seek industry opinion, be responsive and reactive to standards needs, and support positions in GATT talks and other negotiations.Gerald Ritterbusch, Caterpillar, Inc. Initiatives to promote acceptance of test data by other countries should be pursued at all levels, including the current Uruguay Round of Multinational Trade Negotiations, bilateral negotiations, and the GATT Standards Code Committee. Voluntary laboratory accreditation programs, self-certification, third-party testing, and witnessed tests in the U.S. could reduce testing costs, with treatments matched to products and markets. An industry program should publicize certification programs, their costs, and significance.Barbara Boykin, Aerospace Industries Association. Industry must \u201cthink internationally\u201d about standards and trade, abandoning nationalistic approaches and adapting to the needs of international markets; strengthen representation in ISO and IEC to encourage adoption of U.S. standards; pursue harmonization efforts with the European Community; promote awareness and availability of U.S. standards to potential users abroad; educate U.S. Government officials overseas about standards; preserve private sector leadership and enhance government-industry cooperation.Texts of presentations, summaries of question-and-answer periods, and reports of the Working Groups will be published in Conference Proceedings, available from the NBS Office of Product Standards Policy in July."} +{"text": "Scientific Reports 10.1038/s41598-017-15974-6, published online 16 November 2017Correction to: In the original HTML version of this Article, Hong-Shik Choi, and not Ik Jae Lee, was listed as the corresponding author. Correspondence and request for materials should be addressed to Dr. Ik Jae Lee at ikjae412@yuhs.ac. This has now been corrected in the HTML version of the Article; the PDF version was correct from the time of publication."} +{"text": "The Sino-Austrian High-Tech Acupuncture Research Network was founded in 2005 and has been growing ever since. The network comprises many partners from China and is highly involved in research and education activities. This report introduces the network\u2019s activities in the year 2016. The network was founded in 2005 by Prof. DDr. Gerhard Litscher from Medical University of Graz and comprises many partners from China see .Within 2016, the Sino-Austrian High-Tech Acupuncture Network has grown very fast. Milestones and publications ,10,11,1212\u201319 March 2016: Spring Seminar-Arlberg, Lech-Arlberg, Austria. Function Therapies. Society for Integrative Dental Medicine, CAM in Dentistry is China\u2019s largest comprehensive research institute combining scientific research, medical treatment and teaching that is directly under the State Administration of TCM . It boasts various disciplines, advanced equipment and great research strength and has under it 17 research institutes, six medical institutions, one graduate school, two branch schools, two pharmaceutical companies and publishing houses of ancient books on Chinese medical science. Besides, it is a founder of 13 kinds of academic journals on Chinese medical science.What is especially worth mentioning is the achievement in artemisinin research, which provided a powerful weapon for humans against malaria and saved hundreds of thousands of lives, making tremendous contributions for human health, and thus CACMS were awarded by Lasker Medical Research Award and in 2015 the Nobel Prize in Medicine (Prof. Tu Youyou).Acupuncture is one of the key disciplines of research of this renowned institution. Medical University of Graz (Prof. Gerhard Litscher) has a close cooperation with China Academy of Chinese Medical Sciences (with Prof. Yu Xiaochun) for more than 10 years, and also some activities with Tianjin University of Traditional Chinese Medicine on the topic of high-tech acupuncture research. Within the last years, more than 60 joint SCI/PubMed-listed publications have been published with CACMS alone. Prof. Litscher from Medical University of Graz is Visiting Professor at the Institute of Acupuncture and Moxibustion at CACMS.25 July 2016: New Book-Heart Rate Variability and Acupuncture, Graz, Austria. Contact and order: gerhard.litscher@medunigraz.at Medical Research Centers and 10 Research Institutions. The university occupies 0.65 kmAcupuncture is one of the key disciplines of research of this renowned university. The Medical University of Graz (Prof. Gerhard Litscher) has a close cooperation with Hubei University of Chinese Medicine (with Prof. Wang Hua and Prof. Liang Fengxia) on the topic of high-tech acupuncture research. Within the last years, joint SCI/PubMed-listed publications have been published. Prof. Litscher from the Medical University of Graz is Visiting Professor at Hubei University of Chinese Medicine and at the Hubei Provincial Collaborative Innovation Center of Preventive Treatment by Acupuncture and Moxibustion (Director: Prof. Wang Hua).5 September 2016: Acupuncture Congress\u2014Experts. Timmendorfer Strand, Germany , Beijing, China (et (EPU) .9\u201320 November: USTB-University of Science and Technology Beijing, China Scholars. University of Veterinary Medicine, Vienna, Austria ( Austria ."} +{"text": "The updated data in this report were compiled from situation reports from the Guinea Interministerial Committee for Response Against the Ebola Virus, the Liberia Ministry of Health and Social Welfare, the Sierra Leone Ministry of Health and Sanitation, and the World Health Organization.CDC is assisting ministries of health and working with other organizations to end the ongoing epidemic of Ebola virus disease (Ebola) in West Africa (2), a total of 23,253 confirmed, probable, and suspected cases of Ebola and 9,380 Ebola-related deaths had been reported as of February 15 from the three West African countries where Ebola virus transmission has been widespread and intense. Total case counts include all suspected, probable, and confirmed cases, which are defined similarly by each country (3). Because of improvements in surveillance, the number of cases reported in recent weeks might overestimate the number of Ebola cases in some areas because nonconfirmed cases are included in the total case counts. Sierra Leone reported the highest number of laboratory-confirmed cases , followed by Liberia and Guinea . During the week ending February 14, a daily average of 11 confirmed cases were reported from Sierra Leone, fewer than one from Liberia, and seven from Guinea. The areas with the highest numbers of confirmed cases reported during January 25\u2013February 14 were the Western Area and Port Loko (Sierra Leone) and Forecariah (Guinea) . Guinea http://www.cdc.gov/vhf/ebola/outbreaks/2014-west-africa/index.html. The most up-to-date infection control and clinical guidelines for the Ebola epidemic in West Africa are available at http://www.cdc.gov/vhf/ebola/hcp/index.html.The latest updates on the ongoing Ebola epidemic in West Africa, including case counts, are available at"} +{"text": "The correct title is: The Expression of BTLA Was Increased and the Expression of HVEM and LIGHT Were Decreased in the T Cells of Patients with Rheumatoid Arthritis. The correct citation is: Yang B, Huang Z, Feng W, Wei W, Zhang J, Liao Y, et al. (2016) The Expression of BTLA Was Increased and the Expression of HVEM and LIGHT Were Decreased in the T Cells of Patients with Rheumatoid Arthritis. PLoS ONE 11(5): e0155345. doi:"} +{"text": "Patients with schizophrenia are at increased risk for developing metabolic syndrome, with an estimated prevalence of approximately 35\u201350% . Treatment with atypical antipsychotic medications have been shown to increase rates of metabolic syndrome, with differences observed among antipsychotic agents, most notably in propensity for weight gain: higher for olanzapine, clozapine, and iloperidone; intermediate for quetiapine, risperidone, and paliperidone; and lower for amisulpride, aripiprazole, asenapine, lurasidone, and ziprasidone . Independent of weight gain, atypical antipsychotics also appear to have direct effects on lipid metabolism and glucose regulation. The aim of this safety analysis was to assess the effects of treatment with lurasidone on metabolic syndrome risk in patients with schizophrenia.Changes in the rate of metabolic syndrome during treatment with lurasidone (40\u2013160 mg/d) versus active comparators were analyzed using pooled short-term data from 3 randomized, double-blind, placebo-controlled studies; long-term data from 2 active-controlled studies; and switch data from 2 open-label extension studies. Metabolic syndrome was defined based on the National Cholesterol Education Program criteria .In short-term studies, risk of treatment-emergent metabolic syndrome was similar for patients in the lurasidone and placebo groups ; and was significantly greater for patients in the olanzapine and quetiapine groups compared to placebo. In long-term studies, risk of treatment-emergent metabolic syndrome after 12 months was significantly lower for lurasidone compared with risperidone and non-significantly lower for lurasidone compared with quetiapine XR . In open-label switch studies, the rate of metabolic syndrome decreased in patients switched to lurasidone after 6 weeks of treatment with olanzapine or 12 months of treatment with risperidone.In this comprehensive analysis of the lurasidone clinical trial data base, treatment with lurasidone (40\u2013160 mg/d) was not associated with the development of metabolic syndrome in patients with schizophrenia. Rates of metabolic syndrome increased in patients treated with olanzapine, risperidone, and quetiapine XR."} +{"text": "Practising target-shooting sport requires focused attention and motoric steadiness. A previous non-controlled pilot study suggests that children with impairing symptoms of attention-deficit/hyperactivity disorder (ADHD) benefit from participating in target-shooting sport in local shooting associations, as rated by parents and teachers.This study aims at examining if, and to which extent, target-shooting sport reduces parent- and teacher-reported severity of inattentiveness, hyperactivity, and impulsivity in children with attention difficulties, and if, and to which extend, target-shooting sport improves the children\u2019s wellbeing and quality of life.A mixed method approach is applied. A non-blinded, waiting list controlled study is combined with a case study, consisting of interviews and observations. The intervention consists of children practising target-shooting sport, by attending a local shooting association, once a week for six months, during regular school hours. Data from questionnaires , as well as a computerized continued performance test (Qb test), measure the children\u2019s activity and attention. The study includes 50 children in an intervention group and 50 children in a waiting list control group. The Qb test collects data from at least 20 children from the intervention group and at least 20 children from the waiting list control group. Data from the questionnaires and Qb-test is collected at baseline, and six months post intervention. In addition, a case study is carried out, consisting of interviews of at least five children from the intervention group, their parents, teachers and shooting instructors. Observations are carried out, when children are in school and while they are attending the local shooting association. The case study adds to an in-depth understanding of children\u2019s participation in target-shooting sports.At present, little is known about the effects and influence of practising target-shooting sport for children experiencing difficulties with inattentiveness, hyperactivity and impulsivity. This study is expected to contribute to an understanding of the influence of participating in target-shooting sports on inattentive, hyperactive and impulsive symptoms, and the effects on the children\u2019s psychological wellbeing and quality of life.NCT02898532. Retrospectively registered 14 September 2016.Current Controlled Trials Children diagnosed with attention-deficit/hyperactivity disorder (ADHD) DSM-5) are characterized by three core symptoms: inattention, hyperactivity and impulsivity . Approxi are charTarget-shooting sport may be regarded as mental training, where the participant uses breathing techniques to calm down and focus, and thereby improves attention. Mind and body must be in complete balance during practice. These techniques are somewhat similar to those used in meditation . The DanA mixed method approach is used in a pre-post design Fig. 1)1) combinThe study will investigate beneficial and adverse effects of the intervention on children\u2019s:Inattention, hyperactivity, and impulsiveness.Comorbid symptoms, such as emotional, social and behavioural difficulties.Well-being and quality of life.We hypothesise, that participation in the target-shooting sports in local Danish shooting associations, will reduce the children\u2019s symptoms of inattention, hyperactivity and impulsivity, their comorbid symptoms, and enhance their wellbeing and self-reported quality of life.Inclusion: Children aged 10\u201314, meeting the following criteria: either a clinical diagnosis of ADHD or rated by school staff or school psychologist to have impairing difficulties with inattention, hyperactivity and impulsivity, affecting the child\u2019s school attendance. Included schools are either, schools for children with special needs or municipal schools with special educational programmes for children diagnosed with either ADHD or severe and impairing difficulties of hyperactivity, inattention and impulsivity. If the child receives pharmacological treatment for ADHD, the treatment should not be changed , during the study period.Exclusion: Children with visual impairment, current psychotic symptoms, suicidal ideations or behaviour are excluded from the study.ADHD-RS [ADHD-RS : The queADHD-RS . In thisSDQ - Strengths and Difficulties Questionnaire [ionnaire : The queionnaire .KIDSCREEN-27 [CREEN-27 : The selQb Test [Continued Performance Test, which measures the child\u2019s attention and impulse control. The Qb Test produces quantitative measurements of difficulties in the three areas, characterizing the core symptoms of ADHD; inattention, hyperactivity and impulsiveness. The test has previously been proven valid and sensitive, in the monitoring of the effect of pharmacological treatment of children and young people\u2019s ADHD core symptoms [Qb Test : The Qb symptoms , 24.Semi-structured interviews [terviews , 26: TheObservations [rvations : ObservaThe total score on the 18 items on symptoms of inattention, hyperactivity and impulsivity on the teacher-rated ADHD-RS-IV.The primary outcome is:Hyperactivity (distance and area) and inattention (reaction time variance and omission errors) as measured by the Qb Test.The total score on the relevant 18 items on symptoms of inattention, hyperactivity and impulsivity on the parent-rated ADHD-RS-IV.The total scores on the teacher- and parent-rated SDQ.Quality of life as measured by the total score on the child-rated Kidscreen-27.Secondary outcomes are:Interviews and observations will add to a phenomenological understanding of children\u2019s participation and engagement in their daily life and will qualify to an in-depth understanding of the mechanisms of change.The intervention is organised in collaboration with DGI Shooting, local schools, local shooting associations and the University of Southern Denmark. The intervention is carried out nationwide in Denmark. The children attend the shooting range for about 1\u00a0h, practicing rifle shooting, once a week, for a period of six months. Teachers accompany the children to the local shooting associations, where their shooting instructors meet them. Before the shooting session there is formal introduction and preparation. Every child shoots approximately 20 to 30\u00a0min. As part of each training session, participating children meet up in the club\u2019s common room after the shooting session, where the children receive feedback before going back to school, with their teachers.Due to regulations in the Danish Shooting Associations, riffles are kept in a locked weapons storage cabinet, which requires two separate keys to open. Only members of the Shooting Association, who hold permission from the police, hold the keys for the weapons cabinet. The riffles are never allowed to leave the club area, and throughout the shooting practice an individual, responsible adult instructor will monitor and watch each child, at all times.The participating schools identify, which children are eligible to enrol in the study. Participating children are not randomized, but included either to an intervention group or to a waiting list control group, as they are identified. Waiting list control children are recruited from schools that agree to participate in the study, but will not be part of the intervention until after study period. The children in the waiting list control group have the same characteristics as the children in the intervention group, but are not practicing target-shooting sport during the study period. Parents of all eligible children receive verbal and written information about the intervention, following the guidelines of the National Committee on Health Research Ethics. Upon parental consent, the teacher informs and provides written information to the child. The written information will be read out loud and explained to the child. Background information are provided by parents one week prior to the intervention, three months into the intervention and 6\u00a0months post intervention. ADHD-RS-IV, SDQ, and KIDSCREEN-27 questionnaires are collected one week prior to the intervention. Data, from the waiting list control group, will be collected at the same time. Questionnaires will be distributed to parents and teachers. The children will fill out the questionnaire (Kidscreen-27) and complete the computer test (Qb Test) during regular school hours. A research assistant will carry out the Qb Test at the participating schools.The multiple case study, combining interviews and observations, is nested in the quantitative survey adding substantial in-depth information. Children, parents, teachers and shooting instructors will be interviewed during the intervention. The interviews will mainly focus on experiences with the intervention and changes in quality of life and wellbeing for the children. Interviews with the children, teachers and parents will be carried out either at the school or at the child\u2019s home address. Interviews with shooting instructors will take place in the local shooting associations. The first author will conduct and record all interviews.http://www.quantitativeskills.com/sisa/calculations/samsize.htm. The primary and secondary outcomes measured for both groups, pre- and post-intervention will be analysed using linear mixed-effects models (random coefficient models and multilevel models). With the mixed effects model approach, all available data will be used and intention-to-treat analyses applied. A thematic analysis [Sample size calculations are based on the following parameters: The total score of the first 18 items of ADHD-RS-IV is the primary outcome measurement. In a validation study of a Danish clinical sample of 10\u201317-year-old boys with ADHD, the mean total-score was 32.3 (standard deviation (SD) 10.2) . With ananalysis will be Adapting the environment to provide a clear and concise structure, in regards to routines, time and space, is recognized as a helpful educational and method for children with ADHD symptoms , 29. DevAll these above-mentioned elements are present, when practicing target-shooting sports in local shooting associations in Denmark. There are clearly defined, non-negotiable rules, a clear structure and a predictable setting. This makes target-shooting sport an activity, which is easy to understand and practice, also for children with ADHD. At the shooting range, participants are allocated an individual separate cubicle. Children are supervised at all times by a designated adult instructor, who stays in close proximity of the child. During practice the instructor intends to establish a good connection with the individual child, who benefits from support and recognition. When aiming and shooting, participants receive immediate feedback, as to whether they hit bulls-eye. Preliminary findings from the initial sport project indicate, that participating children enjoys target-shooting sport, show on-going motivation, while at the same time increasing their ability to stay focussed on hitting the target . The uniAt present, no research has investigated the effects of target shooting sports in children with impairing symptoms of inattentiveness, hyperactivity and impulsivity. If found effective in this open trial, larger randomized controlled trials of this intervention are needed. If such trials provide evidence that practising target-shooting sports has beneficial effects, it may become a relevant supplement to the current interventions available for children with ADHD. This may aid the development and adaption of other leisure activities and physical activities and increase the possibilities of participation for children with ADHD. The results may further add to the continued search for effective and motivating non-pharmacological interventions that may help and support children with ADHD."} +{"text": "The following information is missing from the Funding section:The National Natural Science Foundation of China: grants 31471516 and 31271747 to Qingshan Chen; grant 31401465 to Hongwei Jiang; grant 31400074 to Dawei Xin; and grant 31501332 to Chunyan Liu. New Century Excellent Talent Training Plan of Heilongjiang Province Ordinary Institutions of Higher Learning, grant 1252-NCET-004, to Qingshan Chen. Natural Science Foundation Key Program of Heilongjiang Province of China, grant ZD201213, to Qingshan Chen. Qingniancaijun project of Northeast Agricultural University, grant 518062, to Zhaoming Qi. New Century Excellent Talents in University of Ministry of Education, grant NECT-1207-01, to Qingshan Chen. Key Technologies Research and Development Program of China during the Twelfth Five-year Plan Period, grant 2011BAD35B06-1, to Qingshan Chen. Modern Agricultural Industry Technology System, grant CARS-04-02A, to Qingshan Chen. China Postdoctoral Science Foundation, to Zhaoming Qi. Colleges and universities in Heilongjiang Province of the Cheung Kong Scholars backup support program, grant 2014CJHB004, to Qingshan Chen. Harbin good foundation for leaders of disciplines, grant 2014RFXXJ012, to Qingshan Chen. SIPT Project of Northeast Agricultural University: grant 201610224146 to Zhongqiu Ni; grant 201410224111 to Leng Yue; and grant 201510224016 to Shanshan Jiang.The publisher apologizes for the errors."} +{"text": "Launched in 2010, AccessPhysiotherapy is a multimedia online educational resource targeted to physical therapy students, educators, and practitioners [The Advisory Board for AccessPhysiotherapy consists of Annie Burke-Doe of the University of St. Augustine for Health Science in San Diego, California; Eric Shamus, associate professor in the Florida Gulf Coast University Doctor of Physical Therapy program; and Mark Dutton, a physical therapist with Allegheny General Hospital, West Penn Health System, in Pittsburgh, Pennsylvania. Besides their advisory role, the editorial board members have contributed substantial amounts of content to AccessPhysiotherapy and are all authors of well-known physical therapy texts.AccessPhysiotherapy was easy to set up for in-network and remote institutional use via Internet protocol (IP) authentication at the author\u2019s university library. In addition to IP authentication, access via EZProxy, Athens authentication, referral uniform resource locator (URL), and/or username and password is available. The site works well on Windows, MacOS, and Android operating systems and with a variety of browsers. The display automatically resizes to fit mobile device screens . Users cLinks to printable color flyers, brochures, and video tutorials are provided by McGraw-Hill at the time of purchase. COUNTER-compliant statistical reports are available, clearly labeled, and easy to read.AccessPhysiotherapy features a clean, colorful home page with a row of tabs grouping content into broad categories at the top of the page: Home, Books, Quick Reference, Drugs, Multimedia, Cases, Study Tools, and Custom Curriculum . Also frThe basic/advanced search box appearing near the top of the screen allows Boolean and phrase searching. Clicking an arrow displays a menu option that allows federated searching of all Access sites. Results are displayed in context by resource title and can be filtered by type of resource, book title, or topic. The search can be expanded to other AccessMedicine and other McGraw-Hill Medical sites. Interestingly, preliminary statistics at the author\u2019s institution show little use of the search box. Overwhelmingly, users access individual book titles and other resources directly.The Books tab displays available books by title and subject. A core feature of AccessPhysiotherapy is its searchable online collection of McGraw-Hill physical therapy text-books and compilations of case studies. Books with related content in orthopedic examination, pathology, biostatistics, pathophysiology, imaging, and other subjects pertinent to physical therapy are also included. The selection of books has been expanded and currently includes thirty-nine McGraw-Hill titles. Faculty may use the collection to supplement or replace textbooks used in a physical therapy program to guarantee easy, portable online access to course textbooks for all students. MARC 21 records are provided for books included in AccessPhysiotherapy so that records can be included in the library catalog. OpenURL links to PubMed records have been added to many references. A book list can be downloaded in Excel format.When a textbook is selected, the contents are displayed in expandable sections. Textbook chapters may be printed, emailed, or searched by keyword. Full-text textbook content is not downloadable. American Medical Association (AMA)\u2013 and Modern Language Association (MLA)\u2013style citations can be displayed or downloaded. Images and tables that have been approved for educational use may be bookmarked or downloaded to PowerPoint slides. The screen display is clean and easy to navigate. Clickable links to related content and online updates are displayed on the right-hand side of the page.A useful Quick Answers section, available under Quick Reference, provides detailed point-of-care information on conditions of interest to physical therapists. It includes items such as the International Classification of Diseases, Ninth Revision (ICD-9-CM), and ICD-10-CM codes; links to material on the American Physical Therapy Association website; and information on diagnosis, therapy, prognosis, tests, goals, and references with live links to pertinent materials in AccessPhysiotherapy. The \u201cOutcome Measures Toolbox\u201d links to a variety of scales and measures used to assess physical therapy outcomes, with live links to other AccessPhysiotherapy resources. Calculators allow users to compute body mass index (BMI) and other common measurements.A Multimedia section includes Anatomy and Physiology Revealed, a sophisticated interactive cadaver tool from the University of Toledo. Students can peel away layers of realistic-looking virtual cadaver organs to study human anatomy. This feature also includes animations and videos illustrating body functions and disease processes. Other multimedia content, which is updated periodically, includes videos, interactive modules, and lectures. Multimedia content comes from McGraw-Hill products, university physical therapy departments, and other sources.The Study Tools feature can be used to generate custom multiple-choice practice quizzes for the National Physical Therapy Examination. Tests and quizzes can also be generated and scored for classroom use or self-assessment. Students can sign in through MyAccess and review their progress through a feature called My Review Questions. This targeted test review function is a key value-added feature of AccessPhysiotherapy. The Custom Curriculum feature, also accessible via MyAccess, can be used for assignments and course management. If another course management system is already in use, this feature may be redundant.F.A. Davis PT Collection provides additional text-books, cases, and videos. This collection currently includes twenty-nine additional titles and is available at an additional cost. Many Davis titles are widely available through other vendors, so libraries may wish to comparison shop.The AccessPhysiotherapy is an attractively packaged niche product with little direct competition. The most valuable component is the full-text McGraw-Hill physiotherapy e-book collection, which is not widely available else-where. Test preparation is another valuable function of AccessPhysiotherapy that has enjoyed good usage by our students. AccessPhysiotherapy\u2019s online anatomy tool and other multi-media content also add value. Furthermore, for institutions subscribing to AccessMedicine and other McGraw-Hill products, the ability to search multiple products simultaneously with AccessPhysiotherapy is an advantage.McGraw-Hill Education eBook Library.Current institutional pricing for AccessPhysiotherapy is a good value for the amount of information that it provides. The major drawback is that e-book content is limited to McGraw-Hill and Davis products and, therefore, does not provide comprehensive coverage. Indeed, only one of the 2016 Doody\u2019s Core Titles recommended in the \u201cPhysical Therapy\u201d section appears in AccessPhysiotherapy . The e-bGiven this strong caveat, AccessPhysiotherapy merits a thirty-day trial for institutions with a doctoral-level physical therapy program or other academic and clinical physical therapy programs with a need for one-stop-shop physical therapy information. If faculty like and use a good portion of the material, the easy-to-read online textbook display, study tools, and McGraw-Hill e-book collection that AccessPhysiotherapy comprises are worth the relatively moderate subscription price. Its compatibility and inter-face with AccessMedicine and other McGraw-Hill products should increase its chances of surviving in the increasingly competitive e-book market. In summary, AccessPhysiotherapy is a well-designed, easy-to-use product based on McGraw-Hill physical therapy and related textbook content, with some audiovisual and other content add-ins. It offers an appealing, portable platform for physiotherapy students to study textbooks, view videos, and prepare for exams."} +{"text": "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Medical Research Scotland (LP) is incorrectly named in the Funding section. The correct funding information is as follows: Financial support was provided by Carl Tryggers foundation (CTS13-514) ("} +{"text": "Studying de-implementation\u2014defined herein as reducing or stopping the use of a health service or practice provided to patients by healthcare practitioners and systems\u2014has gained traction in recent years. De-implementing ineffective, unproven, harmful, overused, inappropriate, and/or low-value health services and practices is important for mitigating patient harm, improving processes of care, and reducing healthcare costs. A better understanding of the state-of-the-science is needed to guide future objectives and funding initiatives. To this end, we characterized de-implementation research grants funded by the United States (US) National Institutes of Health (NIH) and the Agency for Healthcare Research and Quality (AHRQ).We used systematic methods to search, identify, and describe de-implementation research grants funded across all 27 NIH Institutes and Centers (ICs) and AHRQ from fiscal year 2000 through 2017. Eleven key terms and three funding opportunity announcements were used to search for research grants in the NIH Query, View and Report (QVR) system. Two coders identified eligible grants based on inclusion/exclusion criteria. A codebook was developed, pilot tested, and revised before coding the full grant applications of the final sample.n\u2009=\u200915), with fewer funded by AHRQ, and a majority were funded between fiscal years 2015 and 2016 (n\u2009=\u200911). Grant proposals focused on de-implementing a range of health services and practices across various health areas and delivery settings . Grants proposed to use a variety of study designs and research methods to accomplish study aims.A total of 1277 grants were identified through the QVR system; 542 remained after removing duplicates. After the multistep eligibility assessment and review process, 20 grant applications were coded. Many grants were funded by NIH (Based on the systematic portfolio analysis of NIH- and AHRQ-funded research grants over the past 17\u00a0years, relatively few have focused on studying the de-implementation of ineffective, unproven, harmful, overused, inappropriate, and/or low-value health services and practices provided to patients by healthcare practitioners and systems. Strategies for raising the profile and growing the field of research on de-implementation are discussed. In recent years, as healthcare costs have continued to rise , wastefuhttp://www.preventingoverdiagnosis.net)) and professional society campaigns . Moreov Wisely; \u201318, over Wisely; , 5, 8, i Wisely; , and low Wisely; , 20 is i Wisely; . Such va Wisely; in 2010.Considerations for studying de-implementation, and identification of multilevel, contextual factors that may facilitate or impede de-implementation, have been discussed in the literature. For example, Prasad and Ioannidis described de-implementation processes that may vary as a function of the type of evidence for the practice, including (1) medical practices for which existing evidence is contradictory, (2) medical practices that are unproven, and (3) medical practices that are novel despite widespread use . ImportaTo complement ongoing efforts to study de-implementation, we used systematic methods to analyze grants funded by the US National Institutes of Health (NIH) and the US Agency for Healthcare Research and Quality (AHRQ) using a search database. Consistent with the general goals of portfolio analyses, our objectives were to identify and describe research studies on de-implementation. Such data are critical for assessing the current state-of-the-science, synthesizing findings across health areas and delivery settings, and informing targeted efforts needed to advance research in this area.Consistent with best practices in portfolio analyses, we used the NIH internal-use-only Query, View and Report (QVR) system to identify funded research grants on de-implementation across all 27 NIH Institutes and Centers (ICs) and AHRQ. The analysis included a selective text query involving key search terms along with specific criteria to find the most relevant grants. We limited our search to research-specific grants , including the R-series mechanisms and thechoosing wisely ).Third, consensus meetings on terminology, definitions, measurement, processes, and outcomes are needed to establish a solid foundation for how best to study de-implementation to move this area of inquiry forward. A recognition of the historical roots of studying overuse, underuse, and misuse, including landmark studies and reports e.g., , 52\u201355),\u201355,52\u201355Choosing Wisely campaign [Fourth, better coordination with ongoing de-implementation initiatives and key stakeholders is essential for advancing research on de-implementation. Natural partnerships may include those with the campaign and the campaign , among ocampaign \u201359). Chacampaign , 61\u2014may Several limitations of this study should be noted. Although we used 11 different terms identified from a recent systematic review , relevanFinally, given that our access to full copies of grant applications is limited to two US federally funded research entities , these findings may not generalize to other US-funded entities , Centers for Disease Control and Prevention (CDC)) or non-US research-funding organizations.Over the past 17\u00a0years, relatively few research grants on de-implementation of health services and practices have been funded across NIH and AHRQ. Collaboration is needed among researchers, practitioners, policymakers, patients, and funding agencies to increase the importance of research on de-implementation across health areas, services, practices, and settings. Moving forward, the 20 grants reported herein provide a snapshot of the status of US-funded research on de-implementation and highlight an opportunity for more activity in this area of inquiry."} +{"text": "Nuclear Medicine was established in Myanmar in 1963 by Dr Soe Myint and International Atomic Energy expert Dr R. Hochel at Yangon General Hospital. Nuclear medicine diagnostic and therapeutic services started with Probe Scintillation Detector Systems and rectilinear scanner. In the early stage, many Nuclear Medicine specialists from the International Atomic Energy Agency (IAEA) spent some time in Myanmar and made significant contributions to the development of Nuclear Medicine in our country. The department participated in various IAEA technical cooperation projects and regional cooperation projects. By the late 1990s, new centers were established in Mandalay, Naypyidaw, and North Okkalapa Teaching Hospital of University of Medicine 11, Yangon. The training program related to Nuclear Medicine includes a postgraduate master\u2019s degree (three years) at the University of Medicine. Currently, all centers are equipped with SPECT, SPECT-CT, PET-CT, and cyclotron in Yangon General Hospital.Up until now, the International Atomic Energy Agency has been playing a crucial role in the growth and development of Nuclear Medicine in Myanmar. Our vision is to provide a wide spectrum of nuclear medicine services at a level compatible with the international standards to become a Center of Excellence. MyanmarNuclear Medicine in Myanmar is an inde-pendent medical specialty operated by seven independent departments of nuclear medicine. The country in general and Nuclear Medicine in particular have benefited significantly from the support of the International Atomic Energy Agency (IAEA). IAEA has played and will continue to play a pivotal role not only in successful implementation of the projects, but also in sustainable growth and development of Myanmar Nuclear Medicine.At present, Myanmar has a full range of nuclear medicine facilities including cyclotron and PET-CT. I have experienced the establishment and development of nuclear medicine in Myanmar throughout my lifetime. Thanks to the encouragement and introduction of Asian History of Nuclear Medicine , 3 by PrThe Department of Radioisotope, the forerunner of the future Department of Nuclear Medicine, was founded in 1963 following the visits of the fact-finding mission in 1959, Dr E. H. Belcher, and the then Head of the Division of Life Sciences of IAEA and consultation with the local health authorities in the early 1963 . The fir51Cr-red blood cell survival, ferrokinetic studies with 59Fe, blood volume studies, and vitamin B12 absorption studies into the daily routines was used in lung scintigraphy provided a fully equipped Nuclear Medicine Unit at New General Yangon Hospital in February 1987 . In 1996Research , 5.In January 1998, the Ministry of Health inaugurated a new Department of Nuclear Medicine at Mandalay General Hospital, a teaching hospital affiliated to University of Medicine, Mandalay, the second largest city in Myanmar. In an attempt to provide extended diagnostic services, new departments were established at Thingangyun General Hospital and North Okkalapa Teaching Hospital of University of Medicine II of Yangon in 2002 and 2003, respectively.In January 1999, a Postgraduate Diploma in Nuclear Medicine was inaugurated at the University of Medicine I in Yangon. After eight years, the one-year course was extended to a two-year master program in January 2007. In 2017, the course was extended to three years.In 2009, in vivo and in vitro Diagnostic and Therapeutic Nuclear Medicine was established in a 1000 bedded specialist hospital at Naypyidaw, a new Administrative Capital of Myanmar. There is also a diagnostic Nuclear Medicine department In the 1000-bedded military hospital at Naypyidaw.At present, there are six nuclear medicine centers under the Ministry of Health and one in the private sector. Regarding the equipment, the first dual head SPECT camera was received in 2003, and Nuclear Cardiology imaging was established. In 2014, SPECT-CT cameras were installed in all centers . With thThe department participated in various IAEA technical cooperation projects since 1963, IAEA regional cooperation agreement projects since 1987, and coordinated research projects , 5. NuclMany experts in Nuclear Medicine from other countries have played major roles in the improvement of nuclear medicine services in Myanmar. These experts are Professor E. H. Belcher from Australia, Dr R. Hoschl from Czechoslovakia, Dr L. Kertez from Hungary, Dr R. D. Piyasena from Srilanka, Professor N. Kochupillar from India, Professor Sher M Khan and Dr Iftikhar from Pakistan, Dr Xie Yanfen from IAEA, Professor Felix X. Sundram from Malaysia, Dr Nandrani S. De Zoysa from Srilanka, Mr A. J. M. Bandula Seneveratna from Srilanka, Ms Veronica Wiley from Australia, Professor Brian Hutton from UK, Dr Sai Han from the UK, and Mr Simon Becher, Mr Chatri Hutakari, and Mr Robin Phoon from Thailand and Singapore GE Medical Co.Our vision is to provide a wide spectrum of nuclear medicine services at a level compatible with internationally accepted standards and to become a Center of Excellence in providing efficient, updated, and timely nuclear medicine services."} +{"text": "Nigella sativa (NS) and its isolated compound thymoquinone (TQ) have significant anti-oxidant and anti-inflammatory proprieties, they could represent effective neuroprotective agents. The purpose of this manuscript is to analyze and to recapitulate the results of in vitro and in vivo studies on the potential role of NS/TQ in AD's prevention and treatment. The level of evidence for each included animal study has been assessed by using a modified CAMARADES 10-item checklist. We used MEDLINE and EMBASE databases to screen relevant articles published up to July 2017. A manual search was also performed. The database search yielded 38 studies, of which 18 were included in this manuscript. Results from these approaches suggest that NS or TQ could represent an effective strategy against AD due to the balancing of oxidative processes and the binding to specific intracellular targets. The overall effects mainly regard the prevention of hippocampal pyramidal cell loss and the increased cognitive functions.Several nutraceuticals have been investigated for preventing or retarding the progression of different neurodegenerative diseases, including Alzheimer's disease (AD). Because Although the knowledge of the exact pathophysiological mechanisms remains an unresolved issue, an emerging evidence underlines the role of the oxidative damage and microglia-mediated neuro-inflammatory responses in the initiation of neurodegenerative disorders including Alzheimer's disease (AD) could reduce the risk of AD , also identified as black seed or black cumin, is a flowering plant belonging to the Family Ranunculaceae. In several countries such as the Middle East, Northern Africa and Southwest Asia, NS's seeds and its derivative, have been used not only as a spice and food preservative, but also as a protective and curative remedy for many disorders combined with the clinical condition (\u201cAlzheimer's\u201d OR \u201cdementia\u201d) or the term (\u201cmemory\u201d). Additionally, was performed a manual search of the reference lists of identified articles. Inclusion criteria for study selection were as follows: performed in cell cultures, on animals, systematic reviews and meta-analysis on the topic. All genders and strains were included for animal studies. Published abstracts, summarizing studies, were excluded. Two independent reviewers (V.D.V. and A.B.) screened and assessed for inclusion, the articles identified by the search. The two reviewers agreed on 95% of the selected article and reached consensus on all included studies through collective discussions with a third reviewer (M.R.M.).This review was written according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines 10-item checklist , PC-12, E18, hi-PSC, SH-Y5Y, BV-2, and N2a cells as showed in Table Several in Thymus vulgaris, thymol and carvacrol, and their derivatives had inhibitory effects on acetylcholinesterase (AChE) thanks to their anti-oxidant properties. Thus, indicating that these compounds can be identified as potential therapeutic options for the treatment of AD and/or of cognitive disorders. Ismail et al. and ROS. These data confirmed the hypothesis that TQ, though the mechanism of neuroprotection, may be potentially used in the management of neurodegenerative disorders, including AD. Similar findings were obtained by a study conducted by Alhebshi et al. (S. cerevisiae) (Sxrn1). Interestingly, by using an innovative technology based on the application of Poly Lactic-co-Glycolic Acid (PLGA) nanoparticles, Doolaanea et al. co-encapsulated NSO and plasmid DNA (pDNA) with PLGA nanoparticles in order to enhance gene therapy of AD's syndrome. The authors demonstrated that the encapsulated NSO promoted the outgrowth of murine neuroblastoma cells BV-2, thus resulting in a better efficacy for AD's treatment , in the CA1 region. This effect was correlated with antioxidant activities of TQ on the levels of malondialdehyde, GSH, catalase, and on the superoxide dismutase (SOD) activities. A similar rat model was used by Hosseinzadeh et al. -induced neuroinflammation in rats] it was tested the combination of PNU-120596, a positive allosteric modulator (PAM) of \u03b17 nicotine acetyl choline receptor (nAChR), with the specific nAChR agonist PNU-282987 or with TQ as well as the transition between the M1 phenotype and the alternatively activated phenotypes 2 (M2) which promotes the tissue repairing through the action of anti-inflammatory cytokines and an agent of targeted therapy. For instance, Kitazawa et al. blocked The preclinical research in this field has the difficult task of offering effective therapeutic opportunities to the clinical investigation whereas, to date, the role of nutraceuticals in AD prevention and treatment is of a limited clinical importance. For example, the British Association for Psychopharmacology stated that further evidence is needed to recommend the use of vitamin E and nutritional supplements for this purpose (O'Brien et al., in vitro and in vivo preclinical investigations, and the safety profile of the compounds, should encourage further preclinical investigations and the translation into clinical practice through the drawing of long-term studies conducted on large population size of AD's patients. In particular, further investigations should be performed to elucidate the effectiveness of NS and its individual components. Indeed, despite TQ showed a better nephro-protective effect, no combinatorial studies on NS and its single compounds, have been performed. Moreover, the pharmacokinetics of these compounds, should be elucidated, since data about the brain's bioavailability and transport across blood-brain barrier, are still inconsistent. More pre-clinical studies will be extremely needed in order to highlight the potential interactions of NS/TQ with other drugs, to elaborate the long-term results and, finally to validate and compare the effectiveness of NS/TQ at different stages of AD.To date, there is no drug identifiable as a gold standard in the AD's prevention or treatment. To cover this gap, many neuroprotective agents including nutraceuticals have been evaluated for their potential benefits. This review suggests that NS/TQ could have a significant role for preventing and retarding the progression of AD. The promising results of MC, SB and AB: written the review protocol, scheduled the inclusion and the exclusion criteria and assessed potential articles for inclusion into the manuscript; MM, AV, AF and VD: screened and assessed the articles; AF and AB: independently assessed study quality for the CAMARADES checklist; AB, AC, CA and AA: assisted all the reviewers; AV, AF, GB, CA: revised the tables. AA and GB: revised the English language. All authors partecipate to draft the work, to revise it critically for intellectual contents and approved the final version of manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "There were errors in the Funding statement. The correct funding information is as follows: \"The work done by U.S. Department of Energy Joint Genome Institute is supported by the Office of Science of the U.S. Department of Energy under contract number DE-AC02-05CH11231. This work was supported in part by USDA-ARS CRIS project 3602-22000-015-00D and by Genome Canada and Genome BC LSARPC project 2112. CLS acknowledges support from the Intramural Research Program of the NIH, National Library of Medicine. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\""} +{"text": "There was a mistake in the authorship. The name of Jean-Paul Soulillou was unintentionally omitted. Dr. Soulillou has contributed to collecting and providing important biological samples used for the manuscript, and as such he should be included in the authorship. The authors apologize for the mistake. This error does not change the scientific conclusions of the article in any way.With the inclusion of Dr. Soulillou\u2019s name in the authorship, the paragraph of Author Contributions should also be corrected as follows:SB and CR designed the study; ED, GD, RO, J-PJ, and CP acquired data; MC, PG, MG, and J-PS collected and provided important samples; ED, RD, KG, ND, PP, CP, SC, NG, CR, and SB analyzed and interpreted data; ED, CR, and SB wrote the manuscript; and all authors approved the final version of the manuscript.This correction does not change the scientific conclusions of the article in any way.The authors apologize for the mistake.The original article has been updated.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Drosophila GBA-PD model. Importantly, the authors observed that motor impairments could be rescued by two pharmacological chaperones that reduced ER stress, highlighting a route for potential therapeutic intervention.A recent thorough study by Sanchez-Martinez et al. highlighThe endoplasmic reticulum (ER) is a central organelle for protein folding, lipid synthesis, and calcium storage. Disturbances in ER homeostasis can be caused by multiple factors, including accumulation of misfolded proteins, oxidative stress, and calcium imbalance, resulting in ER stress. This leads to the activation of the unfolded protein response (UPR) triggering a cascade of events aimed at restoring ER homeostasis, or under severe stress activating cell death pathways mutations and triplications , oxidative stress, and lipid metabolism. Together with key pathological PD features such as \u03b1-synuclein aggregation and mitochondrial dysfunction, most of these pathways have been associated with ER stress across different models. For some of these processes however, further work is still required in the specific context of PD, for which we would highlight the emerging role of lipid metabolism and the dynamics of MAM in the interplay between ER stress, mitochondrial function, and autophagy. Finally, since ER stress is associated with aging and known to regulate longevity, further studies on the effect of aging on ER stress in the context PD would be welcomed, as supported by the age-dependent neurodegeneration phenotypes presented by Sanchez-Martinez et al. .To conclude, the work by Sanchez-Martinez et al. supportsHJRF: conceived and wrote the commentary; BJR and RW-M: review and critique. All authors approved it for publication.Work at the Oxford Parkinson's Disease Centre is supported by the Monument Trust Discovery Award from Parkinson's UK (Grant ref: J-1403).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Dear esteemed colleagues and friends,International Brazilian Journal of Urology. We are committed in publishing the highest quality videos detailing novel surgical techniques and approaches by leading surgical teams from across the world. Similarly, we encourage groups pushing the envelope in terms of how we can continually refine the art of surgery and ultimately improve the outcomes of our patients. In this regard, I am pleased to share with you this year\u2019s selection for best videos of the year for 2016. Many individual criteria were taken into account in making this selection including novelty, superior quality in terms of video depiction and narration, and lastly vides that best depict re-defining surgical approaches to urological diseases. On that note, here are the selections:As we begin a new year, I would like to take this opportunity to thank each of you for your commitment in supporting our journal and expanding its readership. This past year has been truly exceptional in the quality of submissions we have received within the video section of the First Prize:Robotic ureteroureterostomy for treatment of a proximal ureteral stricture by Andrade HS et al. from the Glickman Urological and Kidney Institute, Cleveland Clinic published in volume 42(5):1041-1042, September-October 2016 (available at: http://www.intbrazjurol.com.br/video-section/andrade_1041_1042) published in volume 42:168-169,Third Prize:Retzus-sparing robotic-assisted laparoscopic radical prostatectomy: A step-by-step technique description of this first Brazilian experience by Tobias-Machado M et al. from the Departmento de Urologia, Faculdade de Medicina do ABC, Santo Andre, SP, Brazil and other centers of excellence in Sao Paulo, SP, Brazil published in volume 42(6):1250, November-December 2016 (available at: http://www.intbrazjurol.com.br/video-section/tobias-machado_1250_1250) . This suInternational Brazilian Journal of Urology. We are committed in publishing the very best original articles and videos from across the world and similarly will continue to do so in a very timely manner, with a rigorous peer review process by the very best subject leaders. Lastly, I send my very best wishes to you and your families for 2017.I would like to conclude this editorial by once again thanking each of you for the support of the Video Section Editor,International Brazilian of UrologyAssociate member, Department of GU OncologyMoffitt Cancer CenterAssociate Professor, Department of UrologyUniversity of South FloridaTampa, Florida, USAE-mail: philippe.spiess@moffitt.orgPhilippe E Spiess, M.D, MS, FACS, FRCS(C)"} +{"text": "The term \u2018early life stress\u2019 has been used to describe a broad spectrum of adverse exposures during foetal life, childhood and adolescence. Early life stress and trauma are associated with a higher risk for later mental and physical health disorders, such as anxiety, depression, and post-traumatic stress disorder (P.T.S.D.) as well as cardiometabolic and inflammatory diseases and chronic pain syndromes. The objective of this brief review is to investigate the neuroendocrine responses to early life stress and their role as biological predisposing factors for later disease.Stress-related neuroendocrine alterations in response to early adversity include hyper- or hypo-activation of the stress system and may persist or worsen in later life, acting as biological vulnerability factors for the development of later disease. A key effect of stress during foetal life, childhood, and adolescence is that it programmes the developing brain, especially brain structures involved in stress reactions, such as the prefrontal cortex, the hippocampus, and the amygdala, to hyper- or hypo-react to ensuing stressors. Animal studies have shown that chronically elevated stress mediators may lead to alterations in brain development through mechanisms of accelerated loss of neurons, delays in myelination, or abnormalities in developmentally appropriate neural synaptic pruning. Critical periods of brain development represent time-windows of elevated synaptic plasticity, mediating vulnerability, or establishing resilience to stress.Stress is generally associated with acute activation of the hypothalamic-pituitary-adrenal (H.P.A.) axis and the arousal/sympathetic nervous system, as evidenced, in most studies, by elevated cortisol and catecholamine concentrations in the periphery. However, the chronic and/or intense experience of stress may be associated with chronic hyper- or hypo-activation of mediators of the stress system. This chronic condition represents dyshomeostasis, also called allostasis or cacostasis, which is related to further morbidity, such as obesity and the metabolic syndrome, diabetes mellitus type 2, atherosclerosis, osteoporosis, and immune dysfunction was noted in children that continued to exhibit P.T.S.D. This longitudinal interaction of peripheral cortisol and NE concentrations seems to characterize those that develop and maintain P.T.S.D. in P.T.S.D., and among them, previous trauma and time since the trauma occurred seem to be crucial determinants of hyper- or hypo- activation of the H.P.A. axis. Other parameters include: genetic and epigenetic vulnerability; transgenerational actions; environmental influences in the management of early life stress; the nature, severity, and duration of the traumatic event; the developmental stage, age at the time of trauma; and comorbidities (Pervanidou, In conclusion, early life stress exposures result in neuroendocrine alterations and programming of the neuronal networks of the brain, acting as predisposing factors for further mental and physical health problems."} +{"text": "Background: Impairment in orientation to time and place is commonly observed in community-dwelling older individuals. Nevertheless, the clinical significance of this has been not fully explored. In this study, we investigated the link between performance in orientation domains and future risk of cardiovascular events and mortality in a non-hospital setting of the oldest old adults.Methods: We included 528 subjects free of myocardial infarction (Group A), 477 individuals free of stroke/transient ischemic attack (Group B), and 432 subjects free of both myocardial infarction and stroke/transient ischemic attack (Group C) at baseline from the population-based Leiden 85-plus cohort study. Participants were asked to answer five questions related to orientation to time and five questions related to orientation to place. 5-year risks of first-time fatal and non-fatal myocardial infarction, fatal and non-fatal stroke, as well as cardiovascular and non-cardiovascular mortality, were estimated using the multivariate Cox regression analysis.Results: In the multivariable analyses, adjusted for sociodemographic characteristics and cardiovascular risk factors, each point lower performance in \u201corientation to time\u201d was significantly associated with higher risk of first-time myocardial infarction , first-time stroke , cardiovascular mortality and non-cardiovascular mortality . Similarly, each point lower performance in \u201corientation to place\u201d was significantly associated with higher risk of first-time myocardial infarction , first-time stroke , cardiovascular mortality and non-cardiovascular mortality .Conclusions: Lower performance in orientation to time and place in advanced age is independently related to higher risk of myocardial infarction, stroke and mortality. Impaired orientation might be an early sign of covert vascular injuries, putting subjects at greater risk of cardiovascular events and mortality. Current evidence indicates that impaired cognitive function is associated with cardiovascular events and mortality participated in the study. Within a month after their 85th birthday, a physician or a research nurse contacted them by phone to request their participation. If subjects agreed to participate, they were visited at their place of residence. For this study, we excluded three subjects with uncompleted cognitive tests at baseline. Furthermore, based on outcome variable, we excluded subjects with the previous clinical history of myocardial infarction (Group A), clinical stroke and/or transient ischemic attack (TIA) (Group B), and from history of both myocardial infarction and stroke/TIA events (Group C) at baseline. Information obtained from general practitioners or nursing home physicians. In total, data of 528 subjects free of clinical myocardial infarction (Group A) and 477 subjects free of clinical stroke/TIA (Group B) at baseline were analyzed to evaluate the risk of future myocardial infarction and stroke respectively. Additionally, 432 subjects without a history of both myocardial infarction and stroke/TIA events (Group C) at baseline were examined for cumulative risk of cardiovascular events. Moreover, subjects in Group C were further tested for 5-year risks of cardiovascular, non-cardiovascular and all-cause mortality.Data on orientation domains were extracted from standard administrated Mini-Mental State Examination (MMSE) at the time of entry to the study. To evaluate orientation domains, each participant was asked to answer five questions related to orientation to time and five questions related to orientation to place and were received scores ranged from zero to five. Lower scores in each test indicate greater impairment in orientation domains. Risks of cardiovascular events and mortality were assessed continuously per each point lower scores in orientation to time and orientation to place.First fatal and non-fatal myocardial infarction, stroke, and cumulative cardiovascular events were considered as the main outcomes. Moreover, 5-year cardiovascular, non-cardiovascular and all-cause mortality were assessed. Information about health status and events were recorded with general practitioners. The occurrence of clinically recognized myocardial infarction and stroke during 5 years of follow-up was assessed by annually interviewing general practitioners and were recorded according to the International Classification of Disease and Related Disorders, Tenth revision. Death caused by myocardial infarction was classified as ICD-10 codes I21-I22 . All participants were interviewed for the current and past history of smoking. Diabetes mellitus was considered present when included in the records of the primary care physician when non-fasting glucose concentrations were more than 11.0 millimoles per litter, or when a participant was using anti-diabetic medication according to their pharmacy records. Hypertension was defined as the history of hypertension, or use of antihypertensive medication at baseline. History of cardiovascular diseases, including atrial fibrillation, myocardial infarction and stroke/TIA was obtained from general practitioners or nursing home physicians at baseline.Baseline characteristics of subjects reported as mean with standard deviation for continuous variables and frequency with the percentage for categorical variables. We calculated incidence rates by dividing the number of events by person-years at risk during the follow-up period. The incident myocardial infarction and incident stroke and mortality in relation to orientation to time and place were calculated with Cox regression and reported by hazard ratio (HR) with 95% confidence interval (CI) in three models. Firstly, we calculated hazard ratios for the association between orientation domains and outcomes (crude model). In the second model, analyses were adjusted for sex and education (adjusted model 1). Finally, analyses were further adjusted for current smoking, body mass index, total cholesterol, systolic blood pressure, history of hypertension, history of diabetes mellitus, history of atrial fibrillation and history of previous stroke/TIA events in subjects free of myocardial infarction or history of previous myocardial infarction events in subjects free of stroke/TIA (adjusted model 2). Moreover, we tested whether the risk of cardiovascular events was independent of the clinical cut-off points of the MMSE test in impaired (MMSE < 24) vs. preserved (MMSE \u2265 24) level of cognitive function. P for interaction was calculated by adding an interaction term produced by multiplying scores of orientation to time and place and stratified variable. All analyses were conducted using SPSS statistical software .Table P = 0.007) of myocardial infarction in Group A and 1.35-fold higher risk of stroke in Group B. Similarly, one point lower score in orientation to place was associated with 1.67-fold higher risk of myocardial infarction in Group A and about 1.39-fold higher risk of stroke in Group B. Overall, each point lower score in orientation to time in Group C was associated with about 1.39-fold and each point of decline in orientation to place was associated with around 1.52-fold higher chance of cardiovascular events.Table P = 0.009), non-cardiovascular , and all-cause mortality. Similar findings were observed for the association between performance in orientation to place with 5-year cardiovascular , non-cardiovascular , and all-cause mortality.Table P for interaction <0.05).Figure In this population-based study of the oldest old individuals, we present that lower performance in orientation to time and place, independent of socio-demographic and conventional cardiovascular risk factors, is associated with greater risk of incident myocardial infarction in Group A , stroke in Group B (free of stroke/TIA at baseline), and cumulative cardiovascular events in Group C . Moreover, when we examined mortality due to cardiovascular or non-cardiovascular causes in Group C, it is increased in relation to poorer performance in orientation to time and place.The oldest old population is the fast growing segment of the western population (Robine and Paccaud, Several studies showed that orientation to time and place are among the first domains of cognitive function that get affected in elderly individuals and among the earliest domains to be lost in dementia (Fillenbaum et al., Our findings on the association between poor performance in orientation to time and place with higher incident cardiovascular events and mortality are in line with previous reports, demonstrating that impairment in orientation to time and place was associated with higher incident cardiovascular events and lower survival among elderly (O'Donnell et al., Our study has strengths and limitations. As strength, this study included relatively large population of the oldest old individuals with long follow-up time and availability of extensive data on sociodemographic, cardiovascular factors and mortality causes. The first possible limitation is the lack of neuroimaging data to identify subjects with covert brain vascular abnormalities at baseline. As a second possible limitation, this study was conducted in a sample of very old Dutch population, and thus, the results should be confirmed in other geographic, racial or ethnic subjects. The third limitation of the study is the lack of evidence of physical activity and mental health of participants which could potentially influence the results.In conclusion, our findings suggest that lower performance in orientation to time and place, in the oldest old individuals, is associated with incident myocardial infarction, incident stroke and mortality. This evidence might suggest that applying simple, fast and practical tests like orientation to time and place can be further considered for enhancing prediction models for identifying the oldest old subjects who are at the increased risk of cardiovascular events and cerebrovascular accidents. Since previous studies that evaluated the association between cognitive function and future risk of cardiovascular events and mortality were performed mostly in the white middle aged and young elderly populations, the generalizability of those findings is limited. To improve the external validity, different age groups of elderly with diverse ethnic background and lifestyle are needed to be included in the future studies.Study concept and design: BS, SR, and AdC. Data analysis: SR. Data acquisition and interpretation, critical revising of the manuscript, and agreement to be accountable for all aspects of the work: SR, MvB, JJ, JG, RP, AdC, and BS. Drafting of the manuscript: SR and BS. Final approval of the version to be published: SR, MvB, JJ, JG, RP, and BS.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The use of information and communication technologies (ICTs) for information sharing among family members is increasing dramatically. However, little is known about the associated factors and the influence on family well-being.The authors investigated the pattern and social determinants of family life information sharing with family and the associations of different methods of sharing with perceived family health, happiness, and harmony (3Hs) in Hong Kong, where mobile phone ownership and Internet access are among the most prevalent, easiest, and fastest in the world.A territory-wide population-based telephone survey was conducted from January to August 2016 on different methods of family life information sharing with family members, including face-to-face, phone, instant messaging (IM), social media sites, video calls, and email. Family well-being was assessed by three single items on perceived family health, happiness, and harmony, with higher scores indicating better family well-being. Adjusted prevalence ratios were used to assess the associations of sociodemographic factors with family life information sharing, and adjusted beta coefficients for family well-being.P for trend <.01). Higher education was most strongly associated with the use of video calls . Higher household income was significantly associated with the use of any method, face-to-face, and IM . Sharing family life information was associated with a higher level of perceived family well-being , especially by face-to-face and video calls . The combination of face-to-face and video calls was most strongly associated with a higher level of perceived family well-being .Of 2017 respondents, face-to-face was the most common method to share family life information , followed by IM , phone , social media sites , video calls , and email . Younger age and higher education were associated with the use of any (at least one) method, face-to-face, IM, and social media sites for sharing family life information (all The differential use of ICTs to share family life information was observed. The prevalence of video calls was low, but associated with much better family well-being. The results need to be confirmed by prospective and intervention studies to promote the use of video calls to communicate and share information with family, particularly in disadvantaged groups. Family life information refers to information that strengthens family functioning through improving communication skills, knowledge about developmental tasks, decision-making skills, self-esteem, and interpersonal relationships . PreviouWith advances in technology and high prevalence of mobile phone ownership and Internet penetration, the use of information and communication technologies (ICTs), including instant messaging (IM), social media sites, video calls, and email, to share information is increasing dramatically. These newly emerging ICTs enable people to communicate and share information more conveniently, interactively, and at lower cost. For instance, IM enables users to send information by text, photograph, audio clips, and video at any time, and can reach many individuals simultaneously. Social media sites allow for interconnectivity and provide a platform for information sharing. Video calls provide visual cues along with immediate interaction and feedback for geographically separated individuals .Hong Kong is the most modernized and westernized city in China. There is also widespread penetration of mobile phones and the Internet owing to the advanced cyber-infrastructure and low cost of access to the Internet . Mobile However, people with low socioeconomic position (SEP) (lower education or income) often have low access and usage of ICTs ,21, whicDespite the high prevalence of ICT use, traditional communication methods (face-to-face and phone) are most used in a family context in Hong Kong, along with a higher level of family well-being . Face-toTo the best of the authors\u2019 knowledge, no studies have investigated the use of different methods to share family life information or its association with family well-being. The authors used a large population-based telephone survey to investigate the pattern and social determinants of family life information sharing with family and the associations of different methods of family life information sharing, especially video calls, with perceived family well-being in Chinese adults living in Hong Kong. The authors examined whether the findings support the Inverse ICT Law on family information sharing.The Hong Kong Family and Health Information Trends Survey (FHInTs) was part of a project entitled \u201cFAMILY: A Jockey Club Initiative for a Harmonious Society.\u201d FHInTs was a regular periodic population-based telephone survey of the general Hong Kong public\u2019s opinions and behaviors on family health, information use, and health communication. Since 2009, five waves of FHInTs have been conducted and details are reported elsewhere ,26. The All interviews were conducted by trained interviewers of the Public Opinion Program at The University of Hong Kong using a Web-based computer-assisted telephone interview system. The survey targeted the Cantonese-speaking adult population aged 18 years and older. Hong Kong residents aged 18 or older were eligible to participate in the telephone survey. Respondents who were psychologically or physically unable to communicate or were unable to communicate using Cantonese over the phone were excluded. Landline telephone numbers were randomly generated using known prefixes assigned to telecommunication services providers under the Numbering Plan provided by the government Office of the Communications Authority. When contact was successfully established with a target household, one qualified person was selected from all those present using the \u201cnext birthday\u201d rule . The perThe most recent wave consisted of four subsets: health, health information, family information, and family communication. Each subset had core questions and subset-specific questions. The authors set the sampling error at 3.1% with 5% type I error. Based on the population size in mid-2009 , the autDefinitions of family and family life information (as mentioned previously) were explained to the respondents before asking questions about family life information sharing and family 3Hs. Methods of family life information sharing were assessed by asking respondents the usual methods of sharing family life information with their family, including face-to-face, phone, IM, social media sites, video calls, and email. Family 3Hs were measured by using three separate questions with a score from 0 to 10. Family well-being was calculated based on the composite score of the 3Hs with higher scores indicating better family well-being. In this sample, the Cronbach alpha coefficient of family well-being was .89, indicating good internal consistency .Socioeconomic position was measured using educational attainment, employment status, and monthly household income. Educational attainment was categorized as primary or below, secondary, and tertiary or above. Employment status was categorized as full time, part time, self-employed, and unemployed. Monthly household income was categorized as HK $40,000.P value <.05 was considered statistically significant.To improve the representativeness of the findings, the raw data were weighted using the random iterative method ,42 accorOf 2017 respondents after weighting, most were women, aged 25 to 64 years, and married or cohabitating . Most reP<.001).In the total sample after weighting, the most common method of family life information sharing was face-to-face , followed by IM, phone, and social media sites . Only a P<.05) (P for trend <.01). Higher education level was associated with the use of any method, face-to-face, IM, social media sites, video calls, and email , with the strongest association observed for video calls . Higher monthly household income was associated with the use of any method, face-to-face, and IM to share family life information . However, household income was inversely associated with the use of phone, particularly for respondents with household income higher than HK $40,000 .More women shared family life information than men by all methods except face-to-face . YoungerFamily life information sharing by any method and face-to-face were strongly associated with higher levels of perceived family health, happiness, harmony, and overall family well-being . The useCompared with the respondents who had never shared family life information, the use of both face-to-face and video calls appeared to be most strongly associated with higher levels of perceived family health , happiness , harmony , and overall family well-being , although the 95% CIs overlapped with the use of face-to-face only .This study provides the first evidence of family life information sharing in one of the most developed non-Western urban settings with high penetration of mobile phones and Internet, and widespread and fast Internet connection. Although the 95% CIs overlapped, it is noteworthy that respondents with higher education were much more likely to share family life information with family by video calls . This study also found that family life information sharing by the combination of face-to-face and video calls appeared to be most strongly associated with higher levels of perceived family 3Hs and overall well-being.This study shows that women in Hong Kong are more likely to share family life information than men are, especially by ICTs. However, researchers showed an emerging trend that both genders have equal access to the Internet in developing Asian countries such as Vietnam . Given tA previous survey showed that people with low SEP were less likely to seek family life information online and use ICTs to communicate with family members ,28. ThisMoreover, the authors found that family life information sharing was associated with all three dimensions of family well-being . Intervention studies have found that family life education programs have benefits of forming and sustaining healthy relationships and improving family functions because family life information can help manage family activities, cope with family problems effectively, and deliver care of the children and the elderly ,47. In aNotably, the authors found that the use of face-to-face sharing of family life information was most strongly associated with all three dimensions of family well-being. Previous studies in Hong Kong and elsewhere ,49,50 haThis study has some limitations. First, the cross-sectional study could have residual confounding or the temporal sequence of family life information sharing and family well-being was uncertain. Second, the methods of family life information sharing were determined only by a simple yes/no question; more detailed information such as the frequency of using face-to-face or ICTs to share should be collected in future research. Nevertheless, the authors have shown that a simple question could yield preliminary data to show the presence of the Inverse ICT Law and guide more in-depth studies. Third, the sampling method only covered adults. However, adolescents are more active digital users and more likely to embrace ICTs in various forms. Exploration of ICT use in young people may enable a better understanding of how it affects family well-being. In addition, online interpersonal influences may affect health-related quality of life in adolescents . FinallyThis study suggests several avenues for future research. First, qualitative research on family life information sharing should be conducted in this setting for a deeper understanding of information sharing behaviors in a family context. Prospective cohort studies and intervention studies are also needed to assess the impact of family life information sharing by video calls on family well-being. Second, information on specific groups of families is important to further address the impact of ICT use on those with special needs. For instance, in families with members living in geographically separated areas, ICTs such as video calls can be increasingly used to maintain family relationships and bonds . ICTs suAlthough the impact of ICT usage on family has been extensively studied, this study has provided the first evidence of different methods of information sharing with family, especially video calls, and their associations with family well-being. The differential use of ICTs to share family life information supports the emergence of the Inverse ICT Law. Face-to-face communication remained the main mode for family life information sharing and was associated with better family well-being. The prevalence of video calls was low but associated with better perceived family well-being, denoting a feasible way by better use of ICTs to improve family well-being. Further prospective and intervention studies are warranted to confirm the results and to promote the use of video calls to communicate and share information with family, particularly in disadvantaged groups."} +{"text": "A study on age-related electrocardiographic (ECG) changes was conducted on 20 apparently healthy Black Bengal goats with no history of cardiac disorders during 2015-2016.The goats selected for the study belonged to four different age groups; Group 1: Goats up to 6 months of age, Group 2: Above 6 months and below 1 year of age, Group 3: Above 1 year and below 2 years of age, and Group 4: Above 2 years of age. The ECG was recorded with the animals in standing position using a 12-lead standard ECG recorder . The paper speed was set to 25 mm/s with the sensitivity of the machine was adjusted at 1 (1 cm=mV).The ECG parameters were compared within different age groups, and the data were analyzed statistically using SPSS 16.0 taking a significant level of 95% (p<0.05) in all cases. The lead-I ECG revealed a significant difference in amplitude of QRS complex, PR interval, QT interval, RR interval, PQ segment, ST segment, TP segment, and heart rate among some age groups. In bipolar limb lead-II, the amplitude of T-wave, RR interval, ST segment, TP segment, and heart rate was a significant difference among some age groups. Lead-III presented significant difference among age groups in different parameters such as QRS complex duration, T-wave duration, RR interval, ST segment, TP segment, and heart rate.The study concluded that there is a significant variation in the ECG parameters both in terms of values and configuration of ECG waves when age is taken into consideration. The results of the study might be used as a reference value for field veterinarians. An electrocardiogram (ECG) is the recording of electric potentials generated by the cardiac impulse by placing electrodes on the skin on opposite sides of the heart . ResearcConsidering the perspective of clinical treatment, a correct ECG diagnosis can only be done if the ECG values of the screened goats are compared with normal reference values established in healthy goats. The size and shape of the hearts in goats vary according to its breed and body size, and this variation is expected to be reflected in the ECG . The BlaAs per our knowledge, there is no report on ECG change with advancing age in the Black Bengal goat. Therefore, the present study was conducted to assess the alterations in ECG parameters of Black Bengal goats in different age groups which may act as a source of reference for veterinary clinicians while interpreting the ECG.Approval of Animal Ethics Committee is not required in no-invasive types of experiments.The animals were assigned into four groups according to their age, comprising Group 1 , Group 2 , Group 3 , and Group 4 . The animals were examined before ECG recordings. The animals were examined for clinical signs such as coughing, paleness of mucous membrane, fatigue, shortness of breath, and swollen abdomen. The animals showing no such clinical signs and history of cardiac disorders were selected for the study. A 12-lead standard ECG recorder, Cardiart 108 T-MK VII- BPL, India, was used to record ECG in all the goats. ECG was taken with the animal restrained in standing position on a wooden board. The ECG machine was set for a paper speed of 25 mm/s and sensitivity of 1 (1 cm=1 mv) with the 50 cycles filter turned \u201con.\u201d The anterolateral aspect, just below the elbow, and stifle joint were the preferred sites of attachment of crocodile clips, and the ECG tracings were taken in three bipolar standard leads as described by Ahmed and Sanyal [The parameters were compared using MS Excel and SPSS-10 software.The results mentioned here are in accordance to Tables-In this study, the amplitude of QRS complex had no significant difference among the different age groups in lead-II and lead-III, whereas in lead-I, the QRS complex amplitude of Group C animals was significantly lower than the other groups. The configuration of QRS complex varied widely as indicated in The T-wave amplitude of Black Bengal goats belonging to different age groups did not reveal any significant difference between age groups in lead-I and lead-III. However, in lead-II, the amplitude of T-wave varied significantly within the group, and the highest amplitude was measured in Group D. The duration of T-wave belonging to different age groups was found to be statistically insignificant in lead-I. However, in lead-II, the duration of T-wave was significantly higher in Group D, and in lead-III, significantly higher amplitudes were obtained in Groups B , C, and There was no significant difference between PR interval and QT interval in lead-II and lead-III. However, in lead-I, both the PR interval and QT interval were found to be significantly lower in Group A. R-R interval in Black Bengal goat shows no significant difference between lead-I and lead-II, whereas in lead-III, the R-R interval has a significant difference within the group.Group D recorded significantly higher PQ segment in lead-I and lead-II, whereas in lead-III, there was no significant difference within groups. In Black Bengal goat, the ST segment was recorded as significantly different in lead-I, lead-II, and lead-III. There was a significant difference between groups with regard to TP segment of Black Bengal goat in all the leads. Lead-I and lead- II electrocardiogram tracings of Group C and Group D are shown in Figures Significantly higher heart rates were invariably observed in the youngest goats, i.e., Group A in all the leads taken into consideration.et al. [Our observations are not in agreement with Atmaca et al. , who recThere was no statistical significance between the amplitudes of QRS complex in leads II and III. However, the QRS complex amplitude was significantly lower (p<0.05) in Group C of lead-I, as compared to other age groups. The low amplitude QRS complex might be due to high degree of synchronized ventricular depolarization . The varThe amplitude of T-wave of Black Bengal goats belonging to different age groups did not reveal any significant difference between age groups in lead-I and lead-III. However, in lead-II, the amplitudes of T-wave varied significantly within groups, and the highest amplitude was measured in Group D. T-wave amplitude recorded positively in all leads. The present finding is in agreement with Ahmed and Sanyal .et al. [The mean duration of T-wave belonging to different age groups was found to be statistically insignificant in lead-I. However, in lead-II, the duration of T-wave was significantly higher in Group D, and in lead-III, significantly higher values were obtained in Groups B, C, and D in comparison to Group A. Our values are not in agreement with Mohapatra et al. , who repet al. [The PR interval denotes the atrioventricular conduction time. There was no significant difference between PR intervals in lead-II and lead-III, whereas Group A exhibited a significantly lower (p<0.05) PR interval in lead-I. Shorter PR interval results in shorter interval between successive cardiac cycles. Mir et al. also repet al. . R-R intet al. .The PQ segment was found to be significantly higher in older animals (Groups C and D) as compared to younger ones (Groups A and B) in leads I and II, whereas no significant difference was observed amidst groups in lead-III. Analysis of variance revealed that there was a significant difference of ST segment in leads I, II, and III. In lead-I, it was found that younger goats had higher ST segment than older ones, while ST segment in lead-II and lead-III was higher in older animals. The TP segment of Black Bengal goat in different leads differed significantly within age groups.Significantly higher heart rates were invariably observed in the youngest animals, i.e., Group A in all the leads taken into consideration. Higher heart rates in kids might be due to stress and excitation caused by separation of kids from their does .The study concludes that age has direct influence on the outcome of values of ECG parameters as well as, the configuration of different ECG waves varies with respect to age and also within the same age group of goats. So, while analyzing ECG in apparently healthy Black Bengal goats, all these factors should be taken into consideration for proper or an accurate interpretation of cardiac originated problems from the ECG only.The work was carried out, and the manuscript was written by RRP as a part of her M.V.Sc. research program, APKM was the chairman of the advisory committee. RRP, APKM, SM, and AKK designed the work. RRP, SM, and TJ performed the field work of recording ECG. RRP and TJ performed the statistical analysis. SM edited the article. AKK and APKM made the final revision of the manuscript. All authors read and accepted the final manuscript."} +{"text": "This paper presents a brief description of research conducted by the consortium, database, and portal development and upcoming features that will enhance the LungMAP experience for a community of users.The National Heart, Lung, and Blood Institute is funding an effort to create a molecular atlas of the developing lung (LungMAP) to serve as a research resource and public education tool. The lung is a complex organ with lengthy development time driven by interactive gene networks and dynamic cross talk among multiple cell types to control and coordinate lineage specification, cell proliferation, differentiation, migration, morphogenesis, and injury repair. A better understanding of the processes that regulate lung development, particularly alveologenesis, will have a significant impact on survival rates for premature infants born with incomplete lung development and will facilitate lung injury repair and regeneration in adults. A consortium of four research centers, a data coordinating center, and a human tissue repository provides high-quality molecular data of developing human and mouse lungs. LungMAP includes mouse and human data for cross correlation of developmental processes across species. LungMAP is generating foundational data and analysis, creating a web portal for presentation of results and public sharing of data sets, establishing a repository of young human lung tissues obtained through organ donor organizations, and developing a comprehensive lung ontology that incorporates the latest findings of the consortium. The LungMAP website ( Development and maintenance of the structure of the lung requires interactive gene networks and dynamic cross talk among multiple cell types to control and coordinate lineage specification, cell proliferation, differentiation, migration, morphogenesis, and injury repair . Nearly all publications explore the transcriptomics, proteomics, or epigenetics of lung development. Three articles are reviews of existing published work, and four articles focus solely on bioinformatics and systems analysis work that has been accomplished by the LungMAP consortium. Disease conditions are included in 22 of the publications, which are focused on bronchopulmonary dysplasia and other complications of prematurity and idiopathic pulmonary fibrosis. Published work results include 14 that are specific to mouse experiments, 16 involving human studies, and eight that work with both mouse and human data. Although some human data is now offered on the LungMAP website, the abundance of human data that support the many published papers will be available for public sharing in early Fall of 2017.To date, LungMAP funds and efforts have contributed to the generation of data and tools used in 42 published manuscripts as well as numerous abstracts and posters at scientific conferences. Six of the journal articles reflect collaborative research by two or more LungMAP research centers Workshops on Cell Plasticity in Lung Injury and Repair in April 2010 and on Molecular Determinants of Lung Development in September 2011. The NHLBI Board of External Experts organized the LungMAP initiative to build a molecular atlas of late-stage lung development to serve as a platform for discovery research to better understand critical events, including alveologenesis. This atlas would integrate multiscale information of gene expression, 3D cellular structures, and key morphogenesis events to provide detailed molecular anatomy of the developing mouse and human lungs. 1) a mouse lung atlas from embryonic day 16.5 to postnatal day 28 that integrates gene expression, imaging, and anatomic analysis, 2) a human lung atlas to validate the molecular profile in lung cell types in normal human lungs from gestational week 23 to postnatal year 10, and 3) an integrated, publicly accessible, and expandable database to accommodate existing and newly generated data.The goals of the LungMAP initiative are to build The LungMAP consortium consists of a set of research centers, many of which represent multiple institutions with distinguishing expertise, engaged in collaborative activities to better understand the molecular processes that guide lung development, specifically during the stages of alveolar formation and development. This exploration is multipronged, including molecular , visual , and computational approaches. Research Centers are supported by a Human Lung Tissue Core and a Data Coordinating Center. The centers also participate in the sharing of resources such as mouse tissue and cells from a single well-controlled and characterized mouse colony operated by one of the LungMAP research centers. The resources currently supported by the LungMAP consortium that are available to the public are highlighted in The Cincinnati Children\u2019s Hospital Medical Center (CCHMC) Research Center (RC) is responsible for single cell transcriptomics and confocal imaging and, as a member of the Data Coordinating Center (DCC), is developing computer programs for specific analyses. The RC generates and analyzes detailed NextGen gene expression data for human and mouse lung, including from single dissociated cells. Although many of the genes and networks regulating lung formation are shared among vertebrate species, the physiology, structure, extracellular matrix, region-specific cell types, and gene expression patterns vary between murine and human lung , 25. To CCHMC is also producing a library of lung images, including immunofluorescent confocal as well as hematoxylin- and eosin-stained sections from mouse and human samples. Acting as the Mouse Hub for LungMAP, CCHMC also provides mouse tissues to all research centers to reduce variability in murine-based research due to differences in substrain or environmental conditions.The Pacific Northwest National Laboratory, Baylor College of Medicine, University of Washington, and Texas Advanced Computing Center together form a Research Center that applies high-throughput approaches to resolve the molecular changes of lung development specific to cell types and spatial location. This RC measures gene, protein, lipid, and metabolite levels in both mouse and human lung tissues. The multiomics characterization effort within this RC spans multiple scales from whole tissue to region-specific to cell-type specific analyses. State-of-the-art mass spectrometry (MS)-based proteomic, lipidomic, and metabolomic measurements are made on whole lung tissue samples, sorted cell populations , and laser-capture microdissected (LCM) alveolar septae , 4. SpatThe University of Alabama at Birmingham (UAB), Yale University, University of California-San Diego (UCSD), and Carnegie Mellon University (CMU) comprise a single research center focused on the systems biology of alveolar development. This center is generating a compendium of the dynamic and regional changes in DNA methylation and chromatin accessibility, microRNA, mRNA, and proteins that occur during alveolar formation to generate a dynamic temporal regulatory model of normal alveologenesis in both the mouse and human. Despite significant progress in understanding normal alveolar formation, a comprehensive understanding of the dynamic regulatory networks of this process is lacking. For example, although we are aware that the regulation of alveolar growth depends upon integration of numerous signals from multiple pathways , 20, 24,The Saban Research Institute at Children\u2019s Hospital of Los Angeles is the LungMAP 3D-imaging Research Center (RC), which is investigating alveolar development in unique ways that are leading to new insights . This RChttp://www.iiam.org) and the National Disease Research Interchange , organizations that link donors to the scientific community. A special effort, the Neonatal Organ Donation Program, was established by IIAM to provide donations to LungMAP and other important research efforts. IIAM and NDRI receive referrals to families wishing to donate for research when transplantation is not an option.The University of Rochester Medical Center, in collaboration with Seattle Children\u2019s Research Institute, operates the Human Tissue Core for LungMAP. The Human Tissue Core (HTC) identifies and manages tissue sources to meet the overall goal of LungMAP to build an open-access comprehensive molecular atlas of the late-stage developing lung. The HTC procures, processes, stores, and distributes normal late-gestation, neonatal, and early childhood human lung tissue and dissociated cells for the Research Centers, networking with the United Network for Organ Sharing and their partners. LungMAP studies the biology of normal lung development, which makes tissue health a critical concern. The HTC collects transplantation-quality tissues through the International Institute for the Advancement of Medicine that can be reconstructed to provide 3D images and then processed for histological analysis or dissociated and sorted to provide a range of enriched cell populations, including subsets of epithelial, endothelial, mesenchymal, lymphatic, immune, and stem cells. A working group of highly qualified pathologists work with the HTC to establish biorepository and assessment tools and review specimens to assure quality and consistent selection of the tissues chosen for LungMAP.https://brindl.urmc.rochester.edu) for tracking and identification of the derived products of the collection, including glass slides, paraffin and frozen blocks, cell fractions, RNA, digital CT and whole slide images, and associated donor data. The HTC LIMS system integrates with the LungMAP database for transfer of pertinent donor and tissue information. BRINDL provides complete specimen repository functions for tracking and managing requests to handle the traffic between the RCs and the HTC repository.The HTC maintains a laboratory information management system (LIMS) called BRINDL for LungMAP, with added bioinformatics expertise from CCHMC. The DCC developed and maintains the LungMAP website and database called Bioinformatics REsource ATlas for the Healthy lung (BREATH). The DCC processes data submissions from the RCs, integrates data into BREATH, works with centers on data display, and coordinates with the HTC. The DCC also serves as the administrative infrastructure to facilitate collaboration across the consortium, creating consortium-wide priorities and policies, communication plans, and coordinating working groups and steering committee meetings. The DCC is also responsible for, and with LungMAP working groups has already developed, greatly expanded mouse- and human-specific lung ontologies for data annotation and integration and for tools to enhance sharing of LungMAP data with the scientific community and the public.With the exponential growth of omics data, a major challenge of LungMAP is the determination and development of an effective way to integrate and to offer to the community the multiscale information that links anatomic structure and cell types with epigenetic, RNA, protein, lipid, metabolic, signaling, and molecular and cellular interaction data.Of fundamental importance to meet this challenge is the establishment of a database that can receive the complexity of data types generated by LungMAP and allow these data to be linked in myriad ways needed to investigate relationships. The DCC chose to build a triple-store database as the backbone of BREATH based on open-source OpenLink Virtuoso . Triple-www.lungmap.net) (A primary output to date for the DCC is the establishment and development of the LungMAP website (map.net) . The webmap.net) , Lung Gehttps://www.lungmap.net/resources/ontologies/), users can access the ontology through a special browser or download PDFs or Microsoft Word files containing the ontologies. These files include Human Alveolar Stage, Mouse E16.5 (early canicular stage), Mouse E18.5 [early saccular stage, Mouse P0\u2013P03 , Cell Ontology for Mouse Lung Maturation, and Cell Ontology for Human Lung Maturation. These documents are flat file views providing the LungMAP identification numbers for each term within the current deployed ontology .The LungMAP high-resolution anatomic ontologies describe the anatomic structures, tissues, and cells of the developing human and mouse lower respiratory tracts, including the trachea, bronchi, bronchioles, and alveolar parenchyma of the lung, as well as the pulmonary vascular, autonomic, and immune systems of the lung. The LungMAP ontology incorporates well-characterized structural and functional anatomic components, distinct histological tissue compartments, and generic and specific cell types. The ontology provides a coherent set of terms incorporated into an interactive, searchable, web-based atlas for the integration of anatomic, cellular, molecular, and imaging data. This ontology is organized along the proximal-distal axis of the lower respiratory tract, encompassing prenatal and postnatal stages of human and mouse lung development. On the LungMAP website (1) a web-based mechanism for image annotation, 2) a web-based mechanism to create \u201ctheme\u201d pages for display on LungMAP, and 3) a viewer to present image data in 3D. These efforts are in prototype development as of this writing, with products available to the public through the LungMAP website in 2017.Consortium efforts, led by the DCC, have focused on several specific components whose importance emerged as the project progressed. These include development of Imaging data provide the foundation upon which to build an atlas of the developing lung. The LungMAP consortium has created a library of lung images using multiple imaging technologies that span from anatomic to molecular. To enrich understanding of these visual data, it was necessary to have a mechanism for identifying specific areas of an image to highlight the structures and labeling of the tissue, including proteins, cells, and larger anatomic features such as bronchioles and vessels that develop at certain time points. The LungMAP Image Annotation Tool allows a user to overlay images with feature symbols , associated terms for structures, and notes. Annotators log into a secure site to access images in an environment that allows outlining of areas or placement of symbols onto the two-dimensional (2D) image with linkage to a term in the ontology. The tool is engaged by selecting any single image thumbnail from within an image experiment. Annotators manually place the symbol on the 2D image and add the additional information. The association of a symbol with the image (with descriptive information) is referred to as image annotation, as shown in A second approach to image annotation that LungMAP is developing uses machine-based algorithms that automatically identify specific components of the image. The development team has assembled a database of characteristics for 4,557 confocal immunofluorescent image files. These metadata are being used to group collections of images . They have successfully identified specific features of certain lung images by combining information from the metadata and image analysis routines from the OpenCV package was released for testing in late 2016, allowing researchers to build simple web pages within the LungMAP website. Ease of use was key to the design, with flexibility to incorporate text, image, and media to support presentation. Although in an early stage of design, plans for the next iteration include development of methods to incorporate LungMAP data directly into a constructed page for compilation and comparison of the many types of image and omics data. StoryBuilder will support consortium research as observations unfold and will provide a flexible forum for sharing as these conclusions evolve or change. StoryBuilder will allow researchers to support their observations with images, media links, and references to published literature (with proper acknowledgments/citations). Once made public on the LungMAP website, website viewers will have the opportunity to ask questions and post comments so that narratives are not static but can evolve as new research uncovers new discoveries.LungMAP offers a tool to visualize microscopy image data in a very unique way. An integrated input-output visualization tool called 3D MMVR (\u03bcCT-Microscopy Volume Renderer) does direct volume rendering using ray casting, a true volumetric representation of the data. Ray casting preserves detail and allows for the quick adjustment of opacities through controls in the user interface. Through the 3D MMMVR, a user can look inside lung structures to see internal airway passages, including cells contained within them. The 3D viewer opens a data set for the user to explore. Currently, the data are presented for adult mouse lung. All data are in a DICOM file structure. LungMAP will expand the use of this tool this year (2017) with more data and eventually invite outside researchers to use the tool with their own data to conduct analyses.LungMAP will continue to develop its lung data resource, adding additional data for both mouse and human samples. Expanded protein, lipid, and metabolomics data and single-cell expression and methylation data for additional key developmental time points are now being prepared for viewing and download. Models based on regulatory pathway analysis of lung development will be integrated into the website during this year (2017). Molecular and imaging data from lung researchers outside the LungMAP consortium and with databases such as Mouse Genome Informatics will be considered for downloads and integration with the BREATH database in 2018.In the coming year, we plan to increase engagement of the community through direct outreach. We will expand our beta tester group to give feedback on web functionality, offer seminar sessions to explore the data with LungMAP researchers, and further develop LungMAP for educational purposes through efforts such as enhancing our 3D renderer for wider public engagement. Bringing students and tech developers into the LungMAP community will ensure that LungMAP is relevant to a broad and diverse base of users for the present and years to come.The work was supported by the National Heart, Lung, and Blood Institute Grants U01-HL-122638 , U01-HL-122642 (to S. S. Potter and J. A. Whitsett), U01-HL-122703 , U01HL122700 , U01-HL-122626 , and U01-HL-122681 (to D. Warburton).No conflicts of interest, financial or otherwise, are declared by the authors.M.E.A.-P. and J.P.C. prepared figures; M.E.A.-P., R.F.C., and N.A. drafted manuscript; M.E.A.-P., R.F.C., C.A., J.P.C., J.S.H., G.S.P., J.A.W., and N.A. edited and revised manuscript; M.E.A.-P., R.F.C., C.A., J.P.C., R.A.C., G.H.D., J.S.H., N.K., T.J.M., S.S.P., G.S.P., D.W., J.A.W., S.M.P., N.A., and t.L.C. approved final version of manuscript."} +{"text": "Scientific Reports 10.1038/s41598-017-11969-5, published online 18 September 2017Correction to: The original version of this article contained an error in the spelling of the author Susan C Morpeth, which was incorrectly given as Susan Morpeth.Additionally, in this Article, an affiliation for Susan C Morpeth was omitted. The correct affiliations for Susan C Morpeth are listed below:KEMRI-Wellcome Trust Research Programme, Kilifi, KenyaLondon School of Hygiene & Tropical Medicine, London, UKThese errors have now been corrected in the PDF and HTML versions of the paper, and in the accompanying Supplementary Information."} +{"text": "In constraint-based metabolic modelling, physical and biochemical constraints define a polyhedral convex set of feasible flux vectors. Uniform sampling of this set provides an unbiased characterization of the metabolic capabilities of a biochemical network. However, reliable uniform sampling of genome-scale biochemical networks is challenging due to their high dimensionality and inherent anisotropy. Here, we present an implementation of a new sampling algorithm, coordinate hit-and-run with rounding (CHRR). This algorithm is based on the provably efficient hit-and-run random walk and crucially uses a preprocessing step to round the anisotropic flux set. CHRR provably converges to a uniform stationary sampling distribution. We apply it to metabolic networks of increasing dimensionality. We show that it converges several times faster than a popular artificial centering hit-and-run algorithm, enabling reliable and tractable sampling of genome-scale biochemical networks.https://github.com/opencobra/cobratoolbox.Bioinformatics online. The linear equalities constrain the system to a steady state where fluxes into and out of every node are balanced. Nonequilibrium steady-states are enabled by including metabolite sources and sinks, collectively known as exchange reactions, at the boundary of the system with the environment. The inequalities arise from physicochemical constraints such as thermodynamics, as well as environmental constraints such as nutrient availability. Fluxes can be further constrained to the optimal value A constraint-based model of a metabolic network, with Uniform sampling of constraint-based models . ACHR isS, l, u and c from CHRR consists of rounding followed by sampling see . The rouWhen sampling the feasible set of a constraint-based model, it is important to run the sampling algorithm until the sampling distribution converges to a stationary distribution of fluxes over \u03a9. Otherwise, the sampling distribution is likely to be misrepresentative, leading to incorrect conclusions about the model see . It is gWe compared the convergence time of CHRR to the COBRA toolbox implementation of ACHR . We founCoordinate hit-and-run with rounding makes uniform sampling of genome-scale metabolic networks tractable and reliable. The compatibility of our implementation with the COBRA toolbox should facilitate widespread utilization by the constraint-based metabolic modelling community.HSH and IT were supported by the Luxembourg National Research Fund (FNR) through the National Centre of Excellence in Research (NCER) on Parkinson\u2019s disease. BC and SV were supported in part by NSF awards CCF-1217793 and EAGER-1415498. RMTF was funded by the Interagency Modeling and Analysis Group, Multi-scale Modeling Consortium U01 awards from the National Institute of General Medical Sciences, award GM102098, and U.S. Department of Energy, Office of Science, Biological and Environmental Research Program, award DE-SC0010429.Conflict of Interest: none declared.Supplementary DataClick here for additional data file."} +{"text": "Reference 29 is cited in error in the published article. Reference 29 should be removed from the article. There is also an error in Reference 11 in the published article. The correct reference is: Leyvraz M, Wirth JP, Woodruff BA, Sankar R, Sodani PR, Sharma ND, et al. High Coverage and Utilization of Fortified Take-Home Rations among Children 6\u201335 Months of Age provided through the Integrated Child Development Services Program: Findings from a Cross-Sectional Survey in Telengana, India. PLoS ONE. Public Library of Science; 2016."} +{"text": "Self is difficult to define because of its multiple, constitutive streams of functional existence. A more comprehensive and expanded definition of self is proposed. The standard bio-psycho-social model of psyche is expanded to biophysical-mental-social and existential self. The total human experience is better understood and explained by adding the existential component. Existential refers to lived human experience, which is firmly rooted in reality. Existential living is the capacity to live fully in the present, and respond freely and flexibly to new experience without fear. Four common fears of isolation, insecurity, insignificance, and death can be overcome by developing a lifestyle of whole-hearted engagement in the present reality, creative problem solving, self-actualization, and altruism. Such integrative living creates a sense of presence with self-awareness, understanding, and existential well-being. Well-being is defined as a life of happiness, contentment, low distress, and good health with positive outlook. Self is a complex, integrative process of living organisms. It organizes, coordinates, and integrates energy-information within and around itself, spontaneously, unconsciously, and consciously. Self-process is understood in terms of synergetics, coordination dynamics, and energy-information\u2013directed self-organization. It is dynamic, composite, ever renewing, and enduring. It can be convergent or divergent, and can function as the source or target of its own behavior-mentation. The experience of self is continuously generated by spontaneous activation of neural networks in the cerebral neocortex by the brainstem-diencephalic arousal system. The multiple constitutive behavioral-mental streams develop concurrently into a unique experience of self, specific for a person at his/her developmental stage. The chronological neuro-behavioral-mental development of self is described in detail from embryonic stage to old age. Self can be behaviorally-mentally oriented and realized in three complimentary modes of being: egocentric, allocentric, and ecosystemic or existential. The existential mode is both immanent and transcendent, and can be self-actualized, resulting in a healthy, creative, conflict-free, and meaningful life."} +{"text": "The South-East Asia Region Kala-azar Elimination Programme (KAEP) is expected to enter the consolidation phase in 2017, which focuses on case detection, vector control, and identifying potential sources of infection. Post-kala-azar dermal leishmaniasis (PKDL) is thought to play a role in the recurrence of visceral leishmaniasis (VL)/kala-azar outbreaks, and control of PKDL is among the priorities of the KAEP.We reviewed the literature with regard to PKDL in Asia and interpreted the findings in relation to current intervention methods in the KAEP in order to make recommendations. There is a considerable knowledge gap regarding the pathophysiology of VL and PKDL, especially the underlying immune responses. Risk factors (of which previous VL treatments may be most important) are poorly understood and need to be better defined. The role of PKDL patients in transmission is largely unknown, and there is insufficient information about the importance of duration, distribution and severity of the rash, time of onset, and self-healing. Current intervention methods focus on active case detection and treatment of all PKDL cases with miltefosine while there is increasing drug resistance. The prevention of PKDL by improved VL treatment currently receives insufficient attention.PKDL is a heterogeneous and dynamic condition, and patients differ with regard to time of onset after VL, chronicity, and distribution and appearance of the rash, as well as immune responses , all of which may vary over time. It is essential to fully describe the pathophysiology in order to make informed decisions on the most cost-effective approach. Emphasis should be on early detection of those who contribute to transmission and those who are in need of treatment, for whom short-course, effective, and safe drug regimens should be available. The prevention of PKDL should be emphasised by innovative and improved treatment for VL, which may include immunomodulation. Kala-azar/visceral leishmaniasis (VL) is endemic in the Indian subcontinent (ISC), affecting the Gangetic plains of India, Bangladesh, and Nepal.Phlebotomus argentipes being the only vector and susceptibility to insecticide; the availability of an accurate rapid diagnostic test (RDT) detecting antibodies against the 39-aminoacid-recombinant kinesin antigen (anti rK39); the use of an oral drug, miltefosine; and strong political commitment from the 3 countries . A . A 16]. Short, ambulant treatment courses are needed, preferably with (an) oral drug(s). The current treatment with miltefosine is of long duration, and there are reports of increasing failure . For anyFor the design of a cost-effective control strategy, the contribution of each category of PKDL patients should be studied: acute versus chronic, macular versus papular versus nodular, and limited versus extensive disease. In each clinical presentation, infectivity of the blood should also be studied. This can only be done by xenodiagnosis studies that quantify the potential for infectivity. For this purpose, insectaria have been established at Muzaffarpur (India) and Mymensingh (Bangladesh).Clearly, this information is essential for the design and application of mathematical models in which the potential contribution of PKDL to transmission should be balanced and quantified against that of other clinical entities in the spectrum of VL: VL cases, ex-VL cases, HIV\u2013VL coinfection, and asymptomatic individuals. Also, here, understanding the dynamics is important because the contribution of each may vary in the course of the condition as well as over time in the local epidemiological context ,77. On tOther epidemiological information such as seasonality of sand flies and their behaviour, human migration, and the potential increase of HIV coinfection are among other factors to be explored. Dye and Wolpert recognised the influence of factors such as earthquakes, influenza, and malaria on the statistics of fever (VL or other) deaths, thus emphasising the dynamics over time .In conclusion, the current optimal results obtained so far in the KAEP are fragile and must be consolidated before commitment is lost. History has shown that PKDL is a crucially important factor in the control of VL in the ISC. For the short term, a strategy for active case finding; the validation and implementation of diagnostic tools such as LAMP or qPCR; optimal treatment for VL aiming at the lowest possible PKDL rates; and the identification of short, safe, and effective treatment for PKDL are essential for the VL elimination efforts in the ISC to succeed and be sustained. For the long term, immunological studies and infectivity studies may provide insight in more targeted and more efficient approaches.The immune responses at various manifestations of PKDL need to be described .The contribution of various manifestations of PKDL in transmission needs to be studied in xenodiagnosis studies and compared to the contribution of asymptomatics, VL patients, HIV-coinfected VL patients, and ex-VL patients, including parasite load by qPCR.Prevention of PKDL by optimal VL treatment is crucial.Optimal treatment for PKDL should be based on a safe oral drug with good skin penetration and immunomodulatory properties; it may include an immunomodulator.A point-of-care biomarker for cure needs to be identified to define the optimal duration of treatment for PKDL.Banjara MR, Kroeger A, Huda MM, Kumar V, Gurung CK, et al. (2015) Feasibility of a combined camp approach for vector control together with active case detection of visceral leishmaniasis, post kala-azar dermal leishmaniasis, tuberculosis, leprosy and malaria in Bangladesh, India and Nepal: an exploratory study. Trans R Soc Trop Med Hyg 109: 408\u2013415.Hirve S, Boelaert M, Matlashewski G, Mondal D, Arana B, et al. (2016) Transmission Dynamics of Visceral Leishmaniasis in the Indian Subcontinent\u2014A Systematic Literature Review. PLoS Negl Trop Dis 10: e0004896.WHO. (2011) Regional Strategic Framework for Elimination of kala-azar from the South-east Asia Region (2011\u20132015). SEA-CD-239.http://apps.who.int/iris/bitstream/10665/78608/1/9789241505215_eng.pdf .WHO. Post-kala-azar dermal leishmaniasis: a manual for case management and control. Zijlstra EE (2016) The immunology of post-kala-azar dermal leishmaniasis (PKDL). Parasit Vectors 9: 464S1 Table(DOCX)Click here for additional data file.S1 Fig(DOCX)Click here for additional data file."} +{"text": "The late life disability instrument (LLDI) was developed to assess limitations in instrumental and management roles using a small and restricted sample. In this paper we examine the measurement properties of the LLDI using data from the Lifestyle Interventions and Independence for Elders Pilot (LIFE-P) study.LIFE-P participants, aged 70-89 years, were at elevated risk of disability. The 424 participants were enrolled at the Cooper Institute, Stanford University, University of Pittsburgh, and Wake Forest University. Physical activity and successful aging health education interventions were compared after 12-months of follow-up. Using factor analysis, we determined whether the LLDI's factor structure was comparable with that reported previously. We further examined how each item related to measured disability using item response theory (IRT).The factor structure for the limitation domain within the LLDI in the LIFE-P study did not corroborate previous findings. However, the factor structure using the abbreviated version was supported. Social and personal role factors were identified. IRT analysis revealed that each item in the social role factor provided a similar level of information, whereas the items in the personal role factor tended to provide different levels of information.Within the context of community-based clinical intervention research in aged populations, an abbreviated version of the LLDI performed better than the full 16-item version. In addition, the personal subscale would benefit from additional research using IRT.http://www.ClinicalTrials.gov (registration # NCT00116194).The protocol of LIFE-P is consistent with the principles of the Declaration of Helsinki and is registered at Disability is a major focus for intervention research in aging due to the social, personal, and economic costs associated with the loss of independence . The magThe Lifestyle Interventions and Independence for Elders Pilot (LIFE-P) study was a single blind four-center randomized controlled trial of a 12-month physical activity (PA) intervention compared to a successful aging (SA) intervention in sedentary older adults. The LLDI was used to measure change in disability within randomized groups of LIFE-P. Because the original LLDI was developed on a small, restricted sample, prior to measuring change in the LLDI within LIFE-P, we undertook an investigation to re-examine the measurement properties of the instrument. The longitudinal design of LIFE-P enabled us to examine the stability of the factor structure of the LLDI as disability responsive to change with time and to evaluate the quality of individual items.We initially use confirmatory factor analysis to investigate whether the factor structure for the limitation domain of the LLDI, as applied to baseline and follow-up data obtained from LIFE-P participants, was compatible with the originally publication. Furthermore, because McAuley and colleagues publisheIn LIFE-P, at baseline, 6- and 12-months, comprehensive standard assessments were conducted by trained research staff blinded to intervention assignment -7. The sdoing a particular task?\" with response options of \"not at all,\" \"a little,\" \"somewhat,\" \"a lot,\" and \"completely.\" Jette et al. [The Late Life Disability questionnaire includes items for a wide variety of life tasks, such as personal maintenance; mobility and travel; exchange of information; social, community, and civic activities; home life; paid or volunteer work; and involvement in economic activities . It was e et al. demonstre et al. identifiWe obtained data on participant's age, gender, race/ethnicity, education, marital status, and living arrangements using a structured personal interview. Prevalence of clinical conditions, including heart condition, chronic pulmonary condition, anxiety/depression, stroke, diabetes, high blood pressure, hip fracture, liver disease, and cancer, was determined using self-reported physician-diagnosed disease information . The meaParticipant Characteristics in the LLDI developmental sample and the LIFE-P at Baseline were compared. Percentage was presented for categorical variables and mean was presented for continuous variables.We compared our LIFE-P factor solutions with those from Jette et al and McAuSubsequently, we applied Confirmatory Factor Analysis (CFA) at baseline, 6-months, and 12-months to check whether the factor structure for the limitation domain from the LLDI was compatible with the original publications ,4. MaximAs an item-level exploration, we applied IRT analysis within each factor for the 12-month data. The month 12 visit was selected because at that visit participants exhibited a wider amount of variation in level of disability and we reasoned that data from this visit might more closely resemble the samples used in previous publications. For easier interpretation purpose, we divided the scale for each limitation item into the following two groups: the \"less limitation\" classification included responses of \"not at all,\" \"a little,\" and \"somewhat\", whereas the \"a lot of limitation\" classification included responses of \"a lot,\" and \"completely\". Item parameters were generated including difficulty (location) and discrimination (slope or correlation) [Table The study design, recruitment, and participant characteristics of McAuley et al. have beeThere were not many missing LLDI items in the LIFE-P study; the rates of missing items were below or equal to 1% for all items except one (\"work at a volunteer job\" at baseline) was 2%. Results from EFA are presented in Table Since the result of our factor analysis was not comparable to that reported by Jette et al., we further applied EFA to the eight items (the abbreviated version) reported by McAuley et al. . AdoptinResults from the CFA for the limitation domain of the LLDI from LIFE-P are provided in Table IRT was subsequently used to empirically assess the relation between the factor and each of the four items (abbreviated version) that loaded highly on the specific factor at month 12 in the LIFE-P participants. Results from this analysis are presented in Figures The factor structure for the limitation domain using the 16 items within the LLDI in LIFE-P study did not corroborate the findings reported by Jette et al . The twoThere are several possible reasons why we were unable to confirm the originally published factor structure of the LLDI. First, because the sample size from the LLD developmental sample was small, those results may be unstable. Ideally, the LLDI should be evaluated in large, population-based samples. Second, LIFE-P was a community-based clinical trial and the study participants may not be representative of the LLDI developmental sample. For example, from Table The item-level analysis indicates that the level of information for social roles provided by each of the four items was consistent, showing that the stated activities are of equal importance in capturing late life activities. However, items on the second factor - personal role - tend to provide different levels of information. For example, most participants seem to be able to take care of essential household business, as reflected in the low difficulty item parameter and low information of the household business item. However, participants may not have the capacity or willingness to perform non-essential local errands.So what is the take home message and where should research with the LLDI go from here? First, we see no advantage of using the long form over the short form and would suggest that investigators use the brief 8-item LLDI in future research. Second, application of item-response theory to the LLDI short form offered support for the content of the social subscale, but it was mixed for items making up the personal subscale. Future research is needed with the personal subscale in populations that have greater difficulty with basic activities of daily living (ADLs). In particular, even though the physical functioning of LIFE-P participants was compromised somewhat, these individuals did live independently in the community. The personal subscale may be more appropriate for studies conducted within senior living communities in which older adults often have difficulty with one or more basic ADL. This also raises the more general issue of using the LLDI in both large epidemiological studies and smaller controlled trials. Unless the population of interest involves older adults that either have or are likely to experience deficits in functioning that compromise very basic social and personal activities, the LLDI should not be used. Third, LIFE-P collected the LLDI at three different time points: application of factor analysis to each time point may not be the most efficient way to evaluate the properties of the questionnaire. Accordingly, it is crucial for methodologists to develop methods that can incorporate the factor data at different time points while considering the possible different factor structure at each time point.In summary, we contrasted LLDI results from LIFE-P and two other studies ,4. The aNo other potential competing interest relevant to this article was reported.FCH, WJR, EHI, and AMJ: study concept and design, analysis and interpretation of data, preparation of manuscript. JAK and RF: study concept and design, preparation of manuscript. SAS: acquisition of data. SNB and MEM: acquisition of data, study concept and design, analysis and interpretation of data, preparation of manuscript. All authors read and approved the final manuscript.The Lifestyle Interventions and Independence for Elders Pilot (LIFE-P) Study was funded by a grant from the National Institutes of Health/National Institute on Aging (U01 AG22376) and supported in part by the Intramural Research Program, National Institute on Aging, NIH. The Wake Forest University Field Center was partially supported by the Claude D. Older American Independence Pepper Center (1P30AG21332). Dr. Fielding's contribution was partially supported by the U.S. Department of Agriculture, under agreement No. 58-1950-4-401. The Pittsburgh Field Center was partially supported by the Pittsburgh Claude D. Pepper Center P30 AG024827.The Lifestyle Interventions and Independence for Elders Study Group: Cooper Institute, Dallas, TX: Steve Blair, Timothy Church, Jamile A. Ashmore, Judy Dubreuil, Alexander N. Jordan, Gina Jurca, Ruben Q. Rodarte, Jason M. Wallace; National Institute on Aging: Jack M. Guralnik, Evan C. Hadley, Sergei Romashkan; Stanford University, Palo Alto, CA: Abby C. King, William L. Haskell, Leslie A. Pruitt, Kari Abbott-Pilolla, Karen Bolen, Stephen Fortmann, Ami Laws, Carolyn Prosak, Kristin Wallace; Tufts University, Boston, MA: Roger Fielding, Miriam Nelson; University of California, Los Angeles, Los Angeles: Robert M. Kaplan; University of California, San Diego: Eric J. Groessl; University of Florida, Gainesville: Marco Pahor, Connie Caudle, Lauren Crump, Tonya Kelley; University of Pittsburgh, PA: Anne B. Newman, Bret H. Goodpaster, Stephanie Studenski, Erin K. Aiken, Steve Anthony, Nancy W. Glynn, Judith Kadosh, Piera Kost, Mark Newman, Christopher A. Taylor, Pam Vincent; Wake Forest University, Winston-Salem, NC, Field Center: Stephen B. Kritchevsky, Peter Brubaker, Jamehl Demons, Curt Furberg, Jeffrey A. Katula, Anthony Marsh, Barbara J. Nicklas, Kimberly Kennedy; Shruti Nagaria, Rose Fries, Katie Wickley-Krupel; Data Management and Quality Control Center: Michael E. Miller, Mark A. Espeland, Fang-Chi Hsu, Walter J. Rejeski, Don P. Babcock, Jr., Lorraine Costanza, Lea N. Harvin, Lisa Kaltenbach, Wesley A. Roberson, Julia Rushing, Michael Walkup; Yale University, New Haven, CT: Thomas M. Gill."} +{"text": "Correction to: British Journal of Cancer (2009) 100, 1889\u20131895; doi:10.1038/sj.bjc.6605093Owing to an error during typesetting, the name of the third author, Richard Morgan, was omitted from this paper when first published in the June 2009 issue of BJC. The publishers would like to apologise for this mistake."} +{"text": "We describe a case of a 25 year old female from Lithuania who presented with a productive cough. Chest radiograph demonstrated an infiltrate in the left upper lobe and a cavitating lesion in the right middle lobe. Sensitivity testing of her sputum led to a diagnosis of extensively drug-resistant tuberculosis (XDR-TB). This is the first case in Ireland and highlights the need for physicians to be aware of the possibility of XDR-TB. Moreover it underlines the need for improvement in service provision in terms of a TB reference laboratory and TB clinics. Diagnostic investigations confirmed acid fast bacillus in the sputum and she was commenced on anti-tuberculous treatment. Nine days later TB culture demonstrated resistance to all first line agents. She was commenced on oral moxifloxacin, prothionimide, PAS, cycloserine and intramuscular capreomycin. She was discharged home on Directly Observed Therapy (DOT) in close conjunction with our Public Health colleagues.A 25-year-old female, non-smoker, originally from Lithuania who had been living in Ireland for 2 years presented with a two month history of cough productive of brown sputum. She had no weight loss, night sweats, or dyspnoea. The full blood count was normal, HIV test negative and chest radiograph demonstrated an area of infiltration in the left upper lobe and a cavitating lesion of the right middle lobe , was reported and demonstrated sensitivity to capreomycin, clofazimine, prothionimide and cycloserine. Resistance was noted to amikacin, streptomycin, isoniazid, rifampicin, ethambutol, pyrazinamide, ciprofloxin, clarithromycin, rifabutin and PAS. Second line susceptibility were performed in radiometric Bactec\u2122 in 2 reference laboratories. In view of the resistance, treatment was continued on the 4 drug regimen: moxifloaxin, cycloserine, prothionimide and capreomycin. She became smear negative after 2 months of therapy. After eight months she developed tinnitus and the capreomycin was stopped, audiogram was normal. She received DOT therapy for a total of 20 months. Chest radiograph following 20 months of therapy demonstrated no evidence of active disease or evidence of old fibrocalcific disease.Five weeks following initial diagnosis, the extensively drug-resistant tuberculosis (XDR-TB) profile (Table Mycobacterium tuberculosis that were resistant not only to isoniazid and rifampicin (i.e. MDR-TB) but also to at least three of the six classes of second-line anti-TB drugs . XDR-TB was redefined at a meeting of the WHO XDR-TB Task Force, in October 2006, in Geneva, such that it is now defined as: resistance to at least rifampicin and isoniazid (which is the definition of MDR-TB), in addition to any fluoroquinolone, and to at least one of the three following injectable drugs used in anti-TB treatment: capreomycin, kanamycin and amikacin [XDR-TB was first described as an entity in March 2006, in a report jointly published by the US Centers for Disease Control and Prevention (CDC) and WHO . At this. In Asia, data are available from South Korea [Because of its recent description, there is a paucity of published data on the prevalence of XDR-TB. In the USA, 4% of the MDR-TB strains isolated between 1993 and 2004 were XDR-TB. In Europe, representative data are available from Latvia [th Korea , where 1th Korea where 12th Korea . Furtherth Korea .The largest European study on XDR-TB has recently been published and it highlights two important points firstly XDR-TB cases are frequently not identified in clinical practice, and of even greater concern that XDR-TB cases have a clinical outcome worse than MDR-TB. This indicates that the XDR diagnosis has important treatment and prognostic values . AttentiCorrect and complete treatment of MDR-TB and XDR-TB is vital both for the individual patient and the community. There are no published guidelines on the management of XDR-TB. Guidelines on the management of MDR-TB suggest use of 4-6 drugs ,12 althoThe emergence of MDR and XDR-TB in Ireland requires us to take several measures: These include offering TB disease and Latent TB infection (LTBI) screening, followed by treatment when appropriate, to all new entrants from high TB endemic countries. Additionally, we should improve our laboratory capacity for susceptibility testing of second line drugs. We can also improve our Public Health service by ensuring more rapid contact tracing and medication compliance, though easy access to DOT. There is now a responsibility on medical and scientific TB personnel to develop methods to rapidly diagnose smear-negative TB cases, to evaluate new markers of TB infection and disease, to develop new drugs for the treatment of TB and to develop international standards and guidelines for the management of drug resistant TB.This case highlights an emerging health problem in Europe and underlines the necessity for physicians to be aware of MDR and XDR-TB in patients and most particularly in patients presenting from areas of high prevalence."} +{"text": "To the Editor: Spread of drug-resistant tuberculosis (TB) and disastrous rates of HIV-TB co-infection pose serious threats to TB-control programs around the world , all isolates were found to belong to the M. tuberculosis complex. Testing of isolates for susceptibility to isoniazid, rifampicin, ethambutol, and streptomycin was performed by using the standard Mycobacteria Growth Indicator Tube manual system, as recommended by the manufacturer . The Wayne assay (M. tuberculosis complex (M. tuberculosis, i.e., resistant to both isoniazid and rifampicin.In 1997 the national TB programs of Myanmar introduced DOTS in the capital city, Yangon, which has approximately 5 million inhabitants. All new case-patients in national TB program clinics are routinely treated with isoniazid, rifampicin, ethambutol, and pyrazinamide without drug susceptibility testing of The World Health Organization/International Union Against Tuberculosis and Lung Diseases global survey in the year 2000 (M. tuberculosis isolates from Myanmar.The low number of MDR cases in our study could partly be explained by demographic features of the studied population, which is composed predominately of people residing in satellite townships of Yangon. These townships usually attract young people who immigrate to Yangon from village areas. These immigrants are less likely to have previous exposure to TB than the permanent population since the prevalence of TB infection is lower in rural than in urban areas ("} +{"text": "After publication of this work , we founAccordingly, some parts in \"Samples and DNA preparation\" of Methods, Authors' contributions and Acknowledgements have been revised as follows:Shinkaia crosnieri was used for the experiments. Sampling was conducted using a suction sampler with multiple canisters installed on the ROV Hyper-Dophin belonging to the Japan Agency for Marine-Earth Science and Technology (JAMSTEC) on Apr 24, 2005. [The followings are the same.]One specimen of J-SY designed and performed the experiments, analyzed the data, and wrote the paper. HN participated in DNA preparation. YF and ST collected the materials. W-JY directed the research and paper writing. All authors contributed to the manuscript and then read and approved the final version."} +{"text": "Some information was omitted from the funding statement. The correct statement should read: This work was funded by the Leukaemia Research Fund , Marie Curie Transfer of Knowledge (ToK) award (MOTET), and the NERC Fellowship to MRV. (NER/J/S/2002/00618). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "The aim of this study was to estimate the costs of treatment of children who present with the signs and symptoms of invasive bacterial diseases in Khanh Hoa province, Viet Nam. The study was an incidence-based cost-of-illness analysis from the health system perspective. The hospital costs included labour, materials and capital costs, both direct and indirect. Costs were determined for 980 children, with an average age of 12.67 months (standard deviation\u00b111.38), who were enrolled in a prospective surveillance at the Khanh Hoa General Hospital during 2005-2006. Of them, 57% were male. By disease-category, 80% were suspected of having pneumonia, 8% meningitis, 3% very severe disease consistent with pneumococcal sepsis, and 9% other diseases. Treatment costs for suspected pneumonia, meningitis, very severe disease, and other diseases were US$ 31, US$ 57, US$ 73, and US$ 24 respectively. Costs ranged from US$ 24 to US$ 164 across different case-categories. Both type of disease and age of patient had statistically significant effects on treatment costs. The results showed that treatment costs for bacterial diseases in children were considerable and might differ by as much as seven times among invasive pneumococcal diseases. Changes in costs were sensitive to both age of patient and case-category. These cost-of-illness data will be an important component in the overall evidence base to guide the development of vaccine policy in Viet Nam. Streptococcus pneumoniae causes half of all deaths due to pneumonia in children, and 70% of these deaths occur in Africa and Asia . Besidesand Asia . A largeand Asia ,5. Howevand Asia \u20138. To adWe conducted an incidence-based cost-of-illness analysis from the healthcare system perspective, a technique used for measuring the economic burden of patients from the onset to the end of illness ,10. We sFor our analysis, cost-of-illness was defined as the sum of direct medical, direct non-medical, and indirect costs . Direct To compute costs of each medical service, we used the ratio of costs to charges (RCCs) method ,16. In tDemographic information was described by frequency and proportion. We calculated means with standard deviations (SDs) for continuous variables and used multiple regression analysis to explore the factors associated with variations in treatment costs. Covariates considered in this modelling included age (months), duration of stay (days), and diagnosis. In addition, since prices of drugs may vary substantially among hospitals, we performed a sensitivity analysis to explore the effect of variation in prices of antibiotics across a range of prices of antibiotics available from the KHGH databases.Patients\u2019 demographics are presented in To convert the payment to economic cost, the cost-to-charge ratio method was applied. The KHGH had total costs of VND 172,482 million with revenue of VND 151,219 million or a hospital cost-to-charge ratio of 1.14. Of 48 antibiotics used for treating the study patients, ceftriaxone, cefotaxime, and cefuroxime were the most expensive drugs . These t2=0.172). Estimated costs regarding age and diagnosis were constructed. The costs of all disease-groups were inversely related to age\u2014as the age of patients increased, costs decreased , labour (28%), and capital (18%) were all similar to those of Thailand \u201329. For 2=0.172), other variables not included in our modelling procedures may explain variations in cost. In addition, the relationship of the independent variables may be confounded by age. For example, younger children may be more vulnerable to pneumococcal diseases than older children. In addition, drug-prescribing behaviours may affect both use of certain drugs and costs of drugs provided (In general, consumption of drugs was positively associated with both body-weight and age. Due to the relatively low explanatory power of the fitted cost model (adjusted Rprovided . We wereWe believe that data used in our analysis are both reliable and internally valid. The KHGH has a well-organized computerized database, from which all costing data were drawn. Our findings regarding age- and disease-specific treatment costs can be used together with age-specific incidence rates for such economic evaluations as cost-effectiveness, cost-utility, and cost-benefit analyses . Age-speTreatment cost of pneumococcal diseases was considerable in Viet Nam. Costs of suspected/probable and confirmed pneumococcal disease were statistically different as were costs of pneumococcal pneumonia and meningitis, which can differ by as much as seven times. Age of patient and type of disease affect costs of treating pneumococcal diseases in Viet Nam. These data will be important in the overall evidence to guide the development of vaccine policy in Viet Nam.The study was supported by the PneumoADIP and by the Governments of Kuwait, Sweden, and South Korea.The authors thank Dr. Le Thi Phuong, Mr. Nguyen Thanh Hien, Mr. Nguyen Vu Cuong, and Mr. Van Hoi for superb work in data collection and assistance for data management and Kathy Murray for editorial assistance."} +{"text": "The first and second authors of this article, John A. Mengshol and Lucy Golden-Mason, should be listed as having contributed equally to this work."} +{"text": "Journal of Translational Medicine (JTM) established \"The Excellence in Translational Medicine Award\" in 2006 [In a continuing endeavor to recognize outstanding contributions in the field of translational medicine, the Editorial Board of the in 2006 . With thPfizer Global Research and Development and Global Translational Medicine and $2,500 contributed anonymously). The recipient of \"The Bedside-to-Bench Award\" will receive $5,000, generously provided anonymously. The funds received from each Award are to be used to cover expenses for any meeting sponsored by a non-for-profit organization that is relevant to the goal of translational medicine and research.The recipient of \"The Excellence in Translational Medicine Award\" will receive a $4,000 prize, from investigators representative of nine countries of four continents, covering a wide range of disciplines published in JTM between 1 July 2007\u201330 June 2008 were evaluated. For this purpose, an Award Committee* comprised of ten members of the Editorial Board and one non-editorial board member selected and co-chaired by Richard J. Ablin and Pier Giorgio Natali was formed. The National Institutes of Health Scoring System of 1\u20135, with 1 = Outstanding and 5 = Poor were used with the papers being evaluated with regard to their:\u2022 Scientific merit\u2022 Originality\u2022 Clarity\u2022 Relevance to the purposes of translational medicine and research (and in \"The Bedside-to-Bench Award\" to direct study of human subjects)\u2022 Research design\u2022 MethodologyGiven the papers by Ying Jiang , Merck RWith the necessity in drug development to make appropriate and cost effective \"go\" vs. \"no go\" decisions based on safety, Jiang et al. demonstrCorticosteriods, prolonging ambulation, provides a limited, at best treatment option for Duchenne muscular dystrophy (DMD). Multiple treatment options under development include gene replacement therapy. In the report by Rodino-Klapac et al. , co-reciRegulatory T cells , fundamental in maintaining tolerance to self-antigens, can thwart T cell immunity to tumour-associated antigens and thereby, represent a major obstacle to immunotherapy. Thus, reducing their number or inhibiting their effector functions intuitively has the potential of increasing the efficacy of anti-tumour immunity. While increasing preclinical data support this hypothesis, appropriate proof of concept trials in man are yet to be demonstrated.de novo appearance of melanoma antigen-specific CD8+ T cells. By extensively relying on the analysis of patient samples, the study by Rasku et al. [The contribution of Mary Ann Rasku et al. , Graham u et al. , represeu et al. exemplif2nd \"Excellence in Translational Medicine\" and 1st \"Bedside-to-Bench\" Awards are now history. We are hopeful these Awards will serve to encourage other investigators devoted to improving the \"bench-to-bedside\" and \"bedside-to-bench\" concepts of translational medicine and respective initiatives. Competition is now open for the subsequent Awards in each of the two categories in which, we very much look forward to the opportunity of selecting next year's winning papers.With congratulations to Ying Jiang , Louise *\"Excellence in Translational Medicine and Bedside-to-Bench Awards Committee\": Richard J. Ablin (Co-Chairman); Jean-Pierre Armand; Howard L. Kaufman; Bruce Litman; Pier Giorgio Natali (Co-Chairman); Hideho Okada; Michael Perricone; Rja K. Puri; Noriyuki Sato; Patrick F. Terry and Craig Webb."} +{"text": "Chair of Scientific Committee:Dr. Athanasios I. Zavras, Harvard University \u2013 ISPTID PresidentDr. Matthew Falagas, Tufts Univ. & Dunant HospitalScientific Committee Members:Dr. Cynthia Boschi, World Health OrganizationDr. Carolyn Dresler, International Agency for Research on Cancer, WHOProf. Anthony Hedley, University of Hong Kong School of MedicineAsst. Prof. Taru Kinnunen Mustonen, Harvard University School of DentistryProf. Steve Sussman, Univ. of California San FranciscoProf. David Scott, Univ. of Louisville School of Dental MedicineDr. Alexandra Shields, Georgetown University Institute for Health PolicyOrganizing Committee Members:Prof. Konstantinos Alexandridis, Univ. of Athens School of Dental MedicineAsst. Prof. Vassillis Gorgoulis, Univ. of Athens School of MedicineDr. Argyris Michalopoulos, Intensive Care Unit, Dunant HospitalDr. Katerina Katsampe, Deputy Mayor of AthensProf. John Kyriopoulos, National School of Public HealthMr. Kyriakos Souliotis, Onasion Cardiothoracic Surgery HospitalMr. Dimitrios Zavras, National School of Public HealthDear DelegatesIt is a great honor to host the 4th annual meeting of the International Society for the Prevention of Tobacco Induced Diseases in Athens, Greece. As the birthplace of democracy, Athens seems the ideal place for debate. And the subject, quantifying tobacco's impact, seems most suitable for scientific exchange.Tobacco use is the single most important preventable risk factor for a number of debilitating diseases around the world. ISPTID global; membership is dedicated to the prevention of tobacco induced diseases. Some of the primary aims of the annual ISPTID conference are a) to disseminate important, state-of-the-science information about the biologic effects and the public health impact of tobacco use, b) to present updated data about the clinical signs and symptoms of tobacco induced disease and the various methods of smoking control and prevention, c) to create and facilitate research networks, and d) to educate healthcare professionals in tobacco control.Financial interests, trade issues and taxation have allowed tobacco's spread for decades. The international community has recently agreed on a number of measures to curb tobacco use by adopting the first ever public health treaty, the Framework Convention of Tobacco Control (FCTC). The first indications of FCTC success around the world will be presented at the conference.Apart from their strong international presentations, ISPTID conferences attempt to contribute to the promotion of tobacco control locally. Given the high prevalence of smoking in Greece, the Society will address the local media and will host a workshop for healthcare professionals on how to design and implement tobacco cessation programs in their prospective clinical settings.We hope you enjoy the meeting.For better health,Athanasios I. Zavras, DMD, MS, DrMscISPTID PresidentMatthew Falagas, MD, MPHChair of the Scientific CommitteeThe authors declare that they have no competing interests.Please see additional file 1Click here for file"} +{"text": "A historical prospective study was conducted at the Mercy Hospital of Pittsburgh, Pennsylvania (USA), to study the role of post-menopausal obesity in the recurrence and survival of breast cancer. Records from 301 post-menopausal women diagnosed with breast cancer from 1977 to 1985 were followed for at least 5 years from data supplied by the Tumor Registry and medical records. Data collected included age, height, weight, race, hormone receptor status, stage and size of tumour, number of positive nodes, site of distant metastasis, first course of treatment, and 5 year recurrence and survival. Forty-five per cent of patients were obese (n = 136), while 55% were non-obese (n = 165). Obesity was defined by the Quetelet index . The recurrence rates for the obese and non-obese groups were 40% and 39% respectively, and were not significantly different. Univariate and multivariate analyses showed that there was no significant association between obesity in post-menopausal women and likelihood of recurrence of or death from breast cancer."} +{"text": "Ecologic and economic factors, as well as changes in human behavior, have resulted in the emergence of new and the reemergence of existing but forgotten infectious diseases during the past 20 years. Flea-borne disease organisms are widely distributed throughout the world in endemic-disease foci, where components of the enzootic cycle are present. However, flea-borne diseases could reemerge in epidemic form because of changes in vector-host ecology due to environmental and human behavior modification. The changing ecology of murine typhus in southern California and Texas over the past 30 years is a good example of urban and suburban expansion affecting infectious disease outbreaks. In these areas, the classic rat-flea-rat cycle of R. typhi has been replaced by a peridomestic animal cycle involving, e.g., free-ranging cats, dogs, and opossums and their fleas. In addition to the vector-host components of the murine typhus cycle, we have uncovered a second typhuslike rickettsia, R. felis. This agent was identified from the blood of a hospitalized febrile patient and from opossums and their fleas. We reviewed the ecology of R. typhi and R. felis and present recent data relevant to the vector biology, immunology, and molecular characterization and phylogeny of flea-borne rickettsioses."} +{"text": "Tuberculosis: A Comprehensive Clinical Reference. H. Simon Schaaf, Alimuddin I. Zumla, John Grange, Mario Raviglione, Wing Wai-Yew, Peter Donald, Madhukar Pai, and Jeffrey Starke, 2009. London, UK: WB Saunders Elsevier \u00a3119, ISBN 9781416039884 (hardback) and ISBN 9788131221518 .Tuberculosis (TB) is today acknowledged as a global health catastrophe. In 1993, 111 years after Robert Koch's discovery of the cause of TB in 1882, the WHO declared TB \u201ca global emergency.\u201d In 2009, TB continues to kill 1.8 million every year and 5,000 people every day; that is one person every 20 seconds. Progress in TB management and control seems to have been lacking despite effective, inexpensive, and affordable chemotherapy being introduced over half a century ago. Over the past few years, interdisciplinary approaches and translational research on various aspects of TB is generating a large amount of data on conventional and molecular epidemiology, organism and host's interactions, new TB drugs and TB vaccine development, clinical management schedules for HIV-uninfected and HIV co-infected adults and children, development and evaluation of newer TB diagnostics, newer therapeutic regimens, and international standards of care for TB, among others. TB control programs around the world are faced with multidrug-resistant TB (MDR-TB) and extensively drug-resistant TB (XDR-TB), which are difficult and expensive to treat, threatening current efforts at TB control.Over the years, education and training for TB health care workers has been neglected and sidelined. Educational material for improving clinic staff knowledge to carry out current global WHO TB diagnostics, treatment, and management recommendations are lacking. These are crucial in detecting and eliminating active cases of TB who spread the disease in the community. Current clinical TB textbooks on TB are very expensive; unaffordable in developing countries; outdated; predominantly written by UK/US authors; and do not have updated recommendations for treatment of HIV-infected patients with TB. New information on management of TB generated from recent clinical and basic science research is not filtered through to the relevant TB health care providers at points of care where TB guideline booklets, formulated decades ago by the WHO, are still in use. There is a great need to update and educate these TB carers on the latest developments.For those of us with extensive experience in practicing medicine in a developing country, where poverty is rife and resources for TB care are scant, the quality of TB care delivered because of poor staff knowledge of the latest TB management recommendations hinders detection and effective treatment of active TB cases. The need for a comprehensive, up-to date textbook with the latest on TB management in HIV-infected and HIV-noninfected adults and children has been long overdue. A new TB textbook on the market has now fulfilled the needs described here. The textbook, entitled \u201cTuberculosis\u2014A Comprehensive Clinical Reference\u201d arises out of the global need to have a the latest developments in TB research and WHO-approved management guidelines incorporated into advice on day-to-day clinical practice for all TB care providers throughout the world. The editors of the textbook, global TB experts, Professors Zumla and Professor Simon Schaaf, teamed up with six international leading TB authorities as associate editors selected from across the globe: Professors John Grange (United Kingdom and Europe), Mario Raviglione , Wing Wai Yew (Hong Kong), Jeffrey Starke (United States), Madhukar Pai (Canada), and Peter Donald (South Africa). This editorial team used 156 eminent TB-experienced authors from several continents to write 107 chapters and 4 appendices constituting 1,008 pages to assemble this new textbook, a truly amazing and admirable achievement.MtB and HIV co-infection issues and gives practical day-to-day management guidelines for point of TB care internationally.This unique, extraordinary, and comprehensive book is mainly a clinical one with emphasis on practical application at points of TB care. It covers every aspect of the clinical, epidemiologic, immunologic, laboratory, management, social science, point-of-care delivery, and translational research aspects of TB that are highly relevant to clinical practice and TB control in developing and western countries. This book has been skillfully crafted by the editors for it to be applicable for use all over the globe and extensively covers both adult and pediatric TB with equal emphasis and equal proportions. The bulk of the chapters are on clinical presentation, diagnosis, management, and well-illustrated system-based case histories of adult and childhood TB. All 107 chapters are intelligible, easily readable, written to the highest quality, and cover up-to-date, evidence-based information on the state-of-the-art of a wide spectrum of important TB subject areas. TB affecting every organ and tissue in the body is comprehensively covered in detail. It also comprehensively identifies and deals effectively with In the short space provided here, it is difficult to do justice and highlight the superb features of this magnificent book. All chapters are highly illustrated (350 in the book), with elegantly presented figures, graphs, and tables adjunct to clinical, pathologic, and radiologic photos that show all minute details of the point being portrayed. The chapters review all current knowledge, gaps, and challenges ahead in various fields of TB. The book also deals with social and structural issues in TB and has very informative and useful appendices on conversion units for laboratory results, TB drug information, web sources on TB in general, and organizations supporting TB control and those providing research funding. This clinical textbook is aimed toward a variety of audiences, including general physicians, pulmonologists, TB and HIV clinic staff, medical assistants, nurse practitioners, TB program staff, medical students, postgraduates, technical staff, clinician scientists, policy makers, and donor agencies. The book opens with an emotional preface by His Grace, Archbishop Reverend Desmond Tutu, who himself had TB as a child. The book ends with a thought-provoking epilogue from Professors Zumla, Grange, and Schaaf, which introduces personal perspectives and optimistic caution.Importantly, to make this clinically based textbook widely available and affordable in developing countries, the editors' commitment to the TB cause and the publishers have simultaneously published a cheaper discounted edition for purchase in poorer developing countries. This price will be reduced further as charities such as Bookpower and donor agencies promote this invaluable textbook. No doubt this textbook is the best treatise on TB available on the market and will educate millions of health workers for a long time to come. This book is an excellent example of what can be achieved by uniting the scientific global community in the fight against TB."} +{"text": "With wide applications of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS), statistical comparison of serum peptide profiles and management of patients information play an important role in clinical studies, such as early diagnosis, personalized medicine and biomarker discovery. However, current available software tools mainly focused on data analysis rather than providing a flexible platform for both the management of patients information and mass spectrometry (MS) data analysis.Here we presented a plug-in-based software, BioSunMS, for both the management of patients information and serum peptide profiles-based statistical analysis. By integrating all functions into a user-friendly desktop application, BioSunMS provided a comprehensive solution for clinical researchers without any knowledge in programming, as well as a plug-in architecture platform with the possibility for developers to add or modify functions without need to recompile the entire application.http://sourceforge.net/projects/biosunms/.BioSunMS provides a plug-in-based solution for managing, analyzing, and sharing high volumes of MALDI-TOF or SELDI-TOF MS data. The software is freely distributed under GNU General Public License (GPL) and can be downloaded from With wide applications of MALDI-TOF MS and SELDI-TOF MS in biomedical studies, more and more large-scale MS datasets have being obtained -6. How tHere we developed a flexible and compact software, BioSunMS, for MALDI-TOF or SELDI-TOF MS-based clinical proteomics study. The name BioSunMS was coined by the combination of BioSun and MS (mass spectroemtry), in which BioSun stands for the comprehensive bioinformatics software developed by our center . BioSunMBioSunMS provides a relational database and client-server architecture suitable for multi-user workgroups. It is a RCP application extending the Eclipse framework. It was developed in Java, and provided a domain-specific platform where the plug-ins can be integrated. End-users can select related features from the BioSunMS plug-ins freely.BioSunMS system includes four functional modules, namely, data management, spectrum processing, MS profile analysis and security module Figure . The datBioSunMS is built using Standard Widget Toolkit (SWT). Created as part of the Eclipse project, SWT allows developers to build efficient and portable applications. In contrast to Swing/AWT , SWT has the same look and feel of the operating system on which the application runs. AWT/Swing components can be wrapped in SWT, and this feature is utilized in BioSunMS to integrate Java components built on these toolkits . In an ETo access the BioSunMS server at client side, users must first log on with an authorized username and a given password. Users could also log on for analyzing spectral data without connecting to the server, when their computers are offline. The workflow of BioSunMS was illustrated in Figure The first step in the workflow is to input patients general information and laboratory test results. To add a new record, click on the top-right button \"Append\" in the toolbar. A form will be displayed to enter data according to the module. Fields include equipment, sample ID, gender, age, specimen group and so on. Marked fields must be specified. Users could group samples with the Query View.The second step in the workflow is to import spectral files to the BioSunMS. After importing a spectral file, BioSunMS uses the Bioconductor PROcess package developed by Li to proceThe third step in the workflow is to group spectral files into the Bioresource folder. To analyze data, spectra must be grouped together into a folder. There are many ways to organize spectra by querying the database for spectra meeting desired criteria, such as research group, user, sample state, sample type, patient and characteristic description. Users can also define the condition by selecting a series of fields to be used to separate spectra into groups for classification and prediction. After grouping, a web page with a table of characteristics of patients can be created Figure .The fourth step in the workflow is to analyze the experimental results for identification of potential biomarkers and early diagnosis. BioSunMS provides the Wizard for general users, including extraction of MS peaks, construction of m/z matrix, sample classification and sample prediction. Advanced users can process and analyze the MS data in the Data Analysis Perspective. After building a Support Vector Machine (SVM) model on the training dataset and make prediction on the test dataset, BioSunMS uses a Receiver Operation Characteristic (ROC) curve to assess the models performance Figure .BioSunMS is a RCP application through bringing the power of lab's network to access and manage a large amount of information. It can be installed on Windows or Linux system. It allows researchers to enter patient data in a customizable template and group the data by queries. It has a security system for protecting data. Access is granted via user groups rather than individual users.BioSunMS is a network-based system like an Intranet. It can be accessed from all computers from the same local network. It can even be accessed through the internet. Therefore, data access and management can be password protected. The system always stores actions made by any users. The administrator can check the action history at any time.From the User and Group Management View, BioSunMS administrator can add or edit the user and the group. If a user group has been permitted to access a kind of data, any users belonging to the group are permitted to read the data in the Sample View. Any user who doesn't belong to the user group will not be permitted to read the data in the Sample View. Any user can create a new record, and modify a record which belongs to the same user group. However, only administrators can create new users and user groups. By default a new record is accessible to all users. To restrict access to a record, assign that record to the desired user group.Recording detailed information on collection, processing and storage of samples is crucial both for efficient reporting on biomedical study and for subsequent data analysis. Many paBioSunMS is useful for the users with the need to store, process and analyze MALDI-TOF or SELDI-TOF MS data. It supports general mass spectrometry file format, such as mzML, mzXML, mzData and CSV. For mzML data format standard released by the HUPO-PSI and Institute for Systems Biology in June 2008, BioSunMS reads and writes it using a package of ProteomeCommons.org IO Framework and ProtBioSunMS provides basic visualization tools and advanced processing algorithms. The method and related procedure, BioConductor PROcess was used for baseline subtraction, smoothing, normalization and peaks identification. Our data visualization module aims to support analysts in finding interesting peaks in spectra, selecting them for further analysis and visualizing the selected peaks by automatic or manual detection.The analysis of several spectra coming from biological samples belonging to different subjects focuses on identifying discriminant values in spectra related to diseases. The BioSunMS allows users to select features using t-test, construct models using SVM, and predict new samples by the wizard with the default parameters. Users can compare the generalization performance of a range of classifiers by plotting their performance on the test set in ROC curve. BioSunMS also provides heat maps and hierarchical clustering. In the future, we will gradually incorporate classification system Tclass and sampBioSunMS uses the R statistical programming language and some third part packages for processing and analysis of MS data. We implemented the communication between the BioSunMS and the R package with the class ncbaSpanker.System.R.RPackage. This class provides some methods for executing R scripts and commands. All communication with R language takes place via the file system. In general, BioSunMS writes a R script, makes the R system execute the script file, and output the results. Then BioSunMS reads an R output file to retrieve the results. The RPackage class hides all the details of communication with R. So users with little knowledge about R language can accomplish their task easily. Because many programs for gene expression profile-based analysis have been provided in R package, and some algorithms can be directly used in peptide profiling analysis, we use R package as the background computation. In addition, many packages for MS data analysis and visualization are also provided, for example, GALGO, caMassCHere we want to emphasize an important characteristic of BioSunMS. It can not only be used as the standard-alone program, but also can be a framework for developers. By taking full advantage of the editing and visualization components provided by Eclipse, the developers can focus entirely on the problems at hand. Moreover, BioSunMS was implemented in Eclipse platform, which made it more flexible and easier to adopt an algorithm as a plug-in. For example, we have successfully developed a plug-in, cn.org.biosun.knn, for sample classification using k-nearest neighbour (kNN) method.BioSunMS is a plug-in based and flexible software for MS data management and analysis. It integrates patients information and MS data storage, process, sample classification and sample prediction into a single, user-friendly workbench. The project provides an alternative solution to analyze high throughput MS data from MALDI-TOF or SELDI-TOF.The future for BioSunMS holds much potential with many plug-ins in development, including Web service, more statistical methods and machine learning algorithms, such as GA, Decision-Tree, Na\u00efve Bayes method, and sample class discovery. There is also ongoing work for improving the plug-ins which allows researchers to enter patients data in a structured way using a customizable domain model. Another major feature in development is online updates for plug-in from the BioSunMS update server. The current status of the BioSunMS development can be viewed at the BioSunMS website.Project name: BioSunMSProject home page: http://ccb.bmi.ac.cn/biosunms/ or http://sourceforge.net/projects/biosunms/Operating system(s): Windows, LinuxProgramming language: Java, R, SQLOther requirements: SUN JVM 1.5 or higher, R Package 2.5.1 or higher, MySql 5.0 or higherLicense: GNU GPLAny restrictions to use by non-academics: Contact authorsRCP: Rich Client Platform; SWT: Standard Widget Toolkit; MS: Mass Spectrometry; MALDI-TOF MS: Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry; SELDI-TOF MS: Surface-Enhanced Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry; ROC: Receiver Operation Characteristic; SVM: Supported Vector Machines; kNN: k-Nearest NeighbourThe authors declare that they have no competing interests.YC designed and developed the system. XY, NW, AL and HW did the conceptual design of the system and gave some detailed technique supports. XZ and WL conceived the project, performed overall supervision and coordination. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1472-6947/9/13/prepub"} +{"text": "Previous population-based studies showed differences in international and within country colorectal cancer survival estimates, but few investigated the role of prognostic factors. Using a \"high resolution approach\", we aimed to determine the effect of ethnicity and health care by comparing Filipino-Americans with Philippine residents, who have the same ethnicity, and with Caucasians living in the US, who have the same health care system.Using databases from the Manila and Rizal Cancer Registries and the United States Surveillance, Epidemiology and End Results, age-adjusted five-year absolute and relative survival estimates were computed and compared between Filipino-American colorectal cancer patients, cancer patients from the Philippines and Caucasian patients. Cox proportional hazards modelling was used to determine factors affecting survival differences.Much lower 5-year relative survival estimates were obtained for Philippine residents (37%) as compared to those in Filipino-Americans (60.3%) and Caucasians (62.4%). Differences in age, stage and receipt of surgery explained a large proportion of the survival differences between Philippine residents and Filipino-Americans. However, strong excess risk of death for Philippine residents remained after controlling for these and other variables .Strong survival disadvantages of Philippine residents compared to Filipino-American patients were disclosed, which most likely reflect differences in access to and utilization of health care. Health education and advocacy, for both patients and health practitioners, should likewise be given priority. Colorectal cancer (CRC) is among the most common cancers worldwide, ranking fourth in men and third in women . IncidenWithin countries, incidence, mortality and survival rates were reported to vary. In the United States, incidence and mortality among Caucasians were lower than among African-Americans, but higher than among Asian and Pacific Islanders (API) and Hispanics . Five-yeWhile international and interracial CRC survival comparisons have been reported, high resolution studies that could elucidate the role of prognostic factors, such as stage and treatment, are rare ,4,5,7. TIn previous studies, variations in access to diagnostic and treatment facilities were put forward as the reasons for survival differences between countries and populations ,4,7. HowFurthermore, there is also a dearth of published population-based colorectal cancer survival estimates from developing countries, and comparisons of these with data from developed countries are limited ,10,11. SFrom the Philippines, population-based cancer survival data were reported for the first time in the 1998 International Agency for Research on Cancer (IARC) monograph \"Cancer Survival in Developing Countries\" , and onlTo elucidate the role of various factors, including ethnicity, stage at diagnosis, and access to treatment in CRC survival, we compared Filipino-Americans with Philippine residents, who have the same ethnicity, and with Caucasians living in the US, who are exposed to similar health care systems.Using the United States Surveillance, Epidemiology and End Results (SEER) 13 database , CRC patInformation on CRC cases who were residents of the National Capital Region (NCR) were abstracted from the databases of the Philippine Cancer Society-Manila Cancer Registry (PCS-MCR) and the Department of Health-Rizal Cancer Registry (DOH-RCR). The PCS-MCR covers the four major cities of the NCR , while the DOH-RCR covers the 12 municipalities of the former Rizal province that were incorporated into the NCR and the present Rizal Province. The NCR holds the seat of the Philippine government and is the largest urban metropolis, and the political, social, educational and economic center.Data collection procedures include exhaustive identification and collection of information on patients from all hospitals in the NCR and the Rizal Province, which limits underreporting to a minimum. Various patient records were reviewed, including medical, pathology, radiotherapy, radiology, ultrasound, nuclear medicine and CT scan reports, and records from hospital tumor registries, if available. Death certificate notifications (DCN) indicating cancer as the cause of death are also routinely acquired by the registries from all the Local Civil Registry Offices in the constituent cities.The Philippine registries are regarded as among the high-quality cancer registries from developing countries and have consistently been included in Cancer Incidence in Five Continents -20. TheyThe same inclusion and exclusion criteria as for the SEER databases were used in the subject selection. For the Philippine data though, due to limited resources, only subsamples of 200 cases diagnosed in each calendar year from 1993 to 2002 were randomly selected from the 7,769 eligible CRC patients in the database. These were followed with respect to vital status until December 31, 2002 and included in the analysis. From DCNs mentioning cancer as the cause of death, survival status was assessed. Active follow-up by personal visits to the patients or their families in the last known place of residence was used to confirm status for those not identified as dead.From the 2000 randomly sampled patients, 160 (8%) were excluded from the analysis due to invalid data while an additional 205 (10.2%) were removed due to the absence of any follow-up information. Anonymized datasets for the remaining 1,635 colorectal cancer patients (81.8%) were prepared and used in the analysis. Of these, 83.6% have complete follow-up information. Of the patients who are alive, 68.6% have complete follow-up information. For survival analyses, patients with incomplete follow-up information were censored at the last date known alive.From the patients who have complete follow-up information, 53.9% were identified to be deceased, of whom 98.2% have cancer as the cause of death. For 82.3% of patients who are dead, information on the cause of death was obtained from medical records or death certificates, while data for another 12.6% were acquired through interviews of family members. Survival information for only a few (5.1%) came from other sources.The project proposal was approved by the Ethics Review Board of the National Institutes of Health of the University of the Philippines Manila.Conventionally, cohort-based analyses, using the life-table method or the Kaplan-Meier method ,22, haveAs commonly practiced in population-based survival analysis, estimates of relative survival are reported, in addition to estimates of absolute survival. Expected survival for the general population of analogous age distribution was derived using the so-called Ederer II method and lifeTo test for differences in survival between the three cancer populations, a novel modelling approach for period analysis was usedTo explain possible survival differences and to determine factors affecting survival, both within and between the three cancer patient populations, the Cox Proportional Hazards model was used. Initially, separate Cox models by population group were used to determine bivariate associations of age, sex, stage, histology, surgery and radiotherapy with survival. A multivariate model was then built jointly for all three groups to compare survival probabilities between populations. Relative risks were calculated using Filipino-Americans as the reference group, while controlling for the effects of age, sex, stage, histology, surgery and radiotherapy, first individually and then simultaneously. Those with missing information were excluded in the multivariate analysis. For each variable included in the Cox models, the proportional hazards assumption was checked by using log (-log) graphs. The plotted lines were roughly parallel over time and no violations of the proportional hazards assumption were found.Age at diagnosis was categorized into the 5 age adjustment groupings mentioned earlier. Stage categories were localized, regional and distant, while histology was classified based on the WHO Classification of Tumours . It is unlikely that patients lost to follow-up have higher survival than those who were not, since it can be assumed that they have more advanced diseases. Furthermore, it can be surmised that they have not received any form of treatment, as follow-up data for these patients were not found in hospitals within the NCR, where majority of specialized cancer care facilities are located, and the availability of cancer treatment is limited in the surrounding areas. Given these circumstances, it can be deduced that the survival estimates are higher than what could be expected, should follow-up be complete for all patients.In conclusion, the differences in CRC survival between Filipinos and Filipino-Americans underscore the importance of access to health care. Improving access to and utilization of diagnostic and treatment facilities and making them affordable in the Philippines should be emphasized. Likewise, the important role of health education and advocacy, particularly in promoting early diagnosis and prompt treatment should be given priority, for both patients and health practitioners. The comparison between Filipino-Americans and Caucasians indicates that CRC survival can be similar in different ethnic groups, given comparable access to health care.API: Asian and Pacific Islanders; AS: Absolute survival; CI: Confidence intervals; CRC: Colorectal cancer; DCO: Death certificates only; DCN: Death certificate notifications; DOH-RCR: Department of Health-Rizal Cancer Registry; RR: Relative risk; IACR: International Association of Cancer Registries; IARC: International Agency for Research on Cancer; ICD-O: International Classification of Diseases for Oncology; NCR: National Capital Region; NHW: non-Hispanic White; PCS-MCR: Philippine Cancer Society-Manila Cancer Registry; RS: Relative survival; SE: Standard error; SEER: Surveillance, Epidemiology and End Results; WSCPP: World Standard Cancer Patient Population; US: United States.The authors declare that they have no competing interests.The contributions of the authors are as follows: MTR contributed in the planning of the study, supervised data collection, performed the analysis and wrote the manuscript; AL, MRL and GU planned and supervised data collection, reviewed and edited registry abstracts, and performed data management; AG assisted in the analysis; HB contributed in the planning of the study and supervised data analysis and writing of the manuscript. All authors have read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2407/10/100/prepub"} +{"text": "The Author Contributions section incorrectly assigns a single member of the SEA-ORCHID Study Group to the study\u2019s design, conduct, analysis, interpretation, and reporting. The correct contribution of each member of the SEA-ORCHID Study Group is listed in the Acknowledgments section."} +{"text": "Entamoeba histolytica is a pathogenic ameba that has recently been recognized as an emerging pathogen in men who have sex with men (MSM) in Asia-Pacific countries where it is not endemic, i.e., Japan, Taiwan, and Republic of Korea. We report locally acquired invasive amebiasis in Sydney, Australia, exclusively in MSM. Entamoeba histolytica is a pathogenic ameba that can cause invasive intestinal and extra-intestinal disease. The most frequent manifestations of invasive amebiasis are colitis and liver abscesses , and median CD4 count was 713 cells/mm3 . No assoE. histolytica/dispar/moshkovskii complex. Diagnosis of E. histolytica was confirmed by PCR targeting the small subunit ribosomal DNA as described . In 1 patient, the abscess ruptured through the liver capsule, and collapse of the right middle and lower lung lobes and a resultant pleural effusion complicated the subphrenic collection of pus from the abscess. Levels of liver enzymes , C-reactive protein, and neutrophils (absolute numbers) were raised. Both patients underwent percutaneous drainage of their liver abscess; cultures of aspirated pus were negative for bacteria and fungi. Results of indirect hemagglutination antibody assay were positive; titer was All 5 patients made a successful recovery after treatment with metronidazole. In addition, all were treated for cyst carriage with paromomycin, a luminal amebicide.E. histolytica carriage and invasive disease are common in the Asia-Pacific region, especially in developing countries. In countries where E. histolytica prevalence is low, such as Japan, Taiwan, Republic of Korea, and Australia, rates of amebiasis are low and invasive amebiasis is uncommon. Recent reports from a number of these countries, however, suggest that invasive amebiasis is emerging as an increasingly common infection, specifically in the MSM population are higher for HIV-infected persons then for HIV-nonninfected persons, although the reasons are unclear (E. histolytica carriage in MSM likely reflect high-risk sex behavior and multiple exposures, resulting in increased risk for acquisition. This hypothesis is supported by the high rates of sexually transmitted infections that occur in Australian MSM who visit sex-on-premises venues (Of note, 4 of the 5 patients we report were HIV infected. Seroprevalence rates of E. histolytica in MSM is of public health concern because it has the potential to become endemic in this population in Australia and to cause severe disease. Further study is needed to identify the reasons for the geographic variation and the role of E. histolytica in invasive disease. In conclusion, invasive amebiasis has the potential to emerge as an important parasitic infection in the Asia-Pacific region, especially in HIV-infected MSM in countries where E. histolytica is not endemic.The emergence of"} +{"text": "Empirical evidence demonstrates that informal patient payments are an important feature of many health care systems. However, the study of these payments is a challenging task because of their potentially illegal and sensitive nature. The aim of this paper is to provide a systematic review and analysis of key methodological difficulties in measuring informal patient payments.The systematic review was based on the following eligibility criteria: English language publications that reported on empirical studies measuring informal patient payments. There were no limitations with regard to the year of publication. The content of the publications was analysed qualitatively and the results were organised in the form of tables. Data sources were Econlit, Econpapers, Medline, PubMed, ScienceDirect, SocINDEX.Informal payments for health care services are most often investigated in studies involving patients or the general public, but providers and officials are also sample units in some studies. The majority of the studies apply a single mode of data collection that involves either face-to-face interviews or group discussions.One of the main methodological difficulties reported in the publication concerns the inability of some respondents to distinguish between official and unofficial payments. Another complication is associated with the refusal of some respondents to answer questions on informal patient payments.We do not exclude the possibility that we have missed studies that reported in non-English language journals as well as very recent studies that are not yet published.Given the recent evidence from research on survey methods, a self-administrated questionnaire during a face-to-face interview could be a suitable mode of collecting sensitive data, such as data on informal patient payments. Informal patient payments are an important feature of health care systems in many countries around the world ,2. In soDespite the different explanations, informal patient payments are overall seen as a negative feature of health care provision. Informal patient payments can have adverse effects on equity and can hinder the determination of future funding requirements of the health care sector ,6-8. EmpAlthough the importance of data on informal patient payments is universally recognised, the collection of such data is a challenging task given their informal and potentially sensitive nature -14. MoreMethodological difficulties in collecting sensitive data are an important topic in research on survey methods e.g. -19]. In . In 19]]The aim of this paper is to critically review the research designs applied to the investigation of informal patient payments following the method of a systematic literature review. Our review is expected to facilitate the development of future research designs for collecting valid and reliable data on informal patient payments. Such critical review has not yet been reported in the literature. To achieve our aim, we first define the term \"informal patient payments\". Based on this definition, we identify keywords to search systematically for relevant publications. The following sections present our definition of informal patient payments and the methods of data collection, followed by the results and their discussion.Empirical studies on informal patient payments attribute different characteristics to this type of payments. As a result, informal patient payments do not have a universal definition although the definitions used by researchers partly overlap.For example, Adam who repoOther recent studies add the moment of payment to the definition of informal patient payments. According to Allin et al \"informaDespite the difference in the definitions, it is generally accepted that informal patient payments could have monetary and non-monetary form, and could express the patient's gratitude but could also be requested by the health care provider. Overall, informal patient payments are accepted to be unofficial, i.e. they are not registered by the state and are made without an official receipt of payment, and remain outside the official payment channels. However, confusion arises when these payments are also defined as illegal. This is because informal payments are sometimes - but not always - illegal . These pBased on the definitions of informal patient payments discussed above and reported in other publications e.g. ,30], it , it 30]]\u25aa Who initiates the informal payment? The patient who wishes to express gratitude, the provider who requests the payment, or both?\u25aa What is the nature of informal payment? In cash, in kind , or in a form of services ?\u25aa What is the moment of informal payment? Before, during, or after the health care service, medical supplies or pharmaceuticals are provided to the patient?\u25aa Who receives the informal payment? The health care institution , medical staff (incl. physicians and nurses), or the administration of the health care institution?\u25aa Who actually makes the informal payment? The patient or the relatives of the patient?\u25aa What is the purpose of informal payment? Expression-of-gratitude, fee-for-service, fee-for-commodity, fee-for-access, fee-for-quicker-access, or fee-for-better-quality?\u25aa What is the amount of the informal payment? The monetary value of the informal patient payment is usually compared to the household's income.\u25aa How is the informal payment perceived? Normal behaviour, corruption, illegal behaviour, or tradition ?\u25aa What is the attitude toward the informal payment? Negative or positive (if an expression of gratuity), usually depending on the moment of payment?The characteristics of informal patient payments presented above, cannot be analysed separately because they may correlate among each other or one characteristic may even be a cause of another characteristic. For example, when the payment is requested, it is usually observed as a cash payment to medical staff in a surgical department, and the amount of the payment can be higher than the monthly income of the patient. At the same time, a gratuity payment that is in kind has a value that corresponds to the patient's income. Despite the possibility of such correlations, to be able to understand the phenomenon of informal patient payments, all characteristics listed above should be taken into account.We take these characteristics as a definition of informal patient payments for our analysis. However, we focus solely on informal patient payments for health care services excluding informal patient payments for medical supplies and pharmaceuticals.In order to indentify the research techniques used in the study on informal patient payments for health care services, we conducted a systematic literature review using the method of desk research. The combinations of keywords used for the search of relevant literature, consisted of two components. The first component contained the term \"informal patient payments\" or one of its synonyms (see previous section), namely \"unofficial out-of-pocket payments\", \"under-the-counter payments\", \"under-the-table payments\", \"envelop payments\", and \"corruption in health care\". The second component consisted of the term \"empirical research\" or one of its synonyms, namely \"survey\" and \"study\".Using all possible combinations of keywords in each of the two components, the following databases were searched: Econlit, Econpapers, Medline, PubMed, ScienceDirect, SocINDEX. Only English language publications were selected for further analysis. There were no limitations with regard to the year of publication or publication status. Each publication identified in the systematic search for literature, was checked for its relevance with regard to our research questions. Only publications that reported on empirical studies were included in the list of relevant publications. If it was obvious that the same empirical study was reported in more than one publication, only one publication was included in the final list but all publications that reported the study, were taken into account to identify details related to the study design. We also reviewed the reference lists of the publications that we identified for other relevant studies.The content of the publications was analysed qualitatively and the results were organised in the form of tables. The main objective of the analysis was to outline the research designs reported in these publications. The focus was to extract information on the data collection process (i.e. sample characteristics and data collection mode) and the research instrument . The research results reported in the studies were also summarised (using the definition outlined in the previous section) to indicate the type and incidents of informal payments for health care services reported in the literature. The results reported in the tables, were assessed in view of findings reported in the literature on research methods.In total, 31 publications were identified as relevant in the systematic literature review. The publications are presented in Appendix 1 according to the year of their publication starting from the most recent ones. This section presents the general study description, data collection process, research instruments, as well as types and incidents of informal patient payments reported in these publications.The general description of the studies included in our review is presented in Table Research on informal payments for health care services was conducted in virtually all continents. This includes countries with low-, lower-middle-, upper-middle- and high-income economies . MoreovThe specificities of the data collection process reported in the publications that we reviewed, are presented in Table Of all studies included in our review, 18 publications reported one type of data collection mode. In the case of consumers, the most frequently used mode of data collection was face-to-face interview. In the case of providers, face-to-face interviews were also widely used, but besides them, focus-groups interviews and self-administrated questionnaires were also used to gather data. Focus-group discussions and questionnaires were applied to consumers as well. Overall, self-administrated questionnaires were seldom used as a research instrument. Few publications reported a mixing mode of data collection combining interviews and group discussions. Respondents were not the only source of data. For example, one article provided content-analysis of printed media.Additional analysis suggested that the collection of data on informal patient payments had changed over the years. At the beginning, only the general public and providers were involved in the studies, while later, patients and official were also included as sampling units. The number of sampling areas and sample selection techniques applied in the studies increased over the years. Thus, researchers included not only probabilistic sample designs but also purposive, snowball and convenience samples in the recent years, as well as larger sampling areas. The samples in some recent studies were very large (more than 10000 units) compared to earlier studies. The data collection had also become more varied with the years including more types of data collection modes. An interesting example is the stakeholder workshop applied in one recent study.We also considered the response rate reported in the publications but we found that only 9 out of 31 publications indicated this feature of data collection. When reported, the response rate was quite high ranging form 70% to higher than 90%. Only one study based on telephone interviews reported a response rate lower than 20%. Nevertheless, the limited number of publications that report the response rate precludes a meaningful comparison in this direction.Table Although, we could identify groups of questions on informal payments for health care services included in the studies, overall, the content of the research instrument was rarely described in detail. The piloting and pre-testing were also not described in details in any of the publications.With regard to the recall period, researchers frequently appealed to the memory of respondents when the experience with paying informally was the objective of the survey. There were only two options of the recall period applied to the providers and officials: last week and two years ago. However, we found a variety of recall periods applied in studies among consumers. Respondents were asked to remember making payments during a year or more, as well as during one to five months. Next visit and last visit were also used as reference points in the studies.Two publications stated that the questionnaire was pre-tested and 10 publications provided information that the questionnaire was piloted. In cross-national studies, a backward translation was usually applied to ensure the proper wording of the questions. The introduction of country specific questions was also used in these studies.Tables Researchers reported a variety of beneficiaries of informal payments for health care services, e.g. general practitioners, medical specialists, other medical staff, and administration. Overall, respondents reported higher informal payments for services of medical specialists (notably surgeon and dentist) than for services of general practitioner although we observed that researchers appealed more often to informal payments to general practitioners. The expression of gratitude was identified as a motivation for informal patient payments in about a quarter of the studies while more than a quarter of the studies reported the improved service provision as the main reason for such payments.The magnitude of informal patient payments was rarely reported (only in 5 publications). Nevertheless, this characteristic of informal patient payments was hardly comparable since researchers were using different measurement units: monthly household income, or monthly household expenditure, or health expenditures.The few studies that investigated the perception and attitude of respondents toward informal patient payments, reported quite contrasting results. Informal patient payments were perceived by respondents as tradition and gratuity in 3 studies, and in other 3 studies they were perceived as illegal behaviour and corruption.The incidence of informal patient payments reported in the publications that we reviewed . Therefore, a universal definition is not available. The key characteristics described at the outset of this paper provide a more appropriate base for studying this phenomenon than pursuing an all-inclusive definition. Still, country-specific features should be taken into account to make sure that the unit used to measure informal payments is meaningful to the population being sampled.The results of our review suggest that the study of informal patient payments for health care services is rather new, though the phenomenon has been in existence for a number of decades . Most ofOur findings confirm that informal payments exist in countries of all levels of economic development, and in different parts of the world. However, we did not find studies reporting informal patient payments in high-income countries in North-West Europe, North America and Australia. The phenomenon is most often observed in former-socialist countries and developing countries , although it also exists in some high-income European countries that were not former-socialist countries ,36. As mWhen we look at the study designs that we reviewed, we can outline several discussion points relevant to research. The first discussion point refers to the study objectives. We differentiated between exploratory, descriptive, analytical and predictive aims. However, we did not find studies with an explicit exploratory aim even among the earlier studies. We expected that an exploratory aim would be typical for the early studies when scant information was available because then, the research interest would be concentrated on exploring the phenomenon. Although some earlier studies had an explicit descriptive aim, other earlier studies had an analytical aim. Descriptive and analytical objectives allow finding determinants of informal patient payments and their correlation.The second discussion point refers to the sample design. Sample design is part of the entire research design and it may minimise some biases in case of a well-developed sample. To estimate the level of informal patient payments, a probabilistic sample strategy is commonly implemented as it gives equal chances of being included in the study. Moreover, triangulation of the data is feasible when all parties participate in the research. For instance, Cockroft et al present Another discussion point is the data collection mode. The mode of data collection can be especially problematic when sensitive data are studied. This is because each single mode of data collection has its own pros and cons when sensitive questions are asked. The mode of data collection might even be a determinant of the value of indictors estimated based on sensitive data e.g. ,19]. Acc. Acc19]]Virtually all publications that we reviewed are based on retrospective research, thus another relevant discussion point is the recall period. The human memory can be a source of bias in research . ConsumeOur findings on the response rate were surprising to a certain extent. In general, the response rate reported in the publications that we reviewed, was rather high. This could suggest that people are willing to talk about informal patient payments despite their informal and potentially illegal nature. However, it should be recognised that only few publications presented this characteristic. It might be that the response rate was presented in these publications because it was favourable for the study and indicated the representativeness of the data.To enrich the methodological approaches to the investigation of informal patient payments, researchers can appeal to methods for measuring corruption in society. Although informal patients are not always illegal, our review suggests that they are sometimes perceived by respondents as corruption and illegal behaviour. The literature on measuring corruption suggests that corruption can be studied through the measurement of perceived corruption, as well as perceived willingness to pay bribes and bribe payments . SpecifiThe key results on the type of informal patient payments indicate that informal patient payments are a multifaceted phenomenon. All characteristics of informal patient payments included in our definition, appeared relevant for describing the pattern and magnitude of informal payments for health care services. Overall, the results indicate a great variety in the types of informal patient payments reported. This needs to be considered when designing a research instrument for the investigation of these payments. In particular, the researcher needs to clarify in advance what types of informal patient payments should be studied and thus, what type of questions to be included. It is also important to decide how to measure the incidents of informal patient payments since various measurement units are possible.Our attempt to compare the empirical results presented a significant challenge. This is mainly due to the great variety of research methods applied. However, the overall findings indicate that informal patient payments are a substantial phenomenon in terms of both scope and scale, and should not be neglected. Moreover, results of household surveys would be more meaningful if considered against the background of macro-level data at a national level (whenever available). For example, the National Health Accounts could be a useful source of macro-level data since they report total health expenditures as well as formal transactions in the health care sector . In addition to this, little is known on why informal patient payments exist and how the specific patient-providers relationship determines them. This indicates the need of combining quantitative and qualitative research methods when studying this type of payments. The need of deeper understanding of the informal patient payments has already captured the attention of researchers who are trying to provide theoretical explanations to the existing empirical findings ,44.We searched systematically for relevant publications. However, we do not exclude the possibility that we have missed some studies reported in non-English language journals as well as very recent studies that are still not reported. Despite this shortcoming, our results and discussion are relevant to future research on informal patient payments. As mentioned above, the investigation of the phenomenon is interwoven with methodological complexities related primarily to the data collection and research instruments. We have outlined and discussed most of these complexities. However, other peculiarities (e.g. wording of the questions and the length of the interview) also require attention.Based on our findings in combination with the conclusions of a recent methodological review presented in Roberts , the folPredictors of informal health payments: The example from Turkey. Journal of Medical Systems 2010, 34(3): 387-396.1. Ozgen H, Sahin B, Belli P, Tatar M, Berman P: An inter-country comparison of unofficial payments: Results of a health sector social audit in the Baltic States. BMC Health Services Research 2008, 8:15.2. Cockcroft A, Andersson N, Paredes-Solis S, Caldwell D, Mitchell S, Milne D, Merhi S, Roche M, Konceviciute E, Ledogar R: Informal payments in public hospitals in Greece. Health Policy 2008, 87:72-81.3. Liaropoulos L, Siskou, O, Kaitelidou D, Theodorou M, Katostaras T: Private health expenditure in the Greek health care system: Where truth ends and the myth begins. Health Policy 2008, 88:282-293.4. Siskou O, Kaitelidou D, Papakonstantinou V, Liaropoulos L: Access to medicines and out of pocket payments for primary care: Evidence from family medicine users in rural Tajikistan. BMC Health Services Research 2008, 8:109.5. Tediosi F, Aye R, Ibodova S, Thompson R, Wyss K: Informal payments and moonlighting in Tajikistan's health sector. Washington, D.C.: The World Bank and Development Research Group; 2008.6. Dabalen A, Wane W: Journal of Applied Social Psychology 2007, 37: 1060-1076.7. Burak LJ, Vian T: Examining and predicting under-the-table payments for health care in Albania: An application of the theory of planned behavior. Gifts, bribes and solicitions: Print media and the social construction of informal payments to doctors in Taiwan. Social Science & Medicine 2007, 64:521-530.8. Chiu YC, Smith KC, Morlock L, Wissow L: Bribery in Health Care in Peru and Uganda. NBER Working Paper. Cambridge: NBER; 2007. http://www.nber.org/papers/w130349. Hunt J: Informal payments in the health sector: A case study from Turkey. Health Affairs 2007, 26:1029-1039.10. Tatar M, Ozgen H, Sahin B, Belli P, Berman P: Central Asian Survey 2006, 25:441-460.11. Baschieri A, Falkingham J: Formalizing informal payments: The progress of health reform in Kyrgyzstan. The inequity of informal payments for health care: The case of Hungary. Health Policy 2006, 75:262-271.12. Szende A, Culyer AJ: Informal payments in government health facilities in Albania: Results of a qualitative study. Social Science & Medicine 2006, 62:877-887.13. Vian T, Grybosk K, Sinoimeri Z, Hall R: Health care-seeking behaviour and out-of-pocket payments in Tbilisi, Georgia. Health Policy and Planning 2005, 20:232-242.14. Gotsadze G, Bennett S, Ranson K, Gzirishvili D: Out-of-pocket payments and utilization of health care services in Albania: Evidence from three districts. Health Policy 2005, 75:18-39.15. Hotchkiss DR, Hutchinson PL, Malaj A, Berruti AA: Health service utilization in the Former Soviet Union: Evidence from eight countries. Health Services Research 2004, 39:1927-1950.16. Balabanova D, McKee M, Pomerleau J, Rose R, Haerpfer C: Out-of-pocket and informal payments in health sector: Evidence from Georgia. Health Policy 2004, 70:109-123.17. Belli P, Gotsadze G, Shahriari H: Poverty, out-of-pocket payments and access to health care: Evidence from Tajikistan. Social Science & Medicine 2004, 58:247-258.18. Falkingham J: Informal Out-of-Pocket Payments for Healthcare in Russia. Independent Institute for Social Policy: Moscow; 2003.19. Shishkin S, Bogatova T, Potapchik Y, Chernets V, Chirikova A, Shilova L: Health Policy 2002, 62:243-273.20. Balabanova D, McKee M: Understanding informal payments for health care: The example of Bulgaria. Formal and informal household spending on health: a multi-country study in central and eastern Europe. Cambridge, MA, Harvard School of Public Health; 2003. Belli, 2002.21. Belli P: Underground Economy in Health Care in Contemporary Ukraine. Odesa: TEC; 2001.22. Litvak A, Pogorilyi V, Tyschuk M: Institutional Issues in Informal Health Payments in Poland: Report on the Qualitative Part of the Study. HNP Discussion Paper. Washington, D.C.: The World Bank; 2001.23. Shahriari H, Belli P, Lewis M: Corruption in Slovakia: Results of diagnostic surveys. Washington, D.C.: The World Bank and USAID; 2000.24. Anderson J: 'If you pay, we'll operate immediately'. Journal of Medical Ethics 2000, 26:305-311.25. Miller WL., Grodeland, AB, Koshechkina TY: Unofficial fees in Bangladesh: Price, equity and institutional issues. Health Policy and Planning 1999, 14:152-163.26. Killingsworth JR: Out-of-pocket payments for health care in Croatia: Implications for equity. Croatian Medical Journal 1999, 40:152-159.27. Mastilica M, Bozikov J: Informal economic activities of public health workers in Uganda: Implications for quality and accessibility of care. Social Science & Medicine 1999, 49:849-865.28. McPake B, Asiimwe D, Mwesigye F, Ofumbi M, Orteublad L, Stree P: Financing health services in Poland: New evidence on private expenditures. Health Economics 1998, 7:337-346.29. Chawla M, Berman P, Kawiorska D: Under-the-counter payments for health care: Evidence from Bulgaria. Health Policy 1997, 42:89-100.30. Delcheva E, Balabanova D, McKee M: Journal of Medical Ethics 1996, 22:33-40.31. Barr DA: The ethics of Soviet medical practice: Behaviours and attitudes of physicians in Soviet Estonia. The authors declare that they have no competing interests.TS carried out the searching of the literature, drafted the results tables and manuscripts; MP developed the design of the study and a conception for data interpretation; improved the draft; IG participated in the sequence alignment, revised the paper; WG participated in the sequence alignment; revised the paper. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1472-6963/10/273/prepub"} +{"text": "It is important to find a comorbidity measure with better performance for use with administrative data. The new method proposed by Elixhauser et al. has never been validated and compared to the widely used Charlson method in the Asia region. The objective of this study was to compare the performance of three comorbidity measures using information from different data periods in predicting short- and long-term mortality among patients with acute myocardial infarction (AMI) and chronic obstructive pulmonary disease (COPD).We conducted a retrospective cohort study using National Health Insurance claims data (2001-2002) in Taiwan. We constructed the Elixhauser, the Charlson/Deyo, and the Charlson/Romano methods based on the International Classification of Disease, 9th Revision, Clinical Modification codes in the claims data. Two data periods, including the index hospitalization as well as the index and prior 1-year hospitalizations, were used in the analysis. The performances were compared using the c-statistics derived from multiple logistic regression models that included age, gender, race, and whether the patient received surgery or not. The outcomes of interest were in-hospital and 1-year mortality.The performance was in the same rank order among both populations regardless of the outcome and data period: Elixhauser > Charlson/Romano > Charlson/Deyo. In predicting in-hospital mortality, the Elixhauser models using information from the index hospitalization performed best, even better than the Charlson/Deyo or Charlson/Romano models using information from the index and prior hospitalizations. Nevertheless, in predicting 1-year mortality, the Elixhauser models using information from the index and 1-year prior hospitalizations performed better than using information from the index hospitalization only.This is so far the first study to validate the Elixhauser method and compare it to other methods in the Asia region, and is the first to report its differences in data periods between short- and long-term outcomes. The comorbidity measurement developed by Elixhauser et al. has relatively good predictive validity, and researchers should consider its use in claims-based studies. Administrative databases are increasingly used in health services research, epidemiologic research, and outcome studies. For control of baseline differences in these observational data, one major issue is to find a comorbidity measure with better performance -9. Evideth Revision, Clinical Modification (ICD-9-CM) codes in administrative databases were made by Deyo et al.[Among various methods, the Charlson comorbidity index (CCI) has been used most widely. It was developed by considering the impact of comorbidities in predicting 1-year mortality of medical inpatients using comorbidity data recorded in the medical charts . Two adayo et al. , and thus is more complicated to use. The Elixhauser method has more variables than the Charlson based methods, and therefore needs larger sample size. However, the increasingly used administrative databases make sample size large enough for analysis.Factors influencing the performance of claims-based comorbidity measures include the dataset, studied population, outcome, and data periods ,19. CompData are collected from the National Health Insurance inpatient claims data, family registration file, and death certification data. An encrypted unique identification number is used to link information of the same patient from different datasets.http://www.nhi.gov.tw/; http://w3.nhri.org.tw/nhird//index.php). The inpatient claims data include the dates of admission and discharge, sex, birth date, and diagnosis and procedure codes using the International Classification of Disease, 9th revision, Clinical Modification (ICD-9-CM). Each inpatient record includes the principle and up to 4 secondary diagnoses as well as 3 procedures.Taiwan had a universal single-payer National Health Insurance Program since 1995. As of 2002, 97.1% of Taiwan's population was enrolled in this program [http://www.ris.gov.tw/). The database of death certificates, managed by the Department of Health in Taiwan, is a national registry of all deaths in Taiwan (http://www.doh.gov.tw/).The family registration file is used to identify the race of the patients . This database is maintained by the government and provides relatively accurate demographic information on residents (ICD-9-CM: 410.x) and chronic obstructive pulmonary disease (COPD) . We used the Clinical Classifications Software (CCS), which is published and freely downloadable from the website of the Agency for Healthcare Research and Quality (AHRQ) (.ahrq.gov), to defThere were two data periods used in our analysis. One is the index hospitalization. The other one is the index and prior 1-year hospitalizations. The index and prior 1-year hospitalizations had 1-year lookback period. The index hospitalization didn't have lookback period. The index hospitalization was identified as the first hospitalization of our study population during year 2002. The index and prior 1-year hospitalizations included the index hospitalization and all hospitalizations 1-year before the index date.In-hospital all-cause mortality and 1-year all-cause mortality were selected because they represented short- and long-term outcomes. The administrative claims data was linked to death certification data to identify when the patient died.We compared three published claims-based comorbidity measures ,13,18. Thttp://www.ahcpr.gov/data/hcup/comorbid.htm[The third comorbidity method is developed by Elixhauser et al. using administrative data from California . We idenorbid.htm), and weorbid.htm.Two data periods, including the index hospitalization as well as the index and prior 1-year hospitalizations, were used to predict in-hospital and 1-year mortality in the AMI and COPD inpatients. Variables in the baseline model included age, sex, race (aborigines vs. non-aborigines), and whether the patient received surgery or not. The aboriginality is an important risk factor for health outcomes ,26. We uThe statistical performance of each model was assessed using logistic regression . Three c2 statistics, which resulted from comparing the maximized likelihood function of a model including the comorbidity variables with the nested baseline model. The model comparison statistic has a Chi-squared distribution with the degree of freedom (df) equal to the difference in number of parameters [To assess whether adding the comorbidity measure improved the fit of the regression models, we reported Grameters .To evaluate model discrimination, we reported c statistics, which represents the area under the receiver-operating characteristic (ROC) curve. The c-statistic values range from 0.5 (no greater predictive power than chance) to 1.0 (perfect prediction). Finally, bootstrapping with 1000 replications was conducted to obtain the 95% confidence interval of the c-statistics. This percentile-based confidence interval evaluates the variability of the c-statistic . All anaWe studied 8,961 AMI and 32,755 COPD patients. The proportion of the males and the aborigines in each study population were similar. The average age of patients was 66.31 in the AMI group, and 72.54 in the COPD group. Surgery rates were 14.5% in the AMI patients, and 1.17% in the COPD patients. In-hospital mortality rates were 14.94% in the AMI group, and 3.02% in the COPD group. One-year mortality was 27.07% in AMI patients, and 22.48% in COPD patients in Table First, we compared each logistic model of three methods to the nested baseline model. Table Second, we compared three comorbidity methods when applied to the same outcome, population, and data period. Table Third, we compared model discrimination across data periods within the same outcome, population, and comorbidity measure. Since the Charlson/Deyo and the Charlson/Romano models need information from prior hospitalizations, they performed better when using information from the index and prior hospitalizations compared with the index hospitalization only. In predicting 1-year mortality in the AMI patients, the Charlson/Deyo method had higher c-statistic when using information from the index and prior hospitalizations (0.766) than the index hospitalization only (0.747).However, the Elixhauser models showed different patterns between different outcomes. In predicting in-hospital mortality, the Elixhauser models performed better when using the index hospitalization only. More importantly, in predicting in-hospital mortality, Table Nevertheless, in predicting 1-year mortality, the Elixhauser models using information from the index and prior hospitalizations performed better than using information from the index hospitalization only.To our knowledge, this is the first study that validates the Elixhauser method and compares it to other methods in the Asia region. Further, it is the only investigation that examines the modeling performance of both in-hospital and 1-year mortality of the Elixhauser method, and is the first to report its differences in data periods between short- and long-term outcomes.Several implications for risk adjustment can be drawn from this study. First, this study showed further evidence of external validity of the Elixhauser method in a different dataset and population. In every comparison, it was superior to the Deyo et al. version of the Charlson comorbidity index, which has been used widely for outcome and epidemiology studies. It also outperformed the Romano et al. adaptation of the Charlson index, which has been reported to be better than the Charlson/Deyo method in several studies ,15-17. TSecond, our findings expand upon the results of two earlier studies which used the same analytical method in creating comorbidity variables,7, and dThird, new findings from different data periods provide additional insight into the comorbidity measure. Since some diagnoses are included only when they appear in the prior admissions when using the Charlson/Deyo method and the Charlson/Romano method ,30, thesOne limitation of this study is that the administrative data are claimed for reimbursement purposes rather than research purposes and thus varied in data quality . The quaSince comparative performance of different comorbidity measures can only be examined when other factors, such as population, outcome, and data periods, are all the same. So does the analysis of data periods. The present study examined three types of comparisons by using a manageable design which focused on three measures of comorbidities, two data periods, two diagnoses, and two outcomes. Moreover, such design could become strength for its simplicity to be applied to other populations in Asia or other areas of the world. Similar studies of comparative performance are needed and can be applied to different populations, datasets, outcomes, data periods, and other case-mix methods.In the AMI and COPD patients, the comorbidity measurement developed by Elixhauser et al. has good predictive validity, and researchers should consider its use with claims-based studies rather than the customarily used Charlson/Deyo method. In predicting in-hospital mortality, the Elixhauser method has better discrimination using the index hospitalization; while in predicting 1-year mortality, it may perform better using information from the index and prior 1-year hospitalizations.AHRQ: Agency for Healthcare Research and Quality; AMI: Acute myocardial infarction; CCI: Charlson comorbidity index; CCS: Clinical classifications software; COPD: Chronic obstructive pulmonary disease; DRG: Diagnosis-related group; HCUP: Healthcare Cost and Utilization Project; ICD-9-CM: International Classification of Disease, 9th Revision, Clinical Modification; ROC: Receiver operating characteristic.The authors declare that they have no competing interests.YTC contributed to the study design, statistical analysis, interpretation of data, and writing of the manuscript. YYN contributed to the interpretation of data, and revised the manuscript. SCW contributed to the study design, interpretation of data, and revised the manuscript. All authors have read and approved the final version of the manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1472-6963/10/140/prepubAppendix 1. Comparison of the components of the modelsClick here for file"} +{"text": "HIV and syphilis are major public health problems in Morocco. The region of Souss-Massa, south-west of the country, hold more than 24% of HIV seropositive cases registered in Morocco during 2009. The aim of this study is to evaluate the seroprevalence of syphilis among HIV seropositive patients in the region of Souss-Massa, south-west of Morocco.To evaluate the seroprevalence of syphilis and neurosyphilis among HIV seropositive patients, we retrospectively investigated the medical records of HIV-infected patients attending the regional hospital located in the city of Agadir, during the period comprised between 2011 and 2016.The population studied involved 1381 males (49.18%) and 1427 females (50.82%) HIV seropositive patients. Among them, 481 patients were seropositive for syphilis and three cases were diagnosed with neurosyphilis. The sex ratio distribution was 243 male (52.71%) and 218 female (47.29%). The prevalence of syphilis among the studied population was estimated to 16.42% with a slight dominance in male (17.63%) compared to female (15.28%). By contrast, neurosyphilis was only detected in male patients, with a prevalence estimated to 0.11%.Even if the prevalence of HIV and syphilis is stable in the region of Souss-Massa, the prevalence of syphilis among HIV seropositive patients remained high and correlated positively with that of HIV infection. We did not find a significant difference between the genders, in relation to the prevalence of HIV and syphilis. We concluded that it was essential to continue monitoring the population, in order to improve the prevention and the access to the medical care in the south-west of Morocco. Treponema pallidum [Syphilis is a sexually transmitted infection (STI), associated with the bacterium pallidum . The vaspallidum . Howeverpallidum . During pallidum -6. This pallidum , 8, contpallidum , 10. Thepallidum . Each stpallidum . The ovepallidum -15, partpallidum ; in partpallidum -19. The pallidum , 21. Intpallidum . In 2009pallidum . This leCollecting data: the department of infectious disease of the regional hospital in Agadir covers all HIV seropositive patients from the whole region of Souss-Massa, in Morocco. The medical records of HIV-infected patients, tested between 2011 and 2016, were examined for the presence of cases of syphilis and neurosyphilis.Screening for HIV: screening for HIV infection was performed according to the Moroccan Health Ministry recommendations white visual read; qualitative immunoassay for the detection of antibodies to HIV-1 and HIV-2; or the ELISA test . In addition, a confirmation test was performed using a Western blot test (MP Diagnostics (MPD) HIV BLOT 2.2, Japan). The interpretations of the tests were performed in accordance with the recommendations of the World Health Organization (WHO), based on the detection of two ENV bands, with or without GAG or POL bands.ndations . The diaSerological tests for the detection of syphilis: the serological diagnosis of syphilis was based on a series of two types of serological tests. The first test was a non-treponemal antigen test (VDRL), used for the screening for syphilis in serum or cerebral spinal fluid. The Venereal Disease Research Laboratory test allowed the detection of antibodies directed against non-treponemic antigens, called cardiolipins. The second test was a treponemal antigen test TPHA . This test was based on an indirect hemagglutination assay for the detection and titration of antibodies against the causative agent of syphilis, Treponema pallidum. The samples that were positive in both tests were then registered as seropositive for syphilis.Statistical analysis: the statistical analysis of the data was performed using the R software, version 3.16. The results were summarized using descriptive statistics. The Welch two sample t-test was used to evaluate the differences between gender for both HIV and syphilis prevalence. The Pearson's correlation coefficient was used to assess the correlation between HIV infection and syphilis.Ethical considerations: the data were collected in the hospital register, and the information obtained were kept confidential. The study was approved by the Department of Infectious Disease of the Regional Hospital in Agadir.A total of 2808 HIV seropositive patients were included in the present study. The calculated sex ratio was 0.97, for a gender distribution of 1381 males (49.18%) and 1427 females (50.82%). The average annual incidence of HIV infection between 2011 and 2016 was estimated to be about 468 \u00b1 94.41 cases per year. The highest number of new cases was recorded in 2014, with 623 (22.19%) cases. By contrast, the lowest number of new cases was recorded in 2012 with 346 (12.32%) cases . We did The region of Souss Massa, south-west of Morocco, is home to 2,677 million inhabitants , many of them live in Agadir, the capital and the largest city of this region. The department of infectiology within the regional hospital in Agadir provides medical care and drug to almost every HIV seropositive and STIs patients in the region of Souss-Massa . Like evet al 2016, estimated 496,000 prisoners in MENA, with drug-related offences being a major cause for incarceration [et al. [We noted a stable incidence of new cases of syphilis, despite the peak incidence in 2014. The prevalence of syphilis between 2011 and 2016 was estimated at 17.13% and did not appear to be affected by the genders , consistceration . In Moroceration . But notceration , 32, whiceration -36; especeration . A recenceration . The preceration . The immceration can alsoceration . This wa [et al. . The mal [et al. . The men [et al. . Almost [et al. . Indeed, [et al. . There wBoth HIV and syphilis infections reached alarming rates in the region of Souss-Massa in the south-west of Morocco. Despite the peak recorded in 2014, the prevalence of HIV appears to be stable. However, the prevalence of syphilis among HIV patients remained high, and following the same trend as HIV. In order to prevent or to anticipate any further change in the current situation, it is important to keep a permanent scrutiny of the prevalence and incidence in the region of Souss-Massa. This will be essential to provide a better care and to put in place adapted strategies of prevention in Morocco, especially among the most vulnerable in the general population.Since 2009, high HIV and syphilis prevalence was reported in the region of Souss-Massa, in Morocco;HIV, STIs testing and counseling is a key strategy to reduce sexual risk-taking and control the burden of HIV infection;In the region of Souss-Massa, MSM, FSW and prisoners constituted the main high-risk groups carriers of HIV, syphilis and several sexually transmitted infections.The prevalence of syphilis among HIV-infected patients were stable, over the years but remains very high;There was a significant correlation between the prevalence of HIV and syphilis infections in the Souss-Massa population;Significant efforts will be needed to reduce the prevalence of syphilis and HIV in this region.The authors declare no competing interests."} +{"text": "A health worker dispenses oral cholera vaccine as part of a cholera vaccination campaign in North Kivu in the Democratic Republic of the Congo.A 5-year-old child from the Democratic Republic of the Congo, who entered Uganda with his parents through the Bwera Border post in the country\u2019s Western Region, was confirmed to have Ebola virus disease on 11 June. It was the first confirmed case of Ebola in Uganda, since the Ebola outbreak started in the Democratic Republic of the Congo in August 2018. The case was reported by the Ugandan Ministry of Health and the World Health Organization (WHO). The boy was brought into Kagando hospital in the town of Kasese. Health workers at the hospital identified Ebola as a possible cause of illness and the child was transferred to Bwera Ebola Treatment Unit. The confirmation was made at the Uganda Virus Institute.The Ministry of Health and WHO dispatched a Rapid Response Team to Kasese to identify other people who may be at risk of infection, and to ensure that they are monitored and treated if necessary.https://afro.who.int/news/confirmation-case-ebola-virus-disease-ugandaMore than 730 people have been diagnosed with human immunodeficiency virus (HIV) infections in Larkana, Sindh province, southeast Pakistan. Almost 600 of those diagnosed are children. Before the outbreak, some 1200 children were known to be living with HIV throughout Pakistan.The outbreak was first reported on 25 April 2019. An HIV screening programme was initiated on 28 April, and was expanded on 8 May, with additional health workers being deployed. On 16 May, local authorities established a new antiretroviral treatment clinic for children in Larkana. Early last month they were working to ensure supplies of antiretroviral drugs through global procurement processes, as they continued to screen patients and map the disease outbreak.A team of experts from WHO Headquarters and the Regional Office for the Eastern Mediterranean arrived in Pakistan on 28 May, at the request of the Ministry of Health, to support the response to the outbreak. WHO is coordinating the efforts of an international team, working with local officials.http://www.emro.who.int/pak/pakistan-news/who-supports-response-to-hiv-outbreak-in-sindh-pakistan.htmlA cholera vaccination campaign launched in North Kivu in the eastern part of the Democratic Republic of the Congo on May 27, reached 762 159 of the targeted 800 000 people within the first week. The first of two doses of oral cholera vaccine was administered. At the beginning of June there were tentative plans to conduct the second dose campaign in North Kivu between 9 and 13 July.The campaign is being implemented by the Ministry of Health with support from WHO and partners and is funded by Gavi, the Vaccine Alliance.More than 10 000 cases of cholera have been reported in the country since January 2019, leading to more than 240 deaths.https://afro.who.int/news/major-cholera-vaccination-campaign-begins-north-kivu-democratic-republic-congoTransmission of Ebola virus disease cases in the outbreak in the Democratic Republic of Congo appeared to be easing at the beginning of June, with confirmed cases dropping to around 90 per week at the beginning of the month, down from a peak of 126 cases per week reported in April.Declines in the incidence of new cases have been most noticeable in hotspots such as Katwa, Mandima and Beni health zones. The outbreak continues to be contained within 12 active health zones in North Kivu and Ituri provinces.One possible reason for the decline was an improved security situation, which was allowing response teams to implement key interventions including infection prevention and safe burial practices.As of 6 June 2019, a total of 2025 cases had been reported and 1357 deaths, representing an overall case fatality ratio of 67%. Among those infected were 110 health-care workers, representing 5% of total cases.https://www.who.int/csr/don/06-june-2019-ebola-drc/en/New WHO estimates highlight the need for increased, sustained investment in the development of mental health services in areas affected by conflict. Lancet on 12 June, roughly one person in five (22%) is living with some form of mental disorder, ranging from mild depression or anxiety to psychosis.According to the estimates published in the Overall, the mean prevalence was highest for mild mental health conditions (13%), for moderate conditions the prevalence was 4%, and for severe conditions the prevalence was 5%. Depression and anxiety appeared to increase with age in conflict settings and depression was more common among women than men.The findings suggest that past studies underestimated the burden of mental health conditions in conflict-affected areas, with higher rates of severe mental illness and also of mild to moderate mental health conditions.http://www.thelancet.com/journals/lancet/article/PIIS0140-6736(19)30934-1/fulltextEpilepsy: a public health imperative was produced by WHO and key partners and published on 20 June. The first global report on epilepsy, entitled, The report highlights available evidence on the burden of epilepsy and the public health response required to address it, and calls for increased action on epilepsy, including the addressing of gaps in research. Most people with epilepsy live in low- and middle-income countries and do not have access to treatment. However, effective antiseizure medicines can cost as little as US$ 5 per year and evidence shows that epilepsy interventions can be successfully integrated into primary health care in many countries.https://www.who.int/mental_health/neurology/epilepsy/report_2019/enWHO launched its first consolidated guideline on self-care interventions for sexual and reproductive health on 24 June. WHO consolidated guidelines on self-care interventions for health: sexual and reproductive health and rights, the guidelines make evidence-based recommendations on a range of interventions including self-administration of injectable contraception, use of home-based ovulation predictor kits for fertility management, and self-collection of samples for sexually transmitted infection testing.Entitled The guidelines are the first of a planned series, which are expected to cover other health topics as new evidence emerges.https://www.who.int/reproductivehealth/self-are-interventions/en/A woman returns from her garden near the Mirami point of entry between Uganda and the Democratic Republic of the Congo, where Uganda Red Cross Society supported by the United Nations Children\u2019s Fund and other international agencies, including WHO, are leading an Ebola screening intervention, as part of the response to the ongoing Ebola virus outbreak.A clinical study supported by WHO and partners and conducted in four African countries found no significant difference in the risk of HIV infection among women using one of three contraceptive methods.Lancet on 13 June, the Evidence for Contraceptive Options and HIV Outcomes study was done to address concerns raised by evidence from observational studies\u00a0that use of progestogen-only injectable methods, particularly\u00a0depo-medroxyprogesterone acetate, might be associated with an increased risk of acquiring HIV.Published in the The study compared three highly effective, reversible methods of contraception to evaluate whether there was any difference in the risk of acquiring HIV infection among users of these methods.http://echo-consortium.com/the-evidence-for-contraceptive-options-and-hiv-outcomes-echo-study-2/More than 23 million individuals in the WHO European Region become sick from eating contaminated food every year, some 4700 of them dying as a result. Diarrhoeal diseases are responsible for 94% of food-related illnesses, and 63% of food-related deaths.The burden of foodborne diseases in the WHO European Region and were presented as part of the first-ever World Food Safety Day on 7 June.The data were published in a report entitled An estimated 600 million people, almost 1 in 10 people in the world, fall ill after eating contaminated food and 420 000 die every year.https://www.who.int/foodsafety/publications/foodborne_disease/fergreport/en/RESPECT women: preventing violence against women, was launched on 29 May and is aimed primarily at policy-makers.WHO, in collaboration with UN Women and 11 partners, launched a new framework for interventions and programmes designed to prevent violence against women. The initiative, called About 1 in 3 women worldwide has experienced either physical and/or sexual violence in their lifetime, according to WHO estimates.https://www.who.int/reproductivehealth/topics/violence/respect-women-framework/en/23 September - United Nations High-Level Meeting on universal health coverage. UN Headquarters, New York, United States of America.24\u201325 September - Sustainable Development Goals Summit, New York, United States of America.22\u201324 October - Fifth High-level Meeting on Transport, Health and Environment, Vienna, Austria."} +{"text": "Nature Communications 10.1038/s41467-018-03247-3; published online 26 February 2018Correction to: In the original version of this Article, the affiliation details for Qiushuo Shen incorrectly omitted \u2018Kunming College of Life Science, University of Chinese Academy of Sciences, Kunming, 650204, China\u2019. This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "In the original article, we neglected to include the acknowledgment of the TERRA REF project, funded by the Advanced Research Projects Agency-Energy (ARPA-E), U.S. Department of Energy, under award number DE-AR0000594.The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated."} +{"text": "The European Centre for Disease Prevention and Control (ECDC) invites applications for the 2019 ECDC traineeship programme including the following areas.Eurosurveillance editorial office: the office is responsible for Eurosurveillance; Europe's journal on infectious disease surveillance, epidemiology, prevention and control published by ECDC.Vaccine-preventable diseases: the programme is responsible for epidemic intelligence, health communication, microbiology, preparedness and response, public health training, scientific advice and surveillance for vaccine-preventable diseases.Influenza and other respiratory viruses: the programme is responsible for epidemic intelligence, health communication, microbiology, preparedness and response, public health training, scientific advice and surveillance for influenza and other respiratory viruses.Stakeholder engagement: the function is responsible for improving the communication with ECDC's stakeholders. The tasks of the trainee will be shared between the communication and the public health training section.Country capacity support and capacity gap analysis: the section is responsible for coordinating ECDC's activities in planning and evaluation of emergency preparedness and other systems for communicable disease control.https://ecdc.europa.eu/en/about-uswork-us/ecdc-traineeship-programmeFor more information, please visit the ECDC website:"} +{"text": "There are errors in the Funding section. The correct funding information is as follows: This study was funded by the Universiti Kebangsaan Malaysia grant DPP-2018-134.The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript."} +{"text": "This study aimed to examine factors associated with life-space constriction, using the data from the Health and Retirement Study, a nationally representative sample. We limited our analysis to those who were 65 years and older and answered to the 2012 experimental module on life-space . Life-space was assessed with the modified version of the UAB Study of Aging Life-Space Assessment, ranging nine zones: room, home, own property, immediate neighborhood, town, community, county, state, and region. A series of logistic regression models were used to estimate odds ratios for life-space constriction by sociodemographic and health characteristics. The results showed that 3.0% and 6.7% of older adults reported that they had never been to places beyond their home and own property/apartment building for the past four weeks, i.e. the critical boundaries in terms of social isolation. The significant factor associated with the life-space constriction within home, immediate neighborhood, and town was physical mobility limitation , while the constriction within county was associated with education level (OR: 0.91). Driving a car was negatively associated with the life-space constriction within own property/apartment building and home . Policy makers need to pay more attention to social and environmental factors influencing social isolation among older adults such as transportation options and social class disparity."} +{"text": "Communications Biology; 10.1038/s42003-018-0098-3; published online 17 July 2018Correction to: In the original HTML version of the paper, the following affiliation was missing for author Cheong Xin Chan: School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD 4072, Australia. This affiliation was incorrectly assigned to author Pim Bongaerts as \u201cPresent Address\u201d. All affiliations were published correctly in the PDF version of the paper and have now been corrected in the HTML."} +{"text": "Escherichia coli, Bacillus subtilis, and Saccharomyces cerevisiae). Machine-learning models were constructed to predict a selection of synonymous codons (low- or high-frequency-usage codon) in a gene. This method could be easily and efficiently used to design new genes from protein sequences for optimal expression in three expression hosts . Presyncodon is free to academic and noncommercial users; accessible at http://www.mobioinfor.cn/presyncodon_www/index.html.In the natural host, most of the synonymous codons of a gene have been evolutionarily selected and related to protein expression and function. However, for the design of a new gene, most of the existing codon optimization tools select the high-frequency-usage codons and neglect the contribution of the low-frequency-usage codons (rare codons) to the expression of the target gene in the host. In this study, we developed the method Presyncodon, available in a web version, to predict the gene code from a protein sequence, using built-in evolutionary information on a specific expression host. The synonymous codon-usage pattern of a peptide was studied from three genomic datasets ( In most organisms, 61 universal genetic codons encode for 20 standard amino acids, of which 18 are encoded by multiple synonymous codons. In all domains of life, a biased frequency of synonymous codons is observed at the genome level. Many studies have proved that the presence of synonymous codons in the gene coding regions is not inconsequential, and relates to the efficient and accurate translation of the protein . TherefoMany methods, including JCat , Gene DeEscherichia coli, Bacillus subtilis, and Saccharomyces cerevisiae). This big data gene prediction method was used to learn the codon-usage pattern for a peptide as-derived from sequenced genomic data. Machine-learning models were constructed by the random forest classification to predict a selection of synonymous codons (low- or high-frequency-usage codon) for the target gene. Compared with the early version of Pyesyncodon, which could only design the gene to be efficiently expressed in E. coli in local [E. coli, B. subtilis, and S. cerevisiae) on the web; and the training dataset has been updated with more genomes. Therefore, this method will be easily and efficiently used to design genes for heterologous gene expression in the three popular expression hosts .To address the need for heterologous gene design, based on all used codons (the high- or low-frequency-usage codons), a new web server application, Presyncodon, was developed to design the heterologous gene for expression in the three frequently-used recombinant hosts were constructed, which contained 353, 62, and 20 genomes, respectively. All selected genomes were the complete genomes downloaded from NCBI (ftp://ftp.ncbi.nlm.nih.gov/genomes/) on July 13th, 2016, and their genome accession numbers are shown in Three genomic datasets were also constructed. The software CD-HIT [B. subtilis, E. coli and S. cerevisiae, were constructed with 8091, 11232, and 5905 genes from the total 1461067, 256246, and 107820 genes, respectively. In order to train the predicting models, three non-redundant gene datasets , expected maximal score (m) of the target fragment, and the matched percent against the CSI file were calculated by the method described in [c), if the calculated matched percent of multiple fragments from the CSI file for a fragment was greater than the cut-off level, the coding vector for the middle codon in the fragment was the arithmetic average of those vectors encoding the selected multiple fragments. Here, the training label is the codon for the middle amino acid.The training gene sequences were translated, and were also split into window sizes of five or seven amino acids, and searched against the corresponding CSI files. For each fragment, the matched score ,23 was u4) in case of the five residues\u2019 long fragments and of 1152000000 (18 \u00d7 205) in case of the seven residues long fragments) were searched against the corresponding CSI files, with four cut-off levels . Input vectors were generated for each fragment and the synonymous codons selection, based on the distribution of the middle residue in the fragment, was predicted for each organism. The results were stored in the PostgreSQL database.In order to increase the speed of target gene design, all possible fragments , were evaluated by ten-fold cross validation. As shown in c) achieved higher accuracy than those obtained with smaller cut-off levels. Therefore, the codon-usage tendency for each amino acid could be predicted by only one model, as obtained from the long-window-sized amino acid fragments and characterized by a larger cut-off level (c). The first and last two codons were selected statistically and cut-off level . In the next step, we will develop this optimizing method for more expression systems. Therefore, this method could be easily used to design synthetic genes for heterologous gene expression in biotechnology.The software Presyncodon is designed as an adaptable, web-based interface that could be easily used by scientists. This website was built using Linux (Centos ver. 6.5), Apache (ver. 2.2), PostgreSQL ver. 8.4.20), and Perl (ver. 5.10.1). The input of the user is the target protein sequence and the only external parameter required is the selection of the target expression host . The wai, and Per"} +{"text": "Background: Rheumatoid Arthritis (RA) is a systemic autoimmune disease leading to joint destruction. The prevention of bone and cartilage destruction has received increased attention in recent years.Objective: To evaluate the current evidences regarding the bone-protecting efficacy of Chinese medicine or the combination of Chinese medicine and Western medicine for RA.Methods: We comprehensively searched PubMed, Embase, the Cochrane Library (www.thecochranelibrary.com), the China National Knowledge Infrastructure (CNKI), the Database for Chinese Technical Periodicals (VIP), and SinoMed. We then performed a systematic review and cumulative meta-analysis of all randomized controlled trials (RCTs) assessing the two therapy methods.Results: Sixteen studies including 1,171 patients were included in the final analysis. The results showed that Chinese medicine could significantly improve the bone mineral density (BMD) , and decrease the serum matrix metalloproteinase 3 (MMP-3) .Conclusions: Chinese medicine may provide an efficiently alternative choice for the treatment of RA in terms of the bone-protecting efficiency. Given the inherent limitations of the included studies, future well-designed RCTs are required to confirm and update the findings of this analysis. Rheumatoid Arthritis (RA) is a chronic systemic autoimmune disease with symmetric inflammation of aggressive multiple joints Miossec, . As the Up to date, there are no systematic reviews and meta-analysis regarding bone-protecting efficiency about Chinese Medicine alone or in combination with Western Medicine in the treatment of RA. We therefore systematically searched and analyzed the available literature to evaluate the efficacy and potential advantages of Chinese Medicine (or a combination of Chinese and Western medicine), when compared with Western Medicine.All randomized controlled clinical trials.Adults with a diagnosis of RA either using the 1987 American College of Rheumatology (ACR) classification criteria and the levels of serum matrix metalloproteinase 3 (MMP-3), a biomarker for bone destruction , inception to 31 December 2017;The Cochrane Library , inception to 31 December 2017;The Database for Chinese Technical Periodicals (VIP), inception to 31 December 2017;SinoMed, inception to 31 December 2017.In addition, we manually searched the reference lists of the included studies and previous review papers to find additional studies. All references were imported to an EndNote (x6) library and tagged with the name of the database.Two review authors assessed the titles and abstracts for all the records identified through the search strategies, retrieving full texts for all those that appeared to satisfy the following criteria: the type of study; type of participants; type of intervention; type of measurements. Data from the included studies were extracted and summarized independently by at least two of the authors. Any disagreement was resolved by a discussion among all the authors.For data extraction, the review team allocated papers to different authors according to their areas of expertise, and two reviewers independently retrieved the details for each publication and tabulated them in a standardized form. The retrieved details include intervention (including characteristics and duration), assignment to groups , outcome measures, timing of measurements, adherence to intervention/control, sample size statistical analysis methods as well as adverse events and withdrawals. Two review authors independently extracted data from the reviews using a predefined data extraction form created as a Microsoft Excel\u00ae spreadsheet.Studies were rated for evidence level according to the criteria given by the Centre for Evidence-Based Medicines in Oxford, UK , standardized mean differences (SMD), and 95% confidence intervals (CIs) were calculated for the continuous outcomes (reporting mean and standard deviation of the mean). Where the standard deviations were not explicitly stated, we calculated them from the different means and their respective CIs or I2 statistics , a random-effects model would be used to further analyze the results. Where we detected this level of heterogeneity and there were sufficient studies available, we conducted subgroup analyses in an attempt to explain the heterogeneity.Where appropriate, we formally assessed heterogeneity of the data using the We used Egger's test to assess the possibility of publication bias with Stata 11.0.Where there was no heterogeneity, we used a fixed-effect model, and where there was heterogeneity, we used a random-effects model if there was no clinical heterogeneity. The MD for pooled data in meta-analysis were calculated using a fixed model as outcomes were measured on the same standard scales. Otherwise, the SMD were calculated. Meta-analysis was facilitated by Review Manager 5.3 using the statistics as described below.Where sufficient studies were available and the data was heterogeneous, we carried out separate meta-analyses for studies according to some factors including intervention duration, disease stage, and the form of traditional Chinese medicine (TCM).In studies where calcium supplements were used, we planned the sensitivity analyses as a priority in order to explore the differences in effect size and to assess whether the conclusions were robust to the decision-making process. The sensitivity analyses included the following:(1) The effect of risk of bias in included studies\u2014defined as adequate allocation concealment and blinding of outcome assessors; (2) the effect of using calcium supplements.We undertook a comprehensive literature search, including screening of titles and abstracts. In total, we retrieved 25 fulltext references for further evaluation, including the manual searching of reference lists (9 further full-text studies).Sixteen studies were excluded: two did not explain the diagnostic criteria, six did not include the outcomes of interest, and one used proprietary Chinese medicine in the control group.Overall, for most of the included studies the risk of bias was low or unclear. The methodological quality summary for each included study is presented in Figure Investigators described all the studies included as randomized controlled trials. Nine studies Figure .I-squared was 98% and the p-value was <0.00001, so a random effects model was adopted for the meta-analysis. Significant differences were found in the reduction of serum MMP-3 levels between the Chinese medicine (or Chinese combined with Western medicine) group and Western medicine group Figure .P = 0.42).Two studies Liu, includinP = 0.03).Five studies .Data from one paper .Two studies Figure .P = 0.11).Two studies Su, includinP = 0.13) Figure .P < 0.0001).Six studies . There is significant publication bias for MMP-3 (P = 0.011) (Table Egger's test of BMD did not suggest significant publication bias (1) Table .The aim of this review was to provide an overview of bone-protecting effects of Chinese medicines in the treatment of RA. The review revealed 16 RCTs investigating bone-protecting interventions.The changes in local metabolism may impair the dynamic balance of bone formation and resorption, finally leading to bone and cartilage destruction of RA compared with Western Medicine. In assessing the extent of bone destruction, imaging provides us with a more intuitive evidence. The pooled data of outcomes indicates that Chinese Medicine can improve the imaging findings of RA. Some studies is more effective in improving the BMD than only Western medicine treatment.Pooled data indicated that the MMP-3 level was reduced in the Chinese medicine group. It is known that the MMPs can degrade all the protein components of the cartilage, resulting in destruction of ligaments, cartilage, and bone had a better effect in comparison to only Western medicine in reducing the destruction of cartilage.This meta-analysis of 16 RCTs or CCTs includes 1,171 patients. Our findings suggest that the Chinese traditional medicine leads to a statistically significant increase in the BMD and decrease in MMP-3, which implies that the Chinese medicine may provide an efficient treatment option for RA in terms of the bone-protecting efficiency, especially to patients in China. However, the limitations of this meta-analysis are also to be noted. All the included studies were performed only in China. Many of the studies included a small number of patients. The most important criteria, including the BMD, imaging stage, and MMP-3, were not reported in all the studies. In addition, the limited numbers of RCTs prevented us from reaching any definitive conclusions. Furthermore, bone destruction is a chronic progression, but the study of the literature for 3\u20136 months does not give long-term results. Therefore, more studies with high quality are urgently needed to judge the full potential of the Chinese traditional medicine for use in bone-protection. Accordingly, the conclusions of this review should be carefully interpreted. Due to the increasing use of Chinese medicine, accurate and complete data on the interactions between Chinese medicine and western medicine are urgently required.XC, X-MC, R-YH, Z-HW, and P-JJ contributed to the literature database search, data collection, data extraction, data analysis, and writing of the manuscript. XX, KB, R-RW, J-HP, H-JL, Q-WY, J-YY, M-JW, HY, J-JL, Y-JH, and Q-CH performed data analysis and rationalization of the results. The topic was conceptualized by P-JJ, R-YH, and Z-HW.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Scientific Reports 10.1038/s41598-019-47796-z, published online 22 August 2019Correction to: The original version of this Article contained an error in the spelling of the name of the author Amy Silder, which was incorrectly given as Amy B. Silder. This has now been corrected in the PDF and HTML versions of the Article, and in the accompanying Supplementary Material file."} +{"text": "Correction to: Environmental Healthhttps://doi.org/10.1186/s12940-019-0454-xFollowing publication of the original article , the aut\u2018Competing interests\u2019 in the original article.The authors declare that they have no competing interests. All willingly contributed their knowledge and perspective on the conduct and interpretation of two toxicology studies for chlorpyrifos and chlorpyrifosmethyl, respectively.Revised \u2018Competing interests\u2019.At the time of submission, Drs. Juberg and Marty were employed by Dow AgroSciences LLC and The Dow Chemical Company, respectively. Dow AgroSciences is the primary registrant of chlorpyrifos and chlorpyrifos-methyl in the U.S., and The Dow Chemical Company is a manufacturer of both chemicals. All other co-authors are not affiliated with either company. All co-authors willingly contributed their knowledge and perspective on the conduct and interpretation of two toxicology studies for chlorpyrifos and chlorpyrifos-methyl."} +{"text": "Microcontrollers are programmable, integrated circuit chips. In the last two decades, their applications to industrial instruments, vehicles, and household appliances have reached the extent that microcontrollers are now the number-one selling electronic chip of all kinds. Simultaneously, the field of lab-on-a-chip research and technology has seen major technological leaps towards sample handling, sample preparation, and sensing for use in molecular diagnostic devices. Yet, the transformation from a laboratory based lab-on-a-chip technology to actual point-of-care device products has largely been limited to a fraction of the foreseen potential. We believe that increased knowledge of the vast possibilities that becomes available with open source microcontrollers, especially when embedded in easy-to-use development environments, such as the Arduino or Raspberry Pi, could potentially solve and even bridge the gap between lab-on-a-chip technology and real-life point of care applications. The profuse availability and extraordinary capabilities of microcontrollers, namely within computation, communication, and networking, combined with easy-to-use development environments, as well as a very active and fast moving community of makers, who are eager to share their knowledge, could potentially be the difference between a dreadful \u201cchip-in-a-lab\u201d-situation, and the next successful start-up. Here follows a brief insight into how open source microcontrollers could potentially have a transformative effect on the field of lab-on-a-chip research and technology. Details in some specific areas of application are briefly treated before addressing challenges and future perspectives. Lab on a chip (LoC) technology has been a part of the analytical chemistry community since the early 90s ,2. The gBrief CV of the authorsDang Duong BangAnders WolffTrieu NguyenSune Z. AndreasenProf. Dang Duong Bang received his PhD in 1995 on Molecular biology from Leiden University, Leiden, and the Netherlands. In 2002, he was appointed as senior researcher and leaded the Laboratory of Applied Micro and Nanotechnology (LAMINATE) at National Veterinary Institute, Technical University of Denmark. In 2014, he was promoted as Professor MSO at National Food Institute, Technical University of Denmark. His research focuses on development of total integrated Lab on a chip systems for clinical diagnosis of infectious and food borne diseases. Professor Dang contributed more than 130 papers published in international peer review scientific journals and owned 5 patents.Assoc. Prof. Anders Wolff received his M.Sc. in Chemical Engineering from Technical University of Denmark (DTU) in 1993 and his PhD in Biochemical and Engineering from Delft University of Technology, the Netherlands in Dec 1997. In 1998, Dr. Wolff joined the Department of Micro and Nanotechnology . In 2000, he was appointed the associated professor and leaded the Cell Handling Group (now BioLabChip) at DTU. His research interests: PCR chip with integrated heaters and thermos sensor, integrated microsystem for sample preparation and DNA amplification. Dr. Wolff owned 7 patents, 80 papers published in international scientific journals.Dr. Trieu Nguyen obtained his PhD (2015) on the microfluidic energy conversion with prof. Jan Eijkel and prof. Albert van den Berg . From November 2015 to January 2017, he worked as a postdoc on microfluidic mixers for protein folding studies in Michigan State University, USA. Currently he is a postdoc researcher at National Food Institute, Technical University of Denmark. His research interests stay on micro-, nanofluidics, Lab on a chip, microfabrication, physical chemistry, electrochemistry and rapid detection of food-borne diseases.Dr. Sune Z. Andreasen received his M.Sc. in physics and nanotechnology from Technical University of Denmark (DTU) in 2013, and a PhD from the same place in 2017 on Lab-on-a-chip devices, specialized on centrifugal microfluidics, electrochemistry, and automated sample preparation. Since then, he has worked as a postdoc at DTU Nanotech developing sensing platforms for biochemical reactions, with emphasis on commercially viable solutions. His research interests include LoC applications, microfluidics, open-source electronics, instrument development and entrepreneurship.As mentioned earlier, microcontrollers are low-cost, single chip computers . The hisWe believe that recently scientists in general, and particularly in the field of LoC, have started to notice the raising and beneficial role of open-source microcontroller platforms. For instance, a search using the keyword \u201cmicrocontroller\u201d in the Lab on a Chip journal gave the results of 31 articles published recently . Another search using the keywords \u201cmicrocontroller\u201d plus \u201cpoint of care\u201d on Web of Science gave approximately the same result: 32 articles published between 2000 and 2018. A search with the keywords \u201c((lab-on-a-chip OR microfluidics OR point-of-care) AND (microcontroller OR arduino OR teensy OR \u201craspberry pi\u201d))\u201d in Web of Science gave a result of 73 articles, most of them published recently, from 2010 to 2017, and with an increase in number of articles within the recent year shown in . This reThis feature review aims to rise, and hopefully speed up, the awareness in the LoC community, on the beneficial and important role of open source hardware, and especially of microcontroller-based platforms, as we believe we are now at the frontier of the future of PoC devices. Adding to this point, Nature journal reported that there was a conference held in 2016 at CERN, Europe\u2019s particle-physics laboratory near Geneva, Switzerland, with exactly the intention and hope to rapidly increase the awareness about open science hardware solutions to the research community .In the field of LoC research, especially when aspiring towards PoC systems, the type of signals that are readable (or otherwise detectable) to the end users is to a large extent based on (but not limited to) capturing of image(s) or ohttp://openpcr.org/.The third example is the very ambitious \u201cFlyPi\u201d\u2014a \u201c100\u20ac Lab\u201d by A.M. Chagas et al. (2017) . Here, aThe last example is the Proportional-Integrate-Derivative (PID), controlled syringe pump, developed by J.R. Lake et al. . Here, aIn order to help beginners to easily get started on building a prototype from scratch, we here outline some guidance. https://www.arduino.cc/en/Main/Products for an overview of all Arduino products and their specifications. Among the Arduino family of boards, the Arduino Uno is the most popular one, and with good reason, as it is both cheap and reliable. The Uno features fourteen digital input/output pins and six analog inputs. Most of the work done in the do-it-yourself community (that requires the use of a microcontroller) has been conducted on the Uno. Using an Arduino Uno hence offers the users a great access to help and support from the community such as the Arduino Forum. If more than fourteen digital I/O are needed, a Mega Arduino board can advantageously be used, as this development board provides the user with fifty-four I/O digital pins. Apart from the development boards, the Arduino platform also features the open-source Arduino IDE (Integrated Development Environment) software, making writing and uploading of code to the boards an easy and simple task. The IDE can be downloaded at the Arduino website https://www.arduino.cc/en/Main/Software#download. In the below section we give some instructions to, as well as provide some references for, controlling of temperature, pressure and movement, using an Arduino Uno.The Arduino family of open source microcontroller-based boards is a good starting point for beginners to build their own devices. The Arduino family of development boards include Uno, Nano, Micro, Mega, Mini, Leonardo, etc. See http://openpcr.org where they built up a PCR thermocycler based on an Arduino Uno. The source code and full list of components are available at their website, and it is possible to buy a completely open source PCR for 499 USD at their website. For the PID library used to control temperature with an Arduino, the most popular one seems to be the one developed by Brett Beauregard, and the software library is available for download from http://brettbeauregard.com/blog/2011/04/improving-the-beginners-pid-introduction/ or https://github.com/br3ttb/Arduino-PID-Library/blob/master/PID_v1.h.For temperature controlling and measurement, the Arduino Uno is widely used, for instance in references ,30,31,32For creating and controlling of precise movements, an Arduino Uno can also be used to control one or more stepper motors. A stepper motor is a special type of motor that moves in discrete steps. A stepper motor is therefore a good choice for a project requiring very precise movements, such as a 3-D printer (ILIOS HD kit for instance ), or a mFurthermore, stepper motors controlled by an Arduino Uno can also be used to generate and control pressure in a microfluidic system, as shown in example 4 in the previous section. IDE code and bill of materials can be downloaded from the website provided in reference . AlternaThe first challenge is that the beginners who try to build a device based upon an open source microcontroller may feel the task overwhelming. Nevertheless, a little effort invested to learn the basics of how to handle an open source microcontroller, such as an Arduino board, will quickly pay off many times over, as beginners start to build up their own devices and projects. Another challenge is that equipment and devices based on other people\u2019s open source hardware solutions, often are still at an early stage of a product\u2019s evolutionary process. Commercially bought devices hence may provide longer product life times and more robust calibrations/functionality. Nonetheless, as more ideas and designs are contributed, the degree of complexity of open-source devices will grow fast. Eventually, not only the scientific community but also the public will benefit due to an increased availability of low-cost, yet highly sophisticated, open source devices.https://www.smb.dk/smb/quickvet), some are still in the academic research stage . In each stage, the challenges are obviously different, yet we have no doubt that the opportunities of achieving success on those PoC devices are now within reach for much larger parts of the LoC research field, due to the rise of open source communities, especially the ones based on open source hardware, such as the Atmel microcontrollers of the Arduino boards. As Isaac Newton once wrote: \u201cIf I have seen further it is by standing on the shoulder of Giants.\u201d [Some PoC devices have been fully commercialized . In the in 1676) . Togethein 1676) ."} +{"text": "Cell & Bioscience.Two research papers, one from a group led by Dr. Haifan Lin of Yale University School of Medicine, New Haven, USA and another from two laboratories led by Drs. Yihong Ye of National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, USA, and Ting Zhang of Southern University of Science and Technology, Shenzhen, China, respectively, won the 2017 Ming K. Jeang Award for Excellence in Cell & Bioscience in 2018 have been selected to receive the Ming K. Jeang Award for Excellence in Cell & Bioscience. The Ming K. Jeang Award for Excellence in Cell & Bioscience was established in 2011 with a generous donation from the Ming K. Jeang Foundation to honor outstanding research articles published in Cell & Bioscience, the official journal of the Society of Chinese Bioscientists in America . A committee of Cell & Bioscience Editors, chaired by Dr. Dong-Yan Jin, considered all research articles published in the journal in 2018 to select the following two articles to receive the award [We are very pleased to announce that two research articles published in Winifred Mak, Jing Xia, Ee-Chun Cheng, Katie Lowther and Haifan LinCell & Bioscience 2018 8:54.Yilin Ye, Suk-Hwan Baek, Yihong Ye and Ting ZhangCell & Bioscience 2018 8:46.Congratulations to these two groups of investigators for jobs well done!We are looking forward to receiving contributions of outstanding research articles from the scientific community in 2019 and beyond."} +{"text": "Nature Communications; 10.1038/s41467-018-07610-2; published online 29 Nov 2018.Correction to: The original version of this Article omitted a declaration from the Competing Interests statement, which should have included the following: \u2018J.D.B. is a founder and Director of the following: Neochromosome, Inc., the Center of Excellence for Engineering Biology, and CDI Labs, Inc. and serves on the Scientific Advisory Board of the following: Modern Meadow, Inc., Recombinetics, Inc., and Sample6, Inc.\u2019. This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "Scientific Reports 10.1038/s41598-017-09554-x, published online 31 August 2017Correction to: In the original version of this Article, Affiliation 5 was incomplete and incorrectly listed as \u2018Department of Translational Genomics, University of Cologne, 50931, Cologne, Germany\u2019 The correct affiliation is listed below:Department of Translational Genomics, Medical Faculty, University of Cologne, 50931 Cologne, GermanyIn addition, the Acknowledgements was incomplete. It now reads:\u201cThis work was supported by Center for Molecular Medicine Cologne (CMMC) to RB and MO and by the German Cancer Aid as part of the Interdisciplinary Oncology Centers of Excellence program to the Center for Integrated Oncology K\u00f6ln Bonn. In addition, the project was supported by the Federal German Ministry of Science and Education (BMBF) as part of the e-Med program (grant no. 01ZX1303A and 01ZX1603A to RB and RKT) and the German federal state North Rhine Westphalia (NRW) and by the European Union as part of the PerMed.NRW initiative (grant 005-1111-0025 to RKT and RB) as well as the EFRE initiative , by the German Consortium for Translational Cancer Research (DKTK) Joint Funding program (RKT) and a grant from the Volkswagenstiftung (Lichtenberg program) to MS. We highly appreciate the excellent technical assistance of Michael Gentz, Olivia K\u00e4sgen and Marion M\u00fcller.\u201dFinally, disclosures were omitted from the Competing Financial Interests statement which now reads:R.K.T. is a consultant of NEO New Oncology GmbH and received honoraria from AstraZeneca, Bayer, NEO New Oncology GmbH, Boehringer Ingelheim, Clovis Oncology, Daiichi-Sankyo, Eli Lilly, Johnson & Johnson, Merck KGaA, MSD, Puma, Roche and Sanofi.These errors have now been corrected in the PDF and HTML versions of the Article, and in the accompanying Supplementary Information file."} +{"text": "GBD 2017 Causes of Death Collaborators. Global, regional, and national age-sex-specific mortality for 282 causes of death in 195 countries and territories, 1980\u20132017: a systematic analysis for the Global Burden of Disease Study 2017. Lancet 392: 1736\u201388\u20142018; The bottom row in figure 7 was cut off. This correction has been made to the online version as of Nov 9, 2018, and has been made to the printed Article."} +{"text": "SRL vs.ERL Theory predicts that a student's own self-regulation and the regulatory nature of the context are factors that jointly determine the student's level of motivational-affective variables. However, this principle has not yet been verified in the case of achievement emotions. The aim of this research was to test this prediction, with the hypothesis that students' level of self-regulation (low-medium-high), in interaction with the regulatory nature of the teaching (low-medium-high), would determine positive or negative emotions as well as the degree of burnout/engagement. A total of 440 university students completed validated questionnaires on self-regulation; regulatory teaching; achievement emotions in class, in study and in testing situations; and on burnout/engagement. Using a quasi-experimental design by selection, ANOVAs and MANOVAs were carried out. The results confirmed that the level of self-regulation and the level of external regulation jointly determined university students' level of achievement emotions, as well as their level of burnout/engagement. Based on these results, a five-level progressive scale was configured. We conclude that this scale may be useful and adequate as a heuristic technique or model for understanding and analyzing the type of student-teacher interaction that is taking place in the university classroom, and thereby learn the probability of stressful effects and the students' level of emotional health.The On one hand is intelligence, with its related lines of research, such as the study of cognitive and metacognitive factors in learning processes. On the other hand is personality, as well as students' motivational-affective and meta-motivational processes. Detailed analysis over the past years has produced a considerable amount of research evidence, and a paradigm has emerged for the study of emotions and non-cognitive or \u201csoft\u201d skills in the educational sphere were found to have direct relations with students' self-regulation and moderate the relationships between the learning environment and self-regulation variables has to do with positive proactivity, that is, the individual actively and adequately regulates and manages his or her own conduct. This level is referred to as high level, in terms of the degree to and quantity of behaviors used to regulate one's own behavior (level 3).(1) Non-Regulation (NR) may be conceptually defined as a person's lack of proactivity, or the absence of self-regulating behaviors. This is the conceptual equivalent of reactivity. This level is referred to as medium level, in terms of the degree to and quantity of behaviors used to regulate one's own behavior (level 2).(2) Dysregulation (DR) has to do with negative proactivity, that is, the individual actively but inadequately regulates and manages his or her own conduct. Examples include the use of self-handicapping strategies or procrastination. This level is referred to low level, in terms of the degree to and quantity of behaviors used to regulate one's own behavior (level 1). The three behavior types are shown in (3) External Regulatory (ER) context. Positive or adequate proactivity is promoted through the context, which clearly fosters self-regulation. This context features high levels (level 3) of external signs or encouragements to promote self-regulated behavior and increases its likelihood at each moment of learning acts: beginning, middle and end. Such encouragement can be in the form of antecedents or contextual consequences .(1) External Non-Regulatory (ENR) context. The context neither encourages self-regulation nor does it tend to dysregulate students' learning. Medium level or no external signs (level 2) or other aspects promote self-regulated behavior or dysregulated behavior, so as to make either of these more likely at the beginning, middle and end of learning acts. A non-regulatory context supposes that the individual would engage in a moderate level of self-regulated behavior, in the absence of contextual elements that enhance or discourage such action. The context is characterized by a lack of predictability of action.(2) External Dys-Regulatory (EDR) context, actively promotes dysregulation or low levels of self-regulation (level 1). The context promotes proactivity that is not positive, but inadequate or negative. Many external signs make dysregulated behavior more likely, and encourage active dysregulation at the beginning, middle and end of learning acts. These signs can also be in the form of antecedents or contextual consequences that favor dysregulation. This kind of context would require the individual to make a great effort if self-regulation is pursued. The context is a strong predictor of negative action (see (3) tion see .combination of a person's self-regulating ability and the external regulatory features of the context, with four types of interactions. Self-regulated learning, therefore, may be explained and predicted by an individual's self-regulation in conjunction with the external characteristics of the context. Consequently, the prediction of the model is that the 1st combination should produce few positive and many negative emotions, high burnout and low engagement. The 2nd combination should produce medium-low positive emotions and negative medium-high, medium-high burnout and medium-low engagement. The 3rd combination should produce medium/high positive emotions and low negative emotions, medium-high engagement and medium-low burnout. The 4th combination should produce high positive emotions and low negative emotions, high engagement and low burnout and external mediation (favoring or discouraging regulation). Human learning is thus envisioned as the nout see .objectives: (1) to improve the heuristic technique for assessing the type of combination\u2014as established by SRL vs. ERL Theory\u2014using five types or levels; (2) to establish whether these interaction levels determined positive and negative achievement emotions, as defined in Pekrun's model; (3) to analyze whether there was a similar impact in the correlates of engagement and burnout. Hypotheses consistent with these objectives were defined as follows: (1) The possible combinations of student's level of self-regulation and level of external regulation offered by the teaching can be ordered in five progressive levels ; (2) the gradual increase of interaction level, ranging from 1 to 5, will lead to a proportionate increase in positive emotionality and a decrease in negative emotionality, as conceptualized by the Pekrun model; (3) using the same logic, these levels will correspond to a progressive increase in student engagement and a decrease in burnout.Based on the foregoing models and empirical data, this investigation had several Personal Self-Regulation (SR), and Regulatory Teaching (RT), we used a total sample of 440 undergraduate students from two universities in Spain. A selected sample of 336 students was used to analyze the type of combination. The sample was composed of students enrolled in Psychology, Primary Education, and Educational Psychology degree programs; 86.5% were women and 13.5% were men. Their ages ranged from 19 to 49, with a mean age of 23.08 (\u03c3X = 4.4) years.For the interdependence relations among low-medium-high levels of Short Self-Regulation Questionnaire (SSRQ) and for the factors of goal setting-planning (\u03b1 = 0.79), decision making (\u03b1 = 0.72) and learning from mistakes (\u03b1 = 0.72). Correlations have been studied, between each item and its factor total, among the factors, and between each factor and the complete questionnaire, with good results in all cases, except for the decision-making factor, which had a lower correlation with other factors (range: 0.41\u20130.58). The correlations between the original version and the complete version, and between the original and the short versions with a Spanish sample (complete SRQ with 32 items and short SRQ with 17 items) are better for the short version than for the complete version .This variable was measured using the Regulatory Teaching is Dimension 1 of the confirmatory model. IATLP-D1 comprises 29 items structured along five factors: Specific regulatory teaching, regulatory assessment, preparation for learning, satisfaction with the teaching, and general regulatory teaching. The scale was validated in university students and adequate internal consistency . The ATLP is a self-report instrument completed by the teacher and the students, available in Spanish and English versions. It also includes a qualitative part where students can make recommendations for improving each of the processes evaluated. As for the instrument's external validity, results are also consistent, since there are different interdependent relationships among perceptions of variables that exist in an academic environment.The Scales for Assessment of the Teaching-Learning Process, ATLP, student version measure the following eight emotions: class-related enjoyment, hope, pride, anger, anxiety, shame, hopelessness, and boredom. The learning-related emotions scale (LRE) contains 75 items and measures the same eight emotions in study situations. The test emotions scale (TES) contains 77 items that assess test-related enjoyment, hope, pride, relief, anger, anxiety, shame, and hopelessness. Each section is formed by three blocks of items, for assessment of emotions experienced either before, during, or after the achievement situations addressed in that section. Trait achievement emotions are assessed, that is, the student's typical, individual emotional reactions in achievement situations. The AEQ instructions can be altered for the purpose of measuring emotions experienced in a particular class subject (course-specific emotions), or in specific situations at a specific time (state achievement emotions).The Achievement Emotions Questionnaire, AEQ and five negative emotions . Two main criteria were used to decide which emotions to include. First, the emotions identified are frequently experienced by college students Pekrun, . Second,The three main types of achievement situations at university\u2014attending class, studying, and taking tests and exams\u2014differ according to function and social structure. This implies that emotions toward these situations would also differ. Enjoyment of classroom instruction, for example, would differ from enjoying the challenge of an exam. Some students may feel excited about going to class, others when taking exams. The AEQ takes this into account by providing separate scales for emotions that are class-related, learning-related, and test-related.Class-Related Emotions . The Cronbach alpha for this sample was 0.904, 0.803 (40 items), and 0.852 (40 items), for each part, respectively (80 items).(1) Learning-Related Emotions . The Cronbach alpha for this sample was 0.930, 0.880 (38 items), and 0.846 (37 items), for each part, respectively (75 items).(2) Test-Related Emotions . The Cronbach alpha for this sample was 0.913, 0.824, and 0.869, for each part, respectively (77 items).(3) This version has shown adequate reliability and construct validity indices in a cross-cultural study.Engagement was assessed with a validated Spanish version of the Utrecht Work Engagement Scale for Students . The Cronbach alpha for this sample was 0.900 (14 items), 0.856 (7 items), and 0.786 (7 items), for each part, respectively.Burnout was assessed with a validated Spanish version of the Burnout Scale for Students . The Cronbach alpha for this sample was 0.874 (15 items), 0.853 (8 items), and 0.793 (7 items), for each part, respectively.platform .Participants voluntarily completed the scales using an online A previous confirmatory factor analysis was conducted in this sample as evidence of factorial validity and to ensure the previous structural fit of each inventory , using the statistical program AMOS (v. 22) Reliability was also calculated (Cronbach Alpha) through SPSS (v.25).Using an ex-post-facto design, first, a 3 K-means cluster analysis was conducted to establish low-medium-high groups in each of the two variables: Personal Self-Regulation (SR) and Regulatory Teaching (RT). In the case of the SR variable, the values formed the centers of the clusters, response ranges being low (1.00\u20133.09), medium (3.10\u20133.84), and high (3.85\u20135.00). In the case of the RT variable , formed the centers of the clusters, response ranges being low (1.00\u20132.34), medium (2.35\u20132.83) and high (2.84\u20135.00). In addition, several ANOVAs and MANOVAs were carried out, to ascertain the effect of low-medium-high levels of the dependent variable, achievement emotions. Also, using a 3-factor design (low-medium-high self-regulation levels) \u00d7 3 (low-medium-high levels of regulatory teaching), several MANOVAs were conducted, taking the aforementioned levels as the independent variable. Finally, based on the low-medium-high groups in both variables (SR and RT), five combinations were configured, according to the theoretical model proposed see . MANOVAsSR IV (low-medium-high levels) , and RT IV (low-medium-levels) , was noted on the CAE. The statistically significant partial effect was maintained of the SR IV (low-medium-high levels) for both Positives Emotions , and Negatives Emotions . The statistically significant partial effect was maintained of the PR IV (low-medium-high levels) for both Positive Emotions , and Negative Emotions . No statistical effect of significant interaction appeared.A statistically significant main effect of the SR IV (low-medium-high levels) , and RT IV (low-medium-levels) , was noted on the factors of CAE. The statistically significant partial effect was retained for enjoyment , for hope , for pride , for boredom , for anger , for anxiety , for shame , and for hopelessness .Complementarily, a statistically significant main effect of the SR IV (low-medium-high levels) , and RT IV (low-medium-levels) , was noted on the LAE. The statistically significant partial effect was maintained of the SR IV (low-medium-high levels) for both Positive Emotions , and Negative Emotions . The statistically significant partial effect was maintained of the PR IV (low-medium-high levels) for both Positive Emotions , and Negative Emotions . No statistical effect of significant interaction appeared.A statistically significant main effect of the SR IV (low-medium-high levels) , and RT IV (low-medium-levels) , was noted on the factors of LAE. The statistically significant partial effect of SR IV was retained for enjoyment , for hope , for pride , for boredom , for anger , for anxiety , for shame , and for hopelessness . A statistically significant partial effect of RT IV was retained for enjoyment , for hope , for pride , for boredom , for anger , for anxiety , for shame , and for hopelessness .Complementarily, a statistically significant main effect of the SR IV (low-medium-high levels) , and RT IV (low-medium-levels) , was noted on the TAE. The statistically significant partial effect was maintained of the SR IV (low-medium-high levels) for both Positive Emotions , and Negative Emotions . The statistically significant partial effect was maintained of the PR IV (low-medium-high levels) for both Positive Emotions , and Negative Emotions . No statistical effect of significant interaction appeared.A statistically significant main effect of the SR IV (low-medium-high levels) , and RT IV (low-medium-levels) , was noted on the factors of TAE. The statistically significant partial effect was retained for enjoyment , for hope , for pride , for relief , for anger , for anxiety , for shame , and for hopelessness .Complementarily, a statistically significant main effect of the Self-Regulation IV (low-medium-high levels) , and Regulatory Teaching IV (low-medium-high levels) was observed on Engagement-Burnout levels. The statistically significant partial effect was maintained of Self-Regulation IV both Engagement , and Burnout . A statistically significant general main effect of the Regulatory Teaching IV (low-medium-high levels) both Engagement-Burnout .A statistically significant general main effect of the Self-Regulation IV's effect (low-medium-high levels) on the components of engagement-burnout yielded a statistically significant main effect . The statistically significant partial effect was retained for vigor , for dedication , for absorption , for exhaustion , for cynicism , for lack of effectiveness .The combined analysis of the Regulatory Teaching IV (low-medium-high levels) was observed on the components of engagement-burnout levels . The statistically significant partial effect was retained for vigor , for dedication , for absorption , for exhaustion , for cynicism , for lack of effectiveness showed a statistically significant main effect of the five interaction types on the low-medium-high levels of SR and of RT see :Combination 1 presented a statistically significant low level in SR and low level in RT (1 and 1). The average regulation level of 1.0, and the rank level is 1. The range of regulation tends toward low SR and low RT, associated with a high level of dysregulation. The most probable emotions are low levels of positive emotions and high levels of negatives emotions. Consequently, the effects are a high level of stress: high burnout and low engagement.Combination 2 had a statistically significant low-medium level in SR and medium-low level in RT and vice versa . The average regulation level is 1.5, and the rank level is 2. The range of regulation tends toward low-medium SR and low-medium RT, and vice versa, associated with medium-low level of dysregulation. The most probable emotions are medium-low level of positive emotions and medium-low level of negative emotions. Consequently, the effects are a medium-high level of stress: medium-high burnout and medium-low engagement.Combination 3 presented a statistically significant medium SR level (2) and medium RT level (2 and 2). The average regulation level of 2.0, and the rank level is 3. The range of regulation tends toward medium SR and medium RT, associated with medium level of dysregulation. The most probable emotions are medium level of positive emotions and medium level of negative emotions. Consequently, the effects are a medium level of stress: medium burnout and medium engagement.Combination 4 had a statistically significant medium SR- high RT and high RT- medium SR . The average regulation level is 2.5, and the rank level is 4. The range of regulation tends toward high SR\u2013medium RT and medium SR and high RT, associated with a good level of regulation. The most probable emotions are medium-high level of positive emotions and medium-low level of negative emotions. Consequently, the effects are a medium-low level of stress: medium-low burnout and medium-high engagement.Combination 5 presented statistically significant high SR- high RT and high RT- high SR (3 and 3). The average regulation level is 3.0, and the rank level is 5. The range of regulation tends toward high SR\u2013high RT, associated with a high level of regulation. The most probable emotions are high level of positive emotions and low level of negative emotions. Consequently, the effects are a low level of stress: low engagement and high burnout.The MANOVA produced statistically significant differences among the five groups in levels of self-regulation (SR) and regulatory teaching (RT); both variables were adequately configured as established in five combinations of SR and RT as IV was noted in Class Achievement Emotions (CAE), Learning Achievement Emotions (LAE) and Test Achievement Emotions (TAE). The statistically significant partial effect was maintained of the five combinations of SR and RT IV for both Positive Emotions and Negative Emotions. In the case of positive emotions, a significant statistical effect appeared in favor of higher levels , while for negative emotions the effect was reversed, in favor of lower levels . The statistically significant partial effect was maintained for each positive emotion , and for negative emotions . Complementarily, in the case of engagement, a significant statistical effect appeared in favor of higher levels , while for burnout the effect was reversed, in favor of lower levels . The statistically significant partial effect was maintained for engagement factors , and for burnout factors see . The graself-regulation and the level of external regulation offered by the teaching process. Furthermore, this type of interaction can be understood as the combination of low-medium-high levels of both factors, as seen in prior evidence and the regulatory level of the teaching process (low-medium-high), along a continuum. This allows for an improved combination model that organizes this interactive reality, as compared to the prior version of this theoretical model and engagement, and a decrease in negative emotions , deactivation (boredom and relief) and burnout. On one hand, this lends empirical support to the construct of self-regulation, by showing that it has the potential to discriminate degrees of positive and negative emotions in students. This result is consistent with plentiful prior evidence that has shown a positive, significant correlation between self-regulation and the personality factor of conscientiousness, leading us to consider that self-regulation is a meta-behavioral variable that materializes this personality variable, associated with less stress, in contrast to the variable of neuroticism molar psycho-educational models in real settings, and not only knowledge about relations between achievement emotions and personality variables, in molecular-level models or dysregulated behaviors (high level of negative and low level of positive emotions), university guidance and counseling services ought to detect and help these types of students, as they begin their university studies, to promote stress management and coping strategies, and so minimize the impact of negative effects from the university experience. Certain programs in current use might help toward this end and of negative emotionality have been amply associated with academic failure and dropout (Putwain et al., The datasets generated for this study are available on request to the corresponding author.This studies involving human participants were reviewed and approved by Comit\u00e9 de \u00c9tica de la Investigaci\u00f3n. Universidad de Navarra. The patients/participants provided their written informed consent to participate in this study.JF has coordinated the R&D Project, has made the general design, data analysis, and first writing of the manuscript. JM-V has reviewed the design and analysis of data. FP-S has collected the data sample and has revised the manuscript. AG-U has collected the data sample and has revised the manuscript. MV has reviewed the previous evidence and the theoretical foundation. PP has provided the translation of the instruments and validated them in Spanish.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "Over the past decade, the identification of potential genetic and pharmacological modifiers of lifespan and age-related pathologies in C. elegans and other model organisms has yielded fruitful leads for follow-up investigation. A major limitation of such studies, however, is that they are often time-consuming and labor-intensive. The advent of affordable high-quality digital cameras, robotics systems, and 3D printers, as well as the decreasing costs of image storage and processing have allowed us to automate data capture and analysis at an unprecedented scale. To this end, our group developed a tool consisting of an unbiased, high-throughput, automated robotic system to perform genetic and pharmacological quantification of lifespan and health measures in C. elegans and related nematode species. The WormBot utilizes industry-standard, commercially available robotics components to position a digital camera over individual wells of standard 12-well culture plates, containing a small population of C. elegans per well. A high-resolution image is captured of each plate every 10 minutes throughout the course of the experiment. Our software processes the images for stabilization, compiles them into a time-lapse series for each well, and quantifies survival and mobility with minimal input. In addition, a short video is captured of each well once each day, to allow for quantitative analyses of activity and coordinated movement. We will describe this technology and present applications to screen genetic and pharmacological libraries in aging and age-related disease."} +{"text": "There is limited information on the epidemiology and treatment patterns of rheumatoid arthritis (RA) across the Arab region. We aim in this study to describe the demographic characteristics, clinical profile, and treatment patterns of patients of Arab ancestry with RA.This is a cross sectional study of 895 patients with established rheumatoid arthritis enrolled from five sites , and United Arab Emirates). Demographic characteristics, clinical profile, and treatment patterns are compared between the five countries.The majority of our patients are women, have an average disease duration of 10 years, are married and non-smokers, with completed secondary education. We report a high (>80%) ever-use of methotrexate (MTX) and steroids among our RA population, while the ever-use of disease modifying anti-rheumatic drugs (DMARDs) and TNF-inhibitors average around 67% and 33%, respectively. There are variations in RA treatment use between the five country sites. Highest utilization of steroids is identified in Jordan and KSA , while the highest ever-use of TNF-inhibitors is reported in KSA .Disparities in usage of RA treatments among Arab patients are noted across the five countries. National gross domestic product (GDP), as well as some other unique features in each country likely affect these. Developing treatment guidelines specific to this region could contribute in delivering standardized therapies to RA patients. The presability , with a sability . Other dsability \u201311.In the Middle East and North Africa (MENA) region, the epidemiology of RA remains poorly understood with a dearth of data on its prevalence and disease activity among Arab populations. A recent global burden study estimated RA prevalence in MENA region as among the lowest at 0.16% \u201315, RA dDisease modifying anti-rheumatic drugs (DMARDs) are the mainstay treatment for RA prescribed to relieve joint pain and swelling, and to reduce disease activity and disability , 17. NewIn the context of limited data on the epidemiology of RA and its treatment patterns in the MENA region, we aim in the study to describe the demographic, clinical characteristics, and treatment patterns of patients with RA of Arab ancestry living in five Middle Eastern countries.This is a secondary data analysis of a cross-sectional study of RA patients enrolled in the Genetics of Rheumatoid Arthritis in Some Arab States (GRASS) multicenter case-control study . Data weAll the enrolled patients were adequately informed about the research and provided written consent prior to study enrollment. The study was approved by the ethics boards at all study sites. These include Jordan Hospital Institutional Review Board (Jordan), King Faisal Specialist Hospital and Research Center Institutional Review Board (KSA), American University of Beirut Institutional Review Board (Lebanon), Hamad Medical Center Institutional Review Board and Weill Cornell Medicine-Qatar Institutional Review Board (Qatar), and Dubai Scientific Research Ethics Committee (UAE). A questionnaire was completed by all patients and their physicians capturing sociodemographic and clinical data, including medical history of RA, limited comorbidities and medication use.Sociodemographic characteristics collected included self-reported ancestry, nationality, age, sex, education, and marital status. Self-reported ancestry was classified as Gulf, Levant, or Africa if three or more of the patients\u2019 grandparents originated from the Arabian Gulf, Levant, or Africa regions, respectively; all other reported ancestry were grouped in the \u201cother/mix\u201d classification. Education was categorized into three groups: patients who completed primary education or none, middle or secondary school, and college or university education. Marital status was categorized as either never- or ever-married. Data on smoking status, frequency and duration (in years) for both cigarettes and shisha (narghileh) were collected and classified as never- or ever-smoked.Information on age at diagnosis, duration of disease, presence of rheumatoid factor (RF) and /or anti-cyclic citrullinated peptide , presence of comorbidities ), and the use of medications to treat RA were collected from the patients\u2019 medical records. The use of medications was described as never- or ever-use for the following medications: methotrexate (MTX), DMARDs , steroids, and TNF-inhibitors and non-TNF-inhibitors (rituximab) biologics. The use of non-TNF-inhibitor biologics (rituximab) was minimal across all the sites and not included in the analysis. Information on other non-TNFi drugs (tocilizumab and abatacept) were not collected as these medications were not readily available in the countries when the study was initiated in 2012. This impeded us from systematically capturing such data.Sample characteristics were summarized using frequency distributions except for patients\u2019 age, age at diagnosis, and duration of disease for which the mean and standard deviation were calculated. Socio-demographic, smoking status, co-morbidities, and treatment patterns were compared between the five country sites using the Chi-square and Fisher\u2019s exact tests. One way Analysis of Variance (ANOVA) was used to compare patients\u2019 age, age at diagnosis, and duration of disease between the five country sites. For each of the different types of medication, bivariate and then multivariate logistic regressions were built to assess what factors were associated with their use. Unadjusted and adjusted odds ratios (OR) were reported along with their respective 95% confidence intervals (CI). Since UAE did not collect information on anti-CCP and RF, we conducted a sensitivity analysis excluding the UAE sample to factor anti-CCP and RF in the regression. Significance was defined at the 5% level. All analyses were done using IBM-SPSS version 24.0 .A total of 895 patients with RA were enrolled from the five participating sites. The majority were female (85%), ever-married (87.5%), and with at least middle or secondary school education (73%). About 42% and 39% reported the Gulf and Levant as their ancestry, respectively. The baseline characteristics are summarized in Across country sites, significant variability was noted in self-reported ancestry, disease duration, education, marital status, cigarette and shisha smoking, and presence of T2DM . In LebaMethotrexate (87.8%) and steroids (80.6%) were the most commonly ever-used medications across all sites. Around two thirds of patients ever-used DMARDs other than MTX. Only one third of patients ever- used TNF-inhibitors. Additionally, there were significant differences in the types of medication used between countries . HighestAdjusted analysis showed tOur cross-sectional study examined socio-demographic, clinical, and treatment profile of 895 RA patients of Arab ancestry recruited from five countries in the Middle East. The majority are women, have an average disease duration of 10 years, are married and non-smokers, with completed secondary education, and comorbidities consistent with what has been described in other populations in the world , 24. OurWe observed a high ever-use of MTX and steroids among our patients with little variation across all centers, while the ever-use of DMARDs (other than MTX) and TNF-inhibitors significantly varied. This high prescription of MTX is a substantial improvement from prior reports from theThe significant variations in the use of TNF-inhibitors amongst the sites is likely to be explained by the differences in the healthcare systems as well as patient factors. Total health expenditure per capita ranges from as low as USD 257 in Jordan to as high as USD 2,030 in Qatar and acceWe established factors associated with the choice of RA treatment in our cohort. Increasing age of diagnosis is inversely associated with use of MTX in parallel to the pattern reported from the UK and SwedThere are biases inherent to a cross-sectional analysis of a convenience sample. The prevalence of co-morbidities and medication use could be underestimated due to recall bias as the information could be missing from the patients\u2019 medical records. Records were not maintained for eligible patients who refused participation, thus we are unable to evaluate the characteristics of those who refused to participate. In addition, there was a lack of a central lab to test for anti-CCP and RF to ensure the accuracy of those results. The missing data on anti-CCP and RF status precluded a complete clinical assessment of RA in our study population.Despite these limitations, our study fills major gaps on RA epidemiology and characteristics pertaining to RA patients of Arab ancestry in the Middle East. Also, the novelty of our study lays in the description, investigation, and comparison of medication patterns between five nations of different GDP levels in the Eastern Mediterranean region.In summary, this is the largest study of RA patients from the Middle East. Notable findings include younger age of onset of the disease and significant variations in treatment patterns amongst countries. Future studies should aim to address disease severity, remission rates as well as patient and physician treatment preferences. Lastly, developing region specific guidelines for RA treatment, could result in delivering equitable care to patients.S1 File(SAV)Click here for additional data file."} +{"text": "An increasing number of families, funders, and community providers seek very brief psychosocial caregiver interventions, yet the evidence for such condensed interventions is not established. Based on the Savvy Caregiver Program, we explored the feasibility, acceptability, and outcome trends for a condensed 3-session version titled, Savvy Express. Based on a single-group, pre- and post-test intervention design, we examined post-intervention and 3-month data on 116 English-speaking racially and ethnically diverse care partners caring family members with Alzheimer\u2019s disease and related dementias. 41% of the sample was non-Latino white and comprised of Latinas, African Americans and Asian American/API. Most care partners were either adult children or spouses caring for someone with AD or other dementia. Over 80% were college educated. Two of three participants completed all 3 classes. Our findings indicate significant improvements in caregiver levels of depressive symptomatology and anxiety, competence, management of the situation, reduction of expectations, making positive comparisons, and reactivity to the family member\u2019s memory behavior. Upwards of 90% would recommend the program to other caregivers. Savvy Express is a brief caregiver intervention with high acceptability and feasibility. Improvements in care partner psychosocial outcomes signal a promising practice to reduce the burden of caregiving. A major focus of the paper focuses on barriers and facilitators to uptake of the study procedures and intervention with community-based partners. Future work is needed to establish the efficacy of Savvy Express across a longer observation period, and with less educated, low-income participants, and limited English-speaking families."} +{"text": "We conducted a detailed analysis of trends in new HIV diagnoses in Australia by country of birth, to understand any changes in epidemiology, relationship to migration patterns and implications for public health programs.Poisson regression analyses were performed, comparing the age-standardised HIV diagnosis rates per 100,000 estimated resident population between 2006\u20132010 and 2011\u20132015 by region of birth, with stratification by exposure . Correlation between the number of permanent and long-term arrivals was also explored using linear regression models.Between 2006 and 2015, there were 6,741 new HIV diagnoses attributed to male-to-male sex and 2,093 attributed to heterosexual sex, with the proportion of diagnoses attributed to male-to-male sex who were Australian-born decreasing from 72.5% to 66.5%. Compared with 2006\u20132010, the average annual HIV diagnosis rate per 100,000 in 2011\u201315 attributed to male-to-male sex was significantly higher in men born in South-East Asia (summary rate ratio (SRR) = 1.37, p = 0.001), North-East Asia and the Americas , but significantly lower as a result of heterosexual sex in men born in South-East Asia , Southern and Central Asia and Sub-Saharan Africa and women born in South-East Asia and Sub-Saharan Africa . Positive associations were observed between the number of permanent and long-term arrivals and HIV diagnoses particularly in relation to diagnoses associated with male-to-male sex in men from North Africa and the Middle East, North Asia, Southern and Central Asia and the Americas.The epidemiology of HIV in Australia is changing, with an increase in HIV diagnosis rates attributed to male-to-male sex amongst men born in Asia and the Americas. Tailored strategies must be developed to increase access to, and uptake of, prevention, testing and treatment in this group. Migrants in high income, low prevalence countries can be disproportionately affected by HIV, with higher diagnosis rates and a greater likelihood of being diagnosed late, with a CD4 count under 350 cells/ul \u20133. HistoAustralia has a concentrated HIV epidemic, with around three quarters of new diagnoses each year attributed to male-to-male sex and one-fifth to heterosexual exposure . The numMigrants with HIV in Australia have differing histories and reasons for migration. Understanding this diversity is important in designing programs which maximise access to HIV testing, treatment and care to reduce HIV transmission. Australia has committed to working towards elimination of transmission of HIV by 2020 , and addIn this study, we sought to determine if, similar to other high-income countries, the epidemiology of HIV by country of birth and exposure in Australia has changed over time, and if any changes are related to migration patterns. Findings are intended to inform HIV programs for people born overseas.Under procedures described previously , all newData extracted from the National HIV Registry for this analysis were year and age at diagnosis, sex, country of birth (routinely collected from 2002), mode of exposure to HIV ; MSM with dual risk of injecting drug use (IDU); heterosexual sex; and other/undetermined exposure, including mother-to-child transmission, direct blood/tissue exposure, and iatrogenic exposure, likely place of acquisition and year of arrival in Australia.Countries of birth were grouped into one of eight regions based on categories used by the Australian Bureau of Statistics (ABS) . The ABSThe likely place of acquisition and year of arrival variables have only been available nationally since 2015, so analysis of these variables was undertaken using data from this year alone.Data on estimated resident population (ERP) and permDesk-based, secondary data analysis was conducted. All people newly diagnosed with HIV in Australia between 1 January 2006 and 31 December 2015 with the HIV exposure recorded as male-to-male or heterosexual sex were included. Excluded cases had a prior diagnosis overseas, no country of birth was recorded, and/or were transgender, the last exclusion to allow stratification of risk by sex.The study period was divided into two equal five-year intervals for comparison of changes over time\u2013(i) 2006 to 2010 and (ii) 2011 to 2015. Descriptive analysis was conducted to calculate the proportions of HIV diagnoses by region of birth, by sex and mode of exposure . The Wilcoxon rank sum test was used to compare median age at diagnosis by region of birth and exposure, using people born in Australia as the comparison. Pearson\u2019s chi-squared test was used to compare overseas-born diagnoses by likely place of acquisition and reported exposure.Average annual HIV diagnosis rates per 100,000 population were calculated by region of birth for each time period, with stratification by exposure , and age standardisation using the 2001 ABS standard population as a refThe difference in the average annual age-standardised HIV diagnosis rate between 2006\u20132010 and 2011\u20132015 for each region of birth was analysed using univariate Poisson models, with number of diagnoses as the outcome and time interval as the independent variable. Summary rate ratios (SRRs) for each region of birth and exposure category were calculated, using the 2006 to 2010 time period as the reference.To understand the relationship between migration patterns and new HIV diagnoses, we used linear regression models for each region of birth and mode of exposure, with number of HIV diagnoses per year as the dependent variable and permanent and long-term arrivals as the independent variable. Models were adjusted by including age as a covariate in the multivariate analysis. This analysis provides a more sensitive measure of any relationship between migration patterns and HIV diagnoses than using ERP alone, as ERP is based on population projections whereas permanent and long-term arrivals are real-time data.For all analyses, the population denominator used for calculation of HIV diagnosis rates in men\u2014whether attributed to male-to-male sex or heterosexual sex\u2014was the overall male population, as population numbers stratified by mode of exposure are not available. This method is consistent with those used in producing national surveillance estimates .All analyses were performed using STATA IC 14 , with the significance level set at p>0.05.Ethical approval was received from the University of New South Wales Human Research Ethics Committee (Reference HC16482).Between 2006\u20132010 and 2011\u20132015, the proportion of HIV diagnoses attributed to male-to-male sex which were in men decreased from 72.5% to 66.5%, while the proportion born in South-East, North-East or Southern and Central Asia increased from 9.7% to 15.8%. The proportion attributed to heterosexual sex which were in Australian-born men and women increased from 33.7% to 42.5% while the proportion born in Sub-Saharan Africa and South-East Asia decreased from 27.2% to 18.2%, and 15.8% to 12.3% respectively.Age at diagnosis is an important factor in understanding who is at risk and for targeting of prevention, treatment and care programs.For diagnoses attributed to male-to-male sex, the median age at diagnosis compared to Australian-born men was significantly lower in both time periods in South-East Asian and North-East Asian born men, higher for men born in North-West Europe, and lower in 2011\u20132015 alone for men born in the Americas. For diagnoses attributed to heterosexual exposures, the median age at diagnosis compared to the Australian-born was significantly lower in both time periods in men and women born in Oceania, South-East Asia, Southern and Central Asia and Sub-Saharan Africa and higher in those born in North-West Europe. The median age at diagnosis compared to Australian-born was higher in 2006\u20132010 alone for those born in Southern and Eastern Europe and lower in 2011\u20132015 alone for men and women born in North Africa and the Middle East and North-East Asia. There were no significant differences in age for other region of birth or exposure groupings compared to Australian-born, in either 2006\u20132010 or 2011\u20132015.In 2006\u20132010, amongst HIV diagnoses attributed to male-to-male sex, the highest proportion were among Australian-born men, followed by men born in South-East Asia, North-West Europe, Oceania, the Americas, North-East Asia, Southern and Eastern Europe, Sub-Saharan Africa, North Africa and the Middle East and Southern and Central Asia. In 2011\u20132015, the highest proportion were among Australian-born men, followed by men born in South-East Asia, North-West Europe, North-East Asia, Oceania, the Americas, Southern and Eastern Europe, Southern and Central Asia, Sub-Saharan Africa, and North Africa and the Middle East.In 2006\u20132010, amongst HIV diagnoses attributed to heterosexual sex, the highest proportion were among Australian-born men and women, followed by those born in Sub-Saharan Africa, South-East Asia, North-West Europe, Oceania, North Africa and Middle East, Southern and Central Asia, Southern and Eastern Europe, the Americas and North-East Asia. In 2011\u201315, the highest proportion were among Australian-born men and women, followed by those born in Sub-Saharan Africa, South-East Asia, Oceania, North-West Europe, North Africa and Middle East, Southern and Central Asia, North-East Asia, Southern and Eastern Europe and the Americas.There were 383 people born overseas and diagnosed with HIV in 2015. Of these, data on time since arrival was available for 197 (51.4%) diagnoses, on likely place of acquisition for 297 (77.6%) diagnoses, and on both time since arrival and likely place of acquisition for 177 (46.2%) diagnoses. The median time since arrival was three years (IQR 1\u201311 years) for men in whom male-to-male sex was the likely exposure, six years (IQR 1\u201317 years) for men and five years (IQR 1\u201311 years) for women in whom heterosexual sex was the likely exposure.2 = 28.0, p<0.001).Of the 230 new HIV diagnoses attributed to male-to-male sex for whom data on likely place of acquisition was available, 78 (33.9%) were acquired overseas. The proportion acquired overseas was higher for HIV diagnoses attributed to heterosexual contact and the Americas compared with Australian-born men. Rates were significantly lower compared to Australian-born men in men born in Southern and Eastern Europe , North Africa and the Middle East and Southern and Central Asia , but not significantly different for men born in Oceania, North-West Europe, North-East Asia and Sub-Saharan Africa.In 2011 to 2015, age-standardised new HIV diagnosis rates attributed to male-to-male sex were significantly higher in men born in South-East Asia , North-East Asia and the Americas compared with men born in Australia. Rates were significantly lower in men born in Southern and Eastern Europe , North Africa and the Middle East and Southern and Central Asia . Rates were not significantly different for men born in Oceania, North-West Europe and Sub-Saharan Africa.Between the time periods 2006\u20132010 and 2011\u20132015, HIV diagnosis rates attributed to male-to-male sex increased 1.37 times (95% CI 1.14\u20131.65) in men born in South-East Asia, 2.18 times (95% CI 1.64\u20132.89) in men born in North-East Asia and 1.37 times (95% CI 1.04\u20131.79) for men from the Americas, as expressed by the SRR. Changes for other regions of birth were not significant.In 2006 to 2010, age-standardised HIV diagnosis rates attributed to heterosexual contact were significantly higher in men born in North-West Europe , North Africa and the Middle East , South-East Asia , Southern and Central Asia , the Americas and Sub-Saharan Africa compared to Australian-born men. Rates were lower or not significantly different for men born in Southern and Eastern Europe, North-East Asia or Oceania compared with men born in Australia.In 2011 to 2015, new HIV diagnosis rates attributed to heterosexual contact were significantly higher in men born in Oceania , North-West Europe , North Africa and the Middle East , South-East Asia , Southern and Central Asia , the Americas and Sub-Saharan Africa than in the Australian-born. Rates were not significantly different for men born in Southern and Eastern Europe or North-East Asia.Between the time periods 2006\u20132010 and 2011\u20132015, new HIV diagnosis rates attributed to heterosexual contact decreased significantly in men born in South-East Asia , Southern and Central Asia and Sub-Saharan Africa . Changes for other regions of birth were not significant.Age-standardised new HIV diagnosis rates attributed to heterosexual contact from 2006 to 2010 were higher than in women born in Oceania , North-West Europe North Africa and the Middle East , South-East Asia , the Americas and Sub-Saharan Africa than in Australian-born women. Rates were not significantly different for women from Southern and Eastern Europe, North-East Asia and Southern and Central Asia compared with those born in Australia.From 2011 to 2015, new HIV diagnosis rates attributed to heterosexual contact were significantly higher in women born in Oceania , Southern and Eastern Europe , North Africa and the Middle East , South-East Asia and Sub-Saharan Africa than in Australian-born women. Rates were not significantly different for other regions of birth.Between 2006\u20132010 and 2011\u20132015, HIV diagnosis rates attributed to heterosexual contact decreased significantly in women born in South-East Asia and Sub-Saharan Africa, by 0.61 (95% CI 0.45\u20130.83) and 0.61 (95% CI 0.48\u20130.79) times respectively. Changes for other regions of birth were not significant.After adjusting for age, HIV diagnoses attributed to male-to-male sex had a significant positive association with permanent and long-term arrivals for those from North Africa and the Middle East, North-East Asia and the Americas, and a borderline significant positive association for those from Southern and Central Asia . For malThe epidemiology of HIV in Australia is changing. While HIV diagnosis rates for Australian-born men and women in all exposure categories have been stable, rates attributed to male-to-male sexual exposure among men born in South-East Asia, North-East Asia and the Americas increased significantly between 2006\u20132010 and 2011\u20132015, and are higher than those of Australian-born men. In 2011\u20132015 Asian-born men accounted for over 15% of all new HIV diagnoses attributed to male-to-male sex in Australia. Diagnosis rates attributed to heterosexual exposure amongst men and women born in South-East Asia and Sub-Saharan Africa, and men born in Southern and Central Asia, decreased over the ten years but remained higher than rates in Australian-born people.Migration patterns and the epidemiology of HIV in countries of origin may partially explain these changes. With respect to male-to-male sexual exposure, increases in the number of new arrivals were significantly correlated with increases in new HIV diagnoses for those from North Africa and the Middle East, North-East Asia, Southern and Central Asia and the Americas. In relation to heterosexual exposure, increases in new arrivals were associated with increases in new HIV diagnoses in men from Sub-Saharan Africa, and decreases in new HIV diagnoses for men and women from South-East Asia.These trends are consistent with the presence of concentrated epidemics among men who have sex with men (MSM) in countries in North-East Asia, Southern and Central Asia and the Americas; and with changes in South-East Asian countries such as Thailand and Cambodia from generalised to concentrated epidemics amongst populations such as MSM . FindingA positive association between new arrivals and new HIV diagnoses does not mean people born overseas acquired HIV prior to migration. Data for 2015 suggest overseas-born men diagnosed with HIV attributed to male-to-male sex are more likely overseas-born men and women diagnosed with HIV attributed to heterosexual sex to have acquired the infection in Australia. Recent European studies also show overseas-born MSM diagnosed with HIV are now more likely to have acquired the infection post migration . More reAsian-born men diagnosed with HIV may have recently arrived in Australia , potentiThe strength of this study is the use of population-wide data on new HIV diagnoses and new arrivals, and high level of completeness of key variables such as country of birth in the National HIV Registry. Limitations include first, the use of the overall male population as the denominator in analyses for diagnoses in men attributed to male-to-male sex, as there are no population estimates for MSM disaggregated by region of birth. HIV diagnosis rates attributed to male-to-male sex across all regions of birth have therefore been underestimated. The analysis of association with migration patterns also assumes MSM from different regions of birth are as likely to migrate as are heterosexual men.Secondly, the National HIV registry does not collect information on visa status, so we could not disaggregate cases by length of stay or visa category. Data on place of acquisition and year of arrival was only available for 2015, and incomplete. Data from future years on these variables is needed to verify the findings. Finally, we purposely chose to present data by region rather than country of birth to avoid stigmatising particular communities, however this analysis may mask differences within a given region. Numbers for some regions such as the Southern and Central Asia remain small, so findings should be interpreted with caution.Further research is needed into HIV risk amongst Asian-born MSM, their social and sexual networks, behavioural and cultural factors which might increase risk amongst different sub-groups and communities, and factors influencing the uptake of HIV prevention, testing and treatment measures, to better tailor interventions for this group. Existing evidence suggests integration of provider-initiated testing and counselling into primary care is a promising approach , as is eThe epidemiology of HIV amongst overseas-born populations in Australia is complex and changing. HIV programs must adapt to emerging groups at risk, such as Asian-born MSM, while still effectively targeting groups such as South-East Asian and Sub-Saharan African born heterosexual men and women who continue to experience high HIV diagnosis rates."} +{"text": "Scientific Reports 10.1038/s41598-018-26374-9, published online 21 May 2018Correction to: The Acknowledgements section in this Article is incomplete.\u201cWe thank the TCGA project, which generated comprehensive, multi-dimensional maps of key genomic changes in liver hepatocellular carcinoma. This work was supported by grants from the Science and Technology Department of Sichuan Province, No. 30504010361 and No. 2014FZ0123 and grants from the National Natural Science Foundation of China, No. 81571565.\u201dshould read:\u201cWe thank the TCGA project, which generated comprehensive, multi-dimensional maps of key genomic changes in liver hepatocellular carcinoma. This work was supported by grants from the Science and Technology Department of Sichuan Province, No. 30504010361 and No. 2014FZ0123, grants from the National Natural Science Foundation of China, No. 81571565 and grants from the Health and Family Planning Commission of Sichuan Province, No. 2015SZ0241.\u201d"} +{"text": "The Funding statement is incorrect. The correcting Funding statement is: This study was supported by FAPES- Espirito Santo Research and Innovation Support Foundation, Processos: 54690722/2011 and 84324600/2018; CNPq; UFES. The funders did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Mass spectrometry-based proteomics has had a formidable development in recent years, increasing the amount of data handled and the complexity of the statistical resources needed. Here we present SanXoT, an open-source, standalone software package for the statistical analysis of high-throughput, quantitative proteomics experiments. SanXoT is based on our previously developed weighted spectrum, peptide and protein statistical model and has been specifically designed to be modular, scalable and user-configurable. SanXoT allows limitless workflows that adapt to most experimental setups, including quantitative protein analysis in multiple experiments, systems biology, quantification of post-translational modifications and comparison and merging of experimental data from technical or biological replicates.https://wikis.cnic.es/proteomica/index.php/SSP.Download links for the SanXoT Software Package, source code and documentation are available at jvazquez@cnic.es or ebonzon@cnic.esBioinformatics online. The Centro Nacional de Investigaciones Cardiovasculares Carlos is supported by the Spanish Ministry of Economy, Industry and Competitiveness (MEIC) and the Pro-CNIC Foundation, and is a Severo Ochoa Center of Excellence (MEIC award SEV-2015-0505).Conflict of Interest: none declared.Supplementary InformationClick here for additional data file."} +{"text": "The first two authors, Wiwat Chancharoenthana and Asada Leelahavanichkul, should be noted as equal co-corresponding authors.There is an error in the Funding statement. The correct Funding statement is as follows: All of the funding supports during study period including the Development of New Faculty Staff fund and Ratchadapiseksomphot Endowment Fund 2017 (76001-HR), Faculty of Medicine, Chulalongkorn University and National Science and Technology Development Agency (NSTDA: P-13-00505)\u2013Dr. Asada Leelahavanichkul. A.L. is under Center of Excellence in Immunology and Immune Mediated Diseases group. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.The publisher apologizes for the errors."} +{"text": "A grant is missing from the Funding statement. This work was also supported by a National Institutes of Health grant (R01DK108088 to LD).The full, amended Funding statement is included below:http://www.kumc.edu/kumcri/sponsored-programs-administration/internal-funding-opportunities/lied-basic-science-grant-program.html to LD. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. University of Kansas Medical Center KINBRE Bridging Grant FY2015-16 to LD. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. National Institute of General Medical Sciences https://www.nigms.nih.gov/Pages/default.aspx (grant number P20GM103549 and P30GM118247) to KUMC Department of Pharmacology, Toxicology, and Therapeutics. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.National Institutes of Health grant number R01DK108088 to (LD). The funders had no role in study design, data collection, analysis, decision to publish, or preparation of this manuscript. This content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. University of Kansas Medical Center Lied Pilot Project Grant FY2014"} +{"text": "Spiroplasma in western group of populations of H.axyridis was supported by the Russian Foundation for Basic Research, 15-04-07466_A (IG). The study of Spiroplasma in eastern group of populations of H.axyridis\u2014the source of the global invasion, was supported by the Russian Science Foundation, 16-16-00079 (IZ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.There are errors in the Funding section. The publisher apologizes for the errors. The correct funding information is as follows: The study of"} +{"text": "The emerge of nanotechnology along with the success of the microelectronics industry has motivated the miniaturization of biosensors into the nano/microscale. This Special Issue highlights recent advances in microscale and nanoscale biosensors, including self-propelled micromotors: their materials, fabrication, and applications. A total of seven papers (five research and two review papers) are included. Different but related topics are covered, from biosensor design to integrated configurations that monitor metabolites in cellular environments. The reviews are devoted to protein-based biosensors and moving biosensors based on self-propelled micromotors. This Special Issue aims to highlight the most recent and promising technologies in micro and nanoscale biosensors, including self-propelled micromotors: their materials, fabrication, and applications. Current progress in nanoscience and nanotechnology has paved the way for the design of many new multi-functional materials for (bio)sensing purposes. Modern biosensors, based on such microscale and nanoscale materials, offer new opportunities in analytical chemistry, healthcare, and many other fields. Indeed, the reduction in size to nanoscale dimensions may result in cheaper, portable, and easier-to-use analytical tools, allowing for real-time analytical measurements, detection in microfluidic systems, and in vivo monitoring applications. Of great interest, self-propelled micromotors represent a new paradigm in the field as novel nanoscale biosensors for the development of \u2018on-the-move\u2019 sensing schemes.This Special Issue compromise five research papers, one feature, and one review article. Two research papers explore different configurations for biosensing purposes. In the first article, Mat\u011bjovsk\u00fd and Pitschmann illustraA second important core of the Special Issue deals with the integration of biosensors in cell environments to monitor metabolites associated with cell events. Kieninger et al. employedThe Special Issue also compromises a feature article devoted to S-layer protein biosensors , which h"} +{"text": "In the Stimulus subsection of the Materials and methods section, there is an incorrect reference in the third sentence.The correct sentence is: Expression stimuli were chosen from the East-Asian Dynamic Facial Expression Stimulus (EADFES) database , which contains stimuli created and verified based on suggested action units related to each emotion .The reference is: Kang J, Ham BJ. Development and Validation of East-Asian Dynamic Facial Expression Stimulus. Proceedings of Current Trends in Biomedical Science. 2015. Seoul, South Korea."} +{"text": "In the original published version of this article, the title and page number of one of the references was incorrectly written. The updated version of the reference is shown below with the changes shown in bold. The authors apologize for this mistake. Both the HTML and PDF versions of the article have been updated to correct these errors.fermented date seed for improving the quality and shelf life of flat bread: Study with univariate and multivariate analyses. Journal of Food Science and Technology, 53(1), 209\u2013220.Najafi, Mohammad B Habibi, Pourfarzad, Amir, Zahedi, Hoda, Ahmadian-Kouchaksaraie, Zahra, & Khodaparast, Mohammad H Haddad. (2016). Development of sourdough"} +{"text": "Scientific Reports 10.1038/s41598-017-01024-8, published online 24 April 2017Correction to: The original version of this Article contained an error in the spelling of H. Jansens, which was incorrectly given as H. Jansen in the Article, and as H. Janssen in the Supplementary Information file.This has now been corrected in the HTML and PDF versions of this Article, and in the Supplementary Information file."} +{"text": "Dear Editor,We report a case of multiple warty dyskeratoma (WD). The patient is a 55-year-old Chinese man who presented with a four-year history of multiple pruritic papules and plaques on the scalp. The number and size of the lesions were gradually increasing. No family history of similar lesions was recorded. Physical examination revealed multiple, discrete, hyperkeratotic papules and plaques, but no vesicles or erosions on his scalp . No otheNone declared.Qiang Zhao: Elaboration and writing of the manuscript.Hongmei Zhou: Obtaining, analyzing and interpreting the data.Songmei Geng: Approval of the final version of the manuscript.None declared."} +{"text": "Nature Communications; 10.1038/s41467-018-06543-0; published online 23 Oct 2018.Correction to: The original version of this Article contained an error in the author affiliations. The affiliation of Alice Chen-Plotkin with the Department of Neurology, Perelman School of Medicine, Philadelphia, PA, 19104 USA was inadvertently omitted. This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "With this trend, many works have already been made to improve the performance of VLC. However, the effectively integration of VLC services into IoT networks has not yet been sufficiently studied. In this paper, we propose a scheme for device management and data transport in IoT networks using VLC. Specifically, we discuss how to manage VLC transmitters and receivers, and to support VLC data transmission in IoT networks. The proposed scheme considers uni-directional VLC transmissions from transmitter to receivers for delivery of location-based VLC data. The backward transmission from VLC receivers will be made by using platform server and aggregation agents in the network. For validation and performance analysis, we implemented the proposed scheme with VLC-capable LED lights and open sources of oneM2M. From the experimental results for Recently, Internet of Things (IoT) services have widely been used to improve our daily life. For example, with the help of IoT services, people can order what they need by pushing a button, or find the location where a fire has occurred by aggregating the measured values from sensors through the Internet.For realizing the IoT, there has been lots of work undertaken on wireless communication technologies such as Bluetooth Low Energy (BLE) and ZigBIn the meantime, some industry areas may be subject to location-critical jobs and/or limited RF communication environments, such as huge factories, airplanes, underground facilities, and so on. Such environments need a specialized communication technology. Visible Light Communication (VLC), which is defined in the IEEE 802.15.7, is one of the communication solutions for meeting these characteristics . VLC comNowadays, LED lights are widely spread in our daily life. A great deal of work has been done on how to improve the performance of LED-based VLC in the physical and MAC layers ,12,13,14This paper is organized as follows. The VLC standard was developed by IEEE 802.15 Task Group 7 in 2011 . The VLCThe IPv6 over Low-power Wireless Personal Area Networks (6LoWPAN) was made to support IP-based communication over the IEEE 802.15.4 Wireless Personal Area Network (WPAN) in the Internet Engineering Task Force (IETF) ,20. The The Constrained Application Protocol (CoAP) was developed for constrained or sensor nodes by the IETF Constrained RESTful Environments (CoRE) Working Group (WG) . CoAP isThe MQ Telemetry Transport (MQTT) is basedoneM2M is a global organization for standard development and efficient deployment of M2M communications systems . Until nAs described previously, much work has been done on how to improve the performance of VLC transmission and on the protocols and platforms for IoT, such as 6LoWPAN, CoAP, MQTT, and oneM2M. However, how to effectively integrate VLC services into IoT networks has not been studied enough. That is, VLC has not been considered as a wireless access medium for IoT. Thus, in this paper, we propose a framework for VLC-based IoT networks and also discuss the operations for device management, data transport and handover in IoT networks using VLC.For VLC-based IoT networks, we consider the following four types of network nodes: Platform Server (PS), Aggregation Agent (AA), VLC Transmitter (VT), and VLC Receiver (VR). There are two possible communication scenarios between VT and VR, as shown in It is noted that most real-world VLC deployments follow the uni-directional model displayed in Platform Server (PS)PS performs overall management for all devices, including AAs, VTs, and VRs, in the VLC-based IoT network through the operations of device initialization, monitoring, and VR handover. PS also controls location-specific VLC data (from VT to VR) and service-specific data (between AA and VR) by using the data transport operation. PS is connected to the Internet.Aggregation Agent (AA)AA manages VT and VR devices locally. It keeps and updates the VT\u2013VR mapping information for its attached VT through the device initialization, monitoring, and VR handover operations. That is, AA can realize the list of VRs that is connected to a specific VT. AA also supports the VR service channel with VR, in which AA exchanges service data with VR. AA is also used to relay location-specific VLC data between PS and VT, and service-specific data between VR and PS.VLC Transmitter (VT)VT controls the LED light. When VT is connected and registered to AA, it transmits location-specific data with VLC frames toward VRs.VLC Receiver (VR)VR is a user or sensor device which acts as a VLC receiver. When VR receives VLC frames with the location-specific data from VT, it will begin the registration with AA. If VR is connected to AA, it may request service data from AA, based on the context of the location-specific data.In In the figure, PS and AA are connected through Ethernet. Connections between AA and VT can be established through Ethernet, WLAN, or WPAN. 6LoWPAN may be used between AA and VT, depending on the underlying link, such as BLE or ZigBee. Each VT is connected to an AA. PS can send location-specific VLC data to each VT via AA. museum service, which consists of a single PS (acting as a museum administrator) and many AAs (an AA may be established at each floor in the museum). It is assumed that each AA is responsible for many VTs, and each VT supports a particular item in the museum . A visitor (user) is equipped with a VR device and moves across VTs to see different items. In this example, PS can provide each VT with location-specific data (an item identifier), and when VR visits the VT, that location-specific information (item identifier) is delivered from VT to VR via the VLC transmission. Based on the item identifier, VR will establish the service channel with AA and download the item (picture) file from the AA. In this way, the location-specific VLC data (possibly initiated by PS and/or AA) is delivered from VT to VR via the uni-directional VLC transmission, and the VR service channel is used between VR and AA for uplink or backward data delivery. This is an example for VLC-based IoT service, and its extension could be possible, depending on the status of VLC deployment and the features of target VLC services.In the meantime, VR receives the VLC data frames from VT. This VLC data frame can contain location-specific data given by PS. Based on this location-specific VLC data, VR establishes the VR service channel with AA by using WPAN or WLAN. To further discuss the use of location-specific VLC data and the VR service channel, let us take an example of the In this section, we describe the VLC-based IoT operations, which are divided into device initialization and monitoring, VLC data transport, and VR handover operations. During device initialization, all of the devices are registered with PS. start and end preamble for distinguishing VLC frames. AA ID and VT ID will be used for VR registration, device monitoring, and VLC data transport. In addition, the information on VR Service Channel can be defined. For example, when WLAN is used between VR and AA, the Basic Service Set Identifier (BSSID) of Access Point (AP) for AA may be included as the information of VR Service Channel. The authorization of VR Service Channel may be added when access control from unauthorized requests is needed. Both fields are related to associating VR Service Channel with AA. A checksum field may be included for checking the validity of the received VLC frame.Device monitoring is used for PS to keep track of the status of the AA, VT, and VR devices in the network. As shown in the figure, AA always keep the AA-VT-VR mapping table for device monitoring. VT and VR may use a timer for periodic reporting . Such a report message contains the identifiers of the concerned VT and VR. On reception of the reports from VT and VR, AA will update its AA-VT-VR mapping table. Then, AA also reports the aggregate status information to PS periodically . It is noted that an appropriate timer value may be configured, depending on the network deployment and the service features to be considered.In the proposed scheme, the VR data is classified into the following two types. One is location-specific data, which is contained in the VLC frames that VT transmits to VRs. The other one is service data, which is delivered through the VR service channel between AA and VR.museum service, the VR device may be used to give detailed information on the concerned exhibition item to the visiting user. In this case, the location-specific data contains the IDs of AA and VT, the channel number of the target item, etc. Please note that such information is specific to a particular VT or item. The VR device might be a smartphone with a mobile guide application for the museum, or a dedicated device that is given by the museum. Based on the location-specific data, VR will try to get more detailed and additional information on the target item from AA, which is served by the service data of VR Service Channel.If VR is a mobile device, it may move around and across different VTs in the network. When a VR moves to another VT, VR performs the handover operation, as described in virtual museum.For validation and evaluation, we implemented the proposed VLC-based IoT scheme by using the oneM2M-based IoT platform and performed testbed experimentation for a simple VLC service, virtual museum service scenario, there are lots of exhibition rooms. Each room has multiple exhibition items, each of which is associated with a VT. virtual museum, in which AA is located at the exhibition room and connected to PS. Multiple VTs are connected to AA through Ethernet in the room. The PS is connected to Internet for museum administrator. A museum visitor (user), who is equipped with VR, will move across different VTs in the room. The VR Service Channel between VR and AA is based on Wi-Fi connection.In the Mobius platform [Mobius server for PS is implemented as Infrastructure Node Common Services Entity (IN-CSE) in oneM2M [nCube-Rosemary for AA is implemented as Middle Node of Common Services Entity (MN-CSE) in oneM2M [nCube-Thyme is used for implementation of VT and VR as Application Dedicated Node of Application Entity (ADN-AE) in oneM2M [Mobius server for PS is installed on a desktop PC with Ubuntu 16.04.1 LTS, and the nCube-Rosemary for AA is installed on a Raspberry Pi with Raspbian OS. VT is also installed on Raspberry Pi with nCube-Thyme and a VLC-capable commercial LED light. Another Raspberry Pi is used for VR with Linux OS, VLC receiver, and nCube-Thyme. PS is equipped with its own DHCPv6 server, which is implemented using the dnsmasq package from the Ubuntu repository. AA also has its own DHCPv6 server, and the functionality of Access Point (AP) is implemented by the hostapd package from the Raspbian repository.For experimentation, we implemented the proposed scheme by using oneM2M-based platform software, called the platform , which in oneM2M , the nCun oneM2M , and nCun oneM2M . The MobField Programmable Gate Array (FPGA)-controlled commercial LED light and a pre-installed firmware for VLC function. An enhanced version of VT will be considered for further study. The VR is attached to the FPGA-controlled Photo Diode (PD) for functionality as a VLC receiver. The VR receives the VLC data through Universal Asynchronous Receiver/Transmitter (UART) communication with the VLC receiver device. The VR service channel between AA and VR is established by using Wi-Fi.Pulse Width Modulation (PWM)-based modulation. The black horizontal lines of VT, as shown in For VLC communication, VT and VR use CNT-AA-SERVICE-DATA-RESP, which is used for receiving service data from PS during data transport operation (see the packets no. 33~40). Then, AA registers to PS by exchanging Common Service Entity (CSE) information . After registration of AA to PS, PS sends the AA Network Configuration message to the container (CNT-AA-CONTROL) of AA (see packets no. 43 and 45).The current version of VT is installed as pre-installed firmware of VLC functions without IP communication. Therefore, the location-specific data is included in the firmware. An enhanced version of VT will be made for further study, which will act as an ADN-AE device of a oneM2M platform. CNT-VR-HEARTBEAT-REQ is for device monitoring, CNT-VR-SERVICE-DATA-RESP is for receiving service data during data transport operation, and CNT-VR-HANDOVER-RESP is for support VR handover.As mentioned earlier, we perform the device monitoring operation periodically. In experimentation, VR is an ADN-AE device, and VT uses uni-directional broadcast VLC. For performance analysis by experimentation, we measured the times required for the VLC data transport and VR handover operations. For each performance metric, 10 experiments are made in the same way, and the min., max., and average values are obtained, as shown in contentInstance of each request. The service data response from PS to AA takes 13 ms, maximum, and service data response forwarding from AA to VR takes 5 ms, maximum. These delays are relatively small, compared to the previous two messages. This is because these two messages do not create contentInstance. The overall delay in receiving the service data is a maximum of 589 ms. In this experiment, we used a tiny piece of data. It may take a longer time if larger data is used.contentInstance of AA. On the other hand, the VR handover ACK from AA to VR takes 9 ms, maximum. The ACK operation does not create contentInstance. The overall handover delay for handover completion is 207 ms, maximum. From the figure, we can also see that the maximum value of handover delay will not exceed 207 ms, because it does not need data.virtual museum service in the experiment is simulated under a testbed environment, in which we performed the experimentations with several real VLC transmitters and receivers. The VLC transmitters used in the experiment are 18 W, white-colored (5700 K), 120 degrees of view angle, 1440 lm flux LED lights. The VLC modulation uses PWM with a 1.6 ms period and 4 m communication range. The PD-based VLC receiver has a 115,200 baud rate through UART.The virtual museum service into real-world environments, we need to consider large numbers of VLC transmitters and receivers, since there are many exhibition rooms and a lot of users in the museum. In this case, the performance may depend on how to effectively configure the platform server and many aggregation agents with VLC transmitters and receivers in the network. In addition, the security issue should be considered, because the VLC frames have no encryption scheme and contain channel information of VR service network. In the deployment of VLC transmitters in real networks, we also need to consider that VLC frames are generated with the same modulation scheme and wavelength, and thus this may cause interference.To extend the virtual museum services. From the experimental results, we see that the VLC data packets can be exchanged within 590 ms, and the handover between VLC transmitters can be completed within around 210 ms in the testbed network.In this paper, we designed and proposed a device management and data transport scheme for IoT networks based on VLC. For validation and performance analysis, we implemented the proposed VLC-based IoT scheme with VLC-capable LED lights and the open-source oneM2M platform. We also conducted testbed experimentation for To the best of our knowledge, this paper is the first proposal for VLC-based IoT. In further works, we plan to implement and investigate the proposed scheme with a protocol stack based on CoAP/UDP/6LoWPAN. We will also consider the bi-directional VLC communication scenario in VLC-based IoT networks. In addition, we will include a comparison of the proposed VLC-based IoT scheme with existing IoT schemes which use other access network technologies, such as Bluetooth and ZigBee, and with other candidate schemes for VLC-based IoT. The testbed experimentation and performance comparisons will be made for a variety of network environments."} +{"text": "This Editorial first introduces the background of the vaccine and drug relations and how biomedical terminologies and ontologies have been used to support their studies. The history of the seven workshops, initially named VDOSME, and then named VDOS, is also summarized and introduced. Then the 7th International Workshop on Vaccine and Drug Ontology Studies (VDOS 2018), held on August 10th, 2018, Corvallis, Oregon, USA, is introduced in detail. These VDOS workshops have greatly supported the development, applications, and discussion of vaccine- and drug-related terminology and drug studies. Drugs and vaccines are critical to public health worldwide. When we discuss drugs, we often mean chemical drugs. Vaccines are typically classified as biological drugs. Both follow similar paths and rules in terms of preclinical research, manufacturing, clinical trials, government approval, and post-licensure usage surveillance and monitoring. However, drugs and vaccines have many differences . For exaIn the time of precision medicine and big data, there has been a huge challenge in organizing, integrating, and analyzing various vaccine and drug related data. The big data can be defined by the typical 4\u2009V model: high volume, high variety, high velocity, and high veracity . Such biNew England Journal of Medicine highlights the critical role of ontologies in standardization, classification, integration, and analysis of various types of knowledge and data associated with diseases, mechanisms, and precision medicine [Over the last decades, we have learned a lot about biomedical ontologies and terminologies and how they can support public health and basic biomedical research. Before the report of the Gene Ontology (GO) , we knowmedicine .The VDOS workshop series have continuously provided a platform for sharing new development and applications of vaccine- and drug-related ontologies, discussing challenges and solutions in the fields, and promoting collaborations among researchers. These workshops usually cover two main areas of topics. One topic is the ontology representation of drugs and vaccines and their associated topics such as adverse event, prescriptions, and molecular mechanisms. The other topic covers various applications of the ontologies in real-world situations such as text mining, machine learning, and software development. Basic and translational research as well as clinical subjects have been widely covered.https://sites.google.com/site/vdosworkshop/VDOS-2018) was held at Corvallis, Oregon, USA, on August 10, 2018. This workshop was part of the ninth International Conference on Biomedical Ontology (ICBO-2018). Overall, VDOS-2018 was another successful VDOS meeting. In this Editorial, we would like to first summarize the results of the previous VDOS meetings and then focus on the introduction of the papers presented in the VDOS-2018 workshop.The 7th International Workshop on Vaccine and Drug Ontology Studies since 2012. The first workshop was named VDOSME-2012, standing for Vaccine and Drug Ontology in the Study of Mechanism and Effect 2012 , a formehttp://easychair.org) to manage our paper submissions and reviewing. All of the papers were peer-reviewed by at least two experts before their acceptance. All of them were orally presented in the workshops and the authors were invited for submitting an extended research article for publication in peer-reviewed journals. Overall, all of these papers except two , using ML- and rule-based approaches. The ML approach employed a deep learning architecture, integrating bi-directional Long Short-Term Memory (Bi-LSTM), Convolutional Neural Network (CNN), and Conditional Random Fields (CRF) for entity recognition. Rule- and dictionary-based approach was implemented on their in-house text mining system, SciMiner [Lastly, i et al. presenteSciMiner , 52, whiOverall, the VDOS-2018 workshop covered six full-length paper representations and offered a platform for sharing the results of vaccine- and drug-related ontology development and applications. A lot of positive feedbacks were provided. We also expect to continue this workshop series in the future and make it an attractive event for more and more ontology and application developers and users."} +{"text": "David Shalloway . Ono andIP2 .A theme in Freed\u2019s research for many years has been the interplay between the gp41 cytoplasmic tail and the MA domain of Gag. His early work had shown that single amino acid mutations in MA block Env incorporation, and that this block can be overcome by deleting the gp41 cytoplasmic tail . Freed\u2019sAn additional focus of Freed\u2019s research since the early 2000s has been the development of HIV-1 maturation inhibitors \u201345. In cResearch in the Freed laboratory has benefited greatly from a number of productive and rewarding collaborations with investigators both within and outside of the NIH, including Schuyler van Engelenburg, Jennifer Lippincott-Schwartz, Juan Bonifacino, James Hurley, Michael Summers, Judith Levin, Eric Barklis, Asim Debnath, Simon Cocklin, Shan-Lu Liu, and Sriram Subramaniam, to name a few , 55\u201374.In 2014, Freed became Deputy Director of the HIV DRP (under then-Director Stephen Hughes) and in 2015 became Director of the HIV DRP, which was renamed the HIV Dynamics and Replication Program. Freed enjoys close scientific interactions with the other DRP PIs\u2014Stephen Hughes, Alan Rein, Vinay Pathak, Wei-Shau Hu, Frank Maldarelli, Mary Kierney, and Alex Compton\u2014and with DRP-associated investigators John Coffin and John Mellors.Viruses. He also serves as an Editor for Journal of Molecular Biology and is on the editorial boards of several virology journals, including Journal of Virology and Retrovirology. Starting in 2018, he is serving as an Associate Editor for Fields Virology. Freed has been an organizer for a number of international virology conferences, including the Cold Spring Harbor Retroviruses Meeting (2004); American Society for Cell Biology Conference on the Cell Biology of HIV-1 and Other Retroviruses (2006); the Keystone Meeting \u201cFrontiers in HIV Pathogenesis, Therapy and Eradication\u201d (2012); the Keystone Meeting \u201cThe Ins and Outs of Viral Infection: Entry, Assembly, Exit and Spread\u201d (2014); two conferences sponsored by Viruses: \u201cAt the Forefront of Virus-Host Interactions and \u201cBreakthroughs in Viral Replication\u201d ; and the International Workshop of the Structure and Function of the gp41 Cytoplasmic Tail . Freed is Co-Chair of the NIH Virology Interest Group and Co-Director of the University of Maryland Virology Program. In addition to ad hoc service on many national and international study sections and grant review panels, he was a member of the NIH AIDS Discovery and Development of Therapeutics (ADDT) Study Section from 2012 to 2017 and served as ADDT Chair from 2015 to 2017. The recognition of which Freed is perhaps most proud is an NCI Mentor of Merit Award for excellence in mentoring.Freed has long been an advocate for virology research and has worked to promote interactions among virologists. Since its launch in 2009, he has served as Editor-in-Chief of the open-access journal When not working, Freed enjoys running, hiking, skiing, biking, backpacking, fly fishing, and music, and spending time with his family."} +{"text": "In the Funding statement, one of the grant numbers is missing. The correct Funding statement is as follows: This work was funded by NIAID/NIH grants R01 AI116813, R21 NS100477, R21 AI140063, and R01 NS106387 to SS and the Chiba-UCSD Center for Mucosal Immunology, Allergy and Vaccine Development. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "In the article by Barratt and Thomas , the secBarratt J (2016) A case study of the nurse practitioner consultation in primary care: communication processes and social interactions. Doctoral thesis/PH.D. Thesis, London: London South Bank University.Barratt J (2005) A case study of styles of patient self-presentation in the nurse practitioner primary health care consultation. Primary Health Care Research and Development6, 327-338.In the second paragraph of the first page, Barratt, 2016 is wrongly listed. The references should read:\u201c.\u201dThe publishers would like to apologise for a further error. In the article, Julian Barratt's job title was incorrectly noted as Head of Community Nursing and Workforce Development. In fact, he is the Head of Community Nursing."} +{"text": "Nature Communications 10.1038/s41467-019-12987-9, published online 8 November 2019.Correction to: The affiliation of Jan Komorowski with the Institute of Computer Science, PAN, Warsaw, Poland, was inadvertently omitted in the original version of this Article. This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "The following information is missing from the Funding section: This work was supported in part by Six Talent Peaks Project of Jiangsu Province, China (2016-WSN-157). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Nature Communications 10.1038/s41467-018-03084-4; published online 22 May 2018Correction to: The original version of this Article omitted a declaration from the Competing Interests statement, which should have included the following: \u2018J.D.B. is a founder and Director of the following: Neochromosome, Inc., the Center of Excellence for Engineering Biology, and CDI Labs, Inc. and serves on the Scientific Advisory Board of the following: Modern Meadow, Inc., Recombinetics, Inc., and Sample6, Inc.\u2019. This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "The emergence and spread of multidrug-resistant tuberculosis (MDR-TB) poses a major threat to the global targets for TB control. In recent years, an evolving science and evidence base for MDR-TB has led to much needed changes in international guidelines promoting the use of newer TB drugs and regimens for MDR-TB, however, there remains a significant implementation gap. Due to the complexity of treating MDR-TB, management of cases is often supported by an expert multidisciplinary team, or clinical expert group. This service is often centralized, and may be delivered through a telemedicine platform. We have implemented a Web-based \u201cstore-and-forward\u201d telemedicine service to optimize MDR-TB patient care in Daru, a remote and resource limited setting in Papua New Guinea (PNG). From April 2016 to February 2019, 237 cases were discussed using the service. This encompassed diagnostic (presumptive) and treatment cases, and more recently, support to the scale up of preventative therapy for latent TB infection. There were 75 cases in which the use of Bedaquiline was discussed or mentioned, with a high frequency of discussions occurring in the initial period (26 cases in the first 12 months), which has appeared to decrease as clinicians gained familiarity with use of the drug (15 cases in the last 12 months). This service has supported high quality clinical care and fostered collaboration between clinicians and technical experts in a shared learning environment. Tuberculosis (TB) is one of the top 10 causes of death globally and is the leading cause of death from a single infectious agent. It is a disease that disproportionally affects low- and middle- income countries, where it remains a major public health issue .The emergence and spread of multidrug-resistant tuberculosis poses a risk to achieving the End TB goals. MDR-TB is a major contributor to mortality and the financial burden of the antimicrobial-resistance threat . MDR-TB In recent years, an evolving science and evidence base for MDR-TB has led to much needed changes in international guidelines promoting the use of newer TB drugs and regimens for MDR-TB, with further changes anticipated in coming years . HoweverTechnical assistance and expertise, including clinical support, either within a country or from international partners plays an important role in optimizing MDR-TB in high burden/resource limited settings . TelemedWe describe the global utility and experience with telemedicine for MDR-TB care and our own experience with the implementation of a Web-based telemedicine platform to support MDR-TB care in a remote setting of Papua New Guinea (PNG). This model has facilitated the delivery of technical assistance to clinicians in the field through a store-and-forward text-based platform suitable for the local context.Telemedicine is a broad term within the domain of digital health, that encompasses a wide scope of practices, all relating to the delivery of health care at a distance . DigitalDue to the complexity of treating MDR-TB, which involves long, complex and expensive regimens and close monitoring for side effects, management of these cases is often supported by an expert multidisciplinary team who provide advice and guidance to the treating clinician. This group has been termed a \u201cTB consilium\u201d in some settings , with orA number of countries, including Belarus, Belgium, France, Mexico, Portugal, and the UK have implemented national level TB consilia , 12. TheThe European Respiratory Society (ERS)/WHO electronic consilium was a suIn many medium and high TB incidence settings, TB consilia have acted as an approval or consensus decision system for accessing new drugs or regimens for MDR TB, specifically relating to the use of the new TB drugs: bedaquiline and delamanid , 16, 17.Finally, consilium can serve an educational role: for clinicians to review how previous similar cases have been managed, and also to have direct access to seek guidance from experienced advisors and mentors. Submitted cases may be rare, unusual or the first experience with a particular regimen or combination of drugs , in whicPapua New Guinea (PNG) is classified as a high burden country for MDR-TB, TB and TB/HIV by the WHO. It has the highest estimated TB incidence rate in the Western Pacific region at 432 per 100,000 people . This leThe Burnet Institute, based in Australia, has been the technical assistance partner in the multi-stakeholder response in Western Province through the RID-TB project (Reducing the Impact of Drug-resistant TB in Western Province) since August 2014. The RID-TB project supports the provincial TB program in program design and evaluation, implementation, clinical care, capacity building and training, health systems strengthening and operational research. The RID-TB project staff include long term advisors (remote/visiting) and field-based TB specialists. Clinical care is provided by medical officers and health workers at Daru General Hospital. A clinical expert group (CEG) (or consilium) was formed to support MDR-TB care and provide advice, primarily on patients to whom the standardized care pathway did not apply e.g., pediatric cases, extra-pulmonary TB, TB-HIV, extensively drug-resistant TB (XDR-TB) and management of adverse effects or complex cases. In particular, there was a need to provide clinical consensus decision for patients initiated on newer TB drugs\u2014bedaquiline and delamanid\u2014which were initially obtained via compassionate access, and for which there was limited in-country experience.Because of the remote location, the CEG initially communicated de-identified case discussions on email and a file server for records. After assessment of various options, the project implemented a telemedicine platform in April 2016: Collegium Telemedicus . The choThe objectives of the consilium/CEG: are to provide clinical support in complex case management, with the additional functions of capacity building, quality assurance, and the formation of collaborations between physicians from different locations .From April 2016 to February 2019, 237 cases were discussed using the platform. This encompassed diagnostic (presumptive) and treatment cases. More recently, support to the scale up of preventative therapy for latent TB infection has been included. Across this time period, there have been 44 different \u201cuser\u201d accounts , and a current pool of five technical experts providing technical support and advice across a range of areas. The median response time from referral for cases was relatively prompt at 17.6 h, facilitating timely decision making. The median number of messages exchanged for each case was 10, reflecting the amount of user engagement and discussions occurring.The program has seen a high turnover of field-based staff, due to the remote and challenging context. As a result, there can be some variation in practices with different clinicians and one key benefit of Collegium Telemedicus that we have noted is for continuity of patient care. With patients often encountering several different clinicians across their \u201cjourney,\u201d this allows transparency in the rationale for previous decision making. As a learning tool, the consilium has also served as a useful resource to explore how similar patients may have been approached in the past e.g., MDR-TB infections in pregnancy.The telemedicine service facilitated the uptake of innovations where technical support was required. The programmatic use of the TB drug Bedaquiline was scaled up in PNG in 2016, procured by the National TB Program, and supported by the global donation program . In thisA number of progress/feedback reports were completed by program participants. Of the five reports that were completed across 2016\u20132018, all respondents felt that the responses they received on Collegium Telemedicus were timely and well-adapted to the local environment. However, in response to whether there would be a benefit to the eventual outcome for the patient, two participants responded \u201cyes,\u201d two responded \u201cperhaps\u201d and one responded \u201cno\u201d. Three of the five respondents felt that there was educational benefit in the process.Although there were a number of concurrent interventions across this time period, we believe that this service has contributed, in part, to the very good outcomes now seen for MDR-TB patients in Daru , 21. TreAn analysis of the perceived benefits and challenges of the telemedicine platform according to the defined objectives, is provided in Implementation of the WHO End TB strategy will require ongoing innovation and the introduction of new treatments and care models for MDR-TB. The WHO guidelines will have a number of significant and ongoing updates as new evidence is generated, in particular for MDR-TB treatment. These will require technical assistance in resource-limited settings, where training and capacity gaps may exist.The RID-TB technical assistance project in Daru, a remote and resource limited setting in PNG, implemented a telemedicine platform to optimize MDR-TB patient care. The telemedicine platform supported high quality clinical care and fostered collaboration between clinicians and technical experts in a shared learning environment. In particular, it bridged the knowledge-delivery gap in supporting the scale-up of innovations such as implementation of bedaquiline for MDR-TB and management of complex cases.Telemedicine is a key intervention to optimize patient care and build local capacity and clinical collaborations in the management of MDR-TB in low resourced settings, particularly in the landscape of new and emerging evidence and evolving standards of care in MDR-TB anticipated in the coming decade.khai.huang@burnet.edu.au.The datasets for this manuscript are not publicly available. Requests to access the datasets should be directed to GH, GP, RW, PdC, and SM: concept. GH and RW: acquisition of data. GH, GP, DO'B, PdC, RW, and SM: drafting of manuscript. GH, GP, MT, SH, PU, DO'B, PdC, SG, RW, and SM: critical review and final approval of manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The Maternal, Infant and Early Childhood Home Visiting (MIECHV) Program is a two-generation approach to supporting healthy families through home visits during pregnancy and early childhood. All states and territories receiving MIECHV funding are encouraged to evaluate their programs. This special issue highlights evaluations from 11 awardees\u2014Arkansas, Florida, Illinois, Iowa, Maryland, Massachusetts, Michigan, New Jersey, Oregon, Pennsylvania, and Tennessee. With the wide expansion of home visiting since the onset of MIECHV, the state-led evaluations contribute to the understanding of replication and scale-up of evidence-based home visiting. The Maternal, Infant and Early Childhood Home visiting (MIECHV) Program is a two-generation approach to supporting healthy families through home visits during pregnancy and early childhood. MIECHV is administered by the Health Resources and Services Administration in collaboration with the Administration for Children and Families. With a $2.7\u00a0billion federal investment since the program launched in 2010, MIECHV has provided over 3.3\u00a0million home visits to at-risk families in all 50 states, Washington DC, and five territories. In 2016, the program served approximately 160,000 parents and children in 893 counties across the United States.Statutory authority for the program from\u00a0the Design Options for Home Visiting Evaluation (DOHVE) project, federal evaluation staff, and federal project officers to design and implement rigorous evaluations.The MIECHV state-led evaluations help answer questions awardees have about their programs while potentially adding to the larger evidence base on home visiting. Specifically, with the wide expansion of home visiting since the onset of MIECHV, the state-led evaluations can contribute to our understanding of replication and scale-up of evidence-based home visiting. Furthermore, by studying the implementation of evidence-based home visiting in new settings and with different cultural and local contexts, the evaluations add to the external validity of the current evidence base. Rigorous impact studies of promising approach models can add to the literature on what works, as do studies of add-ons to traditional evidence-based models. Finally, as the state-led evaluations are required to address questions of interest to programs, themes in their focus contribute to our understanding of implementation challenges and supports.This special issue highlights evaluations from 11 awardees\u2014Arkansas, Florida, Illinois, Iowa, Maryland, Massachusetts, Michigan, New Jersey, Oregon, Pennsylvania, and Tennessee. Initial abstracts were selected based on the rigor and potential relevance of the evaluations for the field. In addition to these criteria, guest editors chose abstracts representing a diversity of topics, methods, geographical distributions, and home visiting models seen in the larger set of MIECHV evaluations. The resulting articles cover four broad topics- understanding and enhancing program quality and fidelity , family engagement , workforce development (Florida and Maryland), and impacts of home visiting, including evidence-based models, promising approaches, and enhancements to models, on key maternal and child health outcomes .State-led evaluations provide valuable information for the MIECHV program and the broader field of home visiting. Thus the Health Resources and Services Administration continues to encourage awardees to pursue questions of programmatic interest. The local evaluations provide an unprecedented body of findings around the implementation and effectiveness of home visiting, complementing ongoing federal and non-federal efforts to learn from and scale up evidence-based programs."} +{"text": "Clinical and Translational Gastroenterology; 10.1038/s41424-018-0045-0; published online 12 September 2018Correction to: In the original version of this article, a footnote was missing. The PDF and HTML versions of the article have now been corrected to include the following:This study was presented as an abstract at the World Congress of Gastroenterology (WCOG) at ACG 2017, Orlando, FL, USA."} +{"text": "Adipose tissue (AT) is strongly associated with development and progression of immune disorders through adipokines secretion, such as adiponectin. This protein has beneficial energetic properties and is involved in inflammation and immunity processes. Three oligomers of circulating adiponectin with different molecular weight are described: High (HMW), Medium (MMW), and Low (LMW). The HMW is the most biologically active oligomers. On binding to its receptors AdipoR1, AdipoR2, and T-cadherin, adiponectin acts on both innate and acquired immunity. The suppression of NF-\u03baB activation and pro-inflammatory cytokine expression in macrophages is mediated by AdipoR1. AdipoR2 mediates polarization of anti-inflammatory M2 macrophages T-cadherin is essential for the M2 macrophage proliferation. Furthermore, adiponectin reduces T cells responsiveness and B cells lymphopoiesis. The immune system is very sensitive to environmental changes and it is not only interconnected with AT but also with the central nervous system (CNS). Cytokines, which are mediators of the immune system, exercise control over mediators of the CNS. Microglia, which are immunity cells belonging to the macrophage family, are present within the CNS. The nervous system is also involved in immunity through the production of neuropeptides such as orexin-A/hypocretin-1. This neuropeptide is involved in metabolic disorders, inflammation and in the immune response. The relationship between adipokines, immunity, and the nervous system is validated by both the role of orexin-A on fat, food intake, and energy expenditure, as well as by role of adiponectin on the CNS. In this review, we focused on the functions of adiponectin and orexin-A as a potential immunity link between AT and CNS. Adipose tissue (AT) is a multifunctional organ involved in many physiological and metabolic processes. Its functions are both as a site for energy storage and as an endocrine organ, fundamentally composed of adipocytes but populated also by a number of immune cells such as T lymphocytes and macrophages . As a reThe AT is the main deposition of triglycerides in mammals, including man. One of the main functions of adipocytes is the synthesis and release of triglycerides. The number of cells and the dimension of adipose mass depend on the nutrition state of the individual. Excessive caloric intake (obesity) results in an increase in adipose mass, while chronic nutritional deficiencies result in the opposite outcome. It is well known that AT is constituted not only by adipocytes but by a plethora of different immune cells, such B cells, T cells, as macrophages, and dendritic cells that produce anti-inflammatory cytokines . In a chFigure 1). It increases the sensitivity to insulin, stimulates glucose, and lipid metabolism, and provides a protective factor for cardiovascular diseases (in vitro study on chondrocytes suggested that adiponectin promotes rheumatoid arthritis (Adiponectin is the most abundant adipokine produced by AT. This protein can be found in three different molecular weight: high (HMW), medium (MMW), and low molecular weight (LMW). The HMW adiponectin oligomers are the most biologically active. Adiponectin acts by binding two membrane specific receptors located in numerous organs and tissues: AdipoR1 and AdipoR2. These receptors belong to the seven-transmembrane receptor family coupled with G proteins and possess the N-terminal domain within the cell and the C-terminal outside. AdipoR1 and AdipoR2 differ in the specificity of binding to the adiponectin. AdipoR1 is activated both by the full-length as well as by the globular form o adiponectin. Instead, AdipoR2 has more affinity for the full-length form. A third receptor of the adiponectin is T-cadherin. The latter is a calcium-dependent adherence molecule that presents the classical cadherin structure, even if it does not have a cytoplasmic or transmembrane domain. T-cadherin is widespread ubiquitous, with high expression in the cardiovascular system and low levels in the muscles. T-cadherin recognizes only the MMW and HMW oligomers . Throughdiseases . Interesdiseases . On the diseases . As repodiseases . Adiponediseases . A pivotdiseases . Moreovediseases . On the diseases . The litdiseases . Hietaharthritis . On the rthritis .These studies suggest a controversial role for adiponectin plasma levels and inflammation. Summarily, it could be suggested pro-inflammatory effects in classic chronic inflammatory/autoimmune disease and an anti-inflammatory action in obesity patient.The nervous system plays important functions monitoring and coordinating organ function and responding to changes in the external environment. This system can be divided into two parts, the CNS, and the peripheral nervous system, and includes the brain, spinal cord, and a complex network of neurons. The CNS has involved in different processes through the production of neuropeptides such as orexin-A/hypocretin-1 . RecentlIn addition to the mechanism listed above, the nervous system is also involved in immunity through the production of neuropeptides such as orexin-A/hypocretin-1 .The orexin-A is a neuropeptide produced in the lateral hypothalamus by definite neurons . In a paRecently, another role of the orexin-A in inflammation has been recognized. Expression of orexin-A in rats exposed to ischemia/reperfusion was reported to be very high in the stomach, lung, and kidney . Since iin vivo study\u201d a direct link between adiponectin and orexin-A. Particularly, their findings suggested that orexin-A stimulates glucose uptake in adipocytes, increasing lipogenesis, inhibiting lipolysis, and stimulating the secretion of adiponectin (Here, we reported that recent data on adiponectin and orexin-A provide bases for a potential link between AT and CNS. Both the nervous system and AT actively communicate to the immune system through the production of chemical mediators. In particular, adiponectin and orexin-A are two hormones that play crucial roles in the immune system. Adiponectin is actively involved in the immune response by regulating both macrophage proliferation and polarization acts on monocytes and reduces T cells responsiveness, and B cells lymphopoiesis. On the other hand, orexin A is involved in both inflammatory and immune processes. In particular, this peptide is believed to be responsible for the autoantibodies induced damage to neural cells. ponectin . Moreoveponectin . This coThe current knowledge strongly encourages further research on the potential functional mechanisms through which adiponectin and orexin-A exert their different actions in the immune system. Further studies are needed to clarify the molecular mechanism and the potential functional interplay between these two mediators.RP, EN, AM, AD, and GM conceived the study and participated in its design. RP, MLM, VM, GC, AV, AD, and GM contributed to the conception and design. OS, EP, CZ, DP, and GM wrote the manuscript. MM, IC, VM, and RP drafted the article and revised it critically for important intellectual content. RP, EN, GM, and AD performed the final approval of the version to be published. All the authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "The are errors in the Funding statement. The correct Funding statement is as follows: This study was supported by USDA-NIFA 1890 Evans Allen Award, Grant Number NI181445XXXXG007 and NASA, Grant Number 80NSSC17K0653. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."} +{"text": "Knowledge of end-of-life (EOL) care and being able to make the appropriate decisions for patients who are dying is important for nursing students, who will soon play a critical role in EOL decision-making among patients and their caregivers. Accordingly, the purpose of this study was to examine the level of knowledge of EOL care, life-sustaining treatment, and advance directives among nursing students in South Korea. This cross-sectional descriptive study was conducted from December 2017 to February 2018. Data were collected from 220 undergraduate nursing students and analyzed using descriptive statistics, t-tests, one-way ANOVA, and a post hoc test with the SPSS 19.0 program. The score for knowledge of EOL care was 7.8 out of 11 points, for knowledge of life-sustaining treatment was 4.6 out of 6 points, and for knowledge of advance directives was 7.0 out of 9 points. There were significant differences in knowledge of EOL care scores by year of study, experiences in clinical practicum education, and experiences of caring for dying patients. Knowledge of life-sustaining treatment significantly differed by year of study, experiences in clinical practicum education, experiences of caring for and observing dying patients during clinical practicum education, and perceived self-rated health. There were significant differences in knowledge of advance directive scores by year of study, satisfaction with nursing major, experiences in clinical practicum education, and experiences of caring for and observing dying patients during clinical practicum education. Further studies should develop educational intervention programs that improve knowledge of EOL care, life-sustaining treatment, and advance directives."} +{"text": "Correction to:Cell Death and Disease10.1038/s41419-019-1847-z published online 16 August 2019In the published version of this paper, an institutional co-affiliation of author Dr. Dagmar Quandt was inadvertently omitted. The following institution should also have been affiliated to the study: Institute of Anatomy and Cell Biology, Faculty of Medicine, Martin Luther University Halle-Wittenberg, Halle , Germany. This has been corrected in the PDF and HTML versions."} +{"text": "In cancer research, cell-based assays are used to assess cell migration and invasion. The major bottleneck is the lack of automated tools to visualize and analyse the large amounts of biological dose-response data produced. To address this challenge, we have developed an automated and free software package for dose-response analyses, DoRes, which is released as an add-on of the freely available and open-source tool CellMissy, dedicated to the management and analysis of cell migration data. DoRes implements non-linear curve fitting functionality into a robust, user-friendly and flexible software package with the possibility of importing a tabular file or starting from a cell migration experiment. We demonstrate the ability of the software by analysing public dose-response data and a typical cell migration experiment, and show that the extracted dose-response parameters and the calculated statistical values are consistently comparable to those of the widely used, commercial software GraphPad Prism.https://github.com/compomics/cellmissy/.The software here presented is a new module in CellMissy, an open-source and cross-platform package dedicated to the management, storage and analysis of cell migration data. The new module is written in Java, and inherits the cross-platform support from CellMissy. Source code and binaries are freely available under the Apache2 open-source licence at Bioinformatics online. In vitro cell-based assays are often used to test how candidate components or treatments influence certain cellular functions. Effects on cell proliferation, adhesion, migration, invasion, cell apoptosis, morphology changes and more can be measured . A drawback of GraphPad is that it is commercial and, despite an extensive user guide and support page, the software remains a black box to the user as to the code and algorithms employed to analyse data. Moreover, GraphPad lacks a direct connection to a data storage system, requiring the user to manually annotate and import the relevant data. Here, we therefore introduce DoRes, a free, open-source and automated module for dose-response analysis in the CellMissy software framework. CellMissy is an open-source, cross-platform data management and analysis system for cell migration data that simplifies and automates data management, storage, quality control and analysis can be applied to the curve fitting. Moreover, DoRes bundles all results in a detailed analysis report, which provides a general overview of the experiment, all plots and statistics and information on any applied normalization (see https://cran.r-project.org/web/packages/drc/drc.pdf) data sets see . This chtion see . To demopdf) see . As showpdf) see . ImportaDue to the need to preformat and import the relevant data by hand, the current use of generic, and often commercial dose-response tools is suboptimal (see Supplementary InformationClick here for additional data file."} +{"text": "Nature Communications 10.1038/s41467-019-09319-2, published online 25 March 2019.Correction to: The original version of this Article omitted the following from the Acknowledgements:Gabriel Arellano was instrumental in collecting post-Mar\u00eda Hurricane data, as part of the Next Generation Ecosystem Experiments-Tropics, funded by the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research.This has now been corrected in both the PDF and HTML versions of the Article."} +{"text": "Xiao-peng Ma, Bei Xia, Lie-zhen Hu, Shu-min Fan and Dong Xiao were not included as authors in the published article. The corrected Author Contributions statement and Acknowledgments appear below. The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.XL wrote the paper and conceived and conducted the study. BX designed and improved the procedure of the technique and ensured its quality and safety as the ultrasonography department lead. H-kY wrote the paper and carried out the analysis. L-zH and S-mF implemented quality control of the procedure. DX provided valuable support in the therapeutic process. L-xG and J-kC contributed to data collection and management. Z-bW contributed to the discussion, supervised, and reviewed the text. X-pM promoted the clinical application of the technique. All authors read and approved the manuscript."} +{"text": "To compare individual case safety reports (ICSR) rates and characteristics between Croatia, Serbia, Montenegro, and Bosnia and Herzegovina (B&H).This retrospective pharmacoepidemiological study used the data from ICSR received by the Agency for Medicines and Medical Devices in B&H in 2011-2016. The number, characteristics, and sources of reports, suspected drugs, and patient characteristics were analyzed. The results were compared with the publicly available data from Croatia, Serbia, and Montenegro.The number of reported adverse drug reactions per one million of inhabitants was lowest in B&H and highest in Croatia. There were significant differences in reporter characteristics, sources of reports, and the percentage of missing data in ICSR, while the Anatomical Therapeutic Chemical product classes, patient\u2019s sex, and adverse drug reaction System Organ Classes were similar.Despite the historical and geographical vicinity of B&H and its neighboring countries, there were significant differences in indicators of pharmacovigilance development. Preclinical and clinical investigations of a medicinal product cannot provide complete information on its safety. Therefore, post-marketing evaluation is needed to collect data on adverse effects and contribute to the safer use of medicinal product. To answer the questions related to adverse drug reactions (ADR), every country has to have a well-organized pharmacovigilance system . The devBosnia and Herzegovina (B&H) and its neighboring countries \u2013 Croatia, Serbia, and Montenegro were part of Yugoslavia until 1991. Yugoslavia had a well-organized pharmacovigilance system and participated in the World Health Organization (WHO) Programme for International Drug Monitoring since 1974 . AlthougThis retrospective pharmacoepidemiological study analyzed ICSR received by the Agency for Medicines and Medical Devices in B&H (ALMBIH) in the period 2011-2016. The results were compared with the publicly available results from Croatia, Serbia, and Montenegro -5. We alICSR received by the ALMBIH by mail, email, fax, or through online reporting system for marketing authorization holders (MAH) were used as data source for B&H. For Croatia, Serbia, and Montenegro, data were obtained from publicly available yearly reports. For Croatia and Montenegro, the reports were available for the period 2011-2016. For Serbia, the reports were available for the period 2011-2015 and did not contain all relevant data. Additional data could not be obtained from Medicines and Medical Devices Agency of Serbia (ALIMS) directly upon request. Thus, for comparison of the development level of pharmacovigilance systems, available data from Serbia were used. Data on B&H population were obtained from the Agency for Statistics of B&H . The number, characteristics, and sources of ICSR, suspected drugs, ADRs, and patient characteristics were analyzed quantitatively. The first level of ATC classification was used to characterize suspected drugs in ICSR. ADRs were coded according to the Medical Dictionary for Regulatory Activities System Organ Class (SOC) classification. One ICSR represented one report for a specific patient and can contain more than one ADR and more than one suspected drug. Therefore, the number of ADRs and suspected drugs was higher than the total number of ICSR. Previously published data from Croatia .The normality of data was evaluated by Shapiro-Wilks test. The number of ICSR per one million of inhabitants, percentage of pharmacists, physicians, health care professionals, MAH, and patient reporters and percentage of female patients were normally distributed. The significance of the relationship between analyzed parameters was assessed using type I analysis of variance (ANOVA) test. Since a part of data for Serbia was missing, Games-Howell The number of ICSR was lowest in B&H and highest in Croatia. Serbia and Montenegro had similar number of reports, which was lower than in Croatia and higher than in B&H .P=0.027, P=0.026, and P=0.003, respectively), while no significant difference was found between B&H, Serbia, and Montenegro (P\u2009=\u20090.004) (Percentage of pharmacist reporters in Croatia was significantly higher than in B&H and Montenegro (ctively) . Data on=\u20090.004) . There wATC classes were similar in all countries . Most paThere were significant differences between the four countries in the ICSR number, sources, and percentages of missing data in ICSR. The data for dominant ATC classes of products, patients' sex, and ADR SOC classifications were similar.The number of ICSR in a country directly indicates the pharmacovigilance system development. According to WHO, each national pharmacovigilance center should send at least 200 ICSR per million inhabitants yearly . B&H hadCroatia, on the other hand, had the highest number of ICSR, which was constantly growing during the analyzed period. Croatia has more than 40-year-long pharmacovigilance tradition, as the National Pharmacovigilance Center of Yugoslavia was located in Zagreb . The AgeSerbia and Montenegro had similar number of reports, which was lower than in Croatia and higher than in B&H. In Montenegro, the Agency for Medicines and Medical Devices of Montenegro (CALIMS) plays a very important role in the pharmacovigilance promotion and education. A reporting system is constantly being upgraded, with the main focus on online options. Online forms are well-developed and have many mandatory fields. CALIMS developed projects and collaborations for pharmacovigilance system improvement . In SerbTypes of reporters are among the main indicators of the awareness of ADRs reporting importance. Considerable level of patient reporting was only present in Croatia. This might be explained by HALMED activities in pharmacovigilance education of patients. Croatia was the first country in the world to launch a WHO online reporting system for patients in 2012 . PharmacQuality of reports can be evaluated through the percentage of unknown data. High percentage of ICSR with unknown sex and age data from B&H for 2011 and 2012 indicates the low quality of reports. The decrease in the percentage of incomplete reports after 2012 could be attributed to education of health care professionals performed through ALMBIH website and lectures at relevant conferences.There is a number of studies on the difference between pharmacovigilance regulations and development between countries -22, but A limitation of our study is a lack of some relevant data (for Serbia for 2016 and partly for previous years). Moreover, the analyzed period was short and the number of analyzed countries small. To assess the factors affecting the pharmacovigilance system development we should analyze a larger number of countries during a longer period.Since pharmacovigilance plays a crucial role in safer drugs use, all countries should put in effort to develop well organized systems, producing enough relevant data to optimize risk-benefit analysis in therapy. Some of the activities for countries with less developed pharmacovigilance systems should be continuous education of health care professionals, leading to their active participation in pharmacovigilance; more intensive pharmacists' involvement in ADR reporting; regulation and improvement of patients\u2019 reporting; implementation of online and other adequate tools and ADR reporting systems accessible to all potential reporters; and cooperation and exchange of information between countries."} +{"text": "Journal of Gerontology: Social Sciences aims to publish the highest quality social scientific research on aging and the life course in the U.S. and worldwide. The disciplinary scope is broad, encompassing scholarship from demography, economics, psychology, public health, and sociology. A key substantive focus is identifying the social, economic, and cultural contexts that shape aging experiences worldwide. In the coming decade, social gerontology research is poised to present many opportunities for cross-national and cross-cultural scholarship \u2013 driven in part by the proliferation of large parallel data sets from many nations in Europe, Latin America, and Asia. I will discuss the role that peer-reviewed cross-national scholarship can play in disseminating knowledge that informs gerontological research, policy, and practice internationally. I will also identify under-researched areas that will be of great interest to scholars in the coming decade, including LGBT older adults, aging in the Global South, reconfigured families, and centenarians."} +{"text": "Osteoarthritis (OA) is one of the most common musculoskeletal conditions worldwide, affecting the functional abilities of millions of people. Arthroplasty is recommended as a successful treatment option for late-stage OA. However, in South Africa there are extensive waiting lists for OA-related arthroplasty in government hospitals. This has negative consequences for patients having to cope for long periods of time with chronic pain and its impact. Alternative treatment methods in the form of physiotherapy-led exercise and education programmes focusing on pain, disability, self-efficacy, physical function and health-related quality of life have had good impact in populations elsewhere.To develop an exercise and education intervention based on the current literature and by doing a field survey in a South African population.A combined educational approach, with a strong focus on the physical aspects of exercise in particular, was adopted for the intervention in order to improve function and manage the disability associated with OA.This paper reports on the process and development of an intervention for use in South Africans with late-stage OA awaiting arthroplasty. Worldwide, the prevalence of musculoskeletal conditions is high Brooks . One of et al. In South Africa, the prevalence of OA is similarly high guidelines .et al. musculoskeletal conditions, arthritis, osteoarthritis, chronic pain, health related quality of life, disability, function, self-efficacy, South Africa, ICF, epidemiology, intervention programs, self-management programs. In addition, chronic pain management programmes in use at public health institutions were reviewed and their recommendations for the management of OA guidelines state that local muscle strengthening exercises and general aerobic fitness should be part of core treatments for people with OA, irrespective of age, co-morbidity, pain severity or disability (McAlindon et al. (et al. et al. The exercise components for the intervention designed for the South African population combined aerobic, strengthening and flexibility activities and was largely based on the ESCAPE knee pain programme by Hurley et al. , 2009 (Tet al. . The con. et al. . Furtheret al. et al. Late-stage OA predominantly presents as chronic pain with several psychosocial factors influenced by/influencing the pain experience (Gatchel Diezeman reports et al. et al. While the above evidence for relaxation training is mainly based on populations with chronic low back pain, headaches and fibromyalgia, recent literature highlighting the central nervous system changes present in late-stage OA supports the application of the approaches in this population (Lluch Social isolation is a known problem for patients with chronic illnesses (Chan & Chan et al. et al. One of the biggest obstacles in modern healthcare practice is adherence to guidelines and despite evidence-founded recommendations, their practice is often not followed (Walsh & Hurley et al. et al. et al. CDSMPs commonly range from 6\u201310 weeks in length (Foster https://open.uct.ac.za/handle/11427/12697. The effectiveness of the intervention has been tested in a randomised trial, the results of which are reported elsewhere.This paper illustrates the process followed to develop an evidence-based 6-week physiotherapist-led exercise and intervention for South African patients with late-stage OA of the hip or knee. The \u2018Living with Osteoarthritis\u2019 bio-psychosocial intervention was specifically developed for the management of pain, function, disability and health-related quality of life in patients waiting for arthroplasty of the hip or knee because of OA. The workbook can be accessed at:"} +{"text": "Journal of Molecular Cell Biology on RNA biology, combining a series of reviews and research articles that reflect most of the current trends and topics emerging from recent Chinese\u2013German interactions in RNA research.NA biology has developed in recent years to a new emerging field, with new surprises and insights ranging from basic mechanisms to topics of clinical interest and opening up biotechnological applications in molecular medicine and novel therapeutic options. Here, we edit this Special Issue of The Sino-German Symposium on RNA Biology has been held since 2016, each year concentrating around a special theme of RNA research. The symposium is funded by the Sino-German Center for Research Promotion, a joint venture between the Deutsche Forschungsgemeinschaft (DFG) and the National Natural Science Foundation of China (NSFC), which supports scientific collaborations between China and Germany, in particular workshop-type meetings with ~40 scientists from both countries, all presenting their latest work, discussing ongoing projects, and initiating new co-operations. Renowned scientists from other countries were also invited, such as Prof. Xiangdong Fu from UCSD, USA. Albrecht Bindereif and Zefeng Wang co-organized the first two meetings, which were held in the castle of Rauischholzhausen focusing on the role of RNA in human disease and at the Chinese Academy of Sciences (CAS) campus in Shanghai on RNA-related complex structures and novel mechanisms, respectively. The 3rd meeting co-organized by Markus Landthaler from the Max-Delbr\u00fcck-Center for Molecular Medicine (MDC) in Berlin and Wei Chen from the Southern University of Science and Technology (SUSTech) in Shenzhen was held last year in Berlin and emphasized emerging technologies and new trends in RNA research. The 4th meeting will soon take place in Shenzhen in November 2019, continuing this short but successful history.These Sino-German RNA meetings also rely on a core of long-standing interactions, collaborations, and personal friendships, some extending over the last two decades. For instance, Wei Chen spent many years in Berlin , with a long history of successful German\u2013Chinese collaborations; Jingyi Hui once worked in Albrecht Bindereif\u2019s group in Berlin and Giessen before moving back to Shanghai; and Zefeng Wang co-organized the Otto-Warburg Summer Schools alternating between Berlin and Shanghai and recently became the Mercator Fellow of a DFG-funded Research Training Group coordinated by Albrecht Bindereif. In addition, new collaborations were initiated during these meetings, reflecting the need for an interaction forum for German and Chinese scientists in RNA research. We are very grateful to the Sino-German Center for Research Promotion (Beijing) for continuous support and funding to make this possible.6A RNA modification in plants , with the following topics: RNA structural biology , aging an plants , new insn plants , regulatn plants , generalmechanisms of auto-regulation by RNA-binding proteins , as well"} +{"text": "It is well recognized that national academies of science have a long tradition of engaging widely to strengthen the evidence base to underpin the delivery of enhanced food and nutrition security at regional and national levels. The Association of Academies and Societies of Sciences in Asia (AASSA) has recently produced a report for audiences of the Asia-Pacific regions as a contribution to a project worldwide initiated by the InterAcademy Partnership (IAP), the global network of science academies. The IAP work brings together regional perspectives in parallel from Africa, Asia, the Americas and Europe on the opportunities for the science-policy interface, identifying how research can contribute to resolving challenges for agriculture, food systems and nutrition, of which livestock production constitutes a very important component.AASSA report on Food and Nutrition Security and Agriculture (FNSA) focusses on FNS in the Asia/Oceania region, but given the size and importance of the region and the considerable inter-connectedness of matters influencing food production, food consumption, nutrition and human well-being, many of the observations and recommendations made are global in nature. The target outcome of its deliberations is future FNS, that is, access for everybody to a diverse healthy diet that is underpinned by a production/distribution/consumption system that is sustainable, environmentally, socially and culturally. It is recognized that there are many facets to FNS, and a systems approach to analysis is recommended, where the complex interactions between scientific and technical , economic, political, social and cultural dimensions are considered together. Having said this, in preparing this report the working group, and reflecting its expertise and mandate, has chosen to focus on the crucial role to be played by S&T (both R&D and education) in securing future FNS. Increasing pressures from population growth, urbanization, land availability, resource and water availability, pollution, global climate change, biodiversity loss conspire to make FNS a formidable near-term challenge.This The report identified key S &T areas having universal and prioritized application across the region as followings: (1) Genomic-based approaches to plant and animal breeding. (2) Big-data capture and analysis, precision agriculture, robotics. (3) Food technology innovations in harvest, processing and storage to reduce food wastage. (4) Sustainable farming practices for land and water use, that address wider issues such as biodiversity and climate. (5) Aquaculture production and integrated farm production systems.The Global Report from IAP, analysis and synthesis of four regional reports from Asia, Africa, America and Europe, addresses recommendations for international scientific priorities as follow:Developing sustainable food and nutrition systems, taking a systems perspective to deliver health and well-being, linked to transformation towards the circular economy and bio-economy.Emphasising transformation to a healthy diet and good nutrition.Understanding food production and utilisation issues, covering considerations of efficiency, sustainability, climate risks and diversity of resources.Capitalising on opportunities coming within range in the biosciences and other rapidly advancing sciencesAddressing the food-energy-nutrients-water-health nexus, recognising that boundaries are blurred.Promoting activity at the science-policy interfaces and reconciling policy disconnects. involving them in strategic decisions about planning research.Consolidating and coordinating international science advisory mechanisms.http://www.interacademies.org/37646/Food-and-Nutrition-Security-and-Agriculture.It is a must to readers who are looking for an up-to-date and comprehensive study with scientific and technological perspective on challenges and opportunities of food and nutrition security and agriculture both in Asian and global level. These reports can be enjoyed through websites: Finally, the reviewer would like to congratulate IAP and AASSA for producing well-integrated and resourceful reports with STI perspective on FNSA of both Asia and Global status."} +{"text": "In the version of this article initially published, Candice A. Quarmby\u2019s affiliation was incorrectly mentioned as Disciplines of Audiology & Speech-Language Therapy, University of KwaZulu-Natal, South Africa. Her correct affiliation is Discipline of Speech-Language Pathology, University of KwaZulu-Natal, South Africa. The error has been corrected in the PDF version of the article. The author apologises for any inconvenience that this omission may have caused."} +{"text": "From her perspective as director of ApparentPlan, a nonprofit care agency to assist low income consumer of LTC, and as co-founder and director of the MN Long-Term Care Think Tank, an advocacy organization. Ms. Keibler will reflect on these findings and next steps."} +{"text": "The NIA\u2019s Butler-Williams Scholars Program and GSA\u2019s Emerging Scholars and Professional Organization are united in providing career development opportunities for early career scholars in a manner that promotes leadership, diversity, and inclusivity. This provides a foundation to develop a network of next generation of scientists, clinicians, and policy makers capable of shaping health in aging. Among the chief concerns of our aging population are disparities in health associated with race/ethnicity, experience, environment, access to health care, and sociocultural and socioeconomic factors. GSA\u2019s early career professionals and alumni of the prestigious NIA Butler-Williams Scholars Program have tackled these issues directly and the scientific scholarship that results is astounding in its breadth and depth. Dr. Vicki Johnson-Lawrence, Ph.D. (Butler-Williams class of 2014), will present on the interacting effects of education and race/ethnicity on multi-morbidity, highlighting lessons learned from the National Health Interview Study. Dr. Lauren Parker (Butler-Williams class of 2018) will review efforts to develop culturally competent content for recruitment of Hispanic and black/African American persons to NIA-supported dementia-caregiving studies. Dr. Ryon Cobb (Butler-Williams class of 2016) will discuss the impact of race/ethnicity on kidney function among older adults, with evidence from the Health and Retirement Study. The final speaker, Dr. Ana Qui\u00f1ones (Butler-Williams class of 2012), will present on longitudinal tracking of multi-morbidity in racially/ethnically diverse older adults. In sum, the featured talks by rising stars in aging research deepen our understanding of the influence of race, ethnicity, and experience on health and chronic disease in diverse aging populations."} +{"text": "Al-Shahi Salman R, Frantzias J, Lee RJ, et al. Absolute risk and predictors of the growth of acute spontaneous intracerebral haemorrhage: a systematic review and meta-analysis of individual patient data. Lancet Neurol 17: 885\u2013942018; \u2014The affiliation of Department of Neurology, University of Erlangen-Nuremberg, Erlangen, Germany has been added for authors Dimitre Staykov and Bastian Volbers. This correction has been made to the online version as of Sept 19, 2018."} +{"text": "Extra nodal natural killer/T-cell lymphoma (ENKTL), nasal type is a rare and highly aggressive type of non-Hodgkin's lymphoma (NHL) commonly presented in the nasal cavity or lymphatic system. However, the common causes of mortality in ENKTL remain unclear. We conducted a retrospective population-based cohort study to elucidate the different causes of mortality in ENKTL and illustrate the main causal and associated risk factors leading to death.The study included patients diagnosed with ENKTL from 1987 to 2014 in the Surveillance, Epidemiology, and End Results (SEER) program. Univariate survival analysis was conducted using Kaplan-Meier analysis, and multivariate analyses were performed using Cox proportional hazards regression model. Competing-risks regression model was applied to estimate specific risks associated with mortality.The analysis demonstrated increased mortality in males and patients diagnosed at older age and higher disease stage. NHL was the most common cause of mortality in patients with ENKTL, accounting for 74.13% of deaths in the cohort, followed by other malignant cancers, heart diseases, and infection. However, NHL-specific death events were fewer in patients diagnosed with advanced disease stage compared with incidences of death by other causes such as disease of heart and infections. Significant difference was seen between patients diagnosed earlier than 2000, who showed a higher probability of dying from NHL, and those diagnosed later, who showed propensity to die from other malignant tumors and infection. No differences were found when comparing sex or age at diagnosis.The most common cause of mortality in cases with ENKTL-NT is NHL. The female sex, diagnosis at young age and early stage are associated with improved prognosis. Further, the classification of Ann Arbor stage and year of diagnosis can provide references of specific causes of death, which might help decrease the mortality rate. Extra nodal natural killer/T-cell lymphoma, nasal type (ENKTL-NT) is a comparatively rare and highly invasive type of non-Hodgkin's lymphoma (NHL) that originates from NK cells or T cells and is commonly presented in the nasal cavity or lymphatic system.\u20136 The ovBluhm reported that the causes of mortality in cases diagnosed with NHL were NHL, leukemia, other malignant tumors, cardiovascular diseases and infections. Further,www.seer.cancer.gov/seerstat). SEER is supported by the Surveillance Research Program (SRP) in NCI's Division of Cancer Control and Population Sciences (DCCPS) (www.seer.cancer.gov).Information regarding the participants was downloaded from the Surveillance, Epidemiology, and End Results (SEER) Program , which wCases were selected based on the classification of lymphoid neoplasms published by International Lymphoma Epidemiology Consortium (InterLymph) Pathology Working Group, according to the WHO classification of lymphoid neoplasms and the International Classification of Diseases-Oncology, Third Edition (ICD-O-3). Cases diagnosed with ENKTL-NT with adequate available information were selected for this study. No restricting criteria were added for age, sex, and race. A total of 163 patients participated in the program from diagnosis till the end of a 5-year follow-up or death, whichever occurred first. However, since we aimed to analyze the cause of death, all patients who survived past the follow-up period were excluded from our study. In addition, all patients with death certificate only or autopsy only were excluded, as were patients whose survival time could not be ascertained. The patients included were those diagnosed with ENKTL as the first primary tumor. The study focused on analyzing the primary cause leading to death. Finally, based on our inclusion and exclusion criteria, 107 patients were analyzed for the cause of death and divided into two groups: NHL-specific mortality and mortality by other causes. The latter was further classified in four sub-groups: (1) other malignant tumors, (2) diseases of heart, (3) infections, and (4) other causes.Variables analyzed included sex , calendar year of diagnosis , race , Ann Arbor Stage of the disease (stage I and II or stage III and IV) and age at diagnosis (< 60 or \u2265 60).Distributions of survival time were assessed grouped by basic information and Ann Arbor stage of disease in patients with ENKTL, nasal type. The ICD-10 codes for different causes of death were used in the study to standardize the analysis. The overall survival (OS) was defined as the date from diagnosis to the date of death or the end of the follow-up.https://ecomfe.github.io/echarts-doc/public/en/index.html). All statistical analyses were performed using IBM SPSS Statistics for Windows, version 21.Descriptive statistics, including median and confidence intervals (CI), were provided for continuous variables. Frequencies and percentages were used to summarize categorical variables. For comparisons of OS between subgroups classified by sex, Ann Arbor Stage, race and age at diagnosis of patients, survival curves were drawn using the Kaplan-Meier method . Univariate analyses were performed by the log-rank test while multivariate analyses were performed using the Cox proportional hazards regression model. The pie nest chart was drawn using Echarts to analyze the competing events. When calculating NHL-specific mortality, death by NHL was the event of interest and other-causes mortality was the competing event, and vice versa. Similarly, when considering the sub-group \u2018other causes,\u2019 the four groups were competing events against each other. The analysis of each cause was further grouped by sex, Ann Arbor Stage, calendar year of diagnosis and age at diagnosis of patients. The cause-specific sub-hazard ratios (SHRs) and cumulative incidence function (CIF) were calculated to demonstrate competing risks. We calculated the CIFs for stage, age group, and calendar year of diagnosis. All statistically significant differences were defined as p < 0.05.This study included 163 participants with adequate information during the 5-year follow up from the SEER database. Among these participants, 116 died at a median age of 58, with male and female patients accounting for 67% and 33% of the deaths, respectively. The number of patients diagnosed in early years was much smaller than that of patients diagnosed in recent years. Since the data was collected in the U.S., the cohort mostly comprised of White Americans. All patients were classified by the Ann Arbor staging system used for classifying lymphomas (1983+). The number of patients in stage I and II were more than twice compared with that in the advanced stage III and IV . Detailed distributions of all cases classified by sex, race, stage, calendar year of diagnosis, and age at diagnosis are presented in P = 0.038; P < 0.001). The median survival duration was 23 months (for patients < 60-year-old) and 7 months (for patients \u2265 60-year-old) . FurtherThe survival rates for 2-year, 3-year and 10-year follow up were 39.9%, 33.7%, and 28.8%, respectively . The surP < 0.001) was associated with worse survival or COD (cause of death). Only 107 had adequate information on COD, sex, Ann Arbor Stage, age at diagnosis, and year of diagnosis. Since all death cases in the African American group could be attributable to NHL, and not to any other cause, we considered these samples to be inadequate to conduct competing risk method analysis.Tables Figs Extra nodal natural killer/T-cell lymphoma, nasal type (ENKTL-NT), is a comparatively infrequent and highly aggressive lymphoma that currently lacks optimal treatment and shows poor outcome. In our sThe current retrospective cohort study illustrated that although the most common cause of death with ENKTL-NT was NHL, there is considerable risk of other malignant cancers, including the cancers of nasopharynx, nose, nasal cavity and middle ear, lung and bronchus, myeloma and miscellaneous malignant cancer. Additionally, several factors, including age, sex, stage and time of diagnosis, could be risk factors affecting cause-specific mortality.The competing-risks regression model was applied in this study to evaluate the relationship of variables to cause-specific failures, while thDeath rate attributed to causes other than NHL was much higher for patients in advanced stage of the disease. This could be because patients in the advanced stage are more likely to face aggressive therapies, leading to potential risk of hematologic toxicity. For instance, SMILE regimen, which is the current standard to treat advanced-stage ENKTL, may be associated with more severe hematological toxicity compared with other L-asparaginase combinations and gemcitabine-based or CCRT (concurrent chemoradiotherapy) based regimens. SMILE, wPatients diagnosed in this century showed a higher survival rate than those diagnosed in the last century, partly attributed to the standardization of suitable and effective therapies, the development of HSCT, and immuThe current study has some limitations. First, some confounding factors may be included due to the retrospective nature of the study. Since we evaluated only the data collected from SEER, other potential risk factors may be missed. Second, a selection bias exists since patients with incomplete information were excluded. Third, there was a lack of consistency in death certificates obtained from different registries. In addition, since ENKTL-NT is a rare subtype of lymphoma, it is difficult to get larger sample pool to decrease the sampling error.Currently, IPI has been most widely used in NHL prognosis since more than twenty years. However,Our study indicates that the female sex, diagnosis at young age and with early stage of the disease are factors associated with better prognosis for ENKTL-NT. This population-based cohort study revealed that the most common cause of mortality in cases with ENKTL-NT is NHL, rather than other malignant cancers, diseases of heart, infections or other causes. Our study emphasizes that more effort should be made to identify patients in early stages so as to implement comprehensive treatment and prevent the occurrence of high-risk factors. Furthermore, the Ann Arbor staging and calendar year of diagnosis provide references for cause-specific death, and might help during the clinical procedures to decrease the overall and cause-specific mortality rate.www.seer.cancer.gov) which is supported by the Surveillance Research Program (SRP) in NCI's Division of Cancer Control and Population Sciences (DCCPS).Because of the sensitive nature of the data, SEER Data Use Agreement (https://seer.cancer.gov/seertrack/data/request/) is requested for everyone before accessing to the database through SEER*Stat's Client-Server Mode (https://seer.cancer.gov/seerstat/).The datasets analyzed are available in the Surveillance, Epidemiology, and End Results (SEER) program (www.seer.cancer.gov/seerstat).Since the database used was a group of public sets, it was exempted from institutional review boards. The participants were requested using the National Cancer Institute SEER*stat software version 8.3.4 ("} +{"text": "MURE\u0218AN (was delivering the welcome speech at that moment) - EFNR President, Co-Chair EAN Scientific Panel Neurorehabilitation; LEONARD SHEUNG WAI LI (Hong Kong) - President of World Federation for NeuroRehabilitation; DAVID C. GOOD (USA) - Founding Chair of Neurology at the Milton S. Hershey Medical Center of Penn State College of Medicine; VOLKER H\u00d6MBERG (GERMANY) - Program Chairman, EFNR Secretary General and WFNR Secretary General; KARIN DISERENS (Switzerland) - Acute Neurorehabilitation Unit, Neurology, Department of and Clinical Neurosciences, University Hospital of Lausanne; and ALLA GUEKHT (Russia) - Professor of Neurology, Russian National Research Medical University Director, Moscow Research and Clinical Center for Neuropsychiatry, Moscow.Members of the presidium of Leonard Sheung Wai Li (Hong Kong) presented the topic \u201cDiagnosis and management of hemiplegic shoulder pain\u201d. He works in Hong Kong as Director of Neurological Rehabilitation Centre of Virtus Medical Group, taking position also as Honorary Clinical Professor of the Department of Medicine, University of Hong Kong and Adjunct Professor of the Department of Rehabilitation Sciences of Hong Kong Polytechnic University.DAFIN F. MURE\u0218AN (Romania) presented the topic \u201cBiomarkers of rehabilitation after stroke\u201d. He is Professor of Neurology, Senior Neurologist, Chairman of the Neurosciences Department, Faculty of Medicine, \u201cIuliu Hatieganu\u201d University of Medicine and Pharmacy Cluj-Napoca, President of the European Federation of Neurorehabilitation Societies (EFNR), Co-Chair EAN Scientific Panel Neurorehabilitation, Past President of the Romanian Society of Neurology, President of the Society for the Study of Neuroprotection and Neuroplasticity (SSNN), member of the Academy of Medical Sciences, Romania, secretary of its branch in Cluj and member of 17 scientific international societies and 10 national ones, being part of the executive board of most of these societies.The following events also offered the unique opportunity of sharing the latest breakthroughs with more connected interactions and experiences, building audience involvement, while discussing innovations and concerns adopted in Neurology.\u201cThe human brain is by far the most complex physical object known to us in the entire cosmos\u201d While neurology is considered a practice of medicine dealing with disorders of the nervous system and with conditions, disease and, sometimes, rare diseases affecting the central and peripheral nervous systems , neurorehabilitation, as a complex medical process, which has the aim of recovering after nervous system injuries, promotes the necessary skills to empower the patient to work at the highest level of independence, also encouraging to rebuild self-esteem.patient-focused with customized advanced healthcare strategies to bring the patient empowerment to another level; participatory, meaning that the patient and his family are actively involved during the treatment; and community-focused, when finding the best solutions adapted for the community reintegration is considered.In this context, neurorehabilitation should also be Quality of Life (QoL) represents the well-being of individuals and societies in general, highlighting the positive and negative aspects of life, in contrast to the Health related quality of life (HRQoL), which concentrates on the effects of illness and the impact that the treatment may have on Quality of Life. Thus, QoL is broader than HRQoL because of the inclusion of non-health related aspects of life, while HRQoL is directly connected to a persons\u2019 status concerning its health or disease. In addition, the Neuro-Quality of life (Neuro-QoL), as a multidimensional patient-reported outcome measurement system, assesses conditions of mental, physical and social health that are identified, having also the ability to conduct comparisons for both PD-specific and cross-disease : 725\u2013733. Published online 2016 Feb 26. doi: 10.1002/mds.26546 (PubMed). Neuro-QoL has been developed and validated for the most common neurological conditions and created for flexible administration, such as pen and paper, interview or web-based .National Institute for Neurological Disorders and Stroke in order to create clinically-relevant health-related quality of life and psychometrically-sound measurement tools for patients who have neurological conditions or disorders like amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), muscular dystrophy (MD), epilepsy, Parkinson\u2019s disease (PD) or stroke.The instruments used in Neuro-QoL have been developed as a result of a collective research initiative sponsored by the http://www.healthmeasures.net/explore-measurement-systems/neuro-qol).From the beginning of the advancement and acceptance of Neuro-QoL measures, they have been enlarged for use in military deployment\u2013related traumatic brain injury (MDR-TBI), traumatic brain injury (TBI), Huntington\u2019s disease (HD) and spinal cord injury (SCI) :254-9. doi: 10.1089/neu.2015.3930. Epub 2015 Jul 20. (PubMed).Research concerning Neuro-QoL measures continues to spread, according to a 2016 study conducted by Research results acknowledged that \u201cnegative consequences of alcohol use\u201d was the only outcome associated positively with the sub-concussive blows. Differently, participants with a concussion history, expressed worse self-reported health .The research highlighted that athletes having a concussion history might experience bad health outcomes at a later age, but the participation in collegiate level Division III collision sports (in itself) has no risk factors concerning bad long-term neurobehavioral results."} +{"text": "Ever-growing globalization and industrialization put forward impending requirements for green and sustainable logistics (G&SL). Over the past decades, G&SL initiatives triggered worldwide deliberations, aiming at easing negative transport externalities and improving supply chain performance. This review-based paper attempts to offer a joint quantitative and qualitative understanding for the overall evolutionary trend, knowledge structure, and literature gaps of the G&SL research field. Employing the science mapping approach, a total of 306 major paper published from 1999 to 2019 were retrieved, elaborated on, and synthesized. Visualized statistics regarding publication years, journal allocation/co-citation, inter-country/institution collaboration, influential articles, co-occurred keywords, and time view clusters of research themes were analyzed bibliographically. On this basis, a total of 50 sub-branches of G&SL knowledge were classified and thematically discussed based on five alignments, namely (i) social-environmental-economic research, (ii) planning, policy and management, (iii) application and practice, (iv) technology, and (v) operations research. Finally, the current knowledge obstacles and the future research opportunities were suggested. The findings contribute to portray a systematic intellectual prospect for the state quo, hotspots, and academic frontiers of G&SL research. Moreover, it provides researchers and practitioners with heuristic thoughts to govern transportation ecology and logistics service quality. Sustainable development has inspired many green and sustainable logistics (G&SL) activities to reduce the negative effects of freight transportation and imprThe operation management of physical distribution is one of the most significant and challenging sub-issues of the macro supply chain management (SCM) , becausePrevious studies reviewed G&SL from different perspectives. By reviewing 115 papers, Zhang et al. analyzedHowever, based on the time of publication and the number of papers contained, the existing studies are outdated and incomplete, unable to provide a comprehensive analysis of the booming G&SL research in the past two years. Also, it is more difficult to integrate the multitudinous research directions to build a complete knowledge structure for G&SL. Therefore, it is of great theoretical and practical significance to objectively and quantitatively investigate the overall progress of G&SL.This study aims to conduct a comprehensive review of the global G&SL literature, so as to explore the state-of-the-art, hotspots and research trend, as well as to build the G&SL knowledge classification system. Specifically, first, tracking and analyzing the evolution of the G&SL research field from (i) publication year and journals; (ii) countries, regions, and organizations; (iii) influential documents; (iv) keywords clustering and research themes. Second, establishing the knowledge taxonomy based on the scientometric results. Third, identifying the research gaps and the future research opportunities.The novelty of this study lies in two aspects. One is to integrate the science mapping approach into the systematic literature review process to visualize the relationships among the G&SL literature. Science mapping approach is composed of data mining and bibliographic analysis, which can minimize subjective arbitrariness and grasp useful information to facilitate in-depth thematic analysis. Another is that this study further extends the bibliography to illuminate the emerging knowledge branches, gaps, and agendas in G&SL research, which will contribute to the improvement of G&SL practice and research innovation. The findings are expected to provide researchers and practitioners with a panoramic description and in-depth understanding of G&SL research. Additionally, the proposed knowledge structure can also be used as a handbook-like tool to further collect, analyze, and expand knowledge in the G&SL field and to provide references for other innovative logistics initiatives.The rest is organized as follows. In This review-based study conducted a systematic investigation on the academic development of global G&SL research with the aids of science mapping. Science mapping is a quantitative analysis approach that uses mathematical statistics and visualization techniques to study bibliographic networks in a specific field . This apIn step 1, the statistics was obtained after a comprehensive retrieval from two electronic databases, Web of Science (WoS) core and Scopus. Two rounds of selection were then performed to refine, classify, and encode the documents. The year publication trend, journal allocation and the most cited articles were described.Journal co-citation analysis: to identify the most cited journals and the research domains they belong to. This analysis helps to reveal the distribution of published journals and cited journals of the reviewed documents, so as to identify popular journals in G&SL research domain. (ii) Countries/organizations collaboration analysis: to visualize the collaborative research network of G&SL among countries and organizations, so that the readers can quickly understand the partnerships between major research communities and institutions around the world. (iii) Document co-citation analysis: to highlight the influential G&SL articles and the corresponding reference relationships. By analysis of the papers with high citation, the emerging trend of scholars\u2019 research interest to G&SL is easier to grasp. (iv) Keywords co-occurrence analysis: to map out the co-occurred time zone of the hotspots G&SL keywords and cluster them into several research themes. Network analysis of co-occurred keywords is used to clarify the knowledge structure of G&SL as well as to present the research hotspots and potential research opportunities in the future.Four scientometric tests were carried out in step 2, namely (i) In step 3, the hierarchical knowledge structure of G&SL was proposed for thematic discussion.ijc is the co-occurred or co-cited times of item i and item j, iW and jW denote the node sizes of item i and item j respectively [ix and jx are the positions of the nodes.The text mining software VOSviewer was adopted for science mapping, combining with another software CiteSpace to portray the time view of the clustered keywords based on the same data. VOSviewer, developed by van Eck and Waltman , is a coectively . The stoectively , which cFor a detailed operation manual of bibliographical experiments using science mapping approach, readers are advised to refer Jin et al. and Hu eThe advanced retrieval function in Scopus and WoS core collection database was used to retrieve the G&SL related papers published during 1999 to August 2019 see . To ensuSustainability ranks first , followed by Journal of Cleaner Production , Transportation Research Part D: Transport and Environment and International Journal of Production Economics . Among the top 15 journals, eight are from UK, four from The Netherlands, two from Switzerland, and one from Germany. The papers are mainly distributed in the three academic fields of environment, traffic engineering and operations management, but they obviously account for a larger proportion in the environmental science and sustainable field, which is in line with the connotation of G&SL.All 306 documents were found in 81 different journals. As shown in European Journal of Operational Research , International Journal of Production Research , Expert Systems with Applications , Omega and Computers & Operations Research , which can offer quantitative methods for the decision-making and optimization issues related to G&SL. The second cluster is transportation research (TR), such as Transportation Research Part A , Part D , Part E , and Journal of Transport Geography , which accumulates enormous knowledge towards transportation planning, technology and operations that can enlighten G&SL research from real-life transport demand and practice. The third cluster, including Supply Chain Management , International Journal of Physical Distribution & Logistics Management and Journal of Business Logistics , etc., reveals that a large amount of G&SL research was conducted based on the research foundation of logistics and supply chain management (SCM). Among all the publications, Journal of Cleaner Production and International Journal of Production Economics are the two most co-cited journals. They often act as hubs, integrating the results of OR, TR and SCM with social, environment or economic implications to provide cross-domain knowledge crucial to the diverse development of G&SL.As shown in According to Two evidence can be observed from Second, the international collaboration is not significant. Taking mainland China for instance, about 70 percent of 49 publications are completed entirely by domestic institutions. The Swedish publications do not have any co-authors from other countries or regions. This phenomenon may be due to the large differences in the background and model of G&SL development in different countries . MoreoveAmong the 402 organizations that contributed to G&SL research, those with more than five documents and over 30 citations were built into a network of 22 items and 22 links, as shown in Through the document co-citation test of the portfolio, the most influential G&SL publications in the past two decades were analyzed and the co-citation network was constructed. In VOSviewer, the minimum number of citations was set to 30 to build a co-cited visual network map of 83 items and 350, as shown in The top 15 most cited papers are presented in The keywords co-occurrence analysis was conducted to describe the internal composition and structure of G&SL and to reveal the frontiers . The optKeyword co-occurrence network is a static expression of a particular area that does not take into account changes over time in the manner that the terms are used . Figure Through the aforementioned analysis, the research progress, evolutionary trend, and hot-discussed topics of global G&SL are clarified. However, the generic scientometric results cannot accurately reflect the explicit division of the multifarious knowledge of a domain . Based oNearly a quarter of the literature (71 out of 306 papers) focused on evaluating and quantifying how the potential green logistics initiatives improve the \u201ctriple bottom line\u201d of existing freight activities. The subjects of these studies were basically originated from four aspects: carbon emission, energy consumption, social sustainability, and external cost-and-benefit. Mattila and Antikainen providedAnother part of emphasis was given to SEE performance of G&SL based on logistics operations and business. Guo and Ma evaluateExcept for the three-dimensional evaluation system, some scholars also analyzed the critical success factors and barriers for G&SL initiative implementation from the SEE perspectives. For instance, Arslan and Sar found thThis knowledge branch focuses on two basic G&SL topics, (i) the planning, development, and policymaking from industrial level, and (ii) the collaboration strategy and management from project level. For the former, Lindholm and Blinge indicateFor the latter, the collaboration and game among logistics service providers (LSP), government, shippers, and enterprises are paid more attention. Commonly, a positive cooperation strategy of stakeholders will significantly improve the operational performance of G&SL and evenFurthermore, several novel business and operational modes of logistics aiming at improving the sustainability in transportation process were proposed, e.g., freight consolidation , smart lOver the past decade, research on the G&SL practices were carried out over a broad range, including SCM, reverse logistics (RL), e-commerce, urban distribution, multimodal transport, and other dedicated logistics such as food and manuReverse logistics is convincingly one of the most efficient solutions to reduce environmental pollution and waste of resources by capturing and recovering the values of the used products . LegislaThe unsustainability of urban logistics makes it the most urgent goal of greening. Huge logistics demand, such as rapid business-to-business and business-to-customer logistics activities, make freight transportation in big cities face the dilemma of air pollution, poor accessibility, and livability . The praCompared with G&SL in urban domain, the sustainability issues regarding inter-city or regional logistics are more emphasized on the intermodal application. The shift of road-based modal to other transportation system, such as rail and water has the potentials of ensuring environmental sustainability, flexibility, and cost reduction . HoweverDeveloping advanced facilities and technologies is a sustainable and forward-looking solution to meet the challenge of freight transport. Many emerging logistics systems were proposed in recent years. Such as urban consolidation center , electriElectric vehicles (EVs) technology, which has been widely applied in passenger transport, is also waving a revolution in the field of G&SL. Current research on freight EVs mostly focuses on energy efficiency , fleet oFor reducing the negative externalities such as traffic congestion and disturbance, another interesting concept, i.e., transferring the ground logistics process to underground space, namely the Underground Logistics System (ULS), has aroused increasing attention. ULS refers to using a group of hierarchical underground nodes, pipelines, and tunnels to distribute cargo flows in and between cities with 24-h automated operations . ULS canThe operations research (OR) of G&SL issues that are originated from real-world applications is always being a well-concerned topic because it is directly related to the quality of some critical decision-making in logistics operation. The OR method applied for G&SL is defined as a better of science to identify the trade-offs between environmental aspects and costs, so that the corresponding decisions such as location, transportation, warehousing, and inventory can be optimized and the limited resources can be reasonably assigned . Dekker Through the above scientometric analysis and thematic discussion, the comprehensive research trend, mainstream academic topics, and knowledge taxonomy of G&SL domain were revealed. Although researchers and practitioners achieved substantial results in promoting G&SL theory and practice, there are still some shortcomings that need to be elaborated in future studies.In terms of research model, international cooperation is still lacking. The broad applicability of most G&SL knowledge based on local cases deserves further discussion, such as planning methods and evaluation systems. European countries made great efforts in rebuilding the integration of green logistics. However, the lack of international cooperation and universal solutions hinders the dissemination and deepening of knowledge, and the current achievements are far from enough to promote the globalization of G&SL, which is reflected in the imbalance of global G&SL practice.To fill this gap, although it is recognized that logistics policy has a strong regional character, cross-institutional and cross-national collaborative research on market operation, industrial metrics, technology innovation and macro development strategies should be strengthened under the trend of supply chain globalization. For example, more attention can be paid to the horizontal comparison of green logistics mode, scheme and performance under different case backgrounds. Additionally, more empirical studies are needed to be carried out in some developing countries in Asia and elsewhere in the world, considering they are the fast growing economies with higher population and logistics demand.Although the knowledge branch of research is flourishing, it is acknowledged that there is still a need to supplement the overall or holistic research to improve the knowledge system of G&SL. Research on sustainability and green has always been complex and multi-variable, interactive, with far-reaching implications. Besides, sustainability and green are public and social issues. Current theoretical applications are limited to the analysis of local or one-way relationships, such as LSP/retailer/carrier responses to green policies, planning and performance evaluation of green and sustainable initiatives.The operation and decision-making of G&SL involve many stakeholders, such as local authorities, manufactures, LSP, carriers, customers, and even the sharers of transportation resources. The impact of G&SL should also be long-term and dynamic. Thus, the whole picture includes multiple perspectives, such as the dynamic evaluation of the whole life-cycle of green logistics practice, the decision interaction among multiple stakeholders, and the follow-up research and report on a new green technology or practice.Without green innovation technologies, the effect of implementing G&SL from a management perspective alone is minimal. However, it takes a lot of time for some innovative technologies that can fundamentally improve the negative effects of logistics to move from laboratory to application. Applications such as the EV took decades to implement . AlthougThe introduction of a new thing does require a long period of demonstration, such as the reliability of the technology, the acceptability of the market and the ambiguity of the real benefits. However, the problem is often the gap and lag in the research of application management in the transition from technical problems to market application and practice management. Therefore, building effective platforms based on multidisciplinary, cross-organizational collaboration to accelerate the research and application of innovative technologies is particularly important for G&SL practices, such as ULS, RL, and CS. Such calls are all the more urgent in their own research.The concept of green and sustainable logistics has received increasing attention and consideration government sectors, scholars, practitioners, and international organizations. A large amount of practical achievement was made at both the industrial and theoretical levels. This study reviewed 306 valuable contributions regarding G&LS over the past two decades through a three-step review program. They were described in year publication, journal allocation and citation counts. Then, the bibliographic networks of countries, organizations, journal and document co-citations, keyword co-occurrence and timezone clusters of research themes were visualized to help understand the overall research status and academic progress worldwide. Grounded in the scientometric analysis, an integrated knowledge taxonomy of the G&SL field was presented, including five major alignments and 50 sub-branches.Sustainability, Journal of Cleaner Production, Transportation Research Part D: Transport Environment and International Journal of Production Economy are the top four journals, which contributed over a quarter of all G&SL papers since 1999. The maps of journal allocation and co-cited journals show that the current research is most relevant to the environmental science and transportation science. In terms of countries, China, the United States, the UK, Sweden, and India are the major territories of G&SL research. The network across co-authored organizations and countries revealed that the collaboration among different research communities is not strong. Hence an active and robust global collaboration atmosphere has not formed yet.Results indicate that the chronological publication of G&SL shows a trend of rapid increase. The quantity of literature published in 2018 is fifteen times more than that of 10 years ago. The map of co-occurred keywords showed that the most frequently discussed G&SL themes in each cluster were sustainability and management (cluster #1), freight transportation and carbon emission (cluster #2), model and reverse logistics (cluster #3), and green supply chain and green logistics (cluster #4). The timezone view of keywords showed that articles related to collaboration, transportation planning, modal shift and stakeholder were largely published during the recent years. On this basis, the knowledge taxonomy of G&SL was manually synthesized from five aspects: (i) evaluation on SEE impacts of G&SL initiatives; (ii) planning, policy, and management research; (iii) real-world application areas and practices; (iv) emerging technologies and (v) operations research and optimization methods for G&SL decision-making.Finally, the potential roadmap for filling current research gaps was recommended, which were divided into three streams: (i) more global research collaboration should be advocated to jointly develop and supplement the comprehensive evaluation framework of G&SL performance; (ii) future research efforts could focus on the interactive and dynamic relationships among sustainable development goals, green policies and the decision-making of multiple stakeholders; (iii) the application-oriented platforms and management research for some most advanced green logistics initiatives would be highly beneficial in promoting G&SL innovation.However, it should be noted that the data used in this study was confined to those research articles and review articles that were published in the peer-reviewed journals, and they were retrieved only from the two mainstream databases considering the applicability of software. Although the indexed documents could represent most of the convictive viewpoints of G&SL research, some valuable articles that were published in other forms or included in other databases might be overlooked inevitably. To sum up, this review has great room for improvement in terms of material selection. A systematic investigation incorporating valuable conference proceedings, reports, and books in the field of green logistics or green supply chain is expected to portray a more comprehensive knowledge map for future research. Additionally, the in-depth review of the hotspot themes in G&SL domain e.g., OR application and SCM, may also contribute to multidisciplinary integration and interaction."} +{"text": "Background: On May 2016, anticipating the rainy season from June to October in Mexico, we expected an increase in cases of Zika virus (ZIKV) infections. With the goal of identifying cases of GBS associated with ZIKV infection, a prospective joint study was conducted by a reference center for neurological patients and the Secretary of Health in Mexico City from July 2016 to November 2016.Methods: Serum, cerebrospinal fluid, urine, and saliva were tested by RT-PCR for ZIKV, dengue virus, and chikungunya virus in patients referred from states with reported transmissions of ZIKV infection, and with clinical symptoms of GBS according to the Brighton Collaboration criteria. Clinical, electrophysiological, and long-term disability data were collected.Results: In the year 2016 twenty-eight patients with GBS were diagnosed at our institute. In five hospitalized patients with GBS, RT-PCR was positive to ZIKV in any collected specimen. Dengue and chikungunya RT-PCR results were negative. All five patients had areflexic flaccid weakness, and cranial nerves affected in three. Electrophysiological patterns were demyelinating in two patients and axonal in three. Three patients were discharged improved in 10 days or less, and two patients required intensive care unit admission, and completely recovered during follow-up.Conclusion: Our results are similar to those reported from the state of Veracruz, Mexico, in which out of 33 samples of urine of patients with GBS two had a positive RT-PCR for ZIKV. Simultaneous processing of serum, CSF, urine, and saliva by RT-PCR may increase the success of diagnosis of GBS associated to ZIKV. In April 2016, a report of Epidemiological Surveillance for Zika virus (ZIKV) disease in Mexico reported 93 autochthonous laboratory-confirmed cases, collected between November 2015 and February 2016, and distributed amongst eight states of the country , six patients met the criteria of suspected case of ZIKV associated to GBS and five among them, met the criteria for confirmed case of ZIKV associated to GBS . On follThe systemic symptoms that preceded the onset of GBS in our patients compared with those of other studies of GBS associated with ZIKV from Latin America and Caribbean, show that the absence of symptoms before neurological manifestation was observed in a study from Colombia in two of 42 subjects, and also it was noted that the systemic symptoms of ZIKV and the neurologic symptoms overlapped in another two patients . In a stA recent study, described a ZIKV infection-associated acute transient polyneuritis , they weWith regard to RT-PCR testing, in our five patients, simultaneous samples taken at hospital admission of serum, CSF, urine, and saliva produced 10 positive results. This small case series has several limitations. It is a study from a single center, with a reference bias, since we included patients from surrounding states, able to present for evaluation at emergency room via their own resources. We include patients from states of M\u00e9xico with transmission of ZIKV, which indicated that transmitting vectors were present. The altitude of Mexico City (2250 meters above sea level) where most of our hospitalized GBS cases come from, is considered by CDC as minimal likelihood for mosquito-borne ZIKV transmission , and onlWe conclude as other Latin American and Caribbean studies that GBS associated with ZIKV infection presents in a significant number of cases with cranial neuropathy; up to two-thirds of affected patients may require admission to ICUs; the number of cases confirmed by RT-PCR in serum or CSF is low; and urine and saliva tests show variable results or are not done in some studies. The simultaneous processing of serum, urine, and saliva by RT-PCR may increase the success of diagnosis in areas without access to lumbar punctures, and when possible, other infectious agents should be sought even in areas endemic to arboviruses. Patients suffering from GBS due to ZIKV require an early evaluation in centers with qualified neurologists and an ICU stay for optimal care.This study was approved by the ethics and research committees of the Directorate of the Secretary of Health in Mexico City and the National Institute of Neurology and Neurosurgery Manuel Velasco Su\u00e1rez, M\u00e9xico City, Mexico. All participants signed informed consent prior to participation in this study.JS-H, SP, EV, CR-M, GC, KC, JD-Q, IL-M, M-EJ-C, and PK designed the study protocol and wrote the manuscript. JS-H, EV, GC, and KC cared for the patients and collected the data. JD-Q, IL-M, M-EJ-C, CR-M, and PK were responsible for the molecular diagnostic tests. All authors reviewed and approved the final version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "SLAS Discovery showcases academic screening centers and not-for-profit translational drug discovery centers. Historically, high-throughput screening was developed by the pharmaceutical industry and until the end of the 20th century was mainly carried out within its walls. After the sequencing of the human genome, academic institutions started creating screening centers to either find tool compounds or carry out RNA interference screens to study the genome. In 2003, the U.S. National Institutes of Health (NIH) Roadmap set out a plan for creating screening centers and chemical libraries, further strengthening the trend of academic screening.1 Although many academic screening centers aim to discover tool compounds for chemical biology, an appetite has always existed for more translational projects; academic screening would hopefully produce therapeutic molecules that could be used in the clinic. The desire for novel therapeutic molecules was particularly strong for neglected and rare diseases, where a lack of economic incentive meant that therapies were not available. Like many trends, after an initial enthusiasm there came a realization that developing new drugs was challenging and that academic screening centers, while facing different difficulties compared with industrial screening laboratories, also struggled to bring drugs into the clinic.This special issue of The reasons for this struggle are numerous, but I would like to underline two major reasons. First, there is a lack of funding opportunities for bringing hit compounds forward and developing lead candidates into drug candidates. Public research funds typically do not finance such work, since it is not basic research, and the projects are at too early a stage for applying for translational grants to spin out a company. It is equally hard to find pharmaceutical partners at this stage. Typically, the pharmaceutical industry invests in projects where a proof of principle has been obtained in animals, and the effects of the molecule of interest are better understood. Second, academic screening centers typically lack the infrastructure to host, curate, and process very large, high-quality chemical collections. Given the importance of the quality of compounds entering the screening process, it is perhaps not surprising that many projects did not materialize into lead molecules that could be brought forward into the clinic.To fill the gap between academic research and drug development, translational drug discovery centers were established. These centers were created to help bridge academia and industry; to have the critical mass in terms of people, instruments, compounds, and chemistry; and to form private\u2013public partnerships (PPPs) to bring innovative compounds forward into the clinic.SLAS Discovery offers a snapshot of the research being conducted in academic screening laboratories and in translational drug discovery centers. We invited many centers to contribute and 15 laboratories answered the call. As to be expected, the 15 manuscripts submitted cover a wide range of topics. The first three manuscripts, by Franke et al.,2 Warchal et al.,3 and Janosch et al.,4 describe analytical methods for phenotypic profiling of cellular responses. Using phenotypic signatures for discovering the mode of action of compounds is a very active research field, and it is not surprising to see that all three analytical articles focus on that subject. The manuscripts of Starkuviene et al.,5 Imamura et al.,6 Colussi et al.,7 Close et al.,8 Siva et al.,9 and Wiseman et al.10 all describe novel model systems, assays, and technologies that allow the screening of large collections of molecules. This illustrates that academia is a rich source for novel assays as basic research is transformed into screens, leading to unexplored therapeutic avenues. Lastly, Baillargeon et al.,11 Moraes et al.,12 Otvos et al.,13 D\u2019Agostino et al.,14 Birchall et al.,15 and Brennecke et al.16 are reviews of the screening efforts of screening laboratories and their networks, covering a wide range of applications. These reviews showcase how expertise in different fields allows academic projects to progress toward the clinic.This special issue of The 15 laboratories that contributed to this special issue all have individual expertise and operational modes, as summarized below.Helmholtz Centre for Infection Research (HZI) in Braunschweig, Germany, started its operations in 2006, although its predecessor institute was founded in 1965.2 The institute currently has 822 employees, and the three main goals of the Chemical Biology (CBIO) Department are discovering new antibacterial and antiviral drugs, characterizing their functionality, and optimizing their properties. CBIO focuses on infection research and small molecules that can function as antimicrobial or antiviral agents, interfere with pathogenicity factors, or stimulate the immune system. The discovery of new drugs includes the development of innovative, mainly phenotypic screening assays for medium-throughput screening campaigns. At the department\u2019s disposal are approximately 30,000 compounds, of which the proprietary HZI natural product collection and a proprietary academic collection (approximately 4000 compounds) are specific features. The department is actively involved in several projects of the German Centre for Infection Research (DZIF), in the Translational Unit \u201cAntibiotics\u201d and the Translational Infrastructure \u201cAntivirals.\u201d Screening is conducted either at the HZI or at an external partner site. A medium- to high-throughput screen under biological safety level S3 or S1 conditions can be performed with a robotic pipetting system. Antibacterial or antiviral screens under S2 conditions will become operative in H1/2019. Identified active compounds are profiled against the ESKAPE panel of bacterial pathogens and against mammalian cell lines to determine the selectivity index. For mode of action studies, various functional and profiling methods are established to characterize the effect of the compounds on the target pathogen or cell line. These include membrane potential and membrane permeability, high-content imaging, impedance spectroscopy,2 transcriptomics, targeted and untargeted metabolomics, and peptide arrays as the main \u201comics\u201d technologies. Specific technologies for studying the uptake of compounds in gram-negative bacteria have been established. The department also has synthetic chemistry capabilities (approximately 12 FTE) that deal with lead generation and lead optimization by synthesis. In addition to de novo-designed drug conjugates and natural product-based lead optimization, the team advances screening actives to hit series. The most promising compounds are profiled in vivo in the animal facility of the HZI, where mouse models to study the pharmacokinetic (PK) and pharmacodynamic (PD) parameters of advanced compounds in antibacterial or antiviral infection models have been set up.The The laboratory is run under an open-access model with internal and external users under a research collaboration contract.https://www.helmholtz-hzi.de/broenstrupWebsite: mark.broenstrup@helmholtz-hzi.deEmail: Edinburgh Cancer Discovery Unit (ECDU) is an academic research group located within the Cancer Research UK Edinburgh Centre, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh in Scotland, United Kingdom.3 The ECDU was founded in 2011 as a not-for-profit activity to provide a multidisciplinary group of core skills embracing advanced technology platforms and disease models, which drive innovations in oncology drug discovery and development. The ECDU research staff currently comprises Professor Neil Carragher (director), seven senior scientists, and three full-time postgraduate PhD students. Key technologies used within the unit include high-content imaging, confocal and multiphoton confocal imaging, image analysis, reverse-phase protein array, and NanoString molecular profiling of transcriptomic and posttranslation pathway networks. The ECDU mission is to develop and apply novel genetically defined 2D and 3D cell- and tissue-based assays that represent advances over the current state of the art in disease relevance and inform subsequent preclinical and clinical development strategies. The research unit is highly proficient in image-based phenotypic screening using predominantly small-molecule chemical libraries and, through partnerships, therapeutic antibodies and peptides. The laboratories are equipped with the latest kinetic and high-content screening platforms, fully integrated with plate handling robotics, barcode sample tracking, and bespoke multiparametric image analysis/informatics workflows. The unit also routinely employs both forward-phase and reverse-phase protein microarray platforms to profile preclinical and clinical samples and drug mechanism of action at transcriptomic and posttranslational pathway network levels. The ECDU works in close collaboration with several pharmaceutical and biotechnology industry partners and academic research groups to identify hit molecules, advance small-molecule lead generation, and classify compound mechanism of action through multiparametric high-content and pathway profiling. The ECDU provides an open-access model to both internal University of Edinburgh research groups and external academic or industry organizations through either fee-for-service or joint research collaboration agreements. The ECDU operates a full-cost recovery model for projects with external partners and recovery of only consumables for Cancer Research UK-funded groups within the University of Edinburgh. Intellectual property (IP) policy is flexible and dictated on a case-by-case basis dependent upon the nature of the project and research agreement . IP arrangements are pre-agreed and documented in the service or research collaboration agreements prior to commencing work with external partners. All contracts and research agreements are arranged through the business development function at Edinburgh Innovations.The https://www.ed.ac.uk/cancer-centre/impact-and-innovation/translational-science/edinburgh-cancer-discovery-unit-ecduWebsite: edinburgh.innovations@ed.ac.ukEmail: Technology Development Studio (TDS) is an academic screening facility that was created in 2004 at the Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG) in Dresden, Germany.4 The mission of the facility is to provide state-of-the-art cellular screening services for its users and to develop novel technologies where required (hence the name of the facility). The facility specializes in high-throughput microscopy since this technology provides great flexibility and allows in-depth analysis of biological systems. Homogeneous assays such as luminescence or fluorometric measurements and biochemical assays are also run when required. Both genomic and chemical screens can be run, and the facility has several genome-wide RNAi libraries and approximately 130,000 compounds. The TDS has screened a wide diversity of cellular systems ranging from simple immortalized cell lines to primary cells and stem cells both in 2D and 3D. Additionally, the facility has also screened small model organisms such as Caenorhabditis elegans and zebrafish. Protocols have been developed for handling 3D nonadherent objects with standard liquid handlers. Furthermore, optical clearing protocols and methods for imaging 3D objects in 384-well plates have been optimized. Like all facilities at the MPI-CBG, the TDS offers its excess capacity to outside users and has carried out screens for many academic and industrial collaborators not associated with the MPI-CBG. Distribution of intellectual property is decided on case-by-case by the users based on the contribution of each of the parties involved. To provide screening services to outside clients, a full-cost accounting system is used that considers salaries, consumables, instrument time, depreciation, and overheads. The TDS helps prepare grants and participates in funding calls to cover the cost of screening. Services range from designing a screening assay from scratch to executing an already optimized assay and running the analysis of the data on the institute\u2019s computer cluster. The data are owned by the client and transferred at the end of the project. To keep the costs minimal, the TDS uses open-source software for its work. Image analysis is mainly carried out with CellProfiler, Fiji, and sometimes bespoke Python image analysis scripts developed for challenging applications. For data mining, the TDS has developed a suite of software tools in the KNIME software platform. KNIME allows building analytical pipelines in a user-friendly graphical interface that helps to visualize the flow of data. The TDS has developed many tools specific for screening applications, such as plate viewers, normalization nodes, data annotations, and population analysis nodes, which can be downloaded from the KNIME website (https://www.knime.com/downloads/download-knime). One very powerful tool that the TDS introduced to the KNIME platform is scripting nodes for R and Python. These allow users to write their code in those programming languages and, with the insertion of a few lines into the code, to generate a graphical user interface in KNIME using RGG (R GUI Generator). In this manner, a computer scientist can rapidly deploy code to scientists who are not comfortable with programming scripts.The http://www.mpi-cbg.de/facilities/profiles/ht-tds.htmlWebsite: bickle@mpi-cbg.deEmail: Pharmacy Chemical Biology Center (PCBC) in the University of Pittsburgh (Pitt) School of Pharmacy was founded in 2011, evolving out of a high-throughput screening (HTS) center that was founded in 2005 with resources from the Schools of Medicine, Pharmacy, and Arts and Science.8 The Pittsburgh Molecular Library Screening Center (2005\u20132008) and the Pittsburgh Specialized Application Center (2009\u20132012) were partly supported by funding from the National Institute of Health\u2019s (NIH) Roadmap Initiative Molecular Library Pilot Screening Center Network and the National Cancer Institute\u2019s (NCI) Chemical Biology Consortium, respectively. The PCBC was created as part of the School of Pharmacy\u2019s D4 initiative to provide one-stop access to drug discovery, development, and delivery expertise in an interactive and collaborative environment. Research faculty in the D4 team have a proven track record in drug discovery and preclinical drug development in both the pharmaceutical and academic sectors. The faculty\u2019s complementary and overlapping capabilities encompass the drug discovery and development process and address the major causes of drug failure. Dr. Paul A. Johnston is the principal investigator (PI) of the PCBC and has 28 years of drug discovery experience in the pharmaceutical, biotechnology, and academic sectors. The PCBC is staffed by three full-time research scientists and varying numbers of graduate students or visiting scientists, most recently two PhD students, one MS student, and one visiting postdoc. The Molecular Devices SpectraMax M5e and Envision microtiter plate reader platforms provide multimode assay detection capabilities for UV/Vis absorbance, fluorescence intensity, fluorescence polarization, time-resolved fluorescence resonance energy transfer (TR-FRET), homogenous time-resolved fluorescence (HTRF), and luminescence. The Molecular Devices ImageXpress Micro field-based automated high-content screening (HCS) imaging platform, MetaXpress Imaging and Analysis software, and MDCStore database allow for the capture, analysis, and storage of images acquired in transmitted light and/or five fluorescent channels. The PCBC uses the ScreenAble laboratory information management system software to process and analyze compound information and HTS/HCS data. The PCBC has a 10,000-compound nonpeptide peptido-mimetic diversity subset of a 142,000 protein\u2013protein interaction library, and a 50,000-compound diversity library selected from a 410,000 core library, which enables it to effectively sample a compound diversity of 635,500 compounds. The PCBC provides guidance in assay format selection and assistance with assay development, optimization, and implementation of target-based biochemical and cell-based HTS assays, HCS imaging assays, and phenotypic drug discovery screens. The PCBC performs active confirmation, counterscreens in orthogonal assay formats, hit characterization in secondary and tertiary assays, mechanism of action studies, and bioassay support for medicinal chemistry lead optimization efforts. Projects can be loaded into the PCBC portfolio at any of these stages and are supported through a variety of funding sources, including NIH grants awarded to the PI or his collaborators at Pitt or other academic institutions , donations from philanthropic foundations, and research contracts with other academic institutions and biotechnology or pharmaceutical companies. Project budgets include personnel salaries and benefits, reagents, consumables, and Pitt institutional indirect costs. Depending upon the funding source, any unassigned intellectual property rights are subject to negotiation with the Pitt office of research.The http://www.pharmacy.pitt.edu/directory/profile.php?profile=856Website: paj18@pitt.eduEmail: Stem Cell Hotel is an innovative collaborative phenotyping unit located within the Centre for Stem Cells and Regenerative Medicine (CSCRM) at King\u2019s College London, United Kingdom.10 It is based at the 28th Floor of Guy\u2019s Tower, Great Maze Pond, with spectacular views over the Thames and the city. The center was inaugurated in December 2015 and the Stem Cell Hotel started its operation. A dedicated team of stem cell scientists, imaging experts, and analysts, with help from interns, bioinformaticians, and business advisors, is forming around the project, spearheaded by Dr. Davide Danovi. Technologies from several providers enable microscopy and high-content analysis, cell-based assays, and data integration. The Stem Cell Hotel offers access to high-content imaging and quantitative phase imaging devices. Resources and expertise in the areas of stem cell biology, artificial microenvironments for cell culture, and high-content imaging are provided as services. This includes assistance for assay development, image acquisition, and dedicated data analysis and integration. The Stem Cell Hotel develops robust methods for profiling and benchmarking cells for cell therapy and drug discovery applications. It uses dynamic and endpoint imaging and high-content analysis integrated with genomics and other biological datasets. The operation also leverages expertise from a critical mass of scientists and innovative research projects currently ongoing at the center, such as the development of standard methods for characterization of induced pluripotent stem cells (iPSCs). Created within the framework of the Human Induced Pluripotent Stem Cell Initiative (HIPSCI) and serving the UK Regenerative Medicine Platform (UKRMP), the facility offers external users services ranging from initial training on the instruments to more in-depth assistance in assay development, acquisition, and further data analysis. Its cost model varies from pure charging for the time of use of instruments and consumables to scientific collaborations, from co-development of software and applications with technology providers to contract research-type projects. Importantly, the facility works effectively with technology providers embedding instruments and technologies in the space. This has taken the form of leases, extended demos, beta testing of software, and agreements to offer the possibility to showcase devices to future potential customers. These innovative options foster a constructive dialogue and offer a testing bed for research and industry to understand needs and mature products and solutions. As an example, one of the center\u2019s technology providers can establish a strategic partnership to provide in-house technical support with the use of the entire set of instruments, ranging from assay development to image analysis. The Stem Cell Hotel\u2019s policy sees intellectual property (IP) staying with the user unless otherwise discussed on specific projects that require significant input from the Stem Cell Hotel. Born from research, boldly translational, and embracing the spirit of open innovation, the SCH grants access to state-of-the-art technology and serves communities centered around academic, clinical, and commercial research in a highly collaborative environment.The http://www.kclstemcellhotel.orgWebsite: davide.danovi@kcl.ac.ukEmail: CellNetworks Advanced Biological Screening Core Facility was established in 2007 as one of the first CellNetworks Core Facilities and is located at Heidelberg University in the BioQuant Center for \u201cQuantitative Analysis of Molecular and Cellular Biosystems\u201d in Heidelberg, Germany.5 The facility is equipped with state-of-the art instruments, and its experts possess long-term experience in the field of biologicals, RNAi screening, automated screening microscopy, and data analysis, and lately, CRISPR-mediated gene editing. The facility offers support in assay development, automated sample preparation, and high-throughput solid-phase-based transfection in multiwell plates or cell microarrays. A number of focused and genome-wide libraries of siRNAs, microRNAs, cDNAs, and crRNAs are available. In addition, the facility facilitates the contact between customers and experts on the campus to help establish collaborations and strengthen the research network in the field of biological screening in Heidelberg. In this manner, pilot projects starting out as services within the facility are developed into successful multilateral collaborations. The facility has either been a partner with or played a leading role in several projects funded by the European Union, German Federal Ministry of Education and Research (BMBF), and Baden W\u00fcrttemberg Stiftung. Recently, the High-Content Analysis of the Cell (HiCell) group was established with the aim to design, develop, and apply novel technologies for high-content screening and analysis. Once tested and standardized, the novel technologies are incorporated into the portfolio of the Advanced Biological Screening Facility and become accessible for users. HiCell focuses on the development of methodologies to interfere with cell function on DNA, mRNA, and protein levels, on the miniaturization of cellular assays such as the cell microarrays presented in this issue,5 and on combinatorial assays. The facility also offers correlative microscopy combining high-speed and super-resolution imaging as well as 3D assays and imaging. Lately, it has been focusing on single-cell analysis.The Websites:http://www.bioquant.uni-heidelberg.de/index.php?id=42Advanced Biological Screening Facility: http://www.bioquant.uni-heidelberg.de/research/groups/high-content-analysis-of-the-cell-hicell.htmlHigh-Content Analysis of the Cell (HiCell): holger.erfle@bioquant.uni-heidelberg.deEmail: Drug Discovery Initiative at the University of Tokyo, Japan, aims to promote academic research and innovation in drug discovery. It supports academic and industrial researchers who want to screen chemical samples to find either chemical biological tools, drug leads, or agrochemicals.6 The Drug Discovery Initiative has constructed a chemical library consisting of about 280,000 samples chosen primarily on druggability and structural diversity. The collection includes 63,000 samples deposited by industry since 2006. These chemicals are provided (in assay-ready plates if required) to researchers in Japan who are willing to disclose their research goals and report their assay results to the initiative under a confidentiality agreement. The users are required to pay the consumables and shipping costs, while the chemical samples themselves are free of charge. The initiative does not claim any rights to the results of the screens in the absence of intellectual contribution. The initiative has provided more than 22 million samples to more than 500 users so far. The initiative has various screening instruments that are available to users. Consultation and technical assistance can also be provided as the majority of the 25 employees have research experience in pharmaceutical companies. Short training courses on chemical screening are held several times a year to help beginners. More professional tutorials focusing on specific themes are also held in cooperation with the Conference on Biomolecular Screenology, where everybody, from beginner to expert, is welcome to attend. Recently our organization founded the Lead Exploration Unit, which has started a hit-to-lead synthesis service, as well as ADMET support, upon request.The https://www.ddi.u-tokyo.ac.jp/en/Website: ddiinfo@mol.f.u-tokyo.ac.jpEmail: Moulder Center for Drug Discovery Research is located at the Temple University School of Pharmacy in Philadelphia, Pennsylvania.7 The center was established in 2008 thanks to a generous gift from School of Pharmacy alumni Lonnie and Sharon Moulder. The mission of the Moulder Center is focused on the discovery of new clinical candidates. Furthermore, the center provides education and training for students, postdoctoral fellows, and faculty in the application of modern drug discovery techniques. The center participates in collaborative research within Temple University as well as externally with research institutions, universities, and the pharmaceutical industry. The Moulder Center embraces three business models: collaborative research, contract research, and consulting/providing expertise and oversight to external resources. The Moulder Center is a fully integrated academic drug discovery laboratory with resources and capabilities in assay development and high-throughput screening, in vitro pharmacology medicinal chemistry, in vitro ADME, and in vivo pharmacokinetics. The center is staffed with faculty, research associates, postdoctoral fellows, and graduate students. Most Moulder Center members are highly skilled in drug discovery and have accrued many years of experience working in the pharmaceutical industry. High-throughput screening is focused on small-molecule collections with automation supported by Janus workstations . Compound and data management utilizes the Dotmatics informatics suite . The center maintains an ADME screening panel of in vitro assays to assess drug-like properties of candidate compounds. These assays measure hERG inhibition, aqueous solubility, chemical stability, stability in physiological fluids, liver microsomal stability, hepatocyte stability, plasma protein binding, microsomal partitioning, CYP450 inhibition, and permeability. The center\u2019s consumable and personnel costs are covered by funding sources from grants and contracts. The intellectual property policy is university driven and supported by Temple University\u2019s Office of Technology Transfer.The https://moulder.temple.edu/Website: aboumag@temple.eduEmail: Chemical Biology & Therapeutics group (CBT) is located at the medical faculty of Lund University, Sweden, and the Chemical Biology Consortium (CBCS) is located at Karolinska Institutet, in Stockholm, Sweden.9 The CBT was established in 2013 with financial support from Lund University to support its researchers. The facility\u2019s personnel are funded by MultiPark and projects are managed as academic collaborations . The facility currently has Bravo BenchCel liquid handling systems with a PlateLoc micropate sealer from Agilent and MultiDrop Combi from ThermoFisher . The facility uses various technologies for screen readouts in collaborations with other infrastructures at the faculty. The large number of assay modalities allows the CBT to be very flexible in the types of assays it can process. The CBCS is a national infrastructure for chemical biology within the Science for Life Laboratory. Its operations began in 2010, first funded by the Swedish Research Council and currently by the Science for Life Laboratory and the host universities (Karolinska Institutet and Ume\u00e5 University). At CBCS, 12 scientists with a background in the pharmaceutical industry are employed. CBCS offers instrumentation and experience in performing most types of high-throughput screens . Through CBCS, academic researchers have access to ~200,000 small molecules, including a set of known drugs and annotated compounds. CBCS performs chemical screens and has suitable liquid handling equipment and technologies for most assay readouts. Access to CBCS is open on equal terms to all Swedish researchers . Swedish researchers are prioritized over international users. The cost model has the Swedish academic researchers pay for consumables and compound access. The researchers only partially pay for expertise during assay development, screening, and follow-up work. The CBCS general user agreement does specify any IP rights; however, CBCS does not intend to claim rights and has, upon request of users, transferred IP rights.The Websites:http://portal.research.lu.se/portal/en/organisations-researchgroups/chemical-biology-and-therapeutics(a49506d7-e018-4ccb-b701-f3f1ff02fcfb).htmlCBT: Roger.Olsson@med.lu.seEmail: http://www.cbcs.seCBCS: Anna-Lena.Gustavsson@ki.seEmail: Centre for Integrative Biology High-Throughput Screening and Validation (CIBIO-HTS) facility has been operating since 2011 within the University of Trento, Italy.14 CIBIO infrastructure includes nine state-of-the-art core facilities, all independent from the university\u2019s research groups and run by qualified personnel. The CIBIO-HTS facility supports the development of screening programs and offers an interactive and open environment to researchers willing to apply chemical and functional biology or image-based cytometry to their research. The facility is staffed with four people with many years of experience. Close collaboration with 40 internal research groups generated core competencies in oncology and neurobiology. Collaboration with other CIBIO core facilities allows the facility to expand its expertise to stem cells, zebrafish, and Drosophila, as well as to protein production and biophysical characterization of molecules and interactions. The facility can handle a wide variety of experimental setups, from low-throughput drug testing to higher-throughput screenings, using automated liquid handlers , BioTek EL406 , multimode plate readers , imagers , and other multi-well-based detection systems . Long and broad experience in high-content screening allows flexibility in the choice of the model to be screened, from 2D and 3D cultures to whole organisms, for both target-based investigations and multiparameter phenotypic profiling. The commercial libraries available include drugs or drug-like molecules selected for their known pharmacological and biological effect and for their chemical diversity. The CIBIO-HTS facility has acquired skills in a broad range of screening projects, both biochemical and cell based, with the focus on RNA biology and posttranscriptional processing. The reporter-based cellular assays (luciferase or fluorescent proteins) are applied to challenge the regulatory processes of mRNA, such as splicing, export, localization, turnover, and translation efficiency. Novel biochemical assays employing AlphaScreen and DMR Epic Label-Free technologies have been developed to study RNA\u2013protein interactions. Finally, the facility teams up with other internal infrastructures to provide validation strategies and techniques such as transcriptomics and proteomics and supports quantitative and droplet digital PCR and polysomal fractionation analysis. The use of an electronic booking system to track the usage of assets and the application of access rules, as well as a detailed analysis of the costs associated with the facilities, allows opening access to internal and external academic users, as well as private companies. CIBIO-HTS also provides assistance for grant applications by drafting experimental designs and providing letters of support. In case of publications or new patent applications, co-authorship or joint inventorship may be considered appropriate only if the staff have provided a significant intellectual contribution to the assay development or the hit validation.The https://www.cibio.unitn.it/48/high-throughput-screening-hts-and-validationWebsite: hts.cibio@unitn.itEmail: LifeArc, formerly known as MRC Technology, is an independent medical research charity.15 Founded in 2000, MRC Technology brought together the different groups involved in the translation of UK Medical Research Council-funded science. In 2001, the assay development group was formed to work with academic scientists to generate reagents and assays to allow plate-based screening of novel targets. It was quickly realized that to interrogate targets further, quality chemical probes were required, and that screening alone was unlikely to deliver these. In 2005, a chemistry group was added with a focus on medicinal chemistry, developing chemical tools needed for target validation and developing early-stage chemical hits into chemical leads as a basis for collaborations or licensing deals with pharma and biotech partners. This gave rise to LifeArc\u2019s Centre for Therapeutics Discovery (CTD). LifeArc now works independently of the MRC and is funded by successful outputs of licensing . Since 2016, CTD has been based in Stevenage (UK) and collaborates with academic groups around the world. There are more than 90 scientists working in early drug discovery, including target validation, pharmacology, assay development, screening, antibody engineering, medicinal chemistry, computational chemistry, and early ADME. Screening formats include cellular (target-specific and phenotypic), biochemical, and increasingly, biophysical screening. LifeArc has an in-house compound collection including a diversity and an indexing set, as well as several focused sets , fragments, natural products, annotated). LifeArc\u2019s model is to form active collaborations with academic groups in the early drug discovery space. Early interactions are key to shaping a project in terms of validation experiments, advice on assay development, and the sharing of compound libraries. When a project moves into the LifeArc portfolio, a collaboration agreement with the host institution is put in place determining governance and intellectual property ownership and a project team is formed consisting of LifeArc and originating lab personnel. LifeArc covers all the costs associated with LifeArc activities and can work with an academic group to secure funding for aspects ongoing in the originating lab.https://www.lifearc.org/working-with-us/drug-discovery/Website: info@lifearc.orgEmail: Fraunhofer-Institute IME in Hamburg, Germany, began its operations in 2007 and currently employs 25 employees.12 The IME operates two fully automated screening systems. The first system is a PerkinElmer Cell::Explorer system , which has a modular design and contains integrated liquid handling, compound reformatting, incubators, a high-throughput microscope, and a high-throughput multimode plate reader for the commonly used screening assays. The second system is the Tecan Fluent 760 , which is a compact and fast system for automated screening. The screening technologies and readouts available include AlphaScreen, time-resolved fluorescence resonance energy transfer (TR-FRET), fluorescence intensity, confocal high-content imaging, absorbance, luminescence, reporter gene, cellular biosensors , immunocytochemistry, cell migration and translocation, ion sensing , enzyme-linked immunosorbent assay (ELISA), proximity ligation analysis, cell-free electrophysiology, label-free dynamic mass redistribution, surface plasmon resonance, and fluorescence polarization. Most assays are screened against compound libraries, and we have access to some 500,000 small molecules and 10,000 natural products. The target classes the IME has experience with include kinases, G-protein-coupled receptors (GPCRs), ion channels, histone demethylases (HDACs), phosphodiesterases (PDEs), synthases, transferases, deubiquitinylases, protein\u2013protein interactions, transporters, efflux pumps, proteases, noncoding RNAs, transcription factors, nuclear hormone receptors, mitochondrial membrane channels, DNA binding proteins, and toxicity readouts . The IME operates under an open-access model with internal and external users under a research collaboration contract using a full-cost recovery model. Where appropriate, intellectual property is shared between users and the platform depending on the respective contributions and contract conditions.The https://www.ime.fraunhofer.de/en/Research_Divisions/business_fields_TM/screeningport.htmlWebsite: philip.gribbon@ime.fraunhofer.deEmail: Bio-Analytical Chemistry division at the Vrije Universiteit Amsterdam in the Netherlands, Dr. Jeroen Kool works as an analytical chemist with research interests in high-resolution screening of biologically active mixtures.13 His research achievements allow full compatibility of analytical separations with biological assays (including cellular) and parallel mass spectrometry (MS) detection for investigation of bioactive mixtures using miniaturized setups and nanospotting technologies. These techniques combining chromatography, MS, and bioassays in one analytical platform are now known as nanofractionation analytics. In recent years, hyphenated analytics for both liquid chromatography (LC) and gas chromatography (GC) separations to bioassays for the identification of biologically active toxicants in natural extracts, food, and the environment have been developed. For GC fractionations, Kool developed and patented an automated system for high-resolution fractionation of complete gas chromatograms with parallel chemical detection, such as flame ionization detection (FID) and MS. Kool also developed analytical methodologies for bioactivity profiling of metabolic mixtures from drugs and lead compounds targeting G-protein-coupled receptors (GPCRs), nuclear receptors, protein kinases, and ion channels. Currently, Kool is working on postcolumn microfluidics and nanospotting analytics for the analysis of bioactive mixtures only available in low amounts. Insect and animal venoms are examples that contain many different, highly potent, and sometimes very selective peptide ligands for a large variety of medicinal targets. Some of the analytics and other techniques in the Bio-Analytical Chemistry division include nano-LC-MS systems, nanofractionation analytics , high-resolution screening analytics, GC fractionation equipment , and LC fractionation bioassay analytics . Typically, bioassays use existing formats and implement them in the laboratory\u2019s analytics, or they are developed from scratch if they are not available. The bioassays range from enzymatic to ion channels, nuclear receptors, GPCRs, cellular assays, and other assay formats. Kool\u2019s laboratory has ample expertise in capillary electrophoresis (CE)-MS analytics and started working with matrix-assisted laser desorption/ionization (MALDI)-MS imaging projects recently. One of the research efforts is directed at trapped ion mobility spectrometry (TIMS) looking at metabolites of drugs. Fifteen MS systems (high and low resolution), 10 CEs, 3 nano-LCs, 3 ultra-pressure liquid chromatography (UPLC) systems, 40+ high-pressure liquid chromatography (HPLC) systems, several GCs, an HTS lab, a cell culturing lab, and access to a radio nucleotide lab are available. Dr. Kool\u2019s core expertise is in coupling separation techniques with MS and bioassays to identify the bioactive components in complex mixtures. The laboratory is open to discuss possibilities for any form of access/cost model, ranging from full collaborations to full fee-for-service models.Within the https://research.vu.nl/en/persons/j-koolWebsite: j.kool@vu.nlEmail: Scripps Research Molecular Screening Center (SRMSC) found its permanent home in a brand-new facility built in 2008.11 It currently comprises more than $22 million of specialized automation for early drug discovery and screening. The Translation Research Institute (TRI), under the direction of Patrick Griffin, was founded at approximately the same time. Together with researchers in the Rosen Lab in La Jolla, California, at Scripps Research, about 15 employees represent one of the most competitive academic industrial screening centers worldwide. The SRMSC houses two fully automated Kalypsys/GNF platforms, one a screening platform and the other a cherry-picking platform. Additionally, matching technologies are appended within the SRMSC to allow scientists and engineers to develop assays and prepare them for automated screening. The SRMSC is a small-molecule screening center and archives more than 1 million compounds, including a proprietary collection of >665,000 molecules derived from commercial sources from around the world. As part of this collection, medicinal chemistry has included ~40,000 unique compounds that are not found elsewhere. These collections are screened against a myriad of disease targets, including cell-based and biochemical assays. Scripps has proven competence in a broad range of detection formats, including high-content analysis, fluorescence, bioluminescence resonance energy transfer (BRET), time-resolved fluorescence resonance energy transfer (TR-FRET), fluorescence polarization (FP), luminescence, absorbance, AlphaScreen, and Ca++ signaling, to name a few. These technologies are applied to both NIH-derived academic collaborations and small- and large-scale biotech and pharma initiatives. The SRMSC and TSRI are proud to have been able to provide full-cost recovery since their inception and are recognized for generating intellectual property impacting the benefit of mankind (see Ozanimod).After initially locating at the Florida Atlantic University Honors College in Jupiter, Florida, in 2005, the https://hts.florida.scripps.edu/Website: SRIMSC@scripps.eduEmail: EU-OPENSCREEN, the European research infrastructure for chemical biology, was founded in April 2018 with the support of 21 screening and medicinal chemistry partner sites from eight European member and observer states and the European Commission.16 EU-OPENSCREEN is headquartered in Berlin, and its main goal is to facilitate the development of new molecular research tools and drug candidates\u2014thus ensuring European competitiveness in the fields of chemical biology and early drug discovery. The EU-OPENSCREEN compound collection of 140,000 molecules can be accessed by collaborators from academia and industry and represents one key physical asset of the network. Its commercial part comprising 100,000 small molecules and fragments from both large and smaller, more specialized vendors is carefully designed and assembled by five EU-OPENSCREEN partner institutions to ensure high diversity and distinctiveness of the library . In addition, the academic part comprising up to 40,000 molecules, crowd-sourced from European chemistry laboratories over time, will further add unique chemistry and new molecule classes such as natural product-like compounds to the collection. Hereto, expertise regarding isolation and chemical characterization of marine and microbial natural products is well represented within the network. All compounds of the EU-OPENSCREEN compound collection are bioprofiled for physicochemical properties such as solubility, potential for interference with certain assay readouts, and cellular toxicity, at designated partner institutions. Bioprofiled data and structural information are stored in the open-access European Chemical Biology Database (ECBD). Incoming projects are assigned to the most suitable of the 15 EU-OPENSCREEN screening partners based on individual project needs. Due to its distributed character, EU-OPENSCREEN as a collective covers a plethora of specialized project needs alongside standard phenotypic and target-based high-throughput and high-content screening approaches. These include fragment-based screening by nuclear magnetic resonance (NMR) and surface plasmon resonance (SPR), high-throughput mass spectroscopy screening, high-throughput flow cytometry methodologies for cell-based screens, screening under BSL2/3 conditions, protein\u2013protein interaction screens, multiplexed biochemical and imaging assays, combinatorial screening of compounds, target-based biophysical screening, in vitro and in vivo ADME profiling, and virtual screening for hit analogues. Importantly, innovative cellular systems such as human organoids, induced pluripotent stem cells, or patient-derived cells can be readily supported. After the initial screen, collaborators can rely on EU-OPENSCREEN\u2019s six medicinal chemistry partners for chemical optimization of hit compounds. Expertise within the network includes the design, synthesis, and chemical optimization of compounds and fragments with a high degree of three-dimensionality and stereochemical diversity, as well as structure\u2013activity relationship analysis during chemical optimization. In conclusion, due to EU-OPENSCREEN\u2019s interdisciplinary, collaborative working model, collaborators can run their entire project within the network and benefit from the collective expertise of the 21 partner sites. Biological assay data will be made publicly available in the ECBD after a grace period of up to 3 years, which is meant to ensure publication and/or patenting of experimental results. EU-OPENSCREEN is a not-for-profit organization, and costs are calculated to only cover the replenishment of used compounds and consumables at partner institutions. Collaborators from industry will be charged a 20% surcharge. A more detailed description of the EU-OPENSCREEN network and its participating partner institutes can be found online.https://www.eu-openscreen.eu/Website: office@eu-openscreen.euEmail: https://www.linkedin.com/groups/4083717 and/or the SLAS European Academic Drug Discovery Special Interest Group online at https://www.linkedin.com/groups/8417733. Members of these special interest groups also host meetings during the annual SLAS International Conference and the annual SLAS Europe Conference to discuss topics of interest and network with other professionals in this field.While this special issue offers just a glimpse into the work going on in the academic and not-for-profit drug screening world, it illustrates a rich landscape that offers a great diversity and a range of expertise. For more information and to connect more closely with this exciting community, readers of this special issue are encouraged to join the SLAS Americas Academic Drug Discovery Special Interest Group online at"} +{"text": "Urban metabolism analyzes the supply and consumption of nutrition, material, energy, and other resources within cities. Food, water, and energy are critical resources for the human society and have complicated cooperative/competitive influences on each other. The management of interactive resources is essential for supply chain analysis. This research analyzes the food-water-energy system of urban metabolism for sustainable resources management. A system dynamics model is established to investigate the urban metabolism of food, water, and energy resources. This study conducts a case study of Shihmen Reservoir system, hydropower generation, paddy rice irrigation of Taoyuan and Shihmen Irrigation Associations, and water consumption in Taoyuan, New Taipei, and Hsinchu cities. The interactive influence of the food-water-energy nexus is quantified in this study; the uncertainty analysis of food, water, and energy nexus is presented. This research aims to analyze food, water, and energy resources of urban metabolism for sustainable resources management. The interactions between the natural environment and humans in cities are complex systems. Urban metabolism is a concept simulating the production, distribution, intake, digestion, and absorption of nutrition, material, energy, and other resources within cities ,5,6,7,8.2 in Beijing and Shanghai [Previous studies have investigated urban metabolism using life cycle, material flow, meta-analysis, input-output analysis, and simulation tools. The nexus of water, energy, material, carbon dioxide emission, ecological system, climate, and human society has been explored and discussed. Chen and Chen 2017) analyzed the coupling of carbon and energy flows in the cities of Beijing and Issaquah. The research used meta-analysis and nexus modeling to evaluate the energy-carbon nexus . Chen et17 analyzShanghai . Zhang eShanghai .Related studies of the urban metabolism involve large-scale, conceptual, life cycle, or simulation modeling analysis. This research aims to investigate the cooperative/competitive interaction of the water-energy-food nexus. Furthermore, quantitative analysis of the interactive influence of the food-water-energy nexus is presented in this study. A case study in a system with water supply, water consumption, hydropower generation, and paddy rice growing is also conducted. In this research, system dynamics models are established to analyze the urban metabolic food-water-energy systems. System dynamics was developed in 1950; the simulation method is widely applied in industrial, engineering, natural science, and public policy fields ,34,35,36The system dynamics models are used to examine cooperative/competitive behaviors, including the water supply, water consumption, hydropower generation, and food crops growing in food-water-energy systems. The urban metabolism simulation model is established on NetLogo platform. NetLogo system is a popular and powerful system dynamics simulation and agent-based interactive simulation platform. NetLogo models utilize system dynamics, patch, turtle, plot, and monitor agents to analyze the system performance and agent interactions. In our analysis, NetLogo uses patches and turtle agents to simulate water, paddy rice, hydropower production, and demand/supply interactions. Furthermore, monitor agents collect and display dynamics changes of food, water, and energy nexus systems.Compared with related studies, the significant contribution of this research includes the establishment of a framework for analyzing food-water-energy nexus, quantifying competition/cooperation among resources, and integrated resources management. In addition, this research constructs system dynamics models for participants in food-water-energy systems. Accordingly, the system dynamics model is established on the NetLogo platform and then equilibrium solutions of competition/cooperation are determined. Moreover, a case study of competition and cooperation among resources is conducted and discussed. This paper is organized as follows. This section begins with formulating a system dynamic model to investigate the food, water, and energy nexus. The model is capable of analyzing complex interactive component behaviors and nonlinear system performance over time. The system dynamics model of the food-water-energy system is established as follows. The water resources network model is formulated including dams, water flow, water consumption, and hydropower production. In the model, water inflow, loss, and spillage at node The model is a system dynamics simulation model with multiple objectives. Three objective functions are formulated, including food production maximization and energy production maximization. Water resources supply and demand are considered in the constraints. The objective functions are subject to constraints of water mass balance, food yield, hydropower generation, bioenergy production, reservoir operation, and non-negativity equations.Assuming The food-water-energy system dynamics model is a time-dependent simulation problem. The study applies the system dynamics simulation method to analyze the model in Equations (1)\u2013(11). The method simulates control and state variables of the system under different uncertain scenarios.This research analyzes the system dynamics model of the urban metabolic food-water-energy system in Equations (1)\u2013(11). A system dynamics model is established by using NetLogo to simulate the components interaction and system performance. Different scenarios are simulated and compared by the NetLogo system dynamics model. NetLogo is a simulation modeling platform and is able to conduct system dynamics and agent-based simulations. NetLogo is an open source software widely applied for simulations in physics, chemistry, biology, economics, social science and other related research fields. The model simulates agent-based interaction by using stationary Patch agents and mobile Turtle agents. Additionally, Link agents are used for connecting multiple agents, and Observer agents are set to record individual behavior and system performance.The research conducts a case study of the urban metabolic food-water-energy system in the Shihmen Reservoir system. The food-water-energy nexus system includes the Shihmen Reservoir water resources system, hydropower generation system, paddy rice land, and household water consumption in Taoyuan, New Taipei, and Hsinchu cities see . AccordiThis research constructs a system dynamics model. The model includes system dynamics simulation and agent-based models. 3 and a catchment area of 760 km2. The reservoir is confronted with a serious sediment accumulation problem. One-third of the active storage capacity has been taken over by sediments.In the simulation, farm land for paddy rice production is about 36,000 hectares in Taoyuan; farm land includes approximately 24,000 hectares of the Taoyuan Irrigation Association and 12,000 hectares of the Shihmen Irrigation Association. Rice production rate is about 5 ton/hectare. For the water supply system, the Shihmen Reservoir is a multi-purpose reservoir located in the upstream of Dahan River in northern Taiwan . It is t3/s and total installed capacity of 90 MW. The average annual hydropower generation is about 230,000 MWh. The maximal rice production is estimated at 180,000 tons per season for 36,000 hectares of paddy rice land of the Taoyuan Irrigation Association and the Shihmen Irrigation Association. The average water consumption of paddy rice irrigation is approximately 350 million m3.In the model, the Shihmen Reservoir supplies water to a population of 3.5 million people in Taoyuan, New Taipei, and Hsinchu cities. The Shihmen Reservoir delivers water to irrigation land for agricultural water consumption of the Taoyuan Irrigation Association and the Shihmen Irrigation Association. Agricultural water demand, on average, accounts for 45% of the total water supply of the Shihmen Reservoir while the rest goes to the domestic water demand. Furthermore, the Shihmen Reservoir has two hydropower generators, and each of them has a capacity of 45 MW. The hydropower system has a maximal water flow of 140 mIn addition, uncertainty analysis of the water inflow of reservoir, water demand, and paddy rice production are simulated using a Monte Carlo simulation in the model. Food, water, and energy are essential and scarce resources. The interactive relationship and allocation of food, water, and energy resources are crucial topics. This research analyzes the integrated food-water-energy systems for urban metabolism. The contribution of this study is to establish a framework for quantifying the cooperation/competition of multiple resources and formulating the system dynamics models of the urban metabolic food-water-energy system. The system dynamics model is capable of simulating different uncertain scenarios for uncertain analysis. A case study of food-water-energy system is conducted consisting of the Shihmen Reservoir water resources system, hydropower generation, paddy rice production of the Taoyuan and Shihmen Irrigation Associations, and domestic water supply in Taoyuan, New Taipei, and Hsinchu cities. The results quantify the interactive influence between food, water, and energy resources. In addition, stochastic simulation of water inflow shows uncertain water supply, paddy rice growing, and hydropower production systems. Future studies include simulation of uncertain climate scenarios, human behavior, hydrological process, energy availability, and energy load of nexus systems. Interactive agent-based modeling with multi-objective in food-water-energy systems are also of interest."} +{"text": "Nature Communications; 10.1038/s41467-018-06944-1; published online 29 October 2018Correction to: In the original version of this Article, Haoping Liu, who conceptualized, designed and supervised the project and acquired funding, was inadvertently omitted from the author list. Furthermore, the affiliation of Jiaxin Gao and Haoping Liu with \u2018Department of Biological Chemistry, University of California, Irvine, CA 92697, USA\u2019 was omitted. Finally, funding from NIH grant GM117111, and contributions from Dr. Li-lin Du of NIBS for providing pPB[ura4] and pDUAL-PBase and Allan Bradley of Sanger for hyPBase, were not acknowledged. These errors have now been corrected in both the PDF and HTML versions of the Article."} +{"text": "Health Policy and Planning, doi: 10.1093/heapol/czy053.The authors wish to apologize for the following error: the name of one of the co-authors, Beth Ann Hart, was incorrect as initially provided. This has now been corrected in print and online."} +{"text": "This paper presents an innovative conceptual approach to health care policy for older adults: the Age-Friendly Health Systems Integrated Interprofessional Model. In 2017, the John A. Hartford Foundation and Institute for Healthcare Improvement, in partnership with the American Hospital Association and Catholic Health Association of the United States, advanced the concept of an Age-Friendly Health System. This initiative is designed to respond to the needs of a burgeoning U.S. older adult population, expected to double from 2012 to 2050, largely due to the aging of Baby Boomers and increased life expectancy. These Baby Boomers will demand a well-coordinated, communicative health system responsive to their values and preferences. In an Age-Friendly Health System, all older adults receive the best possible care, without care-related harms, and with satisfaction of care received. Essential elements include what matters, mentation, mobility, and medications, with a focus on patient-directed, family-engaged care. While a solid framework for improving healthcare for older adults, this model is further strengthened by incorporating the essential elements of person-, family-, and community-centered approaches to care; interprofessional team-based competencies, and Quadruple Aim outcomes. This enhanced model, referred to as the Age-Friendly Health System Integrated Interprofessional Model, combines elements essential to quality healthcare within the framework of an Age-Friendly Health System. This paper will present the original Age-Friendly Health System framework, the proposed Age-Friendly Health System Integrated Interprofessional Model, then compare and contrast each model\u2019s essential principles. Implications for adoption of this enhanced model for policy, education, and practice will be explored."} +{"text": "Rapeseed is an important oilseed with proper fatty acid composition and abundant bioactive components. Canada and China are the two major rapeseed-producing countries all over the world. Meanwhile, Canada and Mongolia are major importers of rapeseed due to the great demand for rapeseed in China. To investigate the metabolites in rapeseeds from three countries, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS)-based metabolomics was employed to analyze rapeseeds from China, Canada, and Mongolia. As results, 67, 53, and 68 metabolites showed significant differences between Chinese and Canadian, Chinese and Mongolian, and Canadian and Mongolian rapeseeds, respectively. Differential metabolites were mainly distributed in the metabolic pathways including phenylpropanoid biosynthesis, flavone and flavonol biosynthesis, and ubiquinone and other terpenoid-quinone biosynthesis. Among the differential metabolites, contents of sinapate and sinapine were higher in Chinese rapeseeds, while the contents of brassicasterol, stigmasterol, and campestanol were higher in Canadian rapeseeds. These findings might provide insight into the metabolic characteristics of rapeseeds from three countries to guide processing and consumption of the products of rapeseed. Brassica napus; Cruciferae) is an important oil crop in agriculture worldwide, and ranks the third largest source of vegetable oil all over the world obtained from OPLS-DA loadings S-plot is greater than 0.8; (b) p-value calculated using t-test is less than 0.001; (c) fold change (FC) is more than twice or less than half.Data processing of data normalization was conducted as described elsewhere, with a little modification . Pareto Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation and metabolic pathway analysis of differential metabolites were performed on MetaboAnalyst 4.0 platform . KEGG meMetabolomics is commonly used to study the total metabolites of a given plant tissue ,25. In tPCA is an unsupervised method, which plays an important role in the grouping and discrimination of chemicals in food and medicines . PCA wasOPLS-DA is a rotated and powerful supervised modeling method used for the identification of variables that drive group separation. The S-plot obtained from OPLS-DA could visualize the variable influence in the model and identify statistically significant and potentially biochemically significant metabolites . To iden2 and Q2 values of the three models were close to 1.0 , suggesting that the models were of good fitness and predictability. The S-plots showed the variable importance in OPLS-DA models and could be used to identify the differential metabolites between every two countries.OPLS-DA score plots for Chinese and Canadian rapeseeds, Chinese and Mongolian rapeseeds, Canadian and Mongolian rapeseeds were used to identify important metabolites, respectively. The rapeseed samples were separated into two clusters of all groups. The results suggest that there were differential metabolites between each pair of countries. The Rt-tests together. S-plot and univariate analysis were employed to identify the differential metabolites of the three groups to ensure the accuracy and reliability of the results. The differential metabolites were identified using the criteria that the absolute value of reliability correlation [p(corr)] of S-plots was greater than 0.8, the p-value calculated using t-test was less than 0.001, and the FC was more than twice or less than half.Univariate analysis methods are the most common methods used for exploratory data analysis. Univariate analysis performed on Metaboanalyst 4.0 can provide a volcano plot that combines FC analysis and t-test threshold (y) 0.001 are displayed by volcano plots in p-values were log transformed, and the different metabolites are shown as red circles above the threshold.Important metabolites with FC threshold (x) 2 and Based on the results of the S-plots and volcano plots, the comparison between Chinese and Canadian rapeseeds was performed, and 67 metabolites out of 152 were identified as differential metabolites based on the criteria. The 67 metabolites, as listed in The pathway analysis module of MetaboAnalyst 4.0 uses high-quality KEGG metabolic pathways as the back-end knowledge, and combines the results from powerful pathway enrichment analysis with pathway topology analysis to help researchers identify the most relevant pathways involved in the conditions under study. The pathways of the differential metabolites of Chinese vs. Mongolian rapeseeds, Chinese vs. Canadian rapeseeds, and Mongolian vs. Canadian rapeseeds shown in p < 0.01).The differential metabolites between Chinese and Canadian rapeseeds were involved in 16 pathways. Based on the characteristics of the bubble plot in Canada and China are the biggest producers of rapeseeds, and the rapeseeds of the two countries are the biggest sources of rapeseeds consumed in China. The qualities of Canadian and Chinese rapeseeds attract more and more attention of processing enterprises and consumers.The metabolic pathways of flavone and flavonol biosynthesis and phenylpropanoid biosynthesis were differentially altered between Chinese and Canadian rapeseeds from the results of metabolic pathway analysis. The metabolites in the two pathways showing significant differences in the two countries include kaempferol, luteolin, coniferyl alcohol, ferulate, coniferin, sinapate, 1-O-sinapoyl-beta-D-glucose, and sinapine. The relative contents of these compounds of rapeseed collected from the two countries are shown in The relative contents of kaempferol, luteolin, coniferin, sinapate, 1-O-sinapoyl-beta-D-glucose, and sinapine of Chinese rapeseeds were significantly higher than those of Canadian rapeseeds . WhereasSinapate, commonly known as sinapic acid, is an important bioactive substance with antioxidant, anti-inflammatory, anticancer, antimutagenic, antiglycemic, neuroprotective, and antibacterial activities . SinapatApart from sinapate, stigmasterol, brassicasterol, and campestanol detected in this study are also well known for their higher contents in rapeseeds. Phytosterols share a similar structure with cholesterol and are structural components of plant membranes. Therefore, they can regulate the physicochemical properties of cell membranes to respond to abiotic and biotic stress . By inhiIn this study, UPLC-Q/TOF-MS-based comparative metabolomics combining chemometric methods was employed to study the chemical composition of Chinese, Canadian, and Mongolian rapeseeds. OPLS-DA loading S-plot and univariate analysis were employed to identify the differential metabolites of the rapeseeds collected from three countries. As results, 67, 53, and 68 differential metabolites were identified in Chinese and Canadian rapeseeds, Chinese and Mongolian rapeseeds, and Canadian and Mongolian rapeseeds, respectively. The metabolic pathway analysis results showed that phenylpropanoid biosynthesis, flavone and flavonol biosynthesis, and ubiquinone and other terpenoid-quinone biosynthesis were differentially altered. Chinese rapeseeds have higher contents of sinapate and sinapine, while Canadian rapeseeds possess higher contents of phytosterols. It is necessary for oil-processing enterprises to choose proper rapeseed materials and processing technologies to produce rapeseed oils with higher canolol or phytosterols contents."} +{"text": "Internet addiction (IA) has become a major public health problem among college students. The aim of this study was to examine the relationship between self-identity confusion and IA and the mediating effects of psychological inflexibility and experiential avoidance (PI/EA) indicators in college students. A total of 500 college students (262 women and 238 men) were recruited. Their levels of self-identity were evaluated using the Self-Concept and Identity Measure. Their levels of PI/EA were examined using the Acceptance and Action Questionnaire-II. The severity of IA was assessed using the Chen Internet Addiction Scale. The relationships among self- identity, PI/EA, and IA were examined using structural equation modeling. The severity of self-identity confusion was positively associated with both the severity of PI/EA and the severity of IA. In addition, the severity of PI/EA indicators was positively associated with the severity of IA. These results demonstrated that the severity of self-identity confusion was related to the severity of IA, either directly or indirectly. The indirect relationship was mediated by the severity of PI/EA. Self-identity confusion and PI/EA should be taken into consideration by the community of professionals working on IA. Early detection and intervention of self-identity confusion and PI/EA should be the objectives for programs aiming to lower the risk of IA. Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) and impulse control disorder in the DSM-IV Text Revision [Internet addiction (IA) has become a major global public health problem. Several studies have proposed diagnostic criteria for IA based on various concepts originally adopted from formal psychiatric disorders such as substance dependence and pathRevision . AlthougRevision and the Revision ,6,7. TheIA is predominant among young people, especially college students. College students normally have free and unlimited access to the Internet, flexible schedules, and are free from their parents\u2019 interference. They use the Internet for studying, gaming, social networking, gambling, chatting, shopping, and watching pornographic videos ,9. PreviInvestigating the factors related to IA is the first step toward preventing its incidence and providing early treatment to improve mental health. IA is the result of interaction between individuals and their environment ,19. SeveSelf-concept formation, also termed personal identity formation, as a main developmental task in adolescence, includes the acceptance of physical changes and the development of social and emotional competencies and self-efficacy . PreviouPsychological inflexibility and experiential avoidance (PI/EA) are defined as rigid strategies guided by psychological reactions and an unwillingness to experience unpleasant events or privations . PI/EA cResearch has determined three common types of self-identity confusion: Disturbed identity , unconsoNo study has examined the relationship between self-identify confusion, PI/EA, and IA. This study investigated the mediating effects of PI/EA on the relationship between self-identity confusion and IA among college students. We hypothesized that self-identity confusion is positively associated with IA and that PI/EA mediate the relationship between self-identity confusion and IA.Participants were recruited using an advertisement posted for college students aged between 20 and 30 years. Five hundred and three college students responded to the advertisement. Of them, three were excluded due to difficulties in understanding the meaning of research questionnaires. In total, 238 male and 262 female college students coming from 70 colleges located across Taiwan participated in this study. Their mean age was 22.1 years, with a standard deviation of 1.8 years. Written informed consent was obtained from all the participants prior to the assessment. The study was approved by the Institutional Review Board of Kaohsiung Medical University Hospital.Self-Concept and Identity Measure. We used the Self-Concept and Identity Measure (SCIM) to assess the level of self-identity confusion [onfusion . The 27-onfusion . Each itChen Internet Addiction Scale. We used the self-administered Chen Internet Addiction Scale (CIAS) to evaluate the participants\u2019 severity of IA in the month preceding the study. The CIAS contains 26 items that are rated using a four-point Likert-type scale, with the total score ranging from 26 to 104 [6 to 104 . A highe6 to 104 .The Acceptance and Action Questionnaire-II. The Acceptance and Action Questionnaire-II (AAQ-II) [(AAQ-II) was revi(AAQ-II) . The AAQ(AAQ-II) . The CroResearch assistants explained the procedures and methods for completing the research questionnaires to the participants individually. The participants were allowed to ask questions if they encountered problems while completing the questionnaires, and the research assistants helped them to resolve their problems.The hypothesized model for the relationships among self-identity confusion, PI/EA, and IA is presented in The study procedures were carried out in accordance with the Declaration of Helsinki. The Institutional Review Boards of Kaohsiung Medical University Hospital approved the study . All participants were informed about the study and provided written informed consent. In total, 85 (17%) participants were identified as having Internet addiction. The means, standard deviations, and correlation matrices of the measured variables are shown in p < 0.05).The goodness-of-fit indices of SEM for the hypothesized model on the relationships among self-identity confusion, PI/EA, and IA are listed in Moreover, all paths in the hypothesized model were significant. The severity of self-identity confusion was positively associated with the severity of IA and PI/EA. In addition, the severity of PI/EA was positively associated with the severity of IA. The severity of self-identity confusion was directly related to the severity of IA and indirectly related to the severity of IA through the increasing severity of PI/EA.This was the first study to examine the association between self-identity confusion, PI/EA, and IA. The results of this study revealed that self-identity confusion was directly related to IA and indirectly related to IA by the mediation of PI/EA. The results of previous studies on the relationship between self-identity styles and IA have been mixed. Some studies have demonstrated that normative styles of self-identity are protective factors of IA, whereas diffuse-avoidant styles are the risk factors of IA ,54. SeveThe present study discovered that PI/EA were significantly associated with IA in college students. The relationship between PI and IA has also been supported by the results of an imaging study that suggested that people with IA had difficulties in executive control and attention when switching tasks . People In this study, we found that PI/EA mediated the relationship between self-identify confusion and IA. A few possible mechanisms may account for the result. First, an unsuccessful formation of self-concept may lead individuals to experience self-identity diffusion, thereby increasing the risk of mental disorders, such as personality, depressive, and addictive disorders. These disorders usually persist into adulthood if they are not treated appropriately ,63 and cSecond, people may experience various social-cognitive processes to construct and maintain their self-identity. Beronsky (1992) proposed three processing orientations that contribute to form and maintain a sense of self-identity: A procrastinating, diffuse-avoidant style; an open, informational style; and a conforming, normative style . The indOn the basis of our study results, we suggest that school counselors and mental health professionals routinely evaluate whether college students with IA have self-identity confusion and high PI/EA. For self-identity confusion, interventions that provide relational safety and support self-validity and self-exploration may be helpful in reducing the risk of IA .The development of IA should be considered on the basis of ecological systems concepts . Self-idOur study had a number of limitations. First, the cross-sectional research design limited our ability to draw conclusions regarding the causal relationship among self-identity confusion, PI/EA, and IA. Second, the study data were exclusively self-reported and may have therefore suffered from shared-method variance. Third, the participants in this study were college students who responded to the recruitment advertisement. Therefore, the results of this study might not be generalized to college students who did not participate in this study. Fourth, the present study used the AAQ-II to measure PI/EA. Although research determined that AAQ-II has adequate internal consistency and convergent and divergent validity , the iteDespite these limitations, this study contributes to the literature, as it is the first study to examine the relationship between self-identity confusion and IA and the mediating effects of PI/EA. Further research replicating this SEM model across different samples is suggested.On the basis of our study, we found that self-identity confusion was directly related to IA and indirectly related to IA by the mediation of PI/EA. Therefore, self-identity confusion and PI/EA should be taken into consideration by the professionals working on prevention of IA. Moreover, we proposed that early detection and intervention for self-identity confusion may help college students to develop and consolidate self-identity, reduce PI/EA, and lower the risk of IA."} +{"text": "This issue begins with a 50-state survey of current policy affecting occupational therapy (OT) and physical therapy (PT). Authors Bierman, Kwong, and Calouro of The Center for Connected Health Policy presents methodology and analyses worthy of emulation by other healthcare professions. Regulatory-based professional uniformity within and across states are worthy attributional goals, both to facilitate the use of single-profession based telerehabilitation and to support team-based inter-professional practice.The five articles that follow address the clinical applications of telerehabilitation. Each examines the contributions of telerehabilitation in a unique clinical circumstance. Two of five articles explore providers\u2019 experience and perspectives. The evaluation of the benefits of telerehabilitation to avoid missed appointments will resonate across most healthcare professions. Finally, the use of telerehabilitation to address chronic spinal pain has great promise to ameliorate suffering and avoid more aggressive physical management.IJT is grateful to new and returning reviewers; William E. Janes, OTD, MSCI, OTR/L, Section Editor; colleagues at the Rehabilitation Engineering Research Center on Information and Communication Technology Access at the University of Pittsburgh; and, the IJT publishers at the University Library System at the University of Pittsburgh, especially Vanessa Gabler, Electronic Publications Manager.We cordially invite submissions to the Spring 2019 issue by mid-February 2019. IJT accepts original research, case studies, viewpoints, technology reviews, book reviews, and country reports that detail the status of telerehabilitation.Sincerely,Ellen R. Cohn, PhD, CCC-SLP, ASHA-F, IJT EditorJana Cason, DHSc, OTR/L, FAOTA, Senior Associate Editor"} +{"text": "Alcohol use disorder (AUD) is a chronic, relapsing brain disease characterized by a reduced ability to stop or control alcohol use despite negative social, work, or health consequences. Often, it co-occurs and interacts with post-traumatic stress disorder (PTSD), which may develop after experiencing or witnessing a life-threatening event, such as combat, a natural disaster, a car accident, or sexual assault, and can result in shock, confusion, anger, and anxiety.Co-occurring AUD and PTSD is a public health concern, especially among active military service members and veterans, as well as victims of violence and sexual assault. Approximately one in three people who have experienced PTSD have also experienced AUD at some point in their lives.Alcohol Research: Current Reviews examines the current literature on the prevalence, diagnoses, causes, and risk factors of AUD and PTSD, their co-occurrence, and treatment for individuals facing both conditions.This issue of The Epidemiology of Post-Traumatic Stress Disorder and Alcohol Use Disorder, describe the changes in the Diagnostic and Statistical Manual of Mental Disorders (DSM) definitions of AUD and PTSD. They review key surveys that have measured these disorders, the possible relationships between the two disorders, the risk factors, and which populations are at risk.Smith and Cottler, in Functional and Psychiatric Correlates of Comorbid Post-Traumatic Stress Disorder and Alcohol Use Disorder, Straus and colleagues present the DSM-5 definitions for PTSD and AUD and discuss models for functional relationships between the disorders. They also examine risk factors and their associations with co-occurring disorders.In Common Biological Mechanisms of Alcohol Use Disorder and Post-Traumatic Stress Disorder, review animal models for and clinical studies of AUD and PTSD. They discuss the relevant neurobiological circuits and examine the role of stress in these disorders.Suh and Ressler, in Early Life Stress as a Predictor of Co-Occurring Alcohol Use Disorder and Post-Traumatic Stress Disorder. They review both human and preclinical models of these disorders and examine potential biologic, genetic, and epigenetic mechanisms.Lee and colleagues investigate childhood stress as a predictor for PTSD and AUD in Co-Occurring Post-Traumatic Stress Disorder and Alcohol Use Disorder in U.S. Military and Veteran Populations, Dworkin and colleagues report on the frequency of co-occurring PTSD and AUD in military personnel and veterans, and they examine population-specific factors contributing to the development of PTSD and AUD. They also describe evidence-based psychological and pharmacological treatments for these populations and suggest future directions for research on treatment effectiveness.In Alcohol Use Disorder and Traumatic Brain Injury. The potential neuropsychological and neurobiological mechanisms underlying those relationships are discussed.Weil and colleagues provide an overview of the bidirectional relationships between traumatic brain injury and AUD in Behavioral Treatments for Alcohol Use Disorder and Post-Traumatic Stress Disorder, Flanagan and colleagues describe evidence-supported behavioral interventions for treating AUD, PTSD, and co-occurring AUD and PTSD. They also examine the debate regarding sequential versus integrated treatment models.In Pharmacotherapy for Co-Occurring Alcohol Use Disorder and Post-Traumatic Stress Disorder: Targeting the Opioidergic, Noradrenergic, Serotonergic, and GABAergic/Glutamatergic Systems, Verplaetse and colleagues report on pharmacotherapies for co-occurring AUD and PTSD. They discuss current clinical trials for medications and highlight future directions for neurobiological targets that have potential for treating individuals with this dual diagnosis.In"} +{"text": "Introduction: Autonomic nervous system (ANS) dysfunction contributes to several non-motor symptoms of Parkinson's disease (PD). In addition, ANS plays a role in the genesis and maintenance of atrial fibrillation (AF). This study investigated the temporal association between PD and AF.Methods: Data were obtained from the National Health Insurance Research Database of Taiwan. In total, 15,375 patients with newly diagnosed PD were matched with four controls each based on the propensity score. This study was bidirectional. A case-control study for the odds ratio (OR) of AF before PD and within 2 years of PD diagnosis was evaluated through conditional logistic regression. Furthermore, a cohort study on the subdistribution hazard ratio (SHR) for new-onset AF 2 years after PD diagnosis was evaluated using competing risk analysis.Results: In the case-control study, PD was found to be significantly comorbid with AF . Subgroup analysis demonstrated that this association consistently presented in the absence of confounding factors of AF. In the cohort study, people with PD were found to have a lower risk of AF . However, a consistent association was not observed between the confounding factors of AF and PD during the subgroup analysis.Conclusions: This study demonstrated that the premotor and early stages of PD were comorbid with AF, whereas the risk of AF was lower in the later stages. Thus, AF might be a premotor predictive biomarker and comorbidity of early PD. Tremor, bradykinesia, rigidity, and postural instability are the cardinal motor symptoms of Parkinson's disease (PD). However, numerous non-motor symptoms (NMSs), such as depression, dementia, rapid eye movement sleep behavior disorder (RBD), and anosmia, are also comorbid with PD . The bioDegeneration of the autonomic nervous system (ANS) contributes to certain NMSs of PD, the best-known of which is constipation. More than 60% of people with PD develop constipation because of poor intestinal peristalsis caused by a dysfunctional vagus nerve. Moreover, in most cases, constipation heralds the onset of motor symptoms . This seCardiac rhythm is regulated by ANS as well. Sympathetic and parasympathetic innervations originate from the paravertebral ganglia and vagal nerves, respectively. Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia, and it is strongly associated with morbidity, mortality, and poor quality of life. AF stems from several etiologies, and rather than ischemic heart disease, heart failure, and hyperthyroidism, ANS plays a crucial role in AF, particularly for patients with no structural heart disease , 8.Considering the role of the vagal nerve\u2013related ANS system in PD and AF, PD may be comorbid with AF. Moreover, similar to other autonomic NMSs, AF may be a biomarker for the onset of PD motor symptoms. This study employed the National Health Insurance Research Database (NHIRD) of Taiwan to investigate whether AF is associated with newly diagnosed PD and evaluated the temporal relationship between both conditions.This study was approved by the Joint Institutional Review Board of Taipei Medical University (TMU-JIRB No. 201701058).This study was conducted using the NHIRD data files maintained by the Health and Welfare Data Science Center (HWDC). The NHIRD is a claims-based database managed by the National Health Insurance Administration of Taiwan; Taiwan's NHI provides coverage for 99% of its residents. The NHIRD files include inpatient, outpatient, and pharmaceutical claims and disease diagnoses coded according to the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM). In addition, the enrollment files of beneficiaries and providers were also included. The data in this study were from 2000 to 2015. Additionally, we linked the collected data with the national death registry to obtain death records. The two data sets can be linked according to the regulations of the HWDC. Both case-control and cohort (newly diagnosed AF 2 years after first PD diagnosis) studies were applied to examine the temporal relationship between PD and AF.Newly diagnosed people with PD were defined as those who had at least two diagnostic claims (ICD-9-CM: 332.0) and prescription claims for dopaminergic agents between 2004 and 2011. The index date of PD was defined as the date of first PD diagnosis, hereafter referred to as the index PD. People who were aged < 45 years, had a history of stroke, or had received any antipsychotic drug before the index PD, were excluded to avoid the possibility of misclassification of secondary Parkinsonism. In addition, several predisposing factors may trigger AF directly are widely utilized to diminish the confounding factors that inevitably occurs in studies investigating the effect of the exposures on the outcome. In PSM, matched are formed by virtue of sharing similar propensity score values. The weighted-value reveals the risk of the participant for the outcome of interest according to their underlying characteristics predispose them for that outcome irrespective of the exposure of interest .In this study, the propensity score was measured on the basis of hypertension , diabetes mellitus , hyperlipidemia (ICD-9-CM: 272), chronic heart failure , coronary artery disease , chronic lung disease , renal disease (ICD-9-CM: 580\u2013589), and inflammatory dx . The selection of these factors was based on the association with AF genesis . ControlBoth the people with PD and controls were tracked or followed up for their risk of AF according to the study design. Patients with AF were defined as those who first had at least two diagnostic claims (ICD-9-CM: 427.31) and prescription claims for warfarin or oral anticoagulant agents without claims for venous thrombosis (ICD-9-CM: 453), pulmonary embolism , and antiphospholipid syndrome (ICD-9-CM: 286.53 and 286.59). AF, especially paroxysmal AF, is usually under-diagnosed with a certain latent period before diagnosis. Hence, in the case-control study, AF risk was measured within 2 years of PD or before the index date of PD/pseudo-PD diagnosis. In the cohort study, AF risk was measured 2 years after the index date of PD/pseudo-PD diagnosis. The selection process is presented in P < 0.05 was considered significant.Baseline characteristics were analyzed using standardized mean difference (SMD). SMD > 0.1 indicated non-negligible differences between the two groups. The case-control study for the odds ratio (OR) of AF before PD and within the 2-year interval of PD diagnosis was evaluated using conditional logistic regression, and a cohort study for the subdistribution hazard ratio (SHR) of new-onset AF after PD diagnosis was evaluated using competing risk analysis. Because the participants were at a high risk of mortality, we applied competing risk model analyses to estimate the absolute relative AF risks. The follow-up period for each patient ranged from the index date of PD/pseudo-PD diagnosis to the date of AF diagnosis or death or the end of the observation period . All analyses were performed using SAS/STAT version 9.4 and STATA 14 . A Initially, the study included 15,434 newly diagnosed people with PD, among whom 59.2% were men. After the 1 to 4 PSM, 15,375 subjects remained in the study. After PSM, their mean age was 71.7 \u00b1 9.9 years. The prevalence of previous or current comorbidity was 47.8, 20.7, and 2.5% for HTN, DM, and CHF, respectively .In the case-control study, compared with matched controls, newly diagnosed people with PD were significantly comorbid with AF . AccordiIn the cohort study, newly diagnosed people with PD had lower AF risk during the follow-up period . In the This study demonstrated that people with PD were more likely to be comorbid with AF before and during the onset of motor symptoms. By contrast, people with PD were at a lower risk of AF in the later stages of PD. This discrepancy in the temporal relationship between the two diseases indicates that, similar to other ANS symptoms, AF may be an early NMS of PD.ANS dysfunction is a common feature in the premotor and early stages of PD . For exaANS plays an essential role in the genesis and maintenance of AF, and both the sympathetic and parasympathetic nerves regulate cardiac rhythm through the paravertebral sympathetic ganglia and vagal nerve, respectively . Unlike Using a nationwide population-based method, this study demonstrated the association between AF and newly diagnosed PD, finding that newly diagnosed people with PD were more likely to be comorbid with AF before and during the onset of motor symptoms. This association and temporal relationship support the hypothesis of caudal\u2013rostral spreading of \u03b1-synuclein pathology, which indicates that the medullary vagal nucleus is the first region in the CNS to be involved in such pathology, followed by the onset of motor symptoms parallel to \u03b1-synuclein accumulation in the midbrain. Conversely, they were at lower risk of AF during the follow-up period. After the diagnosis of PD, people tend to have a healthier lifestyle and regular medical check on the blood pressure, blood glucose, and lipid profile, which resulted in better general health condition than population and contribute to the reduction of the risk of AF. It was a limitation of NHIRD study that we could only match by the presence of disease claim without awareness of the severity of the disease. Meanwhile, in most of the occasion, AF is only detected when it became symptomatic, such as tachycardia or leading to stroke. People with PD, especially those with prominent tremor, may prescribe beta-blockers for controlling the tremor, which may mask the AF-induced tachycardia and under-diagnosed the AF.A notable contribution of this study is identification of the temporal association between AF and PD. Although the prevalence of AF and PD is low, they remain major public health concerns. Moreover, as an ANS-related NMS of premotor PD, AF can serve as one of the predictive biomarkers of PD in the same manner as RBD, anosmia, and constipation, thereby improving the prediction accuracy of PD onset. For further future study to identify the association between AF with PD in the pre-motor stage, it is suggested that for people at high risk of PD (such as RBD or anosmia), the wearable applications should be considered to identify the AF or other kinds of cardiac arrhythmia in a longer-term, more comprehensive manner.However, this study has several limitations. AF in some patients may be paroxysmal and asymptomatic, which results in delayed diagnosis. Thus, the temporal relationship between AF and PD may be biased. People with early motor symptoms of PD may visit clinics more frequently and receive more examinations, which increases the possibility of asymptomatic AF diagnosis. To eliminate this bias, this study adjusted the potential effect of the frequency of clinic visits when examining the association between AF and PD. In addition, many confounding factors occur in AF genesis, such as conventional vascular risk factors, structural heart disease, and systemic illnesses. This study did not investigate lone AF, which is the most ANS-related AF, because patients with lone AF are rare even in the NHRID database. Nevertheless, a subgroup analysis was performed that found that the association between PD and AF was significant in the absence of any one of them. Therefore, minimizing the confounding factors of AF may strengthen association between the two diseases. Another limitation is that although PD diagnosis has been validated with satisfactory accuracy, there is a possibility of false classification of AF and PD in claims-based medical database research . MoreoveIn conclusion, this study revealed that AF may be an NMS of PD, and people with PD were comorbid with AF before and during the onset of motor symptoms. Future studies may include AF as a premotor biomarker to develop a comprehensive PD prediction model.(A) Conception: all. Organization: C-TH, LC, and L-NC. Execution: C-TH, LC, DW, and L-NC.Research project: (A) Design: C-TH, DW, and W-TC. (B) Execution: W-TC and L-NC. (C) Review and Critique: all.Statistical Analysis: (A) Writing of the first draft: C-TH, LC, and DW. (B) Review and Critique: all.Manuscript: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."} +{"text": "OBJECTIVES/SPECIFIC AIMS: Responding to the need and interest of students and faculty of the UHSP in learning about CTR, the Title V Cooperative Project between UPR-MSC and UCC, developed and offered a training cycle (TC) in CTR. METHODS/STUDY POPULATION: Undergraduate students (US), undergraduate faculty (UF), and graduate students (GS) were invited to register in: Research Education Towards Opportunities (RETO) and Mentorship Offering Training Opportunities for Research (MOTOR), which consisted of 20 hours of training in CTR, with interdisciplinary sessions in: Introduction and preparation of a presentation in CTR; Identify, interview and share a presentation of a CT researcher; participation in conferences and a summer camp in CTR. At the end of the TC, surveys\u2014satisfaction and needs assessment\u2014for training in CTR were administered. RESULTS/ANTICIPATED RESULTS: Thirty-three (33) registered in the TC, distributed: 13 (39.39%) US in RETO, 12 (36.36%) GS and 8 (24.24%) UF in MOTOR. Of these, 25 (75.75%) answered and submitted the on-line surveys and received a completion certificate. All (100%) were satisfied with the TC, and for 96% of the respondents, their expectations were fulfilled, and will continue in the TC. They selected critical review, scientific communication, and cultural diversity as thematic areas of interest. In addition, 60% of them selected neuroscience, cancer and medical imaging as main research areas of interest. DISCUSSION/SIGNIFICANCE OF IMPACT: The TC demonstrated to be an effective strategy to provide new knowledge, experiences, and interest in CTR. It also established a pathway for future engagement in CTR."} +{"text": "Dr. Francois Niyonsaba is not included in the author byline. He should be listed as the tenth author, and his affiliation is the Atopy Research Center, Juntendo University Graduate School of Medicine, Tokyo, Japan. The contributions of this author are as follows: Conceptualization and resources.The following information is missing from the Funding section: This study was supported by Grant-in-Aid for Young Scientists (B) (16K19855) from the MEXT, Japan to KS."} +{"text": "Scientific Reports 10.1038/s41598-018-37033-4, published online 28 January 2019Correction to: The original version of this Article contained errors in the spelling of the authors Madison S. Cox, Andrew J. Steinberger & Joseph H. Skarlupka which were incorrectly given as Madison Cox, Andrew Steinberger & Joseph Skarlupka respectively.Additionally, the original version of this Article contained an error in Affiliation 4, which was incorrectly given as \u2018Department of Microbiology, University of Wisconsin-Madison, Madison, WI, 53706, USA\u2019. The correct affiliation is listed below:Department of Bacteriology, University of Wisconsin-Madison, Madison, WI, 53706, USAFinally, the Acknowledgements section in this Article was incomplete and contained typographical errors.\u201cWe thank Laura Cersoscimo and Rafael Oliveira for their generous help on calf tissue collection. WL and DB were supported by appropriated project 5090-31000-026-00-D from the USDA Agriculture Research Service (Dairy Forage Research Center). JW was supported by appropriated project 5090-43440-006-00-D from the USDA Agricultural Research Service . Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation by the US Department of Agriculture. The USDA is an equal opportunity provider and employer.\u201dnow reads:\u201cWe thank Laura Cersosimo and Rafael Oliveira for their generous help on calf tissue collection. WL and DB were supported by appropriated project 5090-31000-026-00-D from the USDA Agriculture Research Service (Dairy Forage Research Center). JW was supported by appropriated project 5090-43440-006-00-D from the USDA Agricultural Research Service . GS was supported by, in part, by a USDA National Institute of Food and Agriculture HATCH grant WIS02007 and a USDA Agriculture and Food Research Initiative Competitive Grant no. 2015-67015-23246. MSC was supported by a USDA Agriculture and Food Research Initiative Education and Literacy Initiative predoctoral fellowship no. 2018-67011-27997. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation by the US Department of Agriculture. The USDA is an equal opportunity provider and employer.\u201dThese errors have now been corrected in the PDF and HTML versions of the Article, and in the accompanying Supplementary Information file."} +{"text": "Nature Communications; 10.1038/s41467-018-07310-x; published online 19 November 2018Correction to: The original version of this Article contained errors in the author affiliations. Mehmet Fatih Bolukbasi was incorrectly associated with Bluebird Bio., Cambridge, MA, USA and Ankit Gupta was incorrectly associated with Exonics Therapeutics, Watertown, MA, USA. This has now been corrected in the HTML version of the Article. The PDF version of the Article was correct at the time of publication."} +{"text": "AbstractOpiliones fauna of Canada is reviewed and summarised. At present, 36 native and seven non-native species have been documented in Canada using traditional morphological taxonomy, although more than 20 species may remain undiscovered based on species diversity in the adjacent United States and evidence from DNA barcoding. Consequently, the native fauna is yet to be fully explored and the number, distribution and ecological effects of non-native species remain unclear. Until the 1960s, work on the Canadian Opiliones fauna was largely conducted by researchers based outside the country. From that time on, several Canadian workers became active. However, these taxonomists have now retired and no one has assumed their role. Thus, there is a need to invigorate taxonomic research on the harvestmen of Canada and for the production of easy-to-use identification tools for use by non-taxonomists.The taxonomic diversity of the Opiliones, or harvestmen, encompass over 6600 described species and about 50 families worldwide , 979, 1). https://bugguide.net), including first national records for Crosbycusdasycnemus (Crosby) by Leiobunumnigropalpi (Wood) by The rate of discovering and cataloguing Canadian harvestmen has been a slow and largely international process that has tended to lag behind that of other western countries. The year in which each native species was first recorded, data mined primarily from Opiliones are divided into four suborders with the following relationships: Cyphophthalmi, ). Thus, the current taxonomic hierarchy does not strictly reflect the generally accepted phylogeny. Laniatores, Dyspnoi, and Eupnoi form the clade Phalangida, which is the sister group to Cyphophthalmi, and the Dyspnoi and Eupnoi are united in the Palpatores, which is the sister group to Laniatores and S.acaroides (Ewing), with northern Washington populations of the latter now known as S.boyerae Giribet & Shear, 2010, might range into southern British Columbia. Global biogeoPageBreakPageBreakgraphic patterns indicate that Sironidae is the only family likely to occur in Canada (Erebomasterflavescens (Cope) (Cladonychiinae: Travuniidae) in southeastern New York, an observation that is often overlooked , just tw (Banks) . Bishop ed e.g., . Given tDyspnoi contains three main lineages, Ischyropsalidoidea, Troguloidea, and Acropsopilionidae, with the latter recently transferred from Caddoidea (Eupnoi) , with Sabacon species being widespread in Canada and with barcoding data suggesting the existence of greater species diversity than current taxonomy would suggest and to the subfamily Ceratolasmatinae within the family Ischyropsalididae, which otherwise contains only the European Ischyropsalis. Among the four families in Troguloidea, only the Nemastomatidae occur in Canada, specifically the native ortholasmatines, Dendrolasma and Ortholasma, in British Columbia (Nemastomabimaculatum (Fabricius), in the East (Acropsopilioboopis (Crosby), is known from Canada and no additional species are expected to occur there.The morphologically diverse Holarctic suborder (Eupnoi) . The famnisation . Sabaconst Table . The CerColumbia , and thethe East . Only onEupnoi consists of two superfamilies, the species-poor Caddoidea, and the species-rich Phalangioidea. Caddoidea s. str. , Sclerosomatidae (36% of known Canadian species), and at least one species of Protolophidae . Six phalangiid species are native to Canada: Mitopusmorio (Fabricius), Odielluspictus (Wood), Leptobunusborealis Banks, Leptobunusparvulus (Banks), Liopilioyukon Cokendolpher, and PageBreakLiopilioglaber Schenkel, and at least six appear to have been introduced from Europe: Oligolophustridens (CL Koch), Paroligolophusagrestis (Meade), Opilioparietinus (DeGeer), Phalangiumopilio Linneaus, Lophopiliopalpinalis (Herbst), and Rilaenatriangularis (Herbst) (Sclerosomatidae are represented by five native genera: Hadrobunus (2 spp.), Leiobunum (9 spp.), Leuronychus (1 sp.), Nelima (2 spp.), and Togwoteeus (1 sp.).The suborder s. str. containsisconsin . The famAcuclavella in Eumesosomaroeweri (Goodnight and Goodnight) and Trachyrhinus [T.favosus (Wood) and/or T.marmoratus Banks] (see Cokendolpher 1981) to be present in counties along the USA-Canadian border. This would add two genera and one subfamily (Gagrellinae) to the Canadian fauna. Canada also seems prone to the introduction and establishment of European harvestmen, especially phalangiids (Trogulustricarinatus (Linneaus) in New York and Massachusetts has revealed two species in Ontario (J Shultz unpubl. data), where only one was assumed to exist. On the other hand, in Leiobunum (Sclerosomatidae), L.formosum (Wood) and L.nigripes Weed were found to be junior synonyms of L.verrucosum (Wood) , which em (Wood) , may indLeiobunumvittatum (BOLD:AAM8191) contains no specimens of that species but encompasses at least three morphologically and geographically distinct species of Hadrobunus. Lastly, specimens in many BINs are identified to genus PageBreakor species when, in fact, the photos show either juveniles or otherwise unidentifiable specimens. In some cases, the ambiguous specimens are determined as European adventives that are otherwise unrecorded from Canada, a situation that, if correct, could suggest an early stage in the expansion of a potential invasive species. In fact, results from barcoding based on accurate identifications (BOLD:AAM8194) revealed a previously unknown introduction and expansion of the European Oligolophustridens in Alberta, Saskatchewan and extreme southeastern British Columbia. Clearly, surveys of the harvestman fauna should be undertaken throughout the country to establish the species composition of the native fauna as well as the distribution and environmental impacts of the comparatively numerous species that have been introduced into Canada.Progress in the discovery and understanding of harvestman diversity in Canada will require the effort of one or more professional taxonomists to engage in active, modern research on the order. A particularly urgent goal is the production and distribution of accessible and easy-to-use tools for the identification of the harvestmen species known or likely to occur in Canada. The virtual absence of such resources has already had significant negative consequences. For example, of the 64 BINs of Canadian harvestmen in the Barcodes of Life Data systems (BOLD) , about 2"} +{"text": "In the one-to-one propensity-score matched cohort (317 neurologists versus 317 neurosurgeons), there were even higher risks among neurosurgeons than neurologists . Neurosurgeons have a higher chance of intervertebral disc disorders than neurologists. This is potentially an occupational risk that warrants further investigation.High physical activity or workload has been associated with intervertebral disc degeneration. However, there is little data on physicians\u2019 risks of disc disease. The study aimed to investigate the incidences of spinal problems among neurologists and neurosurgeons. A cohort of neurologists and neurosurgeons was derived from Taiwan\u2019s national research database. During the study period, the incidences of intervertebral disc herniation or spondylosis among these specialists were calculated. Another one-to-one by propensity score matched cohort, composed of neurologists and neurosurgeons, was also analyzed. A Cox regression hazard ratio (HR) model and Kaplan-Meier analysis were conducted to compare the risks and incidences. The entire cohort comprised 481 and 317 newly board-certified neurologists and neurosurgeons, respectively. During the 15 years of follow-up, neurosurgeons were approximately six-fold more likely to develop disc problems than neurologists (crude HR = 5.98 and adjusted HR = 6.08, both Herniation of intervertebral discs could cause back pain, radicular pain, or even muscle weakness. These symptoms have been demonstrated to cause a lower quality of life in patients when compared with healthy populations ,2. For eThe present study aimed to investigate the incidences of intervertebral disc herniation among professional medical doctors treating neurological disorders, who frequently encounter patients with the same problem. Furthermore, the study aimed to address the discrepancy in risks among neurologists and neurosurgeons, who treat the disorder differently . The present study uniquely acquired a cohort of physicians and surgeons over 15 years, using the National Health Insurance Research Database (NHIRD) of Taiwan. Due to the comprehensive and monopolistic coverage of Taiwan\u2019s national health insurance , this coThe study used the NHIRD, which contains comprehensive information on every health care worker and insured subject of Taiwan, provided by the National Health Research Institute for academic use. The NHIRD allows detailed information of every individual (coded for deidentification), including gender, date of birth, date of medical board certification, residential or work area, dates of clinical visits, hospitalization, diagnoses, procedures, prescriptions, and expenditure amounts, etc. The diagnoses and procedures were recorded according to the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) codes and were originally used for billing. Thus, these data were internally monitored and cross-checked. Furthermore, the government-operated health insurance program, National Health Insurance (NHI) of Taiwan, has enrolled 99% of the population and provides unrestricted access to medical care in contracted clinics and institutions, which comprise 97% of the providers of health care services in Taiwan. Therefore, the NHIRD uniquely collected all data of patients, physicians, and physician-patients, and the cohort study had an extremely high rate of follow-up, theoretically almost complete follow-up of these physicians.This study was approved by the Institutional Review Board of Taipei Veterans\u2019 General Hospital, Taiwan (VGHIRB No.: 2012-10-008BC) and National Yang-Ming University Hospital (IRB No. 2015A015). Written informed consent from each of the enrolled was waived because all identifying personal information in NHIRD was stripped before analysis.The study contained two cohorts, the original cohort, and the matched-cohort; both were extracted from the NHRID by identifying board-certified medical specialists.In the original cohort, every newly certified neurologist and neurosurgeon was identified and included for follow-up. Exclusion criteria were previous history of spinal problems before the board-certification, unregistered physicians who had not been in practice in any of the hospitals, or whose certification was suspended.The other matched cohort, a one-to-one matched comparison among neurologists and neurosurgeons, was derived from the original cohort based on a propensity score calculated by sex, level of hospital in which they were working, and in-patient service volume. The matched cohort, therefore, shared the same inclusion/exclusion criteria with the original cohort. Sex, age, hospital level, and service volume were considered covariates of the study.Every neurologist and neurosurgeon was subsequently followed-up after their board certification. During the observation period, any hospitalization coded with the ICD-9 diagnosis of spinal intervertebral disc diseases was extracted and deemed as the study end-point. This cohort only censored follow-ups in the following conditions: the subject stopped practice , expired, the date of the end-point incidence, or the end of this study, 31 December 2013 .http://www.jct.org.tw/FrontStage/doha_en.html), a government-sponsored organization, and classified into three levels: medical center, regional hospital, and district hospital. The highest level of hospitals, i.e., medical centers, in Taiwan, hold the highest standard in academics, quality of care, and responsibility of public health. Although the public might consider that the higher accreditation implies a better quality of medical services, the severity of diseases of the patients does not necessarily correlate with the classification of hospitals. However, the level of hospital, indeed, reflects some working styles of these medical specialists. For example, a larger proportion of medical centers are affiliated with medical schools and employ more university faculties. On the other hand, all these medical centers, regional hospitals, and district hospitals are contracted with the NHI program, which covers almost the entire population of Taiwan. Therefore, all the doctors, doctor-patient, and end-point events of the study were included in this cohort, no matter which hospital they went to or in which they worked in.All the medical specialists in Taiwan, including physicians and surgeons, are only allowed to practice in one hospital (to which they are registered). In the present study, each of the neurologists and neurosurgeons in Taiwan was assigned to the hospital where he or she received their board certification. These hospitals of Taiwan are evaluated regularly for hospital accreditation by the Joint Commission of Taiwan and analyzed by the SPSS software . The cumulative incidence rates of the study\u2019s outcome were estimated and compared using the Kaplan-Meier method and Log-rank test. The Cox proportional hazard model was used to compare the hazard ratios of such outcomes among neurologists and neurosurgeons after adjustment for the covariates, including sex, age, hospital level, and service volume. A two-tailed level of 0.05 was considered statistically significant.n = 2), not registered to practice (n = 27), or who never worked in a hospital (n = 97), there were 798 medical specialists in the original cohort.Since 1 January 1998, there were a total of 924 newly board-certified neurologists and neurosurgeons identified in the NHIRD. After exclusion of those who had a history of spinal diseases (p < 0.001). The neurologists and neurosurgeons also had a different composition of the hospital level they worked in, and their volume of in-patient services (both p < 0.001) differed. The overall incidence rate of spinal disc diseases requiring hospitalization was 1.76 per 1000 person-years (p < 0.001). After adjustments made for age, sex, hospital level, and service volume, the neurosurgeons were six-fold more likely to have such problems . The neurosurgeons were significantly more male predominant and older than the neurologists (both on-years . Neurosu < 0.05) .p < 0.001 and p < 0.05, respectively) (In the other matched (one-to-one by propensity score) cohort, composed of 317 neurologists and 317 neurosurgeons, the composition of gender, level of working hospital, and in-patient service volume were very similar, except the neurosurgeons were approximately 2.4 years older at the time of board-certification than the neurologists . The risctively) .p < 0.05) during the 15 years the differences in the outcome event demonstrated among neurologists and neurosurgeons warrant further attention. This is potentially a risk that might adversely affect a specialist\u2019s career and could be avoided or at least reduced.Higher incidences of intervertebral disc disorders were demonstrated in neurosurgeons than neurologists. Whether this is potentially an occupational risk warrants further investigation."} +{"text": "Following publication of the original article , the autAddition to the \u2018Methods\u2019 section: The seeds for 41 of the populations grown in the common garden experiment were provided by the research group of Prof. Ali-Shtayeh, Biodiversity and Environmental Research Center, Til, Nablus, Palestine, as part of MERC-USAID collaborative project TA-MOU-12- M32\u2013016. The above 41 populations were described by Ali-Shtayeh et al. . Data for each accession were also provided to Perl-Treves, along with a soil sample from each field. Part of the fruit pictures included in Fig. 1A was provided as well.The authors thank Mr. Nabil Omari of the Extension services of the Israeli Ministry of agriculture for agronomic advice and Mr. David Levy for technical assistance.Revised \u2018Availability of data and materials\u2019 section: Full phenotypic and genotypic data files that served to generate this study are deposited in the Israeli Genbank database https://igb.agri.gov.il/ and linked to the snake melon collection described in this study . Seed samples for research purpose will be available from the Israeli Gene Bank."} +{"text": "The biology of aging is an immensely complex process that impacts function at the sub-cellular, cellular, tissue, organ, and whole organism levels. New technologies and systems-level approaches provide an opportunity to greatly enhance our understanding of this complexity."} +{"text": "The Pneumonia Etiology Research for Child Health (PERCH) Study Group. Causes of severe pneumonia requiring hospital admission in children without HIV infection from Africa and Asia: the PERCH multi-country case-control study. Lancet 2019;394:757\u201379\u2014In this Article, the list of contributors to the study design was incorrect. David R Murdoch contributed to designing the study, not David P Moore. This correction has been made to the online version as of Aug 29, 2019, and the printed version is correct."} diff --git a/PMC_clustering_80.jsonl b/PMC_clustering_80.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..ef92fde067374da51aba814c490ae3f098f9ca81 --- /dev/null +++ b/PMC_clustering_80.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:2fc6fa3c90735a032d7866a1840ac78f532d50c70451577cd6e4ebe5cbddefd2 +size 45308813 diff --git a/PMC_clustering_81.jsonl b/PMC_clustering_81.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..8ade3f21fdb3bfb60a430ad7ed8129ab3e6c1473 --- /dev/null +++ b/PMC_clustering_81.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:8ea423501229ef7fd0117327c80b2f8ed579202bd30149a92829815d21d8ee77 +size 75371489 diff --git a/PMC_clustering_82.jsonl b/PMC_clustering_82.jsonl new file mode 100644 index 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--- /dev/null +++ b/PMC_clustering_94.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:44e8642e1763273a4fd78ce338dae47feefb169f8ed40ce161884ed78a1bafb2 +size 94861489 diff --git a/PMC_clustering_95.jsonl b/PMC_clustering_95.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..2666b63527a309acab35da9817ccc7fdacbd64e2 --- /dev/null +++ b/PMC_clustering_95.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:6ddde4e7e42d11599301a9beb7a899be131eded8130939d52d4189e609bf30fa +size 36948867 diff --git a/PMC_clustering_96.jsonl b/PMC_clustering_96.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..cde22d893760f4df5c707c5b66113af83c846793 --- /dev/null +++ b/PMC_clustering_96.jsonl @@ -0,0 +1,192 @@ +{"text": "In the editorial section, Philip Setel et al. (374) emphasize the need for rapid mortality surveillance during a pandemic. Jack Collins et al. (375) review ethical considerations of using mystery shoppers to study pharmaceutical sales.International Health Regulations. Jo\u00e3o Apr\u00edgio Guerra de Almeida speaks to Andr\u00e9ia Azevedo Soares (380\u2013381) about the origins of Brazil\u2019s human milk bank network and the psychosocial aspects of breastfeeding.In the news section, Lynn Eaton and Gary Humphreys (378\u2013379) report on calls to address the limitations of the Fernando C Wehrmeister, et al. (394\u2013405) estimate inequalities in coverage.Andr\u00e9 Loua et al. (420\u2013425) compare donation policies and programmes.Mireya Vilar-Compte et al. (382\u2013393) estimate costs of maternity leave to support breastfeeding.Catherine Jan et al. (435\u2013437) focus on prevention efforts.Maris Taubea and Wilm Quentin (426\u2013430) study the impact of community-based programmes.Shrey Mathur et al. (406\u2013412) compare five dosing guidelines.Ariadna Nebot Giralt et al. (413\u2013419) survey nongovernmental organizations. Igor Toskin et al. (431\u2013434) propose combining behavioural and biomedical interventions.Shah Ebrahim et al. (438\u2013440) argue for improvements to the indicator definition."} +{"text": "This month\u2019s theme issue is on the ethics of applying artificial intelligence to the health sector. In the editorial section, Kenneth Goodman et al. (230) introduce the World Health Organization\u2019s expert group on artificial intelligence in health and Bernardo Mariano (231) describes its global strategy on digital health. Flavia Bustreo and Marcel Tanner (232) imagine inclusive provision of health services in a digital age.\u00a0 In the news section, Gary Humphreys (235\u2013236) reports on health sector security risks and regulatory requirements for digital systems. He interviews Tze-Yun Leong (237\u2013238) about the engineering skills needed to develop artificial intelligence capable of dealing with real-life problems.\u00a0Maxwell J Smith et al. (290\u2013292) consider the digital divide, algorithmic biases, competing values and opaque decision-making. Alon Vaisman et al. (288\u2013289) review the ethical quandries of diagnostic imaging for neglected tropical diseases. Calvin W L Ho et al. (263\u2013269) describe features of a robust ethical and regulatory environment. Vijayaprasad Gopichandran et al. (277\u2013281) inspect the challenges of using demographic and biometric data to allocate services.Kristine B\u00e6r\u00f8e et al. (257\u2013187) analyse European Commission guidelines.Gabrielle Samuel & Gemma Derrick (239\u2013244) propose a process for ethics governance. Angeliki Kerasidou (245\u2013250) explores trade-offs in automated provision of care.Ibrahim Habli et al. (251\u2013256) consider the moral accountability of clinical decision-making systems.Amy K Paul & Merrick Schaefer (282\u2013284) suggest safeguards for national deployment of digital innovations. \u00a0Claire Louise Thompson & Heather May Morgan (293\u2013295) detail a code-of-conduct for artificial intelligence use in the national health service.Nicholas C Jacobson et al. (270\u2013276) explore ethical dilemmas posed by machine learning. David D Luxton (285\u2013287) takes a cautious look at conversational agents."} +{"text": "Ieder gesprek bevat tegenwoordig in een van de eerste zinnen het woord Corona. Zo ook dit editorial, met een hoofdletter geschreven vanwege de impact van het virus op alles wat we doen. Een tijdschrift heeft daar in eerste instantie minder last van. De vergaderingen van de editorial board gingen al telefonisch en correspondentie on line, Coronaproof. Een veel later effect is dat de hoeveelheid kopij zal afnemen; er worden minder pati\u00ebnten gezien en veel research ligt relatief stil. Gelukkig komen activiteiten in de urologische kliniek weer op gang, maar dat het anders zal worden, is inmiddels wel duidelijk.De artikelen in dit nummer gaan over zorg die een tijd stil heeft gelegen; ze werden ook al v\u00f3\u00f3r de Coronacrisis ter review aangeboden. Het artikel van Jacobs en collega\u2019s gaat over PROM/PREM\u2019s bij lithiasis. Een interessant onderzoeksthema over een onderwerp dat we vooral van Davinci-prostatectomie\u00ebn kennen. Mekke en collega\u2019s geven in het daarop volgende artikel het TURP/blaassteendilemma goed weer, waarna Jansen en collega\u2019s de rol van de testisprothese nog eens onder de aandacht brengen.Nog een gevolg van Corona: COVID-19 heeft ervoor gezorgd dat we elkaar niet fysiek ontmoeten tijdens het EAU-congres van dit jaar. Als het goed is, gaat dat echter wel in 2022 gebeuren, ook \u2013 net zoals nu de bedoeling was \u2013 in Amsterdam. Dus we krijgen als Nederlandse urologen n\u00f3g een keer de kans om dit event in ons land te hosten, alhoewel het de vraag is of dat weer met 15.000 deelnemers zal zijn. De EAU 2020 is dus online en alle abstract zullen ook langs die weg gereviewd worden volgens het junior/senior EAU-reviewprincipe. Het geeft ongetwijfeld weer een goed overzicht van de laatste ontwikkelingen op ons vakgebied, ontwikkelingen die nog niet door Corona zijn be\u00efnvloed.Dus, veel leesplezier, daar hebben we meer tijd voor, en blijf Coronavrij (en dat is zes)."} +{"text": "Dear Editor,et al. has shown ongoing trends for emigration and implicated several factors. While migration has always been a part of medical training, there is increasing concern about professionals leaving Ireland without intending to return . This worrying phenomenon has previously been attributed to budget cuts and deteriorating working conditions in the Irish Health System , family/friends time (43%), sleep (39%) and listening to music (33%). Less helpful mechanisms are also evident \u2013 isolation (41%), junk food (32%), alcohol (23%) or cannabis (1%) (West et al.et al.et al.Secondly, supportive organisational actions should be included in any hospital culture (Shanfield et al.et al.et al.et al.et al.Specific, evidenced interventions have utility on the ground and improve understanding and communication. Groups like \u2018Schwartz rounds\u2019 support hospital-wide communication sessions, foster positive interdisciplinary work and impact positively on staff satisfaction (Mayben et al.Although it is well and good to devise specific interventions, it should be stressed that under-resourcing remains a major factor for burnout (West To address health staff\u2019s issues, the Irish Health Service Executive created in April 2018 a new unit dedicated to staff welfare \u2018Workplace Health and Wellbeing Unit\u2019 (HSE Work Well Human Resource Services"} +{"text": "In the editorial section Haileyesus Getahun et al. (442) propose five measures to improve antimicrobial use and oversight during the COVID-19 pandemic response.Tatum Anderson (445\u2013446) reports on a feasibility trial in Liberia that provides hand-held ultrasound monitors to mothers in labour. Sabina Faiz Rashid talks to Andr\u00e9ia Azevedo Soares (447\u2013448) about anthropology, poverty, inequality and sex education in Bangladesh.Yanhong Gong et al. (449\u2013457) estimate over-the-counter and online pharmacy sales.Zheming Yuan et al. (484\u2013494) model the impact of the February 2020 lockdown of Wuhan City.Alessandra Ferrario et al. (467\u2013474) estimate sales volumes for chemotherapeutic agents.Helena Ojanper\u00e4 et al. (475\u2013483) study hand-hygiene compliance and nosocomial infections.Omotayo Fatokun (507\u2013508) explains how regulatory changes are designed to increase local production.Grace Li et al. (458\u2013466) find that most antibiotics are not sold in child-friendly formulations.Takahiko Koyama et al. (495\u2013504) analyse available sequence data. Jessica D Gipson et al. (505\u2013506) argue for the inclusion of infertility."} +{"text": "Kuramatsu et al. reviewed current therapies for reversal of new oral anticoagulants (NOACs) and anti-platelet agents in patients with acute intracerebral hemorrhage . In thei"} +{"text": "Correction to: BMC Cancer 21, 153 (2021)https://doi.org/10.1186/s12885-021-07842-4Following publication of the original article , the autThe correct author names are as follows:Athanasios (given name) Tampakis (family name)Ekaterini-Christina (given name) Tampaki (family name)Afrodite (given name) Nonni (family name)Ioannis D. (given name) Kostakis (family name)Alberto (given name) Posabella (family name)Konstantinos (given name) Kontzoglou (family name)Markus (given name) von Fl\u00fce (family name)Evangelos (given name) Felekouras (family name)Gregory (given name) Kouraklis (family name)Nikolaos (given name) Nikiteas (family name)The author group has been updated above and the original article has been"} +{"text": "In \u201cUnibody Endograft Using AFX 2 for Less Invasive and Faster Endovascular Aortic Repair: Protocol for a Multicenter Nonrandomized Study\u201d :e16959) there were two errors in the Collaborators List.The collaborator Sonia Ronchey was inadvertently not included in the collaborator list. Additionally, the collaborator name Pietro Volpet should have been listed as Pietro Volpe.The Collaborator List was initially published as follows:The LIVE Study Collaborators are as follows: Giancarlo Accarino; Dimitri Apostolou; Guido Bajardi; Stefano Bartoli; Filippo Benedetto; Franco Briolini; Stefano Camparini; Emidio Costantini; Giovanni Credi; Ruggiero Curci; Raffaello Dallatana; Gianmarco de Donato; Carlo Dionisi; Vittorio Dorrucci; Leonardo Ercolini; Gianfranco Fadda; Mauro Ferrari; Loris Flora; Andrea Gaggiano; Roberto Gattuso; Franco Grego; Sabrina Grimaldi; Giovanni Impedovo; Arnaldo Ippoliti; Antonio Jannello; Sergio Losa; Nicola Mangialardi; Isaac Martinez; Javier Martinez; Stefano Michelagnoli; Giancarlo Palasciano; Vincenzo Palazzo; Domenico Palombo; Raffaele Pulli; Giovanni Rossi; Antonino Scolaro; Gianantonio Simoni; Francesco Spinelli; Francesco Talarico; Maurizio Taurino; Marco Trogolo; Nicola Tusini; Gianfranco Veraldi; Pier Francesco Veroux; Gennaro Vigliotti; and Pietro Volpet.The Collaborator List has now been updated to the following:The LIVE Study Collaborators are as follows: Giancarlo Accarino; Dimitri Apostolou; Guido Bajardi; Stefano Bartoli; Filippo Benedetto; Franco Briolini; Stefano Camparini; Emidio Costantini; Giovanni Credi; Ruggiero Curci; Raffaello Dallatana; Gianmarco de Donato; Carlo Dionisi; Vittorio Dorrucci; Leonardo Ercolini; Gianfranco Fadda; Mauro Ferrari; Loris Flora; Andrea Gaggiano; Roberto Gattuso; Franco Grego; Sabrina Grimaldi; Giovanni Impedovo; Arnaldo Ippoliti; Antonio Jannello; Sergio Losa; Nicola Mangialardi; Isaac Martinez; Javier Martinez; Stefano Michelagnoli; Giancarlo Palasciano; Vincenzo Palazzo; Domenico Palombo; Raffaele Pulli; Sonia Ronchey; Giovanni Rossi; Antonino Scolaro; Gianantonio Simoni; Francesco Spinelli; Francesco Talarico; Maurizio Taurino; Marco Trogolo; Nicola Tusini; Gianfranco Veraldi; Pier Francesco Veroux; Gennaro Vigliotti; and Pietro Volpe.The correction will appear in the online version of the paper on the JMIR website on June 24, 2020, together with the publication of this correction notice. Because this was made after submission to PubMed, PubMed Central, and other full-text repositories, the corrected article has also been resubmitted to those repositories."} +{"text": "COVID-19 raakt de samenleving in zijn geheel. Het blijkt lastig om een goede balans te vinden tussen de risico\u2019s van een infectie met SARS-CoV\u20112 en de schade door de uitbraakbeheersing (verder: COVID-maatregelen). Meer bescherming voor de \u00e9\u00e9n betekent soms meer risico voor het leven, levensonderhoud en samenleven van een ander. Zicht op kwetsbare groepen is nodig om de zorg en ondersteuning te kunnen afstemmen op hun risico\u2019s en behoeften. Dit artikel gaat na welke groepen extra kwetsbaar zijn voor negatieve neveneffecten van COVID-maatregelen. We willen daarmee een discussie op gang brengen over optimale ondersteuning van deze en vergelijkbare groepen mensen tijdens en na deze crisis. COVID-maatregelen worden ge\u00efmplementeerd om mensen die risico lopen op ernstige COVID te beschermen . In maarCOVID-19 raakt allen, maar sommigen zijn om verschillende redenen extra kwetsbaar, zoals blijkt uit tab.\u00a0COVID-maatregelen beperken enerzijds ziekte en sterfte door SARS-CoV-2-infecties. Anderzijds leiden de maatregelen ook tot ziekte en sterfte. De gehandicaptenzorg, jeugdzorg, thuiszorg, wijkverpleging, geestelijke gezondheidszorg, huisartsenzorg, paramedische zorg en curatieve zorg zijn deels uitgesteld en deels vervangen door zorg op afstand. De omvang van de effecten op zorgbehoeftigen \u2013\u00a0die vrijwel per definitie verhoogd kwetsbaar zijn\u00a0\u2013 wordt gaandeweg duidelijk aan de hand van evaluaties , 5, 7, 9Er is sprake van langdurige stress, met mogelijk lichamelijke klachten, een toename van huiselijk geweld en extra middelengebruik , 5, 11. In de rapporten is vooral aandacht voor de persoonlijke ontwikkeling van jongeren en studenten, en het \u2018versomberen\u2019 van de samenleving. Sociaaleconomische gezondheidsverschillen en de kansenongelijkheid in de samenleving worden hierdoor versterkt.Het basisonderwijs is geruime tijd op afstand gegeven en kwam merendeels aan op ouders, met wisselende pedagogische vaardigheden en begeleidingstijd. Het opschorten van het (praktijk)onderwijs voor leerlingen in het voorgezet onderwijs, middelbaar beroepsonderwijs en hoger onderwijs heeft geleid tot studievertraging, verminderde motivatie en extra onzekerheid over werkgelegenheid , 6.Mogelijk nog belangrijker zijn de beperkingen voor de sociaal-emotionele ontwikkeling van jongeren, zowel binnen als buiten school. De effecten laten zich moeilijk voorspellen, maar de relatie van opleidingsniveau, gezondheidstoestand en levensverwachting is welbekend , 6. KwetVelen leven in angst en onzekerheid, en experts vrezen een toename van su\u00efcidepogingen . VerschiVeel COVID-19\u00a0pati\u00ebnten zijn in eenzaamheid gestorven en in de beleving van familie en vrienden ook zonder waardigheid. De beperkingen van het afscheid nemen en deelnemen aan een uitvaart kunnen het rouwproces negatief be\u00efnvloeden , 12.social distancing en thuiswerken beperken onze bewegingsvrijheid. Het dagelijks leven van wonen, werken, sport, cultuur en recreatie is aan banden gelegd. De openbare ruimte is een \u2018schaars goed\u2019 geworden. Sommigen hebben hier meer last van dan anderen. Denk aan stressoren als het begeleiden van schoolgaande kinderen, en het wegvallen van mantelzorgers en formele zorg [COVID-maatregelen als ele zorg , 3, 7. Dele zorg , 9.Dagbestedingsprogramma\u2019s voor specifieke doelgroepen zijn merendeels weggevallen. Dit raakt zowel cli\u00ebnten als hun begeleiders, voor een belangrijk deel vrijwilligers. Kwetsbare groepen zijn vooral kinderen/volwassenen met handicaps en/of psychische klachten, en ouderen , 6, 8, 9Sportactiviteiten, feestjes en bezoek zijn al geruime tijd niet meer vanzelfsprekend. Deze activiteiten bieden structuur en sociale contacten, dienen als uitlaatklep en kunnen bijdragen aan de zingeving van het leven . Er is nFysieke en sociale contacten zijn sterk ingeperkt. De financi\u00eble situatie, de bestaande (starters- en onderwater)problematiek op de woningmarkt en participatie in de samenleving zijn voor velen verslechterd . Er wordDe ongelijkheden in de samenleving worden groter. Zo loopt de betaalde arbeidsbijdrage van vrouwen terug ten opzichte van mannen en neemt het aantal werkloosheidsuitkeringen onder jongeren snel toe , 6, 9.De COVID-maatregelen beperken ook de mogelijkheden tot participatie in de samenleving. Asielzoekers en statushouders hebben nog meer moeite hun weg te vinden en zich een plek te verwerven in de Nederlandse samenleving . Dit gel\u2018Kwetsbare groepen\u2019 is een containerbegrip. Mensen zijn om verschillende redenen extra kwetsbaar voor COVID-maatregelen. Dit heeft consequenties voor de behoeften aan en mogelijkheden voor zorg en ondersteuning. Specificaties zijn nodig om het zorg- en ondersteuningsaanbod vorm te geven en te optimaliseren, waarbij er expliciete aandacht moet zijn voor risicocumulatie en toename van weerbarstige sociaaleconomische gezondheidsverschillen en kansenongelijkheid in bepaalde groepen .Elf adviesrapporten zijn nageplozen om zicht te krijgen op extra kwetsbare groepen voor COVID-maatregelen. Sommige kwetsbare groepen zijn breed omschreven, zoals ouderen, en omvatten meerdere subgroepen en neveneffecten, bijvoorbeeld verminderd zelfredzame ouderen die beperkingen ondervinden in het dagelijks leven en ouderen met een beperkt sociaal netwerk die lijden onder de inperking van sociale activiteiten. Andere groepen zijn juist smal omschreven en gerelateerd aan \u00e9\u00e9n neveneffect, zoals rouwverwerking door naasten van overledenen of participatie in de samenleving door ex-gedetineerden. Bij sommige groepen is al een stapeling van neveneffecten zichtbaar zie tab.\u00a0, bijvoorDe rapporten bieden een eerste uitsnede van de neveneffecten van en kwetsbare groepen voor COVID-maatregelen. Dit maakt een professionele discussie mogelijk over de concretisering van de zorg en ondersteuning aan kwetsbare groepen. Verdieping en verrijking is natuurlijk nodig. We hebben alleen Nederlandse adviesrapporten beoordeeld, die een nog geringe ervaringsbasis hebben en op veel aannamen zijn gebaseerd. Zo moet nog blijken in hoeverre jongeren een leer- en ontwikkelingsachterstand oplopen en in welke mate steunmaatregelen economische onzekerheden compenseren. De adviesrapporten vormen een momentopname en zullen snel gedateerd raken. Ook zullen nieuwe maatregelen gepaard gaan met nieuwe kwetsbare groepen. Zo dreef de anderhalvemetersamenleving blinden en slechtzienden in het nauw en versterkt de mondkapjescultuur inmiddels de isolatie van doven en slechthorenden. Sommige thema\u2019s in het maatschappelijke debat blijven onderbelicht in de adviesrapporten, zoals de coronakilo\u2019s (door het sluiten van de sportscholen), spirituele aspecten en milieueffecten. Ook ontbreekt aandacht voor bepaalde kwetsbare groepen, zoals gevangenen , illegalen en slachtoffers van mensenhandel (zorgdrempels), die al wel door Douglas et al. waren benoemd .De positieve neveneffecten van de COVID-maatregelen zijn in deze analyse buiten beschouwing gelaten. Zo hebben we geen aandacht besteed aan het feit dat driekwart van de mensen met psychoses juist minder problemen hebben ervaren , aan de Een checklist met effecten op het \u2018(samen)leven\u2019 is een goede eerste stap om de neveneffecten van COVID-maatregelen te categoriseren. De WHOQOL helpt om \u2018in de buurt te komen\u2019 van de doelstelling: snel zicht krijgen op extra kwetsbare groepen om tijdens en na een (infectieziekte)crisis de professionele discussie over zorg- en ondersteuning te voeden. De WHOQOL kent echter ook enkele beperkingen. Zo overlappen en be\u00efnvloeden de domeinen elkaar. \u2018Werk\u2019 valt bijvoorbeeld onder \u2018zelfstandigheid\u2019, maar \u2018financi\u00ebn\u2019 onder \u2018omgeving\u2019. Verder be\u00efnvloeden de lichamelijke, mentale en sociale gezondheidsaspecten elkaar onderling. Deze interacties worden niet direct inzichtelijk door screening van documenten via de WHOQOL. Het vraagt ook verdieping om zicht te krijgen op individuele variaties in kwetsbaarheden, coping-vaardigheden en veerkracht binnen groepen, wat essentieel is voor het organiseren van passende zorg en ondersteuning .De WHOQOL lijkt, ondanks de hier beschreven beperkingen, bruikbaar om een eerste systematisch overzicht te krijgen van de neveneffecten van de COVID-maatregelen. Analyse van elf rapporten maakt diverse bekende en nieuwe kwetsbare groepen zichtbaar. Pati\u00ebntverenigingen, belangenbehartigers en brancheverenigingen hebben verschillende signalementen en handreikingen uitgebracht waarmee de lijst kan worden verrijkt (groeimodel). Een overzicht van kwetsbare groepen biedt een basis om de monitoringsactiviteiten en informatievoorziening aan te scherpen, en waar nodig de contacten tussen (verondersteld) kwetsbare mensen, zorgverleners, GGD\u2019en, sociale wijkteams en overheden te versterken. Naarmate de groepen en hun kwetsbaarheden duidelijker in beeld komen, kan beter maatwerk worden geleverd. Denk aan het aanbieden van passende informatie over lichamelijke gezondheid en mentaal welbevinden, het organiseren van praktische hulp en het leveren van emotionele of financi\u00eble ondersteuning.De WHOQOL-vragenlijst kan, eventueel in aangepaste vorm, bijdragen aan verdiepend onderzoek binnen en tussen groepen die verhoogd kwetsbaar lijken voor de negatieve neveneffecten van COVID-maatregelen, en om de (langetermijn)effecten van COVID-19 te categoriseren. Wanneer we beter zicht hebben op kwetsbare groepen en specifieke kwetsbaarheden kunnen we in cocreatie inschatten welke mensen daadwerkelijk in de problemen komen en op welke manier zij het beste met passende ondersteuning kunnen worden bereikt. Dat vergt een netwerkinspanning van veel partijen: kwetsbare mensen zelf, zorgverleners en leerkrachten, andere professionals en overheden. Kwetsbare groepen dragen vrijwel per definitie de zwaarste lasten van rampen en crises. Door de handen ineen te slaan kunnen we de onevenredig zware last van COVID-maatregelen voor kwetsbare groepen enigszins met en voor hen beperken. Net zoals we dit proberen te doen voor groepen die verhoogd kwetsbaar zijn voor COVID-19."} +{"text": "Following publication of the original article (Huang et al. The original article (Huang et al."} +{"text": "Recently, Wandrer and colleagues published a study about the interaction of endoplasmic reticulum (ER) stress and autophagy and its relevance for hepatotoxicity (Wandrer et al. Drug-induced liver injury represents a major research field in toxicology (Godoy et al."} +{"text": "This month\u2019s special issue is on primary health care, introduced in an editorial by Tedros Adhanom Ghebreyesus (726). Shannon Barkley et al. (727) discuss what\u2019s needed to achieve the vision of primary health care and Teri A Reynolds et al. (728) describe the specifics of emergency, critical and operative care components.Gary Humphreys (731\u2013732) reports on the use of digital health services during the coronavirus pandemic and talks to Angela Nguku (733\u2013734) about the vital importance of community involvement in primary health care.Catherine Arsenault et al. (735\u2013746) examine the quality of hospital-based services.Tomas Zapata et al. (815\u2013817) detail lessons of effective strategies.Devaki Nambiar et al. (747\u2013753) field-test indicators.Luke N Allen et al. (754\u2013765) review the evidence on social determinants of noncommunicable diseases. Eliana E Kim et al. (766\u2013772) argue that essential surgery should be provided by family physicians.Emma Sacks et al. (773\u2013780) explain the link between community involvement and universal health coverage.Etienne V Langlois et al. (781\u2013791) count measures needed to strengthen care provision in low- and middle-income countries.Somtanuek Chotchoungchatchai et al. (792\u2013800) analyse contributions to the sustainable development goals.Kumanan Rasanathan & Tim G Evans (801\u2013808) recall the Declaration of Astana.Arit Udoh et al. (809\u2013811) detail a professional development plan for the pharmacy staff.Jan De Maeseneer et al. (812\u2013814) introduce a campaign for reassignment of 30% of health system funding.Sowmya Kadandale et al. (818\u2013820) uncover disconnects between primary care and climate policies."} +{"text": "Dear Editor,Scutellaria baicalensis Georgi. Wogonin contains various biological properties which include allergic diseases, anti-cancer therapy, and anti-inflammatory activities. Wogonin also shows the effects of removing toxins and cleansing the heart is a traditional naturally occurring flavonoid derived from the root extract of Chinese medicine, named al., 2019). Currenal., 2019; Ewendt al., 2019; Gao et al., 2019; Huang eal., 2019; Jiang eal., 2019; Jiao etal., 2019; Khan anal., 201915]; Khu; KhuScutal., 2019; Kim et al., 2019; Kong etal., 2019; Liang eal., 2019; Oomen eal., 2019; Tan et al., 2019; Wang anal., 2019; Wang etal., 201926]; Wan; WanScutal., 2019; Xing etal., 201929]; Zha; ZhaScutal., 2019).The authors declare no conflict of interest."} +{"text": "In the editorial section Chalapati Rao (298) emphasizes the need for accurate certification of cause-of-death. Bernardo Sousa-Pinto et al. (299) describe how the rates of testing for COVID-19 affect hospitalizations. In the news section, Tatum Anderson (302\u2013303) reports on the manufacturing processes needed to meet global demand once an effective vaccine for SARS-CoV-2 is developed. Amesh Adalja talks to Gary Humphreys (304\u2013305) about the lessons being learned in the COVID-19 epidemic.Stephanie Watson-Grant et al. (370\u2013372) identify an evaluation gap.Yanhong Jessika Hu & Benjamin John Cowling (360\u2013361) note reduction measures.Jason J Madan et al. (306\u2013314) provide an economic evaluation of short treatment regimens.Mohamed F Jalloh et al. (330\u2013340) study behavior change in an outbreak.Anne S Johansen et al. (353\u2013359) trace developments in primary health care.Charlotte James et al. (315\u2013329) estimate global incidence and prevalence Kalin Werner et al. (341\u2013352) review the evidence on cost-effectiveness of interventionsWilliam Mueller et al. (362\u2013364) propose standard epidemiological protocols. Alexandra Caulfield & Tobias Alfv\u00e9n (365\u2013366) argue for better prevention and treatment.Elizabeth Francis Beach et al. (367\u2013369) review measures to reduce risk of hearing damage."} +{"text": "De COVID-19-crisis en de intelligente lockdown hebben ertoe geleid dat het Nederlandse zorgsysteem maandenlang voor niet-COVID-pati\u00ebnten op slot kwam te zitten. Pati\u00ebnten durfden of konden niet naar hun huisarts, huisartsen waren terughoudend met doorverwijzingen naar het ziekenhuis en de zorg- en diagnostische processen werden vertraagd of aangepast. Dit gold ook voor kankerpati\u00ebnten. Hoe ernstig deze onderdiagnostiek is voor de prognose van de kankerpati\u00ebnt, hangt vooral af van kenmerken van de betreffende soorten kanker. In dit onderzoek hebben we de onderdiagnostiek in kaart gebracht met behulp van gegevens uit de Nederlandse Kankerregistratie en het Pathologisch-Anatomisch Landelijk Geautomatiseerd Archief. Vanaf de week waarin de eerste COVID-19-pati\u00ebnt in Nederland werd gediagnosticeerd, is een daling van 20\u201340% in het aantal kankerdiagnoses te zien. De daling geldt voor vrijwel alle tumorsoorten, ook voor kankers waarbij het niet tijdig stellen van de diagnose levensbedreigend is, zoals bij pati\u00ebnten met kanker in de luchtwegen (gemiddeld 23%) en het hoofd-halsgebied (gemiddeld 36%), en bij hematologie (gemiddeld 26%). Er moet meer aandacht komen voor de gesignaleerde onderdiagnostiek en de rol van de eerstelijnszorgverleners, zoals huisartsen en tandartsen. Daarbij is het zeer belangrijk dat pati\u00ebnten altijd het vertrouwen houden dat ze bij klachten hun zorgverlener kunnen raadplegen. De COVID-19-crisis en de intelligente lockdown, die verdere verspreiding in Nederland moet voorkomen, hebben ertoe geleid dat het Nederlandse zorgsysteem maandenlang voor niet-COVID-pati\u00ebnten op slot kwam te zitten. De prioriteit in de zorg kwam te liggen op het cre\u00ebren van voldoende capaciteit van de intensive care (IC), de zwakste schakel in het zorgsysteem tijdens de COVID-19-pandemie.delay kan worden veroorzaakt door de (potenti\u00eble) pati\u00ebnt, de zorgverlener en/of het zorgsysteem. Het stellen van vragen over de redenen van uitstel is voor de toekomst extreem belangrijk, om zo uitstel te voorkomen. In het geval van COVID-19 is er een aantal factoren aan te wijzen op basis van data uit de Nederlandse Kankerregistratie (NKR) en het Pathologisch-Anatomisch Landelijk Geautomatiseerd Archief (PALGA) zien dat het aantal kankerdiagnoses aanzienlijk was gedaald , 2. De oTijdens de eerste intelligente lockdown daalde het aantal huisartsconsulten met 70% en het aantal verwijzingen naar het ziekenhuis met 75%, en halveerde het aantal nieuwe kankerdiagnoses \u20135.Hoe ernstig deze onderdiagnostiek is voor de prognose van de kankerpati\u00ebnt, hangt vooral af van kenmerken van de betreffende kankers. Zo maakt het uit in welk stadium de kanker zich bevindt, wat de groeisnelheid en agressiviteit van de kanker is en wat de behandelmogelijkheden zijn voor de betreffende pati\u00ebnt. Voor kanker met een relatief gunstige prognose, zoals borst- en huidkanker, is een tijdelijk delay mogelijk minder \u2018erg\u2019 dan voor ziekten waarvan snelle, tijdige diagnostiek van levensbelang kan zijn. We denken hierbij aan aandoeningen als long-, keelholte-, slokdarm-, alvleesklier-, eierstok- en maagkanker, en een aantal hematologische kankers .Het is belangrijk om eerst inzicht te krijgen in de mate van onderdiagnostiek in zijn totaal, om vervolgens de mate van onderdiagnostiek voor diverse subgroepen in kaart te brengen. Inzichten in onderdiagnostiek voor de diverse tumorsoorten en regio\u2019s kunnen bijvoorbeeld voorspellen waar er later een piek in het aantal diagnoses te verwachten is. Daarnaast kunnen we mogelijk ook lessen trekken voor de toekomst op basis van de genoemde factoren en andere factoren, zoals leeftijd.We hebben pati\u00ebnten ge\u00efncludeerd uit de Nederlandse Kankerregistratie (NKR) van wie op basis van diagnoses uit PALGA bekend is dat ze in de eerste 23\u00a0weken van 2020 (tot en met 6\u00a0juni) gediagnosticeerd zijn met een invasieve vorm van kanker (data-extractie vond plaats op zondag 21\u00a0juni 2020). Voor vergelijking is gekeken naar de diagnoses uit dezelfde periode in 2019 om de mogelijke invloed van andere factoren, zoals jaargetijde en vakantieperiodes zichtbaar te maken. Er is een identieke data-extractie uit de NKR gemaakt op zondag 23\u00a0juni 2019. Dit was noodzakelijk omdat de diagnoses die momenteel van 2020 bekend zijn, grotendeels bestaan uit voorlopige registraties en in de loop van de tijd nog kunnen veranderen nadat aanvullend onderzoek is verricht. Dit laatste heeft vooral invloed op de stadi\u00ebring van de kanker.De analyses zijn beschrijvend van aard. We hebben gekeken naar de relatieve verandering in kankerdiagnoses tijdens de COVID-19-periode. Hiervoor is het aantal kankerdiagnoses per week vergeleken met het gemiddelde aantal kankerdiagnoses in week 2\u00a0tot en met 5\u00a0van 2020 en uitgedrukt als percentage van dit gemiddelde. Naast het totale aantal diagnoses is er een uitsplitsing gemaakt naar type kanker, leeftijdscategorie en provincie.Voor de gestratificeerde analyses is het totale aantal diagnoses van meerdere weken genomen , omdat de aantallen per week te weinig inzicht kunnen geven in de trends van diverse subgroepen. De aantallen zijn hier ook weer vergeleken met het aantal diagnoses in week 2\u00a0tot en met 5. Voor de uitsplitsing naar tumorsoort is gekozen voor een indeling op algemene hoofdgroep, met uitzondering van kanker in de mondholte/orofarynx en hematologische maligniteiten. Voor andere kankersoorten werden de aantallen per week bij verdere opsplitsing te klein om trends te kunnen weerspiegelen.In fig.\u00a0De daling is voor vrijwel alle tumorsoorten te zien fig.\u00a0, met de In fig.\u00a0De gesignaleerde daling geldt voor alle leeftijdscategorie\u00ebn fig.\u00a0, maar isDe daling is in alle provincies te zien fig.\u00a0, inclusiDit onderzoek beschrijft de relatief hoge mate van onderdiagnostiek in Nederland tijdens de eerste paar maanden van de COVID-19-pandemie. Dit is te zien voor alle kankersoorten, alle leeftijden en in alle provincies. Voor enkele groepen, zoals kankers van de luchtwegen, het hoofd-halsgebied en hematologie, is het missen van diagnoses van direct levensbelang.Deze daling zal grotendeels toe te schrijven zijn aan de COVID-19-pandemie, maar er zijn wel week tot week fluctuaties zichtbaar. In de eerste weken van het jaar (onze referentieperiode) liggen de aantallen doorgaans wat hoger vanwege een inhaalslag na de kerstvakantie. Ook spelen feestdagen een rol in de fluctuaties. Voor kleinere tumorsoorten is het ook lastiger om conclusies te trekken. Daarom hebben we de trends met 2019 vergeleken en meerdere weken samengenomen.De bevolkingsonderzoeken voor borst-, baarmoederhals- en darmkanker zijn stopgezet in week\u00a012, maar er zit een aantal weken tussen het bevolkingsonderzoek en de pathologische bevestiging van een tumor [In week 15\u00a0hebben diverse partijen, waaronder IKNL, gezamenlijk een oproep gedaan om zich met klachten eerder bij de huisarts te melden. Dit heeft bij geen van de onderzochte groepen een duidelijk effect gehad , 9. Een Diverse factoren kunnen mogelijk bijdragen aan het voorkomen van het wegblijven van mogelijke pati\u00ebnten bij de eerstelijnszorg. Ten eerste is het van belang dat pati\u00ebnten voldoende kennis van mogelijke klachten en symptomen hebben en dit benadrukken als ze hulp zoeken, bijvoorbeeld door de huisarts te bellen. Klachten vari\u00ebren uiteraard per tumorsoort en kunnen ook bij andere ziekten voorkomen. Voor diverse tumorsoorten is het ook lastig om de diagnose te stellen, omdat er (nog) geen klachten of symptomen zijn. Bij andere soorten zijn er klachten die om alertheid vragen. De volgende klachten kunnen wel duiden op een maligniteit: bloed in slijm, ontlasting of urine, (moeder)vlekken die nieuw zijn of veranderen, een verdikking of knobbeltje op de huid of in het lichaam, gewichtsverlies of moeheid zonder reden . KWF KanOp het moment dat we dit schrijven vindt er opnieuw afschaling van de reguliere zorg plaats vanwege een tweede COVID-19-piek. Op- en afschaling van de zorg gebeurt aan de hand van een lijst met gestelde diagnoses waarbij tijdige diagnostiek dus niet als criterium is opgenomen , 9. De lOok in andere landen heeft de COVID-19-pandemie geleid tot een delay in kankerdiagnoses. In de Verenigde Staten is het aantal kankerdiagnoses tot eind april 2020 met 65% verminderd , 12. EenIn welke mate de extra delay door de COVID-19-pandemie impact heeft op de uitkomsten voor de pati\u00ebnt is nog niet duidelijk. Dit zal ook afhangen van andere factoren, waaronder het type kanker, het stadium waarin de kanker zich bevindt, de groeisnelheid en agressiviteit van de tumor, hoe goed het type kanker te behandelen is, maar ook of en hoeveel later de pati\u00ebnt uiteindelijk de diagnose heeft gekregen. Dergelijke vragen, en de vraag welke impact de COVID-19-pandemie op de behandeling van kankerpati\u00ebnten heeft gehad, zullen in vervolgonderzoeken worden beantwoord.In het Verenigd Koninkrijk zijn reeds modelmatige berekeningen gedaan naar de impact van de COVID-19-epidemie op de delay in kankerdiagnoses en de veranderingen in de zorgverlening, berekend vanaf het punt van de diagnosestelling. In deze berekeningen zijn ook het aanpassen, uitstellen of weglaten van behandelingen meegenomen . De auteNu we in de tweede COVID-19-golf zitten, zou er meer aandacht aan de diagnostiek van andere ziekten, zoals kanker, besteed moeten worden. In de zomer was het aantal kankerdiagnoses ongeveer gelijk aan het aantal van 2019 en in september is er een inhaalslag geweest . Op dit In dit onderzoek laten we zien dat het aantal kankerdiagnoses tijdens de eerste lockdown is achtergebleven bij wat we verwachtten. Wat de exacte gevolgen hiervan zijn moet nader onderzoek uitwijzen. Het is belangrijk dat de drempel om eerstelijnszorg te zoeken voor pati\u00ebnten wordt weggenomen en dat de reguliere zorg zo goed mogelijk doorgang moet vinden."} +{"text": "In the editorial section, Vasee Moorthy et al. (150) announce a WHO pre-print server for research on the novel coronavirus (2019-nCoV). Peter Beyer & Sarah Paulin (151) provide an update on priority pathogens and the antibiotic pipeline. Payao Phonsuk et al. (152) announce an upcoming theme issue and call for papers on the health impacts of climate change and geopolitics. Darren B. Taichman et al. (153) propose a new disclosure form for the authors submitting work to medical journals.In the news section, Lynne Eaton (156\u2013157) reports on global supply chain problems with heparin and magnesium sulfate. Karla Soares-Weiser (158\u2013159) talks to Gary Humphreys about systematic reviews of evidence and the Cochrane Library.Rebecca Knowles et al. (176\u2013188) collate data from 20 low- and middle-income countries.Andres I Vecino-Ortiz & Deivis N Guzman-Tordecilla (169\u2013175) study gun-carrying restrictions and gun-related mortalityShatrunajay Shukla et al. (207\u2013212) describe the expansion of regulatory scope to medical devices. Benjamin TB Chan et al. (160\u2013168) evaluate measures to improve quality of care for patients with chronic diseases. Worarat Imsanguan et al. (83\u201384) provide infection prevention recommendations for contacts of people with tuberculosis.Hugo Perazzo et al. (189\u2013198) review the evidence of effectiveness for generic direct-acting agents.Taenia solium control strategies.Matthew A Dixon et al. (199\u2013206) model the effects of Gautam Bhan (220\u2013222) examine the links between informal work and maternal and child health.David H Jernigan & Pamela J Trangenstein (223\u2013225) explore next steps for WHO\u2019s global alcohol strategy.James L Nuzzo & James Steele (226\u2013227) argue for a causal systems map.Harry Rutter et al. (227\u2013228) outline systems approaches to support physical activity."} +{"text": "Dear Editor,Apigenin belongs to the group of flavonoids positioned on the backbone of 2-phenylchromen-4-one (2-phenyl-1-benzopyran-4-one) and is most extensively allocated in herbs, vegetables, and fruits . Here, al., 2017; Britto al., 2017; Charalaal., 2017; Chen etal., 2017, 20206]in vivo aal., 2017; Dean etal., 2017; Feng etal., 2017; Ganai, al., 2017; Han et al., 2017; Hassan al., 2017; He et aal., 2017; Jiang eal., 2017; Jing etal., 2017; Kang etal., 2017; Ketkaewal., 2017; Lee et al., 2017; Li et aal., 2017, 2019[29al., 2017; Liu et al., 2017; Lu et aal., 2017; Malik eal., 2017; Mirzoeval., 2017; Mrazek al., 2017; Nelson al., 2017; Pang etal., 2017; Qiu et al., 2017; Quan etal., 2017; Ra\u0161kovial., 2017; Ren et al., 2017; Safari al., 2017; S\u00e1nchezal., 2017; Sang etal., 2017; Sharma al., 2017; Siddiqual., 2017; Stump eal., 2017; Thangaial., 2017; Tong etal., 2017; Wang etal., 2017, 2018[56al., 2017; Wu et aal., 2017; Xu et aal., 2017; Zare etal., 2017; Zhang eal., 2017, 2018[63al., 2017, 2020[60al., 2017; Zhong eal., 2017, 2018[66al., 2017).This research was supported by Golden Seed Project (213006051WTE11) funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA), the Ministry of Oceans and Fisheries (MOF), the Rural Development Administration (RDA), and the Korea Forest Service (KFS), Republic of Korea.The authors declare no conflict of interest."} +{"text": "In the editorial section, Viroj Tangcharoensathien et al. (78) present research on COVID-19 response and mitigation measures, the focus of this month\u2019s issue and of the 2021 Prince Mahidol Award Conference.In the news section, Gary Humphreys and Sophie Cousins report (81\u201382) on initiatives to reduce the environmental impact of the health sector. Poornima Prabhakaran talks to Andr\u00e9ia Azevedo Soares (83\u201384) about climate-related health hazards in India, initiatives to address them, and the challenges presented by industrial development. Huajie Jin et al. (112\u2013124) estimate cost-of-illness.Vanessa W Lim et al. (92\u2013101) report findings from cohort surveys.Constance RS Mackworth-Young et al. (85\u201391) analyse community perspectives.Niheer Dasandi et al. (102\u2013111) examine intergovermental engagement on health impacts of climate change.Yevgen Nazarenko et al. (125\u2013137) compare air quality standards.Johannes K\u00f6hler et al. (138\u2013147) study the practices of national ethics and bioethics committees.Jennifer Cole and Klaus Dodds (148\u2013154) ask if the COVID-19 response can reform climate change.Teck Chuan Voo et al. (155\u2013161) review the ethical implications of immunity certificates in the context of COVID-19.Ann Dulhanty (162\u2013163) discusses ethics of data collection and use.Freddy Sitas et al. (164\u2013165) point to a need for better data and public information about the effects of smoking on outcomes of respiratory infections.Albert Persaud et al. (166\u2013168) describe geopolitical factors affecting health."} +{"text": "In editorials this month, Ren Minghui et al. (582) note gaps in access to medicines for noncommunicable diseases and mental health conditions. Amiya Bhatia et al. (583) call for prevention of violence against children associated with school closures and movement restrictions.Tatum Anderson (586\u2013587) reports on shortages of medical oxygen needed to treat people hospitalized with severe COVID-19. Melanie Saville talks to Gary Humphreys (588\u2013589) about the challenges in developing and distributing future vaccines for SARS-CoV-2 vaccines.Cornelia A Barth et al. (599\u2013614) characterize users of rehabilitation services in post-conflict settings. Haijiang Lin et al. (632\u2013637) detail COVID-19 control strategies in Taizhou city. Dirk Engels et al. (615\u2013624) propose an integrated research agenda. Andreas Deckert et al. (590\u2013598) compare strategies for pooled-sample analysis.Jerome Amir Singh et al. (625\u2013631) detail effects on participants. Rebecca Grant et al. (638\u2013640) discuss models showing epidemic potential for monkeypox. Hannah Kettler et al. (641\u2013643) enumerate interventions needed to improve access."} +{"text": "In the editorial section, James Campbell and Fahrettin Koca (2) discuss the support needed for the health and care workforce highlighted by the COVID-19 pandemic. Jozef Bartovic et al. (3) point to the need for equitable access to vaccines for refugees and migrants.In the news section, Lynn Eaton (6\u20137) reports on how pandemic-response measures, including school closures and movement restrictions, are impacting the sexual and reproductive health and rights of adolescent girls. Fadi Meroueh talks to Andr\u00e9ia Azevedo Soares (8\u20139) about ensuring health equity and harm reduction services in prisons.Tedbabe Degefie Hailegebriel et al. (69\u201371) follow implementation of kangaroo mother care.Zheng Hui et al. (10\u201318) model infection control measures.Aisha Nandini Zoe Dasgupta (67\u201368) analyse terms used to refer to contraception.Kyung-Bok Son et al. (62\u201366) report the severity classification used to group patients.Andrew T Boyd et al. (34\u201340) study measures to prevent HIV/TB coinfection.Son N Do et al. (50\u201361) study outcomes in a prospective cohort.John PA Ioannidis (19\u201333) estimates the infection fatality rate of COVID-19.Anne Marie Thow et al. (41\u201349) discuss implications for farmers."} +{"text": "In the editorial section, Qais Alemi et al. (510) discuss what\u2019s needed to reduce pandemic risks for refugee populations. Heather Adair-Rohani (511) portrays the work of Kirk Robert Smith in improving air quality. In the news section, Gary Humphreys (514\u2013515) reports on how regulatory agencies are adapting to the need for expedited development and distribution vaccines, tests and treatments for COVID-19. Baruch Fischhoff talks to Fiona Fleck (516\u2013517) about behavioural science and explains why governments need to test the messages used in the context of pandemic response. Julien Cappelle et al. (539\u2013547) document Nipah virus circulation.Rosanna Lyus et al. (530\u2013538) evaluate national registration of essential antimicrobials.Steffie Heemelaar et al. (548\u2013557) report maternal near-miss surveillance data.P. falciparum.Rebecca Thomson et al. (558\u2013568) review the evidence for genetic changes in Shoi Shi et al. (518\u2013529) estimate the impact of travel restrictions.Anila Jacob et al. (576\u2013578) argue for inclusion of the ecosystem.Mellick J Chehade et al. (569\u2013575) propose personal health hubs.Karin R Jongsma and Fleur Jongepier (579\u2013580) explore value-sensitive design."} +{"text": "In editorials on antimicrobial resistance, Peter Beyer et al. (822) announce a new research and development fund and Timo Minssen et al. (823) examine social, cultural and economic aspects of antimicrobial overuse.In the news section, Andr\u00e9ia Azevedo Soares (826\u2013827) describes efforts to ensure equitable access to future vaccines for SARS-CoV-2. Cheryl Cohen talks to Gary Humphreys (828\u2013829) about the response to the COVID-19 pandemic and surveillance for respiratory infections in South Africa.Juan Li et al. (830\u2013841) detail the function of Fangcang shelter hospitals in Wuhan.Yibeltal Assefa et al. (894\u2013905) review evidence of primary health care impacts.Adam Gyedu et al. (869\u2013877) study availability of health insurance and surgery.Mhinjine Kim et al. (842\u2013848) describe the protocols used to make a dedicated COVID-19 hospital.Muchiri E Wandai (878\u2013885) establish trends in normal, overweight and obese categories. Anna D Gage et al. (849\u2013858) assess quality improvement measures in health facilities.Emmanuela E Ambrose (859\u2013868) re-use dried blood spots to measure prevalence of sickle cell trait and disease. Azeb Tesema et al. (906\u2013908) list changes needed to cover noncommunicable diseases.Christian Kraef et al. (886\u2013893) focus on the cause of 1 in 5 global deaths."} +{"text": "The editorial team is grateful to all the experts who dedicated their time to peer-review our articles over the past 12 months. We wish to thank them and all those whose names are not listed below but gave helpful advice and support. We apologise to any reviewers whose names we may have missed.Our reviewers in 2020 were:Mohamed Abbas; Stephan Aberle; Muna Abu Sin; Mary Adams; Karam Adel Ali; Cornelia Adlhoch; Karoline Aebi-Popp; Seven Johannes Sam Aghdassi; Antonella Agodi; Lis Alban; Jan Albert; Sheikh Taslim Ali; Franz Allerberger; Erik Alm; Christian Althaus; Matthias an der Heiden; Maria an der Heiden; Claire Anderson; Brito Anderson; Raiees Andrabi; Nick Andrews; Xanthi Andrianou; Denise Antona; Alberto Antonelli; Habibollah Arefian; Wouter Arrazola de O\u00f1ate; Tommi Asikainen; Olle Aspevall; Barry Atkinson; Erik Aurell; Sabrina Bacci; Agoritsa Baka; Craig Baker-Austin; Tamas Bakonyi; Sooria Balasegaram; Koye Balogun; Miguel Bao-Dominguez; Osamah Barasheed; Cyril Barbezange; Ian Barr; Luisa Barzon; Denis Bauer; Gabrielle Beaudry; Julien Beaut\u00e9; Cecile Beck; Philippe Belanger; Edward Belongia; Justus Benzler; Anne Berger-Carbonne; Isha Berry; Marianne Besnard; Philippe Beutels; Stuart Blacksell; Hubert Blain; Evan M. Bloch; Seth Blumberg; Nicki Boddington; Pierre-Yves Boelle; Magnus Boman; Idesbald Boone; Martin Bootsma; David Boulware; Nicola Luigi Bragazzi; Arne Brants\u00e6ter; Eeva Broberg; Stefan Brockmann; Tal Brosh-Nissimov; Lisa Brouwers; Kevin Brown; Mathias Bruyand; Robin Bruyndonckx; Mia Brytting; Udo Buchholz; Niccol\u00f2 Buetti; Nicholas Bundle; Thierry Burnouf; Karen Burns; Valeria Cagno; Worth Calfee; Alessandra Carattoli; Yehuda Carmeli; AnnaSara Carnahan; Gail Carson; Jordi Casabona; Geraldine Casey; Jesus Castilla Catalan; Giancarlo Ceccarelli; Andr\u00e9 Charlett; Remi Charrel; Yee-Chun Chen; Hua Chen; Nikoloz Chkhartishvili; Eun Hwa Choi; Anissa Chouikha; Gerardo Chowell; Daniel Chu; Bruno Ciancio; Sandra Ciesek; Milan Cizman; Eric C. J. Claas; Hannah Clapham; Luigi Codecasa; Rosalind Lucy Coleman; Vittoria Colizza; Alex Cook; Teresa Coque; Samuel Cordey; Victor Corman; Laura Cornelissen; Benjamin Cowling; Natasha Crowcroft; Fortunato D'Ancona; Gavin Dabrera; Niklas Danielsson; Fran\u00e7ois Danion; Costas Danis; Masoud Dara; Siddhartha Sankar Datta; Helena de Carvalho Gomes; Birgitta de Jong; Ana Maria de Roda Husman; Gaston De Serres; Gerard de Vries; Genevieve Deceuninck; Valerie Delpech; David Denning; Sara Dequeker; Jean-Claude Desenclos; Simona Di Giambenedetto; Sabine Diedrich; Michaela Diercke; Joakim Dillner; W Dimech; Gerhard Dobler; Roger Dodd; Jennifer Dowd; Timothee Dub; Sandra Dudareva-Vizule; Dominic Dwyer; Isabella Eckerle; Tim Eckmanns; Michael Edelstein; Paul Effler; Maren Eggers; Martin Eichner; Jorge Eiras; David Elliman; Mallory Ellingson; Andrea Endimiani; Theresa Enkirch; Daniel Faensen; Darryl Falzarano; Ugo Fedeli; John Ferguson; Virginie Ferre; Siri Laura Feruglio; Julie Figoni; Antonietta Filia; Bastian Fischer; David Fisman; Brendan Flannery; Per Follin; Anders Fomsgaard; Tony Fooks; Ivo Foppa; Frode Forland; Anne Fouillet; Sandra Fournier; Greg Fox; Elisabetta Franco; Alexander Friedrich; Ingrid Friesema; Shouichi Fujimoto; Wolfgang Gaissmaier; Juan Carlos Galan; Gary Garber; Michel Garenne; Petra Gastmeier; Raphael Gaudin; Marius Gilbert; Rosina Girones; John Glasser; Shana Godfred-Cato; Pere Godoy; Emily Goldstein; Maureen Goss; C\u00e9line Gossner; Magnus Gottfredsson; Luigi Gradoni; Gilbert Greub; Margrethe Greve-Isdahl; Larisa Gubareva; Jan Gunst; Jean-Paul Guthmann; Bernardo Rafael Guzman Herrador; Katrine Bach Habersaat; Christos Hadjichristodoulou; Anette Hammerum; Sonja Hansen; Pia Hardelid; Sally Hargreaves; Heli Harvala; Joana Haussig; Craig Hedberg; Janneke Heijne; Joris Hemelaar; Rafael Herruzo; Markus Hilty; Hans Hirsch; Boris Hogema; Heidemarie Holzmann; Susan Hopkins; Rachel Hoyles; Wan-Ting Huang; Jim F Huggett; Anette Hulth; Hilary Humphreys; Daniel Hungerford; Michael H\u00f6hle; Judith H\u00fcbschen; Thomas Inns; Giuseppe Ippolito; Nazrul Islam; Esma Islamaj; Angela Iuliano; Alexandra Jablonka; Dorota Jamrozy; Claire Jenkins; Eric A Jensen; Silvia Jimenez-Jorge; Kari Johansen; Steven John; Helen Johnson; Terry Jones; Pernille Jorgensen; Laurence Josset; Charlotte S. J\u00f8rgensen; Sarah Kada; Kamran Kadkhoda; Alexander Kallen; G\u00fcnter Kampf; Anu Kantele; Tommi Karki; Alyson Kelvin; Maria Keramarou; Liliane Keros; Marie E. Killerby; Pete Kinross; Esther Kissling; Don Klinkenberg; Gwen Knight; Anke Kohlenberg; Marion Koopmans; Uwe Koppe; Rolf Kramer; Florian Krammer; Tyra Krause; Gerard Krause; Annelies Kroneman; Adam Kucharski; Tanja Kuchenmuller; Peter Kuhnert; Ed J Kuijper; Michael Kurilla; Kin On Kwok; Kathryn E. Lafond; Filippo Lagi; Amparo Larrauri; Eric Lau; Jean-Philippe Lavigne; Jeffrey Lazarus; Quentin Le Hingrat; Jason J Leblanc; Jonggul Lee; Clara Lehmann; Sebastian Lequime; Tinne Lernout; Kathy Leung; Daniel Levy-Br\u00fchl; Bruno Lina; Theodore Liou; Marc Lipsitch; Yang Liu; Giovanni Lo Iacono; Anna-Leena Lohiniva; SW Long; Rohit Loomba; Nicola Low; Christian Lueck; Barbara M Lund; \u00c5ke Lundkvist; Xufei Luo; Theodore Lytras; Outi Lyytik\u00e4inen; Emma L\u00f6f; Renke L\u00fchken; Pablo M De Salazar; Kristine Macartney; Ian Mackay; Gkikas Magiorkinis; Cecile Magis-Escurra; Benjamin Maier; Gannon C.K. Mak; Helena Maltezou; Michal Mandelboim; Annette Mankertz; Jean-Michel Mansuy; Josefa Masa-Calles; Mathieu Mateo; Sebastian Maurer-Stroh; Noel McCarthy; Gerald McInerney; Genevieve McKew; Lucy McNamara; Jolyon Medlock; Adam Meijer; Paula Meireles; Jacques Meis; Kassiani Mellou; Jeffrey Mercante; Lore Merdrignac; Mathias Merker; Stefano Merler; Silvia Meschi; Benjamin Meyer; Kai Michaelis; Jean Millet; Vivi Miriagou; Kenji Mizumoto; Chris Ka Pun Mok; Israel Molina; Jacob Moran-Gilad; Yamir Moreno; Clint Morgan; Keita Morikane; Shila Mortensen; Jo\u00ebl Mossong; Sandra Mounier-Jack; Vincent Munster; David Murdoch; K. Darwin Murrell; Michelle Murti; Mette Myrmel; Olaf M\u00fcller; Celine Nadon; Anna Nagy; Ndeindo Ndeikoundam Ngangro; Howard Needham; Joice Neves Reis Pedreira; Nathalie Nicolay; Jens Nielsen; Jessica Nihl\u00e9n Fahlquist; Hiroshi Nishiura; Harold Noel; Paulo Nogueira; Hanna Nohynek; Torbj\u00f6rn Nor\u00e9n; Angela Novais; Glen Nowak; Norbert Nowotny; Dennis Nurjadi; Dominik Sebastian N\u00f6rz; Anna Odone; Lulla Opatowski; W Paget; Jos\u00e9 Arthur Paiva; Takis Panagiotopoulos; Marcus Panning; Annalisa Pantosti; Anna Papa; John Papp; Gabriel Parra; Suzan Pas; Marie-Claire Paty; Lara Payne; Richard Pebody; Ana Penedos; Pasi Penttinen; Ranawaka A P M Perera; Ana Perez-Escoda; Unai Perez-Sautu; Martin Petric; Susanne Pfefferle; Yvonne Pfeifer; Anastasia Pharris; Gorben Pijlman; Johann Pitout; Diamantis Plachouras; Pedro Plans-Rubi\u00f3; Evangelia D. Platsouka; Mateusz Plucinski; Catherine Pl\u00fcss-Suard; Laurent Poirel; Chiara Poletto; Constantina Politis; Mario Poljak; Marina Pollan; Stefano Pongolini; Leo Poon; MahmoudReza Pourkarim; Andrew Preston; Natalie Prystajecky; Andrea Pugliese; Milo Puhan; Joan Puig-Barbera; Billy Quilty; Filip Raciborski; Barbara Rath; Andrainolo Ravalihasy; Raffaella Ravinetto; Vincenza Regine; Chantal Reusken; Ute Rexroth; Giovanni Rezza; Matteo Ricc\u00f2; Daniel Richardson; Michael Rigby; Thomas V. Riley; Emmanuel Robesyn; Joacim Rockl\u00f6v; Angela Rose; Maiken Rosenstierne; Paul Rota; Adam Roth; Maja Rupnik; Katilyn Sadtler; Marc Saez; Linda Saif; Cristiano Salata; Gloria Sanchez; Sabine Santibanez; Gianpaolo Scalia-Tomba; Samuel Scarpino; Barbara Schimmer; Jonas Schmidt-Chanasit; Stefan Scholz; Madlen Schranz; Peter Schulz; Mitchell Schwaber; Sheree Schwartz; Ilan Schwartz; Ettore Severi; Anita Shah; Giri Shankar; Yehuda Shoenfeld; Aaron Siegler; Carlo Signorelli; Lotta Siira; Ian Simms; Andreas Sing; Anika Singanayagam; Mary Anissa Sinnathamby; Hans-Christian Slotved; Patrick Soentjens; Kristian Soltesz; Gianfranco Spiteri; Anneke Steens; Paola Stefanelli; James Stimson; Alexander J Stockdale; Heribert Stoiber; Maja Stosic; Julia Stowe; Sunita Sturup-Toft; Kanta Subbarao; Bertrand Sudre; Carl Suetens; Aya Sugiyama; Jonathan Suk; Sheena Sullivan; Patrick Sullivan; Arnfinn Sundsfjord; Piyarat Suntarattiwong; Makoto Takeda; Johanna Takkinen; Lara Tavoschi; Karin Tegmark-Wisell; Anders Tegnell; Peter Teunis; Robin Thompson; Craig Thompson; Andrew Thompson; Stacy Todd; Laura Toivonen; Kristin Tolksdorf; Sarah Tonkin-Crine; Karina Top; Nuria Torner; Arturo Torres Ortiz; Alma Tostmann; Ramona Trebbien; Tim Tsang; Sarah Tschudin-Sutter; Gro Tunheim; Soren Uldum; Helmut Uphoff; Palle Valentiner-Branth; Sophie Valkenburg; Birgit van Benthem; Gerrita van den Bunt; Kees. C. van den Wijngaard; Wim van der Hoek; Nicoline van der Maas; Kristien Van Reeth; Edward van Straten; Guido Vanham; Philippe Vanhems; A Vantarakis; Olli Vapalahti; Alkiviadis Vatopoulos; Aurelie Velay; Lasse Vestergaard; C\u00e9cile Viboud; Peter Vickerman; Russell M Viner; Leo Visser; Margreet Vos; Georgia Vourli; Bea Vuylsteke; Silja V\u00f6neky; Benjamin Wachtler; Jacco Wallinga; Jan Walter; Gilles Wandeler; John Watkins; Scott Weese; Matthijs Welkers; Tobias Welte; Guido Werner; Therese Westrell; Andreas Widmer; Joanne Wildenbeest; Annelies Wilder-Smith; Emma Wiltshire; Christian Winter; Carl-Heinz Wirsing von K\u00f6nig; Charles Wiysonge; Katja Wolthers; Anabelle Wong; Joseph Wu; Jianhong Wu; Roman W\u00f6lfel; Andrea W\u00fcrz; Guogang Xu; Takuya Yamagishi; Jonathan Yoder; Dong Keon Yon; Barnaby Edward Young; Ioannis Zabetakis; Maria Zambon; Lorenzo Zammarchi; Philipp Zanger; Robert Zangerle; Peter Zarb; David Zella; Werner Zenz; Xu-Sheng Zhang; Walter Zingg; Phillip Zucs."} +{"text": "Correction to: BMC Infect Dis 18, 677 (2018)https://doi.org/10.1186/s12879-018-3565-zFollowing publication of the original article , the autBelow the GLIMP Study Group authors\u2019 names are given:Argentina\u2014Patricia Karina Aruj ; Silvia Attorri ; Enrique Barimboim ; Juan Pablo Caeiro, Mar\u00eda I Garz\u00f3n ; Victor Hugo Cambursano ; Adrian Ceccato ; Julio Chertcoff, Florencia Lascar, Fernando Di Tulio ; Ariel Cordon D\u00edaz ; Lautaro de Vedia ; Maria Cristina Ganaha ; Sandra Lambert ; Gustavo Lopardo, Hospital Bernardo Houssay, Vicente L\u00f3pez, Argentina); Carlos M Luna ; Alessio Gerardo Malberti ; Nora Morcillo and Silvina Tartara ; Claudia Pensotti ; Betiana Pereyra ; Pablo Gustavo Scapellato ; Juan Pablo Stagnaro . Australia\u2014Sonali Shah . Austria\u2013Felix L\u00f6tsch, Florian Thalhammer . Belgium\u2014Jean Louis Vincent ; Kurt Anseeuw ; Camille A Francois ; Eva Van Braeckel . Benin\u2014Marcel Zannou Djimon, Jules Bashi, Dodo Roger . Brazil\u2014Simone Aranha Nou\u00e9r . Bulgaria\u2014Peter Chipev, Milena Encheva ; Darina Miteva ; Diana Petkova ; Mbatchou Ngahane Bertrand Hugo . China\u2014Ning Shen ; Jin-fu Xu, . Colombia\u2014 Carlos Andres Bustamante Rico, Ricardo Buitrago ; Fernando Jose Pereira Paternina . Congo\u2014Kayembe Ntumba Jean-Marie . Croatia\u2014Vesna Vladic Carevic ; Marko Jakopovic ; Mateja Jankovic ; Zinka Matkovic ; Ivan Mitrecic . Denmark\u2014Marie-Laure Bouchy Jacobsson ; Anette Bro Christensen ; Uff e Christian HeitmannB\u00f8dtger ; Christian Niels Meyer ; Andreas Vestergaard Jensen, Gertrud Baunb\u00e6k-knudsen, Pelle Trier Petersen and Stine Andersen . Egypt\u2014Ibrahim El-Said Abd El-Wahhab ; Nesreen Elsayed Morsy ; Hanaa Shafi ek ; Eman Sobh . France\u2014 Fabrice Bertrand ; Christian Brun- Buisson ; Etienne de Montmollin ; Muriel Fartoukh ; Jonathan Messika ; Pierre Tattevin . Germany\u2014Michael Dreher ; Martin Kolditz ; Matthias Meisinger ; Mathias W Pletz and Stefan Hagel ; Jan Rupp ; Tom Schaberg ; Marc Spielmanns . Ghana\u2014Beatrice Siaw-Lartey . Greece\u2014Katerina Dimakou ; Dimosthenis Papapetrou ; Evdoxia Tsigou and Dimitrios Ampazis, Agioi Anargiroi Hospital, Kifi ssia, Athens, Greece). India\u2014Mohit Bhatia ; Raja Dhar ; George D\u2019Souza ; Rajiv Garg ; Parvaiz A Koul ; P A Mahesh and B S Jayaraj ; Kiran Vishnu Narayan ; Hirennappa B Udnur and Shashi Bhaskara Krishnamurthy . Iran\u2014Keihan Golshani . Ireland\u2014Vera M Keatings ; Ignacio Martin-Loeches (Multidisciplinary Intensive Care Research Organization (MICRO), St James\u2019s University Hospital, Trinity Centre for Health Sciences Dublin, Ireland). Israel\u2014Yasmin Maor ; Jacob Strahilevitz . Italy\u2014Salvatore Battaglia ; Maria Carrabba ; Piero Ceriana ; Marco Confalonieri ; Antonella d\u2019Arminio Monforte ; Bruno Del Prato ; Marino De Rosa ; Riccardo Fantini ; Giuseppe Fiorentino ; Maria Antonia Gammino ; Francesco Menzella ; Giuseppe Milani ; Stefano Nava ; Gerardo Palmiero ; Roberta Petrino and Barbra Gabrielli ; Paolo Rossi ; Claudio Sorino ; Gundi Steinhilber ; Alessandro Zanforlin . Japan\u2014Kiyoyasu Kurahashi . Lebanon\u2014Zeina Aoun Bacha . Mexico\u2014Daniel Barajas Ugalde ; Omar Ceballos Zu\u00f1iga ; Jos\u00e9 F Villegas . Montenegro\u2014Milic Medenica, Hospital for Lung Diseases\u2014Brezovik, Niksic, Montenegro). Netherlands\u2014E M W van de Garde . Nepal\u2014Deebya Raj Mihsra ; Poojan Shrestha, Oxford University Clinical Research Unit, Patan Hospital, Nepal). New Zealand\u2014Elliott Ridgeon . Nigeria\u2014Babatunde Ishola Awokola ; Ogonna N O Nwankwo ; Adefuye Bolanle Olufunlola ; Segaolu Olumide ; Kingsley N Ukwaja . Pakistan\u2014Muhammad Irfan . Poland\u2014Lukasz Minarowski ; Skoczy\u0144ski Szymon . Portugal\u2014Felipe Froes ; Pedro Leuschner ; Mariana Meireles, Cl\u00e1udia Ferr\u00e3o, Pedro Leuschner and Jo\u00e3o Neves ; Sofi a B Ravara ; Cova da Beira Hospital Center, Covilh\u00e3, Portugal). Moldova\u2014Victoria Brocovschii ; Chesov Ion ; Doina Rusu ; Cristina Toma . Romania\u2014Daniela Chirita . Russia\u2014Alexei Birkun ; Anna Kaluzhenina . Saudi Arabia\u2014Abdullah Almotairi , Riyadh, Saudi Arabia); Zakeya Abdulbaqi Ali Bukhary ; Jameela Edathodu ; Amal Fathy ; Abdullah Mushira Abdulaziz Enani and Nazik Eltayeb Mohamed ; Jawed Ulhadi Memon . Serbia\u2014Nada Bogdanovi\u0107 ; Branislava Milenkovic ; Dragica Pesut . Spain\u2014Luis Border\u00ecas, Respiratoy and Sleep Unit, Hospital San Jorge, Huesca, Spain); Noel Manuel Bordon Garcia ; Hugo Cabello Alarc\u00f3n, Sant Hospital Seu de Urgell, Catalonia, Spain); Catia Cilloniz and Antoni Torres , University of Barcelona, Ciber de Enfermedades Respiratorias (CIBERES), Spain); Vicens Diaz-Brito and Xavier Casas ; Alicia Encabo Gonz\u00e1lez ; Maria Luisa Fern\u00e1ndezAlmira ; Miguel Gallego ; Inmaculada Gaspar-Garc\u00cda ; Juan Gonz\u00e1lez del Castillo ; Patricia Javaloyes Victoria ; Elena Laserna Mart\u00ednez ; Rosa Malo de Molina ; Pedro J Marcos ; Rosario Men\u00e9ndez ; Ana Pando-Sandoval ; Cristina Prat Aymerich, Alicia Lacoma del la Torre, and Ignasi Garc\u00eda-Oliv\u00e9 ; Jordi Rello and Silvia Moyano ; Francisco Sanz ; Oriol Sibila and Ana Rodrigo-Troyano ; Jordi Sol\u00e9-Viol\u00e1n ; Ane Uranga ; Job FM van Boven ; Ester Vendrell Torra and Jordi Almirall Pujol . South Africa\u2014Charles Feldman . South Korea\u2014Ho Kee Yum . Togo\u2014Arnauld Attannon Fiogbe . Tunisia\u2014Ferdaous Yangui , Marsa, Tunis, Tunisia). Turkey\u2014Semra Bilaceroglu ; Levent Dalar ; Ufuk Yilmaz . Ukraine\u2014Artemii Bogomolov . United Arab Emirates\u2014Naheed Elahi ; UK\u2014Devesh J Dhasmana ; Rhiannon Ions, Julie Skeemer, and Gerrit Woltmann ; Carole Hancock ; Adam T Hill ; Banu Rudran ; Silvia Ruiz- Buitrago and Marion Campbell ; Paul Whitaker . USA\u2014Karen S Allen ; Veronica Brito ; Jessica Dietz ; Claire E Dysart and Susan M Kellie ; Ricardo A Franco-Sadud and Garnet Meier ; Mina Gaga ; Thomas L Holland and Stephen P Bergin ; Fayez Kheir ; Mark Landmeier ; Manuel Lois ; Girish B Nair ; Hemali Patel ; Katherine Reyes ; William Rodriguez-Cintron ; Shigeki Saito ; Nilam J Soni, Julio Noda, Cecilia I Hinojosa, Stephanie M Levine, Luis F Angel, and Antonio Anzueto ; K Scott Whitlow, John Hipskind, and Kunal Sukhija ; Richard G. Wunderink and Ray D Shah . Zambia\u2014Kondwelani John Mateyo ."} +{"text": "In the editorial section, Matthew Neilson et al. (170) call for improved quality of care in conflict-affected settings. M Anne Yu et al. (171) explain how WHO is coordinating the deployment of vaccines for COVID-19. In the news section, Tatum Anderson (174\u2013175) reports on barriers and solutions to national vaccine deployments. Claire Wardle talks to Andr\u00e9ia Azevedo Soares (176\u2013177) about the challenges faced in addressing false or misleading information about health.Md Nuruzzaman Khan et al. (201\u2013208) track access to female contraceptives.Jorge C\u00e9sar Correia et al. (209\u2013219) review the evidence on effective delivery of services.Edson Serv\u00e1n-Mori et al. (190\u2013200) assess the continuum of careSonam Vijay et al. (236\u2013238) recount the process for introduction. Tsakane MAG Hlongwane et al. (236\u2013238) study implementation of antenatal care recommendations.Nicola L Boddington et al. (178\u2013189) describe clinical and epidemiological characteristics.Fabr\u00edcio Silveira et al. (228\u2013235) calculate convergence on indicators.Christine \u00c5rdal et al. (239\u2013240) propose using the system for environmental surveillance of polio."} +{"text": "Digitale zorg op afstand in Diabetes richtlijnen.Digitale zorg op afstand heeft de afgelopen jaren een enorme vlucht genomen. Tijdens de coronapandemie hebben pati\u00ebnten en zorgverleners in korte tijd veel ervaring opgedaan met de digitale tools die er nu al zijn. Zorg op afstand lijkt vooral voor de diabeteszorg aantrekkelijk, omdat diabetespati\u00ebnten veel digitaal meten en hun zorg voor een groot deel planbaar is. Technologie stelt de zorgverlener in staat om de zorg op een andere manier te organiseren en persoonsgerichter vorm te geven. Voor de meeste digitale toepassingen is er nog onvoldoende bewijs voor positieve effecten op de kwaliteit van zorg, de pati\u00ebnttevredenheid of uitkomstmaten zoals tijd-en kostenbesparingen. Daardoor zijn er nog nauwelijks aanbevelingen opgenomen in de bestaande richtlijnen en (zorg)standaarden. De behoefte hieraan neemt wel toe. Daarom is recent een start gemaakt met het DiDia-project, Het NHG voert dit project uit in samenwerking met de Nederlandse Internisten Vereniging, het Kennisinstituut van de Federatie Medisch Specialisten, het National eHealth Living Lab, de Pati\u00ebntenfederatie Nederland, Diabetesvereniging Nederland en de Nederlandse Diabetes Federatie. ZonMw financiert het project.Het doel is om concrete aanbevelingen te formuleren over effectieve, persoonsgerichte zorg op maat. Dit varieert wat betreft consult en contact voor volwassenen met diabetes mellitus type 1 en type 2. Hierbij worden recente ervaringen meegenomen van voor en tijdens de coronapandemie, waar mogelijk onderbouwd met beschikbaar wetenschappelijk onderzoek, ervaringen en geleerde lessen. De aanbevelingen worden opgenomen in de landelijke richtlijnen en (zorg)standaarden voor diabetes in de eerste en tweede lijn, in de informatie voor pati\u00ebnten op Thuisarts.nl en in de NDF Toolkit persoonsgerichte diabeteszorg en preventie.Het gebruik van digitale middelen neemt zowel in de eerste als in de tweede lijn toe. Tijdens de coronapandemie is bij zorgverleners met name het gebruik van e-consulten (82% bij huisartsen), beeldbellen (55% bij huisartsen) en pati\u00ebntportalen (55% bij huisartsen) gestegen . Een versnelling in de digitale zorgverlening doet zich ook voor in de tweede lijn, versterkt door continue glucosemetingen. Het is een belangrijke ontwikkeling dat pati\u00ebnten en zorgverleners via bepaalde applicaties kunnen communiceren en meetwaarden direct, realtime kunnen doorgeven.evidence dat dit wisselend wordt ervaren. De e-healthtoe-passingen moeten dan ook aansluiten bij de wensen van pati\u00ebnten en zorgverleners, en de zorg zodanig onder-steunen dat deze van meerwaarde is voor de pati\u00ebnt \u00e9n de zorgverlener.\u2019Prof. dr. Jako Burgers, huisarts en leerstoelhouder van het Nederlands Huisartsen Genootschap: \u2018Heel belangrijk is het dat we de verschillende mogelijkheden goed weergeven in de richtlijnen, met betrouwbare en onafhankelijke informatie over de voor- en de nadelen. De huidige zorg is kwalitatief goed en mag dan ook zeker niet minder worden door zorg op afstand. Voor sommige pati\u00ebnten-groepen kunnen we dankzij de digitale hulpmiddelen betere en toegankelijkere zorg bieden. Voor hen is \u201czorg op afstand\u201d eigenlijk ook \u201czorg dichtbij\u201d, omdat zij minder frequent naar het ziekenhuis hoeven. Dit geldt niet in alle gevallen. We willen dan ook systematisch informatie en ervaringen ophalen en onderbouwde keuzes maken. Neem het videoconsult, ofwel beeldbellen. Dat is veel toegepast tijdens de coronapandemie, maar er is steeds meer Naast de positieve ervaringen zijn er ook knelpunten. Digitale opslag van metingen en gegevens biedt de mogelijkheid om deze direct te koppelen aan het elektronisch pati\u00ebntendossier en/of aan een persoonlijke gezondheids-omgeving (PGO), waardoor telemonitoring mogelijk wordt. Lastig hierbij is de grote variatie in het gebruik van digitale middelen \u2013 zowel bij pati\u00ebnten als bij zorgverleners \u2013 en dat de data die worden ingewonnen via allerlei verschillende platforms toegankelijk zijn. Een groot knelpunt is dat de data hierdoor niet eenvoudig ingevoerd kunnen worden in het HIS. Die platforms zijn veelal gerelateerd aan de producenten van diabeteshulpmiddelen, zoals pomp en sensor. Het proces om tot heldere aanbevelingen te komen, vertraagt hierdoor. Daarnaast voldoen de meeste PGO\u2019s nog niet aan de MedMij-criteria. Bovendien staat de commerci\u00eble markt van software-aan-bieders uniformiteit en samenwerking in de weg.Burgers: \u2018Juist de kanttekeningen willen we ook goed uitzoeken om gericht advies te geven wanneer iets wel bijdraagt aan goede zorg en wanneer niet. Bij het beoordelen van de digitale toepassingen kijken we naar verschillende aspecten: hoe werkt het, hoe betrouwbaar is het, hoe groot is de groep die er voordeel bij kan hebben, hoe staan zorgverleners en pati\u00ebnten ertegenover, wat zijn de voorwaarden en hoe bed je het gebruik op een goede manier in binnen de aan te bieden zorg? Er zijn al veel mogelijkheden en de markt breidt zich snel uit. Dat kan ook meer ruis geven. We willen duidelijke randvoor-waarden formuleren; binnen de toepassingen die daaraan voldoen, is er keuzevrijheid.\u2019eerste fase worden ervaringen met zorg op afstand en implementatie-vraagstukken over zorg op afstand ge\u00efnventariseerd in focusgroepen met pati\u00ebnten en zorgprofessionals uit de eerste en tweede lijn.Om tot gedragen aanbevelingen te komen, is een plan van aanpak opgesteld, dat bestaat uit vijf fases. In de Focusgroepen 1 en 2 bestaan uit mensen met type 1-diabetes en type 2-diabetes uit de eerste (10 pati\u00ebnten met type 1-diabetes) en de tweede lijn (8 pati\u00ebnten met type 1-diabetes en 2 pati\u00ebnten met type 2-diabetes).Focusgroep 3 bestaat uit zorgverleners uit de eerste lijn: 4 huisartsen, 2 kader-huisartsen en 2-3 praktijkonder-steuners en 2 di\u00ebtisten.Focusgroep 4 bestaat uit zorgverleners uit de tweede lijn: 4 internisten, 2 kader-huisartsen, 2-3 diabetesver-pleegkundigen en 2 di\u00ebtisten.Focusgroep 5 bestaat uit overige zorgverleners: 2 podo-therapeuten, 2 apothekers, 2 fysiotherapeuten, 2 kader-huisartsen en 2 praktijkondersteuners.Daarnaast worden kwantitatieve data verzameld, literatuuronderzoek gedaan en wordt een knelpunteninventarisatie verricht.tweede fase worden door middel van een invitational conference ervaringen, knelpunten en bevorderende en belemmerende factoren bij de toepassing van zorg op afstand besproken en geprioriteerd om gezamenlijk tot consensus te komen welke knelpunten in de volgende fase worden meegenomen naar de richtlijnen.In de derde fase gebruikt om aanbevelingen te formuleren. Burgers: \u2018De bestaande werkgroepen van de NHG-Standaard Diabetes mellitus type 2 en de diabetesrichtlijnen van de Nederlandse Internisten Vereniging (NIV) zullen volgens de gebruikelijke methode van richtlijnontwikkeling de uitgangsvragen uitwerken. Zij kijken kritisch naar wat uit de literatuur en uit de ervaringen komt.\u2019De opgedane kennis, inzichten en ervaringen worden in de invitational conference tot een geprioriteerde lijst met aandachtspunten. Deze worden gebruikt bij het opstellen van specifieke aanbevelingen over zorg op afstand in de diabetesrichtlijnen van de eerste en de tweede lijn. Daarnaast leiden deze aandachtspunten tot een overzicht van bevorderende en belemmerende factoren die de basis vormen van een implementatieplan (fase 4), inclusief (rand)voorwaarden waaraan zorg op afstand zou moeten voldoen voor toepassing in de praktijk. In fase 5 wordt het project afgerond met een eindrapportage.De praktijkervaringen, kwantitatieve data, het literatuur-onderzoek en de knelpunten leiden na de Hybride Diabeteszorg zetten wij ons in voor de kwaliteitsslag die nodig is om ervaringen te \u201cverzilveren\u201d die gerelateerd zijn aan de opkomst van smart diabetes devices en die zijn opgedaan tijdens de coronapandemie. Denk bijvoorbeeld aan alternatieve consult-vormen. Daarbij is het idee dat effectieve, persoonsgerichte zorg op maat kan vari\u00ebren wat betreft consult en type contact. Er zijn nog veel vragen die beantwoord moeten worden om kwaliteit te defini\u00ebren en te borgen. Dat kan goed in richtlijnen en we zijn daarom blij hier via DiDia een bijdrage aan te kunnen leveren. We zullen de opbrengsten van het traject opnemen in de NDF Zorgstandaard Diabetes en doorvertalen naar producten voor de NDF Toolkit persoonsgerichte diabeteszorg en preventie.\u2019Het DiDia-project is begin 2022 gestart en heeft een looptijd van achttien maanden. De NDF is zoals aangegeven \u00e9\u00e9n van de partners in het DiDia-traject. Corrine Brinkman, strategisch beleidsadviseur bij de NDF: \u2018Binnen het NDF Programma Burgers: \u2018Ik vind het bijzonder waardevol dat we dit project met zo veel partijen uitvoeren. De coronapandemie heeft naast ellende ook positieve dingen gebracht. De onderlinge samenwerking is verbeterd, tussen zorgprofessionals onderling en tussen zorgprofessionals en pati\u00ebnten. Dat moeten we zeker vasthouden. Ik ben heel enthousiast om specifiek op dit onderwerp, door het inzetten van digitale hulpmiddelen, het aanbod van zorg te vergroten en aan te passen aan de mogelijkheden van deze tijd en daardoor de zorg te verbeteren. Ik zou zorgverleners willen meegeven: wees niet bang om nieuwe digitale toepassingen uit te proberen. Het is niet zo dat de diabeteszorg straks volledig algoritmegestuurd wordt. Persoonsgerichte zorg blijft leidend. De toepassingen zijn er niet ter vervanging, maar ter ondersteuning van besluitvorming.\u2019"} +{"text": "In editorials this month, Muhammad Naveed Noor et al. (682) argue the case for a classification of the private health sector. Sadia Shakoor and Sabine Dittrich (683) point out that better diagnostic tools are needed to distinguish typhoid from other causes of acute febrile illness.Fid Thomson (686\u2013687) reports on the potential of affordable solar power to transform health service delivery in facilities with no electricity. Christian Owoo talks to Gary Humphreys (688\u2013689) about the clinical challenges faced during the COVID-19 pandemic and the importance of adapting guidance to local conditions.Remco Peters et al. (690\u2013706) review national surveillance programmes.Mohamed A Jama et al. (738\u2013742) align stakeholders on improved service delivery.Homa Attar Cohen et al. (707\u2013716) describe ongoing genomic surveillance effortsRamona Ludolph et al. (717\u2013722) present a research agenda.Ana Pilar Betr\u00e1n et al. (723\u2013729) detail the research needed to improve outcomes for mothers and babies. Ankur Gupta-Wright et al. (730\u2013737) list the desired characteristics of new tests.Renzo Bianchi and Irvin Sam Schonfeld (743\u2013745) reexamine the basis of original definitions."} +{"text": "In the editorial section this month, Lisa M Askie et al. (618) call for papers on the impact of WHO\u2019s normative and standard-setting function. Maria Eugenia Grillet (619) discusses what\u2019s needed to sustain malaria elimination in the Region of the Americas.Gary Humphreys and Lynn Eaton (622\u2013623) report on the release of mosquito nets that use chlorfenapyr. Rosanna Peeling talks to Gary Humphreys (624\u2013625) about the need for more use of new diagnostic tools.Dechen Wangmo et al. (679\u2013680) commit to action in mental health.Etienne Rupp\u00e9 et al. (672\u2013678) introduce a surveillance network for antimicrobial resistant pathogens.Ryan K McBain et al (626\u2013636) determine the costs associated with provided HIV treatment.Junjie Hua et al. (637\u2013648) assess coding data quality in WHO\u2019s mortality statistics.Laila Khawar et al. (649\u2013665) review eradication and elimination goals.Michelle R Kaufman et al. (666\u2013671) explain why sex and gender need to be accounted for correctly."} +{"text": "In the editorial section, Irene Papanicolas et al. (438) call for papers for an upcoming theme issue on health system performance assessment. Gary Humphreys (441 \u2013 442) reports on innovative approaches to vector control in the Region of the Americas and speaks to Jackeline Alger (443 \u2013444) about building public health research capacity in Honduras against a backdrop of conflict and political upheaval.Puhong Zhang et al. (453 \u2013 469) quantify sodium levels in pre-packaged foods. Keertana Duppala et al. (445 \u2013 452) test feasibility of simultaneous screening. Tsetsegee Sambuu et al. (470 \u2013 477) detect an increase in domestic cases of carbon monoxide poisoning. Lydia E Pace et al. (478 \u2013 486) trial combining clinical breast examinations with cervical cancer screening.Ho Quang Chanh et al. (487 \u2013 492) share lessons learnt with a prototype wearable device."} +{"text": "We describe demographic features, treatments and clinical outcomes in the International Severe Acute Respiratory and emerging Infection Consortium (ISARIC) COVID-19 cohort, one of the world's largest international, standardized data sets concerning hospitalized patients.The data set analysed includes COVID-19 patients hospitalized between January 2020 and January 2022 in 52 countries. We investigated how symptoms on admission, co-morbidities, risk factors and treatments varied by age, sex and other characteristics. We used Cox regression models to investigate associations between demographics, symptoms, co-morbidities and other factors with risk of death, admission to an intensive care unit (ICU) and invasive mechanical ventilation (IMV).Data were available for 689\u200a572 patients with laboratory-confirmed (91.1%) or clinically diagnosed (8.9%) SARS-CoV-2 infection from 52 countries. Age [adjusted hazard ratio per 10\u2009years 1.49 ] and male sex [1.23 ] were associated with a higher risk of death. Rates of admission to an ICU and use of IMV increased with age up to age 60\u00a0years then dropped. Symptoms, co-morbidities and treatments varied by age and had varied associations with clinical outcomes. The case-fatality ratio varied by country partly due to differences in the clinical characteristics of recruited patients and was on average 21.5%.Age was the strongest determinant of risk of death, with a \u223c30-fold difference between the oldest and youngest groups; each of the co-morbidities included was associated with up to an almost 2-fold increase in risk. Smoking and obesity were also associated with a higher risk of death.\u00a0The size of our international database and the standardized data collection method make this study a comprehensive international description of COVID-19 clinical features. Our findings may inform strategies that involve prioritization of patients hospitalized with COVID-19 who have a higher risk of death. Key MessagesSeveral studies have investigated the risks of severe illness and death due to infection with SARS-CoV-2, providing estimates of the case-fatality ratio in different settings and risk factors for these outcomes, but these tended to be national studies conducted over a short time period.We show how clinical presentation and risks of death and admission to an intensive care unit vary with patient characteristics based on a very large number of patient records from 52 countries, collected using standardized data collection tools.Age was the strongest determinant of risk; pre-existing co-morbidities and male sex were also associated with higher risk of death.Smoking and obesity are modifiable risk factors that are associated with a higher risk of death.Our findings may inform strategies that involve prioritization of patients, globally, who have a higher risk of adverse outcomes if hospitalized with COVID-19, as well as prevention strategies.To respond to COVID-19, policymakers and clinicians need robust data to drive the decision-making processes that save or cost lives. Observational cohort data describing clinical characteristics and the likelihood of severe outcomes can guide health policy development, produce research hypotheses for clinical trials and improve clinical guidelines for patient care.,The Clinical Characterisation Protocol (CCP) developed by the International Severe Acute Respiratory and emerging Infection Consortium (ISARIC) and the World Health Organization (WHO)We present data from an international cohort of almost 700\u200a000 patients from 1380 sites across 52 countries. We summarize the demographic features and clinical presentation of hospitalized patients with COVID-19 in low-, middle- and high-resource settings. We characterize the variability in clinical features in these patients and explore the risk factors associated with mortality, and the need for intensive care and mechanical ventilation, on a global scale. We aimed to report a general description of this international data set 2\u00a0years after the beginning of the pandemic, to estimate the frequency of co-morbidities, risk factors, symptoms and use of different treatments, and to estimate the risk of severe clinical outcomes and the associations of various factors with risk of these outcomes.IJE online) before uploading to the central data repository. Centrally collated data were wrangled and mapped to the structure and controlled terminologies of Study Data Tabulation Model using Trifacta\u00ae software. The data collection, aggregation, curation and harmonization process has been previously described.We used international prospective observational data of clinical features and outcomes of patients admitted to hospital with COVID-19. The ISARIC/WHO CCP, incorporating Short PeRiod IncideNce sTudy of Severe Acute Respiratory Infection (SPRINT-SARI),We included hospitalized patients of any age with clinically or laboratory-diagnosed COVID-19. This analysis included patients admitted to hospital in any of the countries which contributed data. It also includes a small subset of asymptomatic patients who were admitted to the hospital purely for isolation (<1%).IJE online). We calculated case-fatality ratios (CFRs) overall, by country and by month, using the method suggested by Ghani et al.We excluded patients with missing age or sex from all analyses . AdditioPatients were followed up from hospital admission to death, discharge or censoring, whichever occurred first. Cox proportional hazards models were used to assess the association of demographic variables, co-morbidities and symptoms at admission (unless symptom onset was after admission) with risk of death, admission to an intensive care unit (ICU) or a high-dependency unit (hereafter referred to collectively as ICU) or use of invasive mechanical ventilation (IMV). Individuals were censored if they were lost to follow-up (e.g. transferred to another facility) or remained in hospital on 6 January 2022. Time from symptom onset to the time of death or censoring, whichever occurred earlier, was used as the timescale. Patients were considered at risk from the time of symptom onset or admission, whichever occurred later. For all outcomes, censoring times of discharged patients were modified and set to be equal to the maximum time to censoring/event (to account for informative censoring). For associations with admission to an ICU, the timescale was from symptom onset to the earliest of admission to an ICU, death, discharge and censoring. The event was admission to an ICU. For associations with receipt of IMV, the timescale was from symptom onset to the earliest of IMV, death, discharge and censoring. The event was IMV. Models were adjusted for age and sex, and stratified by country. We grouped countries with <50 individuals into a single category. Hazard ratios (HRs) and 95% CIs were estimated. We assessed the proportional hazards assumption using scaled Schoenfeld residuals. For explanatory variables with multiple categories (such as age groups), we used quasi-standard errorsWe repeated the main analyses for patients with laboratory-confirmed SARS-CoV-2 only, as a sensitivity analysis. For associations of age and sex with risk of death, we also estimated HRs within each country and calculated an overall HR using inverse-variance weighting. For age, sex and co-morbidities, we also estimated associations within the first (2020) and second year (2021) of the pandemic.In total, 9442 individuals had missing age or sex and weresurvival, ggplot2, qvcalc and finalfit.Analysis was performed using R version 4.1.1 and packages This was an urgent public health research study in response to a Public Health Emergency of International Concern. Patients or the public were not involved in the design, conduct or reporting of this rapid response research. ISARIC, ISARIC4C, the National Institute of Communicable Diseases South Africa and other collaborators have public facing websites and social media accounts to disseminate findings. The contributing individuals and institutions engage with print and internet press, television, radio, news and documentary programme makers to share evidence with the public and invite feedback.IJE online). Overall, 91.1% of the participants included in the primary analysis had a positive SARS-CoV-2 PCR test . In particular there were differences in age, region and vital signs (heart and respiratory rate and blood pressure). Anthropometric variables and vital signs varied by age (IJE online).The majority of patients were recruited in South Africa (59.3%) and 31.1% in the UK . ParticiIJE online). There was some variation by age and country (IJE online). Fatigue/malaise, cough and shortness of breath were most prevalent amongst patients who were 40\u201370\u2009years old. The prevalence of altered consciousness/confusion increased with age and was reported in 28.4% of patients of >80\u2009years of age. Loss or altered smell or taste were not commonly reported but there was a high proportion of missing values for these two symptoms (39.3% for loss of smell and 40.5% for taste). We have previously described the associations of age and gender with presenting symptoms.IJE online) but there were more missing values among individuals with only a clinical diagnosis (IJE online). Altered consciousness/confusion, cough, fatigue/malaise, fever, shortness of breath and vomiting/nausea were more frequently reported in patients with laboratory-confirmed SARS-CoV-2 infection than in those with a clinical diagnosis alone (IJE online).The most common symptoms on presentation were shortness of breath (50%), cough (48.5%) and fever (44.3%) (IJE online). Individuals aged 40\u201370\u2009years were more likely to meet each of the four case definitions based on symptoms than patients at the extremes of the age distribution (IJE online).Overall, 50\u201370% of patients met each of the international symptom-based case definitions. These proportions were higher among those with laboratory-confirmed SARS-CoV-2 infection (ribution . Adults IJE online). The median white blood cell count (7.2IJE online).Routine blood test results are shown in\u00a0IJE online). Among 538\u200a974 individuals with data available for any five or more co-morbidities or risk factors, 165\u200a987 (30.8%) had no co-morbidities reported. The prevalence of most co-morbidities varied by age (IJE online). The prevalence of chronic cardiac disease, chronic kidney disease, dementia, hypertension and rheumatologic disorder increased with age. The prevalence of diabetes was highest in individuals aged 60\u201380\u00a0years. There were 26\u200a776 patients with HIV infection, 11\u200a384 with tuberculosis and 5044 with both; 25\u200a368 of the patients with HIV infection and 11\u200a137 patients with tuberculosis were from South Africa. Obesity was reported for 48\u200a077 participants, smoking for 65\u200a056 and pregnancy for 18\u200a669.The most common pre-existing co-morbidities were hypertension, diabetes and chronic cardiac disease , 179\u200a448 (65.2%) received oxygen therapy at any time during hospitalization, which was delivered via a high-flow nasal cannula to 55\u200a541 (19.8%), by non-invasive ventilation (NIV) to 44\u200a280 (15.8%) and IMV to 38\u200a440 (13.7%) . Type ofIJE online). The proportion of patients receiving antibacterial agents increased with age, as did the proportion receiving corticosteroids up to ages 70\u201380\u00a0years (IJE online).\u00a0Information on antibacterial treatment was available for 255\u200a031 patients, 198\u200a295 (77.8%) of whom received antibacterial agents; 126\u200a391 of 262\u200a385 of patients with data available (48.2%) received corticosteroids. The use of corticosteroids increased after the publication of results of the RECOVERY trialIJE online), in particular among patients who received oxygen supplementation, in line with the trial results.The most used treatments were oxygen therapy, antibacterial agents and corticosteroids (IJE online). Admission criteria likely varied by country and time, contributing to the heterogeneity in illness severity. Death and discharge rates increased over the first 40\u2009days from hospital admission (or symptom onset if this occurred after admission) (IJE online).The CFR varied by country , likely IJE online). Males had a significantly higher risk of death than females, with an HR of 1.23 , adjusting for age (in 10-year groups) and stratifying by country. There was evidence of deviation from the proportional hazards assumption for both variables. There was no particular trend over follow-up for the magnitude of the association of age with death, but the magnitude of the association of male sex with death appeared to increase with increasing time from admission (or symptom onset for patients who developed symptoms after admission). A model stratified by sex was fitted to estimate associations of age with death taking this time-varying association of sex into account. HRs for age estimated by the two models were very similar. HRs were also estimated by sex and age, stratifying by country (IJE online) and sex (IJE online) varied by country. Country-specific HRs for a 10-year increase in age varied from 1.15 to 3.50 , with a pooled estimate of 1.49 , which is the same as the estimate from the main analysis. For sex , HRs were between 0.61 and 3.57 , with a weighted average of 1.26 [compared with 1.23 in the main analysis]. When fitting a model including age (continuous) and sex, and stratified by country, within each year of the pandemic, HRs (95% CI) per 10\u2009years of higher age were 1.50 for 2020 and 1.47 for 2021, and for males vs females 1.31 and 1.20 , respectively.The risk of death was higher for males than for females . Older a country . HRs for2 and sex, and stratifying by country] and tuberculosis [1.56 ] were associated with the highest relative risks of death; there was no evidence of an interaction (P\u2009=\u20090.18). Since many patients with these co-morbidities were from South Africa, we assessed the associations separately among patients from South Africa and patients from all other countries by fitting a model including HIV, tuberculosis, age and age stratified by sex and country. The associations with risk of death did not significantly differ between patients from South Africa and patients from other countries, although estimates were numerically higher for patients from South Africa: respectively for HIV HR (95% CI) 1.52 and 1.18 and for tuberculosis 1.35 and 1.18 . All reported co-morbidities were associated with a higher risk of death, except rheumatologic disorder and asthma for which there was no evidence of an association (IJE online). Obesity was associated with a higher risk of death [HR 1.24 ]. Smoking was also associated with a higher risk of death [1.10 ]. Pregnancy was associated with a lower risk of death . There were small differences in the magnitude of associations of co-morbidities with risk of death between calendar years (IJE online).Overall, HIV infection , shortness of breath [1.76 ], lymphadenopathy [1.45 ], wheezing [1.40 ], severe dehydration [1.36 ], altered consciousness/confusion [1.32 ], skin rash [1.30 ], cough [1.16 ], fever [1.18 ] and fatigue/malaise [1.06 ] were associated with a higher risk of death (IJE online). In general, gastrointestinal, musculoskeletal symptoms and loss of or altered taste or smell were associated with a lower risk of dying; e.g. nausea/vomiting had a HR of 0.81 , abdominal pain 0.84 , diarrhoea 0.93 and muscle aches/joint pain 0.88 .Inability to walk [HR 1.84 , adjusting for age, ageIJE online).The risk of admission to an ICU increased with age after age 30\u00a0years and started decreasing from age 60\u00a0years, with patients aged >80\u00a0years being unlikely to be admitted to an ICU. Men were more likely to be admitted to an ICU overall, with a HR of 1.17 . There was evidence of non-proportional hazards, indicating that the relative risk changed with time since symptom onset . There were similar patterns for risk of IMV . There wThe ISARIC international cohort study included at the time of these analyses standardized data on almost 700\u200a000 patients from 1380 sites across 52 countries. To our knowledge, this is the most extensive in-hospital COVID-19 cohort study in the world. The size and breadth of the study allow us to evaluate the contribution of individual risk factors to outcomes such as death, admission to an ICU and use of mechanical ventilation. The value of the international cohort design is its capacity to cover the breadth of COVID-19 characteristics unencumbered by differences in classification and reporting. Furthermore, our international cohort design allowed us to explore risk factors that are globally uncommon, or uncommon in cohorts from high-income countries. For example, our data set is the largest prospective cohort study of COVID-19 patients with HIV infection, tuberculosis, malnutrition, pregnancy and transplantation. Although the study population includes children and we have presented some of their characteristics, a detailed analysis of the children population is beyond the scope of this paper.Across the cohort, the most common presenting symptoms were fever, shortness of breath and cough. Among other symptoms reported, the most common were altered consciousness in older patients and gastrointestinal symptoms in younger patients. Our data show that about one-third of patients do not meet one of the four most widely used case definitions at the time of hospitalization, particularly those in the younger and older age groups. These differences are relevant when defining testing or isolation and for early detection of new clusters and variants. Although case definitions must be simple, age-specific definitions may improve sensitivity. This has implications also for case management; about one-third of patients did not require any oxygen therapy during their hospitalization.,Our study confirms that the strong association between age and risk of death from COVID-19 is a global phenomenon. The elderly are at a significantly higher risk of death from COVID-19. Every decade of life adds a 50% risk of dying, with those aged >90\u00a0years having a 17-fold higher risk than 20- to 30-year-olds. Although similar results were shown globally for non-hospitalized cases,,,,We found five co-morbidities to be strongly associated with risk of death. The most substantial risk factor was HIV infection. There was a high proportion of people living with HIV (PLWH) in this cohort. Whilst retrospective health records analyses have been performed previously,Globally, CFRs were much higher in the 5% of patients who were admitted to an ICU on the first day of their admission than those who required an ICU later during their admission. The risk of admission to an ICU increased with age, but then started decreasing from age 60\u2009years, with patients aged >80\u2009years being very unlikely to be admitted to an ICU. It has been suggested that patients who have severe illness and are not treated in an ICU have poorer outcomes.This international cohort study overcomes some of the traditional problems of multicentre observational studies by using standardized variables and outcome measures. Our data are likely to be of value in modelling and health system planning. For example, we note the greatly increased risk of death amongst patients with tuberculosis and malnutrition in our cohort and protecting such individuals from COVID-19 must be a critical public health priority for countries with high prevalence rates of these conditions. Equally concerning is our finding of increased risk of death amongst PLWH. Many PLWH reside in sub-Saharan Africa and our data may indicate a phenomenon that is currently hidden due to under-testing for SARS-CoV-2Whereas our study includes a broad range of data from different countries, various sites have different levels of data completeness. For example, we cannot evaluate the proportion of patients with HIV infection or tuberculosis who were taking appropriate, effective treatments. We have no further detail on the type of organ received by the transplantation cohort. Missing data may have affected the estimates of prevalence of symptoms, co-morbidities and other patient characteristics, as well as estimates of associations between these and risks of outcomes. We presented missing data in each variable in tables and figures. Specifically for symptoms and treatments, data from South Africa were not available and aggregated estimates might largely reflect data from the UK. Whereas we have produced a summary of the associations of risk factors with outcomes in COVID-19, pandemics are complex, dynamic phenomena. There is variation in the amount and completeness of data between countries and between sites within each country. Although we have adjusted for potential confounders, inclusion of patients in the data varies by country/site, which may lead to bias in estimates of association and of absolute risks such as CFR. Our findings will increasingly be influenced by the provision of vaccination, which we have not examined in this study, and effective treatments, as well as the variability in access to these measures in the global context. We do not include data on SARS-CoV-2 variants of concern in this paper. The majority of submitted case records come from two countries: the UK and South Africa; however, there were 22 countries with data on >500 patients.This paper represents the largest international cohort of hospitalized COVID-19 patients published to date. We demonstrate several associations of global importance, including an increased risk of death in patients with HIV and tuberculosis. Co-morbidities were associated with a higher risk of death, each associated with up to a 2-fold increase. Smoking and obesity were also associated with a higher risk of death. Age was most strongly associated with risk of death, with a \u223c30-fold difference between the oldest and youngest groups. These findings may be used to inform strategies that involve prioritization of high-risk patients hospitalized with COVID-19 and prevention strategies. The ISARIC global collaboration continues to collect standardized data which will enable continued data-led comparisons as the world implements vaccination, treatment and public health control strategies.ISARIC Clinical Characterisation Group: Ali Abbas; Sheryl Ann Abdukahil; Nurul Najmee Abdulkadir; Ryuzo Abe; Laurent Abel; Lara Absil; Subhash Acharya; Andrew Acker; Elisabeth Adam; Diana Adri\u00e3o; Saleh Al Ageel; Shakeel Ahmed; Kate Ainscough; Eka Airlangga; Tharwat Aisa; Ali Ait Hssain; Younes Ait Tamlihat; Takako Akimoto; Ernita Akmal; Eman Al Qasim; Razi Alalqam; Angela Alberti; Tala Al-dabbous; Senthilkumar Alegesan; Cynthia Alegre; Marta Alessi; Beatrice Alex; K\u00e9vin Alexandre; Abdulrahman Al-Fares; Huda Alfoudri; Imran Ali; Adam Ali; Naseem Ali Shah; Kazali Enagnon Alidjnou; Jeffrey Aliudin; Qabas Alkhafajee; Clotilde Allavena; Nathalie Allou; Aneela Altaf; Jo\u00e3o Alves; Rita Alves; Jo\u00e3o Melo Alves; Maria Amaral; Nur Amira; Phoebe Ampaw; Roberto Andini; Claire Andr\u00e9jak; Andrea Angheben; Fran\u00e7ois Angoulvant; S\u00e9verine Ansart; Sivanesen Anthonidass; Massimo Antonelli; Carlos Alexandre Antunes de Brito; Ardiyan Apriyana; Yaseen Arabi; Irene Aragao; Francisco Arancibia; Carolline Araujo; Antonio Arcadipane; Patrick Archambault; Lukas Arenz; Jean-Beno\u00eet Arlet; Christel Arnold-Day; Lovkesh Arora; Rakesh Arora; Elise Artaud-Macari; Diptesh Aryal; Angel Asensio; Muhammad Ashraf; Namra Asif; Mohammad Asim; Jean Baptiste Assie; Amirul Asyraf; Anika Atique; AM Udara Lakshan Attanyake; Johann Auchabie; Hugues Aumaitre; Adrien Auvet; Laur\u00e8ne Azemar; Cecile Azoulay; Benjamin Bach; Delphine Bachelet; Claudine Badr; Nadia Baig; J Kevin Baird; Erica Bak; Agamemnon Bakakos; Nazreen Abu Bakar; Andriy Bal; Mohanaprasanth Balakrishnan; Valeria Balan; Firouz\u00e9 Bani-Sadr; Renata Barbalho; Nicholas Yuri Barbosa; Wendy S. Barclay; Saef Umar Barnett; Michaela Barnikel; Helena Barrasa; Audrey Barrelet; Cleide Barrigoto; Marie Bartoli; Mustehan Bashir; Romain Basmaci; Muhammad Fadhli Hassin Basri; Denise Battaglini; Jules Bauer; Diego Fernando Bautista Rincon; Denisse Bazan Dow; Alexandra Bedossa; Ker Hong Bee; Husna Begum; Sylvie Behilill; Albertus Beishuizen; Aleksandr Beljantsev; David Bellemare; Anna Beltrame; Beatriz Amorim Beltr\u00e3o; Marine Beluze; Nicolas Benech; Lionel Eric Benjiman; Dehbia Benkerrou; Suzanne Bennett; Lu\u00eds Bento; Jan-Erik Berdal; Delphine Bergeaud; Hazel Bergin; Jos\u00e9 Luis Bernal Sobrino; Giulia Bertoli; Lorenzo Bertolino; Simon Bessis; Sybille Bevilcaqua; Karine Bezulier; Amar Bhatt; Krishna Bhavsar; Claudia Bianco; Farah Nadiah Bidin; Moirangthem Bikram Singh; Felwa Bin Humaid; Mohd Nazlin Bin Kamarudin; Fran\u00e7ois Bissuel; Patrick Biston; Laurent Bitker; Jonathan Bitton; Pablo Blanco-Schweizer; Catherine Blier; Frank Bloos; Mathieu Blot; Filomena Boccia; Laetitia Bodenes; Alice Bogaarts; Debby Bogaert; Anne-H\u00e9l\u00e8ne Boivin; Pierre-Adrien Bolze; Fran\u00e7ois Bompart; Aurelius Bonfasius; Diogo Borges; Rapha\u00ebl Borie; Hans Martin Bosse; Elisabeth Botelho-Nevers; Lila Bouadma; Olivier Bouchaud; Sabelline Bouchez; Dounia Bouhmani; Damien Bouhour; K\u00e9vin Bouiller; Laurence Bouillet; Camile Bouisse; Anne-Sophie Boureau; John Bourke; Maude Bouscambert; Aurore Bousquet; Jason Bouziotis; Bianca Boxma; Marielle Boyer-Besseyre; Maria Boylan; Axelle Braconnier; Cynthia Braga; Timo Brandenburger; Filipa Br\u00e1s Monteiro; Luca Brazzi; Patrick Breen; Dorothy Breen; Patrick Breen; Kathy Brickell; Shaunagh Browne; Alex Browne; Nicolas Brozzi; Marjolein Brusse-Keizer; Nina Buchtele; Christian Buesaquillo; Polina Bugaeva; Marielle Buisson; Danilo Buonsenso; Erlina Burhan; Ingrid G. Bustos; Denis Butnaru; Andr\u00e9 Cabie; Susana Cabral; Eder Caceres; Cyril Cadoz; Mia Callahan; Kate Calligy; Jose Andres Calvache; Jo\u00e3o Cam\u00f5es; Valentine Campana; Paul Campbell; Josie Campisi; Cecilia Canepa; Mireia Cantero; Pauline Caraux-Paz; Sheila C\u00e1rcel; Chiara Simona Cardellino; Sofia Cardoso; Filipe Cardoso; Filipa Cardoso; Nelson Cardoso; Simone Carelli; Nicolas Carlier; Thierry Carmoi; Gayle Carney; In\u00eas Carqueja; Marie-Christine Carret; Fran\u00e7ois Martin Carrier; Ida Carroll; Maire-Laure Casanova; Mariana Casc\u00e3o; Siobhan Casey; Jos\u00e9 Casimiro; Bailey Cassandra; Silvia Casta\u00f1eda; Nidyanara Castanheira; Guylaine Castor-Alexandre; Henry Castrill\u00f3n; Ivo Castro; Ana Catarino; Fran\u00e7ois-Xavier Catherine; Paolo Cattaneo; Roberta Cavalin; Giulio Giovanni Cavalli; Alexandros Cavayas; Adrian Ceccato; Minerva Cervantes-Gonzalez; Anissa Chair; Catherine Chakveatze; Adrienne Chan; Meera Chand; Christelle Chantalat Auger; Jean-Marc Chapplain; Julie Chas; Allegra Chatterjee; Mobin Chaudry; Jonathan Samuel Ch\u00e1vez I\u00f1iguez; Anjellica Chen; Yih-Sharng Chen; Matthew Pellan Cheng; Antoine Cheret; Thibault Chiarabini; Julian Chica; Suresh Kumar Chidambaram; Leong Chin Tho; Catherine Chirouze; Davide Chiumello; Sung-Min Cho; Bernard Cholley; Marie-Charlotte Chopin; Ting Soo Chow; Yock Ping Chow; Hiu Jian Chua; Jonathan Chua; Jose Pedro Cidade; Jos\u00e9 Miguel Cisneros Herreros; Anna Ciullo; Jennifer Clarke; Emma Clarke; Rolando Claure-Del Granado; Sara Clohisey; Perren J. Cobb; Cassidy Codan; Caitriona Cody; Alexandra Coelho; Megan Coles; Gwenha\u00ebl Colin; Michael Collins; Sebastiano Maria Colombo; Pamela Combs; Marie Connor; Anne Conrad; Sof\u00eda Contreras; Elaine Conway; Graham S. Cooke; Mary Copland; Hugues Cordel; Amanda Corley; Sabine Cornelis; Alexander Daniel Cornet; Arianne Joy Corpuz; Andrea Cortegiani; Gr\u00e9gory Corvaisier; Emma Costigan; Camille Couffignal; Sandrine Couffin-Cadiergues; Roxane Courtois; St\u00e9phanie Cousse; Rachel Cregan; Charles Crepy D'Orleans; Cosimo Cristella; Sabine Croonen; Gloria Crowl; Jonathan Crump; Claudina Cruz; Juan Luis Cruz Berm; Jaime Cruz Rojo; Marc Csete; Ailbhe Cullen; Matthew Cummings; Gerard Curley; Elodie Curlier; Colleen Curran; Paula Custodio; Ana da Silva Filipe; Charlene Da Silveira; Al-Awwab Dabaliz; Darren Dahly; Heidi Dalton; Jo Dalton; Seamus Daly; Nick Daneman; Corinne Daniel; Jorge Dantas; Fr\u00e9d\u00e9rick D'Aragon; Menno de Jong; Gillian de Loughry; Diego de Mendoza; Etienne De Montmollin; Rafael Freitas de Oliveira Fran\u00e7a; Ana Isabel de Pinho Oliveira; Rosanna De Rosa; Cristina De Rose; Thushan de Silva; Peter de Vries; Jillian Deacon; David Dean; Alexa Debard; Marie-Pierre Debray; Nathalie DeCastro; William Dechert; Lauren Deconninck; Romain Decours; Eve Defous; Isabelle Delacroix; Eric Delaveuve; Karen Delavigne; Nathalie M Delfos; Ionna Deligiannis; Andrea Dell'Amore; Christelle Delmas; Pierre Delobel; Corine Delsing; Elisa Demonchy; Emmanuelle Denis; Dominique Deplanque; Pieter Depuydt; Mehul Desai; Diane Descamps; Mathilde Desvall\u00e9es; Santi Dewayanti; Pathik Dhanger; Alpha Diallo; Sylvain Diamantis; Andr\u00e9 Dias; Juan Jose Diaz; Priscila Diaz; Rodrigo Diaz; K\u00e9vin Didier; Jean-Luc Diehl; Wim Dieperink; J\u00e9r\u00f4me Dimet; Vincent Dinot; Fara Diop; Alphonsine Diouf; Yael Dishon; F\u00e9lix Djossou; Annemarie B. Docherty; Helen Doherty; Arjen M Dondorp; Andy Dong; Maria Donnelly; Sean Donohue; Yoann Donohue; Chloe Donohue; Peter Doran; C\u00e9line Dorival; Eric D'Ortenzio; James Joshua Douglas; Renee Douma; Nathalie Dournon; Triona Downer; Joanne Downey; Mark Downing; Tom Drake; Aoife Driscoll; Murray Dryden; Murray Dryden; Claudio Duarte Fonseca; Vincent Dubee; Fran\u00e7ois Dubos; Alexandre Ducancelle; Toni Duculan; Susanne Dudman; Abhijit Duggal; Paul Dunand; Mathilde Duplaix; Emanuele Durante-Mangoni; Lucian Durham III; Bertrand Dussol; Juliette Duthoit; Xavier Duval; Anne Margarita Dyrhol-Riise; Sim Choon Ean; Marco Echeverria-Villalobos; Siobhan Egan; Carla Eira; Mohammed El Sanharawi; Subbarao Elapavaluru; Brigitte Elharrar; Jacobien Ellerbroek; Philippine Eloy; Tarek Elshazly; Iqbal Elyazar; Isabelle Enderle; Tomoyuki Endo; Chan Chee Eng; Ilka Engelmann; Vincent Enouf; Olivier Epaulard; Mariano Esperatti; H\u00e9l\u00e8ne Esperou; Marina Esposito-Farese; Jo\u00e3o Estev\u00e3o; Long COVID India Etienne; Manuel Etienne; Nadia Ettalhaoui; Anna Greti Everding; Mirjam Evers; Marc Fabre; Isabelle Fabre; Amna Faheem; Arabella Fahy; Cameron J. Fairfield; Zul Fakar; Komal Fareed; Pedro Faria; Ahmed Farooq; Hanan Fateena; Arie Zainul Fatoni; Karine Faure; Rapha\u00ebl Favory; Mohamed Fayed; Niamh Feely; Laura Feeney; Jorge Fernandes; Mar\u00edlia Andreia Fernandes; Susana Fernandes; Fran\u00e7ois-Xavier Ferrand; Eglantine Ferrand Devouge; Joana Ferr\u00e3o; M\u00e1rio Ferraz; S\u00edlvia Ferreira; Isabel Ferreira; Benigno Ferreira; Ricard Ferrer-Roca; Nicolas Ferriere; C\u00e9line Ficko; Claudia Figueiredo-Mello; William Finlayson; Juan Fiorda; Thomas Flament; Clara Flateau; Tom Fletcher; Letizia Lucia Florio; Deirdre Flynn; Claire Foley; Jean Foley; Victor Fomin; Tatiana Fonseca; Patricia Fontela; Simon Forsyth; Denise Foster; Giuseppe Foti; Erwan Fourn; Robert A. Fowler; Marianne Fraher; Diego Franch-Llasat; John F Fraser; Christophe Fraser; Marcela Vieira Freire; Ana Freitas Ribeiro; Caren Friedrich; Ricardo Fritz; St\u00e9phanie Fry; Nora Fuentes; Masahiro Fukuda; Argin G; Val\u00e9rie Gaborieau; Rostane Gaci; Massimo Gagliardi; Jean-Charles Gagnard; Amandine Gagneux-Brunon; S\u00e9rgio Gai\u00e3o; Linda Gail Skeie; Phil Gallagher; Carrol Gamble; Yasmin Gani; Arthur Garan; Rebekha Garcia; Julia Garcia-Diaz; Esteban Garcia-Gallo; Navya Garimella; Denis Garot; Val\u00e9rie Garrait; Basanta Gauli; Nathalie Gault; Aisling Gavin; Anatoliy Gavrylov; Alexandre Gaymard; Johannes Gebauer; Eva Geraud; Louis Gerbaud Morlaes; Nuno Germano; praveen kumar ghisulal; Jade Ghosn; Marco Giani; Jess Gibson; Tristan Gigante; Morgane Gilg; Elaine Gilroy; Guillermo Giordano; Michelle Girvan; Val\u00e9rie Gissot; Daniel Glikman; Petr Glybochko; Eric Gnall; Geraldine Goco; Fran\u00e7ois Goehringer; Siri Goepel; Jin Yi Goh; Jonathan Golob; Rui Gomes; Kyle Gomez; Joan G\u00f3mez-Junyent; Marie Gominet; Alicia Gonzalez; Patricia Gordon; Isabelle Gorenne; Laure Goubert; C\u00e9cile Goujard; Tiphaine Goulenok; Margarite Grable; Jeronimo Graf; Edward Wilson Grandin; Pascal Granier; Giacomo Grasselli; Christopher A. Green; Courtney Greene; William Greenhalf; Segol\u00e8ne Greffe; Domenico Luca Grieco; Matthew Griffee; Fiona Griffiths; Ioana Grigoras; Albert Groenendijk; Anja Grosse Lordemann; Heidi Gruner; Yusing Gu; J\u00e9r\u00e9mie Guedj; Martin Guego; Dewi Guellec; Anne-Marie Guerguerian; Daniela Guerreiro; Romain Guery; Anne Guillaumot; Laurent Guilleminault; Maisa Guimar\u00e3es de Castro; Thomas Guimard; Marieke Haalboom; Daniel Haber; Hannah Habraken; Ali Hachemi; Amy Hackmann; Nadir Hadri; Fakhir Haidri; Sheeba Hakak; Adam Hall; Sophie Halpin; Jawad Hameed; Ansley Hamer; Raph L. Hamers; Rebecca Hamidfar; Terese Hammond; Lim Yuen Han; Rashan Haniffa; Kok Wei Hao; Hayley Hardwick; Ewen M Harrison; Janet Harrison; Samuel Bernard Ekow Harrison; Alan Hartman; Mohd Shahnaz Hasan; Junaid Hashmi; Muhammad Hayat; Ailbhe Hayes; Leanne Hays; Jan Heerman; Lars Heggelund; Ross Hendry; Martina Hennessy; Aquiles Henriquez-Trujillo; Maxime Hentzien; Jaime Hernandez-Montfort; Andrew Hershey; Liv Hesstvedt; Astarini Hidayah; Eibhilin Higgins; Dawn Higgins; Rupert Higgins; Rita Hinchion; Samuel Hinton; Hiroaki Hiraiwa; Haider Hirkani; Hikombo Hitoto; Yi Bin Ho; Alexandre Hoctin; Isabelle Hoffmann; Wei Han Hoh; Oscar Hoiting; Rebecca Holt; Jan Cato Holter; Juan Pablo Horcajada; Koji Hoshino; Ikram Houas; Catherine L. Hough; Stuart Houltham; Jimmy Ming-Yang Hsu; Jean-S\u00e9bastien Hulot; Stella Huo; Abby Hurd; Iqbal Hussain; Samreen Ijaz; Hajnal-Gabriela Illes; Patrick Imbert; Mohammad Imran; Rana Imran Sikander; Aftab Imtiaz; Hugo In\u00e1cio; Carmen Infante Dominguez; Yun Sii Ing; Elias Iosifidis; Mariachiara Ippolito; Sarah Isgett; Tiago Isidoro; Nadiah Ismail; Margaux Isnard; Junji Itai; Daniel Ivulich; Danielle Jaafar; Salma Jaafoura; Julien Jabot; Clare Jackson; Nina Jamieson; Victoria Janes; Pierre Jaquet; Coline Jaud-Fischer; St\u00e9phane Jaureguiberry; Jeffrey Javidfar; Denise Jaworsky; Florence Jego; Anilawati Mat Jelani; Synne Jenum; Ruth Jimbo-Sotomayor; Ong Yiaw Joe; Ruth N. Jorge Garc\u00eda; C\u00e9dric Joseph; Mark Joseph; Swosti Joshi; Merc\u00e9 Jourdain; Philippe Jouvet; Hanna Jung; Anna Jung; Dafsah Juzar; Ouifiya Kafif; Florentia Kaguelidou; Neerusha Kaisbain; Thavamany Kaleesvran; Sabina Kali; Alina Kalicinska; Smaragdi Kalomoiri; Muhammad Aisar Ayadi Kamaluddin; Zul Amali Che Kamaruddin; Nadiah Kamarudin; Paul Kambiya; Kavita Kamineni; Darshana Hewa Kandamby; Chris Kandel; Kong Yeow Kang; Darakhshan Kanwal; Pratap Karpayah; Todd Karsies; Daisuke Kasugai; Anant Kataria; Kevin Katz; Aasmine Kaur; Christy Kay; Hannah Keane; Se\u00e1n Keating; Pulak Kedia; Claire Kelly; Yvelynne Kelly; Andrea Kelly; Niamh Kelly; Aoife Kelly; Sadie Kelly; Maeve Kelsey; Ryan Kennedy; Kalynn Kennon; Maeve Kernan; Younes Kerroumi; Sharma Keshav; Imrana Khalid; Osama Khalid; Antoine Khalil; Coralie Khan; Irfan Khan; Quratul Ain Khan; Sushil Khanal; Abid Khatak; Amin Khawaja; Krish Kherajani; Michelle E Kho; Saye Khoo; Ryan Khoo; Denisa Khoo; Nasir Khoso; Khor How Kiat; Yuri Kida; Peter Kiiza; Beathe Kiland Granerud; Anders Benjamin Kildal; Jae Burm Kim; Antoine Kimmoun; Detlef Kindgen-Milles; Alexander King; Nobuya Kitamura; Paul Klenerman; Rob Klont; Gry Kloumann Bekken; Stephen R Knight; Robin Kobbe; Chamira Kodippily; Malte Kohns Vasconcelos; Sabin Koirala; Mamoru Komatsu; ISARIC Collaborator Korten; Caroline Kosgei; Ars\u00e8ne Kpangon; Karolina Krawczyk; Vinothini Krishnan; Sudhir Krishnan; Oksana Kruglova; Deepali Kumar; Ganesh Kumar; Mukesh Kumar; Dinesh Kuriakose; Ethan Kurtzman; Demetrios Kutsogiannis; Galyna Kutsyna; Konstantinos Kyriakoulis; Marie Lachatre; Marie Lacoste; John G. Laffey; Marie Lagrange; Fabrice Laine; Olivier Lairez; Sanjay Lakhey; Antonio Lalueza; Marc Lambert; Fran\u00e7ois Lamontagne; Marie Langelot-Richard; Vincent Langlois; Eka Yudha Lantang; Marina Lanza; C\u00e9dric Laou\u00e9nan; Samira Laribi; Delphine Lariviere; St\u00e9phane Lasry; Sakshi Lath; Naveed Latif; Odile Launay; Didier Laureillard; Yoan Lavie-Badie; Andy Law; Teresa Lawrence; Cassie Lawrence; Minh Le; Cl\u00e9ment Le Bihan; Cyril Le Bris; Georges Le Falher; Lucie Le Fevre; Quentin Le Hingrat; Marion Le Mar\u00e9chal; Soizic Le Mestre; Gwena\u00ebl Le Moal; Vincent Le Moing; Herv\u00e9 Le Nagard; Paul Le Turnier; Ema Leal; Marta Leal Santos; Todd C. Lee; James Lee; Jennifer Lee; Heng Gee Lee; Biing Horng Lee; Yi Lin Lee; Su Hwan Lee; Gary Leeming; Laurent Lefebvre; B\u00e9n\u00e9dicte Lefebvre; Benjamin Lefevre; Sylvie LeGac; Jean-Daniel Lelievre; Fran\u00e7ois Lellouche; Adrien Lemaignen; V\u00e9ronique Lemee; Anthony Lemeur; Gretchen Lemmink; Ha Sha Lene; Jenny Lennon; Rafael Le\u00f3n; Marc Leone; Michela Leone; Quentin Lepiller; Fran\u00e7ois-Xavier Lescure; Olivier Lesens; Mathieu Lesouhaitier; Amy Lester-Grant; Bruno Levy; Yves Levy; Claire Levy-Marchal; Katarzyna Lewandowska; Erwan L'Her; Gianluigi Li Bassi; Janet Liang; Ali Liaquat; Geoffrey Liegeon; Wei Shen Lim; Kah Chuan Lim; Chantre Lima; Lim Lina; Bruno Lina; Andreas Lind; Maja Katherine Lingad; Guillaume Lingas; Sylvie Lion-Daolio; Keibun Liu; Marine Livrozet; Patricia Lizotte; Antonio Loforte; Navy Lolong; Leong Chee Loon; Diogo Lopes; Dalia Lopez-Colon; Jose W. Lopez-Revilla; Anthony L. Loschner; Paul Loubet; Bouchra Loufti; Guillame Louis; Silvia Lourenco; Lara Lovelace-Macon; Lee Lee Low; Marije Lowik; Jia Shyi Loy; Jean Christophe Lucet; Carlos Lumbreras Bermejo; Carlos M Luna; Olguta Lungu; Liem Luong; Nestor Luque; Dominique Luton; Nilar Lwin; Ruth Lyons; Olavi Maasikas; Oryane Mabiala; Mo\u00efse Machado; Gabriel Macheda; Hashmi Madiha; Guillermo Maestro de la Calle; Rafael Mahieu; Sophie Mahy; Ana Raquel Maia; Lars S. Maier; Myl\u00e8ne Maillet; Thomas Maitre; Maximilian Malfertheiner; Nadia Malik; Paddy Mallon; Fernando Maltez; Denis Malvy; Victoria Manda; Laurent Mandelbrot; Frank Manetta; Julie Mankikian; Edmund Manning; Aldric Manuel; Ceila Maria Sant\u2032Ana Malaque; Fl\u00e1vio Marino; Daniel Marino; Samuel Markowicz; Charbel Maroun Eid; Ana Marques; Catherine Marquis; Brian Marsh; Laura Marsh; Megan Marshal; John Marshall; Celina Turchi Martelli; Dori-Ann Martin; Emily Martin; Guillaume Martin-Blondel; Martin Martinot; Alejandro Martin-Quiros; Jo\u00e3o Martins; Ana Martins; Nuno Martins; Caroline Martins Rego; Gennaro Martucci; Olga Martynenko; Eva Miranda Marwali; Marsilla Marzukie; David Maslove; Sabina Mason; Sobia Masood; Basri Mat Nor; Moshe Matan; Meghena Mathew; Daniel Mathieu; Mathieu Mattei; Romans Matulevics; Laurence Maulin; Michael Maxwell; Javier Maynar; Thierry Mazzoni; Natalie Mc Evoy; Lisa Mc Sweeney; Colin McArthur; Colin McArthur; Anne McCarthy; Aine McCarthy; Colin McCloskey; Rachael McConnochie; Sherry McDermott; Sarah E. McDonald; Aine McElroy; Samuel McElwee; Victoria McEneany; Allison McGeer; Chris McKay; Johnny McKeown; Kenneth A. McLean; Paul McNally; Bairbre McNicholas; Elaine McPartlan; Edel Meaney; C\u00e9cile Mear-Passard; Maggie Mechlin; Maqsood Meher; Omar Mehkri; Ferruccio Mele; Luis Melo; Kashif Memon; Joao Joao Mendes; Ogechukwu Menkiti; Kusum Menon; Alexander J. Mentzer; Emmanuelle Mercier; No\u00e9mie Mercier; Antoine Merckx; Mayka Mergeay-Fabre; Blake Mergler; Ant\u00f3nio Mesquita; Roberta Meta; Osama Metwally; Agn\u00e8s Meybeck; Dan Meyer; Alison M Meynert; Vanina Meysonnier; Amina Meziane; Mehdi Mezidi; C\u00e9line Michelanglei; Isabelle Michelet; Efstathia Mihelis; Vladislav Mihnovit; Hugo Miranda-Maldonado; Nor Arisah Misnan; Tahira Jamal Mohamed; Nik Nur Eliza Mohamed; Asma Moin; David Molina; Elena Molinos; Brenda Molloy; Mary Mone; Agostinho Monteiro; Claudia Montes; Giorgia Montrucchio; Shona C. Moore; Sarah Moore; Lina Morales Cely; Lucia Moro; Catherine Motherway; Ana Motos; Hugo Mouquet; Clara Mouton Perrot; Julien Moyet; Caroline Mudara; Aisha Kalsoom Mufti; Ng Yong Muh; Dzawani Muhamad; Jimmy Mullaert; Fredrik M\u00fcller; Karl Erik M\u00fcller; Syed Muneeb; Nadeem Munir; Laveena Munshi; Aisling Murphy; Lorna Murphy; Aisling Murphy; Marl\u00e8ne Murris; Srinivas Murthy; Himed Musaab; Himasha Muvindi; Gugapriyaa Muyandy; Dimitra Melia Myrodia; Farah Nadia Mohd-Hanafiah; Dave Nagpal; Alex Nagrebetsky; Mangala Narasimhan; Nageswaran Narayanan; Rashid Nasim Khan; Alasdair Nazerali-Maitland; Nad\u00e8ge Neant; Holger Neb; Erni Nelwan; Raul Neto; Emily Neumann; Pauline Yeung Ng; Wing Yiu Ng; Anthony Nghi; Duc Nguyen; Orna Ni Choileain; Niamh Ni Leathlobhair; Prompak Nitayavardhana; Stephanie Nonas; Nurul Amani Mohd Noordin; Marion Noret; Nurul Faten Izzati Norharizam; Lisa Norman; Alessandra Notari; Mahdad Noursadeghi; Karolina Nowicka; Adam Nowinski; Saad Nseir; Jose I Nunez; Nurnaningsih Nurnaningsih; Dwi Utomo Nusantara; Elsa Nyamankolly; Fionnuala O Brien; Annmarie O Callaghan; Annmarie O'Callaghan; Giovanna Occhipinti; Derbrenn OConnor; Max O'Donnell; Tawnya Ogston; Takayuki Ogura; Tak-Hyuk Oh; Sophie O'Halloran; Katie O'Hearn; Shinichiro Ohshimo; Agnieszka Oldakowska; Jo\u00e3o Oliveira; Larissa Oliveira; Jee Yan Ong; Wilna Oosthuyzen; Anne Opavsky; Peter Openshaw; Saijad Orakzai; Claudia Milena Orozco-Chamorro; Jamel Ortoleva; Javier Osatnik; Linda O'Shea; Miriam O'Sullivan; Siti Zubaidah Othman; Nadia Ouamara; Rachida Ouissa; Eric Oziol; Ma\u00efder Pagadoy; Justine Pages; Mario Palacios; Amanda Palacios; Massimo Palmarini; Giovanna Panarello; Hem Paneru; Lai Hui Pang; Mauro Panigada; Nathalie Pansu; Aur\u00e9lie Papadopoulos; Rachael Parke; Melissa Parker; Briseida Parra; Taha Pasha; J\u00e9r\u00e9mie Pasquier; Bruno Pastene; Fabian Patauner; Drashti Patel; Mohan Dass Pathmanathan; Lu\u00eds Patr\u00e3o; Patricia Patricio; Juliette Patrier; Lisa Patterson; Rajyabardhan Pattnaik; Mical Paul; Christelle Paul; Jorge Paulos; William A. Paxton; Jean-Fran\u00e7ois Payen; Kalaiarasu Peariasamy; Giles J. Peek; Florent Peelman; Nathan Peiffer-Smadja; Vincent Peigne; Mare Pejkovska; Paolo Pelosi; Ithan D. Peltan; Rui Pereira; Daniel Perez; Luis Periel; Thomas Perpoint; Antonio Pesenti; Vincent Pestre; Lenka Petrou; Ventzislava Petrov-Sanchez; Frank Olav Pettersen; Gilles Peytavin; Scott Pharand; Michael Piagnerelli; Walter Picard; Olivier Picone; Maria de Piero; Carola Pierobon; Djura Piersma; Carlos Pimentel; Raquel Pinto; Catarina Pires; Isabelle Pironneau; Lionel Piroth; Ayodhia Pitaloka; Riinu Pius; Laurent Plantier; Hon Shen Png; Julien Poissy; Ryadh Pokeerbux; Maria Pokorska-Spiewak; Sergio Poli; Georgios Pollakis; Diane Ponscarme; Jolanta Popielska; Diego Bastos Porto; Andra-Maris Post; Douwe F. Postma; Pedro Povoa; Diana P\u00f3voas; Jeff Powis; Sofia Prapa; S\u00e9bastien Preau; Christian Prebensen; Jean-Charles Preiser; Anton Prinssen; Gamage Dona Dilanthi Priyadarshani; Lucia Proen\u00e7a; Sravya Pudota; Oriane Pu\u00e9chal; Bambang Pujo Semedi; Mathew Pulicken; Gregory Purcell; Luisa Quesada; Vilmaris Quinones-Cardona; V\u00edctor Quir\u00f3s Gonz\u00e1lez; Else Quist-Paulsen; Mohammed Quraishi; Maia Rabaa; Christian Rabaud; Ebenezer Rabindrarajan; Aldo Rafael; Marie Rafiq; Gabrielle Ragazzo; Mutia Rahardjani; Rozanah Abd Rahman; Ahmad Kashfi Haji Ab Rahman; Arsalan Rahutullah; Fernando Rainieri; Giri Shan Rajahram; Pratheema Ramachandran; Ahmad Afiq Ramli; Blandine Rammaert; Asim Rana; Rajavardhan Rangappa; Ritika Ranjan; Christophe Rapp; Aasiyah Rashan; Thalha Rashan; Ghulam Rasheed; Menaldi Rasmin; Indrek R\u00e4tsep; Cornelius Rau; Tharmini Ravi; Ali Raza; Andre Real; Stanislas Rebaudet; Sarah Redl; Brenda Reeve; Attaur Rehman; Liadain Reid; Liadain Reid; Dag Henrik Reikvam; Renato Reis; Jordi Rello; Jonathan Remppis; Martine Remy; Hongru Ren; Hanna Renk; Anne-Sophie Resseguier; Matthieu Revest; Oleksa Rewa; Luis Felipe Reyes; Tiago Reyes; Maria Ines Ribeiro; Antonia Ricchiuto; David Richardson; Denise Richardson; Laurent Richier; Siti Nurul Atikah Ahmad Ridzuan; Jordi Riera; Ana L Rios; Asgar Rishu; Patrick Rispal; Karine Risso; Maria Angelica Rivera Nu\u00f1ez; Nicholas Rizer; Chiara Robba; Andr\u00e9 Roberto; Stephanie Roberts; David L. Robertson; Olivier Robineau; Ferran Roche-Campo; Paola Rodari; Sim\u00e3o Rodeia; Bernhard Roessler; Pierre-Marie Roger; Emmanuel Roilides; Juliette Romaru; Roberto Roncon-Albuquerque Jr; M\u00e9lanie Roriz; Manuel Rosa-Calatrava; Michael Rose; Dorothea Rosenberger; Nurul Hidayah Mohammad Roslan; Andrea Rossanese; Matteo Rossetti; B\u00e9n\u00e9dicte Rossignol; Patrick Rossignol; Stella Rousset; Carine Roy; Beno\u00eet Roze; Desy Rusmawatiningtyas; Clark D. Russell; Maria Ryan; Maeve Ryan; Steffi Ryckaert; Aleksander Rygh Holten; Isabela Saba; Sairah Sadaf; Musharaf Sadat; Valla Sahraei; Maximilien Saint-Gilles; Pranya Sakiyalak; Nawal Salahuddin; Leonardo Salazar; Jodat Saleem; Gabriele Sales; St\u00e9phane Sallaberry; Charlotte Salmon Gandonniere; H\u00e9l\u00e8ne Salvator; Olivier Sanchez; Angel Sanchez-Miralles; Vanessa Sancho-Shimizu; Gyan Sandhu; Zulfiqar Sandhu; Pierre-Fran\u00e7ois Sandrine; Marlene Santos; Shirley Sarfo-Mensah; Bruno Sarmento Banheiro; Iam Claire E. Sarmiento; Benjamine Sarton; Ankana Satya; Sree Satyapriya; Rumaisah Satyawati; Egle Saviciute; Parthena Savvidou; Yen Tsen Saw; Justin Schaffer; Tjard Schermer; Arnaud Scherpereel; Marion Schneider; Stephan Schroll; Michael Schwameis; Gary Schwartz; Brendan Scicluna; Janet T. Scott; James Scott-Brown; Nicholas Sedillot; Tamara Seitz; Jaganathan Selvanayagam; Mageswari Selvarajoo; Caroline Semaille; Rasidah Bt Senian; Eric Senneville; Claudia Sepulveda; Filipa Sequeira; T\u00e2nia Sequeira; Ary Serpa Neto; Pablo Serrano Balazote; Ellen Shadowitz; Syamin Asyraf Shahidan; Mohammad Shamsah; Anuraj Shankar; Shaikh Sharjeel; Catherine A. Shaw; Victoria Shaw; Ashraf Sheharyar; Rohan Shetty; Dr Rajesh Mohan Shetty; Haixia Shi; Nisreen Shiban; Mohiuddin Shiekh; Nobuaki Shime; Hiroaki Shimizu; Keiki Shimizu; Sally Shrapnel; Pramesh Sundar Shrestha; Shubha Kalyan Shrestha; Hoi Ping Shum; Nassima Si Mohammed; Ng Yong Siang; Jeanne Sibiude; Atif Siddiqui; Piret Sillaots; Catarina Silva; Rog\u00e9rio Silva; Maria Joao Silva; Wai Ching Sin; Dario Sinatti; Punam Singh; Budha Charan Singh; Pompini Agustina Sitompul; Karisha Sivam; Vegard Skogen; Sue Smith; Benjamin Smood; Coilin Smyth; Michelle Smyth; Michelle Smyth; Morgane Snacken; Dominic So; Tze Vee Soh; Joshua Solomon; Tom Solomon; Agn\u00e8s Sommet; Rima Song; Tae Song; Jack Song Chia; Michael Sonntagbauer; Azlan Mat Soom; Alberto Sotto; Edouard Soum; Marta Sousa; Ana Chora Sousa; Maria Sousa Uva; Vicente Souza-Dantas; Alexandra Sperry; Elisabetta Spinuzza; B. P. Sanka Ruwan Sri Darshana; Shiranee Sriskandan; Sarah Stabler; Thomas Staudinger; Stephanie-Susanne Stecher; Trude Steinsvik; Ymkje Stienstra; Birgitte Stiksrud; Eva Stolz; Amy Stone; Adrian Streinu-Cercel; David Stuart; Ami Stuart; Decy Subekti; Gabriel Suen; Jacky Y. Suen; Asfia Sultana; Charlotte Summers; Dubravka Supic; Deepashankari Suppiah; Magdalena Surovcov\u00e1; Suwarti Suwarti; Andrey Svistunov; Sarah Syahrin; Konstantinos Syrigos; Jaques Sztajnbok; Konstanty Szuldrzynski; Shirin Tabrizi; Lysa Tagherset; Shahdattul Mawarni Taib; Ewa Talarek; Sara Taleb; Jelmer Talsma; Renaud Tamisier; Maria Lawrensia Tampubolon; Kim Keat Tan; Yan Chyi Tan; Taku Tanaka; Hiroyuki Tanaka; Hayato Taniguchi; Huda Taqdees; Arshad Taqi; Coralie Tardivon; Pierre Tattevin; M Azhari Taufik; Hassan Tawfik; Richard S. Tedder; Tze Yuan Tee; Jo\u00e3o Teixeira; Sofia Tejada; Marie-Capucine Tellier; Sze Kye Teoh; Vanessa Teotonio; Fran\u00e7ois T\u00e9oul\u00e9; Pleun Terpstra; Olivier Terrier; Nicolas Terzi; Hubert Tessier-Grenier; Adrian Tey; Alif Adlan Mohd Thabit; Anand Thakur; Zhang Duan Tham; Suvintheran Thangavelu; Vincent Thibault; Simon-Djamel Thiberville; Beno\u00eet Thill; Jananee Thirumanickam; Shaun Thompson; Emma C. Thomson; David Thomson; Surain Raaj Thanga Thurai; Ryan S. Thwaites; Paul Tierney; Vadim Tieroshyn; Peter S Timashev; Jean-Fran\u00e7ois Timsit; No\u00e9mie Tissot; Jordan Zhien Yang Toh; Maria Toki; Kristian Tonby; Sia Loong Tonnii; Margarida Torres; Antoni Torres; Rosario Maria Torres Santos-Olmo; Hernando Torres-Zevallos; Michael Towers; Tony Trapani; Th\u00e9o Treoux; C\u00e9cile Tromeur; Ioannis Trontzas; Tiffany Trouillon; Jeanne Truong; Christelle Tual; Sarah Tubiana; Helen Tuite; Jean-Marie Turmel; Lance C.W. Turtle; Anders Tveita; Pawel Twardowski; Makoto Uchiyama; PG Ishara Udayanga; Andrew Udy; Roman Ullrich; Alberto Uribe; Asad Usman; Timothy M Uyeki; Cristinava Vajdovics; Piero Valentini; Lu\u00eds Val-Flores; Am\u00e9lie Valran; Stijn Van de Velde; Marcel van den Berge; Machteld Van der Feltz; Job van der Palen; Paul van der Valk; Nicky Van Der Vekens; Peter Van der Voort; Sylvie Van Der Werf; Laura van Gulik; Jarne Van Hattem; Carolien van Netten; Frank van Someren Greve; Ilonka van Veen; Hugo Van Willigen; No\u00e9mie Vanel; Henk Vanoverschelde; Pooja Varghese; Michael Varrone; Shoban Raj Vasudayan; Charline Vauchy; Shaminee Veeran; Aur\u00e9lie Veislinger; Sebastian Vencken; Sara Ventura; Annelies Verbon; James Vickers; Jos\u00e9 Ernesto Vidal; C\u00e9sar Vieira; Deepak Vijayan; Joy Ann Villanueva; Judit Villar; Pierre-Marc Villeneuve; Andrea Villoldo; Gayatri Vishwanathan; Benoit Visseaux; Hannah Visser; Chiara Vitiello; Harald Vonkeman; Fanny Vuotto; Suhaila Abdul Wahab; Noor Hidayu Wahab; Nadirah Abdul Wahid; Marina Wainstein; Wan Fadzlina Wan Muhd Shukeri; Chih-Hsien Wang; Steve Webb; Katharina Weil; Tan Pei Wen; Sanne Wesselius; T. Eoin West; Murray Wham; Bryan Whelan; Nicole White; Paul Henri Wicky; Aur\u00e9lie Wiedemann; Surya Otto Wijaya; Keith Wille; Suzette Willems; Virginie Williams; Calvin Wong; Yew Sing Wong; Teck Fung Wong; Natalie Wright; Gan Ee Xian; Lim Saio Xian; Kuan Pei Xuan; Ioannis Xynogalas; Siti Rohani Binti Mohd Yakop; Masaki Yamazaki; Yazdan Yazdanpanah; Nicholas Yee Liang Hing; C\u00e9cile Yelnik; Chian Hui Yeoh; Stephanie Yerkovich; Toshiki Yokoyama; Hodane Yonis; Obada Yousif; Saptadi Yuliarto; Akram Zaaqoq; Marion Zabbe; Kai Zacharowski; Masliza Zahid; Maram Zahran; Nor Zaila Binti Zaidan; Maria Zambon; Miguel Zambrano; Alberto Zanella; Konrad Zawadka; Nurul Zaynah; Hiba Zayyad; Alexander Zoufaly; David Zucman; Mazankowski Heart Institute; The Western Australian COVID-19 Research Response.Ethics Committee approval was given by the WHO Ethics Review Committee . Institutional approval was additionally obtained by participating sites including the South Central\u2014Oxford C Research Ethics Committee in England (Ref. 13/SC/0149), the Scotland A Research Ethics Committee (Ref. 20/SS/0028) for the UK and the Human Research Ethics Committee at the University of the Witwatersrand in South Africa as part of a national surveillance programme (M160667), which collectively represent the majority of the data. Other institutional and national approvals are in place as per local requirements.dyad012_Supplementary_DataClick here for additional data file."} +{"text": "In the editorial section, Mohamed El Amine Youcef Ali et al. (550) list the difficulties of addressing social determinants associated with cancer outcomes. Bruce Gordon et al. (551) describe the extent to which unsafe water, sanitation and hygiene persist as a public health problem. Tatum Anderson (554 \u2013 555) reports on the use of new guidelines for the prevention and treatment of post-partum haemorrhage. Abla Mehio Sibai talks to Lynn Eaton (556 \u2013 557) about cross-disciplinary approaches to healthy ageing and the challenges posed by the collapsing public sector in Lebanon.Ajay Acharya et al. (571 \u2013 586) review verbal autopsy studies for attributable mortality.Agya Mahat et al. (595 \u2013 604) summarize recent reforms in six countries and discuss global implications. Stephen A Spencer et al. (558 \u2013 570) review observational studies of hospitalized adults.Christina Yek et al. (605 \u2013 616) summarize 18 years of national policies and surveillance data.C S Pramesh et al. (587 \u2013 594) report on a pilot initiative to pool procurement of antineoplastics."} +{"text": "Viroj Tangcharoensathien et al. (230) call for submissions to an upcoming theme issue on global health and equity. Lia Tadesse et al. (231) introduce a World Health Assembly item on the strengthening of emergency, critical and operative care. Fid Thomson (234\u2013235) reports on the need for a multisectoral response to cholera control in affected countries. Firdausi Qadri talks to Gary Humphreys (236\u2013237) about the need for new cholera vaccines and increased vaccine production.Daniel O\u2019Keefe et al. (262\u2013270) describe a pilot project initiated by nurses.Songchun Yang et al. (238\u2013247) evaluate performance of the risk prediction model in ten regions. Haishaerjiang Wushouer et al. (248\u2013261) track trends in inpatient use.Haoxiang Lin et al. (271\u2013280) conduct a randomized controlled trial.Shally Awasthi et al. (281\u2013289) validate a tool that evaluates risk of death.Veronica Zanichelli et al. (290\u2013296) present WHO\u2019s approach to preventing antimicrobial resistance."} +{"text": "In the study \u201cImplementing Symptom Management Follow-up Using an Electronic Patient-Reported Outcome Platform in Outpatients With Advanced Cancer: Longitudinal Single-Center Prospective Study,\u201d Tang et al suggest Tang et al reported"} +{"text": "In the editorial section, Manjulaa Narasimhan et al. (750) call for papers for a theme issue on sexual health and well-being across the life course. Alan D Lopez et al. (751) argue that health sector leadership is needed to improve civil registration and vital statistics systems.Gary Humphreys (754\u2013755) reports on the potential public health implications of a proposed global plastics treaty and interviews Rabih El Chammay (756\u2013757) about the importance of mental health in humanitarian emergencies.Kerolyn Shairsingh et al. (800\u2013807) introduce new countries in WHO\u2019s database.Ibrahem Abduallah Beshr et al. (808\u2013812) track vaccination campaigns.Lene Mikkelsen et al (758\u2013767) compare the performance of national civil registration and vital statistics systems.Tim Adair et al. present a global analysis of birth statistics (768\u2013776) and assess the policy utility of routine mortality statistics (777\u2013785). Brad Wong et al. (786\u2013799) review the evidence of economic benefits."} +{"text": "It is advanced as a more robust form of inference, with guaranteed frequentist properties.Peter Gr\u00fcnwald proposesChris Holmes & Stephen Walker connect Sonia Petrone also focet al. (2016) [Veronika Rockova & Lizhen Nie brings f. (2016) . A stron. (2016) and thatet al. [When seeking computational solutions to approximate an essential Bayesian entity, namely the Bayes factor (or the evidence), Arnaud Doucet et al. similarlAlicia Carriquiry & Kori Khan build a p-values, thus connecting with Peter Gr\u00fcnwald e-values. He also advocates considering alternative approaches for faster or more robust resolutions.Matthew Stephens contribuSylvia Fr\u00fcwirth-Schnatter broadenset al. 1984 [et al. (2016) [Sara Wade advocateal. 1984 ). She br. (2016) .Kerrie Mengersen & her co-authors also offRichard Nickl & co-authors study hiet al. [Peter M\u00fcller et al. open a nSumio Watanabe presentsBeatrice Franzolini & co-authors study an"} +{"text": "In the editorial section, Giorgio Cometto et al. (362) explain provisions of WHO\u2019s 2023 health workforce support and safeguards list. Katherine Pettus et al. (363) argue that a future pandemic treaty should include provisions for palliative care. Sameer Pujari et al. (364) advise that cautious optimism is warranted for artificial intelligence for global health.Tatum Anderson (367 \u2013 368) reports on projects to eliminate mercury in skin lightening products in Gabon, Jamaica and Sri Lanka. John Rex talks to Gary Humphreys (369 \u2013 370) about the challenges faced in developing new antibiotics and bringing them to market. Xingce Zhu et al. (381 \u2013 390) describe how artificial intelligence was used in a screening programme.Nduta Kamere et al. (403 \u2013 411) analyse supply chain factorsAdidja Amani et al. (431 \u2013 436) describe the roll-out of COVID-19 vaccination.Cathy Green et al. (371 \u2013 380) study the use of rectal artesunate.Barsha Gadapani Pathak et al. (391\u2013 402) review the evidence of effects on maternal and paternal health.Shatrunajay Shukla et al. (412 \u2013 417) discuss how to improve regulatory practicesFrank Pega et al. (418 \u2013 430) present a new global indicator for workers\u2019 health."} +{"text": "In the editorial section, Susanna Hausmann-Muela (494) describes the Botnar Foundation\u2019s view of WHO\u2019s urban health research agenda.Gary Humphreys (497 \u2013 498) reports on the incentives needed to support antibacterial innovation. Maria Asuncion Silvestre talks to Gary Humphreys (499 \u2013 500) about exclusive breastfeeding as part of neonatal care.Gerardo Priotto et al. (522 \u2013 528), (529 \u2013 534), (535 \u2013 540), (541 \u2013 545), (546 \u2013 548) present four target product profiles of diagnostic tests.Asri Maharani et al. (513 \u2013 521) study associations between coverage and access in a national family planning census.Scott JC Pallett et al. (501 \u2013 512) analyse data variability across surveillance platforms."} +{"text": "In the editorial section, Ahmad Reza Hosseinpoor et al. (298) present WHO's Health Inequality Data Repository. Ritu Sadana et al. (299) call for papers for an upcoming theme issue on building an economy for health. Rosamund F Lewis et al. (300) list the latest recommendations for countries responding to mpox. Tatum Anderson (303\u2013304) reports on the requirements for developing an effective tuberculosis vaccine. Vikram Patel talks to Gary Humphreys (305\u2013306) about developing evidence-based approaches to collaborative mental health service provision.Sarah C Charnaud et al. (326\u2013330) introduce a new feature in the Bulletin: WHO target product profiles.Wallace White et al. (331\u2013340) provide specifications for new readers of rapid diagnostic tests.Suman Rao et al. (341\u2013345) detail the aerosolized surfactants needed to treat neonatal respiratory distress.Matthias Helble et al. (355\u2013357) describes WHO\u2019s new coordinated scientific advice procedure and other services provided to product developers.Madhu Chhanda Mohanty et al. (346\u2013354) establish a surveillance network for patients with primary immunodeficiencies.Yafei Si et al. (307\u2013316) investigate how early-life factors are associated with intrinsic capacity.Jie Chen et al. (317\u2013325) track sales by pharmacies in the absence of prescriptions.Elizabeth K Dunford & Barry M Popkin (358\u2013360) describe global supply of, and demand for, ultraprocessed foods for children."} +{"text": "Sir,et al., with a great interest.[I read the interesting case report by Agarwal interest. There arinterest. The inteinterest.\u20136 The hointerest. A pre-diinterest.\u20136"} +{"text": "Mark Durand et al. (p. 1840), We regret any confusion this error may have caused."} +{"text": "We investigate the network model composed of subthalamic nucleus (STN) and globus pallidus (GP) developed by Nevado Holgado et al. who iden"} +{"text": "Rampin et al. recently published a report also describing social odor evoked activity in the olfactory tubercle:Rampin O, Bellier C, Maurin Y. Eur J Neurosci. 202 Jan;35(1):97-105.We regret not citing the publication by Rampin et al. Although the Payton et al. and Rampin et al. articles utilized different techniques and stimulus sets, together they begin to establish olfactory sensory coding by olfactory tubercle neurons."} +{"text": "In the editorial section, Marie-Ren\u00e9e B-Lajoie et al. (698) explain why Honduras needs to track urban epidemics of injuries and violent deaths more accurately. Iris Borowy (699) argues for WHO\u2019s involvement in drafting sustainable development goals.Petr T\u0159e\u0161\u0148\u00e1k (702\u2013703) reports on progress towards community-based mental health services in the Czech Republic. Fiona Fleck interviews Daniel Bausch (704\u2013705) on the current Ebola virus disease outbreak in western Africa.Mira Johri et al. (706\u2013715) model the effects of eliminating user fees for certain health services.Marta Miret et al. (716\u2013725) analyse findings from a study on ageing.Natacha Carragher et al. (726\u2013733) evaluate national alcohol control policies.Sujit\u00a0D Rathod et al. (734\u2013741) track all-cause mortality in communities that have gained access to treatment for HIV.Anja Takla et al. (742\u2013749) review surveillance data and assess underreporting.David\u00a0M Silvestri et al. (750\u2013759) describe medical and nursing students' reported intentions to work abroad or in rural areas.Eiko Saito et al. (760\u2013767) look for causes of catastrophic health spending.Rosemary Wyber et al. (768\u2013770) argue for global investment in programmes to prevent rheumatic heart disease.T.\u00a0cruzi.Yves Jackson et al. (771\u2013772) call for better recognition of, and treatment for, people infected with"} +{"text": "Recently, Lohr et al. have published a method that identifies sample annotation errors in gene expression data . SurpriCurrently, genome-wide data are frequently used in cancer research (Stock et al., 2015; SickingBesides its intensive use in cancer research (Micke et al., 2014; Schmidt"} +{"text": "In the editorial section, Alistair Woodward (774) reviews progress on climate change and health. Pat Oungpasuk (775) argues that all countries in the Association of South-East Asian Nations should adopt universal health coverage to ensure future economic growth. In a focus on WHO guidelines, Jim Mann tells Fiona Fleck (780\u2013781) how WHO surveyed the evidence for new recommendations on sugar intake. Priya Shetty (778\u2013779) reports on recent efforts to make WHO recommendations on HIV treatment more responsive to the realities facing programme managers.Terence\u00a0T Lao et al. (782\u2013789) track hepatitis\u00a0B prevalence in previously-vaccinated pregnant women in Hong Kong SAR.Takashi Kameda et al. (790\u2013797) study asbestos-related diseases in the context of national policies on its use.Michael Abouyannis et al. (798\u2013806) find a low prevalence of multidrug-resistant tuberculosis. Krycia Cowling et al. (817\u2013825) make the case for better tuberculosis estimates.United Kingdom of Great Britain & Northern IrelandReaching far-flung islandsEsther\u00a0L Hamblion et al. (836\u2013843) study compliance by overseas territories with the International Health Regulations.Augustine Bilve et al. (844\u2013848) describe the set-up of an early response network.Rasika Rampatige et al. (807\u2013816) present a method for reviewing the accuracy of cause-of-death statistics.Sarah\u00a0L White et al. (826\u2013835) discuss globalization of organ transplants."} +{"text": "In the editorial section, Gyambo Sithey et al. (514) discuss Bhutan\u2019s continued use of gross national happiness as a measure of development.In the news section, Andrei Shukshin reports on WHO\u2019s efforts to boost health worker capacities in Ukraine (517\u2013518). Fiona Fleck interviews Pascal Michel on the use of space technologies for public health (519\u2013520).Pablo F Belaunzar\u00e1n-Zamudio et al. document infrequent monitoring of patients receiving antiretrovirals. (529\u2013539)Jacques DA Ndawinz et al. propose measuring time-to-treatment for HIV. (521\u2013528)Helen Jack et al. highlight the difficulties of providing mental health care. (587\u2013588)Yuko Kumagi et al. adjust national estimates of major foodborne diseases.(540\u2013549)water quality in refugee camps. (550\u2013558)Syed Imran Ali et al. studyAnthony L Bui et al. review all available national health expenditure data. (566\u2013576)Ziad Obermeyer et al. analyse the provision of emergency care in low-resource settings.(577\u2013586)Frank Pega et al. describe cash transfers designed to mitigate the health effects of climate change. (559\u2013565)"} +{"text": "In the editorial section, Vasee S Moorthy et al. (234) present operating principles for health-data platforms. Viroj Tangcharoensathien et al. (235) introduce an upcoming theme issue on vulnerable populations. Sophie Delaunay et al. (236) discuss improved access to information during public health emergencies.Andr\u00e9ia Azevedo Soares reports on the effects of Mexico\u2019s soda tax (239\u2013240). Fiona Fleck interviews KM Venkat Narayan about the increased prevalence of type 2 diabetes (241\u2013242). Eva A Rehfuess et al. (297\u2013305) describe a priority-setting approach.Christine Goeppel et al. (276\u2013285) document the extent of health coverage for adults with chronic illness.Rakhi Dandona et al. (286\u2013296) review national health surveys.Leveraging drug vendors Jenny Liu et al. (267\u2013275) count drug shops, assess stocks and interview vendors.Almamy Malick Kant\u00e9 et al. (258\u2013266) study trends in socioeconomic disparities.Manjulaa Narasimhan et al. (243\u2013249) survey women about their reproductive health rights.Mark Goodchild et al. (250\u2013257) model the impact of raising tobacco taxes.Abdul Ghaffar et al. (306\u2013308) discuss a relative lack of health policy research."} +{"text": "After publication of the original article , it cameFondo de Investigaciones Sanitarias\u2014Health Research Fund, Grant No. PI13/00118, Instituto de Salud Carlos III), co-financed by the European Union through the Fondo Europeo de Desarrollo Regional , and by the Grupo de Excelencia Investigadora URJC-Banco Santander No. 30VCPIGI03: Investigaci\u00f3n traslacional en el proceso de salud\u2014enfermedad (ITPSE).\u201d\u201cThis study was funded by the FIS ("} +{"text": "In the editorial section, Ahmad Reza Hosseinpoor et al. (591) stress the importance of monitoring health inequality in the post-2015 agenda. James Campbell et al. (590) put the cost\u2013effectiveness study done by Barbara McPake et al. (631\u2013640) into the global context of using community-based practitioners to expand access to health care. In the news section, Apiradee Treerutkuarkul and Karl Gruber report on efforts to improve oral health in low- and middle-income countries (594\u2013595). Sheila Tlou tells Fiona Fleck how HIV prevention campaigns have evolved in Botswana since the 1980s (596\u2013597).Adolfo Rubinstein et al. (614\u2013622) estimate the health and economic impacts of a national policy to eliminate trans fats. Emmanuel Fajardo et al. (623\u2013630) examine invalid test results generated by a CD4+ analyser from sites in nine countries. Lei Gao et al. (661\u2013662) consider cultural influences on informed consent and disclosure practices. Olivier J Wouters & Panos G Kanavos (606\u2013613) consider potential policy solutions for high costs in the pharmaceutical sector. Barbara McPake et al. (631\u2013640) analyse the cost\u2013effectiveness of community-based practitioner programmes. Yuriko Suzuki et al. (598\u2013605) investigate the perception of radiation risks and psychological distress among evacuees of the Fukushima Daiichi nuclear power plant accident.Samath D Dharmaratne et al. (641\u2013648) analyse trends over 75 years in road traffic crashes, injuries and deaths. Thidar Pyone et al. (649\u2013660) review the tools available for rapid assessments of maternal and child health. David Bishai et al. (663\u2013664) explain why communities affected by a public health problem should describe and address it."} +{"text": "Global strategy for women\u2019s, children\u2019s and adolescents\u2019 health (2016\u20132030). This month\u2019s issue has a special theme which focuses on the implementation of the In the editorial section, Flavia Bustreo et al. (310) highlight the ongoing need to analyse what works to achieve the objectives of this strategy: survive, thrive and transform. Ian Askew et al. (311) remind readers why sexual and reproductive health and rights need to be addressed in the response to health emergencies. Zanele Mabaso et al. (312) ascribe the inclusion of adolescent health outcomes in the global strategy to young people\u2019s participation, and Nila Moeloek & Kesetebirhan Admasu (313) explain how ministers of health can help ensure positive change in their countries. Bethlehem Kiros (316\u2013317) reports on efforts to improve health outcomes for women and children living in rural Ethiopia. Sophie Cousins (318\u2013319) describes how chances of survival during pregnancy and childbirth have greatly improved in Nepal. Joannie Bewa tells Fiona Fleck (320\u2013321) how campaigns for sex education and free contraception are changing reproductive health prospects for young people in Benin.Syed Masud Ahmed et al. (351\u2013361) analyse national success factors.Preeti H Negandhi et al. (370\u2013375) describe a maternal infant death review system.Jennifer A Newberry et al. (388\u2013392) introduce an emergency response line for women confronting violence.Obed Dolo et al. (383\u2013387) describe a programme to train midwives to do surgical procedures.Teresa Murgu\u00eda-Peniche et al. (322\u2013330) study trends and distributions of stillbirths.Margaret C Hogan et al. (362\u2013369) reclassify causes of maternal deaths.Jeanne Chai et al. (331\u2013339) study the association between child growth and violence in the home.Britt McKinnon et al. (340\u2013350) estimate the prevalence of suicidal ideation in 32 low- and middle-income countries.Cicely Marston et al. (376\u2013382) explain why community participation is crucial.Laura Frost et al. (393\u2013395) explain why many voices are needed to understand health and development issues. Shyama Kuruvilla et al. (398\u2013400) outline a roadmap for implementation.Claudia Garc\u00eda-Moreno & Avni Amin (396\u2013397) argue that development goals will only be met if violence elimination targets are reached.Pascal Bijleveld et al. (401\u2013404) list the challenges faced by countries."} +{"text": "Antimicrobial resistance is an increasing concern in ICUs worldwide. Infection with an antibiotic resistant (ABR) strain of an organism is associated with greater mortality than infection with the non-resistant strain, but there are few data assessing whether being admitted to an intensive care unit (ICU) with high levels of antimicrobial resistance is associated with a worse outcome than being admitted to an ICU with low rates of resistance. The aim of this study was, therefore, to compare the characteristics of infections and antibiotic treatments and patient outcomes in patients admitted to ICUs in countries considered as having high levels of antibiotic resistance and those admitted to ICUs in countries considered as having low levels of antibiotic resistance.Data from the large, international EPIC II one-day point prevalence study on infections in patients hospitalized in ICUs were used. For the current study, we compared the data obtained from patients from two groups of countries: countries with reported MRSA rates of\u2009\u2265\u200925% and countries with MRSA rates of\u2009<\u20095% .On the study day, 1187/2204 (53.9%) patients in the HighABR ICUs were infected and 255/558 (45.7%) in the LowABR ICUs (P\u2009<\u20090.01). Patients in the HighABR ICUs were more severely ill than those in the LowABR ICUs, as reflected by a higher SAPS II score and had longer median ICU (12 days vs 5 days) and hospital (24 days vs 16 days) lengths of stay. They also had higher crude ICU (20.0% vs 15.4%) and hospital (27.0% vs 21.5%) mortality rates (both P\u2009<\u20090.05). However, after multivariable adjustment and matched pair analysis there were no differences in ICU or hospital mortality rates between High or LowABR ICU patients overall or among those with infections.Being hospitalized in an ICU in a region with high levels of antimicrobial resistance is not associated per se with a worse outcome.The online version of this article (doi:10.1186/1471-2334-14-513) contains supplementary material, which is available to authorized users. The prevalence of infection is high among patients admitted to ICUs and is one of the main causes of ICU mortality , 2. In 1Antimicrobial resistance is an increasing concern in ICUs worldwide. Rates of resistance vary considerably across different countries and regions; for example, ICUs in southern Europe generally have higher rates of resistance than countries in northern Europe and Scandinavia . SeveralThe worldwide EPIC II 1-day point-prevalence study of infection and demographics of critically ill patients was performed on 8 May 2007 . MicrobiStaphylococcus aureus (MRSA) reported in the European Antimicrobial Resistance Surveillance System (EARSS) 2007 Annual Report [For the purposes of this analysis, we selected two groups of countries based on the country rate of methicillin-resistant l Report : countriThe EPIC II study was approved by the ethics committee of Erasme Hospital, Belgium, the coordinating center. Local ethical committee approval at each participating center (see Appendix for list of participating centers) was expedited or waived because of the observational nature of the study.\u03c72 test, or Fisher\u2019s exact test as appropriate. Logistic regression analysis was used to determine the mortality risk associated with admission to an ICU in a country with high levels of antimicrobial resistance or one with low levels of antimicrobial resistance. To remove any bias of confounding variables for the association between the areas of ICU admission and mortality, a propensity score was estimated for each of the ICU areas, with the following variables considered as factors: type of admission, source of admission, comorbidities, age, sex, mechanical ventilation, haemofiltration or haemodialysis, infection, SAPS II score and type of microorganism. After checking that balance on all covariates that were used in the propensity model had been achieved, odds ratios with mortality as the dependent variable were estimated by logistic regression using two strategies: by matching and by introducing the propensity score in the model [Statistical analyses were performed using IBM SPSS Statistics 20 for Windows . The Kolmogorov-Smirnov test was used, and histograms and Q-Q plots were examined to verify if there were significant deviations from the normality assumption of continuous variables. Difference testing between groups was performed using Mann-Whitney test, he model , 13. Rephe model ) and geoIn the EPIC II database, there were 2204 patients included from ICUs in countries with high levels of antimicrobial resistance (HighABR) according to the EARSS 2007 Annual Report and 558 Pseudomonas aeruginosa or Actinetobacter were significantly more prevalent among HighABR ICU patients (Table\u00a0S. aureus (MSSA), enterococci and anaerobes were more common in LowABR ICU patients.More patients in the HighABR ICUs than in the LowABR ICUs were infected on the study day . The respiratory system was the most common site of infection in all patients . When considering groups of antimicrobials, carbapenems, aminoglycosides, quinolones and glycopeptides were more commonly used in patients in HighABR ICUs than in LowABR ICUs, whereas cephalosporins were used more frequently as treatment in LowABR ICUs Table\u00a0. For indUse of prophylactic antimicrobials was significantly less common in HighABR ICU patients than in LowABR ICU patients . Among the agents most commonly used were aminoglycosides (tobramycin), antifungals (amphotericin B) and cephalosporins and hospital (27.0% vs 21.5%) mortality rates were higher in patients admitted to HighABR than in those admitted to LowABR ICUs (both P\u2009<\u20090.05); however, these differences were not present after adjustment in multivariable analysis or the matched pair analysis ; N\u00e6stved Hospital (B Fogh); Rigshospitalet (K Espersen); Sygehus Fyn (K Jacobsen); Vejle Sygehus (P Berezowicz); Finland: Helsinki University Central Hospital (V Harjola); Greece: Ahepa University Hospital (E Sofianos); Athens University Medical School (A Armaganidis); Evangelismos Hospital (C Routsi); G.Papanikolaou (M Bitzani); General Hospital of Rethymno ; Henry Dunant Hospital ; Hippokrateion Hospital Thessaloniki (E Mouloudi); Kat General Hospital (E Ioannidou); Kat Hospital (P Myrianthefs); Kat Hospital, Athens (D Koulenti); Konstantopoulio General Hospital (I Karampela); Lamia General Hospital (G Kyriazopoulos); Red Cross Hospital of Athens (K Mandragos); Thriassio Hospital of Eleusis (P Clouva-molyvdas); University Hospital of Ioannina (A Moraiti); University Hospital of Alexandroupolis (I Pneumatikos); University Hospital of Rion, Patras (K Filos); University Hospital of Thessaly (Larissa) (E Zakynthinos); University of Athens, Medical Shcool (A Kotanidou); Xanthi General Hospital ; Israel: Hadassah Medical Center (C Sprung); Haemek Medical Center (A Lev); Kaplan Medical Center (E Kishinevsky); Rabin Medical Center (J Cohen); Soroka Medical Center (S Sofer) Italy: A.O Niguarda (S Vesconi); A.O. Ospedale Di Circolo Di Busto Arsizio (S Greco); A.O. Treviglio-Caravaggio (M Borelli); Anestesia E Rianimazione 2 Prof.De Gaudio (P Cecilia); Arnas Ospedale Civico (M Sapuppo); ASL 10 (A Lazzero); ASL 10 Florence Hospital San Giovanni Di Dio (V Mangani); Azienda Ospedaliera Desenzano (N Petrucci); Azienda Ospedaliera Di Melegnano (M Minerva); Azienda Ospedaliera G. Rummo (E De blasio); Azienda Ospedaliera Polo Universitario San Paolo (S Marzorati); Azienda Ospedaliera Santa Maria Alle Scotte (R Rosi); Azienda Ospedaliera Universitaria P.Giaccone Policlinico (A Giarratano); Azienda Ospedaliera-Universitaria Udine (O Margarit); Azienda Ospedaliero -Universitaria (A Guberti); Azienda Ospedaliero-Universitaria S.M.Misericordia (S Scolz); Clinica San Gaudenzio (E Stelian); Fondazione IRCCS Policlinico San Matteo (V Emmi); Fondazione Ospedale Maggiore Policlinico, Mangiagalli Regina Elena (M Caspani); Fondazione Poliambulanza (A Rosano); H San Gerardo (C Abbruzzese); Hospital Panico Tricase (S Colonna); Humanitas Gavazzeni (R Ceriani); II Faculty of Medicne I University of Rome- Osp. S.Andrea (R De Blasi); S. Salvatore Hospital (L Panella); IRCCS Casa Sollievo Della Sofferenza (F Borrelli); Istituto Nazionale Tumori Regina Elena (P Lorella); KH Brixen (H Ruatti); Osepdali Riuniti Di Ancona (C Munch); Ospedale \"Ca Foncello\" - Treviso (C Sorbara); Ospedale \"Santa Croce\" - ASL 8 (G Fiore); Ospedale Bufalini-Cesena (A Chieregato); Ospedale Di Circolo E Fondazione Macchi (V Conti); Ospedale Di Massa (A Guadagnucci); Azienda USL Piacenza (M Pizzamiglio); Ospedale Ferrarotto (M Locicero); Ospedale Maggiore Ausl Bologna (I Marri); Ospedale Maggiore Policlinico Milano (A Sicignano); Ospedale Maggiore Policlinico, Mangiagalli E Regina Elena, IRCCS Milano (V Conte); Ospedale Mugello Azienda Sanitaria Firenze (R Oggioni); Ospedale Niguarda Ca Granda, Milano (A De Gasperi); IRCCS Centro di Riferimento Oncologico della Basilicata (P De Negri); Ospedale Provinciale Pistoia (G Santagostino); Ospedale S. Gerardo ; Ospedale San Raffaele (G Marino); Ospedele Vittorio Emanuele (G Castiglione); P.O. San Severo Asl Fg (D Sforza); S. Camillo Hospitql (N Giuseppe); San Martino Hospital (M Bassetti); Seconda Universit\u00e0 Degli Studi Di Napoli (F Ferraro); Sesto San Giovanni Hospital (S Clementi); Teaching Hospital Careggi (D Alessandro); Terapia Intensiva - Aso S. Giovanni Battista Di Torino - Ospedale Molinette ; Universit\u00e0 Cattolica (M Antonelli); Universita' Cattolica Del S. Cuore (L Martinelli); University-Hospital Careggi, Florence, (L Gianesello); University Hospital Policlinico Di Catania (A Gullo); University of Rome \"La Sapienza\" (A Morelli); UTI Trapianti (G Biancofiore); University of Udine (G Della Rocca) Malta: St Luke\u2019s Hospital (M Borg); Netherlands: Academic Medical Center (A De Pont); Amphia Hospital (P Rosseel); Antoni Van Leeuwenhoek Ziekenhuis (J Ten Cate); Beatrix Zienhuis Rivas Zorggroep (G Van Berkel); Canisius Wilhelmina Ziekenhuis (S Corsten); Erasmus Mc University Medical Center (J Bakker); Hagaziekenhuis (J Vogelaar); Hofpoort Hospital Woerden (H Blom); Isala Clinics (H Kieft); Medical Center Leeuwarden (M Kuiper); Medisch Spectrum Twente (A Gille); Radboud University Nijmegen Medical Centre (P Pickkers); Rode Kruis Ziekenhuis (J Vet); Slingeland Ziekenhuis (J Ammann); Spaarneziekenhuis (S Den Boer); St. Antonius Ziekenhuis (R Wesselink); St.Elisabeth Hospital (B Speelberg); Twenteborg Hospital Almelo (C Pham); University Medical Center, Groningen (M Rodgers); University Hospital Maastricht (D Bergmans); Vu University Medical Center (J Groeneveld); Norway: Aker University Hospital (R Loevstad); St Olavs University Hospital (P Klepstad); Sykehuset Asker Og B\u00e6rum Hf (P Erno); Sykehuset I Vestfold Hf, Toensberg (A Junker); Portugal: Centro Hospitalar Alto Ave (A B\u00e1rtolo); Centro Hospitalar Cova Da Beira (M Castelo-Branco Sousa); Centro Hospitalar Tr\u00e1s os Montes e Alto Douro (F Esteves); CHLO-Hospital S Francisco Xavier (A Martins); H S Jo\u00e3o (T Oliveira); Hospital CUF Infante Santo (P Ponce); Hospital Curry Cabral (L Mour\u00e3o); Hospital da Luz (C Febra); Hospital de Egas Moniz (E Carmo); Hospital de S. Jos\u00e9 (V Lopes); Hospital de S\u00e3o Francisco Xavier (P P\u00f3voa); Hospital de S\u00e3o Jos\u00e9 (A Rezende); Hospital Divino Espirito Santo (H Costa); Hospital do Litoral Alentejano (P Moreira); Hospital Dr. Jos\u00e9 Maria Grande, Portalegre (F P\u00e1dua); Hospital Fernando Fonseca (A Leite); Hospital Garcia de Orta (E Almeida); Hospital Geral de Santo Ant\u00f3nio (M Alves); Hospital de Pulido Valente ; Hospital de S. Jo\u00e3o ; Hospital de S\u00e3o Bernardo (R Ribeiro); Hospital de S\u00e3o Sebasti\u00e3o, EPE (P Amaro); Hospital Geral de Sto Ant\u00e1nio (A Carneiro); Hospital de St. Ant\u00f3nio dos Capuchos ; Instituto Portugu\u00eas de Oncologia de Lisboa (M Bouw); Hospital de St Maria (C Fran\u00e7a); Spain: Althaia (O Rubio); Bellvitge University Hospital (R Ma\u00f1ez); Centro Medico Delfos (M Burgue\u00f1o Campi\u00f1ez); Clinica Moncloa (M Alvarez); Clinica Rotger (R Jorda); Clinica Santa Elena (E Naveira-Abeig\u00f3n); Clinica Universitaria de Navarra (P Monedero); Complejo Hospitalario de Pontevedra (E Alemparte-Pardavila); Fundacion Hospital Alcorcon ; Fundacion Jimenez Diaz ; H Vall Hebron ; H.U. Virgen de Las Nieves- H. Traumatolog\u00eda Y Rehabilitaci\u00f3n (F Guerrero); Hospital \"Virgen de La Concha\" - Zamora ; Hospital Arnau de Vilanova (M Arribas); Hospital Can Misses (E Bustamante Munguira); Hospital Casa de Salud (J Ruiz); Hospital Central de Asturias (L Iglesias); Hospital Clinic ; Hospital Clinic de Barcelona ; Hospital Clinico San Carlos ; Hospital Clinico San Carlos ; Hospital Clinico Universitario (G Aguilar); Hospital Clinico Universitario de Santiago (F Martinon-Torres); Hospital Comarcal Vinaros (C Lorente); Hospital de Navarra (J Insausti); Hospital de Antequera (R Vegas Pinto); Hospital de Basurto (I Santos); Hospital de Fuenlabrada (A Escriba); Hospital de Galdakao (P Olaechea); Hospital de La Merced (E Mu\u00f1oz); Hospital de Manacor ; Hospital de Mostoles ; Hospital de Sagunto (V Lopez Camps); Hospital de Tortosa Verge de La Cinta (F Esteban-Reboll); Hospital del Sas de Jerez (A Estella); Hospital Donostia (L Bocero); Hospital Dr Peset (A Iba\u00f1ez); Hospital G. Yag\u00fce (L Pueyo); Hospital General (L Mar\u00eda Jes\u00fas); Hospital General de Asturias (L Iglesias); Hospital General de Ciudad Real (J Silva); Hospital General de Granollers (P Garro); Hospital General de La Palma (L Ramos-g\u00f3mez); Hospital General de L'Hospitalet (A Rovira); Hospital General de Vic (M Martin Delgado); Hospital General Salud \"Obispo Polanco\" (J Monton Dito); Hospital General Universitario de Albacete (F Garcia); Hospital General Universitario de Alicante (J Navarro); Hospital General Universitario de Elche (J Latour-Perez); Hospital General Universitario de Guadalajara (A Albaya); Hospital General Universitario Gregorio Mara\u00f1on (A Bustinza); Hospital Gran Canaria \"Dr Negr\u00edn\" (J Sole-viol\u00e1n); Hospital Marques de Valdecilla (P Ugarte Pe\u00f1a); Hospital Maz (I Yuste); Hospital Parque San Antonio (J De Rojas Rom\u00e1n); Hospital Sabadell ; Hospital Sant Joan de D\u00e9u (E Esteban); Hospital Sant Pau (E Quintana Tort-Martorell); Hospital Santa Mar\u00eda del Rosell (M Moreno); Hospital Santa Maria Madre-Complejo Hospitalario de Ourense (V L\u00f3pez Ciudad); Hospital Santiago Apostol (A Manzano Ramirez); Hospital Sevilla-Aljarafe (J S\u00e1nchez-Olmedo); Hospital Son Ll\u00e0tzer (M Borges); Hospital Terrassa (J Amador Amerigo); Hospital Torrecardenas (F Guerrero Gomez); Hospital Universitaio 12 de Octubre (J Montejo Gonz\u00e1lez); Hospital Universitari de Girona Doctor Josep Trueta (J Sirvent); Hospital Universitari Germans Trias I Pujol ; Hospital Universitario Arnau de Vilanova (F Barcenilla-Gaite); Hospital Universitario de Canarias (N Serrano); Hospital Universitario de Getafe (E Cerd\u00e1); Hospital Universitario de Valme (A Lesmes Serrano); Hospital Universitario Doce de Octubre (C Garcia-Fuentes); Hospital Universitario Infanta Crsitina (J Macias Pingarr\u00f3n); Hospital Universitario N\u00aasra de Candelaria (E Espinosa); Hospital Universitario Principe de Asturias (M Sanchez Garcia); Hospital Universitario Reina Sof\u00eda, Murcia (F Felices); Hospital Universitario Virgen de La Victoria (M de la Torre-Prados); Hospital Univesitario Puerto Real (H Maria Jesus); Hospital Valle del Nalon (V Luis); Hospital Virgen Arrixaca (R Jara); Hospital Xanit Internacional (M Briones Lopez); Hospital Xeral Cies (P Posada); Hu La Paz ; Hu La Paz ; Joan Xxiii University Hospital (J Rello); Morales Meseguer (B Gil); Puerta del Mar Universitary Hospital (R Sierra); Rio Hortega University Hospital (J Rico-Feijoo); San Pedro de Alcantara Hospital (C Corcobado M\u00e1rquez); Servicio Navarro de Salud.Hospital Virgen del Camino (J Izura); Uci H. U. Salamanca (J Gonz\u00e1lez); Universitary Hospital Dr. Peset (J Soto Ib\u00e1\u00f1ez); Sweden: Anestesikliniken (P Petersen); Centralsjukhuset Karlstad (L Johansson); University Hospital, Link\u00f6ping ; G\u00f6teborg (I Lindstr\u00f6m); K\u00e4rnsjukhuset Sk\u00f6vde (A Paulsson); Karolinska University Hospital Huddinge ; Karolinska University Hospital, Solna (J Petersson); Lund University Hospital (H Friberg); Malmoe University Hospital (V Einar); Op/Iva Kliniken (F Hammarskj\u00f6ld); Ostersund Hospital (M Schindele); Ostra Hospital, G\u00f6teborg (S Arvidsson); Sahlgrenska University Hospital (J Sellgren); S\u00f6der Hospital (S\u00f6dersjukhuset) (J Hulting); Sodersjukhuset (J H\u00e4ggqvist); Sollefte\u00e5 Hospital (J Rudenstam); Sunderby Hospital (D Lind); The Queen Silvia Children\u2019s Hospital (E Kokinsky); Thoracic Intensive Care, Karolinska Hospital ; Ume\u00e5 University Hospital (S Jacobson); University Hospital (H Stiernstrom); University Hospital of \u00d6rebro (A Nydahl); Turkey: Acibadem Kadikoy Hospital ; Acibadem Bakirkoy Hospital (C Ates); Acibadem Bursa Hospital (A Kahveci); Acibadem Kozyatagi Hospital (H Fistikci); Ankara Univercity (A Kaya); Ankara University Medical Faculty, Ibni Sina Hospital (E Ozgencil); Ataturk University Medical Faculty (M Kizilkaya); Dicle University Medical School (M Bosnak); Dokuz Eylul University (H Bodur); Dokuz Eylul University School of Medicine (M Akan); Erciyes University Medical Faculty (M Guven); Gazi University School of Medicine (M Turkoglu); Hacettepe University Hospital (A Topeli); Inonu University Medical Faculty ; Istanbul Faculty of Medicine (N Uzel); Istanbul Medical Faculty ; Istanbul University Cerrahpasa Medical School (O Demirkiran); Izmir Ataturk Training and Resarch Hospital (T Adanir); Memorial Hospital (K Dogruer); Okmeydani Teaching & Research Hospital (A Turkmen); Okmeydani Teaching & Research Hospital (H Guven); Ondokuz Mayis University, Medical Faculty (F Ulger); Selcuk University Meram Faculty of Medicine (S Kocak).Additional file 1: Sites of infection.(PDF 93 KB)Additional file 2: Use of antimicrobials as prophylaxis in all patients.(PDF 100 KB)"} +{"text": "Dear Editor,Catharanthus roseus . Treatmal., 1958; Johnsonal., 1958; Svobodaal., 1958) as wellal., 1971). One efPsychotria ipecacuanha Stokes (Rubiaceae) which is also known as Cephaelis ipecacuanha A. Rich (ipecac) where it is the principal alkaloid -2-{methyl}-3-ethyl-9,10-dimethoxy-2,3,4,6,7,11b-hexahydro-1H-pyrido isoquinoline Figure 1, is an ial., 1984). It is al., 1984) and it al., 1984). The bial., 1984).in vitro experimental studies requiring inhibition of protein biosynthesis but sinal., 2012). The daal., 2012) has shoal., 2012; Kim et al., 2012; Mayank al., 2012; Han et al., 2012; Myhren al., 2012; Foremanal., 2012; Akinboyal., 2012; Larrsonal., 2012; Pan et al., 2012; Kong etal., 2012). It is"} +{"text": "Melatonin is a neuroendocrine hormone synthesized primarily by the pineal gland. Numerous studies have suggested that melatonin plays an important role in various cardiovascular diseases. In this article, recent progress regarding melatonin's effects on cardiovascular diseases is reviewed.In the past year, studies have focused on the mechanism of protection of melatonin on cardiovascular diseases, including myocardial ischemia-reperfusion injury, myocardial hypoxia-reoxygenation injury, pulmonary hypertension, hypertension, atherosclerosis, valvular heart diseases, and other cardiovascular diseases.Studies have demonstrated that melatonin has significant effects on ischemia-reperfusion injury, myocardial chronic intermittent hypoxia injury, pulmonary hypertension, hypertension, valvular heart diseases, vascular diseases, and lipid metabolism. As an inexpensive and well tolerated drug, melatonin may be a new therapeutic option for cardiovascular disease. Melatonin (N-acetyl-5-methoxytryptamine) is a neuroendocrine hormone, which is synthesized primarily by the pineal gland . The synet al.[et al.[Melatonin confers profound protective effects against ischemia-reperfusion injury in various organs, including the heart ,8, liveret al. reportedet al.. The lacet al.. Anotheret al.\u25aa. Similaet al.. Its proet al.. Yu et al.[et al. studied l.[et al.. In an il.[et al.. Anotherl.[et al.. Melatonl.[et al.. In a ral.[et al.. In a mol.[et al.. In anotl.[et al.\u25aa, treatml.[et al.. Melatonl.[et al.\u25aa. A recel.[et al.\u25aa. In isol.[et al.\u25aa.et al.[et al.[et al.[et al.[Obstructive sleep apnea is associated with CIH and increases myocardial injury contributing to ischemic heart disease . Yeung eet al.\u25aa reportel.[et al. reportedl.[et al. demonstrl.[et al.. Importal.[et al. found thl.[et al..et al.[et al.[et al.[Pulmonary hypertension is a disease characterized by elevated pulmonary arterial pressure, which leads to right ventricular hypertrophy and failure . Maarmanet al. reportedl.[et al. found thl.[et al.\u25aa reporteet al.[et al.[Several studies have demonstrated that melatonin has an antihypertensive effect ,43. Simket al. found thl.[et al. also repl.[et al.. It was l.[et al.. Additiol.[et al..et al.[et al.[et al.[Recent studies have shown that melatonin is associated with atherosclerosis ,49. Chenet al. reportedet al.. Zhu et l.[et al. found thl.[et al. reportedl.[et al.. It was l.[et al.\u201358. Melal.[et al..It has been demonstrated that melatonin reduces flow shear stress-induced bone marrow mesenchymal stem cells injury by acting on melatonin receptors and the adenosine monophosphate-activated protein kinase/acetyl-CoA carboxylase signaling pathway \u25aa. In thiEarly experiments showed that treatment with melatonin can improve dyslipidemia . In patiIn conclusion, studies have demonstrated that melatonin has significant effects on ischemia-reperfusion injury, myocardial CIH injury, pulmonary hypertension, hypertension, vascular diseases, valvular heart diseases, and lipid metabolism Table . As an iH.S. wrote the manuscript, A.M.G. and S.Q. were involved in editing the manuscript.None.There are no conflicts of interest.\u25aa of special interest\u25aa\u25aa of outstanding interestPapers of particular interest, published within the annual period of review, have been highlighted as:"} +{"text": "Irs1\u2212/\u2212) are long-lived and enjoy a greater period of their life free from age-related pathology compared with wild-type (WT) controls. In this study, we show that primary dermal fibroblasts and primary myoblasts derived from Irs1\u2212/\u2212 mice are no more resistant to a range of oxidant and nonoxidant chemical stressors than cells derived from WT mice.Reduced signalling through the insulin/insulin-like growth factor-1 signalling (IIS) pathway is a highly conserved lifespan determinant in model organisms. The precise mechanism underlying the effects of the IIS on lifespan and health is currently unclear, although cellular stress resistance may be important. We have previously demonstrated that mice globally lacking insulin receptor substrate 1 ( Cytotoxicity was determined by calculating LD50 using probit analysis .We previously reported that gen Fig. or 3% oxgen Fig. . Indeed, O2 Fig. . We alsonce Fig. . Additionce Fig. , or in bnce Fig. were detIrs1\u2212/\u2212 mice are long-lived (Selman et al., et al., et al., et al., et al., Irs1\u2212/\u2212 mice occurs during development. Long-lived homozygous Chico flies, which lack the single Drosophila IRS protein (Clancy et al., Igf1R+/\u2212 mice (Kappeler et al., Irs1\u2212/\u2212 mice, unlike GH dwarfs, appear to have relatively normal somatotropic function and GH levels (Selman et al., et al., et al., et al., et al., Irs1\u2212/\u2212 mice are no more stress resistant than those derived from WT mice, suggesting that cellular stress resistance does not underlie the extended healthy lifespan of Irs1\u2212/\u2212 mice.While both male and female"} +{"text": "In the editorial section, Sulaiman Bah (134) discusses the possibility of dropping door-to-door enumeration in the next South African census. Priya Agrawal (135) points to persistent challenges in further reductions of maternal deaths in the United States of America.Gary Humphreys (138\u2013139) reports on the introduction of digital bidding to medicine procurement processes in African countries. Fiona Fleck interviews Eric Goosby (140\u2013141), the United Nations Secretary General\u2019s Special Envoy on Tuberculosis, about efforts to deliver more effective services.Jessica Cohen et al. (142\u2013151) measure the effects of stocking rapid diagnostic tests in local drug shops.Wei Cheng et al. (152\u2013160) study the effects of adding cotrimoxazole prophylaxis to antiretroviral therapy for HIV.Li Li et al. (169\u2013175) examine laws designed to prevent childhood injuries.Xiangming Fang et al. (176\u2013185) establish national estimates of child maltreatment.Alejandra Z\u00fa\u00f1iga-Fajuri (199\u2013202) reviews a policy change designed to increase the number of organs available for transplant.Willemijn\u00a0LA Sch\u00e4fer et al. (161\u2013168) assess patient satisfaction with primary care services in 34 countries.Nguyen Thanh Tam et al. (186\u2013198) analyse thirty years\u2019 of evidence on informed consent in clinical trials.Rosemary Wyber et al. (203\u2013208) describe potential health-sector use of large quantities of data generated for other purposes."} +{"text": "In the editorial section, Anthony Costello et al. (406) announce WHO\u2019s collaborative effort to define the syndrome associated with congenital Zika virus infection. Islene Araujo de Carvalho et al. (407) call for papers for a special issue on innovation for healthy ageing. Michel Sidib\u00e9 (408) points to an opportunity to secure funding for the next stage of the response to the human immunodeficiency virus epidemic.Menelaos Tzafalias (411\u2013412) reports from Greece on the fake lifejackets sold to refugees for their sea crossings to Europe. Richard Peto tells Andr\u00e9ia Azevedo Soares (413\u2013414) why WHO should focus on the big causes of death from noncommunicable diseases.Andre R Verani et al. (415\u2013423) review legislation designed to reduce tuberculosis transmission.Ifedayo MO Adetifa et al. (433\u2013441) report tuberculosis prevalence from a nationwide survey.Maureen O\u2019Leary et al. (442\u2013451) investigate delays in vaccination of low-birth-weight infants. Emmanuel Chanda et al. (475\u2013480) describe efforts to scale up vector control programmes.Ileana Heredia-Pi et al. (452\u2013461) measure the adequacy of antenatal care.Kelly L Edmunds et al. (424\u2013432) present ways to deal with contaminated waste..Kalipso Chalkidou et al. (462\u2013467) argue that explicit ways of setting priorities are neededIan Anderson et al. (468\u2013474) list the persuasive skills needed by health ministries."} +{"text": "Ramon Cajal, Marie Curie and Louis Pasteur among others had such qualities; they patiently pursued a flawless methodology which eventually led to success did (Ren\u00e9 Le Fort (1869-1951), derived his classic classification of facial fracture patterns by observing fractures arising from forceful blows subjected to the facial skeleton (Myron Firth Metzenbaum (1876-1944) (Alfred Washington Adson (1887\u20131951) (Howard Atwood Kelly (Keeping an open mind entices creativity; new concepts and research; ideas may then come right out-of-the-blue. Numerous scientists had undoubtedly seen a ripened apple fall from a tree before Newton 162 \u20131727 dt 1869-191, derive"} +{"text": "In the editorial section David Clarke et al. (482) explain that many countries need to revise their laws to support universal health coverage. Gretchen A Stevens et al. (483) introduce new reporting guidelines for health estimates. Rosanna Baker et al. (484) provide young people with ways to promote understanding of disease risks. \u00a0Patralekha Chatterjee (487\u2013488) reports on attempts made in India to improve air quality. \u00a0Fiona Fleck (489\u2013490) interviews Jeffrey D Sachs on the importance of accounting for the health impacts of energy policies.Davies Adeloye et al. (510\u2013521) count the burden of road traffic crashes, injuries and deaths.Samuel Oji Oti et al. (501\u2013509) study hypertension diagnosis and treatment in the community.Henry B Perry et al. (551\u2013553) use the Ebola response to argue that community health workers can help build resilient health systems.Tara M Tancred et al. (491\u2013500) recommend training in health policy and systems research.Anthony Danso-Appiah et al. (522\u2013533) review the evidence for accuracy of a diagnostic test.G\u00e9raldine Marks-Sultan et al. (534\u2013539) make a case for creating national public health laws where none exist.Vivica I Kraak et al. (540\u2013548) tally the progress made achieved in restricting marketing to children.Rita Kabra et al. (549\u2013550) report on a new approach from the programme on human reproduction. Aaron L Berkowitz (554\u2013556) calls for treatment protocols in situations where ischaemic and haemorrhagic strokes cannot be reliably differentiated."} +{"text": "In the editorial section, Peter Ventevogel et al. (666) call for the provision of mental health care in humanitarian emergencies. Brien A Holden et al. (667) emphasize the importance of better global estimates of presbyopia.In the news section, Dale Gavlak reports on health workers\u2019 struggles to help civilians caught up in the conflict in Yemen (670\u2013671). Rodrigo Guerrero-Velasco tells Alyssa Greenhouse how violence can be understood and prevented using an epidemiological approach (672-673). N\u00faria Homedes & Antonio Ugalde (674\u2013683) study the availability of new medicines in the countries where they were tested.Fen Wu et al. (684\u2013692) estimate deaths due to betel quid use.Zahirah McNatt et al. (719\u2013726) describe a national system for assessing the quality of facility-based care.Providing better mortality dataFaris Dababneh et al. (727\u2013731) review the progress made in national mortality surveillance and reporting.Somwe Wa Somwe et al. (732-736) describe a public-private partnership designed to improve the recognition and treatment of asthma. Lisa Eckenwiler et al. (737\u2013738) discuss the unintended health impacts of counterterrorism policies and practices. Meaghann S Weaver et al. (700\u2013711) analyse the design, delivery and outcomes of interventions to improve treatment adherence in low-resource settings. Corrina Moucheraud et al. (693\u2013699) evaluate the quality of economic data in applications to the WHO Model List of Essential Medicines.Kebede Deribe et al. (712-718) propose case definitions and elimination targets for endemic non-filarial elephantiasis.Molly Skerker et al. (739-740) describe obstacles to providing care to incarcerated pregnant women."} +{"text": "Howevelab, 2013; Reif, 2lab, 2013; Stewartlab, 2013). in vitro testing, namely in silico tissues . Typicaal., 2010; H\u00f6hme eal., 2010). In theal., 2010). For thal., 2010; Hoehme al., 2010). In nexal., 2010) have inIn toxicology, modelling especially structure activity and physiologically-based-pharmacokinetic (PBPK) models have a long standing tradition (Schug et al., 2013; Karaman"} +{"text": "In the editorial section, Jamie Bartram et al. (210) highlight a recent survey of 54 countries showing that thousands of health facilities do not have water, soap and/or toilets. Haik Nikogosian and Vera Luiza da Costa e Silva (211) mark the tenth anniversary of WHO's first global health treaty \u2013 the Framework Convention on Tobacco Control.Keiji Fukuda (212) describes the focus of this year\u2019s World Health Day: food safety. In an interview, Antoine Andremont (217\u2013218) focuses on the intersection of measures to improve food safety and decrease antimicrobial resistance.Fiona Fleck (215\u2013216) reports on efforts to get accurate tests for the Ebola virus closer to the point-of-care.Fabrice Quet et al. (219\u2013227) survey prescribing behaviours of doctors working in public hospitals.Claudia Diaz Olavarrieta et al. (249\u2013258) compare nurses\u2019 and doctors\u2019 provision of medical abortion services.Leonard\u00a0EG Mboera et al. (271\u2013278) describe lessons learnt from monitoring the national health sector plan.Nimalan Arinaminpathy\u00a0et al. (237\u2013248) study the Global Drug Facility\u2019s impact on tuberculosis medicine prices over the last decade. Kaspars Lunte et al. (279\u2013282) track recent price reductions for second-line treatments.\u00c9tienne V Langois et al. (259\u2013270) review the evidence for equitable access to postnatal care services.Nicola Magrini et al. (283\u2013284) outline challenges facing the next essential medicines committee.Scott\u00a0A McDonald et al. (228\u2013236) propose methods for getting from global to national burden of disease estimates"} +{"text": "In the editorial section, Tim Jinks et al. (558) explain why a global response is needed to confront antimicrobial resistance. Melanie\u00a0M Taylor et al. (559) present global estimates of the penicillin needed to prevent congenital syphilis. Wolbachia-infected mosquitoes in Brazil. Rabih El Chammay (564\u2013565) tells Fiona Fleck how the Lebanese health system is improving access to mental health care services.Andr\u00e9ia Azevedo Soares (562\u2013563) reports on the introduction of Beata \u015awi\u0105tkowska et al. (599\u2013604) report on efforts to provide care for people who worked in asbestos-processing plants.Shona Dalal et al. (613\u2013621) summarize reports of tetanus deaths and argue for renewed attention to vaccinating adolescent and adult men.Fathima Shihana et al. (622\u2013625) describe a bedside test used to recognize and manage methemoglobinemia.Ana Roberta Pati Pascom et al. (626\u2013630) evaluate the provision of peer-administered HIV tests.David Stuckler et al. (566\u2013573) analyse versions of WHO\u2019s sugars guidelines.R\u00e9mi N Charrel et al. (574\u2013584) summarize the information needed for diagnostic test development.Lacey LaGrone et al. (585\u2013598) review the evidence for references to WHO\u2019s trauma guidelines.Lisa G Johnston et al. (605\u2013612) insist on the need to check HIV status.LM Neufeld et al. (631\u2013632) call for more evidence to inform nutrition policies."} +{"text": "In the editorial section, Joseph Kutzin & Susan P Sparkes (2) define universal health coverage, health systems strengthening and health security and resilience. Samy A Alsirafy & Dina E Farag (3) argue that immediate action is needed to make oral morphine available in Egypt. In the news section, Dale Gavlak reports on how mental health services are becoming more available to the Syrian population in spite of the crisis (6\u20137). Charles Mgone talks to Fiona Fleck about building research partnerships (8\u20139).Leonor Maria Pacheco Santos et al. (22\u201329) assess if folic acid fortification has contributed to a reduction in neural tube defects. Hamado Ouedraogo et al. (37\u201345) evaluate the impact of a decade of preventive chemotherapy in school-age children. Shiwei Liu et al. (46\u201357) describe the creation of a national system for mortality surveillance. Mamta Gupta et al. (10\u201321) consider whether the civil registration system can be used as an accurate source of mortality data. Mark Zimmerman et al. (65\u201370) describe a staff support programme in rural hospitals.Thinakorn Noree et al. (30\u201336) investigate the economic impact of medical tourism.Simon Hales et al. (58\u201364) propose reporting guidelines for implementation and operational research. Jicui Dong & Zafar Mirza (71\u201372) encourage international development partners to foster local pharmaceutical production. Alfred Papali (73\u201374) uses the international space station to stress the feasibility of providing healthcare in remote areas."} +{"text": "Sendai framework for disaster risk reduction.In an editorial, Amina Aitsi-Selmi & Virgina Murray (362) introduce the In the news section, Patrick Adams and Fiona Fleck report on efforts to provide health information in languages other than English (365\u2013366). Jeremy Farrar talks to Fiona Fleck about the Wellcome Trust\u2019s priorities for research funding (367\u2013368).Agnes Binagwaho et al. (429\u2013434) describe efforts to coordinate eye-care services nationwide.Emma R Allanson & Robert C Pattinson (424\u2013428) study the associations between quality-of-care audits and perinatal mortality.Xiaolin Wei et al. (407\u2013416) measure patients\u2019 perceptions of primary care in Shanghai and Shenzhen.Ulrika Baker et al. (380\u2013389) track implementation bottlenecks.21Aaron Reeves et al. (369\u2013379) model the effects of economic recession on tuberculosis control.Medea Gegia et al. (390\u2013399) track the influence of smoking on tuberculosis treatment outcomes. Janine M Duke et al. (400\u2013406) compare long-term outcomes of burn survivors to the non-injured adult population. Ryota Sakamoto (435\u2013436) argues that legionellosis is one of the diseases influenced by climate.Emilie J Calvello et al. (417\u2013423) apply lessons learnt from obstetric emergencies."} +{"text": "AbstractAn updated checklist of the Birds of the Azores is presented based on information compiled from The checklist has a total of 414 species, including 38 new species.Almost half of the species and subspecies that occur in the Azores have a Palearctic origin, the remaining ones being essentialy Nearctic and Holarctic species. S\u00e3o Miguel is the island with the highest number of bird species, followed by Terceira, Corvo and Flores islands. Vertebrata: Aves) are some of the most iconic animals. They play important roles in the ecosystem, and since they are abundant and diverse in most urban and rural areas, humans have established a good long-lasting relationship with them . However, amongst the ca 10.000 bird species which have been living on Earth since the appearance of modern humans, many species were regionally lost or extinct, or are endangered, especially on islands Fig. , and Monhttp://www.birdingazores.com/), AdA \u2013 http://azores.avesdeportugal.info/) and ABS \u2013 http://azoresbs.weebly.com/ from January 2012 to February 2014 and http://azoresbirdsightings.blogspot.co.uk/ from August 2014 onward). These websites are presently used by birdwatchers as a tool to register new information about species occurrence and distribution in the Azores.http://www.worldbirdnames.org/). Some additional notes on the distribution of the different species among the islands and their biogeographic origin are presented.The present paper updates the previous checklist , with reBased on In order to make the data easier to consult, the main checklist (Checklist 1) includes all the breeding and non-breeding species together, including the escapes and/or introduced species which already have feral populations in the Azores. All the other escapes and/or introduced species were excluded from the main list, and can be consulted in Checklist 2.We considered the species with five our less records, in Azores, as Rare Species. This species are presented in a table and which is known to breed only on Praia and Baixo islets (Graciosa Island).Besides the taxonomical information and the distribution among the Azorean islands Fig. , we inclNearcticCOR; FLO; GRA; TER*; SMGOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758HolarcticA-IIA; AEWACOR; FLO; FAI; PIC; SJG; TER; SMG; SMR*Regular Wintering. Rodrigues et al. (2010)Gmelin, 1789NearcticCOR; FLO; FAI; PIC; SJG; TER; SMG; SMR*Regular Wintering. Rodrigues et al. (2010)Gmelin, 1789NearcticCOR; FLO; FAI; PIC; TER; SMG; SMR*Regular Wintering. Rodrigues et al. (2010)Linnaeus, 1758HolarcticA-IIA; AEWACOR*; FLO; FAI; TER; SMG; SMR*Regular Wintering. Rodrigues et al. (2010)Linnaeus, 1758PalearcticA-IIA; AEWACOR; FLO; FAI; PIC; GRA*; SJG; TER; SMG; SMRRegular Wintering. Rodrigues et al. (2010)Linnaeus, 1766NearcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Wintering. Rodrigues et al. (2010)Linnaeus, 1758PalearcticA-IIA; AEWACOR; FLO; FAI*; PIC; SJG*; TER; SMG; SMRRegular Wintering. Rodrigues et al. (2010)Linnaeus, 1758HolarcticA-IIA; AEWACOR ; FLO ; FAI; PIC; GRA*; SJG; TER; SMG ; SMRRegular Wintering. Rodrigues et al. (2010)Linnaeus, 1758PalearcticA-IIA; AEWACOR; FLO; FAI; PIC; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)Brewster, 1902NearcticCOR; FLO ; TER; SMGRegular Wintering. Rodrigues et al. (2010)Linnaeus, 1758HolarcticA-IIA; AEWACOR; FLO; FAI; TER; SMGOccasional Wintering. Rodrigues et al. (2010)HolarcticPIC*; TER; SMG; SMROccasional Wintering. Rodrigues et al. (2010)PalearcticCOR; TER*; SMG; SMROccasional Wintering. Rodrigues et al. (2010)Baillon, 1834PalearcticFLO; PIC*; TER; SMG; SMROccasional Wintering. Rodrigues et al. (2010)PalearcticCOR; TEROccasional Wintering. Rodrigues et al. (2010)NearcticFLO; FAI; PIC*; TER; SMG; SMR*Occasional Wintering. Rodrigues et al. (2010)NearcticACOR; FLO; FAI; PIC; SJG; TER; SMG; SMRRegular Wintering. Rodrigues et al. (2010)PalearcticCOR; FLO; TER; SMGOccasional Wintering. Rodrigues et al. (2010)PalearcticCOR; FLO; FAI; PIC; SJG*; TER; SMG; SMRRegular Wintering. Rodrigues et al. (2010)HolarcticCOR; FLO; FAI*; PIC*; SJG*; TER; SMG; SMROccasional Wintering. Rodrigues et al. (2010)PalearcticSMGOccasional Wintering. Rodrigues et al. (2010)HolarcticCOR; FLO; TER; SMGOccasional Wintering. Rodrigues et al. (2010)NearcticCOR*; FAI*; PIC*; GRA*; SJG; TER; SMG; SMR*Occasional Wintering. Rodrigues et al. (2010)PalearcticCOR*; FLO; SJG; SMGOccasional Wintering. Rodrigues et al. (2010)NearcticPIC; GRA; TEROccasional Migrant. Rodrigues et al. (2010)HolarcticCOR*; SJG; TER; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticTER; SMGOccasional Wintering. Rodrigues et al. (2010)HolarcticFAI*; PIC*; SJG*; TER; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticFAI; PIC; GRA; TER; SMGOccasional Migrant. Rodrigues et al. (2010)Afro-tropicalSMGOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; SMGOccasional Wintering. Rodrigues et al. (2010)HolarcticFLO; FAI; TER; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticFLO; FAI; SJG; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758HolarcticGRA; TER*Occasional Wintering. Rodrigues et al. (2010)Linnaeus, 1758HolarcticCOR; FLO; FAI; PIC; TER; SMG; SMROccasional Wintering. Rodrigues et al. (2010)NearcticFLO; TEROccasional Migrant. Rodrigues et al. (2010)HolarcticCOR; SMGOccasional Wintering. Rodrigues et al. (2010)HolarcticSMGOccasional Wintering. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)PalearcticFLO; TER*; SMGOccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)PalearcticA-IIAPIC (Breeder); GRA*; TER (Breeder); SMG*; SMR (Breeder)Introduced. Rodrigues et al. (2010)Hartert, 1917PalearcticA-IIBCOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Native. Rodrigues et al. (2010)PalearcticSMROccasional Migrant. New Azores RecordHolarcticCOR*; FLO; FAI; PIC; GRA; TER; SMG; SMRRegular Wintering. Rodrigues et al. (2010)HolarcticGRA; SMGOccasional Wintering. Rodrigues et al. (2010)Sub-AntarcticFAI; PICOccasional Migrant. Rodrigues et al. (2010)PantropicalP; A-I; B-II; T100FLO*; FAI*; PIC*; GRA*; SMG*; SMR (Breeder)Native. Rodrigues et al. (2010)PalearcticP; A-I; B-II; T100COR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Native. Rodrigues et al. (2010)HolarcticCOR; FLO; FAI; PIC; GRA*; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PantropicalCOR; FLO*; FAI; PIC; GRA*Occasional Migrant. Rodrigues et al. (2010)NearcticSMROccasional and Non-Breeding. Rodrigues et al. (2010)Mathews, 1934PalearcticGRAOccasional Migrant. New Azores Record PalearcticP; A-I; B-IICOR; FLO; FAI; PIC; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticFAI*; PIC; GRAOccasional Migrant. Rodrigues et al. (2010)Mathews, 1934PalearcticGRAOccasional Migrant. New Azores RecordPantropicalFAIOccasional Migrant. Rodrigues et al. (2010)PalearcticP; A-I; B-II; O; T100COR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER*; SMG (Breeder); SMR (Breeder)Macaronesian Endemic. Rodrigues et al. (2010)Southern AtlanticACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)Southern hemisphereACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)Lowe, 1921PalearcticFLO; FAI; SMGOccasional Migrant. Rodrigues et al. (2010)HolarcticP; A; B-IICOR (Breeder); FLO (Breeder); FAI*; PIC*; GRA*; SJG*; TER*; SMG (Breeder); SMR (Breeder)Native. Rodrigues et al. (2010)Sub-Antarctic and AntarcticCOR*; FLO; FAI*; PIC; GRA*; TER*; SMG; SMR*Regular Migrant. Rodrigues et al. (2010)PantropicalP; A-I; T100COR*; GRA (Breeder); SJG*; SMG*; SMR (Breeder)Native. Rodrigues et al. (2010)HolarcticCOR*; FLO; FAI; PIC; GRA; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)Sub-tropical PacificGRAOccasional Migrant. New Azores RecordBolton et al., 2008PalearcticP; A-I (x)COR*; GRA (Breeder); SJG*Azores Endemic. Rodrigues et al. (2010)PalearcticAFLO; FAI*; PIC; SMROccasional Migrant. Rodrigues et al. (2010)HolarcticFLO; TER; SMG; SMR*Occasional Wintering. Rodrigues et al. (2010)PalearcticSMGOccasional Wintering. Rodrigues et al. (2010)HolarcticFAIOccasional Wintering. New Azores RecordBrehm, 1831HolarcticFAI; TER; SMGOccasional Wintering. Rodrigues et al. (2010)NearcticCOR; FLO; PIC; TER; SMG; SMR*Occasional Wintering. Rodrigues et al. (2010)PalearcticFLO; TER*; SMGOccasional Wintering. Rodrigues et al. (2010)Pallas, 1811PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)Peters, 1930PantropicalA; AEWAFAI; PIC; GRA ; SMG*Occasional Migrant. Rodrigues et al. (2010)Daudin, 1802PantropicalCOR*; FLO; FAI*; SJG*; TER*; SMG*Occasional Migrant. Rodrigues et al. (2010)PalearcticPIC*; TEROccasional Migrant. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR; FLO; PIC; TER; SMG; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)HolarcticCOR*; FLO*; FAI*; PIC*; GRA*; SJG*; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticSMROccasional Migrant. Rodrigues et al. (2010)Gmelin, 1789NearcticCOR; FLO; FAI; PIC; GRA*; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticA; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)Linnaeus, 1758NearcticCOR; FLO; FAI; PIC; SJG; TER; SMGOccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)Linnaeus, 1766PalearcticCOR*; PIC*; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)PalearcticCOR*; FAI*; PIC; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticCOR*; FLO; PIC*; GRA*; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticTER; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticA; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Wintering. Rodrigues et al. (2010)NearcticFLO; PIC; SJG; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticFLO; PIC; GRA*; SJG; SMG*Occasional Migrant. Rodrigues et al. (2010)PalearcticP; A-I; B-II; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)NearcticFLO; FAI; PIC*; GRA*; SJG; TER; SMGOccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticPIC; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticTER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticFLO; PIC; GRA; SJG; TER; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; PIC; TER; SMROccasional Wintering. New Azores RecordHolarcticCOR; FLO; FAI; PIC; GRA; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Mathews, 1914Sub-tropical and Tropical Atlantic; Tropical-eastern PacificCOR*; FLO*; PIC*; TER*; SMGOccasional Migrant. Rodrigues et al. (2010)HolarcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)Lesson, 1831PantropicalFAI; GRA*Occasional Migrant. Rodrigues et al. (2010)PantropicalGRA*; SMGOccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; PIC; GRA*; TER; SMG; SMROccasional Wintering. Rodrigues et al. (2010)HolarcticCOR; PIC*; GRA*; TER; SMG; SMROccasional Wintering. Rodrigues et al. (2010)HolarcticCOR*; FLO; FAI; PIC; TER; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticP; A; B-II; T100COR; FLO; FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Azores Endemic. Rodrigues et al. (2010)PalearcticGRA; TEROccasional Migrant. New Azores RecordHolarcticCOR; FAI; PIC*; GRA*; TEROccasional Migrant. Rodrigues et al. (2010)NearcticCOR*; FLO; TER*Occasional Migrant. Rodrigues et al. (2010)PalearcticFLO; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO; PIC*; TER; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)PalearcticTEROccasional Migrant. Rodrigues et al. (2010)NearcticFLO; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticGRA; SMROccasional Migrant. New Azores RecordPalearcticCOR; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FAI; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Gmelin, 1789NearcticFLO; FAI*; PIC; GRA*; TER; SMGOccasional Wintering. Rodrigues et al. (2010)Linnaeus, 1758PalearcticA-IIA; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER (Breeder); SMG (Breeder); SMRRegular Wintering. Rodrigues et al. (2010)PalearcticA-IIB; AEWACOR*; FLO (Breeder); FAI*; PIC*; GRA*; SJG*; TER (Breeder); SMG (Breeder); SMR (Breeder)Native. Rodrigues et al. (2010)Thomson, 1842PalearcticGRA*; TER*; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticFLO; FAI*; GRA*; SMGOccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticFLO*; SJG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticFAI; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO*; PIC*; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticTER; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticFLOOccasional Migrant. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticFAI; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticPIC; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticSMG; SMR*Occasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticP; A-I; B-II; AEWACOR; FLO; FAI; PIC; GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Regular Wintering. Rodrigues et al. (2010)Pallas, 1773PalearcticCOROccasional Migrant. New Azores RecordScopoli, 1786PalearcticA; B-II; AEWACOR; FLO; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758HolarcticA; B-II; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR; FAI; SMROccasional Migrant. Rodrigues et al. (2010)Bonaparte, 1825NearcticACOR; FLO; FAI; PIC; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)Linnaeus, 1758NearcticCOR; FLO; FAI; PIC*; TER; SMG; SMR* Occasional Migrant. Rodrigues et al. (2010)PalearcticCOR*; FLO; FAI; GRA; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; PIC; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)HolarcticTER; SMR*Occasional Migrant. Rodrigues et al. (2010)HolarcticA-IIB; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)PalearcticA-IIB; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMROccasional Wintering. Rodrigues et al. (2010)PalearcticCOR; FLO; FAI; PIC; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)HolarcticA; B-II; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)NearcticCOR; FLO; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticTEROccasional Migrant. Rodrigues et al. (2010)HolarcticA; B-II; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)HolarcticP; A-I; B-II; AEWAFLO; FAI; PIC; GRA; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)HolarcticFLO*; SJG; TER; SMGOccasional Migrant. Rodrigues et al. (2010)HolarcticA-IIB; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)PalearcticA; B-II; AEWACOR; FLO; FAI; PIC; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticACOR; FLO; FAI; PIC; GRA*; SJG; TER; SMG; SMR*Regular Migrante; Occasional Wintering. Rodrigues et al. (2010)NearcticPIC; SJG*; SMGOccasional Migrant. Rodrigues et al. (2010)HolarcticCOR; FLO; FAI; PIC; GRA; TER; SMGOccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticCOR*; PIC; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)HolarcticCOR; FLO; FAI; PIC; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO; FAI; PIC; SJG; TER; SMG; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; PIC; SJG; TER; SMGOccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; PIC; GRA*; SJG; TER; SMG; SMR*Regular Migrante; Occasional Wintering. Rodrigues et al. (2010)PalearcticTER; SMG*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; FAI*; PIC; GRA; SJG; TER; SMG; SMR*Regular Migrant. Rodrigues et al. (2010)PalearcticA-IIA; AEWACOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA*; SJG (Breeder); TER (Breeder); SMG (Breeder); SMR*Native. Rodrigues et al. (2010)PalearcticSMROccasional Migrant. New Azores RecordNearcticCOR; FLO; PIC; SJG; TER; SMG; SMR*Occasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticCOR; TER; SMG; SMR*Occasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticTEROccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO*; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)PalearcticA-IIB; AEWASJG; TER; SMGRegular Migrant. Rodrigues et al. (2010)PalearcticCOR; FAI; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticFLO*; PIC; GRA; TER; SMG; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)Latham, 1790NearcticCOR; FLO; FAI*; PIC*; GRA; SJG*; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)PalearcticA-IIB; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)Vieillot, 1817PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)HolarcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)HolarcticTER*; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; PIC; TER; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticP; A-I;A-IIB; AEWACOR; FLO; FAI; PIC; GRA*; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticA-IIA; AEWACOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA*; SJG (Breeder); TER (Breeder); SMG (Breeder); SMR*Native. Rodrigues et al. (2010)PalearcticFAI; TER; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMR*Regular Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticFLO; PIC*; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; PIC; TER; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticA-IIB; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticFLO; PIC; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticSJG; TER; SMGOccasional Migrant. Rodrigues et al. (2010)Wilson, 1813NearcticCOR*; FLO; GRA*; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticFLO; PIC*; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticFLO; SJG; TER; SMG; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; PIC; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)HolarcticTEROccasional Wintering. New Azores RecordPantropicalFLOOccasional Migrant. Rodrigues et al. (2010)PalearcticFLO; TER; SMG*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; TER; SMG*Occasional Migrant. Rodrigues et al. (2010)PalearcticTER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticTER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)NearcticFAI; PIC; TER; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticA-IIB; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)HolarcticPIC; TER; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticFAI; SMG; SMROccasional Migrant. Rodrigues et al. (2010)HolarcticFLO; TER; SMG*Occasional Migrant. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)PalearcticFAI; PIC*; TER; SMGOccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)Pontoppidan, 1763PalearcticFLO; FAI; TER; SMGOccasional Migrant. Rodrigues et al. (2010)Richardson, 1831NearcticTEROccasional Wintering. Rodrigues et al. (2010)PalearcticCOR; FLO; PIC; TER; SMGOccasional Wintering. Rodrigues et al. (2010)Ord, 1815NearcticACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Wintering. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)Meyer, 1822HolarcticCOR; FLO; FAI; PIC; GRA; TER; SMGOccasional Wintering. Rodrigues et al. (2010)Brewster, 1883NearcticFLO; TER*; SMG; SMR*Occasional Wintering. Rodrigues et al. (2010)Gunnerus, 1767HolarcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Wintering. Rodrigues et al. (2010)Linnaeus, 1758HolarcticA-IIB; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Wintering. Rodrigues et al. (2010)Dwight, 1922PalearcticP; A-IIB; AEWACOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Azores Endemic. Rodrigues et al. (2010)Naumann, 1840PalearcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Coues, 1862NearcticCOR*; FLO; FAI; PIC; SJG*; TER; SMGOccasional Wintering. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; PIC; GRA; TER; SMG; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticFLO; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)PantropicalPIC; GRA ; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Tropical AtlanticA; AEWAFAI*; GRA (Breeder); SJG*; TER*; SMG*; SMRNative. Rodrigues et al. (2010)HolarcticA; AEWACOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant; Regular Wintering. Rodrigues et al. (2010)Montagu, 1813HolarcticP; A-I; B-II; AEWA; O; T100COR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Native. Rodrigues et al. (2010)Nuttall, 1834NearcticCOR; FLO; TER; SMG*Occasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758HolarcticP; A-I; B-II; AEWA; T100COR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Native. Rodrigues et al. (2010)Pontoppidan, 1763HolarcticCOR; FLO; FAI; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)PalearcticFLO; SMGOccasional Migrant. Rodrigues et al. (2010)HolarcticFLO; GRA; TEROccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMGOccasional Migrant. Rodrigues et al. (2010)HolarcticCOR*; FLO; FAI; PIC*; GRA; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Vieillot, 1819HolarcticFAI; PIC; GRA*; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Saunders, 1893AntarcticFLO*; FAI*; GRA*; SMGOccasional Migrant. Rodrigues et al. (2010)HolarcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)HolarcticCOR; FLO; FAI; PIC; GRA; SJG*; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMRRegular Migrant. Rodrigues et al. (2010)Linnaeus, 1758HolarcticFAI; TER; SMGOccasional Wintering. Rodrigues et al. (2010)PalearcticCOR; FAI; PIC; GRA; SJG; TER; SMGOccasional Wintering. Rodrigues et al. (2010)HolarcticFLO; FAI; PIC; GRA*; TER; SMG; SMROccasional Wintering. Rodrigues et al. (2010)HolarcticFLO*; FAI; PIC*; TER; SMGOccasional Wintering. Rodrigues et al. (2010)Gmelin, 1789PalearcticA-IIACOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Introduced. Rodrigues et al. (2010)Hartert, 1905PalearcticP; A-ICOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Azores Endemic. Rodrigues et al. (2010)PalearcticCOR*; FAI*; PIC (Breeder); GRA*; TER (Breeder); SMG; SMRNative. Rodrigues et al. (2010)PalearcticCOR; FLO; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLOOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticCOR*; FLO; SMGOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticPIC*; SJG; TER*; SMG; SMROccasional Migrant. Rodrigues et al. (2010)HolarcticSMGOccasional Migrant. Rodrigues et al. (2010)HolarcticCOR; FLO; FAI; PIC; GRA; SJG*; TER; SMG; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)PalearcticA; B-IICOR*; FLO*; FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR*Native. Rodrigues et al. (2010)HolarcticCOR*; FLO; FAI; PIC*; SJG*Occasional Wintering. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR*; GRA*; TER; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; PIC; SJG; TER; SMGOccasional Migrant. Rodrigues et al. (2010)Afro-tropicalSMROccasional Migrant. Rodrigues et al. (2010)PalearcticFLO; FAI; GRA; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticFLO*; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; FAI; TER; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FAI*; TER*; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticSMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticSMROccasional Migrant. Rodrigues et al. (2010)NearcticFLO; FAI; PIC; GRA; SJG; TER*; SMR*Occasional Migrant; Occasional Wintering. Rodrigues et al. (2010)Linnaeus, 1758PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticFLO; PIC*; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FAIOccasional Migrant. New Azores RecordNearcticCOROccasional Migrant. Rodrigues et al. (2010)Radde, 1863PalearcticPICOccasional Migrant. New Azores RecordLinnaeus, 1758HolarcticCOR; FLO; TER; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)G\u00e9n\u00e9, 1839PalearcticSMGOccasional Migrant. New Azores RecordFleischer, 1818PalearcticSMG; SMR*Occasional Migrant. Rodrigues et al. (2010)Tunstall, 1771HolarcticCOR; FLO; FAI; PIC*; GRA; TER; SMG; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)Linnaeus, 1758HolarcticAZOOccasional Migrant. New Azores RecordLinnaeus, 1758NearcticTER; SMGOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR; SMG*Occasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR; FLO; FAI; PIC; GRA; TER; SMG; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)Linnaeus, 1766PalearcticCOR*; FLO; FAI; PIC; TER*; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)Afro-tropicalFAI*; TER*; SMG (Breeder)Introduced. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOROccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticSMG; SMROccasional Migrant. New Azores RecordVieillot, 1808NearcticCOROccasional Migrant. New Azores RecordVieillot, 1808NearcticCOR; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; FAI*; TER*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO*Occasional Migrant. Rodrigues et al. (2010)PalearcticCOR*; FLO; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticPIC; TER; SMGOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticSMG; SMROccasional Migrant. Rodrigues et al. (2010)Vieillot, 1808NearcticCOR; FLOOccasional Migrant. New Azores RecordHolarcticFAIOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR; FLO; FAI; TER*; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticSJG; SMR*Occasional Migrant. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)PalearcticCOR*; FAI; SJG; SMG*Occasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO; FAI; PIC; GRA; SJG*; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Boddaert, 1783NearcticCOR; FLO; TEROccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR; FLO; FAI; PIC*; GRA; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; TEROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLOOccasional Migrant. Rodrigues et al. (2010)PalearcticSMROccasional Migrant. New Azores RecordHolarcticCOR*; FLO; PIC; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO; FAI; PIC; GRA*; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticPIC*; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO; SJG; TER*; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; TEROccasional Migrant. New Azores RecordPalearcticCOROccasional Migrant. Rodrigues et al. (2010)PalearcticFLOOccasional Migrant. New Azores RecordPalearcticCOROccasional Migrant. New Azores RecordSeebohm, 1883PalearcticP; A; B-IISMG (Breeder)Azores Endemic. Rodrigues et al. (2010)Murphy & Chapin, 1929PalearcticP; A; B-IIFLO; FAI; PIC; GRA; SJG; TERAzores Endemic. Rodrigues et al. (2010)Vaurie, 1954PalearcticP; A; B-IISMR (Breeder)Azores Endemic. Rodrigues et al. (2010)Alexander, 1898PalearcticA; B-IICOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Azores Endemic. Rodrigues et al. (2010)PalearcticCOR; FLO*Occasional Migrant. Rodrigues et al. (2010)PalearcticCOROccasional Migrant. New Azores RecordPalearcticFLOOccasional Migrant. New Azores RecordLatham, 1787PalearcticTEROccasional Migrant. New Azores RecordPalearcticSMGOccasional Migrant. Rodrigues et al. (2010)NearcticFLOOccasional Migrant. New Azores RecordHartert, 1903PalearcticA-IIBCOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Azores Endemic. Rodrigues et al. (2010)NearcticCOR; FLO*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLOOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLOOccasional Migrant. New Azores RecordNearcticCOR*; SMGOccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticSMG; SMROccasional Migrant. Rodrigues et al. (2010)Hartert, 1905PalearcticA-IIBCOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Azores Endemic. Rodrigues et al. (2010)Linnaeus, 1766NearcticCOROccasional Migrant. New Azores RecordTemminck, 1820PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)Brehm, 1831PalearcticSMG; SMROccasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR; FLO; PIC*; GRA; TER; SMG; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR; SMG*; SMR*Occasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)PalearcticA; B-IIFAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Native. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)PalearcticTEROccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO; SMROccasional Migrant. New Azores RecordNearcticCOR; FLO; FAI; PIC; GRA; SJG; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticA; B-IIFLO; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticFLOOccasional Migrant. Rodrigues et al. (2010)PalearcticTER*; SMG; SMROccasional Migrant. Rodrigues et al. (2010)PalearcticCOR* ; FLO; PIC; SMGOccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)PalearcticCOR*; FLO; SJG; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)PalearcticACOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Introduced. Rodrigues et al. (2010)PalearcticTER; SMGOccasional Migrant. Rodrigues et al. (2010)Afro-tropicalTER (Breeder); SMG (Breeder)Introduced. Rodrigues et al. (2010)PalearcticSMGOccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; TER*; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)PalearcticFLO; TEROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO*; PIC; TER; SMG; SMROccasional Wintering. Rodrigues et al. (2010)PalearcticCOR; TEROccasional Migrant. New Azores RecordLinnaeus, 1758PalearcticCOR*; FLO; GRA; TER; SMG; SMROccasional Migrant. Rodrigues et al. (2010)Vaurie, 1957PalearcticA; B-IICOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Azores Endemic. Rodrigues et al. (2010)Pallas, 1776PalearcticCOR; SMR*Occasional Migrant. Rodrigues et al. (2010)Linnaeus, 1758PalearcticCOR; PIC; TER*; SMG; SMROccasional Migrant. Rodrigues et al. (2010)HolarcticCOR; FLO; SMGOccasional Migrant. Rodrigues et al. (2010)HolarcticCOROccasional Migrant. Rodrigues et al. (2010)Tschusi, 1901PalearcticA; B-IICOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Macaronesian Endemic. Rodrigues et al. (2010)PalearcticA; B-IIFAI*; PIC*; TER (Breeder); SMG (Breeder); SMR*Introduced. Rodrigues et al. (2010)Pucheran, 1859PalearcticACOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Azores Endemic. Rodrigues et al. (2010)Linnaeus, 1758PalearcticSMROccasional Wintering. New Azores RecordPalearcticTER*; SMGOccasional Migrant. Rodrigues et al. (2010)PalearcticFLOOccasional Migrant. New Azores RecordLinnaeus, 1758HolarcticSMGOccasional Migrant. Rodrigues et al. (2010)Godman, 1866PalearcticP; A-I; T100SMG (Breeder)Azores Endemic. Rodrigues et al. (2010)PalearcticACOR (Breeder); FLO (Breeder); FAI (Breeder); PIC (Breeder); GRA (Breeder); SJG (Breeder); TER (Breeder); SMG (Breeder); SMR (Breeder)Macaronesian Endemic. Rodrigues et al. (2010)PalearcticTEROccasional Migrant. Rodrigues et al. (2010)PalearcticCOR; FLO; SMGOccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticCOROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLOOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR*; SMG*; SMROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO*; TEROccasional Migrant. Rodrigues et al. (2010)NearcticCOROccasional Migrant. Rodrigues et al. (2010)NearcticCOROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticCOROccasional Migrant. New Azores RecordNearcticCOROccasional Migrant. New Azores RecordNearcticCOR*; FLOOccasional Migrant. Rodrigues et al. (2010)NearcticCOROccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; SMGOccasional Migrant. Rodrigues et al. (2010)NearcticCOROccasional Migrant. Rodrigues et al. (2010)NearcticCOROccasional Migrant. New Azores RecordOlson & Reveal, 2009NearcticCOROccasional Migrant. New Azores RecordNearcticCOR; FLO; SMR*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; SMR*Occasional Migrant. Rodrigues et al. (2010)Pallas, 1776PalearcticSMROccasional Migrant. New Azores RecordNearcticFLOOccasional Migrant. Rodrigues et al. (2010)NearcticCOROccasional Migrant. New Azores RecordNearcticCOR; FLOOccasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLO; SMR*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; SMR*Occasional Migrant. New Azores RecordNearcticCOR; FLO; SMR*Occasional Migrant. Rodrigues et al. (2010)HolarcticCOR; FLO; TER; SMGOccasional Migrant. Rodrigues et al. (2010)HolarcticCOR; FLO; FAI; PIC; GRA; TER; SMG; SMROccasional Migrant; Occasional Wintering. Rodrigues et al. (2010)NearcticCOR; FLO; SMG; SMR*Occasional Migrant. Rodrigues et al. (2010)NearcticCOR; FLOOccasional Migrant. Rodrigues et al. (2010)NearcticCOROccasional Migrant. Rodrigues et al. (2010)NearcticCOR*; FLO; SMR*Occasional Migrant. Rodrigues et al. (2010)PalearcticPIC*; GRA; TER; SMGOccasional Migrant. Rodrigues et al. (2010)Afro-tropicalSMGIntroduced. Rodrigues et al. (2010)NearcticFAIIntroduced. Rodrigues et al. (2010)PalearcticA-IIASMGIntroduced. Rodrigues et al. (2010)Linnaeus, 1758PalearcticAZOIntroduced. Rodrigues et al. (2010)NearcticSMGIntroduced. Rodrigues et al. (2010)Afro-tropicalSMGIntroduced. Rodrigues et al. (2010)Afro-tropicalTERIntroduced. Rodrigues et al. (2010)The updated list of the bird species recorded in the Azores comprised 38 newly recorded species, increasing the total number of recorded species and subspecies to 414. These 414 taxa belong to 24 orders, 67 families, 202 genera and 404 species. Some of the species and genus names from the previous list were changed and some families were moved to other orders. The taxonomic revision results in the addition of four new orders to the checklist.S\u00e3o Miguel Island stands out as the richest island (299 spp. and sspp.), followed by Terceira (248 spp. and sspp.), Corvo and Flores , Charadriiformes and Anseriformes are by far the most represented orders with 129, 111 and 42 taxa, respectively.Almost half of the species and subspecies that occur in the Azores come from the Palearctic but the Nearctic is also well represented . The species and subspecies of Holarctic origin totalled 68 , but also an enlargement of the distribution of several species among the different Azorean islands. A simple explanation for this could be the increase in the number of observers performing birdwatching out of the \u201chigh season\u201d (autumn and spring) for vagrant birds, and possibly also the creation of the \u201cThe situation of the Azores in the middle of the North Atlantic makes these islands a safe place for the birds taken away from their normal migratory routes by strong winds or by changes in wind direction (vagrants) . VagrantThe native birds from the Azores provide good examples of insular speciation, since two endemic species and 11 endemic subspecies are known. Moreover, many more endemic terrestrial species have gone extinct , possiblSpecies lists such as this one are important to draw attention to particular taxonomic groups, to serve as a baseline in long-term systematic monitoring, to elucidate patterns of species diversity and distribution, to elucidate information gaps about distribution, and can serve as a tool in educational projects. The currently ongoing initiatives concerning citizen ornithological science in the Azores will be of high value for the knowledge of the biology and phenology of migratory birds and will hopefully increase the pressure towards the definition and preservation of more Important Bird Areas (IBAs) in the archipelago.Supplementary material 1Global species richness on the nine Azorean islandsData type: Species distributionBrief description: Global species richness on the nine Azorean islands .File: oo_62386.xlsxBarcelos et al.Supplementary material 2Biogeographical origin of the species present in Azores archipelagoData type: occurencesBrief description: Biogeographical origin of the species present in Azores archipelago.\u201cOther\u201d refers to marine regions only represented once .File: oo_62383.xlsxBarcelos et al.Supplementary material 3List of Rare SpeciesData type: occurencesBrief description: Species with five or less records and individuals counts, in the Azores, with some notes.File: oo_62653.xlsxBarcelos et al."} +{"text": "The Editor depends heavily on reviewers in forming opinions about papers submitted to Parasite.We are happy to formally thank the following scientists for their help, great effort and effectiveness in improving the quality of manuscripts.Since January 2013, the rejection rate of Parasite has been about 50%; this list includes reviewers who evaluated papers which were not published.For 2013 and 2014, our 292 reviewers from 51 countries are listed below. Names are in alphabetical order. Some have reviewed more than one manuscript.\u00a0Mohammd Abdigoudarzi (Iran)Rashad Abdul-Ghani (Egypt)Rob Adlard Peter H. Adler (United States)Sargis Aghayan (Armenia)Patrice Agnamey (FranceDaniel Ajzenberg (France)Fabio Akashi Hernandes (Brazil)Okan Akhan (Turkey)Sonia Almeria (Spain)Bulent Alten (Turkey)Alessandro Amarante (Brazil)Karim Aoun (Tunisia)Harry Archimede (France)Herbert Auer (Austria)Ali Ayadi (Tunisia)Mehmet Fatih Aydin (Turkey)Carlos Azevedo Fr\u00e9d\u00e9ric Baldacchino (France)Thomas Barth (Germany)Brigitte Bartholomot (France)Eva B\u00e1rtov\u00e1 (Czech Republic)Jean-Claude Beaucournu (France)Mustapha Benazzouz (Morocco)Rosa Ma. del Refugio Bermudez Cruz (Mexico)Fr\u00e9d\u00e9ric Beugnet (France)Ian Beveridge Sarah Bevins (United States)Bhen Sikina Toguebaye Pascal Boireau (France)Marta Bombarova (Slovakia)Sarah Bonnet (France)Christian Bories (France)Fran\u00e7oise Bouchet (France)Franck Bou\u00e9 (France)Nacim Bouheraoua (France)J\u00e9r\u00e9my Bouyer (France)Geoffrey Boxshall (United Kingdom)Rodney A. Bray (United Kingdom)Niel L. Bruce Magdal\u00e9na Brunansk\u00e1 (Slovakia)Jacques Cabaret (France)Simone Caccio Gioia Capelli Luis Cardoso Graca Casal Sibeli B. S. Cembranelli (Brazil)Gabriela Certad (France)Qijun Chen (China)Ren\u00e9 Chermette (France)Neil Chilton (Canada)Bruno B. Chomel (United States)Graham Clark (United Kingdom)Helena Collgros (Spain)Vince Connors (United States)Miriam F. Cooperband (United States)S. Z. Coskun (Turkey)Mark Costello Thomas Cribb Liwang Cui (United States)Bruno da Rocha Azevedo (United States)John Dalton (Canada)Marie-Laure Dard\u00e9 (France)Isaure de Buron (United States)Johan F. De Jonckheere (Belgium)Danielle Defaye (France)Gunita Deksne (Latvia)Pascal Delaunay (France)Laurence Delhaes (France)J\u00e9r\u00f4me Depaquit (France)Peter Deplazes (Switzerland)Anastasia Diakou (Greece)Dimitar Dimitrov (Bulgaria)Olgica Djurkovic (Serbia)Philippe Dorchies (France)Pierre Dorny (Belgium)Pascal Drakulovski (France)Gilles Dreyfuss (France)Michael Dryden (United States)Jitender P. Dubey (United States)Michael Duch\u00eane (Austria)Jean Dupouy-Camet (France)Lance Durden (United States)G\u00e9rard Duvallet (France)Chadli Dziri (Tunisia)Samar N. El-Beshbishi (Egypt)Agustin Estrada-Pena (Spain)Loic Favennec (France)Andy Fenton (United Kingdom)Ignacio Ferre P\u00e9rez (Spain)Raina Fichorova (United States)John Fleming (United States)Isabelle Florent (France)William Font (United States)Michel Franc (France)Montserrat Gallego (Spain)Martin Garcia Varela (Mexico)Lamia Gargouri (Tunisia)Claudia G\u00e9rard (France)Mohamed Gharbi (Tunisia)David I. Gibson (United Kingdom)C\u00e9cile Ginane (France)Patrick Giraudoux (France)Jorge Gomez-Marin (Colombia)Jorge F. Gonzalez (Spain)David Gonzalez-Solis (Mexico)Bruno Gottstein (Switzerland)Sophie Goyard (France)Katarzyna Go\u017adzik (Poland)Tilmann Graeter (Germany)Philippe Grellier (France)Fr\u00e9deric Grenouillet (France)Norman Grim (United States)Pascale Gueirard (France)Patrick Gu\u00e9rin (Switzerland)Neelima Gupta (India)Carlos Gutierrez (Spain)Hossein Hamidinejat (Iran)Patrick Hamilton (United Kingdom)Ralf Harbach (United Kingdom)Shawgi Hassan (Sudan)Michael Hastriter (United States)Hilda Hernandez (Cuba)Tony Holder (United Kingdom)Ren\u00e9 Houin (France)Sandrine Houz\u00e9 (France)Shaomin Hu (United States)Yiau-Min Huang (United States)Michael Huffman (United States)Tine Huyse (Belgium)Furhan Iqbal (Pakistan)Kunihiko Izawa (Japan)Yves Jackson (Switzerland)Kym C. Jacobson (United States)Philippe Jacquiet (France)Aaron R. Jex F. Agustin Jimenez (United States)Arlene Jones (United Kingdom)Frans Jongejan (Netherlands)Quentin Jossart (Belgium)Kerstin Junker (South Africa)Jean-Lou Justine (France)Sukhbir Kaur (India)Peter Kern (Germany)Jamal Khalife (France)Jean-Pierre Kinet (United States)Jenny Knapp (France)Janet Koprivnikar (Canada)Mustafa Kose (Turkey)Sabrina Krief (France)\u00c1rni Kristmundsson (Iceland)Delane Kritsky (United States)Roman Kuchta (Czech Republic)Katrin Kuhls (Germany)Sandrine Lacour (France)Kevin Lafferty (United States)Ir\u00e8ne Landau (France)Gordon Langsley (France)Wonchoel Lee (Korea)Nils Leine (Norway)Robert Lester Robert E. Lewis (United States)Jun Li (China)Marshall Lightowlers L. H. Susan Lim Renyong Lin (China)David Scott Lindsay (United States)Tim Littlewood (United Kingdom)Badre lmimouni (Morocco)Sean A. Locke (Canada)Philippe Loiseau (France)Bertrand Losson (Belgium)Helder Louvandini (Brazil)Liang Lu (China)Hugo D. Lujan (Argentina)Joseph W. Lynch Sutherland Maciver (United Kingdom)Charles Mackenzie (United States)Herv\u00e9 Maisonneuve (France)Laurence Malandrin (France)Vincent Mallet (France)Nathalie Mandonnet (France)Sven Mangelinckx (Belgium)Georges Mantion (France)Laurence Marchat (Mexico)Coralie Martin (France)Alvaro Martinez Moreno (Spain)Vesna Matijatko (Croatia)Sheena McGowan (United States)Concepta McManus (Brazil)Antonio Menezes da Silva Mercedes Mezo (Spain)Terry Miller Laurence Millon (France)Jordi Miquel (Spain)Gholamreza Molavi (Iran)Marcelo Molento (Brazil)Scott Monks (Mexico)Gisela R. A. Monteiro Marques (Brazil)Jorge Morales-Montor (Brazil)Serge Morand (France)Franti\u0161ek Moravec (Czech Republic)Sara Moutailler (France)Yasen Mutafchiev (United States)Lassad Neifar (Tunisia)C\u00e9dric Neveu (France)Anthony L. Newsome (United States)Violaine Nicolas (France)Karsten Noeckler (Germany)Emiliano Ocampo (Argentina)Lucia H. O\u2019Dwyer (Brazil)Annemarie Ohler (France)Faith Osier (United Kingdom)Domenico Otranto Harry Palm (Germany)Carine Paraud (France)Philippe Parola (France)Maria Paulke-Korinek (Austria)Adalberto P\u00e9rez de Leon (United States)Anton P\u00e9rez-Rodriguez (Spain)Bernard Pesson (France)Romualda Petkevi\u010di\u016bt\u0117 (Lithuania)Kurt Pfister (Germany)Davies Mubika Pfukenyi (Zimbabwe)Wojciech Piasecki (Poland)Robert Poulin Edoardo Pozio Heather C. Proctor (Canada)Eric Pulis (United States)Karl Marx Quiazon (Philippines)David Ranucci Christophe Ravel (France)Richard Reithinger (United States)Jean-Michel Rep\u00e9rant (France)Vincent Robert (France)Mike Rogan (United Kingdom)Jos\u00e9e Roy (Canada)Sadie F. Ryan (United States)Guilermo Salgado-Maldonado (Mexico)Rodrigo Sanabria (Argentina)Julien Santi-Rocca (France)Aur\u00e9lia Saraiva Brad Scandrett (Canada)Gereon Schares (Germany)Tom\u00e1\u0161 Scholz (Czech Republic)Rolf K. Schuster (Germany)Tengku Shahrul Anuar Jeffrey Shaw (Brazil)Andrew Shinn (United Kingdom)Didier Sicard (France)Anne Silvestre (France)Fernando Simon (Spain)Bibhuti Singh (United States)George Snounou (France)Smaragda Sotiraki (Greece)Isaia Sotiriadou (Germany)Sabine Specht (Germany)David Spratt Amir Steinman (Israel)Balazs Szoor (United Kingdom)Rachida Tahar (France)Dennis Tappe (Germany)David Taylor (United States)Thomas H. Terrill (United States)Richard Thacker (United Kingdom)Jean-Marc Thellier (France)Dave J. Thornton (United Kingdom)Paul Torgerson (Switzerland)Carine Truyens (Belgium)Gerrit Uilenberg (France)Gediminas Valkiunas (Lithuania)Thirumalaisamy P. Velavan (Germany)Veerle Versteirt (Belgium)Isabelle Villena (France)Christian Vivar\u00e8s (France)Petr Volf (Czech Republic)Dominique A. Vuitton (France)Martine Wallon (France)Jianyang Wang (United States)Andrew Williams (Denmark)Thierry Wirth (France)Anne-Sophie Woronoff (France)Lihua Xiao (United States)Cecilia Ximenez (Mexico)Na Yang (China)H\u00e9l\u00e8ne Yera (France)Timothy Yoshino (United States)Arnaldo Zaha (Brazil)Eberhard Zeyhle (Kenya)Yanlin Zhao (United States)Xing-Quan Zhu (China)Zhizhang Zhuang (China)"} +{"text": "In the editorial section, Margaret Chan (818) argues that WHO must deliver a timely response when disease outbreaks and emergencies occur. Rokho Kim et al. (819) claim that a Paris climate pact, if ambitious and effective, should also be regarded as a public health treaty to protect vulnerable populations. In the news section, Andr\u00e9ia Azevedo Soares and Menelaos Tzafalias report on how European countries are struggling to respond to the health needs of large numbers of refugees and migrants (822\u2013823). David Nabarro talks to Fiona Fleck about WHO\u2019s transformation into an agency that is fully mandated and equipped to respond to humanitarian emergencies (824\u2013825). Reaching children with congenital heart diseaseSandra da Silva Mattos et al. (881\u2013887) discuss the use of telemedicine networks for remote paediatric cardiology services. Chris Smith et al. (842\u2013850) examine the effect of a mobile-phone based intervention on contraceptive use. Weibing Wang et al. (826\u2013833) estimate excess mortality in a cohort of people with tuberculosis. Ryan K McBain et al. (834\u2013841) investigate the benefits to caregivers of a psychotherapeutic intervention for young people affected by war. Theodoor Visser et al. (862\u2013866) discuss trends and developments in diagnostic devices. Emily Savell et al. (851\u2013861) document exposure to tobacco advertising in 16 countries. Migrants\u2019 health needsLara Miramontes et al. (888\u2013889) call for the inclusion of migrant populations in health impact assessments. Environmental impacts of tobacco Thomas E Novotny et al. (877\u2013880) highlight the environmental impacts of tobacco production and use. Steven J Hoffman et al. (867\u2013876) examine ways to achieve global collective action on antimicrobial resistance."} +{"text": "In the editorial section, Hildy Fong & Eva Harris (438) encourage the fair distribution of health technology. Arthur Chioro et al. (439) discuss future action for managing antimicrobial resistance.In the news section, Gary Humphreys and Fiona Fleck report on the use of the media in public health education (442\u2013443). Fiona Fleck interviews Pali Lehohla on his involvement in the upcoming post-2015 development goals (444\u2013445).Herv\u00e9 Bourhy et al. (503\u2013506) orchestrate a comprehensive disease control training programme.Kathryn Church et al. (457\u2013467) compare national HIV policies in six countries.Xiaolin Wei et al. (407\u2013416) measure patients\u2019 perceptions of primary care in Shanghai and Shenzhen.Andrea B Feigl et al. (468\u2013475) evaluate the effectiveness of a high-school ban on smoking.Serena P Koenig et al. (498\u2013502) investigate a rise in tuberculosis cases following the 2010 earthquake.Susmita Chatterjee & Richard A Gosselin (476\u2013482) compare surgical interventions using two disability weight databases.Kendra N Williams et al. (507\u2013508) call for new questions in household surveys on cooking and fuel collection.Thilde Rheinl\u00e4nder et al. (509\u2013510) argue for the inclusion of shared facilities in development goal measures.Helen S Cox et al. (491\u2013497) describe the need for new treatment regimens.Jose Ma M Angeles et al. (511\u2013512) outline the need for surveillance of the animal reservoir.Daniel J Hruschka et al. (483\u2013490) test a new metric for global wealth estimates.(446\u2013456) investigate the impactof increased food supply onglobal obesity.Stephanie Vandevijvere et al."} +{"text": "Oleg Chestnov et al. (623) explain why noncommunicable disease control is integral to development goals.In editorialsJane Parry (626\u2013627) reports on China\u2019s moves to produce vaccines for the global market. Bruno Vellas (628\u2013629) talks to Fiona Fleck about identifying frail elderly patients in order to improve their quality of life.Christine Leopold et al. (630\u2013640) examine the effects of economic recession on pharmaceutical pricing policies.Mark A Bellis et al. (641\u2013655) examine correlations between harmful behaviour and adverse childhood experiences.Zhongjie Li et al. (656\u2013663) evaluate an early warning system for hand, foot and mouth disease.Ahmed Ehsanur Rahman et al. (664\u2013671) examine a decade of child deaths from diarrhoea.Troy\u00a0D Moon et al. (680\u2013684) describe the use of a mobile clinic for antiretroviral therapy.Aedes aegypti.Rafael Maciel-de-Freitas & Denise Valle (685\u2013689) explain lessons learnt in trying to control Veronica Escamilla et al. (690\u2013694) use satellite imagery to identify households.Susan\u00a0D Cochran et al. (672\u2013679) discuss declassification of disease categories related to sexual orientation.Christine\u00a0G Maure et al. (695\u2013696) introduce the Global Vaccine Safety Initiative."} +{"text": "International statistical classification of diseases and related health problems.In the editorial section, John C Reeder & Winnie Mpanju-Shumbusho (78) argue for a sustained research effort on poverty-related diseases. Emma Allanson et al. (79) announce changes in the way perinatal mortality is categorized by the In the news section, Gary Humphreys (82\u201283) reports on moves to assign adult ratings to films that show actors smoking. W\u00a0Aubrey Webson (84\u201285) tells Fiona Fleck about gains made for people with disabilities.Joseph Adomako et al. (86\u201391) assess the feasibility of community-based maternal mortality surveillance in rural areas.Ana Navas-Acien et al. (92\u2013102) evaluate compliance with anti-smoking legislation.Francisco Rogerl\u00e2ndio Martins-Melo et al. (103\u2013110) estimate mortality from neglected tropical diseases.Jeanne Luh & Jamie Bartram (111\u2013121) correlate access to improved water and sanitation with socioeconomic indicators in 73 countries.Trach Duc Tran et al. (122\u2013129) measure changes in households\u2019 access to iodized salt.Grace J Chan et al. (130\u2013141) review the factors that help and hinder kangaroo care.Barbara McPake et al. (142\u2013146) consider dual public and private sector practice by health workers.Thelma Tupasi et al. (147\u2013152) describe a global capacity-building programme to develop a new treatment for multidrug-resistant tuberculosis.Abdul Momin Kazi & Lubna Ashraf Jafri (153\u2013154) discuss the use of mobile phones in polio eradication.Rob E Dorrington & Debbie Bradshaw (155\u2013156) call for more investment in civil registration and vital statistics."} +{"text": "For exal., 2015). Adjustal., 2015) is the in vitro systems , partical., 2014; Grinberal., 2014; Ghallabal., 2014; Schug eal., 2014), neurotal., 2014; Krug etal., 2014; St\u00f6ber,al., 2014; Faiz etal., 2011; Sterneral., 2011; Strikwoal., 2011; Wang etal., 2011). Furtheal., 2011; Drasdo al., 2011; Widera,al., 2011). The he"} +{"text": "In the editorial section, Flavia Bustreo and Robin Gorna (286) call for papers that link millennium to sustainable development goals on women\u2019s, children\u2019s and adolescents\u2019 health. Lisa\u00a0G Johnston et al. (287) discuss ways to obtain information on HIV status; a topic debated in a round table led by Rachel Baggaley et al. (353\u2013358).In the news section, Gary Humphreys (290\u2013291) reports on a technology transfer collaboration between Argentina and the Netherlands. Fiona Fleck interviews Adetokunbo Lucas (292\u2013293) about the five decades he has spent working in public health.Stefanie Vandevijvere et al. (294\u2013302) study implementation of healthy food policies.Richard Matzopoulos et al. (303\u2013313) collect data on deaths caused by injuries.Reinhard Kaiser et al. (314\u2013319) review vaccination coverage survey reportsMuhammed\u00a0Olanrewaju Afolabi et al. (320\u2013328) test a multimedia consent tool for research participants.Michael Roerecke et al. (329\u2013338) recalculate national estimates of alcohol-related oesophageal cancer.Joanna\u00a0F Crofts et al. (347\u2013352) train hospital staff to confront obstetric emergencies.Mira Johri et al. (339\u2013346) review strategies designed to increase demand for childhood vaccinations.Oladele A Ogunseitan (359\u2013360) documents missed opportunities to prevent asbestos-related diseases."} +{"text": "In the editorial section, Christopher Dye et al. (158) encourage all researchers to share their data on Zika virus as quickly and widely as possible. Michelle J Hindin et al. (159) argue that pregnancies as well as birth rates should be tracked as development indicators. Yoko Akachi et al. (160) call for papers measuring quality of clinical care in low- and middle-income countries.In the news section, Carolyn Mahoney and Fiona Fleck report on how surgical provision falls far short of what is needed in developing countries (163\u2013164). Ana Bispo tells Andr\u00e9ia Azevedo Soares how scientists are scrambling to develop better tests for Zika infections (165\u2013166). Paulina Correa-Burrows et al. (185\u2013192) explore the associations between a healthy diet and academic performance among 16 year-old students.Supporting elderly people with dementia Christina Wu et al. (167\u2013173) evaluate health care and support in a rural area. HIV-related services online Weibin Cheng et al. (222\u2013227) describe a community-based project to expand access to testing and treatment services for HIV. Catastrophic medical bills Stephen Jan et al. (193\u2013200) examine the out-of-pocket costs of hospitalization for acute coronary syndromes.Surviving Ebola Tine Van Bortel et al. (210\u2013214) study the psychosocial impacts of the epidemic on affected individuals and communities. Sebastien Antoni et al. (174\u2013184) compare GLOBOCAN estimates with national data. Thomas G Weiser et al. (201\u2013209) estimate global surgical volumes and gaps in 2012.Adeladza Kofi Amegah & Jouni JK Jaakkola (215\u2013221) recommend actions for improving indoor air quality.Andrew D Jones & Gebisa Ejeta (228\u2013229) propose changes to the way food crops are produced. Nicola C Richards et al. (230\u2013232) argue that more data are needed on disabilities as outcomes of non-communicable diseases."} +{"text": "Het doel van dit onderzoek was het evalueren van de ervaren kwaliteit van het leerklimaat door artsen in opleiding (aios) binnen de medische vervolgopleiding arts Maatschappij\u202f+\u2009Gezondheid (M\u202f+\u2009G) en het vergelijken van de percepties van aios en supervisors.Aios van vijf profielen arts M\u202f+\u2009G, die in 2019 en later met hun opleiding zijn begonnen, en supervisors die betrokken zijn bij het opleidingsprogramma werden uitgenodigd om een online vragenlijst in te vullen op basis van een aangepaste versie van de D\u2011RECT-vragenlijst. De antwoorden van aios en supervisors van dezelfde opleidingsinstelling en hetzelfde profiel werden gematcht om de waargenomen kwaliteit van het leerklimaat te vergelijken.Honderdveertien aios reageerden . De algemene beoordeling van het leerklimaat door de aios gaf een gemiddelde score van 4,19 op een vijfpuntsschaal. Achtendertig supervisor-aios-matches werden gevormd. Er waren geen noemenswaardige verschillen in de beleving van aios en supervisors.De algemene beoordeling van het leerklimaat door de aios was positief. De percepties van het leerklimaat door supervisors en aios zijn vergelijkbaar. Onze aangepaste versie van D\u2011RECT lijkt geschikt om het leerklimaat van de medische vervolgopleiding arts M\u202f+\u2009G te evalueren. Herhaling van het onderzoek is nodig om het leerklimaat op lokaal niveau te beoordelen en onze bevindingen te bevestigen. Verdere aanpassing en validering van de vragenlijst is wenselijk voor een betere weergave van de werk- en leeromgeving van de arts M\u202f+\u2009G.De online versie van dit artikel (10.1007/s12508-022-00367-6) bevat aanvullend materiaal, toegankelijk voor daartoe geautoriseerde gebruikers. Leerklimaat is een sleutelcomponent van de kwaliteit van de medische vervolgopleiding en kan worden gedefinieerd als de sociale, emotionele en fysieke omstandigheden waaronder de arts in opleiding (aios) kennis verwerft \u20133. Het oBij het evalueren van het leerklimaat kan het van toegevoegde waarde zijn om ook het perspectief van de supervisors mee te nemen. Uit eerder onderzoek blijkt dat werken in dezelfde omgeving, maar met een andere rol, kan leiden tot een andere beoordeling van het leerklimaat. Medische studenten en chirurgische arts-assistenten waren het eens over universele elementen die bijdragen aan een optimaal leerklimaat voor alle leerlingen, maar verschilden van mening over elementen die meer verband houden met de verschillen in hun opleidingsniveau . PerceptIn 2019 is het curriculum voor de aios in opleiding tot arts Maatschappij\u202f+\u2009Gezondheid (M\u202f+\u2009G) ingrijpend gewijzigd. Deze aios zijn nu in dienst bij \u00e9\u00e9n landelijke werkgever, waardoor het mogelijk is om de praktijkleerperiodes en stages bij verschillende opleidingsinstellingen te volgen. Voorheen volgden de aios hun hele opleiding op de werkplek waar ze voor aanvang van hun opleiding al werkzaam waren. Zo\u2019n grote curriculumverandering kan impact hebben op het leerklimaat.Het werkveld van de arts M\u202f+\u2009G is onderverdeeld in profielen . De NethDe aios volgen een praktijkgerichte opleiding ondersteund door cursorisch onderwijs, dat door de opleidingsinstituten op centrale locaties verzorgd wordt. Op fulltimebasis behelst de praktijkopleiding twee periodes van in totaal vijftien maanden praktijkleren in het eigen profiel, aangevuld door drie keer drie maanden externe stages. Werkplekleren in het eigen profiel vindt plaats bij opleidingsinstellingen door heel Nederland, zoals GGD\u2019en, het Rijksinstituut voor Volksgezondheid en Milieu (RIVM) en Centra voor Jeugd en Gezin. Tijdens een praktijkperiode wordt iedere aios gekoppeld aan een praktijkopleider op de werkvloer. De praktijkopleider fungeert als mentor en bewaakt de voortgang van de praktijkopleiding. Daarnaast zijn andere artsen werkzaam in dezelfde instelling vaak betrokken bij het superviseren en begeleiden van aios bij werkplekleren. Instituutopleiders verbonden aan de opleidingsinstituten NSPOH en TNO bewaken de voortgang van de aios en geven vorm aan de opleiding, rekening houdend met de landelijke opleidingskaders.De NSPOH evalueert het curriculum volgens een cyclisch proces zoals gedefinieerd door de kwaliteitsrichtlijnen van de Koepel Artsen Maatschappij\u202f+\u2009Gezondheid . Tot nu Het doel van dit onderzoek is daarom om de ervaren kwaliteit van het leerklimaat door aios M\u202f+\u2009G binnen de opleidingsinstelling op nationaal niveau per profiel te evalueren. Ons tweede doel is om het perspectief ten aanzien van het leerklimaat van de aios en supervisors met elkaar te vergelijken. Dit onderzoek richt zich op werkplekleren in het kader van het eigen profiel, het leerproces tijdens de externe stages blijft buiten beschouwing. Voor zover bij ons bekend, is ons onderzoek het eerste naar het leerklimaat binnen de arts M\u202f+\u2009G-profielopleiding vanuit het perspectief van de aios en supervisor.In juni 2021 zijn alle 224 Nederlandse aios M\u202f+\u2009G die in 2019 en later met hun opleiding zijn begonnen, en hun 195 praktijkopleiders gevraagd om deel te nemen aan ons onderzoek. Aios en praktijkopleiders zijn direct per e\u2011mail uitgenodigd, gebruikmakend van de inschrijfgegevens van de aios en praktijkopleiders van de NSPOH en TNO. Aios werd gevraagd om hun huidige praktijkperiode te evalueren. Wanneer de aios ten tijde van het onderzoek een externe stage liep, werd deze verzocht om de vragen voor de meest recente praktijkperiode van het eigen profiel in te vullen. Verder hebben we de contactpersonen van de 39\u00a0praktijkopleidingsinstellingen gevraagd om onze vragenlijst door te sturen naar alle andere artsen binnen de praktijkopleidingsinstelling die betrokken zijn bij het opleiden van aios. De reacties van artsen van die laatsten en die van de praktijkopleiders werden gegroepeerd, omdat ze allemaal het perspectief van de begeleider vertegenwoordigen. Beide groepen worden onder de term supervisor geschaard.De vragenlijsten werden ingevuld via een online platform (SurveyMonkey). Tot drie keer toe is er een automatische herinnering per e\u2011mail verstuurd naar non-responders. De gegevensverzameling duurde een maand.Als ik een supervisor nodig heb, kan ik er altijd een bereiken werd veranderd in een supervisorvraag Als een aios een supervisor nodig heeft, kan hij/zij er altijd een bereiken.We gebruikten de Dutch Residency Educational Climate Test (D-RECT) als basis om ons onderzoek uit te voeren , 24. De Met beschrijvende statistieken werden de kenmerken van de onderzoekspopulatie samengevat. Statistische analyses werden uitgevoerd in SPSS-28.Ten tweede werd de interne consistentie van de subschalen met behulp van Cronbach\u2019s\u00a0\u03b1 geschat, zowel voor de vragenlijst van de aios als voor die van de supervisor. In het algemeen wordt een \u03b1\u2011waarde van 0,70 of meer als bevredigend beschouwd. Aangepaste items werden getest op hun bijdrage aan Cronbach\u2019s\u00a0\u03b1. Als Cronbach\u2019s\u00a0\u03b1 met 0,02 of meer verbeterde door het aangepaste item te verwijderen, hebben we dit item niet opgenomen in de subschaal en afzonderlijk gerapporteerd.Ten derde werden de gemiddelde uitkomstscores berekend voor elke D\u2011RECT-subschaal door de totale subschaalscore te delen door het aantal subschaalvragen. De totale gemiddelde D\u2011RECT-score werd berekend als het gemiddelde van alle itemscores gedeeld door het totale aantal vragen. Gemiddelde scores werden voor aios en supervisors afzonderlijk berekend en opgesplitst per profiel. Scores boven de 4\u00a0duiden op een positieve houding ten opzichte van de beoordeelde subschaal.Ten slotte werden de antwoorden van aios en supervisors per profiel op het niveau van opleidingsinstelling gematcht om de beleving van aios en supervisors te kunnen vergelijken. Subschaalgemiddelden en itemscores van de aios werden afgetrokken van de subschaalgemiddelden van hun gematchte supervisor. Het gemiddelde van het verschil tussen aios en supervisors werd berekend. Reacties van aios en supervisors die niet konden worden gematcht, zijn buiten deze analyse gelaten. Subschaalverschillen van\u00a00,6 of hoger werden als opmerkelijk beschouwd.Ethische goedkeuring is ontvangen van de NVMO-Ethische Toetsingscommissie, protocolnummer 2021.3.4. De deelnemers werd verteld dat anonimiteit gegarandeerd was en dat deelname vrijwillig was. Informed consent werd verkregen.In totaal hebben 114 van de 224 aios M\u202f+\u2009G die in 2019 en later met hun opleiding zijn begonnen gereageerd . Van de JGZ- aios was 93\u202f% vrouwelijk, voor het ITM- en FG-profiel was dit respectievelijk 78,1\u202f% en 54,6\u202f%.Cronbach\u2019s\u00a0\u03b1-test resulteerde in het verwijderen van \u00e9\u00e9n toegevoegde vraag uit de D\u2011RECT-subschaal Toegankelijkheid supervisors (vraag\u00a05) en \u00e9\u00e9n toegevoegde vraag uit de subschaal Begeleiden en toetsen (vraag\u00a011).De uitkomstscores zijn samengevat in tab.\u00a0In totaal ontvingen we reacties van 106 supervisors van 36\u00a0opleidingsinstellingen. Voor 78,8\u202f% 26/33) van de opleidingsinstellingen die het profiel JGZ aanbieden hebben we een match kunnen vormen tussen aios en supervisor. Een match werd gevormd voor 38,1\u202f% (8/21) van de opleidingsinstellingen die het ITM-profiel aanbieden en voor 33,3\u202f% (5/15) die het FG-profiel aanbieden. De gemiddelde verschillen tussen supervisors en aios waren over het algemeen minder dan\u00a00,6, waarbij positieve scores erop wezen dat de supervisors de subschaal iets hoger beoordeelden dan de aios (tab.\u00a06/33 van De algemene beoordeling van het leerklimaat door aios M\u202f+\u2009G was positief, met een gemiddelde score van 4,19 op een vijfpuntsschaal. Samenwerking tussen aios oftewel Samenwerking peers scoorde het minst gunstig, vanwege de lage scores op samenwerking tussen profielen. Er waren geen noemenswaardige verschillen in de beleving van aios en supervisors met betrekking tot het leerklimaat.Nederlandse aios M\u202f+\u2009G ervaren het leerklimaat op landelijk niveau als positief. De lagere totale gemiddelde score en subschaalscores voor het profiel FG, vergeleken met de scores van alle aios M\u202f+\u2009G, moeten voorzichtig worden ge\u00efnterpreteerd vanwege het kleine aantal aios profiel FG en de relatief lagere respons in dit profiel. Een bijkomend aspect is dat de FG-opleiding een grotere verandering heeft ondergaan dan de andere profielen en volledig is vernieuwd van een cursus van enkele maanden naar een tweejarige opleiding. Het is aannemelijk dat de FG-opleidingsinstellingen door deze curriculumverandering voor een grotere uitdaging hebben gestaan dan de andere profielen, waar de curriculumveranderingen minder ingrijpend waren.M\u202f+\u2009G-scores voor de subschaal Samenwerking peers waren slecht in vergelijking met de andere subschalen. Dit bleek het gevolg van slechte scores voor het aspect samenwerking tussen aios van verschillende profielen, terwijl de samenwerking binnen het profiel beter werd beoordeeld. Om de aios M\u202f+\u2009G voor te bereiden op de praktijk van de eenentwintigste eeuw is in het vernieuwde curriculum een van de doelen het stimuleren van samenwerking tussen alle profielen arts M\u202f+\u2009G . MaatschVolgens onze bevindingen lijken er geen grote verschillen te zijn in de beleving van het leerklimaat tussen aios M\u202f+\u2009G en hun supervisors. Voor een aantal subschalen in de profielen ITM en FG werden echter wel verschillen gevonden tussen de beleving van de aios en die van de supervisor, maar vanwege het kleine aantal aios-supervisor-matches voor deze profielen kunnen hieruit geen harde conclusies worden getrokken.Onderzoek onder aios chirurgie en hun supervisors naar aspecten die onderdeel zijn van het leerklimaat, zoals onderwijsprestaties en feedback , 27, lieVoor zover ons bekend is dit het eerste onderzoek dat het leerklimaat in de medische vervolgopleiding arts M\u202f+\u2009G evalueert en daarbij de percepties van aios en supervisors van het leerklimaat van de aios M\u202f+\u2009G vergelijkt. Een kracht van ons onderzoek is de landelijke opzet, waarbij alle aios M\u202f+\u2009G en hun supervisors zijn uitgenodigd om deel te nemen aan het onderzoek. Een ander sterk punt is ons behoorlijke responspercentage van ongeveer 50\u202f% voor aios en een match tussen aios en supervisor voor 78\u202f% van de opleidingsinstellingen voor het JGZ-profiel. Het feit dat we ondanks aanpassingen aan de originele gevalideerde D\u2011RECT-vragenlijst betrouwbare Cronbach\u2019s\u00a0\u03b1-waarden hebben gevonden, versterkt de waarde van onze resultaten.Een beperking van dit onderzoek was het aantal aios en supervisors ITM en FG dat per opleidingsinstelling beschikbaar was. Daarnaast zijn door het beperkte aantal aios in opleiding tot arts M\u202f+\u2009G, niet alle supervisors van iedere opleidingsperiode aan een aios gekoppeld. Daardoor was het percentage supervisor-aios-matches voor ITM- en FG-profielen laag, en konden we voor de meeste opleidingsinstellingen het geadviseerde minimum van drie aios of supervisors niet bereiken. Dit kan van invloed zijn geweest op de betrouwbaarheid van de gevonden verschillen tussen aios en supervisors. Het ontbreken van verschillen in perceptie kan ook worden veroorzaakt door het gebruikte instrument. Een vijfpuntslikertschaal geeft een beperkte mogelijkheid om onderscheid te maken tussen niveaus van overeenstemming. Dit maakt het moeilijker om kleine verschillen in perceptie van het leerklimaat te detecteren .Dit onderzoek laat zien dat de D\u2011RECT-vragenlijst na kleine aanpassingen geschikt is voor het beoordelen van het leerklimaat van de medische vervolgopleiding arts M\u202f+\u2009G, met betrouwbare Cronbach\u2019s\u00a0\u03b1-waarden voor zowel de vragenlijst voor aios als die voor supervisors. Vanwege de kleine aantallen per opleidingsinstelling moeten de uitkomsten worden gezien als een eerste indicatie van de perceptie van het leerklimaat.Het onderzoeken van gepercipieerde verschillen tussen aios en supervisors is relevant, omdat het richting geeft aan het verbeteren van het leerklimaat. Bij duidelijke verschillen in beleving moeten de supervisors om te beginnen inzicht krijgen in de behoeften van de aios en is het interessant om te achterhalen waarom supervisors het leerklimaat anders ervaren. Wanneer aios en supervisors het leerklimaat gelijkelijk beoordelen, kan de aandacht direct worden gericht op de verbeterpunten. In dit artikel blijkt dat het leerklimaat binnen de opleiding arts M\u202f+\u2009G landelijk als positief wordt ervaren, maar ook dat er wel lokale verschillen kunnen zijn die aandacht behoeven.Om aandachtspunten met betrekking tot werkplaatsleren in de publieke gezondheid in meer detail te onderzoeken, is het aan te raden om alle relevante actoren erbij te betrekken. Hiervoor zijn naast de aios en de supervisors de volgende actoren van belang: instituutopleiders, directeuren van de opleidingsinstellingen, lokale teamleiders en opleidingsmanagers verantwoordelijk voor de implementatie van de opleiding. De aios, supervisors en instituutopleiders worden door middel van kwalitatieve interviews verdiepend gevraagd naar verbeterpunten van de nieuwe opleiding. De directeuren en managers zijn behalve voor de bedrijfsvoering ook verantwoordelijk voor het implementeren van de opleiding op lokaal niveau. Om de samenwerking tussen de verschillende profielen te bevorderen zijn onderwijskundige middelen zoals profieloverstijgende opdrachten nodig. Daarnaast is de ondersteuning op bedrijfsvoeringsniveau voor profieloverstijgend samenwerken een cruciale factor. De invloed van de bedrijfsvoering en mogelijk conflicterende bestuurskundige belangen op de opleiding arts M\u202f+\u2009G is onvoldoende bekend. GGD-directeuren en -managers worden betrokken bij vervolgonderzoek naar deze aspecten.De vragenlijst kan verder aangepast worden aan de specifieke werk- en leeromgeving van de arts M\u202f+\u2009G. Andere kwaliteitsdomeinen, zoals beschreven in de kwaliteitsrichtlijnen van de Koepel Artsen Maatschappij\u202f+\u2009Gezondheid , kunnen Ons onderzoek wijst op een positief leerklimaat voor aios M\u202f+\u2009G. Omdat het onderwijs altijd evolueert en verandert, is toekomstig onderzoek nodig om het leerklimaat in de loop van de tijd te blijven evalueren.Herhaalde evaluaties zijn ook nodig om voldoende gegevens te verzamelen om conclusies te trekken over het leerklimaat op het niveau van opleidingsinstellingen, vooral voor de kleinere profielen ITM en FG. Om de respons te verhogen wordt voor de aios een link naar de anonieme vragenlijst aan het portfolio toegevoegd. Supervisors worden jaarlijks via e\u2011mail benaderd. Het belang van het invullen van de vragenlijst voor de tweejaarlijkse KOERS-kwaliteitscyclus van de opleidingsinstelling wordt in de correspondentie benadrukt.Aios M\u202f+\u2009G beoordelen het leerklimaat van de opleidingsinstelling als positief en aios en supervisors lijken het leerklimaat niet anders te ervaren. Samenwerking van aios tussen profielen behoeft nader onderzoek voordat duidelijk is of verbetering nodig is en hoe werkplaatsleren daarin ondersteuning kan bieden. Onze aangepaste versie van de D\u2011RECT-vragenlijst lijkt geschikt om het leerklimaat binnen de medische vervolgopleiding arts M\u202f+\u2009G te evalueren, maar vervolgonderzoek is nodig om onze vragenlijst te valideren en onze bevindingen te bevestigen."} +{"text": "In the editorial section, Etienne V. Langlois et al. (2) argue for better postnatal care services. Karen I. Barnes et al. (3) describe the contributions made to clinical pharmacology by Peter Ian Folb.Tatum Anderson (6\u20137) reports on emergency response initiatives in Botswana, Mauritania, Niger, Nigeria and Togo. Anthony Fauci talks to Gary Humphreys (8\u20139) about his work with the National Institute of Allergy and Infectious Diseases. Merlin L. Willcox et al. (62\u201375) review characteristics of mortality surveillance.Sonam Vijay et al. (20\u201327) track stewardship in tertiary hospitals.Rajaram Subramanian Potty et al. (28\u201335) assess support group participation.Parsa Erfani et al. (10\u201319) compare costs of biomarker analysis methods.Loai Albarqouni et al. (36\u201361) quantify the extent of unnecessary treatments."} +{"text": "In the editorial section, Hanna Ter\u00e4s & Nellie Kartoglu (162) explain how civil society organizations contribute to emergency preparedness. Edgar Manuel Cambaza (163) proposes ways to expand the provision of nurses\u2019 education.Tatum Anderson (166\u2013167) reports on efforts to improve diagnostics and treatments for invasive fungal pathogens. Rita Oladele talks to Gary Humphreys (168\u2013169) about the need for more investment in invasive fungal pathogen surveillance, research and clinical capacity.Jacobo Antona-Makoshi et al. (211\u2013222) estimate potential death and disability averted.Jean Patrick Ouamba et al. (170\u2013178) target prevention measures.Oliver Huse et al. (226\u2013228) introduce a landscape analysis of factors contributing to obesity.Aakshi Kalra et al. (179\u2013190) track case notifications.Rakhi Dandona et al. (191\u2013201) compare two sources of data.Eva Nabawanuka et al. (202\u2013210) examine chest radiographs of hospitalised children.Juan J Carrique-Mas et al. (223\u2013225) describe new legislation and challenges to full compliance."} +{"text": "In editorials this month, Kapil Narain et al. (582) discuss strategies for malaria vaccination during the COVID-19 pandemic in African countries. D\u00e9vora Kestel et al. (583) present a WHO report on the transformation needed in mental health care.Vijay Shankar Balakrishnan and Gary Humphreys (586\u2013587) report on the ways in which the monkeypox epidemic is highlighting the risks entailed in neglecting local outbreaks. Vera Cordeiro talks to Andr\u00e9ia Azevedo Soares (588\u2013589) about creating a community-level strategy to tackle the interlinked conditions of poverty and disease.Pascal Geldsetzer et al. (601\u2013609) analyse survey data from demographic surveillance sites.Tigest Tamrat et al. (590\u2013600) transform paper registers into a digital system.Fernando Korn Malerbi & Gustavo Barreto Melo (643\u2013647) test the feasibility of using artificial intelligence.Aashna Mehta et al. (610\u2013619) track sales data.Zila M Sanchez et al. (628\u2013635) describe the process of developing legislation.Geofrey Makenga et al. (651\u2013652) make the case for regional capacity development.Juleen Lam et al. (620\u2013627) model global economic costs. Iris R Joosse et al. (636\u2013642) delineate data gaps. Joseph Friedman et al. (648\u2013650) define a research agenda."} +{"text": "In editorials this month, Stefan Listl et al. (294) call for harnessing citizens\u2019 values through deliberative processes to improve oral health systems. Samira Choudhury et al. (295) discuss how lessons from the COVID-19 pandemic create opportunities to manage antimicrobial resistance.Lynn Eaton (298\u2013299) reports on antimicrobial misuse during the COVID-19 pandemic and the lack of adherence to treatment protocols. Parameswaran Iyer talks to Gary Humphreys (300\u2013301) about how to transform water, sanitation and hygiene practices through behavioural change interventions.King et al. (302\u2013314) identify prognostic associations with hypoxaemia or hypoglycaemia.Ning et al. (329\u2013336) measure helmet use by cyclists and motorcyclists. Eze et al. (337\u2013351) estimate incidence and trends of catastrophic expenditure.Opiyo et al. (352\u2013354) present a global data-sharing platform for facility-level rates. Yoo et al. (315\u2013328) track COVID-19 vaccine allocation and distribution through COVAX."} +{"text": "In the editorial section Shannon Doocy et al. (466) call for a continuing crisis response in Venezuela. Claudia Chamas et al. (467) summarize recent initiatives to address diagnostic technology gaps in low- and middle-income countries.Tatum Anderson (470\u2013471) describes WHO\u2019s work to improve reporting of attacks on health care in conflict settings. Ahmed Hankir talks to Vijay Shankar Balakrishnan (472\u2013473) about his efforts to combat stigma surrounding mental health. Ianka Cristina Celuppi et al. (520\u2013524) introduce a database used to deploy health workers nationwide. Himanshu Nayak et al. (484\u2013490) implement a screening programme in Ahmedabad.Ali Hosseinzadeh et al. (474\u2013483) compare cases, hospitalizations and deaths.Khalid Almoteiry et al. (511\u2013519) detail evolution and features of a national policy.Christopher P Seaman et al. (491\u2013502) review the evidence for the controlled temperature chain approach.Anne Ammerdorffer et al. (503\u2013510) examine one way to improve access."} +{"text": "According to Lopez et al. pancreatic stone protein (PSP) is \u201ca promising biomarker to diagnose infections in hospitalized patients,\u201d distinguishing infections from non-infections and outperforming C-reactive protein (CRP) and procalcitonin (PCT) . PSP is"} +{"text": "Dit bevestigt de rationale achter het eerdere beleid ten aanzien van het langer gesloten houden van binnensportaccommodaties om het aantal besmettingen te reduceren.Binnensportaccommodaties werden langer gesloten gehouden dan buitensportaccommodaties gedurende de lockdowns tijdens de COVID-19-pandemie. Dit onderzoek beantwoordt de vraag of binnen sporten het risico op besmetting met SARS-CoV\u20112 vergroot. Hiervoor werden gegevens gebruikt van de COVID RADAR-app. Er werd gecorrigeerd voor leeftijd, vaccinatiestatus, geslacht, ander risicogedrag, prevalentie van SARS-CoV\u20112 en kwaliteit van de leefomgeving. Uit analyses van 1.353 gebruikers, van wie 13,0% een positieve testuitslag rapporteerde, blijkt dat binnensporters significant vaker een positieve test hadden bevat aanvullend materiaal, toegankelijk voor daartoe geautoriseerde gebruikers. Na de lockdowns gedurende de COVID-19 (COronaVIrus Disease 2019)-pandemie in Nederland werden buitensportaccommodaties eerder geopend dan binnensportaccommodaties. Het idee hierachter is ook nu nog dat de kans op een besmetting met SARS-CoV\u20112 (severe acute respiratory syndrome coronavirus\u00a02) binnen in een zaal of een sportschool groter is dan in buitensportaccommodaties. Bewijs daarvoor ontbreekt tot nu toe [De COVID RADAR-app is een gratis smartphone-app waarin vragen gesteld worden over symptomen en gedrag (waaronder het houden van afstand en sportgedrag). Ook kunnen gebruikers aangeven dat ze onlangs getest zijn op SARS-CoV\u20112 en wat de uitslag van die test was. In een eerdere publicatie hebben we beschreven dat de COVID RADAR-app een valide meetinstrument is voor surveillance van SARS-CoV\u20112, waarbij incidentie van symptomen toeneemt rond een positieve test, en dat verplaatsings- en contactgedrag in de weken voor een positieve test bovengemiddeld hoog waren [Dit verslag beantwoordt met behulp van data van de COVID RADAR-app de vraag of mensen die binnen sporten daadwerkelijk vaker besmetting met SARS-CoV\u20112 rapporteren dan mensen die buiten sporten.Voor deze analyse werden data gebruikt van de COVID RADAR-app . Meer daOmdat inzichten gedurende de tijd veranderden is in de bijna twee jaar dat de app in de lucht was, de inhoud enkele malen aangepast. Zo is na een half jaar de vraag over sporten toegevoegd (augustus 2020), kwam er in januari 2021 de mogelijkheid bij om een negatieve test te rapporteren en konden gebruikers aangeven of ze gevaccineerd waren. Gezien deze aanpassingen (maar ook vanwege veranderingen in het testbeleid) zijn in de huidige analyses alleen gerapporteerde testuitslagen van na 1\u00a0januari 2021 ge\u00efncludeerd.Het theoretische beloop tussen besmetting en rapportage van een testuitslag in de app is op te delen in verschillende fasen. Ten eerste vertoont een individu risicogedrag, wat leidt tot virale transmissie. Vervolgens ontwikkelen zich na de incubatieperiode (gemiddeld 2\u201314\u00a0dagen) symptomen bij het ge\u00efnfecteerde individu. Deze maakt dan een afspraak voor een test op de aanwezigheid van SARS-CoV\u20112 (1\u20133\u00a0dagen). Vervolgens wordt de test afgenomen en wordt de testuitslag bekend, die uiteindelijk in de app gerapporteerd wordt (2\u20133\u00a0dagen). Omdat het precieze moment van besmetting met SARS-CoV\u20112 nooit met zekerheid vast te stellen is, gebruiken we een tijdsperiode van 10\u201320\u00a0dagen voor een testuitslag waarin het risicogedrag en dus ook de besmetting mogelijk heeft plaatsgevonden. Deze theoretische tijdsperiode wordt bevestigd in onze eerdere publicatie, waarin gemiddeld gedrag van gebruikers die uiteindelijk positief testen, hoger was in de tijdsperiode van 10\u201320\u00a0dagen voor een positieve test [In de app konden gebruikers aangeven of ze onlangs getest waren en wat de uitslag van de test was. Voor deze analyse includeren we per gebruiker alleen eerste vermeldingen van een positieve of negatieve test. Personen die zowel een positieve als negatieve testuitslag rapporteerden konden tweemaal worden meegenomen in de analyse, onder voorwaarde dat deze tests minimaal 60\u00a0dagen na elkaar werden gerapporteerd.In de app werd ook gevraagd of de gebruiker aan sport deed. De antwoordmogelijkheden waren: nee, ja (bij een vereniging of sportschool binnen), ja (bij een vereniging of sportschool buiten) of ja . Voor het beantwoorden van de onderzoeksvraag selecteerden we alleen gebruikers die binnen de periode van mogelijke besmetting minimaal \u00e9\u00e9n keer hadden gesport, zowel binnen als buiten. Een gebruiker sportte binnen als er werd geantwoord: \u2018ja, bij een vereniging of sportschool binnen\u2019. Omdat personen die hadden geantwoord: \u2018ja \u2019 zowel binnen als buiten konden sporten, werden zij ge\u00ebxcludeerd.Omdat vaccinatiestatus de kans op het ge\u00efnfecteerd raken be\u00efnvloedt, werd hiervoor gecorrigeerd. Een gebruiker is gevaccineerd als deze bij alle observaties in de mogelijke besmettingsperiode aangeeft volledig gevaccineerd te zijn. Leeftijdscategorie\u00ebn werden gecategoriseerd in <\u202f18\u00a0jaar, 19\u201349\u00a0jaar, 50\u201369\u00a0jaar en \u2265\u202f70\u00a0jaar. Gender werd gecategoriseerd in man\u00a0(1) en niet man\u00a0(0), inclusief non-binaire genders.Omdat sociale en regionale omgevingsfactoren de kans op besmetting met SARS-CoV\u20112 be\u00efnvloeden hebben wij hiervoor gecorrigeerd . DaarvooAndere mogelijke factoren die de kans op besmetting met SARS-CoV\u20112 be\u00efnvloeden zijn andere vormen van risicogedrag en de periodeprevalentie van SARS-CoV\u20112. Hiervoor hebben we gecorrigeerd door het gemiddelde van de antwoorden van de gebruiker op de vraag over het aantal personen binnen de 1,5\u202fmeter in het geselecteerde tijdvak van 10\u201320\u00a0dagen voor een positieve test te gebruiken. Om te corrigeren voor de SARS-CoV-2-prevalentie hebben we openbare data van het Rijksinstituut voor Volksgezondheid en Milieu (RIVM) gebruikt over het aantal dagelijks positieve SARS-CoV-2-tests per gemeente . Om de pbinomial generalized linear mixed model gebruikt, met als uitkomstvariabele een positieve SARS-CoV-2-test, om verder het verband met sporten te onderzoeken. Om te corrigeren voor clustering als gevolg van herhaalde metingen, werd er een random intercept toegevoegd op individueel niveau. Effecten zijn gerapporteerd in de vorm van een oddsratio (OR) en robuuste 95%-betrouwbaarheidsintervallen (BI). Er werd gecorrigeerd voor leeftijd, vaccinatiestatus, gender, gemiddeld aantal personen binnen de 1,5\u202fmeter gerapporteerd, periodeprevalentie van SARS-CoV\u20112 en mate van kwaliteit van de leefomgeving van de gebruiker.Vervolgens werd er een In totaal werden drie analyses uitgevoerd: een hoofdanalyse naar het verband tussen minimaal \u00e9\u00e9n keer binnen sporten en een positieve test, en twee subanalyses. Omdat in de hoofdanalyse binnensporters ook buiten konden sporten werd in de eerste subanalyse dezelfde analyse uitgevoerd als bij de hoofdanalyse, maar in een andere gebruikersgroep, namelijk gebruikers die of alleen maar binnen sporten of alleen maar buiten sporten. De tweede subanalyse werd in dezelfde gebruikersgroep als de hoofdanalyse uitgevoerd, maar nu werd het verband tussen iedere dag extra binnen sporten en een positieve test getest. Analyses werden uitgevoerd in STATA\u00a016.1 .n\u202f=\u20091.353) sportte 43% ooit binnen. Dit is ook vergelijkbaar met Nederlandse statistieken [p\u202f<\u20090,001) en hadden vaker de app ingevuld . De kwaliteit van de leefomgeving was significant lager bij binnensporters . Het gemiddelde aantal personen binnen de 1,5\u202fmeter was hoger bij binnensporters . De gemiddelde prevalentie van SARS-CoV\u20112 ten tijde van het sporten was bij binnensporters hoger .Ongeveer 50% van de COVID RADAR-app-gebruikers sportte minimaal eenmaal per week, wat overeenkomt met het gemiddelde in Nederland . In totaistieken . Van henp\u202f=\u20090,003)).Van de gebruikers die alleen buiten hadden gesport was 8,3% van de testuitslagen positief. Van de gebruikers die minimaal \u00e9\u00e9n keer binnen hadden gesport was 19,4% van de testuitslagen positief zie tab.\u00a0. Na corrp\u202f=\u20090,003) . Na correctie bleek een OR van 1,84 . Wanneer we specifiek kijken naar COVID RADAR-gebruikers die alleen maar binnen hadden gesport was er geen significant verschil met de hoofdanalyse. Er kon na correctie geen significant verband worden aangetoond tussen het aantal dagen binnen sporten en de testuitslag.We vonden in de met de COVID RADAR-app verzamelde gegevens dat het binnen sporten samenhangt met een hogere kans op een positieve SARS-CoV-2-testuitslag dan het buiten sporten voor een testuitslag te kiezen.Ook de subanalyse en overige correcties die we hebben uitgevoerd vormen sterke kanten van deze analyse en wijzen in de richting van een oorzakelijk verband. Het was niet mogelijk om te corrigeren voor andere confounders (bijvoorbeeld comorbiditeit) in de relatie tussen een positieve test en het wel of niet buiten sporten. Daarom kunnen geen uitspraken gedaan worden over de impact die deze hogere kans op besmetting met SARS-CoV\u20112 op de zorgvraag zou kunnen hebben. Ook waren er geen gegevens bekend over het soort sport dat werd beoefend. Mogelijk is er een verschil tussen teamsporters en individuele sporters wat betreft de kans op besmetting met SARS-CoV\u20112.Uit de analyse van data die met de COVID RADAR-app werden verzameld, blijkt dat wanneer iemand sport, binnen sporten samenhangt met een hogere kans op een positieve testuitslag ten opzichte van het buiten sporten. Dit bevestigt de rationale achter het eerdere beleid ten aanzien van het langer gesloten houden van binnensportaccommodaties om het aantal besmettingen te reduceren."} +{"text": "Thoughal., 2016). As theal., 2016; Gu et aal., 2016; Hao et al., 2016; Kim et al., 2016; Lei et al., 2016; Li et aal., 2016, 2015[15al., 2016; Ma et aal., 2016; Shi et al., 2016; Sun et al., 2016; Tian etal., 2016; Wang etal., 2016, 2015[29al., 2016, 2019[26al., 2016; Wu et aal., 2016; Yang anal., 2016; Yang etal., 2016; Zhang eal., 2016). Ageal The authors declare no conflict of interest."} +{"text": "In the editorial section, Yot Teerawattananon et al. (526) point out that future procurement of vaccines for COVID-19 needs to account more accurately for the doses that can be delivered. Shaoshan Liu et al. (527) explore the potential of autonomous mobile clinics for provision of services in rural and remote areas.Vijay Shankar Balakrishnan (530\u2013531) reports on investigations into a current outbreak of acute hepatitis in children and how this process may prompt research on other causes. Balakrishnan interviews Y\u014dhei Sasakawa (532\u2013533) about his work in global health philanthropy with a focus on leprosy.Heather R Chamberlain et al. (562\u2013569) propose a method to estimate the feasibility of physical distancing.Hirotsugu Aiga et al. (534\u2013543) examine microbial contaminants of water from improved sources.Raquel Burgess et al. (578\u2013580) draw attention to the need to support research staff.Israel J\u00fanior Borges do Nascimento et al. (544\u2013561) review evidence of an infodemic.Erika Siu and Anne Marie Thow (570\u2013577) show examples of improved earmarking of taxes for health."} +{"text": "In the editorial section, Kidist Kebede Bartolomeos (414) makes the case for violence prevention. Matthew Myers Griffith et al. (415) argue for more investment in applied epidemiology.Lynn Eaton (418\u2013419) reports on collaboration needed to ensure water, sanitation and hygiene in humanitarian emergencies. Andrei Shukshin talks to Gulnaz Azhymambetova (420\u2013421) about empowering nurses to deliver better patient care in Kyrgyzstan.Mattias Schedwin et al. (422\u2013435) analyse survey and conflict data by province. Basilua Andre Muzembo et al. (447\u2013458) review the evidence for test performance.Divya Mecheril Balachandran et al. (436\u2013446) assess indicators for near miss and maternal deaths.Azza Sarfraz et al. (462\u2013464) call for a health-sector response to gender-based violence.Bolajoko O Olusanya et al. (459\u2013461) describe an inclusive development and education agenda."} +{"text": "Achtergrond: Momenteel is er nauwelijks sprake van arbocuratieve samenwerking tussen de eerstelijns- en bedrijfsgezondheidszorg. Waar eerdere initiatieven tot verbetering vooral gericht waren op de huis- en bedrijfsarts, onderzoeken we in deze bijdrage welke rol praktijkondersteuners van de huisarts (POH-ggz en POH-S) en van de bedrijfsarts (POB) voor zichzelf zien bij multiproblematiek. Tevens exploreren we welke belemmeringen er zijn voor arbocuratieve samenwerking door praktijkondersteuners bij multiproblematiek.Methode: We hebben drie focusgroepgesprekken uitgevoerd met zeven POH\u2019s-ggz, elf POH\u2019s\u2011S en acht POB\u2019s\u00a0\u2013 26\u00a0praktijkondersteuners in totaal.Resultaten: De praktijkondersteuners in ons onderzoek komen tijdens hun werk in aanraking met multiproblematiek. POH\u2019s en POB\u2019s zien een rol voor zichzelf weggelegd bij het bespreken en het bieden van ondersteuning bij respectievelijk werk- en priv\u00e9gerelateerde klachten. Daarbij erkennen ze, waar nodig, het belang van arbocuratieve samenwerking om goede zorg te leveren. Op dit moment vindt er echter geen directe samenwerking plaats op het niveau van de praktijkondersteuner. Belemmeringen hiervoor blijken de formele regels rond taakdelegatie en rolopvatting van de POB, onbekendheid en vooroordelen bij vooral POH\u2019s wat betreft de bedrijfsgezondheidszorg, en praktische barri\u00e8res als de AVG-wetgeving en bereikbaarheid.Conclusie: POH\u2019s en POB\u2019s staan open voor arbocuratieve samenwerking, mits een oplossing gevonden wordt voor deze fundamentele en praktische belemmeringen. Al decennia wordt geprobeerd de samenwerking tussen de bedrijfsgezondheidszorg en curatieve zorg (\u2018arbocuratieve samenwerking\u2019) te bevorderen. De eerste initiatieven waren gericht op het stimuleren van samenwerking tussen de huis- en bedrijfsarts via communicatieformulieren en -protocollen . Later iDoor de scheiding tussen behandeling en werkgerelateerde zorg in Nederland is er in de eerstelijnsgezondheidszorg van oudsher beperkt aandacht voor werk , 9, 10. De laatste jaren zijn diverse nieuwe functies ontstaan waarin, afhankelijk van de opleidingsachtergrond, gedelegeerde taken van de huis- of bedrijfsarts overgenomen worden , 16. BinHet doel van het huidige onderzoek is te exploreren welke rol zowel POH\u2019s als POB\u2019s voor zichzelf zien in de behandeling van multiproblematiek en in het bijzonder van respectievelijk werk- en priv\u00e9gerelateerde klachten. Tevens willen we achterhalen wat hun ervaringen zijn met arbocuratieve samenwerking. Hiermee hopen we nieuwe wegen te vinden voor samenwerking tussen de eerstelijns- en de bedrijfsgezondheidszorg.In dit onderzoek is gebruikgemaakt van de focusgroepmethode . Op dezeDit onderzoek is goedgekeurd door de Medisch-Ethische Toetsingscommissie van het VUmc, onder nummer\u00a02021.0332.In 2021 zijn drie focusgroepen gehouden met 26\u00a0praktijkondersteuners, respectievelijk met zeven POH\u2019s-ggz, elf POH\u2019s\u2011S en acht POB\u2019s. Deelnemers zijn geworven via de klankbordgroep van het overkoepelende onderzoek waartoe dit deelonderzoek behoort . AfgevaaAlle focusgroepen hadden dezelfde opzet: na een introductie en kennismakingsronde volgde een korte uitleg over de achtergrond van het onderzoek. Vervolgens vond het groepsinterview plaats, gestructureerd aan de hand van een casus van een fictieve, mannelijke automonteur van 52\u00a0jaar met multiproblematiek, die een hartinfarct heeft gehad en na een geleidelijke re-integratie sinds vier maanden weer volledig aan het werk is. Deze werkende pati\u00ebnt komt op controle bij de POH\u2011S, is doorverwezen naar de POH-ggz of verschijnt op het spreekuur van de POB vanwege zorgen over een hoge bloeddruk, problemen in de priv\u00e9situatie en hoge werkdruk. Zie kader\u00a01 voor de casusbeschrijving. De casus werd besproken aan de hand van drie thema\u2019s waaraan vragen gekoppeld waren over: (1)\u00a0herkenning van multiproblematiek tijdens consulten, (2)\u00a0de rol van de praktijkondersteuner bij multiproblematiek en (3)\u00a0ervaringen met arbocuratieve samenwerking. Alle focusgroepen werden begeleid door auteurs EV en FS. De bijeenkomsten duurden anderhalf uur. Vanwege de COVID-19-pandemie vonden de bijeenkomsten online plaats, via MS Teams of Zoom.De focusgroepgesprekken zijn met toestemming van de deelnemers opgenomen en getranscribeerd, waarbij alle deelnemers geanonimiseerd zijn. De transcripten zijn in een eerste stap van open coderen door autPeter is onderhoudsmonteur (52\u00a0jaar) en heeft een jaar geleden een hartinfarct gehad. Hij heeft al jaren een hoge bloeddruk en denkt dat werkdruk mede veroorzaker is van het hartinfarct. Na een geleidelijke re-integratie is hij sinds vier maanden weer volledig aan het werk.(POH-S: Voor een controle-afspraak komt Peter bij je in de praktijk.) (POB: Peter komt bij jou op het preventief spreekuur voor een consult.) Peter geeft tijdens het consult aan dat hij zich zorgen maakt over hoe lang hij dit werk nog vol kan houden. Hij heeft het gevoel dat hij weer in zijn oude werkpatroon terugvalt en dat daarmee ook de werkdruk toeneemt. Hierdoor komt hij vaak oververmoeid thuis. Thuis heeft hij ook nauwelijks rustmomenten. Daarnaast maakt hij zich veel zorgen over zijn gezondheid. Zijn bloeddruk is nu onder controle, maar door de hoge werkdruk en zijn thuissituatie is Peter bang dat hij weer last krijgt van een hoge bloeddruk.Hieronder bespreken we de resultaten van de focusgroepen aan de hand van de drie thema\u2019s (1)\u00a0herkenning van multiproblematiek tijdens consulten, (2)\u00a0de rol van de praktijkondersteuner bij multiproblematiek en (3)\u00a0ervaringen met arbocuratieve samenwerking.\u2018Het overzicht hebben ze [\u2026] vaak niet, ze weten eigenlijk ook niet precies waardoor ze ziek zijn geworden. Ze kunnen heel veel dingen opnoemen, maar wat nou de directe aanleiding is geweest is heel ingewikkeld en die is ook niet te vinden. Dus de oplossing is daardoor ook niet zo heel makkelijk h\u00e8? Het cre\u00ebren van overzicht en het geven van uitleg, dat [\u2026] vind ik vaak de belangrijkste dingen in zo\u2019n\u00a0gesprek. Dat je kijkt van, wat is er nou allemaal aan de hand?\u2019 (POH-ggz)\u2018Dan is het een optelsom van problemen op het werk, onhandig gedrag, [\u2026] stresseten en ontregeling en daarmee is de cirkel rond. Dus dan kan ik wel gaan trekken aan die diabetes, maar er is meer aan de hand. En het \u00e9\u00e9n voedt het ander en het is dan zo\u2019n\u00a0vicieuze cirkel die de verkeerde kant op wil.\u2019 (POH-S)Zowel POH\u2019s als POB\u2019s beseffen dat er bij de pati\u00ebnten of werknemers die op hun consult komen soms problemen spelen op meerdere levensgebieden. Dit komt volgens hen vaker voor als er sprake is van een lagere sociaaleconomische status (SES). Het gaat dan om een combinatie van problemen, zoals gezondheidsklachten, werk en financi\u00ebn, maar ook sociale of relatieproblematiek en problemen rond kinderen of mantelzorg. Volgens de POH\u2019s en POB\u2019s is multiproblematiek complex omdat het ene probleem het andere probleem in de hand werkt of versterkt. Een POB noemt het \u2018lastig dat je niet altijd invloed hebt op alle factoren waar men tegenaan loopt\u2019. Een POH-ggz en een POH\u2011S geven aan dat de cli\u00ebnt bij multiproblematiek vaak het overzicht mist en dat het daarom belangrijk is om alle problemen in kaart te brengen om hulp te kunnen bieden:Tabel\u00a0Als volgende stap in het aanpakken van multiproblematiek zien de POH\u2019s en POB\u2019s het als hun taak om te kijken waar zijzelf en waar anderen ondersteuning kunnen bieden, zodat de pati\u00ebnt of werknemer voldoende ondersteund wordt om weer in balans te komen. Hierin ervaren de POH\u2019s\u2011S en POB\u2019s wel beperkingen in hun rol: enerzijds door een tekort aan consulttijd, anderzijds door hun beperkte invloed op problemen op andere levensgebieden zie tab.\u00a0. Deze beZoals ge\u00efllustreerd in tab.\u00a0Waarom is er op dit moment geen sprake van samenwerking? Tabel\u00a0Vooral voor de POH\u2019s zijn vooroordelen over en onbekendheid met de bedrijfsgezondheidszorg barri\u00e8res voor het aangaan van arbocuratieve samenwerking. In de focusgroepen gaven ze aan niet bekend te zijn met de collega\u2019s van de bedrijfsgezondheidszorg en ook niet goed te weten wat hun taken en verantwoordelijkheden zijn. \u2018Onbekend maakt onbemind\u2019 werd vaker genoemd als reden voor het gebrek aan samenwerking door de POH\u2019s, wat ook herkend werd door de POB\u2019s. Het is voor de POH\u2019s niet duidelijk wat er bijvoorbeeld met de opgevraagde informatie wordt gedaan. Sommigen maken zich zorgen, vooral over de (on)partijdigheid van de bedrijfsarts. De POH\u2019s-ggz gaven daarbij aan dat ze ervaren dat bedrijfsartsen hen niet altijd serieus nemen en dat deze ook niet weten welke rol de POH-ggz precies speelt.Ten slotte vertelden zowel POH\u2019s als POB\u2019s dat arbocuratieve samenwerking bemoeilijkt wordt door praktische barri\u00e8res als onderlinge onbereikbaarheid en de AVG-wetgeving, waardoor eerst toestemming van de pati\u00ebnt of werknemer gevraagd moet worden voor het uitwisselen van informatie. Deze barri\u00e8res verhogen de drempel om de samenwerking op te zoeken.kunnen aangaan , willen aangaan en \u2013\u00a0meer fundamenteel\u00a0\u2013 of ze de samenwerking wel mogen (taakdelegatie tussen bedrijfsarts en POB) en moeten aangaan (hoort deze bij de rol van de POB?). Waar de eerste twee afwegingen ook werden gezien in eerder onderzoek onder huis- en bedrijfsartsen [De POH\u2019s en POB\u2019s in ons onderzoek werken op dit moment niet samen met collega\u2019s uit respectievelijk de bedrijfs- of eerstelijnsgezondheidszorg als er sprake is van multiproblematiek. Hoewel zowel POH\u2019s als POB\u2019s openstaan voor meer samenwerking, zijn belemmeringen terug te voeren op de vraag of ze deze samenwerking wel fsartsen , 7, rakeVoor zover wij weten is de rol van praktijkondersteuners op dit gebied niet eerder onderzocht. Via focusgroepen is ge\u00ebxploreerd welke rol POH\u2019s\u2011S, POH\u2019s-ggz en POB\u2019s voor zichzelf zien bij het behandelen van multiproblematiek en bij arbocuratieve samenwerking. Hiermee werden deze thema\u2019s vanuit meerdere perspectieven belicht. Een eventuele beperking is dat de deelnemende POH\u2019s\u2011S en POB\u2019s aan het eind van hun opleiding zaten, waardoor hun antwoorden mogelijk niet het handelen en denken van de gehele (meer ervaren) beroepsgroep weerspiegelen. Hoewel dit bij de POB\u2019s niet naar voren kwam, vonden enkele POH\u2019s\u2011S het door een gebrek aan ervaring lastig om sommige vragen \u2013\u00a0vooral die over het omgaan met multiproblematiek\u00a0\u2013 te beantwoorden. Ze houden zich tijdens de consulten nog vooral bezig met het eigen taakgebied en hebben daardoor minder oog en tijd voor andere thema\u2019s. Mogelijk voelen meer ervaren POH\u2019s\u2011S zich meer toegerust om binnen de consulttijd ook aandacht te besteden aan problemen op het gebied van werk. Een tweede mogelijke beperking is dat de drie leden van de klankbordgroep die de focusgroep organiseerden en deelnemers wierven een sterke mening hebben over arbocuratieve samenwerking en daarmee minder representatief zijn. Hoewel dit zal gelden voor de drie klankbordleden, zal hier in mindere mate sprake van zijn bij de deelnemers, die vanuit hun opleiding (POH\u2011S en POB) of vanuit hun betrokkenheid bij de intervisiegroep (POH-ggz) (verplicht) deelnamen.In ons onderzoek bleken de Werkwijzer Taakdelegatie en het ontbreken van een BIG-registratie belemmeringen voor arbocuratieve samenwerking. Een mogelijke oplossing om tot meer arbocuratieve samenwerking tussen POH\u2019s en POB\u2019s te komen, is door het gesprek uit de medische hoek te halen en in plaats daarvan een mogelijkheid te bieden om laagdrempelig te communiceren met een collega uit het andere zorgdomein. Hoewel de cocreatie van oplossingen niet het doel was van het onderhavige onderzoek, werd tijdens de focusgroepbijeenkomsten gesuggereerd dat overleg tussen praktijkondersteuners niet hoeft te gaan over medische feiten, maar kan dienen \u2018om gewoon je zorgen uit te spreken over wat je gehoord hebt en waarvan je vindt dat er iets mee moet gebeuren\u2019. Volgens deze opvatting is arbocuratieve samenwerking niet beperkt tot het delen van medische informatie, maar is het ook een manier om te kijken wat de werkende pati\u00ebnt nodig heeft om op meerdere levensgebieden gezond te functioneren. Dit sluit goed aan bij ontwikkelingen waarbij gezondheid breder wordt gezien dan een medisch te duiden probleem, zoals bij de theorie over Positieve Gezondheid . BovendiOnbekendheid met elkaars werk en zeker vooroordelen zijn hardnekkige belemmeringen voor arbocuratieve samenwerking, wat ook in onderzoek onder huis- en bedrijfsartsen naar voren kwam , 7. ReceEen andere belemmering voor arbocuratieve samenwerking tussen de eerstelijns- en de bedrijfsgezondheidszorg heeft betrekking op het ontbreken van structurele financiering om deze samenwerking aan te gaan , 25, 26.Ten slotte is nader onderzoek nodig naar de manier waarop huisartsen en bedrijfsartsen aankijken tegen arbocuratieve samenwerking tussen POH\u2019s en POB\u2019s. Naast de vraag of en welke zorgprofessionals willen samenwerken, is het ook relevant om te onderzoeken of werkende pati\u00ebnten behoefte hebben aan deze samenwerking. Tot zover is er weinig onderzoek gedaan vanuit het pati\u00ebntperspectief, vooral bij werkenden met een lagere SES \u201329. EerdDit onderzoek laat zien dat praktijkondersteuners binnen de eerstelijns- en bedrijfsgezondheidszorg niet of nauwelijks samenwerken, ook niet als er sprake is van multiproblematiek. Hoewel zij hier wel voor openstaan, zijn er naast belemmeringen als de AVG-wetgeving en bereikbaarheid, ook meer fundamentele belemmeringen , die moeten worden geslecht wil men POH\u2019s en POB\u2019s stimuleren tot meer samenwerking en afstemming in het belang van de werkende pati\u00ebnt. Mogelijke oplossingen zijn het uit de medische hoek halen van samenwerking op het niveau van de POH en POB, het besteden van aandacht aan arbocuratieve samenwerking en de bedrijfs- of eerstelijnsgeneeskunde in de opleiding tot POH-S/ggz en POB, het aanpassen van consultprotocollen of het organiseren van gezamenlijke bijeenkomsten voor POH\u2019s en POB\u2019s. Voor het oplossen van structurele belemmeringen, zoals de AVG-wetgeving en financiering, is op politiek niveau overleg nodig."} +{"text": "De toename van de levensverwachting bij de geboorte is het resultaat van de op- en neergang van sterfte aan een groot aantal afzonderlijke ziekten. Dat zoveel ziekten een patroon van opkomst en neergang vertonen, berust op het feit dat zowel opkomst als neergang veelal een direct of indirect gevolg zijn van sociaaleconomische ontwikkelingen. Deze leiden enerzijds tot blootstelling aan nieuwe gezondheidsrisico\u2019s, anderzijds tot meer mogelijkheden om gezondheidsrisico\u2019s te bestrijden, in de vorm van publieke gezondheidszorg of medische zorg. Dit paradoxale verschijnsel wordt ge\u00efllustreerd aan de hand van historische Europese voorbeelden, waarbij vervolgens de vraag aan de orde komt hoe de Nederlandse ervaringen op dit vlak zich verhouden tot die van andere landen, in het bijzonder Zweden, dat al gedurende lange tijd een van de meest succesvolle landen is op het gebied van preventief gezondheidsbeleid. Alleen rond het midden van de twintigste eeuw streefde Nederland Zweden voorbij, in het bijzonder wat betreft het verlagen van de zuigelingensterfte, maar sindsdien is Nederland weer teruggezakt in een Europese \u2018subtop\u2019, onder meer door een weinig doortastend antirookbeleid. Dit wijst erop dat de publieke gezondheidszorg in Nederland beter moet kunnen door op zoek te gaan naar een succesformule die past bij de gezondheidsproblemen van de eenentwintigste eeuw. Weinig andere historische ontwikkelingen hebben ons leven zo sterk veranderd als de stijging van de levensverwachting. In Nederland kan de ontwikkeling van de levensverwachting bij de geboorte worden gevolgd vanaf de tweede helft van de negentiende eeuw, en sindsdien is de gemiddelde levensverwachting bij de geboorte gestegen van ongeveer 40\u00a0tot ruim 80\u00a0jaar.In fig.\u00a0De toename van de levensverwachting heeft een complexe achtergrond, want hij is het resultaat van de op- en neergang van sterfte aan een groot aantal afzonderlijke gezondheidsproblemen. Zoals in het hierna volgende zal blijken, werden in een soort \u2018Echternach-processie\u2019 steeds twee stappen vooruit gedaan, wanneer de sterfte aan enkele ziekten afnam, maar werd vaak ook weer een stap terug gedaan, doordat intussen een nieuwe ziekte opkwam. In deze bijdrage zal ik dit fenomeen van opkomst en neergang van ziekten bespreken, waarbij ik ook zal ingaan op de vraag hoe het komt dat Nederland internationaal soms wel, soms niet voorop liep in het bevorderen van de volksgezondheid.Van 43\u00a0ziekten of andere gezondheidsproblemen kan de langetermijntrend in Europa worden nagegaan, en bij niet minder dan 34\u00a0van deze ziekten is een patroon van opkomst en neergang te zien geweest. Terwijl sommige \u2018opkomsten en neergangen\u2019 zich over vele millennia uitstrekten, duurde het voor andere slechts twee decennia om zich te ontvouwen. Sommige ziekten bereikten hun hoogtepunt in de zeventiende of achttiende eeuw, andere deden dat in de negentiende of twintigste eeuw. De sterfte aan weer andere ziekten stijgt nog steeds en zal hopelijk in de toekomst gaan dalen .Het feit dat zoveel ziekten een patroon van opkomst en neergang hebben vertoond, wijst op een gemeenschappelijke verklaring, en die is te vinden in de samenhang met sociaaleconomische ontwikkelingen in de brede zin van het woord. Als gevolg van een onvermoeibare drang naar verbetering van de levensstandaard wordt de mensheid voortdurend geconfronteerd met nieuwe ziekterisico\u2019s wanneer nieuwe gebieden worden ontsloten, nieuwe productiewijzen in gebruik worden genomen of nieuwe leefgewoonten ontstaan , 4. OmdaDe opkomst van infectieziekten, zoals dysenterie en andere darminfecties, begon waarschijnlijk met de komst van de landbouw in het neolithicum (rond 6.500 voor Christus in Europa), als gevolg van het feit dat mensen dichter op elkaar gingen wonen. Nadat door de verstedelijking de blootstelling aan darminfecties eerst nog veel verder was toegenomen, werden deze ziekten pas in de negentiende eeuw teruggedrongen door maatregelen zoals het aanleggen van drinkwaterleidingen en riolering, die zonder technologische vooruitgang en een toegenomen welvaartsniveau niet mogelijk waren geweest [cordons sanitaires [Een andere ziekte die opkwam met de landbouw is malaria, die ooit endemisch was in Europa, ook in Nederland. Pas vanaf de achttiende eeuw werd malaria teruggedrongen, als gevolg van drainage van moerassen, betere voeding, gebruik van kinine, en veel later ook van gerichte bestrijdingscampagnes, onder meer met DDT . Ook eennitaires .Door de industrialisatie en urbanisatie van Europa kwamen vervolgens weer nieuwe ziekten op, waaronder tuberculose. De geschiedenis van deze ziekte is bekend geworden door het werk van Thomas McKeown, die liet zien dat de daling van tuberculose in Engeland al lang voor de introductie van antibiotica was begonnen. Hij concludeerde daaruit dat medische zorg en ook publieke gezondheidszorg minder belangrijk zijn geweest voor de stijging van de levensverwachting, dan veel mensen in zijn tijd dachten . InmiddeNiet alleen tuberculose, ook veel andere ziekten die met industrialisatie en urbanisatie samenhingen, raakten uiteindelijk op hun retour. Dat gold voor darminfecties als cholera en buiktyfus, voor kinderziekten als kinkhoest en difterie, voor voedingsdefici\u00ebnties als pellagra en rachitis, voor de ziekten die tot moeder- en zuigelingensterfte leiden, enzovoort . In veelTer illustratie is in fig.\u00a0De ziekten van industrialisatie en urbanisatie werden vervolgens ingeruild voor weer nieuwe ziekten, die opnieuw vaak samenhingen met sociaaleconomische ontwikkelingen, in de vorm van een sterk toegenomen welvaartsniveau en de daarmee verband houdende productie- en consumptiepatronen. Na de Tweede Wereldoorlog ontstonden in heel Europa grote epidemie\u00ebn van ischemische hartziekte, diabetes, diverse vormen van kanker en verkeersongevallen. Net als malaria en tuberculose zijn ook dit in zekere zin \u2018ziekten van de vooruitgang\u2019.Kanker is hiervan een goed voorbeeld. Veel vormen van kanker zijn in de loop van de twintigste eeuw sterk toegenomen door veranderingen in leefomstandigheden en gedrag, als gevolg van wat we meestal als vooruitgang zien. Denk aan de grootschalige, industri\u00eble productie van veel wat goed voor ons is, maar ook van sigaretten, alcoholische dranken en asbest, die kanker veroorzaken. En denk aan uitstel van het krijgen van kinderen, zodat er tijd is voor het volgen van een hogere opleiding, maar ook het risico op borstkanker toeneemt .Zoals uit fig.\u00a0Een ander belangrijk voorbeeld van een aandoening die opkwam met de naoorlogse welvaart, maar waarvan de sterfte inmiddels sterk is gedaald, is ischemische hartziekte. Net als bij kanker is de daling deels toe te schrijven aan bestrijding van risicofactoren, in dit geval roken, hoge bloeddruk en een hoog cholesterolgehalte, en deels aan verbeterde behandeling, waardoor de overleving na een hartinfarct enorm is verbeterd . DiabeteHoewel sociaaleconomische ontwikkeling niet alleen aan de opkomst, maar indirect ook aan de neergang van veel aandoeningen ten grondslag ligt, speelt effectief menselijk ingrijpen bij dat laatste in veel gevallen een directe en doorslaggevende rol. In tab.\u00a0Wat de ontwikkeling van de levensverwachting betreft deden de meeste continentaal-Europese landen het gedurende de negentiende en twintigste eeuw aanzienlijk minder goed dan de Scandinavische landen, in het bijzonder Zweden. Zoals hierboven al werd aangegeven gold dat ook voor Nederland, althans in de tweede helft van de negentiende eeuw en in de laatste decennia van de twintigste eeuw, maar niet rond het midden van de twintigste eeuw, toen Nederland zelfs in enkele kalenderjaren de Europese recordhouder was. Wat verklaart de opmerkelijke prestatie van Nederland in de eerste helft van de twintigste eeuw, en de relatieve achteruitgang in de tweede helft?In de negentiende eeuw was het met de volksgezondheid in Nederland niet goed gesteld. West-Nederland kende toen een zeer hoge sterfte, wat overigens al in de achttiende eeuw bekend was. Volgens de Britse demograaf Thomas Malthus (1766\u20131834) was Nederland zelfs \u2018het graf van Duitsland\u2019, omdat zoveel Duitse immigranten er hun vroegtijdige dood vonden. Deze erbarmelijke situatie stond in schril contrast met het roemrijke verleden van Nederland: in de zeventiende eeuw was de Republiek der Zeven Verenigde Nederlanden het meest welvarende, verlichte en machtige land van Europa. Religieus ge\u00efnspireerde scholingscampagnes tijdens en na de Tachtigjarige Oorlog 1568\u20131648) hadden ook het geletterdheidsniveau naar recordhoogten gebracht. In het begin van de achttiende eeuw verloor de Nederlandse Republiek haar dominante positie in de wereldhandel echter aan Groot-Brittanni\u00eb, en de daaruit voortvloeiende achteruitgang van de economie werd nog verergerd door de schade die werd aangericht door de Napoleontische oorlogen in het begin van de negentiende eeuw [48 haddenMeer specifiek was de hoge sterfte in Nederland in de negentiende eeuw waarschijnlijk te wijten aan een combinatie van hoge bevolkingsdichtheid en gebrek aan goed drinkwater, die beide het risico op infectieziekten vergrootten. Verder kwamen maatregelen op het gebied van de publieke gezondheid in Nederland traag op gang. Deze situatie veranderde pas rond 1870, toen als eerste teken van vooruitgang op het terrein van de volksgezondheid de kindersterfte begon af te nemen, waarschijnlijk meer als gevolg van culturele veranderingen , dan als gevolg van economische groei. Een hoog niveau van geletterdheid \u2013\u00a0een overblijfsel van het glorieuze verleden van Nederland\u00a0\u2013 zal deze snelle verbeteringen mede mogelijk hebben gemaakt .In de eerste helft van de twintigste eeuw werden trends in de levensverwachting nog sterk bepaald door trends in zuigelingensterfte. Door een opmerkelijke daling van de zuigelingensterfte tot internationaal gezien zeer lage niveaus zie fig.\u00a0 steeg deDat Nederland \u2013\u00a0in het derde kwart van de negentiende eeuw nog achterlopend op meer geavanceerde Europese landen\u00a0\u2013 erin slaagde om zich bij de Scandinavische landen aan de top van de Europese ranglijst te voegen, zal deels verklaard worden door de snelle groei van de Nederlandse economie, zowel voor, tijdens als direct na de Eerste Wereldoorlog. Dit kwam door een combinatie van factoren, waaronder het feit dat Nederland profiteerde van snelle economische groei in het directe achterland, namelijk de belangrijkste handelspartner Duitsland . Een sneHelaas verloor Nederland zijn voorsprong in de jaren tachtig en negentig van de twintigste eeuw, toen in sommige leeftijdsgroepen in Nederland een volledige stagnatie van de sterftedaling optrad, terwijl veel andere Europese landen hun snelle sterftedaling voortzetten. Deze stagnatie was zowel bij de allerjongsten , als bij de alleroudsten (sterfte onder 80-plussers) te zien.De verloskundige beroepsgroepen schreven aanvankelijk de hoge perinatale sterfte toe aan een meer volledige registratie in Nederland, maar dit standpunt werd onhoudbaar met de publicatie van internationaal geharmoniseerde cijfers, die bevestigden dat Nederland ongunstig afstak bij andere Europese landen . Meer geDe stagnatie van de sterftedaling onder ouderen was deels het gevolg van een gebrek aan vooruitgang bij het terugdringen van roken, dat extreem hoge niveaus had bereikt onder de Nederlandse geboortecohorten die in de jaren tachtig en negentig overleden (zie hierboven). Een vergelijkbare stagnatie van de sterftedaling onder ouderen deed zich voor in Denemarken, ook deels als gevolg van een historisch hoge prevalentie van roken , 25. EenBehalve een tekortschietend antirookbeleid hebben vermoedelijk ook budgettaire beperkingen in de gezondheidszorg een rol gespeeld bij de stagnatie van de sterftedaling onder ouderen in Nederland. Deze budgettaire beperkingen leidden in de jaren tachtig en negentig tot een minder snelle groei van de zorguitgaven in Nederland dan in andere Europese landen, maar remden ook de introductie van nieuwe effectieve behandelingen af. Na 2002, toen de regering had besloten de budgettaire beperkingen op te heffen, begon zowel het gebruik (en de kosten) van de gezondheidszorg als de levensverwachting weer snel te stijgen .Voor de publieke gezondheidszorg in Nederland heeft deze geschiedenis zowel bevredigende als teleurstellende kanten. Het is, om met het goede nieuws te beginnen, goed om nog eens vast te stellen dat de publieke gezondheidszorg een cruciale rol heeft gespeeld bij de toename van de levensverwachting bij de geboorte, niet alleen in het verre verleden, maar ook in de laatste decennia van de twintigste eeuw. Het ging hierbij overigens om publieke gezondheidszorg in de ruime zin van het woord, dus niet beperkt tot instellingen met publieke gezondheidszorg in hun missie, en al helemaal niet tot professionals met een opleiding op dit terrein. De aanleg van drinkwaterleidingen en riolering, maatregelen op het gebied van de verkeersveiligheid, en het heffen van accijnzen op alcohol en tabak vallen immers geheel of grotendeels binnen het domein van andere instellingen of disciplines.Ook het feit dat de publieke gezondheidszorg de verantwoordelijkheid voor successen in de volksgezondheid tegenwoordig moet delen met de medische zorg moet tot enige bescheidenheid manen. In tegenstelling tot wat velen op gezag van McKeown nog steeds aannemen, is medische zorg met al zijn technisch vernuft, in combinatie met een verzekeringsstelsel dat een goede financi\u00eble toegankelijkheid regelt, uitgegroeid tot een effectief instrument om de gezondheid van de hele bevolking te bevorderen. De spectaculaire daling van de sterfte aan ischemische hartziekte sinds de jaren zeventig van de twintigste eeuw was zonder de cardiologie niet mogelijk geweest. Voork\u00f3men is nog steeds beter dan genezen, en vaak ook goedkoper, maar helaas is preventie lang niet altijd mogelijk, en is er ook niet altijd voldoende draagvlak voor preventief beleid, en dan is het goed dat er ook curatieve mogelijkheden zijn.Voor de publieke gezondheidszorg in Nederland waren, achteraf gezien, de decennia rond het midden van de twintigste eeuw de \u2018gouden jaren\u2019, althans wanneer we het succes van deze sector afmeten aan uitkomstmaten als zuigelingensterfte en levensverwachting. Op een of andere manier is het echter niet gelukt dit succes goed vast te houden, dat wil zeggen een succesformule te vinden die past bij de gezondheidsproblemen van de huidige tijd. Dat blijkt ook uit een recent onderzoek waarin werd nagegaan hoe de resultaten van het Nederlandse gezondheidsbeleid zich in de periode 1970\u20132010 verhielden tot die van andere Europese landen. Hierbij werden de nationale prestaties vergeleken op tien verschillende terreinen van preventief gezondheidsbeleid, namelijk roken, alcohol, voeding, zwangerschap en bevalling, jeugdgezondheidszorg, infectieziekten, hypertensie, kankerscreening, verkeersveiligheid en luchtverontreiniging. In de Europese rangorde op basis van een samenvattende score stond Nederland op de vijfde plaats, na Zweden, Noorwegen, IJsland en Finland . De concDe kans lijkt mij groot dat internationaal-vergelijkende onderzoeken van de uitkomsten van het beleid gevoerd tijdens de COVID-19-pandemie tot vergelijkbare conclusies zullen leiden. Over de jaren 2020 en 2021 lag de aan COVID-19 toe te schrijven \u2018oversterfte\u2019 in Nederland op het (West\u2011)Europese gemiddelde, en beduidend boven die in de Scandinavische landen . De publ"} +{"text": "In the editorial section, Nathalie Roebbel et al. (750) outline global research priorities for urban health. Hans Henri P Kluge et al. (751) introduce digital transformation programmes in the WHO Region of Europe. Gary Humphreys (754\u2013755) reports on an initiative to replace tobacco cultivation with healthier crops in Kenya. In the interview section, Maria Munir Yusuf talks to Humphreys (756\u2013757) about her work in Ethiopia to provide shelter, care and livelihoods for women who survive gender-based violence.Phoebe CM Williams et al. (798\u2013807) propose an antibiotic development list for priority pathogens.Michael M Lindeborg et al. (789\u2013797) argue for standard definitions and better treatment options.B\u00e9ninwend\u00e9 Leticia Delphine Sakana et al. (769\u2013776) evaluate use of a routine immunization visit.Mengyuan Pan et al. (758\u2013768) assess national procurement data.Courtney M Yuen et al. (777\u2013788) review the evidence for integrated care models.Steph H Tan et al. (808\u2013814) make the case for saliva-based tests. John J Reilly et al. (815\u2013824) explain how quantifying inactivity can improve health policies."} +{"text": "Het doel van dit vragenlijstonderzoek was om in kaart te brengen op welke manieren en op welke momenten mensen met diabetes mellitus type 2 willen participeren bij de ontwikkeling en toepassing van e-health, en welke factoren daarop van invloed zijn.Via verschillende online platforms en de nieuwsbrief van de Diabetesvereniging Nederland is een digitale vragenlijst verspreid met zowel gesloten als open vragen. Informatie werd verzameld over: 1) bereidheid tot participatie; 2) voorkeuren over de vorm van participatie; 3) be\u00efnvloedbare factoren voor participatie, zoals motivatie, competentie, middelen, sociale invloed en uitkomstverwachtingen; 4) achtergrondkenmerken.Er zijn 160 vragenlijsten geanalyseerd. Ruim 75% van de respondenten heeft interesse in pati\u00ebntparticipatie. De meeste respondenten prefereren solistische participatiemethoden boven groepsparticipatie, respectievelijk 93% en 46%. De helft denkt voldoende kennis te hebben om mee te kunnen doen aan pati\u00ebntparticipatie en 40% denkt een waardevolle inbreng te kunnen hebben. Als vergoeding wensen deelnemers vooral het gratis gebruik van nieuwe technologie. Omdat mensen verschillen in hun voorkeuren voor momenten en manieren van participatie, is het aan te bevelen daarvoor verschillende vormen van participatie en vergoedingen aan te bieden tijdens het gehele proces van ontwikkeling tot toepassing van e-health.1 Het aantal mensen met deze vaak leefstijlgerelateerde aandoening neemt snel toe door vergrijzing en ongezonde leefgewoonten.2-4 Dit zorgelijke vooruitzicht benadrukt het belang van preventie, gezondere leefstijl en zelfmanagement om de prevalentie van DMT2 terug te dringen en om de behandeling van DMT2 te ondersteunen.5 De komende jaren zal de zorg van een chronisch zieke meer naar de thuissituatie gaan verschuiven, waardoor zelfmanagement, eigen regie en autonomie belangrijker worden.3Diabetes mellitus type 2 (DMT2) is een van de meest voorkomende chronische aandoeningen in Nederland, met een enorme impact op kwaliteit van leven en zorgkosten.6 Dit biedt de mogelijkheid om zorg en begeleiding digitaal en op afstand te organiseren.7 E-health heeft grote potentie om mensen met diabetes te ondersteunen bij zelfmanagement in de thuissituatie en om op een andere manier samen te werken met zorgprofessionals. Interventies gericht op aanpassing van leefstijl met ondersteuning van e-health blijken voor deze pati\u00ebntengroep effectief in het verbeteren van de kwaliteit van leven, HbA1c-niveaus, bloeddruk en body mass index (BMI).8-11E-health betreft 'het gebruik van nieuwe informatie- en communicatietechnologie ter ondersteuning of verbetering van gezondheid en de gezondheidszorg', zoals gezondheidsapps, platforms of digitale coaches.12-13 Daarnaast speelt ook het type innovator waarmee iemand zich identificeert een rol bij het accepteren en gebruiken van e-health. Rogers heeft vijf groepen gedefinieerd die zich onderscheiden in acceptatie van technologie.14 Deze groepen zijn de innovatoren, pioniers, voorlopers, achterlopers en achterblijvers, waarbij de eerste groep op zoek is naar het nieuwste en de laatste groep liever gebruikt wat ze al hebben.14 Het type innovator kan een rol spelen bij de bereidheid om te participeren in onderzoek naar e-health.Desondanks kunnen pati\u00ebnten verschillende barri\u00e8res ervaren rond het gebruik van e-health. Zo kan het zijn dat ze zich niet vertrouwd voelen met e-health, deze te complex is, niet goed werkt of dat ze geen toegang hebben, of dat de toepassing en het zorgproces niet op elkaar zijn afgestemd.15,16 Pati\u00ebntparticipatie houdt in dat pati\u00ebnten de mogelijkheid krijgen om tijdens het innovatieproces hun kennis en ervaringen met hun eigen aandoening in het dagelijks leven in te brengen en zo bij te dragen aan verbeteringen in de zorg.17 Ervaringen van mensen met een aandoening kunnen erg waardevol zijn, omdat zij op een andere manier naar de zorg kijken dan zorgprofessionals, wetenschappers en ontwikkelaars. Door deze perspectieven te combineren, kan persoonsgerichte zorg ontstaan. Pati\u00ebntparticipatie heeft de afgelopen decennia een enorme ontwikkeling doorgemaakt, waardoor er steeds meer aandacht is voor inbedding van pati\u00ebntperspectieven in onderzoek.18 Pati\u00ebntparticipatie kan de rechtvaardigheid van besluiten verhogen, succesvolle implementatie in de zorg faciliteren en de effectiviteit en doelmatigheid van het zorgsysteem vergroten.17,18 Daarbij verhoogt pati\u00ebntparticipatie de kennis van deelnemers over gezondheid en leefstijl, en activeert ze deelnemers om deze kennis toe te passen bij het bevorderen van gezondheid.19Participatie van pati\u00ebnten bij de ontwikkeling en toepassing van e-health kan zorgen voor meer aansluiting bij hun wensen, behoeften en mogelijkheden, en hiermee de bereidheid tot het gebruik vergroten.16 Deze routekaart bestaat uit vijf fases die samen het innovatieproces vormen, namelijk: 1) contextbepaling , 2) waardespecificatie , 3) ontwerp (ontwerpen van prototype met belanghebbenden), 4) operationalisatie en 5) summatieve evaluatie en continue formatieve evaluatie (verzamelen van kennis om vervolgstappen te kunnen maken). In elk van deze fases kan de pati\u00ebnt een actieve rol spelen.Pati\u00ebntparticipatie kan op verschillende manieren georganiseerd worden in het gehele proces van e-healthontwikkeling tot toepassing in de (zorg)praktijk. De CeHRes-routekaart biedt een stappenplan dat helpt om e-health te cre\u00ebren waarvoor draagvlak is en die aansluit bij de behoeften van gebruikers.20 Deze acht rollen zijn manipulatie, therapie, informatie, consultatie, adviseren, partnerschap, gedeelde macht en regie. Hiervan worden manipulatie en therapie gezien als niet-participatie. De rollen informatie, consultatie en adviseren worden tokenisme genoemd, waarin de burgers een oppervlakkige participatie hebben. Bij de rollen partnerschap, gedeelde macht en regie hebben de participanten steeds meer controle over het onderzoek . Onderzoekers en Ziekenhuisgroep Twente (ZGT) werken samen aan onder andere participatie van mensen met DMT2 bij het innovatieproces van e-health. Er is begonnen met mensen met DMT2 omdat er rond deze aandoening veel e-health ter ondersteuning van leefstijl en zelfmanagement wordt ontwikkeld. Het doel van het project is het ontwikkelen van een persoonsgericht, technologieondersteund aanbod. Dit zou de therapietrouw kunnen verbeteren en kan mensen met DMT2 meer inzicht bieden in de invloed van leefstijl op diabetes. In combinatie met coaching, digitaal of fysiek, geeft dit de mensen met DMT2 mogelijk meer eigen regie, en kan het bovendien de zorgprofessional ondersteunen. Om deze verbeteringen te bewerkstelligen is e-healthontwikkeling nodig die aansluit bij de behoeften en wensen van mensen met DMT2. Participatie van mensen met DMT2 in deze innovatieprocessen kan zorgen voor de meest passende en bruikbare e-healthontwikkelingen. Kennis over de participatiewensen kan belangrijke ondersteuning bieden bij het stimuleren van participatie. Het doel van dit eerste onderzoek binnen dit project is om te beschrijven op welke momenten tijdens het innovatieproces en op welke manieren mensen met DMT2 zouden willen participeren. Om de bereidheid en voorkeuren voor participatie van mensen met DMT2 te onderzoeken heeft een crosssectioneel vragenlijstonderzoek plaatsgevonden. Vanwege de verkennende focus van dit onderzoek is er gekozen voor een vragenlijst om een groep respondenten te bereiken die representatief is voor de populatie mensen met DMT2 in Nederland. Voor de werving van mensen met DMT2 is gebruikgemaakt van verschillende online platforms, zoals LinkedIn, Facebook en Twitter. Daarnaast is er een oproep geplaatst in de online nieuwsbrief van Diabetesvereniging Nederland (DVN) en op het forum van het Diabetes Trefpunt. Inclusiecriteria voor dit onderzoek waren mensen met DMT2 van achttien jaar of ouder die de Nederlandse taal machtig waren. Mensen met ernstige comorbiditeit of psychische stoornissen en/of mensen die geen gebruik kunnen maken van een computer werden ge\u00ebxcludeerd. Het onderzoek werd bij de ethische toetsingscommissie van de Universiteit Twente goedgekeurd (aanvraagnummer 200589). Respondenten kregen toegang tot de online vragenlijst nadat zij toestemming hadden gegeven via een online toestemmingsformulier.De vragenlijst bestond uit 33 vragen verdeeld over vier delen: 16, met antwoordopties in de vorm van een vijfpuntslikertschaal (zeker niet tot en met zeker wel). De fases werden benoemd en uitgelegd aan de respondenten: 1) contextbepaling, informatieverzameling over de eindgebruiker, 2) waardespecificatie, informatieverzameling over wat eindgebruikers belangrijk vinden bij een nieuwe technologie, 3) ontwerp, testen van het prototype en 4) operationalisatie en evaluatie, aangepaste technologie inzetten in de praktijk en evalueren van de technologie. 1. Bereidheid tot participatie op verschillende momenten van een e-healthinnovatieproces: vier vragen gebaseerd op de CeHRes-routekaart20,met antwoordopties in de vorm van een vijfpuntslikertschaal (zeker niet tot en met zeker wel) en drie meerkeuzevragen. Respondenten werd gevraagd of zij individueel of in een groep deel willen nemen aan verscheidene onderzoeksmethoden. De individuele methoden waren een vragenlijst, interviews, het bijhouden van een dagboek of onderdeel zijn van een regelmatig panel. De methoden in een groep betreffen groepsgesprekken met andere mensen met DMT2 of spiegelgesprekken waarbij ook zorgprofessionals aanwezig zijn, deelnemen aan een cli\u00ebntenraad, of aan een werkgroep om met de ontwikkelaar of als medeonderzoeker mee te werken in het hele onderzoek. 2. Voorkeuren met betrekking tot de vorm van participatie: drie vragen gebaseerd op de participatieladder21, met antwoordopties in de vorm van een vijfpuntslikertschaal (zeker niet tot en met zeker wel), drie meerkeuzevragen en twee open vragen naar redenen om wel of niet te participeren in onderzoek.3. Be\u00efnvloedende factoren voor participatie als motivatie, competentie, middelen, sociale invloed en verwachtingen van innovaties: tien vragen gebaseerd op de be\u00efnvloedende factoren voor participatie144. Achtergrondkenmerken van de respondenten: zeven meerkeuzevragen en \u00e9\u00e9n open vraag. Hierin is een vraag meegenomen over het type innovator dat past bij de respondent, met daarin innovatoren , pioniers ('ik probeer graag nieuwe technologie voor diabetes uit'), voorlopers , achterlopers en achterblijvers ('ik gebruik nieuwe technologie niet snel en blijf graag gebruiken wat ik ken').Bij de gesloten vragen konden de respondenten antwoordopties selecteren. In de open vragen behorende bij deel 3 werden de respondenten gevraagd om drie redenen te noemen waarom zij wel of niet zouden willen participeren. Hierbij konden de respondenten vrije tekst invullen. Om de validiteit te vergroten werd de vragenlijst ontwikkeld in goed overleg met een groep van zes onderzoekers, twee medewerkers van het ZGT en twee mensen met DMT2.De vragenlijst werd uitgezet van 4 mei tot en met 10 juli 2020. Het beantwoorden van de vragenlijsten kostte gemiddeld 15 minuten. De gegevensanalyse van de gesloten vragen werd gedaan met SPSS Statistics (versie 24). De analyses betroffen beschrijvende statistiek. Bij iedere vraag werden het aantal respondenten en het percentage dat voor elke antwoordoptie heeft gekozen gerapporteerd. Bij de rapportage van leeftijd en aantal jaren gediagnosticeerd met DMT2 werd een gemiddelde en standaarddeviatie weergegeven. De gegevensanalyse van de open vragen werd gedaan door twee onderzoekers door middel van een inductieve thematische analyse in Excel. Ieder gegeven antwoord werd gecodeerd en aan ieder antwoord werd een thema gekoppeld. Na het coderen werd een vergelijking van de beschrijvende thema's gemaakt. Bij elkaar passende thema's werden gecombineerd, zodat er een codeboom met hoofd- en subthema's ontstond. Sprekende citaten werden uitgekozen om weer te geven in de resultaten. De resultaten hiervan zijn besproken met meerdere onderzoekers om de betrouwbaarheid en validiteit te verhogen.Honderdzestig mensen met DMT2 hebben de vragenlijst volledig ingevuld. De gemiddelde leeftijd was 67 jaar , de jongste respondent was 27 en de oudste 89 jaar. Gemiddeld hebben de respondenten 16,2 jaar DMT2 , vari\u00ebrend van 3 maanden tot 47 jaar. Er hebben 13 respondenten met een migratieachtergrond deelgenomen. Daarnaast is gevraagd welk 'type innovator' bij de respondenten past .kenmerknHet overgrote deel van de respondenten (79% tot 88%) is bereid of heeft de intentie om actief deel te nemen aan het innovatieproces van e-health voor mensen met DMT2. Achtentachtig procent van de respondenten wil 'waarschijnlijk wel' of 'zeker wel' betrokken worden bij het in kaart brengen van het dagelijks leven, de behoeften en de problemen van mensen met DMT2 . Daarnaast zou 79% 'waarschijnlijk wel' of 'zeker wel' willen deelnemen aan het in kaart brengen van de wensen en eisen van mensen met DMT2 met betrekking tot een bepaalde technologie (de waardespecificatie). Aan het testen van prototypen, de ontwerpfase, en aan de operationalisatie en evaluatiefase in de praktijk zou 84% 'waarschijnlijk wel' of 'zeker wel' deelnemen .fases inHonderdvierenveertig respondenten gaven de voorkeur aan solistische participatiemethoden . DaarvanIn de open vragen werd een verdiepende slag gemaakt en benoemden de respondenten redenen om wel of niet te participeren in het innovatieproces en 3. ReEnerzijds gaven respondenten aan het leuk te vinden, nieuwsgierig te zijn en graag ontwikkelingen op het gebied van technologische innovaties te volgen. Net als in voorgaande vragen werden redenen voor 'eigen belang' en 'belang voor anderen' veel genoemd. 'Om te bekijken of de nieuwe technologie iets voor mij of anderen in mijn omgeving is.' Aan de andere kant gaven respondenten aan technologie ook moeilijk te vinden, kan deze invloed hebben op de privacy en werd niet altijd het nut ervan ingezien: 'Ik heb geen interesse als de technologie geen noemenswaardig voordeel voor mij oplevert.' Hiertegenover was het kunnen uitproberen van innovaties een reden om wel te participeren. Meerdere mensen geven aan dat ze graag zelf de technologie thuis testen en toepassen in hun dagelijks leven. Een aantal respondenten geeft duidelijk aan geen proefkonijn te willen zijn. Vooral participeren in groepsverband wordt benoemd als reden om niet te participeren. E\u00e9n respondent benoemde dit als 'de angst om ondergesneeuwd te worden door verbaal sterke mensen'. Meerdere respondenten geven ook aan dat ze niet deel willen nemen aan interviews als ze 'twijfel hebben over de deskundigheid van de onderzoeker' of als ze een andere negatieve invloed ervaren van de onderzoekers of andere respondenten. Aspecten als tijd, reizen en kosten werden ook veelvuldig genoemd als reden om niet te participeren in het innovatieproces: 'Als het te vaak en te veel tijd in beslag neemt.'Tot op heden is er weinig onderzoek gedaan naar momenten tijdens het innovatieproces van e-health en manieren waarop mensen met DMT2 willen participeren. Uit dit onderzoek blijkt dat de bereidheid onder mensen met DMT2 hoog is om te participeren in iedere fase van het innovatieproces, van contextbepaling tot operationalisatie en evaluatie van e-health. De voorkeur gaat uit naar participatie via vragenlijsten, groepsgesprekken en spiegelgesprekken. De meeste respondenten willen deelnemen omdat ze verwachten dat pati\u00ebntparticipatie de diabeteszorg zal verbeteren of hun leven of dat van anderen met diabetes makkelijker zal maken. Daarnaast wordt inbreng van eigen ervaringen, bijdragen aan verbetering van de technologie en interesse in technologische innovaties genoemd als redenen om te participeren. Het gratis gebruik van technologie die ze kunnen testen wordt gezien als vergoedingsmogelijkheid. Misbruik van data, commerci\u00eble doeleinden en kosten werden genoemd als redenen om niet te participeren in het innovatieproces van e-health.17,18 Er is weinig literatuur over de bereidheid en wensen omtrent de vormen van participatie van mensen voorafgaand aan onderzoek. Een eerder onderzoek naar bereidheid tot participatie laat zien dat mensen willen participeren wanneer het onderzoek gerelateerd is aan een behandeling, maar niet wanneer het risico's met zich meebrengt of wanneer ze weerstand voelen tegen onderzoek.23 Een ander onderzoek heeft deelnemers benaderd die niet gereageerd hadden op een uitnodiging tot participatie. Hieruit kwamen vergelijkbare negatieve gevoelens over risico's en weerstand naar boven, maar ook de verwachte tijdsbelasting werd veel genoemd als reden om niet te participeren.24 Dit onderzoek laat vergelijkbare resultaten zien met betrekking tot redenen om wel of niet te participeren en voegt daaraan toe op welke manier en in welke vorm mensen zouden willen participeren.Het nauw betrekken van pati\u00ebnten bij innovatieprocessen is van belang om de meest passende en gewenste innovaties te realiseren.25,26 De zorgprofessionals van mensen met DMT2 kunnen een belangrijke schakel vormen. Kennis en overtuigingen van zorgprofessionals vormen een van de vijf categorie\u00ebn die invloed hebben op de participatie van pati\u00ebnten.27De bereidheid onder mensen met DMT2 om te participeren in iedere fase van het innovatieproces is hoog. Uit dit onderzoek blijkt dat de bereidheid om te participeren in het innovatieproces voor een deel van de respondenten verhoogd kan worden als zorgprofessionals het belangrijk vinden. Daarnaast blijkt ook dat de zelfeffectiviteit van de respondenten laag is. Met andere woorden, ze achten zichzelf niet of nauwelijks in staat om zelf iets in te brengen. Zelfeffectiviteit kan van invloed zijn op de daadwerkelijke participatie.26 Het was wenselijk een hoger aantal respondenten met een migratieachtergrond te bereiken, aangezien DMT2 bij deze populatie relatief veel voorkomt. Met de wervingsmethoden van dit onderzoek werd deze groep niet aangesproken, iets waar vervolgonderzoek naar zal moeten kijken. Daarnaast zal ook rekening moeten worden gehouden met gezondheidsvaardigheden, waarnaar in dit onderzoek geen uitvraag is gedaan. Dit zou een aanbeveling kunnen zijn voor vervolgonderzoek, vooral om te voorkomen dat bepaalde groepen pati\u00ebnten buitengesloten worden bij onderzoek naar innovaties en dat het gebruik van e-health daardoor voor hen niet past of mogelijk is.Aan dit onderzoek deden 160 respondenten mee, voldoende voor een exploratief onderzoek met een deels kwalitatieve focus. Daarnaast laten de achtergrondkenmerken zien dat het om een zeer diverse populatie gaat. Vanwege de diversiteit van de populatie en daarmee lage aantallen van subpopulaties hebben wij geen verbanden gevonden tussen achtergrondkenmerken en de bereidheid tot deelname of wensen voor pati\u00ebntparticipatie. Het grotere aantal mannen en de gemiddeld hogere leeftijd van de respondenten is representatief voor de Nederlandse populatie mensen met DMT2. Het aantal respondenten met een migratieachtergrond is lager.Uit het innovatieprofiel van de respondenten blijkt dat een groot deel graag nieuwe technologie uitprobeert, wat mogelijk tot selectiebias heeft geleid. In vervolgonderzoek zal naar de representativiteit van deze groep gekeken kunnen worden om eventueel aanvullend onderzoek te doen met bijvoorbeeld mensen met een migratieachtergrond of lage gezondheidsvaardigheden. Daarnaast verwachten we dat een groot aantal van de respondenten al positief is over participeren in onderzoek naar e-health omdat zij al deelnemen aan een online vragenlijstonderzoek. Vanwege de COVID-19-pandemie hebben we gekozen voor online verspreiding van de vragenlijst, wat mogelijk tot bias in het type respondenten heeft geleid. Bij de verspreiding van de vragenlijst zijn verschillende media gebruikt, waardoor we niet weten welke kenmerken mensen met DMT2 hebben die niet hebben deelgenomen. Voor het doel van dit onderzoek was geen gevalideerde vragenlijst beschikbaar en daarom is er gebruikgemaakt van een zelfontworpen vragenlijst. De vragenlijst is opgesteld en beoordeeld op begrip door mensen met lage gezondheidsvaardigheden. Daarna is de vragenlijst getest door medewerkers van het ZGT en mensen met DMT2. De huidige wetenschappelijke onderzoeken streven ernaar om er meer respondenten bij te betrekken. Het zou een goede investering zijn om een vragenlijst over wensen voor participatief onderzoek te ontwikkelen en te valideren, zodat onderzoek kan worden gedaan naar de bereidheid en wensen van mensen om te participeren. Verder zou naast kwantitatief onderzoek ook kwalitatief onderzoek moeten worden gedaan. Op die manier kan diepere informatie verkregen worden over manieren en momenten waarop mensen betrokken willen worden. Daarbij kunnen specifieke vragen opgenomen worden om meer te weten te komen over mensen met bijvoorbeeld lage gezondheidsvaardigheden.21 Hun vorm van participatie werd in de meeste gevallen informatie of consultatie genoemd op de participatieladder van Arnstein; een enkele keer was er sprake van partnerschap of regie.20 Onderzoek naar ervaren meerwaarde zoals benoemd door deelnemers aan pati\u00ebntparticipatie onderscheidt vijf belangrijke thema's, namelijk authenticiteit, samenwerking, bevindingen genereren, disseminatie en betrokken willen blijven bij toekomstige onderzoeken.22 We hebben gevraagd naar de manieren waarop mensen betrokken zouden willen worden. Vervolgonderzoek zou ook specifiek aandacht kunnen besteden aan de wensen met betrekking tot samenwerking met onderzoekers en kunnen kijken naar de ervaringen van de daadwerkelijke pati\u00ebntparticipatie. Bij een evaluatie van de ervaringen met pati\u00ebntparticipatie onder mensen met DMT2 kunnen de vijf thema's met betrekking tot ervaring en meerwaarde van pati\u00ebntparticipatie gebruikt en mogelijk verder uitgewerkt worden.Naar aanleiding van dit onderzoek is een aantal aanbevelingen voor de praktijk en vervolgonderzoek opgesteld. Bij het opzetten van een onderzoeks- en innovatieproject voor e-health is het van belang de doelgroep in een zo vroeg mogelijk stadium bij het onderzoek te betrekken en gezamenlijk in kaart te brengen welke rol de doelgroep in de verschillende fases van het innovatieproces speelt. Bij het opzetten van een pati\u00ebntparticipatieonderzoek moet hier goed over worden nagedacht. Veel respondenten willen bijdragen in de vorm van een vragenlijst, groepsgesprek of spiegelgesprek en er zijn minder respondenten ge\u00efnteresseerd in deelname aan panels of andere intensievere vormen. Dit hoeft geen nadeel te zijn, aangezien er voor deze methoden vaak minder mensen nodig zijn. Bij het opzetten van een onderzoek is het aan te bevelen om tijdens de werving ook te kijken naar vormen van participatie. Tijdens de werving is het mogelijk om de deelnemers te laten kiezen tussen gewenste vormen van participatie en mogelijke vergoedingen. Deze keuze maakt pati\u00ebntparticipatie mogelijk interessanter voor de deelnemers. Uit dit onderzoek komt naar voren dat niet alleen financi\u00eble vergoedingen tot de compensatiemogelijkheden behoren, maar ook dat het gratis testen en behouden van technologie als een mogelijke compensatie gezien wordt. De duur van participatie heeft ook invloed op de bereidheid: mensen willen er liever niet te veel tijd aan besteden. Het is aan te bevelen om in kleine groepen korte sessies te organiseren, waarbij technologie kan worden uitgeprobeerd en getest. Het is ook aan te raden om zorgprofessionals bij deze sessies te betrekken. Er zou meer aandacht besteed kunnen worden aan de rol van zorgprofessionals door hen nauw te betrekken bij het werven van deelnemers voor het innovatieproces. Daarnaast kunnen zorgprofessionals een sleutelrol spelen door het belang van participatie te benadrukken en de mensen met DMT2 te enthousiasmeren om aan diverse methoden deel te nemen. Op deze manier zou een bredere groep participanten bereikt kunnen worden. In samenwerking zouden bepaalde doelgroepen specifiek benaderd kunnen worden, zoals mensen met een migratieachtergrond of lage gezondheidsvaardigheden. In dit onderzoek is niet specifiek gevraagd naar de mate waarin met onderzoekers wordt samengewerkt en naar de complexiteit van de wijze van participeren. De samenwerking kan vari\u00ebren van respondent zijn tot het voeren van de regie. Lemmens et al. laten zien dat mensen die aan onderzoek hebben deelgenomen dat als zeer positief ervaren.Mensen met DMT2 blijken bereid te zijn om op verschillende momenten en manieren samen te werken in innovatieprocessen. Bij pati\u00ebntparticipatie in innovatieprocessen van e-health kunnen ze samen met hun zorgprofessionals een sleutelrol spelen. Mensen met DMT2 willen op verschillende manieren bij het innovatieproces betrokken zijn. Rekening houdend met alle wensen met betrekking tot pati\u00ebntparticipatie kan deze opgezet worden om samenwerking te bevorderen van mensen met DMT2, zorgprofessionals, technologieontwikkelaars, beleidsmakers, andere belanghebbenden en onderzoekers, vanaf de ontwikkeling tot aan de toepassing en succesvolle implementatie van e-health. Dit artikel werd eerder gepubliceerd in TSG Tijdschrift Gezondheidswet (2021) 99:110-119 Volksgezondheidenzorg.info. Ranglijst aandoeningen op basis van voorkomen. https://www.volksgezondheiden zorg.info/ranglijst/ranglijst-aandoeningen-op-basis-vanv% C3%B3%C3%B3rkomen.Geraadpleegdop: 4sep2020. Sern\u00e9 E, Mourits P, Strien M van. Nationale Diabetes Registratie voor een toekomstbestendige en persoonsgerichte diabeteszorg in Nederland. Ned Tijdschr Diabetol. 2020;18:2. Rijksinstituut voor Volksgezondheid en Milieu. Volksgezondheid Toekomst Verkenning 2018: een gezond vooruitzicht. Bilthoven: RIVM;2018. Ministerie van Volksgezondheid, Welzijn en Sport. Nationaal Preventieakkoord, naar een gezonder Nederland. Den Haag: Ministerie van Volksgezondheid, Welzijn en Sport; 2018. 5. Raad voor de Volksgezondheid en Zorg. Zorg voor je gezondheid! Gedrag en gezondheid: de nieuwe ordening. Den Haag: RaadvoordeVolksgezondheidenZorg;2010. https://www.nictiz.nl/wpcontent/ uploads/E-health-Wat-is-dat.pdf. Geraadpleegd op: 27maart2020. Nictiz. E-Health, wat is dat? https://www.nivel.nl/nl/publicatie/samen-aan-zet-ehealth-monitor-2019. Geraadpleegd op: 10nov2020. Wouters M, Huygens M, Voogdt H, et al. Samen aan zet! eHealth-monitor2019. Chen L,Pei JH ,KuangJ, et al. Effect of lifestyle intervention in patients with type2diabetes: ameta-analysis. Metabolism. 2015;64:338-47. Idris I, HamptonJ, MoncrieffF,etal. Effectiveness of a digital lifestyle change program in obese and type 2 diabetes populations: service evaluation of real-world data. JMIR Diabetes. 2020;5:e15189. Little P, Stuart B, Hobbs FR, et al. An internet-based intervention with brief nurse support to manage obesity in primary care (POWeR+): a pragmatic, parallel-group, randomised controlled trial. Lancet Diab Endocrinol. 2016;4:821-8. Greenwood DA, Gee PM, Fatkin KJ, et al. Asystematicreview of reviews evaluating technology-enabled diabetes selfmanagement education and support. J Diab Sci Technol. 2017;11:1015-27. Ross J, Stevenson F, Lau R,et al. Factors that influence the implementation of e-health: asystematicreviewof systematic reviews (an update). ImplementSci. 2016;11:146.Alvarado MM, Kum HC, Gonzalez Coronado K, et al. Barriers to remote health interventions for type 2 diabetes: a systematic review and proposed classification scheme. JMedInternetRes. 2017;19:e28. Rogers EM.Diffusion of Innovations. NewYork: FreePress; 2003.Bruinessen IR van, Weel-Baumgarten EM van, Snippe HW, et al. Active patient participation in the development of an online intervention. JMIRResProtoc. 2014;3:e59. Gemert-Pijnen JEWC van, Nijland N, LimburgMvan, et al. A holistic framework to improve the uptake and impact of ehealth technologies. JMedInternetRes. 2011;13:e111. https://participatiekompas.nl/handboek-om-samenmet-patienten-de-zorg-te-verbeteren. Geraadpleegd op: 4sep2020. Raats I, Brink R van den, Wit F de. Handboek pati\u00ebnten- en cli\u00ebntenparticipatie. Verbeteren van de zorg samen met pati\u00ebnten/cli\u00ebnten. CBO, Provinciaal Pati\u00ebnten/Consumenten Platform Utrecht, Nederlandse Pati\u00ebnten Consumenten Federatie (NPCF). Bovenkamp H van de, Grit K, Bal R. Inventarisatie pati\u00ebntenparticipatie in onderzoek, kwaliteit en beleid. Rotterdam: Instituut Beleid en Management Gezondheidszorg ErasmusMC; 2008. Broeder L den, Lemmens L, Uysal S, et al. Public citizen science; perceived impacts on citizen scientists: a case study in low-income neighbourhoods in the Netherlands. CitizSciTheoryPract. 2017;2:1-17. Arnstein SR. Ladder of citizen participation. J Am Inst Planners. 1969;35:216-24. Lemmens LC, Bruin SR de, Struijs JN, et al. Patient involvement in diabetes care: experiences in nine diabetes caregroups. IntJ IntegrCare. 2015;15:e44.Beighton C, Victor C, Carey IM, et al. 'I'm sure we made it a better study . . . ': experiences of adults with intellectual disabilities and parent carers of patient and public involvement in a health research study. J Intellect Disab. 2019;23:78-96.Geppert C, Candilis P, Baker S, et al. Motivations of patients with diabetes to participate in research. AJOB EmpirBioeth. 2014;5(4):14-21.Barrett R, Levickis P, Naughton G, et al. Why families choose not to participate in research: feedback from nonresponders. JPeadiatrChildHealth. 2013;49(1):57-62. Beek A van. Zelfeffectiviteit. Ned Tijdschr Diabetol. 2015;13:26.Ujcic-Voortman JK, Schram MT. Jacobs-van der Bruggen MA, et al. Diabetes prevalence and risk factors among ethnicminorities. Eur JPublicHealth. 2009;19:511-5. Davis RE, Jacklin R, Sevdalis N, et al. Patient involvement in patient safety: what factors influencepatient participation and engagement? HealthExpect. 2007;10(3):259-67."} +{"text": "This month\u2019s theme is linked to the Prince Mahidol Award Conference on the intersection of health with climate change, environment and biodiversity. In the editorial section, Viroj Tangcharoensathien et al. (82) discuss the political commitments needed to mitigate the health impact of the climate crisis. Sarah Whitmee et al. (83) introduce a new database that tracks global carbon emissions. Fid Thompson (86\u201387) reports on urban planning changes needed to address climate change, flooding and sanitation. Joy Shumake-Guillemot tells Gary Humphreys (88\u201389) about recent climate and health initiatives in the World Meterological Organization.Sera L Young et al. (90\u2013101) investigate associations between food and water insecurities.Swetha Manohar et al. (140\u2013148) describe how healthy livelihoods depend on the Mekong River.Zlatko Nikoloski et al. (111\u2013120) measure adherence to public health and social measures. Adithya Pradyumna et al. (149\u2013151) examine mitigation measures in terms of climate justice.Nandita Saikia et al. (102\u2013110) study factors associated with registration coverage.Nicole Egerstrom et al. (130\u2013139) estimate lives and resources saved if guidelines were met.Mary C Sheehan (121\u2013129) analyses climate adaptation plans of large cities.Andrea Monica D Ortiz et al. (152\u2013154) list requirements for cyclone recovery.Laurie Laybourn-Langton (155\u2013157) calls for justice in climate negotiations. Jen Iris Allan (158\u2013160) describes the trade-offs in linking health to climate change advocacy."} +{"text": "Following publication of the original article (Hao et al. The original article (Hao et al."} +{"text": "In Nederland worden naar schatting 185.000 pati\u00ebnten met diabetes mellitus behandeld in de tweede of de derde lijn,2 een aantal dat naar verwachting toe zal nemen als gevolg van overgewicht en vergrijzing. Het exacte aantal polikinisch behandelde diabetespati\u00ebnten is echter onbekend. Ook is weinig bekend over de kenmerken en vooruitzichten van deze pati\u00ebnten. Om de kwaliteit van diabeteszorg in de toekomst te garanderen, is meer kennis nodig over deze groep diabetespati\u00ebnten. Het primaire doel van DPARD is om op lokaal, regionaal en landelijk niveau beter inzicht te krijgen in alle aspecten van de chronische poliklinische diabeteszorg, teneinde de kwaliteit van diabeteszorg op elk van deze niveaus verder te verbeteren. In 2017 werd op Wereld Diabetes Dag DPARD (Dutch Pediatric and Adult Registry of Diabetes) opgericht, de nationale registratie voor diabetespati\u00ebnten behandeld in de tweede en derde lijn.DPARD is opgericht door Stichting BIDON (Basisstructuur Innovatief Diabetes Onderzoek Nederland), een landelijk consortium van internisten, kinderartsen en diabetespati\u00ebnten. Sinds 2018 valt DPARD onder de vleugels van DICA, een stichting die 22 landelijke kwaliteitsregistraties beheert, met als doel de kwaliteit van zorg inzichtelijk te maken. Het bestuur van DPARD is de Clinical Audit Board (CAB), bestaande uit de bestuursleden van Stichting BIDON. In de wetenschappelijke commissie nemen afgevaardigden deel van centra die deelnemen aan DPARD. De CAB is in samenspraak met de wetenschappelijke commissie verantwoordelijk voor het vaststellen van de jaarlijks aan te leveren landelijke kwaliteitsinformatie, de doorontwikkeling van de registratie en bewaking van de datakwaliteit.In DPARD worden gegevens geregistreerd die betrekking hebben op pati\u00ebntkarateristieken, diagnostiek, behandeling, uitkomsten en complicaties. Alle typen diabetes mellitus worden ge\u00efncludeerd, behoudens zwangerschapsdiabetes, aangezien dit in principe een tijdelijke aandoening is. Data worden direct vanuit het elektronisch pati\u00ebntendossier (EPD) verzameld. Inclusie vindt hierbij plaats op basis van DBC- (diagnose-behandelcombinatie)declaratiecodes. Diabetesdiagnose, classificatie, complicaties en comorbiditeiten worden vastgelegd met ICD-10-codes, wat aansluit bij de manier waarop gegevens op landelijk niveau in Nederland in elk EPD vastgelegd worden. Data vanuit het EPD wordt gebundeld in een groot databestand. Dit gebeurt veelal door medewerkers van ICT- of EPD-afdelingen van zorginstellingen. Het bestand wordt verstuurd naar databewerker Medical Research Data Management (MRDM), waar de data wordt bewerkt, versleuteld en opgeslagen. In lijn met de huidige privacywetgeving zijn de gegevens niet meer herleidbaar tot de individuele pati\u00ebnt.3 Deze stap heeft gezorgd voor prioritering binnen ziekenhuizen en voor landelijke bekendheid. Het effect hiervan is evident zichtbaar in de inclusiecijfers. Ook de impact van de COVID-19-pandemie is duidelijk zichtbaar door de stagnatie van inclusie van pati\u00ebnten in 2020-2021.De afgelopen jaren zijn diverse fasen doorlopen om DPARD door te ontwikkelen tot een volwaardige kwaliteitsregistratie. Sinds de initiatiefase is er een stevig fundament gebouwd in de datastructuur van de registratie, de verzameling van data en de terugkoppeling aan ziekenhuizen in online dashboards. Vijf jaar na oprichting zijn er 48.152 pati\u00ebnten ge\u00efncludeerd in DPARD, afkomstig uit 29 medische centra in Nederland . Het ove4,5 Tot slot sluit DPARD sinds 2022 volledig aan op de jaarlijks verplicht aan te leveren landelijke kwaliteitsinformatie (diabetesindicatoren), waardoor het mogelijk is om al deze indicatoren eenvoudig via DPARD aan te leveren.Wat verder heeft bijgedragen aan de opkomst van DPARD is de samenwerking die is opgezocht met de twee grootste EPD-leveranciers van Nederland, om de registratie te integreren in het zorgproces en registratie aan de bron te vergemakkelijken. Uitgangspunt is dat DPARD het zorgproces vastlegt zoals het geregistreerd wordt in het EPD. Daarom wordt binnen het dagelijkse team achter DPARD veel aandacht besteed aan het optimaliseren van de aansluiting op het zorgproces en de verschillende EPD's teneinde registratielast te beperken. Dit is van belang omdat de prevalentie van pati\u00ebnten met diabetes mellitus in Nederland naar verwachting de komende jaren verder zal blijven stijgen.patient reported outcome measures (PROMs) binnen DPARD, conform de basisset diabetes zoals geformuleerd door de International Consortium for Health Outcomes Measurement (ICHOM).6 Om inzicht te krijgen in uitkomsten van zorg en praktijkvariatie zijn de door pati\u00ebnten gerapporteerde uitkomstmaten van belang om vast te stellen of pati\u00ebnten ook daadwerkelijk baat hebben bij een behandeling.7 Op deze manier kunnen de uitkomsten ook ingezet worden om samen met de pati\u00ebnt te beslissen in de spreekkamer, en kan DPARD duurzaam bijdragen aan verbetering van de kwaliteit van zorg voor de pati\u00ebnt met diabetes.Het hoofddoel van kwaliteitsregistraties is om het zorgproces inzichtelijk te maken met behulp van kwaliteitsinformatie, deze te evalueren en vervolgens in te zetten voor continue verbetering van de geleverde zorg. Er wordt toegewerkt naar het volledig doorlopen van deze verbetercyclus. Ondanks het recente eerste lustrum is DPARD nog steeds een registratie in ontwikkeling. Het bereiken van landelijke dekking voor DPARD binnen de interne geneeskunde en de kindergeneeskunde is op dit moment een prioriteit, zodat de pati\u00ebntkenmerken, uitkomsten en praktijkvariatie tussen ziekenhuizen en regio's van de Nederlandse diabeteszorg inzichtelijk worden. Ook is het essentieel dat de verzamelde gegevens valide en betrouwbaar zijn en blijven. Om deze reden is er een dataverificatietraject gestart, waarbij de inclusie van pati\u00ebnten en enkele belangrijke klinische variabelen geverifieerd worden in deelnemende centra met behulp van administratieve ziekenhuisdata. Om in de toekomst gedegen uitspraken te kunnen doen over de kwaliteit van zorg, is correctie voor pati\u00ebntgebonden karakteristieken ook van belang, de zogeheten casemixcorrectie. Ten slotte wordt in 2023 een start gemaakt met het integreren van joint effort van veel partijen. Wij danken de leden van de Clinical Audit Board van DPARD, de Wetenschappelijke Commissie en alle betrokkenen binnen the Dutch Institute for Clinical Auditing (DICA) voor hun inzet. Wij danken de Diabetes Vereniging Nederland, de Nederlandse Internisten Vereniging, de Nederlandse Vereniging van Kindergeneeskunde en de Nederlandse Diabetes Federatie voor hun steun. Tot slot danken wij alle participerende centra voor hun inspanningen.DPARD is bij uitstek een Bak JCG, Mul D, Sern\u00e9 EH, de Valk HW, Sas TCJ, Geelhoed-Duijvestijn PH, et al. DPARD: rationale, design and initial results from the Dutch national diabetes registry. BMC Endocr Disord. 2021 Dec;21(1). https://nivel.nl/nlNivel | Kennis voor betere zorg. Nederlands Instituut voor Onderzoek van de Gezondheidszorg [Internet]. Available from: https://www.internisten.nl/over-de-niv/magazine/de Vries S, Bak J, Verheugt C. Diabeteszorg naar een hoger niveau. Interne Geneeskd Mag voor internist [Internet]. 2021;3(12):18-20. Available from: Sun H, Saeedi P, Karuranga S, Pinkepank M, Ogurtsova K, Duncan BB, et al. IDF Diabetes Atlas: Global, regional and country-level diabetes prevalence estimates for 2021 and projections for 2045. Diabetes Res Clin Pract. 2022;183:109119. Nielen M, Poos R, Korevaar J. Diabetes mellitus in Nederland. Prevalentie en incidentie: heden, verleden en toekomst. Utrecht; 2020. Nano J, Carinci F, Okunade O, Whittaker S, Walbaum M, Barnard-Kelly K, on behalf of the DWG of the IC for HOM (ICHOM). A standard set of person-centred outcomes for diabetes mellitus results of an international and unified approach. Diabet Med. 2020;37(12):1959-2172. Lamberts MP, Drenth JPH, Van Laarhoven CJHM, Westert GP. Uitkomst van behandeling volgens pati\u00ebnten: Instrument om variaties in klinisch handelen terug te dringen. Ned Tijdschr Geneeskd. 2013;157(11):1-4.HighlightsLandelijke diabetesregistratie DPARD is eind 2017 opgericht en bestaat inmiddels vijf jaar.DPARD is een voorbeeld van een multidisciplinaire registratie, waarin poliklinisch behandelde kinderen en volwassenen met diabetes mellitus vervolgd worden over de tijd.De pati\u00ebntenvereniging en wetenschappelijke verenigingen zijn actief betrokken en onmisbaar in de ontwikkeling van de registratie. Inmiddels levert 40% van de Nederlandse centra gegevens aan en is 86% aangemeld. Aangeleverde kwaliteitsgegevens zijn inzichtelijk in interactieve dashboards en kunnen vergeleken worden met andere deelnemende instellingen. Directe extractie van gegevens uit het EPD helpt om registratielast te beperken.Het hergebruiken van data kan verder worden geoptimaliseerd als er aandacht blijft bestaan voor het gestructureerd vastleggen van gegevens in EPD's."} +{"text": "This month\u2019s theme issue is rehabilitation. In editorials, Director-General Tedros Adhanom Ghebreyesus (654) explains how the World Health Organization supports its member states in the provision of rehabilitation services. Alarcos Cieza et al. (655) make the case for change in the way these services are organized, staffed and funded. Fid Thompson (658\u2013659) reports on how Ukraine is building on rehabilitation work initiated by the government before the war. Abderrazak Hajjioui talks to Tatum Anderson (660\u2013661) about the need to prioritize multidisciplinary rehabilitation education, practice and research.Rajani Mullerpatan et al. (662\u2013668) pilot sustainable services in five villages.Limor Lavie & Tali Bar-Moshe (739\u2013743) examine lessons learnt after increased subsidies for hearing aids.Alessandra Da Ros et al. (669\u2013675) track referrals to a family and community physiotherapist.Kaori Yamaguchi et al. (699\u2013708) explore uses of insurance data in managing rehabilitation services.Thandi Conradie et al. (689\u2013698) compare national treatment guidelines.Didier Demey et al. (733\u2013738) link four Vietnamese universities with international providers of pre-service education.Simon D\u00e9cary et al. (676\u2013688) review the evidence for models of care.Tamara Chikhradze et al. (709\u2013716) indicate how to improve governments\u2019 purchasing of services.Tracey Smythe et al. (717\u2013725) examine the needs of children with congenital conditions.Cornelia Anne Barth et al. (726\u2013732) describe strategies to improve outcomes in low- and middle-income countries. Pete Skelton et al. (744\u2013746) look at what\u2019s required to improve emergency preparedness.Jim Campbell & Jody-Anne Mills (747\u2013748) outline workforce-related health systems and policy research."} +{"text": "In Noordermeer et al. (2002)"} +{"text": "The editorial team is grateful to all the experts who dedicated their time to peer-review our articles over the past 12 months. We wish to thank them and all those whose names are not listed below but gave helpful advice and support. We apologise to any reviewers whose names we may have missed.Our reviewers in 2022 were:Amos Adler; Rao Agam; Leonidas Alexakis; Sheikh Taslim Ali; Vanessa Allen; Erik Alm; Maria Joao Alves; Filipa Alves da Costa; F\u00e1tima Amaro; Maria an der Heiden; Aura Andreasen; Emmanuel Andr\u00e9; Andrea Antinori; Alberto Antonelli; Brett Archer; Tommi Asikainen; Tatjana Avsic-Zupanc; Benjamin R Baer; Henry C Baggett; Tamas Bakonyi; Eszter Balla; Susan Ballard; Mathieu Bangert; Cyril Barbezange; Ian Barr; Ulrike Baum; Edward Belongia; Alexandre Belot; Anna Beltrame; Shmuel Benenson; Beatrice Bercot; Matthijs Berends; Laura Berneking; Debra Bessen; Pierre Bessiere; Julie Bettinger; Simona Bignami; Jonas Bj\u00f6rk; Carina Blackmore; Lucille Blumberg; Benjamin Bl\u00fcmel; M\u00e1ir\u00edn Boland; Idesbald Boone; Maria Laura Boschiroli; Herve Bourhy; Amanda Bradley-Stewart; Michael Brandl; Carina Brehony; Nathan J Brendish; Anne Brisabois; Sylvain Brisse; Basil Britto Xavier; Eeva Broberg; Kim Brolin; Caroline Br\u00f6jer; Laura Bubba; Alicia Budd; Michael Busch; Karina Butler; Leon Caly; Christine Campese; Rafael Cant\u00f3n; Heather Carleton; Yehuda Carmeli; Jean-sebastien Casalegno; Kelsie Cassell; Alessandro Cassini; Nadim Cassir; Francesco Castelli; Jesus Castilla; Vincent Cattoir; Dominique Caugant; Orlando Cenciarelli; Muge Cevik; David Champredon; Andr\u00e9 Charlett; Remi Charrel; Cindy Chiu De Vazquez; Alexandra Choi; Michal Chowers; Nadia Ciampa; Jose Miguel Cisneros; Hazel Clothier; Bruno Coignard; Vittoria Colizza; M\u00e9lanie Colomb-Cotinat; John Conly; Jeff Connell; Martin Cormican; Ana Paula Coutinho Rehse; Benjamin Cowie; Benjamin Cowling; Fortunato D'Ancona; Gavin Dabrera; Viktor Dahl; Ghassan Dbaibo; Brechje de Gier; Marit de Lange; Franck de Laval; Elisabeth Delarocque-Astagneau; Alessandra della Torre; Oliver Denis; Charlotte Deogan; Brecht Devleesschauwer; Nolwenn Dheilly; Sofie Dhollander; Santina Di Bella; Antonino Di Caro; Liselotte Diaz H\u00f6gberg; Frederika Dijkstra; Jac Dinnes; Gerhard Dobler; Margaret Doll; Lisa Domegan; Laurent Dortet; Jan Drexler; Christian Drosten; Julian Druce; Timothee Dub; Erwin Duizer; Jake Dunning; David Durrheim; Florence D\u00e9barre; Tim Eckmanns; Michael Edelstein; Androulla Efstratiou; Ehud Elnekave; Devy Emperador; Giulia Errico; Allahna Esber; Laura Espenhain; W. Douglas Evans; Daniel Faensen; James Fielding; Helen Fifer; Julie Figoni; Antonietta Filia; Jose Ramon Fiore; Robert Fischer; Brendan Flannery; \u202aDaniele Focosi; Arnaud Fontanet; Laure Fonteneau; Nicoletta Formenti; Ron Fouchier; Nelly Fournet; Christina Frank; Sebastian Funk; Emmanouil Galanakis; Andrea Gervelmaeyer; Sarah Gerver; Isaac Ghinai; Yvonne Gilleece; Gary Ginsberg; Aharona Glatman-Freedman; Daniel Golparian; Maria Gossens; C\u00e9line Gossner; Magnus Gottfredsson; Rok Grah; M. Lindsay Grayson; Margrethe Greve-Isdahl; Martin P Grobusch; Thomas Gronthal; Thomas Gr\u00f6nthal; barbara Gr\u00fctzmacher; Claire Guinat; Bernardo Rafael Guzman Herrador; Stephan G\u00f6ttig; Sebastian G\u00fcnther; Stefan Hagel; Alvin X Han; Sonja Hansen; Lisa Hansen; Kayla Hanson; Thomas Harder; Aaron Harris; David M. Hartley; Henrik Hasman; Margaret Haworth-Brockman; Florian Heger; Janneke Heijne; Ulrich Heininger; Franz Heinz; Victoria Hernando; Marta Hern\u00e1ndez-Garc\u00eda; Megan Hofmeister; Mark Holmes; Martin Holt; Camilla Holten; Ralph Huits; Ralph Huits; Anders Husby; Anders Hviid; Michael H\u00f6hle; Judith H\u00fcbschen; Derval Igoe; Victoria Indenbaum; John PA Ioannidis; Gregorio Iraola; Takahiro Itaya; Jesus I\u00f1igo; St\u00e9phanie Jacquinet; Agatha Jassem; Claire Jenkins; Silvia Jimenez-Jorge; Kari Johansen; Annette Jurke; Annette Jurke; Stefanie Kadykalo; G\u00fcnter Kampf; Tommi Karki; Taishi Kayano; M\u00e1ria Kazim\u00edrov\u00e1; Lee Kennedy-Shaffer; Gilbert Kersh; John Kinsman; Nishant Kishore; Esther Kissling; Masaaki Kitajima; Wioleta Kitowska; Lene Jung Kj\u00e6r; Lene Jung Kj\u00e6r; Sofieke Klamer; Irena Klavs; Petra Klepac; Anders Koch; Anke Kohlenberg; Britta Kohlmorgen; Archana Koirala; Shihoko Komine-Aizawa; Rolf Kramer; Marcela Krutova; Kevin Kuchinski; Anna Kuehne; EJ Kuijper; Michael Kundi; Sohvi K\u00e4\u00e4ri\u00e4inen; Martin K\u00e5berg; Csaba K\u00f6dm\u00f6n; Hannah K\u00f6nig; Raskit Lachmann; Nina Lagerqvist; Jeffrey Lazarus; St\u00e9phane Le Vu; Nelson Lee; Jean-Daniel Leli\u00e8vre; Vivian Leung; Kathy Leung; Troels Lillebaek; Bruno Lina; Anders Lindblom; Adrian Lison; Florence Lot; Claudia Lucarelli; \u00c5ke Lundkvist; Yaniv Lustig; Irja Lutsar; Theodore Lytras; Outi Lyytik\u00e4inen; Joaqu\u00edn L\u00f3pez-Contreras Gonzalez; Emma L\u00f6f; Renke L\u00fchken; Gon\u00e7alo Macedo; Raina MacIntyre; Guerrino Macori; Alexandra Mailles; John Mair-Jenkins; Anna Maisa; Gannon C.K. Mak; Jean-Michel Mansuy; Ulrich Marcus; Robby Markwart; Robby Markwart; Robby Markwart; Alexandra Marmor; Louise Marron; Katherine E. Marshall; Stacey W Martin; Iv\u00e1n Mart\u00ednez-Baz; S\u00e9bastien Matamoros; Wesley Mattheus; Sebastian Maurer-Stroh; Andrea McCollum; Scott McDonald; James McMahon; Jim McMenamin; Adam Meijer; Anika Meinen; Roberto Melano; Angeliki Melidou; Tanya Melillo Fenech; Stefano Merler; Kevin Messacar; Eline Meyers; Chris Ka Pun Mok; Jose Molina-Mora; Monica Monaco; Orna Mor; Jacob Moran-Gilad; Alain Moren; Leah Moriarty; Jo\u00ebl Mossong; Aikaterini Mougkou; Laurent Moulin; Antons Mozalevskis; Judith Mueller; Maximilian Muenchhoff; Michio Murakami; Benjamin Murrell; David Muscatello; Otilia M\u00e5rdh; K\u00e5re M\u00f8lbak; Tony Nelson; Sema Nickbakhsh; Andreas Nitsche; Harold Noel; Paulo Nogueira; Baltazar Nunes; Teresa Nygren; Sheila F. O'Brien; Joan O'Donnell; Sean O'Leary; Doris Oberle; Nicholas Ogden; Ajibola Omokanye; Joshua Osowicki; Juliette Paireau; Scott Pallett; Annalisa Pantosti; Anna Papa; John Papp; Dimitrios Paraskevis; Prabasaj Paul; Jean-Michel Pawlotsky; Lara Payne; Richard Pebody; Malik Peiris; Pasi Penttinen; Mariana Perez Duque; Carla Perrotta; Eskild Petersen; Patrizio Pezzotti; Germ\u00e1n Pe\u00f1alva; Martin Pfeffer; Anastasia Pharris; Anastasia Phillips; Nick Phin; Roan Pijnacker; Rosa Pint\u00f3; Denis Pi\u00e9rard; Diamantis Plachouras; Mario Plebani; Catherine Pl\u00fcss-Suard; Laurent Poirel; Piero Poletti; Stefano Pongolini; Leo Poon; Stephanie Popping; Bastian Prasse; Elisabeth Presterl; Henrieke Prins; Katarina Prosenc; Mirja Puolakkainen; Tina Purnat; Mario Ramirez; Ruwan Ratnayake; Giovanni Ravasi; Cameron Razieh; Gili Regev-Yochay; Hariharan Regunath; Ralf Reintjes; Jordi Rello; Ute Rexroth; Giovanni Rezza; Flavia Riccardo; Maria Luisa Ricci; Matteo Ricc\u00f2; Wendy Rice; Maximilian Riess; Thomas V. Riley; Caterina Rizzo; Ilia Rochlin; Nash Rochman; Ana Rodrigues; Ana Rodrigues; Jes\u00fas Rodr\u00edguez-Ba\u00f1o; Maria Romay-Barja; Dan Romer; Angela Rose; John Rossen; Gian Maria Rossolini; Maja Rupnik; Jan Rupp; Kati R\u00e4is\u00e4nen; Raquel Sabino; \u00d6rjan Samuelsen; Frank Sandmann; Jussi Sane; Andr\u00e9 Scherag; Gabrielle Schittecatte; Axel Schmidt; Madlen Schranz; Luigi Sedda; Philippe Selhorst; Luca Settanni; Azam Ali Sher; George Shirreff; Rami Sommerstein; Gianfranco Spiteri; Jean-Paul Stahl; John Stelling; Ellen Stobberingh; Franc Strle; Lorenzo Subissi; Jonathan Suk; Sheena Sullivan; Gang Sun; J\u00e1nos S\u00e1ndor; Emi Takashita; Heather Tate; Lara Tavoschi; Kjetil Elias Telle; Bernd-Alois Tenhagen; Karen Terio; Heidi Theeten; Craig Thompson; Valtyr Thors; Laura Tomassone; Sara Tomczyk; Ana Torres; Maria Elena Tosti; Alenka Trop Skaza; Sarah Tschudin-Sutter; Gro Tunheim; Yuki Uehara; Soren Uldum; Veronika U\u010dakar; Nirma Vadlamudi; Chris Van Beneden; Wim Van Bortel; Michiel van Boven; Wim van der Hoek; Mark van der Linden; Marianne van der Sande; Sebastiaan van Hal; Jasper Van Heuverswyn; Angela van Hoek; Esther Van Kleef; Maaike van Mourik; Sonja van Roeden; Carmen Varela Martinez; Aisling Vaughan; Tomas Vega-Alonso; Pierre Verger; Sophie von Dobschuetz; Rengina Vorou; Jacco Wallinga; Jan Walter; Stella Watson; Guido Werner; David Whiley; Robert Whittaker; Micael Widerstr\u00f6m; Andreas Widmer; Annelies Wilder-Smith; Hendrik Wilking; Thomas Williams; Christopher Williams; Carl-Heinz Wirsing von K\u00f6nig; Sook San Wong; Jessica YT Wong; Ian Woolley; Tom Woudenberg; Cameron Wright; Stanley Xu; Barbara Yawn; Dana Yelin; Karene Yeung; Philipp Zanger; Peter Zarb; Petr Zeman; Dominik Zenner; Ye Zhang; Shi Zhao; Ruth Zimmermann; Kate Zinszer; Ljiljana Zmak; Julio \u00c1lvarez S\u00e1nchez.\u202c"} +{"text": "Dit artikel beschrijft het ontstaan, de verschillende onderdelen en de thema\u2019s van de LSM, en de taken en rollen van de samenwerkende partijen. Daarnaast beschrijft het hoe de gegevens verzameld worden en waar de gegevensverzamelingen op te vragen zijn voor onderzoek. Tot slot bevat het enkele voorbeelden van cijfers en trends over de periode 2014\u20132020 ter ondersteuning van gezondheidsbeleid Voor de onderbouwing van nationaal gezondheidsbeleid is een monitoringssysteem van belang waarin cijfers over leefstijl en gezondheid worden gemonitord over tijd. Deze cijfers worden vaak verzameld via vragenlijstonderzoek in een steekproef van de bevolking. In Nederland kennen we een geschiedenis van diverse vragenlijstonderzoeken die zich vaak richtten op \u00e9\u00e9n onderwerp en worden uitgevoerd door thema-instituten, zoals het Continu Onderzoek Rookgewoonten (COR) van Stivoro/het Trimbos-instituut. Daarnaast waren er onderzoeken die zich richtten op meerdere onderwerpen, waaronder leefstijlgedragingen, zoals de Gezondheidsenqu\u00eate (GE) van het Centraal Bureau voor de Statistiek (CBS) .In 2011 startte het ministerie van Volksgezondheid, Welzijn en Sport (VWS) met de herijking van het leefstijlbeleid. Een van de vragen die daaruit volgde was of de verschillende gegevensverzamelingen over leefstijl effici\u00ebnter en in samenhang met elkaar ingericht konden worden . In 2012In 2013 en 2014 werd dit voorstel uitgewerkt tot een vernieuwde opzet van de verschillende monitoringsactiviteiten onder de noemer \u2018de Leefstijlmonitor\u2019, ondersteund door het ministerie van VWS. Het doel van de Leefstijlmonitor (LSM) is het effici\u00ebnter en meer samenhangend organiseren van de landelijke gegevensverzameling op het gebied van leefstijl in Nederland [De LSM is een product van de samenwerking tussen partijen die zich richten op leefstijl. De volgende partijen vormen samen het Consortium LSM: Trimbos-instituut, Rutgers, Soa Aids Nederland, Pharos, VeiligheidNL, Voedingscentrum, CBS en GGD GHOR Nederland. Het RIVM fungeert binnen de LSM als co\u00f6rdinator van 1)\u00a0de LSM, 2)\u00a0het netwerk kernindicatoren sport en bewegen en 3)\u00a0de VoedselConsumptiePeiling (VCP). De genoemde partijen zijn samen verantwoordelijk voor de inhoud van de LSM en publiceren cijfers voor gezondheidsbeleid op basis van de gegevensverzamelingen van de LSM. De taken van de partijen en hun rollen binnen de LSM staan beschreven in tab.\u00a0roken;alcoholgebruik;drugsgebruik (inclusief sportprestatieverhogende middelen);seksuele gezondheid;bewegen, sport en zitten;voeding;gezond gewicht;ongevallen (inclusief sportblessures).De thema\u2019s die onder de LSM vallen betreffen de thema\u2019s waarop de thema-instituten zich richten en waarvan een relatie met gezondheid is vastgesteld , 4, namewww.leefstijlmonitor.nl [De LSM maakt gebruik van verschillende gegevensverzamelingen sinds 2014. Binnen de LSM zijn op basis van methodologische overwegingen afspraken gemaakt over welke gegevensverzameling de voorkeur heeft voor het vergaren van cijfers voor gezondheidsbeleid binnen een bepaald thema (\u2018preferente gegevensverzameling\u2019). Tabel\u00a0nitor.nl .BronLeefKern van de Leefstijlmonitor (LSM-K)de Gezondheidsenqu\u00eate (GE);het Peilstationsonderzoek Scholieren (PEIL);de Health Behaviour in School-aged Children study (HBSC-onderzoek).Onder de LSM\u2011K vallen drie gegevensverzamelingen met een vaste basis aan vragen. Dit zijn:De gegevensverzamelingen worden geschaard onder twee onderdelen:N\u202f~\u20099.500). Er wordt een random steekproef van mensen uit particuliere huishoudens getrokken uit de Basisregistratie Personen (BRP). Personen worden in eerste instantie uitgenodigd de vragenlijst via het internet in te vullen. Als ze niet via internet reageren, worden ze opnieuw benaderd via huis-aan-huisbezoeken met het verzoek de vragen uit de vragenlijst alsnog tijdens een persoonlijk interview te beantwoorden. Sinds 2018 wordt slechts een deel van de non-respondenten op de internetvragenlijst opnieuw benaderd voor een persoonlijk interview. Deze interviews worden vooral ingezet bij bepaalde groepen mensen die slecht responderen via internet. Deze zogenaamde doelgroepgerichte benadering verbetert de representativiteit [De GE wordt jaarlijks door het CBS uitgevoerd onder personen van 0\u00a0jaar en ouder . Kinderen uit het basisonderwijs worden niet meegenomen voor de LSM. Er is bij de opzet van de LSM gekozen voor het gebruik van gegevens over deze thema\u2019s op basis van PEIL en HBSC in plaats van de GE voor jeugd, omdat deze thema\u2019s in het bijzonder van belang zijn voor de jeugd en PEIL en HBSC een grotere groep jongeren bevragen dan de GE. Doordat meer dan 90% van de jongeren van de geselecteerde klassen deelnemen aan het onderzoek worden moeilijk bereikbare groepen beter bereikt.2.Aanvullende modules van de Leefstijlmonitor (LSM\u2011A)LSM\u2011A Bewegen en Ongevallen;LSM\u2011A Middelen;LSM\u2011A Seksuele Gezondheid in Nederland;Seks onder je 25e ;PEIL;VCP.Onder de LSM\u2011A vallen gegevensverzamelingen die aanvullende en verdiepende informatie binnen een bepaald thema bieden. Deze gegevensverzamelingen zijn:Voor roken, alcohol, drugs en seksuele gezondheid onder de jeugd worden gegevens uit PEIL en HBSC gebruikt. Beide onderzoeken worden om de vier jaar uitgevoerd door de Universiteit Utrecht, SCP en het Trimbos-instituut. De vraagstellingen van de thema\u2019s die gebruikt worden voor de LSM worden tussen de beide gegevensverzamelingen afgestemd, waarmee ze voor tweejaarlijkse cijfers zorgen. In PEIL en HBSC wordt een steekproef van scholen in het regulier voortgezet onderwijs getrokken om per deelnemende school via een selectie van klassen leerlingen uit alle leerjaren van het vmbo, havo en vwo te includeren. Voor de cijfers in de LSM wordt alleen gerapporteerd over jongeren van twaalf tot en met zestien jaar (N\u202f~\u200910.000). Voor deze gegevensverzamelingen wordt een vaste set vragen gesteld om trends te kunnen volgen die een aanvulling vormen op de LSM\u2011K. Daarnaast is er ruimte voor een veranderende set vragen waarmee ingespeeld kan worden op een veranderende informatiebehoefte vanuit gezondheidsbeleid en actualiteiten. Zo wordt in de LMS\u2011A Bewegen en Ongevallen naast beweeg- en sportgedrag ook zitgedrag uitgevraagd en worden meer vragen gesteld over de context waarin wordt bewogen en ongevallen zijn voorgekomen. In de LSM\u2011A Middelen wordt behalve naar roken, alcohol en drugs ook naar sportprestatieverhogende middelen (ofwel doping) gevraagd. De verdiepende vragen gaan bijvoorbeeld over problematisch alcoholgebruik en kennis van gezondheidsadviezen en de LSM\u2011A Seks onder je 25e (N\u202f~\u200920.000 18- tot 24-jarigen) zijn vanaf 2017 met elkaar ge\u00efntegreerd, omdat de doelgroepen van beide onderzoeken deels overlappen. Dat wil zeggen dat de vragenlijsten van de twee onderzoeken vergelijkbaar zijn gemaakt en de gegevens in dezelfde periode zijn verzameld. De vragenlijst dekt een breed scala van aan seksualiteit gerelateerde onderwerpen, zoals seksueel gedrag, risico op soa\u2019s en onbedoelde zwangerschap en seksueel geweld. Bij beide onderzoeken is gebruikgemaakt van een random steekproef uit de BRP. Bij de LSM\u2011A Seks onder je 25e zijn ook jongeren via scholen benaderd. De BRP-steekproefpersonen zijn uitgenodigd om via internet deel te nemen. Personen die niet via internet reageren en boven de 25\u00a0jaar zijn kregen een papieren vragenlijst thuis gestuurd. In 2022 worden deze gegevensverzamelingen herhaald.De LSM\u2011A Seksuele gezondheid in Nederland (Bij PEIL (zie ook LSM-K) worden naast basisvragen om prevalentiecijfers van roken, alcohol- en druggebruik en veilige seks te verkrijgen, ook verdiepende vragen gesteld. Deze vragen geven informatie over bijvoorbeeld de manier waarop de jeugd aan rook- en alcoholproducten komt of over hun attitude over middelengebruik.N\u202f~\u20094.000) woonachtig in Nederland. Bij hen is de consumptie van voedingsmiddelen nagevraagd met behulp van een computergeassisteerde 24-uursvoedingsnavraagmethode, waarin heel specifiek is nagevraagd welke producten er zijn geconsumeerd en hoeveel.De VCP geeft inzicht in de consumptie van voedingsmiddelen, de inname van macro- en microvoedingsstoffen en de inname van potentieel schadelijke chemische stoffen, en in de ontwikkelingen hiervan (trends), en informatie over de milieudruk. De VCP 2012\u20132016 heeft gebruikgemaakt van een consumentenpanel van 1\u2011 tot 79-jarigen (De VCP en de GE vullen elkaar aan op het gebied van voeding. De VCP brengt gedetailleerd in kaart (op voedingsmiddelen- en nutri\u00ebntenniveau) wat Nederland eet en drinkt. De GE geeft enkel inzicht in een aantal voedingsmiddelen aan de hand van algemene vragen. De VCP brengt in zijn algemeenheid de voeding beter in beeld, maar wordt bij minder mensen \u00e9n minder vaak uitgevoerd dan de GE. Daarom is het op basis van de GE makkelijker uitspraken te doen over trends en verschillen tussen specifieke groepen. Door de verschillen in de methodiek zijn de cijfers over consumptie van groente en fruit echter hoger op basis van de GE dan op grond van de VCP. Bij de duiding van deze cijfers wordt hier rekening mee gehouden.Om cijfers te verkrijgen die representatief zijn voor de Nederlandse bevolking worden de prevalentiecijfers bij alle gegevensverzamelingen van de LSM gecorrigeerd met een weegfactor . De weegEen aantal gegevensverzamelingen binnen de LSM is geharmoniseerd met lokale en/of internationale gegevensverzamelingen. De vragen over roken en alcohol (met uitzondering van 2016), lengte, gewicht en bewegen in de GE zijn geharmoniseerd met de vragen over deze onderwerpen in de Gezondheidsmonitor (GM) Volwassenen en Ouderen van de GGD\u2019en. Deze verzamelt eens in de vier jaar gegevens op regionaal en lokaal niveau ter ondersteuning van lokaal, regionaal en landelijk gezondheidsbeleid . Door deDe GE wordt ook gebruikt in een internationale context. De European Health Interview Survey (EHIS) wordt eenmaal in de zes jaar uitgevoerd in opdracht van de Europese Commissie. De GE levert cijfers aan op basis van de bestaande vragen of met een aantal aanpassingen om aan de eisen van de EHIS te voldoen .Het HBSC-onderzoek is ook een bron die in een internationale context wordt gebruikt . Het HBSwww.leefstijlmonitor.nl, www.wateetnederland.nl, www.sportenbewegenincijfers.nl en www.nationaledrugsmonitor.nl. Daarnaast zijn de cijfers terug te vinden op de websites van de verschillende consortiumpartners en op websites over de staat van de volksgezondheid en zorg in Nederland van het ministerie van VWS, www.volksgezondheidenzorg.info en www.staatvenz.nl. Deze webpagina\u2019s worden overigens ook geregeld gebruikt door personen die in de praktijk op het gebied van leefstijl werken, zoals fysiotherapiepraktijken die willen weten welke doelgroepen achterblijven in beweeggedrag, zodat ze hun interventies daar op kunnen richten.De cijfers die uit de kern- en aanvullende modules van de LSM komen dienen in de eerste plaats ter ondersteuning van het landelijk gezondheidsbeleid en worden in verschillende producten voor het ministerie van VWS gepubliceerd. Dit betreft rapportages, zoals de Nationale Drug Monitor , Letselswww.leefstijlmonitor.nl is te vinden bij welke partij en onder welke voorwaarden de gegevensverzamelingen op te vragen zijn. Een aantal gegevensverzamelingen is beschikbaar via het CBS . Daardoor zijn de gegevensverzamelingen te koppelen aan andere databronnen bij het CBS, zoals registraties met betrekking tot medicatie en zorg, uitkeringen en woon- of buurtkenmerken. Dit biedt extra mogelijkheden voor verdiepend onderzoek.Derden kunnen de gegevensverzamelingen of maatwerkanalyses van de LSM voor onderzoeksdoeleinden aanvragen. Op de webpagina \u2018Data aanvragen\u2019 van Hieronder volgt een aantal voorbeelden van cijfers en trends van 2014\u20132020 die specifiek worden gebruikt ter ondersteuning van het gezondheidsbeleid, zoals het monitoren van de doelstellingen in het Nationaal Preventieakkoord en het Sportakkoord , 14. De In het Nationaal Preventieakkoord zijn verschillende ambities uitgesproken over de thema\u2019s roken, overgewicht en alcohol, zoals \u2018In 2040 rookt minder dan 5% van de volwassenen >\u202f18\u00a0jaar)\u2019. Figuur\u00a08\u00a0jaar\u2019. Naast deze thema\u2019s is de voedselconsumptie een belangrijk onderdeel van het preventiebeleid ter voorkoming van chronische ziekten. Op basis van VCP/Leefstijlmonitor, RIVM 2012\u20132016, blijkt dat onder 1\u2011 tot 79-jarigen gemiddeld 131 gram groente en 113 gram fruit per dag wordt gegeten. Circa\u00a01 op de 6\u00a0van de volwassenen eet de door de Gezondheidsraad aanbevolen 200 gram of meer groente of fruit.Het aandeel personen dat voldoet aan de beweegrichtlijnen en dat wekelijks sport wordt onder andere gemonitord om de voortgang van de uitvoering van het Sportakkoord te bepalen. In fig.\u00a0Figuur\u00a0De cijfers laten zien dat het gebruik van cannabis en andere drugs tussen 2014 en 2020 in het voorgaande jaar bij volwassenen redelijk stabiel is gebleven (respectievelijk ongeveer 7% en ongeveer 4%).Het aandeel volwassenen dat risicovolle seks ) heeft gehad en zich heeft laten testen op seksueel overdraagbare aandoeningen is in diezelfde periode nagenoeg stabiel gebleven. De percentages liggen respectievelijk rond de 2\u00a0tot 3% en de 5\u00a0tot 6%.Het aantal letsels per persoon als gevolg van een priv\u00e9ongeval in het voorgaande jaar is tussen 2015 en 2020 ook nagenoeg stabiel gebleven en ligt rond de 2\u00a0tot 3\u00a0letsels.Het terugdringen van gezondheidsverschillen staat hoog op de agenda bij het ministerie van VWS . Binnen Een van de sterke punten van de LSM is dat verschillende instituten werkzaam op het gebied van leefstijl hun krachten en expertise hebben gebundeld om de samenhang tussen de gegevensverzamelingen te borgen. Zo worden in \u00e9\u00e9n vragenlijst (GE) alle leefstijlfactoren uitgevraagd, waardoor naar de onderliggende verbanden gekeken kan worden in samenhang met gezondheid. Daarnaast is een aantal gegevensverzamelingen beschikbaar bij het CBS, waardoor diepgaander onderzoek mogelijk is door gegevensverzamelingen van de LSM te koppelen aan andere databronnen. Deze onderzoeken bieden een schat aan informatie voor het vormen van gezondheidsbeleid. Zo is het monitoren van informatie waarbij leefstijl naar onderwijsniveau en inkomen wordt uitgesplitst bijvoorbeeld van belang bij het stellen van doelen binnen het Nationaal Preventieakkoord. Een ander pluspunt is dat binnen alle onderzoeken van de LSM wordt ingezet op het verhogen van de representativiteit van de steekproef. Dit wordt bijvoorbeeld gedaan door de uitnodigingen te richten op bepaalde doelgroepen die minder goed responderen (een doelgroepgerichte benadering).Naast sterke punten kent de LSM ook zwakkere punten. De gegevensverzamelingen van de LSM zijn cross-sectioneel. Dit is geschikt voor het doel waarvoor ze is opgezet, maar minder bruikbaar voor vragen over bijvoorbeeld veranderingen in leefstijl voorafgaand aan gezondheidsklachten. Daarnaast zijn de gegevens voornamelijk verzameld met vragenlijsten. Hoewel er wordt ge\u00efnvesteerd in het waarborgen van de representativiteit , blijft Ondanks de beperkingen hebben we dankzij de LSM een goede indicatie van de algemene gezondheidstrends en kunnen we verschillen tussen bepaalde doelgroepen toetsen. Bij gebruik van de cijfers moet hier echter rekening mee worden gehouden. Om de representativiteit verder te verbeteren onderzoekt het consortium de mogelijkheden om andere methoden toe te voegen om specifieke doelgroepen beter te kunnen includeren. Bij het afnemen van de vragenlijst door middel van persoonlijke interviews kunnen we denken aan het inzetten van interviewers die de culturele achtergrond van de respondent delen. Een andere mogelijkheid is het inzetten van meer kwalitatieve vormen van onderzoek bij specifieke doelgroepen, zoals personen die in probleemwijken wonen, scholieren van het middelbaar beroepsonderwijs (mbo) of mensen met een verstandelijke beperking. Het consortium bekijkt bij dergelijke opties in hoeverre ze in te passen zijn in bestaande monitoringsstructuren, of dat aanvullende onderzoeken nodig zijn.Het consortium werkt ook aan het verbeteren van methoden om leefstijl te monitoren. Zo wordt op nationaal en Europees niveau onderzocht of bewegen met een objectieve maat gemonitord kan worden en wordtDe LSM omvat een rijke verzameling aan gegevens van verschillende leefstijlfactoren. De LSM zorgt voor effici\u00ebntie en samenhang tussen verschillende gegevensverzamelingen dankzij de samenwerking tussen partijen op het gebied van leefstijl en gezondheid. De gegevens worden veelvuldig gebruikt ter ondersteuning van het gezondheidsbeleid, zoals het monitoren van de doelen binnen het Nationaal Preventieakkoord en het Sportakkoord. De gegevensverzamelingen zijn op te vragen voor onderzoeksdoeleinden en kunnen vaak gekoppeld worden aan andere databronnen, waarmee een grote verscheidenheid aan gezondheidsvraagstukken kan worden onderzocht."} +{"text": "In the editorial section, Ryan McBain et al. (358) introduce an initiative to track HIV resource allocation and costs. Jane Barratt (359) argues for specific measures to improve vaccination coverage in older adults.Gary Humphreys (362\u2013363) reports on continued resolve in the effort to eradicate polio and talks to John Nkengasong (364\u2013365) about the establishment of the Africa Centers for Disease Control.Tanya Wright et al. (385\u2013401) review the evidence for factors associated with burnout.Ali Abbas Rizvia & Abhishek Singh (375\u2013384) explore predictors of vaccine uptake in older adults.Mohamed Reda Bassiouny and Azza R Elhadidy (402\u2013408) describe foundations of an effective system.Miguel Angel Garcia-Bereguiain et al. (411\u2013412) argue for more stringent regulation of test kits. Tamim Alsuliman et al. (409\u2013410) make the case that formal guidelines are needed.Margaret McCarron et al. (366\u2013374) examine laboratory network performance in the context of reduced funding."} +{"text": "Additional article information.In Yang Yu et al.,"} +{"text": "Lucia (first name) Schiavon (last name)Alessandro (first name) Perini (last name)Giulia (first name) Brunello (last name)Giada (first name) Ferrante (last name)Massimo (first name) Del Fabbro (last name)Daniele (first name) Botticelli (last name)Fouad (first name) Khoury (last name)Stefano (first name) Sivolella (last name)Following publication of the original article , we have"} +{"text": "In this Special Issue, many original contributions concerning serological methods for SARS-CoV-2 were collected, some of them with implications about therapeutics.Bonifacio et al. demonstrVespa et al. used GSPPeter et al. from Germany confirmeDa Silva et al. from Luxembourg confirmeCia et al. from Belgium performeWang et al. from Taiwan providedFinally, we and colleagues from Johns Hopkins University systematically reviewed current knowledge on immune escape capabilities of SARS-CoV-2 against both anti-Spike mAbs and COVID-19 convalescent plasma (CCP) ["} +{"text": "In the editorial section Akhil Bansal (2) argues for equitable distribution of vaccines to prevent COVID-19. Kristen Meagher et al. (3) examine gender and leadership in the health-sector response to conflict-affected settings.Andrei Shukshin (6\u20137) reports on an initiative in the Russian Federation to improve food sources by the building of local greenhouses in remote communities. Rose Gana Fomban Leke talks to Gary Humphreys (8\u20139) about rethinking approaches to malaria elimination and her efforts to support the next generation of scientists in Cameroon.Luhua Zhao et al. (40\u201349) document data gaps.Phuong Hong Nguyen et al. (20\u201329) find unequal coverage for women and children.Jessica Craig et al. (50\u201359) compare national treatment guidelines.ANM Ehtesham Kabir et al. (10\u201319) study implementation barriers.Pierpaolo Di Carlo et al. (78\u201380) provide information in minority languages.Sanjay K Mohanty et al. (30\u201339) quantify missed opportunities.Anne Loarec et al. (60\u201369) pilot a mother-to-child transmission prevention programme.Arachu Castro et al. (81\u201383) advocate for measures to address climate change and social inequalities.Hannah Nissan et al. (70\u201377) describe climate-sensitive nutrition programmes."} +{"text": "In the editorial section, Vageesh Jain and Sam Tweed (322) argue that consensus is needed to standardise COVID-19 control objectives. Hussain Abbas Zaidi and Charles D Wells (323) examine the use of digital health technologies to improve adherence to tuberculosis treatment.In the news section, Lynn Eaton (326\u2013327) reports on initiatives designed to prevent the next pandemic. Helen Rees talks to Gary Humphreys (328\u2013329) about lessons learnt in pandemic prevention and the need to maintain a focus on polio eradication.Zujin Luo et al. (374\u2013380) document the process used to create a designated hospital.Dan N Tran et al. (388\u2013392) detail a revolving pharmacy model for essential medicines.Alexander Rosewell et al. (381\u2013387) pilot mobile and geographic information technologies.Nayawadee Kaweenuttayanon et al. (393\u2013397) describe the deployment of village health volunteers.Kerri Viney et al. (330\u2013341) examine a regional tuberculosis response framework.Catherine Machalaba et al. (342\u2013350) identify gaps in national pandemic preparedness plans.Amy Elizabeth Parry et al. (351\u2013358) examine what is needed to achieve collective competence.Rachel Vanderkruik et al. (359\u2013373) review the evidence for mental health outcomes.Alessandro Cassini et al. (398\u2013401) call for more research on the epidemiology of sepsis.Vanessa Brizuela et al. (402\u2013404) describe the need for local evidence in humanitarian settings."} +{"text": "In editorials this month, Viroj Tangcharoensathien et al. (238) call for papers for a special issue on the impact of climate change on biodiversity, agriculture and health. Lisa L Carter et al. (239) propose the next decade\u2019s global genomic surveillance strategy for pathogens with pandemic and epidemic potential. Tedros Adhanom Ghebreyesus et al. (240) present WHO recommendations for resilient health systems.Tatum Anderson (243\u2013244) reports on the environmental implications of medical waste generated by the COVID-19 response. Benjie Foscablo talks to Gary Humphreys (245\u2013246) about the pressures of nursing during the COVID-19 pandemic, why nurses are leaving the profession, and the urgent need to improve working conditions.Alice Nanelin Guingan\u00e9 et al. (256\u2013267) evaluate a screening programme for partners and children of pregnant women.Deepa Sharma et al. (247\u2013255) describe laboratory network expansion.Tanwi Trushna & Rajnarayan R Tiwari (281\u2013285) introduce a new research institute.Anne Marie Thow et al. (268\u2013275) examine safeguards in trade and investment agreements.Susan P Sparkes et al. (276\u2013280) show how political economy analysis can improve health programmes. Sophie Budge et al. (286\u2013288) track the evolution of environmental sanitation.Siem Zethof et al. (289\u2013291) use data on stillbirths to interpret caesarean section rates."} +{"text": "In the editorial section, Delia Enria et al. (610) call for research into the effects of each public health and social measure used by governments in the COVID-19 response. Saverio Bellizzi et al. (611) describe a national SARS-CoV-2 vaccination programme that includes migrants and refugees.In the news section, Tatum Anderson (614\u2013615) reports on the need for investment in mental health services in the context of the COVID-19 pandemic. Kazuto Kato talks to Gary Humphreys (616\u2013617) about the ethical and societal challenges posed by editing of the human genome. Katherine Kirkby et al. (627\u2013639) analyse subnational completion of diptheria-tetanus-pertussis courses.Kathryn Dovel et al. (618\u2013626) count visits and evaluate services.Nithima Sumpradit et al. (661\u2013673) describe implementation of the national strategic plan.Shanquan Chen & Rudolf N Cardinal (674\u2013679) describe the investments needed.Don E Willis & Pearl A McElfish (680\u2013681) note disproportionate morbidity among the Marshall Islands diaspora.Joseph Yaria et al. (640\u2013652) review the evidence for effectiveness of guidelines.Ignacio Neumann et al. (653\u2013660) argue for global access to direct oral anticoagulants.Lynn Lieberman Lawry et al. (682\u2013684) propose a framework for programme design."} +{"text": "Department of Statistics and Operations Research (DEIO), Universitat Polit\u00e8cnica de Catalunya BarcelonaTech (UPC), Barcelona, Spain\u2019 and new affiliation 5 added \u2018Institute of Mathematics of UPC - BarcelonaTech (IMTech), Barcelona, Spain\u2019 for Daniel Fernandez was missing. The original article has been corrected\u201d."} +{"text": "The original article (Muhl-Richardson et al.,"} +{"text": "Erratum zu:Bundesgesundheitsbl 202110.1007/s00103-021-03396-9In der urspr\u00fcnglichen Originalfassung des Artikels wurden einige Namen der Autorinnen und Autoren nicht korrekt wiedergegeben.Die Namen wurden nun korrigiert."} +{"text": "Voor u ligt weer het volgende nummer van Tijdschrift voor Urologie. Dit keer met de abstracts van de Najaarsvergadering van de NVU. We zijn er inmiddels aan gewend dat we elkaar minder vaak zien, dat de interactie nog steeds veelal digitaal is en dat er nog steeds wetenschap wordt bedreven. Interessant is het om te zien of de inhoud van de abstracts anders is geworden. De urologische praktijk w\u00e9l. Hoe? Dat weten we nog niet in detail. Andere prioriteiten, andere volgorde? Voor u ligt weer het volgende nummer van Tijdschrift voor Urologie. Dit keer met de abstracts van de Najaarsvergadering van de NVU. We zijn er inmiddels aan gewend dat we elkaar minder vaak zien, dat de interactie nog steeds veelal digitaal is en dat er nog steeds wetenschap wordt bedreven. Interessant is het om te zien of de inhoud van de abstracts anders is geworden. De urologische praktijk w\u00e9l. Hoe? Dat weten we nog niet in detail. Andere prioriteiten, andere volgorde?De arts-assistenten, arts-onderzoekers en urologen hebben weer zeer divers onderzoek opgestuurd. De wetenschappelijke commissie heeft zich weer van zijn taak gekweten en het meest relevante onderzoek geaccepteerd voor de vergadering, hoe hybride die dan ook wordt gehouden. Nog steeds is prostaatkanker ons nummer \u00e9\u00e9n interessegebied met zijn aparte sessie. Ook hierbinnen zijn er verschillende interessegebieden, maar die zijn nog wel vaak gerelateerd aan de PSMA/PET. Hopelijk zal er tijdens de jaarvergadering discussie ontstaan over hoe we die scan in de dagelijkse praktijk gaan implementeren.Weinig echter wordt COVID-19 binnen de urologie onderzocht. Geen enkele urologische aandoening is aan deze ziekte gerelateerd. Was hematurie bijvoorbeeld een gevolg van COVID-19, dan had het bestuur of de commissie externe betrekkingen regelmatig aan de vele talkshowtafels gezeten, en hadden we vast meer BN\u2019ers onder ons. Maar nu moeten we het doen met de onderlinge gesprekken tijdens de najaarsvergadering.Met dit nummer kunt u zich op die vergadering voorbereiden en de presentaties van kritische vragen en nuttige suggesties voorzien. Met elkaar kunnen we de abstracts laten uitgroeien tot een waar artikel, wellicht ook weer ter publicatie in dit tijdschrift. We zijn benieuwd, maar we zijn vooral benieuwd h\u00f3e we elkaar weer gaan zien. Terug naar het normaal? Laten we ervan uitgaan dat de ervaring van digitaal werken zich blijft vertalen naar meer mogelijkheden voor het vergaren van kennis die niet eindigt in een fileborrel. Wie weet.Nogmaals heel veel leesplezier en \u2018tot corona\u2019, tot computerscherm of fysiek in welke vorm dan ook.Peter F.A. Mulders \u2013 uroloog"} +{"text": "Multifunctional Nanomaterials: Synthesis, Properties and Applications\u201d, we published three review papers and nine original research articles.In this Special Issue \u201cIn the first article (review) , Si Amar2O3\u2013ZnO nanostructures towards NO2 gas sensing based on the control of appropriate material synergistic effects. Further, Zhikun Peng et al. [4, MnSO4 and FeSO4 on the partial hydrogenation of benzene over nano Ru-Based catalysts. The ninth published research article by Xiaoyu Yu et al. [In the fourth article (research), Rafael Bosch et al. reportedg et al. reportedu et al. , discussu et al. report aFurthermore, in the eleventh published research article, Yuan Tan et al. reported"} +{"text": "Een voorspeller van het aandeel mensen dat daadwerkelijk een vaccinatie zal nemen is de vaccinatiebereidheid onder de bevolking. Uit buitenlandse literatuur blijkt dat de vaccinatiebereidheid onder mensen met een lagere sociaaleconomische status lager ligt dan onder andere groepen. In deze bijdrage beschrijven we in hoeverre dit ook in Nederland het geval is en laten we zien hoe risicoperceptie, vertrouwen in de werking en veiligheid van het vaccin en gezondheidsvaardigheden hier mogelijk mee samenhangen. Tot slot belichten we een aantal interventiestrategie\u00ebn die positief aan de vaccinatiebereidheid onder laagopgeleiden kunnen bijdragen Sinds begin 2021 is vaccinatie een centraal onderdeel in de bestrijding van het COVID-19-virus. Om in te kunnen schatten hoeveel mensen zich willen laten vaccineren wordt gekeken naar de vaccinatiebereidheid. De vraag is hierbij in hoeverre deze bereidheid voor de verschillende subgroepen in de bevolking even hoog is. Buitenlandse onderzoeken laten zien dat mensen uit lagere sociaaleconomische groepen over het algemeen minder bereid zijn zich te laten vaccineren dan mensen uit hogere sociaaleconomische groepen. In onderzoeken in Groot-Brittanni\u00eb en de VS kwam bijvoorbeeld naar voren dat een laag inkomen samenhangt met negatieve opvattingen over het COVID-19-vaccin en dat de vaccinatiebereidheid toeneemt naarmate het opleidingsniveau stijgt [De WHO geeft gedragswetenschappen een belangrijke plaats in de bestrijding van pandemie\u00ebn, naast andere essenti\u00eble expertise, zoals virologie, epidemiologie en geneeskunde. In lijn daarmee werd in het voorjaar van 2020, tijdens de eerste golf van de COVID-19-pandemie, de Corona Gedragsunit opgericht, met als doel om\u00a0wetenschappelijke kennis en expertise over gedrag op geco\u00f6rdineerde wijze in te kunnen zetten bij de bestrijding van de coronapandemie . Naast lVoor deze bijdrage hebben we de tiende ronde van het vragenlijstonderzoek gebruikt. Tussen 10\u00a0en 14\u00a0februari namen 54.363\u00a0mensen in de leeftijd vanaf zestien jaar deel. Van hen was 36% man en 64% vrouw. In het vragenlijstonderzoek is een onderscheid gemaakt tussen laag- , middelbaar en hoogopgeleiden .Met de vraag \u2018Als u uitgenodigd wordt voor een vaccinatie tegen het coronavirus, wilt u zich dan laten vaccineren?\u2019 werd de vaccinatiebereidheid onder de deelnemers uitgevraagd. In totaal gaf 84% van de algehele bevolking aan bereid te zijn zich te laten vaccineren. Tabel\u00a0De vaccinatiebereidheid hangt binnen de opleidingsgroepen ook samen met leeftijd. Naarmate de leeftijd stijgt, wordt het verschil in vaccinatiebereidheid tussen opleidingsgroepen kleiner. Zo is in de leeftijdsgroep 25\u201339\u00a0jaar het verschil in vaccinatiebereidheid tussen laag- en hoogopgeleiden 23\u00a0procentpunt. Voor de groep 70-plussers is dit gedaald naar 5%.Uit gedragswetenschappelijk onderzoek weten we dat risicoperceptie een belangrijke factor is wat betreft de vaccinatiebereidheid . NaarmatNaast risicopercepties zijn ook vertrouwen in de werking van het vaccin en vertrouwen in de overheid belangrijke factoren die een rol spelen bij de vaccinatiebereidheid . Als er Om mensen in staat te stellen zelf een weloverwogen keuze te maken om zich wel of niet tegen COVID-19 te laten vaccineren, is het belangrijk dat mensen gebruik kunnen maken van betrouwbare en begrijpelijke informatie .Vaccin tegen Corona\u00a0\u2013 Pharos .Meer informatie over de ontwikkeling en inzet van interventies rond vaccinatiebereidheid is te vinden via de Corona Gedragsunit ("} +{"text": "Aquiles R Henriquez-Trujillo et al. (478) describe the impact of COVID-19 outbreaks among Amazonian indigenous people, in Ecuador. Mohammad I Khan et al. (479) make the case for increased vaccine manufacturing capacity in low- and middle-income countries.Gary Humphreys and Lynn Eaton (482\u2013483) report on adaptions made to clinical trials to ensure faster vaccine development for COVID-19. Renu Khanna talks to Andr\u00e9ia Azevedo Soares (484\u2013485) about improving policies for women\u2019s health.Aaron M Orkin et al. (514\u2013528) review the evidence for provision of emergency care by lay responders.Ting-Yu Yeh & Gregory P Contreras (486\u2013495) track viral evolution.Naseem Salahuddin et al. (506\u2013513) study a shorter post-exposure prophylaxis regimen.Maria Neufeld et al. (496\u2013505) adapt and validate WHO\u2019s questionnaire.Pier Luigi Sacco & Manlio De Domenico (529\u2013535) compare COVID-19 to previous shocks.Kumanan Wilson et al. (536\u2013538) argue for better role recognition and support.Eric Crosbie et al. (539\u2013540) review supply and demand strategies."} +{"text": "In 2008, toen de combinatiefuncties werden ingevoerd, kon niemand nog bevroeden dat deze functie, tegenwoordig de buurtsportcoach genoemd, een groot succes zou worden. Anno 2021 doen vrijwel alle gemeenten in Nederland mee met wat inmiddels de Brede Regeling Combinatiefuncties heet. Zo heeft ook Sint Eustatius een buurtsportcoach. De waarde van de buursportcoach voor het gaan en blijven bewegen van mensen heeft zich inmiddels bewezen. De Vereniging Sport en Gemeente (VSG), nauw gelieerd aan de Vereniging Nederlandse Gemeenten (VNG), speelt vanaf het begin een actieve en belangrijke co\u00f6rdinerende rol bij het implementeren van de functie buurtsportcoach in Nederland. Belangrijke succesfactor van de gehanteerde strategie is: werken vanuit vertrouwen, maatwerk en continu\u00efteit. In dit artikel wordt ingegaan op de verschillende regelingen rond de combinatiefuncties en de buurtsportcoach, de gehanteerde strategie om te komen waar we nu staan en perspectieven voor de nabije toekomst. Met de Impuls Brede scholen, Sport en Cultuur werden vier belangrijke doelstellingen nagestreefd [In het eerste jaar werden gemeenten vanuit de Impuls Brede scholen, Sport en Cultuur volledig gefinancierd door het rijk voor het aanstellen van combinatiefunctionarissen. Deze combinatiefunctionarissen hadden als taak om sport of cultuur te combineren met ten minste \u00e9\u00e9n andere sector, zoals onderwijs, zorg, welzijn of het bedrijfsleven. Vanaf het tweede jaar moesten gemeenten het grootste deel (circa\u00a060\u202f%) zelf meebetalen. Voor de eerste fase, vanaf 2007, was het doel dat in 2012 2.500 combinatiefunctionarissen werkzaam waren in de sectoren onderwijs, sport en cultuur [Vereniging Sport en Gemeenten (VSG) werd gevraagd de algemene en specifieke \u2013\u00a0op gemeenten gerichte\u00a0\u2013 ondersteuning van de Impuls Brede scholen, Sport en Cultuur uit te voeren. Van VSG werd verwacht om naast de sportsector, ook de cultuur- en onderwijssector op zowel landelijk als regionaal niveau te ondersteunen met onder andere kennisoverdracht en kennisuitwisseling. Daarnaast waren er specifieke ondersteuning en advisering per sector georganiseerd. Zo deed NOC*NSF de advisering en ondersteuning vanuit de sector sport, de VBS het onderwijs en de Cultuurformatie de cultuursector. VSG is sinds 2007 de algemeen projectleider en \u2018het gezicht\u2019 van de combinatiefuncties.Het huidige succes van de regeling is relatief eenvoudig tot stand gekomen. Vanaf het begin hebben de partijen samengewerkt vanuit onderling vertrouwen, hebben ze maatwerk geleverd en zijn ze steeds gericht geweest op continu\u00efteit van de regeling. Dit resulteerde in een gestage groei van het aantal combinatiefunctionarissen en later, buurtsportcoaches. In 2008 zijn de 31\u00a0grootste gemeenten van ons land gestart met combinatiefuncties voor het primair of voortgezet onderwijs. Het grootste gedeelte richtte zich op de sportsector en een kleiner aantal op de cultuursector. Elk volgend jaar deden meer gemeenten mee. In de loop van de tijd zijn de landelijke regelingen veranderd, net als de focus. In 2021 beschikken vrijwel alle gemeenten over buurtsportcoaches als een richtlijn te hanteren. Dit betekent dat gemeenten naar eigen inzicht meer of minder fte kunnen inzetten dan het toegewezen aantal, en daarmee rekening kunnen houden met het gewenste opleidingsniveau van de combinatiefunctionaris. Tot slot heeft de BRC een bredere focus dan voorgaande regelingen, wat gemeenten de ruimte biedt om de functionarissen in te zetten zoals ze dat noodzakelijk achten. In 2019 nemen 347 van de 355 gemeenten deel aan de BRC, wat neerkomt op 98\u00a0procent van alle Nederlandse gemeenten. In de volgende paragrafen blikken we terug op de factoren die bijgedragen hebben aan dit succes.Tot 2008, voordat de Impuls Brede scholen, Sport en Cultuur werd ingevoerd, waren er verschillende subsidies voor belangrijke projecten, zoals de Buurt Onderwijs en Sport-impuls, De Breedtesport Impuls, het Nationaal Actieplan Sport en Bewegen, en Meedoen Alle Jeugd door Sport. Kenmerkend voor deze projecten was dat gemeenten moesten voldoen aan de bijbehorende voorwaarden en criteria die waren opgesteld door de landelijke overheid, en daarop afgerekend werden. Wanneer gemeenten niet aan deze \u2018blauwdruk\u2019 voldeden, moesten ze het eerder verkregen subsidiegeld terugstorten. De blauwdruk had ook tot gevolg dat gemeenten in hun verantwoording naar het rijk niet konden opschrijven dat ze maatwerk hadden geboden. Het bieden van maatwerk betekent immers afwijken van de blauwdruk. In de praktijk werd natuurlijk wel maatwerk geboden. Dit betekende dus dat er geen inzicht was in het feitelijke resultaat en dat het geld mogelijk niet altijd op de bestemde plek terechtkwam. De ervaring leert dat het in de praktijk altijd even duurt voordat het geld van een regeling zo wordt benut als de bedoeling is. De genoemde projecten waren van te korte duur, waardoor projecten vaak al stopten voordat ze goed gingen lopen en er dus geen continu\u00efteit geboden kon worden.Medewerkers van de VNG, VWS directie sport, NOC*NSF en VSG hebben toen ingezet om meer te gaan werken vanuit vertrouwen. Dat paste ook in het tijdsbeeld. Het vierde kabinet Balkenende trad in 2007 aan en gebruikte het begrip \u2018vertrouwen\u2019 vaak. Gemeenten, als eerste en dichtstbijzijnde overheidsloket, hadden meer en meer de wens om maatwerk te (mogen) leveren voor hun burgers. Deze wens vormde ook een belangrijke basis voor de latere decentralisaties.In het overleg tussen de driehoek van rijksoverheid, NOC*NSF en VNG/VSG vatte de opvatting post dat om \u00e9cht het verschil te kunnen maken niet een zoveelste subsidie voor een project verstrekt moet worden, maar dat de verbinding tussen sport, cultuur en onderwijs mogelijk moet worden gemaakt op basis van de lokale behoeften, met de gemeente als co\u00f6rdinator en aanjager. In dit overleg werd het belang van vertrouwen benadrukt, omdat dat past bij het leveren van maatwerk op lokaal niveau. Dit betekende onder andere dat er vanuit de regeling zo min mogelijk regels voorgeschreven werden, dat er geen rode kaarten werden uitgedeeld als dingen anders liepen dan gepland, en dat gemeenten ook geen \u2018strafkorting\u2019 kregen, in de zin dat ze achteraf verkregen gelden moesten terugbetalen. Ook werden de regelingen structureel ingevoerd in plaats van projectmatig. Het gevolg van deze aanpak was dat het lokale enthousiasme voor deze combinatieregelingen groeide en dat de combinatieregelingen in de loop van de tijd werden uitgebreid naar alle leeftijdscategorie\u00ebn.De structurele inzet van de combinatieregelingen, en daarmee het bieden van continu\u00efteit aan gemeenten, is achteraf gezien bijzonder, omdat Nederland in 2008 te maken kreeg met een economische recessie. Hoewel gemeenten het financieel vaak moeilijk hadden en de verschillende kabinetten als gevolg van de economische recessie fors moesten bezuinigen, bleef er voor de combinatiefuncties en later aanvullend voor de buurtsportcoaches geld bijkomen. Een belangrijke reden is dat verschillende onderzoeken steeds opnieuw hebben aangetoond dat de combinatiefunctionaris bijdraagt aan het bevorderen van de gezondheid. Het gaat bijvoorbeeld om het bevorderen van de participatie aan sport, de effecten van bewegen op schoolprestaties, de betekenis van sport en bewegen voor scholieren, complexe ketenvraagstukken in de jeugdzorg en het verkleinen van gezondheidsachterstanden.Een tweede punt dat bijdroeg aan de continu\u00efteit van de combinatiefunctieregeling is dat gemeenten na verloop van tijd wel de financiering moesten blijven organiseren, maar niet als enige de combinatiefunctionaris hoefden te cofinancieren. Dit betekent dat bijvoorbeeld een sportbedrijf of welzijnsorganisatie een buurtsportcoach kon inzetten omdat deze organisatie daar zelf voordeel van had. Toen daar bovenop de decentralisaties in de jeugdzorg en de Wmo plaatsvonden, ontdekte men in die beleidsvelden al snel de kansen die het inzetten van een buurtsportcoach biedt. De buurtsportcoach werd meer en meer een onderdeel van een ketenaanpak.In de toen veel gebruikte zorgpiramide kwam het woord \u2018sport\u2019 in de nulde lijn, het voorliggende veld, merkwaardig nagenoeg niet voor. Tegelijkertijd besefte men wel steeds meer dat de ontwikkeling van de zorgkosten door participatie van datzelfde voorliggende veld teruggedrongen zou kunnen worden. Hoewel de strijd om het geld tussen enerzijds zorg en anderzijds preventie nog altijd op de klassieker Ajax-Feijenoord lijkt, is er steeds meer wetenschappelijk bewijs gekomen voor de social return on investment van sport en bewegen. Elke ingezette euro in sport en bewegen levert aantoonbaar een positief resultaat op in deelgebieden als arbeidsparticipatie, welbevinden (met lagere zorgvraag) en gezondheid .De noodzaak ontstond om meer inzicht te geven in de maatschappelijke waarde van sporten en bewegen. Het Kenniscentrum Sport en Bewegen heeft aan Rebel en het Mulier Instituut gevraagd de omvang van de SROI van sport en bewegen in Nederland te berekenen en de kosten en opbrengsten zo veel mogelijk in euro\u2019s uit te drukken . Deze SRSinds 2020 is een SROI-dashboard beschikbaar waarmee de lokale inzet vergeleken kan worden met wat er landelijk gemiddeld wordt ingezet. Zowel de SROI, als bijvoorbeeld de beweegrichtlijn, de score op beweegvriendelijke omgeving en de gemeentelijke sportbegroting per inwoner wordt in het dashboard inzichtelijk gemaakt en er kan een scenario worden doorgerekend. Desgewenst kan een gemeente de uitkomsten van de doorrekening vergelijken met die van een groep vergelijkbare gemeenten. Met de resultaten kan de gemeente het eigen beleid op onderdelen aanpassen. De SROI is als instrument inzetbaar om van tevoren een voorspelling te doen over de uitkomst van de inzet van geld voor sport en bewegen. Voor het beleidsveld sport en bewegen vormt dit dashboard een stevige professionaliseringsslag.De VNG en VSG hebben samen met de vele partners bewust ingezet op een brede ontwikkeling en versterking van de functie van de buurtsportcoach. Sinds 2008 zijn vele honderden bijeenkomsten georganiseerd, kleinschalig, dichtbij, lokaal en landelijk. Deze bijeenkomsten waren zowel gericht op de buurtsportcoaches zelf, als op de samenwerking met partners in andere beleidsvelden, zoals onderwijs, cultuur en zorg. Dit leidde tot verdere professionalisering van de buurtsportcoachfunctie, intersectoraal werken en werken op tactisch en strategisch niveau, en een differentiatie in het takenpakket van de buurtsportcoaches. Daar hoort ook een verruiming bij van de mogelijkheden tot verschillende salarisniveaus. Voor ongeveer de helft van de buurtsportcoaches is het bedrag in het rekenmodel van \u20ac\u202f50.000 voldoende om de salariskosten te dekken. Voor de andere helft zijn de salariskosten gelijk aan het normbedrag of liggen de kosten daarboven. In de bestuurlijke afspraken van 2019 is ruimte gecre\u00eberd om hier flexibel mee om te gaan.Deze ontwikkelingen hebben bijgedragen aan een betere lokale beweeginfrastructuur, waarbij de buurtsportcoaches een cruciale, stimulerende en aanjagende rol spelen. Mede daardoor kunnen de lokale sportakkoorden een succes worden (349 gemeenten doen mee!). Die infrastructuur is goud waard. Het maakt het mogelijk om lokaal samen met allerlei partijen, dus niet alleen de sportsector, \u00e9chte ketenaanpak na te streven en ook mensen te bereiken die kwetsbaar zijn en via de normale sportkanalen nauwelijks bereikt worden. Vroegsignalering is een begrip dat in de lokale en regionale akkoorden dan ook veelvuldig voorkomt. COVID-19 heeft ervoor gezorgd dat er veel meer oog is voor de kwetsbare burger en tevens het inzicht dat voldoende beweging en participatie aan sport en beweegactiviteiten voor iedereen van groot belang zijn.De combinatiefunctieregeling heeft zichzelf in de loop van de jaren bewezen . Dat wilDe zorgkosten lopen volgens de Volksgezondheid Toekomst Verkenning 2018 op naar 175 miljard in 2040 [De invoering van de Gecombineerde Leefstijlinterventie, kortweg de GLI, in het basispakket van de zorgverzekering loopt moeizaam en kan beter. Er ligt een enorme kans om individuele of groepsgewijze begeleiding van gezonde leefstijl te verbinden met een buurtsportcoach-plus\u00a0\u2013 een buurtsportcoach die gespecialiseerd is op alle onderdelen van de GLI \u00e9n bewegen. In het ene geval uitvoerend, in het andere co\u00f6rdinerend. Naast lokaal maatwerk wordt daarmee ook individueel maatwerk mogelijk gemaakt. En dan hebben we het niet meer over Ajax-Feijenoord tussen de zorg en preventie. Voor de toekomst van de combinatiefunctieregeling zou dit een volgende logische stap zijn."} +{"text": "Dear Editor,Several reports have concluded that smoking might be partly associated with a decreased risk of coronavirus disease-2019 (COVID-19) infection, and that the \"nicotinic hypothesis\" can be applied to the preventive and therapeutic strategies for COVID-19 conductFarsalinos et al. 2020) conduct020 conduRegarding disease progression in hospitalized patients with COVID-19 infection, Farsalinos et al. (2020) conductAnother meta-analysis presented the risk of poor clinical outcomes in COVID-19 patients who were former smokers. Simons et al. (2021) conductRegarding the \u201cnicotine hypothesis\u201d, Usman et al. (2020) revieweThe author declares no conflict of interest."} +{"text": "In the editorial section, Benjamin Mason Meier et al. (178) argue for a closer link between evidence and global health laws affecting travel in the context of circulating variants of concern. Ren Minghui and Feng Zhao (179) point out that mental health could be better prioritized in humanitarian, peacebuilding and development programmes. Samira Choudhury et al. (180) advocate for substantive research on health impacts of chemical contaminants in food.In the news section, Andr\u00e9ia Azevedo Soares (183\u2013184) reports on rights-based health service provision for migrants in Europe. Tina Musuya talks to Tatum Anderson (185\u2013186) about her work in community-led reforms to prevent violence against women.Md Mehedi Hasan et al. (196\u2013204) assess progress towards global nutrition targets.Prashant Jarhyan et al. (216\u2013230) conduct a systematic review to establish prevalence estimates.Paul Dietze et al. (187\u2013195) report outcomes of a cohort study.Katherine Heath et al. (231\u2013236) produce new estimates to inform surveillance and service provision.Patrick S Lungu et al. (205\u2013215) track continuity of services during the COVID-19 pandemic."} +{"text": "Het meten van pati\u00ebntervaringen geeft belangrijke inzichten in de kwaliteit van de Nederlandse gezondheidszorg. Het huidige onderzoek toetst in hoeverre de ervaren kwaliteit van zorg door de jaren heen is veranderd en hoe deze samenhangt met veranderingen in zorg en gezondheid tijdens de coronapandemie.Pati\u00ebntervaringen zijn verzameld met tevredenheidsoordelen en de kwaliteitsindicator PREM Chronische Zorg, onder een representatieve steekproef van mensen met een chronische ziekte. Trendanalyses (2016\u20132020) zijn uitgevoerd en verschillen tussen subgroepen zijn getoetst met Mann-Whitney U\u2011toetsen.De kwaliteit van de zorg wordt over het algemeen positief ervaren, ook tijdens de coronapandemie in het najaar van 2020. In dat jaar zijn mensen het minst tevreden over de afstemming tussen zorgverleners en over de preventieve begeleiding van hun ziekte tevreden). Trendanalyses laten zien dat de tevredenheid over preventieve begeleiding is gedaald en dat de tevredenheid over gezamenlijke besluitvorming door de jaren heen schommelt. Mensen die gevolgen van de coronapandemie ervaren voor hun zorg of gezondheid beoordelen aspecten van de gezondheidszorg minder positief dan diegenen die geen gevolgen ervaren.Het is belangrijk om aandacht te hebben voor pati\u00ebntervaringen met zorgprocessen, waarbij extra nadruk zou moeten liggen op informatie over preventie, ondersteuning bij veranderingen in de gezondheid en de behandeling tijdens de coronapandemie, en goede afstemming tussen zorgverleners.De online versie van dit artikel (10.1007/s12508-022-00329-y) bevat aanvullend materiaal, toegankelijk voor daartoe geautoriseerde gebruikers. In de gezondheidszorg voor mensen met een chronische ziekte staat een integrale en persoonsgerichte aanpak steeds vaker centraal. Idealiter werken hierbij verschillende zorgverleners uit verschillende disciplines samen rond de zorgvraag van de pati\u00ebnt , 2. Zorgpatient reported experience measures (PREM\u2019s) gebruikt, vragenlijsten die ervaringen met specifieke aspecten van zorgprocessen inzichtelijk maken [De afgelopen jaren is veel aandacht geweest voor de ontwikkeling en invoering van het meten van pati\u00ebntervaringen als kwaliteitsindicatoren in de zorg. Rapportcijfers worden bijvoorbeeld ingezet om tevredenheidsoordelen van pati\u00ebnten in kaart te brengen over ontvangen gezondheidszorg of individuele zorgverleners. Daarnaast worden steeds vaker jk maken \u20136. De PRjk maken . Deze vrjk maken , 8.De coronapandemie en de bijbehorende maatregelen hebben geleid tot veel veranderingen in de gezondheidszorg. Mensen met een chronische ziekte kregen te maken met onder andere afgeschaalde of uitgestelde zorg in het ziekenhuis, en aangepaste zorg en ondersteuning via bijvoorbeeld digitaal contact met zorgverleners. Tijdens de eerste coronagolf had een op de tien pati\u00ebnten te maken met uitgestelde behandelingen en digitale afspraken . Een kleDe coronapandemie heeft mogelijk ook gezorgd voor veranderingen in de gezondheid van mensen. Recent onderzoek van het Sociaal en Cultureel Planbureau laat zien dat vooral kwetsbare groepen een verslechtering in gezondheid rapporteren tijdens de coronapandemie, zoals ouderen, mensen met onderliggend lijden en personen met een lage sociaaleconomische status . Ook menHoe beoordelen mensen met een chronische ziekte de zorg en ondersteuning verleend door verschillende zorgverleners, en de kwaliteit van de belangrijkste zorgprocessen waarmee zij te maken hebben?Is er in de tweede coronagolf een verandering in de ervaren kwaliteit van zorg en ondersteuning zichtbaar onder mensen met een chronische ziekte, vergeleken met de situatie v\u00f3\u00f3r de pandemie?Is er een verschil in pati\u00ebntenervaringen van mensen die a)\u00a0wel of geen veranderde zorg tijdens de tweede coronagolf hebben ervaren en b)\u00a0wel of geen gezondheidsverandering tijdens de tweede coronagolf hebben ervaren?Het huidige onderzoek maakt inzichtelijk hoe mensen met een chronische ziekte de kwaliteit van zorg ervaren tijdens de tweede coronagolf in Nederland, in het najaar van 2020, vergeleken met de situatie v\u00f3\u00f3r de coronapandemie. Ook wordt gekeken in hoeverre ervaren kwaliteit van zorg samenhangt met veranderde zorg en ondersteuning, en mogelijke gezondheidsveranderingen tijdens de tweede coronagolf. De volgende onderzoeksvragen worden beantwoord:Er is gebruikgemaakt van gegevens uit het Nationaal Panel Chronisch zieken en Gehandicapten (NPCG) van het Nivel. Het NPCG is een landelijk representatief panel van ongeveer 3.500 zelfstandig wonende Nederlanders van 15\u00a0jaar en ouder. Panelleden hebben een medisch gediagnosticeerde chronische somatische ziekte en/of een lichamelijke beperking. Ze worden geworven via aselecte steekproeven van huisartsenpraktijken en bevolkingsonderzoek [Voor het huidige onderzoek zijn vragenlijstgegevens geselecteerd over de thema\u2019s kwaliteit van zorg en veranderingen in zorg en gezondheid tijdens de coronapandemie. Alleen gegevens van respondenten met ten minste \u00e9\u00e9n chronische ziekte zijn ge\u00efncludeerd. Er is gebruikgemaakt van een longitudinaal onderzoeksontwerp met herhaalde najaarsmetingen gedurende de periode 2016\u20132020. Het longitudinale ontwerp is aangevuld met een cross-sectioneel onderzoeksontwerp met gegevens uit 2020, om te toetsen hoe de kwaliteit van zorg wordt beoordeeld door mensen die tijdens de coronapandemie wel of geen veranderde zorg of gezondheid ervaarden.Ervaren kwaliteit van zorg en ondersteuning zijn in kaart gebracht via tevredenheidsoordelen over acht zorgverleners: huisarts, medisch specialist, praktijkondersteuner huisartsenzorg (POH), wijkverpleegkundige, gespecialiseerd verpleegkundige, thuiszorg, fysiotherapeut en apotheker. Aan panelleden is gevraagd om aan de zorgverlener(s) met wie het afgelopen jaar contact is geweest een rapportcijfer tussen de 0\u00a0(heel erg slecht) en 10\u00a0(uitstekend) toe te kennen. Ook is gevraagd een rapportcijfer toe te kennen aan de totale zorg die de persoon heeft ontvangen. De gegevens zijn jaarlijks beschikbaar van 2016\u20132020.Daarnaast is gebruikgemaakt van de kwaliteitsindicator PREM Chronische Zorg . Deze vrIn 2020 is aan respondenten gevraagd of de coronacrisis gevolgen heeft (gehad) voor de behandeling of professionele ondersteuning van hun chronische ziekte (antwoordoptie \u2018ja/nee\u2019). Mensen die gevolgen rapporteerden, konden aangeven welke gevolgen voor hen van toepassing waren: 1)\u00a0de zorgverlener heeft een of meer behandelafspraken afgezegd; 2)\u00a0de zorgverlener heeft de behandeling uitgesteld; 3)\u00a0de behandeling heeft digitaal plaatsgevonden; 4)\u00a0de ondersteuning thuis is verminderd of stopgezet; en 5)\u00a0ik heb er zelf voor gekozen om de behandeling niet door te laten gaan.In 2020 is aan respondenten gevraagd of zij sinds het begin van de coronacrisis veranderingen in hun gezondheid hebben opgemerkt. De mogelijke antwoordcategorie\u00ebn waren 1)\u00a0nee, mijn gezondheid is niet veranderd, 2)\u00a0ja, mijn gezondheid is verslechterd en 3)\u00a0ja, mijn gezondheid is verbeterd.De volgende achtergrondkenmerken zijn verzameld: geslacht, leeftijd en opleidingsniveau en mate van lichamelijke beperking als gevolg van een chronische ziekte. Het type diagnose en het aantal chronische ziekten is via ICPC-codes bij huisartsen verzameld en gekoppeld aan de overige achtergrondkenmerken en de vragenlijstgegevens .p-waarde kleiner dan 0,01 is statistisch significant. Ontbrekende waarden zijn niet meegenomen in de analyses.De statistische analyse is uitgevoerd met MLWIN\u00a02.30 voor onderzoeksvraag\u00a01 en\u00a02, en StataSE\u00a015.0 voor onderzoeksvraag\u00a03 , 13. VooBinnen de vijf metingen (2016\u20132020) liep het aantal respondenten uiteen van 1.197 tot 1.512 personen. De respons was gemiddeld 77% . In het meetjaar 2020 was iets meer dan de helft van de respondenten vrouw . De gemiddelde leeftijd was 66\u00a0jaar, waarbij de leeftijdscategorie van 65\u00a0tot en met 74\u00a0jaar het meest vertegenwoordigd was. Ongeveer een vierde van de respondenten had een laag opleidingsniveau, 29,1% had een hoge opleiding afgerond. Veel voorkomende chronische ziekten waren hart- en vaatziekten , diabetes en astma of COPD . Drie\u00ebnvijftig procent van de respondenten had \u00e9\u00e9n chronische ziekte, 18,5% had er drie of meer. Het merendeel had geen of lichte lichamelijke beperkingen . Zie digitaal aanvullende content, tabel\u00a01 voor alle achtergrondkenmerken.In 2020 beoordeelden mensen met een chronische ziekte de totale zorg die zij ontvingen als goed, met een gemiddeld rapportcijfer van\u00a07,8. Het cijfer was constant gedurende de periode 2016\u20132020. De gespecialiseerd verpleegkundige ontving de hoogste beoordeling in 2020 . Het gemiddelde cijfer van de gespecialiseerd verpleegkundige is toegenomen over de jaren . De jaren daarna werd de thuiszorg steeds beter beoordeeld, waarbij een 8,2 werd toegekend in 2020 (lineaire trend). Ook de rapportcijfers voor de fysiotherapeut en de medisch specialist stegen in 2020 respectievelijk naar een 8,2 en 8,0 (lineaire trends). De POH werd beoordeeld met een 7,8\u00a0gemiddeld over de periode 2016\u20132020. Het beoordelingscijfer schommelt door de jaren heen . Er zijn geen veranderingen zichtbaar in de beoordeling van de huisarts en de wijkverpleegkundige. In 2020 ontvingen zij respectievelijk een 8,0 en 7,9 gemiddeld. Zie digitaal aanvullende content, tabel\u00a03 voor alle beoordelingscijfers per meetmoment.In 2020 gaf de grote meerderheid van mensen met een chronische ziekte aan tevreden te zijn met zowel de bejegening en de voorlichting , als de deskundigheid van de zorgverlener had in het najaar van 2020 te maken met een of meer veranderingen in de professionele zorg of ondersteuning. De meest gerapporteerde veranderingen waren behandelafspraken die digitaal plaatsvonden , afgezegde behandelafspraken en uitgestelde behandelafspraken . De ervaren kwaliteit van zorg verschilt tussen mensen die wel of geen veranderingen hebben ondervonden voor hun behandeling of ondersteuning . Zo gaven zij vaker het hoogst mogelijke cijfer in vergelijking met diegenen die tijdens de coronacrisis wel veranderingen hebben gemerkt . Ook rapporteerde de groep die veranderingen in de zorg heeft ervaren minder vaak dat er preventieve begeleiding plaatsvond , dan de groep die geen veranderde zorg of ondersteuning heeft ervaren . De twee groepen verschillen daarnaast in hun kwaliteitsoordeel over de afstemming tussen zorgverleners. Van de mensen die geen veranderingen hebben ervaren vond 61% de afstemming tussen hun zorgverleners goed, in tegenstelling tot 51,1% van de mensen die wel gevolgen hebben ervaren .Een vierde van de mensen met een chronische ziekte . Mensen die te maken kregen met uitgestelde of digitale behandelafspraken waren iets minder vaak tevreden over de afstemming tussen zorgverleners .In een subanalyse is gekeken of er verschil is in beoordelingscijfers en kwaliteitsoordelen tussen mensen die wel of niet te maken kregen met a)\u00a0afgezegde behandelafspraken, b)\u00a0uitgestelde behandelafspraken en c)\u00a0digitale behandelafspraken (data niet weergegeven). Diegenen die te maken kregen met afgezegde behandelafspraken beoordeelden de zorg met een lager cijfer en waren iets minder vaker tevreden over de preventieve begeleiding (n\u202f=\u2009967). Circa een op de zes rapporteerde een verslechtering in gezondheid en een kleine minderheid rapporteerde een verbetering . Gezondheidsveranderingen hangen samen met ervaren kwaliteit van zorg . De twee groepen beoordeelden daarnaast de mate van afstemming tussen zorgverleners verschillend. Van de mensen die een verslechtering in gezondheid heeft ervaren, is 44,0% tevreden met de afstemming tussen zorgverleners, in tegenstelling tot 60,5% van de mensen die geen verslechtering in gezondheid rapporteerde .De meerderheid van de mensen met een chronische ziekte ervaarde in het najaar van 2020 geen verandering in gezondheid (83,9%, Het huidige onderzoek onder een representatieve groep van mensen met een chronische ziekte in Nederland laat zien dat de kwaliteit van de zorg en ondersteuning over het algemeen positief wordt ervaren, ook tijdens de coronapandemie in het najaar van 2020. Mensen met een chronische ziekte beoordelen de totale zorg die zij ontvangen in 2020 als goed, met een gemiddeld rapportcijfer van 7,8. Ook de tevredenheidscijfers voor individuele zorgverlener zijn over het algemeen hoog. Voor een deel van de zorgverleners, zoals de huisarts en wijkverpleegkundige, zijn de beoordelingen in de periode 2016 tot 2020 niet veranderd. Voor een ander deel van de zorgverleners, zoals de gespecialiseerd verpleegkundige en de medisch specialist, zetten de stijgende trends in beoordelingscijfers die sinds 2016 zichtbaar zijn, door.Voor een subgroep van de mensen lijken de coronapandemie en de bijbehorende maatregelen in 2020 invloed te hebben op hun ervaren kwaliteit van zorg. Mensen die te maken hebben met uitgestelde of veranderde zorg en ondersteuning beoordelen de zorg minder positief, dan mensen die geen gevolgen ervaren. Ook de groep die tijdens het najaar van 2020 een verslechtering in de gezondheidstoestand ervaart, beoordeelt de zorg minder positief. Het is mogelijk dat mensen door veranderingen in de zorg en gezondheid tijdens de coronapandemie minder tevreden zijn met de kwaliteit van de zorg. Het is ook mogelijk dat degenen die de kwaliteit van de zorg lager beoordelen al v\u00f3\u00f3r de coronapandemie meer gebruikmaakten van zorg en ondersteuning, waardoor ze extra geraakt zijn door de pandemie en -maatregelen. In hoeverre veranderingen in zorg en ondersteuning, gezondheidsveranderingen en de ervaren kwaliteit van zorg op elkaar inspelen behoeft vervolgonderzoek. Desondanks is het belangrijk om aandacht te schenken aan de mogelijke impact van de coronapandemie op iemands gezondheid en behandeling. Twee van de vijf mensen met een chronische ziekte geven in eerder onderzoek aan dat uitgestelde of verschoven zorg gevolgen voor ze heeft . Ander oZorgprocessen worden over het algemeen goed beoordeeld door mensen met een chronische ziekte. Dit komt overeen met eerdere onderzoeksbevindingen met het huidige panel . InternaOngeveer een derde van de mensen met een chronische ziekte is niet tevreden over de preventieve begeleiding voor hun ziekte. De tevredenheidscijfers laten hierbij een duidelijk dalende trend zien. Ook hier zou de coronapandemie een mogelijke verklaring voor de cijfers kunnen vormen. Het belang van een gezonde voeding en voldoende beweging, die onderdeel zijn van de preventie, zijn sinds de start van de coronapandemie benadrukt . De resuMensen met een chronische ziekte zijn het minst tevreden over de afstemming tussen zorgverleners; 36% van hen is niet tevreden. Het percentage is nog groter in de groep mensen die ervaren dat de coronacrisis gevolgen heeft voor hun zorg en ondersteuning, en hun gezondheid. De tevredenheid over de afstemming tussen zorgverleners is in 2020 niet anders dan in eerdere jaren. Al sinds enige jaren blijkt uit zowel nationaal als internationaal onderzoek dat een grote groep pati\u00ebnten ontevreden is over de afstemming tussen zorgverleners en de toegang tot multidisciplinaire zorg . Een op Het huidige onderzoek kent sterke punten, zoals het longitudinale karakter met meerdere meetjaren. Hierdoor is het mogelijk om uitspraken te doen over de ontwikkeling in ervaren kwaliteit van zorg en konden we in kaart brengen of een bepaalde trend die v\u00f3\u00f3r de coronapandemie zichtbaar werd tijdens het najaar van 2020 al dan niet verder doorzette. Door gebruik te maken van een groot panel met aselecte wervingsmethoden en wegingen zijn de uitkomsten van het onderzoek daarnaast generaliseerbaar naar de groep mensen met een chronische ziekte in Nederland.Het onderzoek kent ook beperkingen. Ongeveer een derde van de PREM-items bestond uit ontbrekende waarden. In het laatste meetjaar waren dit relatief meer ontbrekende waarden op de domeinen bejegening, voorlichting en deskundigheid dan in eerdere jaren. Op de domeinen gezamenlijke besluitvorming, preventieve begeleiding en afstemming zien we door de jaren heen schommelende ontbrekende waarden, maar in het algemeen relatief meer ontbrekende waarden dan op de andere drie domeinen. Het is belangrijk om te bedenken dat sommige stellingen van dit meetinstrument wellicht niet begrijpelijk waren of niet van toepassing waren op de situatie van een deel van de respondenten, bijvoorbeeld omdat ze niet te maken hadden met verschillende zorgverleners die zorg moesten afstemmen. De uiteindelijke aantallen waren echter voldoende om uitspraken over deze kwaliteitsindicator te doen. Tevens kunnen veranderingen in ervaren kwaliteit van zorg niet met zekerheid toegeschreven worden aan de coronapandemie. Aanvullend onderzoek is nodig om de effecten van de pandemie op de ervaren kwaliteit van zorg te specificeren en in context te plaatsen, bijvoorbeeld door kwalitatief onderzoek of door monitoronderzoek om te volgen of en hoe de huidige trends zich in de toekomst voortzetten.Het onderzoek laat zien dat mensen met een chronische ziekte over het algemeen positief zijn over de kwaliteit van de zorg in Nederland, ook tijdens de coronapandemie. Over sommige kwaliteitsprocessen is men minder tevreden, zoals de afstemming tussen zorgverleners en de preventieve begeleiding bij de ziekte. Er is een afname zichtbaar in de tevredenheid over adviezen over preventie die mensen krijgen. Veel mensen met een chronische ziekte hebben tijdens de coronapandemie te maken gekregen met veranderingen in de gezondheidszorg. Het lijkt belangrijk om aandacht te schenken aan pati\u00ebntervaringen met zorgprocessen, waarbij extra nadruk zou moeten liggen op informatie over preventie, ondersteuning bij veranderingen in de gezondheid en behandeling tijdens de coronapandemie en goede afstemming tussen zorgverleners."} +{"text": "Correction to: Psicol Refl Cr\u00edt 34, 11 (2021)https://doi.org/10.1186/s41155-021-00177-wFollowing publication of the original article (da Silva et al., The incorrect author name is: Fabriane Frota da Rocha Morgado.The correct author name is: Fabiane Frota da Rocha Morgado.The author group has been updated above and the original article (da Silva et al.,"} +{"text": "We thank Millen et al (Ada HeaFirst, Millen et al state thSecond, Millen et al state thLastly, Millen et al suggest"} +{"text": "De coronacrisis en de gevolgen die deze op de gezondheid van de Nederlandse bevolking heeft gaan de normale regionale onderzoeksaanpak te boven. Daarom heeft het Netwerk GOR-COVID-19 \u2013\u00a0dat bestaat uit GGD GHOR Nederland (namens de GGD\u2019en), RIVM, Nivel en ARQ Nationaal Psychotrauma Centrum\u00a0\u2013 het initiatief genomen voor een landelijk onderzoeksprogramma om de impact van de coronapandemie op de mentale en fysieke gezondheid van de Nederlandse bevolking op lange termijn te monitoren: de integrale Gezondheidsmonitor COVID-19. In dit artikel beschrijven we de achtergrond en opzet van deze gezondheidsmonitor, die erop is gericht onderzoeksbevindingen toepasbaar te maken voor praktijk en beleid, zowel lokaal als nationaal. Bij de organisatie van crisisbeheersing bij rampen en andere crises wordt uitgegaan van lokale capaciteit en regie. Als het gaat om het volgen van de gezondheidsgevolgen en risico\u2019s over de tijd, ligt dit niet anders. \u2018Gezondheidsonderzoek bij rampen\u2019 of kortweg \u2018GOR\u2019 is ge\u00ebnt op het verkrijgen van goed inzicht in gezondheid(srisico\u2019s) en veranderingen die optreden in de grillige tijdlijn van een ramp. Deze informatie is onmisbaar voor beleidsmakers en professionals. De publieke gezondheidsorganisatie, inclusief GOR, is in Nederland regionaal georganiseerd. De huidige crisis vergt echter een bovenregionale aanpak, ook als het gaat om het onderzoeken van de gezondheidsgevolgen. Daarom heeft het Netwerk GOR-COVID-19 het initiatief genomen tot een landelijk monitoringsprogramma om de gezondheidsgevolgen van de coronacrisis op een geharmoniseerde wijze te volgen. Het Netwerk GOR-COVID-19 bestaat uit GGD GHOR Nederland (namens de GGD\u2019en), RIVM, Nivel en ARQ Nationaal Psychotrauma Centrum. De \u2018integrale Gezondheidsmonitor COVID-19\u2019 \u2013\u00a0GOR op lokaal, regionaal en landelijk niveau, waarbij verschillende typen databronnen bijeengebracht worden\u00a0\u2013 heeft een looptijd van vijf jaar (2021\u20132025). Het project wordt gefinancierd door het ministerie van VWS en krijgt subsidie van ZonMw. In deze bijdrage beschrijven we de doelstellingen, achtergrond en opzet van deze gezondheidsmonitor. De wereldwijde COVID-19-pandemie houdt sinds maart 2020 ook Nederland in zijn greep. De pandemie heeft een groot beroep gedaan op de zorgcapaciteit en resulteerde in een serie maatregelen gericht op beheersing van de uitbraak. De samenleving ging op slot: men bleef thuis, hield afstand en bijna alle openbare gelegenheden \u2013\u00a0inclusief scholen\u00a0\u2013 werden een tijd lang gesloten. Op het moment van schrijven (oktober 2021) heeft de pandemie in Nederland geleid tot bijna 1,8\u00a0miljoen bevestigde besmettingen, en ruim 18,000 geregistreerde dodelijke coronaslachtoffers [Naast ge\u00efnfecteerde personen zijn ook veel mensen indirect getroffen door de coronacrisis\u00a0\u2013 de coronacrisis raakt de samenleving als geheel. De directe en indirecte gevolgen van het virus en de maatregelen die genomen worden om verspreiding van het virus te voorkomen vormen een mogelijke bedreiging voor de gezondheid en het welzijn van de bevolking op zowel korte als lange termijn.Allerlei gezondheidseffecten van infectie(s) met het COVID-19 virus en maatregelen die zijn genomen zijn direct waarneembaar. In de eerste plaats geldt dit voor mensen die na infectie milde of ernstige klachten ontwikkelen, soms zo ernstig dat een ziekenhuisopname of zelfs overlijden volgt. Een ander deel van deze mensen houdt na besmetting langdurige lichamelijke klachten en vermoeidheid. Ook zijn er mensen die met serieuze\u00a0\u2013 niet aan COVID-19 gerelateerde\u00a0\u2013 klachten de weg naar zorg niet of later hebben gevonden, omdat de zorg zich een periode primair heeft gericht op COVID-19-pati\u00ebnten. Daarnaast zijn er mensen die kampen met mentale problemen veroorzaakt of bestendigd door de pandemie , 3. UiteOm de gezondheidseffecten van deze crisis op verschillende groepen in verschillende fasen zoveel mogelijk te beperken is informatie nodig voor beleid en praktijk. Een goede beeld- en oordeelsvorming over welke zorg nodig is en waar versterking van de zorgcapaciteit en -organisatie gewenst is kan hier richting aan geven . HiervooZorginhoudelijk doel: bijdragen aan het goed afstemmen van zorg en behandeling van betrokkenen;Beleidsmatig/organisatorisch doel: bijdragen aan het afstemmen van beleid en maatregelen om in te spelen op de zorg- en ondersteuningsbehoeften van betrokkenen;Maatschappelijk doel: het afgeven van een signaal dat de gevolgen van de ramp of crisis goed in kaart worden gebracht;Wetenschappelijk doel: het bijdragen aan de kennisbasis omtrent de gevolgen van rampen en crises voor volgende gebeurtenissen.Het wetenschappelijk doel is nooit doorslaggevend bij de beslissing tot het inzetten van GOR, maar is wel belangrijk voor kennisopbouw in Nederland.GOR kan op verschillende manieren bijdragen aan de gezondheid van getroffenen. In hoofdlijnen zijn vier verschillende doelen te onderscheiden, 9:ZorgiTot nu toe is de ervaring met GOR in Nederland voornamelijk beperkt tot relatief kleinschalige, gelokaliseerde rampen. Het ging vrijwel altijd om \u2018flitsrampen\u2019, plotselinge gebeurtenissen met een onmiddellijke nasleep. Zo is gezondheidsonderzoek gestart na de vuurwerkramp in Enschede (2000), de Nieuwjaarsbrand in Volendam (2001), de poldercrash (2009) en de ramp met vlucht MH17 (2014). Het enige voorbeeld van longitudinaal GOR bij een langslepende crisis in Nederland betreft de aardbevingsproblematiek in Groningen .Vanuit de Wpg zijn de gemeenten verantwoordelijk voor het uitvoeren van GOR, in de praktijk is deze taak belegd bij de regionale GGD. De GGD adviseert in de gebruikelijke gang van zaken het bevoegd gezag \u2013\u00a0eventueel met ondersteuning van de expertgroep Nazorg van het centrum Gezondheid en Milieu van het RIVM\u00a0\u2013 over het wel of niet uitvoeren van GOR. Na een beslissing door het bevoegd gezag om GOR uit te voeren heeft de GGD een co\u00f6rdinerende rol bij het opzetten en uitvoeren van het onderzoek. Zoals gezegd vergt de huidige crisis vanwege de ingrijpende en landelijke aard van de impact echter een bovenregionale aanpak.De hoofddoelstelling van de integrale Gezondheidsmonitor COVID-19 is het bieden van een goede informatiebasis wat betreft de fysieke en mentale gezondheidseffecten van de COVID-19-crisis, om lokale en regionale bestuurders te kunnen adviseren en ondersteunen bij beleidsvorming, en het aanreiken van handelingsperspectieven.het verwerven van inzicht in de directe effecten van het coronavirus (COVID-19) op de gezondheid;het verwerven van inzicht in de effecten van de maatregelen gericht op de beheersing van de pandemie op de gezondheid;het bevorderen van de doorgeleiding van kennis en inzichten naar landelijke, regionale en lokale beleidmakers en zorgverleners.Subdoelstellingen zijn:De gezondheidsmonitor draagt bij aan het verkrijgen en doorgeleiden naar praktijk en beleid van inzicht in de effecten van de coronacrisis op de gezondheidssituatie van de bevolking. Om dit te optimaliseren hebben multidisciplinaire werkgroepen de taak om telkens wanneer nieuwe resultaten beschikbaar deze te duiden. Hieraan nemen vertegenwoordigers deel van de overheid, betrokken uitvoerende zorginstanties, het sociaal domein en instituten die een brug slaan tussen wetenschap, beleid en uitvoering. In de werkgroepen wordt een afweging gemaakt van bruikbaarheid van resultaten voor beleid en worden deze vertaald in concrete handelingsperspectieven. Ook op GGD-regioniveau worden dergelijke werkgroepen georganiseerd, om de vertaalslag te maken naar handelingsperspectieven voor lokale beleidsmakers en organisaties.Steeds staan dezelfde gezondheidsmaten centraal. Het gaat om klassieke gezondheidsmaten in GOR, zoals stemmingsklachten en depressies, angsten, su\u00efcidaliteit, middelengebruik, sociale en relationele problemen. Daarnaast worden ook non-specifieke klachten, chronische gezondheidsklachten en veranderingen in zorggebruik (inclusief medicatie) meegenomen. De klassieke GOR-gezondheidsmaten worden verder uitgebreid met andere gezondheidsthema\u2019s binnen de publieke gezondheid, zoals eenzaamheid, bewegen en regie over het eigen leven. Uiteraard wordt voortgebouwd op een brede kennisbasis van risico- en beschermende factoren.In de gezondheidsmonitor wordt waar mogelijk voortgebouwd op de bestaande regionale en landelijke onderzoeksinfrastructuur vanuit GGD\u2019en , RIVM en Nivel. Deze reguliere structuur vormt \u2013\u00a0in opgeschaalde vorm\u00a0\u2013 de basis voor de gezondheidsmonitor en levert cijfers op lokaal, regionaal en nationaal niveau, en maakt vergelijken mogelijk tussen gebieden en over de tijd, ook met de periode voorafgaand aan de uitbraak van het coronavirus in Nederland. Figuur\u00a0Lifelines). Ook wordt samenwerking gezocht met partijen die relevante doelgroepen volgen, bijvoorbeeld organisaties die betrokken zijn bij de hulpverlening aan OGGZ-doelgroepen.Wanneer gewenst of noodzakelijk \u2013\u00a0zoals bij het opzetten van de Openbare geestelijke gezondheidszorg (OGGZ)-monitor\u00a0\u2013 wordt samengewerkt met andere afdelingen van het RIVM , kennisinstituten of andere onderzoeksgroepen voorkomen binnen de populatie.Langcyclische monitoring geeft inzicht in het verloop van klachten en zorggebruik over de tijd, en factoren die daarbij een rol spelen, met oog voor risicogroepen en regionale/lokale verschillen of andere opvallende zaken.Voor de gezondheidsmonitor is gekozen voor twee vormen van monitoring, die min of meer tegelijk ontwikkeld en uitgevoerd worden: kortcyclische en langcyclische monitoring. Deze combinatie biedt inzicht in patronen en ontwikkelingen in gezondheid over de tijd en in de factoren die daarbij van invloed zijn, maar biedt ook de mogelijkheid om op basis van nieuw verkregen inzicht (snel) te schakelen tussen beleidsniveaus en op te schalen. Bij beide typen monitoring worden nieuwe data verzameld via vragenlijsten en wordt gebruikgemaakt van routinematig verkregen zorgregistratiedata, afkomstig van huisartsenpraktijken . Beide leveren andersoortige informatie op:Kortcyclische monitoring levert ongeveer vier keer per jaar een beperkte verzameling van ge\u00efnterpreteerde data op. De hoge frequentie van verschijnen draagt zorg voor inzicht in de actuele situatie. De data komen uit de NZR. Daarnaast worden zowel onder jeugd en jongvolwassenen samen (tot en met 25\u00a0jaar), als onder volwassenen door een extern onderzoeksbureau in opdracht van het netwerk metingen uitgezet in bestaande of nieuw op te richten representatieve panels.Aan de basis van langcyclische monitoring liggen primaire data. Alle GGD\u2019en in Nederland zijn hierbij betrokken. Zij zetten vragenlijsten uit voor verschillende leeftijdsgroepen: jeugd (klas\u00a02 en\u00a04 van het voortgezet onderwijs), jongvolwassenen en volwassenen en ouderen binnen hun bestaande panelstructuur. Voor de groep mensen die tot de OGGZ-doelgroep behoren wordt een nieuwe monitor ontwikkeld. Daarnaast worden data uit de NZR gekoppeld met CBS-data. Jaarlijks komen resultaten beschikbaar.Langcyclische monitoring levert resultaten op gemeenteniveau (voor grotere gemeenten op wijk-/buurtniveau) ten behoeve van lokaal beleid. Waar nodig worden deze aangevuld met registraties vanuit RIVM of externe onderzoeksgroepen. De langcyclische monitoring biedt meer diepgang door analyses van trends en patronen, vergelijkbaarheid op alle verschillende beleidsniveaus en koppeling van meerdere gegevensbronnen.Naast de monitoring wordt binnen het project jaarlijks een inventarisatie gehouden van lopende en afgeronde onderzoeken in Nederland met betrekking tot de gezondheidsgevolgen van de coronapandemie, en van internationale onderzoeksresultaten. De inzichten die hieruit voortkomen kunnen worden ingezet om de lang- en kortcyclische monitoring te optimaliseren, bijvoorbeeld als het gaat om het identificeren van groepen voor wie het risico op negatieve gezondheidsuitkomsten groter is. Daarnaast biedt deze inventarisatie een overzicht van kennishiaten en daarmee input voor de onderzoeksagenda voor verdiepend onderzoek. Tijdens de looptijd van de integrale Gezondheidsmonitor COVID-19 wordt twee keer een open subsidieoproep geplaatst (in 2022 en 2024) voor verdiepende onderzoeken die gebruikmaken van de verzamelde data.Crisis is kans. Het is een gevleugelde uitspraak. In dit artikel hebben we toegelicht hoe COVID-19 een acute aanleiding vormde om een brede monitoringsaanpak in te richten, die het zowel regionaal als landelijk mogelijk maakt om te anticiperen op gezondheidsrisico\u2019s en ontwikkelingen daarin. Dat betekent dat de komende jaren op verschillende niveaus steeds monitoringsresultaten worden ingebracht in het gesprek tussen wetenschappers, professionals en beleidsmakers (de eerste rapportages gericht op de jeugd worden op dit moment \u2013\u00a0eind 2021\u00a0\u2013 opgesteld).COVID-19 vormde een aanleiding, maar het ligt voor de hand dat de extra instrumenten, metingen en netwerken die tijdens de uitvoering van deze brede gezondheidsmonitor ontstaan, niet alleen ten goede zullen komen aan de GOR-taak in de toekomst, maar ook aan de reguliere gezondheidsmonitoring. Ook deze crisis legt immers bestaande gezondheidsrisico\u2019s en kwetsbaarheden bloot die ook los van deze crisis aandacht verdienen van beleidsmakers en professionals."} +{"text": "This month\u2019s theme issue focuses on policy and health system responses to the COVID-19 pandemic. In the editorial section, Viroj Tangcharoensathien and Tedros Adhanom Ghebreyesus (90) explain why the timing of the pandemic\u2019s end will be influenced by political choices. Oscar J Mujica et al. (91) emphasise the need to address health inequity in pandemic preparedness and response plans. Gary Humphreys (94\u201395) reports on mobile clinics and their operations, as part of the pandemic response. Uche Amazigo talks to Andr\u00e9ia Azevedo Soares (96\u201397) about the impact of the pandemic on neglected tropical disease control programmes. Sharika Nuzhat et al. (98\u2013107) analyse health and nutritional status data. Sachin S Singh & Lal Bahadur Singh (108\u2013114) document training programmes.Janani Muralikrishnan et al. (135\u2013143) measure access to eye care.Siti Helmyati et al. (144\u2013154) monitor continuity of service delivery.Flavia Riccardo et al. (161\u2013167) track the effectiveness of rapid risk assessment.Marianne Calnan et al. (127\u2013134) study continuity provided by telephone contact centres.Talerngsak Kanjanabuch & Krit Pongpirul (155\u2013160) assess consequences for patients.Is health equity improving?Jazmyn T Moore et al. (171\u2013173) et al detail strategies to reach minority groups. Nikki Gurley et al. (168\u2013170) capture policy developments.Genevie Fernandes et al. (174\u2013176) describe the requirements of resilient supply chains."} +{"text": "In the editorial section, Oladayo A Afolabi et al. (542) point to requirements for increased palliative care capacity in the African Region. Lena Morgon Banks et al. (543) describe the insurance coverage needed to improve access to health care for people with disabilities. In the news section, Gary Humphreys (546\u2013547) reports on the development of COVID-19 passes and the technical, ethical and social issues affecting their use. Amal Saif Al-Maani talks to Andr\u00e9ia Azevedo Soares (548\u2013549) about antimicrobial resistance and the need for multisectoral approaches to mitigating its development.Sabiha Essack (606\u2013608) notes the countries that cover both sectors in national action plans.\u00adBryony Simmons et al. (550\u2013561) measure progress towards antibiotic-use targets.Sonam Vijay et al. (562\u2013571) describe an integrated surveillance network.Suzanne M Connolly et al. (572\u2013582) review the evidence for lay counsellors.Hans J Overgaard et al. (583\u2013592) make the case for integrated disease management.Tracy Nau et al. (593\u2013602) propose a framework for legal strategies. Harman S Sandhu et al. (603\u2013605) consider the impact of animal husbandry on emerging pathogens."} +{"text": "In the editorial section, Alarcos Cieza et al. (686) encourage submissions to a theme issue on health policy and systems research for rehabilitation. Andrey Shukshin (689\u2013690) reports that tobacco control legislation is helping curb tobacco use in the Russian Federation. Agnes Kalibata talks to Gary Humphreys (691\u201362) about the need for multisectoral food system reform.Michelle L Giles et al. (739\u2013746) provide a case study on alignment of national recommendations.Leonor Guariguata et al. (722\u2013729) explore ways to address declining levels of physical activity.Anke Heitkamp et al. (693\u2013707) review the evidence for near-miss events.Marie Reilly & Bhavna Chohan (708\u2013714) describe pooled testing options for SARS-CoV-2.Sabine Vogler et al. (715\u2013721) detail joint medicine and vaccine procurement arrangements. Amiya Bhatia et al. (730\u2013738) argue for better prevention of violence against children.Sara Stratton et al. (747\u2013749) propose a move beyond wealth status and contraceptive use.Lukoye Atwoli et al. (750\u2013752) call for action to limit global temperature increases, restore biodiversity and protect health."} +{"text": "Title should read as \u201cHomology modeling and molecular dynamics simulation study of beta carbonic anhydrase of Ascaris lumbricoides\u201d in Yadav and Khandelwal, (2019) Bioinformation. 2019 15:572. PMID: 31719767."} +{"text": "Following publication of the original article (Chander et al. The original article (Chander et al."} +{"text": "In this month\u2019s editorial section Adnan A Hyder et al. (406) enumerate the effects of inequitable COVID-19 vaccine distribution. Stefan Listl et al. (407) argue that oral diseases and conditions need to be accounted for in universal health coverage.In the news section, Andr\u00e9ia Azevedo Soares (410\u2013411) reports on efforts to measure and address psychological distress in health care workers. Gary Humphreys interviews Iman Nuwayhid (412\u2013413) on the rebuilding of public health infrastructure in Beirut, Lebanon.Eleanor Turner-Moss et al. (464\u2013472) review evidence for associations with obesity.Cara Ebert & Janina I Steinert (429\u2013438) present prevalence estimates and describe risk factors.Ravindra Rao et al. (422\u2013428) study the logistics of mobile supply.Neeta Kumar et al. (446\u2013454) examine the utility of data held by individuals.Joyce Wamoyi et al. (475\u2013476) summarize the difficulties faced by people in informal urban settlements.Yusra Ribhi Shawar & Jeremy Shiffman (414\u2013421) examine the barriers to prioritization of a research agenda.Joseph S Puthumana et al. (439\u2013445) describe risk factors for burn injuries in children.Elia Gabarron et al. (455\u2013463) review the extent of misleading information related to COVID-19.Jared M Alswang et al. (473\u2013474) argue for the maintenance of neglected tropical disease programmes."} +{"text": "In the editorial section, Helen Clark et al. (846) introduce a handbook on social participation for universal health coverage. Wilson M Were and Anshu Banerjee (847) describe the revisions of WHO recommendations on managing hypoglycaemia in children. Angela K Shen et al. (848) explain how to ensure optimal supplies of vaccines for COVID-19. Samba Sow talks to Andr\u00e9ia Azevedo Soares (853\u2013854) about the need for regional approaches to medical product development, including treatments for mild and moderate COVID-19. Soares (851\u2013852) also reports on WHO\u2019s role in meeting unprecedented demand for effective supply chains in emergency response situations.Anna Mia Ekstr\u00f6m et al. (910\u2013912) introduce recent efforts to expand capacity.Mirriam Tyebally Fang et al. (892\u2013900) explore an unregulated industry.Kenneth Juma et al. (855\u2013864) estimate catastrophic expenditure and costs of treatment. Beth A Tippett Barr et al. (874\u2013882) propose a new indicator for HIV testing programmes.Justina O Seyi-Olajide et al. (883\u2013891) pilot the national surgical, obstetric, anaesthesia and nursing plan.Erica Reeve et al. (865\u2013873) analyse policy-makers\u2019 perspectives.Janet V Diaz et al. (901\u2013903) list current research questions. Helena Hildenwall and Fatsani Ngwalangwa (904\u2013906) call for updated guidance.Paul Ashford et al. (907\u2013909) detail issues of ethics, traceability and biovigilance."} +{"text": "In the editorial section, Shelly Chadha et al. (242) introduce WHO\u2019s first world report on hearing. Viroj Tangcharoensathien et al. (243) call for submissions to a theme issue on effective policy responses during the COVID-19 pandemic.In the news section, Andr\u00e9ia Azevedo Soares (246\u2013247) reports on initiatives aiming to improve access to insulin. Yemurai Machirori talks to Gary Humphreys (248\u2013249) about advocating for attention to the needs of people with type 1 diabetes.Jin Un Kim et al. (280\u2013286) argue that migrant populations need better access to diagnosis and treatment.Lynn Riggs et al. (259\u2013270) quantify disease caused by substandard housing.Shazia Moosa et al. (250\u2013258) study programme implementation.Padraig Lyons et al. (271\u2013279) examine what worked during the 2014-2016 Ebola outbreak.Mukta Sharma et al. (304\u2013311) review progress towards elimination targets for sexually transmittable diseases.Jennifer Cohn et al. (287\u2013295) present the case for triple elimination of mother-to-child transmission.Natthaprang Nittayasoot et al. (312\u2013318) explore elements of the COVID-19 response.Tommy Hana et al. (296\u2013303) examine the coverage of transgender health.Christine \u00c5rdal et al. (319\u2013320) argue for supply chain transparency."} +{"text": "The editorial team is grateful to all the experts who dedicated their time to peer-review our articles over the past 12 months. We wish to thank them and all those whose names are not listed below but gave helpful advice and support. We apologise to any reviewers whose names we may have missed.Our reviewers in 2021 were:Cornelia Adlhoch; Marco Ajelli; Mustafa Al-Haboubi; Abdullah A Algaissi; Franz Allerberger; Erik Alm; Juan Ambrosioni; Emmi Andersson; Denise Antona; Veli-Jukka Anttila; Patricia Antunes; Ludwig Apers; Ozgur Araz; Chris Archibald; Jakob Armann; Wouter Arrazola de O\u00f1ate; Mardjan Arvand; Tommi Asikainen; Agoritsa Baka; Tamas Bakonyi; Cyril Barbezange; Lucia Barcellini; Ian Barr; Ulrike Baum; Julien Beaut\u00e9; Senad Begi\u0107; Jaber Belkhiria; Leila Bell; Edward Belongia; Alexandre Belot; Cheryl Bennett; Fabian Berger; Zeno Bisoffi; Kaatje Bollaerts; Shelly Bolotin; Isabelle Bonmarin; Idesbald Boone; Timothy Booth; Robert Booy; Maria Jos\u00e9 Borrego; Michael Brandl; Arne Brants\u00e6ter; Jan Brauner; David Brett-Major; Anne Brisabois; Tom Britton; Norbert H. Brockmeyer; Kim Brolin; Jesca Brouwer; Alison Brown; Colin Brown; Udo Buchholz; Silke Buda; Niccol\u00f2 Buetti; Karina Butler; Zahid Butt; Merle Margarete B\u00f6hmer; Saverio Caini; S\u00e9bastien Calvignac-Spencer; Yehuda Carmeli; Eugenia Carrillo; Anne Carroll; Carlos Carvalho; Kelsie Cassell; Jesus Castilla Catalan; Simon Cauchemez; Dominique Caugant; Marco Cavaleri; Nathalie Charlotte; Remi Charrel; Tianmu Chen; Edward Choi; Eun Hwa Choi; Gerardo Chowell; Konstantin Chumakov; Annarita Ciccaglione; Heike Claus; Noelle Cocoros; Michelle Cole; Vittoria Colizza; Alex Cook; Geoffrey Coombs; Victor Corman; Laura Cornelissen; Ana Maria Azevedo Correia; Suzanne Cotter; Benjamin Cowling; Keith Crandall; Natasha Crowcroft; Fortunato D'Ancona; Gavin Dabrera; Andrei Dadu; Stephanie Dancer; Nicholas Davies; Brechje de Gier; Gaston De Serres; Gerard de Vries; Walter Demczuk; Aleksander Deptula; Phillippe Dewals; Vijaykrishna Dhanasekaran; Laura Di Domenico; Yaakov Dickstein; Joanne Dillon; Jac Dinnes; Lisa Domegan; G\u00e9 Donker; Yvonne Doyle; Aparna Dressler; Jan Drexler; Timothee Dub; Sebastian Duchene; Erwin Duizer; Dominic Dwyer; Robert Dyrdak; Michael Edelstein; Dirk Eggink; Hanne-Dorthe Emborg; Theresa Enkirch; Mirko Faber; Christopher Fairley; Luzhao Feng; Helen Fifer; Julie Figoni; Bastian Fischer; Brendan Flannery; Jannik Fonager; Arnaud Fontanet; Christina Frank; Ana Isabel Freitas; Luca Freschi; Ingrid Friesema; Shouichi Fujimoto; Emmanouil Galanakis; Irene Galani; Amiran Gamkrelidze; Karthik Gangavarapu; Patricia Garcia de Olalla; Patricia Garvey; Andrea Gervelmaeyer; Pere Godoy; Richard V. Goering; Edward Goldstein; Rok Grah; Devon Greyson; Roland Grunow; Thorolfur Gudnason; Claire Guinat; Rachel Gur-Arie; Jean-Paul Guthmann; Eric Haas; Walter Haas; Susan Hahne; Wanda Han; Pia Hardelid; Thomas Harder; David M. Hartley; Heli Harvala; Naoki Hasegawa; Joana Haussig; Qiushui He; Kristin Hegstad; Janneke Heijne; Wolfram Henn; Jean-Michel Heraud; Victoria Hernando; Thomas Hofmann; Yingfen Hsia; Jim F Huggett; Gwenda Hughes; Hilary Humphreys; Benedikt Huttner; Stefano Iacus; Derval Igoe; Michael G Ison; Michael Jackson; Stine Jakobsen; Sanjay Jayasinghe; David Jenkins; Cecilia Jernberg; Silvia Jimenez-Jorge; Kari Johansen; Tone Johansen; Jerker Jonsson; Fr\u00e9d\u00e9ric Jourdain; Irena Juki\u0107; Jaehun Jung; Annemarjut J\u00e4\u00e4skel\u00e4inen; Sarah Kabbani; Oliver Kacelnik; Kamran Kadkhoda; Bernard Kaic; Taishi Kayano; Winfried Kern; Ani Kevorkyan; Marian Killip; John Kinsman; Amir Kirolos; Esther Kissling; Jonas Klingstr\u00f6m; Marian Knight; Mirjam Knol; Anke Kohlenberg; Philipp Kohler; Axel Kola; Rolf Kramer; Annette Kraus; Gerard Krause; Manuel Krone; Andreas Kronenberg; Satu Kurkela; Jeff Kwong; Theresa Lamagni; Amparo Larrauri; Kevin B Laupland; Quentin Leclerc; Shui-Shan Lee; Clara Lehmann; Kathy Leung; Lanjuan Li; Pieter Libin; Andreas Lieberoth; Bruno Lina; Florence Lot; Yaniv Lustig; Michael F. Lynch; Frederik Lyngse; Theodore Lytras; Outi Lyytik\u00e4inen; Yiqun Ma; Gannon C.K. Mak; Michal Mandelboim; Davide Manissero; Elisa Mart\u00edn-Merino; Josefa Masa-Calles; Alberto Mateo Urdiales; Alberto Matteelli; Kathleen Mazor; Andrew McAuley; Andrea McCollum; Jim McMenamin; Eleanor Mcnamara; Michael McNeil; Gertjan Medema; Paula Meireles; Angeliki Melidou; Stefano Merler; Devon S Metcalf; Benjamin Meyer; Susana Monge; Dominique Monnet; Orna Mor; Roger Morbey; Alain Moren; Pedro l. Moro; Jo\u00ebl Mossong; Judith Mueller; Kristine M\u00f8rch; Marcel M\u00fcller; Sophie Nash; Eleni Nastouli; Pontus Naucler; Ndeindo Ndeikoundam Ngangro; Elsa Negro Calduch; Richard Neher; Nathalie Nicolay; Hiroshi Nishiura; Andreas Nitsche; Harold Noel; Robert Noland; Daan Notermans; Micha N\u00fcbling; Joan O'Donnell; Charlotte O'Halloran; Steve Oberste; Kazunori Oishi; Jason Ong; David Pace; W Paget; Nitika Pai; Scott Pallett; Takis Panagiotopoulos; Elisabetta Pandolfi; Junxiong Vincent Pang; Annalisa Pantosti; Dimitrios Paraskevis; Milosz Parczewski; Maria Pardos de la Gandara; Minal Patel; Samir Patel; Maria Paulke-Korinek; Pasi Penttinen; Andr\u00e9 Peralta-Santos; Javier Perez-Saez; John Pettersson; Naomi Petty-Saphon; Tung Phan; leanne pillay; Adriana Pistol; Denis Pi\u00e9rard; Diamantis Plachouras; Laurent Poirel; Jose A Ruiz Postigo; Isabelle Poujol; Wolfgang Preiser; \u202aGiovambattista Presti; Katarina Prosenc; Joan Puig-Barbera; Annalisa Quattrocchi; Sophie Quoilin; Public Rajatanavin; Venerando Rapisarda; Giovanni Ravasi; Carrie Reed; Ralf Reintjes; Ute Rexroth; Samuel Rhedin; Maria Luisa Ricci; Maximilian Riess; Michael Rigby; Thomas V. Riley; Julien Riou; Lorenz Risch; Annapaola Rizzoli; Emmanuel Robesyn; Marc Rondy; Werner Ruppitsch; Timothy Russell; Rachel Sacks-Davis; Katilyn Sadtler; Donald Salami; Paula Salmeron; Jet Sanders; Roy Sanderson; Frank Sandmann; Lawrence Sawchuk; Gianpaolo Scalia-Tomba; Samuel Scarpino; Astrid Blicher Schelde; Emily Scher; David Schwartz; Tatjana Schwarz; Matthew Scotch; Melissa Sharp; Lester Shulman; Reina Sikkema; Ryan Simpson; Vitali Sintchenko; Mary P. E. Slack; Morgane Solis; Gianfranco Spiteri; Megan Steain; Chen Stein-Zamir; Silvia Stringhini; Heino St\u00f6ver; Lorenzo Subissi; Carl Suetens; Jonathan Suk; Barbara Suligoi; Patrick Sullivan; Sheena Sullivan; P\u00e5l Suren; Pham Thi Mui; Daniel Thirion; Bruce Thorley; Karen Tiley; Jean-Fran\u00e7ois Timsit; Kelvin To; Maria Elena Tosti; Alberto Tozzi; Svetla Tsolova; Masaru Usui; Sophie Valkenburg; Maarten van Cleeff; Kees. C. van den Wijngaard; Marianne van der Sande; Esther Van Kleef; Thomas Vanwolleghem; Olli Vapalahti; Maria Louise Veimer Jensen; Lasse Vestergaard; Pauline Vetter; Jorge Vicente-Guijarro; Viktor von Wyl; Rengina Vorou; Jacco Wallinga; Michelle Waltenburg; Qiang Wang; Daniel Weinberger; Christian Wejse; Matthijs Welkers; Chard Richard Wells; Heather Whitaker; Andreas Widmer; Ursula Wiedermann; Annelies Wilder-Smith; Eloise Williams; Thomas Williams; Emma Wiltshire; Brita Askeland Winje; Carl-Heinz Wirsing von K\u00f6nig; Shell Wong; Takuya Yamagishi; Hsin-Chou Yang; Shangxin Yang; Qing Ye; Dominik Zenner; Walter Zingg; Phillip Zucs."} +{"text": "De meest voorkomende diabetesgerelateerde comorbiditeit is diabetische nefropathie of een andere vorm van nierschade.5,9 In 2020 zijn de voedingsrichtlijnen voor diabetespati\u00ebnten met nierschade herzien. Dit maakt dat de eerstelijns di\u00ebtist meer verantwoordelijkheid heeft gekregen voor deze pati\u00ebnten. Niet alleen is uit onderzoek gebleken dat het voor deze groep erg moeilijk is om een gepersonaliseerd dieet samen te stellen, van een di\u00ebtist wordt nu verwacht dat ze zelfstandiger stappen kan ondernemen. De richtlijnen onderstrepen dat samenwerking tussen di\u00ebtist en arts belangrijker is geworden. Hierbij is het doel om de diabetespati\u00ebnt met nierschade een betere kwaliteit van leven te geven.Het overgrote deel van de type 2-diabetespati\u00ebnten in Nederland wordt behandeld in de eerste lijn, waarbij de di\u00ebtist een prominente rol speelt. 47% van de di\u00ebtisten geeft aan dat diabetespati\u00ebnten hun grootste pati\u00ebntengroep vormen, blijkt uit onderzoek van de Nederlandse Vereniging van Di\u00ebtisten (NVD).5,6,7 Diabetische nefropathie wordt veroorzaakt door langdurige hyperglykemie. Het is een vorm van diabetesspecifieke nierschade die leidt tot toenemend eiwitverlies en uiteindelijk terminale nierinsuffici\u00ebntie. Kenmerkend voor deze aandoening is de hoge uitscheiding van albumine; 60% van het totale eiwitverlies in de urine is albumine. Bij andere nierziekten is dit circa 25%. Diabetische nefropathie gaat bijna altijd gepaard met hypertensie. In sommige gevallen ontwikkelt de diabetespati\u00ebnt eerst hypertensie, die later nierschade of diabetische nefropathie veroorzaakt. Het sterfterisico bij deze pati\u00ebnten neemt aanzienlijk toe als gevolg van hypertensie of andere vormen van hart- en vaatziekten.2,3,6Bij circa 12% van de type 2-diabetespati\u00ebnten wordt milde tot matige nierschade aangetoond op het moment van diagnose.5,6,8 Diabetespati\u00ebnten met ernstige nierschade of nierfalen hebben een verhoogd risico op hypoglykemie\u00ebn doordat bloedglucoseverlagende middelen minder goed worden uitgescheiden door de nieren.8 Dit resulteert vaak in het verlagen van de medicatie of zelfs het tijdelijk stoppen van de medicatie. Om te voorkomen dat een pati\u00ebnt verder achteruitgaat, wordt vanaf dat moment de nadruk nog meer op specifieke voedingsinterventies gelegd, zoals het beperken van eiwitten, zout en kalium. Hierdoor kan bij de pati\u00ebnt het gevoel ontstaan 'niks meer te mogen eten'; minder koolhydraten vanwege de glucose, minder eiwitten vanwege de nieren, minder groente en fruit vanwege het hoge kaliumgehalte.In eerste instantie zal er sprake zijn van nierinsuffici\u00ebntie, waarbij de nieren onvoldoende capaciteit hebben om afvalstoffen te filteren en uit te scheiden. Helaas is de prognose van diabetische nefropathie slecht; uiteindelijk zal 80% na 10 jaar nierfalen ontwikkelen.Richtlijnen Goede Voeding, waarbij er verschillende voedingsrichtlijnen bruikbaar zijn.9 De belangrijkste punten in de meeste richtlijnen voor diabetespati\u00ebnten met diabetische nefropathie of nierschade zijn gericht op:de glucoseregulatie;het voorkomen van overgewicht;de consumptie van micronutri\u00ebnten ;de consumptie van macronutri\u00ebnten ;vochtbeperking;alcoholgebruik.De voedingsadviezen voor diabetespati\u00ebnten met nierschade zijn gebaseerd op de 8Bovenstaande punten zijn enerzijds gekoppeld aan bloedglucoseregulatie en anderzijds aan de preventie van verergering van nierschade.Kidney Disease: Improving Global Outcomes zijn gericht op pati\u00ebnten met nieraandoeningen, waarbij er ook richting gegeven wordt aan diabetespati\u00ebnten met nierschade. Samenwerking tussen di\u00ebtist en arts blijft hierbij belangrijk; een gezondere levensstijl kan door een di\u00ebtist gemonitord worden en farmacologisch management wordt door de arts uitgevoerd. Bovendien kunnen beide partijen de pati\u00ebnt ondersteunen bij preventief zelfmanagement.De richtlijnen van de Belangrijke KDIGO-uitgangspunten zijn een gepersonaliseerd dieet met een zoutbeperking van < 2 gram/dag. Daarbij hanteert deze richtlijn een eiwit-inname van 0,8 gram eiwit/kg lichaamsgewicht/dag wanneer er geen sprake is van dialyse. Dit wordt verhoogd naar 1 tot 1,2 gram eiwit/kg lichaamsgewicht/dag bij pati\u00ebnten die wel gedialyseerd worden. Tevens wordt deze groep pati\u00ebnten aangeraden om minimaal 150 minuten per week te sporten. In de praktijk blijkt het erg moeilijk om een diabetespati\u00ebnt met nierschade te behandelen met voedingsinterventies. Voor nierpati\u00ebnten gelden namelijk andere voedingsrichtlijnen dan voor diabetespati\u00ebnten, en deze richtlijnen zijn soms in tegenspraak met elkaar. Zo kan het zijn dat de voeding die een nierpati\u00ebnt voorgeschreven krijgt een schadelijk effect heeft op het behouden van de juiste bloedglucosewaarden en vice versa. Omdat de bloedglucoseregulatie belangrijker wordt geacht dan de nierschade, is het advies aan di\u00ebtisten zich te richten op de voeding die een bloedglucoseregulerende werking heeft. Het gevolg is dat veel di\u00ebtisten zich conform de richtlijnen dan toch richten op de bloedglucose en niet op het voorkomen of verminderen van nierschade, omdat het simpelweg bijna onmogelijk is om dat met voeding te doen.5,8,9 Daarbij komt dat het gebruik van extra medicatie tegen eventuele andere aandoeningen de voeding nog meer aan banden kan leggen. Zo zijn er bepaalde medicijnen tegen hypertensie die de concentraties kalium in het bloed verhogen. Ondanks het feit dat die medicatie hypertensie tegengaat, is dit ongunstig voor de nieren.Het wordt nog moeilijker wanneer de diabetespati\u00ebnt gedialyseerd moet worden. Nierdialyse heeft een directe impact op de bloedglucoseconcentraties doordat glucose en ook insuline worden uitgescheiden tijdens de dialyse. Als gevolg hiervan moet het dieet worden aangepast. Zo is het bij nierschade essentieel de inname van kalium te verminderen, maar dit is vrijwel niet verenigbaar met de richtlijnen voor diabetespati\u00ebnten. Als de richtlijnen voor groente en fruit, peulvruchten en noten worden opgevolgd, gaat de inname van kalium namelijk verder omhoog. Zo kan een overschot aan kalium uiteindelijk leiden tot een hartstilstand. Kalium blijkt dan ook voor diabetespati\u00ebnten met nierschade de beperkende factor te zijn bij het naleven van de voedingsrichtlijnen en vormt dus een grote uitdaging voor de di\u00ebtist.NDF-voedingsrichtlijn Diabetes (2020).5 De NDF geeft daarnaast praktisch advies over ziektespecifieke drinkvoeding die bij sommige diabetespati\u00ebnten een uitkomst kan bieden, zoals bij pati\u00ebnten met nierschade. De nieuwe adviezen van de NDF voor deze pati\u00ebntengroep zijn werkbaar, maar wel wordt er meer ge\u00ebist van de eerstelijns di\u00ebtist. Een di\u00ebtist moet niet alleen de juiste voedingsinterventie kunnen toepassen, maar moet ook meer kennis vergaren over de aandoening(en) en medicijnen, wetenschappelijke artikelen lezen, extra opleiding volgen en soms lastige beslissingen nemen. De verschillende voedingsuitdagingen vragen om een multidisciplinaire aanpak. E\u00e9n van de belangrijkste punten in de vernieuwde NDF-voedingsrichtlijn Diabetes is dan ook dat een multidisciplinaire aanpak het risico op achteruitgang en het ontstaan van comorbiditeiten bij diabetespati\u00ebnten kan voorkomen. De belangrijkste taak van de di\u00ebtist is om preventief te werken en daarbij te voorkomen dat de aandoening verslechtert. Indien een pati\u00ebnt pas wordt doorgestuurd wanneer er al sprake is van een (sterke) achteruitgang en/of comorbiditeiten zoals nefropathie, wordt het werk van de di\u00ebtist een ware uitdaging.Praktische handvatten om nierschade zo veel mogelijk te beperken en de glucoseregulatie te handhaven, worden gegeven in de vernieuwde De voedingsdilemma's nemen toe wanneer de nierschade bij de diabetespati\u00ebnt verergert en wanneer er sprake is van meerdere aandoeningen tegelijk. Omdat de prognose van deze pati\u00ebntengroep slecht is, wordt het ook steeds moeilijker om een volwaardig dieet te kunnen samenstellen. Naarmate de nierschade verergert bij een diabetespati\u00ebnt zal er ook rekening gehouden moeten worden met verminderde eetlust en veranderende smaak. Een goede voedingsinterventie is in dergelijke gevallen uiterst belangrijk, maar ook bijzonder moeilijk. Uiteindelijk moet de di\u00ebtist een voedingskeuze maken die misschien niet meer in overeenstemming is met de dieetrichtlijnen. 2,8 Uit onderzoek is gebleken dat intensieve bloedglucoseregulerende behandelingen in deze groep niet zinvol zijn; er kan vaak alleen nog gekeken worden naar wat haalbaar is en waar de prioriteiten moeten liggen.Een ander dilemma doet zich voor bij de groeiende groep ouderen met diabetes en diabetische nefropathie. Deze kwetsbare groep heeft een korte levensverwachting.10,11 Het risico op ondervoeding in deze pati\u00ebntengroep is dan ook relatief groot wanneer er bepaalde beperkingen gelden voor de ene aandoening die interfereren met de andere aandoening. Dit laat ook het voorbeeld van kaliumreductie zien.1,2Uit de meeste voedingsrichtlijnen komt het belang van een persoonsgerichte aanpak naar voren. Het lastige is dat er geen eenduidige definitie bestaat van een diabetespati\u00ebnt met nierschade. Zeker niet voor pati\u00ebnten met bijkomende aandoeningen. Uit buitenlandse bronnen blijkt dat voedingsrichtlijnen vaak tekortschieten bij diabetische nefropathie en zeker in combinatie met andere comorbiditeiten.https://www.abbottnutritionprofessionals.nl/patientenonderzoekE\u00e9n van de oplossingen is het geven van ziektespecifieke voeding, zoals een drinkvoeding. In een onderzoek met 30 pati\u00ebnten kregen de pati\u00ebnten allen een gepersonaliseerd dieet voorgeschreven, waarbij in sommige gevallen de di\u00ebtist drinkvoeding adviseerde. Uit de resultaten van de kwalitatieve analyse bleek dat bij een groot deel van de pati\u00ebnten die drinkvoeding kregen een verbetering dan wel een stabilisatie van bloedwaarden zichtbaar werd. Hoe ouder de pati\u00ebnt, des te moeilijker het bleek om de nierfunctie stabiel te houden. Met name bij pati\u00ebnten > 70 jaar bleek de nierfunctie minder goed te verbeteren dan bij jongere pati\u00ebnten na het starten van de voedingsinterventie. Hoewel niet alle pati\u00ebnten dezelfde meetmomenten hadden, lieten de eGFR-resultaten zien dat met name mannen > 70 jaar kwetsbaarder zijn dan vrouwen van dezelfde leeftijd. In de leeftijdscategorie 60-69 jaar bleken vrouwen moeilijker te stabiliseren. Zie voor de onderzoeksopzet en de resultaten: 6 Een cruciale rol is weggelegd voor de di\u00ebtist om dit moment in te schatten en kwantitatief en kwalitatief te onderbouwen. Wanneer drinkvoeding werd ingezet, lieten de resultaten van de onderzochte groep pati\u00ebnten kwantitatieve en kwalitatieve gezondheidsverbeteringen zien. Ge\u00efnterviewde pati\u00ebnten gaven over het algemeen aan zich beter te voelen wanneer ze drinkvoeding namen.De huidige NDF-richtlijn adviseert drinkvoeding pas in te zetten wanneer iemand niet meer in staat is om met normale voedingsmiddelen in zijn voedingsbehoefte te voorzien.NDF-voedingsrichtlijn Diabetes (2020) heeft invloed op de huidige werkwijze van de di\u00ebtist. De di\u00ebtist moet meer zelf doen, meer kennis hebben over een aandoening en er zijn meer uitdagingen bij pati\u00ebnten met comorbiditeiten. De vernieuwde richtlijnen leggen meer druk op de di\u00ebtist. Ziektespecifieke voeding, zoals drinkvoeding, kan een hulpmiddel zijn bij deze groep pati\u00ebnten. De huidige richtlijnen adviseren drinkvoeding pas in te zetten wanneer iemand niet meer in staat is om met normale voedingsmiddelen in zijn voedingsbehoefte te voorzien.5 Hierbij is een cruciale rol weggelegd voor de di\u00ebtist om dit moment kwantitatief en kwalitatief te onderbouwen en in te schatten. Als artsen tijdig de diabetespati\u00ebnten doorsturen \u2212 nog voordat deze nierschade krijgen \u2212 en goed samenwerken met de di\u00ebtisten, kan dit in het voordeel van de diabetespati\u00ebnten werken. Dit resulteert in meer kwaliteit van leven, een beter dieet en minder risico's op het krijgen van ernstige nierschade of andere aandoeningen.Een substantieel deel van de diabetespati\u00ebnten krijgt te maken met nierschade, waarbij voedingsinterventies een belangrijke rol spelen om de achteruitgang in nierfunctie te vertragen. Het lastige is dat de voedingsrichtlijnen voor diabetespati\u00ebnten met nierschade moeilijk te verenigen zijn gezien de beperkingen in meerdere voedingsgroepen. Het verlagen van eiwitten in de voeding verlaagt het risico op verdere achteruitgang van de nieren. Kalium uit groente en fruit verlagen de bloeddruk en hebben een vochtafdrijvend effect. Indien er sprake is van diabetes en nierschade kunnen bepaalde eiwitrijke voedingsbronnen, maar ook groente en fruit, niet meer gegeten of minder gegeten worden. Bovendien heeft een diabetespati\u00ebnt al een beperking in koolhydraatinname. Hierdoor kan de pati\u00ebnt het gevoel krijgen niets meer te mogen eten. Naarmate de nierschade verergert bij een diabetespati\u00ebnt, zal er ook rekening gehouden moeten worden met verminderde eetlust en veranderde smaak. Een goede voedingsinterventie is in dergelijke gevallen uiterst belangrijk, maar ook bijzonder moeilijk. De nieuwe *Met dank aan dr. Suhail Khan, di\u00ebtist/nutritionist. Gebruikte data zijn de in het ziekenhuis gemeten bloedwaarden van pati\u00ebnten uit zijn praktijk.https://www.nvdietist.nl.Nederlandse Vereniging van Dietisten. https://www.volksgezondheidenzorg.info/onderwerp/chronische-aandoeningen-en-multimorbiditeit).Nielen MM., Poos MJJ. Chronische ziekten en multimorbiditeit; Cijfers & Context. Nivel (https://www.nivel.nl/nl/publicatie/comorbiditeit-bij-diabetes-mellitus.Nielen M. Comorbiditeit bij diabetes mellitus. Nivel. 2019. Sern\u00e9 E, Mourits P, van Strien M. Nationale Diabetes Registratie: Voor een toekomstbestendige en persoonsgerichte diabeteszorg in Nederland. Ned Tijdschr voor Diabetol. 2020. doi:10.1007/s12467-020-0131-2https://diabetesfederatie.nl/ndf-toolkit-persoonsgerichte-diabeteszorg/persoonsgerichte-voedingszorg.NDF Voedingsrichtlijn diabetes. 2020. Federatie Medisch Specialisten. Chronische nierschade. 2018.Hemmelder MH, van Balen J, Scherpbier N, Schenk PW, Tuut MK, Gansevoort RT. Herziening richtlijnen 'Chronische nierschade.' Ned Tijdschr Geneeskd. 2018. DNN. Chronische nierschade, diabetes en voeding. Dietist nierziekten. 2018.Gezondheidsraad. Richtlijnen goede voeding 2015. Gezondheidsraad. 2015.https://healthpolicy-watch.news.WHO. Ensuring people centred diabetes care during the Covid-19 pandemic. https://www.euro.who.int/__data/assets/pdf_file/0011/444791/Diabetes-care-during-COVID-19-eng.pdfWatch Health Policy. Covid-19. 2020. Wexler DJ, Grant RW, Wittenberg E, et al. Correlates of health-related quality of life in type 2 diabetes. Diabetologia. 2006. doi:10.1007/s00125-006-0249-9"} +{"text": "Elena Altieri et al. (754) introduce this month\u2019s theme on the use of behavioural science in health. Tedros Adhanom Ghebreyesus (755) provides an overview of the ways in which behavioural science can be applied.Gary Humphreys (758\u2013759) reports on systemic approaches to safer roads. Joyce Wamoyi talks to Andr\u00e9ia Azevedo Soares (760\u2013761) about attributes of behaviour change initiatives in sub-Saharan Africa.Solomon Aragie et al. (762\u2013772) study ways to change hygiene behaviours. Rajiv N Rimal et al. (773\u2013782) trial interventions to increase iron and folic acid consumption.Catherine Decouttere et al. (783\u2013794) explore vaccine hesitancy.Gavin George et al. (840\u2013842) strive to improve programme effectiveness.Sara Flanagan et al. (795\u2013804) report trial outcomes for family planning uptake.Alejandra L\u00f3pez G\u00f3mez et al. (843\u2013844) describe the effect of emerging evidence on public policies.Thomas Gadsden et al. (805\u2013818) review the evidence for performance-based incentives.Robin Schimmelpfennig et al. (819\u2013827) promote behavioural change for health.Rajiv N Rimal & Maria Knight Lapinski (828\u2013833) describe a conceptual approach to the field.Maria A Carrasco et al. (834\u2013836) outline research opportunities.April Monroe et al. (837\u2013839) study the intersection of behaviour and control efforts."} +{"text": "Peer Review Week 2021, the Eurosurveillance editorial team would like to send out a warm thank you to all our Rapid communications reviewers (90 experts in 2021 so far). Rapid communications have been a special feature over the last 25 years, through which the journal has had an impact and served the public health and communicable disease communities with timely and authoritative evidence for rapid public health action. Our peer reviewers are essential in shaping the quality and usefulness of the published articles, especially on short notice and we acknowledge all those who have reviewed rapid communications for us in 2021, despite their high workload during the ongoing COVID-19 pandemic:On the occasion of Cornelia Adlhoch; Marco Ajelli; Franz Allerberger; Erik Alm; Tommi Asikainen; Agoritsa Baka; Ian Barr; Senad Begi\u0107; Kaatje Bollaerts; Robert Booy; Maria Jos\u00e9 Borrego; Udo Buchholz; Saverio Caini; Jesus Castilla Catalan; Marco Cavaleri; Michelle Cole; Vittoria Colizza; Victor Corman; Laura Cornelissen; Benjamin Cowling; Natasha Crowcroft; Fortunato D'Ancona; Nicholas Davies; Brechje de Gier; Walter Demczuk; Aparna Dressler; Robert Dyrdak; Michael Edelstein; Dirk Eggink; Brendan Flannery; Arnaud Fontanet; Luca Freschi; Karthik Gangavarapu; Edward Goldstein; Thorolfur Gudnason; Eric Haas; Walter Haas; Thomas Harder; Kristin Hegstad; Derval Igoe; Michael G Ison; Michael Jackson; Kari Johansen; Annemarjut J\u00e4\u00e4skel\u00e4inen; Taishi Kayano; Marian Killip; Satu Kurkela; Amparo Larrauri; Kathy Leung; Bruno Lina; Michal Mandelboim; Andrea McCollum; Eleanor Mcnamara; Michael McNeil; Angeliki Melidou; Stefano Merler; Benjamin Meyer; Jo\u00ebl Mossong; Judith Mueller; Marcel M\u00fcller; Micha N\u00fcbling; Takis Panagiotopoulos; Maria Pardos de la Gandara; Pasi Penttinen; John Pettersson; Diamantis Plachouras; Katarina Prosenc; Giovanni Ravasi; Maximilian Riess; Julien Riou; Emmanuel Robesyn; Werner Ruppitsch; Gianpaolo Scalia-Tomba; Emily Scher; Gianfranco Spiteri; Carl Suetens; Jonathan Suk; Sheena Sullivan; Pham Thi Mui; Marianne van der Sande; Esther Van Kleef; Pauline Vetter; Qiang Wang; Matthijs Welkers; Eloise Williams; Thomas Williams; Emma Wiltshire; Shangxin Yang; Dominik Zenner; Walter Zingg.We extend this acknowledgement to everyone who reviewed articles for us during the last 25 years and an excuse to those who reviewed a rapid communication this year and who we may have missed in this list."} +{"text": "In the editorial section, Alana Officer et al. (710) highlight the hidden costs of aged-based discrimination against people. Andrew D Haddow & John P Woodall (711) emphasise a distinction between Zika and Spondweni viruses. Dale Gavlak (714\u2013715) reports on local training for mental health professionals in the eastern Mediterranean region. Epidemiologist Rosa Moreira tells Andr\u00e9ia Azevedo Soares (716\u2013717) how Angola managed to rein in its worst yellow fever epidemic in 30 years. Abdullah H Baqui et al. (752\u2013758) use trial data to estimate neonatal mortality.S\u00e9verine Henrard & Francis Arickx (779\u2013781) describe how joint price negotiations can apply to drugs for rare diseases. Paulo Paes de Andrade et al. (766\u2013771) examine the risk assessment done for a new method of mosquito control. Lucy A Perrone et al. (743\u2013751) describe a quality-improvement training programme for laboratory technicians. Shilu Tong et al. (759\u2013765) describe the population health implications of climate change.Mira Johri et al. (718\u2013727) study the feasibility of providing additional child health services during measles vaccination campaigns. Pankaj Bhatnagar et al. (728\u2013734) estimate the proportion of children receiving vaccines. Nandini Sharma et al. (777\u2013778) propose a comprehensive approach to prevention, recognition, and treatment of this occupational disease.Andreas Jahn et al. (772\u2013776) recount the steps taken to scale-up antiretroviral therapy.Jane Robertson et al. (735\u2013742) compare essential medicine lists for cancer treatment.Steve Kanters et al. (782\u2013784) describe how network meta-analysis can be used in clinical guidelines."} +{"text": "In the editorial section, Dermot Maher & Giorgio Cometto (786) explain why research on community-based health workers is needed to achieve the sustainable development goals. Naoko Ishikawa et al. (787) report elimination of mother-to-child transmission of HIV and syphilis in Cuba and Thailand.In the news section, Vijay Shankar Balakrishnan (790\u2013791) reports on the difficulties transgender people can encounter when accessing health services and interviews Yelzhan Birtanov (792\u2013793) about efforts to modernize the provision of health care in Kazakhstan.Andre Louis Wattiaux et al. (826\u2013834) quantify vaccine-preventable infections in indigenous adults.Juliana Sousa Soares de Ara\u00fajo et al. (835\u2013840) establish baselines and trends for congenital Zika syndrome.Belinda J Gabbe et al. (806\u2013816) use patients\u2019 reports to calculate the long-term effects of injuries. Mathieu JP Poirier et al. (817\u2013825) test children for multiple infections.Claire Leppold et al. (859\u2013860) explore longer-term public health considerations.Mar\u00eda Clara Restrepo-M\u00e9ndez et al. (794\u2013805) analyze trends of immunization coverage in 86 low- and middle- income countries. Justin Lessler et al. (841\u2013849) review the evidence for serological milestones in Zika virus infection.Hugo L\u00f3pez-Gatell et al. (850\u2013855) discuss the implications of dengue vaccine policies.Ahmad Reza Hosseinpoor & Nicole Bergen (856\u2013858) make a case for using area-based units of analysis."} +{"text": "Margaret Chan, Erik Solheim and Petteri Taalas (2) explain how the World Health Organization, the United Nations Environment Programme and the World Meteorological Organization work together on the environmental causes of ill health.Ishani Pathmanathan et al (3) explain why screening and treatment for tuberculosis is an integral part of care for people living with HIV.Jan Dirk Herbermann and Fiona Fleck (6\u20137) report on efforts to prevent both targeted and inadvertent destruction of health-care capacity in conflict settings. Peter Salama tells Fiona Fleck (8\u20139) how the World Health Organization\u2019s new emergencies programme is designed to help countries respond to crises. Christopher R Mikton et al. (36\u201348) propose research priorities.Wendy Rhymer and Rick Speare (10\u201317) evaluate countries\u2019 response to WHO\u2019s travel recommendations.Maureen Miller and Emily Hagan (62\u201368) describe other aspects of surveillance needed for pandemic prevention.Preeti Patel et al. (79\u201381) discuss motives for \u2013 and prevention of \u2013 attacks on healthcare facilities and staff.Bejoy Nambiar et al. (76\u201378) explain why quality improvement is an essential part of health-care delivery.Junfang Xu et al. (18\u201326) estimate the cost of illness attributable to dementia. G Suzanne A Smit et al. (27\u201335) measure costs and effectiveness of tuberculosis screening programmes.Elizabeth T Rogawski et al. (49\u201361) measure episodes of antibiotic treatment in children. Adam T Craig et al. (69\u201375) analyse transmission of Zika virus from 2007 to February 2015."} +{"text": "In the editorial section, Mahmood F Bhutta (314) introduces a campaign to improve conditions for workers who manufacture surgical instruments, gloves and textiles. Grania Brigden et al. (315) announce a new funding mechanism for research and development of tuberculosis medicines.In the news section, Tatum Anderson (318\u2013319) reports on efforts to prevent deaths from prescription opioids in Canada and the United States of America. Fiona Fleck (320\u2013321) interviews Svetlana Popova about her research on the global costs and consequences of maternal alcohol consumption during pregnancy.Sheikh Mohammed Shariful Islam et al. (382\u2013384) explore ways to improve national drug policies.Manoj Mohanan et al. (343\u2013352) study service provision for the treatment of diarrhoea and pneumonia in children.Miriam Rabkin et al. (353\u2013361) evaluate nurses\u2019 provision of HIV services. Alain Rakotoarisoa et al. (375\u2013381) describe a sentinel surveillance system. Margarete C Kulik et al. (362\u2013367) examine health, environmental and economic impacts of tobacco farming. Cameron Taylor et al. (322\u2013332) track inequities in ownership of mosquito nets. Karsten Lunze et al. (333\u2013342) assess the quality of care provided to children presenting with fever.H\u00e9l\u00e8ne Delisle et al. (385\u2013388) argue that more nutritionists are needed.Johanna Hanefeld et al. (368\u2013374) list problems and solutions for low- and middle-income countries."} +{"text": "In editorials this month, Shelly Chadha et al. (146) highlight future requirements for a public health response to hearing loss. Viroj Tangcharoensathien et al. (147) announce an upcoming theme issue on the prevention and control of noncommunicable diseases. Carlos Correa (148) comments on the use of flexibilities in the Agreement on Trade-Related Aspects of Intellectual Property Rights. Sima Barmania reports on architectural features of a medical research centre in Australia and a hospital in Norway, both designed to maximize the health of people who use them (151\u2013152). Carol Brayne talks to Tatum Anderson about research on the prevention and epidemiology of dementia (153\u2013154).Lelwala Guruge Thushani Shanika et al. (155\u2013164) observe the difference made by ward-based clinical pharmacists.Talat Ghane et al. (165\u2013172) collate reports of lead toxicity.Anne Marie Thow et al. (201\u2013210) examine fiscal policy measures designed to prevent non-communicable diseases.Martin Hitziger et al. (211\u2013218) trace implementation through the policy-making cycle. Abosede Adeniran et al. (222\u2013224) present a partnership for maternal, newborn and child health.Sandeep P. Kishore et al. (219\u2013221) consider what\u2019s needed for effective cardiovascular disease control.Ann Jagger et al. (173\u2013184) survey national policies from 148 countries. Ellen F.M. 't Hoen et al. (185\u2013193) review countries\u2019 use of flexibilities intended to ensure access to medicines.Rumina Hasan et al. (194\u2013200) proposes integration of control programmes."} +{"text": "Scientific Reports6: Article number: 3027110.1038/srep30271; published online: 07282016; updated: 02222017This Article contains errors in the Acknowledgements section:\u201cAbbott Vascular Int. (2011\u20132014), Amgen (2012\u20132018), AstraZeneca (2014\u20132017), Bayer (2013\u20132018), Boehringer Ingelheim (2013\u20132016), Boston Scientific (2010\u20132012), The Bristol Myers Squibb and Pfizer Alliance (2014\u20132016), The Alliance Daiichi Sankyo Europe GmbH and Eli Lilly and Company (2014\u20132017), Gedeon Richter Plc. (2014\u20132017), Menarini Int. Op. (2010\u20132012), MSD-Merck & Co. (2011\u20132014), Novartis Pharma AG (2014\u20132017), ResMed (2014\u20132016), Sanofi (2010\u20132011), SERVIER (2012\u20132018)\u201d.should read:\u201cAbbott Vascular Int. (2011\u20132014), Amgen (2009\u20132018), AstraZeneca (2014\u20132017), Bayer (2009\u20132018), Boehringer Ingelheim (2009\u20132016), Boston Scientific (2009\u20132012), The Bristol Myers Squibb and Pfizer Alliance (2011\u20132016), The Alliance Daiichi Sankyo Europe GmbH and Eli Lilly and Company (2011\u20132017), Gedeon Richter Plc. (2014\u20132017), Menarini Int. Op. (2009\u20132012), MSD-Merck & Co. (2011\u20132014), Novartis Pharma AG (2014\u20132017), ResMed (2014\u20132016), Sanofi (2009\u20132011), SERVIER (2009\u20132018)\u201d."} +{"text": "This month\u2019s issue has a special focus on emerging infectious diseases and antimicrobial resistance to accompany this year\u2019s Prince Mahidol Award Conference in Thailand. In editorials, Tedros Adhanom Ghebreyesus and Patricia Espinosa (78) draw attention to an initiative to protect people in small island developing states from the health impacts of climate change. Etienne Langlois et al. (79) argue that qualitative evidence should be used to improve health guidelines. In the news section, Sophie Cousins (82\u201383) reports on efforts to control bovine tuberculosis and prevent human exposure to infected meat and dairy products. Marion Koopmans tells Fiona Fleck (84\u201385) why the world needs a new global network to respond to emerging infectious diseases.Marc Poncin et al. (86\u201393) study the mass use of oral vaccines.Gumphol Wongsuvan et al. (94\u2013100) survey farmers on the quantities of antibiotics used when raising poultry for human consumption.Angkana Sommanustweechai et al. (101\u2013109) trace imports, manufacturing, distribution and regulation.Amitabh B Suthar et al. (110\u2013121) review lessons learnt in implementing the International Health Regulations.Julia Fitzner et al. (122\u2013128) examine clinical case definitions for influenza-like illness and severe acute respiratory infection.Victoria Y Fan et al. (129\u2013134) quantify expected losses from future pandemics.Jude Nwokike et al. (135\u2013137) argue for more stringent quality assurance measures.Daniel L Schar et al. (138\u2013140) present an investment case for the one health approach.Viroj Tangcharoensathien et al. (141\u2013144) consider interventions for improving antibiotic use."} +{"text": "Since publication of their article , the aut\u201cThe image data show radioactivity distribution in normal brain and cerebral metastases (enclosed in blue circle) (top panel) and are separated into blood (middle upper panel), non-blood (middle lower panel) and corresponding contrast-enhanced MRI images (bottom panel).\u201d"} +{"text": "In the editorial section, Ritu Sadana et al. (2) argue for the inclusion of older people in universal health coverage policies. Ramesh Krishnamurthy and Jason Hatton (3) describe how satellite imaging and other space technology can provide data for public health purposes. Jan Dirk Herberman (6) reports on measures taken to protect people and the environment from mercury exposure in Ghana, Malaysia and Thailand. Tom Frieden explains to Fiona Fleck (8) how concentrating on a few public health interventions to reduce noncommunicable diseases can help countries save the most lives.Ryan K McBain et al. (10\u201317) apply an activity-based costing exercise.Anamika Pandey et al. (18\u201328) track trends in catastrophic health expenditure over two decades.Jennifer F Friedman et al. (59\u201365) identify barriers to treatment of schistosomiasis during pregnancy.Aimable Mbituyumuremyi et al. (51\u201358) propose a framework for the national response for Hepatitis C infections.Karel Blondeel et al. (29\u201341) review evidence of violent acts motivated by perceptions of sexual orientation and gender identity.Shyama Kuruvilla et al. (42\u201350) look for ways to use the sustainable development goals.Effy Vayena et al. (66\u201368) explore policy implications of big data for the health sector.Benjamin H Chi et al. (69\u201371) summarize attempts to engage both parents during the childbearing year."} +{"text": "This month\u2019s issue has a special focus on healthy ageing, as introduced by John R Beard et al. (730) in the first editorial. Dulce M Cruz-Oliver et al. (731) expand on this theme with a discussion of end-of-life care in low and middle-income countries. Vijay Shankar Balakrishnan and Fiona Fleck (734-735) report on efforts to improve training and support for caregivers of people with dementia in Ghana, India and Lebanon. Andr\u00e9ia Azevedo Soares (736\u2013737) interviews Yuji Kuroiwa about technological and political approaches to the ageing population in Japan. Ray Serrano et al. (788\u2013790) examine filial support laws for fairness and reciprocity.Hannah H Leslie et al. (738\u2013749) assess service readiness of health facilities.Peter Lloyd-Sherlock et al. (774\u2013778) consider volunteer provision of long-term care.Sigvard M\u00f6lstad et al. (764\u2013773) collate lessons learnt from two decades of preventing antimicrobial resistance. Riaz Aziz et al. (779\u2013783) report tetanus deaths in men. Belinda Bennett et al. (750\u2013755) explore the implications of technology used to assist elderly people.Islene Araujo de Carvalho et al. (756\u2013763) describe WHO\u2019s approach to transforming health services.Frank Pega et al. (784\u2013787) explain the need to monitor action on the social determinants of health.Sridhar Venkatapuram et al. (791\u2013792) question assumptions about chronological and functional ageing."} +{"text": "Parasite in 2015 and 2016 and thank them for their help. The rejection rate has been about 50% since 2013. The mean time from submission to final publication was 104 days in 2016. The Impact Factor of Parasite has risen from 0.822 (published in 2014) to 1.781 (published in 2016). All papers published in Parasite and its predecessor Annales de Parasitologie Humaine et Compar\u00e9e since 1985 are freely available from the website of the journal.The Editors provide a list of the 237 reviewers from 43 countries who assessed manuscripts for Parasite. The success of the journal depends upon their care and competence and their conscientiousness is much appreciated.The Editor-in-Chief would like to thank all the reviewers who have given so generously of their time to assess manuscripts submitted to 237 reviewers from 43 countries are listed below. Names are in alphabetical order. Some have reviewed more than one manuscript. Note that for 2013\u20132014, the list included 292 reviewers from 51 countries [For 2015 and 2016, our Parasite has been about 50% since January 2013 and this list includes reviewers who evaluated papers which were not published.The rejection rate of Parasite are final versions: we do not publish uncorrected manuscripts or preliminary versions. Once published in Parasite, papers are in PubMed generally within 24\u00a0h still continues to be the key metric used by the academic community as a measure of quality of a journal. We are happy to show that the IF of Parasite has greaParasite has included Altmetric scores for all papers, including the papers published before this date. We are very pleased to see that many papers published in Parasite have received significant Altmetric scores, and have impressive download counts. All this information is visible for each paper from our website.In addition to the Impact Factor, other metrics are also becoming important. Academics and researchers are now evaluating not only the Journal as a complete entity, but also individual article metrics. Since 2015, Parasite are published in English. While we acknowledge the international role of English in the scientific community, we also respect other cultures and communities. In addition to the English abstract, all papers have a French abstract and on various occasions we have also added an Arabic or a Chinese abstract, and we are open to suggestions for future papers.All papers in Parasite have been downloaded from virtually every country on the planet, with more than 240,000 downloaded from our website in 2016, plus many more from other platforms.While the proportion of French authors is stable, we have experienced a tremendous increase in submission of papers from China, from 2% of submitted papers in 2013 to 18% in 2016. Papers from Parasite is the successor to Annales de Parasitologie Humaine et Compar\u00e9e, which was published from 1923 to 1993. We have now scanned and uploaded all papers published since 1985, and we intend to do more this year. Of course, all these papers are free to download with no pay-wall: we do not charge our readers $40 for a paper published 30\u00a0years ago!Parasite are full gold open-access papers with free immediate access for all to read. They are also available from other platforms such as PubMed Central, Europe PubMed Central, DOAJ and Science Open. In addition to traditional texts and images, we can publish movies and other media. All papers are available in html, PDF and even ePub formats to be readable on mobile devices.All papers published in Rob Adlard \u00c8ve Afonso (France)Bulent Alten (Turkey)Cristian A. Alvarez Rojas Samer Angelone-Alasaad (Switzerland)Karim Aoun (Tunisia)Andrea Apolloni (France)Thomas Barth (Germany)Christian Bauer (Germany)Ian Beveridge Oleg Blagosklonov (France)Peter Boag Franck Bou\u00e9 (France)Nacim Bouheraoua (France)Denis Boulanger (France)Gilles Bourdoiseau (France)J\u00e9r\u00e9my Bouyer (France)Rodney A. Bray (United Kingdom)Reginhaldo Brazil (Brazil)Emanuele Brianti Alessandro Broglia Joanna Browne Magdal\u00e9na Bru\u0148ansk\u00e1 (Slovakia)Nophawan Bunchu (Thailand)Jacques Cabaret (France)Oscar Cabez\u00f3n (Spain)Ermanno Candolfi (France)Luis Cardoso Yannick Caron (Belgium)S\u00f3crates C.H. Cavalcanti (Brazil)Ivan Cepicka (Czech Republic)Gabriela Certad (France)Piotr Ceryngier (Poland)Amira Chaabane (Tunisia)Qijun Chen (China)Jia-Xu Chen (China)Jia Chen (China)Eirini Christaki (Greece)Theresa Coetzer (South Africa)Simone Cohen (Brazil)Douglas Colwell (Canada)Melissa Conrad (United States)Barbara Conti Micha\u0142 Czopowicz (Poland)In\u00e1cio Domingos da Silva Neto (Brazil)Marie-Laure Dard\u00e9 (France)Arwid Daugschies (Germany)Isaure de Buron (United States)Reginald De Deken (Belgium)Johan F. De Jonckheere (Belgium)Harry De Koning (United Kingdom)Laurence Delhaes (France)J\u00e9r\u00f4me Depaquit (France)Peter Deplazes (Switzerland)Angela Di Cesare Mar\u00eda Celina Digiani (Argentina)Bilal Dik (Turkey)Isabelle Dimier-Poisson (France)Brent Dixon (Canada)Olgica \u0110urkovi\u0107-\u0110akovi\u0107 (Serbia)Stephen Doggett Philippe Dorchies (France)Gilles Dreyfuss (France)Jitender Dubey (United States)Jean Dupouy-Camet (France)Olivier Duron (France)Itziar Estensoro (Spain)Bart Everts (Netherlands)Jorge Falc\u00f3n-Ordaz (Mexico)Alessandra Falleni Anna Falt\u00fdnkov\u00e1 (Czech Republic)Vinicios Ferreira-de-Freitas (United States)Hubert Fert\u00e9 (France)William Font (United States)Elena Forte Michel Franc (France)Jos\u00e9 Ramon Franco (Switzerland)Claire Fuller (United Kingdom)Terry Galloway (Canada)Andr\u00e9 Garcia (Benin)Scott Gardner (United States)Claudio Genchi Dirk Geysen (Belgium)Mohamed Gharbi (Tunisia)Lynda Gibbons David Gibson (United Kingdom)Pooria Gill (Iran)David Gonz\u00e1lez-Sol\u00eds (Mexico)Bruno Gottstein (Switzerland)Philippe Grellier (France)Fr\u00e9d\u00e9ric Grenouillet (France)Norman Grim (United States)Zemao Gu (China)Ankita Gupta (India)Nabil Haddad (Lebanon)Ali Halajian (South Africa)Sarah Hamer (United States)Gabriel Hamer (United States)Clare Hamilton (United Kingdom)Majid Fasihi Harandi (Iran)Epco Hasker (Belgium)Louise Heaton (United Kingdom)Martin Heyworth (United States)Herv\u00e9 Hoste (France)Aurore Hounto (Benin)Sandrine Houz\u00e9 (France)Shaomin Hu (United States)Sung-Tae Hung (Korea)Seth Irish (United States)Tadashi Itagaki (Japan)Armando Jardim (Canada)Minjun Ji (China)Quentin Jossart (Belgium)Jean-Lou Justine (France)Zakaria Kambris (Lebanon)Saumyakanti Khatua (India)Marta Kicia (Poland)Muza Kirju\u0161ina (Latvia)Jovin Kitau (Tanzania)Seiki Kobayashi (Japan)Ismail Soner Koltas (Turkey)Jan Kopeck\u00fd (Czech Republic)Alexei Kostygov (Czech Republic)Michail Kotsyfakis (Czech Republic)Michael Kristensen (Denmark)Delane Kritsky (United States)Katrin Kuhls (Germany)Karien Labuschagne (South Africa)Gordon Langsley (France)Claire Laugier (France)Antti Lavikainen (Finland)Claudio Lazzari (France)Sylvie Lecollinet (France)Xiangrui Li (China)Quan Liu (China)Xiao-gang Liu (China)Gabriella Lo Verde Philippe Loiseau (France)Jacob Lorenzo-Morales (Spain)Hannelore Lotter (Germany)Eugene Lyons (United States)Sutherland Maciver (United Kingdom)Helena Maltezou (Greece)Francesca Mancianti Tanguy Marcotty (Belgium)Davis Mark (United States)Justin Masumu (Congo)Chris McAllister (United States)Catherine McCann (United Kingdom)Emily McDonald (United States)Antonio Menezes da Silva Jean Menotti (France)Alessandro Minelli David Modr\u00fd (Czech Republic)Paul Monis Scott Monks (Mexico)Jos\u00e9 Alberto Montoya-Alonso (Spain)Serge Morand (France)Luciano Andrade Moreira (Brazil)Ralf Mueller (Germany)Beat M\u00fclhaupt (Switzerland)John-Paul Mutebi (United States)Changshen Ning (China)Matthew Nolan (United Kingdom)Francesco Nugnes Emiliano Ocampo (Argentina)Takahiro Ohnishi (Japan)Susumu Ohtsuka (Japan)Laor Orshan (Israel)Domenico Otranto Elias Papadopoulos (Greece)Carine Paraud (France)Philippe Parola (France)Steven Peck (United States)Herv\u00e9 Pelloux (France)Gerardo P\u00e9rez Ponce de Le\u00f3n (Mexico)Bernard Pesson (France)Kl\u00e1ra Petr\u017eelkov\u00e1 (Czech Republic)Kurt Pfister (Germany)Davies Pfukenyi (Zimbabwe)Larisa Poddubnaya (Russian Federation)Bruno Polack (France)Robert Poulin Edoardo Pozio Iva P\u0159ikrylov\u00e1 (Czech Republic)Heather Proctor (Canada)Nicolas Puillandre (France)St\u00e9phane Ranque (France)Eva \u0158ehulkov\u00e1 (Czech Republic)Klaus Reinhardt (Germany)Mickael Riou (France)Joao Rodrigues Brice Rotureau (France)Urmas Saarma (Estonia)Oscar Salomon (Argentina)Vittorio Sambri Rodrigo Sanabria (Argentina)Claudia Santos (Brazil)Roy Sawyer (United States)Tomas Scholz (Czech Republic)Roshanak Tolouei Semnani (United States)Jeffrey Shaw (Brazil)Sara Shayan (Iran)Myeong Heon Shin (Korea)Gustave Simo (Cameroon)Tina Skinner-Adams Nico Smit (South Africa)Todd Smith (Canada)John Soghigian (United States)Philippe Solano (France)Tatiana Sulesco (Moldova)Marc Thellier (France)Jos\u00e9 Tort (Uruguay)Simon Tragust (Germany)Tom\u00e1\u0161 Tyml (Czech Republic)Maarten Vanhove (Belgium)Isabelle Villena (France)Philippe Vincendeau (France)Petr Volf (Czech Republic)Dominique Ang\u00e8le Vuitton (France)Hai-Long Wang (China)Lesley Warner Marion Wassermann (Germany)Richard Wilkerson (United States)Thierry Wirth (France)Samanta Xavier (Brazil)Masahiro Yamamamoto (Japan)Tingbao Yang (China)Yurong Yang Aneta Yoneva (Bulgaria)Hak Sun Yu (Korea)El\u017cbieta \u017bbikowska (Poland)Galina Zemstova (United States)Longxian Zhang (China)Nian-Zhang Zhang (China)Wenbao Zhang (China)Hejun Zhou (China)"} +{"text": "In the editorial section, Melanie M Taylor et al. (610) describe how syphilis screening and treatment can be integrated with HIV services. Mary Malebranche et al. (611) propose measures to alleviate the difficulties faced by pregnant refugees. This month\u2019s focus of the news section is the region of Africa. Tatum Anderson (614\u2013615) reports on progress made in the provision of family planning services and Charles Shey Wiysonge explains to Fiona Fleck (616\u2013617) how the Cochrane Collaboration contributes to evidence-based health care. Wanyue Zhang et al. (658\u2013662) report the effect of payments to providers for syphilis screening.Kebede Deribe et al. (653\u2013657) detail integrated care for lymphatic filariasis and podoconiosis. Alane Izu et al. (618\u2013629) track hospitalizations of children for pneumonia. A Pubudu De Silva et al. (647\u2013652) describe a reporting system to improve recall and follow-up.Sachiko Ozawa et al. (630\u2013639) estimate the economic impact of vaccinations programmes in 73 countries.Cara S Kosack et al. (640\u2013646) propose criteria for selecting diagnostic tests.Tsung-Ling Lee et al. (663\u2013664) argue for regulation of the stem cell industry."} +{"text": "International Health Regulations (2005).In the editorial section, Aziz Sheikh et al. (546) announce WHO's third Global patient safety challenge on reducing medication-related harm. Adilya Albetkova et al. (547) argue for specialized training of laboratory directors so that national governments can meet their obligations under the Sophie Cousins (550\u2013551) reports on the difficulties faced by national diabetes prevention and treatment programmes in WHO's South-East Asia region. Andr\u00e9ia Azevedo Soares (552\u2013553) interviews Sally J Rogers about evidence-based approaches to young children with autism.Shengjie Lai et al. (564\u2013573) analyse malaria trends and programme costs from 2011-2015.Jennifer Ho et al. (584\u2013593) collate the evidence for effectiveness of decentralized care.Maureen O'Leary et al. (574\u2013583) study low birth weight as a risk factor.Sabine Hermans et al. (554\u2013563) report a decrease in empirical treatment for tuberculosis. Viroj Tangcharoensathien et al. (599\u2013603) chart a path from global agenda to national health and agricultural actions.Pakoyo Fadhiru Kamba et al. (594\u2013598) assess the downsides of pharmaceutical donations.Carlos Dora et al. (604\u2013607) argue that environmental health policies can impact health outcomes. Steve W Lindsay et al. (606\u2013608) consider the spread of arboviruses and vector control measures."} +{"text": "In the editorial section, Walter Johnson et al. (634) explain why a global response is needed to prevent and treat stroke. Bernadette Abela-Ridder et al. (635) announce a new rabies vaccine stockpile. Gary Humphreys and Fiona Fleck (638\u2013639) report on efforts to address antimicrobial resistance, from national programmes to the United Nations General Assembly. Malcolm Molyneux tells Fiona Fleck (640\u2013641) about the challenges countries face with the first introduction of a malaria vaccine. Charles M Katz et al. (660\u2013666) model the interaction between imprisonment of gang members and homicide rates.Sergio Latorre-Arteaga et al. (652\u2013659) train teachers in rural communities to test children\u2019s vision.Cavin Epie Bekolo et al. (667\u2013674) analyse vaccine use, disease severity and deaths during an outbreak.Kirkby D Tickell and Donna M Denno (642\u2013651) review WHO guidelines on severe acute malnutrition.Mary Kay Kindhauser et al. (675\u2013686) establish the historical record of Zika virus prior to the current epidemic.Tim Crocker-Buque and Sandra Mounier-Jack (687\u2013693) gather stakeholders\u2019 perspectives of a finance facility. Lois Orton et al. (694\u2013704) review the evidence for health impacts of microfinance schemes.Nirmal Kandel et al. (705\u2013708) propose using polio surveillance systems for Zika-associated Guillain-Barr\u00e9 syndrome."} +{"text": "COMT Val158Met Genotype Determines the Direction of Cognitive Effects Produced by Catechol-O-Methyltransferase Inhibition\u201d by Farrell et al. . Specifically, in An error in"} +{"text": "In the editorial section, Christopher Dye and Shambhu Acharya (666) announce a theme issue on health in the sustainable development goals. Alarcos Cieza et al. (667) announce an upcoming theme issue on programmes to detect, prevent and treat visual impairments from all causes.In the news section, Tatum Anderson (670\u2013671) reports on efforts to train public health professionals in effective responses to false information about vaccines. Tabar\u00e9 V\u00e1zquez (672\u2013673), president of Uruguay, describes health reforms taken to reduce noncommunicable diseases.Janet Hoek et al. (726\u2013728) encourage more countries to insist on plain packaging for cigarettes.Carolina Nazzal and Jeffrey E. Harris (674\u2013682) track parallel trends in smoking bans and cardiovascular outcomes.Ravi Prakash Upadhyay et al. (706\u2013717) establish prevalence estimates for postpartum depression. Isabella Maina et al. (683\u2013694) use health facility data to assess subnational coverage of services.Bhola R Shrestha et al. (718\u2013719) argue for an effective antivenom.Manuel W Hetzel et al. (695\u2013705) track a decrease in malaria prevalence from 2008 to 2014.Anne Marie Thow et al. (723\u2013725) describe an import ban on turkey tails.Alessandra Ferrario et al. (720\u2013722) describe national collaboration on medicines procurement."} +{"text": "Unfortunately, after publication of this article (Chahla et al."} +{"text": "Dear editor,As corresponding author I ask for retraction of our article Finelli et al. (2016[Carmine Finelli, MD PhD"} +{"text": "Margaret E Kruk et al. (390) introduce this special issue on why and how to measure the quality of health-care provision., Amy Guthrie (393\u2013394) and Fiona Fleck report on efforts made in Mexico to improve services offered to people with diabetes. Catherine Calderwood (395\u2013396) explains how realistic medicine is being used to question variations in health outcomes across Scotland. In the news sectionLisa Marie Knowlton et al. (437\u2013444) compile a geospatial analysis of access to basic surgery. Claudia Hanson et al. (453\u2013464) analyse data from trials of community-based care provision.Barbara Madaj et al. (445\u2013452) propose indicators for quality of maternal and newborn care.Zhan Wang et al. (430\u2013436) explore the use of malpractice litigation records as an indication of problematic quality.Maureen E Canavan et al. (473\u2013477) study the introduction of quality measures. Gaurav Sharma et al. (419\u2013429) observe the care given to women during labour and delivery. Margaret E Kruk et al. (408\u2013418) document variation in the quality of care provided to mothers and babies.Marzia Lazzerini et al. (397\u2013407) do a randomized trial of staff supervision after training paediatricians in 20 hospitals. Yoko Akachi & Margaret E Kruk (465\u2013472) argue that patients\u2019 perceptions of care can be used to improve health services.Peter J Pronovost et al. (478\u2013480) present five narratives that impede progress towards safer care."} +{"text": "In this article \u2018 Serum Uric Acid and Mortality Form Cardiovascular Disease: EPOCH-JAPAN Study\u2019 by WenZhang, et al., which appeared in JAT 2016; 23: 692-703 are incorrect."} +{"text": "Unfortunately, the original article (Huang et al."} +{"text": "In the editorial section, Carla AbouZahr et al. (226) introduce an upcoming series on civil registration and vital statistics. Tatum Anderson (229\u2013230) reports on moves to improve cancer care provision in Botswana, Kenya and Rwanda. Fiona Fleck (231\u2013232) interviews Randall Packard about the lessons of global health histories. Lijian Han et al. (233\u2013242) estimate exposure to varied components of air pollution.Fentie Ambaw et al. (243\u2013255) study impacts on treatment outcomes, disability, and quality of life. Monique van Lettow et al. (256\u2013265) analyse missteps in HIV prevention programmes.Sophie Goyet et al. (286\u2013291) describe support provided to district health services.Christophe Van Dijck et al. (266\u2013280) review the evidence for antibiotic stewardship interventions.Alana Margaret Officer and V\u00e2nia de la Fuente N\u00fa\u00f1ez (295\u2013296) describe the effects of stereotypes used to describe older people.Hye-Na Kang and Ivana Knezevic (281\u2013285) detail evaluation methods needed to ensure safe and effective products. Dennis Carroll et al. (292\u2013294) introduce the global virome project."} +{"text": "The editorial team is grateful to all the experts who dedicated their time to peer-review our articles over the past 12 months. We wish to thank them and all those whose names are not listed below but gave helpful advice and support. We apologise to any reviewers whose names we may have missed.Our reviewers in 2016 were:Preben Aavitsland; Rachid Aaziz; Florence Abravanel; Cornelia Adlhoch; Franz Allerberger; Maria an der Heiden; S\u00f6ren Andersson; Marta Andr\u00e9s Miguel; Nick Andrews; Fabrizio Anniballi; Ozgur Araz; Maris Arcilla; Albena Arnaudova; Barry Atkinson; Jean-Luc Bailly; Meghan Baker; Koye Balogun; Elizabeth Bancroft; Norbert Bannert; Anne-Laure Ba\u00f1uls; Ian Barr; Nicholas Beeching; Edward Belongia; Peter Ben Embarek; Anne Berger-Carbonne; Bj\u00f6rn Berglund; Vincent B\u00e9ringue; Olaf Berke; Kathy Bernard; Antoine Berry; Zeno Bisoffi; Par Bjelkmar; Pierre-Yves Boelle; Shelly Bolotin; Magnus Boman; Francesco Bonfante; Isabelle Bonmarin; Marc Bonten; Stefan Borgmann; Scott Bowden; Walter Boyce; Heather Bradley; Marieta Braks; Arne Brants\u00e6ter; Viviane Bremer; Sharon Brookes; Tim Brooks; Lisa Brouwers; Kevin Brown; Mathias Bruyand; Mia Brytting; Udo Buchholz; Silke Buda; Herbert Budka; Daniel Cadar; Shona Cairns; Mattia Calzolari; Romina Camilli; Helen Campbell; Rafael Canton; Jo\u00e3o Andr\u00e9 Carri\u00e7o; Joan Casey; Alessandro Cassini; Nadim Cassir; Jes\u00fas Castilla; Dominique Caugant; Vicki Chalker; Meera Chand; Yves Charpak; Remi Charrel; Daniel Chemtob; Mark Chen; Allen Cheng; Gerardo Chowell; Bruno Ciancio; Niel Constantine; Victor Corman; Benjamin Cowling; Peter Coyle; Natasha Crowcroft; Maria Teresa Cuevas; Fran\u00e7ois Dabis; Harry Dalton; Stephanie Dancer; Niklas Danielsson; Mark Danta; Nadav Davidovitch; Helena de Carvalho Gomes; Birgitta de Jong; Jes\u00fas de Pedro Cuesta; Koen De Schrijver; Gaston De Serres; Henriette De Valk; Anouk Decors; Tarik Derrough; Jean-Claude Desenclos; Liselotte Diaz H\u00f6gberg; Sabine Diedrich; Mercedes Diez; David Dowdy; Jan Felix Drexler; Christian Drosten; Tarun Dua; Mariette F. Ducatez; Myrielle Dupont-Rouzeyrol; Boris Dzelalija; Paul Effler; Alex Ensminger; Susanna Esposito; Steen Ethelberg; Iro Evlampidou; David Eyre; Daniel Faensen; Dennis Falzon; Amanda Faulkner; Helmut Fickenscher; Nigel Field; James Fielding; Daniel Fierer; Antonietta Filia; Thea Fischer; David Fisman; Ross Fitzgerald; Julia Fitzner; Brendan Flannery; Ivo Foppa; Claudia Fortuna; Christina Frank; Eelco Franz; Derek Freedman; Alexander Friedrich; Ingrid Friesema; Hans-Peter Fuehrer; Katie Fullerton; Cristina Furtado; Carlo Gagliotti; Montserrat G\u00e1llego; Manuel Garc\u00eda Cenoz; Luis Garc\u00eda-Comas; Giuliano Garofolo; Anthony Gatt; Philippe Gautret; Peter Gerner-Smidt; Johan Giesecke; Pere Godoy; Magnus Gottfredsson; Stephan G\u00f6ttig; Hannelore Gotz; Luigi Gradoni; Kathie Grant; Donato Greco; Michel Grignon; Olaf Gudlaugsson; Volker Gurtler; Brian Gushulak; Jean-Paul Guthmann; Bernardo Rafael Guzman Herrador; Karin Haar; Walter Haas; Christos Hadjichristodoulou; Susan Hahne; \u00c1gnes Hajdu; Sebastian Haller; Pia Hardelid; Timm Harder; Heli Harvala; Henrik Hasman; Angelos Hatzakis; Christopher D. Heaney; Edou R. Heddema; Wiebke Hellenbrand; Greet Hendrickx; Bjorn Herrmann; David Heymann; Markus Hilty; Martin Hoenigl; Christiane H\u00f6ller; Emma Hollis; Vahur Hollo; Anders Holst; Vivian Hope; Judit Krisztina Horvath; G Hr\u010dkova; Yhu-Chering Huang; Judith H\u00fcbschen; Yvan Hutin; Benedikt Huttner; Daniela Huzly; Tetsuro Ikegami; Silviu Ionescu; Giuseppe Ippolito; Masako Iwanaga; Aftab Jasir; Johanna Jefferies; Cecilia Jernberg; Ana Jeroncic; Kari Johansen; Joerg Jores; Aleksandra Jovanovic Galovic; Dennis Junqueira; Bernard Kaic; Rolf Kaiser; Anu Kantele; Olen M Kew; Philip Kiely; Lisa King; Martyn Kirk; Esther Kissling; Irena Klavs; Silke Klee; Don Klinkenberg; Hilde Kl\u00f8vstad; Jan Kluytmans; Mira Kojouharova; Saara Kotila; Natalia Kozak-Muiznieks; Alenka Kraigher; Piotr Kramarz; Gerard Krause; Tyra Krause; Peter Kreidl; Karen A. Krogfelt; Adam Kucharski; Ed Kuijper; Martin Kulldorff; Jason C. Kwong; Guy La Ruche; Angie Lackenby; Shamez Ladhani; Philippe Lagac\u00e9-Wiens; Theresa Lamagni; Miranda Langendam; Karine Laroucau; Daniel Lavanchy; Jeffrey Lazarus; Simon Le Hello; Jason J Leblanc; Isabelle Leparc-Goffart; Isabel Leroux Roels; Justin Lessler; Daniel L\u00e9vy-Bruhl; S\u00e9bastien Lhomme; Troels Lillebaek; Bruno Lina; Marc Lipsitch; Irina Ljungqvist; Pierluigi Lopalco; Maria Angeles Lopaz; Rosa Lopez-Gigosos; Rachel Lowe; Claudia Lucarelli; Alexander Lukashev; \u00c5ke Lundkvist; Jennifer MacLachlan; Jean-Yves Madec; Gkikas Magiorkinis; Henri-Pierre Mallet; Helena Maltezou; Caterina Mammina; Jean-Michel Mansuy; Jean-Claude Manuguerra; Ulrich Marcus; Clara Mar\u00edn; Henju Marjuki; Laurence Marrama; Emily Martin; Susan Maslanka; Jean Mbisa; Peter McIntyre; Jim McMenamin; Mary Meehan; Adam Meijer; Angeliki Melidou; Ella Mendelson; Stefano Merler; Kevin Messacar; Elisabeth Meyer; Peter Michielsen; Nkuchia M'ikanatha; Vivi Miriagou; K\u00e5re M\u00f8lbak; Thomas Mollet; Carmen Monta\u00f1o-Remacha; Jacob Moran-Gilad; Simon More; Alain Moren; Robert Moss; Jo\u00ebl Mossong; Varvara Mouchtouri; Lapo Mughini-Gras; Miguel Mulders; Matthew Muller; Marcel M\u00fcller; Didier Musso; Thierry Naas; Celine A Nadon; Monika Naus; Richard Neher; Matthias Niedrig; Sandra Niendorf; Harold Noel; Paulo Jorge Nogueira; Hanna Nohynek; Hanne Nokleby; Teymur Noori; Daan Notermans; Pierre Nouvellet; Norbert Nowotny; Elina Numminen; Sophie Octavia; Ruth Offergeld; Justin O'Hagan; \u00c5ke \u00d6rtqvist; Masaki Ota; Makoto Ozawa; Takis Panagiotopoulos; Marcus Panning; Anna Papa; Vassiliki Papaevangelou; Dimitrios Paraskevis; Antonio Parisi; Gabriel Parra; Joseph Peiris; Luisa Peixe; Tatjana Pekmezovic; Pasi Penttinen; Kiran Perkins; Lyle Petersen; Efi Petinaki; Helen Petousis-harris; Martin Petric; G\u00fcnter Pfaff; Dina Pfeifer; Yvonne Pfeifer; Anastasia Pharris; Mathieu Picardeau; Denis Pi\u00e9rard; Johann Pitout; Diamantis Plachouras; Laurent Poirel; Michel-Robert Popoff; Thomas Pottage; Kevin Pottie; Edoardo Pozio; Anne Presanis; Elisabeth Presterl; Andrea Pugliese; Wolfgang Rabsch; Jayna Raghwani; Mario Ramirez; Brian Raphael; Rod Ratcliff; Gili Regev-Yochav; Chantal Reusken; Giovanni Rezza; Anders Rhod Larsen; Thomas V. Riley; Shmuel Rishpon; Caterina Rizzo; Beatriz Rojo-Bezares; Constanze Rossmann; Paul Rota; Adam Roth; Maja Rupnik; Etienne Ruppe; Werner Ruppitsch; Kristi R\u00fc\u00fctel; Tomoya Saito; Juan-Carlos Saiz; Stefania Salmaso; Dan Salmon; Lamia Samad; \u00d6rjan Samuelsen; Anita Sands; Monica Sa\u00f1\u00e9 Schepisi; Hugo Sax; Gianpaolo Scalia-Tomba; gaia scavia; Robert Scharff; Aaron Scherer; Patricia Schlagenhauf; Clara Schlaich; Axel Schmidt; Jonas Schmidt-Chanasit; Henrik Carl Sch\u00f8nheyder; Isabelle Schuffenecker; Lavinia Schuler-Faccini; Richard Selik; Ettore Severi; David Shay; Lester Shulman; Andrew Simpson; Andreas Sing; Vitali Sintchenko; Caroline Six; Lars Skog; Robert Skov; Danuta Skowronski; Werner Slenczka; Pierre Smeesters; David Smith; Heidi Soeters; Ruben Solano; Vincent Soriano; Mark Sotir; Erica Spackman; Gianfranco Spiteri; Danica Stanekova; Paola Stefanelli; Chen Stein-Zamir; Roger Stephan; Marc Struelens; Johan Struwe; Bertrand Sudre; Philippe Sudre; Carl Suetens; Jonathan Suk; Sheena Sullivan; Martin Sundqvist; Edit Szegedi; Silvio Tafuri; Johanna Takkinen; Yi-Wei Tang; Dennis Tappe; Masato Tashiro; Anders Tegnell; Rhys Thomas; Lelia Thornton; John Threlfall; Sera Tort; Katy Town; Stephen Tristram; Sotirios Tsiodras; Marta Valenciano; Janko van Beek; Birgit van Benthem; Wim van Bortel; Marita van de Laar; Marianne van der Sande; Marieke J. van der Werf; Steven Van Gucht; Albert van Hoek; Maria Van Kerkhove; Alies van Lier; Wilfrid Van Pelt; Ivo van Walle; Carmen Varela Santos; Istvan Varkonyi; Alkiviadis Vatopoulos; Inga Velicko; Harry Vennema; Jan Vinj\u00e9; Anne-Catherine Viso; Tanja Vollmer; Christina von Hunolstein; Rengina Vorou; Jiri Wagenaar; David Walker; Timothy Walker; Anders Wallensten; Jacco Wallinga; Hester Ward; Bryna Warshawsky; Pierre Wattiau; Todd Weber; Geoffrey Weinberg; Guido Werner; Katarina Westling; Ole Wichmann; Micael Widerstr\u00f6m; Miriam Wiese-Posselt; Hendrik Wilking; Christopher Williams; Carl-Heinz Wirsing von K\u00f6nig; James Wood; Hui-Ling Yen; Maria Zambon; Herve Zeller; Loukia Zerva; Ruth Zimmermann; Walter Zingg."} +{"text": "In the editorial section, Michelle J Hindin & Amanda M Kalamar (166) introduce new data sources indicating the contraceptive needs of adolescents.Etienne Krug & Alarcos Cieza (167) make a case for integrated rehabilitation services.Sophie Cousins (170\u2013171) reports on efforts in Sri Lanka to maintain unprecedented reductions in malaria transmission. Vikram Patel (172\u2013173) explains how people with depression can be treated in settings with no mental health professionals.Carolyn Staines & Joan Ozanne-Smith (174\u2013181) track a long-term reduction in deaths from drowning.Anna D Gage et al. (182\u2013190) study an approach to diagnosing performance. Thomas Jaenisch et al. (191\u2013198) estimate the risk of babies having microcephaly. Christine \u00c5rdal et al. (220\u2013226) weigh effective antibiotic stewardship against decreased commercial incentive. Krishna P Paudel et al. (227\u2013232) explore the feasibility of delivering other vaccines. Ebiowei SF Orubu & Catherine Tuleu (238\u2013240) explain why flexible solid oral formulations need to be readily available.Martyn D Kirk et al. (233\u2013234) estimate the paediatric disease burden due to contaminated food.Erin Sparrow et al. (235\u2013237) review potential uses for therapeutic antibodies.Jocelyn A Kessels et al. (210\u2013219) advocate for pre-exposure prophylaxis in high-risk populations.Aparna Schweitzer et al. (199\u2013209) analyse timing of Hepatitis B vaccination across 47 countries."} +{"text": "Darren B Taichman et al. (482\u2013483) present the International Committee of Medical Journal Editors\u2019 requirements for data-sharing statements when reporting clinical trials. Timothy G Evans and Marie Paule Kieny (484) explain how health systems research is needed to inform progress towards universal health coverage.In the news section, Apiradee Treerutkuarkul (487\u2013488) reports on policies in Thailand to prevent alcohol-related harm. Frederico C Guanais tells Andr\u00e9ia Azevedo Soares (489\u2013490) how patient empowerment can lead to improvements in health \u2013care quality.Guadalupe Bedoya et al. (503\u2013517) study compliance with infection prevention and control practices.Nana Mensah Abrampah et al. (527\u2013531) emphazise the need for sanitation in health care facilities. Stephan Brenner et al. (491\u2013502) study an incentive scheme to address shortages of medical supplies and to improve use of clinical protocols.Mark JD Jordans et al. (532\u2013536) propose ways to improve case-finding.Hsiao-Han Chang et al. (518\u2013526) examine genetic data for use in the design of Zika diagnostics.Marie Paule Kieny et al. (537\u2013539) link sustainable development to universal health coverage. Nick Wilson et al. (540\u2013541) list the pros and cons of banning electronic cigarettes. Brooke Ronald Johnson Jr et al. (542\u2013544) introduce a new database of national abortion laws."} +{"text": "In the editorial section, Patrick L Osewe (794) discusses options for financing pandemic preparedness. Dermot Maher & Nathan Ford (795) propose that the public health research agenda is informed by questions arising during guideline development. Tatum Anderson (798\u2013799) reports on measures taken by several countries to prevent skin cancer by restricting the use of sunbeds. Fatima Suleman talks to Fiona Fleck (800\u2013801) about efforts to make affordable medicines available, particularly in South Africa. Ralph Huits et al. (802\u2013810) quantify Zika virus in semen samples.Asa Auta et al. (831\u2013841) collate the evidence on health care workers\u2019 exposures to body fluids. Praveen Gadde et al. (848\u2013849) chart a difficult course of disease definition and prevention.Stania Kamara et al. (842\u2013847) describe mental health care provided during the Ebola outbreak. Claudia Hanson et al. (811\u2013821) document geographical inequalities in access to services.Lirije Hyseni et al. (822\u2013830) review the evidence for interventions designed to reduce trans-fat.Catherine Jan et al. (850\u2013852) discuss health trends of migrant workers\u2019 children."} +{"text": "In editorials, Monica Di Luca et al. (298) argue for earlier diagnosis and treatment of disorders of the brain. Duane J Gubler (299) describes the implications of further spread of yellow fever.Sam Loewenberg (302\u2013303) reports on the challenges faced in Zambia\u2019s attempt to eliminate malaria. Yogan Pillay talks to Fiona Fleck (304\u2013305) about South Africa\u2019s moves to integrate service provision for tuberculosis and HIV.Cheryl J Cherpitel et al. (335\u2013342) test the validity of coding for a single episode of alcohol use resulting in injury.Lijun Wang et al. (314\u2013326) track deaths from poisoning by intent and substance from 2006 to 2016.Haruka Sakamoto et al. (355\u2013359) describe efforts to influence policy at meetings between governments of Canada, France, Germany, Italy, Japan, the United Kingdom of Great Britain and Northern Ireland and the United States of America.Juan Eugenio Hern\u00e1ndez-\u00c1vila et al. (306\u2013313) estimate Zika virus infections.K LeRoux et al. (360\u2013365) report lessons learnt in a rabies control programme.Taryn Silver Lorthe et al. (327\u2013334) evaluate an outbreak verification system.Shannon E Brent et al. (343\u2013354) examine the potential for urban introduction of yellow fever virus by international travellers.150Andrew M Briggs et al. (366\u2013368) call for action to reduce the global burden of musculoskeletal conditions."} +{"text": "The editorial team is grateful to all the experts who dedicated their time to peer-review our articles over the past 12 months. We wish to thank them and all those whose names are not listed below but gave helpful advice and support. We apologise to any reviewers whose names we may have missed.Our reviewers in 2017 were:Frank Aarestrup; Preben Aavitsland; Muna Abu Sin; Ibrahim Abubakar; Natalie Adams; Cornelia Adlhoch; Jan Albert; Barbara Albiger; Franz Allerberger; Gayatri Amirthalingam; Nick Andrews; Denise Antona; Katherine Atkinson; Sabrina Bacci; Michael Baker; Elizabeth Bancroft; Francis Barin; Ian Barr; Aldert Bart; Cornelius Bartels; Luisa Barzon; Leonardo Basco; Anna Baumann-Popczyk; Frank Beard; Angela Bechini; Ermias Belay; Rutger Bennet; Kimberley Benschop; Anne Berger-Carbonne; Silvia Bertagnolio; Marianne Besnard; Rijkelt Beumer; Philippe Beutels; Beverly Biggs; Zeno Bisoffi; Carina Blackmore; Jesse Blanton; Pierre-Yves Boelle; Shelly Bolotin; Magnus Boman; Marc Bonten; Thijs Bosch; Arnold Bosman; Herve Bourhy; Virginia Bowen; Karoline Bragstad; Christian Brandt; Andrew Breed; Viviane Bremer; Eeva Broberg; Udo Buchholz; Jane Buxton; Stefan B\u00f6rjesson; Sindy B\u00f6ttcher; Saverio Caini; Helen Campbell; Christine Campese; Yehuda Carmeli; Jo\u00e3o Andr\u00e9 Carri\u00e7o; Jordi Casabona; Jean-Sebastien Casalegno; Francesco Castelli; Jesus Castilla Catalan; Dominique Caugant; Rachel Chalmers; Mark Chen; Stephane Chevaliez; Gerardo Chowell; Peer Brehm Christensen; Abrar Chughtai; Konstantin Chumakov; Nadia Ciampa; Bruno Ciancio; Vittoria Colizza; Simon Collin; Geoffrey Coombs; Helena Cortes Martins; Laurent Cotte; Suzanne Cotter; Anthony Cousien; Susan Cowan; Benjamin Cowling; Ingmar Cox; Adam Crawley; Natasha Crowcroft; Ida Czumbel; Viktor Dahl; C\u00e9dric Dananch\u00e9; Niklas Danielsson; Mark Danta; Siddhartha Sankar Datta; Jean-Philippe David; Nadav Davidovitch; Pieter de Boer; Jes\u00fas de Pedro Cuesta; Gaston De Serres; Rory D. de Vries; Sylvia Declich; Valerie Delpech; Tarik Derrough; Sarika Desai; Sergio Di Martino; Liselotte Diaz H\u00f6gberg; Sabine Diedrich; Mercedes Diez; Gerhard Dobler; Yohei Doi; Dragoslav Domanovic; Lisa Domegan; Raquel Duarte; Sandra Dudareva-Vizule; Erika Duffell; Erwin Duizer; Alexandre Duvignaud; Michael Edelstein; Ronald E Engle; Connie Erkens; Douglas Esposito; Steen Ethelberg; Matthieu Eveillard; Mirko Faber; Michael Farrell; Margaret Fearon; Jill Ferdinands; Laura Fern\u00e0ndez-L\u00f3pez; Antonietta Filia; William Fischer; Thea Fischer; Marc Fischer; David Fisman; Claude Flamand; Brendan Flannery; Laure Fonteneau; Frode Forland; Ron Fouchier; Pieter Fraaij; Ellen Fragaszy; Christina Frank; Eelco Franz; Joanne Freedman; Alexander Friedrich; Ingrid Friesema; Angelika Fruth; Alicia Fry; Giovanni Gabutti; Kartini Gadroen; Monica Galiano; Lauren Gardner; Patricia Garvey; Maja George; Peter Gerner-Smidt; Giovanni Gherardi; Eleni Giamarellou; Monica Gianfranceschi; Heather Gidding; Nicolas Gilbert; Vladimir Gilca; Enrico Girardi; Anja Globig; Pere Godoy; Daniel Golparian; Claire Gordon; Marga Goris; Magnus Gottfredsson; Luigi Gradoni; Tine Grammens; Kathie Grant; Stephen Graves; Michel Grignon; Diane Gross; Andrew Grulich; Beatriz Guerra; Barbara Gunsenheimer-Bartmeyer; Jean-Paul Guthmann; Huldrych G\u00fcnthard; Walter Haas; Josipa Habus; \u00c1gnes Hajdu; Irene Hall; Ian Hall; Anette Hammerum; Dag Harmsen; Michael Hartal; Heli Harvala; Henrik Hasman; Angelos Hatzakis; Barbara Hauer; Amber Haynes; Shirin Heidari; Franz Heinz; Wiebke Hellenbrand; Rene Hendriksen; Ursel Heudorf; Peter Hoffman; Sven Hoffner; Vahur Hollo; Judit Krisztina Horvath; Jaroslav Hrabak; Darko Hren; Gwenda Hughes; Benedikt Huttner; Judith H\u00fcbschen; Thomas Inns; Angela Iuliano; Jacques Izopet; Michael Jackson; Katri Jalava; Klaus Jansen; Adam Jenney; Kari Johansen; Nathalie Jourdan-da Silva; Laila Kahwati; Bernard Kaic; Rolf Kaiser; Tommi Karki; Margaret Kay; Brigitte Keller-Stanislawski; Kamran Khan; Yury Khudyakov; Pete Kinross; Hilary Kirkbride; Peter Kirwan; Esther Kissling; Anda Kivite; R. Monina Klevens; Hilde Kl\u00f8vstad; Gary Kobinger; Anke Kohlenberg; Frank Konings; Marion Koopmans; Michael Kosoy; Gerard Krause; Peter Kreidl; Karl G. Kristinsson; Paul Krogstad; Ed Kuijper; Jeffrey Kwong; Angie Lackenby; Shamez Ladhani; Kaja-Triin Laisaar; Marlyn Langford; Simone Lanini; Amparo Larrauri; Jesper Larsen; Guiseppe Latorre; Camille Lebarbenchon; Tinne Lernout; Jessica Leung; Alexandra Levitt; Tore Lier; Marshall Lightowlers; Giedrius Likatavicius; Jing Liu-Helmersson; Luis Lowe; Jay Lucidarme; Outi Lyytik\u00e4inen; Daniel L\u00e9vy-Bruhl; Knut L\u00f6nnroth; Ian Mackay; Fabio Magurano; Suman Majumdar; Helena Maltezou; Ula Maniewski; Jean-Michel Mansuy; Ulrich Marcus; Laurence Marrama; Kim Marsh; Rebecca Martin; Emily Martin; Valentina Marziano; Brian Maskery; Andrew McArthur; Jim McMenamin; Adam Meijer; Angeliki Melidou; Tanya Melillo Fenech; Alexander Mellmann; Kassiani Mellou; Stefano Merler; Tony Merritt; Thomas Mertens; Thomas Mollet; Dominique Monnet; Arnold Monto; Hannah Moore; Orna Mor; Jacob Moran-Gilad; Serge Morand; Alain Moren; Jane Morgan; Jo\u00ebl Mossong; Antons Mozalevskis; Viktor Mrav\u010d\u00edk; Lapo Mughini-Gras; Mark Muscat; James Musser; Didier Musso; Mette Myrmel; K\u00e5re M\u00f8lbak; Thomas M\u00fcller; Celine A Nadon; Andreas Neumayr; Emanuele Nicastri; Matthias Niedrig; Eva M\u00f8ller Nielsen; Sandra Niendorf; Hubert Niesters; Taina Niskanen; Harold Noel; Hanne Nokleby; Teymur Noori; Helene Norder; Baltazar Nunes; Pat Nuttall; Domingo Nu\u00f1ez; Darina O'Flanagan; Ruth Offergeld; Makoto Onishi; Laura Pabst; Daniel Palm; Takis Panagiotopoulos; Marcus Panning; Annalisa Pantosti; Dimitrios Paraskevis; Elena Pariani; R David Parker; Suzan Pas; Richard Pebody; Malik Peiris; Joseph Peiris; Luisa Peixe; Pasi Penttinen; Agn\u00e8s Perrin; Martin Petric; Mariska Petrignani; G\u00fcnter Pfaff; Dina Pfeifer; Yvonne Pfeifer; Anastasia Pharris; Denis Pierard; Sylvie Pillet; Rosa Pint\u00f3; Diamantis Plachouras; Laurent Poirel; Isabelle Poujol; Edoardo Pozio; Elisabeth Presterl; Jan Prins; Matthias Pulz; Mikkel Quam; Lul Raka; Brian Raphael; Lasse Dam Rasmussen; Helena Rebelo-de-Andrade; Claudia Reinheimer; Chantal Reusken; Giovanni Rezza; Anders Rhod Larsen; Caterina Rizzo; Emmanuel Robesyn; Mohammad Rokni; Roberto Romi; Karin Ronning; Anne-Marie Roque-Afonso; Senia Rosales-Klintz; Mirko Rossi; Gian Maria Rossolini; Omar Rota Stabelli; Adam Roth; Etienne Ruppe; Kate Russell; Malgorzata Sadkowska-Todys; Daniel Sagebiel; Mika Salminen; Lamia Samad; Jussi Sane; Andre Sasse; Rachel Savage; Aaron Scherer; Barbara Schimmer; Jonas Schmidt-Chanasit; Holger Scholz; Jennifer Schuster; Brunhilde Schweiger; Tara Sealy; Harald Seifert; Torsten Semmler; Helena Seth-Smith; Torsten Seuberlich; Ettore Severi; Otilia Sfetcu; Lester Shulman; Annette Siedler; Ian Simms; Andreas Sing; Vitali Sintchenko; Gunilla Skoog; Robert Skov; Ram\u00f3n Soriguer; Gianfranco Spiteri; Erin Staples; Cristina Stasi; Kim Steegen; Paola Stefanelli; Pawel Stefanoff; Marc Stegger; Chen Stein-Zamir; John Stenos; Alistair Story; Errol Strain; Franc Strle; Bertrand Sudre; Carl Suetens; Jonathan Suk; Sheena Sullivan; Edit Szegedi; Muhamed-Kheir Taha; Emi Takashita; Theresa Tam; Lara Tavoschi; Melanie M Taylor; Karin Tegmark-Wisell; Anders Tegnell; Daniel Thomas; Lelia Thornton; Karen Tiley; Elizabeth Torrone; Maria Elena Tosti; Mathieu Tourdjman; Katy Town; Sotirios Tsiodras; Nikki Turner; Bianca T\u00f6r\u00f6s; Bernhard Ultsch; Tim Uyeki; Palle Valentiner-Branth; Wim Van Bortel; Marita van de Laar; Kees. C. van den Wijngaard; Wim van der Hoek; Marianne van der Sande; Sylvie van der Werf; Marieke J. van der Werf; Gary van Domselaar; Johan Van Griesven; Albert Jan van Hoek; Esther van Kleef; Edward van Straten; Pieter van Thiel; Ivo van Walle; Carmen Varela Santos; Istvan Varkonyi; Alkiviadis Vatopoulos; Sophie Vaux; Irene Veldhuijzen; Harry Vennema; Giulietta Venturi; Line Vold; Gwena\u00ebl Vourc\u2019h; Sabine Vygen; Anders Wallensten; Crystal Watson; Pierre Wattiau; Nina Weis; Don Weiss; Dirk Werber; Guido Werner; Therese Westrell; Ole Wichmann; Katarina Widgren; Annelies Wilder-Smith; Hendrik Wilking; Kumanan Wilson; Carl-Heinz Wirsing von K\u00f6nig; Katja Wolthers; William WL Wong; Edwina Wright; Alban Ylli; Hans L Zaaijer; Katherina Zakikhany; Philipp Zanger; Gianluigi Zanusso; Peter Zarb; Gernot Zarfel; Herve Zeller; Ruth Zimmermann; Walter Zingg; Phillip Zucs; Milan \u010ci\u017eman; Barbara \u0160oba."} +{"text": "In the editorial section, Hajime Inoue & Ren Minghui (242) advocate for immediate national action on antimicrobial resistance. Katherine Littler et al. (243) review progress on data sharing in public health emergencies.Claire Keeton (246\u2013247) reports on how social innovation can improve access to health care in low-income countries in Africa. Gary Schwitzer (248\u2013249) explains to Fiona Fleck how he seeks to improve media coverage of health topics. Ashish KC et al. (261\u2013269) analyse immunization coverage and patterns from 2001\u20132014. William E Oswald et al. (250\u2013260) quantify associations between community sanitation use and children with active trachoma.Tom Wingfield et al. (270\u2013280) randomize households to test the effects of socioeconomic support.Amber Hsiao et al. (303\u2013312) describe 12 cholera campaigns in 9 countries.Hamish Graham et al. (288\u2013302) analyse what\u2019s needed to improve oxygen delivery to paediatric patients. Lana Clara Chikhungu et al. (281\u2013287) review the evidence for links between child and maternal mortality."} +{"text": "Unfortunately, after publication of this article (Bonnin et al."} +{"text": "In the editorial section, Piyasakol Sakolsatayadorn and Margaret Chan (86) introduce this special issue on the health of vulnerable populations. Abdul Ghaffar et al. (87) explain why health systems research needs to be done by the people who will use the results.Shiva Raj Mishra (90\u201391) reports on sexual and reproductive health services and education programmes for adolescents in Nepal.Fiona Fleck interviews Mani Jegathesan (92\u201393) about his experience as an Olympic athlete and his subsequent career as an advocate for public health.Tefera Darge Delbiso et al. (94\u2013102) study the associations between drought and conflict and childhood wasting and stunting.Leonor Maria Pacheco Santos et al. (103\u2013112) observe the effects of policies to deploy more doctors.Peter W Luckow et al. (113\u2013120) examine ways of delivering maternal and child health servicesDipu Joshi et al. (135\u2013139) trial case-finding by peers of people living with HIV.Viroj Tangcharoensathien et al. (146\u2013151) assess the introduction of health insurance for migrant workers.Sirinart Tongsiri et al. (140\u2013145) detail efforts to adapt home environments for people with disabilities.Catherine Arsenault et al. (128\u2013134) create a dashboard to monitor vaccination coverage.Daniela C Rodr\u00edguez et al. (121\u2013127) argue for ongoing political commitment.Rebekah Thomas et al. (154\u2013156) list steps to ensuring the inclusion of transgender people.Emmanuel d'Harcourt et al. (157\u2013158) consider how to improve the lives of people affected by conflict.Jody-Anne Mills et al. (162\u2013164) explain why rehabilitation services should be part of disaster response.Devaki Nambiar & Harsh Mander (152\u2013153) enumerate health risks faced by poor people in urban settings.Matthew Grenall et al. (159\u2013161) follow an evolving focus on vulnerable populations by the Global Fund to Fight AIDS, Tuberculosis and Malaria."} +{"text": "Nabil Karah et al. (370) call for better provision of dialysis services to Syrian refugees in Lebanon. Maggie Montgomery et al. (371) point out that cholera prevention requires adequate water and sanitation in all settings.Tatum Anderson (374\u2013375) reports on the need for a global response to an number of increased yellow fever outbreaks. Dixon Chibanda explains to Fiona Fleck (376\u2013377) how he developed the Friendship Bench approach to mental health care in Zimbabwe. Giorgia Sulis et al. (386\u2013392) implement prevention for exposed children.Maurice M\u2019bangombe et al. (428\u2013435) examine the use of oral cholera vaccine.Abdulgafoor M Bachani et al. (423\u2013427) present the results of a postgraduate training course.Lukas Roth et al. (378\u2013385) identify gaps in public pharmaceutical standards.Mark G Shrime et al. (393\u2013401) present a data envelopment analysis adapted to the health system. Michelle M Haby et al. (402\u2013413) review evidence for prevalence estimates of asymptomatic infection.Valerie A Luyckx et al. (414\u2013422) show how all goals can be connected to kidney disease prevention and treatment. Julian Fisher et al. (436\u2013438) outline steps towards the phase down of dental amalgam.Bettina Borisch et al. (439\u2013440) provide an update on the Global Charter for the Public\u2019s Health."} +{"text": "Sir,et al[triphala and chlorhexidine mouthwashes.[in vitro effect on drug modulating enzymes.[I read with great interest a recent published work on triphala mouthwash by Tandon et al They conthwashes. I agree thwashes. However, enzymes. Disturba enzymes."} +{"text": "Standardized incidence ratios show significant overall excesses in south Asians (131), largely due to higher rates in south Asian boys, and specific excesses for leukaemia (141), lymphoid leukaemia (141), lymphoma (172) and hepatic tumours (375). Aetiological investigation is required. \u00a9 2001 Cancer Research Campaign"} +{"text": "Sir,et al.[et al. finally suggested an association of DD polymorphism and type II diabetes with nephropathy.\"[I read with great interest the recent report by Naresh et al. Naresh eropathy.\" Indeed, ropathy.\" However,ropathy.\" This datropathy.\""} +{"text": "Organotin(IV) complexes were more active than tin(II) complexes.Some organotin(IV) and tin(II) complexes of composition R"} +{"text": "Other therapeutic options being explored for CP/CPPS include tricyclics, anticonvulsants, physical therapy, and myofascial or trigger point release.[The most common urologic diagnosis made in men younger than 50 years is prostatitis. Till dat3et al. were adel.[et al. was thatl.[et al. alfuzosi release. The resu"} +{"text": "Sir,et al. on needle stick among healthcare workers (HCWs).[et al. concluded that \u201cHCWs at Aleppo University hospitals are frequently exposed to blood-borne infections. Precautions to avoid, and protection from, needle stick infections (NSIs) are important in preventing infection of HCWs. Education about the transmission of blood-borne infections, vaccination and post-exposure prophylaxis must be implemented and strictly monitored.\u201d[I read with great interest the recent publication by Yacoub s (HCWs). Yacoub enitored.\u201d Indeed, nitored.\u201d Hence imnitored.\u201det al.[There might have been important technical problems in the study by Yacoub et al."} +{"text": "In the editorial section, Cornelia Anne Barth (790) outlines steps needed to provide better care for 20 million people with physical disabilities living in crisis settings. Onyema Ajuebor et al. (791) introduce a curricula guide for the training of health workers in antimicrobial resistance prevention measures. Ritu Sadana et al. (792) outline the evidence needed to track the health of ageing populations.In the news section, Andrey Shukshin and Gary Humphreys (795\u2013796) report on drug resistant tuberculosis control programmes in Belarus. Gary Humphreys interviews Ahd Hamidi (797\u2013798) about her work in technology transfer for vaccines. Angela Devine et al. (828\u2013836) use primary trial data to extract costs per episode.Manisha Dubey et al. (799\u2013809) analyse the classification effects of treatment guidelines. Irene Njuguna et al. (837\u2013845) propose ways to improve the transition from pediatric to adult HIV care.Kawther M Hashem et al. (818\u2013827) survey caloric contents of carbonated sugar-sweetened beverages. Cathy Green et al. (810\u2013817) mobilize the community to improve outcomes. Pepita Barlow et al. (846\u2013848) study the interactions between the World Health Organization and World Trade Organization.Joseph Wong & Kimberly Skead (849\u2013850) examine cost estimates of universal coverage.Kate Elder et al. (851\u2013853) describe a humanitarian mechanism for pneumococcal vaccines.Lawson Ung et al. (854\u2013856) argue that infectious corneal ulceration should be considered a neglected tropical disease."} +{"text": "In the editorial section this month, David Watkins (442) considers policy options for diet-related noncommunicable diseases. Peter Salama et al. (443) discuss the implications of research on health-worker strikes in low-countries.Andrey Shukshin (446\u2013447) reports on progress made in the Russian Federation to improve the treatment of childhood cancer. Sandro Galea talks to Gary Humphreys (448\u2013449) about the power of epidemiology and the need to change the way we talk about health.Tarsicio Uribe-Leitz et al. (501\u2013512) examine trends of caesarean delivery.Francisco Goiana-da-Silva et al. (450\u2013459) model impacts of food industry co-regulation on noncommunicable diseases.Nadine Nannan et al. (468\u2013476) estimate completeness of birth registration.Meghan A Bohren et al. (477\u2013484) measure gender balance in WHO panels that develop guidelines.Giuliano Russo et al. (460\u2013467) gather the available evidence on health workers\u2019 strikes in low-income countries.Yanhong Jessika Hu et al. (485\u2013500) review antibiotic resistance data from infections in children in 20 countries."} +{"text": "In editorials this month, Diana Zandi et al. (2) call for papers on the subject of artificial intelligence in the health sector. Wenjing Tao et al. (3) discuss the proxy indicators used to estimate antibiotic consumption by people and the continued surveillance needed to control antimicrobial resistance.Convention on the Rights of Persons with Disabilities.Tatum Anderson and Gary Humphreys (6\u20137) report on recent outbreaks and global spread of Zika virus disease. Catalina Devandas Aguilar talks to Stephanie Cheng (8\u20139) about the impact of the 2008 Britt McKinnon & Ashley Vandermorris (42\u201350) query the associations between age-of-consent to testing and knowledge of HIV status. Yuehong Wei et al. (51\u201358) analyse decades of rabies control efforts.Nancy Armenta Paulino et al. (59\u201367) examine inequities in the provision of maternal health care. Paula Braitstein et al. (33\u201341) measure HIV prevalence in young people and children living on the streets.Samuel Bawa et al. (24\u201332) study the feasibility of delivering primary health care instead of polio vaccines. Angela Y Chang et al. (10\u201323) track how people fare when they have other chronic diseases."} +{"text": "New tick-borne encephalitis virus hot spot in Northern Zealand, Denmark, October 2019\u2019 by CN Agergaard et al., published on 24 October 2019, GenBank accession numbers were added on 4 December 2019.In the article \u2018"} +{"text": "In an editorial, Ab Fatah Ab Rahman (729) argues for increased capacity to monitor adverse drug reactions in community settings. In the news section, Sophie Cousins (733\u2013734) reports on urban programmes to reduce gun violence in Colombia and interviews Balram Bhargava (735\u2013736) about his efforts to foster needed innovation in the health sector.Tieba Millogo et al. (783\u2013788) examine the effects of task-shifting.Sue Napierala et al. (764\u2013776) study ways of providing self-testing kits.Ivan Meshkov et al. (737\u2013745) examine the variables affecting tuberculosis prevalence.Michael Edelstein et al. (754\u2013763) triangulate data sources.Bernadette Ann-Marie O\u2019Hare (746\u2013753) estimates the potential health sector gains of addressing international corporate tax avoidance.Neil Armstrong and Jo Welsman (777\u2013782) examine myths and misconceptions."} +{"text": "In the editorial section this month, Shamsuzzoha Babar Syed et al. (2) ask how quality of care can be improved in fragile, conflict-affected and vulnerable settings. The World Health Organization (3) issues a retraction statement on guidance for opioid use. In the news section Sophie Cousins (6\u20137) reports on health service outreach using hospital trains in China and India. Gary Humphreys interviews Kathryn Chu (8\u20139) on the intersection of surgery and public health and her research on improving the quality of surgical delivery in humanitarian crises. Daphne CN Wu et al. (19\u201329) review investments made for urban populations.Xiaochu Yu et al. (10\u201318) project per-capita blood supply and demand.Manas K Akmatov et al. (40\u201351) analyse the spatial distribution of outpatient claims for asthma treatment.Andrea Farnham et al. (69\u201371) argue for more use of district health information.Chunling Lu et al. (30\u201339) estimate amounts of development assistance provided to 114 countries.Kanna Sugiura et al. (52\u201358) examine the evolution of mental health care.Yot Teerawattananon et al. (59\u201365) discuss charging policies for survey instruments.Newton Opiyo et al. (66\u201368) present WHO recommendations for non-clinical interventions."} +{"text": "In the editorial section, Flavio Salio and Altaf Musani (310) call for more research on the delivery of health services to people living in conflict settings. Mercedes Tatay and Els Torreele (311) ask for financing mechanisms to ensure access to essential medicines when countries are no longer eligible for assistance from the Global Fund.WHO Global code of practice on the international recruitment of health personnel. Gary Humphreys (314\u2013315) reports on an initiative to improve emergency care in district hospitals in Uganda. Rosalinda Dimapilis-Baldoz talks to Fiona Fleck (316\u2013317) about the health worker migration policy of the Philippines and the Karl Blanchet et al. (374\u2013376) report on priority-setting measures. Selma Gicevic et al. (349\u2013357) propose a low-cost model for dietary surveillance. Szabolcs Szigeti et al. (335\u2013348) analyse revenue sources. Pradeep Haldar et al. (328\u2013334) recount lessons learnt from fractional dosing. Trishul Siddharthan et al. (318\u2013327) measure prevalence in urban and rural settings.Nanna Maal\u00f8e et al. (365\u2013370) adapt intrapartum care guidelines. David Beran et al. (358\u2013364) assess vulnerabilities to insulin shortages.Barclay Stewart et al. (371\u2013373) argue that organized trauma care needs a greater share of development aid."} +{"text": "In the editorial section, Robert Marten et al. (798) discuss a shift in global governance needed to reach the sustainable development goals. Shamsuzzoha B Sayeed et al. (799) describe national efforts to integrate health-care quality improvement initiatives.In the news section, Vijay Shankar Balakrishnan (802\u2013803) reports the challenges in preventing, detecting and treating hepatitis B and C infections in the European Region. Jean-Jacques Muyembe Tamfum talks to Fiona Fleck (804\u2013805) about his decades of experience in responding to Ebola virus disease outbreaks.Xin Wang et al. (843\u2013852) describe moves toward people-centred integrated care.Shohei Okamoto et al. (826\u2013833) estimate the health effects for men of working past their retirement age.Francesco Grandesso et al. (817\u2013825) document a cholera vaccination programme. Tina Lavin et al. (806\u2013816) classify perinatal mortality data.Netnapis Suchonwanich & Winai Wananukul (853\u2013857) document improved access to antidotes and antivenoms.Anna Hidle et al. (834\u2013842) count the costs of a human papillomavirus vaccination project.Graziella Iossa & Piran CL White (858\u2013860) identify a missing link in plans to combat antimicrobial resistance.Debra Jackson et al. (861\u2013863) explain how civil registration and vital statistics are used by health systems."} +{"text": "The editorial team is grateful to all the experts who dedicated their time to peer-review our articles over the past 12 months. We wish to thank them and all those whose names are not listed below but gave helpful advice and support. We apologise to any reviewers whose names we may have missed.Our reviewers in 2019 were:Sam Abbott; Stephan Aberle; Amos Adler; Cornelia Adlhoch; Karoline Aebi-Popp; Anton Aebischer; Seven Johannes Sam Aghdassi; Franz Allerberger; Erik Alm; Katharina Alpers; Christian Althaus; Laura Anderson; Nick Andrews; Andrea Angheben; Muna Anjum; Markus Antwerpen; Kevin Ari\u00ebn; Wouter Arrazola de O\u00f1ate; Mardjan Arvand; Thomas Atkinson; Agoritsa Baka; Vincenzo Baldo; Eszter Balla; April Baller; Mathieu Bangert; Fernardo Baquero; Ian Barr; Luisa Barzon; Ulrike Baum; Julien Beaut\u00e9; Ron Behrens; Antonino Bella; Edward Belongia; Kimberley Benschop; Dennis Bente; Bojana Beovi\u0107; B\u00e9atrice Ber\u00e7ot; Juan Berenguer; Christoph Berger; Claire Bertelli; Sophie Bertrand; Marianne Besnard; Debra Bessen; Cornelia Betsch; Yunus Emre Beyhan; Julia Bitzegeio; Carina Blackmore; Amandine Boeuf; Shelly Bolotin; Raquel Bonelli; Marc Bonten; Michael Borg; Jo\u00e3o Botelho; Herv\u00e9 Bourhy; Martijn Bouwknegt; Naomi Boxall; Marieta Braks; Viviane Bremer; Deborah Briggs; Folke Brinkmann; Eeva Broberg; Tal Brosh-Nissimov; Julia Brotherton; Colin Brown; Springer Browne; Mia Brytting; Jacqueline Bua; Silke Buda; Susanne buder; Niccol\u00f2 Buetti; Muruleedhara Byappanahalli; Sindy B\u00f6ttcher; Saverio Caini; Paolo Calistri; Christine Campese; Rosa Cano; Catherine Carillo; Yehuda Carmeli; AnnaSara Carnahan; Jo\u00e3o Andr\u00e9 Carri\u00e7o; Otto Cars; Jordi Casabona; Jean-Sebastien Casalegno; Alessandro Cassini; Francesco Castelli; Jesus Castilla Catalan; Vincent Cattoir; Dominique Caugant; Matthias Cavassini; Remi Charrel; Anuja Chatterjee; Didier Che; Daniel Chemtob; Yu Chen; Tom Chiller; Sadegh Chinikar; Gerardo Chowell; Iva Christova; Nadia Ciampa; Tristan Clark; John Coia; Rosalind Lucy Coleman; Vittoria Colizza; Sarah Collier; Deirdre Collins; Alex Cook; Michael Cooper; Vicente Corrales-Medina; Andrea Cortegiani; Benjamin Cowling; Amelieke Cremers; Sara Croxford; Gabriela da Silva; Ron Dagan; Eilif Dahl; Richard Danila; Eric Dannaoui; Mark Danta; Nicolas Dauby; Helena de Carvalho Gomes; Alexis de Rougemont; Gaston De Serres; Jaime Delgado; Valerie Delpech; Sarika Desai; Antonino Di Caro; Liselotte Diaz H\u00f6gberg; Julia Dina; Steven Djordjevic; Gerhard Dobler; Roger Dodd; Margaret Doll; Pierre Dorny; Jan Drexler; Christian Drosten; Matthew Dryden; Sandra Dudareva-Vizule; Erika Duffell; Tim Eckmanns; Adrian Egli; Anna-Maria Eis-Huebinger; Andreas Halgreen Eiset; Joanna Ellis; Theresa Enkirch; Olivier Epaulard; Connie Erkens; Sylvie Escolano; Steen Ethelberg; Matthieu Eveillard; Mirko Faber; Dennis Falzon; Cecilia Fazio; Vincenzo Ferrantelli; Lena Fiebig; Nigel Field; Helen Fifer; Marc Fischer; Thea Fischer; David Fisman; Brendan Flannery; Gilles Foll\u00e9a; Anders Fomsgaard; Kevin Fonseca; Ron Fouchier; Christina Frank; Hans Fredlund; David Freedman; Alexander Friedrich; Ingrid Friesema; Angelika Fruth; Wakaba Fukushima; Luis Furuya-Kanamori; Eric F\u00e8vre; Giovanni Gabutti; Arnaud Gagneur; Wolfgang Gaissmaier; Juan Carlos Galan; Ray Gamble; Sumanth Gandra; Giuliano Garofolo; Petra Gastmeier; Philippe Gautret; Peter Gerner-Smidt; Anne Gershon; Monica Gianfranceschi; Noel Gill; Aharona Glatman-Freedman; Youri Glupczynski; Judith Glynn; Sonia Gomez; Katie Goodman; Magnus Gottfredsson; David Graham; Marc Grandadam; Sophie Granier; Stefan Gravenstein; Christina Greenaway; Gilbert Greub; Emma Griffiths; Thomas Gr\u00f6nthal; Jonathan Gubbay; Nicole Guiso; Jean-Paul Guthmann; Jennifer Guthrie; Bernardo Rafael Guzman Herrador; Stephan G\u00fcnther; Bart Haagmans; Christos Hadjichristodoulou; Sebastian Haller; Anette Hammerum; Mohamed Hammoud; Dag Harmsen; Heli Harvala; Amber Haynes; John P Hays; Jeanne Heil; Marianne Heisz; Bonnie Henry; Niel Hens; Victoria Hernando; Bjorn Herrmann; Roger Hewson; Chelsea Himsworth; Andy Hjortenfeldt; Michele Hlavsa; Heidemarie Holzmann; Yvan Hutin; Benedikt Huttner; V\u00e1clav H\u00f6nig; Judith H\u00fcbschen; Greet Ieven; Derval Igoe; Giuseppe Indolfi; Naoki Inoue; Giuseppe Ippolito; Charlotte Jackson; Michael Jackson; Yves Jackson; Kathryn Jacobsen; Marie Jansson Mork; Meryem Jefferies; Claire Jenkins; Seok Hoon Jeong; Cecilia Jernberg; Dakshika Jeyaratnam; Alan Johnson; Jerker Jonsson; Tsutomu Kageyama; Alexander Kallen; Anu Kantele; Tommi Karki; Basel Karo; Mark Katz; Alyson Kelvin; Yoav Keynan; Hirokazu Kimura; Pete Kinross; Martyn Kirk; Irena Klavs; RM Klevens; Don Klinkenberg; Mirjam Knol; Anke Kohlenberg; Jeroen Kortekaas; Maria Korzeniewska-Kose\u0142a; Rolf Kramer; Tyra Krause; Peter Kreidl; Lothar Kreienbrock; Pavla Krizova; Manuel Krone; Yao Kudjawu; EJ Kuijper; Jaana Kusnetsov; Jeff Kwong; Sharon K\u00fchlmann-Berenzon; Angie Lackenby; Anna Ladogana; Iain Lake; Theresa Lamagni; Sarah Larney; Amparo Larrauri; Heidi Larson; Jeffrey Lazarus; Simon Le Hello; Alexandre Leclercq; Sylvie LeCollinet; Philippe Lehours; Andreia Leite; Katrin Leitmeyer; Isabelle Leparc-Goffart; Tinne Lernout; Daniel Levy-Br\u00fchl; Monica Licker; Diane Lindsay; Theodore Liou; Luiz Lisboa; Nicola Low; Claudia Lucarelli; Catherine Ludden; Christian Lueck; Alexander Lukashev; \u00c5ke Lundkvist; Irja Lutsar; Theodore Lytras; Kristine Macartney; Emily MacDonald; Anna Machowska; Shabir Madhi; Kirsten Maertens; Alexandra Mailles; Helena Maltezou; Georgia Mandilara; Antonella Marangoni; David Marchant; Ulrich Marcus; Giovanni Marini; Robby Markwart; Emily Martin; Helena Martini; Federico Martin\u00f3n-Torres; Josefa Masa-Calles; Alexander Mathis; Peter McIntyre; Jim McMenamin; Paul S. Mead; Adam Meijer; Wim Meijer; Jacques Meis; Hasse Melbye; Tanya Melillo Fenech; Stefano Merler; Jo\u00e3o Mesquita; Peter Michielsen; Sofie Midgley; massimo mirandola; Vivi Miriagou; Thomas Mollet; Dominique Monnet; Ginny Moore; Anne Moorman; Orna Mor; Alain Moren; Pedro l. Moro; Lapo Mughini-Gras; Otilia M\u00e5rdh; K\u00e5re M\u00f8lbak; Marcel M\u00fcller; Anthony Nardone; Umaer Naseer; Pontus Naucler; Nichola Naylor; Ndeindo Ndeikoundam Ngangro; Laura Nellums; Martha Nelson; Nathalie Nicolay; Hubert Niesters; Hiroshi Nishiura; Karsten Noeckler; Harold Noel; Hanna Nohynek; Monika Novak Babic; Sofia Ny; Akihiro Ohkado; George Okoli; Isabel Oliver; Orjan Olsvik; Ryosuke Omori; Nitika Pai; Annalisa Pantosti; Anna Papa; Constantinos Papagiannitsis; John Papp; Dimitrios Paraskevis; Maria Pardos de la Gandara; Sydel Parikh; Maria Paulke-Korinek; Richard Pebody; Tatjana Pekmezovic; Pasi Penttinen; Jose Perez-Molina; Tom Peterman; Efi Petinaki; Martin Petric; Joshua Petrie; Michael Pfaller; Dina Pfeifer; Diamantis Plachouras; Leo Pomar; Stefano Pongolini; Lia Possuelo; Spyridon Pournaras; Edoardo Pozio; Namrata Prasad; Fabian Prasser; Elisabeth Presterl; Jan Prins; Katarina Prosenc; Andrea Pugliese; Joan Puig-Barbera; Mirja Puolakkainen; Mario Ramirez; Jos\u00e9 Tom\u00e1s Ramos Amador; Barbara Rath; Pedro Reis Costa; Vincenzo Restivo; Chantal Reusken; Juliana Reyes Urue\u00f1a; Claire Reynolds; Giovanni Rezza; Flavia Riccardo; Jean-luc Richard; Hans Rieder; Shmuel Rishpon; Caterina Rizzo; Lucy Robertson; Emmanuel Robesyn; Eve Robinson; Susie Roczo-Farkas; Ana Rodrigues; Melissa Rolfes; Magdalena Rosinska; Mirko Rossi; Adam Roth; Markus Ruhnke; Werner Ruppitsch; Daniel Ruzek; Helio Sader; Luisa Salazar; Henrik Salje; Dan Salmon; Vittorio Sambri; Andre Sasse; Gianpaolo Scalia-Tomba; Franciska Schets; Patricia Schlagenhauf; Daniela Schmid; Jonas Schmidt-Chanasit; Isabelle Schuffenecker; Eli Schwartz; Kevin Schwartz; Holly Seale; Harald Seifert; Aaron Seigler; Makeda Semret; Nariman Shahhosseini; Jingjing Shang; Giri Shankar; David Shay; Annette Siedler; Jonne Sikkens; Ian Simms; Benedetto Simone; yannick simonin; Nalini Singh; Vinciane Sizaire; Danuta Skowronski; Rami Sommerstein; Klara Sond\u00e9n; Carla A. Sousa; Ian Spicknall; Gianfranco Spiteri; Marc Steben; Anneke Steens; J Stelling; Andrew Stewardson; Reinhild Strauss; P Strebel; James Stuart; Bertrand Sudre; Carl Suetens; Sheena Sullivan; Lawrence Svenson; Eduardo Taboada; Emi Takashita; Johanna Takkinen; Lara Tavoschi; Nasrin Tayyari Dehbarez; Anne Teirlinck; Laura Temime; Tristan Timbrook; Giota Touloumi; Ramona Trebbien; Athanasios Tsakris; Sarah Tschudin-Sutter; Rainer Ulrich; Anthony Underwood; Magnus Unemo; Tim Uyeki; Marta Valenciano; Palle Valentiner-Branth; Marti Vall-Mayans; Birgit van Benthem; Marita van de Laar; Mark van der Linden; Nicoline van der Maas; Marianne van der Sande; Marieke J. van der Werf; Sonja van Roeden; Ivo van Walle; David Vanduin; Philippe Vanhems; Daisy Vanrompay; Olli Vapalahti; Paula Vasconcelos; Nikos Vasilakis; Alkiviadis Vatopoulos; Jaap Veen; Tomas Vega; Aurelie Velay; Inga Velicko; Akke Vellinga; Harry Vennema; Giulietta Venturi; Lasse Vestergaard; Mafalda Viana; Antonio Vieira; Tatjana Vilibic-Cavlek; Jan Vinj\u00e9; Gianluca Voglino; Alex Vorsters; Georgia Vourli; Tim Wade; Maryse Wanlin; Mary Warrell; Charlotte Warren-Gash; Keith Warriner; Don Weiss; Herbert Weissenboeck; Klaus Weist; Dirk Werber; Guido Werner; Therese Westrell; David Whiley; Heather Whitaker; Ursula Wiedermann; Annelies Wilder-Smith; Hendrik Wilking; Krista Wilkinson; Julie Wilson; Marc Wirden; Carl-Heinz Wirsing von K\u00f6nig; Martin Wolkewitz; Ian Woolley; Zoe Yandle; Lin Yang; Fathiah Zakham; Peter Zarb; Walter Zingg; Milan \u010ci\u017eman."} +{"text": "In the editorial section, Amir Aman Hagos et al. (582) introduce a recent World Health Assembly resolution on emergency care systems.In the news section, Lynn Eaton (585\u2013586) reports on burn-out as an occupational health issue for health care workers. Teguest Guerma talks to Gary Humphreys (587\u2013588) about the early years of the HIV epidemic and her current commitment to training midwives in Ethiopia. Moyeen Uddin et al. (637\u2013641) describe a pilot programme to increase civil registration coverage. Inke Mathauer et al. (620\u2013630) explore options for raising revenue. Blandina Rosalina Bait et al. (597\u2013604) report implementation research from Kupang district.Yuto Maeda et al. (631\u2013636) explain shortfalls in analgesia during childbirth.Sirinad Tiantong et al. (642\u2013644) create a new cooperation strategy.Peter Byass (589\u2013596) examines noncommunicable disease mortality correlates.Daniel O\u2019Keefe et al. (605\u2013611) argue for better indicators of access and impact.Taylor W Burkholder et al. (612\u2013619) propose a rights-based approach."} +{"text": "They also describe severalstrengths and challenges in implementing national-level quality improvement initiativesin low-middle income countries. We concur with the authors that this represents acrucial and important step to improve care and efficiency in resource allocation and tofoster international collaboration in research among these countries.We greatly appreciate the comments from Haniffa et al.Fernando Godinho ZampieriResearch Institute, HCor-Hospital do Cora\u00e7\u00e3o - S\u00e3oPaulo (SP), Brazil.M\u00e1rcio SoaresInstituto D'Or de Pesquisa e Ensino - Rio de Janeiro (RJ), Brazil.Epimed Solutions - Rio de Janeiro (RJ), Brazil."} +{"text": "The Editor-in-Chief has retracted this article (Wang et al."} +{"text": "In the editorial section, Akihiro Nishi et al. (514) discuss what\u2019s needed to reduce Thailand\u2019s annual road traffic death rate to less than 10 deaths per 100 000 people by 2020. Marc Bulterys and Saeed Sadiq Hamid (515) introduce new treatment guidelines for people with chronic hepatitis C virus infection. Sima Barmania (518\u2013519) investigates efforts to improve the health impacts of airports on passengers, staff and people living nearby.Danius Puras talks to Fiona Fleck (520\u2013521) about persuading governments to protect the rights of children with mental disabilities. Kaitlyn Vette et al. (558\u2013567) set thresholds for assessing the severity of pandemic influenza.Nicolas Peyraud et al. (540\u2013547) describe a post-conflict vaccination campaign.Tao Xiong et al. (531\u2013539) study the impact of untreated hypertension in pregnancy. Yi Mu et al. (548\u2013557) track prior caesarean sections and likelihood of vaginal birth.Li Sun et al. (568\u2013577) assess cost\u2013effectiveness of a new screening programme. Emmanuel Andr\u00e9 et al. (522\u2013530) enlist patients to find other people with untreated infections.Caroline Bollars et al. (578\u2013583) describe an adaptation of WHO\u2019s recommended interventions.Jawad and Youssef Fares (584\u2013585) call for better compliance with a cluster munitions treaty.Arian Hatefi et al. (586\u2013588) consider noncommunicable diseases in terms of global susceptibility."} +{"text": "Bulletin. Corrado Barbui et al. (731) describe the lessons learned from a national reform of mental health care provision in Italy. In the editorial section, Michelle McIsaac et al. (730) discuss the findings of a randomized trial on results-based financing, published in this issue of the Sophie Cousins (734\u2013735) reports from Bangladesh on efforts to improve the distribution and capacity of the health workforce. Christian Ntizimira\u00a0talks to Tatum Anderson (736\u2013737) about the challenges of adapting models of palliative care to an African setting.\u00a0Nicolas Meda et al. (750\u2013759) measure the seroprevalence of Hepatitis B and C virus.Adama Diouf et al. (772\u2013781) compare body mass index with deuterium dilution. Daniel Grint et al. (738\u2013749) assess the accuracy of diabetes screening methods in people with tuberculosis.Wu Zeng et al. (760\u2013771) test different ways of implementing results-based financing.Anne Paschke et al. (782\u2013791) describe measures to increase transparency and accountability in national pharmaceutical systems.Amrita Saha & George Alleyne (792\u2013793) argue that noncommunicable diseases should be considered as a global health security issue.Justin J Lang et al. (794\u2013796) argue for measures of cardiorespiratory fitness."} +{"text": "Correction to: Implementation Science (2018) 13:142https://doi.org/10.1186/s13012-018-0823-9Following the publication of this article , the autThe published article contained a discrepancy in Table Table Furthermore, Fig. The correct version of Fig. Mike BradburnFiona McMenemieJason CupittRobert ThompsonNick HarperSimon SleightBelinda CornforthFurthermore, the names of several members of the EPOCH trial group were not processed correctly and should have been processed as follows :The authors also noticed they submitted an incorrect list of authors and the correct list is shown below:EPOCH Trial Group:Abdel Omer; Abhiram Sharma; Abigail Patrick; Adam Paul; Adam Wolverson; Adrian Fawcett; Adrian Jennings; Ajaya Mull; Ajit Sivasankaranand; Alan Morrison; Alastair Ankers; Alastair Rose; Alexandra Scott; Alexandra Williams; Alison Hool; Alison Pickford; Alistair Roy; Alistair Steel; Alister Myers; Almas Rehman; Amanda McCairn; Amanda Stevens; Amir Rafi; Amira Girgis; Amit Shukla; Ana Alegria; Andreas Brodbeck; Andreou Prematie; Andrew Brennan; Andrew Burtenshaw; Andrew Claxton; Andrew Lindner; Andrew Miller; Andrew Thorniley; Andrew White; Andy Thacker; Anil Hermandes; Anita Jhamatt; Anita Sugavanam; Anitha Holtham; Anjay Talwar; Anne Scase; Anthony Parsons; Arnab Bhowmick; Arnth Engel; Ash Prabhudesai; Ashok Raj; Asif Jah; Ayodele Obideyi; Babu Muthuswamy; Bala Maiya; Banwari Agarwal; Barclay Tofte; Belinda Cornforth; Beth Hale; Biju Aravind; Blenk, Karl; Britta O'Carroll-Kuehn; Broad, Dan; Bruce Gibson; Carmen Correia; Carol Mcarthur; Carolyn Way; Catherine Farrow; Catherine Harden; Catherine Jardine; Charles Knowles; Chitre Vivek; Chloe Rochester; Chris Coldwell; Chris Dawson; Chris Homer; Chris Lewis; Chris Nutt; Chris Thorn; Chris Wilson; Christine Bronder; Christopher Macklin; Clare Stapleton; Colin Pow; Craig Lyness; Craig Morris; Dale Vimalchandran; Damian Laba; Dan Freshwater-Turner; Daren Subar; David Bottomley; David Browell; David Gerrard; David Inglis; David Melville; David Monk; David Pogson; David Riddington; David Saunders; David Stanley; Davina Ross-Anderson; Dawn Hales; Dean Millican; Debbie Shaw; Denzil May; Dewi Williams; Dhiraj Ali; Diane Monkhouse; Diane Murray; Dipankar Mukherjee; Dolores Mateo; Dom Hurford; Dominic Sebastian; Donna Doyle; Edward Curtis; Edward Lams; Edyta Niebrzegowska; Elizabeth Hall; Elizabeth Harwood; Emanuel Cirstea; Emma Brennan; Emma Davis; Emma Durant; Emma Leno; Erin Mcilveen; Essam El-Damatty; Esther Cook; Ewen Griffiths; Ewen Harrison; Faisal Baig; Fanus Dreyer; Fenella Welsh; Fiona McMenemie; Flavia Menezes; Flora Bailey; Fran Haigh; Frances Mosley; Francesca Rubulotta; Frankie Dorman; Gabriele Marangoni; Gail Stark; Gail Williams; Gareth Moncaster; Gary Minto; Gavin Bryce; Geoff Watson; Georgia Knight; Gethin Williams; Gillian O'Connell; Giovanni Brescia; Glen Arnold; Gordon Milne; Graham Wilson; Grainne O\u2019Dwyer; Grant Sanders; Greg Lawton; Gudrun Kunst; Guy Finch; Guy Nash; Guy Rousseau; Hamish Noble; Hannah Smith; Harjeet Narula; Hazel Stuart; Heather Pratt; Helen Agostini; Helen Black; Helen Howes; Helen Langton; Helen Porter; Helena Stafford; Hitesh Patel; Huw Davis; Iain Christie; Ian Clement; Ilona Raulusaite; Inga Misane; Ingeborg Welters; Isabella Karat; Jack Carmichael; Jack Parry Jones; Jagdeep Singh; James Bromilow; James Brown; James Burrow; James Harris; James Kirby-Bott; James Limb; Jamie Greenwood; Jane Blazeby; Janindra Warusavitarne; Jason Cupitt; Jay Gokhale; Jay Susarla; Jennifer Edwards; Jennifer Spimpolo; Jenny McLachan; Jenny Ritzema; Jenny Wright; Jens Full; Jeremy Marwick; Jessica Thrush; John Abercrombie; John Corson; John Griffith; John Hopper; John Mackinnon; John Tansley; Jonathan Mccullough; Jonathan Paddle; Jonathon Barker; Jonathon Mole; Jonny Wilkinson; Josef Watfah; Jost Mullenheim; Julian Sonksen; Julian Stone; Julie Colley; Julie Furneval; Julie Wakeford; Julie Wollaston; Justin Woods; Jyrki Karvonen; Kaighan Lynne; Kamal\u00a0Aryal; Kar\u200bthik\u200b Surendran; Karan\u200b Verma; Karen Burns; Karen Simeson; Karvonen Jyrki; Kate Wong; Kathryn Cain; Kathryn Gill; Katie Cooke; Keiarash Jovestani; Kenneth Adegoke; Kevin Rooney; Kevin Sim; Khaled Razouk; Kim Jemmet; Kirosh\u00a0\u200bShankar; Kirsty Baillie; Kirsty Everingham; Krishnamurthy Somasekar; Kumar Panikkar; Lampros Liasis; Laura Graham; Laura Rooney; Lawrence Wilson; Lee Baldwin; Leilani Cabreros; Liam\u200b Hudson; Linda Graham; Lindsay Bailey; Lorna Burrows; Louise Bell; Lynn Stewart; Lynn Taylor; Lynsey Kightly; Magdy Khater; Maitra Ishaan; Majed Al Shama; Makvana Sonia; Malcolm Sim; Malcolm Watters; Manab Mohanty; Mansoor Akhtar; Mansoor Sange; Marcus Wood; Maria Bews-Hair; Maria Lawson; Marion Obichere; Mark Blunt; Mark Cartmell; Mark Coleman; Mark Henwood; Mark Munro; Mark Pulletz; Mark Snazelle; Mark Watson; Mark Wilkinson; Marta Campbell; Marta Januszewska; Martin Leuwer; Martin Northey; Martin Stotz; Martyn Cain; Massimo Varcada; Matt Gardner; Matt Outram; Matthew Gaughan; Matthew Tutton; Maurizio Cecconi; Meghna Sharma; Melanie Tan; Michael Chadwick; Michael Crabtree; Michael Gillies; Michael Karlikowski; Michael Machesney; Michael Martin; Mike Bradburn; Mike Gay; Nabil El-Masry; Nabua Gerstina; Nada Hadi; Nandita Divekar; Nat Natarajan; Natalie Dickinson; Nathan Pushpa; Nathan Borgeaud; Nazar Abdul; Nazzia Mirza; Neil Cruickshank; Neil Flint; Neil Kukreja; Nicholas Watson; Nick Bunker; Nick Harper; Nick Mason; Nicola Cook; Nicola Lythell; Nicola Radford; Nicola Stanix; Nicole Robin; Nigel Hollister; Nigel Suggett; Niko Van De Velde; Nikolaos Makris; Olga Tucker; Oliver Hill; Oliver Zuzan; Oluremi Odejinmi; Otto Mohr; Paddy Collins; Panna Patel; Paul Harrison; Paul Mclaren; Paul O'Loughlin; Paul Ziprin; Paul Noble; Pedro Cunha; Peeyush Kumar; Peter Alexander; Peter Chan; Peter Davies; Peter Fitzgerald; Peter Lamb; Peter Richardson; Phil Dodd; Phil Hopkins; Phillippa Pemberton; Phoebe Bodger; Pieter Bothma; Piroska Toth-Tarsoly; Preeti Kuduvalli; Qamar Iqbal; Rachael Craven; Rai Kuldip; Raj Patel; Rajesh Dumpala; Raman Guruswamy; Ramani Moonesinghe; Rame Sunthareswaran; Ramesh Rajagopal; Ranjit Ganepola; Raoul Benlloch; Razeen Mahroof; Rich Gibbs; Richard Hartopp; Richard Haslop; Richard Howard-Griffin; Richard Morgan; Richard Pugh; Richard Wharton; Ricky Lewis; Rob Chambers; Robert DeBrunner; Robert Shawcross; Roger Townsend; Roshan\u00a0Lal; Rovan D'souza; Rowena Felipe; Roy Fernandes; Ruth Griffin; Ruth Thomas; Sally Roth; Sam Andrews; Sam Waddy; Samer Doughan; Sami Farhat; Sami Ijaz; Sandeep Varma; Sanjay Wijeykoon; Sara Pick; Sarada Gurung; Sarah Beavis; Sarah Bowery; Sarah Buckley; Sarah\u00a0Downey; Saravanna Sagadai; Satish Singh; Savvas Papagrigoriadis; Sean Cope; Sean McAfee; Sean Mcmullan; Senthil Nadarajavan; Sergei Vaganov; Shameem Sarfi; Sheshagiri Bengeri; Shrisha Shenoy; Shubha Vashisht; Sian Bhardwaj; Sid Riddington; Simon Bailey; Simon Fletcher; Simon Gibson; Simon Harris; Simon Hester; Simon Parrington; Simon Sleight; Simon Smart; Singh Gursevak; Somi Desikan; Sophie Noblett; Stacy Hodges; Stas Janokowski; Stefan Pulsa; Stelios Chatzimichail; Stella Vig; Stephanie Bell; Stephen Baxter; Stephen Harris; Stephen Lake; Steve Fletcher; Steve Hutchinson; Steven Henderson; Stewart Prestwich; Stuart Mercer; Sudha Garg; Surjait Singh; Susan Dowling; Susan Jain; Susan Moug; Susan Tyson; Susie Baker; Syed Iftikhar; Tabitha Tanqueray; Taj Saran; Tamas Szakmany; Tamsin Rope; Tamzin Cuming; Tanuja Shah; Tariq Hussein; Tezas Stergios; Therese Murray; Thomas Evans; Thomas Medici; Thomas Parker; Tim Campbell-Smith; Tim Geary; Tim Harvard; Tim White; Tom Abbott; Tom Edwards; Tom Morgan-Jones; Tom Owen; Tomas Jovaisa; Una McNelis; Valerie Hilton; Vamsi\u00a0Velchuru; Vanessa Linnett; Vanessa Tucker; Veena Naik; Victoria Banks; Vishal Patil; Vivek Chitre; Vlad Kushakovsky; Wael Khalaf; Wayne Wrathall; Will Brady; Xavier Escofet; Yasser Mohsen; Ying Hu.The original article has been corrected. The publisher apologizes to the readers and the authors for any inconvenience caused by these errors."} +{"text": "In the editorial section, Beverley M Essue et al. (442) discuss policies designed to address the high cost of care for chronic kidney disease in India. Yvan Hutin et al. (443) describe milestones in the reduction of hepatitis B infections. Sophie Cousins (446\u2013447) reports on cholera vaccination programmes in crisis settings. Yoshitake Yokokura tells Fiona Fleck (448\u2013449) how Japan proposes to direct primary health care services towards the needs of an ageing population. Marina Daniele et al. (450\u2013461) study the inclusion of male partners in maternity care.Catherine Barker Cantelmo et al. (462\u2013470) estimate the costs of a 5-year national health plan.Mark Goodchild & Rong Zheng (506\u2013512) assess the impact of a recent tax increase.Aaron Richterman et al. (471\u2013483) collect the evidence of effects on clinical outcomes.Kate Whitford et al. (484\u2013497) review long-term impacts of infant immunization programmes. Jennifer Cooper et al. (498\u2013505) propose a classification for patient-safety incidents."} +{"text": "This month\u2019s theme is universal health coverage, linked to the Prince Mahidol Award Conference in Bangkok, Thailand.In the editorial section, Viroj Tangcharoensathein et al. (78) explain why it is time to deliver on political promises of extending health coverage to all people. Elizabeth Mason et al. (79) focus on the particular needs of women, children and adolescents. Susan P Sparkes and Joseph Kutzin (80) provide a reminder of the need to include HIV prevention and care services. In the news section, Gary Humphreys (83\u201384) reports on efforts to coordinate coverage over the geographical expanse of the Caribbean islands. Midori de Habich speaks to Gary Humphreys (85\u201386) about policies leading to universal health coverage in Peru.Giorgio Cometto et al. (109\u2013116) discuss good practices in health workforce management.Charles B Holmes et al. (87\u201394) weigh the pros and cons of targeted programmes.Inke Mathauer et al. (132\u2013139) provide a global overview of pooling reforms.Sarah Louise Barber et al. (95\u201399) recommend costing exercises for national plans to increase coverage.Ileana Vilcu et al. (126\u2013131) link health information systems to strategic purchasing reforms.Bocar Mamadou Daff et al. (100\u2013108) examine national financial protection measures.Kanang Kantamaturapoj et al. (117\u2013125) review participatory approaches used to inform health financing policies.Viroj Tangcharoensathein et al. (140\u2013145) dissect the political economy of an universal coverage scheme.Mickey Chopra et al. (146\u2013148) emphasize the need to achieve equitable immunization coverage."} +{"text": "This month\u2019s special theme is hearing loss: building capacity for a public health response. In editorials, Adrian C Davis & Howard J Hoffman (646) call attention to an increased global prevalence and incidence of hearing loss. Shelly Chadha et al. (647) examine health system requirements for hearing care services.Gary Humphreys (650\u2013651) reports on efforts to reduce noise exposure in the workplace. Bolajoko Olusanya talks to Sophie Cousins (652\u2013653) about her experience with a late diagnosis of hearing impairment, and her efforts to improve the hearing care provided to children in Nigeria.Michael Yong et al. (699\u2013710) review policies to improve the availability of adults\u2019 hearing aids.Jonathan J Suen et al. (681\u2013690) describe community provision of hearing care.Wakisa Mulwafu et al. (654\u2013662) find substantial loss to follow-up.Peter R Thorne et al. (719\u2013721) map the provision of hearing care services.Susan Eksteen et al. (672\u2013680) use mobile phones to screen preschool children.Pittayapon Pitathawatchai et al. (663\u2013671) report on a pilot programme in four hospitals.Mahmood F Bhutta et al. (691\u2013698) estimate needs for primary care providers, audiologists, ear, nose and throat specialists, speech therapists and teachers. Joseph J Murray et al. (711\u2013716) explore the contribution of signed languages.Karissa L LeClair and James E Saunders (722\u2013724) argue for an holistic approach. De Wet Swanepoel et al. (717\u2013718) uncover strengths and weaknesses of mobile applications.Bolajoko O Olusanya et al. (725\u2013728) request a review of terminology."} +{"text": "Ciber de Enfermedades Respiratorias - CibeRes is an initiative of Instituto de Salud Carlos \u2162."} +{"text": "In the editorial section, M Claire Greene et al. (246) note a lack of attention to substance use disorders in refugee populations. Anayda Portela et al. (247) ask guideline developers to account for the complexity of health systems.Andrey Shukshin (250\u2013251) reports on Kazakhstan\u2019s efforts to reform its health system four decades after the Declaration of Alma Ata. Mirfin Mpundu tells Gary Humphreys (252\u2013253) about the challenges faced by African countries looking to contain antimicrobial resistance. Evelina Martelli et al. (259\u2013269) evaluate the impact of a programme designed to increase birth registration.Gustavo Corr\u00eaa et al. (306\u2013308) argue for broader use of immunization data.Suman Chakrabarti et al. (270\u2013282) analyse equity and coverage of child services.Bhargava Mullapudi et al. (254\u2013258) estimate gaps in access to pediatric surgery.C\u00e9cile Aenishaenslin et al. (283\u2013289) point out that better evidence is needed. Keiko Osaki and Hirotsugu Aiga (296\u2013305) argue for the adaption of home-based records.Robin Ireland et al. (290\u2013296) call for scrutiny of sponsorship and advertising."} +{"text": "With interest we read the recent paper by Charbonneau et al. tackling side effects of angiotensin-converting enzyme inhibitor (ACEI) therapy during hypovolemic shock in mice by using icatibant, a specific bradykinin beta 2 receptor antagonist . They de"} +{"text": "In the editorial section, Amina J Mohammed and Tedros Adhanom Ghebreyesus (590) map the links between human health and sustainable development. Shambhu Acharya et al. (591) review the research published on this theme.Shelly Chadha et al. (592) call for papers on the topic of hearing loss. In the news section, G\u00f6ran Tomson tells Fiona Fleck (595\u2013596) how scientists can help governments make economic progress without damaging the environment or depleting natural resources. Sima Barmania (597\u2013598) reports on attempts to design sustainable airports and to mitigate harms to users. Jonathan Cylus et al. (599\u2013609) compare methods for measuring catastrophic health expenditure. Hui Wang et al. (610\u2013620) analyse data on household health expenditures. Kira Fortune et al. (621\u2013626) describe health promotion initiatives.Judith Bueno de Mesquita et al. (627\u2013633) support the use of human right accountability reviews to monitor progress.Timothy Ken Mackey et al. (634\u2013643) propose a framework for reducing corruption in the health sector. Mary Manandhar et al. (644\u2013653) critique a view of gender that is limited to women.Ahmad Reza Hosseinpoor et al. (654\u2013659) discuss the quantification of health inequalities. Asiya Odugleh-Kolev & John Parrish-Sprowl (660\u2013661) explain why community engagement is needed for universal health coverage.Katie Clifford et al. (662\u2013664) examine the implications of poor quality veterinary medicines."} +{"text": "In the editorial section, Linda-Gail Bekker et al. (170) examine governance of the HIV response in comparison to other global health programmes. Woranan Witthayapipopsakul et al. (171) announce an upcoming theme issue and call for papers on universal health coverage.In the news section, Andrey Shukshin (174\u2013175) reports on greater availability of cochlear implants in the Russian Federation. Paula Radcliffe explains to Gary Humphreys (176\u2013177) why long-distance runners are particularly concerned about air pollution and introduces a clean air initiative of the International Associations of Athletics Federation.Nigar Nargis et al. (221\u2013229) find that tobacco taxes are insufficient to reduce consumption. Jue Liu et al. (230\u2013238) describe progress in hepatitis B control.Meru Sheel et al. (178\u2013189) evaluate an early warning, alert and response system.Harriet Jones et al. (200\u2013212) study policy implementation.Suladda Pongutta et al. (213\u2013220) convey lessons learned from a health promotion foundation.Peishan Ning (190\u2013199) track changes in reporting methods.Seye Abimbola et al. (239\u2013241) compare HIV to noncommunicable diseases.Damien Ming et al. (242\u2013244) propose harvesting data from connected diagnostic platforms."} +{"text": "In the editorial section, Ivo Kocur et al. (666) introduce this theme issue on the future of eye care in a changing world.Karin Weyer et al. (667) discuss the improvements needed in treatment regimens for drug-resistant tuberculosis.In the news section, Jack Latimore (670\u2013671) reports on efforts to address eye-care needs of Australia\u2019s Indigenous people. In an interview, Haroon Awan explains to Fiona Fleck (672\u2013673) how several low- and middle-income countries are incorporating eye care into their national health strategies.Anthea M Burnett et al. (682\u2013694) gather the evidence on school-based eye-care services.Venkata SM Gudlavalleti et al. (705\u2013715) describe how avoidable blindness is handled.Omar Salamanca et al. (674\u2013681) implement a diabetic retinopathy referral network.Kristof Wing et al. (716\u2013722) describe the creation of a national eye-care service.Jacqueline Ramke et al. (695\u2013704) review the evidence for national universal eye health plans.Sarity Dodson et al. (723\u2013725) compare frameworks for interventions needed to eliminate trachoma in 57 countries.Islay Mactaggart et al. (726\u2013728) outline methodological improvements to the rapid assessment of avoidable blindness."} +{"text": "The editorial team is grateful to all the experts who dedicated their time to peer-review our articles over the past 12 months. We wish to thank them and all those whose names are not listed below but gave helpful advice and support. We apologise to any reviewers whose names we may have missed.Our reviewers in 2018 were:Preben Aavitsland; Winston Abara; Mark Achtman; Blythe Adamson; Karam Adel Ali; Amos Adler; Cornelia Adlhoch; Tobias Alfven; Franz Allerberger; Matthias an der Heiden; Nick Andrews; Augustina Annan; Alberto Antonelli; Julien Arino; Laurence Armand; Olle Aspevall; Agoritsa Baka; Biswajit Banik; Lucie Bardet; Ian Barr; Joel Barratt; Mette Bartels; Luisa Barzon; Elizabeth Beech; Michael Behnke; Romeo Bellini; Edward Belongia; Beatrice Bercot; Anne Berger-Carbonne; Dag Berild; Kathy Bernard; Xavier Bertrand; Marianne Besnard; John Besser; Janneke Bil; Zeno Bisoffi; Kathe E Bjork; Dominique Blanc; Aur\u00e9lie Bocquier; Pierre-Yves Boelle; Shelly Bolotin; Magnus Boman; Paolo Bonanni; Robert A. Bonomo; Marc Bonten; Michael Borg; Maria Borrego; Arnold Bosman; Elisabeth Botelho-Nevers; Julie Bottero; Charles Boucher; Nathalie Boulanger; Herve Bourhy; Naomi Boxall; Jean-Philippe Brandel; Elizabeth Brickley; Tom Britton; Eeva Broberg; Stef Bronzwaer; Tal Brosh-Nissimov; Florence Brossier; Kevin Brown; Mia Brytting; Udo Buchholz; Ann Burchell; Zahid Butt; Valentina Cambiano; Rosa Cano; Gioia Capelli; Saulius Caplinskas; Alessandra Carattoli; Yehuda Carmeli; Jo\u00e3o Andr\u00e9 Carri\u00e7o; Pamela Cassiday; Nadim Cassir; Alejandra Castanon; Vicki Chalker; Meera Chand; Herve Chaudet; Shahnaz Akhtar Chaudhry; Sheng Chen; Gerardo Chowell; Dimple Chudasama; Andrew Clark; Jan Clement; Michelle Cole; Simon Collin; Victor Corman; Helena Cortes Martins; Suzanne Cotter; Simon Cottrell; Suzie Coughlan; Natacha Couto; Susan Cowan; Benjamin Cowling; Anne Claude Cremieux; Natasha Crowcroft; Fortunato D'Ancona; Viktor Dahl; Tim Dallman; Eric Dannaoui; Birgitta de Jong; Gaston De Serres; Henry de Vries; Katja de With; Aleksander Deptula; Tarik Derrough; Jean-Claude Desenclos; Ram Dessau; Joanne Dillon; Roger Dodd; Dragoslav Domanovic; Lisa Domegan; Christina Domingo; Michel Drancourt; Mike Drebot; Erika Duffell; Tim Eckmanns; Paul Effler; Carina Eklund; Sascha Ellington; Delia Enria; Onder Ergonul; Nagy Erzs\u00e9bet; Concepcion Estivariz; Martin Exner; David Eyre; Anna-Bella Failloux; Christopher Fairley; Gerhard Falkenhorst; Heinz Feldmann; Peter Feng; Olivier Ferraris; Marc Fischer; Julia Fitzner; Brendan Flannery; Stefan Flasche; Agnes Fleury; Ana Gloria Fonseca; Ivo Foppa; Pieter Fraaij; Christina Frank; Eelco Franz; Scott Fridkin; Hilbert Friederike; Alexander Friedrich; Ingrid Friesema; Niels Frimodt-M\u00f6ller; Angelika Fruth; Giovanni Gabutti; Pierre Gallian; Giuliano Garofolo; Philippe Gautret; Julianne Gee; Peter Gerner-Smidt; Michael Geschwind; Tommaso Giani; Christopher Gilpin; Christophe Ginevra; Aharona Glatman-Freedman; Anja Globig; Y Glupczynski; Pere Godoy; Daniel Golparian; Douglas Goodin; Ewelina Gowin; Luigi Gradoni; Christina Greenaway; Lutz Guertler; Barbara Gunsenheimer-Bartmeyer; Bernardo Rafael Guzman Herrador; Huldrych G\u00fcnthard; Bart Haagmans; Katrine Bach Habersaat; Marisa Haenni; Susan Hahne; \u00c1gnes Hajdu; Sebastian Haller; Anette Hammerum; H\u00e5kan Hanberger; Stephan Harbarth; Pia Hardelid; Sally Hargreaves; Heli Harvala; Jaythoon Hassan; Barbara Hauer; Joana Haussig; Peter Helbling; Rene Hendriksen; Niel Hens; Bjorn Herrmann; Sereina Herzog; Kristen L Hess; Ursel Heudorf; Ole Heuer; Roger Hewson; Lauri Hicks; Andy IM Hoepelmann; Sven Hoffner; Michael Hogardt; Kristy Horan; Te-Din Huang; Yvan Hutin; Michael H\u00f6hle; Giovanni Ianiro; Derval Igoe; Giuseppe Ippolito; Jennifer Isenor; Michael Jackson; Lynn Janssen; Claire Jenkins; Seok Hoon Jeong; Samo Jeverica; Silvia Jimenez-Jorge; Kari Johansen; Alan Johnson; Helen Johnson; Pikka Jokelainen; Michael Jordan; Loic Josseran; Achim Kaasch; Sarah Kabbani; Annemarie Kaesbohrer; Bernard Kaic; Ilias Karaiskos; Tommi Karki; Peter Keller; Volkhard Kempf; Yury Khudyakov; Mirjam Knol; Anke Kohlenberg; Axel Kola; Gerard Krause; Peter Krause; Peter Kreidl; Sanjeev Krishna; Paul Krogstad; Anna Kuehne; Katrin Kuhn; Ed Kuijper; Heinke Kunst; Robin K\u00f6ck; Angie Lackenby; Shamez Ladhani; John Lambert; Amparo Larrauri; Heidi Larson; Simon Le Hello; Carol Leitch; Tinne Lernout; Joseph Lewnard; Bruno Lina; Andrew Lloyd; Shawn Lockhart; Nicholas Loman; Pierluigi Lopalco; Jens Lundgren; Yaniv Lustig; Michael S. Lyons; Outi Lyytik\u00e4inen; Daniel L\u00e9vy-Bruhl; Noni Mac Donald; Kristine Macartney; Liane Macdonald; Colin Mackenzie; Shelley S. Magill; Anna-Pelagia Magiorakos; Barbara Mahon; Helena Maltezou; Jean-Michel Mansuy; Ulrich Marcus; Lauri E. Markowitz; Nicolee Martin; Clara Mar\u00edn; Maria Teresa Mascellino; Virginie Masserey; Sarah Mbaeyi; Scott McDonald; Jeffrey McFarland; Peter McIntyre; Huong McLean; Jim McMenamin; Nicholas Medland; Conor Meehan; Adam Meijer; Elissa Meites; Angeliki Melidou; Kassiani Mellou; Ella Mendelson; Jeffrey W. Mercante; Mathias Merker; Stefano Merler; Jaimie Meyer; Claire Midgley; Sofie Midgley; Andrei Mihalca; Massimo Mirandola; Hamish Mohammed; Orna Mor; Dearbhaile Morris; Anne Mosnier; Jo\u00ebl Mossong; Judith Mueller; Sanjin Musa; Didier Musso; Luise M\u00fcller; Rajeshwari Nair; Dilip Nathwani; Warrick Nelson; Lina Nerlander; Emanuele Nicastri; Matthias Niedrig; Harold Noel; Paulo Nogueira; Torbj\u00f6rn Nor\u00e9n; Hans-Dieter Nothdurft; Norbert Nowotny; Baltazar Nunes; Claudia Nunes Duarte dos Santos; Mari Nyg\u00e5rd; Colm A. O'Morain; Sophie Octavia; Nicholas Ogden; Isabel Oliver; Jes\u00fas Oteo; W John Paget; Ana M. Palomar; Constantinos Papagiannitsis; Dimitrios Paraskevis; Cyra Patel; John Paul; Lucia Pawloski; Lara Payne; Malik Peiris; Camille Pelat; Pasi Penttinen; Freddy Perez; Jeannine Petersen; Lyle Petersen; Martin Petric; Juraj Petrik; Yvonne Pfeifer; Anastasia Pharris; Nick Phin; Mathieu Picardeau; Roan Pijnacker; Tamara Pilishvili; Alessandro Pini; Jillian Pintye; Diamantis Plachouras; Laurent Poirel; Jeanine Pommier; Garyfallia Poulakou; Jan Prins; Katarina Prosenc; Andrea Pugliese; Celine Pulcini; David W Purcell; Helen Quinn; Sophie Quoilin; Veronique Raimond; Mario Ramirez; Brian Raphael; Vincenza Regine; Renata Reis; Chantal Reusken; Sandra Reuter; Giovanni Rezza; Anders Rhod Larsen; Dale Rhoda; Caterina Rizzo; Emmanuel Robesyn; Eve Robinson; Jes\u00fas Rodr\u00edguez-Ba\u00f1o; T Rolling; Laurence Roope; Roberto Rosa; John Rossen; Mirko Rossi; Gian Maria Rossolini; Adam Roth; Maja Rupnik; Werner Ruppitsch; Claudia Ruscher; Kristi R\u00fc\u00fctel; Joaquin Salas-Coronas; David Samara; Jussi Sane; Sabine Santibanez; Loredana Sarmati; Andre Sasse; Silvia Sauleda Oliveras; Gianpaolo Scalia-Tomba; Gaia Scavia; Robert Schlaberg; Patricia Schlagenhauf; Jonas Schmidt-Chanasit; Eli Schwartz; Kevin Schwartz; Torsten Semmler; William Shafer; Ian Simms; Benedetto Simone; Martin Singer; Vitali Sintchenko; Teemu Smura; Heidi Soeters; Hanna Soini; Carla A. Sousa; Christina Spiropoulou; Gianfranco Spiteri; Russell Stafford; Helen Stagg; Gerold Stanek; Pawel Stefanoff; Claudia Stein; Chen Stein-Zamir; Rune Stensvold; Maja Subelj; Carl Suetens; Kathryn Suh; Jonathan Suk; Sheena Sullivan; Arnfinn Sundsfjord; Johanna Takkinen; Arnaud Tarantola; Anders Tegnell; Noemia Teixeira de Siqueira-Filha; Laura Temime; Nicola Thompson; Claire Thorne; Salla Toikkanen; M Tongo; Mia Torpdahl; Soren Uldum; Tim Uyeki; Palle Valentiner-Branth; Snigdha Vallabhaneni; Alex van Belkum; Wim Van Bortel; Marita van de Laar; Kees. C. van den Wijngaard; Arie van der Ende; Maarten F. Schim Van Der Loeff; Marianne van der Sande; Marieke J. van der Werf; Gary Van Domselaar; Sofie van Dorp; Jakko Van Ingen; Olli Vapalahti; Carmen Varela Martinez; Alkiviadis Vatopoulos; Alida Veloo; Harry Vennema; Salvador Vergara-L\u00f3pez; Pierre Verger; Lasse Vestergaard; Jan Vinj\u00e9; Leo Visser; Margreet Vos; Elena Vovc; Julio A. V\u00e1zquez Moreno; Timothy Walsh; Jan Walter; Doreen Walther; Jiangrong Wang; Francois Xavier Weill; Hillard Weinstock; Dirk Werber; Guido Werner; Henrik Westh; Therese Westrell; David Whiley; Ole Wichmann; Andreas Widmer; Annelies Wilder-Smith; Hendrik Wilking; Christopher Williams; Carl-Heinz Wirsing von K\u00f6nig; Edwina Wright; Lin Yang; Abdool Yasseen; Hans L Zaaijer; Katherina Zakikhany; Philipp Zanger; Gernot Zarfel; Herve Zeller; Walter Zingg; Ines Zollner-Schwetz; Huachun Zou; Phillip Zucs."} diff --git a/PMC_clustering_97.jsonl b/PMC_clustering_97.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..9145bbb629ea77829a4b3a29d58d54ef557e9132 --- /dev/null +++ b/PMC_clustering_97.jsonl @@ -0,0 +1,970 @@ +{"text": "In the original article, there was a mistake in"} +{"text": "Following publication of the original article , the aut"} +{"text": "Following publication of the original article , the aut"} +{"text": "Following the publication of the original article , the aut"} +{"text": "In the original version of our article , insuffi"} +{"text": "Following publication of the original article , the aut"} +{"text": "Following publication of the original article , the aut"} +{"text": "Following publication of the original article , the aut"} +{"text": "Following publication of the original article , the aut"} +{"text": "After online publication of the article , the fir"} +{"text": "In the original article , there w"} +{"text": "Following publication of the original article , the aut"} +{"text": "Following publication of the original article , the aut"} +{"text": "After careful consideration, we found that"} +{"text": "Following publication of the original article , an erro"} +{"text": "Unfortunately, the original version of the article containe"} +{"text": "In the original article, we missed the parallel work by"} +{"text": "Recently, we have found a mistake in Figure"} +{"text": "In Additionally, Eqs"} +{"text": "After publication of the original article , the aut"} +{"text": "Following publication of the original article , the aut"} +{"text": "Following the publication of this article , it has"} +{"text": "Vugia, was incorrect. 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original publication of this article , the cor"} +{"text": "Following publication of the original article , we were"} +{"text": "Following publication of the original article , the aut"} +{"text": "Following publication of the original article , the aut"} +{"text": "In the original publication , there w"} +{"text": "In the original publication , Figure"} +{"text": "In the original publication , there w"} +{"text": "In the original publication , there w"} +{"text": "Consequently, the corrected version of"} +{"text": "In the original publication , there w"} +{"text": "In the original publication , there w"} +{"text": "In the original version of thisarticle, an error was identified in"} +{"text": "In the original publication , there w"} +{"text": "In the original publication , there w"} +{"text": "In the published article, there was an error in"} +{"text": "In the original publication , the ori"} +{"text": "In the original publication , there w"} +{"text": "In the original publication , 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study , we disc"} +{"text": "After publication of this manuscript , we note"} +{"text": "After publication of this article , the aut"} +{"text": "After publication of this work , we note"} +{"text": "Following the publication of this article , the aut"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of this work , we note"} +{"text": "Unfortunately, the original version of this article containeAnna-Kristin Brettschneider"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of this work , it was"} +{"text": "After the publication of the original article , it was"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "During production of the original article , an over"} +{"text": "After publication of the original article , the pub"} +{"text": "Unfortunately, the original version of this article contains"} +{"text": "Unfortunately, the original 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containe"} +{"text": "Due to a publisher error, this article was orig"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, after publication of this article it was n"} +{"text": "After publication of this study , the aut"} +{"text": "Upon publication, a mistake in Additional file"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, after publication of this article , it was"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After the publication of this article , it was"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, after publication of this article it was n"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of the original article , it came"} +{"text": "Unfortunately, the 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in"} +{"text": "ErratumAfter the publication of this work , we noti"} +{"text": "Unfortunately, the original version of this article containeRalf Moreno Garcia"} +{"text": "After publication of this abstract , we foun"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, during production of this article , an erro"} +{"text": "After publication of the original article , the aut"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, after publication of this article it was n"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Due to a publisher error, the original version of this article was regr"} +{"text": "Following publication of our article , we founThe corrected"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of our recent Poster presentation , we noti"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of the original article , it was Additionally, Additional file"} +{"text": "After publication of the original article , it was"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of this work , we note"} +{"text": "During production of the original article , an erro"} +{"text": "Upon publication of this article , it was"} +{"text": "Unfortunately, after publication of the article it was n"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original 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we noti"} +{"text": "After publication of this study , we noti"} +{"text": "Following the publication of the original article , an erro"} +{"text": "After publication of the original article , it came"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article did not"} +{"text": "Following publication of our article , it was"} +{"text": "Unfortunately, after publication of this article , it was"} +{"text": "In the originally published version of this article , incorre"} +{"text": "Following publication of our article , we beca"} +{"text": "Upon publication of this article , it was"} +{"text": "After publication of the original article , the aut"} +{"text": "In the original article , there w"} +{"text": "After publication of this work , we note"} +{"text": "ErratumAfter the publication of this work , we noti"} +{"text": "After publication of the original article , it came"} +{"text": "After publication of this work , we noti"} +{"text": "Upon publication, it was noticed that the acknowledgements section of this article should a"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "During the production process for this article , Theorem"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of this work , we note"} +{"text": "After publication of our article , it came"} +{"text": "Unfortunately, the original version of this article (Zhang et al."} +{"text": "After publication of this work , it was"} +{"text": "After publication of this study , we noti"} +{"text": "After publication of the original article , it came"} +{"text": "After publication of the original article , it was"} +{"text": "While compiling the article one of t"} +{"text": "Unfortunately, after publication of this article , it was"} +{"text": "Unfortunately, after publication of the original version of this article , we noti"} +{"text": 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article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of this work , we note"} +{"text": "ErratumUnfortunately, the original version of this article containe"} +{"text": "For the previous publication of our article , Figure"} +{"text": "Unfortunately, after publication of this article , an erro"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of this article , the aut"} +{"text": "After the publication of this work , we noti"} +{"text": "Unfortunately, the original version of this article (Gupta & Soto"} +{"text": "After publication of this study , the aut"} +{"text": "Following publication of our article , it has Precision = Burden ="} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Please view the complete, correct equation here:"} +{"text": "After the 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containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of this study , the aut"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "The legends for Please see the complete, corrected"} +{"text": "After the publication of this work , the aut"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of this work , we disc"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of the original article , the aut"} +{"text": "After the publication of this work , we noti"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publishing of the paper , substan"} +{"text": "After publication of the original article , an erro"} +{"text": "Following publication of this work , the aut"} +{"text": "In the original publication, error was found in Fig"} +{"text": "After publication of this article , the autThe original article has been"} +{"text": "The corrected In the original publication , there w"} +{"text": "In the original publication , there w"} +{"text": "Following publication of the original article , the aut"} +{"text": "Following publication of the original article , the aut"} +{"text": "In the original publication , there w"} +{"text": "Following the publication of the original article , the aut"} +{"text": "After publication of this article , the autThe original article has been"} +{"text": "In the original publication , errors"} +{"text": "After publication of this article , the autThe original article has been"} +{"text": "Following publication of the original article , it was"} +{"text": "In the original publication , there 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of this article, some data in Fig."} +{"text": "Due to a production error, in the original article, references \u201315 were"} +{"text": "In this article , the aff"} +{"text": "In this article , there w Table"} +{"text": "There are glycopeptide structures missing in both Also, in"} +{"text": "In the original HTML version of this article , there w"} +{"text": "In the original article, there was a mistake in"} +{"text": "Following publication of the original article , it was"} +{"text": "In this article , there w"} +{"text": "Following the publication of the original article , we were"} +{"text": "Following publication of the original article , an erro"} +{"text": "Following publication of the original article , it was The original article has been"} +{"text": "Following publication of the original article , the aut"} +{"text": "Figs In For For Please see the complete, correct Figs"} +{"text": "Following publication of the original article , the aut"} +{"text": "In the original 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+{"text": "Following publication of the original article , it came"} +{"text": "We thank Dr. Lawler for his letter , which a"} +{"text": "After publication of this article , it was"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "After publication of the article , it has"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "In the publication of this article , there w"} +{"text": "Following publication of the original article , the aut"} +{"text": "After publication of this article , it was"} +{"text": "Unfortunately, the original version of this article containe"} +{"text": "Upon publication of the original article , it was"} +{"text": "After publication of the article , it has"} +{"text": "After publication of the article , it has"} +{"text": "After publication of the article , it has"} +{"text": "After publication of the article , it was"} +{"text": "The correct sentence is: Here, a Micropaque"} +{"text": 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issue,"} +{"text": "Following the publication of this article , it was"} +{"text": "In our article , we citeOur discussion of"} +{"text": "Sir,The finding reported by"} +{"text": "Sir,In the March 8 issue,"} +{"text": "Sir,We read with great interest the article by"} +{"text": "During the publication process of this work , we note"} +{"text": "After the publication of this work , we beca"} +{"text": "Sir,The paper by"} +{"text": "Sir,I am delighted to note that Prof. Dalal has brou"} +{"text": "After publication of this work , we note"} +{"text": "Dear Editor,We thank Sharma for the"} +{"text": "After the publication of this work , we beca"} +{"text": "Sir,We are thankful for this comment"} +{"text": "Dear Editor,I thank the reader for appr"} +{"text": "After publication of this work , we note"} +{"text": "Since publication of our article , we have"} +{"text": "Potency and toxicity data showed that, generally, the Ph"} +{"text": "After the publication of this research article , the aut"} +{"text": "After the publication of this work , we disc"} +{"text": "Sir,The recent publications of On a further point,"} +{"text": "Unfortunately, after publication of this article , it was"} +{"text": "After publication of the original article , an erro"} +{"text": "Due to an error during production, the column title of"} +{"text": "Following publication of the original article , one of"} +{"text": "After publication of the original article , the aut"} +{"text": "Dear Editor,We appreciate the interest in our o"} +{"text": "Following publication of the original article , the aut"} +{"text": "After publication of the original article , the aut"} +{"text": "Due to an error during production, the legend presented in"} +{"text": "In the original publication of this article , the spe"} +{"text": "In the original article, there was a mistake in"} +{"text": "In the original article, there were mistakes in"} +{"text": "In the original article, there was a mistake in"} +{"text": "After 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incorrect versions of"} +{"text": "Following publication, the authors realized that"} +{"text": "In the original publication of the article , the fig"} diff --git a/PMC_clustering_98.jsonl b/PMC_clustering_98.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..360016939cfc7aed1ed6700d6937a41a848b4406 --- /dev/null +++ b/PMC_clustering_98.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:7951b4b147bd88ce8a5299f580c3bf64f419a283b4cdfff8bd59a02803c64ebe +size 61526567 diff --git a/PMC_clustering_99.jsonl b/PMC_clustering_99.jsonl new file mode 100644 index 0000000000000000000000000000000000000000..45f80c9a55d058dbba4b3bc081f2d2fb5a8750a9 --- /dev/null +++ b/PMC_clustering_99.jsonl @@ -0,0 +1,3 @@ +version https://git-lfs.github.com/spec/v1 +oid sha256:248ced0b71b6c69eb73ab1d9fec30faac7fdaf74e85d380e3c1f4d81454e4363 +size 87408643